Clinical trial NCT02660489
Conditions: Asthma Rhinovirus Picornaviridae Infections Common Cold
Keywords: CRTH2 antagonist Prostaglandin D2 receptor Anti-Asthmatic Agents OC459 Allergic asthma Eosinophilia
Sponsor: Imperial College London
Start date: 2015-01-01; Completion date: 2018-02-01
Enrollment: 44 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The aim of this study is to assess the effectiveness of a CRTH2 receptor antagonist, OC459, in preventing or attenuating the worsening of asthma symptoms during rhinovirus infection. The study is a double blind, randomised trial in which half the subjects will receive OC459 and the other half placebo, before being inoculated with rhinovirus, that would normally induce a worsening of asthma symptoms i.e. an exacerbation.

Detailed description:
Asthma is the most common chronic respiratory disease, and in many countries prevalence is rising. The major morbidity, mortality and health care costs related to asthma are a result of periods of acutely increased symptomatology called 'exacerbations'. Most exacerbations are caused by rhinovirus, the virus associated with the common cold. There are few treatments to prevent and treat exacerbations, and despite these \>50% of adult asthmatics reported having an exacerbation in the last year. There is therefore a major unmet need.

Experimentally inoculating patients with asthma with rhinovirus, a methodology that has been safely used for \>15 years, induces an infection and worsening symptoms in \~85%. This model offers the possibility to investigate treatment effects on asthma exacerbations with a small number of subjects, minimising the numbers exposed to a novel drug with limited safety data. In contrast, trials of therapies powered to evaluate an effect on naturally occurring exacerbations require several hundred subjects, a long study period to capture enough events, and are significantly more expensive to carry out.

Using this model the investigators have shown that several inflammatory molecules, including prostaglandin D2 (PGD2), are significantly increased during rhinovirus-induced asthma exacerbations, with the levels of PGD2 strongly correlating with the severity of the symptoms. Moreover other studies have shown that when PGD2 binds the CRTH2 receptor, it stimulates the release of a number of inflammatory molecules also associated with asthma exacerbations. Blocking the CRTH2 receptor therefore appears an extremely promising target with potential to limit the virus-induced inflammation underpinning many asthma exacerbations.

Arms: arm 1: OC459 50mg once daily for 5 weeks arm 2: Placebo tablet once daily for 5 weeks
Interventions: intervention 1: OC459 intervention 2: Placebo intervention 3: Rhinovirus
Intervention descriptions: intervention 1: None intervention 2: None intervention 3: Inoculation with rhinovirus serotype 16

Protocol documents:
Prot_000.pdf: 
  
 
Effect of the CRTH2 Antagonist OC459 on the Response to Rhinovirus Challenge in Asthma 
 
Clinical Protocol v6.0 July 2017 
Page 1 of 38 
 
 
 
 
 
Effect of the CRTH2 antagonist OC459 on the 
Response to Rhinovirus Challenge in Asthma 
 
 
 
 
 
 
Clinical Trial Protocol 
Version 6.0, 4th July 2017 
 
 
 
 
 
Main Sponsor:  
Imperial College London 
Funder:  
Medial Research Council 
Study Coordination Centre:  Airways Disease Infection Section, 
 
National Heart & Lung Institute (NHLI), 
 
St Mary’s Campus, Imperial College London 
 
 
REC reference:  
15/LO/1666 
EudraCT reference:  
2015-002555-10 
 
 
Protocol authorised by: 
 
Name & Role 
Date 
Signature 
Prof Sebastian Johnston 
4th July 2017 
 
 


  
 
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Study Management Group 
 
Chief Investigator:  
Professor Sebastian Johnston 
 
Co-investigators:  
Dr Hugo Farne 
 
Dr David Jackson 
 
Dr Ross Walton 
 
Statistician:  
Nicholas Johnson and Professor Toby Prevost, Imperial 
Clinical Trials Unit statistics team 
 
Trial Management:  
This will be the responsibility of the Chief Investigator and Co-
Investigators. 
 
 
Study Coordination Centre 
For general queries, supply of trial documentation, and collection of data, please contact: 
 
Study Coordinator:  Dr Hugo Farne 
Address:  
Airways Disease Infection Section, National Heart & Lung Institute, 
 
Wright Fleming Building, 
 
St Mary’s Campus, Imperial College, 
 
Norfolk Place, W2 1PG, London 
Tel:  
020 7594 3751  
E-mail:  h.farne@imperial.ac.uk 
Fax:   
020 7262 8913 
 
 
 
Clinical Queries 
Clinical queries should be directed to Dr Hugo Farne, who will direct the query to the appropriate 
person. 
 
 
Sponsor 
Imperial College London is the main research Sponsor for this study. For further information 
regarding the sponsorship conditions, please contact the Clinical Trials Manager at: 
 
 
 
Joint Research Compliance Office 
5L10C, 5th Floor Lab Block 
Charing Cross Hospital 
Fulham Palace Road 
London, W6 8RF 
Tel: 0203 311 0204 
Fax: 0203 311 0203 
 
 
Funder 
This study is funded by a Medical Research Council (MRC) Biomedical Catalyst: Developmental 
Pathway Funding Scheme (DPFS) grant. Atopix Therapeutics Ltd are providing the Investigational 
Medicinal Product (IMP), OC459, and placebo free of charge in sufficient quantities for the 
purpose of the trial. 
 


  
 
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This protocol describes the Effect of the CRTH2 Antagonist OC459 on the Response to Rhinovirus Challenge in Asthma 
(ORCA) study and provides information about procedures for entering participants. The protocol should not be used as a 
guide for the treatment of other participants; every care was taken in its drafting, but corrections or amendments may be 
necessary. These will be circulated to investigators in the study, but centres entering participants for the first time are 
advised to contact the trials centre to confirm they have the most recent version.  
Problems relating to this trial should be referred, in the first instance, to the study coordination centre.  
This trial will adhere to the principles outlined in the Medicines for Human Use (Clinical Trials) Regulations 2004 (SI 
2004/1031), amended regulations (SI 2006/1928) and the International Conference on Harmonisation Good Clinical 
Practice (ICH GCP) guidelines. It will be conducted in compliance with the protocol, the Data Protection Act and other 
regulatory requirements as appropriate.  
 


  
 
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Table of Contents                                                                      Page No 
1 
INTRODUCTION 
10 
1.1 
BACKGROUND 
10 
1.2 
RATIONALE FOR CURRENT STUDY 
11 
2 
STUDY OBJECTIVES 
12 
3 
STUDY DESIGN 
13 
3.1 
STUDY OUTCOME MEASURES 
13 
4 
PARTICIPANT ENTRY 
14 
4.1 
PRE-RANDOMISATION EVALUATIONS 
14 
4.2 
INCLUSION CRITERIA 
14 
4.3 
EXCLUSION CRITERIA 
14 
4.4 
WITHDRAWAL CRITERIA 
15 
5 
RANDOMISATION AND ENROLMENT PROCEDURE 
16 
5.1 
RANDOMISATION PRACTICALITIES 
16 
5.2 
UNBLINDING 
16 
6 
TREATMENTS 
17 
6.1 
TREATMENT ARMS 
17 
7 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP) 
18 
7.1 
IMP DESCRIPTION 
18 
7.2 
STORAGE OF IMP 
18 
7.3 
DISPENSING AND ACCOUNTABILITY 
18 
7.4 
COMPLIANCE WITH TRIAL TREATMENT 
18 
7.5 
POST-TRIAL TREATMENT 
18 
8 
PHARMACOVIGILANCE 
19 
8.1 
DEFINITIONS 
19 
8.2 
CAUSALITY 
19 
8.3 
REPORTING PROCEDURES 
20 
9 
ASSESSMENT AND FOLLOW-UP 
22 
9.1 
SUMMARY OF STUDY ASSESSMENTS AND PROCEDURES 
22 
9.2 
SKIN PRICK TESTING 
22 
9.3 
BLOOD (INCLUDING VIRAL SEROLOGY) 
23 
9.4 
CLINICAL SYMPTOM SCORES (INCLUDING ACQ AND AQLQ) 
23 
9.5 
SPIROMETRY 
24 
9.6 
HISTAMINE CHALLENGE (PC20) 
25 
9.7 
EXHALED NITRIC OXIDE (FENO) 
25 
9.8 
ELECTROCARDIOGRAM (ECG) 
25 


  
 
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9.9 
URINE PREGNANCY TEST 
25 
9.10 CHEST RADIOGRAPH 
25 
9.11 NASOSORPTION 
26 
9.12 NASAL LAVAGE 
26 
9.13 NASAL SCRAPE 
26 
9.14 BRONCHOSCOPY 
26 
9.14.1 BRONCHOSCORPTION USING SYNTHETIC ABSORPTIVE MATRIX (SAM) 
27 
9.14.2 BRONCHOALVEOLAR LAVAGE (BAL) 
27 
9.14.3 BRONCHIAL BRUSHINGS 
ERROR! BOOKMARK NOT DEFINED. 
9.14.4 BRONCHIAL BIOPSY 
28 
9.15 SPUTUM INDUCTION 
28 
9.16 VIRUS INOCULATION 
29 
9.17 LOSS TO FOLLOW-UP 
29 
9.18 TRIAL CLOSURE 
29 
10 
STATISTICS AND DATA ANALYSIS 
30 
10.1 SAMPLE SIZE CALCULATION 
30 
10.2 DATA ANALYSIS 
30 
11 
QUALITY CONTROL AND QUALITY ASSURANCE PROCEDURES 
31 
11.1 RISK ASSESSMENT AND MONITORING 
31 
11.2 SERIOUS BREACHES 
31 
11.3 REPORTING 
31 
12 
REGULATORY ISSUES 
32 
12.1 CTA 
32 
12.2 ETHICS APPROVAL 
32 
12.3 CONSENT 
32 
12.4 CONFIDENTIALITY 
32 
12.5 INDEMNITY 
32 
12.6 SPONSOR 
32 
12.7 FUNDING 
32 
12.8 EXPENSES AND BENEFITS 
32 
12.9 AUDITS AND INSPECTIONS 
32 
13 
TRIAL MANAGEMENT 
33 
14 
PUBLICATION POLICY 
34 
15 
REFERENCES 
35 
APPENDICES  
37 
APPENDIX A. PHARMACY CONTACT DETAILS 
37 
APPENDIX B. PARTICIPANT DAILY DIARY CARD RECORD 
37 


  
 
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APPENDIX C. ASTHMA CONTROL QUESTIONNAIRE (ACQ) 
38 
 


  
 
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GLOSSARY OF ABBREVIATIONS 
ACQ 
Asthma Control Questionnaire 
AE 
Adverse Event 
API 
Active Pharmaceutical Ingredient 
AQLQ 
Asthma Quality of Life Questionnaire 
AR 
Adverse Reaction 
ATS 
American Thoracic Society 
BAL 
Bronchoalveolar Lavage 
BEC 
Bronchial Epithelial Cell 
BTS 
British Thoracic Society 
CRF 
Case Report Form 
CRP 
C-Reactive Protein 
CRTH2 
Chemoattractant Receptor-Homologous molecule expressed on Th2 cells 
DMP 
Data Management Plan 
ECG 
Electrocardiogram 
EDTA 
EthyleneDiamine Tetraacetic Acid 
ELF 
Epithelial Lining Fluid 
ELISA 
Enzyme-Linked ImmunoSorbent Assay 
FACS 
Fluorescence-Activated Cell Sorting 
FeNO 
Fraction of exhaled Nitric Oxide (NO) 
FEV1 
Forced Expiratory Volume in One Second 
FVC 
Forced Vital Capacity 
GCP 
Good Clinical Practice 
ICS 
Inhaled Corticosteroid 
IgE 
Immunoglobulin E 
IL 
Interleukin 
IMP 
Investigational Medicinal Product 
IUD 
Intrauterine device 
IUS 
Intrauterine system 
JRCO 
Joint Research Compliance Office 
LABA 
Long Acting Beta Agonist 
LRT 
Lower Respiratory Tract 
MDI 
Metered Dose Inhaler 
MHRA 
Medicines and Healthcare Products Regulatory Agency 
mRNA 
Messenger Ribonucleic Acid (RNA) 
MSD 
Meso Scale Discovery [a multiplex immunoassay] 
PC20  
Provocation concentration [of histamine] producing a 20% fall in FEV1 
PEF 
Peak Expiratory Flow 
PGD2 
Prostanglandin D2 
PHA 
Phytoheamagglutinin 
PPB 
Parts Per Billion 
qPCR 
Quantitative Polymerase Chain Reaction (PCR) 
REC 
Research Ethics Committee 
RV 
Rhinovirus 
RV-16 
Rhinovirus serotype 16 
SABA 
Short Acting Beta Agonist 
SAE 
Serious Adverse Event 
SAM 
Synthetic Absorptive Matrix 
SAP 
Statistical Analysis Plan 
SAR 
Serious Adverse Reaction 


  
 
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SOB 
Shortness Of Breath 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TCID50 
Tissue culture infective dose = the amount of pathogenic agent to produce a 
pathological change in 50% of cell cultures inoculated 
TLC 
Total Lung Volume 
 
KEYWORDS 
Rhinovirus, Asthma, Exacerbation, PGD2, CRTH2 
 
 
 
 


  
 
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STUDY SUMMARY 
 
TITLE Effect of OC459 on the Response to Rhinovirus Challenge in Asthma. 
DESIGN Randomised, double-blind, placebo-controlled, parallel group study. 
AIMS To evaluate if the oral CRTH2 antagonist OC459 is effective in 
preventing and/or treating rhinovirus-induced asthma exacerbations. 
OUTCOME MEASURES Primary outcome: difference in symptom scores after rhinovirus 
infection. 
Secondary outcomes: measures of health status, lung function, airway 
hyper-responsiveness, exhaled nitric oxide (FeNO) levels, virus load, and 
other laboratory parameters (e.g. differential cell counts, levels of 
cytokine/biomarker proteins and mRNA in respiratory samples). 
POPULATION 44 adults with asthma (of which 30 will be evaluable). 
ELIGIBILITY Adult subjects with moderate persistent asthma who meet the 
inclusion/exclusion criteria described. 
TREATMENT OC459 or placebo, for a total of 5 weeks per subject. 
DURATION 27 months 
 
 
FIGURE: STUDY OVERVIEW 
 
 
Week 1
2
3
4
5
6
7
8
9
Screening (2 visits)
Randomisation
Placebo
Rhinovirus inoculation
Stop trial medications
Final visit (convalescence)
OC459
Run-in period
(3 weeks)
Sampling/assessment
(2 weeks)
 


  
 
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1 
INTRODUCTION  
 
1.1 
BACKGROUND 
 
Importance of asthma and asthma exacerbations 
 
Asthma is a long-term condition affecting the airways in the lungs, causing symptoms in patients 
such as breathlessness, wheeze, tightness in the chest and cough. Asthma is the most prevalent 
chronic respiratory disease, affecting 5-10% of adults with a further 30% of school age children 
reporting wheeze [1]. Although some children will outgrow their asthma, future prevalence is 
predicted to rise: the WHO estimate 300 million people worldwide are affected by asthma and that 
this is likely to increase to 400 million cases over the next 15-20 years [2]. 
 
The major morbidity, mortality and health care costs related to asthma are a result of periods of 
acutely increased symptomatology, called ‘exacerbations’ [3]. Acute exacerbations of asthma are 
an enormous problem to adults and children: ~5-10% of asthmatics have been hospitalised with 
an exacerbation and 60% have had an exacerbation in the previous year [4]. Asthma mortality in 
the UK is ~2,000 per annum – extrapolated to Europe, this means approximately ~25,000 people 
die unnecessarily of asthma each year. The financial cost of exacerbations is estimated to be 
~50% of total expenditure on asthma care [5]. 
 
Lack of effective treatments for asthma exacerbations 
 
Currently available therapies, predominantly inhaled corticosteroids and bronchodilators, prevent 
only ~40% of asthma exacerbations [6]. These figures are in the context of clinical trials – in real 
life asthma exacerbations are even less likely to be prevented: >50% of European asthma patients 
reported having an exacerbation in the last year with >1/3 children and >1/4 adults requiring 
urgent medical care visits as a result [7]. The vast majority of asthma exacerbations therefore 
continue to occur despite use of the best available current therapies. 
 
When exacerbations do occur, treatment options are limited and have developed little in recent 
years. Treatment involves increasing doses of inhaled bronchodilators and systemic/oral steroids 
– more of the same drugs that failed to prevent the exacerbations occurring. Furthermore these 
treatments do not address the causes or mechanisms driving the exacerbations. Current 
preventive and therapeutic strategies are thus of limited efficacy and development of new 
approaches is urgently needed. 
 
Causes and mechanisms of asthma exacerbations 
 
The causes of asthma exacerbations are complex and likely to involve one or more of a number of 
factors including poor control of underlying disease, allergen exposure and viral infection. 
Respiratory viral infections appear the dominant individual precipitant as they are detected in the 
great majority of asthma exacerbations in both children (80-85%) and adults (60-80%) [8]. The 
most commonly detected viruses are human rhinoviruses (~60-80%), and it has been shown that 
experimentally infected asthmatics with rhinovirus (RV) precipitates an acute exacerbation [9]. 
 
