A Phase I, First Time in Human, Two Part, Randomized, Placebo-Controlled, Double-Blind (Sponsor Unblind), Single and Repeat Dose Escalating Study to Evaluate the Safety, Tolerability and Pharmacokinetics of GSK3352589, a REarranged During Transfection (RET) Growth Factor Receptor Tyrosine Kinase Inhibitor, in Normal Healthy Volunteers
Phase: PHASE1
Conditions: Irritable Bowel Syndrome
Sponsor: GlaxoSmithKline

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TITLE PAGE
Protocol Title: A Phase I, First Time in Human, Two Part, Randomized, Placebo-
Controlled, Double-Blind (Sponsor Unblind), Single and Repeat Dose Escalating Study 
to Evaluate the Safety, Tolerability and Pharmacokinetics of GSK3352589, a REarranged 
during Transfection (RET) Growth Factor Receptor Tyrosine Kinase Inhibitor, in Normal 
Healthy Volunteers
Protocol Number: 207440
Short Title: A Phase 1, First Time in Human Study to Evaluate GSK3352589, a 
REarranged during Transfection (RET) Growth Factor Receptor Tyrosine Kinase 
Inhibitor, in Normal Healthy Volunteers
Compound Number:
GSK3352589
Sponsor Name and Legal Registered Address: 
GlaxoSmithKline Research & Development Limited
980 Great West Road
Brentford
Middlesex, TW8 9GS
UK
Medical Monitor Name and Contact Information: 
 MD, MS
Approval Date: 31-MAR-2017
Copyright 2017 the GlaxoSmithKline group of companies.  All rights reserved.  
Unauthorised copying or use of this information is prohibited.
 
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TABLE OF CONTENTS
PAGE
1.
SYNOPSIS...............................................................................................................6
2.
SCHEDULE OF ACTIVITIES (SOA).......................................................................10
3.
INTRODUCTION....................................................................................................19
3.1.
Study Rationale ..........................................................................................19
3.2.
Background ................................................................................................19
3.2.1.
Nonclinical ...................................................................................20
3.3.
Benefit/Risk Assessment ............................................................................22
3.3.1.
Risk Assessment .........................................................................22
3.3.2.
Benefit Assessment.....................................................................23
3.3.3.
Overall Benefit:Risk Conclusion...................................................23
4.
OBJECTIVES AND ENDPOINTS...........................................................................24
5.
STUDY DESIGN ....................................................................................................24
5.1.
Overall Design ............................................................................................24
5.2.
Number of Participants ...............................................................................25
5.2.1.
Part A ..........................................................................................25
5.2.2.
Part B ..........................................................................................26
5.3.
Participant and Study Completion...............................................................27
5.4.
Scientific Rationale for Study Design ..........................................................27
5.4.1.
Part A ..........................................................................................27
5.4.2.
Part B ..........................................................................................28
5.5.
Dose Justification........................................................................................28
5.5.1.
Human Pharmacodynamic Predictions ........................................28
5.5.2.
Human Pharmacokinetic Predictions ...........................................29
5.5.2.1.
Predicted exposures for higher than expected 
bioavailability..............................................................33
5.5.3.
Dose Range and Safety Margins .................................................33
6.
STUDY POPULATION...........................................................................................35
6.1.
Inclusion Criteria.........................................................................................35
6.2.
Exclusion Criteria........................................................................................36
6.3.
Lifestyle Restrictions...................................................................................37
6.3.1.
Part A Fasted:  Meals and Dietary Restrictions............................37
6.3.2.
Part A Fed for Pilot Food Effect (Cohort 2, Dosing Period 
2):  Meals and Dietary Restrictions ..............................................38
6.3.3.
Part B QD Fasted:  Meals and Dietary Restrictions......................38
6.3.4.
Part B BID Fasted:  Meals and Dietary Restrictions.....................39
6.3.5.
Part B QD Fed:  Meals and Dietary Restrictions ..........................40
6.3.6.
Part  B BID Fed:  Meals and Dietary Restrictions.........................40
6.3.7.
Caffeine, Alcohol, and Tobacco ...................................................41
6.3.8.
Activity .........................................................................................41
6.4.
Screen Failures...........................................................................................41
7.
TREATMENTS.......................................................................................................42
7.1.
Treatments Administered............................................................................42
 
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7.2.
Dose Modification .......................................................................................42
7.3.
Method of Treatment Assignment ...............................................................43
7.4.
Blinding.......................................................................................................43
7.5.
Preparation/Handling/Storage/Accountability..............................................43
7.6.
Treatment Compliance................................................................................44
7.7.
Concomitant Therapy..................................................................................44
7.8.
Treatment after the End of the Study ..........................................................45
8.
DISCONTINUATION CRITERIA.............................................................................45
8.1.
Discontinuation of Study Treatment ............................................................45
8.1.1.
Liver Chemistry Stopping Criteria ................................................45
8.1.2.
QTc Stopping Criteria ..................................................................46
8.2.
Withdrawal from the Study..........................................................................46
8.3.
Lost to Follow Up........................................................................................47
9.
STUDY ASSESSMENTS AND PROCEDURES .....................................................47
9.1.
Adverse Events...........................................................................................48
9.1.1.
Time Period and Frequency for Collecting AE and SAE 
Information...................................................................................48
9.1.2.
Method of Detecting AEs and SAEs.............................................48
9.1.3.
Follow-up of AEs and SAEs.........................................................49
9.1.4.
Regulatory Reporting Requirements for SAEs .............................49
9.1.5.
Pregnancy ...................................................................................49
9.2.
Treatment of Overdose...............................................................................49
9.3.
Safety Assessments ...................................................................................50
9.3.1.
Physical Examinations.................................................................50
9.3.2.
Vital Signs....................................................................................50
9.3.3.
Electrocardiograms......................................................................50
9.3.4.
Clinical Safety Laboratory Assessments ......................................51
9.3.5.
Bristol Stool Form Scale ..............................................................51
9.4.
Pharmacokinetics .......................................................................................51
9.4.1.
Sample Collection........................................................................51
9.4.1.1.
Blood sample collection..............................................51
9.4.1.2.
Urine Sample Collection .............................................52
9.4.2.
Sample Analysis ..........................................................................52
9.4.3.
Pharmacokinetic Analyses...........................................................52
9.5.
Pharmacodynamic Biomarkers ...................................................................53
9.6.
Genetics .....................................................................................................53
10. STATISTICAL CONSIDERATIONS........................................................................53
10.1.
Sample Size Determination ........................................................................53
10.2.
Populations for Analyses ............................................................................53
10.3.
Statistical Analyses.....................................................................................54
10.3.1.
Other Analyses ............................................................................54
10.3.2.
Interim Analyses ..........................................................................54
11. REFERENCES.......................................................................................................55
12. APPENDICES........................................................................................................57
12.1.
Appendix 1: Abbreviations and Trademarks................................................57
12.2.
Appendix 2: Human Pharmacodynamic Prediction Methods.......................60
 
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12.2.1.
Systems pharmacology model of human colon tissue 
concentrations .............................................................................60
12.2.2.
Human Equivalent Dose (HED) Prediction Method using 
FDA guidance with and without small and large intestine 
surface area correction ................................................................61
12.2.3.
Human Dose Prediction for of GSK3352589 Adjusted for 
Colon Content and Gut Weight ....................................................63
12.3.
Appendix 3: Clinical Laboratory Tests.........................................................64
12.4.
Appendix 4: Bristol Stool Form Scale..........................................................66
12.5.
Appendix 5: Study Governance Considerations..........................................67
12.6.
Appendix 6: Adverse Events: Definitions and Procedures for 
Recording, Evaluating, Follow-up, and Reporting .......................................71
12.7.
Appendix 7: Contraceptive Guidance and Collection of Pregnancy 
Information..................................................................................................76
12.8.
Appendix 8: Liver Safety: Required Actions and Follow-up 
Assessments ..............................................................................................80
 
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1.
SYNOPSIS
Protocol Title: A Phase I, First Time in Human, Two Part, Randomized, Placebo-
Controlled, Double-Blind (Sponsor Unblind), Single and Repeat Dose Escalating Study 
to Evaluate the Safety, Tolerability and Pharmacokinetics of GSK3352589, a REarranged 
during Transfection (RET) Growth Factor Receptor Tyrosine Kinase Inhibitor, in Normal 
Healthy Volunteers
Short Title: A Phase 1, First Time in Human Study to Evaluate GSK3352589, a 
REarranged during Transfection (RET) Growth Factor Receptor Tyrosine Kinase 
Inhibitor, in Normal Healthy Volunteers
Rationale:
Irritable bowel syndrome (IBS) is a relatively common gastrointestinal (GI) illness with a 
prevalence of approximately 5 to 20% and characterized by a constellation of clinical
symptoms including abdominal pain and discomfort, abnormal bowel habits, and bloating 
[Camilleri, 2012] [Halland, 2013] [Longstreth, 2006].  Because the etiology of the 
disease has not been established, diagnosis is difficult and relies primarily on the 
presence of a specific symptom complex occurring with a minimum frequency in the 
absence of an alternative explanation. It is generally believed that the sensory 
inputs/outputs in the peripheral and central nervous system are altered in such a way that 
a patient with IBS has a heightened and disproportionate sensory experience for a given 
stimulus, i.e. visceral hypersensitivity [Azpiroz, 2007].  Visceral hypersensitivity may 
result from increased nerve fiber density and sprouting in the intestinal mucosal tissues of 
patients with IBS [Dothel, 2015].
REarranged during Transfection (RET) is a neuronal growth factor receptor tyrosine 
kinase whose activity is critical for the development of the enteric nervous system (ENS), 
the kidney and spermatogenesis [Airaksinen, 2002].  While its role during the 
development of the ENS has been well established, recent data strongly implicate RET 
kinase in the maintenance and plasticity of the adult ENS. By reducing RET signaling, 
RET kinase inhibition may modulate sensory neuron activity in the enteric nervous 
system and ameliorate visceral hypersensitivity.  Preliminary in vitro and in vivo studies 
suggest that RET kinase inhibition is able to attenuate the function of cholinergic 
neurons. Taken together, these results suggest that treatment of IBS patients with a RET 
kinase inhibitor has the potential to reduce abdominal pain and discomfort in patients 
with IBS via a novel mechanism of action which may normalize both visceral 
hypersensitivity and abnormal motility by inhibiting the function of cholinergic neurons. 
Thus, the patient population that may derive the greatest benefit would consist of 
individuals with IBS-diarrhea (IBS-D).  
GSK3352589 is a potent and selective inhibitor of RET kinase.  The current study is 
designed to assess the safety, tolerability and PK of escalating single and repeat oral 
doses of GSK3352589 in normal healthy volunteers.  This study is the first administration 
of GSK3352589 to humans.
 
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Objectives and Endpoints:
Objective
Endpoint
Primary
Part A Single Dose

To evaluate the safety and tolerability of 
single escalating oral doses of 
GSK3352589 administered in the fasted 
state and a single dose of GSK3352589 
administered in the fed state in healthy 
adult subjects

To evaluate the pharmacokinetic 
parameters of escalating single oral 
doses of GSK3352589 under fasting and 
fed (for one dose only) conditions in 
healthy adult subjects
Part B Repeat Dose

To evaluate the safety and tolerability of 
repeat escalating oral doses of 
GSK3352589 administered once or twice 
daily in the fasted or fed state

To characterize the PK of GSK3352589 
of repeat escalating oral doses 
administered once or twice daily in the 
fasted or fed state

Clinical safety and tolerability data 
including spontaneous adverse event 
(AE) reporting, clinical observations, 
physical examination findings, 12-lead 
electrocardiograms (ECG), vital signs, 
Bristol Stool Form Scores (BSFS), clinical 
laboratory tests.  

PK parameters obtained following 
administration of GSK3353589.  PK 
parameters include areas under the 
plasma concentration versus time curve,
AUC over the dosing interval (and other 
time intervals), Cmax, Tmax, and terminal 
half life (t ½), for Days 1 and 14 will be 
analyzed as data permit.
Exploratory

To explore the effect of GSK3352589 on 
the postprandial plasma profiles of total 
glucagon like peptide-1 (GLP-1) and total 
peptide YY (PYY) following repeat oral 
doses of GSK3352589 administered once 
or twice daily 

Postprandial challenge (mixed-meal) 
profiles of total GLP-1 and total PYY.

To investigate the biotransformation of 
GSK3352589 in plasma and urine

Provide samples of plasma and urine for 
the identification of any compound-
derived metabolite(s).
 
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Overall Design:
This randomized, double-blind (sponsor unblinded), placebo controlled, dose escalation 
study will be divided into two parts: Part A is a single dose escalating, four period 
crossover (Cohort 1) with a two-period, single sequence pilot food effect study (Cohort 2) 
and Part B is a 14 day repeat dose escalating, ascending cohort study. 
The study will be composed of 3 periods for all subjects (Screening, Treatment, and 
Follow-up).  Subjects will participate in either Part A or Part B. A subject’s total time 
involved in the study will be approximately 10 weeks for subjects enrolled in Part A, 
Cohort 1 and approximately 8 weeks for subjects enrolled in Part A, Cohort 2.  Subjects 
enrolled in Part B will be involved in the study for a total time of approximately 7 weeks.
All dosing periods for subjects participating in Part A will be completed before beginning 
enrollment in Part B.  
Part A:
Number of Participants: A sufficient number of healthy volunteers (males and women 
of non-childbearing potential) will be screened to enroll 16 subjects (8 subjects/cohort).  
Treatment Period and Duration: The planned dose range for Cohort 1 is single doses 
of 2 to 400 mg in the fasted state.  The actual doses to be administered may be adjusted 
based on safety, tolerability and PK data at previous dose levels; these dose adjustments 
may involve either an increase or a decrease in the planned dose for Cohort 1. Dose 
escalation will occur only if mean systemic exposures are projected not to exceed the 
defined plasma toxicokinetic (TK) limits. A single dose of GSK33352589 will be 
administered in the fasted and fed states in Cohort 2.  The dose selected to evaluate the 
food effect will allow for a 2X increase in bioavailability and predicted exposures will 
not exceed the highest observed exposures at prior doses.
Cohort 1 (Single Dose Escalation1)
Treatment 
Sequence
Subjects
Period 1
Period 2
Period 3
Period 4
A
and
Placebo
D2
D3
D4
B
and
D1
Placebo
D3
D4
C
and
D1
D2
Placebo
D4
D
and
D1
D2
D3
Placebo
1
Dose escalation will occur only if mean systemic exposures are projected not to exceed the defined plasma 
toxicokinetic (TK) limits.
Cohort 2 (Pilot Food Effect)
Treatment Sequence
Subjects
Period 1
Period 2
E
D5
D5 + food
F
 and
Placebo
Placebo + food
 
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Part B: 
Number of Participants: Up to six (6) cohorts are planned and will be conducted 
sequentially.  A sufficient number of healthy volunteers (males and women of non-
childbearing potential) will be screened to randomize 48 subjects (8 subjects/cohort; 6 
subjects will be randomized to GSK3352589 and 2 subjects will be randomized to 
placebo).  
Treatment Period and Duration: The planned dose range is 2 mg once daily to 200 mg 
twice daily administered for 14 days. Selection of the starting dose for Part B, along with 
requirements for fasted or fed state study drug administration, will occur after review of 
the safety, tolerability and PK data of all subjects enrolled in Part A.  The actual doses to 
be administered may be adjusted based on safety, tolerability and PK data at previous 
dose levels; these dose adjustments may involve either an increase or a decrease in the 
planned dose. Dose escalation will occur only if mean systemic exposures are projected 
not to exceed the defined plasma toxicokinetic (TK) limits.  
 
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2.
SCHEDULE OF ACTIVITIES (SOA)
Part A: Cohort 1 and 2 – Single Dose – Screening and Day 1
Screening
Day -1
Day 1
Screen
(within 21d)
Day -11
Pre-
dose
0h
0.5h
1h
1.5h
2h
2.5h
3h
4h
5h
8h
12h
Informed Consent
X
Inclusion and exclusion criteria
X
Demography
X
Medical/medication/drug/alcohol 
history
X
X2
Full Physical Exam (Inc Wt, Ht)
X
X2
Vital Signs (BP, HR, Temperature)3
X
X
X
X
X
X
12-Lead ECG
X
X
X
X
X
Safety Labs (Chemistry, 
Hematology, and Urinalysis)
X
X
Pregnancy Test4
X
X
Estradiol/FSH5
X
Urine Drug Screen, Alcohol Screen & 
Cotinine6
X
X
HIV, Hep B, Hep C
X
Administration of mixed meal 
breakfast7
X
Dosing
X
 
 
 
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Screening
Day -1
Day 1
Screen
(within 21d)
Day -11
Pre-
dose
0h
0.5h
1h
1.5h
2h
2.5h
3h
4h
5h
8h
12h
Plasma PK Sampling8
X
X
X
X
X
X
X
X
X
X
X
Bristol Stool Form Scale (BSFS)9
============================================================
Concomitant Medication
=================================================
AE Assessment
===================================================================
SAE Assessment
========================================================================
Outpatient Visits
X
Inpatient Check-in
X
1.
Within each cohort, subjects will return for their next scheduled dosing period approximately 14 days after administration of the study drug during the prior dosing period
2.
Period 1 Day -1:  Update medical/ medication history; No Wt/ Ht at Day-1.
3.
Full vital signs to be taken at screening and Day-1, and on Day 1 predose.  Blood pressure and heart rate will be taken 1 hr, 4 hr and 12 hr after administration of study drug on 
Day 1.
4.
Serum hCG at screening, urine hCG pre-dose on Day -1 – all females.
5.
If indicated
6.
An alcohol breath test is acceptable.
7.
Mixed meal breakfast ONLY for Cohort 2, Period 2 immediately following dosing.
8.
PK will be collected on Day 1, 2 and 3 with time relative to Day 1 dose.  Day 1 at predose within 10 minutes prior to 1st dose at time zero and then postdose at the following 
timepoints: 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 8, 12 hrs. 
9.
BSFS for all stool samples beginning Day -1.  
 
