Conditions: Idiopathic Pulmonary Fibrosis
Keywords: TRK-250 Idiopathic Pulmonary Fibrosis
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 34
Healthy volunteers: no

Brief summary:
TRK-250 is a nucleic acid medicine that inhibits the progression of pulmonary fibrosis by selectively suppressing the expression of transforming growth factor-beta 1 (TGF-β1) protein, at the gene expression level. This study is a double-blind, randomized, placebo-controlled Phase I study. The primary objective of the study is to assess the safety and tolerability of single and multiple inhaled doses of TRK-250 in subjects with idiopathic pulmonary fibrosis (IPF).

Detailed description:

Arms:
arm 1: None arm 2: None

Interventions: intervention 1: TRK-250 intervention 2: Placebo
Intervention descriptions: intervention 1: single and multiple doses (4 weeks) intervention 2: single and multiple doses (4 weeks)

Protocol document:
Prot_000.pdf: 
TITLE PAGE 
Protocol 
TRK-250 – A Phase I, Double-Blind, Placebo-Controlled, Single and 
Multiple Inhaled Dose, Safety, Tolerability, and Pharmacokinetic Study of 
TRK-250 in Subjects with Idiopathic Pulmonary Fibrosis 
Protocol Amendment Status: Final 
Original Protocol Date: 01 June 2018 
Protocol Date: 24 September 2018 
Protocol Version: Amendment 3 
Investigational Product: TRK-250 
Sponsor Reference Number: 250IPF01 
 
PIND Number: 133065 
Sponsor: 
Toray Industries, Inc. 
1-1, Nihonbashi-muromachi 2-chome, 
Chuo-ku, Tokyo 
Tokyo, 103-8666 Japan 
 
Emergency Medical Contact 
US: 
 
(this number is not to be used for submitting 
serious adverse event or pregnancy reports; 
please see Appendix 1) 
Information described herein is confidential and may be disclosed only with the express 
written permission of the Sponsor. 
Page 11 of 75


Protocol Amendment 3 
 
CONFIDENTIAL 
Sponsor Reference: 250IPFO 1 
SPONSOR APPROVAL 
I have read the protocol and approve it: 
 
 
 
Final Protocol Amendment 3, 24 September 2018 
Page 2 of65 
Page 12 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 3 of 65 
INVESTIGATOR AGREEMENT 
I have read the protocol and agree to conduct the study as described herein. 
<Name, qualification(s)> 
<Institution> 
Date 
<Position in organization> 
Page 13 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 4 of 65 
STUDY IDENTIFICATION 
Sponsor 
Toray Industries, Inc. 
1-1, Nihonbashi-muromachi 2-chome 
Chuo-ku, Tokyo, 103-8666 Japan 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 14 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 5 of 65 
SYNOPSIS 
Title of study: TRK-250 – A Phase I, Double-Blind, Placebo-Controlled, Single and Multiple 
Inhaled Dose, Safety, Tolerability, and Pharmacokinetic Study of TRK-250 in Subjects with 
Idiopathic Pulmonary Fibrosis 
Objectives:  
The primary objective of the study is to assess the safety and tolerability of single and multiple 
inhaled doses of TRK-250 in subjects with idiopathic pulmonary fibrosis (IPF). 
The secondary objective of the study is to assess the pharmacokinetics (PK) in blood following 
single and multiple inhaled doses of TRK-250 in subjects with IPF. 
 
Endpoints: 
The primary safety endpoints for this study are as follows: 
•
body weight
•
vital signs measurements
•
oxygen saturation (SpO2) by pulse oximetry
•
hematology, clinical chemistry, and urinalysis test results
•
12-lead electrocardiogram (ECG) parameters
•
forced expiratory volume over 1 second (FEV1), forced vital capacity (FVC)
•
incidence and severity of adverse events (AE)
The single and multiple ascending dose PK outcome endpoints of TRK-250 are as follows (where 
applicable): 
•
area under the concentration-time curve (AUC) from time zero to the time of the last
quantifiable  concentration (AUC0-tlast)
•
maximum observed concentration (Cmax)
•
time to Cmax (Tmax)
Other PK parameters may also be added. 
 
Page 15 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 6 of 65 
Study design: 
This will be a Phase I, double-blind, randomized, placebo-controlled study conducted in 2 parts, 
single and multiple inhaled dose, at multiple sites. 
Part A 
Part A will comprise a single-dose, single-period, sequential-cohort study. Potential subjects will be 
screened to assess their eligibility to enter the study within 28 days prior to study drug 
administration. 
It is planned that 16 subjects will be studied in 4 cohorts (Cohorts A1 to A4), each consisting of 
4 subjects. Following data monitoring committee (DMC) review of the safety and tolerability data 
from the first 4 cohorts, up to 3 further optional cohorts (Cohorts A5 to A7) of at least 4 subjects 
may be added to Part A to evaluate additional doses or to further evaluate planned doses. 
Investigational treatment and follow-up duration: 
The first 2 subjects to be dosed in each cohort of Part A will reside at the study site from Day 1 (the 
day of dosing) to Day 2 (24 hours post completion of dose administration). After Investigator 
review of the safety data through Day 2 from the first 2 subjects, the remaining subjects in 
Cohort A1 may be discharged from the study site after completion of all study procedures on Day 1, 
returning for subsequent procedures on an outpatient basis. For subsequent cohorts, the decision of 
when to discharge subjects from the study site will be made by the Investigator based on the safety 
data through Day 2 from the first 2 subjects of the current cohort, as well as any relevant 
recommendations made at the previous DMC dose escalation review meeting.  
All subjects will return for a Safety Follow-up visit 7 (+3) days after dosing and an Additional 
Follow-up visit 14 (±3) days after dosing. The timing of the Additional Follow-up visit for a cohort 
may be changed to a maximum of 28 (±3) days after dosing, based on emerging safety data. 
 
 
 
 
Dose levels: 
Current proposed dose levels: 
•
Cohort A1:  2 mg
•
Cohort A2:  10 mg
•
Cohort A3:  30 mg
•
Cohort A4:  60 mg
 
  
Page 16 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 7 of 65 
Part B 
Part B will comprise a multiple-dose, single-period, sequential-cohort study. Potential subjects will 
be screened to assess their eligibility to enter the study within 28 days prior to initial study drug 
administration. 
It is planned to study 18 subjects in 3 cohorts (Cohorts B1 to B3), each consisting of 6 subjects. Up 
to 3 further optional cohorts (Cohorts B4 to B6) of at least 6 subjects may be added to evaluate 
additional doses or to further evaluate planned doses. 
Investigational treatment and follow-up duration: 
After Screening, subjects will be dosed at the study site on Days 1, 8, 15, and 22. Subjects will 
return for a Safety Follow-up visit approximately 7 days after their last dose (Day 29 [+3]) and an 
Additional Follow-up visit approximately 14 days after the last dose (Day 36 [±3]). The timing of 
the Additional Follow-up visit for a cohort may be changed to a maximum of approximately 
28 days after the last dose (Day 50 [±3]), based on emerging safety data. 
 
 
 
 
 
 
  
 
 
 
  
Dose levels: 
To be confirmed following completion of Part A. Current proposed dose levels: 
•
Cohort B1: 10 mg
•
Cohort B2: 30 mg
•
Cohort B3: 60 mg
 
Number of subjects: 
Part A: 16 subjects will be studied in 4 groups (Groups A1 to A4), with the option to add up to 
3 additional cohorts of at least 4 subjects at DMC recommendation. 
Part B: 18 subjects will be studied in 3 groups (Groups B1 to B3), with the option to add up to 
3 additional cohorts of at least 6 subjects at DMC recommendation. 
Diagnosis and main criteria for inclusion: 
Male and female subjects aged between 40 and 80 years (inclusive) with IPF. Subjects must have 
SpO2 ≥90% at rest by pulse oximetry while breathing ambient air, FVC and FEV1 ≥50% of 
predicted, ratio of FEV1 to FVC ≥0.7, and DLCO corrected for hemoglobin 30% to 79% of 
predicted, inclusive. 
Investigational products, dose, and mode of administration: 
Test product: 
TRK-250 solution, formulated for inhalation via InnoSpire Go nebulizer 
system (Philips) 
Proposed dose levels for Part A: 2, 10, 30, and 60 mg 
Proposed dose levels for Part B: 10, 30, and 60 mg once weekly for 4 weeks. The dosing frequency 
and dosing duration for Part B will be decided on the basis of data from Part A of the study. The 
daily dose administered will not exceed the highest dose administered in Part A. 
Administration route: inhalation 
Page 17 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 8 of 65 
Reference product and mode of administration: 
Reference product: placebo solution, formulated for inhalation via InnoSpire Go nebulizer system 
(Philips) 
Administration route: inhalation 
Duration of subject participation in the study: 
Planned Screening duration: approximately 28 days. 
Planned study duration (Screening to Follow-up):  
Part A: planned to be no more than 7 weeks, but may be increased to a maximum of 9 weeks if the 
Additional Follow-up visit is delayed based on findings from the ongoing review of data by the 
DMC. 
Part B: planned to be no more than 10 weeks, but may be increased to a maximum of 12 weeks if 
the Additional Follow-up visit is delayed based on findings from the ongoing review of data by the 
DMC. 
Analysis methods: 
Pharmacokinetics: 
Blood samples for the analysis of TRK-250 concentrations. Pharmacokinetic parameters will be 
derived by noncompartmental analysis. For both study parts, the PK parameters will include 
AUC0-tlast, Cmax, and Tmax. Other PK parameters may also be added. 
 
Safety: 
AE, pulse oximetry, spirometry, DLCO, body weight, clinical laboratory evaluations (hematology, 
clinical chemistry, urinalysis, serology), 12-lead electrocardiograms, vital sign measurements, and 
physical examinations. 
Statistical methods: 
Pharmacokinetics: 
Individual blood concentrations of TRK-250 and PK parameters will be listed and summarized 
using descriptive statistics. 
 
 
Safety: 
Safety parameters will be listed and summarized using descriptive statistics. No inferential 
statistical analysis of safety data is planned. 
Page 18 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 9 of 65 
TABLE OF CONTENTS 
TITLE PAGE .............................................................................................................................1 
SPONSOR APPROVAL ...........................................................................................................2 
INVESTIGATOR AGREEMENT.............................................................................................3 
STUDY IDENTIFICATION .....................................................................................................4 
SYNOPSIS .................................................................................................................................5 
TABLE OF CONTENTS ...........................................................................................................9 
LIST OF TABLES AND FIGURES........................................................................................11 
LIST OF ABBREVIATIONS ..................................................................................................12 
1. 
INTRODUCTION ...........................................................................................................14 
1.1. 
Overview ................................................................................................................14 
 
 
 
 
 
 
 
 
1.6. 
Summary of Clinical Experience ...........................................................................18 
1.7. 
Study Rationale ......................................................................................................18 
1.8. 
Benefit-Risk Assessment .......................................................................................19 
2. 
OBJECTIVES AND ENDPOINTS .................................................................................19 
2.1. 
Objectives ..............................................................................................................19 
2.2. 
Endpoints ...............................................................................................................19 
2.2.1. 
Primary Endpoints .......................................................................................19 
2.2.2. 
Secondary Endpoints ...................................................................................20 
 
 
3. 
INVESTIGATIONAL PLAN ..........................................................................................20 
3.1. 
Overall Study Design and Plan ..............................................................................20 
3.1.1. 
Part A ...........................................................................................................20 
3.1.2. 
Part B ...........................................................................................................22 
3.2. 
Study Start and End of Study Definitions ..............................................................24 
3.3. 
Additional Cohorts .................................................................................................24 
3.4. 
Discussion of Study Design, Including the Choice of Control Groups .................24 
3.4.1. 
Dose Interval ................................................................................................25 
3.5. 
Selection of Doses in the Study .............................................................................26 
 
 
 
 
 
 
 
 
 
 
4. 
SELECTION OF STUDY POPULATION .....................................................................31 
Page 19 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 10 of 65 
4.1. 
Inclusion Criteria ...................................................................................................31 
4.2. 
Exclusion Criteria ..................................................................................................32 
4.3. 
Subject Number and Identification ........................................................................34 
4.4. 
Subject Withdrawal and Replacement ...................................................................34 
4.5. 
Study Termination .................................................................................................34 
5. 
STUDY TREATMENTS .................................................................................................35 
5.1. 
Description, Storage, Packaging, and Labeling .....................................................35 
5.2. 
Study Treatment Administration............................................................................35 
5.2.1. 
Secondary Exposure Protection Measures ...................................................36 
5.3. 
Randomization .......................................................................................................36 
5.4. 
Blinding..................................................................................................................36 
5.5. 
Treatment Compliance ...........................................................................................37 
5.6. 
Drug Accountability...............................................................................................37 
6. 
CONCOMITANT THERAPIES AND OTHER RESTRICTIONS ................................37 
6.1. 
Concomitant Therapies ..........................................................................................37 
6.1.1. 
Restricted Concomitant Therapies ...............................................................37 
6.1.2. 
Prohibited Concomitant Therapies ..............................................................38 
6.2. 
Diet .........................................................................................................................38 
6.3. 
Smoking .................................................................................................................38 
6.4. 
Blood Donation ......................................................................................................38 
6.5. 
Contraception .........................................................................................................38 
7. 
STUDY ASSESSMENTS AND PROCEDURES ...........................................................39 
7.1. 
Safety and Tolerability Assessments .....................................................................40 
7.1.1. 
Adverse Events ............................................................................................40 
7.1.2. 
Clinical Laboratory Evaluations ..................................................................40 
7.1.3. 
Vital Signs ....................................................................................................41 
7.1.4. 
12-Lead Electrocardiogram .........................................................................41 
7.1.5. 
Physical Examination...................................................................................41 
7.1.6. 
Body Weight and Height .............................................................................42 
7.1.7. 
Pulse Oximetry.............................................................................................42 
7.1.8. 
Spirometry....................................................................................................42 
7.1.9. 
Carbon Monoxide Diffusion Capacity Test .................................................44 
7.1.10. 
Diagnosis of IPF ..........................................................................................45 
7.1.10.1. 
High-Resolution Computed Tomography..........................................45 
7.1.10.2. 
Surgical Lung Biopsy ........................................................................45 
7.1.10.3. 
Combined Eligibility Criteria ............................................................45 
7.1.11. 
Emergency Sponsor Contact ........................................................................46 
7.2. 
Pharmacokinetic Assessments ...............................................................................46 
7.2.1. 
Pharmacokinetic Sample Collection and Processing ...................................46 
7.2.2. 
Pharmacokinetic Analytical Methodology ..................................................46 
Page 20 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 11 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
8. 
SAMPLE SIZE AND DATA ANALYSIS ......................................................................48 
8.1. 
Determination of Sample Size ...............................................................................48 
8.2. 
Analysis Populations ..............................................................................................48 
8.2.1. 
Safety Population .........................................................................................48 
8.2.2. 
Pharmacokinetic Population ........................................................................49 
 
 
8.3. 
Safety Analysis ......................................................................................................49 
8.4. 
Pharmacokinetic Analyses .....................................................................................49 
 