The use of experimental RV infection has enabled the study of the mechanisms of asthma 
exacerbations. These have shown that RV infection leads to increased levels of inflammatory 
mediators in asthmatic subjects relative to healthy controls (specifically ‘type 2 cytokines’ e.g. IL-4, 
IL-5 and IL-13, as well as IL-33 and Prostaglandin D2), and that these increases correlate with the 
severity of respiratory symptoms [10]. New treatments have subsequently been developed to 
targeting those mediators, e.g. an antibody against IL-5 [11]. 


  
 
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Studies in vitro have found that the binding of Prostaglandin D2 (PGD2) to the CRTH2 receptor on 
inflammatory cells (e.g. Th2 lymphocytes, type 2 innate lymphoid cells) stimulates them to produce 
the type 2 cytokines implicated in asthma exacerbations [12, 13]. Thus treatment with a CRTH2 
antagonist represents an attractive approach to prevent RV-mediated exacerbations in asthmatic 
patients. 
 
1.2 
RATIONALE FOR CURRENT STUDY 
 
Hypothesis: This trial is designed to investigate the hypothesis that CRTH2 antagonism can 
prevent or attenuate the deterioration in symptoms in patients with asthma (i.e. asthma 
exacerbations) following rhinovirus infection. 
 
Rationale for using the RV-induced asthma exacerbation model 
 
The RV-induced asthma exacerbation model offers the ability to investigate treatment effects on 
exacerbations with a small number of volunteers, minimising the numbers exposed to a novel 
agent with limited safety data. In contrast, trials of therapies powered to evaluate an effect on 
naturally occurring exacerbations require several hundred volunteers, a long study period to 
capture enough events and are significantly more expensive to carry out. 
 
The RV-induced exacerbation model has been used extensively around the world over >15 years, 
including four studies by Professor Johnston’s group [9, 10, 14, 15], and is well-established with 
an excellent safety profile (reviewed in [16]); across the studies at Imperial there have been no 
adverse events, and no need for hospitalisation or systemic corticosteroids during the infection. 
Moreover, the RV-induced exacerbation model uniquely provides the opportunity to select a well-
defined sample population, use a uniform viral challenge, and sample repeatedly as the 
exacerbation evolves, allowing accurate dissection of the timings of responses and their 
relationship to viral load. 
 
Rationale for using the CRTH2 antagonist OC459 
 
As of August 2013, at least 12 CRTH2 antagonists were in active clinical development [17]. Of 
these, OC459 (Atopix Therapeutics Ltd) had been trialled for the longest duration in a phase 2 
study (17 weeks). In total, 637 patients across five published studies have been exposed to 
OC459 [18-22], including 482 patients with asthma, with an excellent safety record. OC459 has 
therefore been chosen for further study from the available CRTH2 antagonists due to its safety 
profile; there is no reason to think that is more or less effective than any other CRTH2 antagonist. 
 
In addition, the largest study of OC459 to date suggested that it may be effective in reducing 
exacerbations: a trend towards reduction in asthma exacerbations was noted, with a 3.8% 
incidence in the pooled dose groups compared to 7.7% on placebo (p=0.107) [22]. A dose of 25 
mg once daily was as effective as the other doses (100mg twice daily and 200mg once daily) in 
this study. 
 


  
 
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2 
STUDY OBJECTIVES 
 
Primary objective: 
 
To assess the impact of OC459 on Total Lower Respiratory Tract Symptoms Score after 
rhinovirus infection (sum of daily scores day 0 to day 14)  
 
Secondary objectives:  
 
To assess the impact of OC459 on other clinical and laboratory parameters during 
rhinovirus infection: 
o Health status (assessed by Asthma Control Questionnaire (ACQ) scores) 
o Lung function (assessed by spirometry) 
o Airway inflammation (assessed by exhaled nitric oxide (FeNO) levels) 
o Airway hyperresponsiveness (assessed as the provocation concentration of 
histamine producing a 20% fall in FEV1, or PC20) 
o Viral load (measured by quantitative Polymerase Chain Reaction (qPCR) on nasal 
lavage samples) 
 
To determine the mechanism of action of OC459, by assessing the impact of OC459 on 
the following during rhinovirus infection: 
o Eosinophil recruitment (measured by cell counts in sputum and bronchoalveolar 
lavage (BAL); eosinophil products, e.g. Eosinophil Cationic Protein (ECP), in BAL) 
o Recruitment of other immune cells (e.g. ILC2, Th2) implicated in the prostaglandin 
D2  / type 2 inflammatory pathway to the airways and/or airway mucosa (measured 
by flow cytometry to sort and count cells in BAL, serum and nasal scrapes; and 
immunohistochemistry staining of bronchial biopsies) 
o Release of inflammatory mediators implicated in the prostaglandin D2  / type 2 
inflammatory pathway (e.g. the type 2 cytokines IL-4, IL-5, IL-13; measured by 
multiplex immunoassay on nasosorption, nasal lavage, bronchosorption and BAL, 
and mRNA analysis of cells in BAL and biopsies) 
 
To assess whether OC459 is able to restore deficient innate antiviral responses 
o Production of cytokines (e.g. interferon (IFN) α and λ) and expression of cytokine 
mRNA in cells from samples (blood, BAL) and infected with rhinovirus ex vivo 
 
To assess the safety of OC459 
o Adverse event reporting, physical examinations, vital signs, and clinical laboratory 
parameters 
 
 
 


  
 
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3 
STUDY DESIGN 
 
Type of study: This is a phase 2a, randomised, double-blind, placebo-controlled, parallel group 
study. 
 
Study duration: The trial will take place over a 33 month period. On an individual basis, the trial 
will last for 9 weeks from randomisation. The trial will be considered complete when the last 
patient recruited completes the final visit. 
 
Population size: 44 asthmatic subjects. 
 
3.1 
STUDY OUTCOME MEASURES 
 
Primary outcome: 
 
Difference in Total Lower Respiratory Tract Symptoms Score after RV infection (sum of 
daily scores day 0 to day 14) between OC459 and placebo groups. 
 
Secondary outcomes:  
 
Difference between OC459 and placebo groups in the following (all measured as peak and 
change from baseline): 
o Increase in ACQ score (health status) 
o Reduction in Forced Expiratory Volume in 1 second (FEV1) or Peak Expiratory Flow 
(PEF) on spirometry 
o Increase in FeNO 
o Decrease in PC20 
o Virus load in nasal lavage – total (area under the curve) for days 2-5+7+10, as well 
as peak and difference from baseline 
o Increase in sputum eosinophilia 
o Leukocyte numbers (e.g. ILC2, Th2) in FACS-sorted BAL, serum and nasal 
scrapes, and in immunohistochemistry-stained biopsies 
o Cytokine levels (e.g. IL-4, IL-5, IL-13) in nasosorption, nasal lavage, 
bronchosorption, BAL 
o mRNA expression of same cytokines in cells in BAL and bronchial biopsies 
o In ex vivo studies, protein and mRNA levels of antiviral cytokines (e.g. IFN-α, IFN-λ) 
produced by cells from blood, BAL, and bronchial brushings in response to 
rhinovirus infection 
 
Difference in number of adverse events in OC459 and placebo groups 
 
 


  
 
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4 
PARTICIPANT ENTRY 
 
4.1 
PRE-RANDOMISATION EVALUATIONS  
 
Potential subjects for the study will be recruited from hospital outpatient clinics, GP surgeries, 
spirometry sessions/clinics, other research projects, and advertisements. They will attend the 
respiratory clinic at St Mary’s Hospital, where the outline of the study will be explained to them and 
a Participant Information Sheet (PIS) provided. They will be given the opportunity to ask questions 
and be informed that their participation is voluntary. Subjects may decide they wish to have more 
time to consider taking part in the study, in which case a follow up appointment will be made, or 
they may continue with the screening process. When a subject has had enough time to consider 
their participation in this study, and only when they have agreed to take part, they will be asked to 
read, sign and date a consent form in the presence of the investigator who will also sign the 
consent form. A copy will be kept in the research file, a copy given to the patient and a copy put 
into their medical notes. 
 
After signing a consent form, eligible subjects will undergo screening. Screening involves taking a 
medical history and examination, lung function testing, a histamine challenge, FeNO levels, skin 
prick testing, blood tests, a urine pregnancy test, and a chest radiograph to assess suitability 
against the defined inclusion and exclusion criteria. Blood tests include general screening for 
underlying illness, specifically full blood count, urea and electrolytes, liver function tests, 
coagulation, and C-reactive protein (CRP), as well as serology for rhinovirus 16 (RV-16). They will 
be matched to the inclusion/exclusion criteria, which have been maintained from previous studies 
using the rhinovirus-induced asthma exacerbation model. 
 
4.2 
INCLUSION CRITERIA 
 
 
Age 18-55 years 
 
Male or female 
 
Clinical diagnosis of asthma for at least 6 months prior to screening  
 
An Asthma Control Questionnaire (ACQ) Score >0.75  
 
Positive histamine challenge test (PC20 <8 µg/ml, or <12 µg/ml and bronchodilator 
response ≥ 12%) 
 
Worsening asthma symptoms with infection since last change in asthma therapy 
 
Positive skin prick test to common aeroallergens (e.g. animal epithelia, dust mite) 
 
Treatment comprising inhaled corticosteroids (ICS) or combination inhaler (Long-Acting 
Beta Agonist with ICS), with a daily ICS dose of at least 100mcg fluticasone or equivalent. 
 
Participant is willing for their GP to be informed of their participation. 
 
English speaker 
 
4.3 
EXCLUSION CRITERIA 
 
 
Presence of clinically significant diseases other than asthma (cardiovascular, renal, 
hepatic, gastrointestinal, haematological, pulmonary, neurological, genitourinary, 
autoimmune, endocrine, metabolic, neoplasia etc.), which, in the opinion of the 
investigator, may either put the patient at risk because of participation in the trial, or 
diseases which may influence the results of the study or the patient’s ability to take part in it 
 
Smoking history over past 12 months. 
 
Seasonal allergic rhinitis symptoms at screening or during the 3 week run-in (prior to 
rhinovirus inoculation). 


  
 
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 
Asthma exacerbation or viral illness within the previous 6 weeks or during the 3 week run-
in (prior to rhinovirus inoculation). 
 
Current or concomitant use of oral steroids, anti-leukotrienes or monoclonal antibodies. 
 
Pregnant or breast-feeding women. Patients should not be enrolled if they plan to become 
pregnant during the time of study participation (see note regarding contraception below). 
 
Contact with infants <6 months or immunocompromised persons, elderly and infirm at 
home or at work. 
 
Subjects who have known evidence of lack of adherence to medications and/or ability to 
follow physician’s recommendations. 
 
Contraception: To prevent pregnancy, male subjects with female partners of child bearing potential 
and female subjects must use at least two forms of adequate contraception for the entire duration 
of study participation. Adequate contraceptive precautions include: 
1. Established use of oral, injected or implanted hormonal methods of contraception. 
2. Placement of an intrauterine device (IUD) or intrauterine system (IUS). 
3. Barrier methods of contraception: condom or occlusive cap (diaphragm or cervical/vault caps) 
with spermicidal foam/gel/film/cream/suppository. 
4. Male sterilisation (with the appropriate post-vasectomy documentation of the absence of sperm 
in the ejaculate). 
Men whose partners are already pregnant must continue to use contraception i.e. a condom 
during the trial and for three months thereafter. The chosen contraception method(s) must be 
followed from the first Investigational Medicinal Product (IMP) administration until at least 3 
months after receiving the last dose of the IMP. 
 
Sperm donation: Male subjects must not donate sperm throughout the study and for at least 3 
months after the final dose of IMP. 
 
4.4 
WITHDRAWAL CRITERIA 
 
Participants will be free to discontinue trial medications at any point during the study without giving 
reason. Where possible, follow up will continue. If a participant chooses to withdraw consent, data 
up to the point of self-withdrawal will be included unless the participant chooses otherwise. 
 
Investigators will withdraw any subject if they are unable to tolerate the study medications, or if the 
a research investigator feels this is necessary or appropriate. In all cases, the reason for 
withdrawal must be recorded on the case report form and in the subject's medical records. 
Subjects will be closely monitored by the Clinical Research Fellow. This is defined further in the 
section on ‘Serious Adverse Events’. 
 
Subjects who are considered evaluable (i.e. confirmed rhinovirus infection) with sufficient data to 
assess the primary endpoint (i.e. complete diary cards of symptoms up to day 14 post-rhinovirus 
infection) will be included in the analysis. Subjects who are either withdrawn from the trial or who 
chose to discontinue participation and do not meet these criteria will be replaced until a sufficient 
number of evaluable subjects are reached (n=20). 
 


  
 
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5 
RANDOMISATION AND ENROLMENT PROCEDURE 
 
5.1 
RANDOMISATION PRACTICALITIES 
 
Subjects who meet the relevant criteria and provide informed consent will be randomised to either 
OC459 or placebo in a 1:1 ratio. Randomisation will occur at a baseline visit after the screening 
visit(s), and will be in blocks of four in order to balance the number of patients allocated to each 
treatment group. An unblinded randomisation list (taking into account the blocking requirements) 
will be generated by Sealed Envelope Ltd and entered into the study database. This file will be 
password protected, with unblinding instructions provided by the database development team to 
the investigators. 
 
At randomisation, the database will be interrogated and each new subject assigned the next 
sequential randomisation item on the list. The unblinded randomisation list will also be provided to 
the manufacturer, Atopix Ltd, in order to label IMP/placebo appropriately prior to dispensing to 
pharmacy. Thus the investigators, pharmacy and subjects will all be blinded. 
 
5.2 
UNBLINDING 
 
Ceasing treatment rather than unblinding will be encouraged as far as possible. However, where 
knowledge of the treatment will assist in management of acute illness / emergency treatment, 
unblinding will be supported. The decision to unblind will be made by the Chief Investigator, with 
the Trial Steering Committee convened ad hoc if necessary to support the decision. Study 
participants will be provided with a Patient Alert Card indicating the study number and a 24 hour 
emergency contact number. 
 
Unblinding will take place in line with the standard operating procedure used with the database. 
Specifically, once the decision has been made to unblind a subject, one of the study team will 
access the database and enter the subject details and study drug code. As soon as this is entered 
into the system, an email will be sent to the Trial e-mail address, the trial statistician, the Chief 
Investigator and the database team warning of a potential unblinding. The essential subject 
information will be checked against the data in the database and queries raised if they do not 
agree. The unblinding will not be actioned if there is a discrepancy between the data stored and 
the data entered. Once this information is accurately completed, a second form must be filled in to 
confirm the unblinding. The unblinded information will then be displayed, and a second email sent 
to the Trial e-mail address, the trial statistician, the Chief Investigator and the database team 
confirming the subject has been unblinded. This email will not continue details of the unblinded 
treatment. If the site unblinding account has been used, the database support team will set a new, 
pre-expired password and send new unblinding instructions to the study team. 
 
The Chief Investigator will review the cause and results of the unblinding process with the study 
team, and possibly the Trial statistician, and document/implement any follow-up actions. 
 
 


  
 
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6 
TREATMENTS 
 
6.1 
TREATMENT ARMS 
 
The Investigational Medicinal Product (IMP) used in the study will be OC459 50mg or matched 
placebo film-coated tablets. The dose of OC459/placebo will be 50mg (one tablet) to be taken 
orally, once daily, for 5 weeks in total. Both OC459 and placebo will be manufactured and 
delivered by Atopix Therapeutics Ltd at no cost to the study. These will be supplied to the 
pharmacy at St Mary’s Hospital to be stored securely. 
 
The identity of the study medications will be blinded and packaged according to the randomisation 
schedule. 
 
 


  
 
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7 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP) 
 
7.1 
IMP DESCRIPTION 
 
OC459 is a once-daily oral, highly potent and selective CRTH2 antagonist which is active on both 
the recombinant and native human receptor.  
 
The integrated safety database for OC459 includes data for a total of 754 subjects who received 
OC459 for up to 3 months with more than 500 subjects receiving doses ≥100mg daily. The overall 
incidence of adverse events was similar for those treated with OC459 (28.2%) compared to 
placebo (26.7%) and few subjects in either group had serious adverse events (0.1% and 0.5% for 
OC459 and placebo respectively). The adverse event profile after OC459 administration was 
generally comparable to that after placebo; the most common adverse event reported was 
headache (6.6% vs 5.1% for OC459 and placebo, respectively). 
  
OC459 repeat doses of up to 400mg have been studied in patients and no clear dose ordered 
effects on safety have been observed. Furthermore, no end organ toxicology or significant 
changes in clinical chemistry (including liver enzymes) have been observed in non-clinical 
toxicology studies including long term studies conducted at very high doses in the mouse (3 month 
dosing), rat (six months dosing) and the dog (12 month dosing). The known metabolites of OC459 
have been detected in at least one species including the acylglucuronide of OC459 which is the 
major metabolite in humans. 
 
