A Randomised, Double-blind, Placebo-controlled, Parallel Group, 8-week Treatment Study to Investigate the Safety, Pharmacodynamics, and Effect of the TLR7 Agonist, GSK2245035, on the Allergen-induced Asthmatic Response in Subjects With Mild Allergic Asthma
Phase: PHASE2
Conditions: Asthma
Sponsor: GlaxoSmithKline

2015N264314_04
CONFIDENTIAL
The GlaxoSmithKline group of companies
205540
1
TITLE PAGE
Division: Worldwide Development
Information Type: Protocol Amendment
Title:
A randomised, double-blind (sponsor open) placebo-controlled, 
parallel group, 8-week treatment study to investigate the safety, 
pharmacodynamics and effect of the TLR7 agonist, 
GSK2245035, on the allergen-induced asthmatic response in 
subjects with mild allergic asthma.
Compound Number:
GSK2245035
Development Phase:
II
Effective Date:
10-JUL-2017
Protocol Amendment Number:  04
Author (s):
 
Copyright 2017 the GlaxoSmithKline group of companies.  All rights reserved.  
Unauthorised copying or use of this information is prohibited.
PPD

2015N264314_04
CONFIDENTIAL
The GlaxoSmithKline group of companies
205540
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Revision Chronology
GlaxoSmithKline 
Document Number
Date
Version
2015N264314_00
2016-APR-06
Original
2015N264314_01
2016-JUN-15
Amendment No. 1
Section 4.6.1 and Section 7.1.1 have been adjusted slightly to make it clearer that subjects 
will be provided with a thermometer to be able to record temperature as part of capturing 
adverse event (AE) information.
In Section 5.2, exclusion criterion 13 has been updated to reflect the revised 
recommended alcohol consumption guidelines.
2015N264314_02
2016-AUG-17
Amendment No. 2
The protocol title and text have been amended to clarify that while the investigator and 
subjects are blinded the sponsor-is unblinded. 
Details of the procedures required at early withdrawal visits have been added to the Time 
and Events Table, Section 7.1.
Details around the Safety Review Team have been added to include a consultant external 
to GSK to Section 6.3.
The inclusion and exclusion criteria have been updated in Section 5.1 and Section 5.2. 
and clarification around prohibited medications has been provided in Section 6.11.2. to 
align with updated exclusion criteria (Section 5.2).
Administrative changes have been made to the T&E table (Section 7.1) and text to ensure 
consistency throughout.
2015N264314_03
2017-MAY-26
Amendment No. 3
Changes in the Primary Medical Monitor and contact details.
Alignment of contraception requirements in Section 5.1 and Section 12.5.1 for partners of 
male subjects that are female of reproductive potential. 
Section 5.1, Inclusion Criteria, has been updated to align with Section 12.5.1 
contraception requirements for partners of male subjects that are female of reproductive 
potential. Changes made to these sections.
Section 5.1 Inclusion Criteria, bullets have been replaced with numbers.

2015N264314_04
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2015N264314_04
2017-JUL-10
Amendment No. 4
Section 4.4 Design Justification, has been updated to permit earlier interim analysis.
Section 5.3, Screening Failures/Re-Screening, has been updated to allow re-screening.
Section 7.1.1 Time and Events Table – Screening Visit(s), has been updated to clarify 
timing of screening visits.
Section 10.2 Regulatory and Ethical Considerations, Including the Informed Consent 
Process, to clarify that a new ICF should be completed for each re-screened subject.

PPD
PPD

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MEDICAL MONITOR/SPONSOR INFORMATION PAGE
Medical Monitor/SAE Contact Information: 
Role
Name
Day Time Phone Number 
and email address
After-hours 
Phone/Cell/
Pager 
Number
Fax 
Number
Site Address
Primary 
Medical 
Monitor 
 
 
MD
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
Secondary 
Medical 
Monitor 
 
 MD
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
SAE contact  
information 
Medical 
monitor 
as above 
and 
Study 
Manager, 
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
Sponsor Legal Registered Address: 
GlaxoSmithKline Research & Development Limited
980 Great West Road
Brentford
Middlesex, TW8 9GS
UK
In some countries, the clinical trial sponsor may be the local GlaxoSmithKline Affiliate 
Company (or designee). If applicable, the details of the alternative Sponsor and contact 
person in the territory will be provided to the relevant regulatory authority as part of the 
clinical trial application.   
Regulatory Agency Identifying Number(s):  2015-005645-31
PPD
PPD
PPD
PPD
PPD
PPD

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INVESTIGATOR PROTOCOL AGREEMENT PAGE
For protocol 205540
I confirm agreement to conduct the study in compliance with the protocol, as amended by 
this protocol amendment.
I acknowledge that I am responsible for overall study conduct. I agree to personally 
conduct or supervise the described study.
I agree to ensure that all associates, colleagues and employees assisting in the conduct of 
the study are informed about their obligations. Mechanisms are in place to ensure that site 
staff receives the appropriate information throughout the study. 
Investigator Name:
Investigator Address:
Investigator Phone Number:
Investigator Signature
Date

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TABLE OF CONTENTS
PAGE
1.
PROTOCOL SYNOPSIS FOR STUDY 205540......................................................10
2.
INTRODUCTION....................................................................................................13
2.1.
Study Rationale/Brief background...............................................................13
3.
OBJECTIVE(S) AND ENDPOINT(S) ......................................................................14
4.
STUDY DESIGN ....................................................................................................16
4.1.
Overall Design ............................................................................................16
4.2.
Treatment Arms and Duration.....................................................................16
4.3.
Type and Number of Subjects.....................................................................16
4.4.
Design Justification.....................................................................................17
4.5.
Dose Justification........................................................................................18
4.6.
Benefit: Risk Assessment ...........................................................................19
4.6.1.
Risk Assessment .........................................................................20
4.6.2.
Benefit Assessment.....................................................................26
4.6.3.
Overall Benefit: Risk Conclusion..................................................26
5.
SELECTION OF STUDY POPULATION AND WITHDRAWAL CRITERIA .............26
5.1.
Inclusion Criteria.........................................................................................26
5.2.
Exclusion Criteria........................................................................................29
5.3.
Screening Failures/Re-screening................................................................31
5.4.
Withdrawal/Stopping Criteria.......................................................................31
5.4.1.
Liver Chemistry Stopping Criteria ................................................32
5.4.1.1.
Study Treatment Restart or Rechallenge....................32
5.4.2.
QTc Stopping Criteria ..................................................................32
5.5.
Subject and Study Completion....................................................................33
6.
STUDY TREATMENT ............................................................................................33
6.1.
Investigational Product and Other Study Treatment....................................33
6.2.
Treatment Assignment................................................................................34
6.3.
Blinding.......................................................................................................35
6.4.
Packaging and Labeling..............................................................................36
6.5.
Preparation/Handling/Storage/Accountability..............................................36
6.6.
Compliance with Study Treatment Administration.......................................37
6.7.
Treatment of Study Treatment Overdose....................................................37
6.8.
Treatment after the End of the Study ..........................................................37
6.9.
Challenge Agents .......................................................................................37
6.9.1.
Bronchial, intranasal and intra-dermal allergen 
challenge/administration ..............................................................37
6.10.
Lifestyle and/or Dietary Restrictions............................................................38
6.10.1.
Meals and Dietary Restrictions ....................................................38
6.10.2.
Caffeine, Alcohol, and Tobacco ...................................................38
6.10.3.
Activity .........................................................................................38
6.11.
Concomitant Medications and Non-Drug Therapies....................................38
6.11.1.
Permitted Medications and Non-Drug Therapies..........................38
6.11.2.
Prohibited Medications and Non-Drug Therapies.........................38
7.
STUDY ASSESSMENTS AND PROCEDURES .....................................................39
7.1.
Time and Events Tables .............................................................................41

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7.1.1.
Time and Events Table – Screening Visit(s) ................................41
7.1.2.
Time and Events Table – Dosing visit (DV) 1 (Day 1) –
DV8 (Week 8) and Early Withdrawal Visit (EW) ...........................43
7.1.3.
Time and Events Table – Follow up visits (FUV1 to 3) 
(Weeks 9 to 20) ...........................................................................45
7.2.
Screening and Critical Baseline Assessments ............................................47
7.2.1.
Skin prick testing..........................................................................47
7.3.
Efficacy.......................................................................................................47
7.3.1.
Bronchial allergen challenges and spirometry..............................47
7.3.2.
Intranasal and intra-dermal allergen challenge ............................47
7.3.3.
Sputum induction.........................................................................47
7.3.4.
TNSS...........................................................................................48
7.3.5.
Nasal examination .......................................................................48
7.3.6.
Fractional Exhaled Nitric Oxide (FeNO) .......................................48
7.4.
Safety .........................................................................................................49
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)..........49
7.4.1.1.
Time period and Frequency for collecting AE 
and SAE information...................................................49
7.4.1.2.
Method of Detecting AEs and SAEs ...........................50
7.4.1.3.
Follow-up of AEs and SAEs........................................50
7.4.1.4.
Cardiovascular and Death Events ..............................50
7.4.1.5.
Disease-Related Events and/or Disease-
Related Outcomes Not Qualifying as SAEs ................51
7.4.1.6.
Regulatory Reporting Requirements for SAEs............51
7.4.1.7.
Cytokine Related Symptoms (CRS) -
Symptoms Grading.....................................................52
7.4.2.
Pregnancy ...................................................................................52
7.4.3.
Physical Exams ...........................................................................52
7.4.4.
Vital Signs....................................................................................52
7.4.5.
Electrocardiogram (ECG).............................................................52
7.4.6.
Clinical Safety Laboratory Assessments ......................................53
7.4.7.
PEF monitoring............................................................................55
7.5.
Pharmacokinetics .......................................................................................55
7.5.1.
Blood Sample Collection..............................................................55
7.5.2.
Sample Analysis ..........................................................................55
7.6.
Exploratory Biomarkers...............................................................................55
7.6.1.
Pharmacodynamic Markers .........................................................55
7.6.1.1.
PD blood samples ......................................................55
7.6.1.2.
Blood PAX collection ..................................................56
7.6.1.3.
PD nasal lavage samples ...........................................56
7.6.1.4.
Bronchial allergen challenge (BAC) associated 
biomarkers in induced sputum samples......................56
7.6.1.5.
Bronchial allergen challenge (BAC) associated 
biomarkers in blood samples......................................56
7.6.1.6.
Nasal allergen challenge associated 
biomarkers in nasal fluid samples...............................56
7.6.1.7.
Cellular profile of nasal mucosa..................................56
7.6.2.
Novel Biomarkers ........................................................................57
7.7.
Genetics .....................................................................................................57
8.
DATA MANAGEMENT ...........................................................................................57

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9.
STATISTICAL CONSIDERATIONS AND DATA ANALYSES .................................58
9.1.
Hypotheses.................................................................................................58
9.2.
Sample Size Considerations.......................................................................58
9.2.1.
Sample Size Assumptions ...........................................................63
9.2.2.
Sample Size Sensitivity................................................................64
9.2.3.
Sample Size Re-estimation or Adjustment...................................64
9.3.
Data Analysis Considerations .....................................................................64
9.3.1.
Analysis Populations....................................................................64
9.3.2.
Interim Analysis ...........................................................................65
9.3.2.1.
Headline results for LAR endpoints at Follow 
Up 1 ...........................................................................67
9.4.
Key Elements of Analysis Plan ...................................................................68
9.4.1.
Primary Analyses.........................................................................68
9.4.2.
Secondary Analyses ....................................................................68
9.4.3.
Safety Analyses...........................................................................68
9.4.4.
Exploratory Analyses ...................................................................68
10. STUDY GOVERNANCE CONSIDERATIONS........................................................68
10.1.
Posting of Information on Publicly Available Clinical Trial Registers............68
10.2.
Regulatory and Ethical Considerations, Including the Informed 
Consent Process ........................................................................................69
10.3.
Quality Control (Study Monitoring) ..............................................................69
10.4.
Quality Assurance.......................................................................................70
10.5.
Study and Site Closure ...............................................................................70
10.6.
Records Retention......................................................................................71
10.7.
Provision of Study Results to Investigators, Posting of Information 
on Publically Available Clinical Trials Registers and Publication .................71
11. REFERENCES.......................................................................................................72
12. APPENDICES........................................................................................................73
12.1.
Appendix 1 – Abbreviations and Trademarks..............................................73
12.2.
Appendix 2: Liver Safety Required Actions and Follow up 
Assessments ..............................................................................................76
12.3.
Appendix 3 - Genetic Research ..................................................................78
12.4.
Appendix 4: Definition of and Procedures for Recording, Evaluating, 
Follow-Up and Reporting of Adverse Events...............................................81
12.4.1.
Definition of Adverse Events........................................................81
12.4.2.
Definition and Grading of CRS-related Adverse Events ...............83
12.4.3.
Definition of Serious Adverse Events...........................................84
12.4.4.
Definition of Cardiovascular Events .............................................86
12.4.5.
Recording of AEs and SAEs ........................................................86
12.4.6.
Evaluating AEs and SAEs............................................................87
12.4.7.
Reporting of SAEs to GSK...........................................................88
12.5.
Appendix 5: Modified List of Highly Effective Methods for Avoiding 
Pregnancy and Collection of Pregnancy Information...................................89
12.5.1.
Modified List of Highly Effective Methods for Avoiding 
Pregnancy ...................................................................................89
12.5.2.
Collection of Pregnancy Information ............................................89
12.5.3.
References ..................................................................................90
12.6.
Appendix 6 - Country Specific Requirements..............................................92
12.7.
Appendix 7: Protocol Amendment Changes................................................93

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1.
PROTOCOL SYNOPSIS FOR STUDY 205540
Rationale
GSK2245035 is a highly selective Toll-like receptor 7 (TLR7) agonist that preferentially 
stimulates the induction of type I interferons.  Intranasal (i.n.) administration of 
GSK2245035 in humans has been shown to cause immune changes in the upper airways 
milieu that may alter bystander immune responsiveness to aeroallergens and contribute to 
reduction of allergic reactivity in subjects with respiratory allergies.
In a 8-week repeat dose safety study (TL7116958) with i.n. GSK2245035 20 ng and 
80 ng, target engagement was confirmed by dose-related increases in serum and nasal 
lavage interferon inducible protein-10 (IP-10) levels in subjects with symptomatic 
allergic rhinitis and mild asthma. Prolonged activation (up to 1 week in the serum and 
3 weeks in nasal lavage) of the interferon (IFN) α pathway was noted at the higher dose 
with no associated adverse effects during this follow-up period.  Eight weekly doses of 
both 20 ng and 80 ng reduced nasal symptoms and allergic biomarkers after nasal 
allergen challenge and those reductions were sustained up to 3 weeks post treatment.
This study will assess whether TLR7-mediated pharmacology, with i.n. GSK2245035 
20 ng administered weekly for a period of 8 weeks, will lead to reduced allergic reactivity 
in the lower airways in subjects with mild allergic asthma.
Objective(s)/Endpoint(s)
Objectives
Endpoints
Primary

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on the allergen-induced late asthmatic 
response (LAR) in subjects with allergic 
asthma.

LAR: minimum forced expiratory volume 
in 1 second (FEV1) between 4-10 hours 
following allergen challenge one week 
after treatment.

LAR: weighted mean FEV1 between 4-10 
hours following allergen challenge one 
week after treatment.
Secondary

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on the allergen-induced early asthmatic 
response (EAR) in subjects with allergic 
asthma.

EAR: minimum FEV1 between 0-2 hours 
following allergen challenge one week 
after treatment.

EAR: weighted mean FEV1 between 0-2 
hours following allergen challenge one 
week after treatment.

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Objectives
Endpoints

Safety

To evaluate the safety and tolerability of 
treatment with eight weekly doses of 20 
ng i.n. GSK2245035

General safety endpoints including 
adverse events (AEs), peak expiratory 
flow (PEF), rescue salbutamol use, and 
clinical laboratory parameters

Exploratory

To evaluate the duration of effect of 
treatment with i.n. GSK2245035 
compared to placebo on the allergen-
induced asthmatic responses (LAR and 
EAR) in subjects with allergic asthma.

LAR: minimum and weighted mean 
FEV1 between 4-10 hours following 
allergen challenge approximately 4 and 
12 weeks after treatment.

EAR: minimum and weighted mean 
FEV1 between 0-2 hours following 
allergen challenge approximately 4 and 
12 weeks after treatment.

To evaluate the induction of TLR7-
associated pharmacodynamic (PD) 
biomarkers and exhaled nitric oxide 
(FeNO) following treatment with i.n. 
GSK2245035

TLR7-induced blood PD biomarkers.

TLR7 induced gene expression changes 
in blood

TLR7-induced nasal fluid PD biomarkers

Change in FeNO levels during the 
treatment phase

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Bronchial Allergen Challenge (BAC) 
associated allergic biomarkers

Allergic biomarkers in induced sputum

Allergic biomarkers in blood

Change in FeNO levels following BAC

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Nasal Allergen Challenge (NAC) 
associated allergic biomarkers

Allergic biomarkers in nasal fluids

Cellular profile of nasal mucosa

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Intradermal allergen challenge

Mean diameter early phase response

Mean diameter late phase response

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on total nasal symptoms

Total nasal symptoms score (TNSS) 
assessed daily (AM and PM) throughout 
the study treatment phase

To evaluate systemic pharmacokinetics 
(PK) after administration of i.n. 
GSK2245035 

Plasma GSK2245035 concentrations

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Overall Design
This will be a randomised, double-blind (sponsor open), placebo-controlled, parallel 
group, 8-week treatment study. Study schematic below:
Week -4
1       2      3     4     5      6      7      8       9
12 
20
Screening
8 week treatment period
Follow-up
BAC -Bronchial allergen challenge; NAC – Nasal allergen challenge/administration
IC – Intradermalchallenge; DV – Dosing Visit; FU – Follow-up
DV1  DV2 DV3 DV4 DV5 DV6 DV7 DV8   FU1          FU2             FU3
Randomisation
BAC/
NAC/IC
BAC/
NAC/
IC
BAC/
NAC/
IC
BAC
Treatment Arms and Duration

The study will consist of a screening period of up to approximately 4 weeks 
(involving two screening visits), a blinded treatment period of 8 weeks, followed by a 
follow-up period of up to 3 months.  The total duration of the study for each subject 
will therefore be a maximum of approximately 6 months. All doses will be 
administered in the clinic.

Approximately 24 subjects per arm will be randomised in a 1:1 ratio to receive either 
i.n. 20 ng GSK2245035 or placebo every week for the duration of 8 weeks.
Type and Number of Subjects

Subjects with allergic asthma who are sensitized to perennial or seasonal allergen(s) 
will be enrolled in the study.  Based on the restriction of concomitant medications as 
shown in the exclusion criteria, the target population includes subjects with mild 
asthma not requiring maintenance treatment with inhaled corticosteroid who are able 
to be treated with short acting beta2-agonist (SABA) alone for the duration of the 
study (up until the final follow-up visit).

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
A sufficient number of subjects will be enrolled to ensure that 20 per treatment arm 
complete the 8 week study treatment period and the BAC at the one week follow-up 
visit (one week after the 8th dose of study treatment).
Analysis
Statistical models will be fitted separately to each primary endpoint.  These endpoints 
will be analysed using analysis of covariance (ANCOVA) with treatment and baseline in 
the model; other covariates such as centre, allergen exposure (‘presumed’/’unknown’) 
and allergen exposure by treatment interaction may be included in the model.
Treatment comparisons of GSK2245035 vs Placebo will be presented with results also 
expressed as percentage attenuations to aid clinical interpretation. The posterior 
distribution for GSK2245035 vs Placebo will be obtained and expressed as percentage 
attenuation. The posterior probability of any percentage attenuation will be obtained, 
along with appropriate summary statistics from the posterior distribution (e.g. 70% equi-
tail Credible Intervals).
The principle inferences are expected to come from models with non-informative priors. 
Sensitivity analyses using informative priors based on AstraZeneca (AZ) data may be 
fitted [Leaker , 2012].
An interim analysis will be conducted to assess how many subjects on placebo did not 
experience an LAR as such subjects will reduce the chances of establishing the primary 
objective of this study.  This interim analysis will determine whether i) further 
recruitment into the trial should continue (i.e. a futility analysis), or ii) additional subjects 
should be enrolled to counteract the impact.
2.
INTRODUCTION
2.1.
Study Rationale/Brief background
The immunological milieu of the airways plays a critical role in shaping the nature of 
host responsiveness to inhaled allergens with predominant induction of aberrant Th2 
responses driving the development of respiratory allergies and asthma. The scientific 
hypothesis is that intranasal delivery of a TLR7 agonist will activate local dendritic cells, 
inducing a type 1 IFN response, and result in the modification of the nature of both B and 
T cell responses to allergen. Driving the development of Th1/Treg responses may reduce 
the magnitude of mucosal Th2 reactivity and reduce local IgE responses.
GSK2245035 is a highly selective Toll-like receptor 7 (TLR7) agonist that stimulates 
preferentially the induction of type I interferons.  In a 8-week repeat dose safety study 
(TL7116958) [GlaxoSmithKline Document Number 2013N174200_00] with i.n. 
GSK2245035 20ng and 80 ng, target engagement was confirmed by dose-related 
increases in serum and nasal lavage IP-10, (interferon inducible protein 10) levels in 
subjects with symptomatic allergic rhinitis and mild asthma.  Prolonged activation (up to 
1 week in the serum and 3 weeks in nasal lavage) of the IFNα pathway was noted at the 
higher dose with no associated adverse effects during this follow-up period.  Eight 
weekly doses of both 20 ng and 80 ng reduced nasal symptoms and allergic biomarkers 

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after nasal allergen challenge and those reductions were sustained up to 3 weeks post 
treatment.
Having demonstrated the effects i.n. GSK2245035 in patients with allergic rhinitis, this 
study will assess whether TLR7-mediated pharmacology, with i.n. GSK2245035 20 ng 
administered weekly for a period of 8 weeks, will lead to reduced allergic reactivity in the 
lower airways in subjects with mild allergic asthma using a bronchial allergen challenge 
model (BAC) at various time-points following treatment.  For asthma patients who have 
allergic triggers for their disease, an i.n. treatment may be considered ideal to maximize 
co-localization with both its target, local dendritic cells, and aeroallergen.  Clinical 
evidence to support the hypothesis that an intranasal TLR7 agonist can reduce the 
response to allergen in the lower airways using a BAC model has been reported (Leaker , 
2012). 
3.
OBJECTIVE(S) AND ENDPOINT(S)
Objective(s)/Endpoint(s)
Objectives
Endpoints
Primary

To evaluate the effect of treatment with i.n. 
GSK2245035 compared to placebo on the 
allergen-induced late asthmatic response 
(LAR) in subjects with allergic asthma.

LAR: minimum FEV1 between 4-10 hours 
following allergen challenge one week after 
treatment.

LAR: weighted mean FEV1 between 4-10 
hours following allergen challenge one 
week after treatment.
Secondary

To evaluate the effect of treatment with i.n.
GSK2245035 compared to placebo on the 
allergen-induced early asthmatic response 
(EAR) in subjects with allergic asthma.

EAR: minimum FEV1 between 0-2 hours 
following allergen challenge one week after 
treatment.

EAR: weighted mean FEV1 between 0-2 
hours following allergen challenge one 
week after treatment.

Safety

To evaluate the safety and tolerability of 
treatment with eight weekly doses of 20 ng 
i.n. GSK2245035

General safety endpoints including AE, 
peak expiratory flow (PEF), rescue 
salbutamol use, and clinical laboratory 
parameters

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Objectives
Endpoints

Exploratory

To evaluate the duration of effect of 
treatment with i.n. GSK2245035 compared 
to placebo on the allergen-induced 
asthmatic responses (LAR and EAR) in 
subjects with allergic asthma.

LAR: minimum and weighted mean FEV1
between 4-10 hours following allergen 
challenge approximately 4 and 12 weeks 
after treatment.

EAR: minimum and weighted mean FEV1
between 0-2 hours following allergen 
challenge approximately 4 and 12 weeks 
after treatment.

To evaluate the induction of TLR7-
associated PD biomarkers and FeNO 
following treatment with i.n. GSK2245035

TLR7-induced blood PD biomarkers.

TLR7 induced gene expression changes in 
blood

TLR7-induced nasal fluid PD biomarkers

Change in FeNO levels during the 
treatment period

To evaluate the effect of treatment with i.n. 
GSK2245035 compared to placebo on 
Bronchial Allergen Challenge (BAC) 
associated allergic biomarkers

Allergic biomarkers in induced sputum

Allergic biomarkers in blood

Change in FeNO levels following BAC

To evaluate the effect of treatment with i.n. 
GSK2245035 compared to placebo on 
Nasal Allergen Challenge (NAC) 
associated allergic biomarkers

Allergic biomarkers in nasal fluids

Cellular profile of nasal mucosa

To evaluate the effect of treatment with i.n. 
GSK2245035 compared to placebo on 
Intradermal allergen challenge

Mean diameter early phase response

Mean diameter late phase response

To evaluate the effect of treatment with i.n. 
GSK2245035 compared to placebo on total 
nasal symptoms

Total nasal symptoms score (TNSS) 
assessed daily (AM and PM) throughout 
the study treatment phase

To evaluate systemic pharmacokinetics 
(PK) after administration of i.n. 
GSK2245035 

Plasma GSK2245035 concentrations

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4.
STUDY DESIGN
4.1.
Overall Design

This will be a randomised, double-blind (sponsor open), placebo-controlled, parallel 
group, 8-week treatment study. Study schematic below:
Week -4
1       2      3     4     5      6      7      8       9
12 
20
Screening
8 week treatment period
Follow-up
BAC -Bronchial allergen challenge; NAC – Nasal allergen challenge/administration
IC – Intradermalchallenge; DV – Dosing Visit; FU – Follow-up
DV1  DV2 DV3 DV4 DV5 DV6 DV7 DV8   FU1          FU2             FU3
Randomisation
BAC/
NAC/IC
BAC/
NAC/
IC
BAC/
NAC/
IC
BAC
4.2.
Treatment Arms and Duration

The study will consist of a screening period of up to approximately 4 weeks 
(involving two screening visits), a blinded treatment period of 8 weeks, followed by a 
follow-up period of up to 3 months.  The total duration of the study for each subject 
will therefore be a maximum of approximately 6 months. All doses will be 
administered in the clinic.