 
8.6. 
Interim Analysis .....................................................................................................49 
9. 
REFERENCES ................................................................................................................49 
10. APPENDICES .................................................................................................................51 
Appendix 1: Adverse Event Reporting ..............................................................................52 
Appendix 2: Clinical Laboratory Evaluations ...................................................................56 
Appendix 3: Total Blood Volume......................................................................................57 
Appendix 4: Regulatory, Ethical, and Study Oversight Considerations ............................58 
Appendix 5: Schedule of Assessments ..............................................................................61 
 
LIST OF TABLES AND FIGURES 
Table 1: 
Toxicokinetic Parameters in Rat Lung Following a Single Dose of 
TRK-250 Inhaled over 60 Minutes (LC/MS-HRAM) .................................................17 
Table 2: 
Toxicokinetic Parameters in Rat Lung Following a Single Dose of 
TRK-250 Inhaled over 60 Minutes (LC-FD) ...............................................................18 
Table 3: 
Concentrations of TRK-250 (and Drug-Associated Fluorescence) in 
Cynomolgus Monkey Lung Tissues Post Dosing on Day 28 Following 4 Weeks 
Repeated Dosing (n = 3) (LC-FD) ...............................................................................18 
Table 4: 
Proposed IMP Dose Levels for Parts A and B ...........................................27 
Table 5: 
Preparation of TRK-250 Doses for Nebulization ......................................35 
Table 6:  
Diagnosis of IPF ........................................................................................46 
 
Figure 1: 
Study Schematic (Part A)...........................................................................22 
Figure 2: 
Study Schematic (Part B) ...........................................................................24 
 
Page 21 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 12 of 65 
 
 
LIST OF ABBREVIATIONS 
Abbreviation 
Definition 
ABCA3 
adenosine triphosphate binding cassette A3 
AE 
adverse event(s) 
ALAT 
Latin American Thoracic Association 
ALT 
alanine aminotransferase 
AST 
aspartate aminotransferase 
ATS 
American Thoracic Society 
AUC 
area under the concentration-time curve 
AUC0-24h 
area under the concentration-time curve from time 0 to 24 hours 
AUC0-tlast 
area under the concentration-time curve from time zero to the time of the 
last quantifiable concentration 
 
 
 
 
CFR 
Code of Federal Regulations 
CIOMS 
Council for International Organizations of Medical Sciences 
Cmax 
maximum observed concentration 
CRF 
Case Report Form(s) 
CRO 
Contract Research Organization 
CSA 
clinical study agreement 
DLCO 
carbon monoxide diffusion capacity 
DMC 
data monitoring committee 
ECG 
electrocardiogram 
EDC 
electronic data capture 
ERS 
European Respiratory Society 
FDA 
Food and Drug Administration 
FEV1 
forced expiratory volume over 1 second 
FLP 
full length product 
FVC 
forced vital capacity 
FVC%pred 
percent of predicted forced vital capacity 
GCP 
Good Clinical Practice 
GLI 
Global Lung Initiative 
GLP 
Good Laboratory Practice 
HEPA 
high efficiency particulate air 
HRCT 
high-resolution computed tomography 
IB 
Investigator’s Brochure 
IC50 
concentration that reduces effect by 50% 
ICF 
Informed Consent Form 
Page 22 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 13 of 65 
ICH 
International Council for Harmonisation 
IMP 
investigational medicinal product 
IPF 
idiopathic pulmonary fibrosis 
IRB 
Institutional Review Board 
IUD 
intrauterine device 
IxRS 
interactive voice/web response system 
JRS 
Japanese Respiratory Society 
LC-FD 
liquid chromatography with fluorescence detection 
LC/MS-HRAM 
liquid chromatography/mass spectrometry-high resolution accurate mass 
LLOQ 
lower limit of quantification 
MABEL 
minimum anticipated biological effect level 
MedDRA 
Medical Dictionary for Regulatory Activities 
 
 
 
 
 
 
mRNA 
messenger ribonucleic acid 
 
 
NOAEL 
no-observed-adverse-effect level 
 
 
PK 
pharmacokinetic(s) 
QTcB 
QT interval corrected for heart rate using Bazett’s method 
QTcF 
QT interval corrected for heart rate using Fridericia’s method 
SAE 
serious adverse event 
siRNA 
small, interfering ribonucleic acid 
SLB 
surgical lung biopsy(ies) 
SOP 
standard operating procedure(s) 
SP-A2 
surfactant protein A2 
SP-C 
surfactant protein C 
SpO2 
oxygen saturation 
TGF-β1 
transforming growth factor-beta 1 
Tmax 
time of maximum observed concentration 
TMF 
Trial Master File 
UIP 
usual interstitial pneumonia 
ULN 
upper limit of normal 
WCBP 
women of childbearing potential 
 
 
Page 23 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 14 of 65 
1. INTRODUCTION 
1.1. 
Overview 
Interstitial pulmonary fibrosis (IPF) is one of the interstitial pneumonias, known also as 
cryptogenic fibrosing alveolitis. The condition is also identified as usual interstitial 
pneumonia (UIP), the term given its appearance on high-resolution computed tomography 
(HRCT) scans of the lung and histopathologic examination of surgical lung biopsies (SLB).1 
Toray Industries, Inc. (Toray), is developing TRK-250 for the potential treatment of IPF. 
An international, professional society (American Thoracic Society/European Respiratory 
Society/Japanese Respiratory Society/Latin American Thoracic Association 
[ATS/ERS/JRS/ALAT]) statement defines IPF as a specific lung disorder of adults, of 
unknown cause, characterized by progressive dyspnea, associated with poor prognosis, 
resulting from progressive fibrosing interstitial pneumonia of unknown cause.2 Criteria for 
the diagnosis of IPF detailed in this publication are adopted as criteria central to establishing 
subject eligibility for this clinical trial. 
The etiology of IPF remains undetermined. Risk factors have been identified, including: 
• cigarette smoking and atmospheric pollutants 
• viral infection 
• chronic aspiration 
• drugs 
Genetic predisposition seems to play a role in some cases of IPF.3 Mutations in surfactant 
proteins A2 (SP-A2) and C (SP-C) as well as in adenosine triphosphate binding cassette A3 
(ABCA3) genes appear to be involved in cases of familial IPF. A genetic abnormality in the 
promoter of the gene encoding mucin 5B (MUC5B) has been identified in about one third of 
individuals with familial and sporadic IPF; the abnormality is only found in about 9% of 
individuals without clinical evidence of IPF. Telomerase gene mutations (TERT, TERC, 
DKC1, TINF2) that cause shortened telomeres have been identified in about one quarter of 
sporadic and more than one third of familial IPF cases. Still, the interplay of all risk factors 
remains poorly understood, more so in sporadic IPF cases. 
A central role for transforming growth factor-beta 1 (TGF-β1) has been documented for 
decades in fibrosis in many organs, in airway remodeling, and in IPF.4 TRK-250 is a small, 
interfering ribonucleic acid (siRNA) molecule that targets the TGF-β1 guide sequence. 
TRK-250 aims to inhibit the progression of pulmonary fibrosis and, thus, slow the decline in 
lung function, by selectively targeting TGF-β1 messenger ribonucleic acid (mRNA) and 
thereby suppressing the expression of TGF-β1 protein, a key growth factor involved in lung 
fibrosis, at the gene expression level.5 TRK-250 employs a proprietary nucleic acid platform 
that synthesizes 2 complementary single oligoribonucleotide strands. The 2 strands of 
oligoribonucleotides are linked by a proline derivative and a second proline derivative 
terminated with guanine completes the sequence. TRK-250 is formulated as a solution for 
inhalation, supporting direct administration to the lung. 
Page 24 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 15 of 65 
This trial will be the first time that TRK-250 is administered to humans. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 25 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 16 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
Page 26 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 17 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 27 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 18 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
1.6. 
Summary of Clinical Experience 
TRK-250 has not previously been administered to humans. 
1.7. 
Study Rationale 
The principal aim of this study is to obtain safety and tolerability data when TRK-250 is 
administered by inhalation as single and multiple doses to subjects with IPF. A 
sequential-cohort, ascending-dose design has been chosen to optimize safety since Part A of 
the study will be the first time TRK-250 is administered to humans.  
Conducting the study in subjects with IPF was considered necessary due to the paucity of 
data concerning administration of siRNA, in general, and TRK-250, specifically, to healthy 
subjects; a perception shared by regulatory authorities. 
Inhaled administration has been chosen for the study since this is the intended route of 
clinical administration. 
PK blood sampling is included in this trial design. This should advance understanding of the 
potential for inhaled TRK-250 to have off-target effects. 
Once-weekly dosing frequency has been selected for Part B because 4 once-weekly 
intratracheal doses of TRK-250 were shown to be pharmacologically active in a human 
TGF-β1 
 model.  
Page 28 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 19 of 65 
Based on the nonclinical data, the duration of treatment in each part of this trial is considered 
adequate to achieve the study’s objectives. 
This study will be double blind and placebo controlled to avoid bias in the collection and 
evaluation of data. Placebo has been chosen as the control treatment to assess whether any 
observed effects are treatment related or simply reflect the study conditions. 
Ongoing review of safety and tolerability data will be performed before deciding to proceed 
to the next dose escalation.
 
 
1.8. 
Benefit-Risk Assessment 
Subjects with IPF in the current study are not expected to gain significant, long-term health 
benefits (beyond that of an assessment of their medical status) from participating in the study. 
The risks of participation are primarily those associated with adverse reactions to the 
investigational medicinal product (IMP), although there may also be some discomfort from 
collection of blood samples and other study procedures.  
Although there is no evidence from nonclinical studies of bronchoconstriction following 
inhalation of TRK-250, subjects will have spirometry performed to assess for this important 
safety finding in this trial. 
2. OBJECTIVES AND ENDPOINTS 
2.1. 
Objectives 
The primary objective of the study is to assess the safety and tolerability of single and 
multiple inhaled doses of TRK-250 in subjects with IPF. 
The secondary objective of the study is to assess the PK in blood following single and 
multiple inhaled doses of TRK-250 in subjects with IPF.  
 
 
 
 
 
2.2. 
Endpoints  
2.2.1. 
Primary Endpoints 
The primary safety endpoints for this study are as follows: 
• body weight 
• vital signs measurements 
Page 29 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 20 of 65 
• oxygen saturation (SpO2) by pulse oximetry  
• hematology, clinical chemistry, and urinalysis test results 
• 12-lead electrocardiogram (ECG) parameters 
• forced expiratory volume over 1 second (FEV1), forced vital capacity (FVC) 
• incidence and severity of adverse events (AE) 
2.2.2. 
Secondary Endpoints 
The single and multiple ascending dose PK outcome endpoints of TRK-250 are as follows 
(where applicable): 
• AUC from time zero to the time of the last quantifiable concentration (AUC0-tlast) 
• Cmax 
• Tmax 
Other PK parameters may also be added. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
3. INVESTIGATIONAL PLAN 
This will be a double-blind, randomized, placebo-controlled study conducted in 2 parts, 
single and multiple inhaled dose, at multiple sites. 
3.1. 
Overall Study Design and Plan 
3.1.1. 
Part A 
Part A will comprise a single-dose, single-period, sequential-cohort study. Potential subjects 
will be screened to assess their eligibility to enter the study within 28 days prior to study drug 
administration. 
The total duration of study participation for each subject (from Screening through Additional 
Follow-up visit) is planned to be no more than 7 weeks but may be increased to a maximum 
Page 30 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 21 of 65 
of 9 weeks if the Additional Follow-up visit is delayed based on findings from the ongoing 
review of data by the data monitoring committee (DMC). 
It is planned that 16 subjects will be studied in 4 cohorts (Cohorts A1 to A4), each consisting 
of 4 subjects. Following DMC review of the safety and tolerability data from the first 
4 cohorts, up to 3 further optional cohorts (Cohorts A5 to A7) of at least 4 subjects may be 
added to Part A to evaluate additional doses or to further evaluate planned doses. 
 
 
 
 
 
 
 
 
 
 
 
  
All subjects will return for a Safety Follow-up visit 7 (+3) days after dosing and an 
Additional Follow-up visit 14 (±3) days after dosing. The timing of the Additional Follow-up 
visit for a cohort may be changed to a maximum of 28 (±3) days after dosing, based on 
emerging safety data. 
In each of Cohorts A1 to A4, 3 subjects will receive TRK-250 and 1 subject will receive 
placebo. All doses will be administered in accordance with a randomization schedule on the 
morning of Day 1. Each subject will receive only a single dose of TRK-250 or placebo during 
the study. 
 
 
 
 
 
 
 
• Cohort A1: 2 mg 
• Cohort A2: 10 mg 
• Cohort A3: 30 mg 
• Cohort A4: 60 mg 
Page 31 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 22 of 65 
 
 
 
 
 
 
 
 pharmacologically active dose levels (ie, when 
the escalated dose will be ≥34.7 mg).  
An overview of the study design and the planned dose levels are shown in Figure 1. 
Figure 1: 
Study Schematic (Part A) 
 
3.1.2. 
Part B 
Part B will comprise a multiple-dose, single-period, sequential-cohort study. Potential 
subjects will be screened to assess their eligibility to enter the study within 28 days prior to 
initial study drug administration. 
The total duration of study participation for each subject (from Screening through Safety 
Follow-up visit) is planned to be no more than 10 weeks but may be increased to a maximum 
of 12 weeks if the Additional Follow-up visit is delayed based on findings from the ongoing 
review of data by the DMC. 
 
 
Page 32 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 23 of 65 
 
  
 
  
 
  
All subjects will return for a Safety Follow-up visit approximately 7 days after their last dose 
(Day 29 [+3]) and an Additional Follow-up visit approximately 14 days after the last dose 
(Day 36 [±3]). The timing of the Additional Follow-up visit for a cohort may be changed to a 
maximum of approximately 28 days after the last dose (Day 50 [±3]), based on emerging 
safety data. 
In each of Cohorts B1 to B3, 4 subjects will receive TRK-250 and 2 subjects will receive 
placebo. Doses will be administered on the morning of Days 1, 8, 15, and 22 in accordance 
with a randomization schedule. Each subject will receive 4 once-weekly doses of TRK-250 or 
placebo during the study. 
 
 
 
 
 
 
• Cohort B1: 10 mg 
• Cohort B2: 30 mg 
• Cohort B3: 60 mg 
 
 
 
 
  
shown in Figure 2. 
Page 33 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 24 of 65 
Figure 2: 
Study Schematic (Part B) 
  
 
A Schedule of Assessments is presented in Appendix 5. 
3.2. 
 Study Start and End of Study Definitions 
The start of the study is defined as the date the first enrolled subject signs an Informed 
Consent Form (ICF). The point of randomization occurs at the time of subject number 
allocation. The end of the study is defined as the date of the last subject’s last 
protocol-specified assessment, whether scheduled or unscheduled. 
3.3. 
Additional Cohorts 
Following review of the safety and tolerability data from the planned dose cohorts, the DMC 
may recommend additional dose cohorts to be added to the study. Up to 3 further cohorts of 
at least 4 subjects
 
 may be included in Part A, and up to 3 further cohorts of at least 6 subjects 
 
 may be 
included in Part B. If the dose escalation stopping criteria have not been met, the doses 
administered in the additional cohorts may exceed the proposed highest doses shown in 
Table 4;
 
 
The Sponsor is responsible for making the final decision whether to add any additional 
cohorts if recommended by the DMC. The decision will be documented in the Trial Master 
File (TMF), and the Institutional Review Board (IRB) will be notified of the changes. 
3.4. 
Discussion of Study Design, Including the Choice of Control Groups 
For both parts of the study, a sequential-cohort, ascending-dose design has been chosen for 
safety reasons because TRK-250 is in the early stages of clinical development, with Part A of 
Page 34 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 25 of 65 
the study being the first time it will be administered to humans. Inhaled doses have been 
chosen for both parts of the study since this is the intended clinical route of administration.  
 