7.2 
STORAGE OF IMP 
 
Clinical trial supplies will be stored below 30ºC, protected from moisture and light, in the pharmacy 
or other secure area within the investigational site. They must not be frozen or refrigerated. They 
will be stored in such a way that they cannot be mixed up or confused with other medications, be 
they clinical trial supplies or medicines for routine clinical use. In addition, returned study 
medication will not be stored with unused study medication. 
 
7.3 
DISPENSING AND ACCOUNTABILITY 
 
A record of study drug movements will be kept to maintain accountability. Only authorised 
personnel (as designated by the study delegation log) will be allowed to distribute the drugs. Any 
queries regarding supply should be directed to the study coordinator or the lead trial pharmacist 
(contact details in the Appendices). 
 
7.4 
COMPLIANCE WITH TRIAL TREATMENT 
 
Compliance will be self-reported by participants in their diary cards. In addition, we will collect and 
count returned tablets at the end of the study. Non-compliance will be defined as >20% missed 
doses as per diary cards and/or tablet count at the end of the study. Participants who are non-
compliant will be excluded from a per protocol analysis. 
 
7.5 
POST-TRIAL TREATMENT 
 
The IMP is unlicensed and therefore will not be made available to participants beyond the end of 
the study.  
 
 
 


  
 
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8 
PHARMACOVIGILANCE 
 
8.1 
DEFINITIONS   
 
Adverse Event (AE): any untoward medical occurrence in a patient or clinical trial subject 
administered a medicinal product and which does not necessarily have a causal relationship with 
this treatment. An AE can therefore be any unfavourable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease temporally associated with the use of an 
investigational medicinal product (IMP), whether or not considered related to the IMP. 
 
Adverse Reaction (AR): all untoward and unintended responses to an IMP related to any dose 
administered. All AEs judged by either the reporting investigator or the sponsor as having 
reasonable causal relationship to a medicinal product qualify as adverse reactions. The 
expression reasonable causal relationship means to convey in general that there is evidence or 
argument to suggest a causal relationship. 
 
Unexpected Adverse Reaction: an AR, the nature or severity of which is not consistent with the 
applicable product information (eg investigator’s brochure for an unapproved investigational 
product or summary of product characteristics (SmPC) for an authorised product). When the 
outcome of the adverse reaction is not consistent with the applicable product information this 
adverse reaction should be considered as unexpected. Side effects documented in the SmPC 
which occur in a more severe form than anticipated are also considered to be unexpected. 
 
Serious Adverse Event (SAE) or Serious Adverse Reaction: any untoward medical occurrence 
or effect that at any dose: 
 
Results in death. 
 
Is life-threatening – 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. 
 
Requires hospitalisation, or prolongation of existing inpatients’ hospitalisation. 
 
Results in persistent or significant disability or incapacity. 
 
Is a congenital anomaly or birth defect. 
 
Medical judgement should be exercised in deciding whether an AE/AR is serious in other 
situations. Important AE/ARs that are not immediately life-threatening or do not result in death or 
hospitalisation but may jeopardise the subject or may require intervention to prevent one of the 
other outcomes listed in the definition above, should also be considered serious. 
 
Suspected Unexpected Serious Adverse Reaction (SUSAR): any suspected adverse reaction 
related to an IMP that is both unexpected and serious.  
 
8.2 
CAUSALITY 
 
Most adverse events and adverse drug reactions that occur in this study, whether they are serious 
or not, will be expected treatment-related toxicities due to the drugs used in this study. The 
assignment of the causality should be made by the investigator responsible for the care of the 
participant using the definitions in the table below. 
 
If any doubt about the causality exists the local investigator should inform the study coordination 
centre who will notify the Chief Investigators. The pharmaceutical companies and/or other 
clinicians may be asked to advise in some cases. 
 


  
 
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In the case of discrepant views on causality between the investigator and others, all parties will 
discuss the case. In the event that no agreement is made, the MHRA will be informed of both 
points of view.  
 
Relationship 
Description 
Unrelated 
There is no evidence of any causal relationship 
Unlikely 
There is little evidence to suggest there is a causal relationship (e.g. the event 
did not occur within a reasonable time after administration of the trial 
medication). There is another reasonable explanation for the event (e.g. the 
participant’s clinical condition, other concomitant treatment). 
Possible 
There is some evidence to suggest a causal relationship (e.g. because the 
event occurs within a reasonable time after administration of the trial 
medication). However, the influence of other factors may have contributed to 
the event (e.g. the participant’s clinical condition, other concomitant 
treatments). 
Probable 
There is evidence to suggest a causal relationship and the influence of other 
factors is unlikely. 
Definitely 
There is clear evidence to suggest a causal relationship and other possible 
contributing factors can be ruled out. 
Not assessable 
There is insufficient or incomplete evidence to make a clinical judgement of the 
causal relationship. 
 
8.3 
REPORTING PROCEDURES 
 
All adverse events should be reported. Depending on the nature of the event the reporting 
procedures below should be followed. Any questions concerning adverse event reporting should 
be directed to the study coordination centre in the first instance. A flowchart is given below to aid 
in the reporting procedures. 
 
8.3.1 Non serious AR/AEs 
All such toxicities, whether expected or not, should be recorded in the toxicity section of the 
relevant case report form and sent to the study coordination centre within one month of the form 
being due.  
 
8.3.2 Serious AR/AEs 
Fatal or life threatening SAEs and SUSARs should be reported on the day that the local site is 
aware of the event. The SAE form asks for nature of event, date of onset, severity, corrective 
therapies given, outcome and causality (i.e. unrelated, unlikely, possible, probably, definitely). The 
responsible investigator should sign the causality of the event. Additional information should be 
sent within 5 days if the reaction has not resolved at the time of reporting.  
 
SAEs 
An SAE form should be completed and faxed to the study coordination centre for all SAEs within 
24 hours. 
 
SUSARs  
In the case of suspected unexpected serious adverse reactions, the staff at the site should:  
 
 
Complete the SAE case report form & send it immediately (within 24 hours, 
preferably by fax), signed and dated to the study coordination centre together with 
relevant treatment forms and anonymised copies of all relevant investigations. 


  
 
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Or 
 
Contact the study coordination centre by phone and then send the completed SAE 
form to the study coordination centre within the following 24 hours as above. 
 
The study coordination centre will notify the MHRA, REC and the Sponsor of all SUSARs 
occurring during the study according to the following timelines; fatal and life-threatening within 7 
days of notification and non-life threatening within 15 days. All investigators will be informed of all 
SUSARs occurring throughout the study. 
 
Local investigators should report any SUSARs and /or SAEs as required by their Local Research 
Ethics Committee and/or Research & Development Office. 
 
 
 
Contact details for reporting SAEs and SUSARs 
Fax: 020 7262 8913, attention Dr Hugo Farne 
Email: asthma.trial@imperial.ac.uk 
Tel: 020 7594 3764 (Mon to Fri 09.00 – 17.00) 


  
 
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9 
ASSESSMENT AND FOLLOW-UP  
 
9.1 
SUMMARY OF STUDY ASSESSMENTS AND PROCEDURES 
 
After screening and providing informed consent, eligible participants will undergo baseline 
investigations and randomisation, followed by a 21 day run-in period on placebo or OC459. 
Towards the end of this period they will have a baseline bronchoscopy and other investigations. At 
the end of the run-in period, subjects will have a further set of investigations prior to rhinovirus 
inoculation the same day. Regular sampling and assessments will take place as indicated whilst 
treatment administration continues uninterrupted for a further 14 days, for a total of 5 consecutive 
treatment weeks. Trial medications are then stopped (day 14). Subjects will attend for a final 
follow-up visit 4 weeks later with convalescence investigations (day 42). Table 1 summarises 
subject visits and procedures during the study. Details of the individual study procedures are listed 
below. 
 
Study participants will be seen frequently during the study period, daily if they wish. They have 
details to contact the Clinical Research Fellow and Clinical Research Nurse, and will be offered 
daily telephone contact. In this way participants will be assessed regularly by the investigating 
team and any adverse events detected rapidly. When the study is completed, subjects will return 
to the care of their GP (they will not be routinely followed-up by the study group). 
 
 
Screening 
visit 1 
Screening 
visit 2 
Visit 1 
(baseline) 
Visit 2 
Visit 3 
Visit 4 
Visit 5 
Visit 6 
Visit 7 
Visit 8 
Visit 9 
Visit 10 
Visit 11 
Study day 
 
 
-21 
-9 
-8 
0 
2 
3 
4 
5 
7 
10 
42 
Ideal weekday 
 
 
F 
W 
T 
F 
Su 
M 
T 
W 
F 
M 
F 
Visit length 
30 
min 
60 
min 
60 
min 
45 
min 
3  
hr 
60 
min 
45 
min 
45 
min 
45 
min 
3 
hr 
60 
min 
45 
min 
30 
min 
Skin prick test 
X 
 
 
 
 
 
 
 
 
 
 
 
 
Viral serology 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
Asthma Control Questionnaire (ACQ) 
X 
 
X 
X 
 
X 
 
 
 
 
 
X 
X 
Spirometry (in clinic) 
X 
 
X 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
Histamine challenge (PC20) 
 
X 
X 
X 
 
 
 
 
 
 
X 
 
 
Exhaled nitric oxide (FeNO) 
 
X 
X 
X 
 
X 
 
X 
 
X 
X 
X 
X 
Blood tests 
 
X 
X 
 
X 
 
 
X 
 
X 
 
X 
X 
ECG 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Urine pregnancy test 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Chest radiograph 
 
X 
 
 
 
 
 
 
 
 
 
 
 
IMP or placebo administration 
 
 
Daily from day -21                                                  day 14 (then stop) 
 
Nasosorption 
 
 
X 
 
 
X 
X 
X 
X 
X 
X 
X 
 
Nasal lavage 
 
 
X 
 
 
X 
X 
X 
X 
X 
X 
X 
 
Nasal scrape 
 
 
X 
 
X 
 
 
X 
 
X 
 
 
 
Bronchoscopy (bronchosorption, BAL, 
brushings, biopsies) 
 
 
 
 
X 
 
 
 
 
X 
 
 
 
Sputum induction 
 
 
X 
X 
 
 
 
 
X 
 
X 
 
 
Virus inoculation 
 
 
 
 
 
X 
 
 
 
 
 
 
 
Symptom diaries including spirometry 
and medication 
 
 
Daily at home during study period 
Spirometry (portable, at home) 
 
 
Daily at home during study period 
Table 1: Summary of study visits and key procedures 
 
9.2 
SKIN PRICK TESTING 
 
Atopic status will be determined by skin prick testing to a range of 10 common aeroallergens. 
Positive histamine / negative diluent controls will be included. 
- Mixed grass pollen  
 
- House dust mite (Der P)  


  
 
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- Silver birch  
 
 
- Mugwort 
- 3 trees 
 
 
 
- Aspergillus fumigatus 
- Cat dander  
 
 
- Cladasporium herbarum 
- Dog hair  
 
 
 
- Alternaria alternata  
 
 
 
 
 
≥ 1 positive reaction (wheal 3 mm greater than the negative control) will be considered diagnostic 
of atopy. 
 
9.3 
BLOOD (INCLUDING VIRAL SEROLOGY) 
 
Screening visit blood (in tubes with ethylenediamine tetraacetic acid (EDTA), citrate, or heparin) 
will be taken for full blood count, renal function, liver function tests, glucose, clotting, CRP, vitamin 
D and total IgE. These will be processed in the Haematology and Chemical Pathology 
Laboratories of St Mary’s Hospital.  
 
Blood will also be taken on study visits as per Table 1; the specific samples taken on each visit are 
outlined in Table 2. Serology will be performed at screening and day 42 for detection of antibodies 
to rhinovirus-16 (RV-16). Serum will also be used to count cell types in the circulating blood using 
flow cytometry, and to assess the immune response to rhinovirus ex vivo. 
 
Study day 
Full blood count 
(EDTA 4.0mL) 
Clotting 
(Citrate 2.7mL) 
Biochem + IgE 
(Heparin 3.5mL) 
Serum 
Screening  
x1 
x1 
x2 
6mL 
Day -21 
 
 
 
80mL 
Day -7 
x1 
x1 
x2 
80mL 
Day 3 
x1 
x1 
x2 
10mL 
Day 5 
x1 
x1 
x2 
80mL 
Day 10 
x1 
x1 
x2 
10mL 
Day 42 
x1 
x1 
x2 
6mL 
Total (mL) 
24.0mL 
16.2mL 
42.0mL 
292mL 
  Table 2: Overview of blood, plasma, and serum requirements (mL) 
 
9.4 
CLINICAL SYMPTOM SCORES (INCLUDING ACQ)  
 
Patients will be required to keep a diary card of both upper and lower respiratory tract symptoms 
during the study period (see Appendices). The diary cards will be commenced 21 days prior to 
inoculation (on day -21) to derive a baseline symptom score. Symptom scores will be kept daily for 
6 weeks post-inoculation until the convalescent visit. Spirometry measurements and drug 
compliance will be recorded on the same diary cards. 
 
Upper Respiratory Tract Symptoms 
The diary card of upper respiratory tract symptoms will be started 21 days prior to inoculation. A 
total upper respiratory clinical symptom score will be derived using a scale of 0 to 3 (absent, mild, 
moderate and severe) for each of the following eight respiratory symptoms: sneezing, headache, 
malaise, fever/chills, nasal discharge, nasal obstruction, sore throat and cough, according to 
previously established criteria [23]. Symptoms will be recorded at the same time of day and before 
any procedures such as bronchoscopy are performed. An example is shown below: 
 
Symptom 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 6 
Day 7 
Sneezing 
 
 
 
 
 
 
 
Headache 
 
 
 
 
 
 
 
Malaise 
 
 
 
 
 
 
 


  
 
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Fever/chills 
 
 
 
 
 
 
 
Nasal discharge 
 
 
 
 
 
 
 
Nasal obstruction 
 
 
 
 
 
 
 
Sore throat 
 
 
 
 
 
 
 
Cough 
 
 
 
 
 
 
 
Total score 
 
 
 
 
 
 
 
0 = absent, 1 = mild, 2 = moderate, 3 = severe 
 
The maximum clinical severity score on any one day would be 24. Individual symptom scores will 
be accumulated over the six-day period from the onset of the cold and the recording on the day 
prior to inoculation subtracted. Thus, for a patient who has a score of zero on day -1 prior to 
inoculation, the maximum cumulative score for the six days is 144. 
 
Lower Respiratory Tract Symptoms 
A diary card of lower respiratory tract symptoms will be completed with a scoring system as 
outlined below: 
 
Symptom 
Day 1 
Day 2 
Day 3 
Day 4 
Day 5 
Day 6 
Day 7 
Cough on waking 
 
 
 
 
 
 
 
Wheeze on waking 
 
 
 
 
 
 
 
Daytime cough 
 
 
 
 
 
 
 
Daytime wheeze 
 
 
 
 
 
 
 
Daytime breathlessness 
 
 
 
 
 
 
 
Nocturnal cough, 
wheeze, breathlessness 
 
 
 
 
 
 
 
Total score 
 
 
 
 
 
 
 
0 = absent, 1 = mild, 2 = moderate, 3 = severe 
 
An exacerbation of asthma is defined as an increase in the total respiratory score of at least 2 
points over baseline on 2 consecutive days. 
 
Asthma Control Questionnaire (ACQ) 
The ACQ is a standard measure of disease severity and combines symptom scores with objective 
parameters such as FEV1 predicted and medication use. The ACQ should be interviewer 
administered. Further details are in the appendices. ACQ scoring will be assessed by study team 
on study visits as per Table 1. 
 
9.5 
SPIROMETRY 
 
Spirometry will be performed using a MicroLab spirometer (CareFusion, Kent, UK) on study visits 
as per Table 1. At the screening visit measurements of Forced Expiratory Volume in 1 second 
(FEV1) and Forced Vital Capacity (FVC) will be made: 
 
Posture must be consistent during a study, either standing or sitting, with no breathing 
limitation. 
 
The subject should breathe in to total lung capacity. A good tight seal by the lips round the 
mouthpiece is essential. The subject should then exhale forcibly into the spirometer, 
blowing as hard as possible and continue to residual volume. 
 
The best value of 3 attempts will be recorded. 
 
The predicted or reference values for lung function measurements are those recommended by the 
Report Working Party for the European Community for Coal and Steel. Also incorporated are the 
recommendations of the British Thoracic Society (BTS) and the Association of Respiratory 
Technicians and Physiologists. 


  
 
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Reversibility: At screening, spirometry will be repeated 15 minutes after the administration of 
200μg salbutamol via metered dose inhaler (MDI) and volumatic spacer to assess reversibility. A 
cut-off point for significant reversibility of 12% of FEV1 predicted and 200ml will be taken as 
supportive of a diagnosis of asthma. Clinic spirometry will be carried out as part of the protocol for 
induced sputum. 
 
Home spirometry: The subjects will be provided with a portable PiKo-1 spirometer (nSpire Health, 
Hertford, UK) to carry out daily measurements of FEV1, FVC and PEF at home. At the baseline 
visit the subjects will be instructed in the use of the spirometer and will commence measurements 
to gain familiarity with its use. Measurements will be continued daily for the 6 weeks after 
inoculation. 
 
