A Phase 1 Study to Demonstrate the Relative Bioavailability of Fixed Dose Combinations of Ambrisentan and Tadalafil in Healthy Subjects
Phase: PHASE1
Conditions: Hypertension, Pulmonary
Sponsor: GlaxoSmithKline

2015N232335_03
CONFIDENTIAL
The GlaxoSmithKline group of companies
201964
1
TITLE PAGE
Division: Worldwide Development
Information Type: Protocol Amendment
Title:
A Phase 1 study to demonstrate the relative bioavailability and 
bioequivalence of fixed dose combinations of ambrisentan and 
tadalafil in healthy subjects
Compound Number:
GSK3380154, GSK1325760 and GF196960
Development Phase:
I
Effective Date:
25-MAY-2017
Protocol Amendment Number:  03
Author (s):
 
Revision Chronology
GlaxoSmithKline
Document Number
Date
Version
2015N232335_00
2015-DEC-16
Original
2015N232335_01
2016-APR-14
Amendment No. 1
Clarification of hypotension withdrawal criteria in Section 5.4.3, change to GSK medical 
monitor and minor administrative changes.
2015N232335_02
2017-FEB-08
Amendment No. 2

Inclusion of bioequivalence assessment in Part 3 to replace the optional food 
effect. 

Inclusion of additional dosage strength in Part 3. 

Change in Medical Monitor(s).
2015N232335_03
2017-MAY-25
Amendment No. 3
Inclusion of revised potential risk of clinical significance for tadalafil (Adcirca) following 
an update to the Summary of Product Characteristics and Patient Information Leaflet 
Change in Primary Medical Monitor
Changes to Data Analysis Considerations (section 9.3) and Pharmacokinetic Analyses 
9.4.1 to satisfy EMA guidelines.
Copyright 2017 the GlaxoSmithKline group of companies.  All rights reserved.  
Unauthorised copying or use of this information is prohibited.
 
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PPD
PPD

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INVESTIGATOR PROTOCOL AGREEMENT PAGE
For protocol number 201964
I confirm agreement to conduct the study in compliance with the protocol, as amended by 
this protocol amendment.
I acknowledge that I am responsible for overall study conduct. I agree to personally 
conduct or supervise the described study.
I agree to ensure that all associates, colleagues and employees assisting in the conduct of 
the study are informed about their obligations. Mechanisms are in place to ensure that site 
staff receives the appropriate information throughout the study. 
Investigator Name:
Investigator Address:
Investigator Phone Number:
Investigator Signature
Date
 
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TABLE OF CONTENTS
PAGE
1.
PROTOCOL SYNOPSIS FOR STUDY 201964........................................................7
2.
INTRODUCTION....................................................................................................11
2.1.
Study Rationale ..........................................................................................11
2.2.
Brief Background ........................................................................................12
3.
OBJECTIVE(S) AND ENDPOINT(S) ......................................................................14
4.
STUDY DESIGN ....................................................................................................14
4.1.
Overall Design ............................................................................................14
4.2.
Treatment and Duration..............................................................................16
4.3.
Type and Number of Subjects.....................................................................21
4.4.
Design Justification.....................................................................................21
4.5.
Dose Justification........................................................................................21
4.6.
Benefit: Risk Assessment ...........................................................................22
4.6.1.
Risk Assessment .........................................................................22
4.6.2.
Benefit Assessment.....................................................................24
4.6.3.
Overall Benefit: Risk Conclusion..................................................24
5.
SELECTION OF STUDY POPULATION AND WITHDRAWAL CRITERIA .............25
5.1.
Inclusion Criteria.........................................................................................25
5.2.
Exclusion Criteria........................................................................................26
5.3.
Screening/Baseline/Run-in Failures............................................................28
5.4.
Withdrawal/Stopping Criteria.......................................................................28
5.4.1.
Liver Chemistry Stopping Criteria ................................................28
5.4.1.1.
Study Treatment Restart or Rechallenge....................29
5.4.2.
QTc Stopping Criteria ..................................................................29
5.4.3.
Hypotension.................................................................................30
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria .........................30
5.5.
Subject and Study Completion....................................................................30
6.
STUDY TREATMENT ............................................................................................30
6.1.
Investigational Product and Other Study Treatment....................................30
6.1.1.
Retention Samples ......................................................................36
6.2.
Medical Devices..........................................................................................36
6.3.
Treatment Assignment................................................................................36
6.4.
Blinding.......................................................................................................38
6.5.
Packaging and Labeling..............................................................................38
6.6.
Preparation/Handling/Storage/Accountability..............................................38
6.7.
Compliance with Study Treatment Administration.......................................38
6.8.
Treatment of Study Treatment Overdose....................................................38
6.9.
Treatment after the End of the Study ..........................................................39
6.10.
Lifestyle and/or Dietary Restrictions............................................................39
6.10.1.
Meals and Dietary Restrictions ....................................................39
6.10.2.
Caffeine, Alcohol, and Tobacco ...................................................40
6.10.3.
Activity .........................................................................................40
6.11.
Concomitant Medications and Non-Drug Therapies....................................40
6.11.1.
Permitted Medications and Non-Drug Therapies..........................40
 
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6.11.2.
Prohibited Medications and Non-Drug Therapies.........................40
7.
STUDY ASSESSMENTS AND PROCEDURES .....................................................40
7.1.
Time and Events Table...............................................................................42
7.2.
Screening and Critical Baseline Assessments ............................................43
7.2.1.
Demographic/Medical History Assessments ................................43
7.3.
Pharmacokinetics .......................................................................................43
7.3.1.
Blood Sample Collection..............................................................43
7.3.2.
Sample Analysis ..........................................................................43
7.4.
Safety .........................................................................................................44
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)..........44
7.4.1.1.
Time period and Frequency for collecting AE 
and SAE information...................................................44
7.4.1.2.
Method of Detecting AEs and SAEs ...........................45
7.4.1.3.
Follow-up of AEs and SAEs........................................45
7.4.1.4.
Regulatory Reporting Requirements for SAEs............45
7.4.2.
Physical Exams ...........................................................................45
7.4.3.
Vital Signs....................................................................................46
7.4.4.
Electrocardiogram (ECG).............................................................46
7.4.5.
Clinical Safety Laboratory Assessments ......................................46
8.
DATA MANAGEMENT ...........................................................................................48
9.
STATISTICAL CONSIDERATIONS AND DATA ANALYSES .................................48
9.1.
Hypotheses.................................................................................................48
9.2.
Sample Size Considerations.......................................................................49
9.2.1.
Sample Size Assumptions ...........................................................49
9.2.2.
Sample Size Sensitivity................................................................50
9.2.3.
Sample Size Re-estimation or Adjustment...................................51
9.3.
Data Analysis Considerations .....................................................................51
9.3.1.
Analysis Populations....................................................................52
9.3.1.1.
Safety Population .......................................................52
9.3.1.2.
Pharmacokinetic Concentration Population ................52
9.3.1.3.
Pharmacokinetic Parameter Population......................52
9.3.2.
Interim Analysis ...........................................................................52
9.4.
Key Elements of Analysis Plan ...................................................................52
9.4.1.
Pharmacokinetic Analyses...........................................................52
9.4.2.
Safety Analyses...........................................................................53
10. STUDY GOVERNANCE CONSIDERATIONS........................................................54
10.1.
Posting of Information on Publicly Available Clinical Trial Registers............54
10.2.
Regulatory and Ethical Considerations, Including the Informed 
Consent Process ........................................................................................54
10.3.
Quality Control (Study Monitoring) ..............................................................55
10.4.
Quality Assurance.......................................................................................55
10.5.
Study and Site Closure ...............................................................................55
10.6.
Records Retention......................................................................................56
10.7.
Provision of Study Results to Investigators, Posting of Information 
on Publically Available Clinical Trials Registers and Publication .................56
11. REFERENCES.......................................................................................................58
 
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12. APPENDICES........................................................................................................60
12.1.
Appendix 1: Abbreviations and Trademarks................................................60
12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments ..............................................................................................61
12.3.
Appendix 3: Definition of and Procedures for Recording, Evaluating, 
Follow-Up and Reporting of Adverse Events...............................................63
12.3.1.
Definition of Adverse Events........................................................63
12.3.2.
Definition of Serious Adverse Events...........................................64
12.3.3.
Definition of Cardiovascular Events .............................................65
12.3.4.
Recording of AEs and SAEs ........................................................66
12.3.5.
Evaluating AEs and SAEs............................................................66
12.3.6.
Reporting of SAEs to GSK...........................................................68
12.4.
Appendix 4 High Fat Meal Content .............................................................69
12.5.
Appendix 5: Protocol Changes....................................................................70
12.5.1.
Protocol changes for Amendment 01 (14-Apr-2016) from 
Original Protocol (16-Dec-2015) ..................................................70
12.5.2.
Protocol changes for Amendment 2 (08-FEB-2017) from 
Amendment 1(14-Apr-2016) ........................................................72
12.5.3.
Protocol changes for Amendment 3 (25-MAY-2017) from 
Amendment 3 (8-Feb-2017).......................................................122
 
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1.
PROTOCOL SYNOPSIS FOR STUDY 201964
Rationale
This study is designed initially to compare the relative bioavailability of a number of 
fixed dose combinations (FDCs) of ambrisentan and tadalafil (Part 1 and 2) and 
consequently the bioequivalence of the FDC of different dose strength (Part 3A and 3B). 
Dependent on formulation work, the study will allow up to 9 new FDCs to be compared 
with the reference of ambrisentan and tadalafil monotherapies taken concurrently. 
Objective(s)/Endpoint(s)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following administration 
of candidate tablet formulations of the FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), and 
AUC(0-t) of ambrisentan and tadalafil in FDC 
and reference treatments
To establish bioequivalence of the candidate FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fed and fasted conditions.
AUC(0-t),AUC(0-inf) ,Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC (0-t), AUC(0-inf) ,Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 20 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 20 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC (0-t), AUC(0-inf) ,Cmax
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fed and fasted conditions.
Plasma PK parameters including; AUC(0-
t)/AUC(0-inf) ,tmax, t1/2 of ambrisentan and 
tadalafil in FDC and reference treatments
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fed and fasting
conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
 
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Overall Design
This is a single centre, Phase 1, single dose, randomised, open label crossover study in 
healthy volunteers with 3 study parts. Part 1 will include a 5 way cross-over. Part 2 and 
Part 3 (A&B) will each include a 4 way cross-over. All subjects will attend the unit for 
Screening within 31 days of their first dose. Eligible subjects will be randomised to order 
of treatment in that study parts in which they are included and the order in which these 
treatments are administered will be in accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in Parts 1 and 2. In Part 3 
the washout will be at least 10 days.  A Follow-Up Visit should be completed within 7-14 
days of completion of a subject’s last dose.  The utility of each of the 3 study parts is as 
follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part will include an evaluation of the bioavailability, safety and tolerability of 
3 granulation forms of a single FDC (ambrisentan 10mg + tadalafil 40 mg) identified 
from Part 1. These data will be compared to that for the reference of the 2 monotherapy 
components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All doses are 
taken in the fasted state.
Part 3 A
Part 3A of the study is set to establish bioequivalence between the candidate FDC from 
Part 2. This study part will have 4 dose sessions and will assess the bioequivalence, in 
both the fed and fasted state, of the FDC against the reference ambrisentan 10 mg + 
tadalafil 40mg monotherapies in the fed and fasted state.  The fed arms of this part will 
have a standard high fat breakfast. (EMEA, 2010).
Part 3 B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 40mg and 
ambrisentan 5 mg + tadalafil 20mg). This study part will have 4 dose sessions and will
assess the bioequivalence of the two FDC dose strengths against the reference 
 
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ambrisentan 5 mg + tadalafil 40mg and ambrisentan 5 mg + tadalafil 20mg 
monotherapies in the fasted state.
Treatment Arms and Duration
The proposed treatment arms for each study part are described here.
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)
Part 3A (fed and fasted)
Part 3B 
(fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 1
(10mg/40mg)
ambrisentan and tadalafil 
FDC ) (10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 2
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg) fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 3
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
 
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Type and Number of Subjects
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable subjects 
complete each part of the study.  Approximately 33 healthy subjects will be randomized
to study Part 3A, such that at least 26 evaluable subjects complete part 3A of the study. 
An additional 33 healthy subejcts will be randomised to Part 3B, such that at least 26 
evaluable subjects complete part 3B of the study.
Analysis
No formal hypothesis will be tested for Part 1 and Part 2 of the study. An estimation 
approach will be used to estimate the bioavailability of the FDC formulations relative to 
the monotherapies taken concurrently.  
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 10 
mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 10 mg & 
tadalafil 40 mg taken concurrently (reference) in healthy subjects in both the fed and 
fasted states. The null hypothesis is that the true ratio of the geometric mean of the test 
treatment to the geometric mean of the reference treatment, (test)/(reference), for each 
primary PK endpoint, is either less than 0.80 or greater than 1.25.  The alternate 
hypothesis is that the true ratio of the test treatment geometric mean to the reference 
treatment geometric mean is at least 0.80 and no greater than 1.25.  Symbolically, this is 
expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e, treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test (TOST) 
procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be used to test 
this set of hypotheses. This is equivalent to requiring that a 90% interval for the true ratio 
of test to reference geometric means falls entirely within the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test the bioequivalence of a
FDC of ambrisentan 5 mg + tadalafil 40 mg (test) and of ambrisentan 5 mg + tadalafil 20 
mg (test) relative to reference monotherapies of  ambrisentan 5 mg & tadalafil 40 mg and 
of ambrisentan 5 mg & tadalafil 20 mg respectively taken concurrently (reference) in 
 
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healthy subjects in the fasted states. Therefore, the analysis approach for bioequivalence 
for Part 3A is  applicable to Part 3B.
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
2.
INTRODUCTION
Ambrisentan (E.U. trade name: Volibris), is an orally active endothelin receptor 
antagonist (ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first 
approved on 15 June 2007 in the US and on 21 April 2008 in the European Union (EU) 
and is currently approved in over 50 countries. In the EU, ambrisentan is indicated for 
treatment of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional 
Class (FC) II to III, including use in combination treatment (Volibris, 2015; Section 5.1). 
Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated with 
connective tissue disease.
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (EMEA, 2010; Schuirmann, 1987;Adcirca, 2015).
A recently completed study (Galiè, 2015a) has shown that patients with PAH who started 
initial combination therapy with ambrisentan and tadalafil had a significantly lower risk 
of clinical-failure events compared to those that started with ambrisentan or tadalafil 
monotherapy. Ambrisentan has recently received EU approval (20 November 2015) for 
use in combination treatment with tadalafil (Volibris, 2015, Section 5.1).
2.1.
Study Rationale
This study is designed initially to compare the relative bioavailability of a number of 
fixed dose combinations (FDCs) of ambrisentan and tadalafil (Part 1 and 2) and 
consequently the bioequivalence of the FDC of different dose strengths (Part 3A and 3B). 
Dependent on formulation work, the study will allow up to 9 new FDCs to be compared 
with the reference of ambrisentan and tadalafil monotherapies taken concurrently
The formulation(s) to be taken forward in Part 3A  will be primarily chosen based on the 
test/reference ratio for both AUC and Cmax for both components. Ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, tablet size, cost, ease of 
manufacture and stability would be considered. 
Part 3 will include evaluation of bioequivalence for 3 dose strengths of the FDC; 
10/40mg, 5/40mg and 5/20mg of ambrisentan and tadalafil.  
 
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2.2.
Brief Background
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris, 2015, Section 5.1). Efficacy has been shown in 
idiopathic PAH (IPAH) and in PAH associated with connective tissue disease (Volibris, 
2015). Ambrisentan is an oral, once daily, propanoic acid-based, ETA-selective 
Endothelin receptor antagonist (ERA) which targets the phospholipase-C-dependent 
endothelin pathway and which is known to play an essential role in mammalian 
cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (EMEA, 2010; Schuirmann, 
1987;Adcirca, 2015). Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) 
inhibitor which targets the NO pathway. Through inhibition of PDE-5, tadalafil increases 
cytoplasmic cGMP concentrations in the smooth muscle cells and enhances NO-mediated 
vasodilatation of the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy participants was performed to 
detect any significant PK interactions between tadalafil and ambrisentan when co-
administered (Spence, 2009). From this study, it was concluded that there is no clinically 
significant PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when 
combined. Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
 
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administered.  There were no SAE’s in the study. Three participants withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one participantbecause of myalgia 
(severe), muscle fatigue and dizziness on tadalafil alone and one because of headache 
(severe) on the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild 
and is a listed event for ambrisentan. There were a total of 7 participants with mild 
tachycardia. There were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. 
There were 3 events on the first day of tadalafil 40 mg given 3 days after ambrisentan 10 
mg. There was one event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of 
ambrisentan 10 mg. Taken together these data suggest that in healthy subjects a mild 
tachycardia may result from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, 2015, (EMEA, 
2010; Schuirmann, 1987;Adcirca, 2015).
The AMBITION clinical study (Galiè, 2015a and GSK document number 
2014N193963_00), which evaluated the time to first clinical failure event, a composite 
endpoint, shows a robust clinical benefit (50% hazard reduction) for PAH patients 
initiated on a combination of ambrisentan and tadalafil when compared to PAH patients 
initiated on either medication as monotherapy.  The safety profile of the combination arm 
was consistent with the known safety data of the individual study drugs and no safety 
signals specific to combination treatment were identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 2008).
 
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3.
OBJECTIVE(S) AND ENDPOINT(S)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following administration 
of candidate tablet formulations of the FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), and 
AUC(0-t) of ambrisentan and tadalafil in FDC 
and reference treatments
To establish bioequivalence of the candidate FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fed and fasted conditions.
AUC(0-t), AUC(0-inf), Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC(0-t), AUC (0-inf), Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 20 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 20 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC (0-t), AUC (0-inf) ,Cmax
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting and fed conditions.
Plasma PK parameters including; AUC(0-
t)/AUC(0-inf), tmax, t1/2 of ambrisentan and 
tadalafil in FDC and reference treatments
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fed and fasting 
conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
4.
STUDY DESIGN
4.1.
Overall Design
This is a single centre, Phase 1, single dose, randomised, open label, crossover study in 
healthy volunteers with 3 study parts; Part 1 will include a 5 way cross-over, and Part 2 
and 3 (A&B) will each include a 4 way cross-over. See Figure 1 for study schematic.
 
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Figure 1
Study Schematic
Screening
≤ 31 days
Follow 
Up
Screening 
≤ 31 days from
Subjects 
Randomised to  
4 dose sessions 
(3 granulation 
formulations + 
reference)
Follow 
Up
Subjects 
randomised to 4 
dose sessions 
(FDC Fed & 
fasted + 
reference (fed 
&fasted))
Screening 
≤ 31 days
Follow 
Up
~4 Weeks
~4 Weeks
~4 Weeks
PART 1 (BA) n=20
PART 2 (BA) n=20
PART 3A (BE 10/40) n=33
Subjects 
Randomised to 
up to 5 dose 
sessions (4 FDC 
formulations 
and the 
reference)
Screening 
≤ 31 days
Subjects 
Randomised to 
4 dose sessions    
(2 FDC 
formulations 
(fasted) + 2 
reference fasted)
Follow 
Up
PART 3B (BE 5/40 & 5/20)
n=33
~4 Weeks
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in study Part 1 and 2. In 
study Part 3 there will be a minimum of 10 days between each dose.  A Follow-Up Visit 
should be completed within 7-14 days of completion of a subject’s last dose. 
The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have 5 dose sessions and will be used  to characterise the relative 
bioavailability, safety and tolerability of   four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
 
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Part 2
This study part will have 4 dose sessions and will evaluate the bioavailability, safety and 
tolerability of  3 different granulation sizes for a single FDC (ambrisentan 10 mg + 
tadalafil 40 mg) compared to the reference of the 2 monotherapy components taken 
concurrently (ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted 
state.
Part 3A
Part 3A of the study is set to establish bioequivalence between the candidate FDC from 
Part 2. This study part will have 4 dose sessions and will assess the bioequivalence, in 
both the fed and fasted state, of the FDC against the reference  ambrisentan 10 mg + 
tadalafil 40mg monotherapies in the fed and fasted state. The fed arms of this part will 
have a standard high fat breakfast (EMEA, 2010).
Part 3B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 40mg and 
ambrisentan 5 mg + tadalafil 20mg) . This study part will have 4 dose sessions and will
assess the bioequivalence of the two FDC dose strengths against the reference 
ambrisentan 5 mg + tadalafil 40mg monotherapies and ambrisentan 5 mg + tadalafil 
20mg monotherapies in the fasted state.
4.2.
Treatment and Duration
The treatments for each study part of the study are listed in Table 1. All treatments are 
single dose. Subjects will be randomised to the order of treatments in the parts of the 
study they are included in.
The study has 3 parts and ongoing analysis of pharmacokinetic data will be used to 
determine the formulations produced and tested in subsequent parts.
Part 1
Part 1 of the study will be utilised to evaluate 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2
and 3. 
 
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Following completion of Part 1 there will be a pause prior to Part 2, so that different 
granulation forms of a single FDC can be produced and data included and approved in 
any required update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for 3 granulation size forms of a single FDC 
formulation selected from Part 1. These will be based on a single successful formulation 
identified in Part 1 reformulated with the final active pharmaceutical ingredient (API).
Success will be defined with the same criteria as those in Part 1.
Part 3 (A and B)
The BE part of the study will investigate the bioequivalence of the FDC ambrisentan 10 
mg + tadalafil 40 mg in part 3A and the bioequivalence of the FDC ambrisentan 5 mg + 
tadalafil 40 mg and ambrisentan 5 mg + tadalafil 20 mg in Part 3B.  
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted) 
Part 3A (fed and fasted)
Part 3B (fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 1
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and tadalafil 
FDCgranulation 2
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg) 
fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 3
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg), fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg), fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
The duration for a subject on this study is described in Figure 2. This figure assumes all 5 
dose sessions are used for Part 1. However, this is an approximation as minimum of 7 
days is expected between doses in Parts 1 and 2 and a minimum of 10 days in Part 3.
 
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Figure 2
Study Duration
Part 1 can last up to 73 days.
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
Dose 
Period 5 
start 
7 
days
7 
days
7 
days
7 
days
7 to 
14 
days
Follow-up
 
 
 
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Part 2  will last approximately 9 weeks
Part 3A and B will last approximately 11 weeks
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
10 
days
10  
days
10 
days
7 to 14  
days
Follow-up
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
7 days
7 days
7 days
7 to 14  
days
Follow-up
 
 
 
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A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
4.3.
Type and Number of Subjects
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable subjects 
complete each part of the study.  Approximately 33 healthy subjects will be randomized
to study Part 3A, such that at least 26 evaluable subjects complete part 3A of the study. 
An additional 33 healthy subjects will be randomised to Part 3B, such that at least 26 
evaluable subjects complete part 3B of the study.
The estimation of the number of subjects for each study part is detailed further in Section 
9.2 Sample Size Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
4.4.
Design Justification
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, nor is there any need to blind study treatment. The inclusion of the two
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study contains three parts: Parts 1 and 2 will provide PK data for different candidate 
FDC formulations and will enable the assessment of bioequivalence of the FDC versus 
the combination of the individual reference products in Part 3. The interdependency of 
the 3 study parts is described in detail in Section 4.2. 
4.5.
Dose Justification
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of 40mg for tadalafil and 10mg for ambrisentan, are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
 
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be tolerated in healthy subjects. Within Part 3 of the study all dosage forms of the FDC 
that are proposed for marketing authorisation will be tested for bioequivalence versus the 
appropriate doses of reference products.  These doses are required in clinical practice to 
allow safe up- and down-titration of the FDC in patients with PAH.  
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimumof 10 days, between doses, 
is also sufficient for clearance of the previous dose during the assessmet of 
bioequivalence.
4.6.
Benefit: Risk Assessment
There is no direct benefit for healthy subjects participating in this study.  The risks to the 
healthy subjects based upon previous experience indicate no expectation of SAE and mild 
to moderate AE.  The benefit/risk remains satisfactory for this healthy subject study.
Summaries of findings from clinical studies conducted with both the Investigational 
Product taken as monotherapies and in combination can be found in: 

Ambrisentan IB (Volibris IB, 2015)

Volibris (Ambrisentan) EMA SmPC (Volibris, 2015) 

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010; Schuirmann, 1987;Adcirca, 2017)
As described in Section 2.2, mild tachycardia was the most common (7/26, 27%) adverse 
event seen in healthy subjects following combination dosing with 10mg ambrisentan and 
40mg tadalafil (Spence, 2009). There were three subjects (3/26, 11.5%) withdrawn due to 
AEs (e.g., anaemia [mild]; myalgia [severe], muscle fatigue, and dizziness; and headache 
[severe]).    No SAEs or additional risks were observed with combination use in healthy 
subjects.
The following section outlines the risk assessment and mitigation strategy for this 
protocol:  
4.6.1.
Risk Assessment
Risks of clinical significance, identified in subjects with PAH and the mitigation strategy 
identified for this study in healthy subjects are captured in Table 2. 
 
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Table 2
Risks of Clinical Significance and Mitigation Strategy
Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
Investigational Product: GSK1325760 (ambrisentan-Volibris)

Teratogenicity

Anaemia (decreased 
haemoglobin, decreased 
haematocrit)

Hypersensitivity reactions 
(e.g. angioedema, rash, 
pruritus)

Headache (including sinus 
headache, migraine)

Dizziness

Cardiac failure

Palpitation

Hypotension

Flushing

Epistaxis

Dyspnoea

Upper respiratory  
congestion, sinusitis, 
nasopharyngitis, rhinitis

Abdominal pain

Constipation

Nausea, vomiting, 
diarrhoea

Hepatic transaminases 
increased

Peripheral oedema, fluid 
retention

Chest pain/discomfort

Asthenia and fatigue
Ambrisentan IB (Volibris IB, 
2015)
Volibris (Ambrisentan) EMA 
SmPC (Volibris, 2015): 
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision. 

Exclusion of women of 
childbearing potential 
(teratogenicity)

Laboratory assessments 
per protocol

Physical assessment per 
protocol

Routine vital signs per 
protocol

Subjects remain in the 
clinical unit, under medical 
supervision, for all doses 
and until completion of 
safety assessments at 
48 hrs post dose.

Single doses used in the 
study
Investigational Product: GF196960 (tadalafil - Adcirca)

Hypersensitivity reactions

Headache

Syncope,

Migraine

Blurred vision

Sudden decrease or loss 
of vision (Non-Arteric 
Anterior Ischemic Optic 
Neuropathy (NAION))

Palpitations
Adcirca (Tadalafil) EMA SmPC 
(EMEA, 2010; Schuirmann, 
1987;Adcirca, 2017)
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision.

Routine vital signs per 
protocol

Physical assessment per 
protocol

Subjects remain in the 
clinical unit for all doses, 
under medical supervision 
and until completion of 
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy

Nasopharyngitis (including 
nasal congestion, sinus 
congestion and rhinitis)

Epistaxis

Nausea,

Dyspepsia (including 
abdominal pain/discomfort)

Vomiting,

Gastroesophageal reflux

Rash

Myalgia,

Back pain

Pain in extremity (including 
limb discomfort)

Increased uterine bleeding

Facial oedema,

Chest pain

Sudden decrease or loss 
of hearing
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GSK3380154 (ambrisentan-tadalafil-FDC)
The risks for the combination 
are the same as the 2 
monotherapies; no extra risks 
have been identified for the 
combination.
As for GSK1325760 and 
GF196960 above
4.6.2.
Benefit Assessment
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation and to 
demonstrate bioequivalence versus reference products for a fixed dose combination of 
ambrisentan plus tadalafil to use as first line therapy in PAH patients.
4.6.3.
Overall Benefit: Risk Conclusion
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 2. In 
addition review of safety information from Part 1 of the study is consistent with this 
position. So, the benefit:risk is appropriate for this study and to enable development of 
this investigational product. The overall benefit: risk therefore remains positive.
 
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5.
SELECTION OF STUDY POPULATION AND 
WITHDRAWAL CRITERIA
Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in:

Volibris (Ambrisentan) EMA SmPC (Volibris, 2015)

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010; Schuirmann, 1987;Adcirca, 2017)
Deviations from inclusion and exclusion criteria are not allowed because they can 
potentially jeopardize the scientific integrity of the study, regulatory acceptability or 
subject safety. Therefore, adherence to the criteria as specified in the protocol is essential.
5.1.
Inclusion Criteria
A subject will be eligible for inclusion in this study only if all of the following criteria 
apply:
AGE
1. Between 18 and 60 years of age inclusive, at the time of signing the informed 
consent.
TYPE OF SUBJECT AND DIAGNOSIS INCLUDING DISEASE SEVERITY
2. Healthy as determined by a responsible and experienced physician, based on a 
medical evaluation including medical history, physical examination, laboratory tests, 
vital signs and cardiac monitoring (ECG and 24 hour Holter).  A subject with a 
clinical abnormality or laboratory parameter(s) which is/are not specifically listed in 
the inclusion or exclusion criteria, outside the reference range for the population 
being studied may be included only if the Investigator, in consultation with the GSK 
Medical Monitor if required, judges and documents that the finding is unlikely to 
introduce additional risk factors and will not interfere with the study procedures. 
WEIGHT
3. Body weight ≥ 50 kg (110 lbs) for men and ≥ 45 kg (99lbs) for women and body 
mass index (BMI) within the range 18– 30 kg/m2 (inclusive)
SEX
4. Male or Female
Females must be the following:
Non-reproductive potential defined as:
 
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
Pre-menopausal females with one of the following:
Documented tubal ligation
Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion 
Hysterectomy
Documented Bilateral Oophorectomy

Documented Postmenopausal defined as 12 months of spontaneous amenorrhea
INFORMED CONSENT
5. Capable of giving signed informed consent as described in Section 10 which includes 
compliance with the requirements and restrictions listed in the consent form and in
this protocol.
5.2.
Exclusion Criteria
A subject will not be eligible for inclusion in this study if any of the following criteria 
apply:
CONCURRENT CONDITIONS/MEDICAL HISTORY (INCLUDES LIVER 
FUNCTION AND QTC INTERVAL)
1.
A blood pressure <100/55 mm Hg.
2.
Haemoglobin below normal range:

Hb < 133 g/L for males 

Hb< 114 g/L for females
3.
ALT and bilirubin >1.5xULN (isolated bilirubin >1.5xULN is acceptable if bilirubin 
is fractionated and direct bilirubin <35%). 
4.
Current or chronic history of liver disease, or known hepatic or biliary abnormalities 
(with the exception of Gilbert's syndrome or asymptomatic gallstones)
5.
QTc> 450 msec
NOTES:

The QTc is the QT interval corrected for heart rate according to Bazett’s 
formula (QTcB), Fridericia’s formula (QTcF), and/or another method, machine-
read or manually over-read.

The specific formula that will be used to determine eligibility and 
discontinuation for an individual subject should be determined prior to initiation 
of the study. In other words, several different formulae cannot be used to 
calculate the QTc for an individual subject and then the lowest QTc value used 
to include or discontinue the subject from the trial.

For purposes of data analysis, QTcB, QTcF, another QT correction formula, or a 
composite of available values of QTc will be used as specified in the Reporting 
 
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and Analysis Plan (RAP).
CONCOMITANT MEDICATIONS
6.
Use of prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements (including St John’s Wort) within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose 
of study medication, unless in the opinion of the Investigator and GSK Medical 
Monitor the medication will not interfere with the study procedures or compromise 
subject safety. 
RELEVANT HABITS
7.
History of regular alcohol consumption within 6 months of the study defined as: 

An average weekly intake of >21 units for males or >14 units for females.  One 
unit is equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 
ml) of wine or 1 (25 ml) measure of spirits.
8.
Smoking more than 5 cigarettes per week and subjects must be able to abstain from 
smoking for a 24 hour period prior to dose and any time whilst in the clinical unit.
CONTRAINDICATIONS
9.
History of sensitivity to any of the study medications, or components thereof or a 
history of drug or other allergy that, in the opinion of the Investigator or Medical 
Monitor, contraindicates their participation.
DIAGNOSTIC ASSESSMENTS AND OTHER CRITERIA
10. Presence of hepatitis B surface antigen (HBsAg), positive hepatitis C antibody test 
result at Screening or within 3 months prior to first dose of study treatment. . 
11. A positive test for HIV antibody.
12. A positive pre-study drug/alcohol screen.  
13. Where participation in the study would result in donation of blood or blood products 
in excess of 500 mL within previous 3 months
14. The subject has participated in a clinical trial and has received an investigational 
product within the following time period prior to the first dosing day in the current 
study: 3 months, 5 half-lives or twice the duration of the biological effect of the 
investigational product (whichever is longer).
15. Exposure to more than four new chemical entities within 12 months prior to the first 
 
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dosing day.
5.3.
Screening/Baseline/Run-in Failures
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT) 
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.4).
5.4.
Withdrawal/Stopping Criteria
The following actions must be taken in relation to a subject who fails to attend the clinic 
for a required study visit:

The site must attempt to contact the subject and re-schedule the missed visit as 
soon as possible. 

The site must counsel the subject on the importance of maintaining the assigned 
visit schedule and ascertain whether or not the subject wishes to and/or should 
continue in the study. 

In cases where the subject is deemed ‘lost to follow up’, the investigator or 
designee must make every effort to regain contact with the subject (where 
possible, 3 telephone calls and if necessary a certified letter to the subject’s last 
known mailing address or local equivalent methods). These contact attempts 
should be documented in the subject’s medical record. 

Should the subject continue to be unreachable, only then will he/she be considered 
to have withdrawn from the study with a primary reason of “Lost to Follow-Up”. 
A subject may withdraw from study treatment at any time at his/her own request, or may 
be withdrawn at any time at the discretion of the investigator for safety, behavioural or 
administrative reasons. If a subject withdraws from the study, he/she may request 
destruction of any samples taken, and the investigator must document this in the site 
study records. 
5.4.1.
Liver Chemistry Stopping Criteria
Liver chemistry stopping and increased monitoring criteria have been designed to 
assure subject safety and evaluate liver event etiology (in alignment with the FDA 
premarketing clinical liver safety guidance). 
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf
 
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Study treatment will be discontinued for a subject if liver chemistry stopping criteria are 
met:
Phase I Liver Chemistry Stopping Criteria – Liver Stopping Event Algorithm
Continue Study Treatment 
Discontinue Study Treatment 
*INR value not applicable to subjects on anticoagulants 
ALT ≥3xULN 
No
Yes
Must refer to Liver Safety Required Actions and Follow up Assessments  section in the Appendix
Report as an SAE if possible Hy’s Law case: ALT≥3xULN and  Bilirubin≥2xULN (>35% direct)  or 
INR>1.5, if measured*
Liver Safety Required Actions and Follow-Up Assessments Section can be found in 
Appendix 2: Liver Safety Required Actions and Follow-Up Assessments
5.4.1.1.
Study Treatment Restart or Rechallenge
Study treatment restart or re-challenge after liver chemistry stopping criteria are met by 
any subject participating in this study is not allowed.
5.4.2.
QTc Stopping Criteria

The same QT correction formula must be used for each individual subject to 
determine eligibility for and discontinuation from the study.  This formula may 
not be changed or substituted once the subject has been enrolled.

For example, if a subject is eligible for the protocol based on QTcF, then QTcF 
must be used for discontinuation of this individual subject as well.

Once the QT correction formula has been chosen for a subject’s eligibility, the 
same formula must continue to be used for that subject for all QTc data being 
collected for data analysis.  Safety ECGs and other non-protocol specified 
ECGs are an exception.  

The QTc should be based on averaged QTc values of triplicate electrocardiograms 
obtained over a brief (e.g., 5-10 minute) recording period. 
A subject that meets either bulleted criterion below will be withdrawn from the study.  

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
Change from baseline: QTc >60 msec 
5.4.3.
Hypotension
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 mmHG and diastolic <50 mm HG confirmed by 
triplicate reading taken up to 5 minutes apart and is judged clinically significant 
and symptomatic by the investigator.
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria
For an individual study participant, stopping criteria include, but are not limited to:
Adverse events, or significant changes in any of the safety assessments, that put the 
safety of the individual at risk (e.g., ECG, vital signs, laboratory tests, etc), as judged by 
the Principal Investigator in consultation with the Medical Monitor if necessary.
5.5.
Subject and Study Completion
A completed subject is one who has completed all dosing periods of the study including 
the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
6.
STUDY TREATMENT
6.1.
Investigational Product and Other Study Treatment
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Parts 1, 2 and 3 of the study are shown in Table 3, Table 
4 and Table 5. 
 
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Table 3
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 90 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.45
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Address of 
reference 
manufacturer
NA
NA
NA
NA
Glaxo Operations UK 
Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 4
Study Treatment for Part 2
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-tadalafil-
FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 granulation 1
(10mg/40mg)
FDC-G2 granulation 2
(10mg/40mg)
FDC-G3 granulation 3 
(10mg/40mg)
Each tablet contains 10 mg 
of ambrisentan, 
approximately 90 mg of 
lactose (as monohydrate), 
approximately 0.25 mg of 
lecithin (soya) (E322) and 
approximately 0.45 mg of 
Allura red AC Aluminium 
Lake 
Each film-coated tablet 
contains 20 mg 
tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage level(s):
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 40 
mg tadalafil and 10 mg 
of ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 10 mg 
of ambrisentan.
Each tablet contains 20 
mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations UK Ltd 
(trading as GlaxoWellcome 
Operations), Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la Industria 
30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 5
Study Treatment for Part 3
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-tadalafil-
FDC)
GSK1325760 
(ambrisentan-Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 or 2 or3
(10mg/40mg)
FDC  (5mg/40mg)
FDC  (5mg/20mg)
Each tablet contains 
10 mg of ambrisentan, 
approximately 90 mg 
of lactose (as 
monohydrate), 
approximately 0.25 
mg of lecithin (soya) 
(E322) and 
approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each tablet 
contains 5 mg of 
ambrisentan, 
approximately 95
mg of lactose (as 
monohydrate), 
approximately 0.25 
mg of lecithin 
(soya) (E322) and 
approximately 0.11 
mg of Allura Red
AC Aluminium 
Lake
Each film-coated 
tablet contains 20 
mg tadalafil, 245
mg lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 5 mg 
of ambrisentan.
Each tablet contains 
20 mg tadalafil and 5
mg of ambrisentan.
Each tablet contains 
10 mg of ambrisentan.
Each tablet 
contains 5 mg of 
ambrisentan.
Each tablet 
contains 20 mg 
tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-tadalafil-
FDC)
GSK1325760 
(ambrisentan-Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations UK 
Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Glaxo Operations 
UK Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 Alcobendas
Madrid
Spain
 
 
 
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6.1.1.
Retention Samples
GSK will send five times (5x) the drug product necessary for pivotal bioequivalency 
evaluation in Part 3 of the study for analytical testing for all tablet strengths used in the 
study. The retain samples are in addition to the supply of drug product sufficient to 
complete the study. Retention samples will be sent to the clinical CRO in the same bulk 
container. To ensure that reserve samples are representative of the same batches provided 
for the clinical study the clinical site will randomly select the supplies for the study from 
the supply received. The clinical site must retain enough reserve samples to permit FDA 
to perform five times all of the release tests required in the application.
6.2.
Medical Devices
No GSK manufactured devices (or devices manufactured for GSK by a third party) are 
provided for use in this study.
6.3.
Treatment Assignment
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software.
The treatment key for Parts 1, 2 and 3 are described in Table 6.
Table 6
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 2
FG1
ambrisentan and tadalafil FDC-G1 (10mg/40mg) granulation size 1
FG2
ambrisentan and tadalafil FDC-G2 (10mg/40mg) granulation size 2
FG3
ambrisentan and tadalafil FDC-G3 (10mg/40mg) granulation size 3
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3A
X1
ambrisentan and tadalafil FDC-G1 or 2 or 3 (10mg/40mg) fed
X2
ambrisentan and tadalafil FDC G1 or 2 or 3 (10mg/40mg) fasted
 
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Treatment
Description
R1
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fed
R2
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fasted
Part 3B
Y1
ambrisentan and tadalafil FDC (5mg/40mg), fasted
Y2
ambrisentan and tadalafil FDC (5mg/20mg), fasted
R3
ambrisentan and tadalafil monotherapies taken concurrently (5mg/40mg), 
fasted
R4
ambrisentan and tadalafil monotherapies taken concurrently (5mg/20mg), 
fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 7. Additional treatment sequences may
be created based using the Latin Squares for Williams Designs if needed.
Table 7
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1/ F2/ R/ F3/ F4
F2/ F3/ F1/ F4/ R
F3/ F4/ F2/ R/ F1
F4/ R/ F3/ F1/ F2
R/ F1/ F4/ F2/ F3
F4/ F3/ R/ F2/ F1
R/ F4/ F1/ F3/ F2
F1/ R/ F2/ F4/ F3
F2/ F1/ F3/ R/ F4
F3/ F2/ F4/ F1/ R
1:1:1:1:1:1:1:1:1:1
Part 2
4
FG1/ FG2/ R/ FG3
FG2/ FG3/ FG1/ R
FG3/ R/ FG2/ FG1
R/ FG1/ FG3/ FG2
1:1:1:1
Part 3A
4
X1/ R1/ R2/ X2
R1/ X2/ X1/ R2
X2/ R2/ R1/ X1
R2/ X1/ X2/ R1
1:1:1:1
Part 3B
4
Y1/ R3/ R4/ Y2
R3/ Y2/ Y1/ R4
Y2/ R4/ R3/ Y1
R4/ Y1/ Y2/ R3
1:1:1:1
 
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6.4.
Blinding
This will be an open-label study. 
6.5.
Packaging and Labeling
The contents of the label will be in accordance with all applicable regulatory 
requirements.
6.6.
Preparation/Handling/Storage/Accountability
This will be detailed in a Study Specific Technical Agreement/Memo (TTS) which will 
be accompanied by a Quality Agreement.

Only subjects enrolled in the study may receive study treatment and only 
authorized site staff may supply or administer study treatment.  All study 
treatments must be stored in a secure environmentally controlled and monitored 
(manual or automated) area in accordance with the labelled storage conditions 
with access limited to the investigator and authorized site staff. 

The investigator, institution, or the head of the medical institution (where 
applicable) is responsible for study treatment accountability, reconciliation, and 
record maintenance (i.e. receipt, reconciliation and final disposition records).  

Further guidance and information for final disposition of unused study treatment 
are provided in the SRM. 

Under normal conditions of handling and administration, study treatment is not 
expected to pose significant safety risks to site staff.  

A Material Safety Data Sheet (MSDS)/equivalent document describing 
occupational hazards and recommended handling precautions either will be 
provided to the investigator, where this is required by local laws, or is available 
upon request from GSK.
6.7.
Compliance with Study Treatment Administration
When subjects are dosed at the site, they will receive study treatment directly from the 
investigator or designee, under medical supervision.  The date and time of each dose 
administered in the clinic will be recorded in the source documents.  The dose of study 
treatment and study subject identification will be confirmed at the time of dosing by a 
member of the study site staff other than the person administering the study treatment.  
Study site personnel will examine each subject’s mouth to ensure that the study treatment 
was ingested.
6.8.
Treatment of Study Treatment Overdose
For this study, any dose of GSK3380154 (ambrisentan/tadalafil-FDC), GSK1325760 
(ambrisentan) and GF196960 (tadalafil) greater than the protocol defined dose and within 
a 24 hour time period will be considered an overdose.
 
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GSK does not recommend specific treatment for an overdose.
Advice for the investigator is included in the product label for GSK1325760 
(ambrisentan) and GF196960 (tadalafil).
In the event of an overdose the investigator should:
16. Contact the Medical Monitor immediately
17. Closely monitor the subject for adverse events (AEs)/serious adverse events (SAEs) 
and laboratory abnormalities until compound number/name can no longer be 
detected systemically (at least 3 days for compound number/name)
18. Obtain a plasma sample for pharmacokinetic (PK) analysis if requested by the 
Medical Monitor (determined on a case-by-case basis)
19. Document the quantity of the excess dose as well as the duration of the overdosing in 
the CRF.
Decisions regarding dose interruptions or modifications will be made by the investigator 
in consultation with the Medical Monitor based on the clinical evaluation of the subject.
6.9.
Treatment after the End of the Study
Subjects will not receive any additional treatment from GSK after completion of the 
study because only healthy subjects are eligible for study participation.
6.10.
Lifestyle and/or Dietary Restrictions
6.10.1.
Meals and Dietary Restrictions

Refrain from consumption of red wine, Seville oranges, grapefruit or grapefruit 
juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or fruit juices from 
7 days prior to the first dose of study medication until after the final dose.

Dependent on the utility of the dose session subjects will be asked to fast or 
consume the FDA full fat breakfast prior to dosing (EMEA, 2010)

Fasting subjects will be required to fast from midnight before each full in-house 
dosing day (i.e. Day 1) with the exception of water, which will be allowed freely 
except for 1 hour either side of dosing. Subjects will be required to fast up to 4h 
hour post dose on Day 1 of each dose

For treatments given under fed conditions, a standard high-fat breakfast will be 
provided before dosing. See Section 12.4 for details of this meal. This meal 
should be eaten in its entirety within 30 minutes.  The amount consumed will be 
recorded within the source documents and the CRF by the site staff.  Subjects 
will be dosed within 5 minutes of completing the breakfast.  

Subjects should take each dose of investigational product with 240 ml (8 fl oz) of 
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6.10.2.
Caffeine, Alcohol, and Tobacco

During each dosing session, subjects will abstain from ingesting caffeine- or 
xanthine-containing products (e.g., coffee, tea, cola drinks and chocolate) for 24 
hours prior to the start of dosing until collection of the final pharmacokinetic sample 
during each session. 

During each dosing session, subjects will abstain from alcohol for 24 hours prior to 
the start of dosing until collection of the final pharmacokinetic sample during each 
session. 

Smoking is not allowed for 24 hours prior to dosing and whilst subjects are in the 
clinic.
6.10.3.
Activity
Subjects will abstain from strenuous exercise from Screening until Follow-Up.  Subjects 
may participate in light recreational activities during studies (e.g., watch television, read).
6.11.
Concomitant Medications and Non-Drug Therapies
6.11.1.
Permitted Medications and Non-Drug Therapies
Paracetamol at doses of 2 grams/day is permitted for use any time during the study. 
Other concomitant medication may be considered on a case by case basis by the 
Investigator in consultation with the Medical Monitor if required.
6.11.2.
Prohibited Medications and Non-Drug Therapies
Subjects must abstain from taking prescription or non-prescription drugs (including 
vitamins and dietary or herbal supplements), within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose of 
study medication until completion of the Follow-Up visit, unless in the opinion of the 
Investigator and sponsor the medication will not interfere with the study.
7.
STUDY ASSESSMENTS AND PROCEDURES
Protocol waivers or exemptions are not allowed with the exception of immediate safety 
concerns. Therefore, adherence to the study design requirements, including those 
specified in the Time and Events Table, are essential and required for study conduct.
This section lists the procedures and parameters of each planned study assessment.  The 
exact timing of each assessment is listed in the Time and Events Table Section 7.1
The following points must be noted: 

If assessments are scheduled for the same nominal time, THEN the assessments 
should occur in the following order:
1. 12-lead ECG
2. Vital signs
 
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3. Blood draws.  
Note: The timing of the assessments should allow the blood draw to occur at the 
exact nominal time.

The timing and number of planned study assessments, safety and 
pharmacokinetic assessments may be altered during the course of the study 
based on newly available data (e.g., to obtain data closer to the time of peak 
plasma concentrations) to ensure appropriate monitoring. 

The change in timing or addition of time points for any planned study 
assessments must be documented in a Note to File which is approved by the 
relevant GSK study team member and then archived in the study sponsor and 
site study files, but this will not constitute a protocol amendment. 

The IRB/IEC will be informed of any safety issues that require alteration of the 
safety monitoring scheme or amendment of the Informed Consent Form. 
No more than 500 mL of blood will be collected over the duration of the study, including 
any extra assessments that may be required.
 
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7.1.
Time and Events Table
Procedure
Screen
Part 1, 2 and 3A and 3B. Each dose in each Part repeats this schedule.
FU
Notes
Day
≤-31
-1
1
2
3
4
≥7-14
Time (hrs)
Pre-
dose
0
0.5
1
1.5
2
2.5
4
8
12
18
24
36
48
72
Outpatient visit
x
x
x
Admission to unit
x
Informed consent
x
Inclusion and exclusion criteria
x
x
Demography
x
Full physical exam including 
height and weight
x
Brief Physical
x
Medical history (includes 
substance usage)
x
HIV, Hep B and Hep C screen] 
x
Laboratory assessments (include 
liver chemistries) 
x
x
x
x
Only Screening labs need to be taken in 
fasted state.
Serum  hCG Pregnancy test
x
x
Female subjects only
Urine  hCG Pregnancy test
x
Female subjects only
Breathalyser and Smokerlyzer
x
x
DOA testing
x
x
12-lead ECG
x
x
x
x
x
x
x
x
x
Triplicate at screen and baseline, single 
measure at other times, unless out of 
range then triplicates should be
performed
Vital signs
x
x
x
x
x
x
x
x
x
x
x
x
24hr Holter
x
Randomisation
x
Randomised prior to first dose only
Study Treatment
x
AE/SAE review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
SAEs from Screen. AEs from first dose
Concomitant medication review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
PK Sample
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Discharge from Unit
x
For logistical reasons subjects may 
remain in-unit for the 72 hr assessments
if they prefer.
 
 
 
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7.2.
Screening and Critical Baseline Assessments
All subjects must give written consent to participate in this trial.  Consent for screening 
evaluations may be obtained using the ICF for the Hammersmith Medicines Research 
(HMR) healthy subject’s panel, which has been approved by the HRA’s Phase 1 Advert 
Review group.  The study-specific information and consent form will be signed by the 
subject either before any screening evaluation or after the investigator confirms the 
eligibility of the subject for the study and before the subject is randomised to receive the 
first administration of IMP.  Before giving consent, subjects must read the information 
sheet about the study.  They must also read the consent form.  They will then discuss the 
study with the investigator or his deputy and be given the opportunity to ask questions.  
The study-specific information sheet and the consent form must be approved by the REC.
7.2.1.
Demographic/Medical History Assessments
Prior to enrolling in the study and having any study procedures completed, subjects must 
sign the informed consent.
During the Screening visit, each subject will undergo the assessments to determine 
eligibility for enrolment as detailed in Section 5. 
The following demographic parameters will be captured: date of birth, gender, race and 
ethnicity.
Medical/medication/alcohol history will be assessed as related to the eligibility criteria 
listed in Section 5.
7.3.
Pharmacokinetics
7.3.1.
Blood Sample Collection
Blood samples for pharmacokinetic (PK) analysis of ambrisentan and tadalafil will be 
collected at the time points indicated in Section 7.1, Time and Events Table.  The actual 
date and time of each blood sample collection will be recorded.  The timing of PK 
samples may be altered and/or PK samples may be obtained at additional time points to 
ensure thorough PK monitoring.
For analysis of ambrisentan 2.7 mL of blood will be collected into sodium citrate tubes 
and for analysis of tadalafil 2.0 mL of blood will be collected into K2-EDTA tubes. 
Processing, storage and shipping procedures are provided in the Study Reference Manual 
(SRM).
7.3.2.
Sample Analysis
Plasma analysis will be performed under the control of Platform Technology and Science 
In vitro/In vivo Translation (PTS IVIVT) and Third Party Resource, GlaxoSmithKline, 
the details of which will be included in the Study Reference Manual (SRM).
Concentrations of ambrisentan and tadalafil will be determined in plasma samples using 
 
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the currently approved bioanalytical methodology. Raw data will be archived at the 
bioanalytical site (detailed in the SRM).
Once the plasma has been analyzed for ambrisentan and tadalafil any remaining 
plasma may be analyzed for other compound-related metabolites and the results reported 
under a separate PTS-IVIVT, GlaxoSmithKline protocol.
7.4.
Safety
Planned time points for all safety assessments are listed in the Time and Events Table 
(Section 7.1).  Additional time points for safety tests such as vital signs, physical 
examinations and laboratory safety assessments may be added during the course of the 
study based on newly available data to ensure appropriate safety monitoring.
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)
The definitions of an AE or SAE can be found in Appendix 3: Definition of and 
Procedures for Recording, Evaluating, Follow-Up and Reporting of Adverse Events.
The investigator and their designees are responsible for detecting, documenting and 
reporting events that meet the definition of an AE or SAE. 
7.4.1.1.
Time period and Frequency for collecting AE and SAE information

Any SAEs assessed as related to study participation (e.g., protocol-mandated 
procedures, invasive tests, or change in existing therapy) or related to a GSK 
product will be recorded from the time a subject consents to participate in the 
study up to and including any follow up contact. 

AEs will be collected from the start of Study Treatment until the follow up contact 
(see Section 7.4.1.3), at the timepoints specified in the Time and Events Table 
(Section 7.1).

Medical occurrences that begin prior to the start of study treatment but after 
obtaining informed consent may be recorded on the Medical History/Current 
Medical Conditions section of the CRF.

All SAEs will be recorded and reported to GSK within 24 hours, as indicated in 
Section 12.3.

Investigators are not obligated to actively seek AEs or SAEs in former study 
subjects. However, if the investigator learns of any SAE, including a death, at any 
time after a subject has been discharged from the study, and he/she considers the 
event reasonably related to the study treatment or study participation, the 
investigator must promptly notify GSK.
NOTE: The method of recording, evaluating and assessing causality of AEs and SAEs 
plus procedures for completing and transmitting SAE reports to GSK are provided in 
Section 12.3.
 
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7.4.1.2.
Method of Detecting AEs and SAEs
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended 
and non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrence.  Appropriate questions include:

“How are you feeling?” 

“Have you had any (other) medical problems since your last visit/contact?”

“Have you taken any new medicines, other than those provided in this study, 
since your last visit/contact?
7.4.1.3.
Follow-up of AEs and SAEs
After the initial AE/SAE report, the investigator is required to proactively follow each 
subject at subsequent visits/contacts.  All SAEs, and non-serious AEs of special interest 
(as defined in Section 4.6.1) will be followed until resolution, until the condition 
stabilizes, until the event is otherwise explained, or until the subject is lost to follow up 
(as defined in Section 5.4). Further information on follow up procedures is given in Study 
Assessments and Procedures.
7.4.1.4.
Regulatory Reporting Requirements for SAEs
Prompt notification by the investigator to GSK of SAEs related to study treatment (even 
for non- interventional post-marketing studies) is essential so that legal obligations and 
ethical responsibilities towards the safety of subjects and the safety of a product under 
clinical investigation are met. 
GSK has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of a product under clinical investigation.  GSK will 
comply with country specific regulatory requirements relating to safety reporting to the 
regulatory authority, Institutional Review Board (IRB)/Independent Ethics Committee 
(IEC) and investigators.
Investigator safety reports are prepared for suspected unexpected serious adverse 
reactions according to local regulatory requirements and GSK policy and are forwarded 
to investigators as necessary.
An investigator who receives an investigator safety report describing a SAE(s) or other 
specific safety information (e.g., summary or listing of SAEs) from GSK will file it with 
the IB and will notify the IRB/IEC, if appropriate according to local requirements.
7.4.2.
Physical Exams

A complete physical examination will include, at a minimum, assessment of the 
Cardiovascular, Respiratory, Gastrointestinal and Neurological systems. Height 
and weight will also be measured and recorded. 

A brief physical examination will include, at a minimum assessments of the skin, 
lungs, cardiovascular system, and abdomen (liver and spleen).
 
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
Investigators should pay special attention to clinical signs related to previous 
serious illnesses
7.4.3.
Vital Signs

Vital signs will be measured in semi-supine position after 5 minutes rest and will 
include temperature, systolic and diastolic blood pressure and pulse rate and 
respiratory rate. 

Three readings of blood pressure and pulse rate will be taken at Screening and 
baseline (pre-dose). All subsequent assessments will be single measures, unless 
the subjects blood pressure or pulse rate has changed from baseline by >15% and 
then 2 further readings should be taken and recorded:

For triplicate readings:

All 3 readings will be recorded in the CRF

First reading should be rejected

Second and third readings should be averaged to give the measurement to 
be used and this will also be recorded in the CRF
7.4.4.
Electrocardiogram (ECG)

ECGs will be measured in semi-supine position after 5 minutes rest

Triplicate 12-lead ECGs will be obtained at Screening and baseline (predose). All 
subsequent assessments will be a single measures, unless withdrawal criteria are 
meet and in which case 2 further readings should be taken to confirm if 
withdrawal is required. An ECG machine that automatically calculates the heart 
rate and measures PR, QRS, QT, and QTc.  Refer to Section 5.4.2 for QTc 
withdrawal criteria and additional QTc readings that may be necessary.

24 hour continuous cardiac telemetry (Holter) will be performed at Screening.  
Full disclosures will be reviewed in detail and the review maintained as part of the 
subject’s source documents
7.4.5.
Clinical Safety Laboratory Assessments
All protocol required laboratory assessments, as defined in Table 8, must be conducted 
in accordance with the Laboratory Manual, and Protocol Time and Events Schedule.
Laboratory requisition forms must be completed and samples must be clearly labelled 
with the subject number, protocol number, site/centre number, and visit dat.  Reference 
ranges for all safety parameters will be provided to the site by the laboratory responsible 
for the assessments.
If additional non-protocol specified laboratory assessments are performed at the 
institution’s local laboratory and result in a change in subject management or are 
considered clinically significant by the investigator (e.g., SAE or AE or dose 
modification) the results must be recorded in the CRF.
 