Approximately 24 subjects per treatment arm will be randomised in a 1:1 ratio to 
receive either i.n. 20ng GSK2245035 or placebo every week for the duration of 8 
weeks.
4.3.
Type and Number of Subjects

Subjects with allergic asthma who are sensitized to perennial or seasonal allergen(s) 
will be enrolled in the study.  Based on the restriction of concomitant medications as 
shown in the exclusion criteria, the target population includes subjects with mild 
asthma not requiring maintenance treatment with inhaled corticosteroid who are able 

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to be treated with short acting beta2-agonist (SABA) alone for the duration of the 
study (up until the final follow-up visit).

A sufficient number of subjects will be enrolled to ensure that 20 per treatment arm 
complete the 8 week study treatment period and the BAC at the one week follow-up 
visit (one week after the 8th dose of study treatment).
If subjects prematurely discontinue the study, additional replacement subjects may be 
recruited and assigned the next available randomisation number, at the discretion of the 
Sponsor, in consultation with the investigator.
Following the interim analysis, additional subjects may also be enrolled to counteract the 
impact of individuals not exhibiting an LAR response on placebo.
Accounting for replacements and interim analysis recommendations, the total number of 
subjects to be randomised in this study is not expected to exceed 60.
4.4.
Design Justification

GSK2245035 is currently being developed for patients with allergic asthma.  It has 
been studied in healthy volunteers (HVT) and subjects with allergic rhinitis with and 
without mild asthma.  

This study will assess the safety, target engagement, and efficacy of GSK2245035 in 
subjects with allergic asthma using a placebo-controlled, parallel group design and a 
BAC model.  The BAC model is an established methodology and will be used in this 
study to determine whether 8 weekly doses of 20 ng i.n. GSK2245035 attenuates 
allergen-induced asthmatic responses compared to placebo.  In addition to the 
assessment after BAC at the end of the 8-week treatment (1 week after the 8th dose), 
additional BACs will be performed at approximately 4 weeks and 12 weeks after the 
8th dose to assess the durability of the effect.

In an 8-week repeat dose safety study TL7116958 [GlaxoSmithKline Document 
Number 2013N174200_00], there was a reduction of TNSS at15 minutes post-NAC. 
This could indicate an effect of GSK2245035 treatment on mast cell activation and 
mediator release. In the current study, nasal lining fluid will be collected before and 
up to 5 minutes after nasal allergen challenges to evaluate the impact of GSK2245035 
on mast cell mediator release. NACs will be performed approximately 24 hours after 
the BACs that are conducted at screening and at approximately 1 week and 4 weeks 
after the final dose.

Interim analysis: Previous experience within GSK suggests some individuals 
(approximately 20%) fail to demonstrate a LAR response after the screening visit (i.e. 
they pass the screening criteria but do not demonstrate another LAR response when 
receiving placebo during the on-treatment phase). Those GSK studies were mainly 
crossover designs with a placebo in every sequence, so subjects who did not 
demonstrate a post screening LAR could be retrospectively identified and excluded 
from the statistical analyses. However this study has parallel groups and the profiles 
of a “perfect” drug effect and an inconsistent LAR subject are indistinguishable so 
retrospective exclusions cannot be made for the GSK2245035 arm. If the rate of 
“non-reproducible LAR” subjects is large it will reduce the probability of success. 

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The purpose of the interim analysis is to evaluate whether the non-reproducible LAR 
rate is reducing the expected probability of success by more than 10% and, if it is, to 
perform an unblinded review of the observed data to assess the impact and cease 
further recruitment into the study, or enrol additional subjects to counteract the 
effects. Simulations were used to quantify the expected reduction in probability of 
success, using an estimate of the non-responder rate from the placebo arm only. The 
timing of the interim is linked to the enrolment rates and will be triggered by 
whichever occurs first i) Approximately 1 year after the first subject successfully 
completes the Follow Up 1 assessments or, ii) When 28 subjects have successfully 
completed the Follow Up 1 assessments or iii) At the discretion of the GSK Study 
Team up to the time that 28 subjects have successfully completed the Follow Up 1 
assessment.

It is hypothesized that TLR7-mediated immunomodulation requires allergen to be 
present and exposure to aeroallergens may fluctuate or remain relatively constant 
depending on the allergen, time of year, and a subject's household characteristics or 
activities.  Further, exposure to certain aeroallergens may be presumed and, in other 
instances, is unknown unless outdoor air and household samples are analyzed.  For 
example, allergen exposure may be presumed if a subject who is sensitized to and 
challenged with grass pollen enrolls during grass pollen season or whose treatment is 
expected to coincide with grass pollen season.  A subject sensitized to animal 
epithelia who has that animal in his/her house or has occupational exposure may be 
presumed to have exposure.  These subjects will potentially exhibit a different 
response to study drug to someone without similar exposure to the allergen.  
Exposure to certain allergens, particularly perennial allergens (i.e. house dust mite) 
cannot be presumed and will be considered unknown for the purposes of this 
protocol.  Therefore, the allergen a subject is administered for the BAC will need to 
be considered according to whether the subject has exposure to that allergen outside 
of the challenge.  Consequently, the randomization will be stratified by ‘presumed’ 
and ‘unknown’ exposure to allergen in addition to stratification by centre.
4.5.
Dose Justification
The 20ng weekly i.n. dose of GSK2245035 was selected based on previous single and 
repeat dose studies in healthy volunteers (HVTs) and allergic rhinitis subjects with and 
without mild asthma.

In the first time in human study TL7114450 (dose range 2 ng – 100 ng) 
[GlaxoSmithKline Document Number 2011N120740_00], single i.n. administration 
of doses up to 40 ng GSK2245035 was well tolerated in HVT and subjects with 
symptomatic seasonal allergic rhinitis.

In the 4-week repeat dose study TL7116392 [GlaxoSmithKline Document Number 
2011N126275_00], the repeat dose of i.n. GSK2245035 40 and 80 ng had an 
acceptable safety profile.  Adverse events (AEs) likely related to TLR7-induced 
cytokines, such as headache, fever, chills/rigors and nausea were observed in one 
subject receiving placebo, 4 of 6 subjects receiving 40 ng GSK2245035 and one of 7 
subjects receiving 80 ng GSK2245035.

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
In a 8-week repeat dose safety study TL7116958 [GlaxoSmithKline Document 
Number 2013N174200_00] with i.n. GSK2245035 in subjects with symptomatic 
allergic rhinitis and mild asthma, 20 ng and 80 ng weekly doses produced sustained 
reductions in nasal allergic reactivity based on the total nasal symptom score after 
nasal allergen challenge up to 3 weeks post treatment.  The tolerability of 20 ng 
GSK2245035 was similar to placebo with no reports of cytokine-related fever.  
Cytokine release syndrome (CRS)-related symptoms were self-resolving and occurred 
infrequently over the 8-week study period in subjects receiving 20 ng GSK2245035.  
GSK2245035 80 ng was poorly tolerated.  All but one subject receiving 80 ng 
experienced flu-like symptoms.  Although CRS-related symptoms were also self-
resolving at the 80 ng dose, subjects experienced them fairly consistently over the 
study period.  TLR7-induced biomarkers indicated target engagement at both doses, 
with evidence for sustained activation of TLR7-mediated pathway intranasally up to 3 
weeks post treatment only at the 80ng dose.  Reviewing the totality of data obtained 
in these previous studies supports further study of a repeated weekly dose of 20 ng 
i.n. GSK2245035.
4.6.
Benefit: Risk Assessment
Summaries of findings from both clinical and non-clinical studies conducted with 
GSK2245035 can be found in the Investigator’s Brochure [GlaxoSmithKline Document 
Number 2010N110564_04]. The following section outlines the risk assessment and 
mitigation strategy for this protocol:  

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4.6.1.
Risk Assessment
Potential Risk of Clinical Significance
Summary of Data/Rationale for Risk
Mitigation Strategy
Investigational Product (IP) [GSK2245035]
Events potentially related to cytokine 
induction.
The principal systemic effect seen with 
GSK2245035 in intranasal studies in 
animals (mice, rats, rabbits, and monkeys) 
up to 8 weeks in duration with weekly 
dosing was increases in the levels of 
specific cytokines; all other findings were 
considered to be secondary to this primary 
response (i.e., nasal irritancy, increased 
body temperature, increased heart rate, 
alterations in splenic and lymph node 
cellularity and minor changes in some 
clinical chemistry parameters).  With the 
exception of the splenic and lymph node 
effects, all the observed changes would be 
readily monitorable in humans.  The effects 
that were seen in the spleen and lymph node 
were of low severity and were shown to be 
fully reversible on withdrawal from 
treatment in the monkey.  The observed 
cytokine response is consistent with the 
known TLR7 pharmacology of 
GSK2245035.  Although a no effect level 
for cytokine response was not determined in 
the monkey, a No Observed Adverse Event 
Level (NOAEL) of 35.9 ng/kg/day was 
determined for the only secondary finding 
The study will be undertaken at experienced 
research units. The sites have all necessary 
facilities to handle medical emergencies. All 
subjects will receive each dose in the unit 
and will be monitored closely after each 
dose (for approximately 1-2 hours). Each 
subject will receive a phone call 
approximately 12 hours following 
administration of at least the first two doses 
to assess tolerability. They will be provided 
with a thermometer to measure body 
temperature. Thorough monitoring of any 
AEs will be performed throughout the 
treatment period to allow early assessment 
and mitigation of risk.
Only subjects in good general health with a 
diagnosis of mild asthma will participate in 
this study (see Inclusion/Exclusion Criteria, 
Section 5.1 and Section 5.2). Subjects with 
history of severe asthma are excluded from 
the study.

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Potential Risk of Clinical Significance
Summary of Data/Rationale for Risk
Mitigation Strategy
considered to be adverse, nasal irritancy 
[GlaxoSmithKline Document Number 
2010N110564_04].
GSK2245035 has been previously tested in 
subjects with allergic rhinitis with or 
without mild asthma at the same dose (20 
ng) and the same i.n. regimen (8 weekly 
administrations) proposed for this study.
I.n. GSK2245035 induced transient dose 
related increases in body temperature 
(GSK2245035 80 ng maximum 1.37ºC; 
GSK2245035 20 ng group maximum 
0.34ºC). 
I.n. GSK2245035 induced dose related 
increases in the frequency and severity of 
AEs possibly related to cytokine induction. 
Headache was the most frequently reported 
cytokine related AE for all treatments (79% 
subjects in GSK2245035 80 ng, 29% 
subjects in GSK2245035 20 ng and 21% 
subjects in placebo group), including mild, 
moderate and severe reports. 
Fever was a frequently reported cytokine 
related AE at GSK2245035 80 ng (58%) 
while no reports of cytokine related fever 
were noted at GSK2245035 20ng or 

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Potential Risk of Clinical Significance
Summary of Data/Rationale for Risk
Mitigation Strategy
placebo. Cytokine related fever mainly 
emerged after the second dosing visit; most 
cytokine related fever occurred within 12 
hours of dosing. 
One subject receiving GSK2245035 80ng 
was withdrawn due to severe myalgia 
occurring after the second dose. One 
GSK2245035 20ng subject reported myalgia 
and arthralgia of moderate severity. 
At both doses cytokine related AEs were 
self resolving with a median duration of 
approximately ≤ 1 day.
No severe cytokine related events were 
reported at GSK2245035 20ng.
There were no reports of vomiting, diarrhea 
or hypotension in any treatment group.
Nasal irritancy
Nasal irritancy was noted in rats and 
monkeys after repeat intranasal dosing; the 
effects in the rat were seen only at very high 
dose levels/concentrations (Section 4.5 of 
the Investigator Brochure (IB)) 
[GlaxoSmithKline Document Number 
2010N110564_04].
The finding from the rat and monkey studies 
Thorough monitoring of nasal AEs will be 
performed throughout the treatment period 
to allow early mitigation of risk. Nasal 
examinations will be performed pre-dose 
and 24 hours post-dose on dosing visits 1 
and 8.

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Potential Risk of Clinical Significance
Summary of Data/Rationale for Risk
Mitigation Strategy
is considered to be clinically relevant, but it 
is unlikely to occur in humans at the 
proposed clinical doses; should unexpected 
irritancy be seen, it would be readily 
detectable and has been shown to be readily 
reversible on withdrawal from treatment in 
the monkey.  The no observed adverse 
effect level (NOAEL) in the monkey for 
nasal irritancy (35.9 ng/kg/day) provides 
adequate cover for the proposed 20 ng 
weekly dose in this study.  Therefore, 
GSK2245035 displays an acceptable safety 
profile for an intranasal medicine.
Nasal irritancy has been assessed in clinical 
studies up to 8 weeks’ duration. I.n. 
GSK2245035 is well-tolerated locally as 
determined by nasal examinations, adverse 
events and nasal symptom scores. 
Study Procedures
Anaphylaxis, including bronchospasm 
occurring after bronchial, intranasal, or 
intradermal administration of allergen.
Subjects will inhale (orally and intranasally) 
various concentrations of allergen to which 
they are sensitized. Bronchial allergen 
challenges will be performed at screening, 
and at the follow-up visits (1, 4 and 12 
weeks post final dose).
Intranasal allergen and intradermal testing 
Only sites that have a significant experience 
with administering bronchial, intranasal, and 
intradermal allergen challenges and in the 
management of anaphylaxis and acute 
bronchospasm will be used in this study. 
Careful monitoring of all symptoms will 
take place to assure safety. The responsible 

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Potential Risk of Clinical Significance
Summary of Data/Rationale for Risk
Mitigation Strategy
will be conducted approximately 24 hours 
after the bronchial allergen challenges 
performed at screening and at the 1 and 4 
week follow-up visits.
Allergen testing procedures may induce 
anaphylaxis, including acute bronchospasm.
physician will be appropriately trained to 
treat anaphylaxis and acute bronchospasm, 
including appropriate use of resuscitation 
equipment, which must be close enough to 
respond quickly to an emergency.
Subjects will be continually monitored at 
the study centre for at least 10 hours after 
each bronchial allergen challenge and 6 
hours after intranasal and intradermal 
challenges.  The procedure should be 
stopped immediately if signs/symptoms of 
anaphylaxis and/or acute bronchospam 
develop and rescue medication treatments 
will be readily available.
Subjects will be allowed to leave the unit 
only after the investigator has reviewed and 
assessed the available safety data.
Subjects will receive a Peak Expiratory 
Flow (PEF) meter to measure PEF at home 
from the start of dosing through to 
completion of the 4 week follow-up visit 
assessments. The subject will be instructed 
to contact the investigator if PEF 
measurements are outside the normal range 
for the subject. Subjects will also have their 
PEF levels checked in the unit after 

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Potential Risk of Clinical Significance
Summary of Data/Rationale for Risk
Mitigation Strategy
administration of the final bronchial 
allergen challenge at the 12 week follow-up 
visit.
Sputum induction 
Bronchoconstriction may occur following 
inhalation of saline.
In the event of bronchoconstriction, the 
procedure will be stopped immediately and 
the subject treated with a short-acting 
inhaled bronchodilator.

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4.6.2.
Benefit Assessment
Benefit considerations may include:

Potential benefit of receiving study drug for the study duration that may reduce 
allergic reactivity.

Contributing to the process of developing possible disease-modifying therapies for 
asthma.

Medical evaluations/assessments associated with study procedures [e.g. a physical 
examination, Electrocardiogram (ECG), vital signs, clinical laboratory assessments, 
to screen any potential health issues].
4.6.3.
Overall Benefit: Risk Conclusion
Measures taken to minimise and manage potential risk to subjects participating in this 
study, coupled with the potential benefits justify the conduct of a Phase IIa study with i.n. 
GSK2245035 in mild asthmatics.
5.
SELECTION OF STUDY POPULATION AND 
WITHDRAWAL CRITERIA
Specific information regarding warnings, precautions, contraindications, adverse events, 
and other pertinent information on the GSK investigational product or other study 
treatment that may impact subject eligibility is provided in the IB [GlaxoSmithKline 
Document Number 2010N110564_04].
Deviations from inclusion and exclusion criteria are not allowed because they can 
potentially jeopardize the scientific integrity of the study, regulatory acceptability or 
subject safety. Therefore, adherence to the criteria as specified in the protocol is essential.
5.1.
Inclusion Criteria
A subject will be eligible for inclusion in this study only if all of the following criteria 
apply:
AGE
1. Between 18 and 65 years of age inclusive, at the time of signing the informed 
consent.
TYPE OF SUBJECT AND DIAGNOSIS INCLUDING DISEASE SEVERITY
2. Diagnosis of asthma, as defined by the history of respiratory symptoms such as 
wheeze, shortness of breath, chest tightness and cough that vary over time and in 
intensity, together with variable expiratory airflow limitation [GINA guideline, 2015] 
for at least 6 months prior to screening.
3.
Current asthma therapy. Intermittent short-acting beta-agonist (SABA) alone (on 

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average for no more than 2 days per week)
4. Positive skin prick test (3mm or more greater than negative control) to common 
perennial or seasonal aeroallergen(s) (i.e., house dust mite, cat dander, grass pollen) 
at screening.
5. Pre-bronchodilator FEV1 >70 % predicted normal at Screening Visit 1
NOTE: Predicted values will be based upon Quanjer, 2012.
6. EAR with 20 % FEV1 decrease between 5 and 30 minutes after the final allergen 
concentration is obtained at the screening BAC (decreases relative to the saline).
7. LAR with three FEV1 decreases of 15 % between 4 and 10 hours after the final 
allergen concentration is obtained at the screening bronchial allergen challenge, with 
two FEV1 decreases being at consecutive time points (decreases relative to the 
saline).
8. Subjects who are current non-smokers (defined as no use of any tobacco products in 
the 6-month period preceding the screening visit) and have a pack history of < 10 
pack years
Number of pack years = (number of cigarettes per day/20) x number of years smoked
WEIGHT
9. Bodyweight ≥ 45kg
SEX
10. Male OR female of non-reproductive potential
Males:
Male subjects with female partners of child bearing potential must comply with the  
contraception requirements in Appendix 5 (Modified List of Highly Effective Methods 
for Avoiding Pregnancy) from the time of first dose of study medication until the final 
follow-up visit.
This is an all-inclusive list of those methods that meet the following GSK definition of 
highly effective: having a failure rate of less than 1% per year when used consistently and 
correctly and, when applicable, in accordance with the product label. For non-product 
methods (e.g., male sterility), the investigator determines what is consistent and correct 
use. The GSK definition is based on the definition provided by the International 
Conference on Harmonization of Technical Requirements for Registration of 
Pharmaceuticals for Human Use (ICH) [ICH M3 (R2), 2009].”
The investigator is responsible for ensuring that subjects understand how to properly use 
these methods of contraception.

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Females: 
A female subject is eligible to participate if she is not pregnant (as confirmed by a 
negative [serum or urine] human chorionic gonadotrophin (hCG) test), not lactating, and 
where the following condition applies:
Non-reproductive potential defined as:
Pre-menopausal females with one of the following:
Documented tubal ligation
Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion
Hysterectomy
Documented Bilateral Oophorectomy
Postmenopausal defined as 12 months of spontaneous amenorrhea [in 
questionable cases a blood sample with simultaneous follicle stimulating 
hormone (FSH) and estradiol levels consistent with menopause (refer to 
laboratory reference ranges for confirmatory levels)].  Females on hormone 
replacement therapy (HRT) and whose menopausal status is in doubt will be 
required to use one of the highly effective contraception methods if they wish to 
continue their HRT during the study.  Otherwise, they must discontinue HRT to 
allow confirmation of post-menopausal status prior to study enrolment.
INFORMED CONSENT
11. Capable of giving signed informed consent which includes compliance with the 
requirements and restrictions listed in the consent form and in this protocol.

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5.2.
Exclusion Criteria
A subject will not be eligible for inclusion in this study if any of the following criteria 
apply:
CONCURRENT CONDITIONS/MEDICAL HISTORY (INCLUDES LIVER 
FUNCTION AND QTc INTERVAL) 
1. Alanine transaminase (ALT) >2xUpper Limit of Normal (ULN) and bilirubin 
>1.5xULN (isolated bilirubin >1.5xULN is acceptable if bilirubin is fractionated and 
direct bilirubin <35%).
2. Current or chronic history of liver disease, or known hepatic or biliary abnormalities 
(with the exception of Gilbert's syndrome or asymptomatic gallstones)
3. Heart rate corrected QT interval (QTc) > 450 msec or QTc > 480 msec in subjects 
with Bundle Branch Block
NOTES:

The QTc is the QT interval corrected for heart rate according to Bazett’s 
formula (QTcB), Fridericia’s formula (QTcF), or another method, machine-read 
or manually over-read.

The specific formula that will be used to determine eligibility and 
discontinuation for an individual subject should be determined prior to initiation 
of the study. In other words, several different formulae cannot be used to 
calculate the QTc for an individual subject and then the lowest QTc value used 
to include or discontinue the subject from the trial.

For purposes of data analysis, QTcB, QTcF, another QT correction formula, or a 
composite of available values of QTc will be used as specified in the Reporting 
and Analysis Plan (RAP).
4. Asthma exacerbation requiring treatment with oral corticosteroids or hospitalization 
within 3 months prior to screening
5. History of life-threatening asthma, defined as an asthma episode that required 
intubation and/or was associated with hypercapnea, respiratory arrest or hypoxic 
seizures within the last 10 years.
6. Evidence of concurrent respiratory diseases such as pneumonia, pneumothroax, 
atelectasis, pulmonary fibrotic disease, allergic bronchopulmonary aspergillosis, 
cystic fibrosis, bronchopulmonary dysplasia, or other respiratory abnormalities other 
than asthma.
7. Other concurrent diseases/abnormalities: A subject must not have any clinically 
significant, uncontrolled condition or disease state that, in the opinion of the 
investigator, would put the safety of the subject at risk through study participation or 
would confound the interpretation of the efficacy results if the condition/disease 
exacerbated during the study.
8. Respiratory tract infection that is not resolved within 2 weeks prior to screening.

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CONCOMITANT MEDICATIONS
9. Treatment with intranasal steroid, inhaled corticosteroid (ICS) with or without long-
acting beta2-agonist (LABA), and treatment with non-ICS controller asthma 
medications (i.e., leukotriene modifier, methylxanthines) within 4 weeks prior to 
screening.
10. Use of long-acting antihistamines within 7 days’ or short-acting antihistamines within 
72 hours prior to the screening skin prick test.
11. Treatment with systemic corticosteroids within 6 weeks prior to screening.
12. Use of inhaled SABAs as rescue treatment on average for more than 2 days per week.
RELEVANT HABITS
13. History of regular alcohol consumption within 6 months of the study defined as: 
An average weekly intake of >14 units for males and females.  One unit is 
equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 ml) of wine or 
1 (25 ml) measure of spirits.
CONTRAINDICATIONS 
14. Patient known to be intolerant to salbutamol or albuterol
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.
DIAGNOSTIC ASSESSMENTS AND OTHER CRITERIA
16. Presence of hepatitis B surface antigen (HBsAg), positive hepatitis C antibody test 
result at screening or within 3 months prior to first dose of study treatment.  For 
potent immunosuppressive agents, subjects with presence of hepatitis B core antibody 
(HBcAb) should also be excluded.
17. A positive pre-study drug/alcohol screen
18. A positive test for Human Immunodeficiency Virus (HIV) antibody
19. Where participation in the study would result in donation of blood or blood products 
in excess of 500 mL within a 56 day period.
20. 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)
21. Exposure to more than 4 investigational medicinal products within 12 months prior to 
the first dosing day

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5.3.
Screening Failures/Re-screening
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT) 
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.6).
Re-screening will be permitted in this study, in discussion with the Medical Monitor. 
Sites are encouraged to screen subjects who have previously undergone BAC and have 
demonstrated a LAR. 
Re-screened subjects should be assigned a separate number from their initial screening 
number and complete a new informed consent form. 
5.4.
Withdrawal/Stopping Criteria
Reasons for withdrawal from the study may include:

Use of a prohibited medication that cannot be safely discontinued while maintaining 
the subject’s health.

Treatment code is unblinded.

Pregnancy: Any female subject who becomes pregnant.
The following actions must be taken in relation to a subject who fails to attend the clinic 
for a required study visit:

The site must attempt to contact the subject and re-schedule the missed visit as soon 
as possible. 

The site must counsel the subject on the importance of maintaining the assigned visit 
schedule and ascertain whether or not the subject wishes to and/or should continue in 
the study. 

In cases where the subject is deemed ‘lost to follow up’, the investigator or designee 
must make every effort to regain contact with the subject (where possible, 3 
telephone calls and if necessary a certified letter to the subject’s last known mailing 
address or local equivalent methods). These contact attempts should be documented 
in the subject’s medical record. 

Should the subject continue to be unreachable, only then will he/she be considered to 
have withdrawn from the study with a primary reason of “Lost to Follow-up”. 
A subject may withdraw from study treatment at any time at his/her own request, or may 
be withdrawn at any time at the discretion of the investigator for safety, behavioural or 
administrative reasons. If a subject withdraws from the study, he/she may request 
destruction of any samples taken, and the investigator must document this in the site 
study records.

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Subjects withdrawn from the study while already exposed to IP will be asked to complete 
all safety assessments as outlined in the Time and Events Table (Section 7.1.2). Relevant 
pages of the case report form (CRF) should be completed by the investigator.
The replacement of withdrawn subjects will be at the discretion of the sponsor. If a 
subject misses one treatment dose, the investigator will consult with the GSK medical 
monitor as to whether the subject can receive further treatment. If more than one 
treatment dose is missed, then the subject may be withdrawn from the study, at the 
discretion of the Sponsor, in consultation with the investigator.
If subjects prematurely discontinue the study, additional replacement subjects may be 
recruited and assigned the next available randomisation number, at the discretion of the 
Sponsor, in consultation with the investigator.
5.4.1.
Liver Chemistry Stopping Criteria
Liver chemistry stopping and increased monitoring criteria have been designed to 
assure subject safety and evaluate liver event etiology (in alignment with the Food and 
Drug Administration (FDA) premarketing clinical liver safety guidance). 
http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guid
ances/UCM174090.pdf
Liver Safety Required Actions and Follow up Assessments Section can be found in 
Appendix 2.
5.4.1.1.
Study Treatment Restart or Rechallenge
Study treatment restart or rechallenge after liver chemistry stopping criteria are met by 
any subject participating in this study is not allowed.
5.4.2.
QTc Stopping Criteria
The same QT correction formula must be used for each individual subject to 
determine eligibility for and discontinuation from the study.  This formula may 
not be changed or substituted once the subject has been enrolled.

