Conditions: Diarrhea-predominant Irritable Bowel Syndrome
Keywords: Irritable Bowel Syndrome diarrhea BOS-589
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 133
Healthy volunteers: no

Brief summary:
This study is being conducted to evaluate in participants with diarrhea-predominant Irritable Bowel Syndrome (IBS-D) the abdominal pain response to BOS-589 after 4 weeks of treatment and to evaluate the overall safety and tolerability of BOS-589 in the treatment of IBS-D during 4 weeks of treatment, relative to placebo (PBO).

Detailed description:

Arms:
arm 1: Participants will receive a high dose of BOS-589 orally twice a day (BID). arm 2: Participants will receive a low dose of BOS-589 orally BID. arm 3: Participants will receive matching placebo orally BID.

Interventions: intervention 1: BOS-589 intervention 2: Placebo
Intervention descriptions: intervention 1: oral tablets intervention 2: oral tablets

Protocol document:
Prot_000.pdf: 
A Phase 2a, Randomized, Double-blind, Placebo-controlled, Multicenter 
Study to Evaluate the Efficacy, Safety, and Tolerability of BOS-589 in the
Treatment of Patients with Diarrhea-predominant Irritable Bowel Syndrome 
(IBS-D) 
ClinicalTrials.gov Identifier: NCT03977155 
Final Protocol, Version 3.0 (Amendment 2), dated 12-August-2019
Final Protocol, Version 2.0 (Amendment 1), dated 0-May-2019
Final Protocol, Version 1.0, dated 25-JanXDU\-2019


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


 
CLINICAL TRIAL PROTOCOL 
Study Title: 
A Phase 2a, Randomized, Double-blind, 
Placebo-controlled, Multicenter Study to Evaluate the 
Efficacy, Safety, and Tolerability of BOS-589 in the 
Treatment of Patients with Diarrhea-predominant Irritable 
Bowel Syndrome (IBS-D) 
Study Number: 
BOS-589-201 
Regulatory 
Identification 
Number(s):  
IND 142421 
Study Phase: 
2a 
Investigational Product: 
BOS-589 
Indication: 
Diarrhea-predominant Irritable Bowel Syndrome 
Sponsor: 
Boston Pharmaceuticals, Inc. 
55 Cambridge Parkway, Suite 400 
Cambridge, Massachusetts 02142 
USA 
 
Version 
Date 
Final v.2.0 
09 May 2019 
Confidentiality Statement 
The information in this document is confidential and will not be disclosed to others 
without written authorization from the sponsor, except to the extent necessary to 
obtain informed consent from persons receiving the study drug or their legal 
guardians, or for discussions with Regulatory Authorities, Institutional Review Boards, 
Independent Ethics Committees, or persons participating in the conduct of the study. 
Do not copy or distribute without written permission from the sponsor. 


Boston Pharmaceuticals, Inc.
BOS-589-201
Protocol
Version 2.0
Confidential
Page 2 of 78
SPONSOR SIGNATURE PAGE
Protocol Title: A Phase 2a, Randomized, Double-blind, Placebo-controlled, Multicenter 
Study to Evaluate the Efficacy, Safety, and Tolerability of BOS-589 in the Treatment of 
Patients with Diarrhea-predominant Irritable Bowel Syndrome (IBS-D)
Protocol Number: BOS-589-201
Version Number and Date: v.2.0, 09 May 2019
I, the undersigned, have approved the final version of this Clinical Trial Protocol.
Date
Date


PPD
PPD


Boston Pharmaceuticals, Inc. 
BOS-589-201 
Protocol 
Version 2.0 
 
Confidential 
 
Page 3 of 78 
INVESTIGATOR AGREEMENT 
Protocol Title: A Phase 2a, Randomized, Double-blind, Placebo-controlled, Multicenter 
Study to Evaluate the Efficacy, Safety, and Tolerability of BOS-589 in the Treatment of 
Patients with Diarrhea-predominant Irritable Bowel Syndrome (IBS-D) 
Protocol Number: BOS-589-201 
Version Number and Date: v.2.0, 09 May 2019 
I, the undersigned, have read the protocol and agree to conduct the trial in compliance 
with the International Conference on/Council for Harmonization (ICH) guidelines and 
any other applicable regulatory requirements; as well as Good Clinical Practice (GCP) 
standards (CPMP/ICH/135/95). 
I will provide copies of the protocol and all pertinent information to all individuals who 
assist in the conduct of this study. I will discuss this material with them to ensure they 
are fully informed regarding the investigational product and the conduct of the study. 
I will use only the Informed Consent Form approved by the sponsor or its representative 
and will fulfill all responsibilities for submitting pertinent information to the Institutional 
Review Board/Independent Ethics Committee (IRB/IEC) responsible for this study. 
I agree that the sponsor or its representatives will have access to any source 
documents from which case report form information may have been generated. 
 