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Refer to the SRM for appropriate processing and handling of samples to avoid duplicate 
and/or additional blood draws.
All study-required laboratory assessments will be performed by a local laboratory.
Haematology, clinical chemistry, urinalysis and additional parameters to be tested are 
listed in Table 8. 
Table 8
Protocol Required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Haematology
Platelet Count
RBC Indices:
WBC count with Differential:
RBC Count
MCV
Neutrophils
Hemoglobin
MCH
Lymphocytes
Hematocrit
MCHC
Monocytes
Eosinophils
Basophils
Clinical 
Chemistry 1
BUN
Potassium
AST (SGOT)
Total and direct bilirubin
Creatinine
Sodium
ALT (SGPT)
Total Protein
Glucose 
Calcium
Alkaline phosphatise
Albumin
Routine 
Urinalysis3
As a minimum, but not limited to the following tests and dependent on 
standard urinalysis dipstick used:

Specific gravity

pH, glucose, protein, blood and ketones by dipstick

Microscopic examination (if blood or protein is abnormal)
Other 
Screening 
Tests

HIV

Hepatitis B (HBsAg)

Hepatitis C (Hep C antibody)

Alcohol and drug screen (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines)

Serum  or urine hCG Pregnancy test2
NOTES :
20. Details of Liver Chemistry Stopping Criteria and Required Actions and Follow-Up 
Assessments after liver stopping or monitoring event are given in Section 5.4.1 and 
Appendix 2
21. Local urine testing will be standard for the protocol unless serum testing is required 
by local regulation or ethics committee.
22. Routine Urinalysis results will not be databased, unless a result is out of range and 
clinically significant then it would be captured as an AE  
 
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All laboratory tests with values that are considered clinically significantly abnormal 
during participation in the study or within7 days after the last dose of study treatment 
should be repeated until the values return to normal or baseline.  If such values do not 
return to normal within a period judged reasonable by the investigator, the etiology
should be identified and the sponsor notified.
8.
DATA MANAGEMENT

Data will be double-entered into a clinical database management system 
(ClinPlus Version 3.3).

Management of clinical data will be performed in accordance with applicable 
HMR standards and data cleaning procedures to ensure the integrity of the data, 
e.g., removing errors and inconsistencies in the data.

Adverse events and concomitant medications terms will be coded using 
MedDRA (Medical Dictionary for Regulatory Activities) and an internal 
validated medication dictionary, GSKDrug.  

Original CRFs will be retained by GSK, while HMR will retain a copy. Subject 
initials will not be collected or transmitted to GSK according to GSK policy 
9.
STATISTICAL CONSIDERATIONS AND DATA 
ANALYSES
9.1.
Hypotheses
No formal hypothesis will be tested for study Part 1 and Part 2. An estimation approach 
will be used to (i) estimate the bioavailability of the FDC formulations relative to the 
monotherapies taken concurrently, (ii) for each primary pharmacokinetic endpoint, point 
estimates and corresponding 90% confidence intervals will be constructed for the ratio of 
the geometric mean of the test treatment (FDC formulations) to the geometric mean of 
the reference treatment (co-administration of ambrisentan and tadalafil), (test)/ 
(reference).  The objective of Part 2 is to assess whether differences in granulation size 
impact the pharmacokinetics of ambrisentan and tadalafil; the estimation approach for 
bioavailability is therefore applicable to Part 2.
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 10 
mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 10 mg & 
tadalafil 40 mg taken concurrently (reference) in healthy subjects in both the fed and 
fasted states. The null hypothesis is that the true ratio of the geometric mean of the test 
treatment to the geometric mean of the reference treatment, (test)/(reference), for each 
primary PK endpoint, is either less than 0.80 or greater than 1.25.  The alternate 
hypothesis is that the true ratio of the test treatment geometric mean to the reference 
treatment geometric mean is at least 0.80 and no greater than 1.25.  Symbolically, this is 
expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
 
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i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e., treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test (TOST) 
procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be used to test 
this set of hypotheses. This is equivalent to requiring that a 90% interval for the true ratio 
of test to reference geometric means falls entirely within the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test (1) the bioequivalence of 
a FDC of ambrisentan 5 mg + tadalafil 40 mg (test) relative to reference monotherapies 
of ambrisentan 5 mg & tadalafil 40 mg taken concurrently (reference) and (2) the 
bioequivalence of a FDC of ambrisentan 5 mg + tadalafil 20 mg (test) relative to 
reference monotherapies of ambrisentan 5 mg & tadalafil 20 mg taken concurrently 
(reference), in healthy subjects in the fasted states. Therefore, the analysis approach for 
bioequivalence for Part 3A is applicable to Part 3B.
9.2.
Sample Size Considerations
9.2.1.
Sample Size Assumptions
For Part 1 and Part 2 of the study, the sample size assumptions are based on previously 
reported estimates of within subject CV for AUC(0-∞) and Cmax for ambrisentan (GS-US-
300-0112, 2008) and tadalafil (Forgue, 2005). Table 9 summarizes the estimates of 
within subject CV for the primary endpoints AUC (0-∞) and Cmax.
Table 9
Estimates of within subject CV for the primary end points AUC (0-∞) 
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
For Part 3A of the study, Table 10 summarized the observed estimates of between and 
within subject CV and observed ratio of geometric means for the primary endpoints 
AUC(0-t) and Cmax from Part 1 of the study. 
 
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Table 10
Estimates of within subject CV for the primary end points AUC (0-∞)
and Cmax for ambrisentan and tadalafil from part 1 of the study
CVw: within subject CV
ambrisentan 
tadalafil 
Cmax
22.42%
14.53%
AUC (0-∞)
8.12%
10.32%
The largest of the within subject CV estimates 22.42% translates to a standard deviation 
(SD) of 0.221 on the natural log scale.  Based on this SD, a sample size of 23 statistically 
evaluable subjects will have 90% power to demonstrate bioequivalence. This calculation 
assumes:

a true ratio of 1,

the within-subject variability from the current study will not be larger than that 
used in the sample size calculations,

data are log-normally distributed, and each t-test is made at the 5% level.
For Part 3B of the study, same assumptions are adopted as Part 3A. Thus, a sample size 
of 23 statistically evaluable subjects will have 90% power to demonstrate bioequivalence.  
However, if the within subject CVs obtained after Part 2 are significantly higher than the 
ones in Part 1 (shown in Table 10), a sample size re-esimation may be conducted to 
adjust the sample size for Part 3A and 3B to ensure the study maintain enough power to 
demonstrate bioequivalence. 
9.2.2.
Sample Size Sensitivity
For Part 1 and Part 2, assuming the within subject CV differs from 22% and the number 
of statistically evaluable subjects is lower than 20, then Table 11 shows the precision (the 
half width of the 90% confidence interval) for a range of evaluable subjects.
 
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Table 11
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
For Part 3, based on the observed data from previous studies, the ratio of geometric mean 
for ambrisentan Cmax is unlikely to be 1. The effects on the power of declaring 
bioequivalence in the face of a shift in the expected ratio of the geometric means were 
examined.
If, under all other assumptions outlined above, the actual ratio of geometric means for 
ambrisentan Cmax is 0.97, then 26 evaluable subjects are needed to have 90% power to 
conclude bioequivalence.
With 26 evaluable subjects, the current study design has at least 90% power to conclude 
bioequivalance (with both Cmax and AUC for ambrisentan and tadalafil).
9.2.3.
Sample Size Re-estimation or Adjustment
No sample size re-estimation will be performed.  However, if the estimation of the within 
subject CV from Part 2 of the study is much larger than the within subject CV from Part 1 
of the study, the sample size for Part3 may be re-evaluated. 
9.3.
Data Analysis Considerations
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
sampling times recorded during the study although supplementary analayis will be 
available based on the nominal times.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-life 
(t1/2).  
 
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Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in fixed effect ANOVA model and mixed effects 
model.
9.3.1.
Analysis Populations
9.3.1.1.
Safety Population
All subjects enrolled into the study who have received at least one dose of investigational
product will be included in the Safety Population.
9.3.1.2.
Pharmacokinetic Concentration Population
The PK Concentration Population will include all subjects for whom at least one PK 
sample was obtained and analyzed.
9.3.1.3.
Pharmacokinetic Parameter Population
For each PK parameter, the PK Parameter Population will include all subjects who 
provide PK parameter data.
9.3.2.
Interim Analysis
No formal interim analysis is planned. However, ongoing analyses of pharmacokinetic 
data will be performed in Part 1 and Part 2 of the study in order to direct development of 
the FDC for later parts of the study.  Headline results based on statistical analysis using 
preliminary pharmacokinetic data with nominal time may be produced when 80% and 
100% subjects complete Part 1 and Part 2 of the study to assist development of FDCs.  
Treatment and period information from the crossover design may be used in the analysis; 
sequence information may also be included depending on availability. 
9.4.
Key Elements of Analysis Plan
9.4.1.
Pharmacokinetic Analyses
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D. 
 
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Data will be listed and summarized according to GlaxoSmithKline reporting standards, 
where applicable.  Listings will be sorted by subject, period, noting treatment.  
Summaries will be presented by treatment.  Unless stated otherwise, descriptive 
summaries will include n, mean, standard deviation (SD), coefficient of variation (CV), 
median, minimum, and maximum for continuous variables, n and percent for categorical 
variables and geometric mean, 95% confidence interval (CI), and the between-subject CV 
(CVb) based on geometric mean for the log-transformed PK parameters.
Version 9.3 (or higher) of the SAS system will be used for statistical analysis of the data
as well as to generate tables, figures, and listings.
Any deviation(s) from the original analyses planned in the protocol will be reported in the 
Reporting and Analysis Plan (RAP) and/or in the Clinical Pharmacology Study Report.
The PK parameters, AUC(0-), AUC(0-t), Cmax, and t½ will be transformed using natural 
logarithms. Missing PK parameters will not be imputed.
All data from withdrawn subjects will be listed.
No adjustment for multiple tests or comparisons is planned.
Actual visit day will be used for safety assessments. For the assessment of 
bioequivalence, both actual time of sampling, and nominal (planned) sampling time, will 
be used to estimate PK parameters at the end of the study. PK parameters estimation 
based on nominal sampling time will be used for generating headline results. According 
to EMEA guidilines on GUIDELINE ON THE INVESTIGATION OF 
BIOEQUIVALENCE (2010, Doc. Ref.:[CPMP/EWP/QWP/1401/98 Rev. 1/ Corr **. 
2010]), following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using fixed effect ANOVA model with fixed 
effect terms for sequence, subject within sequence, period and treatment (formulation).
As a sensitivity analysis, mixed effect model with fixed effect terms for period, sequence 
and treatment (formulation), and subject as random effect term (i.e. ANOVA method 
using period, sequence and treatment as fixed effect, and subject as random effect) will 
be performed.
Point estimates and their associated 90% confidence intervals will be calculated for the 
differences: F1 - R, F2 - R, F3 -R, F4 - R for Part 1; FG1-R, FG2-R, FG3-R for Part 2; 
X1-R1, X2-R2 for Part 3A; and Y1-R3, Y2-R4 for Part 3B. The point estimates and their 
associated 90% confidence intervals will then be back-transformed to provide point 
estimates and 90% confidence intervals for the ratios, F1:R, F2:R, F3:R, F4:R; for Part 1; 
FG1:R, FG2:R, FG3:R for Part 2; X1:R1,X2:R2 for Part3A; and Y1:R3 and Y2:R4 for 
Part 3B.
9.4.2.
Safety Analyses
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
 
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10.
STUDY GOVERNANCE CONSIDERATIONS
10.1.
Posting of Information on Publicly Available Clinical Trial 
Registers
Study information from this protocol will be posted on publicly available clinical trial 
registers before enrollment of subjects begins.
10.2.
Regulatory and Ethical Considerations, Including the 
Informed Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.
The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent

Investigator reporting requirements (e.g. reporting of AEs/SAEs/protocol 
deviations to IRB/IEC)

GSK will provide full details of the above procedures, either verbally, in writing, 
or both.

Signed informed consent must be obtained for each subject prior to participation 
in the study

The IEC/IRB, and where applicable the regulatory authority, approve the clinical 
protocol and all optional assessments, including genetic research.  

Optional assessments (including those in a separate protocol and/or under separate 
informed consent) and the clinical protocol should be concurrently submitted for 
approval unless regulation requires separate submission.  

Approval of the optional assessments may occur after approval is granted for the 
clinical protocol where required by regulatory authorities. In this situation, 
written approval of the clinical protocol should state that approval of optional 
assessments is being deferred and the study, with the exception of the optional 
assessments, can be initiated.
 
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10.3.
Quality Control (Study Monitoring)

In accordance with applicable regulations including GCP, and GSK procedures, 
GSK monitors will contact the site prior to the start of the study to review with the 
site staff the protocol, study requirements, and their responsibilities to satisfy 
regulatory, ethical, and GSK requirements.  

When reviewing data collection procedures, the discussion will also include 
identification, agreement and documentation of data items for which the CRF will 
serve as the source document.
GSK will monitor the study and site activity to verify that the:

Data are authentic, accurate, and complete.

Safety and rights of subjects are being protected.

Study is conducted in accordance with the currently approved protocol and any 
other study agreements, GCP, and all applicable regulatory requirements.
The investigator and the head of the medical institution (where applicable) agrees to 
allow the monitor direct access to all relevant documents
10.4.
Quality Assurance

To ensure compliance with GCP and all applicable regulatory requirements, GSK 
may conduct a quality assurance assessment and/or audit of the site records, and 
the regulatory agencies may conduct a regulatory inspection at any time during or 
after completion of the study. 

In the event of an assessment, audit or inspection, the investigator (and institution) 
must agree to grant the advisor(s), auditor(s) and inspector(s) direct access to all 
relevant documents and to allocate their time and the time of their staff to discuss 
the conduct of the study, any findings/relevant issues and to implement any 
corrective and/or preventative actions to address any findings/issues identified.
10.5.
Study and Site Closure

Upon completion or premature discontinuation of the study, the GSK monitor will 
conduct site closure activities with the investigator or site staff, as appropriate, in 
accordance with applicable regulations including GCP, and GSK Standard 
Operating Procedures.

GSK reserves the right to temporarily suspend or prematurely discontinue this 
study at any time for reasons including, but not limited to, safety or ethical issues 
or severe non-compliance. For multicenter studies, this can occur at one or more 
or at all sites.  

If GSK determines such action is needed, GSK will discuss the reasons for taking 
such action with the investigator or the head of the medical institution (where 
applicable).  When feasible, GSK will provide advance notification to the 
investigator or the head of the medical institution, where applicable, of the 
impending action.
 
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
If the study is suspended or prematurely discontinued for safety reasons, GSK will 
promptly inform all investigators, heads of the medical institutions (where 
applicable) and/or institution(s) conducting the study. GSK will also promptly 
inform the relevant regulatory authorities of the suspension or premature 
discontinuation of the study and the reason(s) for the action.  

If required by applicable regulations, the investigator or the head of the medical 
institution (where applicable) must inform the IRB/IEC promptly and provide the 
reason for the suspension or premature discontinuation.
10.6.
Records Retention

Following closure of the study, the investigator or the head of the medical 
institution (where applicable) must maintain all site study records (except for 
those required by local regulations to be maintained elsewhere), in a safe and 
secure location.  

The records must be maintained to allow easy and timely retrieval, when needed 
(e.g., for a GSK audit or regulatory inspection) and must be available for review 
in conjunction with assessment of the facility, supporting systems, and relevant 
site staff.  

Where permitted by local laws/regulations or institutional policy, some or all of 
these records can be maintained in a format other than hard copy (e.g., microfiche, 
scanned, electronic); however, caution needs to be exercised before such action is 
taken.  

The investigator must ensure that all reproductions are legible and are a true and 
accurate copy of the original and meet accessibility and retrieval standards, 
including re-generating a hard copy, if required.  Furthermore, the investigator 
must ensure there is an acceptable back-up of these reproductions and that an 
acceptable quality control process exists for making these reproductions.

GSK will inform the investigator of the time period for retaining these records to 
comply with all applicable regulatory requirements.  The minimum retention time 
will meet the strictest standard applicable to that site for the study, as dictated by 
any institutional requirements or local laws or regulations, GSK 
standards/procedures, and/or institutional requirements.  

The investigator must notify GSK of any changes in the archival arrangements, 
including, but not limited to, archival at an off-site facility or transfer of 
ownership of the records in the event the investigator is no longer associated with 
the site.
10.7.
Provision of Study Results to Investigators, Posting of 
Information on Publically Available Clinical Trials Registers 
and Publication
Where required by applicable regulatory requirements, an investigator signatory will be 
identified for the approval of the clinical study report.  The investigator will be provided 
reasonable access to statistical tables, figures, and relevant reports and will have the 
 
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opportunity to review the complete study results at a GSK site or other mutually-
agreeable location.
GSK will also provide the investigator with the full summary of the study results.  The 
investigator is encouraged to share the summary results with the study subjects, as 
appropriate.
GSK will provide the investigator with the randomization codes for their site only after 
completion of the full statistical analysis.
The procedures and timing for public disclosure of the results summary and for 
development of a manuscript for publication will be in accordance with GSK Policy.
A manuscript will be progressed for publication in the scientific literature if the results 
provide important scientific or medical knowledge.
 
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11.
REFERENCES
Adcirca: EMA Summary of product Characteristics, Eli Lily, last updated 29-Sep-2015 at 
time of publication
EMEA guidelines on clinical development of fixed dose combination medicinal products. 
CHMP/EWP/240/95 Rev.1. 2008
EMEA guidelines on the investigation of bioequivalence. CPMP/EWP/QWP/1401/98 
Rev. 1/ Corr **. 2010
Forgue S, Patterson B, Bedding A, et al. Tadalafil pharmacokinetics in healthy subjects. 
British Journal of Clinical Pharmacology. 20-Dec-2005
Galiè N et al. Initial Use of Ambrisentan plus Tadalafil in Pulmonary Arterial 
Hypertension. N Engl J Med 2015a;373:834-44.
Galiè N, Humbert M, Vachiery J.L et al. 2015 ESC/ERS Guidelines for the diagnosis and 
treatment of pulmonary hypertension. European Heart Journal Advance Access, 
published August 29, 2015b
Ghofrani AH, Galiè N, Grimminger F, et al. N Engl J Med 2013;369:330-40. Riociguat 
for the Treatment of Pulmonary Arterial Hypertension
Gilead Clinical Study report GS-US-300-0112: A Phase 1, Open-Label, Randomized, 
Two-Period, Crossover Study to Assess the Pharmacokinetics of Ambrisentan Alone and 
in the Presence of Multiple Doses of Tadalafil, and to assess the Pharmacokinetics of 
Tadalafil Alone and in the Presence of Multiple Doses of Ambrisentan in Healthy Adult 
Volunteers. Version 1.0, dated 27-June-2008
Gilead Investigator Brochure, Letaris, Volibris, Fifteenth edition: 06 April 2015
GSK Clinical Study Report for study AMB112565, GSK document 2014N193963_00: 
AMBITION: A Randomized, Multicenter Study of First-Line Ambrisentan and Tadalafil 
Combination Therapy in Subjects with Pulmonary Arterial Hypertension., dated 13-Nov-
14
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(a): 24 Table 3-2
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(b): 28.
Liang F, Yang S, Yao L, Belardinelli L, Shryock J. Hypertension. 2012 Mar;59(3):705-
11. Ambrisentan and tadalafil synergistically relax endothelin-induced contraction of rat 
pulmonary arteries.
Pulido T, Adzerikho I, Channick RN, et al. N Engl J Med. 2013 Aug 29;369(9):809-18. 
Macitentan and Morbidity and Mortality in Pulmonary Arterial Hypertension
 
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Schuirmann DJ. A comparison of the two one-sided tests procedure and the power 
approach for assessing the equivalence of average bioavailability, J Pharmacokin. 
Biopharm. 1987:15:657–680. 
Spence R, Mandagere A, Harrison B, et al.  J Pharm Sci. 2009 Dec;98(12):4962-74. No 
clinically relevant pharmacokinetic and safety interactions of ambrisentan in combination 
with tadalafil in healthy volunteers. 
Volibris: EMA Summary of Product Characteristics, GSK, variation II/41, at time of 
publication, 2015.
 
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12.
APPENDICES
12.1.
Appendix 1: Abbreviations and Trademarks
Abbreviations
AE
Adverse Event
API
Active Pharmaceutical Ingredient
AUC
Area Under Curve
BE
Bioequivalence
CDISC
Clinical Data Interchange Standards Consortium
cGMP
Cyclic guanosine monophosphate
CI
Confidence Interval
Cmax
Maximum concentration
CV
Coefficient of variance 
CVw
Coefficient of variance within subject
EMA
European Medicines Agency 
ERA
Endothelin Receptor Antagonists
ERS
European Respiratory Society 
ESC
European Society of Cardiology
ET-1
Endothelin Receptor - 1
EU
European Union
FDA
Federal Drug Agency
FDC
Fixed Dose Combination
NAION
Non-arteritic anterior ischemic optic neuropathy
NO
Nitrous Oxide
PDE-5
Phosphodiesterase type 5
SAE
Serious Adverse Events
SRM
Study Reference Manual
t1/2
Half Life
tmax
Time to maximum concentration
WHO
World Health Organisation
Trademark Information
Trademarks of the GlaxoSmithKline 
group of companies
Trademarks not owned by the 
GlaxoSmithKline group of companies
NONE
Adcirca
WinNonlin
 
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12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments
Phase I Liver chemistry stopping criteria have been designed to assure subject safety and 
to evaluate liver event aetiology (in alignment with the FDA premarketing clinical liver 
safety guidance).
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf.
Phase I liver chemistry stopping criteria and required follow up assessments
Liver Chemistry Stopping Criteria – Liver Stopping Event
ALT-absolute
ALT≥3xULN
If  ALT≥3xULN  AND bilirubin1,2 2xULN (>35% direct bilirubin) or  INR >1.5, 
Report as an SAE.
See additional Actions and Follow-Up Assessments listed below
Required Actions and Follow up Assessments following Liver Stopping Event
Actions
Follow-Up Assessments

Immediately discontinue study treatment 

Report the event to GSK within 24 hours

Complete the liver event CRF, and complete 
an SAE data collection tool if the event also 
meets the criteria for an SAE2

Perform liver event follow up assessments 

Monitor the subject until liver chemistries 
resolve, stabilise, or return to within baseline 
(see MONITORING below)
MONITORING:
If ALT≥3xULN AND bilirubin 2xULN  or INR 
>1.5

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within  24 
hrs

Monitor subjects twice weekly until liver 
chemistries resolve, stabilise or return to 
within baseline

Viral hepatitis serology3

Blood sample for pharmacokinetic (PK) 
analysis, obtained within 96hrs of last dose

Serum creatine phosphokinase (CPK) and 
lactate dehydrogenase (LDH).

Fractionate bilirubin, if total 
bilirubin2xULN

Obtain complete blood count with 
differential to assess eosinophilia

Record the appearance or worsening of 
clinical symptoms of liver injury, or 
hypersensitivity, on the AE report form

Record use of concomitant medications on 
the concomitant medications report form 
including acetaminophen, herbal remedies, 
other over the counter medications.

Record alcohol use on the liver event 
alcohol intake case report form
If ALT≥3xULN AND bilirubin 2xULN  or 
 
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Liver Chemistry Stopping Criteria – Liver Stopping Event

A specialist or hepatology consultation is 
recommended
If ALT≥3xULN AND bilirubin < 2xULN  and 
INR ≤1.5:

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within 24-
72 hrs

Monitor subjects weekly until liver 
chemistries resolve, stabilize or return to 
within baseline
INR >1.5:

Anti-nuclear antibody, anti-smooth muscle 
antibody, Type 1 anti-liver kidney 
microsomal antibodies, and quantitative 
total immunoglobulin G (IgG or gamma 
globulins).

Serum acetaminophen adduct HPLC assay 
(quantifies potential acetaminophen 
contribution to liver injury in subjects with 
definite or likely acetaminophen use in the 
preceding week [James, 2009].

Liver imaging (ultrasound, magnetic 
resonance, or computerised tomography) 
and /or liver biopsy to evaluate liver 
disease; complete Liver Imaging and/or 
Liver Biopsy CRF forms.
1.
Serum bilirubin fractionation should be performed if testing is available. If serum bilirubin fractionation is not 
immediately available, discontinue study treatment for that subject if ALT 3xULN and bilirubin 2xULN. 
Additionally, if serum bilirubin fractionation testing is unavailable, record presence of detectable urinary bilirubin on 
dipstick, indicating direct bilirubin elevations and suggesting liver injury. 
2.
All events of ALT 3xULN and bilirubin 2xULN (>35% direct bilirubin) or ALT 3xULN and INR>1.5, if INR 
measured, which may indicate severe liver injury (possible ‘Hy’s Law’), must be reported as an SAE (excluding 
studies of hepatic impairment or cirrhosis); INR measurement is not required and the threshold value stated will 
not apply to subjects receiving anticoagulants
3.
Includes: Hepatitis A IgM antibody; Hepatitis B surface antigen and Hepatitis B Core Antibody (IgM); Hepatitis C
RNA; Cytomegalovirus IgM antibody;  Epstein-Barr viral capsid antigen IgM antibody (or if unavailable, obtain 
heterophile antibody or monospot testing);  Hepatitis E IgM antibody 
4.
PK sample may not be required for subjects known to be receiving placebo or non-GSK comparator treatments.  
Record the date/time of the PK blood sample draw and the date/time of the last dose of study treatment prior to 
blood sample draw on the CRF. If the date or time of the last dose is unclear, provide the subject’s best 
approximation. If the date/time of the last dose cannot be approximated OR a PK sample cannot be collected in 
the time period indicated above, do not obtain a PK sample. Instructions for sample handling and shipping are in 
the SRM. 
References
James LP, Letzig L, Simpson PM, Capparelli E, Roberts DW, Hinson JA, Davern TJ, Lee 
WM. Pharmacokinetics of Acetaminophen-Adduct in Adults with Acetaminophen 
Overdose and Acute Liver Failure. Drug Metab Dispos 2009; 37:1779-1784.
 
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12.3.
Appendix 3: Definition of and Procedures for Recording, 
Evaluating, Follow-Up and Reporting of Adverse Events
12.3.1.
Definition of Adverse Events
Adverse Event Definition:

An AE is any untoward medical occurrence in a patient or clinical investigation 
subject, temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product.

NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal product.
Events meeting AE definition include:

Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (e.g., ECGs, radiological scans, vital signs 
measurements), including those that worsen from baseline, and felt to be clinically 
significant in the medical and scientific judgement of the investigator.

Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

New conditions detected or diagnosed after study treatment administration even 
though it may have been present prior to the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected interaction.

Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication (overdose per se will not be reported as an 
AE/SAE unless this is an intentional overdose taken with possible suicidal/self-
harming intent.  This should be reported regardless of sequelae).
Events NOT meeting definition of an AE include:

Any clinically significant abnormal laboratory findings or other abnormal safety 
assessments which are associated with the underlying disease, unless judged by the 
investigator to be more severe than expected for the subject’s condition.

The disease/disorder being studied or expected progression, signs, or symptoms of 
the disease/disorder being studied, unless more severe than expected for the subject’s 
condition.

Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that 
leads to the procedure is an AE.

Situations where an untoward medical occurrence did not occur (social and/or 
 
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convenience admission to a hospital).

Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen.
12.3.2.
Definition of Serious Adverse Events
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, 
death due to progression of disease, etc).
Serious Adverse Event (SAE) is defined as any untoward medical occurrence that, 
at any dose:
a.
Results in death
b.
Is life-threatening
NOTE:
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
subject was at risk of death at the time of the event.  It does not refer to an event, which 
hypothetically might have caused death, if it were more severe.
c.
Requires hospitalization or prolongation of existing hospitalization
NOTE:

In general, hospitalization signifies that the subject has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for 
observation and/or treatment that would not have been appropriate in the physician’s 
office or out-patient setting.  Complications that occur during hospitalization are 
AEs.  If a complication prolongs hospitalization or fulfills any other serious criteria, 
the event is serious.  When in doubt as to whether “hospitalization” occurred or was 
necessary, the AE should be considered serious.

Hospitalization for elective treatment of a pre-existing condition that did not worsen 
from baseline is not considered an AE.
d.
Results in disability/incapacity
NOTE:

The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.

This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, 
and accidental trauma (e.g. sprained ankle) which may interfere or prevent everyday 
life functions but do not constitute a substantial disruption
 
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e.
Is a congenital anomaly/birth defect
f.
Other situations:

Medical or scientific judgment should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be 
immediately life-threatening or result in death or hospitalization but may jeopardize 
the subject or may require medical or surgical intervention to prevent one of the 
other outcomes listed in the above definition.  These should also be considered 
serious.

Examples of such events are invasive or malignant cancers, intensive treatment in an 
emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse
g.
Is associated with liver injury and impaired liver function defined as:

ALT 3xULN and total bilirubin* 2xULN (>35% direct), or

ALT 3xULN and INR** > 1.5.
* Serum bilirubin fractionation should be performed if testing is available; if unavailable, 
measure urinary bilirubin via dipstick.  If fractionation is unavailable and ALT 3xULN 
and total bilirubin 2xULN, then the event is still to be reported as an SAE.
** INR testing not required per protocol and the threshold value does not apply to 
subjects receiving anticoagulants.  If INR measurement is obtained, the value is to be 
recorded on the SAE form.

Refer to Section 12.2 for the required liver chemistry follow up instructions
12.3.3.
Definition of Cardiovascular Events
Cardiovascular Events (CV) Definition:
Investigators will be required to fill out the specific CV event page of the CRF for the 
following AEs and SAEs:

Myocardial infarction/unstable angina

Congestive heart failure

Arrhythmias

Valvulopathy

Pulmonary hypertension

Cerebrovascular events/stroke and transient ischemic attack

Peripheral arterial thromboembolism

Deep venous thrombosis/pulmonary embolism

Revascularization
 
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12.3.4.
Recording of AEs and SAEs
AEs and SAE Recording:

When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) 
relative to the event.

The investigator will then record all relevant information regarding an AE/SAE in 
the CRF

It is not acceptable for the investigator to send photocopies of the subject’s medical 
records to GSK in lieu of completion of the GSK, AE/SAE CRF page.

There may be instances when copies of medical records for certain cases are 
requested by GSK.  In this instance, all subject identifiers, with the exception of the 
subject number, will be blinded on the copies of the medical records prior to 
submission of to GSK.

The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information.  In such cases, the diagnosis will be 
documented as the AE/SAE and not the individual signs/symptoms.

Subject-completed Value Evidence and Outcomes questionnaires and the collection 
of AE data are independent components of the study.

Responses to each question in the Value Evidence and Outcomes questionnaire will 
be treated in accordance with standard scoring and statistical procedures detailed by 
the scale’s developer.

The use of a single question from a multidimensional health survey to designate a 
cause-effect relationship to an AE is inappropriate.
12.3.5.
Evaluating AEs and SAEs
Assessment of Intensity
The investigator will make an assessment of intensity for each AE and SAE reported 
during the study and will assign it to one of the following categories:

Mild:  An event that is easily tolerated by the subject, causing minimal discomfort 
and not interfering with everyday activities.

Moderate:  An event that is sufficiently discomforting to interfere with normal 
everyday activities

Severe:  An event that prevents normal everyday activities. - an AE that is assessed 
as severe will not be confused with an SAE. Severity is a category utilized for rating 
the intensity of an event; and both AEs and SAEs can be assessed as severe.

An event is defined as ‘serious’ when it meets at least one of the pre-defined 
outcomes as described in the definition of an SAE.
 
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Assessment of Causality

The investigator is obligated to assess the relationship between study treatment and 
the occurrence of each AE/SAE.

A "reasonable possibility" is meant to convey that there are facts/evidence or 
arguments to suggest a causal relationship, rather than a relationship cannot be ruled 
out.

The investigator will use clinical judgment to determine the relationship.

Alternative causes, such as natural history of the underlying diseases, concomitant 
therapy, other risk factors, and the temporal relationship of the event to the study 
treatment will be considered and investigated.

The investigator will also consult the Investigator Brochure (IB) and/or Product 
Information, for marketed products, in the determination of his/her assessment.

For each AE/SAE the investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality.

There may be situations when an SAE has occurred and the investigator has minimal 
information to include in the initial report to GSK.  However, it is very important 
that the investigator always make an assessment of causality for every event 
prior to the initial transmission of the SAE data to GSK.

The investigator may change his/her opinion of causality in light of follow up 
information, amending the SAE data collection tool accordingly.

The causality assessment is one of the criteria used when determining regulatory 
reporting requirements.
Follow up of AEs and SAEs

The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as may be indicated or as requested by GSK to 
elucidate as fully as possible the nature and/or causality of the AE or SAE.

The investigator is obligated to assist.  This may include additional laboratory tests 
or investigations, histopathological examinations or consultation with other health 
care professionals.

If a subject dies during participation in the study or during a recognized follow up 
period, the investigator will provide GSK with a copy of any post-mortem findings, 
including histopathology.

New or updated information will be recorded in the originally completed CRF.

The investigator will submit any updated SAE data to GSK within the designated 
reporting time frames.
 
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12.3.6.
Reporting of SAEs to GSK
SAE reporting to GSK via paper CRF

Facsimile transmission of the SAE paper CRF is the preferred method to transmit 
this information to the Medical Monitor or the SAE coordinator

In rare circumstances and in the absence of facsimile equipment, notification by 
telephone is acceptable, with a copy of the SAE data collection tool sent by 
overnight mail

Initial notification via the telephone does not replace the need for the investigator to 
complete and sign the SAE CRF pages within the designated reporting time frames.

Contacts for SAE receipt can be found at this beginning of the protocol on the 
Sponsor/Medical Monitor Contact Information page.
 
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12.4.
Appendix 4 High Fat Meal Content

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
The standard high-fat meal will be the meal suggested by the EMEA guidelines on the 
investigation of bioequivalence 2010. The meal should be a high-fat (approximately 50 
percent of total caloric content of the meal) and high-calorie (approximately 800 to 1000 
kcal) meal. This test meal should derive approximately 150, 250, and 500-600 kcal from 
protein, carbohydrate, and fat, respectively.(EMEA, 2010)
 
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12.5.
Appendix 5: Protocol Changes
12.5.1.
Protocol changes for Amendment 01 (14-Apr-2016) from Original 
Protocol (16-Dec-2015)
The purpose of this amendment is to clarify the hypotension withdrawal criteria in 
section 5.4.3, address changes to GSK medical monitor and minor administrative 
changes.
Summary of Changes
Section 1 PROTOCOL SYNOPSIS FOR STUDY 201964
AND
Section 3 OBJECTIVES AND ENDPOINTS
CHANGE FROM
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assed by vital signs, ECG, clinical laboratory 
safety data and review of adverse events
CHANGE TO
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
 
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Section 5.4.3 Hypotension
CHANGE FROM
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 or diastolic <50 confirmed by triplicate reading 
taken up to 5 minutes apart.
CHANGE TO
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 mm HG and diastolic <50 mm HG confirmed 
by triplicate reading taken up to 5 minutes apart and is judged clinically 
significant and symptomatic by the investigator. 
Section 6.3 Treatment Assignment
ADDITION 
Table 6 Treatment Sequence assignments for Part 1, Part 2 and Part 3
Part 2 and 
Part 3 
merged 
together
5 (Two FDCs selected from 
Part 1)
X1 X2 R Y1 Y2
X2 Y1 X1 Y2 R
Y1 Y2 X2 R X1
Y2 R Y1 X1 X2
R X1 Y2 X2 Y1
Y2 Y1 R X2 X1
R Y2 X1 Y1 X2
X1 R X2 Y2 Y1
X2 X1 Y1 R Y2
Y1 X2 Y2 X1 R
1:1:1:1:1:1:1:1:1:1
3 (One FDC selected from 
Part 1)
X1 X2 R
R X2 X1
X2 R X1
F1 R X2
R X1 X2
X2 X1 R
3:3:3:3:3:3 + 2 random
 
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12.5.2.
Protocol changes for Amendment 2 (08-FEB-2017) from 
Amendment 1(14-Apr-2016)
Amendment 2 Summary of Changes
The purpose of this substantial amendment is to include bioequivalence assessment in 
Part 3 to replace the optional food effect and the addition of new dosage strengths. 
Furthermore GSK medical monitor contact have changed 
List of Specific Changes
Medical Monitor/SAE Contact Information
Rationale for Change: GSK medical monitor contact have changed
CHANGE FROM
Role
Name
Day Time Phone Number and 
email address
After-hours Phone/Cell/
Pager Number
Fax 
Number
Primary 
Medical 
Monitor
Site to 
scan and 
send to 
email
Secondary 
Monitor
Site to 
scan and 
send to 
email
SAE 
contact  
information 
Site to 
scan and 
send to 
email
CHANGE TO
Role
Name
Day Time Phone Number and 
email address
After-hours 
Phone/Cell/
Pager Number
Fax 
Number
Primary 
Medical 
Monitor
Site to 
scan and 
send to 
email
Secondary 
Medical 
Monitor
Site to 
scan 
and 
send to 
email
 
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Role
Name
Day Time Phone Number and 
email address
After-hours 
Phone/Cell/
Pager Number
Fax 
Number
SAE 
contact  
information 
Site to 
scan 
and 
send to 
email
Section 1 - Protocol Synopsis Overall design
CHANGE FROM:
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects.
All subjects will attend the unit for Screening within 31 days of their first dose. Eligible 
subjects will be randomised to order of treatment in that study parts in which they are 
included and the order in which these treatments are administered will be in accordance 
with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose.  The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
 
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However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast
CHANGE TO: 
This is a single centre, Phase 1, single dose, randomised, open label crossover study in 
healthy volunteers with 3 study parts. Part 1 will include a 5 way cross-over. Part 2 
and Part 3 (A&B) will each include a 4 way cross-over. All subjects will attend the 
unit for Screening within 31 days of their first dose. Eligible subjects will be 
randomised to order of treatment in that study parts in which they are included and 
the order in which these treatments are administered will be in accordance with the 
randomisation schedule.
There will be a minimum washout of 7 days between each dose in Parts 1 and 2. In Part 
3 the washout will be at least 10 days.  A Follow-Up Visit should be completed within 
7-14 days of completion of a subject’s last dose.  The utility of each of the 3 study parts is 
as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
 
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Part 2
This study part will include an evaluation of  the bioavailability, safety and 
tolerability of 3 granulation forms of a single FDC (ambrisentan 10mg + tadalafil 40 
mg) identified from Part 1. These data will be compared to that for the reference of 
the 2 monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 
40 mg). All doses are taken in the fasted state.
Part 3 A
Part 3A of the study is set to establish bioequivalence between the candidate FDC 
from Part 2. This study part will have 4 dose sessions and will assess the 
bioequivalence, in both the fed and fasted state, of the FDC against the reference 
ambrisentan 10 mg + tadalafil 40mg monotherapies in the fed and fasted state.  The 
fed arms of this part will have a standard high fat breakfast. (EMEA, 2010).
Part 3 B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 
40mg and ambrisentan 5 mg + tadalafil 20mg). This study part will have 4 dose 
sessions and will assess the bioequivalence of the two FDC dose strengths against the 
reference ambrisentan 5 mg + tadalafil 40mg and ambrisentan 5 mg + tadalafil 
20mg monotherapies in the fasted state.
Section 1 - Protocol Synopsis Treatment Arms and Duration
CHANGE FROM: 
The proposed treatment arms for each study part are described here; however the 
treatments in Parts 2 and 3 may be changed dependent on the utility and results from the 
previous part.
 
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Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil FDC5 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil FDC6 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil FDC7 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC4 (10mg/40mg)
ambrisentan and tadalafil FDC8 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less FDC treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. The minimum will be approximately 3 weeks and the maximum will be 
approximately 18 weeks.
CHANGE TO:
The proposed treatment arms for each study part are described here
 
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Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)
Part 3A (fed and fasted)
Part 3B (fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 1
(10mg/40mg)
ambrisentan and tadalafil 
FDC ) (10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 2
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg) fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 3
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
Section 1 - Protocol Synopsis Type and Number of Subjects
CHANGE FROM: 
A approximately of 20 healthy adult subjects will be randomized, to each study part, such 
that at least 16 evaluable subjects complete each part of the study
 
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CHANGE TO: 
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable 
subjects complete each part of the study.  Approximately 33 healthy subjects will be 
randomized to study Part 3A, such that at least 26 evaluable subjects complete part 
3A of the study. An additional 33 healthy subejcts will be randomised to Part 3B, 
such that at least 26 evaluable subjects complete part 3B of the study.
Section 1 - Protocol Synopsis Analysis
CHANGE FROM: 
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and Part 2, (ii) estimate the bioavailability of the formulation(s) of 
the FDC formulations, taken in Part 2, if used for food effect and  Part 3, in the fed state 
relative to the fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
CHANGE TO:
No formal hypothesis will be tested for Part 1 and Part 2 of the study. An estimation 
approach will be used to estimate the bioavailability of the FDC formulations 
relative to the monotherapies taken concurrently.  
For each primary pharmacokinetic endpoint, point estimates and corresponding 
90% confidence intervals will be constructed for the ratio of the geometric mean of 
the test treatment (FDC formulations) to the geometric mean of the reference 
treatment (co-administration of ambrisentan and tadalafil), (test)/ (reference).
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 
10 mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 
10 mg & tadalafil 40 mg taken concurrently (reference) in healthy subjects in both 
the fed and fasted states. The null hypothesis is that the true ratio of the geometric 
mean of the test treatment to the geometric mean of the reference treatment, 
(test)/(reference), for each primary PK endpoint, is either less than 0.80 or 
greater than 1.25.  The alternate hypothesis is that the true ratio of the test 
treatment geometric mean to the reference treatment geometric mean is at least 0.80 
and no greater than 1.25.  Symbolically, this is expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
 
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i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e., treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test 
(TOST) procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be 
used to test this set of hypotheses.  This is equivalent to requiring that a 90% 
interval for the true ratio of test to reference geometric means falls entirely within 
the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test the bioequivalence 
of a FDC of ambrisentan 5 mg + tadalafil 40 mg (test) and  of ambrisentan 5 mg + 
tadalafil 20 mg (test) relative to reference monotherapies of  ambrisentan 5 mg & 
tadalafil 40 mg and of ambrisentan 5 mg & tadalafil 20 mg respectively taken 
concurrently (reference) in healthy subjects in the fasted states. Therefore, the 
analysis approach for bioequivalence for Part 3A is  applicable to Part 3B.
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
Section 2 Introduction
CHANGE FROM
Ambrisentan (E.U. trade name: Volibris), an orally active endothelin receptor antagonists 
(ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first approved on 
15 June 2007 in the US and on 21 April 2008 in the European Union (EU) and is 
currently approved in over 50 countries. In the EU, ambrisentan is indicated for treatment 
of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional Class 
(FC) II to III, including use in combination treatment (Volibris, EMA SmPC, 2015 
Section 5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated 
with connective tissue disease
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (EMEA, 2010; Schuirmann, 1987;Adcirca, 2015)A recently completed 
study (Galiè, 2015a) has show that patients with PAH who started initial combination 
therapy with ambrisentan and tadalafil had a significantly lower risk of clinical-failure 
events compared to those that started with ambrisentan or tadalafil monotherapy. 
Ambrisentan has recently received EU approval (20 November 2015) for use in 
combination treatment with tadalafil (Volibris EMA SmPC, 2015, Section 5.1). 
 
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This pilot study will investigate the relative bioavailability of new fixed dose 
combinations (FDC) of ambrisentan and tadalafil, compared to the two monotherapies 
taken concurrently in healthy subjects
CHANGE TO
Ambrisentan (E.U. trade name: Volibris), is an orally active endothelin receptor 
antagonist (ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first 
approved on 15 June 2007 in the US and on 21 April 2008 in the European Union (EU) 
and is currently approved in over 50 countries. In the EU, ambrisentan is indicated for 
treatment of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional 
Class (FC) II to III, including use in combination treatment (Volibris EMA SmPC, 2015 
Section 5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated 
with connective tissue disease.
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (EMEA, 2010; Schuirmann, 1987;Adcirca, 2015).
A recently completed study (Galiè, 2015a) has shown that patients with PAH who started 
initial combination therapy with ambrisentan and tadalafil had a significantly lower risk 
of clinical-failure events compared to those that started with ambrisentan or tadalafil 
monotherapy. Ambrisentan has recently received EU approval (20 November 2015) for 
use in combination treatment with tadalafil (Volibris EMA SmPC, 2015, Section 5.1).
This pilot study will investigate the relative bioavailability of new fixed dose 
combinations (FDC) of ambrisentan and tadalafil, compared to the two monotherapies 
taken concurrently in healthy subjects
Section 2.1 Study Rationale
CHANGE FROM
This study is designed to select one, or more, FDCs of ambrisentan and tadalafil for 
further development and to provide pharmacokinetic data to enable a pivotal 
bioequivalence study (BE). Dependent on formulation work, the study will allow up to 8 
new FDCs to be compared with the reference of ambrisentan and tadalafil monotherapies 
taken concurrently. The study would also allow for up to 2 of the new formulations, that 
may be taken in to a BE study, to be tested for any effect on pharmacokinetics of the 
FDC in both fed and fasted state.
The formulation(s) to be taken forward into the BE study will be primarily chosen on the 
test/reference ratio for both AUC and Cmax for both components ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, between subject variability, 
tablet size, cost, ease of manufacture and stability would be considered.
 
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CHANGE TO
This study is designed initially to compare the relative bioavailability of a number of
fixed dose combinations (FDCs) of ambrisentan and tadalafil (Part 1 and 2) and 
consequently the bioequivalence of the FDC of different dose strengths (Part 3A and 
3B). Dependent on formulation work, the study will allow up to 9 new FDCs to be 
compared with the reference of ambrisentan and tadalafil monotherapies taken 
concurrently
The formulation(s) to be taken forward in Part 3A  will be primarily chosen based on the 
test/reference ratio for both AUC and Cmax for both components. Ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, tablet size, cost, ease of 
manufacture and stability would be considered. 
Part 3 will include evaluation of bioequivalence for 3 dose strengths of the FDC; 
10/40mg, 5/40mg and 5/20mg of ambrisentan and tadalafil.
Section 2.2 Brief Background
CHANGE FROM
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris EMA SmPC, 2015 Section 5.1). Efficacy has been 
shown in idiopathic PAH (IPAH) and in PAH associated with connective tissue 
disease.(Volibris, EMA SmPC,2015). Ambrisentan is an oral, once daily, propanoic acid-
based, ETA-selective Endothelin receptor antagonist (ERA) which targets the 
phospholipase-C-dependent endothelin pathway and which is known to play an essential 
role in mammalian cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (EMEA, 2010; Schuirmann, 
1987;Adcirca, 2015).Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) 
 
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inhibitor which targets the NO pathway. Through inhibition of PDE-5, tadalafil increases 
cytoplasmic cGMP concentrations in the smooth muscle cells and enhances NO-mediated 
vasodilatation of the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy subjects was performed to detect 
any significant PK interactions between tadalafil and ambrisentan when co-administered 
(Spence, 2009). From this study, it was concluded that there is no clinically significant 
PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when combined. 
Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
administered. . There were no SAE’s in the study. Three subjects withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one subject because of myalgia (severe), 
muscle fatigue and dizziness on tadalafil alone and one because of headache (severe) on 
the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild and is a 
listed event for ambrisentan. There were a total of 7 subjects with mild tachycardia. There 
were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. There were 3 events on 
the first day of tadalafil 40 mg given 3 days after ambrisentan 10 mg. There was one 
event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of ambrisentan 10 mg. 
Taken together these data suggest that in healthy subjects a mild tachycardia may result 
from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, EMA SmPC, 
2015;EMEA, 2010; Schuirmann, 1987;Adcirca, 2015).
The AMBITION clinical study (Galiè, 2015a and AMB112565 CSR 13Nov2014), which 
evaluated the time to first clinical failure event, a composite endpoint, shows a robust 
clinical benefit (50% hazard reduction) for PAH patients initiated on a combination of 
ambrisentan and tadalafil when compared to PAH patients initiated on either medication 
as monotherapy.  The safety profile of the combination arm was consistent with the 
known safety data of the individual study drugs and no safety signals specific to 
combination treatment were identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
 
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The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 
CHMP/EWP/240/95) (2008).
CHANGE TO
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris, EMA SmPC, 2015 Section 5.1). Efficacy has been 
shown in idiopathic PAH (IPAH) and in PAH associated with connective tissue disease
(Volibris, EMA SmPC, 2015). Ambrisentan is an oral, once daily, propanoic acid-based, 
ETA-selective Endothelin receptor antagonist (ERA) which targets the phospholipase-C-
dependent endothelin pathway and which is known to play an essential role in 
mammalian cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (EMEA, 2010; Schuirmann, 
1987;Adcirca, 2015). Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) 
inhibitor which targets the NO pathway. Through inhibition of PDE-5, tadalafil increases 
cytoplasmic cGMP concentrations in the smooth muscle cells and enhances NO-mediated 
vasodilatation of the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
 
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Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy participants was performed to 
detect any significant PK interactions between tadalafil and ambrisentan when co-
administered (Spence, 2009). From this study, it was concluded that there is no clinically 
significant PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when 
combined. Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
administered.  There were no SAE’s in the study. Three participants withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one participant because of myalgia 
(severe), muscle fatigue and dizziness on tadalafil alone and one because of headache 
(severe) on the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild 
and is a listed event for ambrisentan. There were a total of 7 participants with mild 
tachycardia. There were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. 
There were 3 events on the first day of tadalafil 40 mg given 3 days after ambrisentan 10 
mg. There was one event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of 
ambrisentan 10 mg. Taken together these data suggest that in healthy subjects a mild 
tachycardia may result from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, EMA SmPC, 
2015;EMEA, 2010; Schuirmann, 1987;Adcirca, 2015)The AMBITION clinical study 
(Galiè, 2015a and GSK document number 2014N193963_00), which evaluated the time 
to first clinical failure event, a composite endpoint, shows a robust clinical benefit (50% 
hazard reduction) for PAH patients initiated on a combination of ambrisentan and 
tadalafil when compared to PAH patients initiated on either medication as monotherapy.  
The safety profile of the combination arm was consistent with the known safety data of 
the individual study drugs and no safety signals specific to combination treatment were 
identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
 
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and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 2008).
Section 4.1 Overall Design
CHANGE FROM 
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects. See Figure 1 for study schematic.
Figure 2
Study Schematic
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
 
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There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose. 
The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast.
Subjects who had taken part in the study part preceding Part 3, and which provided the 
successful formulations for a BE study could also be invited to take part in this study 
part. However, a subject’s inclusion in more than one study part would be dependent on 
the subject not exceeding the maximum blood draw volume (500ml) for the study.
 
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CHANGE TO
This is a single centre, Phase 1, single dose, randomised, open label, crossover study in 
healthy volunteers with 3 study parts; Part 1 will include a 5 way cross-over, and 
Part 2 and 3 (A&B) will each include a 4 way cross-over. See Figure 1 for study 
schematic.
Figure 1
Study Schematic
Screening
≤ 31 days
Follow 
Up
Screening 
≤ 31 days from
Subjects 
Randomised to  
4 dose sessions 
(3 granulation 
formulations + 
reference)
Follow 
Up
Subjects 
randomised to 4 
dose sessions 
(FDC Fed & 
fasted + 
reference (fed 
&fasted))
Screening 
≤ 31 days
Follow 
Up
~4 Weeks
~4 Weeks
~4 Weeks
PART 1 (BA) n=20
PART 2 (BA) n=20
PART 3A (BE 10/40) n=33
Subjects 
Randomised to 
up to 5 dose 
sessions (4 FDC 
formulations 
and the 
reference)
Screening 
≤ 31 days
Subjects 
Randomised to 
4 dose sessions    
(2 FDC 
formulations 
(fasted) + 2 
reference fasted)
Follow 
Up
PART 3B (BE 5/40 & 5/20)
n=33
~4 Weeks
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in study Part 1 and 2. In 
study Part 3 there will be a minimum of 10 days between each dose.  A Follow-Up 
Visit should be completed within 7-14 days of completion of a subject’s last dose. 
The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have 5 dose sessions and will be used  to characterise the relative 
bioavailability, safety and tolerability of   four formulations of the fixed dose 
 
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combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
Part 2
This study part will have 4 dose sessions and will evaluate the bioavailability, safety and 
tolerability of  3 different granulation sizes for a single FDC (ambrisentan 10 mg + 
tadalafil 40 mg) compared to the reference of the 2 monotherapy components taken 
concurrently (ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted 
state.
Part 3A
Part 3A of the study is set to establish bioequivalence between the candidate FDC 
from Part 2. This study part will have 4 dose sessions and will assess the 
bioequivalence, in both the fed and fasted state, of the FDC against the reference  
ambrisentan 10 mg + tadalafil 40mg monotherapies in the fed and fasted state. The 
fed arms of this part will have a standard high fat breakfast (EMEA, 2010).
Part 3B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 
40mg and ambrisentan 5 mg + tadalafil 20mg) . This study part will have 4 dose 
sessions and will assess the bioequivalence of the two FDC dose strengths against the 
reference ambrisentan 5 mg + tadalafil 40mg monotherapies and ambrisentan 5 mg 
+ tadalafil 20mg monotherapies in the fasted state.
Section 4.2 Treatment and Duration
CHANGE FROM
The treatments for each study part of the study are listed in Table 1 All treatments are 
single dose. Subjects will be randomised to order of treatments in the parts of the study 
they are included in.
The study has 3 parts and ongoing analyse of pharmacokinetic data will be used to enable 
the formulations produced and tested in subsequent parts.
Part 1
Part 1 of the study will be utilised to look at up to 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
 
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Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2. 
If no successful formulations are identified from these criteria then the pharmacokinetic 
data would be used to enable further work to produce new FDC formulations for Part 2
Following completion of Part 1 there will be a pause prior to Part 2, so that up to a further 
4 FDC formulations could be produced and data included and approved in any required 
update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for up to 4 FDC formulations. These could be 
either successful formulation identified in Part 1 reformulated with the final API and/or 
new formulations using the final API. Pharmacokinetic data from Part 2 of the study will 
be analysed following completion of each treatment arm by subjects. This ongoing 
review of the pharmacokinetic data will enable rapid ongoing assessment of each FDC 
formulation and will be used to define any successful FDC formulation to be taken into 
Part 3 of this study and a pivotal BE study. If only two formulations are evaluated in Part 
2 then the food effect may be added to Part 2 and Part 3 will not be required. Success will 
be defined with the same criteria as those in Part 1.
Part 3
Part 3 of the study will provide data for up to 2 successful formulations to be progressed 
to a pivotal BE study, in both the fed and fasted state.
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil FDC5 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil FDC6 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil FDC7 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC4 (10mg/40mg)
ambrisentan and tadalafil FDC8 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. Subjects may be included in Part 1, Part 2 or Part 3 of the study. However, if Part 2 
of the study identifies any successful formulations to take into Part 3 then subjects from 
Part 2 may be invited to join Part 3.
The minimum and maximum duration for a subject on this study is described in Figure 2. 
This table assumes all 5 dose sessions are used for Part 1 and Part 2 and either 2 or 4 dose 
sessions are used in Part 3. However, this is an approximation as fewer sessions may be 
required in Part 2 and/or the washout between sessions of 7 days in each part may be 
longer, due to logistical reasons.
 
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Table 2
Study Duration
Study Options
Screen
Study 
Part 
1or 2
Pause
Study 
part 3
Follow-
Up Visit
Total 
Duration 
on Study
(1-31 
days)
(31 
days)
(7-28 
days)
(10-24 
days)
(7-14 
days)
Subjects 
included 
in only 
Part 1 or 
2
Min
1
31
7
39
Max
31
31
14
76
Subjects 
included 
in Part 
2&3
Min
1
31
7
10
7
56
Max
31
31
28
24
14
128
Subjects 
included 
in only 
Part 3
Min
1
10
7
18
Max
31
24
14
69
A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
CHANGE TO
The treatments for each study part of the study are listed in Table 1. All treatments are 
single dose. Subjects will be randomised to the order of treatments in the parts of the 
study they are included in.
The study has 3 parts and ongoing analysis of pharmacokinetic data will be used to 
determine the formulations produced and tested in subsequent parts.
Part 1
Part 1 of the study will be utilised to evaluate 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
 
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FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2
and 3. 
Following completion of Part 1 there will be a pause prior to Part 2, so that different 
granulation forms of a single FDC can be produced and data included and approved in 
any required update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for 3 granulation size forms of a single FDC 
formulation selected from Part 1. These will be based on a single successful 
formulation identified in Part 1 reformulated with the final active pharmaceutical 
ingredient (API). Success will be defined with the same criteria as those in Part 1.
Part 3 (A and B)
The BE part of the study will investigate the bioequivalence of the FDC ambrisentan 
10 mg + tadalafil 40 mg in part 3A and the bioequivalence of the FDC ambrisentan 
5 mg + tadalafil 40 mg and ambrisentan 5 mg + tadalafil 20 mg in Part 3B.  
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted) 
Part 3A (fed and fasted)
Part 3B (fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 1 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
(10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and tadalafil 
FDCgranulation 2
(10mg/40mg)
ambrisentan and 
tadalafil FDC
(10mg/40mg) fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 3
(10mg/40mg)
ambrisentan and 
tadalafil monotherapies 
taken concurrently
(10mg/40mg), fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently
(10mg/40mg)
ambrisentan and 
tadalafil monotherapies 
taken concurrently
(10mg/40mg), fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
The duration for a subject on this study is described in Figure 2. This figure assumes all 5 
dose sessions are used for Part 1. However, this is an approximation as minimum of 7 
days is expected between doses in Parts 1 and 2 and a minimum of 10 days in Part 3.
 
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Figure 2
Study Duration
Part 1 can last up to 73 days. 
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
Dose 
Period 5 
start 
7 
days
7 
days
7 
days
7 
days
7 to 
14 
days
Follow-up
 
 
 
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Part 2  will last approximately 9 weeks
Part 3A and B will last approximately 11 weeks
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
7 days
7 days
7 days
7 to 14  
days
Follow-up
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
10 
days
10  
days
10 
days
7 to 14  
days
Follow-up
 
 
 
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A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
Section 4.3 Type and Number of Subjects
CHANGE FROM
A approximately of 20 healthy adult subjects will be randomized, to each study part, such 
that at least 16 evaluable subjects complete each part of the study.
The study is not powered. However, the number of subjects will be sufficient for the 
objectives of this study and this is detailed further in Section 9.2 Sample Size 
Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
CHANGE TO
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable subjects 
complete each part of the study.  Approximately 33 healthy subjects will be 
randomized to study Part 3A, such that at least 26 evaluable subjects complete part 
3A of the study. An additional 33 healthy subjects will be randomised to Part 3B, 
such that at least 26 evaluable subjects complete part 3B of the study.
The estimation of the number of subjects for each study part is detailed further in 
Section 9.2 Sample Size Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
Section 4.4 Design Justification
CHANGE FROM
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations to take into a pivotal BE study and to also provide pharmacokinetic data to 
enable the pivotal BE study to be powered sufficiently.
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
 
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The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, or is there any need to blind study treatment. The Inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study is in three parts to enable PK data from Part 1 to enable formulation 
development for Part 2. Part 3 providing an opportunity to assess food effect in any FDC 
formulations suitable for development and inclusion in the pivotal bioequivalence study. 
The interdependency of the 3 study parts is described in detail in Section 4.2. 
CHANGE TO
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, nor is there any need to blind study treatment. The inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study contains three parts: Parts 1 and 2 will provide PK data for different 
candidate FDC formulations and will enable the assessment of bioequivalence of the 
FDC versus the combination of the individual reference products in Part 3. The 
interdependency of the 3 study parts is described in detail in Section 4.2. 
Section 4.5 Dose Justification
CHANGE FROM
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of, 40mg for tadalafil and 10mg for ambrisentan are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
be tolerated in healthy subjects.
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimum washout of 7 days, 
between doses, is also sufficient for clearance of previous dose.
 