 
 
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Part A: Cohort 1 and 2 – Single Dose – Days 2-3 and Follow Up
Day 2
Day 2
Day 2
Day 3
Follow Up1
16h
24h
36h
48h
(Days 7-10)
Medical/medication/drug/alcohol history
Brief Physical Exam 
X2
X2
Vital Signs (BP, HR, Temperature)
X
X
X
12-Lead ECG
X
X
Safety Labs (Chemistry, Hematology, and Urinalysis)
X
X
Pregnancy Test3
X
Plasma Pharmacokinetic Sampling4
X
X
X
X
BSFS5
===========================
Concomitant Medication
===========================
AE Assessment
===========================
SAE Assessment
===========================
Outpatient Visits
X
Inpatient Check-out
X
1.
Subjects will return approximately 1 week after check out from their last actual dosing period (i.e. Day 1 of Dosing Period 3) for a final follow up visit.  If a subject discontinues from 
the study prior to completing all 4 periods in Cohort 1 or all 2 periods in Cohort 2, they will complete their early termination visit 7-10 days after their last dose received.
2.
No Wt/ Ht, Brief, symptom directed Physical Exam.
3.
Serum hCG at follow up – all females.
4.
PK will be collected on Days 1, 2 and 3 with time relative to Day 1 dose.  Day 2 at: 16, 24, & 36 hrs & Day 3 at 48 hrs.
5.
BSFS for all stool samples beginning Day -1
 
 
 
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Part B – Repeat Dose – Fasted (QD & BID)
Screening 
Period and 
Baseline 
Assessments
(within 21 
days)
Treatment 
Period
Follow Up 
Period or 
Early 
Termination
Screening
Day -2
Day -1
Day 1
Days 
2-6
Day 7
Day 
8-12
Day 13
Day 14
Day 15
Day 22-25
Informed Consent
X
Inclusion and exclusion criteria
X
X1
Demography
X
Medical/surgical/alcohol history
X
X1
Full Physical Exam (Inc Wt, Ht)
X
X1
X1
X1
X1
Vital Signs (BP, HR, Temperature) 2
X
X
X
X
X
X
X
X
X
X
X
12-Lead ECG3
X
X
X
X
X
Safety Labs (Chemistry, Hematology, 
and Urinalysis)
X
X
X
X
X
Pregnancy Test4
X
X
X
Estradiol/FSH5
X
Cotinine Test, Urine Drug Screen & 
Alcohol Screen6
X
X
HIV, Hep B, Hep C
X
Administration of standard meal at 
dinner
X
X
X
Administration of mixed meal at 
breakfast7
X
X
X
X
 
 
 
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Screening 
Period and 
Baseline 
Assessments
(within 21 
days)
Treatment 
Period
Follow Up 
Period or 
Early 
Termination
Screening
Day -2
Day -1
Day 1
Days 2-
6
Day 7
Day 8-12
Day 13
Day 14
Day 15
Day 22-25
Pharmacodynamic testing8
X
X
X
Dosing9
X
X
X
X
X
X
Plasma PK Sampling10
X
X
X
X
X
X
Plasma Metabolite Sampling11
X
X
Urinary PK &Metabolite Sampling12
X
X
Bristol Stool Form Scale (BSFS)13
========================================
Prior and Concomitant Medication
================================================
AE Assessment
================================
SAE Assessment
==========================================================
Outpatient Visits
X
X
Inpatient Check-in
X
Inpatient Check-out
X
1.
Day -2: Confirm eligibility, update medical / medication history. Ht at Screening only.  Wt at screening and on Days -1, 7 and 15.  Brief symptom directed physical exam on Days 7, 
15 and at FU.
2.
Full vital signs will be taken at screening, D-2, D-1, Day 1, 7 and 14 predose, D15, & FU.  Vital signs on Day 1, 7 and 14 will be taken 4 hrs after admin of study drug.
3.
12 lead ECG will be on Days 1, 7 and 14 predose and approximately 4 hours after administration of study drug.
4.
Serum hCG at screening and follow up, urine hCG pre-dose on Day -2 – all females.
5.
If indicated 
6.
An alcohol breath test is acceptable.  
7.
Mixed meal breakfast to be eaten on days -1, 1, 7 and 14 at approximately 2 hours post dose and eaten within 15 minutes.
 
 
 
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8.
Blood samples for PD on Day -1, Day 1 and Day 14 at the following timepoints: 15, 10, and 5 minutes prior to breakfast, and 0.5, 1, 2, and 4 hrs post breakfast.  Lunch to be 
eaten after 4 hr sample.  
9.
QD Dosing will be at approximately 0800 hours. BID Dosing will be at approximately 0800 and PM dose to be administered 10 hours following AM dose.
10. For QD dosing: Days 1 and 14 PK blood samples will be obtained predose within 10 minutes prior to 1st dose at time zero and then postdose at the following timepoints: 0.5, 1, 
1.5, 2, 2.5, 3, 4, 5, 8, 12, 16, and 24 hrs (within 10 minutes or less prior to next dose).
For QD dosing, Day 7: PK blood samples will be obtained predose within 10 minutes prior to 1st dose at time zero and then postdose at the following timepoints: 1, 2, 4, 8, and 24
hours (10 minutes prior to next dose on Day 8).
For BID dosing: Days 1 and 14 PK blood samples will be obtained predose within 10 minutes prior to 1st dose and then postdose at the following timepoints: 0.5, 1, 1.5, 2, 2.5, 3, 
4, 5, 8, 10 (10 mins prior to 2nd dose), 11, 12, 14, 16, and 24 hrs (within 10 minutes or less prior to next dose).
For BID dosing, Day 7: PK blood samples will be obtained predose within 10 minutes prior to 1st dose at time zero and then postdose at the following timepoints: 1, 2, 4, 8, 10 
hours (10 minutes or less prior to 2nd dose) and 24 hours (10 minutes prior to next dose on Day 8).
11. Plasma samples for metabolite profiling will be collected pre-dose and at all PK timepoints post-dose (for 0-24 hours for all cohorts, however only samples from the highest dose 
cohort will be analyzed.
12. Urine samples to aid metabolite structure identification will be collected pre-dose and over 0-24 hours for all cohorts and analyzed for the highest dose cohort samples only
13. BSFS for all stool samples beginning with time of check into clinical facility.
 
 
 
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Part B – Repeat Dose – Fed (QD and BID)
Screening 
Period and 
Baseline 
Assessments
(within 21 
days)
Treatment 
Period
Follow Up 
Period or 
Early 
Termination
Screening
Day -2
Day -1
Day 1
Days 2-6
Day 7
Day 8-
12
Day 13
Day 14
Day 15
Day 22-25
Informed Consent
X
Inclusion and exclusion criteria
X
X1
Demography
X
Medical/surgical/alcohol history
X
X1
X1
X1
Full Physical Exam (Inc Wt, Ht)
X
X1
X1
X1
X1
Vital Signs (BP, HR, Temperature) 2
X
X
X
X
X
X
X
X
X
X
X
12-Lead ECG3
X
X
X
X
X
Safety Labs (Chemistry, Hematology, 
and Urinalysis)
X
X
X
X
X
Pregnancy Test4
X
X
X
Estradiol/FSH5
X
Cotinine Test, Urine Drug Screen & 
Alcohol Screen6
X
X
HIV, Hep B, Hep C
X
Administration of standard meal at 
dinner
X
X
X
Administration of mixed meal at 
breakfast7
X
X
X
X
 
 
 
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Screening 
Period and 
Baseline 
Assessments
(within 21 
days)
Treatment 
Period
Follow Up 
Period or 
Early 
Termination
Screening
Day -2
Day -1
Day 1
Days 2-6
Day 7
Day 8-
12
Day 13
Day 14
Day 15
Day 22-25
Pharmacodynamic testing8
X
X
X
Dosing9
X
X
X
X
X
X
Plasma PK Sampling10
X
X
X
X
X
X
Plasma Metabolite Sampling11
X
X
Urinary PK &Metabolite Sampling12
X
X
Bristol Stool Form Scale (BSFS)13
================================
Prior and Concomitant Medication
======================================================
AE Assessment
====================================
SAE Assessment
========================================================
Outpatient Visits
X
X
Inpatient Check-in
X
Inpatient Check-out
X
1.
Day - 2: Confirm eligibility, update medical / medication history. Ht at Screening only.  Wt at screening and on Days -1, 7 and 15.  Brief symptom directed physical exam on Days 
7, 15 and at FU.
2.
Full vital signs will be taken at screening, D-2, D-1, Day 1, 7 and 14 predose, D15, & FU.  Vital signs on Day 1, 7 and 14 will be taken 4 hrs after admin of study drug.
3.
12 lead ECG will be on Days 1, 7 and 14 predose and approximately 4 hours after administration of study drug.
4.
Serum hCG at screening and follow up, urine hCG pre-dose on Day -2 – all females.
5.
If indicated 
6.
An alcohol breath test is acceptable.
7.
Mixed meal breakfast to be eaten on days -1, 1, 7 and 14 immediately following dose (approx 0800) and eaten within 15 minutes.  
 
 
 
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8.
Blood samples for PD on Day -1, Day 1 and Day 14 at the following timepoints: 15, 10, and 5 minutes prior to breakfast and 0.5, 1, 2, and 4 hrs post breakfast.  Lunch to be 
eaten after 4 hr sample obtained
9.
QD Dosing will be at approximately 0800 hours and BID dosing will be at approximately 0800 and PM dose to be administered 10 hours following AM dose.
10. For QD dosing: Days 1 and 14 PK blood samples will be obtained predose within 10 minutes prior to 1st dose at time zero and then postdose at the following timepoints: 0.5,1, 
1.5, 2, 2.5, 3, 4, 5, 8, 12, 16, and 24 hr (within 10 minutes or less prior to next dose).
For QD dosing, Day 7: PK blood samples will be obtained predose within 10 minutes prior to 1st dose at time zero and then postdose at the following timepoints: 1, 2, 4, 8, 
and 24 hours (10 minutes prior to next dose on Day 8).
For BID dosing: Days 1 and 14 PK blood samples to be obtained predose within 10 minutes prior to 1st dose and then postdose at the following timepoints: 0.5, 1, 1.5, 2, 
2.5, 3, 4, 5, 8, 10 (10 mins prior to 2nd dose), 11, 12, 14, 16, and 24 hrs (within 10 minutes or less prior to next dose ). 
For BID dosing, Day 7: PK blood samples will be obtained predose within 10 minutes prior to 1st dose at time zero and then postdose at the following timepoints: 1, 2, 4, 8, 
10 (10 mins prior to 2nd dose) and 24 hrs (10 minutes or less prior to next dose).
11. Plasma samples for metabolite profiling will be collected pre-dose and at all PK timepoints post-dose (for 0-24 hours for all cohorts, however only samples from the highest dose 
cohort will be analyzed.
12. Urine samples to aid metabolite structure identification will be collected pre-dose and over 0-24 hours for all cohorts and analyzed for the highest dose cohort samples only.
13. BSFS for all stool samples beginning with time of check into clinical facility.
 
 
 
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3.
INTRODUCTION
Irritable bowel syndrome (IBS) is a relatively common gastrointestinal (GI) illness with a 
prevalence of approximately 5 to 20% and characterized by a constellation of clinical 
symptoms including abdominal pain and discomfort, abnormal bowel habits, and bloating 
[Camilleri, 2012] [Halland, 2013] [Longstreth, 2006].  Because the etiology of the 
disease has not been established, diagnosis is difficult and relies primarily on the 
presence of a specific symptom complex occurring with a minimum frequency in the 
absence of an alternative explanation. It is generally believed that the sensory 
inputs/outputs in the peripheral and central nervous system are altered in such a way that 
a patient with IBS has a heightened and disproportionate sensory experience for a given 
stimulus, i.e. visceral hypersensitivity [Azpiroz, 2007].  Visceral hypersensitivity may 
result from increased nerve fiber density and sprouting in the intestinal mucosal tissues of 
patients with IBS [Dothel, 2015].
The only drugs which are FDA-approved for treatment of IBS are alosetron, rifaximin 
and eluxadoline for refractory diarrhea-predominant IBS, and linaclotide and 
lubiprostone for constipation-predominant IBS.  The majority of these drugs and those 
used off label to treat IBS, treat patient symptoms by altering GI motility and are 
minimally effective in addressing abdominal pain and discomfort.  Of the approved 
agents, only alosetron is hypothesized to modulate the central and enteric nervous system. 
[Berman, 2002] [Gyermek, 1996]
3.1.
Study Rationale
The current study is designed to assess the safety, tolerability and PK of escalating single 
and repeat oral doses of GSK3352589 in normal healthy volunteers.  This study is the 
first administration of GSK3352589 to humans.
3.2.
Background
REarranged during Transfection (RET) is a neuronal growth factor receptor tyrosine 
kinase whose activity is critical for the development of the enteric nervous system, the 
kidney and spermatogenesis [Airaksinen, 2002].  The role of RET kinase in the 
development of the ENS has been aided by the study of patients with Hirschsprung’s 
disease who frequently suffer from colonic obstruction due to a lack of normal colonic 
innervation and possess RET loss of function mutations [Edery, 1994].  While its role 
during the development of the ENS has been well established, recent reports also 
implicate a significant role for RET kinase in the maintenance and plasticity of the adult 
ENS.  Indeed, neurons within the submucosal and myenteric plexus of the adult human 
colon have been shown to express RET kinase and its co-receptors glial cell line-derived 
neurotrophic factor family receptors alpha 1 and 2 (GFR1 and GFR2) while glial cell 
line-derived neurotrophic factor (GDNF), the ligand for RET kinase, is expressed in the 
muscularis mucosa, and circular and longitudinal muscle tissue [Barrenschee, 2013].  
Systemic administration of GDNF in adult rodents results in significant increases in 
submucosal neuron density in both the small intestine and colon and altered function 
[Wang, 2010].  Furthermore, a conditional knockout of the RET kinase co-receptor, 
GFRα3, results in decreased colonic hypersensitivity implicating a role for RET kinase 
 
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signaling in visceral nociception [Tanaka, 2011].  Therefore, by reducing RET signaling, 
RET kinase inhibition may modulate sensory neuron activity in the enteric nervous 
system. 
GSK3352589 is a potent and selective inhibitor of RET kinase which has been shown to 
reduce visceral hypersensitivity in an animal model of IBS and inhibit cholinergic-
induced increases in colonic motility. The results from these preclinical studies suggest 
that RET kinase inhibition has the potential to reduce abdominal pain and discomfort in 
patients with IBS via a novel mechanism of action which may normalize both visceral 
hypersensitivity and abnormal motility by inhibiting the function of cholinergic neurons.  
Thus, the patient population that may derive the greatest benefit would consist of 
individuals with IBS-diarrhea with increased GI motility (IBS-D). 
GSK3352589 is the second RET kinase inhibitor to be assessed in normal volunteers.  To 
date, the first RET kinase inhibitor, GSK3179106, has been administered to 48 healthy 
volunteers in two Phase I clinical studies.  In general, administration of GSK3179106 to 
normal volunteers has been well tolerated.  Overall, systemic exposures GSK3179106 
were low due to poor oral bioavailability. A significant food effect was observed; the 
geometric mean ratio comparing fed to fasted for AUC0-t and Cmax was approximately 10-
fold and 8-fold, respectively. No clinically relevant risks that would preclude dosing a 
RET kinase inhibitor for up to 2 weeks in normal healthy volunteers have been identified.
GSK3352589 has different physicochemical properties than GSK3179106.  GSK3352589 
is a Biopharmaceutical Classification System (BCS) Class 3 compound with high 
solubility and low permeability and is expected to have more homogeneous GI tract 
tissue distribution which has the potential for improved effectiveness. Additionally, 
GSK3352589 systemic exposures are expected to be lower when compared to 
GSK3179106 and are not expected to increase when administered with food.
3.2.1.
Nonclinical
GSK3352589 is a potent ATP non-competitive reversible inhibitor of the RET kinase 
enzyme in biochemical and cell-based assays with a mean IC50’s of 0.07 and 21 nM 
respectively.  In vivo, the effect of GSK3352589 was assessed in a rat model of irritable 
bowel syndrome (IBS) for the inhibition of acetic acid induced colonic hypersensitivity 
as measured by the visceromotor response to colonic distension at different pressures 
[Plourde, 1997].  GSK3352589 significantly inhibited rat colonic hypersensitivity when 
administered orally twice daily at 10 mg/kg/dose for 3.5 days when compared to vehicle 
controls (49 and 43% inhibition at 40 and 60 mmHg, respectively; P<0.0001). Similarly, 
doses of GSK3352589 at 30 and 100 mg/kg/dose inhibited colonic hypersensitivity 
compared to vehicle controls (30 mg/kg/dose: 43 and 40% inhibition; 100 mg/kg/dose: 45 
and 44% inhibition at 40 and 60 mmHg, respectively, P<0.0001). GSK3352589 at all 
doses examined had no effect on stool consistency or colonic motility in the basal state. 
In contrast, GSK3352589 was able to inhibit increased colonic motility stimulated with 
the use of the cholinergic agonist neostigmine at doses ≥ 3mg/kg/dose.
In addition to its expression in the enteric nervous system, RET kinase is expressed in 
enteroendocrine cells lining the intestinal mucosa (data on file).  Preliminary test results 
evaluating the effect of a tool compound that inhibits RET kinase suggest a potential role 
 