9.6 
HISTAMINE CHALLENGE (PC20) 
 
Histamine challenge (maximum concentration 16 mg/mL) will be performed according to American 
Thoracic Society (ATS) guidelines by using the 2 minute tidal breathing method [24]. The 
provocative concentration of histamine causing a 20% reduction in FEV1 (PC20) is then calculated. 
 
9.7 
EXHALED NITRIC OXIDE (FENO) 
 
Measurement of FeNO levels will be performed using a NIOX VERO device (Aerocrine, Solna, 
Sweden) with single use patient filters on study visits as per Table 1, and prior to all other 
assessments. The patient should not eat or drink or participate in any strenuous physical activity 
for at least one hour prior to testing. The measurement of FeNO will be performed according to 
American Thoracic Society (ATS) guidelines [25]: 
 
Patients should be seated comfortably  
 
Explain test procedure to patient, emphasisng that it is a relaxed  manoeuvre 
 
Instruct patient to  
1. Empty their lungs by breathing out thoroughly 
2. Close their lips around the mouthpiece on the patient filter so that no air leakage 
occurs 
3. Inhale deeply through the patient filter to Total Lung Capacity (TLC) 
4. Exhale slowly through the filter using the display as a guide to the required 
exhalation pressure and duration 
The instrument will analyse the sample and generate a result in parts per billion (PPB) in 
approximately one minute. 
 
9.8 
ELECTROCARDIOGRAM (ECG) 
 
All patients eligible to participate in the study will have a single electrocardiogram (ECG), as part 
of routine assessment for any medical abnormalities. 
 
9.9 
URINE PREGNANCY TEST 
 
All female patients eligible to participate in the study will have a single urine pregnancy test. 
 
9.10 
CHEST RADIOGRAPH 
 
All patients eligible to participate in the study will have a single chest radiograph performed prior to 
their bronchoscopy, as per our local research bronchoscopy guidelines. The radiation dose 
associated with a single chest radiograph is minimal (0.02mSv, equivalent to 3 days natural 


  
 
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background radiation) and the risk of adverse effects is negligible (risk of inducing a cancer 
~1:1,000,000). 
 
9.11 
NASOSORPTION 
 
This minimally invasive technique samples nasal (i.e. upper airway) mucosal lining fluid and offers 
potential benefits over nasal lavage including improved sensitivity through avoidance of significant 
analyte dilution (inherent with lavage). Nasosorption will be performed on study visits as per Table 
1, prior to bronchoscopy and nasal lavage to avoid contamination by the local anaesthetic 
(bronchoscopy) or saline (nasal lavage) introduced into the nose.  
 
Two strips of Synthetic Absorptive Matrix (SAM) (Leukosorb, Pall Life Sciences, Hampshire, UK) 
measuring 7mm x 35mm will be placed inside the participant’s nostrils for 2 minutes to obtain 
samples of nasal lining fluid. This is a painless, minimally invasive procedure that will not require 
any local anaesthetic. 
 
Each piece of SAM will be placed in a single labelled Spin-X Centrifuge Tube with Filter (Sigma-
Aldrich Cat no: CLS8161) containing an assay buffer. This will be transported to the lab on ice for 
initial processing and transferred to -80°C pending analysis for proteins and cytokines by multiplex 
immunoassay (e.g. using the Meso Scale Device, MSD). 
 
9.12 
NASAL LAVAGE 
 
Nasal lavage is performed on study visits as per Table 1 using the following technique: 
 
With subjects head extended and soft palate closed, up to 10mL of sterile normal saline is 
instilled into each nostril using a Pasteur pipette. 
 
Subjects then gently blow their nose into a sterile universal container or sputum pot. 
 
Lavage fluid is stored on ice, vortexed and aliquoted for storage at -80°C. 
 
Nasal lavage fluid will be analysed to quantify the degree of viral shedding and determine viral 
load. Nasal lavage samples pre- and post-rhinovirus inoculation will also be tested for all common 
respiratory viruses by a PCR panel to exclude coincidental infections. Where possible, nasal 
lavage fluid will also be analysed for proteins and cytokines by multiplex immunoassay. 
 
9.13 
NASAL SCRAPE 
 
Up to four nasal scrapes will be performed on study visits as per Table 1 to obtain nasal epithelial 
cells. All nasal scrapes will be performed after the collection of nasosorption and nasal lavage 
samples. They will be collected using the following technique: 
 
A careful examination of the nose is made to identify the nasal mucosa on the inferior 
turbinate. A plastic nasal curette (Rhinoprobe, Arlington Scientific) is advanced into the 
nostril until the tip is placed on the surface of the inferior turbinate. Using a gentle scraping 
motion a tissue sample is collected. This method of sampling is almost painless, and has 
been extensively employed in adults and infants. 
 
Nasal scrapes will be placed in labelled sterile scrape tubes with culture media or flow 
buffer,  for culture and flow cytometry respectively. 
 
9.14 
BRONCHOSCOPY 
 
Bronchoscopies will be performed on study visits as per Table 1 in the endoscopy suite at St 
Mary’s Hospital, in accordance with British Thoracic Society (BTS) guidelines [26]: 


  
 
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 
Subjects will sign a consent form on entering the study and will sign a separate consent 
form for each bronchoscopy. The procedure will be explained during the assessment stage 
and subjects will be given written information specifically regarding bronchoscopies in 
addition to the Participant Information Sheet. 
 
Subjects will fast for six hours prior to the procedure. 
 
Resuscitation equipment (for intubation, ECG monitoring and defibrillation) and necessary 
drugs (salbutamol, theophylline, adrenaline, hydrocortisone) will be available in the 
bronchoscopy room. 
 
Premedication will be given including: 
- Nebulised salbutamol 2.5mg prior to bronchoscopy 
- Sedation with midazolam (2-10mg) and/or fentanyl (25-200mcg) as necessary 
- Lignocaine spray (10%) and solutions (1%) for topical anaesthesia. The total dose will 
not exceed 7mg/kg or 400mg in total. 
 
Supplemental oxygen at a rate of 2Lmin-1 is given via a nasal cannula and oxygen 
saturations and heart rate are monitored with a pulse oximeter continuously. Intravenous 
access will be mandatory in all cases. 
 
The subject will be monitored during the bronchoscopy by a separate nurse and a second 
suitably qualified physician will be present to act as an advocate. 
 
The following samples will be collected in this order: 
1. Bronchosorption using Synthetic Absorptive Matrix (SAM)  
2. Bronchial brushings  
3. Bronchoalveolar lavage (BAL)  
4. Bronchial biopsy 
At the second bronchoscopy, sampling should be performed on the contralateral side to 
the analogous procedures done at the first bronchoscopy. 
 
All adverse events (pain, bleeding, hypoxia etc) will be recorded. 
 
Subjects will be observed for 1-2 hours, nil by mouth, after the procedure. 
 
Transportation will be arranged as subjects should not drive on the day of the procedure. 
 
All subjects will have a contact telephone number on discharge. 
 
9.14.1 Bronchoscorption using Synthetic Absorptive Matrix (SAM) 
Bronchosorption (Mucosal Diagnostics, Hunt Developments Ltd, Midhurst, UK) is a 
technique to sample bronchial mucosal lining fluid. The main benefit of this technique is the 
measurement of previously undetectable mediators through avoidance of the significant 
analyte dilution associated with bronchoalveolar lavage.  
 
The bronchosorption device is passed down the operating port of the bronchoscope. 
The distal end of the inner probe incorporates a folded strip of Leukosorb (Pall Life 
Sciences, Hampshire, UK) measuring 1.8mm x 30mm which are placed on either the 
right, or left (chosen at random) lower lobe bronchial mucosa for 30 seconds.  
 
Following sampling, the bronchosorption device is withdrawn back into its sheath and 
the complete device is removed from the bronchoscope. 
 
The sampling end of the probe is cut off and treated in an identical way to the 
nasosorption strips of Leukosorb.  
Proteins and cytokines will be measured in the samples (e.g. IL-4, IL-5 and IL-13 by 
multiplex immunoassay such as MesoScale Discovery (MSD) Th1/Th2 7-plex). 
 
9.14.2 Bronchial brushings 
Up to six bronchial brushings will be taken from the lower or middle lobe sub-segmental 
bronchi with a standard disposable cytology brush at each bronchoscopy: 
 
Deploy each brush at segmental or sub-segmental carina. Advance 1-2cm but keep the 
brush in sight. Gently brush the side-wall avoiding any bleeding. If a brush has blood on 


  
 
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Clinical Protocol v6.0 July 2017 
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it, dispose and flush the bronchoscopic channel with a small volume (2-5ml of sterile 
saline before proceeding with additional brushes. 
 
Place each brush in a 2ml screw cap tube containing 1mL RNAse-free PBS on ice. Cut 
the wire and close the cap over the brush. 
 
Store brushing tubes on ice until further processing. 
Cells are removed from the brushes and split into samples to be either cultured for ex vivo 
studies, or used for RNA extraction and analysis. 
 
9.14.3 Bronchoalveolar lavage (BAL) 
 
The bronchoscope is gently inserted into one segmental bronchus of either the right or 
left middle lobes.  
 
BAL is performed by instillation of sterile physiological (0.9%) saline at room 
temperature in 30ml aliquots to a total of 180-240ml, aiming for a return of ~100ml. 
 
The BAL fluid is collected into a plastic chamber and transferred to polypropylene 
tubes for transport on ice to the laboratory. 
Samples will be centrifuged to separate the cells and supernatant. The cells will be subject 
to mRNA analysis and flow cytometry, to identify and separate the different inflammatory 
cell phenotypes. The supernatants will be analysed for proteins and cytokines (where they 
are present in sufficient concentrations). 
 
9.14.4 Bronchial biopsy 
Six bronchial biopsies will be taken from the segmental and sub-segmental bronchi of the 
right lower and middle lobe at each bronchoscopy, using Keymed 2mm biopsy channel 
cupped and fenestrated biopsy forceps [FB-19C-1 (1111065)]. Four biopsies will be placed 
in 4% paraformaldehyde for preparation of paraffin blocks. Immunohistochemistry staining 
will be used to detect different phenotypes of inflammatory cells. The remaining two 
biopsies will be used for mRNA analysis. 
 
9.15 
SPUTUM INDUCTION 
 
Sputum induction will be performed on study visits as per Table 1, in accordance with ERS 
guidelines[27].  
 
Subjects will be pre-medicated with 200μg salbutamol via metered dose inhaler and large 
volume spacer and baseline FEV1 measured after 10 minutes. 
 
If the subject’s FEV1 is ≥60% of predicted, the following procedure should be used:  
o Administer 3-4.5% hypertonic saline via nebulizer.  
o Perform 2-5 minutes of inhalation then stop inhalation and check FEV1 reading. 
o Repeat this process 3 times (i.e. a maximum of 20 minutes).  
 
If the subjects FEV1 <60% of predicted, the following should be used:  
o Use normal saline (0.9%) initially. 
o Perform 30 seconds of inhalation then stop inhalation and check FEV1 reading.  
o Repeat this process for 1 min and then 2 mins of inhalation.  
o Repeat this process with 3% saline with inhalations of 30 secs, 1 min and 2 mins.  
o Repeat this process with 3-4.5% hypertonic saline for 30 secs, 1, 2, 4, and 8 mins.  
 
In either case if the subject’s FEV1 drops by 20% at any time, or if the subject experiences 
excessive coughing, bronchoconstriction, hyperventilation, dyspnoea, dizziness, nausea 
and vomiting, or an increased wheeze, the induction should be stopped immediately and 
rescue therapy given. 
 
Sputum plugs are selected from saliva by macroscopic inspection of the sample. An aliquot is 
analysed, unprocessed, for virus load by quantitative PCR. The remaining sample is processed 


  
 
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Clinical Protocol v6.0 July 2017 
Page 29 of 38 
and centrifuged to separate the cells, which are then stained and counted for differential cell 
counts. 
 
9.16 
VIRUS INOCULATION 
 
Details of the production and testing of the rhinovirus serotype 16 (RV-16) inoculum have been 
previously published [28]. This inoculum was prepared under the supervision of the Director of the 
Medical Research Council Common Cold Unit and was fully safety tested for adventitious agents 
according to published safety guidelines. It has been stored in individual unopened aliquots in the 
applicants -80°C freezer since production. It has been used in previous studies at high dose [9, 
28-30] and low dose [31] over a time span >15 years. There have been no serious adverse events 
in any of these studies. 
 
Subjects will be inoculated on a single occasion with stored inoculum, described above, divided 
equally between the two nostrils. A total dose of 100 TCID50 (tissue culture infective dose; the 
amount of a pathogenic agent that will produce pathological change in 50% of cell cultures 
inoculated) is used. This is done slowly with sufficient interval between each spray to ensure 
maximum contact time between the spray and the nasal and pharyngeal mucosa. Subjects will be 
asked not to swallow during the procedure to ensure maximal pharyngeal contact. Subjects will 
also be asked to sniff at each actuation to encourage delivery of particles of inoculum to the lower 
airway. Identical methods have been used in our previous studies in COPD and asthmatic 
subjects. 
 
9.17 
LOSS TO FOLLOW-UP 
 
Participants who are lost to follow up will not be replaced if they were evaluable (i.e. confirmed 
rhinovirus infection) and there is sufficient data to assess the primary outcome (i.e. complete diary 
card of symptoms to day 14 post-rhinovirus infection). Subjects who are lost to follow up and do 
not meet these criteria will be replaced until a sufficient number of evaluable subjects are reached 
(n=20). 
 
9.18 
TRIAL CLOSURE 
 
The trial will be considered complete when the last patient recruited completes the final visit. 
 
 


  
 
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Clinical Protocol v6.0 July 2017 
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10 
STATISTICS AND DATA ANALYSIS 
 
10.1 
SAMPLE SIZE CALCULATION 
 
The sample size determination is based on the following assumptions: 
 
Type I error probability α = 5% 
 
Effect size = (µ1-µ0)/σ, where  
µi = mean PEP in group i=(i=1: OC459, i=0: placebo), 
σ = standard deviation of PEP, and  
PEP is primary end-point (=total daily lower respiratory symptom scores over d0 to d14). 
 
Based on a previously completed trial with similar design conducted at the same study site [10], σ 
is estimated to be 21.15 and the effect size equal to 22.21, yielding n=15 evaluable subjects per 
treatment group at 80% power. This is grossed-up for 80% rhinovirus inoculation success and 
adjusted for expected drop-outs to yield 22 enrolled patients per treatment group. 
 
10.2 
DATA ANALYSIS 
 
Subjects completing the study protocol will be defined as infected or un-infected (neutralising 
antibody to RV-16 at 6 weeks post inoculation at a titre <1:4 and no detection of RV-16 by qPCR 
in nasal lavage, sputum or BAL at any point after inoculation). Only subjects with confirmed 
rhinovirus infection will be included in statistical analysis. 
 
Quantitative assessments of symptom scores, lung function, exhaled nitric oxide (FeNO), airway 
hyperresponsiveness (histamine challenge, PC20), virus load, inflammatory cell numbers and 
inflammatory markers will be compared within subjects to determine differences between baseline 
and during infection. Within-subject differences will be analyzed using ANOVA and 2-tailed paired 
Student’s t-tests or Wilcoxon signed rank test as appropriate. 
 
Differences between the patients taking OC459 and placebo group will be analyzed using 
unpaired parametric or non-parametric analysis as appropriate at each phase of the study. 
Correlations between (but not limited to) the following will be examined using Spearman’s rank 
correlations to investigate possible causal relationships: 
 
illness severity (symptoms, lung function, airway hyperresponsiveness) 
 
virus load 
 
inflammatory cell counts 
 
inflammatory markers  
 
gene expression 
 
Data and all appropriate documentation will be stored for a minimum of 10 years after the 
completion of the study, including the follow-up period. 


  
 
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11 
QUALITY CONTROL AND QUALITY ASSURANCE PROCEDURES 
 
11.1 
RISK ASSESSMENT AND MONITORING 
 
Regular monitoring will be performed by the Joint Research Compliance Office (JRCO) Clinical 
Trial Monitor at Imperial College, who will monitor the study regularly based on a completed risk 
assessment by the sponsor. The study will be monitored according to Good Clinical Practice 
(GCP). Data will be evaluated for compliance with the protocol and accuracy in relation to source 
documents. Following written standard operating procedures, the monitors will verify that the 
clinical trial is conducted and data are generated, documented and reported in compliance with the 
protocol, GCP and the applicable regulatory requirements. 
 
11.2 
SERIOUS BREACHES 
 
The Medicines for Human Use (Clinical Trials) Regulations contain a requirement for the 
notification of "serious breaches" to the MHRA within 7 days of the Sponsor becoming aware of 
the breach. 
 
A serious breach is defined as “A breach of GCP or the trial protocol which is likely to affect to a 
significant degree –  
 
(a) the safety or physical or mental integrity of the subjects of the trial; or 
(b) the scientific value of the trial”. 
 
In the event that a serious breach is suspected the Sponsor must be contacted within 1 working 
day. In collaboration with the C.I., the serious breach will be reviewed by the Sponsor and, if 
appropriate, the Sponsor will report it to the REC committee, Regulatory authority and the NHS 
host organisation within seven calendar days. 
 