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CHANGE TO
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of 40mg for tadalafil and 10mg for ambrisentan, are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
be tolerated in healthy subjects. Within Part 3 of the study all dosage forms of the 
FDC that are proposed for marketing authorisation will be tested for bioequivalence 
versus the appropriate doses of reference products.  These doses are required in 
clinical practice to allow safe up- and down-titration of the FDC in patients with 
PAH.  
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimumof 10 days, between 
doses, is also sufficient for clearance of the previous dose during the assessmet of 
bioequivalence.
Section 4.6.2 Benefit Assessment
CHANGE FROM
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation for a 
fixed dose combination of ambrisentan plus tadalafil to use as first line therapy in PAH 
patients.
CHANGE TO
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation and to 
demonstrate bioequivalence versus reference products for a fixed dose combination of 
ambrisentan plus tadalafil to use as first line therapy in PAH patients.
Section 4.6.3 Overall Benefit: Risk Conclusion
CHANGE FROM
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 3. So, the 
risk: benefit is appropriate for this study and to enable development of this investigational 
product. The overall benefit: risk therefore remains positive.
 
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CHANGE TO
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 3. In 
addition review of safety information from Part 1 of the study is consistent with this 
position. So, the benefit:risk is appropriate for this study and to enable development of 
this investigational product. The overall benefit: risk therefore remains positive.
Section 6.1 Investigational Product and Other Study Treatment
CHANGE FROM
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Part 1 of the study are shown in Table 4, this will be 
amended once treatments for Part 2 and Part 3 are confirmed.
 
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Table 4
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS*
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS*
*
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 95 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 233 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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CHANGE TO
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Parts 1, 2 and 3 of the study are shown in Table 4, 
Table 5 and Table 6.
 
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Table 4
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 90 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.45
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Address of 
reference 
manufacturer
NA
NA
NA
NA
Glaxo Operations UK 
Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 5 
Study Treatment for Part 2
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 granulation 
1 (10mg/40mg)
FDC-G2 granulation 2 
(10mg/40mg)
FDC-G3 granulation 
3 (10mg/40mg)
Each tablet contains 10 
mg of ambrisentan, 
approximately 90 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg of 
lecithin (soya) (E322) and 
approximately 0.45 mg of 
Allura red AC Aluminium 
Lake 
Each film-coated 
tablet contains 20 mg 
tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage level(s):
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 10 
mg of ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration 
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC
FDC
FDC
Reference
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations UK Ltd 
(trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la Industria 
30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 6
Study Treatment for Part 3
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-
Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil -
Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 or 2 or3
(10mg/40mg)
FDC  (5mg/40mg)
FDC  (5mg/20mg)
Each tablet contains 
10 mg of 
ambrisentan, 
approximately 90 mg 
of lactose (as 
monohydrate), 
approximately 0.25 
mg of lecithin (soya) 
(E322) and 
approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each tablet 
contains 5 mg of 
ambrisentan, 
approximately 95 
mg of lactose (as 
monohydrate), 
approximately 
0.25 mg of 
lecithin (soya) 
(E322) and 
approximately 
0.11 mg of Allura 
Red AC 
Aluminium Lake
Each film-coated 
tablet contains 20 
mg tadalafil, 245 
mg lactose (as 
monohydrate).
Dosage form:
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 5 mg 
of ambrisentan.
Each tablet contains 
20 mg tadalafil and 5 
mg of ambrisentan.
Each tablet contains 
10 mg of 
ambrisentan.
Each tablet 
contains 5 mg of 
ambrisentan.
Each tablet 
contains 20 mg 
tadalafil.
Route of 
Administration 
Oral
Oral
Oral
Oral
Oral
Oral
 
 
 
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Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-
Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil -
Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations 
UK Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Glaxo Operations 
UK Ltd (trading 
as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 
Alcobendas
Madrid
Spain
 
 
 
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6.1.1 Retention Samples
GSK will send five times (5x) the drug product necessary for pivotal bioequivalency 
evaluation in Part 3 of the study for analytical testing for all tablet strengths used in 
the study. The retain samples are in addition to the supply of drug product 
sufficient to complete the study. Retention samples will be sent to the clinical CRO 
in the same bulk container. To ensure that reserve samples are representative of the 
same batches provided for the clinical study the clinical site will randomly select the 
supplies for the study from the supply received. The clinical site must retain enough 
reserve samples to permit FDA to perform five times all of the release tests required 
in the application.
Section 6.3 Treatment Assignment
CHANGE FROM
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software.
The treatments are denoted as F1 to F4 for Part 1, F5 to F8 for Part 2 and R for reference 
treatment for both parts; the selected formulations for Part 3 are denoted as X and Y.  The 
treatment key for Parts 1, 2 and 3 are described in Table 6
Table 6 
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 2
F5
ambrisentan and tadalafil FDC5 (10mg/40mg)
F6
ambrisentan and tadalafil FDC6 (10mg/40mg)
F7
ambrisentan and tadalafil FDC7 (10mg/40mg)
F8
ambrisentan and tadalafil FDC8 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3
 
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Treatment
Description
X1
ambrisentan and tadalafil FDCX1 (10mg/40mg), fed
X2
ambrisentan and tadalafil FDCX1 (10mg/40mg) fasted
Y1
ambrisentan and tadalafil FDCX2 (10mg/40mg), fed
Y2
ambrisentan and tadalafil FDCX2 (10mg/40mg), fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 7. Not all possible sequences are 
included here. Such as, if only one formulation taken into Part 2 or 3, or Part 2 is used for 
only one or two FDC formulations, fed and fasted. Additional treatment sequences will 
be created based on the Latin Squares for Williams Designs, as required
Table 7 
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1 F2 R F3 F4
F2 F3 F1 F4 R
F3 F4 F2 R F1
F4 R F3 F1 F2
R F1 F4 F2 F3
F4 F3 R F2 F1
R F4 F1 F3 F2
F1 R F2 F4 F3
F2 F1 F3 R F4
F3 F2 F4 F1 R
1:1:1:1:1:1:1:1:1:1
Part 2
5
F5 F6 R F7 F8
F6 F7 F5 F8 R
F7 F8 F6 R F5
F8 R F7 F5 F6
R F5 F8 F6 F7
F8 F7 R F6 F5
R F8 F5 F7 F6
F5 R F6 F8 F7
F6 F5 F7 R F8
F7 F6 F8 F5 R
1:1:1:1:1:1:1:1:1:1
4
F5 F6 R F7
F6 F7 F5 R
F7 R F6 F5
R F5 F7 F6
1:1:1:1
3
F5 F6 R
R F6 F5
F6 R F5
F1 R F6
R F5 F6
F6 F5 R
3:3:3:3:3:3 + 2 random
2
F5 R 
1:1
 
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Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
R F5
Part 3
4
X1 X2 Y2 Y1
X2 Y1 X1 Y2
Y1 Y2  X2 X1
Y2  X1 Y1 X2 
1:1:1:1
Part 2 and 
Part 3 
merged 
together
5 (Two FDCs selected from 
Part 1)
X1 X2 R Y1 Y2
X2 Y1 X1 Y2 R
Y1 Y2 X2 R X1
Y2 R Y1 X1 X2
R X1 Y2 X2 Y1
Y2 Y1 R X2 X1
R Y2 X1 Y1 X2
X1 R X2 Y2 Y1
X2 X1 Y1 R Y2
Y1 X2 Y2 X1 R
1:1:1:1:1:1:1:1:1:1
3 (One FDC selected from 
Part 1)
X1 X2 R
R X2 X1
X2 R X1
F1 R X2
R X1 X2
X2 X1 R
3:3:3:3:3:3 + 2 random
CHANGE TO
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software. The treatments are 
denoted as F1 to F4 for Part 1, F5 to F8 for Part 2 and R for reference treatment for both 
parts; the selected formulations for Part 3 are denoted as X and Y
The treatment key for Parts 1, 2 and 3 are described in Table 6.
Table 6
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
 
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Treatment
Description
Part 2
FG1
ambrisentan and tadalafil FDC-G1 (10mg/40mg) granulation size 1
FG2
ambrisentan and tadalafil FDC-G2 (10mg/40mg) granulation size 2
FG3
ambrisentan and tadalafil FDC-G3 (10mg/40mg) granulation size 3
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3A
X1
ambrisentan and tadalafil FDC-G1 or 2 or 3  (10mg/40mg) fed
X2
ambrisentan and tadalafil FDC G1 or 2 or 3 (10mg/40mg) fasted
R1
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fed
R2
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fasted
Part 3B
Y1
ambrisentan and tadalafil FDC (5mg/40mg), fasted
Y2
ambrisentan and tadalafil FDC (5mg/20mg), fasted
R3
ambrisentan and tadalafil monotherapies taken concurrently 
(5mg/40mg), fasted
R4
ambrisentan and tadalafil monotherapies taken concurrently 
(5mg/20mg), fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 7. Not all possible sequences are 
included here. Such as, if only one formulation taken into Part 2 or 3, or Part 2 is used for 
only one or two FDC formulations, fed and fasted Additional treatment sequences may be 
created based using the Latin Squares for Williams Designs if needed.
Table 7
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1/ F2/ R/ F3/ F4
F2/ F3/ F1/ F4/ R
F3/ F4/ F2/ R/ F1
F4/ R/ F3/ F1/ F2
R/ F1/ F4/ F2/ F3
F4/ F3/ R/ F2/ F1
R/ F4/ F1/ F3/ F2
F1/ R/ F2/ F4/ F3
F2/ F1/ F3/ R/ F4
F3/ F2/ F4/ F1/ R
1:1:1:1:1:1:1:1:1:1
 
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Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 2
4
FG1/ FG2/ R/ FG3
FG2/ FG3/ FG1/ R
FG3/ R/ FG2/ FG1
R/ FG1/ FG3/ FG2
1:1:1:1
Part 3A
4
X1/ R1/ R2/ X2
R1/ X2/ X1/ R2
X2/ R2/ R1/ X1
R2/ X1/ X2/ R1
1:1:1:1
Part 3B
4
Y1/ R3/ R4/ Y2
R3/ Y2/ Y1/ R4
Y2/ R4/ R3/ Y1
R4/ Y1/ Y2/ R3
1:1:1:1
Section 9.1 Hypotheses
CHANGE FROM
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and if used, Study part 2, (ii) estimate the bioavailability of the 
formulation(s) of the FDC formulations, taken in to Part 3, in the fed state relative to the 
fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
CHANGE TO
No formal hypothesis will be tested for study Part 1 and Part 2. An estimation 
approach  will be used to (i) estimate the bioavailability of the FDC formulations relative 
to the monotherapies taken concurrently, (ii) for each primary pharmacokinetic 
endpoint, point estimates and corresponding 90% confidence intervals will be 
constructed for the ratio of the geometric mean of the test treatment (FDC 
formulations) to the geometric mean of the reference treatment (co-administration 
of ambrisentan and tadalafil), (test)/ (reference).  The objective of Part 2 is to 
assess whether differences in granulation size impact the pharmacokinetics of 
ambrisentan and tadalafil; the estimation approach for bioavailability is therefore 
applicable to Part 2.
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 
10 mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 
10 mg & tadalafil 40 mg taken concurrently (reference) in healthy subjects in both 
the fed and fasted states. The null hypothesis is that the true ratio of the geometric 
 
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mean of the test treatment to the geometric mean of the reference treatment, 
(test)/(reference), for each primary PK endpoint, is either less than 0.80 or 
greater than 1.25.  The alternate hypothesis is that the true ratio of the test 
treatment geometric mean to the reference treatment geometric mean is at least 0.80 
and no greater than 1.25.  Symbolically, this is expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e. , treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test 
(TOST) procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be 
used to test this set of hypotheses.  This is equivalent to requiring that a 90% 
interval for the true ratio of test to reference geometric means falls entirely within 
the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test (1) the 
bioequivalence of a FDC of ambrisentan 5 mg + tadalafil 40 mg (test) relative to 
reference monotherapies of ambrisentan 5 mg & tadalafil 40 mg taken concurrently 
(reference) and (2) the bioequivalence of a FDC of ambrisentan 5 mg + tadalafil 20 
mg (test) relative to reference monotherapies of ambrisentan 5 mg & tadalafil 20 mg 
taken concurrently (reference), in healthy subjects in the fasted states. Therefore, 
the analysis approach for bioequivalence for Part 3A is applicable to Part 3B.
Section 9.2 Sample Size Considerations
CHANGE FROM:
No formal sample size calculation has been performed. Sample sizes chosen for each 
study part in this study is considered sufficient for exploratory analysis based on PK 
variability data from previous studies
CHANGE TO
No formal sample size calculation has been performed. Sample sizes chosen for each 
study part in this study is considered sufficient for exploratory analysis based on PK 
variability data from previous studies
Section 9.2.1 Sample Size Assumptions
 
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CHANGE FROM
The sample size assumptions are based on previously reported estimates of within subject 
CV for AUC(0-∞) and Cmax for ambrisentan (GS-US-300-0112, 2008) and tadalafil 
(Forgue, 2005). Table 9 summarizes the estimates of within subject CV for the primary 
endpoints AUC (0-∞) and Cmax.
Table9 
Estimates of within subject CV for the primary end points AUC (0-∞)
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
CHANGE TO
For Part 1 and Part 2 of the study, the sample size assumptions are based on 
previously reported estimates of within subject CV for AUC(0-∞) and Cmax for 
ambrisentan (GS-US-300-0112, 2008) and tadalafil (Forgue, 2005). Table 9 summarizes 
the estimates of within subject CV for the primary endpoints AUC (0-∞) and Cmax.
Table 9 
Estimates of within subject CV for the primary end points AUC (0-∞) 
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
For Part 3A of the study, Table 10 summarized the observed estimates of between 
and within subject CV and observed ratio of geometric means for the primary 
endpoints AUC(0-t) and Cmax from Part 1 of the study.
 
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Table 10
Estimates of within subject CV for the primary end points AUC 
(0-∞) and Cmax for ambrisentan and tadalafil from part 1 of the study
CVw: within subject CV
ambrisentan 
tadalafil 
Cmax
22.42%
14.53%
AUC (0-∞)
8.12%
10.32%
The largest of the within subject CV estimates 22.42% translates to a standard 
deviation (SD) of 0.221 on the natural log scale.  Based on this SD, a sample size of 
23 statistically evaluable subjects will have 90% power to demonstrate 
bioequivalence. This calculation assumes:

a true ratio of 1,

the within-subject variability from the current study will not be larger than 
that used in the sample size calculations,

data are log-normally distributed, and each t-test is made at the 5% level.
For Part 3B of the study, same assumptions are adopted as Part 3A. Thus, a sample 
size of 23 statistically evaluable subjects will have 90% power to demonstrate 
bioequivalence.  However, if the within subject CVs obtained after Part 2 are 
significantly higher than the ones in Part 1 (shown in Table 10), a sample size re-
esimation may be conducted to adjust the sample size for Part 3A and 3B to ensure 
the study maintain enough power to demonstrate bioequivalence.
Section 9.2.2 Sample Size Sensitivity
CHANGE FROM
Assuming the within subject CV differs from 22% and the number of statistically 
evaluable subjects is lower than 20, then Table 11 shows the precision (the half width of 
the 90% confidence interval) for a range of evaluable subjects.
 
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Table 11
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
CHANGE TO
For Part 1 and Part 2, assuming the within subject CV differs from 22% and the 
number of statistically evaluable subjects is lower than 20, then Table 11 shows the 
precision (the half width of the 90% confidence interval) for a range of evaluable 
subjects.
Table 11
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
For Part 3, based on the observed data from previous studies, the ratio of geometric 
mean for ambrisentan Cmax is unlikely to be 1. The effects on the power of 
declaring bioequivalence in the face of a shift in the expected ratio of the geometric 
means were examined.
 
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If, under all other assumptions outlined above, the actual ratio of geometric means 
for ambrisentan Cmax is 0.97, then 26 evaluable subjects are needed to have 90% 
power to conclude bioequivalence.
With 26 evaluable subjects, the current study design has at least 90% power to 
conclude bioequivalance (with both Cmax and AUC for ambrisentan and tadalafil).
Section 9.2.3 Sample Size Re-estimation or Adjustment
CHANGE FROM
No sample size re-estimation will be performed.
CHANGE TO
No sample size re-estimation will be performed.  However, if the estimation of the 
within subject CV from Part 2 of the study is much larger than the within subject 
CV from Part 1 of the study, the sample size for Part3 may be re-evaluated.
Section 9.3 Data Analysis Considerations
CHANGE FROM
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
sampling times recorded during the study.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-
life (t1/2).  
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model.
CHANGE TO
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
 
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sampling times recorded during the study although supplementary analayis will be 
available based on the nominal times.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-life 
(t1/2).  
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model
9.3.1
Analysis Populations
9.3.1.1.
Safety Population
All subjects enrolled into the study who have received at least one dose of investigational 
product will be included in the Safety Population.
9.3.1.2.
Pharmacokinetic Concentration Population
The PK Concentration Population will include all subjects for whom at least one PK 
sample was obtained and analyzed.
9.3.1.3.
Pharmacokinetic Parameter Population
For each PK parameter, the PK Parameter Population will include all subjects who 
provide PK parameter data.
Section 9.3.2 Interim Analysis
CHANGE FROM
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in order to direct development of fixed dose formulations for subsequent 
study parts and the planned BE study.
CHANGE TO
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in Part 1 and Part 2 of the study in order to direct development of the 
FDC for later parts of the study.  Headline results based on statistical analysis using 
preliminary pharmacokinetic data with nominal time may be produced when 80% 
and 100% subjects complete Part 1 and Part 2 of the study to assist development of 
 
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FDCs.  Treatment and period information from the crossover design may be used in 
the analysis; sequence information may also be included depending on availability.
Section 9.4.1 Pharmacokinetic Analyses
CHANGE FROM
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D. 
Following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using a mixed effects model with fixed effect 
terms for period and treatment. Subject will be treated as a random effect in the model. 
Point estimates and their associated 90% confidence intervals will be constructed for the 
differences, F1 – R, F2 – R, F3 – R, F4 – R for Part 1 and similarly for Part 2 (as needed); 
and X1-X2, Y1-Y2 for Part 3. The point estimates and their associated 90% confidence 
intervals will then be back-transformed to provide point estimates and 90% confidence 
intervals for the ratios, F1:R, F2-R:F3-R, F4-R; and X1:X2, Y1:Y2.
Tmax of FDC formulations will be analyzed with the non-parametric Wilcoxon Matched 
Pairs Method to compute point estimate and associated 90% confidence intervals for the 
median difference, F1 R, F2 R, F3 R, F4 R and X1:X2, Y1:Y2.
CHANGE TO
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D.
Data will be listed and summarized according to GlaxoSmithKline reporting 
standards, where applicable.  Listings will be sorted by subject, period, noting 
treatment.  Summaries will be presented by treatment.  Unless stated otherwise, 
descriptive summaries will include n, mean, standard deviation (SD), coefficient of 
variation (CV), median, minimum, and maximum for continuous variables, n and 
percent for categorical variables and geometric mean, 95% confidence interval (CI), 
and the between-subject CV (CVb) based on geometric mean for the log-
transformed PK parameters.
Version 9.3 (or higher) of the SAS system will be used for statistical analysis of the 
data as well as to generate tables, figures, and listings.
 
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Any deviation(s) from the original analyses planned in the protocol will be reported 
in the Reporting and Analysis Plan (RAP) and/or in the Clinical Pharmacology 
Study Report.
The PK parameters, AUC(0-), AUC(0-t), Cmax, and t½ will be transformed using 
natural logarithms. Missing PK parameters will not be imputed.
All data from withdrawn subjects will be listed.
No adjustment for multiple tests or comparisons is planned.
Actual visit day will be used for safety assessments. For the assessment of 
bioequivalence, both actual time of sampling, and nominal (planned) sampling time, 
will be used to estimate PK parameters at the end of the study.  PK parameters 
estimation based on nominal sampling time will be used for generating headline 
results.Following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using a mixed effects model with fixed effect 
terms for period, sequence and treatment, and subject as random effect term (i.e. 
ANOVA method using period, sequence and treatment as fixed effect, and subject as 
random effect) Point estimates and their associated 90% confidence intervals will be
calculated for the differences: F1 - R, F2 - R, F3 -R, F4 - R for Part 1; FG1-R, FG2-R, 
FG3-R for Part 2; X1-R1, X2-R2 for Part 3A; and Y-R3 for Part 3B. The point 
estimates and their associated 90% confidence intervals will then be back-transformed to 
provide point estimates and 90% confidence intervals for the ratios, F1:R, F2:R, F3:R, 
F4:R; for Part 1; FG1:R, FG2:R, FG3:R for Part 2; X1:R1,X2:R2 for Part3A; and 
Y:R3 for Part 3B.
Section 11 REFERENCES
ADDITIONS
EMEA guidelines on the investigation of bioequivalence. CPMP/EWP/QWP/1401/98 
Rev. 1/ Corr **. 2010
Schuirmann DJ. A comparison of the two one-sided tests procedure and the power 
approach for assessing the equivalence of average bioavailability, J Pharmacokin. 
Biopharm. 1987:15:657–680. 
Section 12.4 Appendix 4 High Fat Meal Content
CHANGE FROM

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
 
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The standard high-fat meal will be the meal suggested by the US FDA in their 2002 draft 
guidance on food-effect bioavailability and bioequivalence studies. Approximately 50% 
of the caloric content of the meal is from fat and the meal is high in calories 
(approximately 1000 calories). Substitutions in this test meal can be made as long as the 
meal provides a similar amount of calories from protein, carbohydrate, and fat and has 
comparable meal volume and viscosity.
US Department of Health and Human Services.  Guidance for Industry. Food-Effect 
Bioavailability and Fed Bioequivalence Studies.  Guidance. Center for Drug Evaluation 
and Research (CDER). Food and Drug Administration; 2002.
CHANGE TO

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
The standard high-fat meal will be the meal suggested by the EMEA guidelines on the 
investigation of bioequivalence 2010. The meal should be a high-fat (approximately
50 percent of total caloric content of the meal) and high-calorie (approximately 800 to
1000 kcal) meal. This test meal should derive approximately 150, 250, and 500-600 
kcal from protein, carbohydrate, and fat, respectively(EMEA, 2010).
 
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12.5.3.
Protocol changes for Amendment 3 (25-MAY-2017) from 
Amendment 3 (8-Feb-2017)
Amendment 3 Summary of Changes
The SmPC and PIL for tadalafil (Adcirca) have recently been updated to reflect the risk 
of a sudden decrease or loss of vision or hearing. The risk of visual defects and Non-
Arteritic Anterior Ischemic Optic Neuropathy (NAION) have been reported with the use 
of tadalafil (and other PDE5 inhibitors) for the management of erectile dysfunction 
though maybe relevant other patients exposed to tadalafil. Cases of sudden hearing loss 
have been reported with the use of tadalafil. Though other risk factors for hearing loss 
were present, this may be relevant to other patients exposed to tadalafil. Therefore, the 
protocol (and patient PIL) have been updated to reflect these changes. The GSK medical 
monitor will also change. Specific sections relating to Data Analyses considerations and 
PK Analysis have been updated to reflect EMA guidance.
List of Specific Changes
Section 4.6 Benefit: Risk Assessment
CHANGE FROM
Summaries of findings from clinical studies conducted with both the Investigational 
Product taken as monotherapies and in combination can be found in: 

Ambrisentan IB (Volibris IB, 2015)

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015) 

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010; Schuirmann, 1987;Adcirca, 2015)
CHANGE TO
Summaries of findings from clinical studies conducted with both the Investigational 
Product taken as monotherapies and in combination can be found in: 

Ambrisentan IB (Volibris IB, 2015)

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015) 

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010; Schuirmann, 1987;Adcirca, 2017)
Section 4.6.1 Risk Assessment
Table 2
Risks of Clinical Significance and Mitigation Strategy
CHANGE FROM:
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
Investigational Product: GSK1325760 (ambrisentan-Volibris)

Teratogenicity

Anaemia (decreased 
haemoglobin, decreased 
haematocrit)

Hypersensitivity reactions 
(e.g. angioedema, rash, 
pruritus)

Headache (including sinus 
headache, migraine)

Dizziness

Cardiac failure

Palpitation

Hypotension

Flushing

Epistaxis

Dyspnoea

Upper respiratory  
congestion, sinusitis, 
nasopharyngitis, rhinitis

Abdominal pain

Constipation

Nausea, vomiting,
diarrhoea

Hepatic transaminases 
increased

Peripheral oedema, fluid 
retention

Chest pain/discomfort

Asthenia and fatigue
Ambrisentan IB (Volibris IB, 
2015)
Volibris (Ambrisentan) EMA 
SmPC (Volibris EMA SmPC, 
2015): 
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision. 

Exclusion of women of 
childbearing potential 
(teratogenicity)

Laboratory assessments 
per protocol

Physical assessment per 
protocol

Routine vital signs per 
protocol

Subjects remain in the 
clinical unit, under medical 
supervision, for all doses 
and until completion of 
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GF196960 (tadalafil - Adcirca)

Hypersensitivity reactions

Headache

Syncope,

Migraine

Blurred vision

Palpitations

Nasopharyngitis (including 
nasal congestion, sinus 
congestion and rhinitis)

Epistaxis

Nausea,

Dyspepsia (including 
Adcirca (Tadalafil) EMA SmPC 
(EMEA, 2010; Schuirmann, 
1987;Adcirca, 2015)
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision.

Routine vital signs per 
protocol

Physical assessment per 
protocol

Subjects remain in the 
clinical unit for all doses, 
under medical supervision 
and until completion of 
safety assessments at 
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
abdominal pain/discomfort)

Vomiting,

Gastroesophageal reflux

Rash

Myalgia,

Back pain

Pain in extremity (including 
limb discomfort)

Increased uterine bleeding

Facial oedema,

Chest pain
48hrs post dose.

Single doses used in the 
study
Investigational Product: GSK3380154 (ambrisentan-tadalafil-FDC)
The risks for the combination 
are the same as the 2 
monotherapies; no extra risks 
have been identified for the 
combination.
As for GSK1325760 and 
GF196960 above
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
Investigational Product: GSK1325760 (ambrisentan-Volibris)

Teratogenicity

Anaemia (decreased 
haemoglobin, decreased 
haematocrit)

Hypersensitivity reactions 
(e.g. angioedema, rash, 
pruritus)

Headache (including sinus 
headache, migraine)

Dizziness

Cardiac failure

Palpitation

Hypotension

Flushing

Epistaxis

Dyspnoea

Upper respiratory  
congestion, sinusitis, 
nasopharyngitis, rhinitis

Abdominal pain

Constipation

Nausea, vomiting, 
diarrhoea

Hepatic transaminases 
increased

Peripheral oedema, fluid 
retention

Chest pain/discomfort

Asthenia and fatigue
Ambrisentan IB (Volibris IB, 
2015)
Volibris (Ambrisentan) EMA 
SmPC (Volibris EMA SmPC, 
2015): 
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision. 

Exclusion of women of 
childbearing potential 
(teratogenicity)

Laboratory assessments 
per protocol

Physical assessment per 
protocol

Routine vital signs per 
protocol

Subjects remain in the 
clinical unit, under medical 
supervision, for all doses 
and until completion of 
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GF196960 (tadalafil - Adcirca)

Hypersensitivity reactions

Headache

Syncope,

Migraine

Blurred vision

Sudden decrease or loss 
of vision (Non-Arteritic 
Anterior Ischemic Optic 
Neuropathy (NAION))

Palpitations

Nasopharyngitis (including 
nasal congestion, sinus 
Adcirca (Tadalafil) EMA SmPC 
(EMEA, 2010; Schuirmann, 
1987;Adcirca, 2017)
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision.

Routine vital signs per 
protocol

Physical assessment per 
protocol

Subjects remain in the 
clinical unit for all doses, 
under medical supervision 
and until completion of 
safety assessments at 
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
congestion and rhinitis)

Epistaxis

Nausea,

Dyspepsia (including 
abdominal pain/discomfort)

Vomiting,

Gastroesophageal reflux

Rash

Myalgia,

Back pain

Pain in extremity (including 
limb discomfort)

Increased uterine bleeding

Facial oedema,

Chest pain

Sudden decrease or loss 
of hearing
48hrs post dose.

Single doses used in the 
study
Investigational Product: GSK3380154 (ambrisentan-tadalafil-FDC)
The risks for the combination 
are the same as the 2 
monotherapies; no extra risks 
have been identified for the 
combination.
As for GSK1325760 and 
GF196960 above
Section 5 Selection of Study Population and Withdrawal Criteria
Rationale for change: as above.
CHANGE FROM:
Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in:

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015)

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010; Schuirmann, 1987;Adcirca, 
2015)
CHANGE TO:
 
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Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in:

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015)

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010; Schuirmann, 1987;Adcirca, 
2017)
Section 9.3 Data Analysis Consideration
Rationale for change: to reflect updated guidelines
CHANGE FROM :
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model
CHANGE TO :
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in fixed effect ANOVA model and mixed effects 
model
Section 9.4.1 Pharmacokinetic Analysis
Rationale for change: to reflect updated guidelines.
CHANGE FROM :
Actual visit day will be used for safety assessments. For the assessment of 
bioequivalence, both actual time of sampling, and nominal (planned) sampling time, will 
be used to estimate PK parameters at the end of the study. PK parameters estimation 
based on nominal sampling time will be used for generating headline results.Following 
loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations and reference will 
be separately analyzed using a mixed effects model with fixed effect terms for period, 
sequence and treatment, and subject as random effect term (i.e. ANOVA method using 
period, sequence and treatment as fixed effect, and subject as random effect) Point 
estimates and their associated 90% confidence intervals will be calculated for the 
differences: F1 - R, F2 - R, F3 -R, F4 - R for Part 1; FG1-R, FG2-R, FG3-R for Part 2; 
X1-R1, X2-R2 for Part 3A; and Y-R3 for Part 3B. The point estimates and their 
associated 90% confidence intervals will then be back-transformed to provide point 
estimates and 90% confidence intervals for the ratios, F1:R, F2:R, F3:R, F4:R; for Part 1; 
FG1:R, FG2:R, FG3:R for Part 2; X1:R1,X2:R2 for Part3A; and Y:R3 for Part 3B.
CHANGE TO :
 
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Actual visit day will be used for safety assessments. For the assessment of 
bioequivalence, both actual time of sampling, and nominal (planned) sampling time, will 
be used to estimate PK parameters at the end of the study. PK parameters estimation 
based on nominal sampling time will be used for generating headline results. According 
to EMA guidilines on GUIDELINE ON THE INVESTIGATION OF 
BIOEQUIVALENCE (2010, Doc. Ref.: [CPMP/EWP/QWP/1401/98 Rev. 1/ Corr **. 
2010]), following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using fixed effect ANOVA model with fixed 
effect terms for sequence, subject within sequence, period and treatment 
(formulation).
As a sensitivity analysis, mixed effect model with fixed effect terms for period, 
sequence and treatment (formulation), and subject as random effect term (i.e. ANOVA 
method using period, sequence and treatment as fixed effect, and subject as random 
effect) will be performed.
Point estimates and their associated 90% confidence intervals will be calculated for the 
differences: F1 - R, F2 - R, F3 -R, F4 - R for Part 1; FG1-R, FG2-R, FG3-R for Part 2; 
X1-R1, X2-R2 for Part 3A; and Y1-R3, Y2-R4 for Part 3B. The point estimates and their 
associated 90% confidence intervals will then be back-transformed to provide point 
estimates and 90% confidence intervals for the ratios, F1:R, F2:R, F3:R, F4:R; for Part 1; 
FG1:R, FG2:R, FG3:R for Part 2; X1:R1,X2:R2 for Part3A; and Y1:R3 and Y2:R4 for 
Part 3B.
 
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TITLE PAGE
Division: Worldwide Development
Information Type: Protocol Amendment
Title:
A Phase 1 study to demonstrate the relative bioavailability and 
bioequivalence of fixed dose combinations of ambrisentan and 
tadalafil in healthy subjects
Compound Number:
GSK3380154, GSK1325760 and GF196960
Development Phase:    I 
Effective Date:
08-FEB-2017
Protocol Amendment Number:  02
Author (s):
(Clinical Development Manager), 
(Medical Advisor), 
(Principal Statistician), 
(Sr. Director Clin. 
Pharmacology) , 
(Principle Data Manager), 
(Medical 
Director SERM), 
(SM Manager), 
(Supply Chain Study Lead)
Copyright 2017 the GlaxoSmithKline group of companies.  All rights reserved.  
Unauthorised copying or use of this information is prohibited.
 
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Revision Chronology:
GlaxoSmithKline
Document Number
Date
Version
2015N232335_00
2015-DEC-16
Original
2015N232335_01
2016-APR-14
Amendment No. 1
Clarification of hypotension withdrawal criteria in Section 5.4.3, change to GSK medical 
monitor and minor administrative changes.
2015N232335_02
2017-FEB-08
Amendment No. 2

Inclusion of bioequivalence assessment in Part 3 to replace the optional food 
effect. 

Inclusion of additional dosage strength in Part 3. 

Change in Medical Monitor(s).
 
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MEDICAL MONITOR/SPONSOR INFORMATION PAGE
Medical Monitor/SAE Contact Information: 
Role
Name
Day Time Phone Number and 
email address
After-hours Phone/Cell/
Pager Number
Fax 
Number
Primary 
Medical 
Monitor
Site to 
scan and 
send to 
email
Secondary 
Medical 
Monitor
Site to 
scan and 
send to 
email
SAE 
contact  
information 
Site to 
scan and 
send to 
email
Sponsor Legal Registered Address: 
GlaxoSmithKline Research & Development Limited
980 Great West Road
Brentford
Middlesex, TW8 9GS
UK
In some countries, the clinical trial sponsor may be the local GlaxoSmithKline Affiliate 
Company (or designee). If applicable, the details of the alternative Sponsor and contact 
person in the territory will be provided to the relevant regulatory authority as part of the 
clinical trial application.   
Regulatory Agency Identifying Number(s): EudraCT number 2015-004140-18
 
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INVESTIGATOR PROTOCOL AGREEMENT PAGE
For protocol number 201964
I confirm agreement to conduct the study in compliance with the protocol, as amended by 
this protocol amendment.
I acknowledge that I am responsible for overall study conduct. I agree to personally 
conduct or supervise the described study.
I agree to ensure that all associates, colleagues and employees assisting in the conduct of 
the study are informed about their obligations. Mechanisms are in place to ensure that site 
staff receives the appropriate information throughout the study. 
Investigator Name:
Investigator Address:
Investigator Phone Number:
Investigator Signature
Date
 
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TABLE OF CONTENTS
PAGE
1.
PROTOCOL SYNOPSIS FOR STUDY 201964........................................................9
2.
INTRODUCTION....................................................................................................13
2.1.
Study Rationale ..........................................................................................13
2.2.
Brief Background ........................................................................................14
3.
OBJECTIVE(S) AND ENDPOINT(S) ......................................................................16
4.
STUDY DESIGN ....................................................................................................17
4.1.
Overall Design ............................................................................................17
4.2.
Treatment and Duration..............................................................................18
4.3.
Type and Number of Subjects.....................................................................23
4.4.
Design Justification.....................................................................................23
4.5.
Dose Justification........................................................................................23
4.6.
Benefit: Risk Assessment ...........................................................................24
4.6.1.
Risk Assessment .........................................................................24
4.6.2.
Benefit Assessment.....................................................................26
4.6.3.
Overall Benefit: Risk Conclusion..................................................26
5.
SELECTION OF STUDY POPULATION AND WITHDRAWAL CRITERIA .............26
5.1.
Inclusion Criteria.........................................................................................27
5.2.
Exclusion Criteria........................................................................................28
5.3.
Screening/Baseline/Run-in Failures............................................................29
5.4.
Withdrawal/Stopping Criteria.......................................................................30
5.4.1.
Liver Chemistry Stopping Criteria ................................................30
5.4.1.1.
Study Treatment Restart or Rechallenge....................31
5.4.2.
QTc Stopping Criteria ..................................................................31
5.4.3.
Hypotension.................................................................................32
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria .........................32
5.5.
Subject and Study Completion....................................................................32
6.
STUDY TREATMENT ............................................................................................32
6.1.
Investigational Product and Other Study Treatment....................................32
6.1.1.
Retention Samples ......................................................................38
6.2.
Medical Devices..........................................................................................38
6.3.
Treatment Assignment................................................................................38
6.4.
Blinding.......................................................................................................40
6.5.
Packaging and Labeling..............................................................................40
6.6.
Preparation/Handling/Storage/Accountability..............................................40
6.7.
Compliance with Study Treatment Administration.......................................40
6.8.
Treatment of Study Treatment Overdose....................................................40
6.9.
Treatment after the End of the Study ..........................................................41
6.10.
Lifestyle and/or Dietary Restrictions............................................................41
6.10.1.
Meals and Dietary Restrictions ....................................................41
6.10.2.
Caffeine, Alcohol, and Tobacco ...................................................42
6.10.3.
Activity .........................................................................................42
6.11.
Concomitant Medications and Non-Drug Therapies....................................42
6.11.1.
Permitted Medications and Non-Drug Therapies..........................42
 
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6.11.2.
Prohibited Medications and Non-Drug Therapies.........................42
7.
STUDY ASSESSMENTS AND PROCEDURES .....................................................42
7.1.
Time and Events Table...............................................................................44
7.2.
Screening and Critical Baseline Assessments ............................................45
7.2.1.
Demographic/Medical History Assessments ................................45
7.3.
Pharmacokinetics .......................................................................................45
7.3.1.
Blood Sample Collection..............................................................45
7.3.2.
Sample Analysis ..........................................................................45
7.4.
Safety .........................................................................................................46
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)..........46
7.4.1.1.
Time period and Frequency for collecting AE 
and SAE information...................................................46
7.4.1.2.
Method of Detecting AEs and SAEs ...........................47
7.4.1.3.
Follow-up of AEs and SAEs........................................47
7.4.1.4.
Regulatory Reporting Requirements for SAEs............47
7.4.2.
Physical Exams ...........................................................................47
7.4.3.
Vital Signs....................................................................................48
7.4.4.
Electrocardiogram (ECG).............................................................48
7.4.5.
Clinical Safety Laboratory Assessments ......................................48
8.
DATA MANAGEMENT ...........................................................................................50
9.
STATISTICAL CONSIDERATIONS AND DATA ANALYSES .................................50
9.1.
Hypotheses.................................................................................................50
9.2.
Sample Size Considerations.......................................................................51
9.2.1.
Sample Size Assumptions ...........................................................51
9.2.2.
Sample Size Sensitivity................................................................52
9.2.3.
Sample Size Re-estimation or Adjustment...................................53
9.3.
Data Analysis Considerations .....................................................................53
9.3.1.
Analysis Populations....................................................................54
9.3.1.1.
Safety Population .......................................................54
9.3.1.2.
Pharmacokinetic Concentration Population ................54
9.3.1.3.
Pharmacokinetic Parameter Population......................54
9.3.2.
Interim Analysis ...........................................................................54
9.4.
Key Elements of Analysis Plan ...................................................................54
9.4.1.
Pharmacokinetic Analyses...........................................................54
9.4.2.
Safety Analyses...........................................................................55
10. STUDY GOVERNANCE CONSIDERATIONS........................................................56
10.1.
Posting of Information on Publicly Available Clinical Trial Registers............56
10.2.
Regulatory and Ethical Considerations, Including the Informed 
Consent Process ........................................................................................56
10.3.
Quality Control (Study Monitoring) ..............................................................57
10.4.
Quality Assurance.......................................................................................57
10.5.
Study and Site Closure ...............................................................................57
10.6.
Records Retention......................................................................................58
10.7.
Provision of Study Results to Investigators, Posting of Information 
on Publically Available Clinical Trials Registers and Publication .................58
11. REFERENCES.......................................................................................................60
 
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12. APPENDICES........................................................................................................62
12.1.
Appendix 1: Abbreviations and Trademarks................................................62
12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments ..............................................................................................63
12.3.
Appendix 3: Definition of and Procedures for Recording, Evaluating, 
Follow-Up and Reporting of Adverse Events...............................................65
12.3.1.
Definition of Adverse Events........................................................65
12.3.2.
Definition of Serious Adverse Events...........................................66
12.3.3.
Definition of Cardiovascular Events .............................................67
12.3.4.
Recording of AEs and SAEs ........................................................68
12.3.5.
Evaluating AEs and SAEs............................................................68
12.3.6.
Reporting of SAEs to GSK...........................................................70
12.4.
Appendix 4 High Fat Meal Content .............................................................71
12.5.
Appendix 5: Protocol Changes....................................................................72
12.5.1.
Protocol changes for Amendment 01 (14-Apr-2016) from 
Original Protocol (16-Dec-2015) ..................................................72
12.5.2.
Protocol changes for Amendment 2 () from Amendment 
1(14-Apr-2016) ............................................................................74
 
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1.
PROTOCOL SYNOPSIS FOR STUDY 201964
Rationale
This study is designed initially to compare the relative bioavailability of a number of 
fixed dose combinations (FDCs) of ambrisentan and tadalafil (Part 1 and 2) and 
consequently the bioequivalence of the FDC of different dose strength (Part 3A and 3B).
Dependent on formulation work, the study will allow up to 9 new FDCs to be compared 
with the reference of ambrisentan and tadalafil monotherapies taken concurrently.
Objective(s)/Endpoint(s)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following administration 
of candidate tablet formulations of the FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), and 
AUC(0-t) of ambrisentan and tadalafil in FDC 
and reference treatments
To establish bioequivalence of the candidate FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fed and fasted conditions.
AUC(0-t),AUC(0-inf) ,Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC (0-t), AUC(0-inf) ,Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 20 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 20 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC (0-t), AUC(0-inf) ,Cmax
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fed and fasted conditions.
Plasma PK parameters including; AUC(0-
t)/AUC(0-inf) ,tmax, t1/2 of ambrisentan and 
tadalafil in FDC and reference treatments
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fed and fasting
conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
 
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Overall Design
This is a single centre, Phase 1, single dose, randomised, open label crossover study in 
healthy volunteers with 3 study parts. Part 1 will include a 5 way cross-over. Part 2 and 
Part 3 (A&B) will each include a 4 way cross-over. All subjects will attend the unit for 
Screening within 31 days of their first dose. Eligible subjects will be randomised to order 
of treatment in that study parts in which they are included and the order in which these 
treatments are administered will be in accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in Parts 1 and 2. In Part 3 
the washout will be at least 10 days.  A Follow-Up Visit should be completed within 7-14 
days of completion of a subject’s last dose.  The utility of each of the 3 study parts is as 
follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part will include an evaluation of the bioavailability, safety and tolerability of 
3 granulation forms of a single FDC (ambrisentan 10mg + tadalafil 40 mg) identified 
from Part 1. These data will be compared to that for the reference of the 2 monotherapy 
components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All doses are 
taken in the fasted state.
Part 3 A
Part 3A of the study is set to establish bioequivalence between the candidate FDC from 
Part 2. This study part will have 4 dose sessions and will assess the bioequivalence, in 
both the fed and fasted state, of the FDC against the reference ambrisentan 10 mg + 
tadalafil 40mg monotherapies in the fed and fasted state.  The fed arms of this part will 
have a standard high fat breakfast. (EMEA, 2010).
Part 3 B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 40mg and 
ambrisentan 5 mg + tadalafil 20mg). This study part will have 4 dose sessions and will
assess the bioequivalence of the two FDC dose strengths against the reference
 
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ambrisentan 5 mg + tadalafil 40mg and ambrisentan 5 mg + tadalafil 20mg 
monotherapies in the fasted state.
Treatment Arms and Duration
The proposed treatment arms for each study part are described here.
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)
Part 3A (fed and fasted)
Part 3B 
(fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 1
(10mg/40mg)
ambrisentan and tadalafil 
FDC ) (10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 2
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg) fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 3
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
 
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Type and Number of Subjects
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable subjects 
complete each part of the study.  Approximately 33 healthy subjects will be randomized
to study Part 3A, such that at least 26 evaluable subjects complete part 3A of the study. 
An additional 33 healthy subejcts will be randomised to Part 3B, such that at least 26 
evaluable subjects complete part 3B of the study.
Analysis
No formal hypothesis will be tested for Part 1 and Part 2 of the study. An estimation 
approach will be used to estimate the bioavailability of the FDC formulations relative to 
the monotherapies taken concurrently.  
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 10 
mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 10 mg & 
tadalafil 40 mg taken concurrently (reference) in healthy subjects in both the fed and 
fasted states. The null hypothesis is that the true ratio of the geometric mean of the test 
treatment to the geometric mean of the reference treatment, (test)/(reference), for each 
primary PK endpoint, is either less than 0.80 or greater than 1.25.  The alternate 
hypothesis is that the true ratio of the test treatment geometric mean to the reference 
treatment geometric mean is at least 0.80 and no greater than 1.25.  Symbolically, this is 
expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e, treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test (TOST) 
procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be used to test 
this set of hypotheses. This is equivalent to requiring that a 90% interval for the true ratio 
of test to reference geometric means falls entirely within the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test the bioequivalence of a
FDC of ambrisentan 5 mg + tadalafil 40 mg (test) and of ambrisentan 5 mg + tadalafil 20 
mg (test) relative to reference monotherapies of  ambrisentan 5 mg & tadalafil 40 mg and 
of ambrisentan 5 mg & tadalafil 20 mg respectively taken concurrently (reference) in 
 
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healthy subjects in the fasted states. Therefore, the analysis approach for bioequivalence 
for Part 3A is  applicable to Part 3B.
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
2.
INTRODUCTION
Ambrisentan (E.U. trade name: Volibris), is an orally active endothelin receptor 
antagonist (ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first 
approved on 15 June 2007 in the US and on 21 April 2008 in the European Union (EU) 
and is currently approved in over 50 countries. In the EU, ambrisentan is indicated for 
treatment of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional 
Class (FC) II to III, including use in combination treatment (Volibris EMA SmPc, 2015;
Section 5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated 
with connective tissue disease.
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (EMEA, 2010;Schuirmann, 1987;Adcirca, 2015).
A recently completed study (Galiè, 2015a) has shown that patients with PAH who started 
initial combination therapy with ambrisentan and tadalafil had a significantly lower risk 
of clinical-failure events compared to those that started with ambrisentan or tadalafil 
monotherapy. Ambrisentan has recently received EU approval (20 November 2015) for 
use in combination treatment with tadalafil (Volibris EMA SmPc, 2015, Section 5.1).
2.1.
Study Rationale
This study is designed initially to compare the relative bioavailability of a number of 
fixed dose combinations (FDCs) of ambrisentan and tadalafil (Part 1 and 2) and 
consequently the bioequivalence of the FDC of different dose strengths (Part 3A and 3B).
Dependent on formulation work, the study will allow up to 9 new FDCs to be compared 
with the reference of ambrisentan and tadalafil monotherapies taken concurrently
The formulation(s) to be taken forward in Part 3A will be primarily chosen based on the 
test/reference ratio for both AUC and Cmax for both components. Ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, tablet size, cost, ease of 
manufacture and stability would be considered. 
Part 3 will include evaluation of bioequivalence for 3 dose strengths of the FDC; 
10/40mg, 5/40mg and 5/20mg of ambrisentan and tadalafil.  
 
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2.2.
Brief Background
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris EMA SmPC 2015, Section 5.1). Efficacy has been 
shown in idiopathic PAH (IPAH) and in PAH associated with connective tissue disease
(Volibris EMA SmPC, 2015). Ambrisentan is an oral, once daily, propanoic acid-based, 
ETA-selective Endothelin receptor antagonist (ERA) which targets the phospholipase-C-
dependent endothelin pathway and which is known to play an essential role in 
mammalian cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (EMEA, 2010;Schuirmann, 
1987;Adcirca, 2015). Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) 
inhibitor which targets the NO pathway. Through inhibition of PDE-5, tadalafil increases 
cytoplasmic cGMP concentrations in the smooth muscle cells and enhances NO-mediated 
vasodilatation of the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy participants was performed to 
detect any significant PK interactions between tadalafil and ambrisentan when co-
administered (Spence, 2009). From this study, it was concluded that there is no clinically 
significant PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when 
combined. Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
 
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administered.  There were no SAE’s in the study. Three participants withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one participantbecause of myalgia 
(severe), muscle fatigue and dizziness on tadalafil alone and one because of headache 
(severe) on the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild 
and is a listed event for ambrisentan. There were a total of 7 participants with mild 
tachycardia. There were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. 
There were 3 events on the first day of tadalafil 40 mg given 3 days after ambrisentan 10 
mg. There was one event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of 
ambrisentan 10 mg. Taken together these data suggest that in healthy subjects a mild 
tachycardia may result from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris EMA SmPC, 
2015, (EMEA, 2010;Schuirmann, 1987;Adcirca, 2015).
The AMBITION clinical study (Galiè, 2015a and GSK document number 
2014N193963_00), which evaluated the time to first clinical failure event, a composite 
endpoint, shows a robust clinical benefit (50% hazard reduction) for PAH patients 
initiated on a combination of ambrisentan and tadalafil when compared to PAH patients 
initiated on either medication as monotherapy.  The safety profile of the combination arm 
was consistent with the known safety data of the individual study drugs and no safety 
signals specific to combination treatment were identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 2008).
 
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3.
OBJECTIVE(S) AND ENDPOINT(S)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following administration 
of candidate tablet formulations of the FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), and 
AUC(0-t) of ambrisentan and tadalafil in FDC 
and reference treatments
To establish bioequivalence of the candidate FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fed and fasted conditions.
AUC(0-t), AUC(0-inf), Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC(0-t), AUC (0-inf), Cmax
To establish bioequivalence of the FDC 
(ambrisentan 5 mg + tadalafil 20 mg) relative to 
reference monotherapies tested (ambrisentan 5 mg 
& tadalafil 20 mg) taken concurrently in healthy 
human subjects under fasted conditions.
AUC (0-t), AUC (0-inf) ,Cmax
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting and fed conditions.
Plasma PK parameters including; AUC(0-
t)/AUC(0-inf), tmax, t1/2 of ambrisentan and 
tadalafil in FDC and reference treatments
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fed and fasting 
conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
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4.
STUDY DESIGN
4.1.
Overall Design
This is a single centre, Phase 1, single dose, randomised, open label, crossover study in 
healthy volunteers with 3 study parts; Part 1 will include a 5 way cross-over, and Part 2 
and 3 (A&B) will each include a 4 way cross-over. See Figure 1 for study schematic.
Figure 1
Study Schematic
Screening
≤ 31 days
Follow 
Up
Screening 
≤ 31 days from
Subjects 
Randomised to  
4 dose sessions 
(3 granulation 
formulations + 
reference)
Follow 
Up
Subjects 
randomised to 4 
dose sessions 
(FDC Fed & 
fasted + 
reference (fed 
&fasted))
Screening 
≤ 31 days
Follow 
Up
~4 Weeks
~4 Weeks
~4 Weeks
PART 1 (BA) n=20
PART 2 (BA) n=20
PART 3A (BE 10/40) n=33
Subjects 
Randomised to 
up to 5 dose 
sessions (4 FDC 
formulations 
and the 
reference)
Screening 
≤ 31 days
Subjects 
Randomised to 
4 dose sessions    
(2 FDC 
formulations 
(fasted) + 2 
reference fasted)
Follow 
Up
PART 3B (BE 5/40 & 5/20)
n=33
~4 Weeks
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in study Part 1 and 2. In
study Part 3 there will be a minimum of 10 days between each dose.  A Follow-Up Visit 
should be completed within 7-14 days of completion of a subject’s last dose. 
The utility of each of the 3 study parts is as follows:
 
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Part 1
This study part, will have 5 dose sessions and will be used  to characterise the relative 
bioavailability, safety and tolerability of   four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
Part 2
This study part will have 4 dose sessions and will evaluate the bioavailability, safety and 
tolerability of  3 different granulation sizes for a single FDC (ambrisentan 10 mg + 
tadalafil 40 mg) compared to the reference of the 2 monotherapy components taken 
concurrently (ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted 
state.
Part 3A
Part 3A of the study is set to establish bioequivalence between the candidate FDC from 
Part 2. This study part will have 4 dose sessions and will assess the bioequivalence, in 
both the fed and fasted state, of the FDC against the reference  ambrisentan 10 mg + 
tadalafil 40mg monotherapies in the fed and fasted state. The fed arms of this part will 
have a standard high fat breakfast (EMEA, 2010).
Part 3B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 40mg and 
ambrisentan 5 mg + tadalafil 20mg) . This study part will have 4 dose sessions and will
assess the bioequivalence of the two FDC dose strengths against the reference 
ambrisentan 5 mg + tadalafil 40mg monotherapies and ambrisentan 5 mg + tadalafil 
20mg monotherapies in the fasted state.
4.2.
Treatment and Duration
The treatments for each study part of the study are listed in Table 1. All treatments are 
single dose. Subjects will be randomised to the order of treatments in the parts of the 
study they are included in.
The study has 3 parts and ongoing analysis of pharmacokinetic data will be used to 
determine the formulations produced and tested in subsequent parts.
Part 1
Part 1 of the study will be utilised to evaluate 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
 
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following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2
and 3. 
Following completion of Part 1 there will be a pause prior to Part 2, so that different 
granulation forms of a single FDC can be produced and data included and approved in 
any required update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for 3 granulation size forms of a single FDC 
formulation selected from Part 1. These will be based on a single successful formulation 
identified in Part 1 reformulated with the final active pharmaceutical ingredient (API).
Success will be defined with the same criteria as those in Part 1.
Part 3 (A and B)
The BE part of the study will investigate the bioequivalence of the FDC ambrisentan 10 
mg + tadalafil 40 mg in part 3A and the bioequivalence of the FDC ambrisentan 5 mg + 
tadalafil 40 mg and ambrisentan 5 mg + tadalafil 20 mg in Part 3B.  
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted) 
Part 3A (fed and fasted)
Part 3B (fasted)
ambrisentan and
tadalafil FDC1 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 1
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and tadalafil 
FDCgranulation 2
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg) 
fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 3
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg), fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg), fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
The duration for a subject on this study is described in Figure 2. This figure assumes all 5 
dose sessions are used for Part 1. However, this is an approximation as minimum of 7 
days is expected between doses in Parts 1 and 2 and a minimum of 10 days in Part 3.
 
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Figure 2
Study Duration
Part 1 can last up to 73 days.
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
Dose 
Period 5 
start 
7 
days
7 
days
7 
days
7 
days
7 to 
14 
days
Follow-up
 
 
 
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Part 2 will last approximately 9 weeks
Part 3A and B will last approximately 11 weeks
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
10 
days
10  
days
10 
days
7 to 14  
days
Follow-up
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
7 days
7 days
7 days
7 to 14  
days
Follow-up
 
 
 
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A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
4.3.
Type and Number of Subjects
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable subjects 
complete each part of the study.  Approximately 33 healthy subjects will be randomized
to study Part 3A, such that at least 26 evaluable subjects complete part 3A of the study. 
An additional 33 healthy subjects will be randomised to Part 3B, such that at least 26 
evaluable subjects complete part 3B of the study.
The estimation of the number of subjects for each study part is detailed further in Section 
9.2 Sample Size Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
4.4.
Design Justification
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, nor is there any need to blind study treatment. The inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study contains three parts: Parts 1 and 2 will provide PK data for different candidate 
FDC formulations and will enable the assessment of bioequivalence of the FDC versus 
the combination of the individual reference products in Part 3. The interdependency of 
the 3 study parts is described in detail in Section 4.2. 
4.5.
Dose Justification
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of 40mg for tadalafil and 10mg for ambrisentan, are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
 
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be tolerated in healthy subjects. Within Part 3 of the study all dosage forms of the FDC 
that are proposed for marketing authorisation will be tested for bioequivalence versus the 
appropriate doses of reference products.  These doses are required in clinical practice to 
allow safe up- and down-titration of the FDC in patients with PAH.  
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimumof 10 days, between doses, 
is also sufficient for clearance of the previous dose during the assessmet of 
bioequivalence.
4.6.
Benefit: Risk Assessment
There is no direct benefit for healthy subjects participating in this study.  The risks to the 
healthy subjects based upon previous experience indicate no expectation of SAE and mild 
to moderate AE.  The benefit/risk remains satisfactory for this healthy subject study.
Summaries of findings from clinical studies conducted with both the Investigational 
Product taken as monotherapies and in combination can be found in: 

Ambrisentan IB (Volibris IB, 2015)

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015) 

Adcirca (Tadalafil) EMA SmPC (EMEA, 2010;Schuirmann, 1987;Adcirca, 2015)
As described in Section 2.2, mild tachycardia was the most common (7/26, 27%) adverse 
event seen in healthy subjects following combination dosing with 10mg ambrisentan and 
40mg tadalafil (Spence, 2009). There were three subjects (3/26, 11.5%) withdrawn due to 
AEs (e.g., anaemia [mild]; myalgia [severe], muscle fatigue, and dizziness; and headache 
[severe]).    No SAEs or additional risks were observed with combination use in healthy 
subjects.
The following section outlines the risk assessment and mitigation strategy for this 
protocol:  
4.6.1.
Risk Assessment
Risks of clinical significance, identified in subjects with PAH and the mitigation strategy 
identified for this study in healthy subjects are captured in Table 2. 
 