 
  
 
 
This study will be double blind and placebo controlled to avoid bias in the collection and 
evaluation of data. Placebo has been chosen as the control treatment to assess whether any 
observed effects are treatment related or simply reflect the study conditions. 
 
 
 
 
 
3.4.1. 
Dose Interval 
 
 
 
 
 
 
 
 
 
 
 
 
• 
 
 
 
Page 35 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 26 of 65 
 
 
 
 
 
3.5. 
Selection of Doses in the Study 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
Page 36 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 27 of 65 
 
 
 
 
 
  
 
 
 
  
The proposed IMP dose levels for Parts A and B are shown in Table 4. 
Table 4: 
Proposed IMP Dose Levels for Parts A and B 
Study Part 
Cohort 
Treatment  
Part A 
A1 
2 mg TRK-250 or placebo 
 
A2 
10 mg TRK-250 or placebo 
 
A3 
30 mg TRK-250 or placebo 
 
A4 
60 mg TRK-250 or placebo 
 
Optional Cohorts A5, A6, and A7 
XX mg TRK-250 or placebo 
Part B 
B1 
10 mg TRK-250 or placebo Q1W 
 
B2 
30 mg TRK-250 or placebo Q1W 
 
B3 
60 mg TRK-250 or placebo Q1W 
 
Optional Cohorts B4, B5, and B6 
XX mg TRK-250 or placebo Q1W 
Abbreviations: IMP = investigational medicinal product; Q1W = once weekly. 
For Part B of the study, dose levels, dosing frequency, and dosing duration will be decided, in 
consultation with the Sponsor, on the basis of data from Part A of the study. The weekly dose 
of TRK-250 administered during this part of the study will not exceed the maximum 
well-tolerated dose level studied in Part A. 
 
 
 
Details of all doses administered in Parts A and B of the study will be documented in the 
TMF. 
 
 
 
 
Page 37 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 28 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
Page 38 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 29 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 39 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 30 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
Page 40 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 31 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
4. SELECTION OF STUDY POPULATION 
4.1. 
Inclusion Criteria 
Subjects must satisfy all of the following criteria, at the Screening visit unless otherwise 
stated: 
1. Able to comprehend and willing to sign an ICF and to abide by the study restrictions. 
2. Male or female between 40 and 80 years of age, inclusive, at Screening. 
a. Women of childbearing potential (WCBP) and all male subjects will agree to 
use contraception (refer to Section 6.5) throughout enrollment (ie, from 
signing the ICF to the last protocol-specified assessment, whether scheduled 
or unscheduled). 
3. Clinical, radiographic, and (when available) histologic features consistent with IPF 
within 5 years prior to Screening, confirmed using ATS/ERS/JRS/ALAT statement2 
for the diagnosis of IPF in effect at the time of this trial’s initiation. 
Page 41 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 32 of 65 
a. Interpretation of HRCT and, if performed, SLB must be confirmed by central 
over-reader (see Section 7.1.10.1 and Section 7.1.10.2). 
b. HRCT performed within 12 months prior to the initiation of Screening that is 
determined to be of suitable quality by the central over-reader will be used to 
confirm subject eligibility. 
4. SpO2 ≥90% at rest by pulse oximetry while breathing ambient air. 
5. FVC ≥50% of predicted. 
6. FEV1 ≥50% of predicted. 
7. Ratio of FEV1 to FVC ≥0.7. 
8. DLCO corrected for hemoglobin 30% to 79% of predicted, inclusive. 
9. Life expectancy of at least 12 months. 
4.2. 
Exclusion Criteria 
Subjects will be excluded from the study if they satisfy any of the following criteria on or 
before Day 1, as applicable: 
1. Serious or uncontrolled medical, surgical, or psychiatric disease that in the opinion of 
the Investigator would compromise subject safety.  
2. History of acute exacerbation of IPF or respiratory tract infection within 3 months 
prior to Screening. 
3. Planned surgery during the study (Day 1 to Additional Follow-up Visit). 
4. History of malignant tumor within 5 years prior to Screening (with the exception of 
treated squamous and basal cell skin cancers and treated Stage 0/in situ cervical 
cancer). 
5. History of emphysema or clinically significant respiratory diseases (other than IPF), 
as determined by the Investigator. 
6. History of significant hypersensitivity, intolerance, or allergy to any drug compound, 
food, or other substance, unless approved by the Investigator. 
7. Pregnant or planning to become pregnant during enrollment. 
8. Breastfeeding or planning to breastfeed during enrollment. 
9. History of alcoholism or drug/chemical abuse within 2 years prior to Day 1. 
10. Alcohol consumption >21 units per week for males and >14 units per week for 
females within 30 days prior to Screening. One unit of alcohol equals 12 oz (360 mL) 
beer, 1½ oz (45 mL) liquor, or 5 oz (150 mL) wine. Eligible subjects must have 
agreed not to exceed this limit until the end of the subject’s enrollment. 
11. Positive urine drugs of abuse at Screening for substance or metabolite of substance 
that was not prescribed and that remains positive on repeat testing.  
12. Positive hepatitis panel (positive for either hepatitis B surface antigen, hepatitis B 
core antibody, or hepatitis C antibody) and/or positive human immunodeficiency 
Page 42 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 33 of 65 
virus test. Subjects whose results are compatible with prior immunization and not 
infection may be included at the discretion of the Investigator. 
13. Participation in a clinical study involving administration of an investigational drug 
within 30 days or 5 half-lives, whichever is longer, prior to Day 1.  
14. Use or intend to use any medications/products known to alter TRK-250 absorption, 
metabolism, or elimination processes within 30 days prior to Day 1 and for the 
remainder of enrollment, unless deemed acceptable by the Investigator. 
15. Use or intend to use any nonprescription medications/products including vitamins, 
minerals, and phytotherapeutic/herbal/plant-derived preparations within 7 days prior 
to Day 1 and for the remainder of enrollment, unless deemed acceptable by the 
Investigator. 
16. Consumption in any form of tobacco- or nicotine-containing products, excluding 
over-the-counter or prescription nicotine replacement products approved for 
marketing by the FDA, within 3 months prior to Day 1.  
17. Receipt of blood products within 3 months prior to Day 1. 
18. Donation of blood, plasma, or platelets within 6 weeks prior to Day 1.  
19. Unable to perform spirometry or to inhale study drug properly, in the opinion of the 
Investigator. In particular, subjects whose cough may impede proper study treatment, 
in the opinion of the Investigator, should be excluded. 
20. Poor peripheral venous access. 
21. Have previously received TRK-250. 
22. A subject who, in the opinion of the Investigator, should not participate in this study. 
23. Findings that are diagnostic of an alternative condition other than UIP on surgical 
lung biopsy, HRCT imaging, transbronchial lung biopsy, or BAL. 
24. Other known causes of interstitial lung disease (eg, drug toxicities, environmental 
exposures, connective tissue diseases). 
25. End-stage fibrotic disease expected to require organ transplantation within 6 months. 
26. Taking a systemic corticosteroid, cytotoxic therapy (eg, chlorambucil, azathioprine, 
cyclophosphamide, or methotrexate), vasodilator therapy for pulmonary hypertension 
(eg, bosentan), or unapproved treatment for IPF (eg, interferon-gamma, penicillamine, 
cyclosporine, mycophenolate, or N-acetylcysteine) within 4 weeks prior to Screening.  
a. Treatment with pirfenidone (Esbriet®) or nintedanib (Ofev®), though not both 
concurrently, is permitted, provided that the subject has been on a stable dose for 
at least 4 weeks prior to Screening and it is anticipated the dose will remain 
unchanged throughout enrollment (ie, from signing the ICF to the last 
protocol-specified assessment, whether scheduled or unscheduled). 
27. Has received an investigational therapy within 5 half-lives of the agent or 6 months 
prior to Screening, whichever is longer. 
Page 43 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 34 of 65 
4.3. 
Subject Number and Identification 
Subjects will be assigned a subject number prior to the first dosing occasion at the time of 
their randomization. Assignment of subject numbers will be in ascending order. 
Subjects will be identified by subject number only on all study documentation. A list 
identifying the subjects by subject number will be kept in the Site Master File. 
4.4. 
Subject Withdrawal and Replacement 
A subject is free to withdraw from the study at any time. In addition, a subject will be 
withdrawn if any of the following criteria are met: 
• change in compliance with any inclusion/exclusion criterion post randomization that 
is clinically relevant and affects subject safety as determined by the Investigator. 
• noncompliance with the study restrictions that might affect subject safety or study 
assessments/objectives, as considered applicable by the Investigator. 
• any clinically relevant sign or symptom that, in the opinion of the Investigator, 
warrants subject withdrawal. 
If a subject is withdrawn, the Sponsor and 
will be notified by the Investigator and 
the date and reason(s) for the withdrawal will be documented in the subject’s Case Report 
Form (CRF). If a subject is withdrawn, efforts will be made to perform all follow-up 
assessments, including the Additional Follow-up visit assessments, if possible (Appendix 5). 
Other procedures may be performed at the Investigator’s or Sponsor’s discretion. If the 
subject is in-house, the Safety Follow-up procedures should be performed before the subject 
is discharged from the clinic, if possible. The Investigator may also request that the subject 
return for a separate follow-up visit to complete the Safety Follow-up procedures. Any 
withdrawn subject who completes the Safety Follow-up procedures should be invited to 
attend for a separate follow-up visit to complete the procedures scheduled for the Additional 
Follow-up visit; this additional visit should be performed approximately 1 week after the 
Safety Follow-up procedures. All withdrawn subjects will be followed until resolution of all 
their AE or until the unresolved AE are judged by the Investigator to have stabilized. 
Subjects who are withdrawn for reasons not related to study drug, including randomized 
subjects who never receive study drug, may be replaced following discussion between the 
Investigator and the Sponsor to ensure the required number of subjects complete each cohort. 
Subjects withdrawn as a result of an AE thought to be related to the study drug will generally 
not be replaced. 
4.5. 
Study Termination 
The study may be discontinued at the discretion of the Investigator or Sponsor if any of the 
following criteria are met: 
• medical or ethical reasons affecting the continued performance of the study 
• difficulties in the recruitment of subjects 
• cancellation of drug development 
Page 44 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 35 of 65 
• the DMC recommends discontinuation of the study due to safety concerns. 
5. STUDY TREATMENTS 
5.1. 
Description, Storage, Packaging, and Labeling 
The IMP
 will be supplied by 
the Sponsor (or designee), along with the batch/lot numbers and Certificates of Analysis. The 
IMP will be provided as a clear and sterile solution, formulated for inhalation via InnoSpire 
Go nebulizer system (Philips). 
 
 
 
The bulk drug container and unit dose containers will be labeled in accordance with national 
laws and regulations and packaged together with the diluent as kit boxes. Study treatment 
(IMP or placebo) will be prepared for administration by qualified site personnel in 
accordance with Table 5. 
Table 5: 
Preparation of TRK-250 Doses for Nebulization 
Dose 
2 mg 
(or Placebo) 
10 mg 
(or Placebo) 
30 mg 
(or Placebo) 
60 mg 
(or Placebo) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
5.2. 
Study Treatment Administration 
Each dose of TRK-250 and placebo will be administered by oral inhalation using InnoSpire 
Go. Measures to protect against secondary exposure will be employed as described in 
Section 5.2.1. 
Study treatment must be administered by blinded site personnel. 
Subjects will be administered the study treatment in a sitting position. All doses will be 
administered irrespective of fasting status. 
Subjects must be instructed not to disclose to anyone whether the study treatment is 
associated with a taste or odor. 
Prior to administering study drug, the Investigator should passively observe the subject’s 
cough character and frequency to establish a “qualitative” baseline. The subject should be 
asked to rate informally her/his shortness of breath (eg, on a scale of 0 to 10 or none, mild, 
moderate, or severe). 
After starting study drug administration and for roughly 30 minutes afterwards, the 
Investigator should continue to passively observe the subject for change in character and/or 
Page 45 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 36 of 65 
frequency of cough. Special attention should be given to assessing whether the subject’s 
cough may impede proper study treatment. 
The same informal self-assessment should be performed for chest tightness. A substantial 
fraction of patients experience no change in cough or chest tightness during methacholine or 
histamine challenge even with ≥20% decline in FEV1, but a change in cough and/or chest 
tightness are good and early indicators of worsening bronchoconstriction. 
5.2.1. 
Secondary Exposure Protection Measures 
Study treatment may be administered in a negative pressure enclosure or in proximity to a 
portable high efficiency particulate air (HEPA) filtration system intended for healthcare in 
accordance with local standard operating procedures (SOP) and/or study documents. 
 