For example, if a subject is eligible for the protocol based on QTcB, then QTcB 
must be used for discontinuation of this individual subject as well.

Once the QT correction formula has been chosen for a subject’s eligibility, the 
same formula must continue to be used for that subject for all QTc data being 
collected for data analysis.  Safety ECGs and other non-protocol specified 
ECGs are an exception.  
The QTc should be based on single or averaged QTc values of triplicate 
electrocardiograms obtained over a brief (e.g., 5-10 minute) recording period.  
A subject who meets either of the bulleted criteria below will be withdrawn from the 
study:

QTc > 500 msec OR Uncorrected QT > 600 msec

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
Change from baseline of QTc > 60 msec
For subjects with underlying bundle branch block, follow the discontinuation criteria 
listed below:
Baseline QTc with Bundle Branch Block
Discontinuation QTc with Bundle 
Branch Block
< 450 msec
> 500 msec
450 – 480 msec
≥ 530 msec
5.5.
Subject and Study Completion
A completed subject is one who has completed all phases of the study including all of the 
follow-up visits.
The date of the end of the study is defined as the date of the last subject’s last follow-up 
visit.
6.
STUDY TREATMENT
6.1.
Investigational Product and Other Study Treatment
The term ‘study treatment’ is used throughout the protocol to describe any combination 
of products received by the subject as per the protocol design. Study treatment may 
therefore refer to the individual study treatments or the combination of those study 
treatments.

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Study Treatment
Product name:
GSK2245035 Nasal Spray 
Solution 0.2 mcg/mL
Placebo
Formulation 
description:
A solution formulation in saline 
preserved with Benzalkonium 
Chloride and Disodium Edetate
As for GSK2245035 nasal spray 
solution except for omission of 
the active ingredient
Dosage form:
Nasal spray solution
Nasal spray solution
Unit dose 
strength(s)/Dosage 
level(s):
10 ng per actuation
Not applicable
Route of 
Administration:
Intranasal
Once weekly
Intranasal
Once weekly
Dosing 
instructions:
1 spray per nostril
1 spray per nostril
Physical 
description:
An amber glass bottle fitted 
with a screw fit atomizing pump
Visually matched to the active
Device:
Type 1 amber glass bottle fitted 
with a metered Valois VP7 
pump
Type 1 amber glass bottle fitted 
with a metered Valois VP7 pump
Manufacturer/
source of 
procurement:
GSK
GSK
Method for 
individualizing 
dosage:
N/A
N/A
6.2.
Treatment Assignment
Subjects will be assigned to one of two treatments (either 20 ng GSK2245035 or 
placebo), in accordance with the randomization schedule generated by Clinical Statistics, 
prior to the start of the study, using validated internal GSK software RandAll NG.  The 
randomisation will be stratified by centre and also by exposure state during the treatment 
phase to the allergen selected for BAC (two strata – “Presumed” and “Unknown”).  See 
Section 4.4 for further details regarding allergen exposure.
It is planned that an Interactive Response Technology (IRT) will be used in this study to 
assign randomization and drug containers.  It can also be used to emergency unblind 
subjects if required (see Section 6.3).
Once assigned to a subject a randomization number may not be re-used.
More details of how and when a subject is allocated a randomization number and the 
subject numbering convention will be given in the Study Reference Manual (SRM).

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6.3.
Blinding
This will be a double-blind (sponsor open), study (investigators and subjects are blinded, 
and sponsor will be unblinded).
During the study, there will be an unblinded interim analysis. Some key GSK study team 
members may be unblinded as part of this analysis and review. Initially, only the study 
statistician(s) (and supporting programmers), project physician lead, and medical monitor 
will be unblinded, and if further data reviews are required then details of who were un-
blinded to what data and when will be included in the clinical study report.
On-going safety reviews will be conducted during the course of the study in an unblinded 
manner. Key study team members may be unblinded as part of this review. An external 
consultant, who is not a GSK employee, may also be unblinded to enable independent 
review of emerging safety data.
If the GSK medical monitor deems it necessary to unblind data in order to enhance the 
evaluation of safety and tolerability this will be permitted but also documented and 
discussed in the clinical study report.  The GSK medical monitor will also decide whether 
to un-blind the investigator(s)/site(s) at that time, or just the sponsor study staff.  Details 
of who were un-blinded to what data and when would then be included in the clinical 
study report.
Unblinding would occur using the IRT system.
The following will apply. 
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 CRF.
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 CRF.

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GSK’s Global Clinical Safety and Pharmacovigilance (GCSP) staff may unblind the 
treatment assignment for any subject with a Serious Adverse Event (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.
6.4.
Packaging and Labeling
The contents of the label will be in accordance with all applicable regulatory 
requirements.
6.5.
Preparation/Handling/Storage/Accountability
No special preparation of study treatment is required.
One device is dispensed at dosing visit 1 (to use for the first 4 dose administrations), and 
a second device will be dispensed at dosing visit 5 (to use for administration of the fifth 
to eighth doses). Both of these devices will remain in the clinic.  All eight doses will be 
administered to the subjects in the clinic.
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 subjects enrolled in the study may receive study treatment and only authorized site 
staff may supply or administer study treatment.  All study treatments must be stored in a 
secure environmentally controlled and monitored (manual or automated) area in 
accordance with the labelled storage conditions with access limited to the investigator 
and authorized site staff. 
The investigator, institution, or the head of the medical institution (where applicable) is 
responsible for study treatment accountability, reconciliation, and record maintenance 
(i.e. receipt, reconciliation and final disposition records).  
Further guidance and information for final disposition of unused study treatment are 
provided in the SRM. 
Under normal conditions of handling and administration, study treatment is not expected 
to pose significant safety risks to site staff.  Take adequate precautions to avoid direct eye 
or skin contact and the generation of aerosols or mists. In the case of unintentional 
occupational exposure notify the monitor, Medical Monitor and/or GSK study contact.
A Material Safety Data Sheet (MSDS)/equivalent document describing occupational 
hazards and recommended handling precautions either will be provided to the 
investigator, where this is required by local laws, or is available upon request from GSK.

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6.6.
Compliance with Study Treatment Administration
When subjects are dosed at the site, they will receive study treatment directly from the 
investigator or designee, under medical supervision.  The date and time of each dose 
administered in the clinic will be recorded in the source documents.  The dose of study 
treatment and study subject identification will be confirmed at the time of dosing by a 
member of the study site staff other than the person administering the study treatment.  
6.7.
Treatment of Study Treatment Overdose
For this study, any dose of GSK2245035 over the stated maximum of the study treatment 
will be considered an overdose.
GSK does not recommend specific treatment for an overdose. The investigator will use 
clinical judgment to treat any overdose and provide supportive care as appropriate. The 
investigator will inform the GSK Medical Monitor immediately, and will document the 
quantity of the excess dose as well as the duration of the overdosing in the CRF.
Decisions regarding dose interruptions or modifications will be made by the investigator 
in consultation with the Medical Monitor based on the clinical evaluation of the subject.
6.8.
Treatment after the End of the Study
Subjects will not receive any additional treatment from GSK after completion of the 
study because the indication being studied is not life-threatening or seriously debilitating 
and other treatment options are available.
The investigator is responsible for ensuring that consideration has been given to the post-
study care of the subject’s medical condition, whether or not GSK is providing specific 
post-study treatment.
6.9.
Challenge Agents
6.9.1.
Bronchial, intranasal and intra-dermal allergen 
challenge/administration
Appropriate allergen extracts will be purchased by the clinical unit(s) or provided.
The specific allergen extract that will be used for the BAC, NAC and ID challenge will 
be selected on the basis of each subject’s skin prick test. The same allergen will be used 
for each method of testing. The NAC and ID challenge are not required for eligibility.
A detailed description of how all of the allergen extracts will be selected, the preparation 
of the allergen extracts and details of the bronchial, intranasal and intradermal allergen 
challenge techniques can be found in the SRM.

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6.10.
Lifestyle and/or Dietary Restrictions
6.10.1.
Meals and Dietary Restrictions
Dietary restrictions as outlined below will be observed during the time periods specified 
prior to entry to the clinic and dosing.
6.10.2.
Caffeine, Alcohol, and Tobacco

Subjects will abstain from ingesting excessive amounts (more than one cup) of 
caffeine- or xanthine-containing products (e.g. coffee, tea, cola drinks, chocolate) for 
6 hours prior to each dosing. 

During each visit in the clinic, subjects will abstain from alcohol for 24 hours prior 
to entry to the clinic until collection of all assessment is completed.

Subjects will be reminded that all forms of smoking should be avoided during the 
course of the study.
6.10.3.
Activity
Subjects will abstain from strenuous exercise for 48 hours prior to each blood collection 
for clinical laboratory tests.
Subjects should refrain from all recreational drugs from screening until the final follow-
up visit.
6.11.
Concomitant Medications and Non-Drug Therapies
6.11.1.
Permitted Medications and Non-Drug Therapies
Any concomitant medications not specifically permitted will be considered on a case by 
case basis by the investigator and GSK medical monitor. 
Subjects are permitted to take inhaled short-acting β2-agonists (SABA) as rescue 
treatment, but must withhold them for at least 6 hours prior to any study visits with lung 
function assessment or BAC (screening and follow-up visits 1-3).
Medication to control hypertension is permitted, providing it is anticipated to remain 
stable from screening to follow up. Beta blockers are not permitted.
Short term use of anti-histamines is permitted with some limitations as outlined in 
Section 6.11.2; the use of paracetamol and NSAIDS for short-term pain relief is also 
permitted.
6.11.2.
Prohibited Medications and Non-Drug Therapies
Subjects may not receive the following treatments from 4 weeks prior to screening until 
the final follow-up visit:

Intranasal steroids

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
Inhaled corticosteroid (ICS) with or without a long acting beta-2-agonist (LABA)

Non-ICS controller asthma medications (i.e. leukotriene modifier, methylxanthines).
Use of long-acting antihistamines within 7 days’ or short-acting antihistamines within 72 
hours prior to BAC, NAC or ID challenge is prohibited.
Subjects are prohibited from taking systemic corticosteroids within 6 weeks prior to 
screening.
Subjects must abstain from taking prescription or non-prescription drugs (including 
vitamins and dietary or herbal supplements) except as described in Section 6.11.1 within 
7 days prior to the first dose of study medication until completion of the follow-up visits, 
unless in the opinion of the investigator and sponsor the medication will not interfere 
with the study.
7.
STUDY ASSESSMENTS AND PROCEDURES
Protocol waivers or exemptions are not allowed with the exception of immediate safety 
concerns. Therefore, adherence to the study design requirements, including those 
specified in the Time and Events Table, are essential and required for study conduct.
This section lists the procedures and parameters of each planned study assessment.  The 
exact timing of each assessment is listed in the Time and Events Table Section 7.1
The following points must be noted: 

If assessments are scheduled for the same nominal time, THEN the assessments 
should occur in the following order:
1. 12-lead ECG
2. vital signs
3. blood draws.  
Note: The timing of the assessments should allow the blood draw to occur at the 
exact nominal time.

The timing and number of planned study assessments, including safety, 
pharmacokinetic, pharmacodynamic/biomarker assessments may be altered during the 
course of the study based on newly available data to ensure appropriate monitoring. 
Similarly the timing of the post-treatment allergen challenges may change based on 
new information acquired during the course of the study.

Details of the order of planned assessments for each visit and the time windows 
around these visits and assessments will be given in the SRM.

The change in timing or addition of time points for any planned study assessments 
must be documented in a Note to File which is approved by the relevant GSK study 
team member and then archived in the study sponsor and site study files, but this will 
not constitute a protocol amendment. 

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
The Institutional Review Board (IRB)/Independent Ethics Committee (IEC) will be 
informed of any safety issues that require alteration of the safety monitoring scheme
or amendment of the Informed Consent Form. 

No more than 500 mL of blood will be collected over the duration of the study, 
including any extra assessments that may be required.

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7.1.
Time and Events Tables
7.1.1.
Time and Events Table – Screening Visit(s)
Procedure
Screening 
Visit 1 (up to 
Day -28)
Screening Visit 2 
(up to Day -3))
24 hours after 
Screening Visit 2 
(Day -20 to Day -
2)
Notes
There must be a minimum of 48 hours between 
‘Screening Visit 1’ and ‘Screening Visit 2’. This 
ensures adherence to a minimum 3-day window 
between sputum inductions.
There must be a minimum of 48 hours between ‘24 
hours after Screening Visit 2’ and ‘Dosing Visit 1’
Informed consent signing
X
Includes genetics consent
Inclusion and exclusion criteria/eligibility review
X
X
Demographics
X
Medical history (including substance usage and family 
history of premature CV disease)
X
Past and current medical conditions including cardiovascular 
medical history
X
Concomitant medication review
X
Full physical exam including height and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all females)
X
No women of childbearing potential are being 
enrolled. However this test is needed to confirm 
pregnancy status at screening.
Serum FSH and estradiol for post-menopausal females
X
HIV, Hep B and Hep C screen
X
Laboratory assessments (including liver chemistries, 
haematology, clinical chemistry and urinalysis)
X
Screening Spirometry assessment
X
To confirm FEV1 >70% predicted
SAE Collection
X
X
X
12-lead ECG
X
Vital signs
X
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum induction
Saline and incremental BAC challenge plus associated 
spirometry measurements
X
Including pre-procedure FEV1 assessment and 
repeat FEV1 measurements up to 10 hours post BAC

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Procedure
Screening 
Visit 1 (up to 
Day -28)
Screening Visit 2 
(up to Day -3))
24 hours after 
Screening Visit 2 
(Day -20 to Day -
2)
Notes
There must be a minimum of 48 hours between 
‘Screening Visit 1’ and ‘Screening Visit 2’. This 
ensures adherence to a minimum 3-day window 
between sputum inductions.
There must be a minimum of 48 hours between ‘24 
hours after Screening Visit 2’ and ‘Dosing Visit 1’
Sputum induction and processing for BAC associated 
sample collection
X
X
Sputum inductions must be separated by at least 3 
days
Laboratory assessments (haematology only)
X
X
Blood samples should be taken pre-BAC on SV2 
and  on the following day at 24 hrs post BAC
Blood samples for PBMC preparations
X
Blood samples for PBMC preparations (pre-BAC and 
6 hours after BAC at screening visit 2)
Provide diary card, PEF meters, rescue medication and 
thermometers to subjects
X
Only given out to subjects who are eligible following 
the BAC. Thermometers are to be given to record 
temperature as part of capturing AE information.
Nasal lavage
X
Collect nasal lavage to provide a background  
sample. Repeat nasal lavage pre-NAC, then again 5 
minutes and at least 6 hours post-NAC.
Nasal filter collection 
X
Collect nasal filter sample to provide a background 
pre-wash sample, after nasal wash repeat nasal filter 
sampling pre-NAC, then again 5 minutes and at least 
6 hours post-NAC.
NAC
X
Commence after the 24-hour BAC sputum and blood 
biomarker samples have been taken.
Only to be done once subjects have completed a 
successful BAC.
ID challenge
X
ID challenge to be administered after NAC.
Measurements to be made 15 minutes and 6 hours 
post ID challenge. 
Nasal scrape
X
Nasal mucosa sample to be collected at least 
6 hours post-NAC.

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7.1.2.
Time and Events Table – Dosing visit (DV) 1 (Day 1) – DV8 (Week 8) and Early Withdrawal Visit (EW)
Procedure
Pre-dose
Dose
Post-dose
Early-
Withdrawal
Notes
0 hour
20 mins
1 hour
12 hours
24 hours
Outpatient visit
X
X
X
Subjects get discharged 1-2 hours after 
dosing.
Inclusion and 
exclusion 
criteria/eligibility review
X
Only on DV1
Diary card collection 
and review 
X
X
Collect and review diary card at every dosing 
visit 
AE/SAE review
X
X
Concomitant 
medication review
X
X
12-lead ECG
X
X
X
X
Perform on DV1 and DV8 only.
Not required at EW if this occurs during FU
Vital signs (including 
temperature)
X1
X1
X2
X
1 Perform on DV1-8
2 Perform on DV1, 4 and DV8 only.
Not required at EW if this occurs during FU
Laboratory 
assessments (include 
liver chemistries, 
haematology, clinical 
chemistry and 
urinalysis)
X
X
X
Collect samples for laboratory assessments on 
DV1 and DV8,
Not required at EW if this occurs during FU
Blood sample for PK 
analysis
X
X
X
Collect samples on DV1, DV4 and DV8
Genetic sample
X
Take one sample on any of the dosing visits 
after genetics consent.
FeNO measurement
X
Perform FeNO measurements pre-dose on all 
dosing visits 1-8
Nasal examination
X
X
Conducted pre-dose at all study visits and 24 
hours post-dose on DV1, DV4 and DV8

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Procedure
Pre-dose
Dose
Post-dose
Early-
Withdrawal
Notes
0 hour
20 mins
1 hour
12 hours
24 hours
Nasal lavage
X
X
Collect nasal lavage samples pre-dose and 
24hrs post-dose on DV1, DV4 and DV8
Blood sample for 
biomarker analysis
X
X
Collect blood for serum samples and PBMC 
preparations pre-dose and 24 hrs post-dose on 
DV1 and DV8
Blood sample for RNA 
analysis 
X(3)
X(4)
PAX-gene tubes for TLR7/AFFY chip gene 
expression in blood collected pre-dose on 
DV1(3) and 24 hrs after DV8(4)
Call RAMOS
X
Randomisation will occur is at Visit 1 via the 
IRT. Sites should also call RAMOS at DV5.
Dispensing of study 
medication
X
One device will be assigned on DV1. The 
second device will be assigned on DV5.  All 
devices remain in the unit.
Study drug 
administration
X
All doses are administered in the unit.
Sputum induction and 
processing for BAC 
associated sample 
collection
X
Sputum sample is conditional on having 
obtained a sputum sample from the subject at 
screening. Perform sputum induction after 
collection of 24 hour post-dose DV8 blood and 
nasal samples (as pre BAC sample for FUV1).
Follow up phone call
X5
X6
5 Following at least the first two doses of study 
treatment to assess tolerability
6 On DV 2, 3, and 5-7, a follow-up phone call 
will be made approximately 24 hours post-dose 
to check if any CRS events have occurred, and 
if they have, that they have been accurately 
recorded in the diary card and they will then be 
transcribed into the eCRF.
For DV1, 4 and 8 the subjects shall be 
assessed on their return to the unit the 
following day.

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7.1.3.
Time and Events Table – Follow up visits (FUV1 to 3) (Weeks 9 to 20)
Procedure
Follow up Visit 1 
(FUV1)
Follow-up Visit 2 (FUV2)
2 month 
follow-up 
phone 
call
(Week 16)
Follow up Visit 3 (FUV3)
Week 9
(1 week 
after 
DV8)
24 
hours 
later
Week 11
(3 weeks 
after DV8)
Week 12
(4 weeks 
after DV8)
24 
hours 
later
Week 19 
(11 
weeks 
after DV8)
Week 20 
(12 weeks 
after DV8)
24 hours 
later
Notes
AE/SAE review
=========================================
X3
========================
Concomitant 
medication review
=========================================
X3
========================
3 Phone call made to the subject to 
check AEs and concomitant 
medications and confirm availability for 
FUV3
Diarycard collection, 
review and dispensing
X
Review diary card at every visit until 
the end of FUV 2
FeNO measurement
X
X
X
X
X
X
Collect prior to sputum induction
Blood sample for RNA 
analysis
X
PAX tubes for /AFFY chip gene 
expression in blood. Collected prior to 
the bronchial allergen challenge at 
FUV2
Saline and bolus BAC 
challenge plus 
associated spirometry 
measurements
X
X
X
Including pre-procedure FEV1
assessment and repeat FEV1
measurements up to 10 hours post 
BAC
Sputum induction and 
processing for BAC 
associated sample 
collection
X
X
X
X
X
Sputum induction pre-BAC and 24 
hours after BAC
Laboratory 
assessments 
(haematology only)
X
X
X
X
X1
X
Blood samples should be taken pre-
BAC  and  on the following day at 
24hrs post BAC
Blood samples for 
PBMC preparations
X
X
X
Blood samples for PBMC preparations 
(pre-BAC and 6 hours after BAC)

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Procedure
Follow up Visit 1 
(FUV1)
Follow-up Visit 2 (FUV2)
2 month 
follow-up 
phone 
call
(Week 16)
Follow up Visit 3 (FUV3)
Week 9
(1 week 
after 
DV8)
24 
hours 
later
Week 11
(3 weeks 
after DV8)
Week 12
(4 weeks 
after DV8)
24 
hours 
later
Week 19 
(11 
weeks 
after DV8)
Week 20 
(12 weeks 
after DV8)
24 hours 
later
Notes
Nasal lavage
X
X
Collect nasal lavage to provide a
background sample. Repeat nasal 
lavage pre-NAC, then again 5 minutes 
and at least 6 hours post-NAC.
Nasal filter collection
X
X
Collect nasal filter sample to provide a 
background pre-wash sample, after 
nasal wash repeat nasal filter 
sampling pre-NAC, then again 5 
minutes and at least 6 hours post-NA.
NAC
X
X
Commence after the 24-hour BAC 
sputum and blood biomarker  samples 
have been taken at FUV1 and FUV2
ID challenge
X
X
ID challenge to be administered after 
NAC.
Measurements to be made at 15 
minutes and 6 hours post ID 
challenge.
Nasal scrape
X
X
Nasal mucosa sample to be collected 
at least 6 hours post-NAC
Return diary card and 
thermometers
X
Return PEF meters and 
rescue medication
X

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7.2.
Screening and Critical Baseline Assessments
Cardiovascular medical history/risk factors (as detailed in the CRF) will be assessed at 
screening.
The following demographic parameters will be captured: year of birth, sex, race and 
ethnicity.
Medical/medication/family history will be assessed as related to the inclusion/exclusion 
criteria listed in Section 5.  
7.2.1.
Skin prick testing
Seasonal allergen or perennial allergen-specific skin prick tests will be performed at 
screening to confirm eligibility, and to determine the allergen to be used for each subject 
for subsequent BAC, NAC and ID challenge.
7.3.
Efficacy
7.3.1.
Bronchial allergen challenges and spirometry
BAC will be conducted at screening, and at three follow-up visits (see Time and Events 
Tables in Section 7.1). At the Screening Visit incremental doses of allergen will be 
administered and the total dose inducing an EAR and LAR used to calculate a bolus dose 
for use at the 3 FU visits. A detailed description of the BAC technique and assessments 
can be found in the SRM.  All sites participating in the study will be trained in the same 
methodology to ensure consistency.
Following the final FEV1 measurement, subjects will receive rescue salbutamol until the 
FEV1 returns to within 10% of the pre-saline value. If a subject experiences serious 
discomfort the test will be discontinued and rescue medication administered.
7.3.2.
Intranasal and intra-dermal allergen challenge
NAC and ID challenge will be conducted approximately 24 hours after the bronchial 
allergen challenge at screening and at follow-up visits 1 and 2. 
A detailed description of the NAC and ID challenges can be found in the SRM.
In some regions/countries specific challenges agents may not be approved or available 
for use in ID challenge, in which case this assessment shall be omitted.
7.3.3.
Sputum induction
Hypertonic saline induced sputum samples will be collected.  The sputum induction 
process will follow a standard procedure and GSK will ensure all sites are trained prior to 
study start in order to provide consistency.
At screening, the weight of the sputum sample will be measured to confirm that the 
subject is able to produce an adequate induced sputum sample (must be at least 100 mg).

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If a subject did not induce a successful sputum sample at screening then they will not be 
required to try at the follow up visits.
Further information on induction, collection, processing, storage and shipping procedures 
are provided in the SRM.
7.3.4.
TNSS
Four nasal symptoms: nasal congestion, rhinorrhea, nasal itch and sneezing will be 
evaluated on the morning and evening of each day of the study treatment period, from 
DV1 up until follow-up visit 2 (4 weeks after the end of treatment). These will be 
captured on a diary card every morning and evening.  On dosing days, the morning TNSS 
assessment will be captured pre-dose. The subjects will be asked to rate each of these 
symptoms on a 4-point scale:
0 = None:  No symptom whatsoever; Absent
1 = Mild:  Symptom is present, noticeable but not bothersome 
2 = Moderate:  Symptom is bothersome, but tolerable
3 = Severe:  Symptom which are bothersome, harder to tolerate, desiring relief/treatment
The individual symptom scores will be combined to produce a TNSS.
All data will be transcribed from the diary booklet into the CRF.
7.3.5.
Nasal examination
A brief visual nasal examination will be conducted by a trained physician at the time-
points outlined in the Time and Events Tables (Section 7.1). The physician will examine 
the nostrils using appropriate instrumentation (e.g. a small rounded plastic nasal 
speculum and an auroscope fitted with a wide bore examination probe) for any signs of 
irritancy. Any abnormalities will be documented in the CRF.
7.3.6.
Fractional Exhaled Nitric Oxide (FeNO)
FeNO will be measured using a handheld electronic device.  Measurements will be 
obtained in accordance with the [ATS/ERS Recommendations for Standardized 
Procedures for the Online and Offline Measurement of Exhaled Lower Respiratory Nitric 
Oxide and Nasal Nitric Oxide, 2005]. FeNO measurements will be interpreted in 
accordance with the [Official ATS Clinical Practice Guideline: Interpretation of Exhaled 
Nitric Oxide Levels (FeNO) for Clinical Applications, 2011].
Further details will be provided in the SRM.