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Table 2
Risks of Clinical Significance and Mitigation Strategy
Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
Investigational Product: GSK1325760 (ambrisentan-Volibris)

Teratogenicity

Anaemia (decreased 
haemoglobin, decreased 
haematocrit)

Hypersensitivity reactions 
(e.g. angioedema, rash, 
pruritus)

Headache (including sinus 
headache, migraine)

Dizziness

Cardiac failure

Palpitation

Hypotension

Flushing

Epistaxis

Dyspnoea

Upper respiratory  
congestion, sinusitis, 
nasopharyngitis, rhinitis

Abdominal pain

Constipation

Nausea, vomiting, 
diarrhoea

Hepatic transaminases 
increased

Peripheral oedema, fluid 
retention

Chest pain/discomfort

Asthenia and fatigue
Ambrisentan IB (Volibris IB, 
2015)
Volibris (Ambrisentan) EMA 
SmPC (Volibris EMA SmPC, 
2015): 
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision. 

Exclusion of women of 
childbearing potential 
(teratogenicity)

Laboratory assessments 
per protocol

Physical assessment per 
protocol

Routine vital signs per 
protocol

Subjects remain in the 
clinical unit, under medical 
supervision, for all doses 
and until completion of 
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GF196960 (tadalafil - Adcirca)

Hypersensitivity reactions

Headache

Syncope,

Migraine

Blurred vision

Palpitations

Nasopharyngitis (including 
nasal congestion, sinus 
congestion and rhinitis)

Epistaxis
Adcirca (Tadalafil) EMA SmPC 
(EMEA, 2010;Schuirmann, 
1987;Adcirca, 2015)
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision.

Routine vital signs per 
protocol

Physical assessment per 
protocol

Subjects remain in the 
clinical unit for all doses, 
under medical supervision 
and until completion of 
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy

Nausea,

Dyspepsia (including 
abdominal pain/discomfort)

Vomiting,

Gastroesophageal reflux

Rash

Myalgia,

Back pain

Pain in extremity (including 
limb discomfort)

Increased uterine bleeding

Facial oedema,

Chest pain
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GSK3380154 (ambrisentan-tadalafil-FDC)
The risks for the combination 
are the same as the 2 
monotherapies; no extra risks 
have been identified for the 
combination.
As for GSK1325760 and 
GF196960 above
4.6.2.
Benefit Assessment
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation and to 
demonstrate bioequivalence versus reference products for a fixed dose combination of 
ambrisentan plus tadalafil to use as first line therapy in PAH patients.
4.6.3.
Overall Benefit: Risk Conclusion
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 2. In 
addition review of safety information from Part 1 of the study is consistent with this 
position. So, the benefit:risk is appropriate for this study and to enable development of 
this investigational product. The overall benefit: risk therefore remains positive.
5.
SELECTION OF STUDY POPULATION AND 
WITHDRAWAL CRITERIA
Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in:

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015)
 
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
Adcirca (Tadalafil) EMA SmPC (EMEA, 2010;Schuirmann, 1987;Adcirca, 2015)
Deviations from inclusion and exclusion criteria are not allowed because they can 
potentially jeopardize the scientific integrity of the study, regulatory acceptability or 
subject safety. Therefore, adherence to the criteria as specified in the protocol is essential.
5.1.
Inclusion Criteria
A subject will be eligible for inclusion in this study only if all of the following criteria 
apply:
AGE
1. Between 18 and 60 years of age inclusive, at the time of signing the informed 
consent.
TYPE OF SUBJECT AND DIAGNOSIS INCLUDING DISEASE SEVERITY
2. Healthy as determined by a responsible and experienced physician, based on a 
medical evaluation including medical history, physical examination, laboratory tests, 
vital signs and cardiac monitoring (ECG and 24 hour Holter).  A subject with a 
clinical abnormality or laboratory parameter(s) which is/are not specifically listed in 
the inclusion or exclusion criteria, outside the reference range for the population 
being studied may be included only if the Investigator, in consultation with the GSK 
Medical Monitor if required, judges and documents that the finding is unlikely to 
introduce additional risk factors and will not interfere with the study procedures. 
WEIGHT
3. Body weight ≥ 50 kg (110 lbs) for men and ≥ 45 kg (99lbs) for women and body 
mass index (BMI) within the range 18– 30 kg/m2 (inclusive)
SEX
4. Male or Female
Females must be the following:
Non-reproductive potential defined as:

Pre-menopausal females with one of the following:
Documented tubal ligation
Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion 
Hysterectomy
Documented Bilateral Oophorectomy
 
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
Documented Postmenopausal defined as 12 months of spontaneous amenorrhea
INFORMED CONSENT
5. Capable of giving signed informed consent as described in Section 10 which includes 
compliance with the requirements and restrictions listed in the consent form and in 
this protocol.
5.2.
Exclusion Criteria
A subject will not be eligible for inclusion in this study if any of the following criteria 
apply:
CONCURRENT CONDITIONS/MEDICAL HISTORY (INCLUDES LIVER 
FUNCTION AND QTC INTERVAL)
1.
A blood pressure <100/55 mm Hg.
2.
Haemoglobin below normal range:

Hb < 133 g/L for males 

Hb< 114 g/L for females
3.
ALT and bilirubin >1.5xULN (isolated bilirubin >1.5xULN is acceptable if bilirubin 
is fractionated and direct bilirubin <35%). 
4.
Current or chronic history of liver disease, or known hepatic or biliary abnormalities 
(with the exception of Gilbert's syndrome or asymptomatic gallstones)
5.
QTc> 450 msec
NOTES:

The QTc is the QT interval corrected for heart rate according to Bazett’s 
formula (QTcB), Fridericia’s formula (QTcF), and/or another method, machine-
read or manually over-read.

The specific formula that will be used to determine eligibility and 
discontinuation for an individual subject should be determined prior to initiation 
of the study. In other words, several different formulae cannot be used to 
calculate the QTc for an individual subject and then the lowest QTc value used 
to include or discontinue the subject from the trial.

For purposes of data analysis, QTcB, QTcF, another QT correction formula, or a 
composite of available values of QTc will be used as specified in the Reporting 
and Analysis Plan (RAP).
CONCOMITANT MEDICATIONS
6.
Use of prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements (including St John’s Wort) within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose 
 
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of study medication, unless in the opinion of the Investigator and GSK Medical 
Monitor the medication will not interfere with the study procedures or compromise 
subject safety. 
RELEVANT HABITS
7.
History of regular alcohol consumption within 6 months of the study defined as: 

An average weekly intake of >21 units for males or >14 units for females.  One 
unit is equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 
ml) of wine or 1 (25 ml) measure of spirits.
8.
Smoking more than 5 cigarettes per week and subjects must be able to abstain from 
smoking for a 24 hour period prior to dose and any time whilst in the clinical unit.
CONTRAINDICATIONS
9.
History of sensitivity to any of the study medications, or components thereof or a 
history of drug or other allergy that, in the opinion of the Investigator or Medical 
Monitor, contraindicates their participation.
DIAGNOSTIC ASSESSMENTS AND OTHER CRITERIA
10. Presence of hepatitis B surface antigen (HBsAg), positive hepatitis C antibody test 
result at Screening or within 3 months prior to first dose of study treatment. . 
11. A positive test for HIV antibody.
12. A positive pre-study drug/alcohol screen.  
13. Where participation in the study would result in donation of blood or blood products 
in excess of 500 mL within previous 3 months
14. The subject has participated in a clinical trial and has received an investigational 
product within the following time period prior to the first dosing day in the current 
study: 3 months, 5 half-lives or twice the duration of the biological effect of the 
investigational product (whichever is longer).
15. Exposure to more than four new chemical entities within 12 months prior to the first 
dosing day.
5.3.
Screening/Baseline/Run-in Failures
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT)
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
 
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set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.4).
5.4.
Withdrawal/Stopping Criteria
The following actions must be taken in relation to a subject who fails to attend the clinic 
for a required study visit:

The site must attempt to contact the subject and re-schedule the missed visit as 
soon as possible. 

The site must counsel the subject on the importance of maintaining the assigned 
visit schedule and ascertain whether or not the subject wishes to and/or should 
continue in the study. 

In cases where the subject is deemed ‘lost to follow up’, the investigator or 
designee must make every effort to regain contact with the subject (where 
possible, 3 telephone calls and if necessary a certified letter to the subject’s last 
known mailing address or local equivalent methods). These contact attempts 
should be documented in the subject’s medical record. 

Should the subject continue to be unreachable, only then will he/she be considered 
to have withdrawn from the study with a primary reason of “Lost to Follow-Up”. 
A subject may withdraw from study treatment at any time at his/her own request, or may 
be withdrawn at any time at the discretion of the investigator for safety, behavioural or 
administrative reasons. If a subject withdraws from the study, he/she may request 
destruction of any samples taken, and the investigator must document this in the site 
study records. 
5.4.1.
Liver Chemistry Stopping Criteria
Liver chemistry stopping and increased monitoring criteria have been designed to 
assure subject safety and evaluate liver event etiology (in alignment with the FDA 
premarketing clinical liver safety guidance). 
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf
 
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Study treatment will be discontinued for a subject if liver chemistry stopping criteria are 
met:
Phase I Liver Chemistry Stopping Criteria – Liver Stopping Event Algorithm
Continue Study Treatment 
Discontinue Study Treatment 
*INR value not applicable to subjects on anticoagulants 
ALT ≥3xULN 
No
Yes
Must refer to Liver Safety Required Actions and Follow up Assessments  section in the Appendix
Report as an SAE if possible Hy’s Law case: ALT≥3xULN and  Bilirubin≥2xULN (>35% direct)  or 
INR>1.5, if measured*
Liver Safety Required Actions and Follow-Up Assessments Section can be found in 
Appendix 2: Liver Safety Required Actions and Follow-Up Assessments
5.4.1.1.
Study Treatment Restart or Rechallenge
Study treatment restart or re-challenge after liver chemistry stopping criteria are met by 
any subject participating in this study is not allowed.
5.4.2.
QTc Stopping Criteria

The same QT correction formula must be used for each individual subject to 
determine eligibility for and discontinuation from the study.  This formula may 
not be changed or substituted once the subject has been enrolled.

For example, if a subject is eligible for the protocol based on QTcF, then QTcF 
must be used for discontinuation of this individual subject as well.

Once the QT correction formula has been chosen for a subject’s eligibility, the 
same formula must continue to be used for that subject for all QTc data being 
collected for data analysis.  Safety ECGs and other non-protocol specified 
ECGs are an exception.  

The QTc should be based on averaged QTc values of triplicate electrocardiograms 
obtained over a brief (e.g., 5-10 minute) recording period. 
A subject that meets either bulleted criterion below will be withdrawn from the study.  

QTc> 500 msec,
 
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
Change from baseline: QTc >60 msec 
5.4.3.
Hypotension
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 mmHG and diastolic <50 mm HG confirmed by 
triplicate reading taken up to 5 minutes apart and is judged clinically significant 
and symptomatic by the investigator.
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria
For an individual study participant, stopping criteria include, but are not limited to:
Adverse events, or significant changes in any of the safety assessments, that put the 
safety of the individual at risk (e.g., ECG, vital signs, laboratory tests, etc), as judged by 
the Principal Investigator in consultation with the Medical Monitor if necessary.
5.5.
Subject and Study Completion
A completed subject is one who has completed all dosing periods of the study including 
the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
6.
STUDY TREATMENT
6.1.
Investigational Product and Other Study Treatment
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Parts 1, 2 and 3 of the study are shown in Table 3, Table 
4 and Table 5. 
 
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Table 3
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 90 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.45
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Address of 
reference 
manufacturer
NA
NA
NA
NA
Glaxo Operations UK 
Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 4
Study Treatment for Part 2
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-tadalafil-
FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 granulation 1
(10mg/40mg)
FDC-G2 granulation 2
(10mg/40mg)
FDC-G3 granulation 3 
(10mg/40mg)
Each tablet contains 10 mg 
of ambrisentan, 
approximately 90 mg of 
lactose (as monohydrate), 
approximately 0.25 mg of 
lecithin (soya) (E322) and 
approximately 0.45 mg of 
Allura red AC Aluminium 
Lake 
Each film-coated tablet 
contains 20 mg 
tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage level(s):
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 40 
mg tadalafil and 10 mg 
of ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 10 mg 
of ambrisentan.
Each tablet contains 20 
mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations UK Ltd 
(trading as GlaxoWellcome 
Operations), Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la Industria 
30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 5
Study Treatment for Part 3
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-tadalafil-
FDC)
GSK1325760 
(ambrisentan-Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 or 2 or3
(10mg/40mg)
FDC  (5mg/40mg)
FDC  (5mg/20mg)
Each tablet contains 
10 mg of ambrisentan, 
approximately 90 mg 
of lactose (as 
monohydrate), 
approximately 0.25 
mg of lecithin (soya) 
(E322) and 
approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each tablet 
contains 5 mg of 
ambrisentan, 
approximately 95
mg of lactose (as 
monohydrate), 
approximately 0.25 
mg of lecithin 
(soya) (E322) and 
approximately 0.11 
mg of Allura Red
AC Aluminium 
Lake
Each film-coated 
tablet contains 20 
mg tadalafil, 245
mg lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 5 mg 
of ambrisentan.
Each tablet contains 
20 mg tadalafil and 5
mg of ambrisentan.
Each tablet contains 
10 mg of ambrisentan.
Each tablet 
contains 5 mg of 
ambrisentan.
Each tablet 
contains 20 mg 
tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-tadalafil-
FDC)
GSK1325760 
(ambrisentan-Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations UK 
Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Glaxo Operations 
UK Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 Alcobendas
Madrid
Spain
 
 
 
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6.1.1.
Retention Samples
GSK will send five times (5x) the drug product necessary for pivotal bioequivalency 
evaluation in Part 3 of the study for analytical testing for all tablet strengths used in the 
study. The retain samples are in addition to the supply of drug product sufficient to 
complete the study. Retention samples will be sent to the clinical CRO in the same bulk 
container. To ensure that reserve samples are representative of the same batches provided 
for the clinical study the clinical site will randomly select the supplies for the study from 
the supply received. The clinical site must retain enough reserve samples to permit FDA 
to perform five times all of the release tests required in the application.
6.2.
Medical Devices
No GSK manufactured devices (or devices manufactured for GSK by a third party) are 
provided for use in this study.
6.3.
Treatment Assignment
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software.
The treatment key for Parts 1, 2 and 3 are described in Table 6.
Table 6
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 2
FG1
ambrisentan and tadalafil FDC-G1 (10mg/40mg) granulation size 1
FG2
ambrisentan and tadalafil FDC-G2 (10mg/40mg) granulation size 2
FG3
ambrisentan and tadalafil FDC-G3 (10mg/40mg) granulation size 3
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3A
X1
ambrisentan and tadalafil FDC-G1 or 2 or 3 (10mg/40mg) fed
X2
ambrisentan and tadalafil FDC G1 or 2 or 3 (10mg/40mg) fasted
 
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Treatment
Description
R1
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fed
R2
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fasted
Part 3B
Y1
ambrisentan and tadalafil FDC (5mg/40mg), fasted
Y2
ambrisentan and tadalafil FDC (5mg/20mg), fasted
R3
ambrisentan and tadalafil monotherapies taken concurrently (5mg/40mg), 
fasted
R4
ambrisentan and tadalafil monotherapies taken concurrently (5mg/20mg), 
fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 7. Additional treatment sequences may
be created based using the Latin Squares for Williams Designs if needed.
Table 7
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1/ F2/ R/ F3/ F4
F2/ F3/ F1/ F4/ R
F3/ F4/ F2/ R/ F1
F4/ R/ F3/ F1/ F2
R/ F1/ F4/ F2/ F3
F4/ F3/ R/ F2/ F1
R/ F4/ F1/ F3/ F2
F1/ R/ F2/ F4/ F3
F2/ F1/ F3/ R/ F4
F3/ F2/ F4/ F1/ R
1:1:1:1:1:1:1:1:1:1
Part 2
4
FG1/ FG2/ R/ FG3
FG2/ FG3/ FG1/ R
FG3/ R/ FG2/ FG1
R/ FG1/ FG3/ FG2
1:1:1:1
Part 3A
4
X1/ R1/ R2/ X2
R1/ X2/ X1/ R2
X2/ R2/ R1/ X1
R2/ X1/ X2/ R1
1:1:1:1
Part 3B
4
Y1/ R3/ R4/ Y2
R3/ Y2/ Y1/ R4
Y2/ R4/ R3/ Y1
R4/ Y1/ Y2/ R3
1:1:1:1
 
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6.4.
Blinding
This will be an open-label study. 
6.5.
Packaging and Labeling
The contents of the label will be in accordance with all applicable regulatory 
requirements.
6.6.
Preparation/Handling/Storage/Accountability
This will be detailed in a Study Specific Technical Agreement/Memo (TTS) which will 
be accompanied by a Quality Agreement.

Only subjects enrolled in the study may receive study treatment and only 
authorized site staff may supply or administer study treatment.  All study 
treatments must be stored in a secure environmentally controlled and monitored 
(manual or automated) area in accordance with the labelled storage conditions 
with access limited to the investigator and authorized site staff. 

The investigator, institution, or the head of the medical institution (where 
applicable) is responsible for study treatment accountability, reconciliation, and 
record maintenance (i.e. receipt, reconciliation and final disposition records).  

Further guidance and information for final disposition of unused study treatment 
are provided in the SRM. 

Under normal conditions of handling and administration, study treatment is not 
expected to pose significant safety risks to site staff.  

A Material Safety Data Sheet (MSDS)/equivalent document describing 
occupational hazards and recommended handling precautions either will be 
provided to the investigator, where this is required by local laws, or is available 
upon request from GSK.
6.7.
Compliance with Study Treatment Administration
When subjects are dosed at the site, they will receive study treatment directly from the 
investigator or designee, under medical supervision.  The date and time of each dose 
administered in the clinic will be recorded in the source documents.  The dose of study 
treatment and study subject identification will be confirmed at the time of dosing by a 
member of the study site staff other than the person administering the study treatment.  
Study site personnel will examine each subject’s mouth to ensure that the study treatment 
was ingested.
6.8.
Treatment of Study Treatment Overdose
For this study, any dose of GSK3380154 (ambrisentan/tadalafil-FDC), GSK1325760 
(ambrisentan) and GF196960 (tadalafil) greater than the protocol defined dose and within 
a 24 hour time period will be considered an overdose.
 
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GSK does not recommend specific treatment for an overdose.
Advice for the investigator is included in the product label for GSK1325760 
(ambrisentan) and GF196960 (tadalafil).
In the event of an overdose the investigator should:
1.
Contact the Medical Monitor immediately
2.
Closely monitor the subject for adverse events (AEs)/serious adverse events (SAEs) 
and laboratory abnormalities until compound number/name can no longer be 
detected systemically (at least 3 days for compound number/name)
3.
Obtain a plasma sample for pharmacokinetic (PK) analysis if requested by the 
Medical Monitor (determined on a case-by-case basis)
4.
Document the quantity of the excess dose as well as the duration of the overdosing in 
the CRF.
Decisions regarding dose interruptions or modifications will be made by the investigator 
in consultation with the Medical Monitor based on the clinical evaluation of the subject.
6.9.
Treatment after the End of the Study
Subjects will not receive any additional treatment from GSK after completion of the 
study because only healthy subjects are eligible for study participation.
6.10.
Lifestyle and/or Dietary Restrictions
6.10.1.
Meals and Dietary Restrictions

Refrain from consumption of red wine, Seville oranges, grapefruit or grapefruit 
juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or fruit juices from 
7 days prior to the first dose of study medication until after the final dose.

Dependent on the utility of the dose session subjects will be asked to fast or 
consume the FDA full fat breakfast prior to dosing (EMEA, 2010)

Fasting subjects will be required to fast from midnight before each full in-house 
dosing day (i.e. Day 1) with the exception of water, which will be allowed freely 
except for 1 hour either side of dosing. Subjects will be required to fast up to 4h 
hour post dose on Day 1 of each dose

For treatments given under fed conditions, a standard high-fat breakfast will be 
provided before dosing. See Section 12.4 for details of this meal. This meal 
should be eaten in its entirety within 30 minutes.  The amount consumed will be 
recorded within the source documents and the CRF by the site staff.  Subjects 
will be dosed within 5 minutes of completing the breakfast.  

Subjects should take each dose of investigational product with 240 ml (8 fl oz) of 
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6.10.2.
Caffeine, Alcohol, and Tobacco

During each dosing session, subjects will abstain from ingesting caffeine- or 
xanthine-containing products (e.g., coffee, tea, cola drinks and chocolate) for 24 
hours prior to the start of dosing until collection of the final pharmacokinetic sample 
during each session. 

During each dosing session, subjects will abstain from alcohol for 24 hours prior to 
the start of dosing until collection of the final pharmacokinetic sample during each 
session. 

Smoking is not allowed for 24 hours prior to dosing and whilst subjects are in the 
clinic.
6.10.3.
Activity
Subjects will abstain from strenuous exercise from Screening until Follow-Up.  Subjects 
may participate in light recreational activities during studies (e.g., watch television, read).
6.11.
Concomitant Medications and Non-Drug Therapies
6.11.1.
Permitted Medications and Non-Drug Therapies
Paracetamol at doses of 2 grams/day is permitted for use any time during the study. 
Other concomitant medication may be considered on a case by case basis by the 
Investigator in consultation with the Medical Monitor if required.
6.11.2.
Prohibited Medications and Non-Drug Therapies
Subjects must abstain from taking prescription or non-prescription drugs (including 
vitamins and dietary or herbal supplements), within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose of 
study medication until completion of the Follow-Up visit, unless in the opinion of the 
Investigator and sponsor the medication will not interfere with the study.
7.
STUDY ASSESSMENTS AND PROCEDURES
Protocol waivers or exemptions are not allowed with the exception of immediate safety 
concerns. Therefore, adherence to the study design requirements, including those 
specified in the Time and Events Table, are essential and required for study conduct.
This section lists the procedures and parameters of each planned study assessment.  The 
exact timing of each assessment is listed in the Time and Events Table Section 7.1
The following points must be noted: 

If assessments are scheduled for the same nominal time, THEN the assessments 
should occur in the following order:
1. 12-lead ECG
2. Vital signs
 
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3. Blood draws.  
Note: The timing of the assessments should allow the blood draw to occur at the 
exact nominal time.

The timing and number of planned study assessments, safety and 
pharmacokinetic assessments may be altered during the course of the study 
based on newly available data (e.g., to obtain data closer to the time of peak 
plasma concentrations) to ensure appropriate monitoring. 

The change in timing or addition of time points for any planned study 
assessments must be documented in a Note to File which is approved by the 
relevant GSK study team member and then archived in the study sponsor and 
site study files, but this will not constitute a protocol amendment. 

The IRB/IEC will be informed of any safety issues that require alteration of the 
safety monitoring scheme or amendment of the Informed Consent Form. 
No more than 500 mL of blood will be collected over the duration of the study, including 
any extra assessments that may be required.
 
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7.1.
Time and Events Table
Procedure
Screen
Part 1, 2 and 3A and 3B. Each dose in each Part repeats this schedule.
FU
Notes
Day
≤-31
-1
1
2
3
4
≥7-14
Time (hrs)
Pre-
dose
0
0.5
1
1.5
2
2.5
4
8
12
18
24
36
48
72
Outpatient visit
x
x
x
Admission to unit
x
Informed consent
x
Inclusion and exclusion criteria
x
x
Demography
x
Full physical exam including 
height and weight
x
Brief Physical
x
Medical history (includes 
substance usage)
x
HIV, Hep B and Hep C screen] 
x
Laboratory assessments (include 
liver chemistries) 
x
x
x
x
Only Screening labs need to be taken in 
fasted state.
Serum  hCG Pregnancy test
x
x
Female subjects only
Urine  hCG Pregnancy test
x
Female subjects only
Breathalyser and Smokerlyzer
x
x
DOA testing
x
x
12-lead ECG
x
x
x
x
x
x
x
x
x
Triplicate at screen and baseline, single 
measure at other times, unless out of 
range then triplicates should be 
performed
Vital signs
x
x
x
x
x
x
x
x
x
x
x
x
24hr Holter
x
Randomisation
x
Randomised prior to first dose only
Study Treatment
x
AE/SAE review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
SAEs from Screen. AEs from first dose
Concomitant medication review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
PK Sample
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Discharge from Unit
x
For logistical reasons subjects may 
remain in-unit for the 72 hr assessments 
if they prefer.
 
 
 
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7.2.
Screening and Critical Baseline Assessments
All subjects must give written consent to participate in this trial.  Consent for screening 
evaluations may be obtained using the ICF for the Hammersmith Medicines Research 
(HMR) healthy subject’s panel, which has been approved by the HRA’s Phase 1 Advert 
Review group.  The study-specific information and consent form will be signed by the 
subject either before any screening evaluation or after the investigator confirms the 
eligibility of the subject for the study and before the subject is randomised to receive the 
first administration of IMP.  Before giving consent, subjects must read the information 
sheet about the study.  They must also read the consent form.  They will then discuss the 
study with the investigator or his deputy and be given the opportunity to ask questions.  
The study-specific information sheet and the consent form must be approved by the REC.
7.2.1.
Demographic/Medical History Assessments
Prior to enrolling in the study and having any study procedures completed, subjects must 
sign the informed consent.
During the Screening visit, each subject will undergo the assessments to determine 
eligibility for enrolment as detailed in Section 5. 
The following demographic parameters will be captured: date of birth, gender, race and 
ethnicity.
Medical/medication/alcohol history will be assessed as related to the eligibility criteria 
listed in Section 5.
7.3.
Pharmacokinetics
7.3.1.
Blood Sample Collection
Blood samples for pharmacokinetic (PK) analysis of ambrisentan and tadalafil will be 
collected at the time points indicated in Section 7.1, Time and Events Table.  The actual 
date and time of each blood sample collection will be recorded.  The timing of PK 
samples may be altered and/or PK samples may be obtained at additional time points to 
ensure thorough PK monitoring.
For analysis of ambrisentan 2.7 mL of blood will be collected into sodium citrate tubes 
and for analysis of tadalafil 2.0 mL of blood will be collected into K2-EDTA tubes. 
Processing, storage and shipping procedures are provided in the Study Reference Manual 
(SRM).
7.3.2.
Sample Analysis
Plasma analysis will be performed under the control of Platform Technology and Science 
In vitro/In vivo Translation (PTS IVIVT) and Third Party Resource, GlaxoSmithKline, 
the details of which will be included in the Study Reference Manual (SRM).
Concentrations of ambrisentan and tadalafil will be determined in plasma samples using 
 
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the currently approved bioanalytical methodology. Raw data will be archived at the
bioanalytical site (detailed in the SRM).
Once the plasma has been analyzed for ambrisentan and tadalafil any remaining 
plasma may be analyzed for other compound-related metabolites and the results reported 
under a separate PTS-IVIVT, GlaxoSmithKline protocol.
7.4.
Safety
Planned time points for all safety assessments are listed in the Time and Events Table 
(Section 7.1).  Additional time points for safety tests such as vital signs, physical 
examinations and laboratory safety assessments may be added during the course of the 
study based on newly available data to ensure appropriate safety monitoring.
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)
The definitions of an AE or SAE can be found in Appendix 3: Definition of and 
Procedures for Recording, Evaluating, Follow-Up and Reporting of Adverse Events.
The investigator and their designees are responsible for detecting, documenting and 
reporting events that meet the definition of an AE or SAE. 
7.4.1.1.
Time period and Frequency for collecting AE and SAE information

Any SAEs assessed as related to study participation (e.g., protocol-mandated 
procedures, invasive tests, or change in existing therapy) or related to a GSK 
product will be recorded from the time a subject consents to participate in the 
study up to and including any follow up contact. 

AEs will be collected from the start of Study Treatment until the follow up contact 
(see Section 7.4.1.3), at the timepoints specified in the Time and Events Table 
(Section 7.1).

Medical occurrences that begin prior to the start of study treatment but after 
obtaining informed consent may be recorded on the Medical History/Current 
Medical Conditions section of the CRF.

All SAEs will be recorded and reported to GSK within 24 hours, as indicated in 
Section 12.3.

Investigators are not obligated to actively seek AEs or SAEs in former study 
subjects. However, if the investigator learns of any SAE, including a death, at any 
time after a subject has been discharged from the study, and he/she considers the 
event reasonably related to the study treatment or study participation, the 
investigator must promptly notify GSK.
NOTE: The method of recording, evaluating and assessing causality of AEs and SAEs 
plus procedures for completing and transmitting SAE reports to GSK are provided in 
Section 12.3.
 
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7.4.1.2.
Method of Detecting AEs and SAEs
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended 
and non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrence.  Appropriate questions include:

“How are you feeling?” 

“Have you had any (other) medical problems since your last visit/contact?”

“Have you taken any new medicines, other than those provided in this study, 
since your last visit/contact?
7.4.1.3.
Follow-up of AEs and SAEs
After the initial AE/SAE report, the investigator is required to proactively follow each 
subject at subsequent visits/contacts.  All SAEs, and non-serious AEs of special interest 
(as defined in Section 4.6.1) will be followed until resolution, until the condition 
stabilizes, until the event is otherwise explained, or until the subject is lost to follow up 
(as defined in Section 5.4). Further information on follow up procedures is given in Study 
Assessments and Procedures.
7.4.1.4.
Regulatory Reporting Requirements for SAEs
Prompt notification by the investigator to GSK of SAEs related to study treatment (even 
for non- interventional post-marketing studies) is essential so that legal obligations and 
ethical responsibilities towards the safety of subjects and the safety of a product under 
clinical investigation are met. 
GSK has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of a product under clinical investigation.  GSK will 
comply with country specific regulatory requirements relating to safety reporting to the 
regulatory authority, Institutional Review Board (IRB)/Independent Ethics Committee 
(IEC) and investigators.
Investigator safety reports are prepared for suspected unexpected serious adverse 
reactions according to local regulatory requirements and GSK policy and are forwarded 
to investigators as necessary.
An investigator who receives an investigator safety report describing a SAE(s) or other 
specific safety information (e.g., summary or listing of SAEs) from GSK will file it with 
the IB and will notify the IRB/IEC, if appropriate according to local requirements.
7.4.2.
Physical Exams

A complete physical examination will include, at a minimum, assessment of the 
Cardiovascular, Respiratory, Gastrointestinal and Neurological systems. Height 
and weight will also be measured and recorded. 

A brief physical examination will include, at a minimum assessments of the skin, 
lungs, cardiovascular system, and abdomen (liver and spleen).
 
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
Investigators should pay special attention to clinical signs related to previous 
serious illnesses
7.4.3.
Vital Signs

Vital signs will be measured in semi-supine position after 5 minutes rest and will 
include temperature, systolic and diastolic blood pressure and pulse rate and 
respiratory rate. 

Three readings of blood pressure and pulse rate will be taken at Screening and 
baseline (pre-dose). All subsequent assessments will be single measures, unless 
the subjects blood pressure or pulse rate has changed from baseline by >15% and 
then 2 further readings should be taken and recorded:

For triplicate readings:

All 3 readings will be recorded in the CRF

First reading should be rejected

Second and third readings should be averaged to give the measurement to 
be used and this will also be recorded in the CRF
7.4.4.
Electrocardiogram (ECG)

ECGs will be measured in semi-supine position after 5 minutes rest

Triplicate 12-lead ECGs will be obtained at Screening and baseline (predose). All 
subsequent assessments will be a single measures, unless withdrawal criteria are 
meet and in which case 2 further readings should be taken to confirm if 
withdrawal is required . An ECG machine that automatically calculates the heart 
rate and measures PR, QRS, QT, and QTc.  Refer to Section 5.4.2 for QTc 
withdrawal criteria and additional QTc readings that may be necessary.

24 hour continuous cardiac telemetry (Holter) will be performed at Screening.  
Full disclosures will be reviewed in detail and the review maintained as part of the 
subject’s source documents
7.4.5.
Clinical Safety Laboratory Assessments
All protocol required laboratory assessments, as defined in Table 8, must be conducted 
in accordance with the Laboratory Manual, and Protocol Time and Events Schedule.
Laboratory requisition forms must be completed and samples must be clearly labelled 
with the subject number, protocol number, site/centre number, and visit dat.  Reference 
ranges for all safety parameters will be provided to the site by the laboratory responsible 
for the assessments.
If additional non-protocol specified laboratory assessments are performed at the 
institution’s local laboratory and result in a change in subject management or are 
considered clinically significant by the investigator (e.g., SAE or AE or dose 
modification) the results must be recorded in the CRF.
 
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Refer to the SRM for appropriate processing and handling of samples to avoid duplicate 
and/or additional blood draws.
All study-required laboratory assessments will be performed by a local laboratory.
Haematology, clinical chemistry, urinalysis and additional parameters to be tested are 
listed in Table 8. 
Table 8
Protocol Required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Haematology
Platelet Count
RBC Indices:
WBC count with Differential:
RBC Count
MCV
Neutrophils
Hemoglobin
MCH
Lymphocytes
Hematocrit
MCHC
Monocytes
Eosinophils
Basophils
Clinical 
Chemistry 1
BUN
Potassium
AST (SGOT)
Total and direct bilirubin
Creatinine
Sodium
ALT (SGPT)
Total Protein
Glucose 
Calcium
Alkaline phosphatise
Albumin
Routine 
Urinalysis3
As a minimum, but not limited to the following tests and dependent on 
standard urinalysis dipstick used:

Specific gravity

pH, glucose, protein, blood and ketones by dipstick

Microscopic examination (if blood or protein is abnormal)
Other 
Screening 
Tests

HIV

Hepatitis B (HBsAg)

Hepatitis C (Hep C antibody)

Alcohol and drug screen (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines)

Serum  or urine hCG Pregnancy test2
NOTES :
1.
Details of Liver Chemistry Stopping Criteria and Required Actions and Follow-Up 
Assessments after liver stopping or monitoring event are given in Section 5.4.1 and 
Appendix 2
2.
Local urine testing will be standard for the protocol unless serum testing is required 
by local regulation or ethics committee.
3.
Routine Urinalysis results will not be databased, unless a result is out of range and 
clinically significant then it would be captured as an AE  
 
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All laboratory tests with values that are considered clinically significantly abnormal 
during participation in the study or within7 days after the last dose of study treatment 
should be repeated until the values return to normal or baseline.  If such values do not 
return to normal within a period judged reasonable by the investigator, the etiology
should be identified and the sponsor notified.
8.
DATA MANAGEMENT

Data will be double-entered into a clinical database management system 
(ClinPlus Version 3.3).

Management of clinical data will be performed in accordance with applicable 
HMR standards and data cleaning procedures to ensure the integrity of the data, 
e.g., removing errors and inconsistencies in the data.

Adverse events and concomitant medications terms will be coded using 
MedDRA (Medical Dictionary for Regulatory Activities) and an internal 
validated medication dictionary, GSKDrug.  

Original CRFs will be retained by GSK, while HMR will retain a copy. Subject 
initials will not be collected or transmitted to GSK according to GSK policy 
9.
STATISTICAL CONSIDERATIONS AND DATA 
ANALYSES
9.1.
Hypotheses
No formal hypothesis will be tested for study Part 1 and Part 2. An estimation approach 
will be used to (i) estimate the bioavailability of the FDC formulations relative to the 
monotherapies taken concurrently, (ii) for each primary pharmacokinetic endpoint, point 
estimates and corresponding 90% confidence intervals will be constructed for the ratio of 
the geometric mean of the test treatment (FDC formulations) to the geometric mean of 
the reference treatment (co-administration of ambrisentan and tadalafil), (test)/ 
(reference).  The objective of Part 2 is to assess whether differences in granulation size 
impact the pharmacokinetics of ambrisentan and tadalafil; the estimation approach for 
bioavailability is therefore applicable to Part 2.
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 10 
mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 10 mg & 
tadalafil 40 mg taken concurrently (reference) in healthy subjects in both the fed and 
fasted states. The null hypothesis is that the true ratio of the geometric mean of the test 
treatment to the geometric mean of the reference treatment, (test)/(reference), for each 
primary PK endpoint, is either less than 0.80 or greater than 1.25.  The alternate 
hypothesis is that the true ratio of the test treatment geometric mean to the reference 
treatment geometric mean is at least 0.80 and no greater than 1.25.  Symbolically, this is 
expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
 
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i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e., treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test (TOST) 
procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be used to test 
this set of hypotheses. This is equivalent to requiring that a 90% interval for the true ratio 
of test to reference geometric means falls entirely within the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test (1) the bioequivalence of 
a FDC of ambrisentan 5 mg + tadalafil 40 mg (test) relative to reference monotherapies 
of ambrisentan 5 mg & tadalafil 40 mg taken concurrently (reference) and (2) the 
bioequivalence of a FDC of ambrisentan 5 mg + tadalafil 20 mg (test) relative to 
reference monotherapies of ambrisentan 5 mg & tadalafil 20 mg taken concurrently 
(reference), in healthy subjects in the fasted states. Therefore, the analysis approach for 
bioequivalence for Part 3A is applicable to Part 3B.
9.2.
Sample Size Considerations
9.2.1.
Sample Size Assumptions
For Part 1 and Part 2 of the study, the sample size assumptions are based on previously 
reported estimates of within subject CV for AUC(0-∞) and Cmax for ambrisentan (GS-US-
300-0112, 2008) and tadalafil (Forgue, 2005). Table 9 summarizes the estimates of 
within subject CV for the primary endpoints AUC (0-∞) and Cmax.
Table 9
Estimates of within subject CV for the primary end points AUC (0-∞) 
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
For Part 3A of the study, Table 10summarized the observed estimates of between and 
within subject CV and observed ratio of geometric means for the primary endpoints 
AUC(0-t) and Cmax from Part 1 of the study. 
 
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Table 10
Estimates of within subject CV for the primary end points AUC (0-∞)
and Cmax for ambrisentan and tadalafil from part 1 of the study
CVw: within subject CV
ambrisentan 
tadalafil 
Cmax
22.42%
14.53%
AUC (0-∞)
8.12%
10.32%
The largest of the within subject CV estimates 22.42% translates to a standard deviation
(SD) of 0.221 on the natural log scale.  Based on this SD, a sample size of 23 statistically 
evaluable subjects will have 90% power to demonstrate bioequivalence. This calculation 
assumes:

a true ratio of 1,

the within-subject variability from the current study will not be larger than that 
used in the sample size calculations,

data are log-normally distributed, and each t-test is made at the 5% level.
For Part 3B of the study, same assumptions are adopted as Part 3A. Thus, a sample size 
of 23 statistically evaluable subjects will have 90% power to demonstrate bioequivalence.  
However, if the within subject CVs obtained after Part 2 are significantly higher than the 
ones in Part 1 (shown in Table 10), a sample size re-esimation may be conducted to 
adjust the sample size for Part 3A and 3B to ensure the study maintain enough power to 
demonstrate bioequivalence. 
9.2.2.
Sample Size Sensitivity
For Part 1 and Part 2, assuming the within subject CV differs from 22% and the number 
of statistically evaluable subjects is lower than 20, then Table 11 shows the precision (the 
half width of the 90% confidence interval) for a range of evaluable subjects.
 
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Table 11
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
For Part 3, based on the observed data from previous studies, the ratio of geometric mean 
for ambrisentan Cmax is unlikely to be 1. The effects on the power of declaring 
bioequivalence in the face of a shift in the expected ratio of the geometric means were 
examined.
If, under all other assumptions outlined above, the actual ratio of geometric means for 
ambrisentan Cmax is 0.97, then 26 evaluable subjects are needed to have 90% power to 
conclude bioequivalence.
With 26 evaluable subjects, the current study design has at least 90% power to conclude 
bioequivalance (with both Cmax and AUC for ambrisentan and tadalafil).
9.2.3.
Sample Size Re-estimation or Adjustment
No sample size re-estimation will be performed.  However, if the estimation of the within 
subject CV from Part 2 of the study is much larger than the within subject CV from Part 1 
of the study, the sample size for Part3 may be re-evaluated. 
9.3.
Data Analysis Considerations
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
sampling times recorded during the study although supplementary analayis will be 
available based on the nominal times.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-life 
(t1/2).  
 
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Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model.
9.3.1.
Analysis Populations
9.3.1.1.
Safety Population
All subjects enrolled into the study who have received at least one dose of investigational 
product will be included in the Safety Population.
9.3.1.2.
Pharmacokinetic Concentration Population
The PK Concentration Population will include all subjects for whom at least one PK 
sample was obtained and analyzed.
9.3.1.3.
Pharmacokinetic Parameter Population
For each PK parameter, the PK Parameter Population will include all subjects who 
provide PK parameter data.
9.3.2.
Interim Analysis
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in Part 1 and Part 2 of the study in order to direct development of the FDC 
for later parts of the study.  Headline results based on statistical analysis using 
preliminary pharmacokinetic data with nominal time may be produced when 80% and 
100% subjects complete Part 1 and Part 2 of the study to assist development of FDCs. 
Treatment and period information from the crossover design may be used in the analysis; 
sequence information may also be included depending on availability. 
9.4.
Key Elements of Analysis Plan
9.4.1.
Pharmacokinetic Analyses
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
 
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treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D. 
Data will be listed and summarized according to GlaxoSmithKline reporting standards, 
where applicable.  Listings will be sorted by subject, period, noting treatment.  
Summaries will be presented by treatment.  Unless stated otherwise, descriptive 
summaries will include n, mean, standard deviation (SD), coefficient of variation (CV), 
median, minimum, and maximum for continuous variables, n and percent for categorical 
variables and geometric mean, 95% confidence interval (CI), and the between-subject CV 
(CVb) based on geometric mean for the log-transformed PK parameters.
Version 9.3 (or higher) of the SAS system will be used for statistical analysis of the data
as well as to generate tables, figures, and listings.
Any deviation(s) from the original analyses planned in the protocol will be reported in the 
Reporting and Analysis Plan (RAP) and/or in the Clinical Pharmacology Study Report.
The PK parameters, AUC(0-), AUC(0-t), Cmax, and t½ will be transformed using natural 
logarithms. Missing PK parameters will not be imputed.
All data from withdrawn subjects will be listed.
No adjustment for multiple tests or comparisons is planned.
Actual visit day will be used for safety assessments. For the assessment of 
bioequivalence, both actual time of sampling, and nominal (planned) sampling time, will 
be used to estimate PK parameters at the end of the study. PK parameters estimation 
based on nominal sampling time will be used for generating headline results.Following 
loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations and reference will 
be separately analyzed using a mixed effects model with fixed effect terms for period, 
sequence and treatment, and subject as random effect term (i.e. ANOVA method using 
period, sequence and treatment as fixed effect, and subject as random effect) Point 
estimates and their associated 90% confidence intervals will be calculated for the 
differences: F1 - R, F2 - R, F3 -R, F4 - R for Part 1; FG1-R, FG2-R, FG3-R for Part 2; 
X1-R1, X2-R2 for Part 3A; and Y-R3 for Part 3B. The point estimates and their 
associated 90% confidence intervals will then be back-transformed to provide point 
estimates and 90% confidence intervals for the ratios, F1:R, F2:R, F3:R, F4:R; for Part 1; 
FG1:R, FG2:R, FG3:R for Part 2; X1:R1,X2:R2 for Part3A; and Y:R3 for Part 3B.
9.4.2.
Safety Analyses
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
 
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10.
STUDY GOVERNANCE CONSIDERATIONS
10.1.
Posting of Information on Publicly Available Clinical Trial 
Registers
Study information from this protocol will be posted on publicly available clinical trial 
registers before enrollment of subjects begins.
10.2.
Regulatory and Ethical Considerations, Including the 
Informed Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.
The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent

Investigator reporting requirements (e.g. reporting of AEs/SAEs/protocol 
deviations to IRB/IEC)

GSK will provide full details of the above procedures, either verbally, in writing, 
or both.

Signed informed consent must be obtained for each subject prior to participation 
in the study

The IEC/IRB, and where applicable the regulatory authority, approve the clinical 
protocol and all optional assessments, including genetic research.  

Optional assessments (including those in a separate protocol and/or under separate 
informed consent) and the clinical protocol should be concurrently submitted for 
approval unless regulation requires separate submission.  

Approval of the optional assessments may occur after approval is granted for the 
clinical protocol where required by regulatory authorities.  In this situation, 
written approval of the clinical protocol should state that approval of optional 
assessments is being deferred and the study, with the exception of the optional 
assessments, can be initiated.
 
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10.3.
Quality Control (Study Monitoring)

In accordance with applicable regulations including GCP, and GSK procedures, 
GSK monitors will contact the site prior to the start of the study to review with the 
site staff the protocol, study requirements, and their responsibilities to satisfy 
regulatory, ethical, and GSK requirements.  

When reviewing data collection procedures, the discussion will also include 
identification, agreement and documentation of data items for which the CRF will 
serve as the source document.
GSK will monitor the study and site activity to verify that the:

Data are authentic, accurate, and complete.

Safety and rights of subjects are being protected.

Study is conducted in accordance with the currently approved protocol and any 
other study agreements, GCP, and all applicable regulatory requirements.
The investigator and the head of the medical institution (where applicable) agrees to 
allow the monitor direct access to all relevant documents
10.4.
Quality Assurance

To ensure compliance with GCP and all applicable regulatory requirements, GSK 
may conduct a quality assurance assessment and/or audit of the site records, and 
the regulatory agencies may conduct a regulatory inspection at any time during or 
after completion of the study. 

In the event of an assessment, audit or inspection, the investigator (and institution) 
must agree to grant the advisor(s), auditor(s) and inspector(s) direct access to all 
relevant documents and to allocate their time and the time of their staff to discuss 
the conduct of the study, any findings/relevant issues and to implement any 
corrective and/or preventative actions to address any findings/issues identified.
10.5.
Study and Site Closure

Upon completion or premature discontinuation of the study, the GSK monitor will 
conduct site closure activities with the investigator or site staff, as appropriate, in 
accordance with applicable regulations including GCP, and GSK Standard 
Operating Procedures.

GSK reserves the right to temporarily suspend or prematurely discontinue this 
study at any time for reasons including, but not limited to, safety or ethical issues 
or severe non-compliance. For multicenter studies, this can occur at one or more 
or at all sites.  

If GSK determines such action is needed, GSK will discuss the reasons for taking 
such action with the investigator or the head of the medical institution (where 
applicable).  When feasible, GSK will provide advance notification to the 
investigator or the head of the medical institution, where applicable, of the 
impending action.
 
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
If the study is suspended or prematurely discontinued for safety reasons, GSK will 
promptly inform all investigators, heads of the medical institutions (where 
applicable) and/or institution(s) conducting the study. GSK will also promptly 
inform the relevant regulatory authorities of the suspension or premature 
discontinuation of the study and the reason(s) for the action.  

If required by applicable regulations, the investigator or the head of the medical 
institution (where applicable) must inform the IRB/IEC promptly and provide the 
reason for the suspension or premature discontinuation.
10.6.
Records Retention

Following closure of the study, the investigator or the head of the medical 
institution (where applicable) must maintain all site study records (except for 
those required by local regulations to be maintained elsewhere), in a safe and 
secure location.  

The records must be maintained to allow easy and timely retrieval, when needed 
(e.g., for a GSK audit or regulatory inspection) and must be available for review 
in conjunction with assessment of the facility, supporting systems, and relevant 
site staff.  

Where permitted by local laws/regulations or institutional policy, some or all of 
these records can be maintained in a format other than hard copy (e.g., microfiche, 
scanned, electronic); however, caution needs to be exercised before such action is 
taken.  

The investigator must ensure that all reproductions are legible and are a true and 
accurate copy of the original and meet accessibility and retrieval standards, 
including re-generating a hard copy, if required.  Furthermore, the investigator 
must ensure there is an acceptable back-up of these reproductions and that an 
acceptable quality control process exists for making these reproductions.

GSK will inform the investigator of the time period for retaining these records to 
comply with all applicable regulatory requirements.  The minimum retention time 
will meet the strictest standard applicable to that site for the study, as dictated by 
any institutional requirements or local laws or regulations, GSK 
standards/procedures, and/or institutional requirements.  

The investigator must notify GSK of any changes in the archival arrangements, 
including, but not limited to, archival at an off-site facility or transfer of 
ownership of the records in the event the investigator is no longer associated with 
the site.
10.7.
Provision of Study Results to Investigators, Posting of 
Information on Publically Available Clinical Trials Registers 
and Publication
Where required by applicable regulatory requirements, an investigator signatory will be 
identified for the approval of the clinical study report.  The investigator will be provided 
reasonable access to statistical tables, figures, and relevant reports and will have the 
 
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opportunity to review the complete study results at a GSK site or other mutually-
agreeable location.
GSK will also provide the investigator with the full summary of the study results.  The 
investigator is encouraged to share the summary results with the study subjects, as 
appropriate.
GSK will provide the investigator with the randomization codes for their site only after 
completion of the full statistical analysis.
The procedures and timing for public disclosure of the results summary and for 
development of a manuscript for publication will be in accordance with GSK Policy.
A manuscript will be progressed for publication in the scientific literature if the results 
provide important scientific or medical knowledge.
 
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11.
REFERENCES
Adcirca: EMA Summary of product Characteristics, Eli Lily, last updated 29-Sep-2015 at 
time of publication
EMEA guidelines on clinical development of fixed dose combination medicinal products. 
CHMP/EWP/240/95 Rev.1. 2008
EMEA guidelines on the investigation of bioequivalence. CPMP/EWP/QWP/1401/98 
Rev. 1/ Corr **. 2010
Forgue S, Patterson B, Bedding A, et al. Tadalafil pharmacokinetics in healthy subjects. 
British Journal of Clinical Pharmacology. 20-Dec-2005
Galiè N et al. Initial Use of Ambrisentan plus Tadalafil in Pulmonary Arterial 
Hypertension. N Engl J Med 2015a;373:834-44.
Galiè N, Humbert M, Vachiery J.L et al. 2015 ESC/ERS Guidelines for the diagnosis and 
treatment of pulmonary hypertension. European Heart Journal Advance Access, 
published August 29, 2015b
Ghofrani AH, Galiè N, Grimminger F, et al. N Engl J Med 2013;369:330-40. Riociguat 
for the Treatment of Pulmonary Arterial Hypertension
Gilead Clinical Study report GS-US-300-0112: A Phase 1, Open-Label, Randomized, 
Two-Period, Crossover Study to Assess the Pharmacokinetics of Ambrisentan Alone and 
in the Presence of Multiple Doses of Tadalafil, and to assess the Pharmacokinetics of 
Tadalafil Alone and in the Presence of Multiple Doses of Ambrisentan in Healthy Adult 
Volunteers. Version 1.0, dated 27-June-2008
Gilead Investigator Brochure, Letaris, Volibris, Fifteenth edition: 06 April 2015
GSK Clinical Study Report for study AMB112565: AMBITION: A Randomized, 
Multicenter Study of First-Line Ambrisentan and Tadalafil Combination Therapy in 
Subjects with Pulmonary Arterial Hypertension. GSK document 2014N193963_00, dated 
13-Nov-14
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(a): 24 Table 3-2
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(b): 28.
Liang F, Yang S, Yao L, Belardinelli L, Shryock J. Hypertension. 2012 Mar;59(3):705-
11. Ambrisentan and tadalafil synergistically relax endothelin-induced contraction of rat 
pulmonary arteries.
Pulido T, Adzerikho I, Channick RN, et al. N Engl J Med. 2013 Aug 29;369(9):809-18. 
Macitentan and Morbidity and Mortality in Pulmonary Arterial Hypertension
 
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Schuirmann DJ. A comparison of the two one-sided tests procedure and the power 
approach for assessing the equivalence of average bioavailability, J Pharmacokin. 
Biopharm. 1987:15:657–680. 
Spence R, Mandagere A, Harrison B, et al.  J Pharm Sci. 2009 Dec;98(12):4962-74. No 
clinically relevant pharmacokinetic and safety interactions of ambrisentan in combination 
with tadalafil in healthy volunteers. 
Volibris: EMA Summary of Product Characteristics, GSK, variation II/41, at time of 
publication, 2015.
 
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12.
APPENDICES
12.1.
Appendix 1: Abbreviations and Trademarks
Abbreviations
AE
Adverse Event
API
Active Pharmaceutical Ingredient
AUC
Area Under Curve
BE
Bioequivalence
CDISC
Clinical Data Interchange Standards Consortium
cGMP
Cyclic guanosine monophosphate
CI
Confidence Interval
Cmax
Maximum concentration
CV
Coefficient of variance 
CVw
Coefficient of variance within subject
EMA
European Medicines Agency 
ERA
Endothelin Receptor Antagonists
ERS
European Respiratory Society 
ESC
European Society of Cardiology
ET-1
Endothelin Receptor - 1
EU
European Union
FDA
Federal Drug Agency
FDC
Fixed Dose Combination
NO
Nitrous Oxide
PDE-5
Phosphodiesterase type 5
SAE
Serious Adverse Events
SRM
Study Reference Manual
t1/2
Half Life
tmax
Time to maximum concentration
WHO
World Health Organisation
Trademark Information
Trademarks of the GlaxoSmithKline 
group of companies
Trademarks not owned by the 
GlaxoSmithKline group of companies
NONE
Adcirca
WinNonlin
 
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12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments
Phase I Liver chemistry stopping criteria have been designed to assure subject safety and 
to evaluate liver event aetiology (in alignment with the FDA premarketing clinical liver 
safety guidance).
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf.
Phase I liver chemistry stopping criteria and required follow up assessments
Liver Chemistry Stopping Criteria – Liver Stopping Event
ALT-absolute
ALT≥3xULN
If  ALT≥3xULN  AND bilirubin1,2 2xULN (>35% direct bilirubin) or  INR >1.5, 
Report as an SAE.
See additional Actions and Follow-Up Assessments listed below
Required Actions and Follow up Assessments following Liver Stopping Event
Actions
Follow-Up Assessments

Immediately discontinue study treatment 

Report the event to GSK within 24 hours

Complete the liver event CRF, and complete 
an SAE data collection tool if the event also 
meets the criteria for an SAE2

Perform liver event follow up assessments 

Monitor the subject until liver chemistries 
resolve, stabilise, or return to within baseline 
(see MONITORING below)
MONITORING:
If ALT≥3xULN AND bilirubin 2xULN  or INR 
>1.5

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within  24 
hrs

Monitor subjects twice weekly until liver 
chemistries resolve, stabilise or return to 
within baseline

Viral hepatitis serology3

Blood sample for pharmacokinetic (PK) 
analysis, obtained within 96hrs of last dose

Serum creatine phosphokinase (CPK) and 
lactate dehydrogenase (LDH).

Fractionate bilirubin, if total 
bilirubin2xULN

Obtain complete blood count with 
differential to assess eosinophilia

Record the appearance or worsening of 
clinical symptoms of liver injury, or 
hypersensitivity, on the AE report form

Record use of concomitant medications on 
the concomitant medications report form 
including acetaminophen, herbal remedies, 
other over the counter medications.

Record alcohol use on the liver event 
alcohol intake case report form
If ALT≥3xULN AND bilirubin 2xULN  or 
 
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Liver Chemistry Stopping Criteria – Liver Stopping Event

A specialist or hepatology consultation is 
recommended
If ALT≥3xULN AND bilirubin < 2xULN  and 
INR ≤1.5:

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within 24-
72 hrs

Monitor subjects weekly until liver 
chemistries resolve, stabilize or return to 
within baseline
INR >1.5:

Anti-nuclear antibody, anti-smooth muscle 
antibody, Type 1 anti-liver kidney 
microsomal antibodies, and quantitative 
total immunoglobulin G (IgG or gamma 
globulins).

Serum acetaminophen adduct HPLC assay 
(quantifies potential acetaminophen 
contribution to liver injury in subjects with 
definite or likely acetaminophen use in the 
preceding week [James, 2009].

Liver imaging (ultrasound, magnetic 
resonance, or computerised tomography) 
and /or liver biopsy to evaluate liver 
disease; complete Liver Imaging and/or 
Liver Biopsy CRF forms.
1.
Serum bilirubin fractionation should be performed if testing is available. If serum bilirubin fractionation is not 
immediately available, discontinue study treatment for that subject if ALT 3xULN and bilirubin 2xULN. 
Additionally, if serum bilirubin fractionation testing is unavailable, record presence of detectable urinary bilirubin on 
dipstick, indicating direct bilirubin elevations and suggesting liver injury. 
2.
All events of ALT 3xULN and bilirubin 2xULN (>35% direct bilirubin) or ALT 3xULN and INR>1.5, if INR 
measured, which may indicate severe liver injury (possible ‘Hy’s Law’), must be reported as an SAE (excluding 
studies of hepatic impairment or cirrhosis); INR measurement is not required and the threshold value stated will 
not apply to subjects receiving anticoagulants
3.
Includes: Hepatitis A IgM antibody; Hepatitis B surface antigen and Hepatitis B Core Antibody (IgM); Hepatitis C 
RNA; Cytomegalovirus IgM antibody;  Epstein-Barr viral capsid antigen IgM antibody (or if unavailable, obtain 
heterophile antibody or monospot testing);  Hepatitis E IgM antibody 
4.
PK sample may not be required for subjects known to be receiving placebo or non-GSK comparator treatments.  
Record the date/time of the PK blood sample draw and the date/time of the last dose of study treatment prior to 
blood sample draw on the CRF. If the date or time of the last dose is unclear, provide the subject’s best 
approximation. If the date/time of the last dose cannot be approximated OR a PK sample cannot be collected in 
the time period indicated above, do not obtain a PK sample. Instructions for sample handling and shipping are in 
the SRM. 
References
James LP, Letzig L, Simpson PM, Capparelli E, Roberts DW, Hinson JA, Davern TJ, Lee 
WM. Pharmacokinetics of Acetaminophen-Adduct in Adults with Acetaminophen 
Overdose and Acute Liver Failure. Drug Metab Dispos 2009; 37:1779-1784.
 
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12.3.
Appendix 3: Definition of and Procedures for Recording,
Evaluating, Follow-Up and Reporting of Adverse Events
12.3.1.
Definition of Adverse Events
Adverse Event Definition:

An AE is any untoward medical occurrence in a patient or clinical investigation 
subject, temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product.

NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal product.
Events meeting AE definition include:

Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (e.g., ECGs, radiological scans, vital signs 
measurements), including those that worsen from baseline, and felt to be clinically 
significant in the medical and scientific judgement of the investigator.

Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

New conditions detected or diagnosed after study treatment administration even 
though it may have been present prior to the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected interaction.

Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication (overdose per se will not be reported as an 
AE/SAE unless this is an intentional overdose taken with possible suicidal/self-
harming intent.  This should be reported regardless of sequelae).
Events NOT meeting definition of an AE include:

Any clinically significant abnormal laboratory findings or other abnormal safety 
assessments which are associated with the underlying disease, unless judged by the 
investigator to be more severe than expected for the subject’s condition.