 
 
 
 
 
5.3. 
Randomization 
Randomization to placebo/active will be performed using an interactive voice/web response 
system (IxRS) randomization service. Investigators must register the subject for the IxRS 
system in sequence of the screening completion and dispense study drug to each subject per 
the code allocated by IxRS. This unique drug code will not be changed throughout the study. 
5.4. 
Blinding 
The following controls will be employed to maintain the double-blind status of the study: 
• The placebo solution will be identical in appearance to TRK-250. 
• The Investigator and other site personnel not involved in the preparation of study 
treatment will remain blinded to study treatment randomization. 
• Respiratory protection must be used (as described in Section 5.2.1) during study 
treatment administration to prevent site personnel from detecting any taste or odor of 
the study treatment. 
• Subjects must be instructed not to disclose to anyone whether study treatment is 
associated with a taste or odor. 
• Interim bioanalytical data will be provided in a blinded manner. 
Page 46 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 37 of 65 
The IxRS will be programmed with blind-breaking instructions. The study blind may be 
broken if, in the opinion of the Investigator, it is in the subject’s best interest to know the 
study treatment assignment. Whenever possible, and providing it does not interfere with or 
delay any decision in the best interest of the subject, the Investigator will discuss the intended 
code-break with the Sponsor. If it becomes necessary to break the code during the study, the 
date, time, and reason will be recorded in the subject’s source data and CRF, as appropriate. 
5.5. 
Treatment Compliance 
To ensure treatment compliance, all doses of study treatment will be administered under the 
supervision of suitably qualified site personnel. 
5.6. 
Drug Accountability 
The Investigator will maintain an accurate record of receipt of study supplies. In addition, an 
accurate drug disposition record will be kept, specifying the amount dispensed to each subject 
and the date of dispensing. This drug accountability record will be available for inspection at 
any time. At the completion of the study, the original drug accountability record will be 
available for review by 
or the Sponsor on request. 
For each batch of unit doses, the empty used unit dose containers will be discarded upon 
satisfactory completion of the compliance and accountability procedures. Any unused 
assembled unit doses will be retained until completion of the study. 
At the completion of the study, all unused supplies will be returned to 
(or designee) 
or may be destroyed at the study site in accordance with 
 or the Sponsor’s written 
instructions and local SOP. 
6. CONCOMITANT THERAPIES AND OTHER RESTRICTIONS 
6.1. 
Concomitant Therapies 
6.1.1. 
Restricted Concomitant Therapies 
Subjects should be instructed to withhold for at least 4 hours before planned study visits any 
short-acting bronchodilator therapy (beta-agonist or anticholinergic agents). Withheld doses 
may be taken after the spirometry session scheduled to occur within 30 minutes after 
completion of study treatment administration.  
Each subject will refrain from use of any new prescription or nonprescription medications 
during enrollment until the last planned or unscheduled study visit, unless clinically indicated 
to safeguard the subject’s safety or well-being.  
Either pirfenidone or nintedanib must have been taken at a stable dose for at least 4 weeks 
prior to Screening and should be continued at the same dose throughout a subject’s 
enrollment (ie, from signing the ICF to the last protocol-specified assessment, whether 
scheduled or unscheduled). Dose adjustment in accordance with the prescribing information 
is permitted if clinically indicated. 
Page 47 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 38 of 65 
All other therapies are permitted concomitant therapies. All prescription medications should 
be continued at the same dose throughout a subject’s enrollment (ie, from signing the ICF to 
the last protocol-specified assessment, whether scheduled or unscheduled), unless a change is 
clinically indicated to safeguard the subject’s safety or well-being. Consistent compliance and 
dosing of inhaled therapies is particularly important. 
Any medication taken by a subject during the course of the study and the reason for its use 
will be documented in the source data and in the relevant CRF. 
6.1.2. 
Prohibited Concomitant Therapies 
Taking pirfenidone and nintedanib simultaneously is prohibited for subjects enrolled in this 
study. 
Taking a systemic corticosteroid, cytotoxic therapy (eg, chlorambucil, azathioprine, 
cyclophosphamide, or methotrexate), vasodilator therapy for pulmonary hypertension (eg, 
bosentan), unapproved treatment for IPF (eg, interferon-gamma, penicillamine, cyclosporine, 
mycophenolate, or N-acetylcysteine), or any other investigational treatment is prohibited for 
subjects enrolled in this study. 
6.2. 
Diet 
While confined at the study site, subjects will receive a standardized diet at scheduled times 
that do not conflict with other study-related activities. Subjects will be fasted overnight (at 
least 8 hours) before collection of blood samples for safety laboratory tests.  
On the days with PK assessments (Day 1 for Part A and Days 1 and 22 for Part B), meals will 
be identical for each cohort. 
Water will be freely available at all times. 
Consumption of alcohol >21 units per week for males and >14 units per week for females is 
not permitted within 30 days prior to Screening until the end of the subject’s enrollment. One 
unit of alcohol equals 12 oz (360 mL) beer, 1½ oz (45 mL) liquor, or 5 oz (150 mL) wine. 
6.3. 
Smoking 
Subjects may not consume in any form tobacco- or nicotine-containing products, excluding 
over-the-counter or prescription nicotine replacement products approved for marketing by the 
FDA, within 3 months prior to Day 1 until the end of the subject’s enrollment. 
6.4. 
Blood Donation 
Subjects must not donate blood, plasma, or platelets from 6 weeks prior to Day 1 until 
56 days after the end of the subject’s enrollment.  
6.5. 
Contraception 
Women who are not of childbearing potential will not be required to use contraception. 
Women not of childbearing potential are defined as: 
Page 48 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 39 of 65 
• permanently sterile due to hysterectomy or bilateral oophorectomy confirmed by 
history, or  
• postmenopausal, defined as at least 12 months postcessation of menses without an 
alternative medical cause, confirmed by a Screening blood follicle-stimulating 
hormone level >40 mIU/mL.  
Subjects who are WCBP must be willing to use a highly effective method of birth control (ie, 
contraceptive measure with a failure rate of <1% per year) in conjunction with male barrier 
contraception (ie, male condom with spermicide) from the time of signing the ICF until 
90 days after the last dose of study treatment. The highly effective method of contraception 
should be started at least 1 menstrual cycle prior to the first dose of study drug. Highly 
effective methods of contraception are: 
• intrauterine device (IUD; eg, Mirena®). Steel or copper IUDs are not acceptable.  
• established use of oral, implantable, transdermal, or injectable hormonal method of 
contraception associated with inhibition of ovulation.  
• male sterilization (performed at least 90 days prior to the Screening visit), with verbal 
confirmation of surgical success (for female subjects on the study, the vasectomized 
male partner should be the sole partner for that subject). 
• bilateral salpingectomy, tubal ligation, or tubal occlusion (performed at least 90 days 
prior to the Screening visit). 
Male subjects will be surgically sterile for at least 90 days prior to Screening or will be 
required to use a male condom with spermicide from Day 1 until 90 days after the last dose of 
study drug when sexually active with female partners of childbearing potential. Sexual 
intercourse with female partners who are pregnant or breastfeeding should be avoided unless 
condoms are used from the time of the first dose until 90 days after the last dose of study 
drug. Male subjects are required to refrain from donation of sperm from Day 1 until 90 days 
after the last dose of study drug. 
Subjects who practice true abstinence, because of the subject’s lifestyle choice (ie, the subject 
should not become abstinent just for the purpose of study participation), are exempt from 
contraceptive requirements. Periodic abstinence (eg, calendar, ovulation, symptothermal, 
postovulation methods) and coital withdrawal are not acceptable methods of contraception. If 
a subject who is abstinent at the time of signing the ICF becomes sexually active she/he must 
agree to use contraception as described previously.  
For subjects who are exclusively in same-sex relationships, contraceptive requirements do not 
apply. If a subject who is in a same-sex relationship at the time of signing the ICF becomes 
heterosexually active, they must agree to use contraception as described above.  
7. STUDY ASSESSMENTS AND PROCEDURES 
Every effort will be made to schedule and perform the procedures as closely as possible to the 
nominal time, giving considerations to appropriate posture conditions, practical restrictions, 
and the other procedures to be performed at the same timepoint. 
Page 49 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 40 of 65 
The order of priority for scheduling procedures around a timepoint is (in descending order of 
priority): 
• dosing 
• blood sampling 
• spirometry 
• any other procedures 
 (ECGs will be scheduled before vital 
signs measurements). 
Spirometry should preferably be performed before DLCO at timepoints where both 
assessments are required (but see Section 7.1.8). 
 
 
7.1. 
Safety and Tolerability Assessments 
7.1.1. 
Adverse Events 
AE definitions, assignment of seriousness, severity, and causality, and procedures for 
reporting SAE are detailed in Appendix 1. 
The condition of each subject will be monitored from the time of signing the ICF to final 
discharge from the study. Subjects will be observed for any signs or symptoms and asked 
about their condition by open questioning, such as “How have you been feeling since you 
were last asked?”, at least once each day while resident at the study site and at each study 
visit. Subjects will also be encouraged to spontaneously report AE occurring at any other 
time during the study. 
All nonserious AE, whether reported by the subject voluntarily or upon questioning, or noted 
on physical examination, will be recorded from initiation of study drug until study 
completion. SAE will be recorded from the time the subject signs the ICF until study 
completion. The nature, time of onset, duration, seriousness, and severity will be 
documented, together with an Investigator’s opinion of the relationship to study drug. 
SAE will be followed up until the subject’s status reverts to baseline, the AE is determined by 
the Investigator to be irreversible, or death of the subject. Non-serious AE will be followed 
up until resolution or the AE is determined by the Investigator to be irreversible. This will be 
completed at the Investigator’s discretion. If not possible to follow up an SAE or AE, 
appropriate reasons should be provided. 
7.1.2. 
Clinical Laboratory Evaluations 
Blood and urine samples will be collected for clinical laboratory evaluations (including 
clinical chemistry, hematology, urinalysis, and serology) at the times indicated in the 
Schedule of Assessments in Appendix 5. Clinical laboratory evaluations are listed in 
Appendix 2. Additional clinical laboratory evaluations will be performed at other times if 
judged to be clinically appropriate or if the ongoing review of the data suggests a more 
Page 50 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 41 of 65 
detailed assessment of clinical laboratory safety evaluations is required. Maximum blood 
volumes that will be collected in this study for clinical laboratory evaluations and other blood 
collections are listed in Appendix 3. 
Subjects will be asked to provide a urine sample for drugs of abuse screen at Screening. For 
all WCBP, a pregnancy test will be performed at the times indicated in the Schedule of 
Assessments in Appendix 5.  
An Investigator will perform a clinical assessment of all Screening clinical laboratory data. 
7.1.3. 
Vital Signs 
Blood pressure, pulse rate, respiratory rate, and oral body temperature will be assessed with 
the subject sitting at rest for at least 5 minutes at the times indicated in the Schedule of 
Assessments in Appendix 5. Vital signs may also be performed at other times if judged to be 
clinically appropriate or if the ongoing review of the data suggests a more detailed 
assessment of vital signs is required. 
Day 1 predose blood pressure, pulse rate, and respiratory rate will be measured in triplicate at 
approximately 2-minute intervals. The median value will be used as the baseline value in the 
data analysis. All subsequent measurements will be performed singly and repeated once if 
outside the relevant clinical reference ranges. Oral body temperature will be measured singly. 
7.1.4. 
12-Lead Electrocardiogram 
Resting 12-lead ECGs will be recorded after the subject has been supine and at rest for at 
least 5 minutes at the times indicated in the Schedule of Assessments in Appendix 5. Single 
12-lead ECGs will be repeated once if either of the following criteria apply: 
• QTcF value >500 ms 
• QTcF change from the baseline (Day 1 predose) is >60 ms. 
If repeated, the repeat values will be used for data analysis. 
Additional 12-lead ECGs may be performed at other times if judged to be clinically 
appropriate or if the ongoing review of the data suggests a more detailed assessment of ECGs 
is required. The Investigator will perform a clinical assessment of each 12-lead ECG.  
The ECG machine will compute the PR and QT intervals, QTc, QRS duration, and heart rate. 
The QT interval corrected for heart rate using Bazett’s method (QTcB) and QTcF will also be 
calculated. A common set of reference ranges will be applied to all the preceding ECG 
parameters at all sites. 
7.1.5. 
Physical Examination 
A full physical examination or symptom-directed physical examination will be performed at 
the timepoints specified in the Schedule of Assessments in Appendix 5.  
Full physical examination will include the following body systems: general appearance, 
eyes/ears/nose/throat/head/neck, chest and lungs (including inspection of the thorax for scars 
Page 51 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 42 of 65 
consistent with SLB at Screening), cardiovascular, abdomen, musculoskeletal, lymphatic, 
dermatologic, neurologic, psychiatric, and extremities. 
7.1.6. 
Body Weight and Height 
Body weight (in underclothes) and height will be recorded at the times indicated in the 
Schedule of Assessments in Appendix 5. 
7.1.7. 
Pulse Oximetry 
Pulse oximetry will be performed with the subject sitting at rest for at least 5 minutes at the 
times indicated in the Schedule of Assessments in Appendix 5. 
The Screening assessment will be conducted while the subject is breathing ambient air. 
If the subject’s subsequent status requires supplemental oxygen by nasal prongs/cannula, the 
initial SpO2 measurement should be obtained with the subject breathing the currently 
prescribed oxygen supplementation. Using clinical judgement to assure the subject’s safety 
and well-being, an attempt should be made to obtain SpO2 with the subject breathing oxygen 
at 2 L/minute by nasal prongs/cannula for 5 minutes (less time if desaturation is rapid and 
clinically significant). Then, an attempt should be made to obtain SpO2 with the subject 
breathing ambient air for 5 minutes, again using clinical judgement to assure the subject’s 
safety and well-being. 
7.1.8. 
Spirometry 
Spirometry testing will be conducted to measure FVC, FEV1, and FEV1/FVC at the times 
indicated in the Schedule of Assessments in Appendix 5.  
Spirometry will be performed and interpreted in accordance with published guidelines.9,10 
Spirometry predicted values will be calculated using Global Lung Initiative (GLI) 2012 
equations.11 
At any study visit when performance of both spirometry and DLCO are planned, the preferred 
order is for spirometry to be performed before DLCO. If this should not be feasible, the 
rationale must be well documented in source records. Whatever order of testing is 
implemented at Day 1 is the order of testing to be used at every subsequent study visit. 
A subject may only proceed to Day 1 when Screening spirometry is determined to be 
adequate by the central over-reader. If Screening spirometry is determined to be inadequate 
by the central over-reader, the subject must return for repeat testing at the earliest feasible 
date. If the subject reaches the end of the Screening window (see Appendix 5) without 
documentation of adequate spirometry as determined by the central over-reader, the subject 
will fail Screening and may rescreen once. 
Baseline spirometry values are those obtained at Day 1. In the event that Day 1 spirometry is 
determined not to be adequate by the central over-reader and the subject has received study 
treatment, baseline spirometry values are those obtained at Screening. 
Page 52 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 43 of 65 
When planning repeat spirometry, the turnaround time required for central over-read (see the 
vendor’s study manual) should be taken into consideration. When repeating either spirometry 
or DLCO determined to be inadequate by the central over-reader, only the inadequate modality 
(spirometry and/or DLCO) needs to be repeated. 
Spirometry may only be performed on subjects in this study by site personnel certified as 
competent to perform the assessments. Whenever possible, all spirometry on a single subject 
should be performed: 
• at the same time of day (±2 hours) as for the initial session (ie, Screening) 
• on the same equipment 
• by the same site personnel 
• under the same conditions, including the order in which spirometry and DLCO 
assessments at the same timepoint are performed 
Spirometry tests will be performed in accordance with bronchodilator restrictions specified in 
Section 6.1.1. 
At study visits when study treatment is to be administered, the following schedule of 
spirometry assessments (forced expiratory maneuvers) is recommended: 
• a minimum of 3, maximum of 6 maneuvers within 60 minutes predose 
• preferably 3 but a minimum of 2 maneuvers as soon as feasible but within 15 minutes 
after completion of dose administration 
• preferably 3 but a minimum of 2 maneuvers at 30 ± 5 minutes after completion of 
dose administration 
• preferably 3 but a minimum of 2 maneuvers at 60 ± 10 minutes after completion of 
dose administration 
• preferably 3 but a minimum of 2 maneuvers at 90 ± 15 minutes after completion of 
dose administration 
The maximum number of forced expiratory maneuvers in any day should not exceed 16. The 
minimum number of maneuvers at study visits when study treatment is to be administered 
should be 11 but it is acknowledged this may be too taxing for an individual with IPF. The 
Investigator should use clinical judgement to balance the safe monitoring of the subject for 
possible bronchoconstriction following study treatment against the burden of this testing. 
Greater emphasis should be placed on proper assessment closer to the time when treatment is 
completed than at later times. In the absence of evidence of bronchoconstriction and if the 
subject is not able to perform the schedule of assessments proposed above, the Investigator 
may alter the timing of postdose spirometry sessions, ie, spread them out over a longer 
interval, or may reduce the number of postdose spirometry sessions. The Investigator may not 
alter the specified predose spirometry assessment. At every postdose spirometry session, a 
minimum of 2 forced expiratory maneuvers must be attempted. 
If the Investigator implements changes to the proposed postdose spirometry assessments, the 
rationale must be documented in detail in source records. 
Page 53 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 44 of 65 
If the Investigator determines it is necessary to alter postdose spirometry, the decision applies 
only to the current visit. At all subsequent study visits, efforts to implement the proposed 
schedule of assessments should be renewed. 
Clinical judgement is also foreseen to predicate how to perform assessments when postdose 
bronchoconstriction is suspected. In this circumstance, the number and timing of forced 
expiratory maneuvers is to be determined by the Investigator. Ideally, there should be a final 
assessment demonstrating resolution of bronchoconstriction, ie, FEV1 and FVC decline 
<12% from predose values. 
Detailed spirometry testing instructions will be provided to sites in a separate document. 
7.1.9. 
Carbon Monoxide Diffusion Capacity Test 
In order to assess pulmonary gas exchange in subjects, testing will be conducted to measure 
DLCO at the times indicated in the Schedule of Assessments in Appendix 5. 
DLCO will be performed and interpreted in accordance with published guidelines.12 DLCO 
predicted values will be calculated using GLI 2017 equations.13 
A subject may only proceed to Day 1 when Screening DLCO is determined to be adequate by 
the central over-reader. If Screening DLCO is determined to be inadequate by the central 
over-reader, the subject must return for repeat testing at the earliest feasible date. If the 
subject reaches the end of the Screening window (see Appendix 5) without documentation of 
adequate DLCO as determined by the central over-reader, the subject will fail Screening and 
may rescreen once. 
Baseline DLCO values are those obtained at Day 1. In the event that Day 1 DLCO is 
determined not to be adequate by the central over-reader and the subject has received study 
treatment, baseline DLCO values are those obtained at Screening. 
When planning repeat DLCO, the turnaround time required for central over-read (see the 
vendor’s study manual) should be taken into consideration. When repeating either spirometry 
or DLCO determined to be inadequate by the central over-reader, only the inadequate modality 
(spirometry and/or DLCO) needs to be repeated. 
Testing for DLCO may only be performed on subjects in this study by site personnel certified 
as competent to perform the assessments. Whenever possible, all DLCO on a single subject 
should be performed: 
• at the same time of day (±2 hours) as for the initial session (ie, Screening) 
• on the same equipment 
• by the same site personnel 
• under the same conditions, including the order in which spirometry and DLCO 
assessments at the same timepoint are performed 
DLCO will be performed in accordance with bronchodilator restrictions specified in 
Section 6.1.1. 
Page 54 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 45 of 65 
DLCO will be corrected for hemoglobin. Hemoglobin may be measured locally within 7 days 
before the DLCO measurement. Alternatively, the hemoglobin value reported by the central 
laboratory obtained at the study visit during which DLCO was performed may be used. 
Detailed DLCO testing instructions will be provided to sites in a separate document. 
7.1.10. 
Diagnosis of IPF 
The diagnosis of IPF will be confirmed using ATS/ERS/JRS/ALAT consensus criteria.2 The 
consensus criteria are modified with regard to HRCT confirmation of UIP as follows. 
7.1.10.1. 
High-Resolution Computed Tomography 
HRCT assessment will be conducted for Screening purposes only, as part of confirming the 
diagnosis of UIP/IPF (see Schedule of Assessments in Appendix 5). HRCT must qualify 
based on Table 4 in ATS/ERS/JRS/ALAT statement for the diagnosis of IPF.2 
If the most recent historical HRCT was performed more than 12 months prior to the initiation 
of Screening, OR if the HRCT performed within 12 months prior to the initiation of 
Screening is determined not of suitable quality by the central over-reader, the imaging study 
must be repeated during Screening. Any HRCT used to establish subject eligibility must 
undergo central interpretation. Details of the central over-read vendor will be provided to 
study sites in a separate document. 
7.1.10.2. 
Surgical Lung Biopsy 
Site personnel are expected to confirm at Screening whether the subject has undergone SLB 
by: detailed review of the medical record; focused questioning when obtaining the medical 
history; and physical examination focused on detection of scars suggestive of SLB. 
If SLB has been performed, efforts to obtain these slides must be initiated immediately. Once 
the slides are in the site’s possession, they must be handled per the central over-reader’s 
instructions with special attention to obscuring all identifying information. Then, the slides 
will be submitted to the independent, central over-read vendor. Details of the central 
over-read vendor will be provided to study sites in a separate document. 
SLB will be interpreted on the basis of Table 5 in Raghu et al. 2011.2 For the purposes of this 
trial, the column labeled Possible UIP Pattern will be considered synonymous with 
Nonclassifiable Fibrosis as discussed in the ATS/ERS/JRS/ALAT statement’s text under UIP 
Pattern: Histopathology Features; see Section 7.1.10.3. 
7.1.10.3. 
Combined Eligibility Criteria 
Table 6 adds the preceding refinements to Table 6 in the ATS/ERS/JRS/ALAT statement.2 
Page 55 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 46 of 65 
Table 6:  
Diagnosis of IPF 
 