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of RET kinase in regulating levels of postprandial glucagon-like peptide-1 (GLP-1) (data 
on file).  Safety pharmacology studies have shown that GSK3352589 caused no overt 
treatment-related acute neurobehavioral effects in the conscious rat following single 
doses of 10, 100 or 1000 mg/kg.  GSK3352589 produced no treatment-related effects on 
the cardiovascular system in conscious dogs.  GSK3352589 had no effect on respiratory 
ventilation parameters in dogs after single doses of 10, 100 or 1000 mg/kg.  GSK3352589
produced a concentration-dependent prolongation in the QT interval in rabbit ventricular 
wedge preparations. Based on this finding, an investigative cardiovascular study was 
performed in conscious dogs.  Intravenous infusions of GSK3352589 yielding plasma 
concentrations greater than target concentrations determined by the rabbit ventricular 
wedge assay indicated no ECG waveform abnormalities/ arrhythmias or QT prolongation 
attributable to GSK3352589.  IC50 values for inhibition of hERG channel tail current 
could not be achieved at concentrations up to 44.67 M, however concentration-
dependent inhibition allowed calculation of an IC25 which was estimated to be 28.22 M 
(15.55 g/mL).  As these in vitro concentrations, which represent free drug, greatly 
exceed those anticipated in humans, it is considered unlikely that GSK3352589 would 
prolong QT interval at the proposed clinical dose.  In addition, as described above, no 
effects on the ECG measurements, including QT interval measurements, made at regular 
intervals throughout studies in which animals received GSK3352589 at doses up to 
1000 mg/kg/day for 28 days were observed.  It is therefore concluded that GSK3352589
is unlikely to cause any effects on the ECG in humans.
No test article-related findings were noted in the 1 month rat and dog oral toxicity 
studies, therefore the NOAEL was considered to be greater than or equal to 1000 
mg/kg/day, the highest dose tested. In dogs, the highest exposure was observed where 
gender averaged AUC(0-t) and Cmax were 40.4 ng.h/mL and 26.6 ng/mL, respectively; in 
rats gender averaged AUC(0-t) and Cmax were 51 ng.h/mL and 7.7 ng/mL, respectively.
In a non-GLP study conducted in rats, a dose range of GSK3352589 was given by 
intravenous infusion, 4 hours daily, for 7 consecutive days to identify target organs at 
systemic concentrations exceeding predicted clinical exposure.  Test article-related
effects were limited to cortical lymphocytolysis in the thymus and decreased lymphocyte 
and total white blood cell counts.  Systemic exposure in rats given 5 mg/kg/day by 4 hour 
daily intravenous infusion produced an AUC(0-t) and Cmax of 2490 ng.h/mL and 
578 ng/mL, respectively.
The genetic toxicity of GSK3352589 has been investigated in preliminary Ames and 
Mouse Lymphoma Assay (MLA) assays.  No mutagenic potential was demonstrated in 
either the absence or presence of S9-mix in these assay systems.
Low systemic exposure following single and repeat dose administration of the oral
formulations has been demonstrated in these nonclinical studies, consistent with very low 
oral bioavailability (rats2%; dogs1%). 
Preliminary data from two non-GLP dose ranging embryofetal development (EFD) 
studies conducted with GSK3352589 in rats and rabbits demonstrated no findings at 
exposures much higher than those achievable by oral administration (Cmax 623 ng/mL 
and AUC 5080 ng.h/mL).
 
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Detailed summaries of all completed in vitro and in vivo nonclinical safety pharmacology 
and toxicity studies are provided in the GSK33352589 Investigator’s Brochure 
[GlaxoSmithKline Document Number 2016N305051_00].
3.3.
Benefit/Risk Assessment
Summaries of findings from non-clinical studies conducted with GSK3352589 can be 
found in the Investigator’s Brochure [GlaxoSmithKline Document Number 
2016N305051_00].  The following section outlines the risk assessment and mitigation
strategy for this protocol.
3.3.1.
Risk Assessment
The current study 207440 represents the first administration of GSK3352589 to healthy 
subjects and extensive safety monitoring will occur during the course of the study.  
Considerations for safety monitoring are derived primarily from experience dosing the 
first RET kinase inhibitor (GSK3179106) to normal healthy volunteers and GSK3352589 
non-clinical data. Administration of GSK3179106 in single doses up to 800 mg QD in the 
fasted state and repeat doses up to 200 mg BID, in the fed state, was in general, well 
tolerated with no clinically important AEs observed that would preclude dosing of 
GSK3352589 to normal healthy volunteers for up to 14 days.  The safety and tolerability 
profile of GSK3179106 is attributed to its low oral bioavailability.
Based on the absence of test article related adverse findings in the GLP safety 
pharmacology and repeat dose toxicity studies, coupled with the likelihood of very low 
GSK3352589 systemic exposures highest planned AUC exposure = 40.4 ng.h/mL, the 
risk of adverse events occurring in healthy adult male volunteers and female volunteers 
of non childbearing potential administered single or repeat doses of GSK3352589 in the 
fasted or fed state is judged to be low.
More detailed information about the known and expected benefits and risks and 
reasonably expected adverse events of GSK3352589 may be found in the the 
Investigator’s Brochure [GlaxoSmithKline Document Number 2016N305051_00].
Potential Risk of Clinical
Significance
Summary of Data/Rationale 
for Risk
Mitigation Strategy
Investigational Product (IP) [GSK 3352589]
Embryofetal development 
toxicity
Potential class effect of 
tyrosine kinase inhibitors. 
Lead RET kinase inhibitor 
is a multi-species teratogen.
Early studies with 
GSK3352589 suggest that it 
is not teratogenic.  
Definitive studies are 
planned.
Women of child bearing 
potential excluded from 
study participation.  Men 
with female partners 
required to use barrier 
contraception and 
females required to use a 
GSK approved method 
of hormonal 
contraception (see 
 
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Potential Risk of Clinical
Significance
Summary of Data/Rationale 
for Risk
Mitigation Strategy
Section 12.7).
Drug drug interaction
Given possibility of 
CYP3A4 and p-
glycoprotein inhibition in 
gut
Appropriate 
inclusion/exclusion 
criteria.
Potential GI effects
Based on biological 
properties (putative)
Routine PV and BSFS to 
monitor changes in 
defecation pattern. 
3.3.2.
Benefit Assessment
Participants in the study are not expected to receive any direct medical benefits from their 
participation in the study.
3.3.3.
Overall Benefit:Risk Conclusion
Taking into account the measures taken to minimize risk to subjects participating in this 
study, the potential risks identified in association with GSK3352589 are justified by the 
eventual benefits that may be afforded to patients with IBS.
 
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4.
OBJECTIVES AND ENDPOINTS
Objectives
Endpoints
Primary
Part A Single Dose

To evaluate the safety and tolerability of 
single escalating oral doses of 
GSK3352589 administered in the fasted 
state and a single dose of GSK3352589 
administered in the fed state in healthy 
adult subjects

To evaluate the pharmacokinetic 
parameters of escalating single oral doses 
of GSK3352589 under fasting and fed (for 
one dose only) conditions in healthy adult 
subjects.
Part B Repeat Dose

To evaluate the safety and tolerability of 
repeat escalating oral doses of 
GSK3352589 administered once or twice 
daily in the fasted or fed state.

To characterize the PK of GSK3352589 of 
repeat  escalating oral doses administered 
once or twice daily in the fasted or fed 
state

Clinical safety and tolerability data 
including spontaneous adverse event (AE) 
reporting, clinical observations, physical 
examination findings, 12-lead 
electrocardiograms (ECG), vital signs, 
Bristol Stool Form Scores (BSFS), clinical 
laboratory tests.  

PK parameters obtained following 
administration of GSK3353589.  PK 
parameters include areas under the 
plasma concentration versus time curve,
AUC over the dosing interval and other 
time intervals, Cmax, Tmax, and terminal 
t ½, for Days 1 and 14 will be analyzed as 
data permit.
Exploratory

To explore the effect of GSK3352589 on 
the postprandial plasma profiles of total 
glucagon like peptide-1 (GLP-1) and total 
peptide YY (PYY) following repeat oral 
doses of GSK3352589 administered once 
or twice daily

To investigate the biotransformation of 
GSK3352589 in plasma and urine

Postprandial challenge (mixed-meal) 
profiles of total GLP-1 and total PYY.

Provide samples of plasma and urine for 
the identification of any compound-derived 
metabolite(s).
5.
STUDY DESIGN
5.1.
Overall Design
This randomized, double-blind (sponsor unblinded), placebo controlled, dose escalation 
study will be divided into two parts: Part A is a single dose escalating, four period 
 
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crossover (Cohort 1) with a two period, single sequence pilot food effect (Cohort 2) study 
and Part B is a 14 day repeat dose escalating, ascending cohort study. 
The study will be composed of 3 periods for all subjects (Screening, Treatment, and 
Follow-up).  Subjects will participate in either Part A or Part B. A subject’s total time 
involved in the study will be approximately 10 weeks for subjects enrolled in Part A, 
Cohort 1 and approximately 8 weeks for subjects enrolled in Part A, Cohort 2.  Subjects 
enrolled in Part B will be involved in the study for a total time of approximately 7 weeks.
All dosing periods for subjects participating in Part A will be completed before beginning 
enrollment in Part B.  
5.2.
Number of Participants
5.2.1.
Part A
Single dose, dose escalating, four period, crossover (Cohort 1) with a two period, single 
sequence pilot food effect (Cohort 2) study.  
A sufficient number of healthy volunteers will be screened to enroll 16 subjects (8 
subjects/cohort).  Two cohorts of subjects are planned.  Subjects assigned to Cohort 1 
will participate in 4 dosing periods: 1 placebo and 3 dose escalating periods.  The planned 
dose range for Cohort 1 is 2 to 400 mg.  Each dosing period during Cohort 1 will be 
staggered so that only 2 of the 8 subjects will be administered study drug initially.  Once 
24 hours have elapsed, and provided there are no safety concerns, the remainder of 
subjects scheduled for that dosing period may be dosed.  A review of safety and 
tolerability will occur prior to administration of the next dose level.  This same procedure 
will be followed for each escalating dosing period.  Within each cohort, subjects will 
return for their next scheduled dosing period approximately 14 days after administration 
of the study drug during the prior dosing period.  
Cohort 2 will proceed after completion of the treatment periods in Cohort 1.  Subjects 
assigned to Cohort 2 will participate in a pilot food effect in Periods 1 and 2 (see Table 1
below).  A single dose of GSK3352589 will be administered in the fasted and fed states 
in Cohort 2. The dose selected to evaluate the food effect will allow for a 2X increase in 
bioavailability and predicted exposures will not exceed the highest observed exposures at 
prior doses.
The actual doses to be administered may be adjusted based on safety, tolerability and PK 
data at previous dose levels; these dose adjustments may involve either an increase or a 
decrease in the planned dose for Cohort 1. Dose escalation will occur only if mean 
systemic exposures are projected not to exceed the defined plasma toxicokinetic (TK) 
limits.  Preliminary safety and PK data from all subjects will be reviewed prior to the 
next Dosing Period.  The decision to proceed to the next dose level will be made by the 
GSK Medical Monitor and the investigator based on safety, tolerability and available PK 
data.  
 
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Table 1
Part A Treatment Sequence
Cohort 1 (Single Dose Escalation1)
Treatment 
Sequence
Subjects
Period 1
Period 2
Period 3
Period 4
A
and
Placebo
D2
D3
D4
B
and
D1
Placebo
D3
D4
C
and
D1
D2
Placebo
D4
D
and
D1
D2
D3
Placebo
1
Dose escalation will occur only if mean systemic exposures are projected not to exceed the defined plasma 
toxicokinetic (TK) limits.
Cohort 2 (Pilot Food Effect)
Treatment Sequence
Subjects
Period 1
Period 2
E
D5
D5 + food
F
and
Placebo
Placebo + food
5.2.2.
Part B
Part B is a 14 day randomized, double-blind, placebo-controlled, repeat dose escalating, 
ascending cohort study.  
Up to six (6) cohorts are planned and will be conducted sequentially.  A sufficient 
number of healthy volunteers will be screened to randomize 48 subjects (8 subjects/
cohort).  If subjects are prematurely discontinued from the study, additional replacement 
subjects may be enrolled at the discretion of the Sponsor in consultation with the 
Investigator.
The planned dose range is 2 mg once daily to 200 mg twice daily.  The starting dose for 
Part B will be selected after review of the safety, tolerability and PK data of subjects 
enrolled in Part A and completing all dosing periods.  An example dose escalation plan is 
shown in Table 2 below. The actual doses to be administered may be adjusted based on 
safety, tolerability and PK data at previous dose levels; these dose adjustments may 
involve either an increase or a decrease in the planned dose. Dose escalation will occur 
only if mean systemic exposures are projected not to exceed the defined plasma 
toxicokinetic (TK) limits.  
Table 2
Planned Dose Escalation for Part B
Regimen Descriptions
Number of Subjects
Cohort 1
2 mg of GSK3352589 administered QD
or matching placebo administered QD
GSK3352589 n=6
Placebo n=2
Cohort 2
10 mg of GSK3352589 administered QD
or matching placebo administered QD
GSK3352589 n=6
Placebo n=2
Cohort 3
25 mg of GSK3352589 administered BID
or matching placebo administered BID
GSK3352589 n=6
Placebo n=2
 
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Regimen Descriptions
Number of Subjects
Cohort 4
50 mg of GSK3352589 administered BID
or matching placebo administered BID
GSK3352589 n=6 
Placebo n=2
Cohort 5
100 mg of GSK3352589 administered BID
or matching placebo administered BID
GSK3352589 n=6
Placebo n=2
Cohort 6
200 mg of GSK3352589 administered BID
Or matching placebo administered BID
GSK3352589 n=6
Placebo n=2
Subjects will undergo pretreatment baseline pharmacodynamic (PD) testing after 
administration of a breakfast mixed-meal challenge on Day -1 prior to receiving the first 
dose of study drug, on Day 1, and at the end of the treatment period on Day 14.  
Preliminary safety and tolerability from 14 days of dosing and PK data from 7 days of 
dosing for each cohort will be reviewed prior to dose escalation and will be used to 
determine the dose to be administered in the subsequent cohort.  The decision to proceed 
to the next dose level will be made by the GSK Medical Monitor and the investigator 
based on safety, tolerability and available PK data.
Additional subjects/cohorts may be enrolled to allow for evaluation of additional dose 
levels or dosing regimens.
5.3.
Participant and Study Completion
A participant is considered to have completed the study if he/she has completed all 
phases of the study including the follow up visit
The end of the study is defined as the date of the last visit of the last participant in the 
study.
5.4.
Scientific Rationale for Study Design
5.4.1.
Part A
A randomized, placebo-controlled, double-blind (sponsor unblind), single ascending dose 
crossover study design is based on well established and published methods to evaluate 
the single dose administration of experimental drugs.  The use of this design reduces 
inter-subject variability in the PK data, thus improving its quality and interpretation.  
Additionally, it reduces the number of subjects exposed to the investigational agent and 
required to guide selection of the starting dose for the repeat dose study to follow.  
Approximately a 2 week washout period between administrations of escalating doses 
assures that study drug has been cleared and in the event that an adverse event occurs, 
adequate time for resolution.  Placebo has been selected as the control.
A pilot food effect study is included because of the potential for a food effect with a high 
solubility, low permeability compound such as GSK3352589.  Theoretical potential 
exists for a decrease in bioavailability that could limit tissue exposure.
Placebo has been selected as the control.
 
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5.4.2.
Part B
A randomized, placebo-controlled, double-blind (sponsor unblind), ascending cohort 
study design is based on well established and published methods to evaluate the repeat 
dose administration of experimental drugs.  Placebo has been selected as the control.
5.5.
Dose Justification
In the preclinical safety and repeat dose toxicity studies performed to date, GSK3352589
1000 mg/kg administered for up to 28 days by the oral route of administration, results in 
low exposure in rats or dogs without evidence of a test article related safety signal or 
finding.  Thus, the starting dose of 2 mg is derived from the minimum anticipated 
biological effect level (MABEL) dose related to its pharmacology rather than to NOAELs 
in the absence of safety or toxicity findings in animals [CDER, 2005].
5.5.1.
Human Pharmacodynamic Predictions
Five methods were used to estimate a therapeutic dose range for GSK3352589. The target 
tissue is the ENS within the colon. The pharmacological activity in the rat IBS visceral 
hypersensitivity model is driven by drug penetration from colon luminal contents to the 
ENS within colon tissue. The pharmacological response correlates with colon tissue 
concentrations rather than plasma concentrations. 
Because the pharmacology is thought to be driven by colon tissue drug concentrations, 
the human dose predictions are independent of the predicted human systemic 
pharmacokinetic profile.  A summary of the results for the various methods for 
translating rat to human pharmacology and doses are shown in Table 3. The dose range 
for minimal pharmacological effect is predicted to be 5 mg to 136 mg once daily. The 
dose range for maximum pharmacological effect is predicted to be 5 mg to 454 mg once 
daily.  It is unknown if these doses are best given once or twice daily, but twice daily 
dosing should provide more sustained tissue colon concentrations per day. The rat IBS 
visceral hypersensitivity model indicates steady state tissue concentrations and twice 
daily dosing is required.  Therefore low, single doses in humans are not expected to have 
an effect.  Detailed explanations of the methods are shown in Section 12.2.
 