11.3 
REPORTING 
 
The Chief Investigator shall submit once a year throughout the clinical trial, or on request, an 
Annual Progress Report to the Research Ethics Committee, host organisation, and Sponsor.  In 
addition, an End of Trial notification and final report will be submitted to the MHRA, the REC, host 
organisation and Sponsor. 
 
 


  
 
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12 
REGULATORY ISSUES 
 
12.1 
CTA 
Clinical Trials Authorisation will be obtained from the UK Competent Authority, the Medicines & 
Healthcare products Regulatory Agency (MHRA), prior to starting recruitment. 
 
 
12.2 
ETHICS APPROVAL 
The Chief Investigator will obtain approval from a Research Ethics Committee. The study must be 
submitted for Site Specific Assessment (SSA) at each participating NHS Trust. The Study 
Coordination Centre will require a copy of the Trust R&D approval letter before accepting 
participants into the study. The study will be conducted in accordance with the recommendations 
for physicians involved in research on human subjects adopted by the 18th World Medical 
Assembly, Helsinki 1964 and later revisions. 
 
12.3 
CONSENT  
Consent to enter the study must be sought from each participant only after a full explanation has 
been given, an information leaflet offered and time allowed for consideration. Signed participant 
consent should be obtained. The right of the participant to refuse to participate without giving 
reasons must be respected. After the participant has entered the trial the clinician remains free to 
give alternative treatment to that specified in the protocol at any stage if he/she feels it is in the 
participant’s best interest, but the reasons for doing so should be recorded. In these cases the 
participants remain within the study for the purposes of follow-up and data analysis. All 
participants are free to withdraw at any time from the protocol treatment without giving reasons 
and without prejudicing further treatment. 
 
12.4 
CONFIDENTIALITY 
Participants’ identification data will be required for the registration process. The Study 
Coordination Centre will preserve the confidentiality of participants taking part in the study and is 
registered under the Data Protection Act. 
 
12.5 
INDEMNITY 
Imperial College London holds negligent harm and non-negligent harm insurance policies which 
apply to this study. 
 
12.6 
SPONSOR 
Imperial College London will act as the main Sponsor for this study. Delegated responsibilities will 
be assigned to the NHS trusts taking part in this study. 
 
12.7 
FUNDING 
The Medical Research Council (MRC) are funding this study. 
 
12.8 
EXPENSES AND BENEFITS 
Participants in the study will have their travel expenses refunded. They will also be given a 
donation of £1,500 to compensate for time and inconvenience of taking part in the study. If 
subjects drop out, they will receive a pro rata amount: £300 if after the first bronchoscopy; £500 if 
after rhinovirus inoculation; £1,000 if after the second bronchoscopy; and £1,500 on completion. 
 
12.9 
AUDITS AND INSPECTIONS  
The study may be subject to inspection and audit by Imperial College London under their remit as 
Sponsor, the Study Coordination Centre and other regulatory bodies to ensure adherence to GCP.  
 


  
 
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Clinical Protocol v6.0 July 2017 
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13 
TRIAL MANAGEMENT 
 
The day-to-day management of the trial will be co-ordinated by Dr Hugo Farne (Clinical Research 
Fellow), with close support from Dr David Jackson, Dr Ross Walton, and Professor Sebastian 
Johnston.  
 
A Trial Steering Committee will be appointed and will be responsible for overseeing the progress 
of the trial. This will incorporate the activities of a Data Monitoring Committee. The Trial Steering 
Committee will include members of the research team, a patient representative, and a consultant 
in Respiratory medicine independent of the study group. An independent statistician will be invited 
as required. The Trial Steering Committee will meet every six months (and additionally on an ad 
hoc basis as required) to review the data, oversee the safety of subjects in the trial, and ensure 
that no events have occurred that would slow the progression of the trial. The minutes from these 
meetings will be stored in the Trial Management Folder. 


  
 
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14 
PUBLICATION POLICY 
 
It is expected that after analysis the data from this study will be widely distributed in the medical 
and scientific community. Facilitated with presentations at local, national and international 
meetings, we hope to publish widely in the medical literature. In addition, we have an excellent 
media department at Imperial College and publicise research that has public interest when it is 
published. 
 


  
 
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15 
REFERENCES 
 
1. 
Asher, M.I., et al., Worldwide time trends in the prevalence of symptoms of asthma, allergic 
rhinoconjunctivitis, and eczema in childhood: ISAAC Phases One and Three repeat 
multicountry cross-sectional surveys. Lancet, 2006. 368(9537): p. 733-43. 
2. 
Organization., W.H., Global surveillance, prevention and control of chronic respiratory 
diseases: a comprehensive approach. 2007, Geneva. 
3. 
Murray, C.J. and A.D. Lopez, Alternative projections of mortality and disability by cause 
1990-2020: Global Burden of Disease Study. Lancet, 1997. 349(9064): p. 1498-504. 
4. 
Rabe, K.F., et al., Clinical management of asthma in 1999: the Asthma Insights and Reality 
in Europe (AIRE) study. Eur Respir J, 2000. 16(5): p. 802-7. 
5. 
Weiss, K.B. and S.D. Sullivan, The health economics of asthma and rhinitis. I. Assessing 
the economic impact. J Allergy Clin Immunol, 2001. 107(1): p. 3-8. 
6. 
Pauwels, R.A., et al., Effect of inhaled formoterol and budesonide on exacerbations of 
asthma. Formoterol and Corticosteroids Establishing Therapy (FACET) International Study 
Group. N Engl J Med, 1997. 337(20): p. 1405-11. 
7. 
Papadopoulos, N.G., et al., Mechanisms of virus-induced asthma exacerbations: state-of-
the-art. A GA2LEN and InterAirways document. Allergy, 2007. 62(5): p. 457-70. 
8. 
Singanayagam, A., et al., Viruses exacerbating chronic pulmonary disease: the role of 
immune modulation. BMC Med, 2012. 10: p. 27. 
9. 
Message, S.D., et al., Rhinovirus-induced lower respiratory illness is increased in asthma 
and related to virus load and Th1/2 cytokine and IL-10 production. Proc Natl Acad Sci U S 
A, 2008. 105(36): p. 13562-7. 
10. 
Jackson, D.J., et al., IL-33-dependent type 2 inflammation during rhinovirus-induced 
asthma exacerbations in vivo. Am J Respir Crit Care Med, 2014. 190(12): p. 1373-82. 
11. 
Castro, M., et al., Reslizumab for inadequately controlled asthma with elevated blood 
eosinophil counts: results from two multicentre, parallel, double-blind, randomised, 
placebo-controlled, phase 3 trials. Lancet Respir Med, 2015. 
12. 
Xue, L., A. Barrow, and R. Pettipher, Interaction between prostaglandin D and 
chemoattractant receptor-homologous molecule expressed on Th2 cells mediates cytokine 
production by Th2 lymphocytes in response to activated mast cells. Clin Exp Immunol, 
2009. 156(1): p. 126-33. 
13. 
Xue, L., et al., Prostaglandin D2 activates group 2 innate lymphoid cells through 
chemoattractant receptor-homologous molecule expressed on TH2 cells. J Allergy Clin 
Immunol, 2014. 133(4): p. 1184-94. 
14. 
Mallia, P., et al., Experimental rhinovirus infection as a human model of chronic obstructive 
pulmonary disease exacerbation. Am J Respir Crit Care Med, 2011. 183(6): p. 734-42. 
15. 
Mallia, P., et al., Rhinovirus infection induces degradation of antimicrobial peptides and 
secondary bacterial infection in chronic obstructive pulmonary disease. Am J Respir Crit 
Care Med, 2012. 186(11): p. 1117-24. 
16. 
Del Vecchio, A.M., et al., Utility of animal and in vivo experimental infection of humans with 
rhinoviruses in the development of therapeutic agents for viral exacerbations of asthma 
and chronic obstructive pulmonary disease. Pulm Pharmacol Ther, 2015. 30: p. 32-43. 
17. 
Norman, P., Update on the status of DP2 receptor antagonists; from proof of concept 
through clinical failures to promising new drugs. Expert Opin Investig Drugs, 2014. 23(1): p. 
55-66. 
18. 
Barnes, N., et al., A randomized, double-blind, placebo-controlled study of the CRTH2 
antagonist OC000459 in moderate persistent asthma. Clin Exp Allergy, 2012. 42(1): p. 38-
48. 


  
 
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19. 
Horak, F., et al., The CRTH2 antagonist OC000459 reduces nasal and ocular symptoms in 
allergic subjects exposed to grass pollen, a randomised, placebo-controlled, double-blind 
trial. Allergy, 2012. 67(12): p. 1572-9. 
20. 
Singh, D., et al., Inhibition of the asthmatic allergen challenge response by the CRTH2 
antagonist OC000459. Eur Respir J, 2013. 41(1): p. 46-52. 
21. 
Straumann, A., et al., Anti-eosinophil activity and clinical efficacy of the CRTH2 antagonist 
OC000459 in eosinophilic esophagitis. Allergy, 2013. 68(3): p. 375-85. 
22. 
Pettipher, R., et al., Heightened response of eosinophilic asthmatic patients to the CRTH2 
antagonist OC000459. Allergy, 2014. 69(9): p. 1223-32. 
23. 
Jackson, G.G., et al., Transmission of the common cold to volunteers under controlled 
conditions. I. The common cold as a clinical entity. AMA Arch Intern Med, 1958. 101(2): p. 
267-78. 
24. 
Crapo, R.O., et al., Guidelines for methacholine and exercise challenge testing-1999. This 
official statement of the American Thoracic Society was adopted by the ATS Board of 
Directors, July 1999. Am J Respir Crit Care Med, 2000. 161(1): p. 309-29. 
25. 
Society, A.T., Recommendations for standardized procedures for the on-line and off-line 
measurement of exhaled lower respiratory nitric oxide and nasal nitric oxide in adults and 
children-1999. This official statement of the American Thoracic Society was adopted by the 
ATS Board of Directors, July 1999. Am J Respir Crit Care Med, 1999. 160(6): p. 2104-17. 
26. 
Du Rand, I.A., et al., British Thoracic Society guideline for diagnostic flexible bronchoscopy 
in adults: accredited by NICE. Thorax, 2013. 68 Suppl 1: p. i1-i44. 
27. 
Paggiaro, P.L., et al., Sputum induction. Eur Respir J Suppl, 2002. 37: p. 3s-8s. 
28. 
Bardin, P.G., et al., Experimental rhinovirus infection in volunteers. Eur Respir J, 1996. 
9(11): p. 2250-5. 
29. 
Fraenkel, D.J., et al., Lower airways inflammation during rhinovirus colds in normal and in 
asthmatic subjects. Am J Respir Crit Care Med, 1995. 151(3 Pt 1): p. 879-86. 
30. 
Seymour, M.L., et al., Rhinovirus infection increases 5-lipoxygenase and cyclooxygenase-2 
in bronchial biopsy specimens from nonatopic subjects. J Infect Dis, 2002. 185(4): p. 540-
4. 
31. 
Mallia, P., et al., An experimental model of rhinovirus induced chronic obstructive 
pulmonary disease exacerbations: a pilot study. Respir Res, 2006. 7: p. 116. 
 


  
 
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APPENDICES 
 
Appendix A. Pharmacy Contact Details 
 
Severine Rey 
Lead trial pharmacy clinical trials technician 
Outpatient Pharmacy Department 
St Mary’s Hospital 
Imperial College Healthcare NHS Trust 
Praed Street 
London W2 1PG 
Telephone:  
020 3312 7643 
Fax: 
 
020 3312 6708 
 
 
Appendix B. Participant daily diary card record 
 
 
 
 


  
 
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Appendix C. Asthma Control Questionnaire (ACQ) 
 
1. On average, during the past week, how often were you woken by your asthma during the night? 
 
0  
Never 
1  
Hardly ever 
2  
A few minutes 
3  
Several times 
4  
Many times 
5  
A great many times 
6  
Unable to sleep because of asthma 
 
2. On average, during the past week, how bad were your asthma symptoms when you woke up in the morning? 
 
0  
No symptoms 
1  
Very mild symptoms 
2  
Mild symptoms 
3  
Moderate symptoms 
4  
Quite severe symptoms 
5  
Severe symptoms 
6  
Very severe symptoms 
 
3. In general, during the past week, how limited were you in your activities because of your asthma? 
 
0  
Not limited at all 
1  
Very slightly limited 
2  
Slightly limited 
3  
Moderately limited 
4  
Very limited 
5  
Extremely limited 
6  
Totally limited 
 
4. In general, during the past week, how much shortness of breath did you experience because of your asthma? 
 
0  
None 
1  
A very little 
2  
A little 
3  
A moderate amount 
4  
Quite a lot 
5  
A great deal 
6  
A very great deal 
 
5. In general, during the past week, how much of the time did you wheeze?  
 
0  
Not at all 
1  
Hardly any of the time 
2  
A little of the time 
3  
A moderate amount of the time 
4  
A lot of the time 
5  
Most of the time 
6  
All the time 
 
6. On average, during the past week, how many puffs of short-acting bronchodilator (eg. Ventolin) have you used each day? 
 
0  
None 
1  
1-2 puffs most days 
2  
3-4 puffs most days 
3  
5-8 puffs most days 
4  
9-12 puffs most days 
5  
13-16 puffs most days 
6  
More than 16 puffs most days 
 
7. FEV1 pre-bronchodilator:  
 
0  
>95% predicted 
1  
95-90% 
2  
89-80% 
3  
79-70% 
4  
69-60% 
5  
59-50% 
6  
<50% predicted 

SAP_001.pdf: 


ORCA_Asthma_SAP_v1.0 
 
page 2 of 35 
Effect of the CRTH2 antagonist OC459 on the Response to Rhinovirus Challenge in 
Asthma  
15/LO/1666 
Table of Contents 
2. 
Study Objectives / Hypotheses Testing 
3 
2.1. 
Primary Objective 
3 
2.2. 
Secondary Objectives 
3 
3. 
Study Endpoints 
4 
3.1. 
Primary Endpoint Variable 
4 
3.2. 
Secondary Endpoint Variables 
4 
4. 
Background/Introduction 
5 
4.1. 
Introduction 
5 
4.2. 
Study Design 
5 
4.3. 
Study Population 
6 
4.4. 
Sample Size 
7 
4.5. 
Treatment Groups 
7 
4.6. 
Randomisation 
7 
4.7. 
Withdrawal and Replacement 
8 
4.8. 
Unblinding 
8 
4.9. 
Schedule of Time and Events 
9 
4.10. 
Study Baseline 
10 
5. 
Analysis Sets 
10 
5.1. 
Safety analysis set 
10 
5.2. 
Full analysis set 
10 
5.3. 
Per protocol analysis set 
10 
5.4. 
Extended analysis set 
10 
6. 
Variables of Analysis 
11 
6.1. 
Baseline Demographic Variables 
11 
6.2. 
Primary Analysis Variable 
11 
6.3. 
Secondary Analysis Variables 
11 
6.3.1. 
Secondary Efficacy Variables 
11 
6.3.2. 
Secondary Mechanistic Variables 
12 
6.3.3. 
Safety Analysis Variables 
12 
6.4. 
Exploratory Analysis Variables 
12 
7. 
Statistical Methodology 
14 
7.1. 
Baseline Demographics 
14 
7.2. 
Primary Analysis 
14 
7.3. 
Secondary Analysis 
14 
7.3.1. 
Secondary Efficacy Analysis 
14 


ORCA_Asthma_SAP_v1.0 
 
page 3 of 35 
7.3.2. 
Secondary Mechanistic Analysis 
15 
7.3.3. 
Correlation testing between Secondary Outcomes 
16 
7.3.4. 
Safety Analysis 
16 
7.4. 
Exploratory Analysis 
17 
7.4.1. 
Exploratory Mechanistic Testing 
17 
7.4.2. 
Exploratory Testing For Antiviral Response 
17 
7.4.3. 
Exploratory Correlation Testing 
18 
7.4.4. 
Further Testing For Outcome Variables 
18 
7.5. 
Missing Data and Withdrawn Subjects 
20 
7.6. 
Data Handling & Transformation 
21 
7.7. 
Software of Analysis 
21 
APPENDICIES 
 
2. 
Study Objectives / Hypotheses Testing  
 
 
 
 
2.1. Primary Objective 
To assess the impact of OC459 on Total Lower Respiratory Tract Symptoms Score after rhinovirus 
infection (sum of daily scores day 0 to day 14) 
 
2.2. Secondary Objectives 
 
To assess the impact of OC459 on other clinical and laboratory parameters during 
rhinovirus infection in asthma: 
o Health status  
o Lung function  
o Airway inflammation 
o Airway hyper-responsiveness  
o Viral load 
 
To determine the mechanism of action of OC459, by assessing the impact of OC459 on 
the following during rhinovirus infection: 
o Eosinophil recruitment 
o Recruitment of other immune cells (e.g. ILC2, Th2) implicated in the 
prostaglandin D2 / type 2 inflammatory pathway to the airways and/or airway 
mucosa 
o Release of inflammatory mediators implicated in the prostaglandin D2 / type 2 
inflammatory pathway (e.g. the type 2 cytokines IL-4, IL-5, IL-13) 
 