Histopathological Pattern 
HRCT Pattern 
Not Available 
UIP 
Probable UIP 
Possible UIP/ 
Nonclassifiable 
Fibrosis 
Not UIP 
UIP  
ELIGIBLE 
ELIGIBLE 
ELIGIBLE 
ELIGIBLE 
NOT Eligible 
Possible UIP 
NOT Eligible 
ELIGIBLE 
ELIGIBLE 
NOT Eligible 
NOT Eligible 
Inconsistent with 
UIP Pattern 
NOT Eligible 
NOT Eligible 
NOT Eligible 
NOT Eligible 
NOT Eligible 
Abbreviations: HRCT = high-resolution computed tomography; UIP = usual interstitial pneumonia.  
7.1.11. 
Emergency Sponsor Contact 
In a medical emergency, a qualified healthcare provider will attend to the subject’s immediate 
clinical management, including administration of medical intervention according to the 
Standard of Care. The healthcare provider may speak with a 
Project Physician who 
is knowledgeable about the details of this trial by contacting the
 24-Hour Call 
Center. 
United States:  
 
7.2. 
Pharmacokinetic Assessments 
7.2.1. 
Pharmacokinetic Sample Collection and Processing 
Blood samples for PK analysis will be collected by venipuncture or cannulation at the times 
indicated in the Schedule of Assessments in Appendix 5. Changes to the scheduled times of 
PK assessments may be proposed based on emerging PK data. Final determination for 
revision of the PK schedule will be the responsibility of the Sponsor and the decision will be 
documented in the TMF.  
Samples taken from subjects who received placebo will not be analyzed. 
Procedures for collection, processing, and shipping of PK blood samples will be detailed in a 
separate document.  
7.2.2. 
Pharmacokinetic Analytical Methodology 
Blood concentrations of TRK-250 will be determined using validated analytical procedures. 
Specifics of the analytical methods will be provided in separate documents. 
 
 
 
 
 
 
Page 56 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 47 of 65 
 
  
 
 
 
  
 
 
 
 
7
1. 
B
) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 57 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 48 of 65 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
   
 
 
 
 
 
 
 
 
8. SAMPLE SIZE AND DATA ANALYSIS 
8.1. 
Determination of Sample Size 
No formal statistical assessment, in terms of sample size, has been conducted. However, the 
number of subjects in each part of the present study is common in early clinical 
pharmacology studies and is considered sufficient to achieve the objectives of the study. 
8.2. 
Analysis Populations 
8.2.1. 
Safety Population 
The safety population will include all subjects who received at least 1 dose of study treatment 
(TRK-250 or placebo) and have at least 1 postdose safety assessment. 
Page 58 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 49 of 65 
8.2.2. 
Pharmacokinetic Population 
The PK population will include all subjects who received at least 1 dose of TRK-250 and 
have evaluable PK data.  
 
 
 
 
8.3. 
Safety Analysis 
Safety parameters will be listed and summarized using descriptive statistics. No inferential 
statistical analysis of safety data is planned. Each AE will be coded using the Medical 
Dictionary for Regulatory Activities (MedDRA). 
8.4. 
Pharmacokinetic Analyses 
Noncompartmental PK analysis will be performed on individual blood concentration data, 
using commercial software such as Phoenix® WinNonlin®. Blood concentrations of TRK-250 
and PK parameters will be listed and summarized using descriptive statistics.  
 
 
 
 
 
8.6. 
Interim Analysis 
Between each dose escalation, the DMC will review all available safety data to ensure it is 
safe to proceed with the planned dose escalation, and also to make recommendations on the 
escalation to a higher exposure dosing regimen, progression to Part B, and dosing regimen to 
be used in Part B. Any clinically significant results will be discussed with the Sponsor before 
dose escalation continues. Interim PK data, where available, may also be reviewed in terms 
of dose escalation. These analyses will be performed with soft locked (not hard locked) data. 
9. REFERENCES
1. King, TE. UpToDate. Clinical manifestations and diagnosis of idiopathic pulmonary
fibrosis. Available at: https://www.uptodate.com/contents/clinical-manifestations-and-
diagnosis-of-idiopathic-pulmonary-fibrosis. Accessed January 23, 2018.
2. Raghu G, Collard HR, Egan JJ, et al. ATS/ERS/JRS/ALAT Committee on Idiopathic
Pulmonary Fibrosis. An official ATS/ERS/JRS/ALAT statement: idiopathic pulmonary
fibrosis: evidence-based guidelines for diagnosis and management. Am J Respir Crit Care
Med. 2011;183(6):788-824.
Page 59 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 50 of 65 
3. Elmufdi F, Henke CA, Perlman DM, Tomic R, Kim HJ. Novel mechanisms and treatment 
of idiopathic pulmonary fibrosis. Discov Med. 2015;20(109):145-153. 
4. Spagnolo P, Del Giovane C, Luppi F, et al. Non-steroid agents for idiopathic pulmonary 
fibrosis. Cochrane Database Syst Rev. 2010;(9):CD003134. 
5. Fernandez IE, Eickelberg O. The impact of TGF-β on lung fibrosis: from targeting to 
biomarkers. Proc Am Thorac Soc. 2012;9(3):111-116. 
 
 
 
 
 
 
 
9. Miller MR, Hankinson J, Brusasco V, et al; ATS/ERS Task Force. Standardisation of 
spirometry. Eur Respir J. 2005;26(2):319-338. 
10. Pellegrino R, Viegi G, Brusasco V, et al. Interpretative strategies for lung function tests. 
Eur Respir J. 2005;26(5):948-968. 
11. Quanjer PH, Stanojevic S, Cole TJ, et al; ERS Global Lung Function Initiative. 
Multi-ethnic reference values for spirometry for the 3-95-yr age range: the global lung 
function 2012 equations. Eur Respir J. 2012;40(6):1324-1343. 
12. Graham BL, Brusasco V, Burgos F, et al. 2017 ERS/ATS standards for single-breath 
carbon monoxide uptake in the lung. Eur Respir J. 2017;49(1):1600016. 
13. Stanojevic S, Graham BL, Cooper BG, et al; Global Lung Function Initiative TLCO 
working group; Global Lung Function Initiative (GLI) TLCO. Official ERS technical 
standards: Global Lung Function Initiative reference values for the carbon monoxide 
transfer factor for Caucasians. Eur Respir J. 2017;50(3): 1700010. 
 
 
 
Page 60 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 51 of 65 
10. APPENDICES 
Page 61 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 52 of 65 
Appendix 1: Adverse Event Reporting 
Definitions 
An adverse event (AE) is any untoward medical occurrence in a subject or clinical 
investigation subject administered a pharmaceutical product, which does not necessarily have 
a causal relationship with this treatment. An AE can therefore be any unfavorable and/or 
unintended sign (including an abnormal laboratory finding), symptom, or disease temporally 
associated with the use of a study drug in humans, whether or not related to the study drug. 
Assessment of Severity 
The Investigator will be asked to provide an assessment of the severity of the AE using the 
following categories:  
• Mild: Usually transient and may require only minimal treatment or therapeutic 
intervention. The event does not generally interfere with usual activities of daily 
living. 
• Moderate: Usually alleviated with additional specific therapeutic intervention. The 
event interferes with usual activities of daily living, causing discomfort but poses no 
significant or permanent risk of harm to the subject. 
• Severe: Interrupts usual activities of daily living, or significant affects clinical status, 
or may require intensive therapeutic intervention. 
Relationship to Study Treatment 
The Investigator will make a determination of the relationship of the AE to the study drug 
using a two-category system according to the following guidelines: 
• No reasonable possibility: Evidence exists that the AE has a clear etiology other than 
the study treatment. 
• Reasonable possibility: A temporal relationship exists between the AE onset and 
administration of the study treatment and cannot be readily explained by other 
etiologies (eg, the subject’s clinical state, concomitant therapies). 
In the causality assessment of the AE, in cases when the Investigator assesses that there is no 
reasonable possibility of a relationship between the AE and study treatment, other obvious 
suspected cause(s) of the event (eg, pre-existing condition, underlying disease, intercurrent 
illness, concomitant medication, study participation, study procedures, wash-out periods) 
should be provided. 
Follow-up of Adverse Events 
Serious adverse events (SAE) will be followed up until the subject’s status reverts to 
baseline, the AE is determined by the Investigator to be irreversible, or death of the subject. 
Non-serious AE will be followed up until resolution or the AE is determined by the 
Investigator to be irreversible. This will be completed at the Investigator’s discretion. If not 
possible to follow up an SAE or AE, appropriate reasons should be provided. 
Page 62 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 53 of 65 
Adverse Drug Reactions 
All noxious and unintended responses to an investigational medicinal product (IMP) (ie, 
where a causal relationship between an IMP and an AE is at least a reasonable possibility) 
related to any dose should be considered adverse drug reactions. 
An unexpected adverse drug reaction is defined as an adverse reaction, the nature or severity 
of which is not consistent with the applicable product information (eg, Investigator’s 
Brochure for an unapproved IMP). 
Serious Adverse Events 
An SAE is defined as any untoward medical occurrence that at any dose either: 
• results in death 
• is life-threatening 
• requires inpatient hospitalization or prolongation of existing hospitalization 
• results in persistent or significant disability/incapacity (disability is defined as a 
substantial disruption of a person’s ability to conduct normal life functions) 
• results in a congenital anomaly/birth defect 
• results in an important medical event (see below). 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered SAE when, based upon appropriate medical judgement, 
they may jeopardize the subject and may require medical or surgical intervention to prevent 
one of the outcomes listed in this definition. 
The investigator should notify the Sponsor or pharmacovigilance provider assigned by the 
Sponsor of any death or SAE that he/she may become aware of occurring at any time after a 
subject has discontinued or terminated study participation and that may reasonably be related 
to this study. 
Definition of Life-Threatening 
An AE is life-threatening if the subject was at immediate risk of death from the event as it 
occurred (ie, does not include a reaction that might have caused death if it had occurred in a 
more serious form). For instance, drug-induced hepatitis that resolved without evidence of 
hepatic failure would not be considered life-threatening even though drug-induced hepatitis 
can be fatal. 
Definition of Hospitalization 
Adverse events requiring hospitalization should be considered serious. In general, 
hospitalization signifies that the subject has been detained (usually involving an overnight 
stay) at the hospital or emergency ward for observation and/or treatment that would not have 
been appropriate at the study site. When in doubt as to whether hospitalization occurred or 
was necessary, the AE should be considered as serious. 
Page 63 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 54 of 65 
Hospitalization for elective surgery or routine clinical procedures, which are not the result of 
an AE, need not be considered AE and should be recorded on a Clinical Assessment Form 
and added to the electronic Case Report Form. If anything untoward is reported during the 
procedure, this must be reported as an AE and either ‘serious’ or ‘nonserious’ attributed 
according to the usual criteria. 
Serious Adverse Event Reporting 
If an SAE occurs, the Investigators will take appropriate action immediately and will strive to 
identify the causes of the event. 
All SAE, irrespective of their causality, will be notified by the Investigator to the 
pharmacovigilance provider assigned by the Sponsor and the Medical Monitor in writing 
using the “Serious Adverse Event Report Form” (eg, by email or facsimile) within 24 hours 
of when an SAE is first recognized or reported. 
Email
 