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7.4.
Safety
Planned time points for all safety assessments are listed in the Time and Events Table
(Section 7.1).  Additional time points for safety tests (such as vital signs, physical exams 
and laboratory safety tests) may be added during the course of the study based on newly 
available data to ensure appropriate safety monitoring.
The Safety Review Team (SRT) is a GSK cross-functional team reviewing all available 
safety data related to the project, including in-stream data from this study, in an ongoing 
manner. The SRT is an internal GSK requirement put in place to ensure holistic 
evaluation of the safety profile of an investigational product with systematic, periodic and 
documented reviews of available safety data, with the appropriate communication and 
escalation of new findings that have the potential to impact patient safety.  
The SRT for this project will review data from this study in collaboration with an 
independent external (to GSK) expert reviewer.
7.4.1.
Adverse Events (AE) and Serious Adverse Events (SAEs)
The definitions of an AE or SAE can be found in Appendix 4.
The investigator and their designees are responsible for detecting, documenting and 
reporting events that meet the definition of an AE or SAE. 
7.4.1.1.
Time period and Frequency for collecting AE and SAE information
Any SAEs assessed as related to study participation (e.g., protocol-mandated procedures, 
invasive tests, or change in existing therapy) or related to a GSK product will be recorded 
from the time a subject consents to participate in the study up to and including the final 
follow-up visit. 
AEs will be collected from the start of Study Treatment until the final follow-up visit (see 
Section 7.4.1.3), at the time-points specified in the Time and Events Table (Section 7.1).
Medical occurrences that begin prior to the start of study treatment but after obtaining 
informed consent may be recorded on the Medical History/Current Medical Conditions 
section of the CRF.
All SAEs will be recorded and reported to GSK within 24 hours, as indicated in 
Appendix 4.
Investigators are not obligated to actively seek AEs or SAEs in former study subjects. 
However, if the investigator learns of any SAE, including a death, at any time after a 
subject has been discharged from the study, and he/she considers the event reasonably 
related to the study treatment or study participation, the investigator must promptly notify 
GSK.

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NOTE: The method of recording, evaluating and assessing causality of AEs and SAEs 
plus procedures for completing and transmitting SAE reports to GSK are provided in 
Appendix 4.
7.4.1.2.
Method of Detecting AEs and SAEs
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended 
and non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrence.  Appropriate questions include:
“How are you feeling?”
“Have you had any (other) medical problems since your last visit/contact?”
“Have you taken any new medicines, other than those provided in this study, since 
your last visit/contact?
7.4.1.3.
Follow-up of AEs and SAEs
After the initial AE/SAE report, the investigator is required to proactively follow each 
subject at subsequent visits/contacts.  All SAEs, and non-serious AEs of special interest 
(as defined in Section 7.4.1.5) will be followed until resolution, until the condition 
stabilizes, until the event is otherwise explained, or until the subject is lost to follow-up 
(as defined in Section 5.4). Further information on follow-up procedures is given in 
Appendix 4.
7.4.1.4.
Cardiovascular and Death Events
Investigators will be required to fill out event specific data collection tools for the 
following AEs and SAEs:
Myocardial infarction/unstable angina
Congestive heart failure
Arrhythmias
Valvulopathy
Pulmonary hypertension
Cerebrovascular events/stroke and transient ischemic attack
Peripheral arterial thrombosis
Deep Venous Thrombosis
Revascularization
For these cardiovascular events detailed above and for all deaths, whether or not they are 
considered SAEs, specific Cardiovascular (CV) and Death sections of the CRF will be
required to be completed. These sections include questions regarding cardiovascular 
(including sudden cardiac death) and non-cardiovascular death.
The CV CRFs are presented as queries in response to reporting of certain CV Medical 
Dictionary for Regulatory Activities (MedDRA) terms. The CV information should be 
recorded in the specific cardiovascular section of the CRF within one week of receipt of a 
CV Event data query prompting its completion.

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The Death CRF is provided immediately after the occurrence or outcome of death is 
reported. Initial and follow-up reports regarding death must be completed within one 
week of when the death is reported.
7.4.1.5.
Disease-Related Events and/or Disease-Related Outcomes Not 
Qualifying as SAEs
The following disease related events (DREs) are common in subjects with mild allergic 
asthma and can be serious/life threatening:

Wheeziness

Chest tightness or chest heaviness

Coughing
Because these events are typically associated with the disease under study, they will not 
be reported according to the standard process for expedited reporting of SAEs to GSK 
(even though the event may meet the definition of a SAE).  These events will be recorded 
on the DRE page in the subject’s CRF within 5 working days.
NOTE: However, if either of the following conditions apply, then the event must be 
recorded and reported as an SAE (instead of a DRE): 

The event is, in the investigator’s opinion, of greater intensity, frequency, or 
duration than expected for the individual subject, or

The investigator considers that there is a reasonable possibility that the event was 
related to treatment with the investigational product.
7.4.1.6.
Regulatory Reporting Requirements for SAEs
Prompt notification by the investigator(s) to GSK of SAEs related to study treatment 
(even for non- interventional post-marketing studies) is essential so that legal obligations 
and ethical responsibilities towards the safety of subjects and the safety of a product 
under clinical investigation are met. 
GSK has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of a product under clinical investigation.  GSK will 
comply with country specific regulatory requirements relating to safety reporting to the 
regulatory authority, Institutional Review Board (IRB)/Independent Ethics Committee 
(IEC) and investigators.
Investigator safety reports are prepared for suspected unexpected serious adverse 
reactions according to local regulatory requirements and GSK policy and are forwarded 
to investigators as necessary.
An investigator who receives an investigator safety report describing a SAE(s) or other 
specific safety information (e.g., summary or listing of SAEs) from GSK will file it with 
the IB and will notify the IRB/IEC, if appropriate according to local requirements.

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7.4.1.7.
Cytokine Related Symptoms (CRS) - Symptoms Grading
In the study, symptoms potentially related to cytokine induction (headache, chills/rigors, 
fever, nausea, vomiting, diarrhoea, arthralgia, myalgia, hypotension) occurring within 24 
hours of dosing will be individually reported as AEs and graded as described in Section 
12.4.2 in Appendix 4. 
7.4.2.
Pregnancy
Details of all pregnancies in female subjects and female partners of male subjects will be 
collected after the start of dosing until the final follow-up visit.
If a pregnancy is reported then the investigator should inform GSK within 2 weeks of 
learning of the pregnancy and should follow the procedures outlined in Appendix 5.
7.4.3.
Physical Exams
A complete physical examination will include, at a minimum, assessment of the 
Cardiovascular, Respiratory, Gastrointestinal and Neurological systems. Height and 
weight will also be measured and recorded. 
A brief physical examination will include, at a minimum assessments of the skin, lungs, 
cardiovascular system, and abdomen (liver and spleen).
Investigators should pay special attention to clinical signs related to previous serious 
illnesses
7.4.4.
Vital Signs
Vital signs will be measured in semi-supine position after 5 minutes rest and will include 
temperature, systolic and diastolic blood pressure, pulse rate and respiratory rate. 
Measurements will be made at screening and various occasions throughout the study as 
outlined in the Time and Events Tables (Section 7.1).  Measurements that deviate 
substantially from normal range, according to the opinion of the Investigator(s), will be 
repeated as soon as possible.
7.4.5.
Electrocardiogram (ECG)
Triplicate 12-lead ECGs will be obtained at screening. Single ECGs will be obtained at 
the other determined time points during the study except if according to the opinion of 
the investigator(s) there are abnormal findings and therefore triplicate ECGs should be 
obtained. These measurements will be obtained at each timepoint during the study using 
an ECG machine that automatically calculates the heart rate and measures PR, QRS, QT, 
and QTc intervals.  Refer to Section 5.4.2 for QTc withdrawal criteria and additional QTc 
readings that may be necessary.

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7.4.6.
Clinical Safety Laboratory Assessments
All protocol required laboratory assessments must be conducted in accordance with the 
Laboratory Manual, and Protocol Time and Events Schedule. Laboratory requisition 
forms must be completed and samples must be clearly labelled with the subject number, 
protocol number, site/centre number, and visit date. Details for the preparation and 
shipment of samples will be provided by the laboratory and are detailed in the laboratory 
manual. Reference ranges for all safety parameters will be provided to the site by the 
laboratory responsible for the assessments.
If additional non-protocol specified laboratory assessments are performed at the 
institution’s local laboratory and result in a change in subject management or are 
considered clinically significant by the investigator (e.g., SAE or AE or dose 
modification) the results must be recorded in the CRF.
Refer to the SRM for appropriate processing and handling of samples to avoid duplicate 
and/or additional blood draws.
All study-required laboratory assessments will be performed by a central laboratory.
NOTE:  Local laboratory results are only required in the event that the central laboratory 
results are not available in time for either a treatment and/or response evaluation to be 
performed. If a local sample is required it is important that the sample for central 
analysis is obtained at the same time. Additionally if the local laboratory results are used 
to make either a treatment or response evaluation, the results must be entered into the 
CRF.
Haematology, clinical chemistry, urinalysis and additional parameters to be tested are 
listed in Table 1.

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Table 1
Protocol Required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Haematology
Platelet Count
RBC Indices:
White Blood Cell (WBC) count 
with Differential:
Red Blood Cell (RBC) 
Count
Mean 
Corpuscular 
Volume 
(MCV)
Neutrophils
Hemoglobin
Mean 
corpuscular 
hemoglobin 
(MCH)
Lymphocytes
Hematocrit
Monocytes
Eosinophils
Basophils
Clinical 
Chemistry 1
Blood Urea 
Nitrogen 
(BUN)
Potassium
AST (SGOT)
Total and direct 
bilirubin
Creatinine
Sodium
ALT (SGPT)
Total Protein
Glucose 
Calcium
Alkaline phosphatise
Albumin
Routine 
Urinalysis

Specific gravity

pH, glucose, protein, blood and ketones by dipstick

Microscopic examination (if blood or protein is abnormal)
Other 
Screening 
Tests

HIV

Hepatitis B (HBsAg)

Hepatitis C (Hep C antibody)

FSH and estradiol (as needed in women of non-child bearing 
potential only)

Alcohol and drug screen (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines)

Urine hCG Pregnancy test 2
NOTES :
1.
Details of Liver Chemistry Stopping Criteria and Required Actions and Follow-Up 
Assessments after liver stopping or monitoring event are given in Section 5.4.1 and 
Appendix 2.
2.
Local urine testing will be standard for the protocol unless serum testing is required 
by local regulation or ethics committee.
All laboratory tests with values that are considered clinically significantly abnormal 
during participation in the study should be repeated until the values return to normal or 
baseline.  If such values do not return to normal within a period judged reasonable by the 
investigator, the etiology should be identified and the sponsor notified.

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7.4.7.
PEF monitoring
Subjects will be given a PEF peak flow meter at screening, along with a diary card. This 
will be used for safety monitoring after the BACS. Subjects will record their PEF 
readings each morning and evening onto the diary card every day from the start of study 
treatment on DV 1 until the 4 week follow-up visit.  
The subject will be instructed to contact the investigator if at any time during the study 
the PEF measurements go outside the normal range for the subject. These instructions 
will be clearly printed on the diary card.
All information from the diary card will be transcribed into the eCRF.
7.5.
Pharmacokinetics
7.5.1.
Blood Sample Collection
Blood samples for pharmacokinetic (PK) analysis of GSK2245035 will be collected at 
the time points indicated in Section 7.1, Time and Events Tables.  The actual date and 
time of each blood sample collection will be recorded.  The timing of PK samples may be 
altered and/or PK samples may be obtained at additional time points to ensure thorough 
PK monitoring.
Processing, storage and shipping procedures are provided in the SRM.
7.5.2.
Sample Analysis
Plasma analysis will be performed under the control of Platform Technology Sciences 
(PTS)-DMPK/Scinovo, GlaxoSmithKline, the details of which will be included in the 
Study Reference Manual (SRM). Concentrations of GSK2245035 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).
Once the plasma has been analyzed for GSK2245035, any remaining plasma may be 
analyzed for other compound-related metabolites and the results reported under a 
separate PTS-DMPK/Scinovo, GlaxoSmithKline protocol.
7.6.
Exploratory Biomarkers
7.6.1.
Pharmacodynamic Markers
Details for the collection of samples, processing, storage and shipping procedures for all 
samples are provided in the SRM. Time-points for all sample collections are shown in the 
Time and Events Tables (Section 7.1).
7.6.1.1.
PD blood samples
Blood samples will be collected before and 24 hours post treatment on DV 1 and 8 for 
serum to measure blood PD biomarkers. 

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Blood will be collected before and 24 hours after treatment on DV 1 and 8 and prior to 
bronchial allergen challenges for preparation of peripheral blood mononuclear cells 
(PBMC). These samples may be used for CHIP cytometry for cellular subset 
phenotyping. If this biomarker analysis is performed it may be reported separately.
7.6.1.2.
Blood PAX collection
Blood PAX samples for AFFYCHIP gene expression (mRNA transcriptome) will be 
collected before dosing on DV 1, 24 hours after DV 8 and prior to BAC on FUV2.
7.6.1.3.
PD nasal lavage samples
Nasal lavage samples to measure PD biomarkers will be collected before and 24 hours 
post treatment on DV1, DV4 and DV8.
7.6.1.4.
Bronchial allergen challenge (BAC) associated biomarkers in induced 
sputum samples
Sputum samples will be collected at the time points indicated in the time and events 
tables (see Section 7.1). Sputum sample collection at dosing/follow up visits is 
conditional on having obtained a sputum sample from the subject at screening.
Detailed information on collection, processing, storage and shipping procedures are 
provided in the SRM.
7.6.1.5.
Bronchial allergen challenge (BAC) associated biomarkers in blood 
samples
Blood samples will be collected before, 6 hours and 24 hours after each BAC:

For haematology, (WBC with differential) for blood eosinophil counts (before and 
24 hours after BAC)

To prepare PBMC that may be used for CHIP cytometry (before and 6 hours after 
BAC).  If this biomarker analysis is performed it may be reported separately.
7.6.1.6.
Nasal allergen challenge associated biomarkers in nasal fluid samples
Nasal filter and nasal lavage samples to measure exploratory biomarkers will be collected 
at the timepoints indicated in the Time and Events Tables, Section 7.1.
7.6.1.7.
Cellular profile of nasal mucosa
Nasal scrapes may be collected after NAC following the completion of the collection of 
the 6 hour nasal lavage and nasal filter samples (at screening and at FUV1 and FUV2). 
Samples of suitable quality may be used for cellular analysis for example by CHIP 
cytometry or RNA-sequencing of single cells (if subjects have given informed consent 
for genetic analysis techniques).  If this biomarker analysis is performed it may be 
reported separately.

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7.6.2.
Novel Biomarkers
With the subject’s consent, nasal tissue, nasal fluid, sputum and blood samples will be 
collected during this study and may be used for the purposes of measuring novel 
biomarkers to identify factors that may influence the disease for study treatment, and/or 
medically related conditions, as well as the biological and clinical responses to 
GSK2245035.  If relevant, this approach will be extended to include the identification of 
biomarkers associated with adverse events.
Samples will be collected at the time points indicated in the Time and Events Table 
Section 7.1. The timing of the collections may be adjusted on the basis of emerging 
pharmacokinetic or pharmacodynamic (PD) data from this study or other new 
information in order to ensure optimal evaluation of the PD endpoints.
Novel candidate biomarkers and subsequently discovered biomarkers of the biological 
response associated with asthma or medically related conditions and/or the action of 
GSK2245035 may be identified by application of:

Measurement of the levels of a subset of RNA species on blood and sputum samples.

Proteome analysis of plasma, nasal and sputum samples.
All samples will be retained for a maximum of 15 years after the last subject completes 
the trial. 
7.7.
Genetics
Information regarding genetic research is included in Appendix 3.
8.
DATA MANAGEMENT

For this study, subject data will be entered into GSK defined CRFs, transmitted 
electronically to GSK or designee and combined with data provided from other 
sources in a validated data system.

Paper diary subject data will be collected and transcribed into GSK defined CRFs by 
the site personnel.

Management of clinical data will be performed in accordance with applicable GSK 
standards and data cleaning procedures to ensure the integrity of the data, e.g., 
removing errors and inconsistencies in the data.

Adverse events and concomitant medications terms will be coded using MedDRA 
(Medical Dictionary for Regulatory Activities) and an internal validated medication 
dictionary, GSKDrug. 

CRFs (including queries and audit trails) will be retained by GSK, and copies will be 
sent to the investigator to maintain as the investigator copy.  Subject initials will not 
be collected or transmitted to GSK according to GSK policy.

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9.
STATISTICAL CONSIDERATIONS AND DATA 
ANALYSES
9.1.
Hypotheses
No formal hypotheses will be tested. The primary study objective will assess the effect of 
a weekly dose of 20ng GSK2245035 vs Placebo by precision estimation and credible 
intervals to estimate the magnitude of underlying effect sizes.  A Bayesian approach will 
be utilised to obtain the posterior distribution for the mean treatment effects for the 
endpoints in the study (e.g. posterior distributions for percentage attenuation for the 
endpoint LAR:  minimum FEV1 between 4-10 hours following allergen challenge one 
week after treatment.). 
Other objectives will assess other pharmacodynamic effects and safety and tolerability of 
GSK 2245035.
9.2.
Sample Size Considerations
The following was performed for each of the proposed LAR primary endpoints:

Success criteria: declare endpoint success if the posterior probability of any 
treatment attenuation exceeds 0.7.

The probability of declaring a success has been computed for a range of true percent 
attenuations and sample sizes (n per arm equal allocation). To make the figures 
easier to interpret the “nuisance” parameters of the placebo response and the 
standard deviation have been accounted for as follows:

Eight previous GSK BAC studies (Crossover design) have been re-analysed to obtain
estimates of the average placebo response and the associated estimate of residual 
variability applicable to the proposed Parallel Group design.

These estimates were combined in a random effects meta-analysis to obtain posterior 
distributions for the underlying parameter values (placebo average response, residual 
variability (for an individual subject) and a between study variance component). The 
future placebo response was computed as the sum of random draws from the 
posterior distributions of the placebo average response parameter and a normal 
distribution with mean zero and variance equal to the between study variance 
parameter. The future GSK2245035 response was derived using the simulation 
scenario’s true percentage attenuation. A pooled sample variance value was 
generated using a Chi-Square distribution with degrees of freedom appropriate to the 
simulation scenario and a sampled variance from the meta-analysis posterior 
distribution. Those numbers were entered into the standard formulae for power 
curves. Repeating this process 10,000 times and taking the median of the resulting 
power values constructs “Assurance” operational characteristics for the combination 
of true percent attenuation and sample size under evaluation.

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
These assurance operational curves have been summarised graphically. The x-axis is 
a hypothetical true drug effect that is assumed to be known for experimental design 
purposes.

The simulations can also be used to compute the minimum observed percentage 
attenuations that would result in a successful study outcome. The median (of the 
10,000 simulations) have been derived for a variety of sample sizes and success 
criteria (probability thresholds 0.7, 0.75, 0.8, 0.85, 0.95) and displayed graphically. 
The y-axis is the observed effect from the study data.
Endpoint: LAR:  minimum FEV1 between 4-10 hours following allergen challenge

Figure 1 illustrates the operating characteristics for the proposed design (n=20 
completers per arm) that have a reasonable chance of meeting the definition of 
success. 

The best guess for the true treatment effect comes from the AZ study with AZD8848 
in subjects with mild asthma, which dosed out of the allergen season and observed an 
approximately 18% effect size (Figure 2) [Leaker , 2012].

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Figure 1
LAR:  minimum FEV1 between 4-10 hours following allergen 
challenge: Probability of Success (Assurance) with Success 
threshold of 0.7 (akin to a risk of false positives of 30%)

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Figure 2
LAR:  minimum FEV1 between 4-10 hours following allergen 
challenge: Minimum observed % attenuations needed to declare 
success for a variety of sample sizes and Success thresholds 
(levels of false positive risk)

Endpoint: LAR:  weighted mean FEV1 between 4-10 hours following allergen 
challenge
Figure 3 and Figure 4 are analogous to Figure 1 and Figure 2. The corresponding AZ 
point estimate was 27% attenuation. Similar conclusions can be drawn.

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Figure 3
LAR:  weighted mean FEV1 between 4-10 hours following allergen 
challenge: Probability of Success (Assurance) with Success 
threshold of 0.7 (akin to a risk of false positives of 30%)

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Figure 4
LAR:  weighted mean FEV1 between 4-10 hours following allergen 
challenge: Minimum observed % attenuations needed to declare 
success for a variety of sample sizes and Success thresholds 
(levels of false positive risk)
9.2.1.
Sample Size Assumptions
The sample size assumes that the behaviour of the placebo response and the variability of 
the endpoints in this study are similar to the eight historical GSK studies (Table 2).

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Table 2
Input data for Sample Size Work
LAR Minimum FEV1 between 4-10h
LAR Weighted Mean FEV1 between 
4-10h
GSK 
Internal 
Study ID
Average 
Placebo 
Response
Degrees of 
Freedom
Residual 
Variability
Average 
Placebo 
Response
Degrees of 
Freedom
Residual 
Variability
-1.3975
33.5
0.40571
-0.9462
33.1
0.30417
-1.0864
45.9
0.13063
-0.7093
42.3
0.11430
-1.1238
22.9
0.30521
-0.6916
22.8
0.18820
-0.9006
48.9
0.07344
-0.3749
47.9
0.02191
-0.7309
85.3
0.10717
-0.4662
84.4
0.07528
-0.9027
31.9
0.27740
-0.5502
34.0
0.20925
-1.4484
20.4
0.40528
-0.9817
20.3
0.31342
-0.8129
69.6
0.15866
-0.5260
71.8
0.08695
Assuming the true treatment effect of GSK2245035 is no worse than the results from the 
AstraZeneca study and 20 subjects complete follow up assessments the probability of 
meeting the success criteria for Min LAR and WM LAR are approximately 78% and 81% 
respectively. This is considered sufficient justification for requiring 20 completers.
9.2.2.
Sample Size Sensitivity
Probability of success (Assurance) was determined for all combinations of these three 
factors: success thresholds of 0.95, 0.85, 0.8, 0.75 and 0.7, sample sizes (per arm) of 5, 
10, 15, 20, 25, 30, 35, 40, 45, 50, 100 and Known % attenuations of 0% to 30% by 2% 
increments, 35 to 50% by 5% increments and 60% to 100% by 10% increments. Full 
results are available on file.
9.2.3.
Sample Size Re-estimation or Adjustment
No sample size re-estimation is currently planned for this study beyond that described in 
Section 9.3.2.  However, if during the course of the study, new information becomes 
available about clinically meaningful differences or variability estimates, a sample size 
re-estimation may be conducted.
9.3.
Data Analysis Considerations
9.3.1.
Analysis Populations
Subjects who receive at least one dose of study medication will be included in an ‘All 
Subjects’ population.  Furthermore, subjects in the ‘All Subjects’ population who have a 
successful screening sputum sample will be included in the ‘Sputum Producers’ 
population.
PPD

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Additionally, subjects from the All Subjects population who were not on Placebo and 
who provide GSK2245035 pharmacokinetic samples will be included in a ‘PK’ 
population. 
Other populations, such as a Per-protocol population, may be defined in the RAP.
9.3.2.
Interim Analysis
Recruitment into the study would continue whilst the interim analysis is taking place.
The interim analysis will require knowledge of which subjects are in the Placebo arm and 
determine how many of those Placebo subjects failed to meet the screening inclusion 
criteria for LAR in their 1 week follow up visit (i.e. did not show three FEV1 decreases of 
15 % relative to saline baseline between 4 and 10 hours, with two FEV1 decreases 
being at consecutive time points at 1 week follow up).  The number of subjects on 
placebo who do not exhibit an LAR (non-LAR responders) will be determined from the 
available interim data.  This number will be checked against Table 3 to advise whether 
the study should continue or whether further review of the interim analysis data is 
required to assess the impact. For example, if at the time the interim analysis occurs 10 
subjects were on Placebo and 4 of those 10 did not demonstrate an LAR response then 
the interim analysis would recommend reviewing the data further.

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Table 3
Lookup table for Interim Analysis Decision Pathway
Number of non-LAR responders on Placebo
0
1
2
3
4
5
6
7
8
9 10 11 12 13 14 15 16 17 18 19 20
Number of Placebo At Interim
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
Continue Study
Data Review
NA
Assessing impact of non-LAR response rate: To evaluate whether recruiting additional 
subjects may counteract the impact of the non-LAR responder alternative version(s) of 
Table 3 would be constructed, but using final sample sizes that range from 20 to 30 
subjects per arm. If any of the resulting estimates for probability of success are within 
10% of the original operating characteristic the study may continue; but requiring 
additional subjects (the smallest number that satisfied the continuation criteria). 
In addition posterior distributions for the percentage attenuation of the primary endpoints 
by GSK2245035 would be constructed from the available interim data. 
Amending protocol / termination of further enrolment (existing subjects would complete 
the study as planned): If the median (point estimate) for the observed treatment effect is 
dramatically different to the assumed effect from the AZ study, the study team may wish 
to predict the probability of success at the end of the study from the interim information 
(details to be provided in the RAP). If this updated probability of success is low then the 
study team may review any of the other available data items at the interim prior to 
amending the protocol / terminating further enrolment (and share these outputs with 
appropriate internal GSK governance boards / senior management). 