To assess the safety of OC459 
 
Exploratory - To assess whether OC459 is able to restore deficient innate antiviral 
responses in asthma 


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3. 
Study Endpoints 
3.1. Primary Endpoint Variable 
 Total Lower Respiratory Tract Symptoms Score after rhinovirus infection (sum of daily 
scores day 0 to day 14) 
 
3.2. Secondary Endpoint Variables 
 To assess the impact of OC459 on other clinical and laboratory parameters during 
rhinovirus infection in asthma: 
o Upper Respiratory Tract Symptoms Scores from D0 to D14 
o Corrected Lower Respiratory Tract Symptoms Scores from D0 to D14 
o Asthma Control Questionnaire (ACQ-6) scores  
o Spirometry – FEV1, FVC, PEFR 
o Exhaled nitric oxide (FeNO) levels 
o Provocation concentration [of histamine] producing a 20% fall in FEV1 (PC20) 
o Viral load (measured by qPCR) 
 To determine the mechanism of action of OC459, by assessing the impact of OC459 on 
the following during rhinovirus infection in asthma: 
o cell counts in sputum and bronchoalveolar lavage (BAL)  
o immune cells (e.g. ILC2, Th2) in BAL, blood and nasal scrapes, measured by flow 
cytometry 
o immune cells in bronchial biopsies, measured by immunohistochemistry staining 
o soluble mediators in the upper and lower airways, measured by multiplex 
immunoassay on nasosorption, nasal lavage, bronchosorption and BAL samples 
 To assess the safety of OC459 
o Adverse event reporting 
o physical examinations 
o vital signs 
o clinical laboratory parameters 
 Exploratory - To assess whether OC459 is able to restore deficient innate antiviral 
responses 
o Production of antiviral interferons (IFN), e.g. β and λ by cells from blood and 
bronchial brushings following infection with respiratory viruses ex vivo 
o expression of mRNA for interferon and a panel of interferon-stimulated genes in 
cells from the same ex vivo studies 
 
 
 


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4. 
Background/Introduction  
4.1. Introduction 
Asthma is a long-term condition affecting the airways in the lungs, causing symptoms in patients 
such as breathlessness, wheeze, tightness in the chest and cough. Asthma is the most prevalent 
chronic respiratory disease, affecting 5-10% of adults with a further 30% of school age children 
reporting wheeze. Although some children will outgrow their asthma, future prevalence is 
predicted to rise: the WHO estimate 300 million people worldwide are affected by asthma and 
that this is likely to increase to 400 million cases over the next 15-20 years [2].  
The major morbidity, mortality and health care costs related to asthma are a result of periods of 
acutely increased symptomatology, called ‘exacerbations’. Acute exacerbations of asthma are an 
enormous problem to adults and children: ~5-10% of asthmatics have been hospitalised with an 
exacerbation and 60% have had an exacerbation in the previous year. Asthma mortality in the 
UK is ~1,000 per annum – extrapolated to Europe, this means approximately ~11,000 people die 
unnecessarily of asthma each year. The financial cost of exacerbations is estimated to be ~50% 
of total expenditure on asthma care. 
As of August 2013, at least 12 CRTH2 antagonists were in active clinical development. Of these, 
OC459 (Atopix Therapeutics Ltd) had been trialled for the longest duration in a phase 2 study (17 
weeks). In total, 637 patients across five published studies have been exposed to OC459, 
including 482 patients with asthma, with an excellent safety record. OC459 has therefore been 
chosen for further study from the available CRTH2 antagonists due to its safety profile; there is 
no reason to think that it is more or less effective than any other CRTH2 antagonist.  
In addition, the largest study of OC459 to date suggested that it may be effective in reducing 
exacerbations: a trend towards reduction in asthma exacerbations was noted, with a 3.8% 
incidence in the pooled dose groups compared to 7.7% on placebo (p=0.107). A dose of 25 mg 
once daily was as effective as the other doses (100mg twice daily and 200mg once daily) in this 
study. 
Based on the above; this trial is designed to investigate the hypothesis that CRTH2 antagonism 
can prevent or attenuate the deterioration in symptoms in patients with asthma (i.e. asthma 
exacerbations) following rhinovirus infection. 
 
4.2. Study Design 
This is a randomised, double-blind, placebo-controlled, parallel group phase 2a study which will 
take place over a 36 month period. On an individual basis, the trial will last for 9 weeks from 
randomisation.  
 
The trial will be considered complete when the last patient recruited completes the final visit. 
 
 
 
 


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4.3. Study Population 
English speaking, male or female patients between the age of 18-55 who have had a Clinical 
diagnosis of asthma for at least 6 months prior to screening will be deemed eligible for the trial if 
they satisfy the following additional Inclusion Criteria: 
 
An Asthma Control Questionnaire (ACQ-6) Score >0.75  
 
Positive histamine challenge test (PC20 <8 μg/ml, or <12 μg/ml and bronchodilator 
response ≥ 12%)  
 
Worsening asthma symptoms with infection since last change in asthma therapy  
 
Positive skin prick test to common aeroallergens (e.g. animal epithelia, dust mite)  
 
Treatment comprising inhaled corticosteroids (ICS) or combination inhaler (Long-Acting 
Beta Agonist with ICS), with a daily ICS dose of at least 100mcg fluticasone or equivalent.  
 Participant is willing for their GP to be informed of their participation.  
Patients must also not fall under any of the following exclusion criteria: 
 
Presence of clinically significant diseases other than asthma (cardiovascular, renal, 
hepatic, gastrointestinal, haematological, pulmonary, neurological, genitourinary, 
autoimmune, endocrine, metabolic, neoplasia etc.), which, in the opinion of the 
investigator, may either put the patient at risk because of participation in the trial, or 
diseases which may influence the results of the study or the patient’s ability to take part 
in it  
 
Smoking history over past 12 months.  
 
Seasonal allergic rhinitis symptoms at screening or during the 3 week run-in (prior to 
rhinovirus inoculation).  
 
Asthma exacerbation or viral illness within the previous 6 weeks or during the 3 week 
run-in (prior to rhinovirus inoculation).  
 
Current or concomitant use of oral steroids, anti-leukotrienes or monoclonal antibodies.  
 
Pregnant or breast-feeding women. Patients should not be enrolled if they plan to 
become pregnant during the time of study.  
 
Contact with infants <6 months or immunocompromised persons, elderly and infirm at 
home or at work.  
 
Subjects who have known evidence of lack of adherence to medications and/or ability to 
follow physician’s recommendations.  


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Whilst not listed within the trial protocol; an additional test was carried out to assess previous 
exposure to rhinovirus 16, as evidenced by the presence of serum neutralising antibodies to 
rhinovirus 16 at screening. 
 
Subjects who meet the relevant criteria and provide informed consent are deemed eligible for 
randomisation to the trial. 
 
4.4. Sample Size 
An overall population size of 44 asthmatic subjects was based on the following assumptions: 
 
 Type I error probability α = 5% 
 
 Effect size = (μ1-μ0)/σ 
o 
μi = mean PEP in group i=(i=1: OC459, i=0: placebo), 
o 
σ = standard deviation of PEP 
 
 PEP is primary end-point (=total daily lower respiratory symptom scores over D0 to D14, 
potential maximum of 315). 
 
Based on a previously completed trial with similar design conducted at the same study site [10], 
σ is estimated to be 21.15 and the effect size equal to 22.21, yielding n=15 evaluable subjects 
per treatment group at 80% power. This is grossed-up for 80% rhinovirus inoculation success and 
adjusted for expected drop-outs to yield 22 enrolled patients per treatment group. 
 
 
4.5. Treatment Groups 
 
OC459: 50mg to be taken orally as one tablet, once daily, for 5 weeks in total. 
 
Placebo:  to be taken orally as one tablet, once daily, for 5 weeks in total. 
 
4.6. Randomisation 
Subjects who meet the relevant criteria and provide informed consent will be randomised to 
either OC459 or placebo in a 1:1 ratio. Randomisation will occur at a baseline visit after the 
screening visit(s), and will be in blocks of four in order to balance the number of patients 
allocated to each treatment group. A dummy unblinded randomisation list (taking into account 
the blocking requirements) will be generated by the trial statistician and tested. Once tested, a 
statistician working independent to the trial will create the final list for entering into the study 
database. This file will be password protected, with unblinding instructions provided by the 
database development team to the investigators. 
At randomisation, the database will be interrogated and each new subject assigned the next 
sequential randomisation item on the list. The unblinded randomisation list will also be provided 
to the manufacturer, Atopix Ltd, in order to label IMP/placebo appropriately prior to dispensing 
to pharmacy. Thus the investigators, pharmacy and subjects will all be blinded. 


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4.7. Withdrawal and Replacement 
Subjects who are considered evaluable (confirmed rhinovirus infection) and have completed at 
least 14 days post inoculation with RV-16 will be included in the analysis. Subjects lost to follow 
up will not be replaced if they were evaluable with sufficient data to assess the primary 
outcome. Subjects who are either withdrawn from the trial or who chose to discontinue 
participation and do not meet these criteria will be replaced until a sufficient number of 
evaluable subjects are reached (n=30).  
 
4.8. Unblinding 
Ceasing treatment rather than unblinding will be encouraged as far as possible. However, where 
knowledge of the treatment will assist in management of acute illness / emergency treatment, 
unblinding will be supported. The decision to unblind will be made by the Chief Investigator, 
with the Trial Steering Committee convened ad hoc if necessary to support the decision. Study 
participants will be provided with a Patient Alert Card indicating the study number and a 24 hour 
emergency contact number. 
 
Unblinding will take place in line with the standard operating procedure used with the database. 
Specifically, once the decision has been made to unblind a subject, one of the study team will 
access the database and enter the subject details and study drug code. As soon as this is entered 
into the system, an email will be sent to the Trial e-mail address, the trial statistician, the Chief 
Investigator and the database team warning of a potential unblinding. The essential subject 
information will be checked against the data in the database and queries raised if they do not 
agree. The unblinding will not be actioned if there is a discrepancy between the data stored and 
the data entered. Once this information is accurately completed, a second form must be filled in 
to confirm the unblinding. The unblinded information will then be displayed, and a second email 
sent to the Trial e-mail address, the trial statistician, the Chief Investigator and the database 
team confirming the subject has been unblinded. This email will not continue details of the 
unblinded treatment. If the site unblinding account has been used, the database support team 
will set a new, pre-expired password and send new unblinding instructions to the study team. 
 
The Chief Investigator will review the cause and results of the unblinding process with the study 
team, and possibly the Trial statistician, and document/implement any follow-up actions.  
 


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4.9. Schedule of Time and Events 
Table 1: Schedule of Visits and Events 
 
Table 2: Schedule of Treatment and Events 
Day
-
-21
-9 -8
0 2 3 4 5 7 10
42
Symptom scores
Lower respiratory
Daily at home throughout study period
Upper respiratory
Daily at home throughout study period
ACQ-6
X
X
X
X
Lung function
Home PEFR
Daily at home throughout study period
Home FEV1
Daily at home throughout study period
Clinic FEV1
X
X
X
X X X X X X
X
X
Clinic FEV1:FVC ratio
X
X
X
X X X X X X
X
X
Lung inflammation
FeNO
X
X
X
X
X
X X
X
X
PC20
X
X
X
X
Blood samples
Blood eosinophils
X
X
X
X
X
X
X
Nasal samples
X
X
X X X X X X
X
Virus load
X
X
X X X X X X
X
Soluble mediators
X
X
X X X X X X
X
Bronchial samples
Soluble mediators
X
X
Flow cytometry
Blood cell counts
X
X
X
X
Nasal cell counts
X
X
X
X
Bronchial cell counts
X
X
3 weeks
2 weeks
4 weeks
OC459
Placebo
Follow up
Screening
Randomisation
Rhinovirus
inoculation
Stop trial
medications
 


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4.10. Study Baseline 
For Primary and Secondary endpoint assessments, baseline will be determined as the last 
measurement taken prior to virus inoculation i.e. Day 0 (ACQ-6, home PEFR, home FEV1, clinic FEV1, 
clinic FVC, FeNO, virus load, soluble mediators) where available. If/where this is missing, the value on 
the previous visit will be used, for example, laboratory bloods and PC20 (for laboratory bloods, the 
sample on day -8 will be taken as a baseline, for PC20, the result from day -9 will be taken). 
 
Demographic baseline variables will be taken at Screening, Day -21 and Day 0 (where available). 
 
5. 
Analysis Sets 
5.1. Safety analysis set 
The safety analysis set includes all subjects who were randomized and received at least one dose of 
study medication. 
 
5.2. Full analysis set 
The full analysis set (FAS) includes subjects who: 
 
Are randomised into the study (D-21) 
 
Have been inoculated with the rhinovirus serotype 16 (RV-16) challenge virus (D0) 
 
Have confirmed RV-16 infection, defined as either (i) positive RV-16 PCR in nasal lavage 
at any time after inoculation (D0) or (ii) seroconversion (positive antibodies to RV-16 at a 
titre of at least 1:4 at the final study visit) 
 
Have completed at least 14 days post inoculation with RV-16. 
These subjects are defined as ‘evaluable’ and form the basis of the power calculation. 
The FAS will be used to assess the primary objective and will be used to analyse all efficacy endpoints 
plus any post-infection mechanistic outcomes.  
5.3. Per protocol analysis set 
The per protocol (PP) analysis set includes subjects in the FAS without any major protocol deviations 
that could confound the interpretation of analyses conducted on the FAS. Major protocol deviations 
will be defined prior to database lock and without knowing the treatment of individual cases. Patients 
taking less than 80% of the prescribed medication will be excluded from the PP set. Patients that fail to 
provide a minimum 80% of endpoint data will also be excluded from per-protocol analysis from that 
particular endpoint. 
Any per-protocol analysis would be limited to the primary and efficacy/symptom-based secondary 
endpoints as defined in Sections 6.2 & 6.3.1. 
 
5.4. Extended analysis set 
The extended analysis set expands on the FAS by including any subject who completed the study, 
regardless of whether they had confirmed RV-16 infection. The set will be used to investigate pre-
infection mechanistic effects of OC459 and will also be used for any ex-vivo analyses.


ORCA_Asthma_SAP_v1.0 
 
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6. 
Variables of Analysis 
6.1. Baseline Demographic Variables 
 
Gender 
 
Ethnicity 
 
Age (at enrolment) 
 
Height, weight & BMI 
 
Duration of asthma 
 
History of previous oral steroids 
 
Skin prick test: (a) which allergens (b) cumulative wheal size in mm2 
 
Smoking history +/- pack years 
 
Baseline asthma medications (this includes the ICS dose) 
 
Values of the following at screening and, in supplementary tables, at randomisation (day 
-21) and at visit 4 (day 0): 
o 
ACQ-6 
o 
FEV1 + reversibility 
o 
FVC 
o 
FeNO 
o 
PC20 
o 
Blood eosinophils 
o 
Serum IgE 
o 
Serum vitamin D (screening and randomisation only) 
 
6.2. Primary Analysis Variable 
 The primary endpoint will consist of the sum of uncorrected lower respiratory symptom 
scores from day 0 to day 14. Daily scores will be out of 21 (seven symptom questions 
rated from zero to three) creating a potential maximum total score of 315. 
 