Fax:
 
The provided information shall contain as much detail regarding the event as is available at 
the time. Investigators shall not wait to receive additional information to fully document the 
event, before notifying the SAE at the initial notification. Subsequently, a full written report 
of the SAE will be sent to the pharmacovigilance provider and Medical Monitor within 
3 working days of the original notification. Further follow-up reports will be provided when 
new information becomes available. Where applicable, information from relevant hospital 
records or autopsy reports should be obtained, if possible, and provided to the 
pharmacovigilance provider assigned by the Sponsor and the Medical Monitor. 
Food and Drug Administration (FDA)-reportable SAE will be reported to FDA by the 
pharmacovigilance provider on behalf of the Sponsor using FDA Form 3500A or equivalent 
within the timeframe required by the FDA.  
The responsible Institutional Review Board (IRB) will be notified of any FDA-reportable 
SAE within the timeframe required by the IRB. The IRB Serious and Unexpected Adverse 
Experience Submission Form will be completed and submitted with the copy of the written 
confirmation or summary of the AE. 
Pregnancy 
Pregnancy in itself is not considered an AE, unless there is a suspicion that the IMP may have 
interfered with the effectiveness of a contraceptive medication. However, complications of 
pregnancy and abnormal outcomes of pregnancy are considered AE, and many may meet 
criteria for an SAE. Complications of pregnancy and abnormal outcomes of pregnancy, such 
as ectopic pregnancy, spontaneous abortion, intrauterine fetal demise, neonatal death, or 
congenital anomaly, would meet the criteria of an SAE and therefore should be reported as an 
SAE. Elective abortions without complications should not be handled as an AE. 
All pregnancies (even if suspected) must be reported on the appropriate CRF page 
(Pregnancy Form) within 24 hours of the time that the investigator became aware of the 
Page 64 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 55 of 65 
pregnancy (or suspected pregnancy). In any case, each pregnancy should be followed until its 
termination (either by birth of a child or abortion) and the CRF Pregnancy Form should be 
updated. 
 
Page 65 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 56 of 65 
Appendix 2: Clinical Laboratory Evaluations 
Clinical chemistry: 
Hematology: 
Urinalysis: 
Alanine aminotransferase 
Albumin 
Alkaline phosphatase 
Aspartate aminotransferase 
Blood urea nitrogen  
Calcium 
Chloride 
Cholesterol 
Creatinine 
Gamma-glutamyl transferase 
Glucose 
Potassium  
Sodium  
Total bilirubina 
Total CO2 (measured as 
bicarbonate) 
Total protein 
Uric acid 
Hematocrit  
Hemoglobin 
Mean cell hemoglobin 
Mean cell hemoglobin 
concentration 
Mean cell volume 
Platelet count 
Red blood cell (RBC) count 
White blood cell (WBC) count 
WBC differential: 
   Basophils 
   Eosinophils 
   Lymphocytes 
   Monocytes 
   Neutrophils 
 
Bilirubin  
Blood 
Color and appearance  
Glucose 
Ketones 
Leukocyte esterase  
Nitrite  
pH 
Protein 
Specific gravity 
Urobilinogen 
Microscopic examination (if 
protein, leukocyte esterase, nitrite, 
or blood is positive)  
 
Serologyb: 
Drug screenc: 
Hormone panel - women only: 
Hepatitis B surface antigen 
Hepatitis C antibody 
Human immunodeficiency (HIV-1 
and HIV-2) antibodies 
 
Including but not limited to:  
Amphetamines/methamphetamines 
Barbiturates 
Benzodiazepines 
Cocaine (metabolite) 
Methadone 
Phencyclidine 
Opiates 
Tetrahydrocannabinol/ 
cannabinoids 
Follicle-stimulating hormone 
(postmenopausal women only)b  
Pregnancy test (human chorionic 
gonadotropin)c 
 
a Direct bilirubin will be analyzed if total bilirubin is elevated.  
b Only analyzed at Screening. 
c Blood pregnancy testing will be performed at Screening and Safety Follow-up visit and urine pregnancy testing will be 
performed at Day 1 predose in women of childbearing potential only. If there is doubt over the accuracy of a positive urine 
pregnancy test result, a blood pregnancy test must be obtained through the central laboratory to confirm the result. A local 
blood pregnancy test may also be performed if considered to be in the best interest of the subject (eg, travel planning). 
 
 
Page 66 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 57 of 65 
Appendix 3: Total Blood Volume 
The maximum blood volume to be withdrawn per subject (Part A or Part B), including all 
scheduled blood draws and any unscheduled blood draws that may be needed for subject 
safety, will not exceed 
mL. 
Page 67 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 58 of 65 
Appendix 4: Regulatory, Ethical, and Study Oversight Considerations 
Regulatory and Ethical Considerations 
This study will be conducted in accordance with the protocol and with the following: 
• Consensus ethical principles derived from international guidelines including the 
Declaration of Helsinki and Council for International Organizations of Medical 
Sciences (CIOMS) International Ethical Guidelines. 
• Applicable International Conference on Harmonisation (ICH) Good Clinical Practice 
(GCP) Guidelines. 
• Applicable laws and regulations. 
The protocol, protocol amendments, Informed Consent Form (ICF), Investigator Brochure, 
and other relevant documents (eg, advertisements) must be submitted to an Institutional 
Review Board (IRB) by the Investigator and reviewed and approved by the IRB before the 
study is initiated.  
Any amendments to the protocol will require IRB and regulatory authority (as locally 
required) approval before implementation of changes made to the study design, except for 
changes necessary to eliminate an immediate hazard to study subjects.  
The Investigator will be responsible for the following: 
• Providing written summaries of the status of the study to the IRB annually or more 
frequently in accordance with the requirements, policies, and procedures established 
by the IRB. 
• Notifying the IRB of serious adverse events or other significant safety findings as 
required by IRB procedures. 
• Providing oversight of the conduct of the study at the site and adherence to 
requirements of 21 Code of Federal Regulations (CFR), ICH guidelines, the IRB, 
European regulation 536/2014 for clinical studies (if applicable), and all other 
applicable local regulations. 
Finances and Insurance 
Financing and insurance will be addressed in a separate agreement. 
Informed Consent 
Prior to starting participation in the study, each subject will be provided with a study-specific 
ICF giving details of the study drugs, procedures, and potential risks of the study. Subjects 
will be instructed that they are free to obtain further information from the Investigator and 
that their participation is voluntary and they are free to withdraw from the study at any time. 
Subjects will be given an opportunity to ask questions about the study prior to providing 
consent for participation. 
Page 68 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 59 of 65 
Following discussion of the study with site personnel, subjects will sign 2 copies of the ICF 
in the presence of a suitably trained member of staff to indicate that they are freely giving 
their informed consent. One copy will be given to the subject, and the other will be 
maintained in the subject’s records. 
Subjects must be re-consented to the most current version of the ICF(s) during their 
participation in the study. 
Subject Data Protection 
Subjects will be assigned a unique identifier and will not be identified by name in Case 
Report Forms (CRF), study-related forms, study reports, or any related publications. Subject 
and Investigator personal data will be treated in compliance with all applicable laws and 
regulations. In the event the study protocol, study report, or study data are included in a 
public registry, all identifiable information from individual subjects or Investigators will be 
redacted according to applicable laws and regulations.  
The subject must be informed that his/her personal study-related data will be used by the 
Sponsor in accordance with local data protection law. The level of disclosure must also be 
explained to the subject. The subject must also be informed that his/her study-related data 
may be examined by Sponsor or Contract Research Organization (CRO) auditors or other 
authorized personnel appointed by the Sponsor, by appropriate IRB members, and by 
inspectors from regulatory authorities. 
Disclosure 
All information provided regarding the study, as well as all information collected and/or 
documented during the course of the study, will be regarded as confidential. The Investigator 
agrees not to disclose such information in any way without prior written permission from the 
Sponsor. 
Data Quality Assurance 
The following data quality steps will be implemented: 
• All subject data relating to the study will be recorded on CRF unless directly 
transmitted to the Sponsor or designee electronically (eg, laboratory data). The 
Investigator is responsible for verifying that data entries are accurate and correct by 
electronically signing the CRF.  
• The Investigator must maintain accurate documentation (source data) that supports the 
information entered in the CRF.  
• The Investigator must permit study-related monitoring, audits, IRB review, and 
regulatory agency inspections and provide direct access to source data documents.  
• The Sponsor or designee is responsible for the data management of this study 
including quality checking of the data. Predefined, agreed risks, monitoring 
thresholds, quality tolerance thresholds, controls, and mitigation plans will be 
documented in a risk management register. Additional details of quality checking to 
be performed on the data may be included in a Data Management Plan.  
Page 69 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 60 of 65 
• A Study Monitor will perform ongoing source data verification to confirm that data 
entered into the CRF by authorized site personnel are accurate, complete, and 
verifiable from source documents; that the safety and rights of subjects are being 
protected; and that the study is being conducted in accordance with the currently 
approved protocol and any other study agreements, ICH GCP, and all applicable 
regulatory requirements. 
• Records and documents, including signed ICFs, pertaining to the conduct of this study 
must be retained by the Investigator in accordance with 21 CFR 312.62(c) unless local 
regulations or institutional policies require a longer retention period. No records may 
be destroyed during the retention period without the written approval of the Sponsor. 
No records may be transferred to another location or party without written notification 
to the Sponsor.  
Investigator Documentation Responsibilities 
All individual, subject-specific study data will also be entered into a 21 CFR Part 
11-compliant electronic data capture (EDC) system on a CRF in a timely fashion. 
All data generated from external sources (eg, laboratory and bioanalytical data), and 
transmitted to the Sponsor or designee electronically, will be integrated with the subject’s 
CRF data in accordance with the Data Management Plan.  
A CRF must be completed for each subject who signs an ICF and undergoes any screening 
procedures, according to the CRF completion instructions. The Sponsor, or CRO, will review 
the supporting source documentation against the data entered into the CRF to verify the 
accuracy of the electronic data. The Investigator will ensure that corrections are made to the 
CRF and that data queries are resolved in a timely fashion by the study staff. 
The Investigator will sign and date the CRF via the EDC system’s electronic signature 
procedure. These signatures will indicate that the Investigator reviewed and approved the 
data on the CRF, data queries, and site notifications. 
Publications 
If on completion of the study the data warrant publication, the Investigator may publish the 
results in recognized (refereed) scientific journals subject to the provisions of the clinical 
study agreement (CSA). Unless otherwise specified in the CSA, the following process shall 
occur: 
If the Investigator expects to participate in the publication of data generated from this site, he 
or she must obtain prior approval of the Sponsor before submission for publication or 
presentation. 
Page 70 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 61 of 65 
Appendix 5: Schedule of Assessments 
Page 71 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 62 of 65 
Table 1: 
Schedule of Assessments – Part A (Single Dose) 
Study Epoch 
Screening 
Treatment 
Follow-up 
Study Procedures 
Screening 
Day 1 predose 
Day 1 dosing & 
postdose 
Days 2 & 3 
Safety 
Follow-up  
Additional 
Follow-up  
Study Day (Window) 
-28 to -1 
1 
1 
2 & 3 
7 (+3)  
14 (±3) 
Informed consent 
X 
 
 
 
 
 
Inclusion/exclusion criteria 
X 
X 
 
 
 
 
Demographic dataa 
X 
 
 
 
 
 
Medical history 
X 
Xb 
 
 
 
 
Body weight and height 
X 
 
 
 
Xc 
 
Oral body temperature 
X 
Xd 
X 
X 
X 
X 
Blood pressure and pulse rate 
X 
Xd 
Xe 
X 
X 
X 
SpO2 
X 
Xd 
Xe 
X 
X 
X 
Physical examination 
X 
X 
Xf 
Xf 
Xf 
Xf 
Pregnancy testg 
X 
X 
 
 
X 
 
Follicle-stimulating hormoneh 
X 
 
 
 
 
 
12-lead ECG 
X 
X 
Xi 
X 
X 
 
HRCT 
Xj 
 
 
 
 
 
Spirometryk 
X 
Xl 
Xl 
X 
X 
X 
Carbon monoxide diffusion capacityk 
X 
Xd 
 
X 
X 
X 
Clinical chemistry and hematology 
X 
X 
 
Day 3 only 
X 
 
Serology 
X 
 
 
 
 
 
Urine drugs of abuse screenm 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
Nonresidential visit 
X 
 
 
Day 3 only 
X 
X 
Study treatment administration 
 
 
X 
 
 
 
Adverse event recording 
 
X 
X 
X 
X 
X 
 
 
 
 
 
 
 
Pharmacokinetic blood sampling 
 
X 
Xo 
 
 
 
 
 
 
 
 
 
 
Note, where multiple assessments are required at a single timepoint, the following order of procedures should be used: blood sampling, spirometry, other procedures (ECGs should be 
scheduled before vital signs measurements). See also footnote k. 
Note, timing of safety and pharmacokinetic assessments may be changed based upon findings from previous cohorts. 
Abbreviations: ECG = electrocardiogram; HRCT = high-resolution computed tomography; SpO2 = oxygen saturation. 
Page 72 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 63 of 65 
a To include smoking history. 
b Interim medical history. 
c Height measured only at Screening. 
d Within 60 minutes prior to dosing. 
e 1 hour and 2 hours after completion of dose administration. 
f Symptom-directed physical examination. 
g In women of childbearing potential. Blood pregnancy test will be performed at Screening and Safety Follow-up; urine pregnancy test will be performed at Day 1. If the Day 1 pregnancy 
test is positive, dosing of that subject should not proceed. If there is doubt over the accuracy of a positive urine pregnancy test result, a blood pregnancy test must be obtained through the 
central laboratory to confirm the result. A local blood pregnancy test may also be performed if considered to be in the best interest of the subject (eg, travel planning). If the local test is 
negative and there are no other clinical findings suggestive of pregnancy, the subject may be dosed. 
h In postmenopausal women (at least 12 months postcessation of menses without an alternative medical cause) only, to confirm postmenopausal status. Confirmation of postmenopausal 
status requires level >40 mIU/mL in addition to appropriate history (refer to Section 6.5). 
i 2 hours after completion of dose administration. 
j If the most recent historical HRCT was performed more than 12 months prior to the initiation of Screening OR if the HRCT performed within 12 months prior to the initiation of 
Screening is determined not to be of suitable quality by the central over-reader, the imaging study must be repeated during Screening. Any HRCT used to establish subject eligibility must 
undergo central interpretation. 
k Spirometry should preferably be performed before carbon monoxide diffusion capacity assessments at timepoints where both assessments are required, but if not feasible, the rationale 
should be documented in source records and the order of testing implemented on Day 1 should be used at every subsequent study visit. 
l Completion of a minimum of 3, maximum of 6 forced expiratory maneuvers within 60 minutes predose. Preferably 3 but a minimum of 2 maneuvers as soon as feasible but within 
15 minutes after completion of dose administration and at 30 ± 5 minutes, 60 ± 10 minutes, and 90 ± 15 minutes after completion of dose administration (up to a maximum of 16 forced 
expiratory maneuvers per day). 
m Exclusionary only if positive for a drug or metabolite of a drug that has not been prescribed and that remains positive on repeat testing. 
 