The disease/disorder being studied or expected progression, signs, or symptoms of 
the disease/disorder being studied, unless more severe than expected for the subject’s 
condition.

Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that 
leads to the procedure is an AE.

Situations where an untoward medical occurrence did not occur (social and/or 
 
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convenience admission to a hospital).

Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen.
12.3.2.
Definition of Serious Adverse Events
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, 
death due to progression of disease, etc).
Serious Adverse Event (SAE) is defined as any untoward medical occurrence that, 
at any dose:
a.
Results in death
b.
Is life-threatening
NOTE:
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
subject was at risk of death at the time of the event.  It does not refer to an event, which 
hypothetically might have caused death, if it were more severe.
c.
Requires hospitalization or prolongation of existing hospitalization
NOTE:

In general, hospitalization signifies that the subject has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for 
observation and/or treatment that would not have been appropriate in the physician’s 
office or out-patient setting.  Complications that occur during hospitalization are 
AEs.  If a complication prolongs hospitalization or fulfills any other serious criteria, 
the event is serious.  When in doubt as to whether “hospitalization” occurred or was 
necessary, the AE should be considered serious.

Hospitalization for elective treatment of a pre-existing condition that did not worsen 
from baseline is not considered an AE.
d.
Results in disability/incapacity
NOTE:

The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.

This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, 
and accidental trauma (e.g. sprained ankle) which may interfere or prevent everyday 
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e.
Is a congenital anomaly/birth defect
f.
Other situations:

Medical or scientific judgment should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be 
immediately life-threatening or result in death or hospitalization but may jeopardize 
the subject or may require medical or surgical intervention to prevent one of the 
other outcomes listed in the above definition.  These should also be considered 
serious.

Examples of such events are invasive or malignant cancers, intensive treatment in an 
emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse
g.
Is associated with liver injury and impaired liver function defined as:

ALT 3xULN and total bilirubin* 2xULN (>35% direct), or

ALT 3xULN and INR** > 1.5.
* Serum bilirubin fractionation should be performed if testing is available; if unavailable, 
measure urinary bilirubin via dipstick.  If fractionation is unavailable and ALT 3xULN 
and total bilirubin 2xULN, then the event is still to be reported as an SAE.
** INR testing not required per protocol and the threshold value does not apply to 
subjects receiving anticoagulants.  If INR measurement is obtained, the value is to be 
recorded on the SAE form.

Refer to Section 12.2 for the required liver chemistry follow up instructions
12.3.3.
Definition of Cardiovascular Events
Cardiovascular Events (CV) Definition:
Investigators will be required to fill out the specific CV event page of the CRF for the 
following AEs and SAEs:

Myocardial infarction/unstable angina

Congestive heart failure

Arrhythmias

Valvulopathy

Pulmonary hypertension

Cerebrovascular events/stroke and transient ischemic attack

Peripheral arterial thromboembolism

Deep venous thrombosis/pulmonary embolism
 
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
Revascularization
12.3.4.
Recording of AEs and SAEs
AEs and SAE Recording:

When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) 
relative to the event.

The investigator will then record all relevant information regarding an AE/SAE in 
the CRF

It is not acceptable for the investigator to send photocopies of the subject’s medical 
records to GSK in lieu of completion of the GSK, AE/SAE CRF page.

There may be instances when copies of medical records for certain cases are 
requested by GSK.  In this instance, all subject identifiers, with the exception of the 
subject number, will be blinded on the copies of the medical records prior to 
submission of to GSK.

The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information.  In such cases, the diagnosis will be 
documented as the AE/SAE and not the individual signs/symptoms.

Subject-completed Value Evidence and Outcomes questionnaires and the collection 
of AE data are independent components of the study.

Responses to each question in the Value Evidence and Outcomes questionnaire will 
be treated in accordance with standard scoring and statistical procedures detailed by 
the scale’s developer.

The use of a single question from a multidimensional health survey to designate a 
cause-effect relationship to an AE is inappropriate.
12.3.5.
Evaluating AEs and SAEs
Assessment of Intensity
The investigator will make an assessment of intensity for each AE and SAE reported 
during the study and will assign it to one of the following categories:

Mild:  An event that is easily tolerated by the subject, causing minimal discomfort 
and not interfering with everyday activities.

Moderate:  An event that is sufficiently discomforting to interfere with normal 
everyday activities

Severe:  An event that prevents normal everyday activities. - an AE that is assessed 
as severe will not be confused with an SAE. Severity is a category utilized for rating 
 
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the intensity of an event; and both AEs and SAEs can be assessed as severe.

An event is defined as ‘serious’ when it meets at least one of the pre-defined 
outcomes as described in the definition of an SAE.
Assessment of Causality

The investigator is obligated to assess the relationship between study treatment and 
the occurrence of each AE/SAE.

A "reasonable possibility" is meant to convey that there are facts/evidence or 
arguments to suggest a causal relationship, rather than a relationship cannot be ruled 
out.

The investigator will use clinical judgment to determine the relationship.

Alternative causes, such as natural history of the underlying diseases, concomitant 
therapy, other risk factors, and the temporal relationship of the event to the study 
treatment will be considered and investigated.

The investigator will also consult the Investigator Brochure (IB) and/or Product 
Information, for marketed products, in the determination of his/her assessment.

For each AE/SAE the investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality.

There may be situations when an SAE has occurred and the investigator has minimal 
information to include in the initial report to GSK.  However, it is very important 
that the investigator always make an assessment of causality for every event 
prior to the initial transmission of the SAE data to GSK.

The investigator may change his/her opinion of causality in light of follow up
information, amending the SAE data collection tool accordingly.

The causality assessment is one of the criteria used when determining regulatory 
reporting requirements.
Follow up of AEs and SAEs

The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as may be indicated or as requested by GSK to 
elucidate as fully as possible the nature and/or causality of the AE or SAE.

The investigator is obligated to assist.  This may include additional laboratory tests 
or investigations, histopathological examinations or consultation with other health 
care professionals.

If a subject dies during participation in the study or during a recognized follow up 
period, the investigator will provide GSK with a copy of any post-mortem findings, 
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
New or updated information will be recorded in the originally completed CRF.

The investigator will submit any updated SAE data to GSK within the designated 
reporting time frames.
12.3.6.
Reporting of SAEs to GSK
SAE reporting to GSK via paper CRF

Facsimile transmission of the SAE paper CRF is the preferred method to transmit 
this information to the Medical Monitor or the SAE coordinator

In rare circumstances and in the absence of facsimile equipment, notification by 
telephone is acceptable, with a copy of the SAE data collection tool sent by 
overnight mail

Initial notification via the telephone does not replace the need for the investigator to 
complete and sign the SAE CRF pages within the designated reporting time frames.

Contacts for SAE receipt can be found at this beginning of the protocol on the 
Sponsor/Medical Monitor Contact Information page.
 
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12.4.
Appendix 4 High Fat Meal Content

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
The standard high-fat meal will be the meal suggested by the EMEA guidelines on the 
investigation of bioequivalence 2010. The meal should be a high-fat (approximately 50 
percent of total caloric content of the meal) and high-calorie (approximately 800 to 1000 
kcal) meal. This test meal should derive approximately 150, 250, and 500-600 kcal from 
protein, carbohydrate, and fat, respectively.(EMEA, 2010)
 
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12.5.
Appendix 5: Protocol Changes
12.5.1.
Protocol changes for Amendment 01 (14-Apr-2016) from Original 
Protocol (16-Dec-2015)
The purpose of this amendment is to clarify the hypotension withdrawal criteria in 
section 5.4.3, address changes to GSK medical monitor and minor administrative 
changes.
Summary of Changes
Section 1 PROTOCOL SYNOPSIS FOR STUDY 201964
AND
Section 3 OBJECTIVES AND ENDPOINTS
CHANGE FROM
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assed by vital signs, ECG, clinical laboratory 
safety data and review of adverse events
CHANGE TO
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
 
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Section 5.4.3 Hypotension
CHANGE FROM
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 or diastolic <50 confirmed by triplicate reading 
taken up to 5 minutes apart.
CHANGE TO
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 mm HG and diastolic <50 mm HG confirmed 
by triplicate reading taken up to 5 minutes apart and is judged clinically 
significant and symptomatic by the investigator. 
Section 6.3 Treatment Assignment
ADDITION 
Table 6 Treatment Sequence assignments for Part 1, Part 2 and Part 3
Part 2 and 
Part 3 
merged 
together
5 (Two FDCs selected from 
Part 1)
X1 X2 R Y1 Y2
X2 Y1 X1 Y2 R
Y1 Y2 X2 R X1
Y2 R Y1 X1 X2
R X1 Y2 X2 Y1
Y2 Y1 R X2 X1
R Y2 X1 Y1 X2
X1 R X2 Y2 Y1
X2 X1 Y1 R Y2
Y1 X2 Y2 X1 R
1:1:1:1:1:1:1:1:1:1
3 (One FDC selected from 
Part 1)
X1 X2 R
R X2 X1
X2 R X1
F1 R X2
R X1 X2
X2 X1 R
3:3:3:3:3:3 + 2 random
 
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12.5.2.
Protocol changes for Amendment 2 () from Amendment 1(14-Apr-
2016)
Amendment 2 Summary of Changes
The purpose of this substantial amendment is to include bioequivalence assessment in 
Part 3 to replace the optional food effect and the addition of new dosage strengths. 
Furthermore GSK medical monitor contact have changed 
List of Specific Changes
Medical Monitor/SAE Contact Information
Rationale for Change: GSK medical monitor contact have changed
CHANGE FROM
Role
Name
Day Time Phone Number and 
email address
After-hours Phone/Cell/
Pager Number
Fax 
Number
Primary 
Medical 
Monitor
Site to 
scan and 
send to 
email
Secondary 
Monitor
Site to 
scan and 
send to 
email
SAE 
contact  
information 
Site to 
scan and 
send to 
email
CHANGE TO
Role
Name
Day Time Phone Number and 
email address
After-hours 
Phone/Cell/
Pager Number
Fax 
Number
Primary 
Medical 
Monitor
Site to 
scan and 
send to 
email
Secondary 
Medical 
Monitor
Site to 
scan 
and 
send to 
email
 
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Role
Name
Day Time Phone Number and 
email address
After-hours 
Phone/Cell/
Pager Number
Fax 
Number
SAE 
contact  
information 
Site to 
scan 
and 
send to 
email
Section 1 - Protocol Synopsis Overall design
CHANGE FROM:
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects.
All subjects will attend the unit for Screening within 31 days of their first dose. Eligible 
subjects will be randomised to order of treatment in that study parts in which they are 
included and the order in which these treatments are administered will be in accordance 
with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose.  The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
 
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However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast
CHANGE TO: 
This is a single centre, Phase 1, single dose, randomised, open label crossover study in 
healthy volunteers with 3 study parts. Part 1 will include a 5 way cross-over. Part 2 
and Part 3 (A&B) will each include a 4 way cross-over. All subjects will attend the 
unit for Screening within 31 days of their first dose. Eligible subjects will be 
randomised to order of treatment in that study parts in which they are included and 
the order in which these treatments are administered will be in accordance with the 
randomisation schedule.
There will be a minimum washout of 7 days between each dose in Parts 1 and 2. In Part 
3 the washout will be at least 10 days.  A Follow-Up Visit should be completed within 
7-14 days of completion of a subject’s last dose.  The utility of each of the 3 study parts is 
as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
 
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Part 2
This study part will include an evaluation of  the bioavailability, safety and 
tolerability of 3 granulation forms of a single FDC (ambrisentan 10mg + tadalafil 40 
mg) identified from Part 1. These data will be compared to that for the reference of 
the 2 monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 
40 mg). All doses are taken in the fasted state.
Part 3 A
Part 3A of the study is set to establish bioequivalence between the candidate FDC 
from Part 2. This study part will have 4 dose sessions and will assess the 
bioequivalence, in both the fed and fasted state, of the FDC against the reference 
ambrisentan 10 mg + tadalafil 40mg monotherapies in the fed and fasted state.  The 
fed arms of this part will have a standard high fat breakfast. (EMEA, 2010).
Part 3 B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 
40mg and ambrisentan 5 mg + tadalafil 20mg). This study part will have 4 dose 
sessions and will assess the bioequivalence of the two FDC dose strengths against the 
reference ambrisentan 5 mg + tadalafil 40mg and ambrisentan 5 mg + tadalafil 
20mg monotherapies in the fasted state.
Section 1 - Protocol Synopsis Treatment Arms and Duration
CHANGE FROM: 
The proposed treatment arms for each study part are described here; however the 
treatments in Parts 2 and 3 may be changed dependent on the utility and results from the 
previous part.
 
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Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil FDC5 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil FDC6 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil FDC7 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC4 (10mg/40mg)
ambrisentan and tadalafil FDC8 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less FDC treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. The minimum will be approximately 3 weeks and the maximum will be 
approximately 18 weeks.
CHANGE TO:
The proposed treatment arms for each study part are described here
 
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Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)
Part 3A (fed and fasted)
Part 3B (fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 1
(10mg/40mg)
ambrisentan and tadalafil 
FDC ) (10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 2
(10mg/40mg)
ambrisentan and tadalafil 
FDC (10mg/40mg) fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
granulation 3
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg), 
fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
Section 1 - Protocol Synopsis Type and Number of Subjects
CHANGE FROM: 
A approximately of 20 healthy adult subjects will be randomized, to each study part, such 
that at least 16 evaluable subjects complete each part of the study
 
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CHANGE TO: 
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable 
subjects complete each part of the study.  Approximately 33 healthy subjects will be 
randomized to study Part 3A, such that at least 26 evaluable subjects complete part 
3A of the study. An additional 33 healthy subejcts will be randomised to Part 3B, 
such that at least 26 evaluable subjects complete part 3B of the study.
Section 1 - Protocol Synopsis Analysis
CHANGE FROM: 
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and Part 2, (ii) estimate the bioavailability of the formulation(s) of 
the FDC formulations, taken in Part 2, if used for food effect and  Part 3, in the fed state 
relative to the fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
CHANGE TO:
No formal hypothesis will be tested for Part 1 and Part 2 of the study. An estimation 
approach will be used to estimate the bioavailability of the FDC formulations 
relative to the monotherapies taken concurrently.  
For each primary pharmacokinetic endpoint, point estimates and corresponding 
90% confidence intervals will be constructed for the ratio of the geometric mean of 
the test treatment (FDC formulations) to the geometric mean of the reference 
treatment (co-administration of ambrisentan and tadalafil), (test)/ (reference).
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 
10 mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 
10 mg & tadalafil 40 mg taken concurrently (reference) in healthy subjects in both 
the fed and fasted states. The null hypothesis is that the true ratio of the geometric 
mean of the test treatment to the geometric mean of the reference treatment, 
(test)/(reference), for each primary PK endpoint, is either less than 0.80 or 
greater than 1.25.  The alternate hypothesis is that the true ratio of the test 
treatment geometric mean to the reference treatment geometric mean is at least 0.80 
and no greater than 1.25.  Symbolically, this is expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
 
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i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e., treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test 
(TOST) procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be 
used to test this set of hypotheses.  This is equivalent to requiring that a 90% 
interval for the true ratio of test to reference geometric means falls entirely within 
the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test the bioequivalence 
of a FDC of ambrisentan 5 mg + tadalafil 40 mg (test) and  of ambrisentan 5 mg + 
tadalafil 20 mg (test) relative to reference monotherapies of  ambrisentan 5 mg & 
tadalafil 40 mg and of ambrisentan 5 mg & tadalafil 20 mg respectively taken 
concurrently (reference) in healthy subjects in the fasted states. Therefore, the 
analysis approach for bioequivalence for Part 3A is  applicable to Part 3B.
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
Section 2 Introduction
CHANGE FROM
Ambrisentan (E.U. trade name: Volibris), an orally active endothelin receptor antagonists
(ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first approved on 
15 June 2007 in the US and on 21 April 2008 in the European Union (EU) and is 
currently approved in over 50 countries. In the EU, ambrisentan is indicated for treatment 
of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional Class 
(FC) II to III, including use in combination treatment (Volibris, EMA SmPC, 2015 
Section 5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated 
with connective tissue disease
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (EMEA, 2010;Schuirmann, 1987;Adcirca, 2015)A recently completed 
study (Galiè, 2015a) has show that patients with PAH who started initial combination 
therapy with ambrisentan and tadalafil had a significantly lower risk of clinical-failure 
events compared to those that started with ambrisentan or tadalafil monotherapy. 
Ambrisentan has recently received EU approval (20 November 2015) for use in 
combination treatment with tadalafil (Volibris EMA SmPC, 2015, Section 5.1). 
 
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This pilot study will investigate the relative bioavailability of new fixed dose 
combinations (FDC) of ambrisentan and tadalafil, compared to the two monotherapies 
taken concurrently in healthy subjects
CHANGE TO
Ambrisentan (E.U. trade name: Volibris), is an orally active endothelin receptor 
antagonist (ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first 
approved on 15 June 2007 in the US and on 21 April 2008 in the European Union (EU) 
and is currently approved in over 50 countries. In the EU, ambrisentan is indicated for 
treatment of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional 
Class (FC) II to III, including use in combination treatment (Volibris EMA SmPC, 2015 
Section 5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated 
with connective tissue disease.
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (EMEA, 2010;Schuirmann, 1987;Adcirca, 2015).
A recently completed study (Galiè, 2015a) has shown that patients with PAH who started 
initial combination therapy with ambrisentan and tadalafil had a significantly lower risk 
of clinical-failure events compared to those that started with ambrisentan or tadalafil 
monotherapy. Ambrisentan has recently received EU approval (20 November 2015) for 
use in combination treatment with tadalafil (Volibris EMA SmPC, 2015, Section 5.1).
This pilot study will investigate the relative bioavailability of new fixed dose 
combinations (FDC) of ambrisentan and tadalafil, compared to the two monotherapies 
taken concurrently in healthy subjects
Section 2.1 Study Rationale
CHANGE FROM
This study is designed to select one, or more, FDCs of ambrisentan and tadalafil for 
further development and to provide pharmacokinetic data to enable a pivotal 
bioequivalence study (BE). Dependent on formulation work, the study will allow up to 8 
new FDCs to be compared with the reference of ambrisentan and tadalafil monotherapies 
taken concurrently. The study would also allow for up to 2 of the new formulations, that 
may be taken in to a BE study, to be tested for any effect on pharmacokinetics of the 
FDC in both fed and fasted state.
The formulation(s) to be taken forward into the BE study will be primarily chosen on the 
test/reference ratio for both AUC and Cmax for both components ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, between subject variability, 
tablet size, cost, ease of manufacture and stability would be considered.
 
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CHANGE TO
This study is designed initially to compare the relative bioavailability of a number of
fixed dose combinations (FDCs) of ambrisentan and tadalafil (Part 1 and 2) and 
consequently the bioequivalence of the FDC of different dose strengths (Part 3A and 
3B). Dependent on formulation work, the study will allow up to 9 new FDCs to be 
compared with the reference of ambrisentan and tadalafil monotherapies taken 
concurrently
The formulation(s) to be taken forward in Part 3A  will be primarily chosen based on the 
test/reference ratio for both AUC and Cmax for both components. Ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, tablet size, cost, ease of 
manufacture and stability would be considered. 
Part 3 will include evaluation of bioequivalence for 3 dose strengths of the FDC; 
10/40mg, 5/40mg and 5/20mg of ambrisentan and tadalafil.
Section 2.1 Brief Background
CHANGE FROM
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris EMA SmPC, 2015 Section 5.1). Efficacy has been 
shown in idiopathic PAH (IPAH) and in PAH associated with connective tissue 
disease.(Volibris, EMA SmPC,2015). Ambrisentan is an oral, once daily, propanoic acid-
based, ETA-selective Endothelin receptor antagonist (ERA) which targets the 
phospholipase-C-dependent endothelin pathway and which is known to play an essential 
role in mammalian cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (EMEA, 2010;Schuirmann, 
1987;Adcirca, 2015).Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) 
 
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inhibitor which targets the NO pathway. Through inhibition of PDE-5, tadalafil increases 
cytoplasmic cGMP concentrations in the smooth muscle cells and enhances NO-mediated 
vasodilatation of the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy subjects was performed to detect 
any significant PK interactions between tadalafil and ambrisentan when co-administered 
(Spence, 2009). From this study, it was concluded that there is no clinically significant 
PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when combined. 
Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
administered. . There were no SAE’s in the study. Three subjects withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one subject because of myalgia (severe), 
muscle fatigue and dizziness on tadalafil alone and one because of headache (severe) on 
the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild and is a 
listed event for ambrisentan. There were a total of 7 subjects with mild tachycardia. There 
were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. There were 3 events on 
the first day of tadalafil 40 mg given 3 days after ambrisentan 10 mg. There was one 
event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of ambrisentan 10 mg. 
Taken together these data suggest that in healthy subjects a mild tachycardia may result 
from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, EMA SmPC, 
2015;EMEA, 2010;Schuirmann, 1987;Adcirca, 2015).
The AMBITION clinical study (Galiè, 2015a and AMB112565 CSR 13Nov2014), which 
evaluated the time to first clinical failure event, a composite endpoint, shows a robust 
clinical benefit (50% hazard reduction) for PAH patients initiated on a combination of 
ambrisentan and tadalafil when compared to PAH patients initiated on either medication 
as monotherapy.  The safety profile of the combination arm was consistent with the 
known safety data of the individual study drugs and no safety signals specific to 
combination treatment were identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
 
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The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 
CHMP/EWP/240/95) (2008).
CHANGE TO
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris, EMA SmPC, 2015 Section 5.1). Efficacy has been 
shown in idiopathic PAH (IPAH) and in PAH associated with connective tissue disease
(Volibris, EMA SmPC, 2015). Ambrisentan is an oral, once daily, propanoic acid-based, 
ETA-selective Endothelin receptor antagonist (ERA) which targets the phospholipase-C-
dependent endothelin pathway and which is known to play an essential role in 
mammalian cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (EMEA, 2010;Schuirmann, 
1987;Adcirca, 2015). Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) 
inhibitor which targets the NO pathway. Through inhibition of PDE-5, tadalafil increases 
cytoplasmic cGMP concentrations in the smooth muscle cells and enhances NO-mediated 
vasodilatation of the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
 
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Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy participants was performed to 
detect any significant PK interactions between tadalafil and ambrisentan when co-
administered (Spence, 2009). From this study, it was concluded that there is no clinically 
significant PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when 
combined. Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
administered.  There were no SAE’s in the study. Three participants withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one participant because of myalgia 
(severe), muscle fatigue and dizziness on tadalafil alone and one because of headache 
(severe) on the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild 
and is a listed event for ambrisentan. There were a total of 7 participants with mild 
tachycardia. There were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. 
There were 3 events on the first day of tadalafil 40 mg given 3 days after ambrisentan 10 
mg. There was one event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of 
ambrisentan 10 mg. Taken together these data suggest that in healthy subjects a mild 
tachycardia may result from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, EMA SmPC, 
2015;EMEA, 2010;Schuirmann, 1987;Adcirca, 2015)The AMBITION clinical study 
(Galiè, 2015a and GSK document number 2014N193963_00), which evaluated the time 
to first clinical failure event, a composite endpoint, shows a robust clinical benefit (50% 
hazard reduction) for PAH patients initiated on a combination of ambrisentan and 
tadalafil when compared to PAH patients initiated on either medication as monotherapy.  
The safety profile of the combination arm was consistent with the known safety data of 
the individual study drugs and no safety signals specific to combination treatment were 
identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
 
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and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 2008).
Section 4.1 Overall Design
CHANGE FROM 
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects. See Figure 1 for study schematic.
Figure 2
Study Schematic
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
 
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There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose. 
The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast.
Subjects who had taken part in the study part preceding Part 3, and which provided the 
successful formulations for a BE study could also be invited to take part in this study 
part. However, a subject’s inclusion in more than one study part would be dependent on 
the subject not exceeding the maximum blood draw volume (500ml) for the study.
 
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CHANGE TO
This is a single centre, Phase 1, single dose, randomised, open label, crossover study in 
healthy volunteers with 3 study parts; Part 1 will include a 5 way cross-over, and 
Part 2 and 3 (A&B) will each include a 4 way cross-over. See Figure 1 for study 
schematic.
Figure 1
Study Schematic
Screening
≤ 31 days
Follow 
Up
Screening 
≤ 31 days from
Subjects 
Randomised to  
4 dose sessions 
(3 granulation 
formulations + 
reference)
Follow 
Up
Subjects 
randomised to 4 
dose sessions 
(FDC Fed & 
fasted + 
reference (fed 
&fasted))
Screening 
≤ 31 days
Follow 
Up
~4 Weeks
~4 Weeks
~4 Weeks
PART 1 (BA) n=20
PART 2 (BA) n=20
PART 3A (BE 10/40) n=33
Subjects 
Randomised to 
up to 5 dose 
sessions (4 FDC 
formulations 
and the 
reference)
Screening 
≤ 31 days
Subjects 
Randomised to 
4 dose sessions    
(2 FDC 
formulations 
(fasted) + 2 
reference fasted)
Follow 
Up
PART 3B (BE 5/40 & 5/20)
n=33
~4 Weeks
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in study Part 1 and 2. In 
study Part 3 there will be a minimum of 10 days between each dose.  A Follow-Up 
Visit should be completed within 7-14 days of completion of a subject’s last dose. 
The utility of each of the 3 study parts is as follows:
 
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Part 1
This study part, will have 5 dose sessions and will be used  to characterise the relative 
bioavailability, safety and tolerability of   four formulations of the fixed dose 
combination (ambrisentan 10 mg + tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
Part 2
This study part will have 4 dose sessions and will evaluate the bioavailability, safety and 
tolerability of  3 different granulation sizes for a single FDC (ambrisentan 10 mg + 
tadalafil 40 mg) compared to the reference of the 2 monotherapy components taken 
concurrently (ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted 
state.
Part 3A
Part 3A of the study is set to establish bioequivalence between the candidate FDC 
from Part 2. This study part will have 4 dose sessions and will assess the 
bioequivalence, in both the fed and fasted state, of the FDC against the reference  
ambrisentan 10 mg + tadalafil 40mg monotherapies in the fed and fasted state. The 
fed arms of this part will have a standard high fat breakfast (EMEA, 2010).
Part 3B
Part 3B of the study is set to establish bioequivalence between the Fixed Dose 
Combination of an additional two dose strengths (ambrisentan 5 mg + tadalafil 
40mg and ambrisentan 5 mg + tadalafil 20mg) . This study part will have 4 dose 
sessions and will assess the bioequivalence of the two FDC dose strengths against the 
reference ambrisentan 5 mg + tadalafil 40mg monotherapies and ambrisentan 5 mg 
+ tadalafil 20mg monotherapies in the fasted state.
Section 4.2 Treatment and Duration
CHANGE FROM
The treatments for each study part of the study are listed in Table 1 All treatments are 
single dose. Subjects will be randomised to order of treatments in the parts of the study 
they are included in.
The study has 3 parts and ongoing analyse of pharmacokinetic data will be used to enable 
the formulations produced and tested in subsequent parts.
Part 1
Part 1 of the study will be utilised to look at up to 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
 
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following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2. 
If no successful formulations are identified from these criteria then the pharmacokinetic 
data would be used to enable further work to produce new FDC formulations for Part 2
Following completion of Part 1 there will be a pause prior to Part 2, so that up to a further 
4 FDC formulations could be produced and data included and approved in any required 
update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for up to 4 FDC formulations. These could be 
either successful formulation identified in Part 1 reformulated with the final API and/or 
new formulations using the final API. Pharmacokinetic data from Part 2 of the study will 
be analysed following completion of each treatment arm by subjects. This ongoing 
review of the pharmacokinetic data will enable rapid ongoing assessment of each FDC 
formulation and will be used to define any successful FDC formulation to be taken into 
Part 3 of this study and a pivotal BE study. If only two formulations are evaluated in Part 
2 then the food effect may be added to Part 2 and Part 3 will not be required. Success will 
be defined with the same criteria as those in Part 1.
Part 3
Part 3 of the study will provide data for up to 2 successful formulations to be progressed 
to a pivotal BE study, in both the fed and fasted state.
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil FDC5 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil FDC6 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil FDC7 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC4 (10mg/40mg)
ambrisentan and tadalafil FDC8 
(10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. Subjects may be included in Part 1, Part 2 or Part 3 of the study. However, if Part 2 
of the study identifies any successful formulations to take into Part 3 then subjects from 
Part 2 may be invited to join Part 3.
The minimum and maximum duration for a subject on this study is described in Figure 2. 
This table assumes all 5 dose sessions are used for Part 1 and Part 2 and either 2 or 4 dose 
sessions are used in Part 3. However, this is an approximation as fewer sessions may be 
required in Part 2 and/or the washout between sessions of 7 days in each part may be 
longer, due to logistical reasons.
 
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Table 2
Study Duration
Study Options
Screen
Study 
Part 
1or 2
Pause
Study 
part 3
Follow-
Up Visit
Total 
Duration 
on Study
(1-31 
days)
(31 
days)
(7-28 
days)
(10-24 
days)
(7-14 
days)
Subjects 
included 
in only 
Part 1 or 
2
Min
1
31
7
39
Max
31
31
14
76
Subjects 
included 
in Part 
2&3
Min
1
31
7
10
7
56
Max
31
31
28
24
14
128
Subjects 
included 
in only 
Part 3
Min
1
10
7
18
Max
31
24
14
69
A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
CHANGE TO
The treatments for each study part of the study are listed in Table 1. All treatments are 
single dose. Subjects will be randomised to the order of treatments in the parts of the 
study they are included in.
The study has 3 parts and ongoing analysis of pharmacokinetic data will be used to 
determine the formulations produced and tested in subsequent parts.
Part 1
Part 1 of the study will be utilised to evaluate 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
 
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FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2
and 3. 
Following completion of Part 1 there will be a pause prior to Part 2, so that different 
granulation forms of a single FDC can be produced and data included and approved in 
any required update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for 3 granulation size forms of a single FDC 
formulation selected from Part 1. These will be based on a single successful 
formulation identified in Part 1 reformulated with the final active pharmaceutical 
ingredient (API). Success will be defined with the same criteria as those in Part 1.
Part 3 (A and B)
The BE part of the study will investigate the bioequivalence of the FDC ambrisentan 
10 mg + tadalafil 40 mg in part 3A and the bioequivalence of the FDC ambrisentan 
5 mg + tadalafil 40 mg and ambrisentan 5 mg + tadalafil 20 mg in Part 3B.  
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted) 
Part 3A (fed and fasted)
Part 3B (fasted)
ambrisentan and 
tadalafil FDC1 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 1 
(10mg/40mg)
ambrisentan and 
tadalafil FDC
(10mg/40mg), fed
Ambrisentan 
and tadalafil 
FDC 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC2 
(10mg/40mg)
ambrisentan and tadalafil 
FDCgranulation 2
(10mg/40mg)
ambrisentan and 
tadalafil FDC
(10mg/40mg) fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/40mg), 
fasted
ambrisentan and 
tadalafil FDC3 
(10mg/40mg)
ambrisentan and tadalafil 
FDC granulation 3
(10mg/40mg)
ambrisentan and 
tadalafil monotherapies 
taken concurrently
(10mg/40mg), fed
ambrisentan 
and tadalafil 
FDC 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil FDC4 
(10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently
(10mg/40mg)
ambrisentan and 
tadalafil monotherapies 
taken concurrently
(10mg/40mg), fasted
ambrisentan 
and tadalafil 
monotherapies 
taken 
concurrently 
(5mg/20mg), 
fasted
ambrisentan and 
tadalafil 
monotherapies taken 
concurrently 
(10mg/40mg)
The duration for a subject on this study is described in Figure 2. This figure assumes all 5 
dose sessions are used for Part 1. However, this is an approximation as minimum of 7 
days is expected between doses in Parts 1 and 2 and a minimum of 10 days in Part 3.
 
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Figure 2
Study Duration
Part 1 can last up to 73 days.
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
Dose 
Period 5 
start 
7 
7 
7 
7 
7 to 
14 
Follow-up
 
 
 
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Part 2  will last approximately 9 weeks
Part 3A and B will last approximately 11 weeks
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start 
Dose 
Period 4 
start
10 
days
10  
days
10 
days
7 to 14  
days
Follow-up
Screening
Up to 31 days
Dose 
Period 1 
start 
Dose 
Period 2 
start 
Dose 
Period 3 
start
Dose 
Period 4 
start
7 days
7 days
7 days
7 to 14  
days
Follow-up
 
 
 
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A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
Section 4.3 Type and Number of Subjects
CHANGE FROM
A approximately of 20 healthy adult subjects will be randomized, to each study part, such 
that at least 16 evaluable subjects complete each part of the study.
The study is not powered. However, the number of subjects will be sufficient for the 
objectives of this study and this is detailed further in Section 9.2 Sample Size 
Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
CHANGE TO
Approximately 20 healthy adult subjects will be randomized to study Part 1 and an 
additional 20 healthy adult subjects in Part 2, such that at least 16 evaluable subjects 
complete each part of the study.  Approximately 33 healthy subjects will be 
randomized to study Part 3A, such that at least 26 evaluable subjects complete part 
3A of the study. An additional 33 healthy subjects will be randomised to Part 3B, 
such that at least 26 evaluable subjects complete part 3B of the study.
The estimation of the number of subjects for each study part is detailed further in 
Section 9.2 Sample Size Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
Section 4.4 Design Justification
CHANGE FROM
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations to take into a pivotal BE study and to also provide pharmacokinetic data to 
enable the pivotal BE study to be powered sufficiently. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
 
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The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, or is there any need to blind study treatment. The Inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study is in three parts to enable PK data from Part 1 to enable formulation 
development for Part 2. Part 3 providing an opportunity to assess food effect in any FDC 
formulations suitable for development and inclusion in the pivotal bioequivalence study. 
The interdependency of the 3 study parts is described in detail in Section 4.2. 
CHANGE TO
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, nor is there any need to blind study treatment. The inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study contains three parts: Parts 1 and 2 will provide PK data for different 
candidate FDC formulations and will enable the assessment of bioequivalence of the 
FDC versus the combination of the individual reference products in Part 3. The 
interdependency of the 3 study parts is described in detail in Section 4.2. 
Section 4.5 Dose Justification
CHANGE FROM
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of, 40mg for tadalafil and 10mg for ambrisentan are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
be tolerated in healthy subjects.
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimum washout of 7 days, 
between doses, is also sufficient for clearance of previous dose.
 
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CHANGE TO
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of 40mg for tadalafil and 10mg for ambrisentan, are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
be tolerated in healthy subjects. Within Part 3 of the study all dosage forms of the 
FDC that are proposed for marketing authorisation will be tested for bioequivalence 
versus the appropriate doses of reference products.  These doses are required in 
clinical practice to allow safe up- and down-titration of the FDC in patients with 
PAH.  
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimumof 10 days, between 
doses, is also sufficient for clearance of the previous dose during the assessmet of 
bioequivalence.
Section 4.6.2 Benefit Assessment
CHANGE FROM
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation for a 
fixed dose combination of ambrisentan plus tadalafil to use as first line therapy in PAH 
patients.
CHANGE TO
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation and to 
demonstrate bioequivalence versus reference products for a fixed dose combination of 
ambrisentan plus tadalafil to use as first line therapy in PAH patients.
Section 4.6.3 Overall Benefit: Risk Conclusion
CHANGE FROM
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 3. So, the 
risk: benefit is appropriate for this study and to enable development of this investigational 
product. The overall benefit: risk therefore remains positive.
 
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CHANGE TO
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 3. In 
addition review of safety information from Part 1 of the study is consistent with this 
position. So, the benefit:risk is appropriate for this study and to enable development of 
this investigational product. The overall benefit: risk therefore remains positive.
Section 6.1 Investigational Product and Other Study Treatment
CHANGE FROM
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Part 1 of the study are shown in Table 4, this will be 
amended once treatments for Part 2 and Part 3 are confirmed.
 
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Table 4
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS*
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS*
*
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 95 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 233 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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CHANGE TO
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Parts 1, 2 and 3 of the study are shown in Table 4, 
Table 5 and Table 6.
 
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Table 4
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 90 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.45
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Address of 
reference 
manufacturer
NA
NA
NA
NA
Glaxo Operations UK 
Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 5
Study Treatment for Part 2
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 granulation 
1 (10mg/40mg)
FDC-G2 granulation 2 
(10mg/40mg)
FDC-G3 granulation 
3 (10mg/40mg)
Each tablet contains 10 
mg of ambrisentan, 
approximately 90 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg of 
lecithin (soya) (E322) and 
approximately 0.45 mg of 
Allura red AC Aluminium 
Lake 
Each film-coated 
tablet contains 20 mg 
tadalafil, 245 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage level(s):
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 10 
mg of ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration 
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC
FDC
FDC
Reference
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations UK Ltd 
(trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 8DT
United Kingdom
Lilly S.A 
Avda.  de la Industria 
30
28108 Alcobendas
Madrid
Spain
 
 
 
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Table 6
Study Treatment for Part 3
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-
Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil -
Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Formulation 
description
FDC-G1 or 2 or3
(10mg/40mg)
FDC  (5mg/40mg)
FDC  (5mg/20mg)
Each tablet contains 
10 mg of 
ambrisentan, 
approximately 90 mg 
of lactose (as 
monohydrate), 
approximately 0.25 
mg of lecithin (soya) 
(E322) and 
approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each tablet 
contains 5 mg of 
ambrisentan, 
approximately 95 
mg of lactose (as 
monohydrate), 
approximately 
0.25 mg of 
lecithin (soya) 
(E322) and 
approximately 
0.11 mg of Allura 
Red AC 
Aluminium Lake
Each film-coated 
tablet contains 20 
mg tadalafil, 245 
mg lactose (as 
monohydrate).
Dosage form:
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 5 mg 
of ambrisentan.
Each tablet contains 
20 mg tadalafil and 5 
mg of ambrisentan.
Each tablet contains 
10 mg of 
ambrisentan.
Each tablet 
contains 5 mg of 
ambrisentan.
Each tablet 
contains 20 mg 
tadalafil.
Route of 
Administration 
Oral
Oral
Oral
Oral
Oral
Oral
 
 
 
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Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK1325760 
(ambrisentan-
Volibris)
GSK1325760 
(ambrisentan-
Volibris)
GF196960 
(tadalafil -
Adcirca)
Treatment 
FDC
FDC
FDC
Reference
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Address of 
reference 
manufacturer
NA
NA
NA
Glaxo Operations 
UK Ltd (trading as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Glaxo Operations 
UK Ltd (trading 
as 
GlaxoWellcome 
Operations)
Harmire Road
Barnard Castle
Co. Durham DL12 
8DT
United Kingdom
Lilly S.A 
Avda.  de la 
Industria 30
28108 
Alcobendas
Madrid
Spain
 
 
 
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6.1.1 Retention Samples
GSK will send five times (5x) the drug product necessary for pivotal bioequivalency 
evaluation in Part 3 of the study for analytical testing for all tablet strengths used in 
the study. The retain samples are in addition to the supply of drug product 
sufficient to complete the study. Retention samples will be sent to the clinical CRO 
in the same bulk container. To ensure that reserve samples are representative of the 
same batches provided for the clinical study the clinical site will randomly select the 
supplies for the study from the supply received. The clinical site must retain enough 
reserve samples to permit FDA to perform five times all of the release tests required 
in the application.
Section 6.2 Treatment Assignment
CHANGE FROM
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software.
The treatments are denoted as F1 to F4 for Part 1, F5 to F8 for Part 2 and R for reference 
treatment for both parts; the selected formulations for Part 3 are denoted as X and Y.  The 
treatment key for Parts 1, 2 and 3 are described in Table 6
Table 6 
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 2
F5
ambrisentan and tadalafil FDC5 (10mg/40mg)
F6
ambrisentan and tadalafil FDC6 (10mg/40mg)
F7
ambrisentan and tadalafil FDC7 (10mg/40mg)
F8
ambrisentan and tadalafil FDC8 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3
 
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Treatment
Description
X1
ambrisentan and tadalafil FDCX1 (10mg/40mg), fed
X2
ambrisentan and tadalafil FDCX1 (10mg/40mg) fasted
Y1
ambrisentan and tadalafil FDCX2 (10mg/40mg), fed
Y2
ambrisentan and tadalafil FDCX2 (10mg/40mg), fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 7. Not all possible sequences are 
included here. Such as, if only one formulation taken into Part 2 or 3, or Part 2 is used for 
only one or two FDC formulations, fed and fasted. Additional treatment sequences will 
be created based on the Latin Squares for Williams Designs, as required
Table 7 
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1 F2 R F3 F4
F2 F3 F1 F4 R
F3 F4 F2 R F1
F4 R F3 F1 F2
R F1 F4 F2 F3
F4 F3 R F2 F1
R F4 F1 F3 F2
F1 R F2 F4 F3
F2 F1 F3 R F4
F3 F2 F4 F1 R
1:1:1:1:1:1:1:1:1:1
Part 2
5
F5 F6 R F7 F8
F6 F7 F5 F8 R
F7 F8 F6 R F5
F8 R F7 F5 F6
R F5 F8 F6 F7
F8 F7 R F6 F5
R F8 F5 F7 F6
F5 R F6 F8 F7
F6 F5 F7 R F8
F7 F6 F8 F5 R
1:1:1:1:1:1:1:1:1:1
4
F5 F6 R F7
F6 F7 F5 R
F7 R F6 F5
R F5 F7 F6
1:1:1:1
3
F5 F6 R
R F6 F5
F6 R F5
F1 R F6
R F5 F6
F6 F5 R
3:3:3:3:3:3 + 2 random
2
F5 R 
1:1
 
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Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
R F5
Part 3
4
X1 X2 Y2 Y1
X2 Y1 X1 Y2
Y1 Y2  X2 X1
Y2  X1 Y1 X2 
1:1:1:1
Part 2 and 
Part 3 
merged 
together
5 (Two FDCs selected from 
Part 1)
X1 X2 R Y1 Y2
X2 Y1 X1 Y2 R
Y1 Y2 X2 R X1
Y2 R Y1 X1 X2
R X1 Y2 X2 Y1
Y2 Y1 R X2 X1
R Y2 X1 Y1 X2
X1 R X2 Y2 Y1
X2 X1 Y1 R Y2
Y1 X2 Y2 X1 R
1:1:1:1:1:1:1:1:1:1
3 (One FDC selected from 
Part 1)
X1 X2 R
R X2 X1
X2 R X1
F1 R X2
R X1 X2
X2 X1 R
3:3:3:3:3:3 + 2 random
CHANGE TO
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software. The treatments are 
denoted as F1 to F4 for Part 1, F5 to F8 for Part 2 and R for reference treatment for both 
parts; the selected formulations for Part 3 are denoted as X and Y
The treatment key for Parts 1, 2 and 3 are described in Table 6.
Table 6
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
 
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Treatment
Description
Part 2
FG1
ambrisentan and tadalafil FDC-G1 (10mg/40mg) granulation size 1
FG2
ambrisentan and tadalafil FDC-G2 (10mg/40mg) granulation size 2
FG3
ambrisentan and tadalafil FDC-G3 (10mg/40mg) granulation size 3
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3A
X1
ambrisentan and tadalafil FDC-G1 or 2 or 3  (10mg/40mg) fed
X2
ambrisentan and tadalafil FDC G1 or 2 or 3 (10mg/40mg) fasted
R1
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fed
R2
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg), fasted
Part 3B
Y1
ambrisentan and tadalafil FDC (5mg/40mg), fasted
Y2
ambrisentan and tadalafil FDC (5mg/20mg), fasted
R3
ambrisentan and tadalafil monotherapies taken concurrently 
(5mg/40mg), fasted
R4
ambrisentan and tadalafil monotherapies taken concurrently 
(5mg/20mg), fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 7. Not all possible sequences are 
included here. Such as, if only one formulation taken into Part 2 or 3, or Part 2 is used for 
only one or two FDC formulations, fed and fasted Additional treatment sequences may be 
created based using the Latin Squares for Williams Designs if needed.
Table 7
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1/ F2/ R/ F3/ F4
F2/ F3/ F1/ F4/ R
F3/ F4/ F2/ R/ F1
F4/ R/ F3/ F1/ F2
R/ F1/ F4/ F2/ F3
F4/ F3/ R/ F2/ F1
R/ F4/ F1/ F3/ F2
F1/ R/ F2/ F4/ F3
F2/ F1/ F3/ R/ F4
F3/ F2/ F4/ F1/ R
1:1:1:1:1:1:1:1:1:1
 
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Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 2
4
FG1/ FG2/ R/ FG3
FG2/ FG3/ FG1/ R
FG3/ R/ FG2/ FG1
R/ FG1/ FG3/ FG2
1:1:1:1
Part 3A
4
X1/ R1/ R2/ X2
R1/ X2/ X1/ R2
X2/ R2/ R1/ X1
R2/ X1/ X2/ R1
1:1:1:1
Part 3B
4
Y1/ R3/ R4/ Y2
R3/ Y2/ Y1/ R4
Y2/ R4/ R3/ Y1
R4/ Y1/ Y2/ R3
1:1:1:1
Section 9.1 Hypotheses
CHANGE FROM
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and if used, Study part 2, (ii) estimate the bioavailability of the 
formulation(s) of the FDC formulations, taken in to Part 3, in the fed state relative to the 
fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
CHANGE TO
No formal hypothesis will be tested for study Part 1 and Part 2. An estimation 
approach  will be used to (i) estimate the bioavailability of the FDC formulations relative 
to the monotherapies taken concurrently, (ii) for each primary pharmacokinetic 
endpoint, point estimates and corresponding 90% confidence intervals will be 
constructed for the ratio of the geometric mean of the test treatment (FDC 
formulations) to the geometric mean of the reference treatment (co-administration 
of ambrisentan and tadalafil), (test)/ (reference).  The objective of Part 2 is to 
assess whether differences in granulation size impact the pharmacokinetics of 
ambrisentan and tadalafil; the estimation approach for bioavailability is therefore 
applicable to Part 2.
Part 3A of the study is designed to test the bioequivalence of a FDC of ambrisentan 
10 mg + tadalafil 40 mg (test) relative to reference monotherapies of  ambrisentan 
10 mg & tadalafil 40 mg taken concurrently (reference) in healthy subjects in both 
the fed and fasted states. The null hypothesis is that the true ratio of the geometric 
 
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mean of the test treatment to the geometric mean of the reference treatment, 
(test)/(reference), for each primary PK endpoint, is either less than 0.80 or 
greater than 1.25.  The alternate hypothesis is that the true ratio of the test 
treatment geometric mean to the reference treatment geometric mean is at least 0.80 
and no greater than 1.25.  Symbolically, this is expressed as follows:
H(0): (test)/(reference) < 0.80 or (test)/(reference) > 1.25,
i.e., treatments are not bioequivalent.
versus
H(1) : 0.80 ( test)/(reference) 1.25,
i.e. , treatments are bioequivalent.
For each PK parameter designated as a primary endpoint, a two one-sided t-test 
(TOST) procedure (Schuirmann, 1987) with =0.05 for each one-sided test will be 
used to test this set of hypotheses.  This is equivalent to requiring that a 90% 
interval for the true ratio of test to reference geometric means falls entirely within 
the range of 0.80 to 1.25.
Part 3B of the study is designed similar to Part 3A and will test (1) the 
bioequivalence of a FDC of ambrisentan 5 mg + tadalafil 40 mg (test) relative to 
reference monotherapies of ambrisentan 5 mg & tadalafil 40 mg taken concurrently 
(reference) and (2) the bioequivalence of a FDC of ambrisentan 5 mg + tadalafil 20 
mg (test) relative to reference monotherapies of ambrisentan 5 mg & tadalafil 20 mg 
taken concurrently (reference), in healthy subjects in the fasted states. Therefore, 
the analysis approach for bioequivalence for Part 3A is applicable to Part 3B.
Section 9.2 Sample Size Considerations
CHANGE FROM:
No formal sample size calculation has been performed. Sample sizes chosen for each 
study part in this study is considered sufficient for exploratory analysis based on PK 
variability data from previous studies
CHANGE TO
No formal sample size calculation has been performed. Sample sizes chosen for each 
study part in this study is considered sufficient for exploratory analysis based on PK 
variability data from previous studies
Section 9.2.1 Sample Size Assumptions
 
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CHANGE FROM
The sample size assumptions are based on previously reported estimates of within subject 
CV for AUC(0-∞) and Cmax for ambrisentan (GS-US-300-0112, 2008) and tadalafil 
(Forgue, 2005). Table 9 summarizes the estimates of within subject CV for the primary 
endpoints AUC (0-∞) and Cmax.
Table9 
Estimates of within subject CV for the primary end points AUC (0-∞)
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
CHANGE TO
For Part 1 and Part 2 of the study, the sample size assumptions are based on 
previously reported estimates of within subject CV for AUC(0-∞) and Cmax for 
ambrisentan (GS-US-300-0112, 2008) and tadalafil (Forgue, 2005). Table 9 summarizes 
the estimates of within subject CV for the primary endpoints AUC (0-∞) and Cmax.
Table 9 
Estimates of within subject CV for the primary end points AUC (0-∞) 
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
For Part 3A of the study, Table 10 summarized the observed estimates of between 
and within subject CV and observed ratio of geometric means for the primary 
endpoints AUC(0-t) and Cmax from Part 1 of the study.
 
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Table 10
Estimates of within subject CV for the primary end points AUC 
(0-∞) and Cmax for ambrisentan and tadalafil from part 1 of the study
CVw: within subject CV
ambrisentan 
tadalafil 
Cmax
22.42%
14.53%
AUC (0-∞)
8.12%
10.32%
The largest of the within subject CV estimates 22.42% translates to a standard 
deviation (SD) of 0.221 on the natural log scale.  Based on this SD, a sample size of 
23 statistically evaluable subjects will have 90% power to demonstrate 
bioequivalence. This calculation assumes:

a true ratio of 1,

the within-subject variability from the current study will not be larger than 
that used in the sample size calculations,

data are log-normally distributed, and each t-test is made at the 5% level.
For Part 3B of the study, same assumptions are adopted as Part 3A. Thus, a sample 
size of 23 statistically evaluable subjects will have 90% power to demonstrate 
bioequivalence.  However, if the within subject CVs obtained after Part 2 are 
significantly higher than the ones in Part 1 (shown in Table 10), a sample size re-
esimation may be conducted to adjust the sample size for Part 3A and 3B to ensure 
the study maintain enough power to demonstrate bioequivalence.
Section 9.2.2 Sample Size Sensitivity
CHANGE FROM
Assuming the within subject CV differs from 22% and the number of statistically 
evaluable subjects is lower than 20, then Table 11 shows the precision (the half width of 
the 90% confidence interval) for a range of evaluable subjects.
 
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Table 11
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
CHANGE TO
For Part 1 and Part 2, assuming the within subject CV differs from 22% and the 
number of statistically evaluable subjects is lower than 20, then Table 11 shows the 
precision (the half width of the 90% confidence interval) for a range of evaluable 
subjects.
Table 11
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
For Part 3, based on the observed data from previous studies, the ratio of geometric 
mean for ambrisentan Cmax is unlikely to be 1. The effects on the power of 
declaring bioequivalence in the face of a shift in the expected ratio of the geometric 
means were examined.
 
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If, under all other assumptions outlined above, the actual ratio of geometric means 
for ambrisentan Cmax is 0.97, then 26 evaluable subjects are needed to have 90% 
power to conclude bioequivalence.
With 26 evaluable subjects, the current study design has at least 90% power to 
conclude bioequivalance (with both Cmax and AUC for ambrisentan and tadalafil).
Section 9.2.3 Sample Size Re-estimation or Adjustment
CHANGE FROM
No sample size re-estimation will be performed.
CHANGE TO
No sample size re-estimation will be performed.  However, if the estimation of the 
within subject CV from Part 2 of the study is much larger than the within subject 
CV from Part 1 of the study, the sample size for Part3 may be re-evaluated.
Section 9.3 Data Analysis Considerations
CHANGE FROM
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
sampling times recorded during the study.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-
life (t1/2).  
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model.
CHANGE TO
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
 
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sampling times recorded during the study although supplementary analayis will be 
available based on the nominal times.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-life 
(t1/2).  
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model
9.3.1
Analysis Populations
9.3.1.1.
Safety Population
All subjects enrolled into the study who have received at least one dose of investigational 
product will be included in the Safety Population.
9.3.1.2.
Pharmacokinetic Concentration Population
The PK Concentration Population will include all subjects for whom at least one PK 
sample was obtained and analyzed.
9.3.1.3.
Pharmacokinetic Parameter Population
For each PK parameter, the PK Parameter Population will include all subjects who 
provide PK parameter data.
Section 9.3.2 Interim Analysis
CHANGE FROM
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in order to direct development of fixed dose formulations for subsequent 
study parts and the planned BE study.
CHANGE TO
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in Part 1 and Part 2 of the study in order to direct development of the 
FDC for later parts of the study.  Headline results based on statistical analysis using 
preliminary pharmacokinetic data with nominal time may be produced when 80% 
and 100% subjects complete Part 1 and Part 2 of the study to assist development of 
 
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FDCs.  Treatment and period information from the crossover design may be used in 
the analysis; sequence information may also be included depending on availability.
Section 9.4.1 Pharmacokinetic Analyses
CHANGE FROM
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D. 
Following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using a mixed effects model with fixed effect 
terms for period and treatment. Subject will be treated as a random effect in the model. 
Point estimates and their associated 90% confidence intervals will be constructed for the 
differences, F1 – R, F2 – R, F3 – R, F4 – R for Part 1 and similarly for Part 2 (as needed); 
and X1-X2, Y1-Y2 for Part 3. The point estimates and their associated 90% confidence 
intervals will then be back-transformed to provide point estimates and 90% confidence 
intervals for the ratios, F1:R, F2-R:F3-R, F4-R; and X1:X2, Y1:Y2.
Tmax of FDC formulations will be analyzed with the non-parametric Wilcoxon Matched 
Pairs Method to compute point estimate and associated 90% confidence intervals for the 
median difference, F1 R, F2 R, F3 R, F4 R and X1:X2, Y1:Y2.
CHANGE TO
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D.
Data will be listed and summarized according to GlaxoSmithKline reporting 
standards, where applicable.  Listings will be sorted by subject, period, noting 
treatment.  Summaries will be presented by treatment.  Unless stated otherwise, 
descriptive summaries will include n, mean, standard deviation (SD), coefficient of 
variation (CV), median, minimum, and maximum for continuous variables, n and 
percent for categorical variables and geometric mean, 95% confidence interval (CI), 
and the between-subject CV (CVb) based on geometric mean for the log-
transformed PK parameters.
Version 9.3 (or higher) of the SAS system will be used for statistical analysis of the 
data as well as to generate tables, figures, and listings.
 
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Any deviation(s) from the original analyses planned in the protocol will be reported 
in the Reporting and Analysis Plan (RAP) and/or in the Clinical Pharmacology 
Study Report.
The PK parameters, AUC(0-), AUC(0-t), Cmax, and t½ will be transformed using 
natural logarithms. Missing PK parameters will not be imputed.
All data from withdrawn subjects will be listed.
No adjustment for multiple tests or comparisons is planned.
Actual visit day will be used for safety assessments. For the assessment of 
bioequivalence, both actual time of sampling, and nominal (planned) sampling time, 
will be used to estimate PK parameters at the end of the study.  PK parameters 
estimation based on nominal sampling time will be used for generating headline 
results.Following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using a mixed effects model with fixed effect 
terms for period, sequence and treatment, and subject as random effect term (i.e. 
ANOVA method using period, sequence and treatment as fixed effect, and subject as 
random effect) Point estimates and their associated 90% confidence intervals will be
calculated for the differences: F1 - R, F2 - R, F3 -R, F4 - R for Part 1; FG1-R, FG2-R, 
FG3-R for Part 2; X1-R1, X2-R2 for Part 3A; and Y-R3 for Part 3B. The point 
estimates and their associated 90% confidence intervals will then be back-transformed to 
provide point estimates and 90% confidence intervals for the ratios, F1:R, F2:R, F3:R, 
F4:R; for Part 1; FG1:R, FG2:R, FG3:R for Part 2; X1:R1,X2:R2 for Part3A; and 
Y:R3 for Part 3B.
Section 11 REFERENCES
ADDITIONS
EMEA guidelines on the investigation of bioequivalence. CPMP/EWP/QWP/1401/98 
Rev. 1/ Corr **. 2010
Schuirmann DJ. A comparison of the two one-sided tests procedure and the power 
approach for assessing the equivalence of average bioavailability, J Pharmacokin. 
Biopharm. 1987:15:657–680. 
Section 12.4 Appendix 4 High Fat Meal Content
CHANGE FROM

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
 
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The standard high-fat meal will be the meal suggested by the US FDA in their 2002 draft 
guidance on food-effect bioavailability and bioequivalence studies. Approximately 50% 
of the caloric content of the meal is from fat and the meal is high in calories 
(approximately 1000 calories). Substitutions in this test meal can be made as long as the 
meal provides a similar amount of calories from protein, carbohydrate, and fat and has 
comparable meal volume and viscosity.
US Department of Health and Human Services.  Guidance for Industry. Food-Effect 
Bioavailability and Fed Bioequivalence Studies.  Guidance. Center for Drug Evaluation 
and Research (CDER). Food and Drug Administration; 2002.
CHANGE TO

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
The standard high-fat meal will be the meal suggested by the EMEA guidelines on the 
investigation of bioequivalence 2010. The meal should be a high-fat (approximately
50 percent of total caloric content of the meal) and high-calorie (approximately 800 to
1000 kcal) meal. This test meal should derive approximately 150, 250, and 500-600 
kcal from protein, carbohydrate, and fat, respectively(EMEA, 2010).
 
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TITLE PAGE
Division: Worldwide Development
Information Type: Protocol Amendment
Title:
A Phase 1 study to demonstrate the relative bioavailability of 
fixed dose combinations of ambrisentan and tadalafil in healthy 
subjects
Compound Number:
GSK3380154, GSK1325760 and GF196960
Development Phase:
I
Effective Date:
14-APR-2016
Protocol Amendment Number:  01
Author (s):
 
Revision Chronology
GlaxoSmithKline
Document Number
Date
Version
2015N232335_00
2015-DEC-16
Original
2015N232335_01
2016-APR-14
Amendment No. 1
Clarification of hypotension withdrawal criteria in Section 5.4.3, change to GSK medical 
monitor and minor administrative changes.
Copyright 2016 the GlaxoSmithKline group of companies.  All rights reserved.  
Unauthorised copying or use of this information is prohibited.
 