6.3. Secondary Analysis Variables 
6.3.1. 
Secondary Efficacy Variables 
 
Upper respiratory symptom score 
 
Lower respiratory symptom score 
 
ACQ-6 Score 
 
PEFR 
 
FEV1  


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 
FVC 
 
FeNO  
 
PC20  
 
Virus load (via qPCR) 
6.3.2. 
Secondary Mechanistic Variables 
1. Blood & Sputum eosinophilia  
2. Leukocyte numbers (e.g. ILC2, Th2) in BAL, blood and nasal scrapes measured by flow 
cytometry 
3. Leukocyte numbers in immunohistochemistry-stained bronchial biopsies: 
o 
CRTH2+ cells (cells/mm2 tissue) 
 
In the submucosa 
 
In the epithelium 
o 
EG2+ activated eosinophils (cells/mm2 tissue) 
 
In the submucosa 
 
In the epithelium 
4. Soluble Mediators (nasal and bronchial): 
o 
Prostaglandin D2 
o 
Th2 cytokines (3): IL-4, IL-5, IL-13 
o 
Th1 cytokines (2): IFN-γ, IL-12p70 
o 
Th17 cytokines (4): IL-17A, IL-17F, GM-CSF, IL-22 
o 
Chemokines (7): Eotaxin, Eotaxin 3, IP-10, MDC, TARC, MIP-1α, MIP-1β  
o 
Pro-inflammatory markers (4): IL-6, IL-8, IL-1β, TNF-α  
o 
Cytokines that can trigger type 2 inflammation (3): IL-25/-17E, IL-33, TSLP 
o 
Innate antiviral interferons (2): IFN-α, IL-29/IFN-λ1 
o 
Additional (6): IL-9, IL-10, IL-15, IL-18, IL-23, MIP-3α 
 
6.3.3. 
Safety Analysis Variables 
 
Adverse events (recorded in free text; not coded) 
 
Serious adverse events 
 
Concomitant medications (recorded in free text, not coded) 
 
Vital signs (temperature, pulse, systolic/diastolic blood pressure, oxygen saturation 
levels, respiratory rate) 
 
Laboratory blood tests (full blood count, renal function, liver enzymes, clotting) 
 
6.4. Exploratory Analysis Variables 
Exploratory analysis will take place across all cell counts obtained above using flow cytometry 
and bronchial biopsy from blood, nasal and bronchoscopic samples: A comprehensive list of 
which can be found below: 
 Total viable cells 
 Total CRTH2+ cells 


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page 13 of 35 
 Th2 cells (defined as CD3+CD4+CRTH2+) 
 
Of which are also GATA3+  
 Eosinophils (defined as CD66b+CD16-) 
 Neutrophils (defined as CD66b+CD16+) 
 Mast cells (defined as FcERI+Lin-CRTH2-CD117+) 
 Basophils (defined as FcERI+Lin-CRTH2+CD117-) 
 ILC2 cells (defined as Lin-CD127+FcERI-CRTH2+) 
 ILC1 cells (defined as Lin-CD127+FcERI-CRTH2-CD117-) 
 ILC3 cells (defined as Lin-CD127+FcERI-CRTH2-CD117+) 
Blood samples will be assessed at day -21, day -8, day 5, day 42; nasal samples at day -21, day -8, 
day 5 and bronchoscopic samples at day -8, day 5 (counted by flow cytometry) 
Further exploratory analysis will take place on cells obtained from bronchial biopsies from day -8 
and day 5 by immunohistochemistry: 
 CRTH2+ cells (cells/mm2 tissue) 
 
In the submucosa 
 
In the epithelium 
 EG2+ activated eosinophils (cells/mm2 tissue) 
 
In the submucosa 
 
In the epithelium 
Values for ACQ-6 (D-9, D10) will also be analysed. 
 
In addition, exploratory analysis will take place to investigate antiviral response. This will consist 
of the following variables: 
 
Protein levels of IFN-β, -λ1 and –λ2/3 in supernatants of cells taken from subjects and 
infected with virus ex vivo. 
 
mRNA levels of IFN-β, -λ1 and –λ2/3 a panel of interferon stimulated genes (Mx1, RIG-I, 
OAS, viperin, IP-10, PKC) in cell lysates from the same ex vivo experiments. 
 


ORCA_Asthma_SAP_v1.0 
 
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7. 
Statistical Methodology 
7.1. 
Baseline Demographics 
Baseline demographic variables and other relevant clinical baseline characteristics as defined in 
Section 6.1 will be summarised for each treatment group. 
Summaries of continuous variables will be presented as means and standard deviations if data is 
consistent with that from a normal population distribution, and as medians and inter-quartile 
ranges for data that is inconsistent. Categorical variables will be presented as frequencies and 
percentages. 
 
7.2. 
Primary Analysis 
Uncorrected lower respiratory symptom scores collected from D0 to D14 will be aggregated to 
create an overall total for each patient. Where patient scores are missing, data will be imputed 
based upon rules set within Section 7.5. Patients missing ≥20% of score data will be excluded 
from any additional per-protocol analyses. 
Data will be presented using summary statistics per treatment group and will include median 
and interquartile range (IQR). A supplementary plot showing individual measurements for both 
groups with a box & whisker overlay showing median and IQR will be produced. 
Primary analysis to test for difference between the mean aggregated scores of the two 
treatment groups will be carried out using Mann-Whitney U tests. Differences will be considered 
significant at P < 0.05. All P values are two-sided. 
An additional secondary analysis will take place correcting the scores for bronchoscopy-related 
symptoms. Full methodology will be clarified within Section 7.3.1 below. 
 
7.3. 
Secondary Analysis 
7.3.1. 
Secondary Efficacy Analysis 
Lower respiratory symptom scores collected from D0 to D14 will be corrected for the effects of 
bronchoscopy by subtracting the baseline symptoms value (mean average of values from D-13 to 
D-9) from the two days affected by bronchoscopy symptoms (day -8 and day -7). The adjusted 
residual symptom scores for day -8 and day -7 (which we assume to be purely bronchoscopy 
related) would then be subtracted from day 5 and day 6 to complete the correction. 
Analysis to test for difference between the mean aggregated corrected scores of the two 
treatment groups will be carried out using Mann-Whitney U tests.  
The effect of the drug on asthma exacerbations (as defined by change from day 0 to peak 
infection in symptoms, lung function, and measures of lung inflammation) will be using 
presented summary statistics per treatment group and will include median and interquartile 
range (IQR). Within-subject differences will be analysed using ANOVA and 2-tailed paired 
Student’s t-tests or Wilcoxon signed rank test in the event where data is not consistent with a 
normal population distribution Analysis to determine the difference in effect of OC459 and 
placebo will be performed using unpaired (two-sample) t-test or Mann-Whitney U tests for data 
not consistent with a normal population distribution at each phase of the study.  
Differences will be considered significant at P < 0.05 within a hierarchy of the primary outcome, 
and interpreted more cautiously for secondary efficacy outcomes, (likewise more so for 


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page 15 of 35 
exploratory outcomes). Patients missing ≥20% of endpoint data will be excluded from any 
additional per-protocol analyses. Analyses that are conducted using the Per Protocol analysis set 
that are in agreement with conclusions derived from the Full analysis set will be more strongly 
interpreted than those in disagreement . All P values are two-sided. 
 
 
7.3.2. Secondary Mechanistic Analysis 
Soluble Mediators: 
A total of 32 mediators in nasal samples (taken at day -21, 0, 2, 3, 4, 5, 7, 10) and bronchoscopic 
samples (taken at day -8, day 5) will be measured for each subject. 
Values of soluble mediators obtained from bronchial samples will be presented using summary 
statistics per treatment group and visit. An additional table will summarise per treatment group 
(for each mediator) the difference in values between mediators taken at D-8 and D5. Analysis to 
investigate any difference in effect of OC459 and placebo for each mediator will be performed 
using the difference in values via unpaired (two-sample) t-test or Mann-Whitney U tests for non-
parametric data. 
Values of soluble mediators obtained from nasal samples will be presented using summary 
statistics per treatment group and visit. An additional table will summarize per treatment group 
(for each mediator) the difference between D-21 and peak value (the highest value measured 
between D0 and D10). 
Taking the difference between D-21 and peak value, analysis to determine the difference in 
effect of OC459 and placebo for each mediator will be performed using unpaired (two-sample) t-
test or Mann-Whitney U tests for non-parametric data.  
Differences will be considered significant at P < 0.05 within a hierarchy of the primary outcome, 
and interpreted more cautiously for secondary efficacy outcomes, (likewise more so for 
exploratory outcomes). All P values are two-sided. 
 
Leukocyte numbers: 
Values of leukocyte numbers obtained from bronchial samples / flow cytometry will be 
presented using summary statistics per treatment group and visit. An additional table will 
summarise per treatment group (for each cell type) the difference in values between samples 
taken at D-8 and D5. Analysis to investigate any difference in effect of OC459 and placebo for 
each cell type will be performed using the difference in values via unpaired (two-sample) t-test 
or Mann-Whitney U tests for non-parametric data. 
 
 
Eosinophil counts: 
Values of blood and sputum eosinophilia will be presented using summary statistics per 
treatment group and visit. Analysis to investigate any difference in effect of OC459 and placebo 
for blood and sputum eosinophilia will be performed using the difference in values via unpaired 
(two-sample) t-test or Mann-Whitney U tests for non-parametric data. 
 


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page 16 of 35 
 
 
 
7.3.3. 
Correlation testing between Secondary Outcomes 
Potential causal relationships between outcome variables will be investigated using Spearman’s 
rank correlations. Tests will be carried out between (but not limited to) the following: 
 
Baseline values of: 
 
ACQ-6 (day 0) 
 
FeNO (day 0) 
 
Blood eosinophils (day -8) 
 
Serum IgE (day -8) 
 
PC20 (day -9) 
 
Skin prick test (from screening) 
o # of allergens 
o Sum of wheal diameters (mm) 
 
Soluble mediators 
 
vs 
Peak and AUC values of: 
 
Lower respiratory symptom score 
 
Upper respiratory symptom score 
 
Virus load 
 
Morning PEFR (peak decline) 
 
Morning FEV1 (peak decline) 
 
Soluble mediators 
 
 
 
7.3.4. 
Safety Analysis 
Adverse events will be summarised by treatment and severity. Separate tables summarising 
adverse events and their relationship to study treatment as well as summarising adverse events 
by treatment and timepoint will also be produced.  
Serious adverse events will be listed and summarised by site, category and treatment. Separate 
tables will also be produced summarising serious adverse events and their relationship to study 
treatment as well as summarising serious adverse events by treatment and timepoint.  
Vital Signs and Laboratory blood tests will be summarized by treatment per visit and will be 
presented as means and standard deviations if data is consistent with that from a normal 
population distribution, and as medians and inter-quartile ranges for data that is inconsistent. 
Categorical variables will be presented as frequencies and percentages.  
If it is deemed appropriate, treatment effect will be assessed via t-test for continuous variables 
and Chi-square test or Fisher exact test for categorical variables (with the appropriate 
generalized linear model being used).  
 
 
 
 
 


ORCA_Asthma_SAP_v1.0 
 
page 17 of 35 
7.4. 
Exploratory Analysis 
7.4.1. 
Exploratory Mechanistic Testing 
Analysis to investigate any difference in effect of OC459 and placebo for the exploratory analyses 
below will be performed using the difference in values via unpaired (two-sample) t-test or Mann-
Whitney U tests for non-parametric data. 
 Changes in inflammatory cell counts by flow cytometry after drug treatment but before virus 
inoculation (for blood and nasal samples, not bronchial): Day -21 to Day -8. 
 Changes in soluble mediators after drug treatment but before virus inoculation (nasal samples): 
Day -21 to Day 0. 
 Changes in cell counts from bronchial biopsies taken at Day -8 and Day 5 will also be assessed 
CRTH2+ cells (cells/mm2 tissue) & EG2+ activated eosinophils (cells/mm2 tissue). 
 
7.4.2. 
Exploratory Testing For Antiviral Response 
Rhinovirus-induced IFN-β, λ1 and λ2/3 levels (protein and mRNA) will be measured in bronchial 
epithelial cell cultures stimulated with RV-16, RV-1B and Poly I:C after 6H, 24H and 48H. These will 
be compared across placebo and treatment groups, and to media and filtered virus stock controls. 
Data will be presented in a summary table and will include mean and standard error. Statistical 
analysis will be carried out for multiple comparisons using ANOVA, or where non-parametric 
methods are required, Wilcoxon or Kruskal-Wallis testing. Differences will be considered significant 
at P < 0.05. 
mRNA levels of a panel of interferon stimulated genes (MxA, RIG-I, OAS, viperin, IP-10, PKC) will also 
be analysed across the 3 timepoints using the same methods. 


ORCA_Asthma_SAP_v1.0 
 
page 18 of 35 
 
7.4.3. 
Exploratory Correlation Testing 
We will correlate lower respiratory symptom scores with the ACQ (which reflects symptoms over 
the last 7 days) at two timepoints: 
o ACQ-6 at day -9 vs sum of lower respiratory symptom scores day -15 to day -9 inclusive 
o ACQ-6 at day 10 vs sum of lower respiratory symptom scores day 4 to day 10 inclusive  
Potential relationships between the above will be investigated using Spearman’s rank correlations. 
Potential relationships between outcome variables and antiviral response variables will be 
investigated using Spearman’s rank correlations. Tests will be carried out between the following: 
 
Efficacy Analysis Variable 
 
Lower respiratory symptom score 
(peak and AUC) 
 
Upper respiratory symptom score 
(peak and AUC) 
 
Virus load (peak and AUC) 
 
Morning PEFR (peak decline) 
 
Morning FEV1 (peak decline) 
 
FeNO (peak increase) 
 
Any cytokine statistically induced by 
infection (peak increase)  
vs 
Antiviral Response Variable: 
 
interferon levels (β, λ1, λ2/3 ) 
 
mRNA levels of interferons (β, λ1, 
λ2/3)  
 
mRNA levels of a panel of interferon-
stimulated genes (MxA, RIG-I, OAS, 
viperin, IP-10, PKC) 
 
 
 
 
7.4.4. 
Further Testing For Outcome Variables 
In addition to the above exploratory tests, secondary outcome variables will be measured and 
analysed using alternative methods if deemed appropriate. Table 03 below contains a table 
showing the possible comparisons used: 


ORCA_Asthma_SAP_v1.0 
 
page 19 of 35 
Table 03: Potential Exploratory Analyses of Outcome Variables 
 
Comparison 
Variable: 
Peak during 
infection 
(day 0 to 10) 
Change from 
day 0 to peak 
infection 
Area under 
curve during 
infection 
(day 0 to 10) 
Total during 
infection (day 
0 to 14) 
Average 
value at each 
timepoint day 
0 to 10 
Lower 
respiratory 
symptom 
score1 
X 
X 
N/A 
X 
(primary 
outcome) 
X 
Upper 
respiratory 
symptom 
score1 
X 
X 
N/A 
X 
(secondary 
outcome) 
X 
Morning 
PEFR2 
X 
(peak fall) 
X 
(secondary 
outcome) 
X 
N/A 
X 
Morning 
FEV1
2 
X 
(peak fall) 
X 
(secondary 
outcome) 
X 
N/A 
X 
Clinic  
FEV1 2 
X 
(peak fall) 
X 
(secondary 
outcome) 
X 
N/A 
X 
ACQ-6 
(day 10) 
X 
(secondary 
outcome) 
N/A 
 
N/A 
N/A 
PC20 
(day 7) 
X 
(secondary 
outcome) 
N/A 
 
N/A 
(day -9 only) 
FeNO 
X 
X 
(secondary 
outcome) 
X 
 
N/A 
X 
Blood 
eosinophils 
X 
X 
(secondary 
outcome) 
N/A 
N/A 
X 
Virus load 
X 
(secondary 
outcome) 
N/A 
X 
 
N/A 
X 
Soluble 
mediators - 
nasal3 
X 
X 
(secondary 
mechanistic 
outcome) 
X 
N/A 
X 
Soluble 
mediators - 
bronchial3 
(day 5) 
X 
(secondary 
mechanistic 
outcome) 
N/A 
N/A 
(day -8 only) 
Items marked as ‘X’ or with a specific study day are to be assessed as exploratory analyses 


ORCA_Asthma_SAP_v1.0 
 
page 20 of 35 
7.5. 
Missing Data and Withdrawn Subjects 
Missing values for primary and secondary endpoints (as per Table 03), viral load and soluble 
mediators will be extrapolated for cumulative / area under the curve calculations.  
Imputation will be based upon the values before and after the missing timepoint(s). Missing 
values will be derived using the following formula: 
 
Where: 
 missing value at day ‘n’ 
 last recorded value before missing data begins 
 
is a constant derived from the last recorded value 
, the first value obtained after 
missing data , and   = the number of missing data points between 
 &  
  
 
For example; if we are missing data at day 4 and we know that the value at day 3 is 1 and the 
value at day 5 is 3 then we can calculate constant  as 
 thus the missing value at 
day 4 would be: 
.  
 
Imputation and analysis involving missing and imputed data will be taken under the assumption 
that the data is missing-at-random (MAR). 
If imputation has taken place, sensitivity analyses will be performed to ensure integrity of 
statistical testing based on extrapolated scores. 
No other values for missing data will be imputed. 
If there is a missing baseline value for virus load, the previous value (i.e. at randomisation / day -
21) will be taken (virus load should be below the lower limit of detection prior to inoculation). If 
there is a missing baseline value on another variable and after unblinding the subject is in the 
placebo group, the baseline will be replaced with a value from a previous visit (e.g. 
randomisation / day -21) if available. If after unblinding the subject is in the treatment group, the 
baseline will not be replaced (with the exception of virus load, as above). 
 


ORCA_Asthma_SAP_v1.0 
 
page 21 of 35 
 
7.6. 
Data Handling & Transformation 
ACQ scores will be calculated under the ACQ-6 variant of the form. The score will be derived by 
taking the mean average of the 6 values provided (note that whilst FEV is recorded this will not 
formulate part of the overall score).  
Viral loads below the limit of quantification will be treated as zero. Viral load data will be 
transformed using base 10 logarithm. To account for zero values, one will be added to each viral 
load measurement before being transformed. 
Soluble mediators which generate a detectable signal but are below the lower limit of detection 
will have an assumed value of half the lower limit of detection; those for which no signal was 
detected will have an assumed value of zero. Soluble mediators which are above the upper limit 
of detection will have an assumed value of the highest reading of all the samples on the same 
assay performed at that time. Normal transformation and/or sensitivity analyses may be 
performed as necessary depending on the data and the extent of values above and below limits 
of detection. 
AUC values for exploratory analyses will be calculated using the linear trapezoidal method. 
Missing values will be extrapolated where required. 
 