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Table 3
Summary of human dose predictions by method
Method
Pharmacologically Active Doses
Minimum Effect
Maximum Effect
Rat IBS model dose reference
3 mg/kg
10 mg/kg
Systems pharmacology model
5-10 mg QD or >2 mg BID
HED - body surface area extrapolation
30 mg
100 mg
HED with relative small intestine surface 
area correction (rat to human)
7 mg
23 mg
HED with relative large intestine surface 
area correction (rat to human)
136 mg
454 mg
Human dose scaled by colon content and gut 
weight
122 mg
406 mg
HED = human equivalent dose (FDA- CDER 2005)
5.5.2.
Human Pharmacokinetic Predictions
The model PK parameters that relate dose and plasma GSK3352589 concentrations were
based on several interspecies scaling and prediction methods. Small molecule drug 
candidates with moderately high hepatic clearance and any size volumes of distribution 
are more reliably scaled between species based on body weights and a few other 
methods.  Absolute bioavailability and absorption rates are estimated more directly from 
the animal data.  The absolute bioavailability of GSK3353589 is expected to be very low 
and therefore the primary source of uncertainty for the predictions of concentration-time 
profiles and exposure estimates (i.e., AUC, Cmax).
The systemic clearance (CLs) was obtained from five interspecies scaling methods. The 
CLs median (range) was 836 mL/min (528-1002 mL/min). The magnitude of the CLs
estimate implies GSK3352589 is a moderately high hepatic clearance compound. The
median volume of distribution (V) was 314 liters ranging from 126 to 546 liters and 
implies drug distribution is very large and extensively distributed into tissues. 
Consequently, the median (range) for plasma half-life (T1/2) is 3.7 (2.1-7.9) hours. The 
Tmax, time to reach Cmax, is expected to be about 1.5 hours ranging from 0.5 to 5.0 
hours. The variability in all parameters was taken into account with simulations that 
include probability distributions for each parameter in the PK model equation describing 
the GSK3352589 concentration time profiles.
 
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In all nonclinical species tested, GSK3352589 has highly variable, limited, and less than 
proportional systemic exposure with increasing doses, consistent with its very low oral 
bioavailability (rats ≤2%; dogs ≤1%).  In the rat, oral administration with food slightly 
increased the systemic exposure of GSK3352589. The bioavailability (F) estimates used 
in the simulations are listed from low to high: 0.016% (mice solution, fasted), 0.02% 
(male rats GLP TK suspension, fed), 0.06% (rats suspension, fed), 0.07% (female rats 
GLP TK suspension, fed), 0.18% (rats solution, fasted), 0.24% (dogs solution, fed), 1.1% 
(rats non GLP TK suspension, fed), 1.9% (rats non GLP TK suspension, fed).  The F 
ranges from 0.016% to 1.9% and attributed to large variation in individual rats.  The 
overall group median is 0.125%.  More weight was given to the estimates of F that 
included all species and the predicted AUC and Cmax exposures for administration of 
single doses based on most likely bioavailability using F=0.16% can be seen in the Table 
4 below.  The higher estimates of 1.1% and 1.9% in rats appeared to be the outliers 
compared to other rat estimates, mice and dogs and the impact of assuming the highest 
potential exposure (1.5%) is shown in Table 6. These predictions were assumed to be 
linearly related to dose.  This assumption is conservative since animal PK shows either 
linear or less than dose proportional relationships. Plasma GSK3352589 concentration 
time profiles are shown in Figure 1 for two single doses (2 mg and 200 mg). The 
predicted AUC and Cmax exposures for repeat doses at steady state are shown in Table 5. 
The bioavailability of GSK3352589B is likely permeability-limited since it has low
permeability and high FaSSIF solubility and considered a biopharmaceutical 
classification system (BCS) Class III type compound.  Therefore, the compound is 
expected to be solubilized and well mixed in the gut contents and absorption across the 
enterocyte epithelial layer is the rate limiting step.  MALDI imaging studies indicate 
good GI tissue distribution despite a low permeability categorization.  With BCS class III 
compounds, it is more likely that there will be a decrease in bioavailability, where plasma 
AUC and Cmax are decreased [Radwan, 2012].  However, a food effect study in rats with 
rat chow showed an increase in plasma AUC of 2.6 fold.  The mixed prediction of a food 
effect will be considered when dose selection is made for the pilot food effect study.
Table 4
Predicted exposures based on mostly likely bioavailability estimates 
for single doses - Median (5th and 95th percentiles) predictions for 
plasma GSK3352589 AUC(0- ) and Cmax (FASTED) by dose
Dose (mg)
AUC(0-)a (h*ng/mL)
Cmaxa (ng/mL)
1
0.021 (0.0078, 0.095)
0.0028 (0.0013, 0.011)
2
0.044 (0.015, 0.19)
0.0058 (0.0026, 0.022)
5
0.105 (0.039, 0.48)
0.015 (0.0065, 0.06)
10
0.211 (0.077, 0.95)
0.029 (0.013, 0.11)
25
0.55 (0.19, 2.4)
0.075 (0.033, 0.29)
50
1.09 (0.39, 4.8)
0.15 (0.065, 0.57)
100
2.18 (0.77, 9.5)
0.29 (0.129, 1.14)
200
4.36 (1.53, 19.0)
0.579 (0.258, 2.27)
400
8.72 (3.06, 38.0)
1.16 (0.52, 4.54)
 
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a.
median (5th, 95th percentiles), n=10,000 simulations,
Additional Output:  median (range) Tmax (h) = 1.5 (0.5-5.0);  median [(5th, 95th percentiles) and range] T1/2(h) = 3.7 
[(2.1, 7.9) and 1.6-12],
Predictions assume dose proportionality and this assumption is generally consistent with animal PK though with 
highly variability,
Bioavailability (F) average 0.16% ranging from 0.016%-1.5%; F is built into V/F as input variable, the 5th, 95th
percentiles include the low and high F estimates,
Input: median (5th, 95th percentiles) for K01(h-1) = 1.7 (0.66, 3.9); K10 (h-1) = 0.19 (0.09, 0.33); Vd/F (Liters) = 
262,858 (67,595- 568,451); mean and range for Vd (Liters) = 314 (126-546),
Single dose human exposures AUC(0-) are equivalent to steady state AUC(0-24 h) with once daily dosing.
Table 5
Predicted exposures based on mostly likely bioavailability estimates 
for repeat doses at steady state - Median (5th and 95th percentiles) 
predictions for plasma GSK3352589 AUCss (FASTED) by dose
Once Daily Dosing
Twice Daily Dosing
Dose (mg)
AUCss(0-24) (h*ng/mL) a
AUCss(0-24) (h*ng/mL) a
1
0.021 (0.0078, 0.095)
0.042 (0.016, 0.19)
2
0.044 (0.015, 0.19)
0.088 (0.030, 0.38)
5
0.105 (0.039, 0.48)
0.210 (0.078, 0.96)
10
0.211 (0.077, 0.95)
0.422 (0.154, 1.90)
25
0.55 (0.19, 2.4)
1.10 (0.38, 4.80)
50
1.09 (0.39, 4.8)
2.18 (0.78, 9.6)
100
2.18 (0.77, 9.5)
4.36 (1.54, 19.0)
200
4.36 (1.53, 19.0)
8.72 (3.1, 38.0)
400
8.72 (3.06, 38.0)
-
a.
median (5th, 95th percentiles), n=10,000 simulations.
Predictions assume dose proportionality and this assumption is generally consistent with animal PK though highly 
variability.
Single dose human exposures AUC(0-) are equivalent to steady state AUCss(0-24 h) with once daily dosing 
assuming no time dependent changes in drug disposition.
 
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Figure 1
Simulation of median (5th, 95th percentiles) concentration-time
profiles in humans for 2 mg and 200 mg single doses of 
GSK3352589 assuming most likely bioavailability
2 mg 
Time (hour)
0
5
10
15
20
25
30
GSK3352589 plasma concentration (ng/mL)
0.000
0.005
0.010
0.015
0.020
0.025
0.030
median
5th and 95th percentile
200 mg
Time (hour)
0
5
10
15
20
25
30
GSK3352589 plasma concentration (ng/mL)
0.0
0.5
1.0
1.5
2.0
2.5
3.0
median
5th and 95th percentiles
 
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5.5.2.1.
Predicted exposures for higher than expected bioavailability
The PK parameter that is most uncertain is oral bioavailability (F).  Although exposures 
remain very low, results of preclinical PK and TK assessments suggest the potential for 
up to a 100X increase in bioavailability.  An analysis of predicted exposures for the 
highest potential oral bioavailability, 1.5% (gender averaged) is included to illustrate the
effect of bioavailability on AUC and Cmax for single doses in Table 6 below.  All other 
parameters predictions are the same as in Table 5 above.
Table 6
Predicted exposures based on highest possible bioavailability 
estimates for single doses - Median (5th and 95th percentiles) 
predictions for plasma GSK3352589 AUC(0- ) and Cmax (FASTED) 
by dose
Dose (mg)
AUC(0-)a (h*ng/mL)
Cmaxa (ng/mL)
1
0.26 (0.128, 0.61)
0.036 (0.024, 0.06)
2
0.52 (0.26, 1.22)
0.072 (0.048, 0.12)
5
1.30 (0.64, 3.04)
0.18 (0.12, 0.30)
10
2.60 (1.27, 6.07)
0.36 (0.23, 0.59)
25
6.5 (3.2, 15.2)
0.90 (0.60, 1.5)
50
13.0 (6.4, 30.3)
1.8 (1.2, 3.0)
100
26.0 (12.8, 60.5)
3.6 (2.3, 6.0)
200
52.0 (25.5, 121)
7.11 (4.61, 11.9)
400
104 (51, 242)
14.2 (9.2, 24)
a.
 median (5th, 95th percentiles), n=10,000 simulations,
Additional Output:  median (range) Tmax (h) = 1.5 (0.5-5.0);  median [(5th, 95th percentiles) and range] T1/2(h) = 3.7 
[(2.1, 7.9) and 1.6-12],
Predictions assume dose proportionality and this assumption is generally consistent with animal PK though with 
highly variability,
Bioavailability (F) = 1.5% for outlier simulations.
Input: median (5th, 95th percentiles) for K01(h-1) = 1.7 (0.66, 3.9); K10 (h-1) = 0.19 (0.09, 0.33); Vd/F (Liters) = 
21,353 (13,131- 30332); mean and range for Vd (Liters) = 314 (126-546),
Single dose human exposures AUC(0-) are equivalent to steady state AUC(0-24 h) with once daily dosing.
5.5.3.
Dose Range and Safety Margins
The MABEL for this therapeutic is related to its pharmacology rather than to any safety 
or toxicity findings.  The lowest doses predicted to be minimally effective following 
repeat dosing to steady-steady are 5 mg QD or 2 mg BID.  The proposed starting dose for 
this single dose study is 2 mg, a conservative estimate for minimal, if any, effect.  The 
highest doses predicted to be maximally effective following repeat dosing to steady state 
are 406 – 454 once daily. A highest total daily dose of 400 mg was selected based on the 
predictions for the maximally effective dose.
Table 7 provides the predicted systemic exposures for administration of single doses up 
to 400 mg of GSK3352589, assuming linearity for both the most likely estimate for 
bioavailability (0.16%) and the highest expected estimate for bioavailability (1.5%), and
the estimated safety margins for these predicted exposures, respectively.  The safety 
 
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margins are calculated from the systemic exposures observed in the 4 wk oral dog repeat 
dose toxicity study.  No test article-related findings were noted in the 1 month rat and dog 
oral toxicity studies, therefore the NOAEL was considered to be greater than or equal to 
1000 mg/kg/day, the highest dose tested in each species. In rats, the highest gender-
averaged exposures observed were AUC(0-t) and Cmax 51.0 ng.h/mL and 7.7 ng/mL, 
respectively; and in dogs the gender-averaged AUC(0-t)) and Cmax were 40.4 ng.h/mL 
and 26.7 ng/mL, respectively. Although these exposures are similar, the dog exposures 
were used to calculate the safety margins because of the higher Cmax.  
The safety margins for Cmax and AUC for the planned starting single dose of 2 mg 
GSK3352589 assuming the highest possible bioavailability (1.5%) are 742X and 155X, 
respectively.  The predicted mean steady-state Cmax and AUC for humans assuming 
dose proportionality administered the highest planned single dose of 400 mg are below 
the animal safety exposure limits if the exposures suggest that bioavailability is closer to 
the most likely predicted value, 0.16%. During both Part A (single dose) and Part B 
(repeat dose), dose escalation will occur only if mean systemic exposures are projected 
not to exceed the defined plasma toxicokinetic (TK) limits [AUC(0-t)) and Cmax of 40.4 
ng.h/mL and 26.7 ng/mL, respectively].  The risk of either exceeding the defined TK 
limits or occurrence of systemic adverse events is judged to be low because of the low 
oral bioavailability of GSK3352589.
Table 7
Predicted Most Likely and Highest Potential Human Systemic 
Exposures and Estimated Safety Margins for Single Doses of 
GSK3352589 (Fasted State)
Predicted Human 
Mean Cmax
ng/mL
Predicted Human 
Mean AUC
ng.h/mL
4 Week Oral Dog Repeat Dose Toxicity Study*
Most Likely Highest 
Potential
Most 
Likely
Highest 
Potential
Safety Margins for
Cmax
Safety Margins for 
AUC
2 mg
0.006
0.036
0.04
0.26
742 to > 9500X
155-1923X
10 mg
0.029
0.36
0.211
2.6
74 – 920X
16-192X
50 mg
0.15
1.8
1.09
13
15-178X
3.1-37X
200 mg
0.579
7.11
4.36
52
3.8-46X
0.8-9.3X
400 mg
1.16
14.2
8.72
104
1.9-23X
0.4-4.6X
*Safety margins are calculated with the dog NOAELs from the 1 month repeat dose toxicity study because mean Cmax 
was greater (26.7 ng/mL) than in rats (7.7 ng/mL) and the AUCs were similar (40.4 and 51 ng.h/mL for dogs and rats, 
respectively).
Although a negative food effect is predicted for this BCS III compound, the dose selected 
to evaluate the food effect will allow for a 2X increase in bioavailability due to the slight 
increase in bioavailability observed in the rat food effect study.
 
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6.
STUDY POPULATION
Prospective approval of protocol deviations to recruitment and enrollment criteria, also 
known as protocol waivers or exemptions, is not permitted.
6.1.
Inclusion Criteria
Participants are eligible to be included in the study only if all of the following criteria 
apply:
AGE
1
Between 18 and 55 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 the investigator based on a medical evaluation including 
medical history, physical examination, laboratory tests and cardiac monitoring.  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 Medical Monitor, agree and document that the finding is unlikely to 
introduce additional risk factors and will not interfere with the study procedures. 
3
History of regular bowel habits and no more than 3 bowel movements per day
SEX
4
Male or Female
a. Male participants:
A male participant must agree to use contraception as detailed in Section 12.7 of this 
protocol during the treatment period and for at least 7 days, corresponding to time 
needed to eliminate study treatment for both genotoxic and teratogenic study 
treatments after the last dose of study treatment and refrain from donating sperm 
during this period.
b. Female participants: 
A female participant is eligible to participate if she is not a woman of childbearing 
potential (WOCBP) as defined in Section 12.7.
INFORMED CONSENT
5
Capable of giving signed informed consent as described in Section 12.5 which
includes compliance with the requirements and restrictions listed in the informed 
consent form (ICF) and in this protocol.
 
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6.2.
Exclusion Criteria
Participants are excluded from the study if any of the following criteria apply:
CONCURRENT CONDITIONS/MEDICAL HISTORY (INCLUDES LIVER 
FUNCTION AND QTc INTERVAL) 
1.
ALT and bilirubin >1.5xULN (isolated bilirubin >1.5xULN is acceptable if bilirubin 
is fractionated and direct bilirubin <35%).
2.
Previous Diagnosis of IBS
3.
Estimated Glomerular Filtration Rate < 60 mL/min/1.73m2
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.
History of GERD, dyspepsia, GI bleeding, diverticulitis, diverticular stricture or 
other intestinal strictures, GI surgery that could affect motility, bezoars, dysphagia to 
solid food or pills, pelvic surgery within the past 3 months.
6.
Cardiac pacemakers or other implanted electromedical devices.
7.
Presence of hepatitis B surface antigen (HBsAg) at screening or within 3 months 
prior to first dose of study treatment
8.
Positive hepatitis C antibody test result at screening or within 3 months prior to first 
dose of study treatment.  NOTE: Participants with positive Hepatitis C antibody due 
to prior resolved disease can be enrolled, only if a confirmatory negative Hepatitis C 
RNA test is obtained
9.
A positive pre-study drug/alcohol screen.  
10. A positive test for HIV antibody.
11. History of regular alcohol consumption within 6 months of the study defined as: 
an average weekly intake of >14 standard drinks.  One standard drink is 
equivalent to 10 g of alcohol: 285 ml of beer, 100 ml of wine or 30 ml of 40% 
alcohol by volume distilled spirits. 
12. Current smoker or use of nicotine containing products within the past 3 months or 
unable to abstain from smoking tobacco or the use of nicotine-containing products 
while on study or positive pre-study cotinine test.
13. Unable to refrain from consumption of red wine, seville oranges, grapefruit or 
grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or fruit 
juices from 2 days prior to the first dose of Study Drug until inpatient check-out from 
the clinic.
14. QTcF>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.
 