 
 
o Within 5 minutes after completion of dose administration and 10, 30, and 60 minutes after completion of dose administration, to assay for parent drug (TRK-250). PK sampling times are 
given as targets to be achieved within reasonable limits (eg, ±10%). 
 
 
 
Page 73 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 64 of 65 
Table 2:  
Schedule of Assessments – Part B (Multiple Dose) 
Study Epoch 
Screening 
Treatment 
Follow-up 
Study Procedures 
Screening 
Days 1, 8, 15 & 22 
predose 
Days 1, 8, 15 & 22 
dosing & postdose 
Safety Follow-up  
Additional 
Follow-up 
Study Day (Window) 
-28 to -1 
1, 8 (+1), 15 (+1) & 22 (+1) 
(≥7 days between doses) 
29 (+3) 
36 (±3) 
Informed consent 
X 
 
 
 
 
Inclusion/exclusion criteria 
X 
X 
 
 
 
Demographic dataa 
X 
 
 
 
 
Medical history 
X 
Xb 
 
 
 
Body weight and height 
X 
 
 
Xc 
 
Oral body temperature 
X 
Xd 
X 
X 
X 
Blood pressure and pulse rate 
X 
Xd 
Xe 
X 
X 
SpO2 
X 
Xd 
Xe 
X 
X 
Physical examination 
X 
X 
Xf 
Xf 
Xf 
Pregnancy testg 
X 
Day 1 predose only 
 
X 
 
Follicle-stimulating hormoneh 
X 
 
 
 
 
12-lead ECG 
X 
X 
Xi 
X 
 
HRCT 
Xj 
 
 
 
 
Spirometryk 
X 
Xl 
Xl 
X 
X 
Carbon monoxide diffusion capacityk 
X 
Xd 
 
X 
X 
Clinical chemistry and hematology 
X 
X 
 
X 
 
Serology 
X 
 
 
 
 
Urine drugs of abuse screenm 
X 
 
 
 
 
 
 
 
 
 
 
Study treatment administration 
 
 
X 
 
 
Adverse event recording 
 
X 
X 
X 
X 
 
 
 
 
 
 
Pharmacokinetic blood sampling 
 
Days 1 & 22 
Days 1 & 22n 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Page 74 of 75


Protocol Amendment 3 
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
Final Protocol Amendment 3, 24 September 2018 
Page 65 of 65 
Study Epoch 
Screening 
Treatment 
Follow-up 
Study Procedures 
Screening 
Days 1, 8, 15 & 22 
predose 
Days 1, 8, 15 & 22 
dosing & postdose 
Safety Follow-up  
Additional 
Follow-up 
Study Day (Window) 
-28 to -1 
1, 8 (+1), 15 (+1) & 22 (+1) 
(≥7 days between doses) 
29 (+3) 
36 (±3) 
 
 
 
 
 
 
 
 
Note, where multiple assessments are required at a single timepoint, the following order of procedures should be used: blood sampling, spirometry, other procedures (ECGs should be 
scheduled before vital signs measurements). 
. See also footnote k. 
Note, timing of safety and pharmacokinetic assessments may be changed based upon findings from previous cohorts. 
Abbreviations: 
 DLCO = carbon monoxide diffusion capacity; ECG = electrocardiogram; HRCT = high-resolution computed tomography; 
 SpO2 = oxygen saturation. 
a To include smoking history. 
b Interim medical history. 
c Height measured only at Screening. 
d Within 60 minutes prior to dosing. 
e 1 hour and 2 hours after completion of dose administration. 
f Symptom-directed physical examination. 
g In women of childbearing potential. Blood pregnancy test will be performed at Screening and Safety Follow-up; urine pregnancy test will be performed at Day 1. If the Day 1 pregnancy 
test is positive, dosing of that subject should not proceed. If there is doubt over the accuracy of a positive urine pregnancy test result, a blood pregnancy test must be obtained through the 
central laboratory to confirm the result. A local blood pregnancy test may also be performed if considered to be in the best interest of the subject (eg, travel planning). If the local test is 
negative and there are no other clinical findings suggestive of pregnancy, the subject may be dosed. 
h In postmenopausal women (at least 12 months postcessation of menses without an alternative medical cause) only, to confirm postmenopausal status. Confirmation of postmenopausal 
status requires level >40 mIU/mL in addition to appropriate history (refer to Section 6.5). 
i 2 hours after completion of dose administration. 
j If the most recent historical HRCT was performed more than 12 months prior to the initiation of Screening OR if the HRCT performed within 12 months prior to the initiation of 
Screening is determined not to be of suitable quality by the central over-reader, the imaging study must be repeated during Screening. Any HRCT used to establish subject eligibility must 
undergo central interpretation. 
k Spirometry should preferably be performed before DLCO at timepoints where both assessments are required, but if not feasible, the rationale should be documented in source records and 
the order of testing implemented on Day 1 should be used at every subsequent study visit. 
l Completion of a minimum of 3, maximum of 6 forced expiratory maneuvers within 60 minutes predose. Preferably 3 but a minimum of 2 maneuvers as soon as feasible but within 
15 minutes after completion of dose administration and at 30 ± 5 minutes, 60 ± 10 minutes, and 90 ± 15 minutes after completion of dose administration (up to a maximum of 16 forced 
expiratory maneuvers per day). 
m Exclusionary only if positive for a drug or metabolite of a drug that has not been prescribed and that remains positive on repeat testing. 
n Within 5 minutes after completion of dose administration and 10, 30, and 60 minutes after completion of dose administration, to assay for parent drug (TRK-250). PK sampling times are 
given as targets to be achieved within reasonable limits (eg, ±10%). 
 
 
p To avoid artifacts in any subsequent spirometry or DLCO assessments, 
 
  
r The Day 1 sample must be taken prior to administration of any study treatment. Although it is preferred that the Day 15 sample is taken predose, this is not mandatory. 
 
Page 75 of 75

SAP_001.pdf: 
Final: 25 June 2019 
Page 1 of 17 
STATISTICAL ANALYSIS PLAN 
TRK-250 – A Phase I, Double-Blind, Placebo-Controlled, Single and 
Multiple Inhaled Dose, Safety, Tolerability, and Pharmacokinetic Study 
of TRK-250 in Subjects with Idiopathic Pulmonary Fibrosis 
Statistical Analysis Plan Status: Final 
Statistical Analysis Plan Date: 25 June 2019 
Study Drug: TRK-250 
Sponsor Reference Number: 250IPF01 
 Study Number: 8376269 
Clinical Phase I 
Sponsor: 
Toray Industries, Inc. 
1-1, Nihonbashi-muromachi 2-
chome, 
Chuo-ku, Tokyo, 103-8666 Japan 
Study Site: 
Multiple Sites 
: 
 
 
 
: 
 
 
 
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 2 of 17 
1  STATISTICAL ANALYSIS PLAN APPROVAL SIGNATURES 
By signing this page when the Statistical Analysis Plan (SAP) is considered final, the signatories 
agree to the statistical, pharmacokinetic (PK)
 analyses to be 
performed for this study, and to the basic format of the tables, figures, and listings (TFLs). Once 
the SAP has been signed, programming of the TFLs based upon this document can proceed. Any 
modifications to the SAP and TFLs made after signing may result in a work-scope change. 
 
 
 
 
 
 
 
 
 
 
 
 
Date 
 
 
 
 
 
 
 
 
 
 
Date 
 
 
Sponsor approval: 
 
 
 
 
 
 
 
 
 
Date 
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 3 of 17 
2  TABLE OF CONTENTS 
1 
STATISTICAL ANALYSIS PLAN APPROVAL SIGNATURES ................................... 2 
2 
TABLE OF CONTENTS ................................................................................................... 3 
3 
ABBREVIATIONS ............................................................................................................ 5 
4 
INTRODUCTION .............................................................................................................. 6 
5 
STUDY OBJECTIVES AND ENDPOINTS ...................................................................... 6 
5.1 
Primary ........................................................................................................................ 6 
5.2 
Secondary .................................................................................................................... 6 
 
 
5.4 
Endpoints ..................................................................................................................... 7 
6 
STUDY DESIGN ............................................................................................................... 8 
6.1 
Part A ........................................................................................................................... 8 
6.2 
Part B ........................................................................................................................... 8 
7 
TREATMENTS .................................................................................................................. 9 
8 
SAMPLE SIZE JUSTIFICATION ................................................................................... 10 
9 
DEFINITION OF ANALYSIS POPULATIONS ............................................................ 10 
10 STATISTICAL METHODOLOGY ................................................................................. 11 
10.1 General ....................................................................................................................... 11 
10.1.1 Definition of Baseline and Change from Baseline ........................................... 11 
10.1.2 Repeat and Unscheduled Readings .................................................................. 11 
10.2 Demographics and Subject Disposition ..................................................................... 12 
10.3 Pharmacokinetic Assessment ..................................................................................... 12 
10.3.1 Pharmacokinetic Analysis ................................................................................ 12 
10.3.2 Criteria for Handling Concentrations Below the Limit of Quantification in 
Pharmacokinetic Analysis ................................................................................ 13 
10.3.3 Calculation of AUC .......................................................................................... 13 
10.3.4 Anomalous Values ........................................................................................... 13 
10.4 Presentation of Pharmacokinetic Data ....................................................................... 14 
10.4.1 
Presentation of Pharmacokinetic Concentration Data ...................................... 14 
10.4.2 
Presentation of Pharmacokinetic Parameters ................................................... 14 
10.4 Pharmacokinetic Statistical Methodology ................................................................. 14 
 
 
10.6 Safety and Tolerability Assessments ......................................................................... 15 
10.6.1 Adverse Events ................................................................................................. 15 
10.6.2 Clinical Laboratory Parameters ........................................................................ 15 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 4 of 17 
10.6.3 Vital Signs ........................................................................................................ 15 
10.6.4 Electrocardiogram ............................................................................................ 15 
10.6.5 Pulse Oximetry ................................................................................................. 16 
10.6.6 Spirometry ........................................................................................................ 16 
10.6.7 Carbon Monoxide Diffusion Capacity Test (DLCO) ....................................... 16 
10.6.8 Other Assessments ........................................................................................... 16 
10.6.9 Safety and Tolerability Statistical Methodology .............................................. 16 
11 INTERIM ANALYSES .................................................................................................... 16 
12 CHANGES FROM THE PROTOCOL SPECIFIED STATISTICAL ANALYSES ....... 16 
13 DATA PRESENTATION ................................................................................................ 17 
13.1 Insufficient Data for Presentation ............................................................................... 17 
14 REFERENCES ................................................................................................................. 17 
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 5 of 17 
 
3  ABBREVIATIONS 
Abbreviations pertain to the SAP only (not the TFLs). 
AE 
adverse event 
AUC0-tlast 
area under the concentration-time curve from time zero to the time 
of the last quantifiable concentration 
Cmax 
maximum observed plasma concentration 
BID 
twice daily 
DLCO 
carbon monoxide diffusion capacity 
DMC 
data monitoring committee  
ECG 
electrocardiogram 
FEV1 
Forced expiratory volume over 1 second 
FVC 
forced vital capacity 
ICF 
informed consent form 
IPF 
idiopathic pulmonary fibrosis 
MedDRA 
Medical Dictionary for Regulatory Activities 
 
 
PK 
pharmacokinetic 
QTcB 
QT interval corrected for heart rate using Bazett’s method 
QTcF 
QT interval corrected for heart rate using Fridericia's method 
SAP 
Statistical Analysis Plan 
SOC 
system organ class 
SPO2 
Oxygen saturation 
TEAE 
treatment-emergent adverse event 
TFLs 
tables, figures, and listings 
Tmax 
time of maximum observed concentration 
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 6 of 17 
4  INTRODUCTION 
This SAP has been developed after review of the clinical study protocol (Final Version dated 01 
June 2019), protocol amendment No. 1 (dated 17 July 2019), protocol amendment No. 2 (dated 
24 July 2018) and protocol amendment No. 3 (dated 24 September 2018). 
This SAP describes the planned analysis of the safety, tolerability, PK
 from this 
study. A detailed description of the planned TFLs to be presented in the Clinical Study Report 
(CSR) is provided in the accompanying TFL shell document. 
The intent of this document is to provide guidance for the statistical analyses of PK
. 
In general, the analyses are based on information from the protocol, unless they have been 
modified by agreement between Toray Industries, Inc. and 
 
. A limited amount of information concerning this study (eg, objectives, study design) 
is given to help the reader’s interpretation. This SAP must be finalised prior to the lock of the 
clinical database for this study. When the SAP and TFL shells are agreed upon and finalized, 
they will serve as the template for this study’s CSR. 
This SAP supersedes the statistical considerations identified in the protocol; where 
considerations are substantially different, they will be so identified. If additional analyses are 
required to supplement the planned analyses described in this SAP, they may be performed and 
will be identified in the CSR. Any substantial deviations from this SAP will be agreed upon 
between Toray Industries, Inc. and 
 and identified in the CSR. 
This SAP is written with consideration of the recommendations outlined in the International 
Conference on Harmonisation (ICH) E9 guideline entitled, “Guidance for Industry: Statistical 
Principles for Clinical Trials” and the ICH E3 guideline entitled, “Guidance for Industry: 
Structure and Content of Clinical Study Reports.”1,2 
5  STUDY OBJECTIVES AND ENDPOINTS 
5.1 
Primary 
The primary objective of the study is to assess the safety and tolerability of single and multiple 
inhaled doses of TRK-250 in subjects with idiopathic pulmonary fibrosis (IPF). 
5.2 
Secondary 
The secondary objective of the study is to assess the PK in blood following single and multiple 
inhaled doses of TRK-250 in subjects with IPF. 
 
 
 


Statistical Analysis Plan  
CONFIDENTIAL 
 
Sponsor Reference: 250IPF01 
 
 
 
 
5.4 
Endpoints 
The primary safety endpoints for this study are as follows: 
• body weight
• vital signs measurements
• oxygen saturation (SpO2) by pulse oximetry
• hematology, clinical chemistry, and urinalysis test results
• 12-lead electrocardiogram (ECG) parameters
• forced expiratory volume over 1 second (FEV1), forced vital capacity (FVC)
• incidence and severity of adverse events (AE)
The single and multiple ascending dose PK outcome endpoints of TRK-250 are as follows 
(where applicable):  
• area under the concentration-time curve (AUC) from time zero to the time of the last
quantifiable concentration (AUC0-tlast) 
• maximum observed concentration (Cmax)
• time to Cmax (Tmax)
Other PK parameters may also be added.  
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 8 of 17 
 
  
6  STUDY DESIGN 
This will be a Phase I, double-blind, randomized, placebo-controlled study conducted in 2 parts, 
single and multiple inhaled dose, at multiple sites. 
6.1 
Part A 
Part A will comprise a single-dose, single-period, sequential-cohort study. Potential subjects will 
be screened to assess their eligibility to enter the study within 28 days prior to study drug 
administration. 
It is planned that 16 subjects will be studied in 4 cohorts (Cohorts A1 to A4), each consisting of 
4 subjects. Following DMC review of the safety and tolerability data from the first 4 cohorts, up 
to 3 further optional cohorts (Cohorts A5 to A7) of at least 4 subjects may be added to Part A to 
evaluate additional doses or to further evaluate planned doses. 
 