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To determine the probability of success in the presence of non-LAR responders (for each 
of the primary endpoints) simulations were performed for each possible combination of 
number of available placebo subjects (n) and number showing no LAR response (r) using 
the AZ point estimates as the true percentage attenuation:
The rate of non-LAR response (p) was assumed to be the same for Placebo and 
GSK2245035 arms. A Beta(0.1, 0.1) prior distribution was assumed for the rate, which 
was updated using conjugate formulae as Beta(0.1 + r, 0.1 + n – r) and a value sampled 
from this posterior distribution. The number of future subjects on placebo with a non-
LAR response was sampled from a Binomial distribution using the sampled value of p 
and (20 – n) observations (the total is a mixture of observed interim data and this sampled 
value). Similarly the number of non-LAR responders on the GSK2245035 arm was 
sampled from a Binomial distribution with the same value of p and 20 observations. Each 
non-responding individual was assigned a constant value for the response, and the 
collection of non-responders were combined with the collection of LAR responders 
(generated in a similar way using the random effects meta-analysis to obtain the expected 
responses). The probability of success was determined for each of the combined datasets 
and the median was derived. If this median was more than 10% smaller than the original 
operating characteristic the recommendation is to further review the data. The 
recommendations from the two endpoints were combined into a single table. Wherever 
there was disagreement between cells the review data decision was chosen.
Sensitivity analyses showed the outcome of this approach is highly dependent on the 
value of the constant used for the non-LAR responses (the further the choice from the 
range of plausible values for LAR responders the more likely the recommendation was to 
review data). The choice of value is arbitrary in nature, but was selected as -0.45 for 
Minimum Change From Saline and -0.2 for Weighted Mean Minimum Change From 
Saline, based upon applying the LAR criteria to seven historical studies and deriving the 
mean responses of all non-LAR responding subjects. Since the decision tables cover all 
interim study possibilities and is pre-specified the consequences of this arbitrary choice 
can be articulated.
9.3.2.1.
Headline results for LAR endpoints at Follow Up 1
The following is conditional on the study not being stopped early by the interim analysis. 
To facilitate internal GSK decision making a data snapshot will be taken when the last 
subject’s BAC FEV1 data (and derived LAR endpoints) from Follow Up 1 are available. 
The associated planned efficacy outputs (to be provided in the RAP) will be produced 
from that (unblinded) snapshot and shared internally with the GSK study team and 
internal GSK governance boards / senior management. This data snapshot will not 
represent a cleaned and locked database and hence its results will not be formally 
reported as they will be superseded by the formal end of study results.

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9.4.
Key Elements of Analysis Plan
9.4.1.
Primary Analyses
Statistical models will be fitted separately to each LAR endpoint (weighted mean and 
minimum FEV1).  These parameters will be analysed using analysis of covariance 
(ANCOVA) with treatment and baseline in the model; other covariates such as centre, 
allergen exposure (‘presumed’/’unknown’) (see Section 6.2) and allergen exposure by 
treatment interaction may be included in the model if the observed data support it (the 
default position is not to include the model terms involving the stratification factor).  
Treatment comparisons of GSK2245035 vs Placebo will be presented with results also 
expressed as percentage attenuations to aid clinical interpretation. The posterior 
probability of any percentage attenuation will be obtained along with appropriate 
summary statistics from the posterior distribution (e.g. 95% and 70% equi-tail Credible 
Intervals).
The principle inferences are expected to come from models with non-informative priors. 
Sensitivity analyses using informative priors based on AZ data may be fitted. 
Full details of the BAC analyses will be provided in the RAP.
9.4.2.
Secondary Analyses
Weighted mean and minimum for EAR will be analysed in a similar manner to that of the 
primary analysis described in Section 9.4.1.
9.4.3.
Safety Analyses
Safety and tolerability data, such as rescue medication, PEF, AEs, will be displayed in 
graphical or summary format or listed as appropriate.  Additional details will be provided 
in the RAP.
9.4.4.
Exploratory Analyses
Pharmacokinetic data will be presented in graphical or tabular form and will be 
summarized descriptively.
Other exploratory analyses will be described in detail in the RAP.
10.
STUDY GOVERNANCE CONSIDERATIONS
10.1.
Posting of Information on Publicly Available Clinical Trial 
Registers
Study information from this protocol will be posted on publicly available clinical trial 
registers before enrollment of subjects begins.

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10.2.
Regulatory and Ethical Considerations, Including the 
Informed Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.
The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent (Subjects who are re-screened are required to 
sign a new ICF)

Investigator reporting requirements (e.g. reporting of AEs/SAEs/protocol deviations 
to IRB/IEC)

GSK will provide full details of the above procedures, either verbally, in writing, or 
both.

Signed informed consent must be obtained for each subject prior to participation in 
the study

The IEC/IRB, and where applicable the regulatory authority, approve the clinical 
protocol and all optional assessments, including genetic research.  

Optional assessments (including those in a separate protocol and/or under separate 
informed consent) and the clinical protocol should be concurrently submitted for 
approval unless regulation requires separate submission.  

Approval of the optional assessments may occur after approval is granted for the 
clinical protocol where required by regulatory authorities.  In this situation, written 
approval of the clinical protocol should state that approval of optional assessments is 
being deferred and the study, with the exception of the optional assessments, can be 
initiated.
10.3.
Quality Control (Study Monitoring)

In accordance with applicable regulations including GCP, and GSK procedures, 
GSK monitors will contact the site prior to the start of the study to review with the 
site staff the protocol, study requirements, and their responsibilities to satisfy 
regulatory, ethical, and GSK requirements.  

When reviewing data collection procedures, the discussion will also include 
identification, agreement and documentation of data items for which the CRF will 
serve as the source document.

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GSK will monitor the study and site activity to verify that the:

Data are authentic, accurate, and complete.

Safety and rights of subjects are being protected.

Study is conducted in accordance with the currently approved protocol and any other 
study agreements, GCP, and all applicable regulatory requirements.
The investigator and the head of the medical institution (where applicable) agrees to 
allow the monitor direct access to all relevant documents.
10.4.
Quality Assurance

To ensure compliance with GCP and all applicable regulatory requirements, GSK 
may conduct a quality assurance assessment and/or audit of the site records, and the 
regulatory agencies may conduct a regulatory inspection at any time during or after 
completion of the study. 

In the event of an assessment, audit or inspection, the investigator (and institution) 
must agree to grant the advisor(s), auditor(s) and inspector(s) direct access to all 
relevant documents and to allocate their time and the time of their staff to discuss the 
conduct of the study, any findings/relevant issues and to implement any corrective 
and/or preventative actions to address any findings/issues identified.
10.5.
Study and Site Closure

Upon completion or premature discontinuation of the study, the GSK monitor will 
conduct site closure activities with the investigator or site staff, as appropriate, in 
accordance with applicable regulations including GCP, and GSK Standard Operating 
Procedures.

GSK reserves the right to temporarily suspend or prematurely discontinue this study 
at any time for reasons including, but not limited to, safety or ethical issues or severe 
non-compliance. For multicenter studies, this can occur at one or more or at all sites.  

If GSK determines such action is needed, GSK will discuss the reasons for taking 
such action with the investigator or the head of the medical institution (where 
applicable).  When feasible, GSK will provide advance notification to the 
investigator or the head of the medical institution, where applicable, of the 
impending action.

If the study is suspended or prematurely discontinued for safety reasons, GSK will 
promptly inform all investigators, heads of the medical institutions (where 
applicable) and/or institution(s) conducting the study. GSK will also promptly 
inform the relevant regulatory authorities of the suspension or premature 
discontinuation of the study and the reason(s) for the action.  

If required by applicable regulations, the investigator or the head of the medical 
institution (where applicable) must inform the IRB/IEC promptly and provide the 
reason for the suspension or premature discontinuation.

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10.6.
Records Retention

Following closure of the study, the investigator or the head of the medical institution 
(where applicable) must maintain all site study records (except for those required by 
local regulations to be maintained elsewhere), in a safe and secure location.  

The records must be maintained to allow easy and timely retrieval, when needed 
(e.g., for a GSK audit or regulatory inspection) and must be available for review in 
conjunction with assessment of the facility, supporting systems, and relevant site 
staff.  

Where permitted by local laws/regulations or institutional policy, some or all of these 
records can be maintained in a format other than hard copy (e.g., microfiche, 
scanned, electronic); however, caution needs to be exercised before such action is 
taken.  

The investigator must ensure that all reproductions are legible and are a true and 
accurate copy of the original and meet accessibility and retrieval standards, including 
re-generating a hard copy, if required.  Furthermore, the investigator must ensure 
there is an acceptable back-up of these reproductions and that an acceptable quality 
control process exists for making these reproductions.

GSK will inform the investigator of the time period for retaining these records to 
comply with all applicable regulatory requirements.  The minimum retention time 
will meet the strictest standard applicable to that site for the study, as dictated by any 
institutional requirements or local laws or regulations, GSK standards/procedures, 
and/or institutional requirements.  

The investigator must notify GSK of any changes in the archival arrangements, 
including, but not limited to, archival at an off-site facility or transfer of ownership 
of the records in the event the investigator is no longer associated with the site.
10.7.
Provision of Study Results to Investigators, Posting of 
Information on Publically Available Clinical Trials Registers 
and Publication
Where required by applicable regulatory requirements, an investigator signatory will be 
identified for the approval of the clinical study report.  The investigator will be provided 
reasonable access to statistical tables, figures, and relevant reports and will have the 
opportunity to review the complete study results at a GSK site or other mutually-
agreeable location.
GSK will also provide the investigator with the full summary of the study results.  The 
investigator is encouraged to share the summary results with the study subjects, as 
appropriate.
GSK will provide the investigator with the randomization codes for their site only after 
completion of the full statistical analysis.
The procedures and timing for public disclosure of the results summary and for 
development of a manuscript for publication will be in accordance with GSK Policy.

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11.
REFERENCES
An Official ATS Clinical Practice Guideline: Interpretation of Exhaled Nitric Oxide 
Levels (FeNO) for Clinical Applications. Am J Respir Crit Care Med. 2011; 184: 
602−615
ATS/ERS Recommendations for Standardized Procedures for the Online and Offline 
Measurement of Exhaled Lower Respiratory Nitric Oxide and Nasal Nitric Oxide. Am J 
Respir Crit Care Med. 2005; 171: 912–930
GlaxoSmithKline Document Number 2010N110564_04. GSK2245035 Investigator’s 
Brochure. December 2015.
GlaxoSmithKline Document Number 2011N120740_00. A randomized, single-blind, 
placebo-controlled first time into human study to investigate the safety, tolerability, 
pharmacokinetics and pharmacodynamics of intranasal dosing with GSK2245035, a 
TLR7 agonist, in healthy volunteers and allergic rhinitics. July 2012.
GlaxoSmithKline Document Number 2011N126275_00. A randomised, double-blind, 
placebo-controlled study to investigate the safety, pharmacodynamics and efficacy 
against allergic reactivity of repeat intranasal administration of the TLR7 agonist 
GSK2245035 in subjects with respiratory allergies. January 2012.
GlaxoSmithKline Document Number 2013N174200_00. A randomized, double blind,
placebo-controlled study to investigate the safety, pharmacodynamics and effect on 
allergic reactivity of repeat intranasal administration of the TLR7 agonist GSK2245035 
in subjects with respiratory allergies. August 2015.
Global Initiative for Asthma (GINA). Global Strategy for Asthma management and 
prevention 2015. Available from www.ginasthma.org.
Hankinson J L, Odencrantz, JR; Feden, KB. Spirometric Reference Values from a 
Sample of the General U.S. Population. AM J Respir Crit care Med. 1999;159:179-187.
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, Policar MS, 
editors. Contraceptive Technology. 20th edition. Atlanta, Georgia: Ardent Media, Inc., 
2011: 50. Table 3-2.
Leaker B, Singh D, Lindgren S, et al., editors. The Effects Of The Novel Toll-Like 
Receptor 7 (TLR7) Agonist AZD8848 On Allergen-Induced Responses In Patients With 
Mild Asthma. American Thoracic Society International Conference; 2012; San Francisco, 
California, USA.
Quanjer PH, Stanojevic S, Cole TJ, Baur X, Hall GL, Culver BH, Enright PL, Hankinson 
JL, Ip MS, Zheng J, Stocks J; ERS Global Lung Function Initiative.  Multi-ethnic 
reference values for spirometry for the 3-95-yr age range: the global lung function 2012 
equations.  Eur Respir J. 2012;40:1324-43.

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12.
APPENDICES
12.1.
Appendix 1 – Abbreviations and Trademarks
Abbreviations
AC
Allergen challenge
AE
Adverse Event
ALT
Alanine aminotransferase (SGPT)
ANOVA
Analysis of Variance
AR
Allergic Rhinitis or Allergic Rhinitics
AST
Aspartate aminotransferase (SGOT)
AUC
Area under concentration-time curve
AUC(0-)
Area under the concentration-time curve from time zero (pre-dose) 
extrapolated to infinite time
%AUCex
Percentage of AUC(0-) obtained by extrapolation
AUC(0-x)
Area under the concentration-time curve from zero (pre-dose) to some 
fixed nominal time x
AUC(0-t)
Area under the concentration-time curve from time zero (pre-dose) to 
last time of quantifiable concentration within a subject across all 
treatments
AUC(0-)
Area under the concentration-time curve over the dosing interval
AZ
AstraZeneca
-HCG
Beta-Human Chorionic Gonadotropin
BAC
Bronchial allergen challenge
BMI
Body mass index
BUN
Blood urea nitrogen
CI
Confidence Interval
CIB
Clinical Investigator’s Brochure
CRF
Case Report Form
CRO
Contract Research Organization
CRS
Cytokine Release Syndrome
CV
Coefficient of variance
DMPK
Drug Metabolism and Pharmacokinetics
DNA
Deoxyribonucleic acid
DRE
Disease Related Event
DV
Dosing Visit
EAR
Early asthmatic response
ECG
Electrocardiogram
eNO
Exhaled Nitric Oxide
FDA
Food and Drug Administration
FEV1
Forced expiratory volume in 1 second
FSH
Follicle Stimulating Hormone
FU
Follow-up
GCP
Good Clinical Practice
GCSP
Global Clinical Safety and Pharmacovigilence

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GLP
Good Laboratory Practice
GSK
GlaxoSmithKline
HBsAg
Hepatitis B surface antigen
hCG
Human chorionic gonadotropin
HIV
Human Immunodeficiency Virus
h/hr
Hour(s)
HR
Heart rate
HRT
Hormone replacement therapy
HVT
Healthy volunteers
IB
Investigator’s Brochure
IC
Intradermal challenge
ICH
International Conference on Harmonization of Technical Requirements 
for Registration of Pharmaceuticals for Human Use
ICS
Inhaled corticosteroids
IEC
Independent Ethics Committee
IFN
Interferon
Ig
Immunoglobulin
I.N.
Intranasal
IND
Investigational New Drug
INR
International normalized ratio
IP
Investigational Product
IP-10
Interferon inducible protein 10
IRB
Institutional Review Board
IRT
Interactive Response Technology
IU
International Unit
IV
Intravenous
Kg
Kilogram
L
Liter
LABA
Long-acting beta2-agonist
LAR
Late asthmatic response
µg
Microgram
µL
Microliter
MCHC
Mean corpuscular hemoglobin concentration
MCV
Mean corpuscular volume
MedDRA
Medical Dictionary for Regulatory Activities
Mg
Milligrams
mL
Milliliter
mm
Millimeters
MSDS
Material Safety Data Sheet
msec
Milliseconds
NAC
Nasal allergen challenge
ng
nanogram
NO
Nitric Oxide
NOAEL
No observed adverse event level
OD
Once a day, once daily
PAX
Paired box gene

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PBMC
Peripheral blood mononuclear cell
PBS
Phosphate buffered saline
PD
Pharmacodynamic
PEF
Peak Expiratory Flow
PK
Pharmacokinetic
PNIF
Peak Nasal Inspiratory Flow
PTS
Platform Technology Sciences
QC
Quality control
QTcB
QT duration corrected for heart rate by Bazett’s formula
QTcF
QT duration corrected for heart rate by Fridericia’s formula
RAP
Reporting and Analysis Plan
RBC
Red blood cells
RNA
Ribonucleic acid
SABA
Short-acting beta2-agonist
SAE
Serious adverse event(s)
SAR
Seasonal allergic rhinitics
SAS
Statistical Analysis Software
SD
Standard deviation
SOP
Standard Operating Procedure
SRM
Study Reference Manual
T
Time of last observed quantifiable concentration
t½
Terminal phase half-life
Th
T helper cell
Tmax
Time of occurrence of Cmax
TNSS
Total nasal symptoms score
TLR7
Toll like receptor 7
Treg
Regulated T cells
ULN
Upper limit of normal
UK
United Kingdom
US
United States
WBC
White blood cells
Trademark Information
Trademarks of the GlaxoSmithKline 
group of companies
Trademarks not owned by the 
GlaxoSmithKline group of companies
NONE
None

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12.2.
Appendix 2: Liver Safety Required Actions and Follow up 
Assessments
Phase II Liver Chemistry Stopping and Increased Monitoring Algorithm
Continue Study Treatment 
Discontinue Study Treatment 
Plus
Bilirubin≥2x
ULN (>35% 
direct) 
or plus
INR>1.5, if 
measured*
Possible
Hy’s Law
ALT≥3xULN
ALT≥5xULN
ALT≥3xULN
Plus
Symptoms of 
liver injury
or 
hypersensitivity
ALT≥3xULN
but able to 
monitor 
weekly for 
4 weeks
No
Yes
Yes
Yes
Yes
No
No
No
No
Yes
ALT≥3xULN
persist for
4 weeks or 
stopping 
criteria 
met
Yes
No
*INR value not applicable to subjects on anticoagulants
Yes
If subject to be monitored weekly must refer to Liver Safety Required  Actions and  Follow  up 
Assessments  section  in  the  Appendix
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*
Phase II liver chemistry increased monitoring criteria with continued therapy
Liver Chemistry Increased Monitoring Criteria – Liver Monitoring Event
Criteria
Actions
ALT 3xULN and <5xULN and bilirubin 
<2xULN, without symptoms believed to 
be related to liver injury or 
hypersensitivity, and who can be 
monitored weekly for 4 weeks

Notify the GSK medical monitor within 24 hours of 
learning of the abnormality to discuss subject safety. 

Subject can continue study treatment

Subject must return weekly for repeat liver 
chemistries (ALT, AST, alkaline phosphatase, 
bilirubin) until they resolve, stabilise or return to 
within baseline 

If at any time subject meets the liver chemistry 
stopping criteria, proceed as described above

If, after 4 weeks of monitoring, ALT <3xULN and 
bilirubin <2xULN, monitor subjects twice monthly 
until liver chemistries normalize or return to within 
baseline.

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References
James LP, Letzig L, Simpson PM, Capparelli E, Roberts DW, Hinson JA, Davern TJ, Lee 
WM. Pharmacokinetics of Acetaminophen-Adduct in Adults with Acetaminophen 
Overdose and Acute Liver Failure. Drug Metab Dispos 2009; 37:1779-1784.

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12.3.
Appendix 3 - Genetic Research
Genetics – Background
Naturally occurring genetic variation may contribute to inter-individual variability in 
response to medicines, as well as an individual's risk of developing specific diseases. 
Genetic factors associated with disease characteristics may also be associated with 
response to therapy, and could help to explain some clinical study outcomes. For 
example, genetic variants associated with age-related macular degeneration (AMD) are 
reported to account for much of the risk for the condition [Gorin, 2012] with certain 
variants reported to influence treatment response [Chen, 2012]. Thus, knowledge of the 
genetic etiology of disease may better inform understanding of disease and the 
development of medicines. Additionally, genetic variability may impact the 
pharmacokinetics (absorption, distribution, metabolism, and elimination), or 
pharmacodynamics (relationship between concentration and pharmacologic effects or the 
time course of pharmacologic effects) of a specific medicine and/or clinical outcomes 
(efficacy and/or safety) observed in a clinical study.
Genetic Research Objectives and Analyses
The objectives of the genetic research are to investigate the relationship between genetic 
variants and: 
Response to medicine, including any treatment regimens under investigation in this study 
or any concomitant medicines;
Asthma susceptibility, severity, and progression and related conditions
Genetic data may be generated while the study is underway or following completion of 
the study.  Genetic evaluations may include focused candidate gene approaches and/or 
examination of a large number of genetic variants throughout the genome (whole genome 
analyses). Genetic analyses will utilize data collected in the study and will be limited to 
understanding the objectives highlighted above.  Analyses may be performed using data 
from multiple clinical studies to investigate these research objectives.
Appropriate descriptive and/or statistical analysis methods will be used. A detailed 
description of any planned analyses will be documented in a Reporting and Analysis Plan 
(RAP) prior to initiation of the analysis. Planned analyses and results of genetic 
investigations will be reported either as part of the clinical RAP and study report, or in a 
separate genetics RAP and report, as appropriate.
Study Population
Any subject who is enrolled in the study can participate in genetic research.  Any subject 
who has received an allogeneic bone marrow transplant must be excluded from the 
genetic research.

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Study Assessments and Procedures
A key component of successful genetic research is the collection of samples during 
clinical studies. Collection of samples, even when no a priori hypothesis has been 
identified, may enable future genetic analyses to be conducted to help understand 
variability in disease and medicine response.  
A 6 ml blood sample will be taken for Deoxyribonucleic acid (DNA) extraction. A Blood 
sample is collected at the baseline visit, after the subject has been randomized and 
provided informed consent for genetic research. Instructions for collection and shipping 
of the genetic sample are described in the laboratory manual. The DNA from the blood 
sample may undergo quality control analyses to confirm the integrity of the sample. If 
there are concerns regarding the quality of the sample, then the sample may be destroyed. 
The blood sample is taken on a single occasion unless a duplicate sample is required due 
to an inability to utilize the original sample.  
The genetic sample is labelled (or “coded”) with the same study specific number used to 
label other samples and data in the study.   This number can be traced or linked back to 
the subject by the investigator or site staff. Coded samples do not carry personal 
identifiers (such as name or social security number). 
Samples will be stored securely and may be kept for up to 15 years after the last subject 
completes the study, or GSK may destroy the samples sooner.  GSK or those working 
with GSK (for example, other researchers) will only use samples collected from the study 
for the purpose stated in this protocol and in the informed consent form.  Samples may be 
used as part of the development of a companion diagnostic to support the GSK medicinal 
product.
Subjects can request their sample to be destroyed at any time.
Informed Consent
Subjects who do not wish to participate in the genetic research may still participate in the 
study.  Genetic informed consent must be obtained prior to any blood being taken.
Subject Withdrawal from Study
If a subject who has consented to participate in genetic research withdraws from the 
clinical study for any reason other than being lost to follow-up, the subject will be given a 
choice of one of the following options concerning the genetic sample, if already 
collected:
Continue to participate in the genetic research in which case the genetic DNA sample is 
retained
Discontinue participation in the genetic research and destroy the genetic DNA sample 
If a subject withdraws consent for genetic research or requests sample destruction for any 
reason, the investigator must complete the appropriate documentation to request sample 

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destruction within the timeframe specified by GSK and maintain the documentation in 
the site study records. 
Genotype data may be generated during the study or after completion of the study and 
may be analyzed during the study or stored for future analysis. 
If a subject withdraws consent for genetic research and genotype data has not been 
analyzed, it will not be analyzed or used for future research.  
Genetic data that has been analyzed at the time of withdrawn consent will continue to be 
stored and used, as appropriate. 
Screen Failures
If a sample for genetic research has been collected and it is determined that the subject 
does not meet the entry criteria for participation in the study, then the investigator should 
instruct the subject that their genetic sample will be destroyed. No forms are required to 
complete this process as it will be completed as part of the consent and sample 
reconciliation process.  In this instance a sample destruction form will not be available to 
include in the site files.
Provision of Study Results and Confidentiality of Subject’s Genetic Data
GSK may summarize the genetic research results in the clinical study report, or 
separately and may publish the results in scientific journals. 
GSK may share genetic research data with other scientists to further scientific 
understanding in alignment with the informed consent. GSK does not inform the subject, 
family members, insurers, or employers of individual genotyping results that are not 
known to be relevant to the subject’s medical care at the time of the study, unless 
required by law. This is due to the fact that the information generated from genetic 
studies is generally preliminary in nature, and therefore the significance and scientific 
validity of the results are undetermined.  Further, data generated in a research laboratory 
may not meet regulatory requirements for inclusion in clinical care.
References
Chen H, Yu KD, Xu GZ. Association between Variant Y402H in Age-Related Macular 
Degeneration (AMD) Susceptibility Gene CFH and Treatment Response of AMD: A 
Meta-Analysis. PloS ONE 2012; 7: e42464
Gorin MB. Genetic insights into age-related macular degeneration: Controversies 
addressing risk, causality, and therapeutics. Mol. Asp. Med. 2012; 33: 467-486.

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12.4.
Appendix 4: Definition of and Procedures for Recording, 
Evaluating, Follow-Up and Reporting of Adverse Events
12.4.1.
Definition of Adverse Events
Adverse Event Definition:

An AE is any untoward medical occurrence in a patient or clinical investigation 
subject, temporally associated with the use of a medicinal product, whether or not 
considered related to the medicinal product.

NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of a medicinal product.
Events meeting AE definition include:

Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (e.g., ECGs, radiological scans, vital signs 
measurements), including those that worsen from baseline, and felt to be clinically 
significant in the medical and scientific judgement of the investigator.

Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition.

New conditions detected or diagnosed after study treatment administration even 
though it may have been present prior to the start of the study.

Signs, symptoms, or the clinical sequelae of a suspected interaction.

Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication (overdose per se will not be reported as an 
AE/SAE unless this is an intentional overdose taken with possible suicidal/self-
harming intent.  This should be reported regardless of sequelae).

"Lack of efficacy" or "failure of expected pharmacological action" per se will not be 
reported as an AE or SAE.  However, the signs and symptoms and/or clinical 
sequelae resulting from lack of efficacy will be reported if they fulfil the definition 
of an AE or SAE.

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Events NOT meeting definition of an AE include:

Any clinically significant abnormal laboratory findings or other abnormal safety 
assessments which are associated with the underlying disease, unless judged by the 
investigator to be more severe than expected for the subject’s condition.

The disease/disorder being studied or expected progression, signs, or symptoms of 
the disease/disorder being studied, unless more severe than expected for the subject’s 
condition.

Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that 
leads to the procedure is an AE.

Situations where an untoward medical occurrence did not occur (social and/or 
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.