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
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
15. 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 GSK 
Medical Monitor, contraindicates their participation. 
16. Unable to refrain from the use of prescription or non-prescription drugs, including 
vitamins, antacids, 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 for each dosing, 
unless in the opinion of the Investigator and GSK Medical Monitor the medication 
will not interfere with the study procedures or compromise subject safety.  
Paracetamol (≤ 2 grams per day) is acceptable.
RELEVANT HABITS
17. 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: 30 days, 5 half-lives or twice the duration of the biological effect of the 
investigational product (whichever is longer).
18. Exposure to more than 4 investigational medicinal products within 12 months prior 
to the first dosing day
19. Unwillingness or inability to follow the procedures outlined in the protocol.
20. Where participation in the study would result in donation of blood or blood products 
in excess of 500 mL within a 56 day period.
6.3.
Lifestyle Restrictions
6.3.1.
Part A Fasted: Meals and Dietary Restrictions

Subjects must fast from all food and drink (except water) for at least 8 hours 
prior to dose and PK sampling on Day 1 and will remain fasted for 
approximately 4 hours postdose.  Water may be consumed ad libitum.

Subjects will consume a standard weight neutral-meal for all meals and with no 
time limits.

Each dose of study drug will be administered with 240 mL of water while the 
subject is sitting/ standing.  All of the water must be completely consumed 
 
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within approximately 5 minutes.  Subjects may sit upright or stand, but must not 
lie down for at least 30 minutes after study drug administration.

Subjects must refrain from consumption of red wine, Seville oranges, grapefruit 
or grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or 
fruit juices from 2 days prior to the first dose of Study Drug on Day 1 until 
inpatient check-out from the clinic.
6.3.2.
Part A Fed for Pilot Food Effect (Cohort 2, Dosing Period 2):  
Meals and Dietary Restrictions

Subjects must fast from all food and drink (except water) for at least 8 hours 
prior to dose and PK sample on Day 1. Water may be consumed ad libitum.

Subjects will consume a mixed meal breakfast (also referred to as the PD meal
in Part B instructions) immediately following the dose on Day 1.  This identical 
mixed meal breakfast will consist of 55% carbohydrate, 15% protein, and 30% 
fat, representing 20% of subject’s total calorie intake.

The mixed meal breakfast must be eaten within 15 minutes and eaten in total.

Subjects will consume a standard weight-neutral meal for all other meals and 
with no time limits.

Each dose of study drug will be administered with 240 mL of water while the 
subject is sitting/ standing.  All of the water must be completely consumed 
within approximately 5 minutes.  Subjects may sit upright or stand, but must not 
lie down for at least 30 minutes after study drug administration.

Subjects must refrain from consumption of red wine, Seville oranges, grapefruit 
or grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or 
fruit juices from 2 days prior to the first dose of Study Drug on Day 1 until 
inpatient check-out from the clinic.
6.3.3.
Part B QD Fasted:  Meals and Dietary Restrictions

On PK days (Day 1, Day 7 and Day 14) Subjects must fast from all food and 
drink (except water) for at least 8 hours prior to dose and PK sample.  Subjects 
will remain fasted until mixed meal (PD) breakfast meal at approximately 2 
hours postdose.  Water may be consumed ad libitum.

On PD days (Day -1, Day 1 and Day 14) subjects must fast from all food and 
drink (except water) for at least 8 hours prior to PD sample.  Subjects will 
remain fasted until breakfast at approximately 2 hours postdose.  Water may be 
consumed ad libitum.

On the PD days (Day -1, Day 1, and Day 14) at approximately 2 hours postdose,
an identical mixed meal breakfast consisting of 55% carbohydrate, 15% protein, 
and 30% fat, representing 20% of subject’s total calorie intake will be 
consumed.

The PD breakfast meal must be eaten within 15 minutes and eaten in total.
 
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
The evening before PD testing days (Day -2, Day -1 and Day 13), subjects will 
consume a standard dinner, identical on all days.

Subjects will consume a standard weight neutral-meal for all other meals on PK 
and PD days as well as non PK and PD days with no time limits.

Lunch will be eaten approximately 4 hours after breakfast and dinner 
approximately 5 hours after lunch.

Each dose of study drug will be administered with 240 mL of water while the 
subject is sitting/ standing.  All of the water must be completely consumed 
within approximately 5 minutes.  Subjects may sit upright or stand, but must not 
lie down for at least 30 minutes after study drug administration.

Subjects must refrain from consumption of red wine, Seville oranges, grapefruit 
or grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or 
fruit juices from 2 days prior to the first dose of Study Drug on Day 1 until 
inpatient check-out from the clinic.
6.3.4.
Part B BID Fasted:  Meals and Dietary Restrictions

On PK days (Day 1, Day 7 and Day 14) Subjects must fast from all food and 
drink (except water) for at least 8 hours prior to AM dose and PK sample.  
Subjects will remain fasted until mixed meal (PD) breakfast meal at 
approximately 2 hours postdose.  Water may be consumed ad libitum.

On PK days (Day 1, Day 7 and Day 14) subjects must fast from all food and 
drink (except water) for at least 1 hour after PM dose administration.  Water 
may be consumed ad libitum.

On PD days (Day -1, Day 1 and Day 14) subjects must fast from all food and 
drink (except water) for at least 8 hours prior to PD sample.  Subjects will 
remain fasted until breakfast at approximately 2 hours postdose.  Water may be 
consumed ad libitum.

On PD days (Day -1, Day 1, and Day 14) subjects will receive an identical 
mixed meal breakfast at 1000 consisting of 55% carbohydrate, 15% protein, and 
30% fat, representing 20% of subject’s total calorie intake will be consumed.

The PD breakfast meal must be eaten within 15 minutes and eaten in total.

The evening before PD testing days (Day -2, Day -1 and Day 13), subjects will 
consume a standard dinner, identical on all days.

Lunch will be eaten at approximately 4 hours after breakfast meal and dinner 
approximately 5 hours after lunch.

Each dose of study drug will be administered with 240 mL of water while the 
subject is sitting/ standing.  All of the water must be completely consumed 
within approximately 5 minutes.  Subjects may sit upright or stand, but must not 
lie down for at least 30 minutes after study drug administration.

Subjects must refrain from consumption of red wine, Seville oranges, grapefruit 
or grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or 
 
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fruit juices from 2 days prior to the first dose of Study Drug on Day 1 until 
inpatient check-out from the clinic.
6.3.5.
Part B QD Fed:  Meals and Dietary Restrictions

On PK days (Day 1, Day 7 and Day 14) Subjects must fast from all food and 
drink (except water) for at least 8 hours prior to dose and PK sample.  Subjects 
will consume a PD breakfast meal immediately following dosing.  Water may be 
consumed ad libitum.

On PD days (Day -1, Day 1 and Day 14) subjects must fast from all food and 
drink (except water) for at least 8 hours prior to PD sample.  Subjects will 
consume breakfast immediately following dosing.  Water may be consumed ad 
libitum.

On the PD days (Day -1, Day 1, and Day 14), an identical mixed meal breakfast 
consisting of 55% carbohydrate, 15% protein, and 30% fat, representing 20% of 
subject’s total calorie intake will be eaten immediately following administration 
of study drug.

The PD breakfast meal must be eaten within 15 minutes and eaten in total.

The evening before PD testing days (Day -2, Day -1 and Day 13), subjects will 
consume a standard dinner, identical on all days.

Subjects will consume a standard weight-neutral meal for all other meals on PK 
and PD days as well as non PK and PD days and with no time limits.

Lunch will be eaten at approximately 4 hours after breakfast and dinner 
approximately 6 hours after lunch.

Each dose of study drug will be administered with 240 mL of water while the 
subject is sitting/ standing.  All of the water must be completely consumed 
within approximately 5 minutes.  Subjects may sit upright or stand, but must not 
lie down for at least 30 minutes after study drug administration.

Subjects must refrain from consumption of red wine, Seville oranges, grapefruit 
or grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or 
fruit juices from 2 days prior to the first dose of Study Drug on Day 1 until 
inpatient check-out from the clinic.
6.3.6.
Part  B BID Fed:  Meals and Dietary Restrictions

On PK days (Day 1, Day 7 and Day 14) Subjects must fast from all food and 
drink (except water) for at least 8 hours prior to dose and PK sample.  Subjects 
will consume the PD breakfast meal immediately following dosing.  Water may 
be consumed ad libitum.

On PD days (Day -1, Day 1 and Day 14) subjects must fast from all food and 
drink (except water) for at least 8 hours prior to PD sample.  Subjects will 
consume breakfast immediately following dosing.  Water may be consumed ad 
libitum.
 
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
On the PD days (Day -1, Day 1, and Day 14), an identical mixed meal breakfast 
consisting of 55% carbohydrate, 15% protein, and 30% fat, representing 20% of 
subject’s total calorie intake will be consumed.

The PD breakfast meal must be eaten within 15 minutes and eaten in total.

The evening before PD testing days (Day -2, Day -1 and Day 13), subjects will 
consume a standard dinner, identical on all days.

Subjects will consume a standard weight neutral meal for all other meals on PK 
and PD days as well as non PK and PD days and with no time limits.

Lunch will be eaten at approximately 4 hours after breakfast and dinner 
approximately 6 hours after lunch.

Each dose of study drug will be administered with 240 mL of water while the 
subject is sitting/ standing.  All of the water must be completely consumed 
within approximately 5 minutes.  Subjects may sit upright or stand, but must not 
lie down for at least 30 minutes after study drug administration.

Subjects must refrain from consumption of red wine, Seville oranges, grapefruit 
or grapefruit juice and/or pummelos, exotic citrus fruits, grapefruit hybrids or 
fruit juices from 2 days prior to the first dose of Study Drug on Day 1 until 
inpatient check-out from the clinic.
6.3.7.
Caffeine, Alcohol, and Tobacco

Subjects will abstain from alcohol for 24 hours prior to Day -1 to the clinic until 
inpatient check-out from the clinic.

The use of nicotine-containing products (including nicotine patches) will not be 
permitted from 3 months prior to admission to the clinic until inpatient check-out 
from the clinic.
6.3.8.
Activity

Participants will abstain from strenuous exercise for 48 hours before each blood
collection for clinical laboratory tests. Participants may participate in light 
recreational activities during studies (eg, watching television, reading).
6.4.
Screen Failures
Screen failures are defined as participants who consent to participate in the clinical study 
but are not subsequently randomized. A minimal set of screen failure information is 
required to ensure transparent reporting of screen failure participants to meet the 
Consolidated Standards of Reporting Trials (CONSORT) publishing requirements and to 
respond to queries from regulatory authorities. Minimal information includes 
demography, screen failure details, eligibility criteria, and any serious adverse events 
(SAEs).
Individuals who do not meet the criteria for participation in this study (screen failure) 
may be rescreened. 
 
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7.
TREATMENTS
Study treatment is defined as any investigational treatment or placebo intended to be 
administered to a study participant according to the study protocol.
7.1.
Treatments Administered
Study Treatment Name:
GSK3352589
Placebo
Dosage formulation:
Tablet
Tablet
Unit dose 
strength(s)/Dosage level(s):
1, 5, 25 and 100 mg
Placebos to match actives 
across all strengths
Route of Administration 
Oral
Oral
Dosing instructions:
Swallow whole with water, do 
not chew
Swallow whole with water, do 
not chew
Packaging and Labeling
Study Treatment will be labeled 
as required per country 
requirement. 
Study Treatment will be 
labeled as required per country 
requirement.
Manufacturer
WUXI/ GlaxoSmithKline
WUXI/ GlaxoSmithKline
7.2.
Dose Modification
This protocol allows some alteration from the currently outlined dosing schedule but the 
(predicted) maximum/cumulative exposure will not exceed a CMAX=26.6 ng/mL or 
AUC = 40.4 ng.h/mL.  The decision to proceed to the next dose level of GSK3352589 
(either an increase or a decrease) will be made by the Medical Monitor and the 
investigator based on safety, tolerability and preliminary PK data obtained in at least 4 
GSK3352589-treated subjects at the prior dose level.  The dosing schedule may also be 
adjusted to expand a dosing cohort to further evaluate safety or PK findings at a given 
dose level or to add cohorts to evaluate up to 2 additional dose levels or dosing regimens. 
The study procedures for these additional participants/cohorts will be the same as those 
described for other study participants/cohorts.   If moderate or severe AEs are 
consistently observed across participants in a cohort or if unacceptable pharmacological 
effects, reasonably attributable to study treatment in the opinion of the investigator are 
observed in more than 1 participant in a cohort, than dose escalation will be temporarily 
halted and no further participants will be dosed until a full safety review of the study has 
taken place.  Relevant reporting and discussion with Medical Monitor and the Ethics 
Committee will take place before resumption of dosing.  
 
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7.3.
Method of Treatment Assignment
The randomization schedule will be computer generated by GSK. On Day 1, subjects
will be assigned a unique number (randomization number) in ascending numerical order. 
The randomization number encodes the participant’s assignment to a treatment arm or 
sequence, according to the randomization schedule generated prior to the study by the 
Statistics Department at GSK. Each participant will be dispensed blinded study 
treatment, labeled with his/her unique randomization number, throughout the study.
7.4.
Blinding
This will be a double blind (sponsor unblind) study and the following will apply.  GSK 
staff will be considered unblinded to treatments, though the number with access to 
treatment information will be minimized. 

The investigator or treating physician may unblind a subject’s treatment assignment 
only in the case of an emergency OR in the event of a serious medical condition 
when knowledge of the study treatment is essential for the appropriate clinical 
management or welfare of the subject as judged by the investigator. 

Investigators have direct access to the subject’s individual study treatment. 

It is preferred (but not required) that the investigator first contacts the Medical 
Monitor or appropriate GSK study personnel to discuss options before unblinding the 
subject’s treatment assignment. 

If GSK personnel are not contacted before the unblinding, the investigator must 
notify GSK as soon as possible after unblinding, but without revealing the treatment 
assignment of the unblinded subject, unless that information is important for the 
safety of subjects currently in the study. 

The date and reason for the unblinding must be fully documented in the eCRF 
A subject will be withdrawn if the subject’s treatment code is unblinded by the 
investigator or treating physician. The primary reason for discontinuation (the event or 
condition which led to the unblinding) will be recorded in the eCRF.

GSK’s Global Clinical Safety and Pharmacovigilance (GCSP) staff may unblind 
the treatment assignment for any subject with an SAE. If the SAE requires that an 
expedited regulatory report be sent to one or more regulatory agencies, a copy of 
the report, identifying the subject’s treatment assignment, may be sent to 
investigators in accordance with local regulations and/or GSK policy.
7.5.
Preparation/Handling/Storage/Accountability
The investigator or designee must confirm appropriate temperature conditions have been 
maintained during transit for all study treatment received and any discrepancies are 
reported and resolved before use of the study treatment.
Only participants 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 
 
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in a secure, environmentally controlled, and monitored (manual or automated) area in 
accordance with the labeled 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 (ie, 
receipt, reconciliation, and final disposition records).
Further guidance and information for the final disposition of unused study treatment are 
provided in the Study Reference Manual.

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.
Precaution will be taken to avoid direct contact with the study treatment.  A Material 
Safety Data Sheet (MSDS) describing occupational hazards and recommended handling 
precautions will be provided to the investigator. In the case of unintentional occupational 
exposure notify the monitor, Medical Monitor and/or GSK study contact.
7.6.
Treatment Compliance
Participants are dosed at the site and 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 participant 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 participant’s mouth to ensure that the study 
treatment was ingested.
7.7.
Concomitant Therapy
Any medication or vaccine (including over-the-counter or prescription medicines, 
vitamins, and/or herbal supplements) that the participant is receiving at the time of 
enrolment or receives during the study must be recorded along with:

reason for use

dates of administration including start and end dates

dosage information including dose and frequency
The Medical Monitor should be contacted if there are any questions regarding 
concomitant or prior therapy.
Participants must abstain from taking prescription or nonprescription 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) before the start of study 
treatment until completion of the follow-up visit, unless, in the opinion of the investigator 
and GSK medical monitor, the medication will not interfere with the study. 
 
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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.
7.8.
Treatment after the End of the Study
Subjects will not receive any additional treatment from GSK after completion of the 
study because only healthy volunteers are eligible for study participation.
8.
DISCONTINUATION CRITERIA
8.1.
Discontinuation of Study Treatment
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. 
8.1.1.
Liver Chemistry Stopping Criteria
Liver chemistry stopping and increased monitoring criteria have been designed to 
assure participant safety and evaluate liver event etiology.  
Study treatment will be discontinued for a participant if liver chemistry stopping criteria 
are met:
 
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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
Section 12.8.
8.1.2.
QTc Stopping Criteria

The QTcF 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.