6.2 
Part B 
Part B will comprise a multiple-dose, single-period, sequential-cohort study. Potential subjects 
will be screened to assess their eligibility to enter the study within 28 days prior to initial study 
drug administration. 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 9 of 17 
It is planned to study 18 subjects in 3 cohorts (Cohorts B1 to B3), each consisting of 6 subjects. 
Up to 3 further optional cohorts (Cohorts B4 to B6) of at least 6 subjects may be added to 
evaluate additional doses or to further evaluate planned doses. 
 
 
7  TREATMENTS 
The following is a list of the masked study treatment abbreviations and ordering that will be used 
in the Closed Report. 
Part A: 
Cohorts 
Treatment Order on TFLs 
Cohort A1 
TRK-250 2 mg 
Cohort A2 
TRK-250 10 mg 
Cohort A3 
TRK-250 30 mg 
Cohort A4 
TRK-250 60 mg 
Cohort A... 
… mg 
All Cohorts in part A 
Part A Pooled placebo 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 10 of 17 
Up to 3 further optional cohorts (Cohorts A5 to A7) of at least 4 subjects may be added to Part A 
to evaluate additional doses or to further evaluate planned doses. 
Part B: 
Cohorts 
Treatment  Order on TFLs 
Cohort B1 
TRK-250 10 mg weekly 
Cohort B2 
TRK-250 30 mg weekly 
Cohort B3 
TRK-250 60 mg weekly 
Cohort B… 
… mg 
All Cohorts in part B 
Part B Pooled placebo 
 
Up to 3 further optional cohorts (Cohorts B4 to B6) of at least 6 subjects may be added to Part B 
to evaluate additional doses or to further evaluate planned doses. 
8  SAMPLE SIZE JUSTIFICATION 
No formal statistical assessment, in terms of sample size, has been conducted. However, the 
number of subjects in each part of the present study is common in early clinical pharmacology 
studies and is considered sufficient to achieve the objectives of the study. 
9  DEFINITION OF ANALYSIS POPULATIONS 
The Safety Population will include all subjects who received at least 1 dose of study treatment 
(TRK-250 or placebo) and have at least 1 postdose safety assessment. 
The Pharmacokinetic (PK) Population will include all subjects who received at least 1 dose of 
TRK-250 and have evaluable PK data. 
 
 
The All Subjects Population will consist of any subjects who signed informed consent and had 
study assessments recorded on the database as per the protocol. 
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 11 of 17 
10  
STATISTICAL METHODOLOGY 
10.1 
General 
Data listings will be provided for the All Subjects Population. Summary statistics and statistical 
analyses will be performed for subjects included in the relevant analysis populations 
 
For continuous data, summary statistics will include the arithmetic mean, arithmetic standard 
deviation, median, minimum, maximum, and number. For log-normal data (eg, the PK 
parameters: areas under the concentration-time curve [AUCs] and maximum observed 
concentration [Cmax]), the geometric mean and geometric coefficient of variation (CV%) will 
also be presented. For categorical data, frequency counts and percentages will be presented. Data 
listings will be provided for all subjects up to the point of withdrawal, with any subjects 
excluded from the relevant population highlighted. For the calculation of summary statistics and 
statistical analysis, unrounded data will be used. 
Missing values will not be imputed. 
Data analysis will be performed using SAS® Version 9.4 or greater. 
10.1.1 Definition of Baseline and Change from Baseline 
Baseline for each parameter is defined as the last value measured prior to dosing, including 
repeat (vital signs and electrocardiograms [ECGs]) and unscheduled (clinical laboratory 
parameters) readings (see Section 10.1.2 for definitions of repeat and unscheduled readings). For 
vital signs taken in triplicate, baseline will be the median of the last 3 values taken prior to 
dosing. For ECGs taken in triplicate, baseline will be the mean of the last 3 values taken prior to 
dosing. 
Mean change from baseline is the mean of all individual subjects’ change from baseline values. 
Each individual change from baseline will be calculated by subtracting the individual subject’s 
baseline value from the value at the timepoint. The individual subject’s change from baseline 
values will be used to calculate the mean change from baseline using a SAS procedure such as 
Proc Univariate. 
10.1.2 Repeat and Unscheduled Readings 
Repeat readings occur when the original vital signs or ECG result requires confirmation. Repeat 
readings are labelled as ‘Repeat’ in the listings and replace the original readings in all summaries 
and changes from baseline presentations and calculations. Prior to dosing, all readings taken in 
addition to the original reading are defined as predose repeats. Postdose repeat readings are 
defined as readings collected within 15 minutes of the actual time of the original reading. Where 
results are taken in triplicate and repeated, the last 3 readings are used in all subsequent 
calculations. 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 12 of 17 
With the exception of predose results described above, unscheduled readings for vital signs or 
ECGs are defined as readings collected >15 minutes from the actual time of the original reading. 
Where results are taken in triplicate, the original reading is defined as the first reading of the 
triplicate. All results not taken at a scheduled timepoint for other data types (eg, clinical 
laboratory parameters) are unscheduled. Unscheduled readings are labelled as ‘Unscheduled’ in 
the listings. Because unscheduled readings are not associated with any scheduled timepoint, they 
are excluded from all summaries (with the exception that they may be used as baseline, as stated 
in Section 10.1.1). 
10.2 
Demographics and Subject Disposition 
The demographic variables age, sex, race, ethnicity, body weight, height, and body mass index 
will be summarized and listed. Subject disposition will be summarized and listed. 
10.3 
Pharmacokinetic Assessment 
10.3.1 Pharmacokinetic Analysis 
The following pharmacokinetic parameters will be determined where possible from the whole 
blood concentrations of TRK-250 using non-compartmental methods performed using Phoenix 
WinNonlin (Version 8.1 or higher): 
Parameter 
Definition 
AUC(0-tlast) 
area under the concentration-time curve (AUC) from time zero to the last 
quantifiable concentration,  
Cmax 
maximum observed concentration 
Tmax 
time of maximum observed concentration 
 
Additional pharmacokinetic parameters may be determined where appropriate. 
Pharmacokinetic analysis will, where possible, be carried out using actual postdose times 
recorded in the raw data. If actual times are missing, nominal times may be used with sponsor 
approval.  
Concentrations are used as supplied by the analytical laboratory for PK analysis. The units of 
concentration and resulting PK parameters, with amount or concentration in the unit, will be 
presented as they are received from the analytical laboratory.  
Cmax and Tmax will be obtained directly from the plasma concentration-time profiles.   
For multiple peaks, the highest postdose concentration will be reported as Cmax. In the case that 
multiple peaks are of equal magnitude, the earliest Tmax will be reported.  


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 13 of 17 
10.3.2 Criteria for Handling Concentrations Below the Limit of Quantification in 
Pharmacokinetic Analysis 
• Concentration values that are below the limit of quantification (BLQ) will be set to zero, 
with defined exceptions as follows: 
o Any embedded BLQ value (between 2 quantifiable concentrations) and BLQ 
values following the last quantifiable concentration in a profile will be set to 
missing for the purposes of PK analysis. 
o If there are late positive concentration values following 2 BLQ concentration 
values in the apparent terminal phase, these values will be evaluated. If these 
values are considered to be anomalous, they will be set to missing. 
o If an entire concentration-time profile is BLQ, the profile will be excluded from 
the PK analysis. 
o If a predose concentration is missing, these values will be set to zero. 
10.3.3 Calculation of AUC 
• AUC values will be calculated using the linear trapezoidal rule for increasing 
concentrations and the logarithmic trapezoidal rule for decreasing concentrations. 
• 
The minimum requirement for the calculation of AUC will be the inclusion of at least 
3 consecutive whole blood concentrations above the lower limit of quantification (LLOQ), 
with at least 1 of these concentrations following Cmax. 
• 
For any partial AUC determination (if required), nominal time will generally be used for 
the end of the interval. Actual times for partial AUC intervals may be used at the 
discretion of the Pharmacokineticist. 
10.3.4 Anomalous Values 
• 
If a value is considered to be anomalous due to being inconsistent with the expected PK 
profile, it may be appropriate to exclude this point from the PK analysis. However, the 
exclusion of data must have strong justification and will be documented in the raw data 
and study report. 
• 
Embedded BLQ values may be considered anomalous depending on the route of 
administration and the characteristics of the drug. 
• 
PK parameter data associated with quantifiable predose value(s) greater than 5% of Cmax 
may be excluded from the summary statistics of PK tables and statistical analysis at the 
discretion of the Pharmacokineticist. 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 14 of 17 
10.4 Presentation of Pharmacokinetic Data 
10.4.1 Presentation of Pharmacokinetic Concentration Data 
The following rules will be applied if there are values that are BLQ or if there are missing values 
(e.g., no result [NR]) in a plasma or whole blood concentration data series to be summarized. 
o For the calculation of summary statistics, BLQ values will be set to zero. 
o If 
an 
embedded 
BLQ 
value 
is 
considered 
anomalous 
within 
the 
concentration-time profile, this value will be excluded from the summary 
statistics. 
o Where there is NR, these will be set to missing. 
o If there are less than three values in the data series, only the min, max and N will 
be presented. The other summary statistics will be denoted as not calculated (NC). 
BLQ is considered a value. 
o If all the values are BLQ, then the arithmetic mean, arithmetic SD, median, min 
and max will be presented as zero, and the geometric mean and geometric CV% 
will be denoted as NC. 
o If the value of the arithmetic mean or median is below the lower limit of 
quantification, it will be presented as zero and the geometric mean and geometric 
CV% will be denoted as NC. 
10.4.2 Presentation of Pharmacokinetic Parameters  
• 
For the calculation of summary statistics of PK parameters, all NR and NC values in a 
data series will be set to missing. 
• 
The AUC values will be set to NC if they have been calculated using fewer than three 
concentrations, and/or three concentrations if the last is Cmax. 
 
10.4 Pharmacokinetic Statistical Methodology 
Where data available blood concentration of TRK-250 and corresponding PK parameters will be 
summarized and listed. PK concentration profiles over time will be presented graphically.  
 
 
 
 
 
 
 
 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 15 of 17 
 
10.6 
Safety and Tolerability Assessments 
10.6.1 Adverse Events 
A pre-treatment adverse event (AE) is defined as an AE that starts after the subject has provided 
written informed consent and that resolves prior to the first dosing occasion, or an AE that starts 
prior to the first dosing occasion and does not increase in severity after dosing. A 
treatment-emergent AE (TEAE) is defined as an AE that occurs postdose or that is present 
predose and becomes more severe postdose. 
All AEs will be listed. The TEAEs will be summarized by study part, treatment, severity, and 
relationship to the study drug. The frequency (the number of TEAEs, the number of subjects 
experiencing a TEAE, and the percentage of subjects experiencing a TEAE) of TEAEs will be 
summarized by treatment, and by Medical Dictionary for Regulatory Activities system organ 
class and preferred term. The summary and frequency TEAE tables will be presented for all 
causalities and for those TEAEs considered having reasonable possibility relationship to the 
study drug. 
For any AEs that change severity ratings the AE will be included only once under the maximum 
severity rating in the summaries. 
All study agent-related AEs, serious adverse events (SAE), AEs leading to withdrawal, AEs of 
severe intensity, life threatening and deaths will be listed separately. 
10.6.2 Clinical Laboratory Parameters 
All clinical chemistry, hematology data will be summarized by treatment. Changes from baseline 
will be calculated. Individual clinical chemistry, hematology and urinalysis listings will also be 
presented. In addition, all clinical chemistry, hematology and urinalysis data outside the clinical 
reference ranges will be summarized and listed by parameter and treatment. 
10.6.3 Vital Signs 
The vital signs data will be summarised by treatment, together with changes from baseline.  
Corresponding individual listings will also be presented. 
10.6.4 Electrocardiogram 
The ECG data will be obtained directly from the 12-lead ECG traces. These data include the QT 
interval calculated using the Bazett correction (QTcB), the QT interval calculated using the 
Fridericia correction (QTcF), the PR and QT intervals, the QRS duration, and heart rate. ECG 
data will be summarized and listed. 
The ECG data will be summarised by treatment, together with changes from baseline. 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 16 of 17 
An outlier analysis will be performed including all individual postdose measurements (not the 
mean data), including all repeat and unscheduled readings. The frequency of subjects with a 
maximum increase from baseline in QTcB and QTcF intervals will be summarised for each 
treatment according to the following categories: >30, >60, and ≤30 ms. All incidences of >30 
and >60 ms will be flagged on the listing. In addition, the frequency of subjects with QTcB and 
QTcF postdose values will be summarised for each treatment, according to the following 
categories: >450, >480, >500, and ≤450 ms. All incidences of >450, >480, and >500 ms will be 
flagged on the listing.  
10.6.5 Pulse Oximetry 
SPO2 data and change from baseline values will also be summarized per treatment and listed for 
each subject. The baseline is the last measurement before dosing. 
10.6.6 Spirometry 
Forced expiratory volume over 1 second (FEV1), forced vital capacity (FVC) and their ratio will 
be summarized by treatment at each timepoint and listed for each individual along with their 
change from baseline values. 
10.6.7 Carbon Monoxide Diffusion Capacity Test (DLCO) 
DLCO data will be summarized by treatment at each timepoint and listed for each individual 
along with their change from baseline values. 
10.6.8 Other Assessments 
All other safety assessments,
 not detailed in this section will be 
listed but not summarized or statistically analysed. 
Medical history data will be presented. 
10.6.9 Safety and Tolerability Statistical Methodology 
No inferential statistical analyses are planned. 
11  
INTERIM ANALYSES 
No interim statistical analyses are planned. 
12  
CHANGES FROM THE PROTOCOL SPECIFIED STATISTICAL ANALYSES 
There were no changes from the protocol-specified statistical analyses. 


Statistical Analysis Plan  
 
CONFIDENTIAL 
 
 
Sponsor Reference: 250IPF01 
 
Final: 25 June 2019 
 
Page 17 of 17 
13  
DATA PRESENTATION 
13.1 Insufficient Data for Presentation 
Some of the TFLs may not have sufficient numbers of subjects or data for presentation. If this 
occurs, the blank TFL shell will be presented with a message printed in the center of the table, 
such as, “No serious adverse events occurred for this study.” 
14  
REFERENCES 
1. International Conference on Harmonisation of Technical Requirements for Registration of 
Pharmaceuticals for Human Use, ICH Harmonised Tripartite Guideline, Statistical Principles 
for Clinical Trials (E9), 5 February 1998. 
2. International Conference on Harmonisation of Technical Requirements for Registration of 
Pharmaceuticals for Human Use, ICH Harmonised Tripartite Guideline, Structure and 
Content of Clinical Study Reports (E3), 30 November 1995.