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MEDICAL MONITOR/SPONSOR INFORMATION PAGE
Medical Monitor/SAE Contact Information: 
Role
Name
Day Time Phone Number and 
email address
After-hours Phone/Cell/
Pager Number
Fax 
Number
Primary 
Medical 
Monitor
Site to 
scan and 
send to 
email
Secondary 
Monitor
Site to 
scan and 
send to 
email
SAE 
contact  
information 
Site to 
scan and 
send to 
email
Sponsor Legal Registered Address: 
GlaxoSmithKline Research & Development Limited
980 Great West Road
Brentford
Middlesex, TW8 9GS
UK
In some countries, the clinical trial sponsor may be the local GlaxoSmithKline Affiliate 
Company (or designee). If applicable, the details of the alternative Sponsor and contact 
person in the territory will be provided to the relevant regulatory authority as part of the 
clinical trial application.   
Regulatory Agency Identifying Number(s): EudraCT number 2015-004140-18
 
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INVESTIGATOR PROTOCOL AGREEMENT PAGE
For protocol number 201964
I confirm agreement to conduct the study in compliance with the protocol, as amended by 
this protocol amendment.
I acknowledge that I am responsible for overall study conduct. I agree to personally 
conduct or supervise the described study.
I agree to ensure that all associates, colleagues and employees assisting in the conduct of 
the study are informed about their obligations. Mechanisms are in place to ensure that site 
staff receives the appropriate information throughout the study. 
Investigator Name:
Investigator Address:
Investigator Phone Number:
Investigator Signature
Date
 
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TABLE OF CONTENTS
PAGE
1.
PROTOCOL SYNOPSIS FOR STUDY 201964........................................................8
2.
INTRODUCTION....................................................................................................11
2.1.
Study Rationale ..........................................................................................11
2.2.
Brief Background ........................................................................................11
3.
OBJECTIVE(S) AND ENDPOINT(S) ......................................................................14
4.
STUDY DESIGN ....................................................................................................14
4.1.
Overall Design ............................................................................................14
4.2.
Treatment and Duration..............................................................................16
4.3.
Type and Number of Subjects.....................................................................19
4.4.
Design Justification.....................................................................................20
4.5.
Dose Justification........................................................................................20
4.6.
Benefit: Risk Assessment ...........................................................................20
4.6.1.
Risk Assessment .........................................................................21
4.6.2.
Benefit Assessment.....................................................................23
4.6.3.
Overall Benefit: Risk Conclusion..................................................23
5.
SELECTION OF STUDY POPULATION AND WITHDRAWAL CRITERIA .............23
5.1.
Inclusion Criteria.........................................................................................24
5.2.
Exclusion Criteria........................................................................................25
5.3.
Screening/Baseline/Run-in Failures............................................................26
5.4.
Withdrawal/Stopping Criteria.......................................................................27
5.4.1.
Liver Chemistry Stopping Criteria ................................................27
5.4.1.1.
Study Treatment Restart or Rechallenge....................28
5.4.2.
QTc Stopping Criteria ..................................................................28
5.4.3.
Hypotension.................................................................................29
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria .........................29
5.5.
Subject and Study Completion....................................................................29
6.
STUDY TREATMENT ............................................................................................29
6.1.
Investigational Product and Other Study Treatment....................................29
6.2.
Medical Devices..........................................................................................31
6.3.
Treatment Assignment................................................................................31
6.4.
Blinding.......................................................................................................33
6.5.
Packaging and Labeling..............................................................................33
6.6.
Preparation/Handling/Storage/Accountability..............................................33
6.7.
Compliance with Study Treatment Administration.......................................34
6.8.
Treatment of Study Treatment Overdose....................................................34
6.9.
Treatment after the End of the Study ..........................................................34
6.10.
Lifestyle and/or Dietary Restrictions............................................................34
6.10.1.
Meals and Dietary Restrictions ....................................................34
6.10.2.
Caffeine, Alcohol, and Tobacco ...................................................35
6.10.3.
Activity .........................................................................................35
6.11.
Concomitant Medications and Non-Drug Therapies....................................35
6.11.1.
Permitted Medications and Non-Drug Therapies..........................35
6.11.2.
Prohibited Medications and Non-Drug Therapies.........................35
 
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7.
STUDY ASSESSMENTS AND PROCEDURES .....................................................36
7.1.
Time and Events Table...............................................................................37
7.2.
Screening and Critical Baseline Assessments ............................................38
7.2.1.
Demographic/Medical History Assessments ................................38
7.3.
Pharmacokinetics .......................................................................................38
7.3.1.
Blood Sample Collection..............................................................38
7.3.2.
Sample Analysis ..........................................................................38
7.4.
Safety .........................................................................................................39
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)..........39
7.4.1.1.
Time period and Frequency for collecting AE 
and SAE information...................................................39
7.4.1.2.
Method of Detecting AEs and SAEs ...........................40
7.4.1.3.
Follow-up of AEs and SAEs........................................40
7.4.1.4.
Regulatory Reporting Requirements for SAEs............40
7.4.2.
Physical Exams ...........................................................................40
7.4.3.
Vital Signs....................................................................................41
7.4.4.
Electrocardiogram (ECG).............................................................41
7.4.5.
Clinical Safety Laboratory Assessments ......................................41
8.
DATA MANAGEMENT ...........................................................................................43
9.
STATISTICAL CONSIDERATIONS AND DATA ANALYSES .................................43
9.1.
Hypotheses.................................................................................................43
9.2.
Sample Size Considerations.......................................................................43
9.2.1.
Sample Size Assumptions ...........................................................43
9.2.2.
Sample Size Sensitivity................................................................44
9.2.3.
Sample Size Re-estimation or Adjustment...................................44
9.3.
Data Analysis Considerations .....................................................................44
9.3.1.
Analysis Populations....................................................................45
9.3.1.1.
Safety Population .......................................................45
9.3.1.2.
Pharmacokinetic Concentration Population ................45
9.3.1.3.
Pharmacokinetic Parameter Population......................45
9.3.2.
Interim Analysis ...........................................................................45
9.4.
Key Elements of Analysis Plan ...................................................................45
9.4.1.
Pharmacokinetic Analyses...........................................................45
9.4.2.
Safety Analyses...........................................................................46
10. STUDY GOVERNANCE CONSIDERATIONS........................................................46
10.1.
Posting of Information on Publicly Available Clinical Trial Registers............46
10.2.
Regulatory and Ethical Considerations, Including the Informed 
Consent Process ........................................................................................46
10.3.
Quality Control (Study Monitoring) ..............................................................47
10.4.
Quality Assurance.......................................................................................47
10.5.
Study and Site Closure ...............................................................................48
10.6.
Records Retention......................................................................................48
10.7.
Provision of Study Results to Investigators, Posting of Information 
on Publically Available Clinical Trials Registers and Publication .................49
11. REFERENCES.......................................................................................................50
12. APPENDICES........................................................................................................52
12.1.
Appendix 1: Abbreviations and Trademarks................................................52
 
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12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments ..............................................................................................53
12.3.
Appendix 3: Definition of and Procedures for Recording, Evaluating, 
Follow-Up and Reporting of Adverse Events...............................................55
12.3.1.
Definition of Adverse Events........................................................55
12.3.2.
Definition of Serious Adverse Events...........................................56
12.3.3.
Definition of Cardiovascular Events .............................................57
12.3.4.
Recording of AEs and SAEs ........................................................58
12.3.5.
Evaluating AEs and SAEs............................................................58
12.3.6.
Reporting of SAEs to GSK...........................................................60
12.4.
Appendix 4: High Fat Meal Content ............................................................61
12.5.
Appendix 5: Protocol Changes....................................................................62
 
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1.
PROTOCOL SYNOPSIS FOR STUDY 201964
Rationale
This study is designed to select one, or more fixed dose combinations (FDCs) of 
ambrisentan and tadalafil for further development and to provide pharmacokinetic data to 
enable a pivotal bioequivalence study (BE). Dependent on formulation work, the study 
will allow up to 8 new FDCs to be compared with the reference of ambrisentan and 
tadalafil monotherapies taken concurrently. The study would also allow for up to 2 of the 
new formulations, that may be taken in to a BE study, to be tested for any effect on 
pharmacokinetics of the FDC in both fed and fasted state.
Objective(s)/Endpoint(s)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following administration 
of candidate tablet formulations of the FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), and 
AUC(0-t) of ambrisentan and tadalafil in FDC 
and reference treatments
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
Overall Design
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects.
All subjects will attend the unit for Screening within 31 days of their first dose. Eligible 
subjects will be randomised to order of treatment in that study parts in which they are 
included and the order in which these treatments are administered will be in accordance 
with the randomisation schedule.
 
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There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose.  The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast.
Treatment Arms and Duration
The proposed treatment arms for each study part are described here; however the 
treatments in Parts 2 and 3 may be changed dependent on the utility and results from the 
previous part.
 
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Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil 
FDC5 (10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil 
FDC6 (10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil 
FDC7 (10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC4 (10mg/40mg)
ambrisentan and tadalafil 
FDC8 (10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less FDC treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. The minimum will be approximately 3 weeks and the maximum will be 
approximately 18 weeks. 
Type and Number of Subjects
A approximately of 20 healthy adult subjects will be randomized, to each study part, such 
that at least 16 evaluable subjects complete each part of the study.
Analysis
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and Part 2, (ii) estimate the bioavailability of the formulation(s) of 
the FDC formulations, taken in Part 2, if used for food effect and  Part 3, in the fed state 
relative to the fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
 
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2.
INTRODUCTION
Ambrisentan (E.U. trade name: Volibris), an orally active endothelin receptor antagonists 
(ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first approved on 
15 June 2007 in the US and on 21 April 2008 in the European Union (EU) and is 
currently approved in over 50 countries. In the EU, ambrisentan is indicated for treatment 
of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional Class 
(FC) II to III, including use in combination treatment (Volibris, EMA SmPC, 2015
Section 5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated 
with connective tissue disease
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (Adcirca, EMA, SmPC, 2015).
A recently completed study (Galiè, 2015a) has show that patients with PAH who started 
initial combination therapy with ambrisentan and tadalafil had a significantly lower risk 
of clinical-failure events compared to those that started with ambrisentan or tadalafil 
monotherapy. Ambrisentan has recently received EU approval (20 November 2015) for 
use in combination treatment with tadalafil (Volibris, EMA SmPC, 2015, Section 5.1). 
This pilot study will investigate the relative bioavailability of new fixed dose 
combinations (FDC) of ambrisentan and tadalafil, compared to the two monotherapies 
taken concurrently in healthy subjects
2.1.
Study Rationale
This study is designed to select one, or more, FDCs of ambrisentan and tadalafil for 
further development and to provide pharmacokinetic data to enable a pivotal 
bioequivalence study (BE). Dependent on formulation work, the study will allow up to 8 
new FDCs to be compared with the reference of ambrisentan and tadalafil monotherapies 
taken concurrently. The study would also allow for up to 2 of the new formulations, that 
may be taken in to a BE study, to be tested for any effect on pharmacokinetics of the 
FDC in both fed and fasted state.
The formulation(s) to be taken forward into the BE study will be primarily chosen on the 
test/reference ratio for both AUC and Cmax for both components ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, between subject variability, 
tablet size, cost, ease of manufacture and stability would be considered. 
2.2.
Brief Background
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
 
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biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galiè, 2015b) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris, EMA SmPC, 2015 Section 5.1). Efficacy has been 
shown in idiopathic PAH (IPAH) and in PAH associated with connective tissue 
disease.(Volibris, EMA SmPC,2015). Ambrisentan is an oral, once daily, propanoic acid-
based, ETA-selective Endothelin receptor antagonist (ERA) which targets the 
phospholipase-C-dependent endothelin pathway and which is known to play an essential 
role in mammalian cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (Adcirca, EMA, SmPC, 2015). 
Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) inhibitor which targets 
the NO pathway. Through inhibition of PDE-5, tadalafil increases cytoplasmic cGMP 
concentrations in the smooth muscle cells and enhances NO-mediated vasodilatation of 
the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112, 2008) in 26 healthy subjects was performed to detect 
any significant PK interactions between tadalafil and ambrisentan when co-administered 
(Spence, 2009). From this study, it was concluded that there is no clinically significant 
PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when combined. 
Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
administered. . There were no SAE’s in the study. Three subjects withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one subject because of myalgia (severe), 
muscle fatigue and dizziness on tadalafil alone and one because of headache (severe) on 
the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild and is a 
 
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listed event for ambrisentan. There were a total of 7 subjects with mild tachycardia. There 
were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. There were 3 events on 
the first day of tadalafil 40 mg given 3 days after ambrisentan 10 mg. There was one 
event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of ambrisentan 10 mg. 
Taken together these data suggest that in healthy subjects a mild tachycardia may result 
from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, EMA SmPC, 
2015, Adcirca, EMA, SmPC, 2015).
The AMBITION clinical study (Galiè, 2015a and AMB112565 CSR 13Nov2014), which 
evaluated the time to first clinical failure event, a composite endpoint, shows a robust 
clinical benefit (50% hazard reduction) for PAH patients initiated on a combination of 
ambrisentan and tadalafil when compared to PAH patients initiated on either medication 
as monotherapy.  The safety profile of the combination arm was consistent with the 
known safety data of the individual study drugs and no safety signals specific to 
combination treatment were identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galiè, 
2015b) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
and shipment requirements. This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 
CHMP/EWP/240/95) (2008).
 
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3.
OBJECTIVE(S) AND ENDPOINT(S)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following 
administration of candidate tablet formulations of 
the FDC (ambrisentan 10 mg + tadalafil 40 mg) 
relative to reference monotherapies tested 
(ambrisentan 10 mg & tadalafil 40 mg) taken 
concurrently in healthy human subjects under 
fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), 
and AUC(0-t) of ambrisentan and tadalafil in 
FDC and reference treatments
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2
of ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of 
adverse events
4.
STUDY DESIGN
4.1.
Overall Design
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects. See Figure 1 for study schematic.
 
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Figure 1
Study Schematic
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose. 
The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
 
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monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast.
Subjects who had taken part in the study part preceding Part 3, and which provided the 
successful formulations for a BE study could also be invited to take part in this study 
part. However, a subject’s inclusion in more than one study part would be dependent on 
the subject not exceeding the maximum blood draw volume (500ml) for the study.
4.2.
Treatment and Duration
The treatments for each study part of the study are listed in Table 1 All treatments are 
single dose. Subjects will be randomised to order of treatments in the parts of the study 
they are included in.
The study has 3 parts and ongoing analyse of pharmacokinetic data will be used to enable 
the formulations produced and tested in subsequent parts.
 
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Part 1
Part 1 of the study will be utilised to look at up to 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2. 
If no successful formulations are identified from these criteria then the pharmacokinetic 
data would be used to enable further work to produce new FDC formulations for Part 2
Following completion of Part 1 there will be a pause prior to Part 2, so that up to a further 
4 FDC formulations could be produced and data included and approved in any required 
update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for up to 4 FDC formulations. These could be 
either successful formulation identified in Part 1 reformulated with the final API and/or 
new formulations using the final API. Pharmacokinetic data from Part 2 of the study will 
be analysed following completion of each treatment arm by subjects. This ongoing 
review of the pharmacokinetic data will enable rapid ongoing assessment of each FDC 
formulation and will be used to define any successful FDC formulation to be taken into 
Part 3 of this study and a pivotal BE study. If only two formulations are evaluated in Part 
2 then the food effect may be added to Part 2 and Part 3 will not be required. Success will 
be defined with the same criteria as those in Part 1.
Part 3
Part 3 of the study will provide data for up to 2 successful formulations to be progressed 
to a pivotal BE study, in both the fed and fasted state.
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil 
FDC5 (10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil 
FDC6 (10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil 
FDC7 (10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC4 (10mg/40mg)
ambrisentan and tadalafil 
FDC8 (10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. Subjects may be included in Part 1, Part 2 or Part 3 of the study. However, if Part 2 
of the study identifies any successful formulations to take into Part 3 then subjects from 
Part 2 may be invited to join Part 3.
The minimum and maximum duration for a subject on this study is described in Table 2. 
This table assumes all 5 dose sessions are used for Part 1 and Part 2 and either 2 or 4 dose 
sessions are used in Part 3. However, this is an approximation as fewer sessions may be 
required in Part 2 and/or the washout between sessions of 7 days in each part may be 
longer, due to logistical reasons.
 
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Table 2
Study Duration
Study Options
Screen
Study 
Part 
1or 2
Pause
Study 
part 3
Follow-
Up 
Visit
Total 
Duration 
on Study
(1-31 
days)
(31 
days)
(7-28 
days)
(10-24 
days)
(7-14 
days)
Subjects 
included 
in only 
Part 1 
or 2
Min
1
31
7
39
Max
31
31
14
76
Subjects 
included 
in Part 
2&3
Min
1
31
7
10
7
56
Max
31
31
28
24
14
128
Subjects 
included 
in only 
Part 3
Min
1
10
7
18
Max
31
24
14
69
A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
4.3.
Type and Number of Subjects
A approximately of 20 healthy adult subjects will be randomized, to each study part, such 
that at least 16 evaluable subjects complete each part of the study.
The study is not powered. However, the number of subjects will be sufficient for the 
objectives of this study and this is detailed further in Section 9.2 Sample Size 
Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
 
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4.4.
Design Justification
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations to take into a pivotal BE study and to also provide pharmacokinetic data to 
enable the pivotal BE study to be powered sufficiently. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, or is there any need to blind study treatment. The Inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study is in three parts to enable PK data from Part 1 to enable formulation 
development for Part 2. Part 3 providing an opportunity to assess food effect in any FDC 
formulations suitable for development and inclusion in the pivotal bioequivalence study. 
The interdependency of the 3 study parts is described in detail in Section 4.2. 
4.5.
Dose Justification
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of, 40mg for tadalafil and 10mg for ambrisentan are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to 
be tolerated in healthy subjects.
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimum washout of 7 days, 
between doses, is also sufficient for clearance of previous dose.
4.6.
Benefit: Risk Assessment
There is no direct benefit for healthy subjects participating in this study.  The risks to the 
healthy subjects based upon previous experience indicate no expectation of SAE and mild 
to moderate AE.  The benefit/risk remains satisfactory for this healthy subject study.
Summaries of findings from clinical studies conducted with both the Investigational 
Product taken as monotherapies and in combination can be found in: 

Ambrisentan IB (Volibris IB, 2015)

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC, 2015) 

Adcirca (Tadalafil) EMA SmPC (Adcirca, EMA SmPC, 2015)
 
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As described in Section 2.2, mild tachycardia was the most common (7/26, 27%) adverse 
event seen in healthy subjects following combination dosing with 10mg ambrisentan and 
40mg tadalafil (Spence, 2009). There were three subjects (3/26, 11.5%) withdrawn due to 
AEs (e.g., anaemia [mild]; myalgia [severe], muscle fatigue, and dizziness; and headache 
[severe]).    No SAEs or additional risks were observed with combination use in healthy 
subjects.
The following section outlines the risk assessment and mitigation strategy for this 
protocol:  
4.6.1.
Risk Assessment
Risks of clinical significance, identified in subjects with PAH and the mitigation strategy 
identified for this study in healthy subjects are captured in Table 3. 
 
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Table 3
Risks of Clinical Significance and Mitigation Strategy
Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
Investigational Product: GSK1325760 (ambrisentan-Volibris)

Teratogenicity

Anaemia (decreased 
haemoglobin, decreased 
haematocrit)

Hypersensitivity reactions 
(e.g. angioedema, rash, 
pruritus)

Headache (including sinus 
headache, migraine)

Dizziness

Cardiac failure

Palpitation

Hypotension

Flushing

Epistaxis

Dyspnoea

Upper respiratory  
congestion, sinusitis, 
nasopharyngitis, rhinitis

Abdominal pain

Constipation

Nausea, vomiting, 
diarrhoea

Hepatic transaminases 
increased

Peripheral oedema, fluid 
retention

Chest pain/discomfort

Asthenia and fatigue
Ambrisentan IB (Volibris IB, 
2015)
Volibris (Ambrisentan) EMA 
SmPC (Volibris, EMA SmPC, 
2015): 
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision. 

Exclusion of women of 
childbearing potential 
(teratogenicity)

Laboratory assessments 
per protocol

Physical assessment per 
protocol

Routine vital signs per 
protocol

Subjects remain in the 
clinical unit, under medical 
supervision, for all doses 
and until completion of 
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GF196960 (tadalafil - Adcirca)

Hypersensitivity reactions

Headache

Syncope,

Migraine

Blurred vision

Palpitations

Nasopharyngitis (including 
nasal congestion, sinus 
congestion and rhinitis)

Epistaxis
Adcirca (Tadalafil) EMA SmPC 
(Adcirca, EMA SmPC, 2015))
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision.

Routine vital signs per 
protocol

Physical assessment per 
protocol

Subjects remain in the 
clinical unit for all doses, 
under medical supervision 
and until completion of 
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy

Nausea,

Dyspepsia (including 
abdominal pain/discomfort)

Vomiting,

Gastroesophageal reflux

Rash

Myalgia,

Back pain

Pain in extremity (including 
limb discomfort)

Increased uterine bleeding

Facial oedema,

Chest pain
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GSK3380154 (ambrisentan-tadalafil-FDC)
The risks for the combination 
are the same as the 2 
monotherapies; no extra risks 
have been identified for the 
combination.
As for GSK1325760 and 
GF196960 above
4.6.2.
Benefit Assessment
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation for a 
fixed dose combination of ambrisentan plus tadalafil to use as first line therapy in PAH 
patients.
4.6.3.
Overall Benefit: Risk Conclusion
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 3. So, the 
risk: benefit is appropriate for this study and to enable development of this investigational 
product. The overall benefit: risk therefore remains positive.
5.
SELECTION OF STUDY POPULATION AND 
WITHDRAWAL CRITERIA
Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in:

Volibris (Ambrisentan) EMA SmPC (Volibris, EMA SmPC, 2015)

Adcirca (Tadalafil) EMA SmPC (Adcirca, EMA SmPC, 2015)
 
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Deviations from inclusion and exclusion criteria are not allowed because they can 
potentially jeopardize the scientific integrity of the study, regulatory acceptability or 
subject safety. Therefore, adherence to the criteria as specified in the protocol is essential.
5.1.
Inclusion Criteria
A subject will be eligible for inclusion in this study only if all of the following criteria 
apply:
AGE
1. Between 18 and 60 years of age inclusive, at the time of signing the informed 
consent.
TYPE OF SUBJECT AND DIAGNOSIS INCLUDING DISEASE SEVERITY
2. Healthy as determined by a responsible and experienced physician, based on a 
medical evaluation including medical history, physical examination, laboratory tests, 
vital signs and cardiac monitoring (ECG and 24 hour Holter).  A subject with a 
clinical abnormality or laboratory parameter(s) which is/are not specifically listed in 
the inclusion or exclusion criteria, outside the reference range for the population 
being studied may be included only if the Investigator, in consultation with the GSK 
Medical Monitor if required, judges and documents that the finding is unlikely to 
introduce additional risk factors and will not interfere with the study procedures. 
WEIGHT
3. Body weight ≥ 50 kg (110 lbs) for men and ≥ 45 kg (99lbs) for women and body 
mass index (BMI) within the range 18 – 30 kg/m2 (inclusive)
SEX
4. Male or Female
Females must be the following:
Non-reproductive potential defined as:

Pre-menopausal females with one of the following:

Documented tubal ligation

Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion 

Hysterectomy

Documented Bilateral Oophorectomy

Documented Postmenopausal defined as 12 months of spontaneous amenorrhea
 
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INFORMED CONSENT
5. Capable of giving signed informed consent as described in Section 10 which includes 
compliance with the requirements and restrictions listed in the consent form and in 
this protocol.
5.2.
Exclusion Criteria
A subject will not be eligible for inclusion in this study if any of the following criteria 
apply:
CONCURRENT CONDITIONS/MEDICAL HISTORY (INCLUDES LIVER 
FUNCTION AND QTC INTERVAL)
1.
A blood pressure <100/55 mm Hg.
2.
Haemoglobin below normal range:

Hb < 133 g/L for males 

Hb< 114 g/L for females
3.
ALT and bilirubin >1.5xULN (isolated bilirubin >1.5xULN is acceptable if bilirubin 
is fractionated and direct bilirubin <35%). 
4.
Current or chronic history of liver disease, or known hepatic or biliary abnormalities 
(with the exception of Gilbert's syndrome or asymptomatic gallstones)
5.
QTc> 450 msec
NOTES:

The QTc is the QT interval corrected for heart rate according to Bazett’s 
formula (QTcB), Fridericia’s formula (QTcF), and/or another method, machine-
read or manually over-read.

The specific formula that will be used to determine eligibility and 
discontinuation for an individual subject should be determined prior to initiation 
of the study. In other words, several different formulae cannot be used to 
calculate the QTc for an individual subject and then the lowest QTc value used 
to include or discontinue the subject from the trial.

For purposes of data analysis, QTcB, QTcF, another QT correction formula, or a 
composite of available values of QTc will be used as specified in the Reporting 
and Analysis Plan (RAP).
CONCOMITANT MEDICATIONS
6.
Use of prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements (including St John’s Wort) within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose 
 
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of study medication, unless in the opinion of the Investigator and GSK Medical 
Monitor the medication will not interfere with the study procedures or compromise 
subject safety. 
RELEVANT HABITS
7.
History of regular alcohol consumption within 6 months of the study defined as: 

An average weekly intake of >21 units for males or >14 units for females.  One 
unit is equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 
ml) of wine or 1 (25 ml) measure of spirits.
8.
Smoking more than 5 cigarettes per week and subjects must be able to abstain from 
smoking for a 24 hour period prior to dose and any time whilst in the clinical unit.
CONTRAINDICATIONS
9.
History of sensitivity to any of the study medications, or components thereof or a 
history of drug or other allergy that, in the opinion of the Investigator or Medical 
Monitor, contraindicates their participation.
DIAGNOSTIC ASSESSMENTS AND OTHER CRITERIA
10. Presence of hepatitis B surface antigen (HBsAg), positive hepatitis C antibody test 
result at Screening or within 3 months prior to first dose of study treatment. . 
11. A positive test for HIV antibody.
12. A positive pre-study drug/alcohol screen.  
13. Where participation in the study would result in donation of blood or blood products 
in excess of 500 mL within previous 3 months
14. The subject has participated in a clinical trial and has received an investigational 
product within the following time period prior to the first dosing day in the current 
study: 3 months, 5 half-lives or twice the duration of the biological effect of the 
investigational product (whichever is longer).
15. Exposure to more than four new chemical entities within 12 months prior to the first 
dosing day.
5.3.
Screening/Baseline/Run-in Failures
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT) 
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
 
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set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.4).
5.4.
Withdrawal/Stopping Criteria
The following actions must be taken in relation to a subject who fails to attend the clinic 
for a required study visit:

The site must attempt to contact the subject and re-schedule the missed visit as 
soon as possible. 

The site must counsel the subject on the importance of maintaining the assigned 
visit schedule and ascertain whether or not the subject wishes to and/or should 
continue in the study. 

In cases where the subject is deemed ‘lost to follow up’, the investigator or 
designee must make every effort to regain contact with the subject (where 
possible, 3 telephone calls and if necessary a certified letter to the subject’s last 
known mailing address or local equivalent methods). These contact attempts 
should be documented in the subject’s medical record. 

Should the subject continue to be unreachable, only then will he/she be considered 
to have withdrawn from the study with a primary reason of “Lost to Follow-Up”. 
A subject may withdraw from study treatment at any time at his/her own request, or may 
be withdrawn at any time at the discretion of the investigator for safety, behavioural or 
administrative reasons. If a subject withdraws from the study, he/she may request 
destruction of any samples taken, and the investigator must document this in the site 
study records. 
5.4.1.
Liver Chemistry Stopping Criteria
Liver chemistry stopping and increased monitoring criteria have been designed to 
assure subject safety and evaluate liver event etiology (in alignment with the FDA 
premarketing clinical liver safety guidance). 
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf
 
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Study treatment will be discontinued for a subject if liver chemistry stopping criteria are 
met:
Phase I Liver Chemistry Stopping Criteria – Liver Stopping Event Algorithm
Liver Safety Required Actions and Follow-Up Assessments Section can be found in 
Appendix 2: Liver Safety Required Actions and Follow-Up Assessments
5.4.1.1.
Study Treatment Restart or Rechallenge
Study treatment restart or re-challenge after liver chemistry stopping criteria are met by 
any subject participating in this study is not allowed.
5.4.2.
QTc Stopping Criteria

The same QT correction formula must be used for each individual subject to 
determine eligibility for and discontinuation from the study.  This formula may 
not be changed or substituted once the subject has been enrolled.

For example, if a subject is eligible for the protocol based on QTcF, then QTcF 
must be used for discontinuation of this individual subject as well.

Once the QT correction formula has been chosen for a subject’s eligibility, the 
same formula must continue to be used for that subject for all QTc data being 
collected for data analysis.  Safety ECGs and other non-protocol specified 
ECGs are an exception.  

The QTc should be based on averaged QTc values of triplicate electrocardiograms 
obtained over a brief (e.g., 5-10 minute) recording period. 
A subject that meets either bulleted criterion below will be withdrawn from the study.  

QTc> 500 msec,
Continue Study Treatment 
Discontinue Study Treatment 
*INR value not applicable to subjects on anticoagulants 
ALT ≥3xULN 
No
Yes
Must refer to Liver Safety Required Actions and Follow up Assessments  section in the Appendix
Report as an SAE if possible Hy’s Law case: ALT≥3xULN and  Bilirubin≥2xULN (>35% direct)  or 
INR>1.5, if measured*
 
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
Change from baseline: QTc >60 msec 
5.4.3.
Hypotension
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 mmHG and diastolic <50 mm HG confirmed by 
triplicate reading taken up to 5 minutes apart and is judged clinically significant 
and symptomatic by the investigator.
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria
For an individual study participant, stopping criteria include, but are not limited to:
Adverse events, or significant changes in any of the safety assessments, that put the 
safety of the individual at risk (e.g., ECG, vital signs, laboratory tests, etc), as judged by 
the Principal Investigator in consultation with the Medical Monitor if necessary.
5.5.
Subject and Study Completion
A completed subject is one who has completed all phases of the study including the 
Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
6.
STUDY TREATMENT
6.1.
Investigational Product and Other Study Treatment
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Part 1 of the study are shown in Table 4, this will be 
amended once treatments for Part 2 and Part 3 are confirmed.
 
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Table 4
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS*
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS*
*
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 95 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 233 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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6.2.
Medical Devices
No GSK manufactured devices (or devices manufactured for GSK by a third party) are 
provided for use in this study.
6.3.
Treatment Assignment
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software.
The treatments are denoted as F1 to F4 for Part 1, F5 to F8 for Part 2 and R for reference 
treatment for both parts; the selected formulations for Part 3 are denoted as X and Y.  The 
treatment key for Parts 1, 2 and 3 are described in Table 5
Table 5
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 2
F5
ambrisentan and tadalafil FDC5 (10mg/40mg)
F6
ambrisentan and tadalafil FDC6 (10mg/40mg)
F7
ambrisentan and tadalafil FDC7 (10mg/40mg)
F8
ambrisentan and tadalafil FDC8 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3
X1
ambrisentan and tadalafil FDCX1 (10mg/40mg), fed
X2
ambrisentan and tadalafil FDCX1 (10mg/40mg) fasted
Y1
ambrisentan and tadalafil FDCX2 (10mg/40mg), fed
Y2
ambrisentan and tadalafil FDCX2 (10mg/40mg), fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 6. Not all possible sequences are 
included here. Such as, if only one formulation taken into Part 2 or 3, or Part 2 is used for 
 
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only one or two FDC formulations, fed and fasted. Additional treatment sequences will 
be created based on the Latin Squares for Williams Designs, as required.
Table 6
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1 F2 R F3 F4
F2 F3 F1 F4 R
F3 F4 F2 R F1
F4 R F3 F1 F2
R F1 F4 F2 F3
F4 F3 R F2 F1
R F4 F1 F3 F2
F1 R F2 F4 F3
F2 F1 F3 R F4
F3 F2 F4 F1 R
1:1:1:1:1:1:1:1:1:1
Part 2
5
F5 F6 R F7 F8
F6 F7 F5 F8 R
F7 F8 F6 R F5
F8 R F7 F5 F6
R F5 F8 F6 F7
F8 F7 R F6 F5
R F8 F5 F7 F6
F5 R F6 F8 F7
F6 F5 F7 R F8
F7 F6 F8 F5 R
1:1:1:1:1:1:1:1:1:1
4
F5 F6 R F7
F6 F7 F5 R
F7 R F6 F5
R F5 F7 F6
1:1:1:1
3
F5 F6 R
R F6 F5
F6 R F5
F1 R F6
R F5 F6
F6 F5 R
3:3:3:3:3:3 + 2 random
2
F5 R 
R F5
1:1
Part 3
4
X1 X2 Y2 Y1
X2 Y1 X1 Y2
Y1 Y2  X2 X1
Y2  X1 Y1 X2 
1:1:1:1
Part 2 and 
Part 3 
merged 
together
5 (Two FDCs selected from 
Part 1)
X1 X2 R Y1 Y2
X2 Y1 X1 Y2 R
Y1 Y2 X2 R X1
Y2 R Y1 X1 X2
R X1 Y2 X2 Y1
Y2 Y1 R X2 X1
1:1:1:1:1:1:1:1:1:1
 
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Study Parts
Number of Treatments
Sequence assignments
Allocation ratio
R Y2 X1 Y1 X2
X1 R X2 Y2 Y1
X2 X1 Y1 R Y2
Y1 X2 Y2 X1 R
3 (One FDC selected from 
Part 1)
X1 X2 R
R X2 X1
X2 R X1
F1 R X2
R X1 X2
X2 X1 R
3:3:3:3:3:3 + 2 random
6.4.
Blinding
This will be an open-label study. 
6.5.
Packaging and Labeling
The contents of the label will be in accordance with all applicable regulatory 
requirements.
6.6.
Preparation/Handling/Storage/Accountability
This will be detailed in a Study Specific Technical Agreement/Memo (TTS) which will 
be accompanied by a Quality Agreement.

Only subjects enrolled in the study may receive study treatment and only 
authorized site staff may supply or administer study treatment.  All study 
treatments must be stored in a secure environmentally controlled and monitored 
(manual or automated) area in accordance with the labelled storage conditions 
with access limited to the investigator and authorized site staff. 

The investigator, institution, or the head of the medical institution (where 
applicable) is responsible for study treatment accountability, reconciliation, and 
record maintenance (i.e. receipt, reconciliation and final disposition records).  

Further guidance and information for final disposition of unused study treatment 
are provided in the SRM. 

Under normal conditions of handling and administration, study treatment is not
expected to pose significant safety risks to site staff.  

A Material Safety Data Sheet (MSDS)/equivalent document describing 
occupational hazards and recommended handling precautions either will be 
provided to the investigator, where this is required by local laws, or is available 
upon request from GSK.
 
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6.7.
Compliance with Study Treatment Administration
When subjects are dosed at the site, they will receive study treatment directly from the 
investigator or designee, under medical supervision.  The date and time of each dose 
administered in the clinic will be recorded in the source documents.  The dose of study 
treatment and study subject identification will be confirmed at the time of dosing by a 
member of the study site staff other than the person administering the study treatment.  
Study site personnel will examine each subject’s mouth to ensure that the study treatment 
was ingested.
6.8.
Treatment of Study Treatment Overdose
For this study, any dose of GSK3380154 (ambrisentan/tadalafil-FDC), GSK1325760 
(ambrisentan) and GF196960 (tadalafil) greater than the protocol defined dose and within 
a 24 hour time period will be considered an overdose.
GSK does not recommend specific treatment for an overdose.
Advice for the investigator is included in the product label for GSK1325760 
(ambrisentan) and GF196960 (tadalafil).
In the event of an overdose the investigator should:
1.
Contact the Medical Monitor immediately
2.
Closely monitor the subject for adverse events (AEs)/serious adverse events (SAEs) 
and laboratory abnormalities until compound number/name can no longer be 
detected systemically (at least 3 days for compound number/name)
3.
Obtain a plasma sample for pharmacokinetic (PK) analysis if requested by the 
Medical Monitor (determined on a case-by-case basis)
4.
Document the quantity of the excess dose as well as the duration of the overdosing in 
the CRF.
Decisions regarding dose interruptions or modifications will be made by the investigator 
in consultation with the Medical Monitor based on the clinical evaluation of the subject.
6.9.
Treatment after the End of the Study
Subjects will not receive any additional treatment from GSK after completion of the 
study because only healthy subjects are eligible for study participation.
6.10.
Lifestyle and/or Dietary Restrictions
6.10.1.
Meals and Dietary Restrictions

Refrain from consumption of red wine, Seville oranges, grapefruit or grapefruit 
juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or fruit juices from 
7 days prior to the first dose of study medication until after the final dose.
 
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
Dependent on the utility of the dose session subjects will be asked to fast or 
consume the FDA full fat breakfast prior to dosing:

Fasting subjects will be required to fast from midnight before each full in-house 
dosing day (i.e. Day 1) with the exception of water, which will be allowed freely 
except for 1 hour either side of dosing. Subjects will be required to fast up to 4h 
hour post dose on Day 1 of each dose

For treatments given under fed conditions, a standard high-fat breakfast will be 
provided before dosing. See Section 12.4 for details of this meal. This meal 
should be eaten in its entirety within 30 minutes.  The amount consumed will be 
recorded within the source documents and the CRF by the site staff.  Subjects 
will be dosed within 5 minutes of completing the breakfast.  

Subjects should take each dose of investigational product with 240 ml (8 fl oz) of 
water.
6.10.2.
Caffeine, Alcohol, and Tobacco

During each dosing session, subjects will abstain from ingesting caffeine- or 
xanthine-containing products (e.g., coffee, tea, cola drinks and chocolate) for 24 
hours prior to the start of dosing until collection of the final pharmacokinetic sample 
during each session. 

During each dosing session, subjects will abstain from alcohol for 24 hours prior to 
the start of dosing until collection of the final pharmacokinetic sample during each 
session. 

Smoking is not allowed for 24 hours prior to dosing and whilst subjects are in the 
clinic.
6.10.3.
Activity
Subjects will abstain from strenuous exercise from Screening until Follow-Up.  Subjects 
may participate in light recreational activities during studies (e.g., watch television, read).
6.11.
Concomitant Medications and Non-Drug Therapies
6.11.1.
Permitted Medications and Non-Drug Therapies
Paracetamol at doses of 2 grams/day is permitted for use any time during the study. 
Other concomitant medication may be considered on a case by case basis by the 
Investigator in consultation with the Medical Monitor if required.
6.11.2.
Prohibited Medications and Non-Drug Therapies
Subjects must abstain from taking prescription or non-prescription drugs (including 
vitamins and dietary or herbal supplements), within 7 days (or 14 days if the drug is a
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose of 
study medication until completion of the Follow-Up visit, unless in the opinion of the 
Investigator and sponsor the medication will not interfere with the study.
 
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7.
STUDY ASSESSMENTS AND PROCEDURES
Protocol waivers or exemptions are not allowed with the exception of immediate safety 
concerns. Therefore, adherence to the study design requirements, including those 
specified in the Time and Events Table, are essential and required for study conduct.
This section lists the procedures and parameters of each planned study assessment.  The 
exact timing of each assessment is listed in the Time and Events Table Section 7.1
The following points must be noted: 

If assessments are scheduled for the same nominal time, THEN the assessments 
should occur in the following order:
1. 12-lead ECG
2. Vital signs
3. Blood draws.  
Note: The timing of the assessments should allow the blood draw to occur at the 
exact nominal time.

The timing and number of planned study assessments, safety and 
pharmacokinetic assessments may be altered during the course of the study 
based on newly available data (e.g., to obtain data closer to the time of peak 
plasma concentrations) to ensure appropriate monitoring. 

The change in timing or addition of time points for any planned study 
assessments must be documented in a Note to File which is approved by the 
relevant GSK study team member and then archived in the study sponsor and 
site study files, but this will not constitute a protocol amendment. 

The IRB/IEC will be informed of any safety issues that require alteration of the 
safety monitoring scheme or amendment of the Informed Consent Form. 
No more than 500 mL of blood will be collected over the duration of the study, including 
any extra assessments that may be required.
 
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7.1.
Time and Events Table
Procedure
Screen
Part 1, 2 and 3. Each dose in each Part repeats this schedule.
FU
Notes
Day
≤-31
-1
1
2
3
4
≥7-14
Time (hrs)
Pre-
dose
0
0.5
1
1.5
2
2.5
4
8
12
18
24
36
48
72
Outpatient visit
x
x
x
Admission to unit
x
Informed consent
x
Inclusion and exclusion criteria
x
x
Demography
x
Full physical exam including 
height and weight
x
Brief Physical
x
Medical history (includes 
substance usage)
x
HIV, Hep B and Hep C screen] 
x
Laboratory assessments (include 
liver chemistries) 
x
x
x
x
Only Screening labs need to be taken in 
fasted state.
Serum  hCG Pregnancy test
x
x
Female subjects only
Urine  hCG Pregnancy test
x
Female subjects only
Breathalyser and Smokerlyzer
x
x
DOA testing
x
x
12-lead ECG
x
x
x
x
x
x
x
x
x
Triplicate at screen and baseline, single 
measure at other times, unless out of 
range then triplicates should be 
performed
Vital signs
x
x
x
x
x
x
x
x
x
x
x
x
24hr Holter
x
Randomisation
x
Randomised prior to first dose only
Study Treatment
x
AE/SAE review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
SAEs from Screen. AEs from first dose
Concomitant medication review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
PK Sample
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Discharge from Unit
x
For logistical reasons subjects may 
remain in-unit for the 72 hr assessments 
if they prefer.
 
 
 
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7.2.
Screening and Critical Baseline Assessments
All subjects must give written consent to participate in this trial.  Consent for screening 
evaluations may be obtained using the ICF for the Hammersmith Medicines Research 
(HMR) healthy subject’s panel, which has been approved by the HRA’s Phase 1 Advert 
Review group.  The study-specific information and consent form will be signed by the 
subject either before any screening evaluation or after the investigator confirms the 
eligibility of the subject for the study and before the subject is randomised to receive the 
first administration of IMP.  Before giving consent, subjects must read the information 
sheet about the study.  They must also read the consent form.  They will then discuss the 
study with the investigator or his deputy and be given the opportunity to ask questions.  
The study-specific information sheet and the consent form must be approved by the REC.
7.2.1.
Demographic/Medical History Assessments
Prior to enrolling in the study and having any study procedures completed, subjects must 
sign the informed consent.
During the Screening visit, each subject will undergo the assessments to determine 
eligibility for enrolment as detailed in Section 5. 
The following demographic parameters will be captured: date of birth, gender, race and 
ethnicity.
Medical/medication/alcohol history will be assessed as related to the eligibility criteria 
listed in Section 5.
7.3.
Pharmacokinetics
7.3.1.
Blood Sample Collection
Blood samples for pharmacokinetic (PK) analysis of ambrisentan and tadalafil will be 
collected at the time points indicated in Section 7.1, Time and Events Table.  The actual 
date and time of each blood sample collection will be recorded.  The timing of PK 
samples may be altered and/or PK samples may be obtained at additional time points to 
ensure thorough PK monitoring.
For analysis of ambrisentan 2.7 mL of blood will be collected into sodium citrate tubes 
and for analysis of tadalafil 2.0 mL of blood will be collected into K2-EDTA tubes. 
Processing, storage and shipping procedures are provided in the Study Reference Manual 
(SRM).
7.3.2.
Sample Analysis
Plasma analysis will be performed under the control of Platform Technology and Science 
In vitro/In vivo Translation (PTS IVIVT) and Third Party Resource, GlaxoSmithKline, 
the details of which will be included in the Study Reference Manual (SRM).
Concentrations of ambrisentan and tadalafil will be determined in plasma samples using 
 
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the currently approved bioanalytical methodology. Raw data will be archived at the 
bioanalytical site (detailed in the SRM).
Once the plasma has been analyzed for ambrisentan and tadalafil any remaining 
plasma may be analyzed for other compound-related metabolites and the results reported 
under a separate PTS-IVIVT, GlaxoSmithKline protocol.
7.4.
Safety
Planned time points for all safety assessments are listed in the Time and Events Table 
(Section 7.1).  Additional time points for safety tests such as vital signs, physical 
examinations and laboratory safety assessments may be added during the course of the 
study based on newly available data to ensure appropriate safety monitoring.
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)
The definitions of an AE or SAE can be found in Appendix 3: Definition of and 
Procedures for Recording, Evaluating, Follow-Up and Reporting of Adverse Events.
The investigator and their designees are responsible for detecting, documenting and 
reporting events that meet the definition of an AE or SAE. 
7.4.1.1.
Time period and Frequency for collecting AE and SAE information

Any SAEs assessed as related to study participation (e.g., protocol-mandated 
procedures, invasive tests, or change in existing therapy) or related to a GSK 
product will be recorded from the time a subject consents to participate in the 
study up to and including any follow up contact. 

AEs will be collected from the start of Study Treatment until the follow up contact 
(see Section 7.4.1.3), at the timepoints specified in the Time and Events Table 
(Section 7.1).

Medical occurrences that begin prior to the start of study treatment but after 
obtaining informed consent may be recorded on the Medical History/Current 
Medical Conditions section of the CRF.

All SAEs will be recorded and reported to GSK within 24 hours, as indicated in 
Section 12.3.

Investigators are not obligated to actively seek AEs or SAEs in former study 
subjects. However, if the investigator learns of any SAE, including a death, at any 
time after a subject has been discharged from the study, and he/she considers the 
event reasonably related to the study treatment or study participation, the 
investigator must promptly notify GSK.
NOTE: The method of recording, evaluating and assessing causality of AEs and SAEs 
plus procedures for completing and transmitting SAE reports to GSK are provided in 
Section 12.3.
 
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7.4.1.2.
Method of Detecting AEs and SAEs
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended 
and non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrence.  Appropriate questions include:

“How are you feeling?” 

“Have you had any (other) medical problems since your last visit/contact?”

“Have you taken any new medicines, other than those provided in this study, 
since your last visit/contact?
7.4.1.3.
Follow-up of AEs and SAEs
After the initial AE/SAE report, the investigator is required to proactively follow each 
subject at subsequent visits/contacts.  All SAEs, and non-serious AEs of special interest 
(as defined in Section 4.6.1) will be followed until resolution, until the condition 
stabilizes, until the event is otherwise explained, or until the subject is lost to follow up 
(as defined in Section 5.4). Further information on follow up procedures is given in Study 
Assessments and Procedures.
7.4.1.4.
Regulatory Reporting Requirements for SAEs
Prompt notification by the investigator to GSK of SAEs related to study treatment (even 
for non- interventional post-marketing studies) is essential so that legal obligations and 
ethical responsibilities towards the safety of subjects and the safety of a product under 
clinical investigation are met. 
GSK has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of a product under clinical investigation.  GSK will 
comply with country specific regulatory requirements relating to safety reporting to the 
regulatory authority, Institutional Review Board (IRB)/Independent Ethics Committee 
(IEC) and investigators.
Investigator safety reports are prepared for suspected unexpected serious adverse 
reactions according to local regulatory requirements and GSK policy and are forwarded 
to investigators as necessary.
An investigator who receives an investigator safety report describing a SAE(s) or other 
specific safety information (e.g., summary or listing of SAEs) from GSK will file it with 
the IB and will notify the IRB/IEC, if appropriate according to local requirements.
7.4.2.
Physical Exams

A complete physical examination will include, at a minimum, assessment of the 
Cardiovascular, Respiratory, Gastrointestinal and Neurological systems. Height 
and weight will also be measured and recorded. 

A brief physical examination will include, at a minimum assessments of the skin, 
lungs, cardiovascular system, and abdomen (liver and spleen).
 
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
Investigators should pay special attention to clinical signs related to previous 
serious illnesses
7.4.3.
Vital Signs

Vital signs will be measured in semi-supine position after 5 minutes rest and will 
include temperature, systolic and diastolic blood pressure and pulse rate and 
respiratory rate. 

Three readings of blood pressure and pulse rate will be taken at Screening and 
baseline (pre-dose). All subsequent assessments will be single measures, unless 
the subjects blood pressure or pulse rate has changed from baseline by >15% and 
then 2 further readings should be taken and recorded:

For triplicate readings:

All 3 readings will be recorded in the CRF

First reading should be rejected

Second and third readings should be averaged to give the measurement to 
be used and this will also be recorded in the CRF
7.4.4.
Electrocardiogram (ECG)

ECGs will be measured in semi-supine position after 5 minutes rest

Triplicate 12-lead ECGs will be obtained at Screening and baseline (predose). All 
subsequent assessments will be a single measures, unless withdrawal criteria are 
meet and in which case 2 further readings should be taken to confirm if 
withdrawal is required . An ECG machine that automatically calculates the heart 
rate and measures PR, QRS, QT, and QTc.  Refer to Section 5.4.2 for QTc 
withdrawal criteria and additional QTc readings that may be necessary.

24 hour continuous cardiac telemetry (Holter) will be performed at Screening.  
Full disclosures will be reviewed in detail and the review maintained as part of the 
subject’s source documents
7.4.5.
Clinical Safety Laboratory Assessments
All protocol required laboratory assessments, as defined in Table 7, must be conducted in 
accordance with the Laboratory Manual, and Protocol Time and Events Schedule.
Laboratory requisition forms must be completed and samples must be clearly labelled 
with the subject number, protocol number, site/centre number, and visit dat.  Reference 
ranges for all safety parameters will be provided to the site by the laboratory responsible 
for the assessments.
If additional non-protocol specified laboratory assessments are performed at the 
institution’s local laboratory and result in a change in subject management or are 
considered clinically significant by the investigator (e.g., SAE or AE or dose 
modification) the results must be recorded in the CRF.
 
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Refer to the SRM for appropriate processing and handling of samples to avoid duplicate 
and/or additional blood draws.
All study-required laboratory assessments will be performed by a local laboratory.
Haematology, clinical chemistry, urinalysis and additional parameters to be tested are 
listed in Table 7. 
Table 7
Protocol Required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Haematology
Platelet Count
RBC Indices:
WBC count with Differential:
RBC Count
MCV
Neutrophils
Hemoglobin
MCH
Lymphocytes
Hematocrit
MCHC
Monocytes
Eosinophils
Basophils
Clinical 
Chemistry 1
BUN
Potassium
AST (SGOT)
Total and direct bilirubin
Creatinine
Sodium
ALT (SGPT)
Total Protein
Glucose 
Calcium
Alkaline phosphatise
Albumin
Routine 
Urinalysis3
As a minimum, but not limited to the following tests and dependent on 
standard urinalysis dipstick used:

Specific gravity

pH, glucose, protein, blood and ketones by dipstick

Microscopic examination (if blood or protein is abnormal)
Other 
Screening 
Tests

HIV

Hepatitis B (HBsAg)

Hepatitis C (Hep C antibody)

Alcohol and drug screen (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines)

Serum  or urine hCG Pregnancy test2
NOTES :
1.
Details of Liver Chemistry Stopping Criteria and Required Actions and Follow-Up 
Assessments after liver stopping or monitoring event are given in Section 5.4.1 and 
Appendix 2
2.
Local urine testing will be standard for the protocol unless serum testing is required 
by local regulation or ethics committee.
3.
Routine Urinalysis results will not be databased, unless a result is out of range and 
clinically significant then it would be captured as an AE  
 
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All laboratory tests with values that are considered clinically significantly abnormal 
during participation in the study or within7 days after the last dose of study treatment 
should be repeated until the values return to normal or baseline.  If such values do not 
return to normal within a period judged reasonable by the investigator, the etiology
should be identified and the sponsor notified.
8.
DATA MANAGEMENT

Data will be double-entered into a clinical database management system 
(ClinPlus Version 3.3).

Management of clinical data will be performed in accordance with applicable 
HMR standards and data cleaning procedures to ensure the integrity of the data, 
e.g., removing errors and inconsistencies in the data.

Adverse events and concomitant medications terms will be coded using 
MedDRA (Medical Dictionary for Regulatory Activities) and an internal 
validated medication dictionary, GSKDrug.  

Original CRFs will be retained by GSK, while HMR will retain a copy. Subject 
initials will not be collected or transmitted to GSK according to GSK policy 
9.
STATISTICAL CONSIDERATIONS AND DATA 
ANALYSES
9.1.
Hypotheses
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and if used, Study part 2, (ii) estimate the bioavailability of the 
formulation(s) of the FDC formulations, taken in to Part 3, in the fed state relative to the 
fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
9.2.
Sample Size Considerations
No formal sample size calculation has been performed. Sample sizes chosen for each 
study part in this study is considered sufficient for exploratory analysis based on PK 
variability data from previous studies
9.2.1.
Sample Size Assumptions
The sample size assumptions are based on previously reported estimates of within subject 
CV for AUC(0-∞) and Cmax for ambrisentan (GS-US-300-0112, 2008) and tadalafil 
(Forgue, 2005). Table 8 summarizes the estimates of within subject CV for the primary 
endpoints AUC (0-∞) and Cmax.
 
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Table 8
Estimates of within subject CV for the primary end points AUC (0-∞) 
and Cmax
CVw: within subject CV
ambrisentan tadalafil 
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
9.2.2.
Sample Size Sensitivity
Assuming the within subject CV differs from 22% and the number of statistically 
evaluable subjects is lower than 20, then Table 9 shows the precision (the half width of 
the 90% confidence interval) for a range of evaluable subjects.
Table 9
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
9.2.3.
Sample Size Re-estimation or Adjustment
No sample size re-estimation will be performed.
9.3.
Data Analysis Considerations
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
 
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sampling times recorded during the study.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-
life (t1/2).  
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model.
9.3.1.
Analysis Populations
9.3.1.1.
Safety Population
All subjects enrolled into the study who have received at least one dose of investigational 
product will be included in the Safety Population.
9.3.1.2.
Pharmacokinetic Concentration Population
The PK Concentration Population will include all subjects for whom at least one PK 
sample was obtained and analyzed.
9.3.1.3.
Pharmacokinetic Parameter Population
For each PK parameter, the PK Parameter Population will include all subjects who 
provide PK parameter data.
9.3.2.
Interim Analysis
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in order to direct development of fixed dose formulations for subsequent 
study parts and the planned BE study.
9.4.
Key Elements of Analysis Plan
9.4.1.
Pharmacokinetic Analyses
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
 
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treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D. 
Following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using a mixed effects model with fixed effect 
terms for period and treatment. Subject will be treated as a random effect in the model. 
Point estimates and their associated 90% confidence intervals will be constructed for the 
differences, F1 – R, F2 – R, F3 – R, F4 – R for Part 1 and similarly for Part 2 (as needed); 
and X1-X2, Y1-Y2 for Part 3. The point estimates and their associated 90% confidence 
intervals will then be back-transformed to provide point estimates and 90% confidence 
intervals for the ratios, F1:R, F2-R:F3-R, F4-R; and X1:X2, Y1:Y2.
Tmax of FDC formulations will be analyzed with the non-parametric Wilcoxon Matched 
Pairs Method to compute point estimate and associated 90% confidence intervals for the 
median difference, F1 R, F2 R, F3 R, F4 R and X1:X2, Y1:Y2.
9.4.2.
Safety Analyses
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
10.
STUDY GOVERNANCE CONSIDERATIONS
10.1.
Posting of Information on Publicly Available Clinical Trial 
Registers
Study information from this protocol will be posted on publicly available clinical trial 
registers before enrollment of subjects begins.
10.2.
Regulatory and Ethical Considerations, Including the 
Informed Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.
The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent

Investigator reporting requirements (e.g. reporting of AEs/SAEs/protocol 
deviations to IRB/IEC)
 
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
GSK will provide full details of the above procedures, either verbally, in writing, 
or both.

Signed informed consent must be obtained for each subject prior to participation 
in the study

The IEC/IRB, and where applicable the regulatory authority, approve the clinical 
protocol and all optional assessments, including genetic research.  

Optional assessments (including those in a separate protocol and/or under separate 
informed consent) and the clinical protocol should be concurrently submitted for 
approval unless regulation requires separate submission.  

Approval of the optional assessments may occur after approval is granted for the 
clinical protocol where required by regulatory authorities.  In this situation, 
written approval of the clinical protocol should state that approval of optional 
assessments is being deferred and the study, with the exception of the optional 
assessments, can be initiated.
10.3.
Quality Control (Study Monitoring)

In accordance with applicable regulations including GCP, and GSK procedures, 
GSK monitors will contact the site prior to the start of the study to review with the 
site staff the protocol, study requirements, and their responsibilities to satisfy 
regulatory, ethical, and GSK requirements.  

When reviewing data collection procedures, the discussion will also include 
identification, agreement and documentation of data items for which the CRF will 
serve as the source document.
GSK will monitor the study and site activity to verify that the:

Data are authentic, accurate, and complete.

Safety and rights of subjects are being protected.

Study is conducted in accordance with the currently approved protocol and any 
other study agreements, GCP, and all applicable regulatory requirements.
The investigator and the head of the medical institution (where applicable) agrees to 
allow the monitor direct access to all relevant documents
10.4.
Quality Assurance

To ensure compliance with GCP and all applicable regulatory requirements, GSK 
may conduct a quality assurance assessment and/or audit of the site records, and 
the regulatory agencies may conduct a regulatory inspection at any time during or 
after completion of the study. 

In the event of an assessment, audit or inspection, the investigator (and institution) 
must agree to grant the advisor(s), auditor(s) and inspector(s) direct access to all 
relevant documents and to allocate their time and the time of their staff to discuss 
the conduct of the study, any findings/relevant issues and to implement any 
corrective and/or preventative actions to address any findings/issues identified.
 
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10.5.
Study and Site Closure

Upon completion or premature discontinuation of the study, the GSK monitor will 
conduct site closure activities with the investigator or site staff, as appropriate, in 
accordance with applicable regulations including GCP, and GSK Standard 
Operating Procedures.