The QTc should be based on averaged QTcF values of triplicate 
electrocardiograms obtained over a brief (e.g., 5-10 minute) recording period.  
A subject that meets either bulleted criterion based on the average of triplicate ECG 
readings will be withdrawn from the study:

QTcF > 500 msec,

Change from baseline: QTcF >60 msec
See the SoA for data to be collected at the time of treatment discontinuation and follow-
up and for any further evaluations that need to be completed.
8.2.
Withdrawal from the Study

A participant may withdraw from the study at any time at his/her own request, 
or may be withdrawn at any time at the discretion of the investigator for safety, 
behavioural, compliance or administrative reasons.
 
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
If the participant withdraws consent for disclosure of future information, the 
sponsor may retain and continue to use any data collected before such a 
withdrawal of consent.

If a participant withdraws from the study, he/she may request destruction of any 
samples taken and not tested, and the investigator must document this in the site 
study records.

Refer to the SoA for data to be collected at the time of study discontinuation and
follow-up and for any further evaluations that need to be completed.
8.3.
Lost to Follow Up
A participant will be considered lost to follow-up if he or she repeatedly fails to return for 
scheduled visits and is unable to be contacted by the study site. 
The following actions must be taken if a participant fails to return to the clinic for a 
required study visit:

The site must attempt to contact the participant and reschedule the missed visit 
as soon as possible and counsel the participant on the importance of maintaining 
the assigned visit schedule and ascertain whether or not the participant wishes to 
and/or should continue in the study.

Before a participant is deemed lost to follow up, the investigator or designee 
must make every effort to regain contact with the participant (where possible, 3 
telephone calls and, if necessary, a certified letter to the participant’s last known 
mailing address or local equivalent methods). These contact attempts should be 
documented in the participant’s medical record.

Should the participant continue to be unreachable, he/she will be considered to 
have withdrawn from the study with a primary reason of lost to follow-up. 
9.
STUDY ASSESSMENTS AND PROCEDURES

Study procedures and their timing are summarized in the SoA. 

Protocol waivers or exemptions are not allowed 

Immediate safety concerns should be discussed with the sponsor immediately upon 
occurrence or awareness to determine if the participant should continue or 
discontinue study treatment.

Adherence to the study design requirements, including those specified in the SoA, is 
essential and required for study conduct.

All screening evaluations must be completed and reviewed to confirm that potential 
participants meet all eligibility criteria. The investigator will maintain a screening 
log to record details of all participants screened and to confirm eligibility or record 
reasons for screening failure, as applicable. 
 
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
Procedures conducted as part of the participant’s routine clinical management (eg, 
blood count) and obtained before signing of ICF may be utilized for screening or 
baseline purposes provided the procedure met the protocol-specified criteria and was 
performed within the time frame defined in the SoA.

The maximum amount of blood collected from each participant over the 
duration of the study, including any extra assessments that may be required, will 
not exceed 470 mL.

Repeat or unscheduled samples may be taken for safety reasons or for technical 
issues with the samples.
9.1.
Adverse Events
The definitions of an AE or SAE can be found in Section 12.6.
The investigator and any designees are responsible for detecting, documenting, and 
reporting events that meet the definition of an AE or SAE and remain responsible for 
following up AEs that are serious, considered related to the study treatment or the study,
or that caused the participant to discontinue the GSK3352589 (see Section 12.6). 
9.1.1.
Time Period and Frequency for Collecting AE and SAE 
Information

All SAEs will be collected from the signing of the ICF until the follow-up visit
at the time points specified in the SoA (Section 2).

All AEs will be collected from the start of treatment until the follow-up visit at 
the time points specified in the SoA (Section 2).

Medical occurrences that begin before the start of study treatment but after 
obtaining informed consent will be recorded on the Medical History/Current 
Medical Conditions section of the case report form (CRF) not the AE section.

All SAEs will be recorded and reported to the sponsor or designee within 24 
hours, as indicated in Section 12.6. The investigator will submit any updated 
SAE data to the sponsor within 24 hours of it being available.

Investigators are not obligated to actively seek AEs or SAEs in former study 
participants. However, if the investigator learns of any SAE, including a death, 
at any time after a participant has been discharged from the study, and he/she 
considers the event to be reasonably related to the study treatment or study 
participation, the investigator must promptly notify the sponsor.

The method of recording, evaluating, and assessing causality of AEs and SAEs
and the procedures for completing and transmitting SAE reports are provided in 
Section 12.6.
9.1.2.
Method of Detecting AEs and SAEs
Care will be taken not to introduce bias when detecting AE and/or SAE. Open-ended and 
non-leading verbal questioning of the participant is the preferred method to inquire about 
AE occurrence. 
 
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9.1.3.
Follow-up of AEs and SAEs
After the initial AE/SAE report, the investigator is required to proactively follow each 
participant at subsequent visits/contacts. All SAEs will be followed until the event is 
resolved, stabilized, otherwise explained, or the participant is lost to follow-up (as 
defined in Section 8.3). Further information on follow-up procedures is given in Section 
12.6.
9.1.4.
Regulatory Reporting Requirements for SAEs

Prompt notification by the investigator to the sponsor of a SAE is essential so 
that legal obligations and ethical responsibilities towards the safety of 
participants and the safety of a study treatment under clinical investigation are 
met. 

The sponsor has a legal responsibility to notify both the local regulatory 
authority and other regulatory agencies about the safety of a study treatment 
under clinical investigation. The sponsor will comply with country-specific 
regulatory requirements relating to safety reporting to the regulatory authority, 
Institutional Review Boards (IRB)/Independent Ethics Committees (IEC), and 
investigators.

Investigator safety reports must be prepared for suspected unexpected serious 
adverse reactions (SUSAR) according to local regulatory requirements and 
sponsor policy and forwarded to investigators as necessary.

An investigator who receives an investigator safety report describing a SAE or 
other specific safety information eg, summary or listing of SAE) from the 
sponsor will review and then file it along with the Investigator’s Brochure and
will notify the IRB/IEC, if appropriate according to local requirements.
9.1.5.
Pregnancy

Female subjects of child bearing potential are excluded from this study.

Details of all pregnancies in female participants and female partners of male 
participants will be collected for pregnancies occurring after the start of study 
treatment and until 7 days after the last dose.

If a pregnancy is reported, the investigator should inform GSK within 24 hours of
learning of the pregnancy and should follow the procedures outlined in Section 12.7.

Abnormal pregnancy outcomes (eg, spontaneous abortion, fetal death, stillbirth, 
congenital anomalies, ectopic pregnancy) are considered SAEs.
9.2.
Treatment of Overdose
For this study, any dose of GSK3352589 greater than 400 mg within a 24-hour time 
period 1 hour will be considered an overdose.
GSK does not recommend specific treatment for an overdose
 
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In the event of an overdose the investigator should:
1.
Contact the Medical Monitor immediately.
2.
Closely monitor the participant for AE/SAE and laboratory abnormalities until 
GSK3352589 can no longer be detected systemically (at least 7 days for 
GSK3352589).
3.
Obtain a plasma sample for PK analysis within 24 hours from the date of the last 
dose of study treatment 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 
participant.
9.3.
Safety Assessments
Planned time points for all safety assessments are provided in the SoA.  Additional time 
points for safety tests such as vital signs, physical exams and laboratory safety tests may 
be added during the course of the study based on newly available data to ensure 
appropriate safety monitoring.
9.3.1.
Physical Examinations

A complete physical examination will include, at a minimum, assessments of the 
Skin, Cardiovascular, Respiratory, Gastrointestinal and Neurological systems. 
Height and weight will also be measured and recorded.  Please refer to SoA as to 
when assessments are required.

A brief symptom directed physical examination will include, at a minimum, 
assessments of the skin, lungs, cardiovascular system, and abdomen (liver and 
spleen).

Investigators should pay special attention to clinical signs related to previous 
serious illnesses.
9.3.2.
Vital Signs

Vital signs will be measured in a semi-supine position after 5 minutes rest and 
will include temperature, systolic and diastolic blood pressure, and pulse rate.

Three readings of blood pressure and pulse will be taken. The first reading 
should be rejected. The second and third readings should be averaged to give the 
measurement to be recorded in the CRF
9.3.3.
Electrocardiograms

Triplicate 12-lead ECGs will be obtained at each timepoint during the study 
using an ECG machine that automatically calculates the heart rate and measures 
 
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PR, QRS, QT, and QTc intervals.  Refer to Section 8.1.2 for QTc withdrawal 
criteria and additional QTc readings that may be necessary

At each time point at which triplicate ECG are required, 3 individual ECG 
tracings should be obtained as closely as possible in succession, but no more 
than 2 minutes apart. The full set of triplicates should be completed in less than 
4 minutes.  The postdose timepoints for ECGs may be changed if PK data 
indicate an earlier or later tmax.
9.3.4.
Clinical Safety Laboratory Assessments

Refer to Section 12.3 for the list of clinical laboratory tests to be performed and 
to the SoA for the timing and frequency. 

The investigator must review the laboratory report, document this review, and 
record any clinically relevant changes occurring during the study in the AE 
section of the CRF. The laboratory reports must be filed with the source 
documents. Clinically significant abnormal laboratory findings are those which 
are not associated with the underlying disease, unless judged by the investigator 
to be more severe than expected for the participant's condition.

All laboratory tests with values considered clinically significantly abnormal 
during participation in the study or within 7 days after the last dose of study
treatment should be repeated until the values return to normal or baseline or are 
no longer considered significantly abnormal by the investigator or medical 
monitor. 

If such values do not return to normal/baseline within a period of time judged 
reasonable by the investigator, the etiology should be identified and the sponsor 
notified.

All protocol-required laboratory assessments, as defined in Section 12.3, must 
be conducted in accordance with the laboratory manual and the SoA.

If laboratory values from non-protocol specified laboratory assessments 
performed at the institution’s local laboratory require a change in participant 
management or are considered clinically significant by the investigator (eg, SAE 
or AE or dose modification), then the results must be recorded in the CRF.
9.3.5.
Bristol Stool Form Scale
The BSFS [Lewis, 1997] which describes 7 types of stool, will be used by the subject 
during the study.  Please refer to Section 12.4.
9.4.
Pharmacokinetics
9.4.1.
Sample Collection
9.4.1.1.
Blood sample collection
Blood samples for PK analysis of GSK3352589 will be collected at the time points 
indicated in Section 2, Schedule of Activities. The actual date and time of each blood 
 
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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. 
Collection details, Processing, storage and shipping procedures are provided in the SRM.
9.4.1.2.
Urine Sample Collection
Urine samples to aid in the structural identification of metabolites of GSK3352589 will
be collected at the timepoints listed in Schedule of Activities, Section 2. Details of urine 
sample processing, storage and shipping procedures are provided in the SRM.
9.4.2.
Sample Analysis
Plasma (PK) analysis will be performed under the control of PTS-IVIVT-BIB,
GlaxoSmithKline, the details of which will be included in the SRM. Concentrations of 
GSK3352589 will be determined in plasma samples using the currently approved 
bioanalytical methodology.  Raw data will be archived at the bioanalytical site (detailed 
in the SRM).
Plasma samples provided for metabolite analysis and any residual PK samples from the 
highest dose cohort in Part B (Days 1 and 14) will be analyzed for compound-related 
metabolites under a separate PTS-PD-Global Spectroscopy, GlaxoSmithKline protocol.  
In addition, urine samples will be collected pre-dose and over the post-dose time intervals 
0-24 hours (Days 1 and 14) and qualitatively analyzed for metabolites under a separate 
PTS-PD-Global Spectroscopy, GlaxoSmithKline protocol from the highest dose cohort 
only.
Genetic analyses will not be performed on any samples.
9.4.3.
Pharmacokinetic Analyses
Pharmacokinetic analysis will be the responsibility of the Clinical Pharmacology 
Sciences department within GlaxoSmithKline or designee. Plasma GSK3352589 
concentration-time data will be analyzed by non-compartmental methods with the most 
current version of Phoenix WinNonlin. Final calculations will be based on the actual 
sampling times recorded during the study. 
From the plasma concentration time data following single doses in Part A, the following 
pharmacokinetics parameters will be determined, as data permit, Cmax, Tmax, AUC(0-
last), AUC(0-24), AUC(0-infinity), t1/2 or MRT, Tlast, AUC extrapolated to infinity 
(AUCPEO), and Tlag for the food effect comparisons only.
From the plasma concentration time data following repeat doses in Part B, the following 
PK parameters will be determined, as data permit for both Days 1 and 14: maximum 
observed plasma concentration (Cmax) for both the dosing intervals and the 24 hour daily 
interval, Tmax for both dosing intervals and 24 hour daily interval, AUCs for both the 
dosing intervals (0-10h) or (0-24h) for both QD and BID repeat dose regimens. Tlast 
parameter will be included in the reporting. The Cmax and AUC parameters will be used 
 
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to assess dose proportionality in both Part A and B and accumulation in Part B as data 
permits.
9.5.
Pharmacodynamic Biomarkers
Subjects in Part B will undergo PD testing of total GLP-1 and total PYY levels after 
administration of a breakfast mixed-meal (PD meal) challenge on Day -1 (baseline),  Day 
1 and Day 14 at the end of the treatment period according to Section 2.  The timing of PD
samples may be altered and/or PD samples may be obtained on additional days or at 
additional time points. The samples for peptide analyses must be handled with extra care 
due to ex vivo degradation. Details on collection, processing, storage and shipping of 
blood samples are provided in the SRM.
9.6.
Genetics
Genetics are not evaluated in this study.
10.
STATISTICAL CONSIDERATIONS
10.1.
Sample Size Determination
A sufficient number of subjects will be screened to enroll 16 subjects for Part A and 48 
subjects (8 subjects/cohort) for Part B for a total of 64 randomized subjects. Sample sizes 
are based on feasibility.  No formal power calculations were performed, and no statistical 
sample size re-estimation or adjustment will be used.
10.2.
Populations for Analyses
For purposes of analysis, the following populations are defined:
Population
Description
Safety
All randomized participants who receive at least one dose of study 
medication. Participants will be analyzed according to the treatment 
they actually received.
PK Concentration
All subjects for whom a pharmacokinetic sample was obtained.
PK Parameter
All subjects in the PK Concentration Population who receive at least 
one active dose of GSK3352589 and provide pharmacokinetic 
parameters.
PD
All subjects who receive at least one dose of study drug with a 
baseline value and at least one postprandial PD biomarker value. 
 
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10.3.
Statistical Analyses
Safety and tolerability will be presented in tabular and/or graphical format and 
summarized descriptively.  No hypotheses will be tested.  Listings of adverse events and 
point estimates and 90% confidence intervals for numerical endpoints will be produced. 
BSFS will be summarized by frequency and proportion of stool types by subjects and by 
treatment arm.
Pharmacokinetic data will be presented in graphical and/or tabular form and will be
summarized descriptively. Descriptive statistics (n, arithmetic mean, standard deviation,
%CV, minimum, median, and maximum) will be calculated for all pharmacokinetic
parameters by treatment. For log-transformed variables, geometric mean, 95% confidence
interval, and %CVb (100 * √ (exp(SD2) -1)) will be provided, where the SD is the
standard deviation of log-transformed data. All pharmacokinetic data will be stored in the 
Archives, GlaxoSmithKline Pharmaceuticals, R&D. Data permitting, the PK parameter 
data will be presented descriptively (e.g. n, arithmetic mean, median, minimum, 
maximum) by treatment and part. Further summaries, analyses, and/or graphs may be 
produced as appropriate to the data.
Statistical analyses of the pharmacokinetic data will be the responsibility of Clinical 
Statistics, GlaxoSmithKline or designee.  In Part A, dose proportionality of GSK3352589 
will be analyzed by using appropriate power and ANOVA models.  Food effect analyses 
will be done according to the FDA guidance standards. In Part B, the Cmax and AUC 
parameters will be used to assess accumulation and dose proportionality. 
Analyses of PD data will be the responsibility of Clinical Statistics, GlaxoSmithKline or 
designee.  PD data for total GLP-1 and total PYY will be presented in graphical and/or 
tabular form and will be summarized descriptively.
If data permit, the relationship between GSK3352589 and postprandial levels will be 
explored.  PK/PD analyses will be performed to determine if relationship(s) between dose 
and/or GSK3352589 concentrations and postprandial concentrations of GLP-1 and PYY 
levels exist. Graphical investigations will be initially explored and appropriate linear and 
non-linear models may be evaluated.
Biotransformation of GSK3352589 in plasma and urine will also be evaluated for the 
identification of any compound-derived metabolite(s).
10.3.1.
Other Analyses
Pharmacokinetic, pharmacodynamic, and biomarker exploratory analyses will be 
described in the reporting and analysis plan.
10.3.2.
Interim Analyses
No formal statistical interim analysis is planned for this study. Safety, tolerability, and 
pharmacokinetic data will be reviewed prior to each dose-escalation and prior to all 
within subject escalations.
 