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12.4.2.
Definition and Grading of CRS-related Adverse Events
In the study, symptoms potentially related to cytokine induction (headache, chills/rigors, 
fever, nausea, vomiting, diarrhoea, arthralgia, myalgia, hypotension) occurring within 24 
hours of dosing will be individually reported as AEs and graded as described below:

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Cytokine Release Syndrome (CRS) AE grading (continued)
12.4.3.
Definition of Serious Adverse Events
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, 
death due to progression of disease, etc).
Serious Adverse Event (SAE) is defined as any untoward medical occurrence that, 
at any dose:
a.
Results in death
b.
Is life-threatening
NOTE:
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
subject was at risk of death at the time of the event.  It does not refer to an event, which 
hypothetically might have caused death, if it were more severe.
c.
Requires hospitalization or prolongation of existing hospitalization
NOTE:

In general, hospitalization signifies that the subject has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for 
observation and/or treatment that would not have been appropriate in the physician’s 

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office or out-patient setting.  Complications that occur during hospitalization are 
AEs.  If a complication prolongs hospitalization or fulfills any other serious criteria, 
the event is serious.  When in doubt as to whether “hospitalization” occurred or was 
necessary, the AE should be considered serious.

Hospitalization for elective treatment of a pre-existing condition that did not worsen 
from baseline is not considered an AE.
d.
Results in disability/incapacity
NOTE:

The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions.

This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, 
and accidental trauma (e.g. sprained ankle) which may interfere or prevent everyday 
life functions but do not constitute a substantial disruption
e.
Is a congenital anomaly/birth defect
f.
Other situations:

Medical or scientific judgment should be exercised in deciding whether reporting is 
appropriate in other situations, such as important medical events that may not be 
immediately life-threatening or result in death or hospitalization but may jeopardize 
the subject or may require medical or surgical intervention to prevent one of the 
other outcomes listed in the above definition.  These should also be considered 
serious.

Examples of such events are invasive or malignant cancers, intensive treatment in an 
emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse
g.
Is associated with liver injury and impaired liver function defined as:

ALT 3xULN and total bilirubin* 2xULN (>35% direct), or

ALT 3xULN and International Normalised Ratio (INR)** > 1.5.
* Serum bilirubin fractionation should be performed if testing is available; if unavailable, 
measure urinary bilirubin via dipstick.  If fractionation is unavailable and ALT 3xULN 
and total bilirubin 2xULN, then the event is still to be reported as an SAE.
** INR testing not required per protocol and the threshold value does not apply to 
subjects receiving anticoagulants.  If INR measurement is obtained, the value is to be 
recorded on the SAE form.

Refer to Appendix 2 for the required liver chemistry follow-up instructions

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12.4.4.
Definition of Cardiovascular Events
Cardiovascular Events (CV) Definition:
Investigators will be required to fill out the specific CV event page of the CRF for the 
following AEs and SAEs:

Myocardial infarction/unstable angina

Congestive heart failure

Arrhythmias

Valvulopathy

Pulmonary hypertension

Cerebrovascular events/stroke and transient ischemic attack

Peripheral arterial thromboembolism

Deep venous thrombosis/pulmonary embolism

Revascularization
12.4.5.
Recording of AEs and SAEs
AEs and SAE Recording:

When an AE/SAE occurs, it is the responsibility of the investigator to review all 
documentation (e.g., hospital progress notes, laboratory, and diagnostics reports) 
relative to the event.

The investigator will then record all relevant information regarding an AE/SAE in 
the CRF

It is not acceptable for the investigator to send photocopies of the subject’s medical 
records to GSK in lieu of completion of the GSK, AE/SAE CRF page.

There may be instances when copies of medical records for certain cases are 
requested by GSK.  In this instance, all subject identifiers, with the exception of the 
subject number, will be blinded on the copies of the medical records prior to 
submission of to GSK.

The investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information.  In such cases, the diagnosis will be 
documented as the AE/SAE and not the individual signs/symptoms.

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12.4.6.
Evaluating AEs and SAEs
Assessment of Intensity
The investigator will make an assessment of intensity for each AE and SAE reported 
during the study and will assign it to one of the following categories:

Mild:  An event that is easily tolerated by the subject, causing minimal discomfort 
and not interfering with everyday activities.

Moderate:  An event that is sufficiently discomforting to interfere with normal 
everyday activities

Severe:  An event that prevents normal everyday activities. - an AE that is assessed 
as severe will not be confused with an SAE. Severity is a category utilized for rating 
the intensity of an event; and both AEs and SAEs can be assessed as severe.

An event is defined as ‘serious’ when it meets at least one of the pre-defined 
outcomes as described in the definition of an SAE.
Assessment of Causality

The investigator is obligated to assess the relationship between study treatment and 
the occurrence of each AE/SAE.

A "reasonable possibility" is meant to convey that there are facts/evidence or 
arguments to suggest a causal relationship, rather than a relationship cannot be ruled 
out.

The investigator will use clinical judgment to determine the relationship.

Alternative causes, such as natural history of the underlying diseases, concomitant 
therapy, other risk factors, and the temporal relationship of the event to the study 
treatment will be considered and investigated.

The investigator will also consult the Investigator Brochure (IB) and/or Product 
Information, for marketed products, in the determination of his/her assessment.

For each AE/SAE the investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality.

There may be situations when an SAE has occurred and the investigator has minimal 
information to include in the initial report to GSK.  However, it is very important 
that the investigator always make an assessment of causality for every event 
prior to the initial transmission of the SAE data to GSK.

The investigator may change his/her opinion of causality in light of follow-up 
information, amending the SAE data collection tool accordingly.

The causality assessment is one of the criteria used when determining regulatory 
reporting requirements.

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Follow-up of AEs and SAEs

The investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as may be indicated or as requested by GSK to 
elucidate as fully as possible the nature and/or causality of the AE or SAE.

The investigator is obligated to assist.  This may include additional laboratory tests 
or investigations, histopathological examinations or consultation with other health 
care professionals.

If a subject dies during participation in the study or during a recognized follow-up 
period, the investigator will provide GSK with a copy of any post-mortem findings, 
including histopathology.

New or updated information will be recorded in the originally completed CRF.

The investigator will submit any updated SAE data to GSK within the designated 
reporting time frames.
12.4.7.
Reporting of SAEs to GSK
SAE reporting to GSK via electronic data collection tool

Primary mechanism for reporting SAEs to GSK will be the electronic data collection 
tool

If the electronic system is unavailable for greater than 24 hours, the site will use the 
paper SAE data collection tool and fax it to the GSK Medical Monitor and GSK 
Study Manager

Site will enter the serious adverse event data into the electronic system as soon as it 
becomes available.

The investigator will be required to confirm review of the SAE causality by ticking 
the ‘reviewed’ box at the bottom of the eCRF page within 72 hours of submission of 
the SAE.

After the study is completed at a given site, the electronic data collection tool (e.g., 
InForm system) 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 subject or receives updated data 
on a previously reported SAE after the electronic data collection tool has been taken 
off-line, the site can report this information on a paper SAE form or to the GSK 
Medical Monitor and GSK Study Manager by telephone.

Contacts for SAE receipt can be found at the beginning of this protocol on the 
Sponsor/Medical Monitor Contact Information page.

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12.5.
Appendix 5: Modified List of Highly Effective Methods for 
Avoiding Pregnancy and Collection of Pregnancy 
Information
12.5.1.
Modified List of Highly Effective Methods for Avoiding Pregnancy
Contraceptive requirements for male subjects with female partners of 
reproductive potential (when applicable).
Male subjects with female partners of child bearing potential must comply with the 
following contraception requirements from the time of first dose of study medication 
until the final follow-up visit.
1.
Vasectomy with documentation of azoospermia. The documentation on male sterility 
can come from the site personnel’s: review of subject’s medical records, medical 
examination and/or semen analysis, or medical history interview.
2.
Male condom plus partner use of one of the contraceptive options below that meets 
the Standard Operating Procedure (SOP) effectiveness criteria including a <1% rate 
of failure per year, as stated in the product label:

Contraceptive subdermal implant

Intrauterine device or intrauterine system

Combined estrogen and progestogen oral contraceptive [Hatcher, 2011] 

Injectable progestogen [Hatcher, 2011]

Contraceptive vaginal ring [Hatcher, 2011]

Percutaneous contraceptive patches [Hatcher, 2011]
These allowed methods of contraception are only effective when used consistently, 
correctly and in accordance with the product label. The investigator is responsible for 
ensuring that subjects understand how to properly use these methods of contraception.
12.5.2.
Collection of Pregnancy Information

Investigator will collect pregnancy information on any female subject, who becomes 
pregnant while participating in this study (this may also be a female partner of a male 
subject in the study)

Information will be recorded on the appropriate form and submitted to GSK within 2 
weeks of learning of a subject's pregnancy.

Subject will be followed to determine the outcome of the pregnancy. The 
investigator will collect follow up information on mother and infant, 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.

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
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.

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 Appendix 4. 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.
Any female subject who becomes pregnant while participatingwill discontinue study 
medication and will be withdrawn from the study

Investigator will attempt to collect pregnancy information on any female partner of a 
male study subject who becomes pregnant while participating in this study.  This 
applies only to subjects who are randomized to receive study medication. 

After obtaining the necessary signed informed consent from the female partner 
directly, the investigator will record pregnancy information on the appropriate form 
and submit it to GSK within 2 weeks 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.
12.5.3.
References
CHMP, 2005 - EMA - Guideline on Adjuvants in Vaccines for Human Use, 
(CHMP/VEG/134716/2004), January 2005.
Cole LA, Khanlian SA, Sutton JM, Davies S, Rayburn WF. Accuracy of home pregnancy 
tests at the time of missed menses. Am J Ob Gyn 2004(190):100-5.
EMA/CHMP/ICH/449035/2009: General principles to address virus and vector shedding. 
http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/
WC500002680.pdf Accessed 28 Nov 2014.
EMEA/CHMP/GTWP/125459/2006: Guideline on the Nonclinical Studies Required 
Before First Clinical Use of Gene Therapy Medicinal Products [2008]
FDA CBER Guidance for industry, “Considerations for Developmental Toxicity Studies 
for Preventive and Therapeutic Vaccines for Infectious Disease Indications. U.S. FDA; 
Feb. 2006. 
FDA Medical Devices Safety Alerts and Notices, “Blood human chorionic gonadotropin 
(hCG) assays: What laboratorians should know about false-positive results”, 

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http://www.fda.gov/MedicalDevices/Safety/AlertsandNotices/TipsandArticlesonDeviceS
afety/ucm109390.htm, accessed 17 Nov 2014.
Hatcher RA, Trussell J, Nelson AL, Cates W Jr, Stewart F, Kowal D, Policar MS, 
editors. Contraceptive Technology. 20th edition. Atlanta, Georgia: Ardent Media, Inc., 
2011: 50. Table 3-2.
International Conference on Harmonization of Technical Requirements for Registration 
of Pharmaceuticals For Human Use. ICH Harmonized Tripartite Guideline. Detection of 
Toxicity to Reproduction for Medicinal Products & Toxicity to Male Fertility. Parent 
Guideline dated 24 June 1993 (Addendum dated 9 November 2000 incorporated in 
November 2005. ICH S5 (R2)
International Conference on Harmonization of Technical Requirements for Registration 
of Pharmaceuticals For Human Use.  ICH Harmonized Tripartite Guideline. Nonclinical 
Evaluation for Anticancer Pharmaceuticals. ICH S9. 2009.
International Conference on Harmonization of Technical Requirements for Registration 
of Pharmaceuticals For Human Use. ICH Harmonized Tripartite Guideline. Nonclinical 
Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for 
Pharmaceuticals. ICH M3 (R2). 2009.
International Conference on Harmonization of Technical Requirements for Registration 
of Pharmaceuticals For Human Use. ICH Harmonized Tripartite Guideline. Preclinical 
Safety Evaluation of Biotechnology-Derived Pharmaceuticals. ICH S6 and Addendum 
ICH S6 (R1). 2011.
Kronenberg HM, Melmed S, Polonsky KS, Larsen PR, editors. Williams Textbook of 
Endocrinology, 11th edition. Philadelphia: Saunders, 2008.
Strauss JF, Barbieri RL, editors. Yen and Jaffe’s Reproductive Endocrinology. 5th 
edition, Philadelphia, Elsevier/Saunders, 2004.
U.S. Dept of Health and Human Services, FDA, Center For Biologics Evaluation and 
Research: Guidance for Human Somatic Cell Therapy and Gene Therapy [1998]
WHO, 2013 - WHO - Guidelines on the Nonclinical Evaluation of Vaccine Adjuvants 
and Adjuvanted Vaccines, 2013
World Health Organization. WHO/CONRAD Technical Consultation on Nonoxynol-9.
World Health Organization; Department of Reproductive Health and Research; Geneva; 
9-10 October 2001. Summary Report. 2001. WHO/RHR/03.8.

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12.6.
Appendix 6 - Country Specific Requirements
No country-specific requirements exist.

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12.7.
Appendix 7: Protocol Amendment Changes
AMENDMENT [4]
Where the Amendment Applies
This amendment applies across any site that will be used in the study.
Summary of Amendment Changes with Rationale
Section 4.4 Design Justification, has been updated to permit earlier interim analysis.
Section 5.3, Screening Failures/Re-Screening, has been updated to allow re-screening of 
Subjects screen failed.
Section 7.1.1 Time and Events Table – Screening Visit(s), has been updated to clarify 
timing of screening visits.
Section 10.2 Regulatory and Ethical Considerations, Including the Informed Consent 
Process, to clarify that a new ICF should be completed for each re-screened subject.
List of Specific Changes

Change to text in Section 4.4 Design Justification from Protocol Amendment 3 (26-
May-2017).
PREVIOUS TEXT
4.4
Design Justification

Interim analysis: Previous experience within GSK suggests some individuals 
(approximately 20%) fail to demonstrate a LAR response after the screening visit (i.e. 
they pass the screening criteria but do not demonstrate another LAR response when 
receiving placebo during the on-treatment phase). Those GSK studies were mainly 
crossover designs with a placebo in every sequence, so subjects who did not 
demonstrate a post screening LAR could be retrospectively identified and excluded 
from the statistical analyses. However this study has parallel groups and the profiles 
of a “perfect” drug effect and an inconsistent LAR subject are indistinguishable so 
retrospective exclusions cannot be made for the GSK2245035 arm. If the rate of 
“non-reproducible LAR” subjects is large it will reduce the probability of success. 
The purpose of the interim analysis is to evaluate whether the non-reproducible LAR 
rate is reducing the expected probability of success by more than 10% and, if it is, to 
perform an unblinded review of the observed data to assess the impact and cease 
further recruitment into the study, or enrol additional subjects to counteract the 
effects. Simulations were used to quantify the expected reduction in probability of 
success, using an estimate of the non-responder rate from the placebo arm only. The 
timing of the interim is linked to the enrolment rates and will be triggered by 
whichever occurs first i) Approximately 1 year after the first subject successfully 

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completes the Follow Up 1 assessments or, ii) When 28 subjects have successfully 
completed the Follow Up 1 assessments.
REVISED TEXT
4.4
Design Justification

Interim analysis: Previous experience within GSK suggests some individuals 
(approximately 20%) fail to demonstrate a LAR response after the screening visit (i.e. 
they pass the screening criteria but do not demonstrate another LAR response when 
receiving placebo during the on-treatment phase). Those GSK studies were mainly 
crossover designs with a placebo in every sequence, so subjects who did not 
demonstrate a post screening LAR could be retrospectively identified and excluded 
from the statistical analyses. However this study has parallel groups and the profiles 
of a “perfect” drug effect and an inconsistent LAR subject are indistinguishable so 
retrospective exclusions cannot be made for the GSK2245035 arm. If the rate of 
“non-reproducible LAR” subjects is large it will reduce the probability of success.
The purpose of the interim analysis is to evaluate whether the non-reproducible LAR 
rate is reducing the expected probability of success by more than 10% and, if it is, to 
perform an unblinded review of the observed data to assess the impact and cease 
further recruitment into the study, or enrol additional subjects to counteract the 
effects. Simulations were used to quantify the expected reduction in probability of 
success, using an estimate of the non-responder rate from the placebo arm only. The 
timing of the interim is linked to the enrolment rates and will be triggered by 
whichever occurs first i) Approximately 1 year after the first subject successfully 
completes the Follow Up 1 assessments or, ii) When 28 subjects have successfully 
completed the Follow Up 1 assessments or, iii) At the discretion of the GSK Study 
Team up to the time that 28 subjects have successfully completed the Follow Up 1 
assessment.

Change to text in Section 5.3, Screening Failures/Re-Screening from Protocol 
Amendment 3 (26-May-2017).
PREVIOUS TEXT
5.3
Screening Failures/Re-screening
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT) 
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.6).
Re-screening will not be permitted in this study. Sites are encouraged to screen subjects 
who have previously undergone BAC and have demonstrated a LAR. 

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REVISED TEXT
5.3
Screening Failures/Re-screening
Screen failures are defined as subjects who consent to participate in the clinical trial but 
are never subsequently randomized. In order to ensure transparent reporting of screen 
failure subjects, meet the Consolidated Standards of Reporting Trials (CONSORT) 
publishing requirements, and respond to queries from Regulatory authorities, a minimal 
set of screen failure information is required including Demography, Screen Failure 
details, Eligibility Criteria, and Serious Adverse Events (see Section 7.4.1.6).
Re-screening will be permitted in this study, in discussion with the Medical Monitor. 
Sites are encouraged to screen subjects who have previously undergone BAC and have 
demonstrated a LAR. 
Re-screened subjects should be assigned a separate number from their initial screening 
number and complete a new informed consent form. 

Section 7.1.1 Time and Events Table – Screening Visit(s), has been updated to clarify 
timing of screening visits. Protocol Amendment 3 (26-May-2017).
PREVIOUS TEXT
7.1.1 Time and Events Table – Screening Visit(s)
Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Informed consent signing
X
Includes genetics consent
Inclusion and exclusion 
criteria/eligibility review
X
X
Demographics
X
Medical history (including substance 
usage and family history of premature 
CV disease)
X
Past and current medical conditions 
including cardiovascular medical 
history
X
Concomitant medication review
X
Full physical exam including height 
and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all 
females)
X
No women of childbearing 
potential are being enrolled. 
However this test is needed to 
confirm pregnancy status at 
screening.
Serum FSH and estradiol for post-
menopausal females
X
HIV, Hep B and Hep C screen
X

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Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Laboratory assessments (including 
liver chemistries, haematology, clinical 
chemistry and urinalysis)
X
Screening Spirometry assessment
X
To confirm FEV1 >70% predicted
SAE Collection
X
X
X
12-lead ECG
X
Vital signs
X
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum 
induction
Saline and incremental BAC challenge 
plus associated spirometry 
measurements
X
Including pre-procedure FEV1
assessment and repeat FEV1
measurements up to 10 hours 
post BAC
Sputum induction and processing for 
BAC associated sample collection
X
X
Sputum inductions must be 
separated by at least 3 days
Laboratory assessments 
(haematology only)
X
X
Blood samples should be taken 
pre-BAC on SV2 and  on the 
following day at 24 hrs post BAC
Blood samples for PBMC preparations
X
Blood samples for PBMC 
preparations (pre-BAC and 6 
hours after BAC at screening visit 
2)
Provide diary card, PEF meters, 
rescue medication and thermometers 
to subjects
X
Only given out to subjects who 
are eligible following the BAC. 
Thermometers are to be given to 
record temperature as part of 
capturing AE information.
Nasal lavage
X
Collect nasal lavage to provide a 
background  sample. Repeat 
nasal lavage pre-NAC, then 
again 5 minutes and at least 6 
hours post-NAC.
Nasal filter collection 
X
Collect nasal filter sample to 
provide a background pre-wash 
sample, after nasal wash repeat 
nasal filter sampling pre-NAC, 
then again 5 minutes and at least 
6 hours post-NAC.
NAC
X
Commence after the 24-hour 
BAC sputum and blood 
biomarker samples have been 
taken.
Only to be done once subjects 
have completed a successful 
BAC.
ID challenge
X
ID challenge to be administered 
after NAC.
Measurements to be made 15 
minutes and 6 hours post ID 
challenge. 

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Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Nasal scrape
X
Nasal mucosa sample to be 
collected at least 6 hours post-
NAC.
CURRENT TEXT
7.1.1 Time and Events Table – Screening Visit(s)
Procedure
Screenin
g Visit 1 
(up to 
Day -28)
Screening 
Visit 2 (up to 
Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 to 
Day -2)
Notes 
There must be a minimum of 48 
hours between ‘Screening Visit 
1’ and ‘Screening Visit 2’. This 
ensures adherence to a 
minimum 3-day window between 
sputum inductions.
There must be a minimum of 48 
hours between ‘24 hours after 
Screening Visit 2’ and ‘Dosing 
Visit 1’.
Informed consent signing
X
Includes genetics consent
Inclusion and exclusion 
criteria/eligibility review
X
X
Demographics
X
Medical history (including substance 
usage and family history of premature 
CV disease)
X
Past and current medical conditions 
including cardiovascular medical 
history
X
Concomitant medication review
X
Full physical exam including height 
and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all 
females)
X
No women of childbearing 
potential are being enrolled. 
However this test is needed to 
confirm pregnancy status at 
screening.
Serum FSH and estradiol for post-
menopausal females
X
HIV, Hep B and Hep C screen
X
Laboratory assessments (including 
liver chemistries, haematology, clinical 
chemistry and urinalysis)
X
Screening Spirometry assessment
X
To confirm FEV1 >70% 
predicted
SAE Collection
X
X
X
12-lead ECG
X
Vital signs
X

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Procedure
Screenin
g Visit 1 
(up to 
Day -28)
Screening 
Visit 2 (up to 
Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 to 
Day -2)
Notes 
There must be a minimum of 48 
hours between ‘Screening Visit 
1’ and ‘Screening Visit 2’. This 
ensures adherence to a 
minimum 3-day window between 
sputum inductions.
There must be a minimum of 48 
hours between ‘24 hours after 
Screening Visit 2’ and ‘Dosing 
Visit 1’.
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum 
induction
Saline and incremental BAC challenge 
plus associated spirometry 
measurements
X
Including pre-procedure FEV1
assessment and repeat FEV1
measurements up to 10 hours 
post BAC
Sputum induction and processing for 
BAC associated sample collection
X
X
Sputum inductions must be 
separated by at least 3 days
Laboratory assessments 
(haematology only)
X
X
Blood samples should be taken 
pre-BAC on SV2 and  on the 
following day at 24 hrs post BAC
Blood samples for PBMC preparations
X
Blood samples for PBMC 
preparations (pre-BAC and 6 
hours after BAC at screening 
visit 2)
Provide diary card, PEF meters, 
rescue medication and thermometers 
to subjects
X
Only given out to subjects who 
are eligible following the BAC. 
Thermometers are to be given to 
record temperature as part of 
capturing AE information.
Nasal lavage
X
Collect nasal lavage to provide a 
background  sample. Repeat 
nasal lavage pre-NAC, then 
again 5 minutes and at least 6 
hours post-NAC.
Nasal filter collection 
X
Collect nasal filter sample to 
provide a background pre-wash 
sample, after nasal wash repeat 
nasal filter sampling pre-NAC, 
then again 5 minutes and at 
least 6 hours post-NAC.
NAC
X
Commence after the 24-hour 
BAC sputum and blood 
biomarker samples have been 
taken.
Only to be done once subjects 
have completed a successful 
BAC.
ID challenge
X
ID challenge to be administered 
after NAC.
Measurements to be made 15 
minutes and 6 hours post ID 
challenge. 
Nasal scrape
X
Nasal mucosa sample to be 
collected at least 6 hours post-

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Procedure
Screenin
g Visit 1 
(up to 
Day -28)
Screening 
Visit 2 (up to 
Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 to 
Day -2)
Notes 
There must be a minimum of 48 
hours between ‘Screening Visit 
1’ and ‘Screening Visit 2’. This 
ensures adherence to a 
minimum 3-day window between 
sputum inductions.
There must be a minimum of 48 
hours between ‘24 hours after 
Screening Visit 2’ and ‘Dosing 
Visit 1’.
NAC.

Changes to 10.2 Regulatory and Ethical Considerations, Including the Informed 
Consent Process from Protocol Amendment 3 (26-May-2017).
PREVIOUS TEXT
10.2
Regulatory and Ethical Considerations, Including the Informed 
Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.
The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent
CURRENT TEXT
10.2
Regulatory and Ethical Considerations, Including the Informed 
Consent Process
Prior to initiation of a site, GSK will obtain favourable opinion/approval from the 
appropriate regulatory agency to conduct the study in accordance with ICH Good Clinical 
Practice (GCP) and applicable country-specific regulatory requirements.
The study will be conducted in accordance with all applicable regulatory requirements, 
and with GSK policy.

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The study will also be conducted in accordance with ICH Good Clinical Practice (GCP), 
all applicable subject privacy requirements, and the guiding principles of the current 
version of the Declaration of Helsinki.  This includes, but is not limited to, the following:

IRB/IEC review and favorable opinion/approval of the study protocol and 
amendments as applicable

Obtaining signed informed consent (Subjects who are re-screened are required to 
sign a new ICF)

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Amendment [3]
Where the Amendment Applies
This amendment applies across any site that will be used in the study.
Summary of Amendment Changes with Rationale
Administrative changes including:

Changes in the Primary Medical Monitor and contact details.

Alignment of contraception requirements, throughout the document, for partners 
of male subjects that are female of reproductive potential.