7.7. 
Software of Analysis 
Statistical analysis of primary and secondary endpoints will be performed using SAS 9.4. 
Additional graphical output may be performed using R. 
 


ORCA_Asthma_SAP_v1.0 
 
page 22 of 35 
 
APPENDIX A – INDEX OF TABLES, FIGURES & LISTINGS 
Baseline Variables 
Table 1.01  –  
Summary of Demographic Baseline Variables 
Table 1.02   –  
Summary of Demographic Variables at Randomisation (Day -21) 
Table 1.03   –  
Summary of Demographic Variables at Inoculation (Day 0) 
Primary Analysis 
Table 2.01   –  
Listing of Lower Respiratory System Scores 
Table 2.02   –  
Summary of Uncorrected Total Lower Respiratory System Scores 
Table 2.03   –  
Mann-Whitney U Test of Total Lower Respiratory System Scores 
Figure 2.04  –  
Box & Whisker Plot of Total Lower Respiratory System Scores 
Figure 2.05  –  
Box & Whisker Plot of Daily Lower Respiratory System Scores 
Secondary Efficacy Analysis 
Table 3.01   –  
Summary of Corrected Total Lower Respiratory System Scores 
Table 3.02   –  
Mann-Whitney U Test of Corrected Lower Respiratory System Scores 
Table 3.03   –  
Summary of Total Upper Respiratory System Scores 
Table 3.04   –  
Mann-Whitney U Test of Total Upper Respiratory System Scores 
Table 3.05   –  
Summary of Effect of Treatment on Asthma Exacerbation Variables 
Table 3.06   –  
Statistical Testing on Effect of Treatment on Asthma Exacerbation Variables 
Table 3.07   –  
Summary of Effect of Treatment on Viral Load Results 
Table 3.08   –  
Statistical Testing on Effect of Treatment Viral Load Results 
Secondary Mechanistic Analysis 
Table 3.09 -  
Summary of Nasal Soluble Mediators per visit 
Table 3.10 -  
Summary of Change from Baseline to Peak Value for Nasal Soluble Mediators  
Table 3.11 - 
Statistical Testing of Nasal Soluble Mediators 
Table 3.12 -  
Summary of Bronchial Soluble Mediators per visit (D-8, D5) 
Table 3.13 -  
Summary of Change from Baseline (D-8) to Infection (D5) for Bronchial Soluble 
Mediators  


ORCA_Asthma_SAP_v1.0 
 
page 23 of 35 
Table 3.14 - 
Statistical Testing of Bronchial Soluble Mediators 
Table 3.15 -  
Summary of Leukocyte Numbers per visit (D-8, D+5) 
Table 3.16 -  
Summary of Change from Baseline (D-8) to Infection (D+5) for Leukocyte Numbers 
Table 3.17 - 
Statistical Testing of Leukocyte Numbers 
Table 3.18 -  
Summary of Blood and Sputum Eosinophilia 
Table 3.19 -  
Summary of Change from Baseline to Peak Value for Blood and Sputum Eosinophilia 
Table 3.20 - 
Statistical Testing for Blood and Sputum Eosinophilia 
Secondary Analysis – Correlation Testing 
Table 3.21 - 
Spearmans Rank Correlation Testing 
Figure 3.22 - 
Correlation output 
Secondary Safety Analysis 
Table 3.23 - 
Summary of Adverse Events by Treatment and Timepoint 
Table 3.24 - 
Summary of Adverse Events by Treatment and Severity 
Table 3.25 - 
Summary of Adverse Events by Treatment and Relationship to Study Treatment 
Table 3.26 - 
Listing of Serious Adverse Events 
Table 3.27 - 
Summary of Serious Adverse Events by Treatment and Timepoint 
Table 3.28 - 
Summary of Serious Adverse Events by Treatment and Severity 
Table 3.29 - 
Summary of Serious Adverse Events by Treatment and Relationship to Study 
Treatment 
Table 3.30 - 
Summary of Vital Signs Results 
Table 3.31 - 
Summary of Clinical Chemistry & Haematology Results 
Exploratory Analysis 
Table 4.xx - 
Exploratory Analyses (as required) 
 
 


ORCA_Asthma_SAP_v1.0 
 
page 24 of 35 
APPENDIX B – SHELL TABLES, FIGURES & LISTINGS 
Baseline Variables 
Table 1.01  –  
Summary of Demographic Baseline Variables 
 Variable*  
Statistics 
OC459 
(N = xx) 
Placebo 
(N = xx) 
Age (y) 
N 
 
 
 
Mean 
 
 
 
SD 
 
 
 
Min 
 
 
 
Median 
 
 
 
Max 
 
 
 
*Variables for inclusion as per section 6.1 
 
Table 1.02   –  
Summary of Demographic Variables at Randomisation (Day -21) 
See Table 1.01 
Table 1.03   –  
Summary of Demographic Variables at Innoculation (Day 0) 
See Table 1.01 


ORCA_Asthma_SAP_v1.0 
 
page 25 of 35 
 
Primary Analysis 
Table 2.01   –  
Listing of Lower Respiratory System Scores 
 
Visit Scores 
 
Treatment 
Subject 
D0 
D1 
D2 
D3 
D4 
D5 
D6 
D7 
D8 
D9 D10 D11 D12 D13 D14 
Total 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Table 2.02   –  
Summary of Uncorrected Total Lower Respiratory System Scores 
Treatment 
Visit 
n 
Mean 
SD 
Median 
Min. 
Max. 
IQR 
OC459 
 
D0 
 
 
 
 
 
 
… 
 
 
 
 
 
 
D14 
 
 
 
 
 
 
Total 
 
 
 
 
 
 
Placebo 
 
D0 
 
 
 
 
 
 
… 
 
 
 
 
 
 
D14 
 
 
 
 
 
 
Total 
 
 
 
 
 
 
 


ORCA_Asthma_SAP_v1.0 
 
page 26 of 35 
 
Table 2.03   –  
Mann-Whitney U Test of Total Lower Respiratory System Scores 
Median Scores (Number of Points Above Median) for Variable Response 
Classified by Variable Treatment 
Treatment 
N 
Sum of 
Scores 
Expected 
Under H0 
Std Dev 
Under H0 
Mean 
Score 
Active 
xx 
xx 
xx 
xx 
xx 
Placebo 
xx 
xx 
xx 
xx 
xx 
Average scores were used for ties. 
 
Two-Sample Test 
Statistic (S) 
xx 
Normal Approximation 
 
Z 
xx 
One-Sided Pr > Z 
xx 
Two-Sided Pr > |Z| 
xx 
t Approximation 
 
One-Sided Pr > Z 
xx 
Two-Sided Pr > |Z| 
xx 
Exact Test 
 
One-Sided Pr >= S 
xx 
Two-Sided Pr >= |S - Mean| 
xx 
 
 
Figure 2.04  –  
Box & Whisker Plot of Total Lower Respiratory System Scores 
 


ORCA_Asthma_SAP_v1.0 
 
page 27 of 35 
 
Figure 2.05  –  
Box & Whisker Plot of Daily Lower Respiratory System Scores 
 
 
Secondary Efficacy Analysis 
Table 3.01   –  
Summary of Corrected Total Lower Respiratory System Scores 
See Table 2.02 
Table 3.02   –  
Mann-Whitney U Test of Corrected Lower Respiratory System Scores 
See Table 2.03 
Table 3.03   –  
Summary of Total Upper Respiratory System Scores 
See Table 2.02 
Table 3.04   –  
Mann-Whitney U Test of Total Upper Respiratory System Scores 
See Table 2.03 
Table 3.05   –  
Summary of Effect of Treatment on Asthma Exacerbation Variables 
Laboratory Test 
(units)* 
Treatment 
Visit 
n 
Mean 
SD 
Median 
Min. 
Max. 
IQR 
xxxxx (xxxx) 
Control 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Variables for inclusion as per section 6.3.1 


ORCA_Asthma_SAP_v1.0 
 
page 28 of 35 
Table 3.06   
–  
Statistical Testing on Effect of Treatment on Asthma Exacerbation Variables 
For normally distributed data variables: 
Source 
DF 
Sum of 
Squares 
Mean Square 
F Value 
Pf > F 
Model 
xx 
xx 
xx 
xx 
xx 
Error 
xx 
xx 
xx 
 
 
Corrected Total 
xx 
xx 
 
 
 
 
For non-parametric data variables: See Table 2.03 
 
Table 3.07   –  
Summary of Effect of Treatment on Viral Load Results 
Laboratory Test 
(units)* 
Treatment 
Visit 
n 
Mean 
SD 
Median 
Min. 
Max. 
IQR 
xxxxx (xxxx) 
Control 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Variables for inclusion as per section 6.3.1 
 
Table 3.08   –  
Statistical Testing on Effect of Treatment Viral Load Results 
For normally distributed data variables: 
Source 
DF 
Sum of 
Squares 
Mean Square 
F Value 
Pf > F 
Model 
xx 
xx 
xx 
xx 
xx 
Error 
xx 
xx 
xx 
 
 
Corrected Total 
xx 
xx 
 
 
 
 
For non-parametric data variables: See Table 2.03


ORCA_Asthma_SAP_v1.0 
 
page 29 of 35 
Secondary Mechanistic Analysis 
Table 3.09 -  
Summary of Nasal Soluble Mediators per visit 
Laboratory Test 
(units)* 
Treatment 
Visit 
n 
Mean 
SD 
Median 
Min. 
Max. 
IQR 
xxxxx (xxxx) 
Control 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Table 3.10 -  
Summary of Change from Baseline to Peak Value for Nasal Soluble Mediators  
See Table 3.09 
Table 3.11 - 
Statistical Testing of Nasal Soluble Mediators 
For normally distributed data variables: 
Source 
DF 
Sum of 
Squares 
Mean Square 
F Value 
Pf > F 
Model 
xx 
xx 
xx 
xx 
xx 
Error 
xx 
xx 
xx 
 
 
Corrected Total 
xx 
xx 
 
 
 
 
For non-parametric data variables: See Table 2.03 
 
Table 3.12 -  
Summary of Bronchial Soluble Mediators per visit (D-8, D5) 
See Table 3.09 
Table 3.13 -  
Summary of Change from Baseline (D-8) to Infection (D5) for Bronchial Soluble 
Mediators 
See Table 3.09 
Table 3.14 - 
Statistical Testing of Bronchial Soluble Mediators 
See Table 3.11 


ORCA_Asthma_SAP_v1.0 
 
page 30 of 35 
 
Table 3.15 -  
Summary of Leukocyte Numbers per visit (D-8, D5) 
See Table 3.09 
Table 3.16 -  
Summary of Change from Baseline (D-8) to Infection (D5) for Leukocyte Numbers 
See Table 3.09 
Table 3.17 - 
Statistical Testing of Leukocyte Numbers 
See Table 3.11  
 
Table 3.18 -  
Summary of Blood and Sputum Eosinophilia  
See Table 3.09 
Table 3.19 -  
Summary of Change from Baseline to Peak Value for Blood and Sputum Eosinophilia 
See Table 3.09 
Table 3.20 - 
Statistical Testing for Blood and Sputum Eosinophilia 
See Table 3.11 
 
 
 
Secondary Analysis – Correlation Testing 
Table 3.21 - 
Spearmans Rank Correlation Testing 
Pearson Correlation Coefficients 
Prob > |r| under H0: Rho=0  
Number of Observations 
  
Var 1
Var 2
Var 
xxxxxxx. 
xxxx
xxxx
xx
xxxx
xxxx
xx
Var 
xxxxxxx. 
xxxx
xxxx
xx
xxxx
xxxx
xx
 
 
 


ORCA_Asthma_SAP_v1.0 
 
page 31 of 35 
Figure 3.22 - 
Correlation Matrix  
 
The above matrix can contain the following: 
Baseline values of: 
 
ACQ (day 0) 
 
FeNO (day 0) 
 
Blood eosinophils (day -8) 
 
Serum IgE (day -8) 
 
PC20 (day -9) 
 
Skin prick test (from screening) 
o 
# of allergens 
o 
Sum of wheal diameters (mm) 
 
Soluble mediators 
 
vs 
Peak and AUC values of: 
 
Lower respiratory symptom score 
 
Upper respiratory symptom score 
 
Virus load 
 
Morning PEFR (peak decline) 
 
Morning FEV1 (peak decline) 
 
Soluble mediators 
 


ORCA_Asthma_SAP_v1.0 
 
page 32 of 35 
Secondary Safety Analysis 
Table 3.23 - 
Summary of Adverse Events by Treatment and Timepoint 
 
  
Events 
Patients 
 
OC459 
Placebo 
Total 
OC459 
Placebo 
Total 
 
Total Number of AEs 
 
 
 
 
 
 
Timepoint 
Pre-Treatment (Wk1) 
 
 
 
 
 
 
Pre-Inoculation (Wk2) 
 
 
 
 
 
 
Assessment Phase (Wk3-4) 
 
 
 
 
 
 
 
Follow-Up (Wk5-9) 
 
 
 
 
 
 
 
Table 3.24 - 
Summary of Adverse Events by Treatment and Severity 
 
  
Events 
Patients 
 
OC459 
Placebo 
Total 
OC459 
Placebo 
Total 
 
Number of 
Adverse events 
 
 
 
 
 
 
Severity 
Mild 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
Number of 
SAEs 
 
 
 
 
 
 
Reason for SAE 
Death 
 
 
 
 
 
 
Life threatening 
 
 
 
 
 
 
… 
 
 
 
 
 
 
Other medical 
important 
events 
 
 
 
 
 
 
 
Total 
 
 
 
 
 
 
 


ORCA_Asthma_SAP_v1.0 
 
page 33 of 35 
 
Table 3.25 - 
Summary of Adverse Events by Treatment and Relationship to Study Treatment 
 
All Events 
Treatment 
None 
Unlikely 
Possible 
Probable 
Definitely 
Not Yet 
Defined 
Total 
OC459 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
All subjects 
 
 
 
 
 
 
 
 
Patients with AEs* 
Treatment 
None 
Unlikely 
Possible 
Probable 
Definitely 
Not Yet 
Defined 
Total 
OC459 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
All subjects 
 
 
 
 
 
 
 
*Table Note: Where subjects have more than one AE the highest relationship has been used. 
 
Table 3.26 - 
Listing of Serious Adverse Events 
OC459: 
Subj. 
AE 
Diagnosis 
Details 
Relation to 
Treatment 
Start Date 
Ongoing 
/End Date Duration 
Expected-
ness 
Severity 
 
 
 
 
Placebo: 
Subj. 
AE 
Diagnosis 
Details 
Relation to 
Treatment 
Start Date 
Ongoing 
/End Date Duration 
Expected-
ness 
Severity 
 
 
 
 
 
Table 3.27 - 
Summary of Serious Adverse Events by Treatment and Timepoint 
 
  
Events 
Patients 
 
OC459 
Placebo 
Total 
OC459 
Placebo 
Total 
 
Total Number of SAEs 
 
 
 
 
 
 
Timepoint 
Pre-Treatment (Wk1) 
 
 
 
 
 
 
Pre-Inoculation (Wk2) 
 
 
 
 
 
 
Assessment Phase (Wk3-4) 
 
 
 
 
 
 
 
Follow-Up (Wk5-9) 
 
 
 
 
 
 
 


ORCA_Asthma_SAP_v1.0 
 
page 34 of 35 
 
Table 3.28 - 
Summary of Serious Adverse Events by Treatment and Severity 
 
  
Events 
Patients 
 
OC459 
Placebo 
Total 
OC459 
Placebo 
Total 
 
Number of 
Serious 
Adverse events 
 
 
 
 
 
 
Severity 
Mild 
 
 
 
 
 
 
Moderate 
 
 
 
 
 
 
Severe 
 
 
 
 
 
 
 
Table 3.29 - 
Summary of Serious Adverse Events by Treatment and Relationship to Study 
Treatment 
 
All Serious Adverse Events 
Treatment 
None 
Unlikely 
Possible 
Probable 
Definitely 
Not Yet 
Defined 
Total 
OC459 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
All subjects 
 
 
 
 
 
 
 
 
Patients with SAEs* 
Treatment 
None 
Unlikely 
Possible 
Probable 
Definitely 
Not Yet 
Defined 
Total 
OC459 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
All subjects 
 
 
 
 
 
 
 
*Table Note: Where subjects have more than one SAE the highest relationship has been used. 
 


ORCA_Asthma_SAP_v1.0 
 
page 35 of 35 
 
Table 3.30 - 
Summary of Vital Signs Results 
Laboratory Test 
(units)* 
Treatment 
Visit 
n 
Mean 
SD 
Median 
Min. 
Max. 
xxxxx (xxxx) 
Control 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Variables for inclusion as per section 6.3.4 
 
Table 3.31 - 
Summary of Clinical Chemistry & Haematology Results 
Laboratory Test 
(units)* 
Treatment 
Visit 
n 
Mean 
SD 
Median 
Min. 
Max. 
xxxxx (xxxx) 
Control 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Variables for inclusion as per section 6.3.4 
 
Exploratory Analysis 
Table 4.xx - 
Exploratory Analyses as required 
Extent of output determined by Section 7.4