GSK reserves the right to temporarily suspend or prematurely discontinue this 
study at any time for reasons including, but not limited to, safety or ethical issues 
or severe non-compliance. For multicenter studies, this can occur at one or more 
or at all sites.  

If GSK determines such action is needed, GSK will discuss the reasons for taking 
such action with the investigator or the head of the medical institution (where 
applicable).  When feasible, GSK will provide advance notification to the 
investigator or the head of the medical institution, where applicable, of the 
impending action.

If the study is suspended or prematurely discontinued for safety reasons, GSK will 
promptly inform all investigators, heads of the medical institutions (where 
applicable) and/or institution(s) conducting the study. GSK will also promptly 
inform the relevant regulatory authorities of the suspension or premature 
discontinuation of the study and the reason(s) for the action.  

If required by applicable regulations, the investigator or the head of the medical 
institution (where applicable) must inform the IRB/IEC promptly and provide the 
reason for the suspension or premature discontinuation.
10.6.
Records Retention

Following closure of the study, the investigator or the head of the medical 
institution (where applicable) must maintain all site study records (except for 
those required by local regulations to be maintained elsewhere), in a safe and 
secure location.  

The records must be maintained to allow easy and timely retrieval, when needed 
(e.g., for a GSK audit or regulatory inspection) and must be available for review 
in conjunction with assessment of the facility, supporting systems, and relevant 
site staff.  

Where permitted by local laws/regulations or institutional policy, some or all of 
these records can be maintained in a format other than hard copy (e.g., microfiche, 
scanned, electronic); however, caution needs to be exercised before such action is 
taken.  

The investigator must ensure that all reproductions are legible and are a true and 
accurate copy of the original and meet accessibility and retrieval standards, 
including re-generating a hard copy, if required.  Furthermore, the investigator 
must ensure there is an acceptable back-up of these reproductions and that an 
acceptable quality control process exists for making these reproductions.
 
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
GSK will inform the investigator of the time period for retaining these records to 
comply with all applicable regulatory requirements.  The minimum retention time 
will meet the strictest standard applicable to that site for the study, as dictated by 
any institutional requirements or local laws or regulations, GSK 
standards/procedures, and/or institutional requirements.  

The investigator must notify GSK of any changes in the archival arrangements, 
including, but not limited to, archival at an off-site facility or transfer of 
ownership of the records in the event the investigator is no longer associated with 
the site.
10.7.
Provision of Study Results to Investigators, Posting of 
Information on Publically Available Clinical Trials Registers 
and Publication
Where required by applicable regulatory requirements, an investigator signatory will be 
identified for the approval of the clinical study report.  The investigator will be provided 
reasonable access to statistical tables, figures, and relevant reports and will have the 
opportunity to review the complete study results at a GSK site or other mutually-
agreeable location.
GSK will also provide the investigator with the full summary of the study results.  The 
investigator is encouraged to share the summary results with the study subjects, as 
appropriate.
GSK will provide the investigator with the randomization codes for their site only after 
completion of the full statistical analysis.
The procedures and timing for public disclosure of the results summary and for 
development of a manuscript for publication will be in accordance with GSK Policy.
A manuscript will be progressed for publication in the scientific literature if the results 
provide important scientific or medical knowledge.
 
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11.
REFERENCES
Adcirca: EMA Summary of product Characteristics, Eli Lily, last updated 29-Sep-2015 at 
time of publication
EMEA guidelines on clinical development of fixed dose combination medicinal products. 
CHMP/EWP/240/95 Rev.1. 2008
Forgue S, Patterson B, Bedding A, et al. Tadalafil pharmacokinetics in healthy subjects. 
British Journal of Clinical Pharmacology. 20-Dec-2005
Galiè N et al. Initial Use of Ambrisentan plus Tadalafil in Pulmonary Arterial 
Hypertension. N Engl J Med 2015a;373:834-44.
Galiè N, Humbert M, Vachiery J.L et al. 2015 ESC/ERS Guidelines for the diagnosis and 
treatment of pulmonary hypertension. European Heart Journal Advance Access, 
published August 29, 2015b
Ghofrani AH, Galiè N, Grimminger F, et al. N Engl J Med 2013;369:330-40. Riociguat 
for the Treatment of Pulmonary Arterial Hypertension
Gilead Clinical Study report GS-US-300-0112: A Phase 1, Open-Label, Randomized, 
Two-Period, Crossover Study to Assess the Pharmacokinetics of Ambrisentan Alone and 
in the Presence of Multiple Doses of Tadalafil, and to assess the Pharmacokinetics of 
Tadalafil Alone and in the Presence of Multiple Doses of Ambrisentan in Healthy Adult 
Volunteers. Version 1.0, dated 27-June-2008
Gilead Investigator Brochure, Letaris, Volibris, Fifteenth edition: 06 April 2015
GSK Clinical Study Report for study AMB112565: AMBITION: A Randomized, 
Multicenter Study of First-Line Ambrisentan and Tadalafil Combination Therapy in 
Subjects with Pulmonary Arterial Hypertension. GSK document 2014N193963_00, dated 
13-Nov-14
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(a): 24 Table 3-2
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(b): 28.
Liang F, Yang S, Yao L, Belardinelli L, Shryock J. Hypertension. 2012 Mar;59(3):705-
11. Ambrisentan and tadalafil synergistically relax endothelin-induced contraction of rat 
pulmonary arteries.
Pulido T, Adzerikho I, Channick RN, et al. N Engl J Med. 2013 Aug 29;369(9):809-18. 
Macitentan and Morbidity and Mortality in Pulmonary Arterial Hypertension
 
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Spence R, Mandagere A, Harrison B, et al.  J Pharm Sci. 2009 Dec;98(12):4962-74. No 
clinically relevant pharmacokinetic and safety interactions of ambrisentan in combination 
with tadalafil in healthy volunteers. 
Volibris: EMA Summary of Product Characteristics, GSK, variation II/41, at time of 
publication, 2015.
 
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12.
APPENDICES
12.1.
Appendix 1: Abbreviations and Trademarks
Abbreviations
AE
Adverse Event
API
Active Pharmaceutical Ingredient
AUC
Area Under Curve
BE
Bioequivalence
CDISC
Clinical Data Interchange Standards Consortium
cGMP
Cyclic guanosine monophosphate
Cmax
Maximum concentration
CI
Confidence Interval
CV
Coefficient of variance 
CVw
Coefficient of variance within subject
EMA
European Medicines Agency 
ERA
Endothelin Receptor Antagonists
ERS
European Respiratory Society 
ESC
European Society of Cardiology
ET-1
Endothelin Receptor - 1
EU
European Union
FDA
Federal Drug Agency
FDC
Fixed Dose Combination
NO
Nitrous Oxide
PDE-5
Phosphodiesterase type 5
SAE
Serious Adverse Events
SRM
Study Reference Manual
tmax
Time to maximum concentration
t1/2
Half Life
WHO
World Health Organisation
Trademark Information
Trademarks of the GlaxoSmithKline 
group of companies
Trademarks not owned by the 
GlaxoSmithKline group of companies
NONE
Adcirca
WinNonlin
 
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12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments
Phase I Liver chemistry stopping criteria have been designed to assure subject safety and 
to evaluate liver event aetiology (in alignment with the FDA premarketing clinical liver 
safety guidance).
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf.
Phase I liver chemistry stopping criteria and required follow up assessments
Liver Chemistry Stopping Criteria – Liver Stopping Event
ALT-absolute
ALT≥3xULN
If  ALT≥3xULN  AND bilirubin1,2 2xULN (>35% direct bilirubin) or  INR >1.5, 
Report as an SAE.
See additional Actions and Follow-Up Assessments listed below
Required Actions and Follow up Assessments following Liver Stopping Event
Actions
Follow-Up Assessments

Immediately discontinue study treatment 

Report the event to GSK within 24 hours

Complete the liver event CRF, and complete 
an SAE data collection tool if the event also 
meets the criteria for an SAE2

Perform liver event follow up assessments 

Monitor the subject until liver chemistries 
resolve, stabilise, or return to within baseline 
(see MONITORING below)
MONITORING:
If ALT≥3xULN AND bilirubin 2xULN  or INR 
>1.5

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within  24 
hrs

Monitor subjects twice weekly until liver 
chemistries resolve, stabilise or return to 
within baseline

Viral hepatitis serology3

Blood sample for pharmacokinetic (PK) 
analysis, obtained within 96hrs of last dose

Serum creatine phosphokinase (CPK) and 
lactate dehydrogenase (LDH).

Fractionate bilirubin, if total 
bilirubin2xULN

Obtain complete blood count with 
differential to assess eosinophilia

Record the appearance or worsening of 
clinical symptoms of liver injury, or 
hypersensitivity, on the AE report form

Record use of concomitant medications on 
the concomitant medications report form 
including acetaminophen, herbal remedies, 
other over the counter medications.

Record alcohol use on the liver event 
alcohol intake case report form
If ALT≥3xULN AND bilirubin 2xULN  or 
 
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Liver Chemistry Stopping Criteria – Liver Stopping Event

A specialist or hepatology consultation is 
recommended
If ALT≥3xULN AND bilirubin < 2xULN  and 
INR ≤1.5:

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within 24-
72 hrs

Monitor subjects weekly until liver 
chemistries resolve, stabilize or return to 
within baseline
INR >1.5:

Anti-nuclear antibody, anti-smooth muscle 
antibody, Type 1 anti-liver kidney 
microsomal antibodies, and quantitative 
total immunoglobulin G (IgG or gamma 
globulins).

Serum acetaminophen adduct HPLC assay 
(quantifies potential acetaminophen 
contribution to liver injury in subjects with 
definite or likely acetaminophen use in the 
preceding week [James, 2009].

Liver imaging (ultrasound, magnetic 
resonance, or computerised tomography) 
and /or liver biopsy to evaluate liver 
disease; complete Liver Imaging and/or 
Liver Biopsy CRF forms.
1.
Serum bilirubin fractionation should be performed if testing is available. If serum bilirubin fractionation is not 
immediately available, discontinue study treatment for that subject if ALT 3xULN and bilirubin 2xULN. 
Additionally, if serum bilirubin fractionation testing is unavailable, record presence of detectable urinary bilirubin on 
dipstick, indicating direct bilirubin elevations and suggesting liver injury. 
2.
All events of ALT 3xULN and bilirubin 2xULN (>35% direct bilirubin) or ALT 3xULN and INR>1.5, if INR 
measured, which may indicate severe liver injury (possible ‘Hy’s Law’), must be reported as an SAE (excluding 
studies of hepatic impairment or cirrhosis); INR measurement is not required and the threshold value stated will 
not apply to subjects receiving anticoagulants
3.
Includes: Hepatitis A IgM antibody; Hepatitis B surface antigen and Hepatitis B Core Antibody (IgM); Hepatitis C 
RNA; Cytomegalovirus IgM antibody;  Epstein-Barr viral capsid antigen IgM antibody (or if unavailable, obtain 
heterophile antibody or monospot testing);  Hepatitis E IgM antibody 
4.
PK sample may not be required for subjects known to be receiving placebo or non-GSK comparator treatments.  
Record the date/time of the PK blood sample draw and the date/time of the last dose of study treatment prior to 
blood sample draw on the CRF. If the date or time of the last dose is unclear, provide the subject’s best 
approximation. If the date/time of the last dose cannot be approximated OR a PK sample cannot be collected in 
the time period indicated above, do not obtain a PK sample. Instructions for sample handling and shipping are in 
the SRM. 
References
James LP, Letzig L, Simpson PM, Capparelli E, Roberts DW, Hinson JA, Davern TJ, Lee 
WM. Pharmacokinetics of Acetaminophen-Adduct in Adults with Acetaminophen 
Overdose and Acute Liver Failure. Drug Metab Dispos 2009; 37:1779-1784.
 
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12.3.
Appendix 3: Definition of and Procedures for Recording, 
Evaluating, Follow-Up and Reporting of Adverse Events
12.3.1.
Definition of Adverse Events
Adverse Event Definition:

An AE is any untoward medical occurrence in a patient or clinical investigation 
subject, temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product.

NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal product.
Events meeting AE definition include:

Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (e.g., ECGs, radiological scans, vital signs 
measurements), including those that worsen from baseline, and felt to be clinically 
significant in the medical and scientific judgement of the investigator.

Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

New conditions detected or diagnosed after study treatment administration even 
though it may have been present prior to the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected interaction.

Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication (overdose per se will not be reported as an 
AE/SAE unless this is an intentional overdose taken with possible suicidal/self-
harming intent.  This should be reported regardless of sequelae).
Events NOT meeting definition of an AE include:

Any clinically significant abnormal laboratory findings or other abnormal safety 
assessments which are associated with the underlying disease, unless judged by the 
investigator to be more severe than expected for the subject’s condition.

The disease/disorder being studied or expected progression, signs, or symptoms of 
the disease/disorder being studied, unless more severe than expected for the subject’s 
condition.

Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that 
leads to the procedure is an AE.

Situations where an untoward medical occurrence did not occur (social and/or 
 
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convenience admission to a hospital).

Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen.
12.3.2.
Definition of Serious Adverse Events
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, 
death due to progression of disease, etc).
Serious Adverse Event (SAE) is defined as any untoward medical occurrence that, 
at any dose:
a.
Results in death
b.
Is life-threatening
NOTE:
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
subject was at risk of death at the time of the event.  It does not refer to an event, which 
hypothetically might have caused death, if it were more severe.
c.
Requires hospitalization or prolongation of existing hospitalization
NOTE:

In general, hospitalization signifies that the subject has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for 
observation and/or treatment that would not have been appropriate in the physician’s 
office or out-patient setting.  Complications that occur during hospitalization are 
AEs.  If a complication prolongs hospitalization or fulfills any other serious criteria, 
the event is serious.  When in doubt as to whether “hospitalization” occurred or was 
necessary, the AE should be considered serious.

Hospitalization for elective treatment of a pre-existing condition that did not worsen 
from baseline is not considered an AE.
d.
Results in disability/incapacity
NOTE:

The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.

This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, 
and accidental trauma (e.g. sprained ankle) which may interfere or prevent everyday 
life functions but do not constitute a substantial disruption
 
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e.
Is a congenital anomaly/birth defect
f.
Other situations:

Medical or scientific judgment should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be 
immediately life-threatening or result in death or hospitalization but may jeopardize 
the subject or may require medical or surgical intervention to prevent one of the 
other outcomes listed in the above definition.  These should also be considered 
serious.

Examples of such events are invasive or malignant cancers, intensive treatment in an 
emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse
g.
Is associated with liver injury and impaired liver function defined as:

ALT 3xULN and total bilirubin* 2xULN (>35% direct), or

ALT 3xULN and INR** > 1.5.
* Serum bilirubin fractionation should be performed if testing is available; if unavailable, 
measure urinary bilirubin via dipstick.  If fractionation is unavailable and ALT 3xULN 
and total bilirubin 2xULN, then the event is still to be reported as an SAE.
** INR testing not required per protocol and the threshold value does not apply to 
subjects receiving anticoagulants.  If INR measurement is obtained, the value is to be 
recorded on the SAE form.

Refer to Section 12.2 for the required liver chemistry follow up instructions
12.3.3.
Definition of Cardiovascular Events
Cardiovascular Events (CV) Definition:
Investigators will be required to fill out the specific CV event page of the CRF for the 
following AEs and SAEs:

Myocardial infarction/unstable angina

Congestive heart failure

Arrhythmias

Valvulopathy

Pulmonary hypertension

Cerebrovascular events/stroke and transient ischemic attack

Peripheral arterial thromboembolism

Deep venous thrombosis/pulmonary embolism
 
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
Revascularization
12.3.4.
Recording of AEs and SAEs
AEs and SAE Recording:

When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) 
relative to the event.

The investigator will then record all relevant information regarding an AE/SAE in 
the CRF

It is not acceptable for the investigator to send photocopies of the subject’s medical 
records to GSK in lieu of completion of the GSK, AE/SAE CRF page.

There may be instances when copies of medical records for certain cases are 
requested by GSK.  In this instance, all subject identifiers, with the exception of the 
subject number, will be blinded on the copies of the medical records prior to 
submission of to GSK.

The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information.  In such cases, the diagnosis will be 
documented as the AE/SAE and not the individual signs/symptoms.

Subject-completed Value Evidence and Outcomes questionnaires and the collection 
of AE data are independent components of the study.

Responses to each question in the Value Evidence and Outcomes questionnaire will 
be treated in accordance with standard scoring and statistical procedures detailed by 
the scale’s developer.

The use of a single question from a multidimensional health survey to designate a 
cause-effect relationship to an AE is inappropriate.
12.3.5.
Evaluating AEs and SAEs
Assessment of Intensity
The investigator will make an assessment of intensity for each AE and SAE reported 
during the study and will assign it to one of the following categories:

Mild:  An event that is easily tolerated by the subject, causing minimal discomfort 
and not interfering with everyday activities.

Moderate:  An event that is sufficiently discomforting to interfere with normal 
everyday activities

Severe:  An event that prevents normal everyday activities. - an AE that is assessed 
as severe will not be confused with an SAE. Severity is a category utilized for rating 
 
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the intensity of an event; and both AEs and SAEs can be assessed as severe.

An event is defined as ‘serious’ when it meets at least one of the pre-defined 
outcomes as described in the definition of an SAE.
Assessment of Causality

The investigator is obligated to assess the relationship between study treatment and 
the occurrence of each AE/SAE.

A "reasonable possibility" is meant to convey that there are facts/evidence or 
arguments to suggest a causal relationship, rather than a relationship cannot be ruled 
out.

The investigator will use clinical judgment to determine the relationship.

Alternative causes, such as natural history of the underlying diseases, concomitant 
therapy, other risk factors, and the temporal relationship of the event to the study 
treatment will be considered and investigated.

The investigator will also consult the Investigator Brochure (IB) and/or Product 
Information, for marketed products, in the determination of his/her assessment.

For each AE/SAE the investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality.

There may be situations when an SAE has occurred and the investigator has minimal 
information to include in the initial report to GSK.  However, it is very important 
that the investigator always make an assessment of causality for every event 
prior to the initial transmission of the SAE data to GSK.

The investigator may change his/her opinion of causality in light of follow up 
information, amending the SAE data collection tool accordingly.

The causality assessment is one of the criteria used when determining regulatory 
reporting requirements.
Follow up of AEs and SAEs

The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as may be indicated or as requested by GSK to 
elucidate as fully as possible the nature and/or causality of the AE or SAE.

The investigator is obligated to assist.  This may include additional laboratory tests 
or investigations, histopathological examinations or consultation with other health 
care professionals.

If a subject dies during participation in the study or during a recognized follow up 
period, the investigator will provide GSK with a copy of any post-mortem findings, 
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
New or updated information will be recorded in the originally completed CRF.

The investigator will submit any updated SAE data to GSK within the designated 
reporting time frames.
12.3.6.
Reporting of SAEs to GSK
SAE reporting to GSK via paper CRF

Facsimile transmission of the SAE paper CRF is the preferred method to transmit 
this information to the Medical Monitor or the SAE coordinator

In rare circumstances and in the absence of facsimile equipment, notification by 
telephone is acceptable, with a copy of the SAE data collection tool sent by 
overnight mail

Initial notification via the telephone does not replace the need for the investigator to 
complete and sign the SAE CRF pages within the designated reporting time frames.

Contacts for SAE receipt can be found at this beginning of the protocol on the 
Sponsor/Medical Monitor Contact Information page.
 
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12.4.
Appendix 4: High Fat Meal Content

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
The standard high-fat meal will be the meal suggested by the US FDA in their 2002 draft 
guidance on food-effect bioavailability and bioequivalence studies. Approximately 50% 
of the caloric content of the meal is from fat and the meal is high in calories 
(approximately 1000 calories). Substitutions in this test meal can be made as long as the 
meal provides a similar amount of calories from protein, carbohydrate, and fat and has 
comparable meal volume and viscosity.
US Department of Health and Human Services.  Guidance for Industry. Food-Effect 
Bioavailability and Fed Bioequivalence Studies.  Guidance. Center for Drug Evaluation 
and Research (CDER). Food and Drug Administration; 2002.
 
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12.5.
Appendix 5: Protocol Changes
Amendment 01
The purpose of this amendment is to clarify the hypotension withdrawal criteria in 
section 5.4.3, address changes to GSK medical monitor and minor administrative 
changes.
Summary of Changes
Section 1 PROTOCOL SYNOPSIS FOR STUDY 201964
AND
Section 3 OBJECTIVES AND ENDPOINTS
CHANGE FROM
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assed by vital signs, ECG, clinical laboratory 
safety data and review of adverse events
CHANGE TO
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assessed by vital signs, ECG, clinical 
laboratory safety data and review of adverse 
events
 
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Section 5.4.3 Hypotension 
CHANGE FROM
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 or diastolic <50 confirmed by triplicate reading 
taken up to 5 minutes apart.
CHANGE TO
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 mm HG and diastolic <50 mm HG confirmed 
by triplicate reading taken up to 5 minutes apart and is judged clinically 
significant and symptomatic by the investigator. 
Section 6.3 Treatment Assignment
ADDITION 
Table 6 Treatment Sequence assignments for Part 1, Part 2 and Part 3
Part 2 and 
Part 3 
merged 
together
5 (Two FDCs selected 
from Part 1)
X1 X2 R Y1 Y2
X2 Y1 X1 Y2 R
Y1 Y2 X2 R X1
Y2 R Y1 X1 X2
R X1 Y2 X2 Y1
Y2 Y1 R X2 X1
R Y2 X1 Y1 X2
X1 R X2 Y2 Y1
X2 X1 Y1 R Y2
Y1 X2 Y2 X1 R
1:1:1:1:1:1:1:1:1:1
3 (One FDC selected 
from Part 1)
X1 X2 R
R X2 X1
X2 R X1
F1 R X2
R X1 X2
X2 X1 R
3:3:3:3:3:3 + 2 
random
 
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201964
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TITLE PAGE
Division:  Worldwide Development
Information Type: Clinical Protocol
Title:
A Phase 1 study to demonstrate the relative bioavailability of 
fixed dose combinations of ambrisentan and tadalafil in healthy 
subjects
Compound Number:
GSK3380154, GSK1325760 and GF196960
Development Phase
I
Effective Date:
16-DEC-2015
Author(s):
 
Copyright 2015 the GlaxoSmithKline group of companies.  All rights reserved.  
Unauthorised copying or use of this information is prohibited.
 
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MEDICAL MONITOR/SPONSOR INFORMATION PAGE
Medical Monitor/SAE Contact Information:
Role
Name
Day Time Phone Number and 
email address
After-hours 
Phone/Cell/
Pager Number
Fax Number
Primary Medical 
Monitor 
Site to scan 
and send to 
email
Secondary 
Medical Monitor
Site to scan 
and send to 
email
SAE contact  
information 
Site to scan 
and send to 
email
Sponsor Legal Registered Address:
GlaxoSmithKline Research & Development Limited
980 Great West Road
Brentford
Middlesex, TW8 9GS
UK
In some countries, the clinical trial sponsor may be the local GlaxoSmithKline Affiliate 
Company (or designee). If applicable, the details of the alternative Sponsor and contact 
person in the territory will be provided to the relevant regulatory authority as part of the 
clinical trial application.   
Regulatory Agency Identifying Number(s): EudraCT number 2015-004140-18
 
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INVESTIGATOR PROTOCOL AGREEMENT PAGE

I confirm agreement to conduct the study in compliance with the protocol.

I acknowledge that I am responsible for overall study conduct. I agree to 
personally conduct or supervise the described study.

I agree to ensure that all associates, colleagues and employees assisting in the 
conduct of the study are informed about their obligations. Mechanisms are in 
place to ensure that site staff receives the appropriate information throughout the 
study.
Investigator Name:  ______________________________
Investigator Signature
Date
 
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TABLE OF CONTENTS
PAGE
1.
PROTOCOL SYNOPSIS FOR STUDY 201964........................................................8
2.
INTRODUCTION....................................................................................................11
2.1.
Study Rationale ..........................................................................................11
2.2.
Brief Background ........................................................................................11
3.
OBJECTIVE(S) AND ENDPOINT(S) ......................................................................14
4.
STUDY DESIGN ....................................................................................................14
4.1.
Overall Design ............................................................................................14
4.2.
Treatment and Duration..............................................................................16
4.3.
Type and Number of Subjects.....................................................................19
4.4.
Design Justification.....................................................................................20
4.5.
Dose Justification........................................................................................20
4.6.
Benefit: Risk Assessment ...........................................................................20
4.6.1.
Risk Assessment .........................................................................21
4.6.2.
Benefit Assessment.....................................................................23
4.6.3.
Overall Benefit: Risk Conclusion..................................................23
5.
SELECTION OF STUDY POPULATION AND WITHDRAWAL CRITERIA .............23
5.1.
Inclusion Criteria.........................................................................................24
5.2.
Exclusion Criteria........................................................................................25
5.3.
Screening/Baseline/Run-in Failures............................................................27
5.4.
Withdrawal/Stopping Criteria.......................................................................27
5.4.1.
Liver Chemistry Stopping Criteria ................................................27
5.4.1.1.
Study Treatment Restart or Rechallenge....................28
5.4.2.
QTc Stopping Criteria ..................................................................28
5.4.3.
Hypotension.................................................................................29
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria .........................29
5.5.
Subject and Study Completion....................................................................29
6.
STUDY TREATMENT ............................................................................................29
6.1.
Investigational Product and Other Study Treatment....................................29
6.2.
Medical Devices..........................................................................................31
6.3.
Treatment Assignment................................................................................31
6.4.
Blinding.......................................................................................................33
6.5.
Packaging and Labeling..............................................................................33
6.6.
Preparation/Handling/Storage/Accountability..............................................33
6.7.
Compliance with Study Treatment Administration.......................................33
6.8.
Treatment of Study Treatment Overdose....................................................33
6.9.
Treatment after the End of the Study ..........................................................34
6.10.
Lifestyle and/or Dietary Restrictions............................................................34
6.10.1.
Meals and Dietary Restrictions ....................................................34
6.10.2.
Caffeine, Alcohol, and Tobacco ...................................................35
6.10.3.
Activity .........................................................................................35
6.11.
Concomitant Medications and Non-Drug Therapies....................................35
6.11.1.
Permitted Medications and Non-Drug Therapies..........................35
6.11.2.
Prohibited Medications and Non-Drug Therapies.........................35
 
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7.
STUDY ASSESSMENTS AND PROCEDURES .....................................................35
7.1.
Time and Events Table...............................................................................37
7.2.
Screening and Critical Baseline Assessments ............................................38
7.2.1.
Demographic/Medical History Assessments ................................38
7.3.
Pharmacokinetics .......................................................................................38
7.3.1.
Blood Sample Collection..............................................................38
7.3.2.
Sample Analysis ..........................................................................38
7.4.
Safety .........................................................................................................39
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)..........39
7.4.1.1.
Time period and Frequency for collecting AE 
and SAE information...................................................39
7.4.1.2.
Method of Detecting AEs and SAEs ...........................40
7.4.1.3.
Follow-up of AEs and SAEs........................................40
7.4.1.4.
Regulatory Reporting Requirements for SAEs............40
7.4.2.
Physical Exams ...........................................................................40
7.4.3.
Vital Signs....................................................................................41
7.4.4.
Electrocardiogram (ECG).............................................................41
7.4.5.
Clinical Safety Laboratory Assessments ......................................41
8.
DATA MANAGEMENT ...........................................................................................43
9.
STATISTICAL CONSIDERATIONS AND DATA ANALYSES .................................43
9.1.
Hypotheses.................................................................................................43
9.2.
Sample Size Considerations.......................................................................43
9.2.1.
Sample Size Assumptions ...........................................................43
9.2.2.
Sample Size Sensitivity................................................................44
9.2.3.
Sample Size Re-estimation or Adjustment...................................44
9.3.
Data Analysis Considerations .....................................................................44
9.3.1.
Analysis Populations....................................................................45
9.3.1.1.
Safety Population .......................................................45
9.3.1.2.
Pharmacokinetic Concentration Population ................45
9.3.1.3.
Pharmacokinetic Parameter Population......................45
9.3.2.
Interim Analysis ...........................................................................45
9.4.
Key Elements of Analysis Plan ...................................................................45
9.4.1.
Pharmacokinetic Analyses...........................................................45
9.4.2.
Safety Analyses...........................................................................46
10. STUDY GOVERNANCE CONSIDERATIONS........................................................46
10.1.
Posting of Information on Publicly Available Clinical Trial Registers............46
10.2.
Regulatory and Ethical Considerations, Including the Informed 
Consent Process ........................................................................................46
10.3.
Quality Control (Study Monitoring) ..............................................................47
10.4.
Quality Assurance.......................................................................................47
10.5.
Study and Site Closure ...............................................................................48
10.6.
Records Retention......................................................................................48
10.7.
Provision of Study Results to Investigators, Posting of Information 
on Publically Available Clinical Trials Registers and Publication .................49
11. REFERENCES.......................................................................................................50
12. APPENDICES........................................................................................................52
12.1.
Appendix 1: Abbreviations and Trademarks................................................52
 
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12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up 
Assessments ..............................................................................................53
12.3.
Appendix 3: Definition of and Procedures for Recording, Evaluating, 
Follow-Up and Reporting of Adverse Events...............................................55
12.3.1.
Definition of Adverse Events........................................................55
12.3.2.
Definition of Serious Adverse Events...........................................56
12.3.3.
Definition of Cardiovascular Events .............................................57
12.3.4.
Recording of AEs and SAEs ........................................................58
12.3.5.
Evaluating AEs and SAEs............................................................58
12.3.6.
Reporting of SAEs to GSK...........................................................60
12.4.
Appendix 4: High Fat Meal Content ............................................................61
 
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1.
PROTOCOL SYNOPSIS FOR STUDY 201964
Rationale
This study is designed to select one, or more fixed dose combinations (FDCs) of 
ambrisentan and tadalafil for further development and to provide pharmacokinetic data to 
enable a pivotal bioequivalence study (BE). Dependent on formulation work, the study 
will allow up to 8 new FDCs to be compared with the reference of ambrisentan and 
tadalafil monotherapies taken concurrently. The study would also allow for up to 2 of the 
new formulations, that may be taken in to a BE study, to be tested for any effect on 
pharmacokinetics of the FDC in both fed and fasted state.
Objective(s)/Endpoint(s)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following administration 
of candidate tablet formulations of the FDC 
(ambrisentan 10 mg + tadalafil 40 mg) relative to 
reference monotherapies tested (ambrisentan 10 
mg & tadalafil 40 mg) taken concurrently in healthy 
human subjects under fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), 
and AUC(0-t) of ambrisentan and tadalafil in 
FDC and reference treatments
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2 of 
ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assed by vital signs, ECG, clinical laboratory 
safety data and review of adverse events
Overall Design
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects.
All subjects will attend the unit for Screening within 31 days of their first dose. Eligible 
subjects will be randomised to order of treatment in that study parts in which they are 
included and the order in which these treatments are administered will be in accordance 
with the randomisation schedule.
 
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There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose. The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil 
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part. 
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and  be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast.
Treatment Arms and Duration
The proposed treatment arms for each study part are described here; however the 
treatments in Parts 2 and 3 may be changed dependent on the utility and results from the 
previous part.
 
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Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil 
FDC1 (10mg/40mg)
ambrisentan and tadalafil 
FDC5 (10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil 
FDC2 (10mg/40mg)
ambrisentan and tadalafil 
FDC6 (10mg/40mg)
ambrisentan and tadalafil 
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil 
FDC3 (10mg/40mg)
ambrisentan and tadalafil 
FDC7 (10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil
FDC4 (10mg/40mg)
ambrisentan and tadalafil 
FDC8 (10mg/40mg)
ambrisentan and tadalafil 
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil 
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less FDC treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. The minimum will be approximately 3 weeks and the maximum will be 
approximately 18 weeks. 
Type and Number of Subjects
A maximum of 20 healthy adult subjects will be randomized, to each study part, such that 
approximately 16 evaluable subjects complete each part of the study.
Analysis
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and Part 2, (ii) estimate the bioavailability of the formulation(s) of 
the FDC formulations, taken in Part 2, if used for food effect and  Part 3, in the fed state 
relative to the fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
 
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2.
INTRODUCTION
Ambrisentan (E.U. trade name: Volibris), an orally active endothelin receptor antagonists 
(ERA) that is selective for ETA. Once daily dosing at 5 or 10 mg, was first approved on 
15 June 2007 in the US and on 21 April 2008 in the European Union (EU) and is 
currently approved in over 50 countries. In the EU, ambrisentan is indicated for treatment 
of pulmonary arterial hypertension (PAH) in adult patients of WHO Functional Class 
(FC) II to III, including use in combination treatment (Volibris, EMA SmPC, Section 
5.1). Efficacy has been shown in idiopathic PAH (IPAH) and in PAH associated with 
connective tissue disease
Tadalafil (E.U. trade name: Adcirca) is an orally active selective inhibitor of the enzyme 
PDE-5, the primary cGMP-hydrolyzing enzyme in smooth muscle. In the EU, Adcirca is 
indicated in adults for the treatment of PAH classified as WHO FC II and III, to improve 
exercise capacity (Adcirca, EMA, SmPC).
A recently completed study (Galiè N, 2015) has show that patients with PAH who started 
initial combination therapy with ambrisentan and tadalafil had a significantly lower risk 
of clinical-failure events compared to those that started with ambrisentan or tadalafil 
monotherapy. Ambrisentan has recently received EU approval (20 November 2015) for 
use in combination treatment with tadalafil (Volibris, EMA SmPC, Section 5.1). 
This pilot study will investigate the relative bioavailability of new fixed dose 
combinations (FDC) of ambrisentan and tadalafil, compared to the two monotherapies 
taken concurrently in healthy subjects
2.1.
Study Rationale
This study is designed to select one, or more, FDCs of ambrisentan and tadalafil for 
further development and to provide pharmacokinetic data to enable a pivotal 
bioequivalence study (BE). Dependent on formulation work, the study will allow up to 8 
new FDCs to be compared with the reference of ambrisentan and tadalafil monotherapies 
taken concurrently. The study would also allow for up to 2 of the new formulations, that 
may be taken in to a BE study, to be tested for any effect on pharmacokinetics of the 
FDC in both fed and fasted state.
The formulation(s) to be taken forward into the BE study will be primarily chosen on the 
test/reference ratio for both AUC and Cmax for both components ideally the 90% 
confidence interval (CI) for the ratio would fall in the range of 0.80 and 1.25, with 
successful formulations having test/reference ratios close to unity and with low intra 
subject variability indicated by a narrow CI. If a number of candidate formulations 
successfully meet these criteria then other factors, including, between subject variability,
tablet size, cost, ease of manufacture and stability would be considered. 
2.2.
Brief Background
Pulmonary Arterial Hypertension (PAH) is a progressive, life threatening disease that, 
despite the emergence of new treatments, still has a poor long term prognosis (akin to 
many cancers). Treatments currently approved for the treatment of PAH target 3 
 
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biological pathways, namely; endothelin (ET-1), nitric oxide (NO) and prostacyclin 
pathways. Due to the severity and progressive nature of the disease, combination therapy 
with agents targeting these different pathways has become increasingly utilised over the 
years. The evidence for sequential combination treatment has grown and it is now 
recommended in the latest treatment guidelines (Galie`, 2015) and the recent EU 
approval of Ambrisentan for combination treatment of Ambrisentan plus Tadalafil for 
PAH. In practice the combined use of medications targeting the different biological 
pathways is widespread as reflected in registry data and data from recently completed 
clinical trials such as SERAPHIN (Pulido, 2013) and PATENT (Ghofrani, 2013).
Ambrisentan (Volibris) is indicated for treatment of pulmonary arterial hypertension 
(PAH) in adult patients of WHO Functional Class (FC) II to III, including use in 
combination treatment (Volibris, EMA SmPC, Section 5.1). Efficacy has been shown in 
idiopathic PAH (IPAH) and in PAH associated with connective tissue disease.(Volibris, 
EMA SmPC). Ambrisentan is an oral, once daily, propanoic acid-based, ETA-selective 
Endothelin receptor antagonist (ERA) which targets the phospholipase-C-dependent 
endothelin pathway and which is known to play an essential role in mammalian 
cardiovascular physiology.
Tadalafil (Adcirca) is indicated in adults for the treatment of PAH classified as WHO 
functional class II and III, to improve exercise capacity (Adcirca, EMA, SmPC). 
Tadalafil is an oral, once daily, phosphodiesterase type 5(PDE-5) inhibitor which targets
the NO pathway. Through inhibition of PDE-5, tadalafil increases cytoplasmic cGMP 
concentrations in the smooth muscle cells and enhances NO-mediated vasodilatation of 
the vasculature.
An ambrisentan/tadalafil combination therapy is a rational treatment strategy for patients 
with PAH. Both components are orally administered once a day, have different 
mechanisms of action targeting different intracellular pathways, have no clinically 
relevant pharmacokinetic (PK) interactions and are well tolerated when co administered. 
Nonclinical pharmacology data (Liang, 2012) demonstrates a synergistic effect of 
ambrisentan and tadalafil on vasodilatation, whilst a combination of tadalafil and other 
non selective ERAs (bosentan and macitentan) are additive.
A Phase 1 study (GS-US-300-0112) in 26 healthy subjects was performed to detect any 
significant PK interactions between tadalafil and ambrisentan when co-administered 
(Spence, 2009). From this study, it was concluded that there is no clinically significant 
PK interaction between ambrisentan (10 mg) and tadalafil (40 mg) when combined.
Multiple doses of tadalafil had no clinically relevant effect on the PK of either 
ambrisentan or its metabolite, 4-hydroxymethyl ambrisentan. Similarly, the single-dose 
PK of tadalafil were unaffected by multiple doses of ambrisentan. Hence, no dose 
adjustments for ambrisentan or tadalafil should be necessary when these drugs are co-
administered. . There were no SAE’s in the study. Three subjects withdrew due to 
adverse events (AEs): one for anaemia (mild) in the last dosing session on combination, 
following ambrisentan alone and tadalafil alone; one subject because of myalgia (severe), 
muscle fatigue and dizziness on tadalafil alone and one because of headache (severe) on 
the first day of tadalafil and 3 days after ambrisentan. The anaemia was mild and is a 
 
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listed event for ambrisentan. There were a total of 7 subjects with mild tachycardia. There 
were 5 events on ambrisentan 10 mg, 4 days after tadalafil 40 mg. There were 3 events on 
the first day of tadalafil 40 mg given 3 days after ambrisentan 10 mg. There was one 
event on ambrisentan 10 mg and tadalafil 40 mg after 4 days of ambrisentan 10 mg. 
Taken together these data suggest that in healthy subjects a mild tachycardia may result 
from combination use, which is primarily transient. 
Both the marketed products can be taken with or without food (Volibris, EMA SmPC, 
Adcirca, EMA, SmPC).
The AMBITION clinical study (Galiè N, 2015 and AMB112565 CSR 13Nov2014),
which evaluated the time to first clinical failure event, a composite endpoint, shows a 
robust clinical benefit (50% hazard reduction) for PAH patients initiated on a 
combination of ambrisentan and tadalafil when compared to PAH patients initiated on 
either medication as monotherapy.  The safety profile of the combination arm was 
consistent with the known safety data of the individual study drugs and no safety signals 
specific to combination treatment were identified.
ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension (Galie`,
2015) have very recently been updated. The combination of ambrisentan – tadalafil is 
now recommended for efficacy of initial drug combination therapy for pulmonary arterial 
hypertension (group 1) according to World Health Organization functional class I-III.
The treatment of PAH is complex leading to significant patient burden. Patients require 
multiple medications, regular clinical review and repeated clinical assessment. The 
advances in the field, as described, has improved patient outcomes but at the same time 
added further complexity to the management of the disease. Therefore, GSK is proposing 
to develop a fixed dose combination formulation of ambrisentan and tadalafil for the 
treatment of PAH. This will reduce pill burden for patients, which may improve 
treatment compliance and offer a simplified treatment option for both patients and 
physicians. Further, there would be a reduced environmental impact from using a FDC, 
as opposed to separate monotherapies; these would include reduced packaging, storage 
and shipment requirements.This is in accordance with the EMEA guidelines on clinical 
development of fixed dose combination medicinal products (EMEA, 
CHMP/EWP/240/95).
 
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3.
OBJECTIVE(S) AND ENDPOINT(S)
Objectives
Endpoints
Primary
To characterise the relative bioavailability of 
ambrisentan and tadalafil following 
administration of candidate tablet formulations of 
the FDC (ambrisentan 10 mg + tadalafil 40 mg) 
relative to reference monotherapies tested 
(ambrisentan 10 mg & tadalafil 40 mg) taken 
concurrently in healthy human subjects under 
fasting conditions.
Plasma PK parameters: Cmax, AUC(0-), 
and AUC(0-t) of ambrisentan and tadalafil in 
FDC and reference treatments
Secondary
To characterise the secondary PK parameters of 
the FDC and reference treatments in healthy 
human subjects under fasting conditions.
Plasma PK parameters including; tmax, t1/2
of ambrisentan and tadalafil in FDC and 
reference treatments
To characterise the PK parameters of potential 
candidate FDC formulations in healthy human 
subjects under fed and fasted conditions
Plasma PK parameters: Cmax, AUC(0-), 
AUC(0-t), tmax and t1/2 of ambrisentan and 
tadalafil in FDC
To monitor the safety and tolerability of the FDC 
candidate tablets compared to the reference in 
healthy human subjects under fasting conditions.
Safety and Tolerability of all treatments as 
assed by vital signs, ECG, clinical 
laboratory safety data and review of 
adverse events
4.
STUDY DESIGN
4.1.
Overall Design
This is a single centre, Phase 1, single dose, randomised, open label crossover study with 
3 study parts; each study part of the study will be, up to, a 5 way cross over, in healthy 
subjects. See Figure 1 for study schematic.
 
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Figure 1: Study Schematic
All subjects will attend the unit for Screening within 31 days of their first dose. The 
Clinical Unit has an approved panel screening protocol which may be utilised to identify 
eligible study candidates based on the defined study inclusion/ exclusion criteria. Further 
information on requirements for using the approved panel screen protocol is included in 
Section 7.2.
Eligible subjects will be randomised to order of treatment in that study parts in which 
they are included and the order in which these treatments are administered will be in 
accordance with the randomisation schedule.
There will be a minimum washout of 7 days between each dose in a study part.  A 
Follow-Up Visit should be completed within 7-14 days of completion of a subject’s last 
dose. 
The utility of each of the 3 study parts is as follows:
Part 1
This study part, will have up to 5 dose sessions and will be used  to characterise the 
relative bioavailability, safety and tolerability of  up to, four formulations of the fixed 
dose combination (ambrisentan 10 mg +tadalafil 40 mg) and the reference of the 2 
 
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monotherapy components taken concurrently (ambrisentan 10 mg & tadalafil 40 mg). All 
doses will be taken in the fasted state.
If successful formulations are identified in this study part, then they may be reformulated 
and tested in Part 2. If no successful formulations are identified in this study part then 
Part 2 may be utilised to look at up to 4 new FDC formulations.
Part 2
This study part is flexible and will have up to 5 dose sessions. It will be used  to 
characterise, but not limited to, the bioavailability, safety and tolerability of  up to, a 
further, four formulations of the fixed dose combination (ambrisentan 10 mg + tadalafil
40 mg) and the reference of the 2 monotherapy components taken concurrently 
(ambrisentan 10 mg & tadalafil 40 mg). All doses are taken in the fasted state.
However, If only two formulations, or less, are evaluated in Part 2 then the FDC 
formulations may be tested both fed (FDA high fat breakfast) and fasted to assess food 
effect and Part 3 will not be required.
If successful formulations are identified in this study part, then up to 2 of these may be 
tested, for food effect, in Part 3 if not already assessed in this part.
Part 3
Part 3 of the study is optional and utility is dependent on the results of the previous study 
parts. This study part will only be required if formulation to be taken through to a pivotal 
BE study have been identified and the fed and fasted pharmacokinetics of any FDC 
formulations identified for progression have not been tested in Part 2.
This study part will have up to 4 dose sessions and be utilized to access the 
pharmacokinetics, safety and tolerability of up to 2 fixed dose combinations, in both the 
fed and fasted state and which have been identified for progression to the pivotal BE 
study. The fed arms of this part will have the standard FDA high fat breakfast.
Subjects who had taken part in the study part preceding Part 3, and which provided the 
successful formulations for a BE study could also be invited to take part in this study 
part. However, a subject’s inclusion in more than one study part would be dependent on 
the subject not exceeding the maximum blood draw volume (500ml) for the study.
4.2.
Treatment and Duration
The treatments for each study part of the study are listed in Table 1 All treatments are 
single dose. Subjects will be randomised to order of treatments in the parts of the study 
they are included in.
The study has 3 parts and ongoing analyse of pharmacokinetic data will be used to enable 
the formulations produced and tested in subsequent parts.
 
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Part 1
Part 1 of the study will be utilised to look at up to 4 pilot FDC formulations and these are 
described in Section 6.1. Pharmacokinetic data from Part 1 of the study will be analysed 
following completion of the third, fourth and fifth treatment session by subjects. This 
ongoing review of the pharmacokinetic data will enable rapid ongoing assessment of each 
FDC formulation and will be used to enable the formulation development work to 
produce the FDC formulations to be tested in Part 2 of the study.
Successful formulations will be primarily chosen on the test/reference ratio for both AUC 
and Cmax for both components ideally the 90% CI for the ratio would fall in the range of 
0.80 and 1.25, with successful formulations having test/reference ratios close to unity and 
with low intra subject variability indicated by a narrow CI. Any formulations identified 
by these criteria would be reformulated, with the final intended API, for testing in Part 2. 
If no successful formulations are identified from these criteria then the pharmacokinetic 
data would be used to enable further work to produce new FDC formulations for Part 2
Following completion of Part 1 there will be a pause prior to Part 2, so that up to a further 
4 FDC formulations could be produced and data included and approved in any required 
update to submissions to the oversight authorities.
Part 2
Part 2 of the study will be providing data for up to 4 FDC formulations. These could be 
either successful formulation identified in Part 1 reformulated with the final API and/or 
new formulations using the final API. Pharmacokinetic data from Part 2 of the study will 
be analysed following completion of each treatment arm by subjects. This ongoing 
review of the pharmacokinetic data will enable rapid ongoing assessment of each FDC 
formulation and will be used to define any successful FDC formulation to be taken into 
Part 3 of this study and a pivotal BE study. If only two formulations are evaluated in Part 
2 then the food effect may be added to Part 2 and Part 3 will not be required. Success will 
be defined with the same criteria as those in Part 1.
Part 3
Part 3 of the study will provide data for up to 2 successful formulations to be progressed 
to a pivotal BE study, in both the fed and fasted state.
 
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Table 1
Treatments proposed per study part
Part 1 (fasted)
Part 2 (fasted)1
Part 32
ambrisentan and tadalafil
FDC1 (10mg/40mg)
ambrisentan and tadalafil
FDC5 (10mg/40mg)
ambrisentan and tadalafil
FDCX1 (10mg/40mg), fed
ambrisentan and tadalafil
FDC2 (10mg/40mg)
ambrisentan and tadalafil
FDC6 (10mg/40mg)
ambrisentan and tadalafil
FDCX1 (10mg/40mg) fasted
ambrisentan and tadalafil
FDC3 (10mg/40mg)
ambrisentan and tadalafil
FDC7 (10mg/40mg)
ambrisentan and tadalafil
FDCX2 (10mg/40mg), fed
ambrisentan and tadalafil
FDC4 (10mg/40mg)
ambrisentan and tadalafil
FDC8 (10mg/40mg)
ambrisentan and tadalafil
FDCX2 (10mg/40mg), fasted
ambrisentan and tadalafil
monotherapies taken 
concurrently (10mg/40mg)
ambrisentan and tadalafil
monotherapies taken 
concurrently (10mg/40mg)
1.
If 2 or less treatments are tested in Part 2, then these could be tested in both fed and fasted state
2.
If food effect is tested in Part 2 then part 3 may not be required
Due to the flexibility of the protocol there is a range for a subject’s possible duration on 
study. Subjects may be included in Part 1, Part 2 or Part 3 of the study. However, if Part 2 
of the study identifies any successful formulations to take into Part 3 then subjects from 
Part 2 may be invited to join Part 3.
The minimum and maximum duration for a subject on this study is described in Table 2. 
This table assumes all 5 dose sessions are used for Part 1 and Part 2 and either 2 or 4 dose 
sessions are used in Part 3. However, this is an approximation as fewer sessions may be 
required in Part 2 and/or the washout between sessions of 7 days in each part may be 
longer, due to logistical reasons.
 
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Table 2
Study Duration
Study Options
Screen
Study 
Part 
1or 2
Pause
Study 
part 3
Follow-
Up
Visit
Total 
Duration 
on Study
(1-31 
days)
(31 
days)
(7-28 
days)
(10-24 
days)
(7-14 
days)
Subjects 
included 
in only 
Part 1 
or 2
Min
1
31
7
39
Max
31
31
14
76
Subjects 
included 
in Part 
2&3
Min
1
31
7
10
7
56
Max
31
31
28
24
14
128
Subjects 
included 
in only 
Part 3
Min
1
10
7
18
Max
31
24
14
69
A completed subject is one who has completed all study parts they have been randomised 
to and the Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
4.3.
Type and Number of Subjects
A maximum of 20 healthy adult subjects will be randomized, to each study part, such that 
approximately 16 evaluable subjects complete each part of the study.
The study is not powered. However, the number of subjects will be sufficient for the 
objectives of this study and this is detailed further in Section 9.2 Sample Size 
Considerations.
If subjects prematurely discontinue the study, additional replacement subjects may be 
randomised and assigned to the same treatment sequence at the discretion of the Sponsor 
in consultation with the Investigator.
 
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4.4.
Design Justification
The rationale for why ambrisentan and tadalafil should be co-formulated in a new FDC, 
for the treatment of PAH, have been explained in Section 2.2. This study provides the 
first data to enable this and has been designed to identify suitable candidate FDC 
formulations to take into a pivotal BE study and to also provide pharmacokinetic data to 
enable the pivotal BE study to be powered sufficiently. 
The single dose, cross over design, used in each study part, is a standard design and 
sufficient to enable the objectives of the study.
The primary endpoints of the study are pharmacokinetic and as such placebo is not 
warranted, or is there any need to blind study treatment. The Inclusion of the two 
monotherapies taken concurrently provides the reference for the primary pharmacokinetic 
objective and will also provide a comparison for the secondary safety endpoints, when 
compared to any FDC formulations tested.
The study is in three parts to enable PK data from Part 1 to enable formulation 
development for Part 2. Part 3 providing an opportunity to assess food effect in any FDC 
formulations suitable for development and inclusion in the pivotal bioequivalence study. 
The interdependency of the 3 study parts is described in detail in Section 4.2.
4.5.
Dose Justification
The oral dose for the new FDC formulations and the reference treatment proposed for the 
study of, 40mg for tadalafil and 10mg for ambrisentan are the approved maximum doses 
for the drugs and the strength at which these drugs are marketed, for the treatment of 
PAH. These doses will be tested in this protocol and will provide sufficient exposure for 
the pharmacokinetic study endpoints for both components and have previously shown to
be tolerated in healthy subjects.
All treatments are single dose, which is a sufficient duration for assessment of relative 
bioavailability and effect of food on the pharmacokinetics of selected FDC formulations. 
The terminal half life of each component indicates a minimum washout of 7 days,
between doses, is also sufficient for clearance of previous dose.
4.6.
Benefit: Risk Assessment
There is no direct benefit for healthy subjects participating in this study.  The risks to the 
healthy subjects based upon previous experience indicate no expectation of SAE and mild 
to moderate AE.  The benefit/risk remains satisfactory for this healthy subject study.
Summaries of findings from clinical studies conducted with both the Investigational 
Product taken as monotherapies and in combination can be found in:

Ambrisentan IB (Volibris IB, 2015)

Volibris (Ambrisentan) EMA SmPC (Volibris EMA SmPC) 

Adcirca (Tadalafil) EMA SmPC (Adcirca, EMA SmPC)
 
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As described in Section 2.2, mild tachycardia was the most common (7/26, 27%) adverse 
event seen in healthy subjects following combination dosing with 10mg ambrisentan and 
40mg tadalafil (Spence, 2009). There were three subjects (3/26, 11.5%) withdrawn due to
AEs (e.g., anaemia [mild]; myalgia [severe], muscle fatigue, and dizziness; and headache 
[severe]).    No SAEs or additional risks were observed with combination use in healthy 
subjects.
The following section outlines the risk assessment and mitigation strategy for this 
protocol:  
4.6.1.
Risk Assessment
Risks of clinical significance, identified in subjects with PAH and the mitigation strategy
identified for this study in healthy subjects are captured in Table 3. 
 
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Table 3
Risks of Clinical Significance and Mitigation Strategy
Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy
Investigational Product: GSK1325760 (ambrisentan-Volibris)

Teratogenicity

Anaemia (decreased 
haemoglobin, decreased 
haematocrit)

Hypersensitivity reactions 
(e.g. angioedema, rash, 
pruritus)

Headache (including sinus 
headache, migraine)

Dizziness

Cardiac failure

Palpitation

Hypotension

Flushing

Epistaxis

Dyspnoea

Upper respiratory 
congestion, sinusitis, 
nasopharyngitis, rhinitis

Abdominal pain

Constipation

Nausea, vomiting, 
diarrhoea

Hepatic transaminases 
increased

Peripheral oedema, fluid 
retention

Chest pain/discomfort

Asthenia and fatigue
Ambrisentan IB (Volibris IB, 
2015)
Volibris (Ambrisentan) EMA 
SmPC (Volibris, EMA SmPC): 
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision. 

Exclusion of women of 
childbearing potential 
(teratogenicity)

Laboratory assessments 
per protocol

Physical assessment per 
protocol

Routine vital signs per 
protocol

Subjects remain in the 
clinical unit, under medical 
supervision, for all doses 
and until completion of 
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GF196960 (tadalafil - Adcirca)

Hypersensitivity reactions

Headache

Syncope,

Migraine

Blurred vision

Palpitations

Nasopharyngitis (including 
nasal congestion, sinus 
congestion and rhinitis)

Epistaxis
Adcirca (Tadalafil) EMA SmPC 
(Adcirca, EMA SmPC))
Exclusion criteria, ongoing 
safety assessments, study 
design and medical 
supervision.

Routine vital signs per 
protocol

Physical assessment per 
protocol

Subjects remain in the 
clinical unit for all doses, 
under medical supervision 
and until completion of 
 
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Potential Risk of Clinical 
Significance
Summary of Data/Rationale for 
Risk
Mitigation Strategy

Nausea,

Dyspepsia (including 
abdominal pain/discomfort)

Vomiting,

Gastroesophageal reflux

Rash

Myalgia,

Back pain

Pain in extremity (including 
limb discomfort)

Increased uterine bleeding

Facial oedema,

Chest pain
safety assessments at 
48hrs post dose.

Single doses used in the 
study
Investigational Product: GSK3380154 (ambrisentan-tadalafil-FDC)
The risks for the combination 
are the same as the 2 
monotherapies; no extra risks 
have been identified for the 
combination.
As for GSK1325760 and 
GF196960 above
4.6.2.
Benefit Assessment
There is no direct benefit for healthy subjects taking part in this study.  
However, the intended benefit of this study is to inform the correct formulation for a 
fixed dose combination of ambrisentan plus tadalafil to use as first line therapy in PAH 
patients.
4.6.3.
Overall Benefit: Risk Conclusion
Though there is no direct benefit for healthy subjects, based on observations from 
previous healthy subject studies, the adverse event burden is mild, consistent with 
observations in patients and all risks have been mitigated as described in Table 3. So, the 
risk: benefit is appropriate for this study and to enable development of this investigational 
product. The overall benefit: risk therefore remains positive.
5.
SELECTION OF STUDY POPULATION AND 
WITHDRAWAL CRITERIA
Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in:

Volibris (Ambrisentan) EMA SmPC (Volibris, EMA SmPC)

Adcirca (Tadalafil) EMA SmPC (Adcirca, EMA SmPC)
 
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Deviations from inclusion and exclusion criteria are not allowed because they can 
potentially jeopardize the scientific integrity of the study, regulatory acceptability or 
subject safety. Therefore, adherence to the criteria as specified in the protocol is essential.
5.1.
Inclusion Criteria
A subject will be eligible for inclusion in this study only if all of the following criteria 
apply:
AGE
1. Between 18 and 60 years of age inclusive, at the time of signing the informed 
consent.
TYPE OF SUBJECT AND DIAGNOSIS INCLUDING DISEASE SEVERITY
2. Healthy as determined by a responsible and experienced physician, based on a 
medical evaluation including medical history, physical examination, laboratory tests, 
vital signs and cardiac monitoring (ECG and 24 hour Holter).  A subject with a 
clinical abnormality or laboratory parameter(s) which is/are not specifically listed in 
the inclusion or exclusion criteria, outside the reference range for the population 
being studied may be included only if the Investigator, in consultation with the GSK 
Medical Monitor if required, judges and documents that the finding is unlikely to 
introduce additional risk factors and will not interfere with the study procedures. 
WEIGHT
3. Body weight ≥ 50 kg (110 lbs) for men and ≥ 45 kg (99lbs) for women and body 
mass index (BMI) within the range 18 – 30 kg/m2 (inclusive)
SEX
4. Male or Female
Females must be the following:
Non-reproductive potential defined as:

Pre-menopausal females with one of the following:
Documented tubal ligation
Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion 
Hysterectomy
Documented Bilateral Oophorectomy

Documented Postmenopausal defined as 12 months of spontaneous amenorrhea
 
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INFORMED CONSENT
5. Capable of giving signed informed consent as described in Section 10 which includes 
compliance with the requirements and restrictions listed in the consent form and in 
this protocol.
5.2.
Exclusion Criteria
A subject will not be eligible for inclusion in this study if any of the following criteria 
apply:
CONCURRENT CONDITIONS/MEDICAL HISTORY (INCLUDES LIVER 
FUNCTION AND QTC INTERVAL)
1.
A blood pressure <100/55 mm Hg.
2.
Haemoglobin below normal range:

Hb < 133 g/L for males 

Hb< 114 g/L for females
3.
ALT and bilirubin >1.5xULN (isolated bilirubin >1.5xULN is acceptable if bilirubin 
is fractionated and direct bilirubin <35%). 
4.
Current or chronic history of liver disease, or known hepatic or biliary abnormalities 
(with the exception of Gilbert's syndrome or asymptomatic gallstones)
5.
QTc> 450 msec
NOTES:

The QTc is the QT interval corrected for heart rate according to Bazett’s 
formula (QTcB), Fridericia’s formula (QTcF), and/or another method, machine-
read or manually over-read.