Inclusion Criteria bullets replaced with numbers. 
List of Specific Changes
Changes in the Primary Medical Monitor and contact details.
PREVIOUS TEXT
MEDICAL MONITOR/SPONSOR INFORMATION PAGE
Medical Monitor/SAE Contact Information: 
Role
Name
Day Time Phone Number 
and email address
After-hours 
Phone/Cell/
Pager 
Number
Fax 
Number
Site Address
Primary 
Medical 
Monitor 
 
MD
5 Moore 
Drive, PO 
Box 13398, 
RTP, NC 
27709-3398, 
United 
States
Secondary 
Medical 
Monitor 
 
 MD
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
PPD
PPD
PPD
PPD
PPD

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Role
Name
Day Time Phone Number 
and email address
After-hours 
Phone/Cell/
Pager 
Number
Fax 
Number
Site Address
SAE contact  
information 
Medical 
monitor 
as above 
and 
Study 
Manager, 
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
REVISED TEXT
MEDICAL MONITOR/SPONSOR INFORMATION PAGE
Medical Monitor/SAE Contact Information: 
Role
Name
Day Time Phone Number 
and email address
After-hours 
Phone/Cell/
Pager 
Number
Fax 
Number
Site Address
Primary 
Medical 
Monitor 
 
 
MD
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
Secondary 
Medical 
Monitor 
 
 MD
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
SAE contact  
information 
Medical 
monitor 
as above 
and 
Study 
Manager, 
Gunnels 
Wood Road
Stevenage, 
SG1 2NY
UK
Section 5.1 Inclusion Criteria is now numbered. 
PPD
PPD
PPD
PPD
PPD
PPD
PPD
PPD

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PREVIOUS and REVISED TEXT
5.1 Inclusion Criteria. Bullets replaced with numbers 1-11.
Alignment of contraception requirements, throughout the document, for partners of male 
subjects that are female of reproductive potential.
PREVIOUS TEXT
5.1 Inclusion Criteria
SEX

Male OR female of non-reproductive potential
Males:
Male subjects with female partners of child bearing potential must comply with one of 
the following contraception requirements from the time of first dose of study medication 
until the final follow-up visit.
a.
Vasectomy with documentation of azoospermia. 
b. Male condom plus partner use of one of the contraceptive options below:

Contraceptive subdermal implant 

Intrauterine device or intrauterine system 

Oral contraceptive, either combined or progestogen alone [Hatcher, 2007a] 
Injectable progestogen [Hatcher, 2007a]

Contraceptive vaginal ring [Hatcher, 2007a]

Percutaneous contraceptive patches [Hatcher, 2007a]
This is an all-inclusive list of those methods that meet the following GSK definition of 
highly effective: having a failure rate of less than 1% per year when used consistently and 
correctly and, when applicable, in accordance with the product label. For non-product 
methods (e.g., male sterility), the investigator determines what is consistent and correct 
use. The GSK definition is based on the definition provided by the International 
Conference on Harmonization of Technical Requirements for Registration of 
Pharmaceuticals for Human Use (ICH) [ICH M3 (R2) 2009].”
The investigator is responsible for ensuring that subjects understand how to properly use 
these methods of contraception.
Females: 
A female subject is eligible to participate if she is not pregnant (as confirmed by a 
negative [serum or urine] human chorionic gonadotrophin (hCG) test), not lactating, and 
where the following condition applies:
Non-reproductive potential defined as:

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Pre-menopausal females with one of the following:
Documented tubal ligation
Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion
Hysterectomy
Documented Bilateral Oophorectomy
Postmenopausal defined as 12 months of spontaneous amenorrhea [in 
questionable cases a blood sample with simultaneous follicle stimulating 
hormone (FSH) and estradiol levels consistent with menopause (refer to 
laboratory reference ranges for confirmatory levels)].  Females on hormone 
replacement therapy (HRT) and whose menopausal status is in doubt will be 
required to use one of the highly effective contraception methods if they wish to 
continue their HRT during the study.  Otherwise, they must discontinue HRT to 
allow confirmation of post-menopausal status prior to study enrolment..
REVISED TEXT
SEX
10. Male OR female of non-reproductive potential
Males:
Male subjects with female partners of child bearing potential must comply with the 
contraception requirements in Appendix 5 (Modified List of Highly Effective Methods 
for Avoiding Pregnancy) from the time of first dose of study medication until the final 
follow-up visit.
This is an all-inclusive list of those methods that meet the following GSK definition of 
highly effective: having a failure rate of less than 1% per year when used consistently and 
correctly and, when applicable, in accordance with the product label. For non-product 
methods (e.g., male sterility), the investigator determines what is consistent and correct 
use. The GSK definition is based on the definition provided by the International 
Conference on Harmonization of Technical Requirements for Registration of 
Pharmaceuticals for Human Use (ICH) [ICH M3 (R2) 2009].”
The investigator is responsible for ensuring that subjects understand how to properly use 
these methods of contraception.
Females: 
A female subject is eligible to participate if she is not pregnant (as confirmed by a 
negative [serum or urine] human chorionic gonadotrophin (hCG) test), not lactating, and 
where the following condition applies:

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Non-reproductive potential defined as:
Pre-menopausal females with one of the following:
Documented tubal ligation
Documented hysteroscopic tubal occlusion procedure with follow-up 
confirmation of bilateral tubal occlusion
Hysterectomy
Documented Bilateral Oophorectomy
Postmenopausal defined as 12 months of spontaneous amenorrhea [in 
questionable cases a blood sample with simultaneous follicle stimulating 
hormone (FSH) and estradiol levels consistent with menopause (refer to 
laboratory reference ranges for confirmatory levels)].  Females on hormone 
replacement therapy (HRT) and whose menopausal status is in doubt will be 
required to use one of the highly effective contraception methods if they wish to 
continue their HRT during the study.  Otherwise, they must discontinue HRT to 
allow confirmation of post-menopausal status prior to study enrolment.
PREVIOUS TEXT
12.5.1       Modified List of Highly Effective Methods for Avoiding Pregnancy
Contraceptive requirements for male subjects with female partners of 
reproductive potential (when applicable).
Male subjects with female partners of child bearing potential must comply with the 
following contraception requirements from the time of first dose of study medication 
until [at least five half-lives of study medication OR for a cycle of spermatogenesis 
following five terminal half-lives] after the last dose of study medication.
REVISED TEXT
12.5.1       Modified List of Highly Effective Methods for Avoiding Pregnancy
Contraceptive requirements for male subjects with female partners of 
reproductive potential (when applicable).
Male subjects with female partners of child bearing potential must comply with the 
following contraception requirements from the time of first dose of study medication 
until the final follow-up visit.

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AMENDMENT [2]
Where the Amendment Applies
This amendment applies across any site that will be used in the study.
Summary of Amendment Changes with Rationale
The protocol title and text have been amended to clarify that while the investigator and 
subjects are blinded the sponsor-is unblinded. 
Details of the procedures required at early withdrawal visits have been added.
Details around the Safety Review Team have been added to include a consultant external 
to GSK.
The inclusion and exclusion criteria have been updated in Section 5.1 and Section 5.2. 
and clarification around prohibited medications has been provided in Section 6.11.2. to 
align with updated exclusion criteria (Section 5.2).
Administrative changes have been made to the T&E table (Section 7.1) and text to ensure 
consistency throughout.
List of Specific Changes
Title: Updated to reflect that the study is sponsor-unblind.
PREVIOUS TEXT
A randomised, double-blind placebo-controlled, parallel group, 8-week treatment study to 
investigate the safety, pharmacodynamics, and effect of the TLR7 agonist, GSK2245035, 
on the allergen-induced asthmatic response  in subjects with mild allergic asthma.
REVISED TEXT
A randomised, double-blind (sponsor open) placebo-controlled, parallel group, 8-week 
treatment study to investigate the safety, pharmacodynamics, and effect of the TLR7 
agonist, GSK2245035, on the allergen-induced asthmatic response  in subjects with mild 
allergic asthma.
This update has been made where necessary throughout the protocol.
Objectives/Endpoints: Updated to clarify what changes in FeNO measurement are 
being assessed; abbreviations for bronchial allergen challenge and nasal allergen 
challenge included. Nasal allergen administration changed to nasal allergen 
challenge for consistency.

To evaluate the induction of TLR7-
associated pharmacodynamic (PD) 

TLR7-induced blood PD biomarkers.

TLR7 induced gene expression changes 

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biomarkers and exhaled nitric oxide 
(FeNO) following treatment with i.n. 
GSK2245035
in blood

TLR7-induced nasal fluid PD biomarkers

FeNO levels 

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Bronchial Allergen Challenge 
associated allergic biomarkers

Allergic biomarkers in induced sputum

Allergic biomarkers in blood

FeNO levels

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Nasal Allergen Administration 
associated allergic biomarkers

Allergic biomarkers in nasal fluids

Cellular profile of nasal mucosa
REVISED TEXT

To evaluate the induction of TLR7-
associated pharmacodynamic (PD) 
biomarkers and exhaled nitric oxide 
(FeNO) following treatment with i.n. 
GSK2245035

TLR7-induced blood PD biomarkers.

TLR7 induced gene expression changes 
in blood

TLR7-induced nasal fluid PD biomarkers

Change in FeNO levels during the 
treatment phase

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Bronchial Allergen Challenge (BAC)
associated allergic biomarkers

Allergic biomarkers in induced sputum

Allergic biomarkers in blood

Change in FeNO levels following BAC

To evaluate the effect of treatment with 
i.n. GSK2245035 compared to placebo 
on Nasal Allergen Challenge(NAC)
associated allergic biomarkers

Allergic biomarkers in nasal fluids

Cellular profile of nasal mucosa
Bronchial Allergen Challenge has been replaced by BAC throughout the protocol and 
Nasal Allergen Challenge/Administration by NAC throughout the protocol. 
Section 4.3. Type and Number of Subjects, Final Paragraph updated
PREVIOUS TEXT
Accounting for replacements and interim analysis recommendations, the total number of 
subjects to be enrolled in this study is not expected to exceed 60

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REVISED TEXT
Accounting for replacements and interim analysis recommendations, the total number of 
subjects to be randomised in this study is not expected to exceed 60
Inclusion Criteria 5 updated to clarify timepoint of assessment
PREVIOUS TEXT
Pre-bronchodilator FEV1 > 70 % predicted normal.
REVISED TEXT
Pre-bronchodilator FEV1 > 70 % predicted normal at Screening Visit 1
Section 5.2. Exclusion Criteria
Two additional exclusion criteria have been added for clarification:

Use of long-acting antihistamines within 7 days’ or short-acting antihistamines 
within 72 hours prior to the screening skin prick test.

Patient known to be intolerant to salbutamol or albuterol
Section 5.4 Withdrawal/Stopping Criteria
PREVIOUS TEXT
Subjects withdrawn from the study while already exposed to IP will be asked to complete 
all safety assessments and the follow-up procedures. Relevant pages of the case report 
form (CRF) should be completed by the investigator.
REVISED TEXT
Subjects withdrawn from the study while already exposed to IP will be asked to complete 
all safety assessments as outlined in the Time and Events Table (Section 7.1.2). Relevant 
pages of the case report form (CRF) should be completed by the investigator.
Section 6.3 Blinding: An additional paragraph has been inserted between 
paragraphs 2 and 3.
PREVIOUS TEXT
This will be a double-blind study (investigators and subjects are blinded, and sponsor will 
be unblinded).
During the study, there will be an unblinded interim analysis. Some key GSK study team 
members may be unblinded as part of this analysis and review. Initially, only the study 
statistician(s) (and supporting programmers), project physician lead, and medical monitor 

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will be unblinded, and if further data reviews are required then details of who were un-
blinded to what data and when will be included in the clinical study report.
If the GSK medical monitor deems it necessary to unblind data in order to enhance the 
evaluation of safety and tolerability this will be permitted but also documented and 
discussed in the clinical study report.  The GSK medical monitor will also decide whether 
to un-blind the investigator(s)/site(s) at that time, or just the sponsor study staff.  Details 
of who were un-blinded to what data and when would then be included in the clinical 
study report.
REVISED TEXT
During the study, there will be an unblinded interim analysis. Some key GSK study team 
members may be unblinded as part of this analysis and review. Initially, only the study 
statistician(s) (and supporting programmers), project physician lead, and medical monitor 
will be unblinded, and if further data reviews are required then details of who were un-
blinded to what data and when will be included in the clinical study report.
On-going safety reviews will be conducted during the course of the study in an unblinded 
manner. Key study team members may be unblinded as part of this review. An external 
consultant, who is not a GSK employee, may also be unblinded to enable independent 
review of emerging safety data.
If the GSK medical monitor deems it necessary to unblind data in order to enhance the 
evaluation of safety and tolerability this will be permitted but also documented and 
discussed in the clinical study report.  The GSK medical monitor will also decide whether 
to un-blind the investigator(s)/site(s) at that time, or just the sponsor study staff.  Details 
of who were un-blinded to what data and when would then be included in the clinical 
study report.
Section 6.10.2. Caffeine, Alcohol, and Tobacco: A third bullet point has been added 
for clarification.
PREVIOUS TEXT

Subjects will abstain from ingesting excessive amounts (more than one cup) of 
caffeine- or xanthine-containing products (e.g. coffee, tea, cola drinks, chocolate) for 
6 hours prior to each dosing. 

During each visit in the clinic, subjects will abstain from alcohol for 24 hours prior 
to entry to the clinic until collection of all assessment is completed. 
REVISED TEXT

Subjects will abstain from ingesting excessive amounts (more than one cup) of 
caffeine- or xanthine-containing products (e.g. coffee, tea, cola drinks, chocolate) for 
6 hours prior to each dosing. 

During each visit in the clinic, subjects will abstain from alcohol for 24 hours prior 
to entry to the clinic until collection of all assessment is completed. 

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
Subjects will be reminded that all forms of smoking should be avoided during the 
course of the study
Section 6.11.2 Permitted Medications and Non-Drug Therapies: Additional wording
has been added to clarify which medications are permitted.
PREVIOUS TEXT
Any concomitant medications not specifically permitted will be considered on a case by 
case basis by the investigator and GSK medical monitor. 
Subjects are permitted to take inhaled short-acting β2-agonists (SABA) as rescue 
treatment, but must withhold them for at least 6 hours prior to study visits with BAC 
(screening and follow-up visits 1-3).
REVISED TEXT
Any concomitant medications not specifically permitted will be considered on a case by 
case basis by the investigator and GSK medical monitor. 
Subjects are permitted to take inhaled short-acting β2-agonists (SABA) as rescue 
treatment, but must withhold them for at least 6 hours prior to any study visits with lung 
function assessment or BAC (screening and follow-up visits 1-3).
Medication to control hypertension is permitted, providing it is anticipated to remain 
stable from screening to follow up. Beta blockers are not permitted.
Section 6.11.2 Prohibited Medications and Non-Drug Therapies. Clarification 
around the time period for which drugs are not permitted is provided.
PREVIOUS TEXT
Subjects may not receive the following treatments within 4 weeks prior to screening:

Intranasal steroids

Inhaled corticosteroid (ICS) with or without a long acting beta-2-agonist (LABA)

Non-ICS controller asthma medications (i.e. leukotriene modifier, 
methylxanthines)
Subjects are prohibited from taking antihistamines within 72 hours prior to the screening 
and follow-up bronchial allergen challenges.

Subjects are prohibited from taking systemic corticosteroids within 6 weeks prior to 
screening
REVISED TEXT
Subjects may not receive the following treatments from 4 weeks prior to screening until 
the final follow-up visit:

Intranasal steroids

Inhaled corticosteroid (ICS) with or without a long acting beta-2-agonist (LABA)

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
Non-ICS controller asthma medications (i.e. leukotriene modifier, 
methylxanthines).
Use of long-acting antihistamines within 7 days’ or short-acting antihistamines within 72 
hours prior to BAC, NAC or ID challenge is prohibited.
Subjects are prohibited from taking systemic corticosteroids within 6 weeks prior to 
screening.
Section 7.1 Time and Events Tables: These have been updated to provide 
consistency and clarification for the sites:
PREVIOUS TEXT
Section 7.1.1. Time and Events Table – Screening Visit(s)
Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Informed consent signing
X
Includes genetics consent
Inclusion and exclusion 
criteria/eligibility review
X
X
Demographics
X
Medical history (including substance 
usage and family history of premature 
CV disease)
X
Past and current medical conditions 
including cardiovascular medical 
history
X
Concomitant medication review
X
Full physical exam including height 
and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all 
females)
X
No women of childbearing 
potential are being enrolled. 
However this test is needed to 
confirm pregnancy status at 
screening.
Serum FSH and estradiol for post-
menopausal females
X
HIV, Hep B and Hep C screen
X
Laboratory assessments (including 
liver chemistries, haematology, clinical 
chemistry and urinalysis)
X
Screening Spirometry assessment
X
To confirm FEV1 > 70% predicted
12-lead ECG
X
Vital signs
X
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum 
induction
Saline and incremental BAC challenge 
plus associated spirometry 
X
Including baseline FEV1
assessment and repeat FEV1

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Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
measurements
measurements up to 10 hours 
post BAC
Sputum induction and processing for 
BAC associated sample collection
X
X
Sputum induction pre-BAC and 
24 hours after BAC: (must be 
separated by at least 3 days).
Blood sample for assessment of 
allergic biomarkers in blood
X1,2
X2
1 Collect blood samples for 
PBMC preparations (pre-BAC 
and 6 hours after BAC)
2 Collect blood sample for 
haematology (WBC count with 
differential) pre-BAC and 24 
hours after BAC
Provide diary card, PEF meters, 
rescue medication and thermometers 
to subjects
X
Only given out to subjects who 
are eligible following the BAC. 
Thermometers are to be given to 
record temperature as part of 
capturing AE information.
Nasal examination
X
NAC)
X
Commence after the 24-hour 
BAC sputum and blood 
biomarker samples have been 
taken ..
Only to be done once subjects 
have completed a successful 
BAC.
Collect sample for nasal baiomarker 
assessment
X
Collect samples pre-NAC, 5 
minutes and at 6 hours after NAC 
using nasal filters..
Nasal lavage
X
Collect nasal lavage pre-NAC, 5 
minutes and at least 6 hours after 
NAC.
Collect nasal mucosa sample at 
least 6 hours after NAC.
ID allergy testing
X
Intradermal challenge to be 
administered after NAC.
Measurements to be made 15 
minutes and 6 hours post 
intradermal challenge

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Section 7.1.2. Time and Events Table – Dosing visit (DV) 1 (Day 1) – DV8 (Week 8)
Procedure
Pre-
dose
Dos
e
Post-dose
Notes
0 
hour
20 
min
s
1 
ho
ur
12 
hours
24 
hours
Outpatient visit
X
X
Subjects get discharged 1-2 hours after 
dosing.
For DV1, 4, and 8, subjects will return to the 
unit 24 hours after dosing.
Inclusion and 
exclusion 
criteria/eligibility 
review
X
Only on DV1
12-lead ECG
X
X
X
Perform on DV1 and DV8 only
Vital signs (including 
temperature)
X1
X1
X2
1 Perform on DV1-8
2 Perform on DV1, 4 and DV8 only
Laboratory 
assessments 
(include liver 
chemistries, 
haematology, clinical 
chemistry and 
urinalysis)
X
X
Collect samples for laboratory assessments 
on DV1 and DV8
Blood sample for PK 
analysis
X
X
X
Collect samples on DV1, DV4 and DV8
Nasal examination
X
X
Conducted pre-dose and 24 hours post-dose 
on DV1 and DV8
Blood sample for 
biomarker analysis
X
X
Collect blood for serum samples and PBMC 
preparations pre-dose and 24 hrs post-dose 
on DV1 and DV8
Nasal lavage
X
X
Collect nasal lavage samples pre-dose and 
24hrs post-dose on DV1, DV4 and DV8
Blood PAX collection
X(3)
X(4)
PAX tubes for TLR7/AFFY chip gene 
expression in blood collected pre-dose on 
DV1(3) and 24 hrs after DV8(4)
Randomisation via 
IRT
X
Randomisation is on DV1 only
Study drug 
administration
X
All doses are administered in the unit.
Dispensing of study 
medication
X
One device will be assigned on DV1.  The 
second device will be assigned on DV5.  All 
devices remain in the unit.
AE/SAE review
X
Concomitant 
medication review
X
TNSS score (diary 
card) review
X
Captured daily AM and PM from DV1 to FUV2
PEF (diary card) 
review
X
Captured daily AM and PM from DV1 to FUV2
FeNO measurement
X
Perform FeNO measurements pre-dose on all 
dosing visits 1-8
Sputum induction 
and processing for 
BAC associated 
X
Sputum sample is conditional on having 
obtained a sputum sample from the subject at 
screening. Perform sputum induction after 

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Procedure
Pre-
dose
Dos
e
Post-dose
Notes
0 
hour
20 
min
s
1 
ho
ur
12 
hours
24 
hours
sample collection
collection of 24 hour post-dose DV8 blood 
and nasal samples (as pre BAC sample for 
FUV1).
Genetic sample
X
Take one sample on any of the Dosing Visits 
after genetics consent.
Diary card review
X
Review diary card at every dosing visit
Follow up phone call
X5
X6
5 Following at least the first two doses of study 
treatment to assess tolerability
6 On DV 2, 3, and 5-7, a follow-up phone call 
will be made approximately 24 hours post-
dose to check if any CRS events have 
occurred, and if they have, that they have 
been accurately recorded in the diary card 
and they will then be transcribed into the 
eCRF.
For DV1, 4 and 8, subjects will be in the unit 
24 hours after dosing, and any CRS events 
can be assessed and recorded directly by the 
site staff.
Section 7.1.3. Time and Events Table – Follow up visits (FUV1 to 3) (Weeks 9 to 20)
Procedure
Follow up 
Visit 1 
(FUV1)
Follow-up Visit 2 
(FUV2)
2 
mont
h 
follo
w-up 
phon
e call
(Wee
k 16)
Follow up Visit 3 
(FUV3)
Wee
k 9
(1 
wee
k 
afte
r 
DV8
)
24 
hou
rs 
late
r
Week 
11
(3 
week
s 
after 
DV8)
Week 
12
(4 
week
s 
after 
DV8)
24 
hou
rs 
late
r
Week 
19 
(11 
week
s 
after 
DV8)
Week 
20 
(12 
week
s 
after 
DV8)
24 
hour
s 
later
Notes
AE/SAE 
review
==========================
===============
X3
================
========
Concomitant 
medication 
review
==========================
===============
X3
================
========
3 Phone call made to 
the subject to check 
AEs and concomitant 
medications and 
confirm availability for 
FUV3
TNSS scores 
(diary card)
==========================
===============
Captured daily AM and 
PM up until the end of 
FUV 2

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Procedure
Follow up 
Visit 1 
(FUV1)
Follow-up Visit 2 
(FUV2)
2 
mont
h 
follo
w-up 
phon
e call
(Wee
k 16)
Follow up Visit 3 
(FUV3)
Wee
k 9
(1 
wee
k 
afte
r 
DV8
)
24 
hou
rs 
late
r
Week 
11
(3 
week
s 
after 
DV8)
Week 
12
(4 
week
s 
after 
DV8)
24 
hou
rs 
late
r
Week 
19 
(11 
week
s 
after 
DV8)
Week 
20 
(12 
week
s 
after 
DV8)
24 
hour
s 
later
Notes
PEF (diary 
card)
==========================
===============
X
Captured daily AM and 
PM up until the end of 
FUV2, and then 
checked again in the 
unit following the final 
BAC at FUV3
Diary card 
review
X
Review diary card at 
every visit until the end 
of FUV 2.
Blood PAX 
collection
X
PAX tubes for /AFFY 
chip gene expression in 
blood. Collected prior to 
the bronchial allergen 
challenge at FUV2
Saline and 
bolus BAC 
challenge plus 
associated 
spirometry 
measurement
s
X
X
X
Including baseline FEV1
assessment and repeat 
FEV1 measurements up 
to 10 hours post BAC
FeNO 
measurement
X
X
X
X
X
X
Collect prior to sputum 
induction
Sputum 
induction and 
processing for 
BAC 
associated 
sample 
collection of 
allergic 
biomarkers
X
X
X
X
X
Sputum induction pre-
BAC and 24 hours after 
BAC.
BAC 
associated 
sample 
collection of 
allergic 
biomarkers in 
blood
X1,2
X2
X1,2
X2
X1,2
X2
1Blood samples for 
PBMC preparations 
(pre-BAC and 6 hours 
after BAC)
2 Blood sample for 
haematology (WBC 
count with differential) 
pre-BAC and 24 hours 
after BAC

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Procedure
Follow up 
Visit 1 
(FUV1)
Follow-up Visit 2 
(FUV2)
2 
mont
h 
follo
w-up 
phon
e call
(Wee
k 16)
Follow up Visit 3 
(FUV3)
Wee
k 9
(1 
wee
k 
afte
r 
DV8
)
24 
hou
rs 
late
r
Week 
11
(3 
week
s 
after 
DV8)
Week 
12
(4 
week
s 
after 
DV8)
24 
hou
rs 
late
r
Week 
19 
(11 
week
s 
after 
DV8)
Week 
20 
(12 
week
s 
after 
DV8)
24 
hour
s 
later
Notes
Nasal allergen 
challenge/adm
inistration
X
X
Commence after the 
24-hour BAC sputum 
and blood biomarker  
samples have been 
taken at FUV1 and 
FUV2
Nasal filter 
biomarker 
collection
X
X
Collect pre-NAC, 5 
minutes and 6 hours 
after NAC.
Nasal fluid 
biomarker 
collection
X
X
Collect nasal lavage 
pre-NAC, 5 minutes and 
at least 6 hours after 
NAC.
Collect nasal mucosa 
sample at least 6 hours 
after NAC.
Intradermal 
allergy testing
X
X
Intradermal challenge to 
be administered after 
NAC.
Measurements to be 
done at 15 minutes and 
6 hours post 
intradermal challenge.
Return diary 
card, PEF 
meters, 
rescue 
medication 
and 
thermometers
X

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REVISED TEXT
Section 7.1.1. Time and Events Table – Screening Visit(s)
Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Informed consent signing
X
Inclusion and exclusion 
criteria/eligibility review
X
X
Demographics
X
Medical history (including substance 
usage and family history of premature 
CV disease)
X
Past and current medical conditions 
including cardiovascular medical 
history
X
Concomitant medication review
X
Full physical exam including height 
and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all 
females)
X
No women of childbearing 
potential are being enrolled. 
However this test is needed to 
confirm pregnancy status at 
screening.
Serum FSH and estradiol for post-
menopausal females
X
HIV, Hep B and Hep C screen
X
Laboratory assessments (including 
liver chemistries, haematology, clinical 
chemistry and urinalysis)
X
Screening Spirometry assessment
X
To confirm FEV1 > 70% predicted
SAE Collection
X
X
X
12-lead ECG
X
Vital signs
X
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum 
induction
Saline and incremental BAC challenge 
plus associated spirometry 
measurements
X
Including pre-procedure FEV1
assessment and repeat FEV1
measurements up to 10 hours 
post BAC
Sputum induction and processing for 
BAC associated sample collection
X
X
Sputum inductions must be 
separated by at least 3 days
Laboratory assessments 
(haematology only)
X
X
Blood samples should be taken 
pre-BAC on SV2 and  on the 
following day at 24hrs post BAC
Blood samples for PBMC preparations
X
Blood samples for PBMC 
preparations (pre-BAC and 6 
hours after BAC at screening visit 
2)