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11.
REFERENCES
Airaksinen, M.S., and Saarma, M. (2002). The GDNF family: signalling, biological 
functions and therapeutic value. Nature reviews Neuroscience 3, 383-394.
Azpiroz F, Bouin M, Camilleri M, Mayer EA, Poitras P, Serra J, Spiller RC. Mechanisms 
of Hypersensitivity in Irritable Bowel Syndrome and Functional Disorders. 
Neurogastroenterol Motil. 2007 Jan;19(1 Suppl):62-88.
Barrenschee, M., Bottner, M., Hellwig, I., Harde, J., Egberts, J.H., Becker, T., and 
Wedel, T. (2013). Site-specific gene expression and localization of growth factor ligand 
receptors RET, GFRalpha1 and GFRalpha2 in human adult colon. Cell and tissue 
research 354, 371-380.
Berman, S.M., Chang, L., Suyenobu, B., Derbyshire, S.W., Stains, J., Fitzgerald, L., 
Mandelkern, M., Hamm, L., Vogt, B., Naliboff, B.D., et al. (2002). Condition-specific 
deactivation of brain regions by 5-HT3 receptor antagonist Alosetron. Gastroenterology 
123, 969-977.
Camilleri, M. (2012). Peripheral mechanisms in irritable bowel syndrome. The New 
England journal of medicine 367, 1626-1635.
DeSesso, J.M. and Jacobson, C.F., Anatomical and physiological parameters affecting 
gastrointestinal absorption in humans and rats.  Food and chemical toxicology, 39:209-
228, 2001.
Dothel G, Barbaro MR, Boudin H, et. al. Nerve fiber outgrowth is increased in the 
intestinal mucosa of patients with irritable bowel syndrome. Gastroenterology. 2015 
May;148(5):1002-1011.e4
Edery P, Lyonnet S, Mulligan LM, Pelet A, Dow E, Abel L,. et al. Mutations of the RET
proto-oncogene in Hirschsprung’s disease. Nature. 1994;367:378-90.
GlaxoSmithKline Document Number 2016N305051_00: GSK3352589 Investigator’s 
Brochure. March 2017.
Guidance for industry, estimating the maximum safe starting dose in initial clinical trials 
for therapeutics in adult healthy volunteers, US Department of Health and Human 
Services, FDA, CDER, 2005. (http://www.fda.gov/ucm/groups/fdagov-public/@fdagov-
drugs-gen/documents/document/ucm078932.pdf)
Gyermek, L. (1996). Pharmacology of serotonin as related to anesthesia. Journal of 
clinical anesthesia 8, 402-425.
Halland, M., and Talley, N.J. (2013). New treatments for IBS. Nature reviews 
Gastroenterology & hepatology 10, 13-23.
Lewis SJ, Heaton KW, Stool form as a useful guide to intestinal transit time. Scand J 
Gastroenterol 1997; 32 (9) 920-924.
 
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Longstreth, G.F., Thompson, W.G., Chey, W.D., Houghton, L.A., Mearin, F., and Spiller, 
R.C. (2006). Functional bowel disorders. Gastroenterology 130, 1480-1491.
Physiological parameter values for PBPK models: a report by the international life 
sciences institute risk science institute.  U.S. Environmental Protection Agency, Office of 
Health and Environmental Assessment, 1994.
Plourde V, St-Pierre S, Quirion R. Calcitonin gene-related peptide in viscerosensitive 
response to colorectal distension in rats. Am. J. Physiol. 1997; 273:G191-6.
Radwan, A., Amidon, G.L., Langguth, P. Mechanistic investigation of food effect on 
disintegration and dissolution of BCS class III compound solid formulations: the 
importance of viscocity.  Biopharm. Drug Dispos. 33:403-416 (2012).
Tanaka T, Shinoda M, Feng B, et.al.  Modulation of visceral hypersensitivity by glial cell 
line-derived neurotrophic factor family receptor-3 in colorectal afferents. Am J Physiol 
Gastrointest Liver Physiol. 2011 Mar; 300(3):G418-24. doi: 10.1152/ajpgi.00456.2010. 
Epub 2010 Dec 30.
Wang H, Hughes I, Planer W, Parsadanian A, Grider JR, Vohra BPS et al.  The timing 
and location of glial cell line-derived neurotrophic factor expression determine enteric 
nervous system structure and function.  The Journal of Neuroscience, 2010;30:1523-38.
 
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12.
APPENDICES
12.1.
Appendix 1: Abbreviations and Trademarks
AE
Adverse Event
ALT
Alanine Transferase
ANOVA
Analysis of variance
AST
Aspartate Transferase
AUC
Area Under the Curve
AUCtau
Area under the plasma concentration-time curve over the 
dosing interval (0-4), (0-10), and  (0-24) hours 
AUClastor AUC (0-t)
Area under the plasma concentration-time curve from
time zero to last measurable time point
BCS
Biopharmaceutical classification system
BID
Twice a Day or twice daily
BSA
Body Surface Area
BSFS
Bristol Stool Form Scale
BUN
Blood Urea Nitrogen
CDER
Center for Drug Evaluation and Research
CL/F
Clearance
CLs
Systemic clearance
Cmax
Maximum Observed Plasma Concentration
CONSORT
Consolidated Standards of Reporting Trials
CNS
Central Nervous System
CTT
Colonic Transit Time
CRF
Case Report Form
CV
Cardiovascular
CYP
Cytochrome P450
DDI
Drug-Drug Interaction
e.g.
exempli gratia (for example)
ECG
Electrocardiogram
ELISA
Enzyme-linked immunosorbent assay
ENS
Enteric Nervous System
F
Bioavailability 
FDA
(US) Food and Drug Administration
FSH
Follicle Stimulating Hormone
FTiH
First Time in Human
g
gram
GCP
Good Clinical Practice
GCSP
Global Clinical Safety and Pharmacovigilance
GET
Gastric Emptying Time
GI
Gastrointestinal
GSK
GlaxoSmithKline
h
hour
HbsAg
Hepatitis B Surface Antigen
hCG
Human Chorionic Gonadotropin
 
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HED
Human Equivalent Dose
Hep B
Hepatitis B
Hep C
Hepatitis C
hERG
Human ether-a-go-go-related gene
HIV
Human Immunodeficiency Virus
HRT
Hormone Replacement Therapy
IB
Investigator’s Brochure
IBS
Irritable Bowel Syndrome
IBS-C
Irritable Bowel Syndrome Constipation
IBS-D
Irritable Bowel Syndrome Diarrhea
IBS-M
Irritable Bowel Syndrome Mixed
IC50
Half Maximal Inhibitory Concentration
ICH
International Conference on Harmonisation
IEC
Independent Ethics Committee
INR
International Normalized Ratio
IP
Investigational Product
IRB
Institutional Review Board
IV
Intravenous
Ka
Absorption rate constant
kg
Kilogram
Ki
Inhibitory Constant
MABEL
Minimum anticipated biological effect level
MCH
Mean Corpuscular Hemoglobin
MCV
Mean Corpuscular Volume
MedDRA
Medical Dictionary for Regulatory Activities
mg
milligram
mL
milliliter
MLA
Mouse Lymphoma Assay
MRTlast
Mean residence time up to last measurable concentration
MSDS
Material Safety Data Sheet
msec
Millisecond
ng/g
Nanogram per gram
ng/mg
Nanogram per milligram
nM
Nanometer
NOAEL
No Observed Adverse Effect Level
PAD
Pharmacological active dose
PD
Pharmacodynamic
PK
Pharmacokinetic
PI
Percentage Interval
PV
Pharmacovigiliance
QTc
Corrected QT Interval
QTcB
Corrected QT Interval to Bazett’s formula
QTcF
Corrected QT Interval to Fridericia’s formula 
RAP
Report and Analysis Plan
RBC
Red Blood Cell
RET
REarranged during Transfection
 
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RSA
Relative surface area
SAE
Serious Adverse Event
SBTT
Small Bowel Transit Time
SD
Standard Deviation
SGOT
Serum Glutamic Oxaloacetic Transaminase
SGPT
Serum Glutamic Pyruvic Transaminase
SOA
Schedule of Activities
SOP
Standard Operating Procedure
SRM 
Study Reference Manual
T1/2
Apparent Terminal Phase Half-Life
Tmax
Time to Maximum Observed Plasma Concentration
g
Microgram
ULN
Upper Limit of Normal
V/F
Volume of distribution
WBC
White Blood Cell
WGTT
Whole Gut Transit Time
WMC
Wireless Motility Capsule
Trademark Information
Trademarks of the GlaxoSmithKline 
group of companies
Trademarks not owned by the 
GlaxoSmithKline group of companies
ADVAIR
Chiron RIBA
Phoenix WinNonlin
SAS
 
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12.2.
Appendix 2: Human Pharmacodynamic Prediction Methods
A systems pharmacology model was developed for rats and then scaled to humans for
GSK3352589 in GI tract with a focus on colon lumen and colon tissue concentrations. 
Additional methods based on allometry concepts (body weight, BSA, and intestine 
surface areas) were applied for additional support and to facilitate expansion and account 
for error in the predicted dose range for this study. The dose range proposed for minimal 
pharmacological effect may be 5 mg once daily (2 mg twice daily) to 130 mg once or 
twice daily. The dose range for maximum pharmacological effect is predicted to be 5 mg 
to 440 mg.  It is unknown if these doses are best given once or twice daily, but twice 
daily dosing should provide more sustained tissue concentrations per day.  The rat IBS 
visceral hypersensitivity model indicates dosing to steady state tissue concentrations is 
required and that twice daily dosing is best in rats.  Efficacy with the rat PD colon 
hypersensitivity model correlates with a steady state colon tissue concentration of 
approximately 1218 ng/g (10 mg/kg for 3.5 days BID) and this target tissue concentration 
was assumed to be the same for human colon tissue.
12.2.1.
Systems pharmacology model of human colon tissue 
concentrations
Rat plasma profiles show little or no correlation with the rat model PD response. This 
presented a unique challenge as many established methods for determining dose/exposure 
response for a predetermined target concentration have been based on plasma drug 
concentrations. As colon tissue concentrations were determined to be the best predictor of 
patient response, it was determined a custom systems pharmacology modeling effort 
would be appropriate to address this challenge. A mechanistic model was developed to 
predict colon tissue concentrations as a function of colon content drug concentration. 
Development began with the determination of gut transit times, gut thickness, volumes 
and surfaces areas for rats and humans from the literature. Time course data from rat gut 
content and tissue concentrations were first used to estimate drug specific parameters, 
than once identified were fixed for subsequent human predictions. The model was used to 
simulate colon tissue concentrations for doses between 2 mg and 400 mg. Colon tissue 
time profile predictions were simulated to determine the time and duration above the 
target tissue concentration of 1218 ng/g following once daily (Figure 2) and twice daily 
(Figure 3). Following once daily oral doses of 7.5 mg and 10 mg and twice daily repeat 
dosing with 2 mg, 3 mg, and 5 mg, the colon tissue concentration remains above or near 
the target concentration of 1218 ng/g for the entire dosing interval. The duration of 
exposure needed or the time above 1218 ng/g is unknown.  
 
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Figure 2
Predicted human colon tissue concentration-time profiles following 
once daily dosing for four days ranging from 1 mg to 10 mg
Figure 3
Predicted human colon tissue concentration-time profiles following 
twice daily dosing for four days ranging from 1 mg to 5 mg
12.2.2.
Human Equivalent Dose (HED) Prediction Method using FDA 
guidance with and without small and large intestine surface area 
correction
This HED method is generally recommended for estimating safe starting doses in FTiH 
studies (FDA - CDER 2005 guidance), herein the conversion factors have been applied to 
the pharmacologically active doses (PAD). The HEDs are shown in Table 9. HED is 
 
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essentially a body surface area (BSA) correction. Because the rat and human have similar 
in vitro enzyme inhibition potency estimates, a potency adjustment was not needed.
Further correction was applied to the HED for small intestine surface area based on the
principle that more small intestine surface area will result in more absorption of drug in 
humans. It has been reported that the ratio of the human small intestine surface area 
normalized for BSA is 111. The same ratio for rat is 25. This comparison reveals that 
relative surface area of the human small intestine (RSA) is 4.4 times that of the rat 
[DeSesso, 2001]. The HED was divided by 4.4 and referred to as HED corrected for 
small intestine RSA and is considered a very conservative estimate.
In addition, correction was applied to the HED for colon surface area based on the 
principle there is less large intestine surface area in human compared to rat.  It has been 
reported that the ratio of the human large intestine surface area normalized for BSA is 
0.19.  The same ratio for rat is 0.85. This comparison reveals that the large intestine 
relative surface area of the human is 0.22 times that of the rat [DeSesso, 2001]. The HED 
was divided by 0.22 and referred to as HED corrected for large intestine RSA and is 
considered the more aggressive estimate.
Using these prediction methods, human doses anticipated to have minimum 
pharmacological effect are predicted to be between 7 mg and 136 mg for a 60 kg human 
following repeat dosing to steady state. The doses anticipated to achieve maximum 
pharmacological effect are predicted to be between 23 mg and 454 mg for a 60 kg 
human.
Table 8
Human  Dose Predictions
Rat
(0.25kg)
HED
(60 kg)
HED corrected for 
small intestine
RSA (rat to 
human)
HED corrected 
for large intestine
RSA (rat to 
human)
3 mg/kg*
Minimal effect; Rat 
visceral 
hypersensitivity 
model
0.48 mg/kg
  30mg
7 mg
136 mg
10mg/kg*
Maximal effect; Rat 
visceral 
hypersensitivity 
model
1.61 mg/kg
100 mg
23 mg
454 mg
*dosed twice daily for 3.5 days (7 doses) to steady state
 
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12.2.3.
Human Dose Prediction for of GSK3352589 Adjusted for Colon 
Content and Gut Weight
The minimally effective dose in the IBS-hypersensitivity rat model was 3 mg/kg and the 
maximally effective dose was 10 mg/kg.  The estimated bioavailability is 0.18% for rat 
and approximately 0.16% for human; therefore 99.82% and 99.84% of the dose remains 
in the GI tract from oral to elimination. 
The calculations below assume that the amount of GSK3353589 per gram of whole GI 
tract tissue at doses linked to efficacy rat model will be the same amounts and doses 
needed for humans.  Assuming the target colon content for human is between 115,185 
ng/g and 384,000 ng/g, the corresponding human doses are 122 mg and 406 mg (Table 
9).
Table 9
Method for human colon content and gut weight estimation
Parameter
Rat
(0.25kg)
Rat
(0.25kg)
Human
(60 kg)
Human
(60 kg)
Dosed to steady state
3mg/kg
10mg/kg
2.03mg/kg
6.77mg/kg
Dose 
0.75 mg or 
750 µg
2.5mg or 
2500 µg
122 mg
406 mg
99.82% of dose remaining in 
gut contents
748.7 µg
2496 µg
-
-
99.84% of dose remaining in 
gut contents
-
-
121,635 µg
405,504 µg
Whole GI tract (grams or mL) 
(stomach, small and large 
intestine)1
6.5 g
6.5 g
1056 g
1056 g
Calculated amount of drug in rat 
colon contents
115,185 
ng/g
384,000 
ng/g
-
-
Targeted amount of drug in 
human colon contents 
-
-
115,185 
ng/g
384,000 
ng/g
NOTES :
1.
Physiological parameter values for PBPK models: a report by the international life sciences institute risk science 
institute. U.S. Environmental Protection Agency, Office of Health and Environmental Assessment, 1994.
 
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12.3.
Appendix 3: Clinical Laboratory Tests

The tests detailed in Table 10 will be performed by the local laboratory. 

Protocol-specific requirements for inclusion or exclusion of participants are 
detailed in Section 6 of the protocol.

Additional tests may be performed at any time during the study as determined 
necessary by the investigator or required by local regulations.
Table 10
Protocol-Required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Hematology
Platelet Count
RBC Indices:
MCV
MCH
WBC count with 
Differential:
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils
RBC Count
Hemoglobin
Hematocrit
Clinical 
Chemistry1
BUN
Potassium
Aspartate 
Aminotransferase
(AST)/ Serum 
Glutamic-Oxaloacetic 
Transaminase
(SGOT)
Total and direct 
bilirubin
Creatinine
Sodium
Alanine 
Aminotransferase
(ALT)/ Serum 
Glutamic-Pyruvic 
Transaminase (SGPT)
Total Protein
Glucose -
nonfasting]
Calcium
Alkaline phosphatase
Albumin
Routine 
Urinalysis

Specific gravity

pH, glucose, protein, blood, ketones, [bilirubin, urobilinogen, nitrite, 
leukocyte esterase] by dipstick

Microscopic examination (if blood or protein is abnormal)
Other 
Screening 
Tests

HIV

Hepatitis B (HBsAg)

Hepatitis C (Hep C antibody)

FSH and estradiol (as needed in women of non-child bearing 
potential only)

Cotinine test

Alcohol and drug screen (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines)
 
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Laboratory 
Assessments
Parameters

Serum hCG Pregnancy test (Screening and follow up) and urine pregnancy 
test on Day -1.
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 8.1 and Section 12.8. All events of ALT 3 × upper limit of normal (ULN) 
and bilirubin 2 × ULN (>35% direct bilirubin) or ALT 3 × ULN and international normalized ratio (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).
2.
Local urine testing will be standard for the protocol unless serum testing is required by local regulation or IRB/IEC.
 