The specific formula that will be used to determine eligibility and 
discontinuation for an individual subject should be determined prior to initiation 
of the study. In other words, several different formulae cannot be used to 
calculate the QTc for an individual subject and then the lowest QTc value used 
to include or discontinue the subject from the trial.

For purposes of data analysis, QTcB, QTcF, another QT correction formula, or a 
composite of available values of QTc will be used as specified in the Reporting 
and Analysis Plan (RAP).
 
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CONCOMITANT MEDICATIONS
6.
Use of prescription or non-prescription drugs, including vitamins, herbal and dietary 
supplements (including St John’s Wort) within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose 
of study medication, unless in the opinion of the Investigator and GSK Medical 
Monitor the medication will not interfere with the study procedures or compromise 
subject safety. 
RELEVANT HABITS
7.
History of regular alcohol consumption within 6 months of the study defined as: 

An average weekly intake of >21 units for males or >14 units for females.  One 
unit is equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 
ml) of wine or 1 (25 ml) measure of spirits.
8.
Smoking more than 5 cigarettes per week and subjects must be able to abstain from 
smoking for a 24 hour period prior to dose and any time whilst in the clinical unit.
CONTRAINDICATIONS
9.
History of sensitivity to any of the study medications, or components thereof or a 
history of drug or other allergy that, in the opinion of the Investigator or Medical 
Monitor, contraindicates their participation.
DIAGNOSTIC ASSESSMENTS AND OTHER CRITERIA
10. Presence of hepatitis B surface antigen (HBsAg), positive hepatitis C antibody test 
result at Screening or within 3 months prior to first dose of study treatment. . 
11. A positive test for HIV antibody.
12. A positive pre-study drug/alcohol screen.  
13. Where participation in the study would result in donation of blood or blood products 
in excess of 500 mL within previous 3 months
14. The subject has participated in a clinical trial and has received an investigational 
product within the following time period prior to the first dosing day in the current 
study: 3 months, 5 half-lives or twice the duration of the biological effect of the 
investigational product (whichever is longer).
15. Exposure to more than four new chemical entities within 12 months prior to the first 
dosing day.
 
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5.3.
Screening/Baseline/Run-in Failures
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT) 
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.4).
5.4.
Withdrawal/Stopping Criteria
The following actions must be taken in relation to a subject who fails to attend the clinic 
for a required study visit:

The site must attempt to contact the subject and re-schedule the missed visit as 
soon as possible. 

The site must counsel the subject on the importance of maintaining the assigned 
visit schedule and ascertain whether or not the subject wishes to and/or should 
continue in the study. 

In cases where the subject is deemed ‘lost to follow up’, the investigator or 
designee must make every effort to regain contact with the subject (where 
possible, 3 telephone calls and if necessary a certified letter to the subject’s last 
known mailing address or local equivalent methods). These contact attempts 
should be documented in the subject’s medical record. 

Should the subject continue to be unreachable, only then will he/she be considered 
to have withdrawn from the study with a primary reason of “Lost to Follow-Up”. 
A subject may withdraw from study treatment at any time at his/her own request, or may 
be withdrawn at any time at the discretion of the investigator for safety, behavioural or 
administrative reasons. If a subject withdraws from the study, he/she may request 
destruction of any samples taken, and the investigator must document this in the site 
study records. 
5.4.1.
Liver Chemistry Stopping Criteria
Liver chemistry stopping and increased monitoring criteria have been designed to 
assure subject safety and evaluate liver event etiology (in alignment with the FDA 
premarketing clinical liver safety guidance). 
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf
 
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Study treatment will be discontinued for a subject if liver chemistry stopping criteria are 
met:
Phase I Liver Chemistry Stopping Criteria – Liver Stopping Event Algorithm
Continue Study Treatment 
Discontinue Study Treatment 
*INR value not applicable to subjects on anticoagulants 
ALT ≥3xULN 
No
Yes
Must refer to Liver Safety Required Actions and Follow up Assessments  section in the Appendix
Report as an SAE if possible Hy’s Law case: ALT≥3xULN and  Bilirubin≥2xULN (>35% direct)  or 
INR>1.5, if measured*
Liver Safety Required Actions and Follow-Up Assessments Section can be found in 
Appendix 2: Liver Safety Required Actions and Follow-Up Assessments
5.4.1.1.
Study Treatment Restart or Rechallenge
Study treatment restart or re-challenge after liver chemistry stopping criteria are met by 
any subject participating in this study is not allowed.
5.4.2.
QTc Stopping Criteria

The same QT correction formula must be used for each individual subject to 
determine eligibility for and discontinuation from the study.  This formula may 
not be changed or substituted once the subject has been enrolled.

For example, if a subject is eligible for the protocol based on QTcF, then QTcF
must be used for discontinuation of this individual subject as well.

Once the QT correction formula has been chosen for a subject’s eligibility, the 
same formula must continue to be used for that subject for all QTc data being 
collected for data analysis.  Safety ECGs and other non-protocol specified 
ECGs are an exception.  

The QTc should be based on averaged QTc values of triplicate electrocardiograms 
obtained over a brief (e.g., 5-10 minute) recording period. 
A subject that meets either bulleted criterion below will be withdrawn from the study.  

QTc> 500 msec,
 
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
Change from baseline: QTc >60 msec 
5.4.3.
Hypotension
A subject that meets bulleted criterion below will be withdrawn from the study.

For hypotension: if systolic <90 or diastolic <50 confirmed by triplicate reading 
taken upto 5 minutes apart.
5.4.4.
Other Dose Adjustment/Stopping Safety Criteria
For an individual study participant, stopping criteria include, but are not limited to:
Adverse events, or significant changes in any of the safety assessments, that put the 
safety of the individual at risk (e.g., ECG, vital signs, laboratory tests, etc), as judged by 
the Principal Investigator in consultation with the Medical Monitor if necessary.
5.5.
Subject and Study Completion
A completed subject is one who has completed all phases of the study including the 
Follow-Up visit.
The end of the study is defined as the last subject’s last visit.
6.
STUDY TREATMENT
6.1.
Investigational Product and Other Study Treatment
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments. Study treatments for Part 1 of the study are shown in Table 4, this will be 
amended once treatments for Part 2 and Part 3 are confirmed.
 
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Table 4
Study Treatments for Part 1
Study Treatment
Product name:
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC)
GSK3380154 
(ambrisentan-
tadalafil-FDC) 
GSK1325760 
(ambrisentan-Volibris)
GF196960 
(tadalafil - Adcirca)
Treatment
FDC1
FDC2
FDC3
FDC4
Reference
Formulation 
description
GSK3380154, TAB-
A, Tablet Weight 
840mg/2mg SLS*
GSK3380154, TAB-A, 
Tablet Weight 
840mg/4mg SLS*
*
GSK3380154, TAB-
A, Tablet Weight 
560mg/2mg SLS*
GSK3380154, 
TAB-A, Tablet 
Weight 
560mg/4mg SLS
Each tablet contains 10 
mg of ambrisentan, 
approximately 95 mg of 
lactose (as 
monohydrate), 
approximately 0.25 mg 
of lecithin (soya) (E322) 
and approximately 0.11 
mg of Allura red AC 
Aluminium Lake 
Each film-coated 
tablet contains 20 
mg tadalafil, 233 mg 
lactose (as 
monohydrate).
Dosage form:
Film-coated tablet
Film-coated tablet
Film-coated tablet
Film-coated 
tablet
Film-coated tablet
Film-coated tablet
Unit dose 
strength(s)/
Dosage 
level(s):
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 
40 mg tadalafil and 
10 mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet 
contains 40 mg 
tadalafil and 10 
mg of 
ambrisentan.
Each tablet contains 10 
mg of ambrisentan.
Each tablet contains 
20 mg tadalafil.
Route of 
Administration
Oral
Oral
Oral
Oral
Oral
Oral
Dosing 
instructions
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
Take as directed
 
 
 
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6.2.
Medical Devices
No GSK manufactured devices (or devices manufactured for GSK by a third party) are 
provided for use in this study.
6.3.
Treatment Assignment
Subjects will be assigned to treatment sequence, for the study part/s that they are included 
in and in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal software.
The treatments are denoted as F1 to F4 for Part 1, F5 to F8 for Part 2 and R for reference 
treatment for both parts; the selected formulations for Part 3 are denoted as X and Y.  The
treatment key for Parts 1, 2 and 3 are described in Table 5
Table 5
Treatment Key for Part 1, Part 2 and Part 3
Treatment
Description
Part 1
F1
ambrisentan and tadalafil FDC1 (10mg/40mg)
F2
ambrisentan and tadalafil FDC2 (10mg/40mg)
F3
ambrisentan and tadalafil FDC3 (10mg/40mg)
F4
ambrisentan and tadalafil FDC4 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 2
F5
ambrisentan and tadalafil FDC5 (10mg/40mg)
F6
ambrisentan and tadalafil FDC6 (10mg/40mg)
F7
ambrisentan and tadalafil FDC7 (10mg/40mg)
F8
ambrisentan and tadalafil FDC8 (10mg/40mg)
R
ambrisentan and tadalafil monotherapies taken concurrently 
(10mg/40mg)
Part 3
X1
ambrisentan and tadalafil FDCX1 (10mg/40mg), fed
X2
ambrisentan and tadalafil FDCX1 (10mg/40mg) fasted
Y1
ambrisentan and tadalafil FDCX2 (10mg/40mg), fed
Y2
ambrisentan and tadalafil FDCX2 (10mg/40mg), fasted
The treatment sequence assignments for each part of the study, based on the Latin 
Squares for Williams Designs are shown in Table 6. Not all possible sequences are 
included here. Such as, if only one formulation taken into Part 2 or 3, or Part 2 is used for 
 
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only one or two FDC formulations, fed and fasted. Additional treatment sequences will 
be created based on the Latin Squares for Williams Designs, as required.
Table 6
Treatment Sequence assignments for Part 1, Part 2 and Part 3
Study 
Parts
Number of Treatments
Sequence assignments
Allocation ratio
Part 1
5
F1 F2 R F3 F4
F2 F3 F1 F4 R
F3 F4 F2 R F1,
F4 R F3 F1 F2
R F1 F4 F2 F3,
F4 F3 R F2 F1
R F4 F1 F3 F2,
F1 R F2 F4 F3,
F2 F1 F3 R F4,
F3 F2 F4 F1 R
1:1:1:1:1:1:1:1:1:1
Part 2
5
F5 F6 R F7 F8
F6 F7 F5 F8 R
F7 F8 F6 R F5
F8 R F7 F5 F6
R F5 F8 F6 F7
F8 F7 R F6 F5
R F8 F5 F7 F6
F5 R F6 F8 F7
F6 F5 F7 R F8
F7 F6 F8 F5 R
1:1:1:1:1:1:1:1:1:1
4
F5 F6 R F7
F6 F7 F5 R
F7 R F6 F5
R F5 F7 F6
1:1:1:1
3
F5 F6 R
R F6 F5
F6 R F5
F1 R F6
R F5 F6
F6 F5 R
3:3:3:3:3:3 + 2 
random
2
F5 R 
R F5
1:1
Part 3
4
X1 X2 Y2 Y1
X2 Y1 X1 Y2
Y1 Y2  X2 X1
Y2  X1 Y1 X2 
1:1:1:1
 
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6.4.
Blinding
This will be an open-label study. 
6.5.
Packaging and Labeling
The contents of the label will be in accordance with all applicable regulatory 
requirements.
6.6.
Preparation/Handling/Storage/Accountability
This will be detailed in a Study Specific Technical Agreement/Memo (TTS) which will 
be accompanied by a Quality Agreement.

Only subjects enrolled in the study may receive study treatment and only 
authorized site staff may supply or administer study treatment.  All study 
treatments must be stored in a secure environmentally controlled and monitored 
(manual or automated) area in accordance with the labelled storage conditions 
with access limited to the investigator and authorized site staff. 

The investigator, institution, or the head of the medical institution (where 
applicable) is responsible for study treatment accountability, reconciliation, and 
record maintenance (i.e. receipt, reconciliation and final disposition records).  

Further guidance and information for final disposition of unused study treatment 
are provided in the SRM.

Under normal conditions of handling and administration, study treatment is not 
expected to pose significant safety risks to site staff.  

A Material Safety Data Sheet (MSDS)/equivalent document describing 
occupational hazards and recommended handling precautions either will be 
provided to the investigator, where this is required by local laws, or is available 
upon request from GSK.
6.7.
Compliance with Study Treatment Administration
When subjects are dosed at the site, they will receive study treatment directly from the 
investigator or designee, under medical supervision.  The date and time of each dose 
administered in the clinic will be recorded in the source documents.  The dose of study 
treatment and study subject identification will be confirmed at the time of dosing by a 
member of the study site staff other than the person administering the study treatment.  
Study site personnel will examine each subject’s mouth to ensure that the study treatment 
was ingested.
6.8.
Treatment of Study Treatment Overdose
For this study, any dose of GSK3380154 (ambrisentan/tadalafil-FDC), GSK1325760
(ambrisentan) and GF196960 (tadalafil) greater than the protocol defined dose and within 
a 24 hour time period will be considered an overdose.
 
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GSK does not recommend specific treatment for an overdose.
Advice for the investigator is included in the product label for GSK1325760 
(ambrisentan) and GF196960 (tadalafil).
In the event of an overdose the investigator should:
1.
Contact the Medical Monitor immediately
2.
Closely monitor the subject for adverse events (AEs)/serious adverse events (SAEs) 
and laboratory abnormalities until compound number/name can no longer be 
detected systemically (at least 3 days for compound number/name)
3.
Obtain a plasma sample for pharmacokinetic (PK) analysis if requested by the 
Medical Monitor (determined on a case-by-case basis)
4.
Document the quantity of the excess dose as well as the duration of the overdosing in 
the CRF.
Decisions regarding dose interruptions or modifications will be made by the investigator 
in consultation with the Medical Monitor based on the clinical evaluation of the subject.
6.9.
Treatment after the End of the Study
Subjects will not receive any additional treatment from GSK after completion of the 
study because only healthy subjects are eligible for study participation.
6.10.
Lifestyle and/or Dietary Restrictions
6.10.1.
Meals and Dietary Restrictions

Refrain from consumption of red wine, Seville oranges, grapefruit or grapefruit 
juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or fruit juices from 
7 days prior to the first dose of study medication until after the final dose.

Dependent on the utility of the dose session subjects will be asked to fast or 
consume the FDA full fat breakfast prior to dosing:

Fasting subjects will be required to fast from midnight before each full in-house 
dosing day (i.e. Day 1) with the exception of water, which will be allowed freely
except for 1 hour either side of dosing. Subjects will be required to fast up to 4h 
hour post dose on Day 1 of each dose

For treatments given under fed conditions, a standard high-fat breakfast will be 
provided before dosing. See Section 12.4 for details of this meal. This meal 
should be eaten in its entirety within 30 minutes.  The amount consumed will be 
recorded within the source documents and the CRF by the site staff.  Subjects 
will be dosed within 5 minutes of completing the breakfast.  

Subjects should take each dose of investigational product with 240 ml (8 fl oz) of 
water.
 
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6.10.2.
Caffeine, Alcohol, and Tobacco

During each dosing session, subjects will abstain from ingesting caffeine- or 
xanthine-containing products (e.g., coffee, tea, cola drinks and chocolate) for 24
hours prior to the start of dosing until collection of the final pharmacokinetic sample 
during each session. 

During each dosing session, subjects will abstain from alcohol for 24 hours prior to 
the start of dosing until collection of the final pharmacokinetic sample during each 
session. 

Smoking is not allowed for 24 hours prior to dosing and whilst subjects are in the 
clinic.
6.10.3.
Activity
Subjects will abstain from strenuous exercise from Screening until Follow-Up.  Subjects 
may participate in light recreational activities during studies (e.g., watch television, read).
6.11.
Concomitant Medications and Non-Drug Therapies
6.11.1.
Permitted Medications and Non-Drug Therapies
Paracetamol at doses of 2 grams/day is permitted for use any time during the study. 
Other concomitant medication may be considered on a case by case basis by the 
Investigator in consultation with the Medical Monitor if required.
6.11.2.
Prohibited Medications and Non-Drug Therapies
Subjects must abstain from taking prescription or non-prescription drugs (including 
vitamins and dietary or herbal supplements), within 7 days (or 14 days if the drug is a 
potential enzyme inducer) or 5 half-lives (whichever is longer) prior to the first dose of 
study medication until completion of the Follow-Up visit, unless in the opinion of the 
Investigator and sponsor the medication will not interfere with the study.
7.
STUDY ASSESSMENTS AND PROCEDURES
Protocol waivers or exemptions are not allowed with the exception of immediate safety 
concerns. Therefore, adherence to the study design requirements, including those 
specified in the Time and Events Table, are essential and required for study conduct.
This section lists the procedures and parameters of each planned study assessment.  The 
exact timing of each assessment is listed in the Time and Events Table Section 7.1
The following points must be noted: 

If assessments are scheduled for the same nominal time, THEN the assessments 
should occur in the following order:
1. 12-lead ECG
2. Vital signs
 
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3. Blood draws.  
Note: The timing of the assessments should allow the blood draw to occur at the 
exact nominal time.

The timing and number of planned study assessments, safety and 
pharmacokinetic assessments may be altered during the course of the study 
based on newly available data (e.g., to obtain data closer to the time of peak 
plasma concentrations) to ensure appropriate monitoring. 

The change in timing or addition of time points for any planned study 
assessments must be documented in a Note to File which is approved by the 
relevant GSK study team member and then archived in the study sponsor and 
site study files, but this will not constitute a protocol amendment. 

The IRB/IEC will be informed of any safety issues that require alteration of the 
safety monitoring scheme or amendment of the Informed Consent Form. 
No more than 500 mL of blood will be collected over the duration of the study, including 
any extra assessments that may be required.
 
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7.1.
Time and Events Table
Procedure
Screen
Part 1, 2 and 3. Each dose in each Part repeats this schedule.
FU
Notes
Day
≤-31
-1
1
2
3
4
≥7-14
Time (hrs)
Pre-
dose
0
0.5
1
1.5
2
2.5
4
8
12
18
24
36
48
72
Outpatient visit
x
x
x
Admission to unit
x
Informed consent
x
Inclusion and exclusion criteria
x
x
Demography
x
Full physical exam including 
height and weight
x
Brief Physical
x
Medical history (includes 
substance usage)
x
HIV, Hep B and Hep C screen] 
x
Laboratory assessments (include 
liver chemistries) 
x
x
x
x
Only Screening labs need to be taken in 
fasted state.
Serum hCG Pregnancy test
x
x
Female subjects only
Urine hCG Pregnancy test
x
Female subjects only
Breathalyser and Smokerlyzer
x
x
DOA testing
x
x
12-lead ECG
x
x
x
x
x
x
x
x
x
Triplicate at screen and baseline, single 
measure at other times, unless out of 
range then triplicates should be 
performed
Vital signs
x
x
x
x
x
x
x
x
x
x
x
x
24hr Holter
x
Randomisation
x
Randomised prior to first dose only
Study Treatment
x
AE/SAE review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
SAEs from Screen. AEs from first dose
Concomitant medication review
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
x
PK Sample
x
x
x
x
x
x
x
x
x
x
x
x
x
x
Discharge from Unit
x
For logistical reasons subjects may 
remain in-unit for the 72 hr assessments 
if they prefer.
 
 
 
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7.2.
Screening and Critical Baseline Assessments
All subjects must give written consent to participate in this trial.  Consent for screening 
evaluations may be obtained using the ICF for the Hammersmith Medicines Research 
(HMR) healthy subject’s panel, which has been approved by the HRA’s Phase 1 Advert 
Review group.  The study-specific information and consent form will be signed by the
subject either before any screening evaluation or after the investigator confirms the 
eligibility of the subject for the study and before the subject is randomised to receive the 
first administration of IMP.  Before giving consent, subjects must read the information 
sheet about the study.  They must also read the consent form.  They will then discuss the 
study with the investigator or his deputy and be given the opportunity to ask questions.  
The study-specific information sheet and the consent form must be approved by the REC.
7.2.1.
Demographic/Medical History Assessments
Prior to enrolling in the study and having any study procedures completed, subjects must 
sign the informed consent.
During the Screening visit, each subject will undergo the assessments to determine 
eligibility for enrolment as detailed in Section 5. 
The following demographic parameters will be captured: date of birth, gender, race and 
ethnicity.
Medical/medication/alcohol history will be assessed as related to the eligibility criteria 
listed in Section 5.
7.3.
Pharmacokinetics
7.3.1.
Blood Sample Collection
Blood samples for pharmacokinetic (PK) analysis of ambrisentan and tadalafil will be 
collected at the time points indicated in Section 7.1, Time and Events Table.  The actual 
date and time of each blood sample collection will be recorded.  The timing of PK 
samples may be altered and/or PK samples may be obtained at additional time points to
ensure thorough PK monitoring.
For analysis of ambrisentan 2.7 mL of blood will be collected into sodium citrate tubes 
and for analysis of tadalafil 2.0 mL of blood will be collected into K2-EDTA tubes. 
Processing, storage and shipping procedures are provided in the Study Reference Manual 
(SRM).
7.3.2.
Sample Analysis
Plasma analysis will be performed under the control of PTS-DMPK/Scinovo, 
GlaxoSmithKline, the details of which will be included in the Study Reference Manual 
(SRM). Concentrations of ambrisentan and tadalafil will be determined in plasma
 
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samples using the currently approved bioanalytical methodology. Raw data will be 
archived at the bioanalytical site (detailed in the SRM).
Once the plasma has been analyzed for ambrisentan and tadalafil any remaining 
plasma may be analyzed for other compound-related metabolites and the results reported 
under a separate PTS-DMPK/Scinovo, GlaxoSmithKline protocol.
7.4.
Safety
Planned time points for all safety assessments are listed in the Time and Events Table 
(Section 7.1).  Additional time points for safety tests such as vital signs, physical 
examinations and laboratory safety assessments may be added during the course of the 
study based on newly available data to ensure appropriate safety monitoring.
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)
The definitions of an AE or SAE can be found in Appendix 3: Definition of and 
Procedures for Recording, Evaluating, Follow-Up and Reporting of Adverse Events.
The investigator and their designees are responsible for detecting, documenting and 
reporting events that meet the definition of an AE or SAE. 
7.4.1.1.
Time period and Frequency for collecting AE and SAE information

Any SAEs assessed as related to study participation (e.g., protocol-mandated 
procedures, invasive tests, or change in existing therapy) or related to a GSK 
product will be recorded from the time a subject consents to participate in the 
study up to and including any follow up contact. 

AEs will be collected from the start of Study Treatment until the follow up contact
(see Section 7.4.1.3), at the timepoints specified in the Time and Events Table 
(Section 7.1).

Medical occurrences that begin prior to the start of study treatment but after 
obtaining informed consent may be recorded on the Medical History/Current 
Medical Conditions section of the CRF.

All SAEs will be recorded and reported to GSK within 24 hours, as indicated in 
Section 12.3.

Investigators are not obligated to actively seek AEs or SAEs in former study 
subjects. However, if the investigator learns of any SAE, including a death, at any 
time after a subject has been discharged from the study, and he/she considers the 
event reasonably related to the study treatment or study participation, the 
investigator must promptly notify GSK.
NOTE: The method of recording, evaluating and assessing causality of AEs and SAEs 
plus procedures for completing and transmitting SAE reports to GSK are provided in
Section 12.3.
 
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7.4.1.2.
Method of Detecting AEs and SAEs
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended 
and non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrence.  Appropriate questions include:

“How are you feeling?” 

“Have you had any (other) medical problems since your last visit/contact?”

“Have you taken any new medicines, other than those provided in this study,
since your last visit/contact?
7.4.1.3.
Follow-up of AEs and SAEs
After the initial AE/SAE report, the investigator is required to proactively follow each 
subject at subsequent visits/contacts.  All SAEs, and non-serious AEs of special interest 
(as defined in Section 4.6.1) will be followed until resolution, until the condition 
stabilizes, until the event is otherwise explained, or until the subject is lost to follow up 
(as defined in Section 5.4). Further information on follow up procedures is given in Study 
Assessments and Procedures.
7.4.1.4.
Regulatory Reporting Requirements for SAEs
Prompt notification by the investigator to GSK of SAEs related to study treatment (even 
for non- interventional post-marketing studies) is essential so that legal obligations and 
ethical responsibilities towards the safety of subjects and the safety of a product under 
clinical investigation are met. 
GSK has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of a product under clinical investigation.  GSK will 
comply with country specific regulatory requirements relating to safety reporting to the 
regulatory authority, Institutional Review Board (IRB)/Independent Ethics Committee 
(IEC) and investigators.
Investigator safety reports are prepared for suspected unexpected serious adverse 
reactions according to local regulatory requirements and GSK policy and are forwarded 
to investigators as necessary.
An investigator who receives an investigator safety report describing a SAE(s) or other 
specific safety information (e.g., summary or listing of SAEs) from GSK will file it with 
the IB and will notify the IRB/IEC, if appropriate according to local requirements.
7.4.2.
Physical Exams

A complete physical examination will include, at a minimum, assessment of the 
Cardiovascular, Respiratory, Gastrointestinal and Neurological systems. Height 
and weight will also be measured and recorded.

A brief physical examination will include, at a minimum assessments of the skin,
lungs, cardiovascular system, and abdomen (liver and spleen).
 
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
Investigators should pay special attention to clinical signs related to previous 
serious illnesses
7.4.3.
Vital Signs

Vital signs will be measured in semi-supine position after 5 minutes rest and will 
include temperature, systolic and diastolic blood pressure and pulse rate and 
respiratory rate. 

Three readings of blood pressure and pulse rate will be taken at Screening and 
baseline (pre-dose). All subsequent assessments will be single measures, unless 
the subjects blood pressure or pulse rate has changed from baseline by >15% and 
then 2 further readings should be taken and recorded:

For triplicate readings:

All 3 readings will be recorded in the CRF

First reading should be rejected

Second and third readings should be averaged to give the measurement to 
be used and this will also be recorded in the CRF
7.4.4.
Electrocardiogram (ECG)

ECGs will be measured in semi-supine position after 5 minutes rest

Triplicate 12-lead ECGs will be obtained at Screening and baseline (predose). All 
subsequent assessments will be a single measures, unless withdrawal criteria are 
meet and in which case 2 further readings should be taken to confirm if 
withdrawal is required . An ECG machine that automatically calculates the heart 
rate and measures PR, QRS, QT, and QTc.  Refer to Section 5.4.2 for QTc 
withdrawal criteria and additional QTc readings that may be necessary.

24 hour continuous cardiac telemetry (Holter) will be performed at Screening.  
Full disclosures will be reviewed in detail and the review maintained as part of the 
subject’s source documents
7.4.5.
Clinical Safety Laboratory Assessments
All protocol required laboratory assessments, as defined in Table 7, must be conducted in 
accordance with the Laboratory Manual, and Protocol Time and Events Schedule.
Laboratory requisition forms must be completed and samples must be clearly labelled 
with the subject number, protocol number, site/centre number, and visit dat. Reference 
ranges for all safety parameters will be provided to the site by the laboratory responsible 
for the assessments.
If additional non-protocol specified laboratory assessments are performed at the 
institution’s local laboratory and result in a change in subject management or are 
considered clinically significant by the investigator (e.g., SAE or AE or dose 
modification) the results must be recorded in the CRF.
 
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Refer to the SRM for appropriate processing and handling of samples to avoid duplicate 
and/or additional blood draws.
All study-required laboratory assessments will be performed by a local laboratory.
Haematology, clinical chemistry, urinalysis and additional parameters to be tested are 
listed in Table 7.
Table 7
Protocol Required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Haematology
Platelet Count
RBC Indices:
WBC count with Differential:
RBC Count
MCV
Neutrophils
Hemoglobin
MCH
Lymphocytes
Hematocrit
MCHC
Monocytes
Eosinophils
Basophils
Clinical 
Chemistry 1
BUN
Potassium
AST (SGOT)
Total and direct bilirubin
Creatinine
Sodium
ALT (SGPT)
Total Protein
Glucose 
Calcium
Alkaline phosphatise
Albumin
Routine 
Urinalysis3
As a minimum, but not limited to the following tests and dependent on 
standard urinalysis dipstick used:

Specific gravity

pH, glucose, protein, blood and ketones by dipstick

Microscopic examination (if blood or protein is abnormal)
Other 
Screening
Tests

HIV

Hepatitis B (HBsAg)

Hepatitis C (Hep C antibody)

Alcohol and drug screen (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines)

Serum  or urine hCG Pregnancy test2
NOTES :
1.
Details of Liver Chemistry Stopping Criteria and Required Actions and Follow-Up
Assessments after liver stopping or monitoring event are given in Section 5.4.1 and 
Appendix 2
2.
Local urine testing will be standard for the protocol unless serum testing is required 
by local regulation or ethics committee.
3.
Routine Urinalysis results will not be databased, unless a result is out of range and 
clinically significant then it would be captured as an AE  
 
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All laboratory tests with values that are considered clinically significantly abnormal 
during participation in the study or within7 days after the last dose of study treatment 
should be repeated until the values return to normal or baseline.  If such values do not 
return to normal within a period judged reasonable by the investigator, the etiology
should be identified and the sponsor notified.
8.
DATA MANAGEMENT

Data will be double-entered into a clinical database management system 
(ClinPlus Version 3.3).

Management of clinical data will be performed in accordance with applicable 
HMR standards and data cleaning procedures to ensure the integrity of the data, 
e.g., removing errors and inconsistencies in the data.

Adverse events and concomitant medications terms will be coded using 
MedDRA (Medical Dictionary for Regulatory Activities) and an internal 
validated medication dictionary, GSKDrug.  

Original CRFs will be retained by GSK, while HMR will retain a copy. Subject 
initials will not be collected or transmitted to GSK according to GSK policy 
9.
STATISTICAL CONSIDERATIONS AND DATA 
ANALYSES
9.1.
Hypotheses
No formal hypothesis will be tested. An estimation approach will be used to (i) estimate 
the bioavailability of the FDC formulations relative to the monotherapies taken 
concurrently in Part 1 and if used, Study part 2, (ii) estimate the bioavailability of the 
formulation(s) of the FDC formulations, taken in to Part 3, in the fed state relative to the 
fasting state.
For each primary pharmacokinetic endpoint, point estimates and corresponding 90% 
confidence intervals will be constructed for the ratio of the geometric mean of the test 
treatment (FDC formulations) to the geometric mean of the reference treatment (co-
administration of ambrisentan and tadalafil), (test)/ (reference).
9.2.
Sample Size Considerations
No formal sample size calculation has been performed. Sample sizes chosen for each 
study part in this study is considered sufficient for exploratory analysis based on PK 
variability data from previous studies
9.2.1.
Sample Size Assumptions
The sample size assumptions are based on previously reported estimates of within subject 
CV for AUC(0-∞) and Cmax for ambrisentan (GS-US-300-0112) and tadalafil (Forgue, 
2005). Table 8 summarizes the estimates of within subject CV for the primary endpoints 
AUC (0-∞) and Cmax.
 
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Table 8
Estimates of within subject CV for the primary end points AUC (0-∞) 
and Cmax
CVw: within subject CV
ambrisentan
tadalafil
Cmax
22%
16%
AUC (0-∞)
15%
13%
The largest of the within subject CV estimates is about 22%, which translate to a standard 
deviation (SD) of 0.217 on the natural log scale.  Based on this SD, the half width of the 
90% confidence interval will be 12.5% of the point estimate based on a sample size of 20 
statistically evaluable subjects. 
9.2.2.
Sample Size Sensitivity
Assuming the within subject CV differs from 22% and the number of statistically 
evaluable subjects is lower than 20, then Table 9 shows the precision (the half width of 
the 90% confidence interval) for a range of evaluable subjects.
Table 9
Sample Size Sensitivity
Evaluable subjects
CVw
Precision
20
22%
12.5%
20
34%
19.6%
18
22%
13.2%
18
32%
19.6%
16
22%
14.2%
16
30%
19.6%
9.2.3.
Sample Size Re-estimation or Adjustment
No sample size re-estimation will be performed.
9.3.
Data Analysis Considerations
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacokinetics 
Modeling and Simulation Department, CPDM, GlaxoSmithKline. Plasma concentration-
time data for ambrisentan and tadalafil will be analyzed by non-compartmental methods 
with WinNonlin [version 6.3 or above].  Calculations will be based on the actual 
 
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sampling times recorded during the study.  From the plasma concentration-time data, the 
following pharmacokinetic parameters will be determined, as data permit: maximum 
observed plasma concentration (Cmax), time to Cmax (tmax), area under the plasma 
concentration-time curve [AUC(0-t) and AUC(0-∞)], and apparent terminal phase half-
life (t1/2).  
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals R&D.
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline.
Systemic exposure of ambrisentan and tadalafil will be evaluated when administered as 
the FDC compared to administration as the monotherapies concurrently, using loge-
transformed AUC(0-) and Cmax in a mixed effects model.
9.3.1.
Analysis Populations
9.3.1.1.
Safety Population
All subjects enrolled into the study who have received at least one dose of investigational 
product will be included in the Safety Population.
9.3.1.2.
Pharmacokinetic Concentration Population
The PK Concentration Population will include all subjects for whom at least one PK 
sample was obtained and analyzed.
9.3.1.3.
Pharmacokinetic Parameter Population
For each PK parameter, the PK Parameter Population will include all subjects who 
provide PK parameter data.
9.3.2.
Interim Analysis
No formal interim is planned. However, ongoing analyses of pharmacokinetic data will 
be performed in order to direct development of fixed dose formulations for subsequent 
study parts and the planned BE study.
9.4.
Key Elements of Analysis Plan
9.4.1.
Pharmacokinetic Analyses
Statistical analyses of the pharmacokinetic parameter data will be the responsibility of 
Clinical Statistics, GlaxoSmithKline. 
Pharmacokinetic data will be presented in graphical and/or tabular form and will be 
summarized descriptively by treatment. Descriptive summary table, graphics, and 
 
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treatment will also be provided. All pharmacokinetic data will be stored in the Archives, 
GlaxoSmithKline Pharmaceuticals, R&D. 
Following loge-transformation, AUC(0-∞), AUC(0-t) and Cmax of FDC formulations 
and reference will be separately analyzed using a mixed effects model with fixed effect 
terms for period and treatment. Subject will be treated as a random effect in the model. 
Point estimates and their associated 90% confidence intervals will be constructed for the 
differences, F1 – R, F2 – R, F3 – R, F4 – R for Part 1 and similarly for Part 2 (as needed); 
and X1-X2, Y1-Y2 for Part 3. The point estimates and their associated 90% confidence 
intervals will then be back-transformed to provide point estimates and 90% confidence 
intervals for the ratios, F1:R, F2-R:F3-R, F4-R; and X1:X2, Y1:Y2.
Tmax of FDC formulations will be analyzed with the non-parametric Wilcoxon Matched 
Pairs Method to compute point estimate and associated 90% confidence intervals for the 
median difference, F1 R, F2 R, F3 R, F4 R and X1:X2, Y1:Y2.
9.4.2.
Safety Analyses
Safety data will be presented in tabular and/or graphical format and summarized 
descriptively according to GSK’s CDISC standards.
10.
STUDY GOVERNANCE CONSIDERATIONS
10.1.
Posting of Information on Publicly Available Clinical Trial 
Registers
Study information from this protocol will be posted on publicly available clinical trial 
registers before enrollment of subjects begins.
10.2.
Regulatory and Ethical Considerations, Including the 
Informed Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.
The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent

Investigator reporting requirements (e.g. reporting of AEs/SAEs/protocol 
deviations to IRB/IEC)
 
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
GSK will provide full details of the above procedures, either verbally, in writing, 
or both.

Signed informed consent must be obtained for each subject prior to participation 
in the study

The IEC/IRB, and where applicable the regulatory authority, approve the clinical 
protocol and all optional assessments, including genetic research.  

Optional assessments (including those in a separate protocol and/or under separate 
informed consent) and the clinical protocol should be concurrently submitted for 
approval unless regulation requires separate submission.  

Approval of the optional assessments may occur after approval is granted for the 
clinical protocol where required by regulatory authorities.  In this situation, 
written approval of the clinical protocol should state that approval of optional 
assessments is being deferred and the study, with the exception of the optional 
assessments, can be initiated.
10.3.
Quality Control (Study Monitoring)

In accordance with applicable regulations including GCP, and GSK procedures, 
GSK monitors will contact the site prior to the start of the study to review with the 
site staff the protocol, study requirements, and their responsibilities to satisfy 
regulatory, ethical, and GSK requirements.  

When reviewing data collection procedures, the discussion will also include 
identification, agreement and documentation of data items for which the CRF will 
serve as the source document.
GSK will monitor the study and site activity to verify that the:

Data are authentic, accurate, and complete.

Safety and rights of subjects are being protected.

Study is conducted in accordance with the currently approved protocol and any 
other study agreements, GCP, and all applicable regulatory requirements.
The investigator and the head of the medical institution (where applicable) agrees to 
allow the monitor direct access to all relevant documents
10.4.
Quality Assurance

To ensure compliance with GCP and all applicable regulatory requirements, GSK 
may conduct a quality assurance assessment and/or audit of the site records, and 
the regulatory agencies may conduct a regulatory inspection at any time during or 
after completion of the study. 

In the event of an assessment, audit or inspection, the investigator (and institution) 
must agree to grant the advisor(s), auditor(s) and inspector(s) direct access to all 
relevant documents and to allocate their time and the time of their staff to discuss 
the conduct of the study, any findings/relevant issues and to implement any 
corrective and/or preventative actions to address any findings/issues identified.
 
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10.5.
Study and Site Closure

Upon completion or premature discontinuation of the study, the GSK monitor will 
conduct site closure activities with the investigator or site staff, as appropriate, in 
accordance with applicable regulations including GCP, and GSK Standard 
Operating Procedures.

GSK reserves the right to temporarily suspend or prematurely discontinue this 
study at any time for reasons including, but not limited to, safety or ethical issues 
or severe non-compliance. For multicenter studies, this can occur at one or more 
or at all sites.  

If GSK determines such action is needed, GSK will discuss the reasons for taking 
such action with the investigator or the head of the medical institution (where 
applicable).  When feasible, GSK will provide advance notification to the 
investigator or the head of the medical institution, where applicable, of the 
impending action.

If the study is suspended or prematurely discontinued for safety reasons, GSK will
promptly inform all investigators, heads of the medical institutions (where 
applicable) and/or institution(s) conducting the study. GSK will also promptly 
inform the relevant regulatory authorities of the suspension or premature 
discontinuation of the study and the reason(s) for the action.  

If required by applicable regulations, the investigator or the head of the medical 
institution (where applicable) must inform the IRB/IEC promptly and provide the 
reason for the suspension or premature discontinuation.
10.6.
Records Retention

Following closure of the study, the investigator or the head of the medical 
institution (where applicable) must maintain all site study records (except for 
those required by local regulations to be maintained elsewhere), in a safe and 
secure location.  

The records must be maintained to allow easy and timely retrieval, when needed 
(e.g., for a GSK audit or regulatory inspection) and must be available for review 
in conjunction with assessment of the facility, supporting systems, and relevant 
site staff.  

Where permitted by local laws/regulations or institutional policy, some or all of 
these records can be maintained in a format other than hard copy (e.g., microfiche, 
scanned, electronic); however, caution needs to be exercised before such action is 
taken.  

The investigator must ensure that all reproductions are legible and are a true and 
accurate copy of the original and meet accessibility and retrieval standards, 
including re-generating a hard copy, if required.  Furthermore, the investigator 
must ensure there is an acceptable back-up of these reproductions and that an 
acceptable quality control process exists for making these reproductions.
 
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
GSK will inform the investigator of the time period for retaining these records to 
comply with all applicable regulatory requirements.  The minimum retention time 
will meet the strictest standard applicable to that site for the study, as dictated by 
any institutional requirements or local laws or regulations, GSK 
standards/procedures, and/or institutional requirements. 

The investigator must notify GSK of any changes in the archival arrangements, 
including, but not limited to, archival at an off-site facility or transfer of 
ownership of the records in the event the investigator is no longer associated with
the site.
10.7.
Provision of Study Results to Investigators, Posting of 
Information on Publically Available Clinical Trials Registers 
and Publication
Where required by applicable regulatory requirements, an investigator signatory will be 
identified for the approval of the clinical study report.  The investigator will be provided 
reasonable access to statistical tables, figures, and relevant reports and will have the 
opportunity to review the complete study results at a GSK site or other mutually-
agreeable location.
GSK will also provide the investigator with the full summary of the study results.  The 
investigator is encouraged to share the summary results with the study subjects, as 
appropriate.
GSK will provide the investigator with the randomization codes for their site only after 
completion of the full statistical analysis.
The procedures and timing for public disclosure of the results summary and for 
development of a manuscript for publication will be in accordance with GSK Policy.
A manuscript will be progressed for publication in the scientific literature if the results 
provide important scientific or medical knowledge.
 
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11.
REFERENCES
Adcirca: EMA Summary of product Characteristics, Eli Lily, last updated 29-Sep-2015 at 
time of publication
EMEA guidelines on clinical development of fixed dose combination medicinal products. 
CHMP/EWP/240/95 Rev.1
Forgue S, Patterson B, Bedding A, et al. Tadalafil pharmacokinetics in healthy subjects. 
British Journal of Clinical Pharmacology. 20-Dec-2005
Galiè N et al. Initial Use of Ambrisentan plus Tadalafil in Pulmonary Arterial 
Hypertension. N Engl J Med 2015;373:834-44.
Galie` N, Humbert M, Vachiery J.L et al. 2015 ESC/ERS Guidelines for the diagnosis 
and treatment of pulmonary hypertension. European Heart Journal Advance Access, 
published August 29, 2015
Ghofrani AH, Galiè N, Grimminger F, et al. N Engl J Med 2013;369:330-40. Riociguat 
for the Treatment of Pulmonary Arterial Hypertension
Gilead Clinical Study report GS-US-300-0112: A Phase 1, Open-Label, Randomized, 
Two-Period, Crossover Study to Assess the Pharmacokinetics of Ambrisentan Alone and 
in the Presence of Multiple Doses of Tadalafil, and to assess the Pharmacokinetics of 
Tadalafil Alone and in the Presence of Multiple Doses of Ambrisentan in Healthy Adult 
Volunteers. Version 1.0, dated 27-June-2008
Gilead Investigator Brochure, Letaris, Volibris, Fifteenth edition: 06 April 2015
GSK Clinical Study Report for study AMB112565: AMBITION: A Randomized, 
Multicenter Study of First-Line Ambrisentan and Tadalafil Combination Therapy in 
Subjects with Pulmonary Arterial Hypertension. GSK document 2014N193963_00, dated 
13-Nov-14
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(a): 24 Table 3-2
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, editors. 
Contraceptive Technology. 19th edition. New York: Ardent Media, 2007(b): 28.
Liang F, Yang S, Yao L, Belardinelli L, Shryock J. Hypertension. 2012 Mar;59(3):705-
11. Ambrisentan and tadalafil synergistically relax endothelin-induced contraction of rat 
pulmonary arteries.
Pulido T, Adzerikho I, Channick RN, et al. N Engl J Med. 2013 Aug 29;369(9):809-18. 
Macitentan and Morbidity and Mortality in Pulmonary Arterial Hypertension
 
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Spence R, Mandagere A, Harrison B, et al.  J Pharm Sci. 2009 Dec;98(12):4962-74. No 
clinically relevant pharmacokinetic and safety interactions of ambrisentan in combination 
with tadalafil in healthy volunteers. 
Volibris: EMA Summary of Product Characteristics, GSK, variation II/41, at time of 
publication
 
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12.
APPENDICES
12.1.
Appendix 1: Abbreviations and Trademarks
Abbreviations
AE
Adverse Event
API
Active Pharmaceutical Ingredient
AUC
Area Under Curve
BE
Bioequivalence
CDISC
Clinical Data Interchange Standards Consortium
cGMP
Cyclic guanosine monophosphate
Cmax
Maximum concentration
CI
Confidence Interval
CV
Coefficient of variance 
CVw
Coefficient of variance within subject
EMA
European Medicines Agency 
ERA
Endothelin Receptor Antagonists
ERS
European Respiratory Society 
ESC
European Society of Cardiology
ET-1
Endothelin Receptor - 1
EU
European Union
FDA
Federal Drug Agency
FDC
Fixed Dose Combination
NO
Nitrous Oxide
PDE-5
Phosphodiesterase type 5
SAE
Serious Adverse Events
SRM
Study Reference Manual
tmax
Time to maximum concentration
t1/2
Half Life
WHO
World Health Organisation
Trademark Information
Trademarks of the GlaxoSmithKline 
group of companies
Trademarks not owned by the 
GlaxoSmithKline group of companies
NONE
Adcirca
WinNonlin
 
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12.2.
Appendix 2: Liver Safety Required Actions and Follow-Up
Assessments
Phase I Liver chemistry stopping criteria have been designed to assure subject safety and 
to evaluate liver event aetiology (in alignment with the FDA premarketing clinical liver 
safety guidance).
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf.
Phase I liver chemistry stopping criteria and required follow up assessments
Liver Chemistry Stopping Criteria – Liver Stopping Event
ALT-absolute
ALT≥3xULN
If  ALT≥3xULN  AND bilirubin1,2 2xULN (>35% direct bilirubin) or  INR >1.5, 
Report as an SAE.
See additional Actions and Follow-Up Assessments listed below
Required Actions and Follow up Assessments following Liver Stopping Event
Actions
Follow-Up Assessments

Immediately discontinue study treatment 

Report the event to GSK within 24 hours

Complete the liver event CRF, and complete 
an SAE data collection tool if the event also 
meets the criteria for an SAE2

Perform liver event follow up assessments 

Monitor the subject until liver chemistries 
resolve, stabilise, or return to within baseline 
(see MONITORING below)
MONITORING:
If ALT≥3xULN AND bilirubin 2xULN  or INR 
>1.5

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within  24 
hrs

Monitor subjects twice weekly until liver 
chemistries resolve, stabilise or return to 
within baseline

Viral hepatitis serology3

Blood sample for pharmacokinetic (PK) 
analysis, obtained within 96hrs of last dose

Serum creatine phosphokinase (CPK) and 
lactate dehydrogenase (LDH).

Fractionate bilirubin, if total 
bilirubin2xULN

Obtain complete blood count with 
differential to assess eosinophilia

Record the appearance or worsening of 
clinical symptoms of liver injury, or 
hypersensitivity, on the AE report form

Record use of concomitant medications on 
the concomitant medications report form 
including acetaminophen, herbal remedies, 
other over the counter medications.

Record alcohol use on the liver event 
alcohol intake case report form
If ALT≥3xULN AND bilirubin 2xULN  or 
 
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Liver Chemistry Stopping Criteria – Liver Stopping Event

A specialist or hepatology consultation is 
recommended
If ALT≥3xULN AND bilirubin < 2xULN  and 
INR ≤1.5:

Repeat liver chemistries (include ALT, AST, 
alkaline phosphatase, bilirubin) and perform 
liver event follow up assessments within 24-
72 hrs

Monitor subjects weekly until liver 
chemistries resolve, stabilize or return to 
within baseline
INR >1.5:

Anti-nuclear antibody, anti-smooth muscle 
antibody, Type 1 anti-liver kidney 
microsomal antibodies, and quantitative 
total immunoglobulin G (IgG or gamma 
globulins).

Serum acetaminophen adduct HPLC assay 
(quantifies potential acetaminophen 
contribution to liver injury in subjects with 
definite or likely acetaminophen use in the 
preceding week [James, 2009].

Liver imaging (ultrasound, magnetic 
resonance, or computerised tomography) 
and /or liver biopsy to evaluate liver 
disease; complete Liver Imaging and/or 
Liver Biopsy CRF forms.
1.
Serum bilirubin fractionation should be performed if testing is available. If serum bilirubin fractionation is not 
immediately available, discontinue study treatment for that subject if ALT 3xULN and bilirubin 2xULN. 
Additionally, if serum bilirubin fractionation testing is unavailable, record presence of detectable urinary bilirubin on 
dipstick, indicating direct bilirubin elevations and suggesting liver injury. 
2.
All events of ALT 3xULN and bilirubin 2xULN (>35% direct bilirubin) or ALT 3xULN and INR>1.5, if INR 
measured, which may indicate severe liver injury (possible ‘Hy’s Law’), must be reported as an SAE (excluding 
studies of hepatic impairment or cirrhosis); INR measurement is not required and the threshold value stated will 
not apply to subjects receiving anticoagulants
3.
Includes: Hepatitis A IgM antibody; Hepatitis B surface antigen and Hepatitis B Core Antibody (IgM); Hepatitis C
RNA; Cytomegalovirus IgM antibody;  Epstein-Barr viral capsid antigen IgM antibody (or if unavailable, obtain 
heterophile antibody or monospot testing);  Hepatitis E IgM antibody 
4.
PK sample may not be required for subjects known to be receiving placebo or non-GSK comparator treatments.  
Record the date/time of the PK blood sample draw and the date/time of the last dose of study treatment prior to 
blood sample draw on the CRF. If the date or time of the last dose is unclear, provide the subject’s best 
approximation. If the date/time of the last dose cannot be approximated OR a PK sample cannot be collected in 
the time period indicated above, do not obtain a PK sample. Instructions for sample handling and shipping are in 
the SRM. 
References
James LP, Letzig L, Simpson PM, Capparelli E, Roberts DW, Hinson JA, Davern TJ, Lee 
WM. Pharmacokinetics of Acetaminophen-Adduct in Adults with Acetaminophen 
Overdose and Acute Liver Failure. Drug Metab Dispos 2009; 37:1779-1784.
 
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12.3.
Appendix 3: Definition of and Procedures for Recording, 
Evaluating, Follow-Up and Reporting of Adverse Events
12.3.1.
Definition of Adverse Events
Adverse Event Definition:

An AE is any untoward medical occurrence in a patient or clinical investigation 
subject, temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product.

NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal product.
Events meeting AE definition include:

Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (e.g., ECGs, radiological scans, vital signs 
measurements), including those that worsen from baseline, and felt to be clinically 
significant in the medical and scientific judgement of the investigator.

Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

New conditions detected or diagnosed after study treatment administration even 
though it may have been present prior to the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected interaction.

Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication (overdose per se will not be reported as an 
AE/SAE unless this is an intentional overdose taken with possible suicidal/self-
harming intent.  This should be reported regardless of sequelae).
Events NOT meeting definition of an AE include:

Any clinically significant abnormal laboratory findings or other abnormal safety 
assessments which are associated with the underlying disease, unless judged by the 
investigator to be more severe than expected for the subject’s condition.

The disease/disorder being studied or expected progression, signs, or symptoms of 
the disease/disorder being studied, unless more severe than expected for the subject’s 
condition.

Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that 
leads to the procedure is an AE.

Situations where an untoward medical occurrence did not occur (social and/or 
 
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convenience admission to a hospital).

Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen.
12.3.2.
Definition of Serious Adverse Events
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, 
death due to progression of disease, etc).
Serious Adverse Event (SAE) is defined as any untoward medical occurrence that, 
at any dose:
a.
Results in death
b.
Is life-threatening
NOTE:
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
subject was at risk of death at the time of the event.  It does not refer to an event, which 
hypothetically might have caused death, if it were more severe.
c.
Requires hospitalization or prolongation of existing hospitalization
NOTE:

In general, hospitalization signifies that the subject has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for 
observation and/or treatment that would not have been appropriate in the physician’s 
office or out-patient setting.  Complications that occur during hospitalization are 
AEs.  If a complication prolongs hospitalization or fulfills any other serious criteria, 
the event is serious.  When in doubt as to whether “hospitalization” occurred or was 
necessary, the AE should be considered serious.

Hospitalization for elective treatment of a pre-existing condition that did not worsen 
from baseline is not considered an AE.
d.
Results in disability/incapacity
NOTE:

The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.

This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, 
and accidental trauma (e.g. sprained ankle) which may interfere or prevent everyday 
life functions but do not constitute a substantial disruption
 
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e.
Is a congenital anomaly/birth defect
f.
Other situations:

Medical or scientific judgment should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be 
immediately life-threatening or result in death or hospitalization but may jeopardize 
the subject or may require medical or surgical intervention to prevent one of the 
other outcomes listed in the above definition.  These should also be considered 
serious.

Examples of such events are invasive or malignant cancers, intensive treatment in an 
emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse
g.
Is associated with liver injury and impaired liver function defined as:

ALT 3xULN and total bilirubin* 2xULN (>35% direct), or

ALT 3xULN and INR** > 1.5.
* Serum bilirubin fractionation should be performed if testing is available; if unavailable, 
measure urinary bilirubin via dipstick.  If fractionation is unavailable and ALT 3xULN 
and total bilirubin 2xULN, then the event is still to be reported as an SAE.
** INR testing not required per protocol and the threshold value does not apply to 
subjects receiving anticoagulants.  If INR measurement is obtained, the value is to be 
recorded on the SAE form.

Refer to Section 12.2 for the required liver chemistry follow up instructions
12.3.3.
Definition of Cardiovascular Events
Cardiovascular Events (CV) Definition:
Investigators will be required to fill out the specific CV event page of the CRF for the 
following AEs and SAEs:

Myocardial infarction/unstable angina

Congestive heart failure

Arrhythmias

Valvulopathy

Pulmonary hypertension

Cerebrovascular events/stroke and transient ischemic attack

Peripheral arterial thromboembolism

Deep venous thrombosis/pulmonary embolism
 
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
Revascularization
12.3.4.
Recording of AEs and SAEs
AEs and SAE Recording:

When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) 
relative to the event.

The investigator will then record all relevant information regarding an AE/SAE in 
the CRF

It is not acceptable for the investigator to send photocopies of the subject’s medical 
records to GSK in lieu of completion of the GSK, AE/SAE CRF page.

There may be instances when copies of medical records for certain cases are 
requested by GSK.  In this instance, all subject identifiers, with the exception of the 
subject number, will be blinded on the copies of the medical records prior to 
submission of to GSK.

The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information.  In such cases, the diagnosis will be 
documented as the AE/SAE and not the individual signs/symptoms.

Subject-completed Value Evidence and Outcomes questionnaires and the collection 
of AE data are independent components of the study.

Responses to each question in the Value Evidence and Outcomes questionnaire will 
be treated in accordance with standard scoring and statistical procedures detailed by 
the scale’s developer.

The use of a single question from a multidimensional health survey to designate a 
cause-effect relationship to an AE is inappropriate.
12.3.5.
Evaluating AEs and SAEs
Assessment of Intensity
The investigator will make an assessment of intensity for each AE and SAE reported 
during the study and will assign it to one of the following categories:

Mild:  An event that is easily tolerated by the subject, causing minimal discomfort 
and not interfering with everyday activities.

Moderate:  An event that is sufficiently discomforting to interfere with normal 
everyday activities

Severe:  An event that prevents normal everyday activities. - an AE that is assessed 
as severe will not be confused with an SAE. Severity is a category utilized for rating 
 
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the intensity of an event; and both AEs and SAEs can be assessed as severe.

An event is defined as ‘serious’ when it meets at least one of the pre-defined 
outcomes as described in the definition of an SAE.
Assessment of Causality

The investigator is obligated to assess the relationship between study treatment and 
the occurrence of each AE/SAE.

A "reasonable possibility" is meant to convey that there are facts/evidence or 
arguments to suggest a causal relationship, rather than a relationship cannot be ruled 
out.

The investigator will use clinical judgment to determine the relationship.

Alternative causes, such as natural history of the underlying diseases, concomitant 
therapy, other risk factors, and the temporal relationship of the event to the study 
treatment will be considered and investigated.

The investigator will also consult the Investigator Brochure (IB) and/or Product 
Information, for marketed products, in the determination of his/her assessment.

For each AE/SAE the investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality.

There may be situations when an SAE has occurred and the investigator has minimal 
information to include in the initial report to GSK.  However, it is very important 
that the investigator always make an assessment of causality for every event 
prior to the initial transmission of the SAE data to GSK.

The investigator may change his/her opinion of causality in light of follow up 
information, amending the SAE data collection tool accordingly.

The causality assessment is one of the criteria used when determining regulatory 
reporting requirements.
Follow up of AEs and SAEs

The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as may be indicated or as requested by GSK to 
elucidate as fully as possible the nature and/or causality of the AE or SAE.

The investigator is obligated to assist.  This may include additional laboratory tests 
or investigations, histopathological examinations or consultation with other health 
care professionals.

If a subject dies during participation in the study or during a recognized follow up 
period, the investigator will provide GSK with a copy of any post-mortem findings, 
including histopathology.
 
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
New or updated information will be recorded in the originally completed CRF.

The investigator will submit any updated SAE data to GSK within the designated 
reporting time frames.
12.3.6.
Reporting of SAEs to GSK
SAE reporting to GSK via paper CRF

Facsimile transmission of the SAE paper CRF is the preferred method to transmit 
this information to the Medical Monitor or the SAE coordinator

In rare circumstances and in the absence of facsimile equipment, notification by 
telephone is acceptable, with a copy of the SAE data collection tool sent by 
overnight mail

Initial notification via the telephone does not replace the need for the investigator to 
complete and sign the SAE CRF pages within the designated reporting time frames.

Contacts for SAE receipt can be found at this beginning of the protocol on the 
Sponsor/Medical Monitor Contact Information page.
 
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12.4.
Appendix 4: High Fat Meal Content

2 eggs fried in butter,

2 strips of bacon,

2 slices of toast with butter,

120 g hash brown potatoes, and

240 mls of whole milk.
The standard high-fat meal will be the meal suggested by the US FDA in their 2002 draft 
guidance on food-effect bioavailability and bioequivalence studies. Approximately 50% 
of the caloric content of the meal is from fat and the meal is high in calories 
(approximately 1000 calories). Substitutions in this test meal can be made as long as the 
meal provides a similar amount of calories from protein, carbohydrate, and fat and has 
comparable meal volume and viscosity.
US Department of Health and Human Services.  Guidance for Industry. Food-Effect 
Bioavailability and Fed Bioequivalence Studies.  Guidance. Center for Drug Evaluation 
and Research (CDER). Food and Drug Administration; 2002.
 
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