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Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Provide diary card, PEF meters, 
rescue medication and thermometers 
to subjects
X
Only given out to subjects who 
are eligible following the BAC. 
Thermometers are to be given to 
record temperature as part of 
capturing AE information.
NAC
X
Commence after the 24-hour 
BAC sputum and blood 
biomarker samples have been 
taken.
Only to be done once subjects 
have completed a successful 
BAC.
Nasal lavage
X
Collect nasal lavage to provide a 
background  sample. Repeat 
nasal lavage pre-NAC, then 
again 5 minutes and at least 6 
hours post-NAC.
Nasal filter collection 
X
Collect nasal filter sample to 
provide a background pre-wash 
sample, after nasal wash repeat 
nasal filter sampling pre-NAC, 
then again 5 minutes and at least 
6 hours post-NAC.
Nasal scrape
X
Nasal mucosa sample to be 
collected at least 6 hours post-
NAC.
ID challenge
X
ID challenge to be administered 
after NAC.
Measurements to be made 15 
minutes and 6 hours post ID 
challenge

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Section 7.1.2. Time and Events Table – Dosing visit (DV) 1 (Day 1) – DV8 (Week 8) 
and Early Withdrawal Visit (EW)
Procedure
Pre-
dose
Dose
Post-dose
Early-
Withd
rawal
Notes
0 
hour
20 
mins
1 hour
12 
hours
24 
hours
Outpatient 
visit
X
X
X
Subjects get discharged 1-2 
hours after dosing.
Inclusion and 
exclusion 
criteria/eligibil
ity review
X
Only on DV1
Diary card 
collection and 
review 
X
X
Collect and review diary card 
at every dosing visit 
AE/SAE 
review
X
X
Concomitant 
medication 
review
X
X
12-lead ECG
X
X
X
X
Perform on DV1 and DV8 
only
Vital signs 
(including 
temperature)
X1
X1
X2
X
1 Perform on DV1-8
2 Perform on DV1, 4 and DV8 
only
Laboratory 
assessments 
(include liver 
chemistries, 
haematology, 
clinical 
chemistry and 
urinalysis)
X
X
X
Collect samples for laboratory 
assessments on DV1 and 
DV8
Not required at EW if this 
occurs during FU
Blood sample 
for PK 
analysis
X
X
X
Collect samples on DV1, DV4 
and DV8.
Not required at EW if this 
occurs during FU
Genetic 
sample
X
Take one sample on any of 
the dosing visits after 
genetics consent.
Not required at EW if this 
occurs during FU
FeNO 
measurement
X
Perform FeNO 
measurements pre-dose on 
all dosing visits 1-8
Nasal 
examination
X
X
Conducted pre-dose at all 
study visits and 24 hours 
post-dose on DV1, DV4 and 
DV8
Nasal lavage
X
X
Collect nasal lavage samples 
pre-dose and 24hrs post-
dose on DV1, DV4 and DV8

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Procedure
Pre-
dose
Dose
Post-dose
Early-
Withd
rawal
Notes
0 
hour
20 
mins
1 hour
12 
hours
24 
hours
Blood sample 
for biomarker 
analysis
X
X
Collect blood for serum 
samples and PBMC 
preparations pre-dose and 24 
hrs post-dose on DV1 and 
DV8
Blood sample 
for RNA 
analysis 
X(3)
X(4)
PAX-gene tubes for 
TLR7/AFFY chip gene 
expression in blood collected 
pre-dose on DV1(3) and 24 
hrs after DV8(4)
Call RAMOS
X
Randomisation will occur is at 
Visit 1 via the IRT. Sites 
should also call RAMOS at 
DV5.
Dispensing of 
study 
medication
X
One device will be assigned 
on DV1. The second device 
will be assigned on DV5.  All 
devices remain in the unit.
Study drug 
administration
X
All doses are administered in 
the unit.
Sputum 
induction and 
processing 
for BAC 
associated 
sample 
collection
X
Sputum sample is conditional 
on having obtained a sputum 
sample from the subject at 
screening. Perform sputum 
induction after collection of 
24 hour post-dose DV8 blood 
and nasal samples (as pre 
BAC sample for FUV1).
Follow up 
phone call
X5
X6
5 Following at least the first 
two doses of study treatment 
to assess tolerability
6 On DV 2, 3, and 5-7, a 
follow-up phone call will be 
made approximately 24 hours 
post-dose to check if any 
CRS events have occurred, 
and if they have, that they 
have been accurately 
recorded in the diary card 
and they will then be 
transcribed into the eCRF.
For DV1, 4 and 8 the 
subjects shall be assessed 
on their return to the unit the 
following day.

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Section 7.1.3. Time and Events Table – Follow up visits (FUV1 to 3) (Weeks 9 to 20)
Procedure
Follow up 
Visit 1 
(FUV1)
Follow-up Visit 2 
(FUV2)
2 
mont
h 
follo
w-up 
phon
e call
(Wee
k 16)
Follow up Visit 3 
(FUV3)
Wee
k 9
(1 
wee
k 
afte
r 
DV8
)
24 
hou
rs 
late
r
Week 
11
(3 
week
s 
after 
DV8)
Week 
12
(4 
week
s 
after 
DV8)
24 
hou
rs 
late
r
Week 
19 
(11 
week
s 
after 
DV8)
Week 
20 
(12 
week
s 
after 
DV8)
24 
hour
s 
later
Notes
AE/SAE 
review
==========================
===============
X3
================
========
Concomitant 
medication 
review
==========================
===============
X3
================
========
3 Phone call made to 
the subject to check 
AEs and concomitant 
medications and 
confirm availability for 
FUV3
Diarycard 
collection, 
review and 
dispensing
X
Review diary card at 
every visit until the end 
of FUV 2
FeNO 
measurement
X
X
X
X
X
X
Collect prior to sputum 
induction
Blood sample 
for RNA 
analysis
X
PAX tubes for /AFFY 
chip gene expression in 
blood. Collected prior to 
the bronchial allergen 
challenge at FUV2
Saline and 
bolus BAC 
challenge plus 
associated 
spirometry 
measurement
s
X
X
X
Including pre-procedure 
FEV1 assessment and 
repeat FEV1
measurements up to 10 
hours post BAC
Sputum 
induction and 
processing for 
BAC 
associated 
sample 
collection
X
X
X
X
X
Sputum induction pre-
BAC and 24 hours after 
BAC
Laboratory 
assessments 
(haematology 
only)
X
X
X
X
X1
X
Blood samples should 
be taken pre-BAC  and  
on the following day at 
24hrs post BAC
Blood 
samples for 
PBMC 
preparations
X
X
X
Blood samples for 
PBMC preparations 
(pre-BAC and 6 hours 
after BAC)

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Procedure
Follow up 
Visit 1 
(FUV1)
Follow-up Visit 2 
(FUV2)
2 
mont
h 
follo
w-up 
phon
e call
(Wee
k 16)
Follow up Visit 3 
(FUV3)
Wee
k 9
(1 
wee
k 
afte
r 
DV8
)
24 
hou
rs 
late
r
Week 
11
(3 
week
s 
after 
DV8)
Week 
12
(4 
week
s 
after 
DV8)
24 
hou
rs 
late
r
Week 
19 
(11 
week
s 
after 
DV8)
Week 
20 
(12 
week
s 
after 
DV8)
24 
hour
s 
later
Notes
Nasal lavage
X
X
Collect nasal lavage to 
provide a background 
sample. Repeat nasal 
lavage pre-NAC, then 
again 5 minutes and at 
least 6 hours post-NAC.
Nasal filter 
collection
X
X
Collect nasal filter 
sample to provide a 
background pre-wash 
sample, after nasal 
wash repeat nasal filter 
sampling pre-NAC, then 
again 5 minutes and at 
least 6 hours post-NA.
NAC
X
X
Commence after the 
24-hour BAC sputum 
and blood biomarker  
samples have been 
taken at FUV1 and 
FUV2
ID challenge
X
X
ID challenge to be 
administered after NAC.
Measurements to be 
made at 15 minutes and 
6 hours post ID 
challenge.
Nasal scrape
X
X
Nasal mucosa sample 
to be collected at least 
6 hours post-NAC
Return diary 
card and 
thermometers
X
Return PEF 
meters and 
rescue 
medication
X
Section 7.3.1. Bronchial allergen challenges and spirometry: This section has been 
simplified and the detail moved to the SRM
PREVIOUS TEXT
Bronchial allergen challenges will be conducted at screening, and at three follow-up 
visits (see Time and Events Tables in Section 7.1).

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Baseline FEV1 measurements for the challenge will first be taken using saline. The 
highest of the measurements will be taken as the saline baseline value and recorded in the 
eCRF. This value will be the baseline for the allergen challenge. If a fall in FEV1 of 
>10% is observed after saline, the subject will not proceed to allergen inhalation. At 
screening, subjects will inhale increasing concentrations of allergen during the bronchial 
allergen challenge. FEV1 will be measured after each inhalation. The final concentration 
of allergen will be determined when a fall in FEV1 of ≥ 20 % from the saline baseline 
value is achieved. For the subject to be eligible, the screening bronchial allergen 
challenge must demonstrate both an early and late asthmatic response. The early 
asthmatic response must include a fall in FEV1 of 20% from the saline value, on at least 
one occasion, between 5 and 30 minutes after the final concentration of allergen. The late 
asthmatic response must include a fall in FEV1 of 15% from the saline value, on at least 
three occasions, two of which must be consecutive, between 4 and 10 hours after the final 
concentration of allergen. FEV1 measurements will be taken at specified time intervals 
after the final inhalation of allergen up to 10 hours.
Following the final FEV1 measurement, the subject will receive rescue salbutamol until 
the FEV1 returns to within 10% of the pre-saline value. If a subject experiences serious 
discomfort the test will be discontinued and rescue medication administered.
The allergen dose eliciting a fall in FEV1 of ≥ 20 % from the saline baseline value at 
screening will be used for bronchial allergen challenges administered at the follow-up 
visits and FEV1 measurements will be taken at specified time intervals up to 10 hours. 
Detailed description of the bronchial allergen challenge technique and assessments can be 
found in the SRM
REVISED TEXT
BAC will be conducted at screening, and at three follow-up visits (see Time and Events 
Tables in Section 7.1). At the Screening Visit incremental doses of allergen will be 
administered and the total dose inducing an EAR and LAR used to calculate a bolus dose 
for use at the 3 FU visits. A detailed description of the BAC technique and assessments 
can be found in the SRM.  All sites participating in the study will be trained in the same 
methodology to ensure consistency.
Following the final FEV1 measurement, subjects will receive rescue salbutamol until the 
FEV1 returns to within 10% of the pre-saline value. If a subject experiences serious 
discomfort the test will be discontinued and rescue medication administered.
Section 7.3.2. Intranasal and intradermal allergen challenge
PREVIOUS TEXT
Intranasal and intradermal allergen challenges will be conducted approximately 24 hours 
after the bronchial allergen challenge at screening and at follow-up visits 1 and 2. These 
challenges will only be conducted after subjects have completed successful bronchial 
allergen challenges.

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A detailed description of the intranasal and intradermal allergen challenges can be found 
in the SRM.
In some regions/countries specific challenges agents may not be approved or available 
for use in ID challenge, in which case this assessment shall be omitted.
REVISED TEXT
NAC and ID challenge will be conducted approximately 24 hours after the bronchial 
allergen challenge at screening and at follow-up visits 1 and 2. 
A detailed description of the NAC and ID challenges can be found in the SRM.
Section 7.3.4. TNSS: Updated as transcription to CRF does not necessarily have to 
be done by site staff.
PREVIOUS TEXT
All data will be transcribed from the diary card into the CRF by the site staff at each 
clinic visit
REVISED TEXT
All data will be transcribed from the diary booklet into the CRF.
Section 7.3.6. Fractional Exhaled Nitric Oxide (FeNO): Updated to reflect that 
equipment will not be provided centrally.
PREVIOUS TEXT
FeNO will be measured using a handheld electronic device.  Measurements will be 
obtained in accordance with the [ATS/ERS Recommendations for Standardized 
Procedures for the Online and Offline Measurement of Exhaled Lower Respiratory Nitric 
Oxide and Nasal Nitric Oxide, 2005].  All sites will use standardised equipment provided 
by a central vendor.  FeNO measurements will be interpreted in accordance with the 
[Official ATS Clinical Practice Guideline: Interpretation of Exhaled Nitric Oxide Levels 
(FeNO) for Clinical Applications, 2011].
REVISED TEXT
FeNO will be measured using a handheld electronic device.  Measurements will be 
obtained in accordance with the [ATS/ERS Recommendations for Standardized 
Procedures for the Online and Offline Measurement of Exhaled Lower Respiratory Nitric 
Oxide and Nasal Nitric Oxide, 2005]. FeNO measurements will be interpreted in 
accordance with the [Official ATS Clinical Practice Guideline: Interpretation of Exhaled 
Nitric Oxide Levels (FeNO) for Clinical Applications, 2011].

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Section 7.4. Safety: Updated to describe the role of the SRT and to reflect the 
inclusion of an external consultant.
PREVIOUS TEXT
Planned time points for all safety assessments are listed in the Time and Events Table 
(Section 7.1).  Additional time points for safety tests (such as vital signs, physical exams 
and laboratory safety tests) may be added during the course of the study based on newly 
available data to ensure appropriate safety monitoring
REVISED TEXT
Planned time points for all safety assessments are listed in the Time and Events Table 
(Section 7.1).  Additional time points for safety tests (such as vital signs, physical exams 
and laboratory safety tests) may be added during the course of the study based on newly 
available data to ensure appropriate safety monitoring.
The Safety Review Team (SRT) is a GSK cross-functional team reviewing all available 
safety data related to the project, including in-stream data from this study, in an ongoing 
manner. The SRT is an internal GSK requirement put in place to ensure holistic 
evaluation of the safety profile of an investigational product with systematic, periodic and 
documented reviews of available safety data, with the appropriate communication and 
escalation of new findings that have the potential to impact patient safety.  
The SRT for this project will review data from this study in collaboration with an 
independent external (to GSK) expert reviewer.
Section 7.4.7. PEF monitoring: Final Paragraph updated as transcription to CRF 
does not necessarily have to be done by site staff.
PREVIOUS TEXT
All information from the diary card will be transcribed into the eCRF by the site staff at 
each clinic visit
REVISED TEXT
All information from the diary card will be transcribed into the eCRF.
Section 7.4.7.: PEF monitoring: Text updated to clarify that these are being used for 
safety monitoring at home. Assessment in clinic at FV3 will not be required as 
subject will be attending the clinic the day after their final BAC. 
PREVIOUS TEXT
Subjects will be given a PEF peak flow meter at screening, along with a diary card. 
Subjects will record their PEF readings each morning and evening onto the diary card 
every day from the start of study treatment on Dosing Visit 1 until the 4 week follow-up
visit. Subjects will also have their PEF levels checked in the unit after administration of 
the final bronchial allergen challenge at the 12 week follow-up visit.

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REVISED TEXT
Subjects will be given a PEF peak flow meter at screening, along with a diary card. This 
will be used for safety monitoring after the BACS. Subjects will record their PEF 
readings each morning and evening onto the diary card every day from the start of study 
treatment on DV 1 until the 4 week follow-up visit.  
The subject will be instructed to contact the investigator if at any time during the study 
the PEF measurements go outside the normal range for the subject. These instructions 
will be clearly printed on the diary card.
Section 7.6.1.1. PD blood samples
PREVIOUS TEXT
Blood samples will be collected before and 24 hours post treatment on Dosing Visits 1 
and 8 for serum to measure levels of TLR-7-induced blood PD biomarkers. 
Blood will be collected before and 24 hours after treatment on Dosing Visits 1 and 8 and 
prior to bronchial allergen challenges for preparation of peripheral blood mononuclear 
cells (PBMC). These samples may be used for CHIP cytometry for cellular subset 
phenotyping. If this biomarker analysis is performed it may be reported separately.
REVISED TEXT
Blood samples will be collected before and 24 hours post treatment on DV 1 and 8 for 
serum to measure blood PD biomarkers. 
Blood will be collected before and 24 hours after treatment on DV 1 and 8 and prior to 
bronchial allergen challenges for preparation of peripheral blood mononuclear cells 
(PBMC). These samples may be used for CHIP cytometry for cellular subset 
phenotyping. If this biomarker analysis is performed it may be reported separately.
Section 7.6.1.2. Blood PAX collection
PREVIOUS TEXT
Blood PAX samples for AFFYCHIP gene expression (mRNA transcriptome) will be 
collected before dosing on Dosing Visit 1, 24 hours after Dosing Visit 8 and prior to 
BAC on FUV2.
REVISED TEXT
Blood PAX samples for AFFYCHIP gene expression (mRNA transcriptome) will be 
collected before dosing on DV 1, 24 hours after DV 8 and prior to BAC on FUV2.

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Section 7.6.1.3. PD nasal lavage samples
PREVIOUS TEXT
Nasal lavage samples to measure TLR-7-induced nasal fluid PD biomarkers will be 
collected before and 24 hours post treatment on Dosing Visits 1, 4 and 8.
REVISED TEXT
Nasal lavage samples to measure PD biomarkers will be collected before and 24 hours 
post treatment on DV1, DV4 and DV8.
Section 7.6.1.6. :Nasal allergen challenge associated biomarkers in nasal fluid 
samples
PREVIOUS TEXT
Nasal filter and nasal lavage samples to measure exploratory allergic biomarkers will be 
collected at baseline, then after a nasal wash, samples will be collected before the nasal 
allergen administration (NAC), and at 5 minutes and 6 hours after NAC (at screening and 
at FUV1 and FUV2).
REVISED TEXT
Nasal filter and nasal lavage samples to measure exploratory biomarkers will be collected 
at the timepoints indicated in the Time and Events Tables Section 7.1.
Section 7.6.1.7. Cellular profile of nasal mucosa
PREVIOUS TEXT
Nasal scrapes will be collected after NAC following the completion of the collection of 
the 6 hour nasal lavage and nasal filter samples (at screening and at FUV1 and FUV2). 
Samples of suitable quality may be used for cellular analysis by CHIP cytometry or 
RNA-sequencing of single cells (if subjects have given informed consent for genetic 
analysis techniques).  If this biomarker analysis is performed it may be reported 
separately.
REVISED TEXT
Nasal scrapes may be collected after NAC following the completion of the collection of 
the 6 hour nasal lavage and nasal filter samples (at screening and at FUV1 and FUV2). 
Samples of suitable quality may be used for cellular analysis for example by CHIP 
cytometry or RNA-sequencing of single cells (if subjects have given informed consent 
for genetic analysis techniques).  If this biomarker analysis is performed it may be 
reported separately.

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Section 7.7.1.5.:Bronchial allergen challenge (BAC) associated biomarkers in blood 
samples – Updated to reflect that RNA analysis of single cells will not be performed.
PREVIOUS TEXT
Blood samples will be collected before, 6 hours and 24 hours after each BAC:

For haematology, (WBC with differential) for blood eosinophil counts (before and 
24 hours after BAC)

To prepare PBMC that may be used for CHIP cytometry or RNA sequencing of 
single cells (if patients give informed consent for genetic analysis techniques) before 
and 6 hours after BAC.  If this biomarker analysis is performed it may be reported 
separately.
REVISED TEXT
Blood samples will be collected before, 6 hours and 24 hours after each BAC:

For haematology, (WBC with differential) for blood eosinophil counts (before and 
24 hours after BAC)

To prepare PBMC that may be used for CHIP cytometry (before and 6 hours after 
BAC).  If this biomarker analysis is performed it may be reported separately.

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AMENDMENT [1]
Where the Amendment Applies
This amendment applies across any site that will be used in the study.
Summary of Amendment Changes with Rationale
Section 4.6.1 and Section 7.1.1 have been adjusted slightly to make it clearer that subjects 
will be provided with a thermometer to be able to record temperature as part of capturing 
adverse event (AE) information.
In Section 5.2, exclusion criterion 13 has been updated to reflect the revised 
recommended alcohol consumption guidelines.
List of Specific Changes
Section 4.6.1, Risk assessment table, first paragraph
PREVIOUS TEXT
Each subject will receive a phone call approximately 12 hours following administration 
of at least the first two doses to assess tolerability. They will be provided with a 
thermometer to measure body temperature as needed.
REVISED TEXT
Each subject will receive a phone call approximately 12 hours following administration 
of at least the first two doses to assess tolerability. They will be provided with a 
thermometer to measure body temperature.
Section 5.2, Exclusion criteria, point 13
PREVIOUS TEXT
RELEVANT HABITS
13. History of regular alcohol consumption within 6 months of the study defined as: 
An average weekly intake of >21 units for males or >14 units for females.  One unit 
is equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 ml) of wine 
or 1 (25 ml) measure of spirits.
REVISED TEXT
RELEVANT HABITS
13. History of regular alcohol consumption within 6 months of the study defined as: 
An average weekly intake of >14 units for males and females.  One unit is 
equivalent to 8 g of alcohol: a half-pint (~240 ml) of beer, 1 glass (125 ml) of wine or 
1 (25 ml) measure of spirits.

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Section 7.1.1, Time and Events table, Screening Visit(s)
PREVIOUS TEXT
Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Informed consent signing
X
Inclusion and exclusion 
criteria/eligibility review
X
X
Demographics
X
Medical history (including substance 
usage and family history of premature 
CV disease)
X
Past and current medical conditions 
including cardiovascular medical 
history
X
Concomitant.medication review
X
Full physical exam including height 
and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all 
females)
X
No women of childbearing 
potential are being enrolled. 
However this test is needed to 
confirm pregnancy status at 
screening.
Serum FSH and estradiol for post-
menopausal females
X
HIV, Hep B and Hep C screen
X
Laboratory assessments (including 
liver chemistries, haematology, clinical 
chemistry and urinalysis)
X
Spirometry
X
To confirm FEV1 > 70% predicted
12-lead ECG
X
Vital signs
X
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum 
induction
BAC and spirometry
X
Including repeat FEV1  
measurements up to 10 hours 
post BAC
Sputum induction and processing for 
BAC associated sample collection
X
X
Sputum induction pre-BAC and 
24 hours after BAC
BAC associated sample collection of 
allergic biomarkers in blood
X1,2
X2
1 Collect blood samples for 
PBMC preparations (pre-BAC 
and 6 hours after BAC)
2 Collect blood sample for 
haematology (WBC count with 
differential) pre-BAC and 24 
hours after BAC
Provide diary card, PEF meters, 
rescue medication and thermometers 
to subjects
X
Only given out to subjects who 
are eligible following the BAC. 
Thermometers are to be given to 

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Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
record temperature as part of 
capturing AE information, if 
needed.
Nasal allergen challenge 
administration (NAC)
X
Commence after the 24-hour 
BAC sputum and blood 
biomarker  samples have been 
taken ..
Only to be done once subjects 
have completed a successful 
BAC.
Nasal filter biomarker collection
X
Collect pre-NAC, 5 minutes and 
at 6 hours after NAC.
Nasal fluid biomarker collection
X
Collect nasal lavage pre-NAC, 5 
minutes and at least 6 hours after 
NAC.
Collect nasal mucosa sample at 
least 6 hours after NAC.
Intradermal allergy testing
X
Intradermal challenge to be 
administered after NAC.
Measurements to be made 15 
minutes and 6 hours post 
intradermal challenge
REVISED TEXT
Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
Informed consent signing
X
Inclusion and exclusion 
criteria/eligibility review
X
X
Demographics
X
Medical history (including substance 
usage and family history of premature 
CV disease)
X
Past and current medical conditions 
including cardiovascular medical 
history
X
Concomitant.medication review
X
Full physical exam including height 
and weight
X
Urine drug/alcohol screen
X
Urine or serum pregnancy test (all 
females)
X
No women of childbearing 
potential are being enrolled. 
However this test is needed to 
confirm pregnancy status at 

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Procedure
Screenin
g Visit 1 
(Day -28 
to Day -
22)
Screening 
Visit 2 (Day -
21 to Day -3)
24 hours 
after 
Screening 
Visit 2 
(Day -20 
to Day -2)
Notes
screening.
Serum FSH and estradiol for post-
menopausal females
X
HIV, Hep B and Hep C screen
X
Laboratory assessments (including 
liver chemistries, haematology, clinical 
chemistry and urinalysis)
X
Spirometry
X
To confirm FEV1 > 70% predicted
12-lead ECG
X
Vital signs
X
Skin prick test
X
To confirm eligibility
FeNO measurement
X
X
Collected prior to sputum 
induction
BAC and spirometry
X
Including repeat FEV1  
measurements up to 10 hours 
post BAC
Sputum induction and processing for 
BAC associated sample collection
X
X
Sputum induction pre-BAC and 
24 hours after BAC
BAC associated sample collection of 
allergic biomarkers in blood
X1,2
X2
1 Collect blood samples for 
PBMC preparations (pre-BAC 
and 6 hours after BAC)
2 Collect blood sample for 
haematology (WBC count with 
differential) pre-BAC and 24 
hours after BAC
Provide diary card, PEF meters, 
rescue medication and thermometers 
to subjects
X
Only given out to subjects who 
are eligible following the BAC. 
Thermometers are to be given to 
record temperature as part of 
capturing AE information.
Nasal allergen challenge 
administration (NAC)
X
Commence after the 24-hour 
BAC sputum and blood 
biomarker samples have been 
taken ..
Only to be done once subjects 
have completed a successful 
BAC.
Nasal filter biomarker collection
X
Collect pre-NAC, 5 minutes and 
at 6 hours after NAC.
Nasal fluid biomarker collection
X
Collect nasal lavage pre-NAC, 5 
minutes and at least 6 hours after 
NAC.
Collect nasal mucosa sample at 
least 6 hours after NAC.
Intradermal allergy testing
X
Intradermal challenge to be 
administered after NAC.
Measurements to be made 15 
minutes and 6 hours post 
intradermal challenge