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12.4.
Appendix 4: Bristol Stool Form Scale
 
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12.5.
Appendix 5: Study Governance Considerations
Regulatory and Ethical Considerations

This study will be conducted in accordance with the protocol and with:

Consensus ethical principles derived from international guidelines 
including the Declaration of Helsinki and Council for International 
Organizations of Medical Sciences (CIOMS) International Ethical 
Guidelines

Applicable ICH Good Clinical Practice (GCP) Guidelines

Applicable laws and regulations

The protocol, protocol amendments, ICF, Investigator Brochure, and other 
relevant documents (eg, advertisements) must be submitted to an IRB/IEC by 
the investigator and reviewed and approved by the IRB/IEC before the study is 
initiated. 

Any amendments to the protocol will require IEC/IRB approval before 
implementation of changes made to the study design, except for changes 
necessary to eliminate an immediate hazard to study participants. 

The investigator will be responsible for the following:

Providing written summaries of the status of the study to the IRB/IEC 
annually or more frequently in accordance with the requirements, policies, 
and procedures established by the IRB/EC

Notifying the IRB/IEC of SAE or other significant safety findings as 
required by IRB/IEC procedures

Providing oversight of the conduct of the study at the site and adherence to 
requirements of 21 CFR, ICH guidelines, the IRB/IEC, European 
regulation 536/2014 for clinical studies (if applicable), and all other 
applicable local regulations
Informed Consent Process

The investigator or his/her representative will explain the nature of the study to 
the participant or his/her legally authorized representative and answer all 
questions regarding the study. 

Participants must be informed that their participation is voluntary. Participants 
or their legally authorized representative will be required to sign a statement of 
informed consent that meets the requirements of 21 CFR 50, local regulations, 
ICH guidelines, Health Insurance Portability and Accountability Act (HIPAA) 
requirements, where applicable, and the IRB/IEC or study center. 

The medical record must include a statement that written informed consent was 
obtained before the participant was enrolled in the study and the date the written 
consent was obtained. The authorized person obtaining the informed consent 
must also sign the ICF.
 
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
Participants must be re-consented to the most current version of the ICF(s) 
during their participation in the study. 

A copy of the ICF(s) must be provided to the participant or the participant’s 
legally authorized representative. 

Participants who are rescreened are required to sign a new ICF.
Data Protection

Participants will be assigned a unique identifier by the sponsor. Any participant 
records or datasets that are transferred to the sponsor will contain the identifier 
only; participant names or any information which would make the participant 
identifiable will not be transferred. 

The participant must be informed that his/her personal study-related data will be 
used by the sponsor in accordance with local data protection law. The level of 
disclosure must also be explained to the participant. 

The participant must be informed that his/her medical records may be examined 
by Clinical Quality Assurance auditors or other authorized personnel appointed 
by the sponsor, by appropriate IRB/IEC members, and by inspectors from 
regulatory authorities.
Publication Policy

The results of this study may be published or presented at scientific meetings. If 
this is foreseen, the investigator agrees to submit all manuscripts or abstracts to 
the sponsor before submission. This allows the sponsor to protect proprietary 
information and to provide comments.

The sponsor will comply with the requirements for publication of study results. 
In accordance with standard editorial and ethical practice, the sponsor will 
generally support publication of multicenter studies only in their entirety and not 
as individual site data. In this case, a coordinating investigator will be 
designated by mutual agreement.

Authorship will be determined by mutual agreement and in line with 
International Committee of Medical Journal Editors authorship requirements.
Dissemination of Clinical Study Data

Tabular study results will be posted on the US National Institutes of Health’s 
website ww.clintrials.gov and other publically accessible sites.
Data Quality Assurance

All participant data relating to the study will be recorded on printed or electronic 
CRF unless transmitted to the sponsor or designee electronically (eg, laboratory 
data). The investigator is responsible for verifying that data entries are accurate 
and correct by physically or electronically signing the CRF. 

The investigator must maintain accurate documentation (source data) that 
supports the information entered in the CRF. 
 
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
The investigator must permit study-related monitoring, audits, IRB/IEC review, 
and regulatory agency inspections and provide direct access to source data 
documents. 

The sponsor or designee is responsible for the data management of this study 
including quality checking of the data. 

Study monitors will perform ongoing source data verification to confirm that 
data entered into the CRF by authorized site personnel are accurate, complete, 
and verifiable from source documents; that the safety and rights of participants 
are being protected; and that the study is being conducted in accordance with the 
currently approved protocol and any other study agreements, ICH GCP, and all 
applicable regulatory requirements. 

Records and documents, including signed ICF, pertaining to the conduct of this 
study must be retained by the investigator for 25 years from the issue of the final 
Clinical Study Report (CSR)/ equivalent summary unless local regulations or 
institutional policies require a longer retention period. No records may be 
destroyed during the retention period without the written approval of the 
sponsor. No records may be transferred to another location or party without 
written notification to the sponsor. 
Source Documents

Source documents provide evidence for the existence of the participant and 
substantiate the integrity of the data collected. Source documents are filed at the 
investigator’s site.

Data reported on the CRF or entered in the eCRF that are transcribed from 
source documents must be consistent with the source documents or the 
discrepancies must be explained. The investigator may need to request previous 
medical records or transfer records, depending on the study. Also, current 
medical records must be available.

Definition of what constitutes source data can be found in the study reference 
manual (SRM).
Study and Site Closure
GSK or its designee reserves the right to close the study site or terminate the study at any 
time for any reason at the sole discretion of GSK. Study sites will be closed upon study 
completion. A study site is considered closed when all required documents and study 
supplies have been collected and a study-site closure visit has been performed.
The investigator may initiate study-site closure at any time, provided there is reasonable 
cause and sufficient notice is given in advance of the intended termination.
Reasons for the early closure of a study site by the sponsor or investigator may include 
but are not limited to:

Failure of the investigator to comply with the protocol, the requirements of the 
IRB/IEC or local health authorities, the sponsor's procedures, or GCP guidelines
 
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
Inadequate recruitment of participants by the investigator

Discontinuation of further study treatment development
 
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12.6.
Appendix 6: Adverse Events: Definitions and Procedures for 
Recording, Evaluating, Follow-up, and Reporting
Definition of AE
AE Definition

An AE is any untoward medical occurrence in a clinical study participant, temporally 
associated with the use of a study treatment, whether or not considered related to the 
study treatment.

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 study treatment.
Events Meeting the AE Definition 

Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (eg, ECG, radiological scans, vital signs measurements), 
including those that worsen from baseline, considered clinically significant in the 
medical and scientific judgment of the investigator (ie, not related to progression of 
underlying disease).

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 before the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected drug-drug 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 it is an intentional overdose taken with possible suicidal/self-
harming intent. Such overdoses should be reported regardless of sequelae.
Events NOT Meeting the AE Definition 

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 participant’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 
participant’s condition.

Medical or surgical procedure (eg, endoscopy, appendectomy): the condition that 
leads to the procedure is the AE.

Situations in which 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.
Definition of SAE
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (eg, hospitalization for signs/symptoms of the disease under study, 
death due to progression of disease).
A SAE is defined as any untoward medical occurrence that, at any dose:
a.
Results in death
b.
Is life-threatening
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
participant 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 inpatient hospitalization or prolongation of existing hospitalization
In general, hospitalization signifies that the participant 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 
outpatient setting. Complications that occur during hospitalization are AE. 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 persistent disability/incapacity

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 (eg, sprained ankle) which may interfere with or prevent 
everyday life functions but do not constitute a substantial disruption.
e.
Is a congenital anomaly/birth defect
f.
Other situations:

Medical or scientific judgment should be exercised in deciding whether SAE 
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 participant or may require medical or surgical intervention to prevent 
 
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one of the other outcomes listed in the above definition. These events should usually 
be considered serious.
Examples of such events include 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.
Recording AE and SAE
AE and SAE Recording

When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (eg, hospital progress notes, laboratory, and diagnostics reports) 
related to the event.

The investigator will then record all relevant AE/SAE information in the CRF.

It is not acceptable for the investigator to send photocopies of the participant’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 case, all participant identifiers, with the exception of the 
participant number, will be redacted on the copies of the medical records before 
submission to GSK.

The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information. Whenever possible, the diagnosis (not 
the individual signs/symptoms) will be documented as the AE/SAE.
Assessment of Intensity
The investigator will make an assessment of intensity for each AE and SAE reported 
during the study and assign it to 1 of the following categories: 

Mild: An event that is easily tolerated by the participant, causing minimal discomfort 
and not interfering with everyday activities.

Moderate: An event that causes sufficiently discomfort and interferes with normal 
everyday activities.

Severe: An event that prevents normal everyday activities. An AE that is assessed as 
severe should not be confused with an SAE. Severe is a category utilized for rating 
the intensity of an event; and both AE and SAE can be assessed as severe.
An event is defined as ‘serious’ when it meets at least 1 of the predefined outcomes 
as described in the definition of an SAE, NOT when it is rated as severe.
Assessment of Causality

The investigator is obligated to assess the relationship between study treatment and 
each occurrence of each AE/SAE as follows: unrelated, likely unrelated, possibly 
 
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related, probably related and related.

A "reasonable possibility" of a relationship conveys that there are facts, evidence, 
and/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 underlying disease(s), concomitant therapy, and other 
risk factors, as well as the temporal relationship of the event to study treatment 
administration will be considered and investigated.

The investigator will also consult the Investigator’s Brochure (IB) and/or Product 
Information, for marketed products, in 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 in which 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 before the initial transmission of the SAE data to GSK.

The investigator may change his/her opinion of causality in light of follow-up 
information and send an SAE follow-up report with the updated causality 
assessment.

The causality assessment is one of the criteria used when determining regulatory 
reporting requirements.
Follow-up of AE and SAE

The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as medically indicated or as requested by GSK to 
elucidate the nature and/or causality of the AE or SAE as fully as possible. This may 
include additional laboratory tests or investigations, histopathological examinations, 
or consultation with other health care professionals.

If a participant 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 24 hours of 
receipt of the information.
Reporting of SAE to GSK
SAE Reporting to GSK via Electronic Data Collection Tool

The primary mechanism for reporting SAE to GSK will be the electronic data 
 
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collection tool.

If the electronic system is unavailable for more than 24 hours, then the site will use 
the paper SAE data collection tool (see next section).

The site will enter the SAE data into the electronic system as soon as it becomes 
available.

The investigator or medically-qualified sub-investigator must show evidence within 
the eCRF (e.g., check review box, signature, etc.) of review and verification of the 
relationship of each SAE to IP/study participation (causality) within 72 hours of SAE 
entry into the eCRF.

After the study is completed at a given site, the electronic data collection tool will be 
taken off-line to prevent the entry of new data or changes to existing data.

If a site receives a report of a new SAE from a study participant or receives updated 
data on a previously reported SAE after the electronic data collection tool has been 
taken off-line, then the site can report this information on a paper SAE form (see 
next section) or to the medical monitor by telephone.

Contacts for SAE reporting can be found in the SRM and at the beginning of this 
protocol on the Sponsor/Medical Monitor Contact Information page.
 
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12.7.
Appendix 7: Contraceptive Guidance and Collection of 
Pregnancy Information
Definitions
Woman of Childbearing Potential (WOCBP)
Female participants
Female participants of childbearing potential are NOT eligible to participate. 
A woman is considered fertile following menarche and until becoming post-menopausal 
unless permanently sterile (see below)
Women in the following categories are not considered WOCBP
1. Premenarchal
2. Premenopausal female with ONE of the following:

Documented hysterectomy

Documented bilateral salpingectomy

Documented bilateral oophorectomy
Note:  Documentation can come from the site personnel’s: review of participant’s 
medical records, medical examination, or medical history interview.
3. Postmenopausal female

A postmenopausal state is defined as no menses for 12 months without an 
alternative medical cause. A high follicle stimulating hormone (FSH) level in 
the postmenopausal range may be used to confirm a postmenopausal state in 
women not using hormonal contraception or hormonal replacement therapy 
(HRT). However, in the absence of 12 months of amenorrhea, a single FSH 
measurement is insufficient. 

Females on HRT and whose menopausal status is in doubt will be required to 
use one of the non-hormonal highly effective contraception methods if they wish 
to continue their HRT during the study. Otherwise, they must discontinue HRT 
to allow confirmation of postmenopausal status before study enrollment.
Contraception Guidance
Male participants

Male participants with female partners of child-bearing potential are eligible to 
participate if they agree to ONE of the following during the protocol-defined 
time frame in Section 6.1 (up to 7 days after administration of the last dose of 
study drug): 
 
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
Are abstinent from penile-vaginal intercourse as their usual and preferred 
lifestyle (abstinent on a long term and persistent basis) and agree to remain 
abstinent 

Agree to use a male condom plus an additional method of contraception 
with a failure rate of <1% per year as described in Table 11 when having 
penile-vaginal intercourse with a woman of childbearing potential 

Men with a pregnant or breastfeeding partner must agree to remain abstinent 
from penile-vaginal intercourse or use a male condom during each episode of 
penile penetration for 7 days after study completion or from last dose

Refrain from donating sperm for duration of study and for 7 days after study 
completion or from last dose
For female partners of males participating in the study, highly effective contraceptive 
methods are described in the table below. 
Table 11
Highly Effective Contraceptive Methods
Highly Effective Contraceptive Methods That Are User Dependent a
Failure rate of <1% per year when used consistently and correctly. 
Combined (estrogen- and progestogen-containing ) hormonal contraception associated with 
inhibition of ovulationb

oral

intravaginal 

transdermal 
Progestogen-only hormonal contraception associated with inhibition of ovulationb

injectable 
Highly Effective Methods That Are User Independent 

Implantable progestogen-only hormonal contraception associated with inhibition of 
ovulationb

Intrauterine device (IUD)

Intrauterine hormone-releasing system (IUS)

bilateral tubal occlusion
Vasectomized partner 
(A vasectomized partner is a highly effective contraception method provided that the partner is 
the sole male sexual partner of the WOCBP and the absence of sperm has been confirmed. If 
not, an additional highly effective method of contraception should be used.)
Sexual abstinence 
(Sexual abstinence is considered a highly effective method only if defined as refraining from 
heterosexual intercourse during the entire period of risk associated with the study drug. The 
 
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reliability of sexual abstinence needs to be evaluated in relation to the duration of the study and 
the preferred and usual lifestyle of the participant.)
NOTES: 
a.
Typical use failure rates may differ from those when used consistently and correctly. Use should be consistent 
with local regulations regarding the use of contraceptive methods for participants in clinical studies. 
b.
Hormonal contraception may be susceptible to interaction with the study drug, which may reduce the efficacy of 
the contraceptive method. In this case two highly effective methods of contraception should be utilized during the 
treatment period and for at least 7 days  after the last dose of study treatment 
Collection of Pregnancy Information
Male participants with partners who become pregnant

Investigator will attempt to collect pregnancy information on any male participant’s 
female partner of a male study participant who becomes pregnant while participating 
in this study. This applies only to participants who receive study treatment.

After obtaining the necessary signed informed consent from the pregnant female 
partner directly, the investigator will record pregnancy information on the 
appropriate form and submit it to GSK within 24 hours of learning of the partner’s 
pregnancy. 

Partner will also be followed to determine the outcome of the pregnancy. 
Information on the status of the mother and child will be forwarded to GSK

Generally, follow-up will be no longer than 6 to 8 weeks following the estimated
delivery date. Any termination of the pregnancy will be reported regardless of fetal 
status (presence or absence of anomalies) or indication for procedure. 
Female Participants who become pregnant
Female participants who can become pregnant are not eligible to participate, 
however any female subject who becomes pregnant while participating will be 
withdrawn from the study.

Investigator will collect pregnancy information on any female participant, who 
becomes pregnant while participating in this study. 

Information will be recorded on the appropriate form and submitted to GSK within 
24 hours of learning of a participant's pregnancy. 

Participant will be followed to determine the outcome of the pregnancy. The 
investigator will collect follow up information on participant and neonate, which will 
be forwarded to GSK Generally, follow-up will not be required for longer than 6 to 8 
weeks beyond the estimated delivery date.  

Any termination of pregnancy will be reported, regardless of fetal status (presence or 
absence of anomalies) or indication for procedure. 

While pregnancy itself is not considered to be an AE or SAE, any pregnancy 
complication or elective termination of a pregnancy will be reported as an AE or 
SAE. 
 
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
A spontaneous abortion is always considered to be an SAE and will be reported as 
such.  

Any SAE occurring as a result of a post-study pregnancy which is considered 
reasonably related to the study treatment by the investigator, will be reported to GSK
as described in above. While the investigator is not obligated to actively seek this 
information in former study participants, he or she may learn of an SAE through 
spontaneous reporting. 
 
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12.8.
Appendix 8: 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 etiology. 
Phase I liver chemistry stopping criteria and required follow up assessments
Liver Chemistry Stopping Criteria 
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 
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

A specialist or hepatology consultation is 
recommended
If ALT≥3xULN AND bilirubin < 2xULN  and 
INR ≤1.5:

Viral hepatitis serology3

Obtain INR and recheck with each liver 
chemistry assessment until the 
transaminases values show downward 
trend

Obtain blood sample for pharmacokinetic 
(PK) analysis, obtained up to 5 days of last 
dose4 

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 

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 high 
performance liquid chromatography 
(HPLC) assay (quantifies potential 
acetaminophen contribution to liver injury 
in subjects with definite or likely 
acetaminophen use in the preceding 
week).

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 
PK 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. 
 
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