 
 
 
Investigator's Signature 
 
Date 
Name of Investigator (typed or printed) 
Institution Name  
 
 


Boston Pharmaceuticals, Inc. 
BOS-589-201 
Protocol 
Version 2.0 
 
Confidential 
 
Page 4 of 78 
TABLE OF CONTENTS 
TABLE OF CONTENTS .................................................................................................. 4 
LIST OF TABLES ............................................................................................................ 7 
LIST OF FIGURES .......................................................................................................... 7 
LIST OF APPENDICES ................................................................................................... 7 
ABBREVIATIONS ........................................................................................................... 8 
1 
PROTOCOL SUMMARY ........................................................................................ 11 
1.1 
Synopsis ............................................................................................................. 11 
1.2 
Study Schema .................................................................................................... 16 
1.3 
Schedule of Activities ......................................................................................... 17 
2 
INTRODUCTION .................................................................................................... 19 
2.1 
Study Rationale .................................................................................................. 19 
2.2 
Background ........................................................................................................ 19 
2.2.1 
Disease Under Study .................................................................................... 19 
2.2.2 
The “RET” (“REarranged During Transfection”) Gene and the Enteric 
Nervous System ........................................................................................... 20 
2.2.3 
Investigational Product .................................................................................. 21 
2.2.3.1 
Nonclinical Summary ............................................................................... 21 
2.2.3.1.1 
In Vitro Studies ................................................................................... 21 
2.2.3.1.2 
In Vivo Studies ................................................................................... 22 
2.2.3.1.3 
Pharmacokinetics and Product Metabolism in Animals ...................... 22 
2.2.3.1.4 
Toxicology and Nonclinical Assessment of Safety ............................. 23 
2.2.3.2 
Clinical Experience .................................................................................. 24 
2.3 
Risk/Benefit Assessment .................................................................................... 25 
2.3.1 
Risk Assessment ........................................................................................... 26 
2.3.2 
Benefit Assessment ...................................................................................... 26 
2.3.3 
Overall Risk/Benefit Conclusion .................................................................... 26 
3 
OBJECTIVES AND ENDPOINTS ........................................................................... 27 
4 
STUDY DESIGN ..................................................................................................... 28 
4.1 
Overall Design .................................................................................................... 28 
4.1.1 
Study Duration for Patients ........................................................................... 29 
4.1.2 
Number of Patients ....................................................................................... 29 
4.1.3 
Replacement of Patients ............................................................................... 29 
4.1.4 
Number of Sites ............................................................................................ 29 
4.2 
Rationale for Study Design ................................................................................. 29 
4.3 
Justification for Dose .......................................................................................... 30 
4.4 
End of Study Definition ....................................................................................... 31 
5 
STUDY POPULATION ............................................................................................ 32 
5.1 
Inclusion Criteria ................................................................................................. 32 
5.2 
Exclusion Criteria ................................................................................................ 34 


Boston Pharmaceuticals, Inc. 
BOS-589-201 
Protocol 
Version 2.0 
 
Confidential 
 
Page 5 of 78 
5.3 
Lifestyle Considerations ..................................................................................... 38 
5.4 
Screen Failures .................................................................................................. 38 
6 
INVESTIGATIONAL PRODUCT ............................................................................. 39 
6.1 
Investigational Products ..................................................................................... 39 
6.1.1 
Investigational Product Description ............................................................... 39 
6.2 
Preparation/Handling/Storage/Accountability ..................................................... 39 
6.2.1 
Acquisition and accountability ....................................................................... 39 
6.2.2 
Product Storage and Stability ........................................................................ 39 
6.3 
Measures to Minimize Bias ................................................................................. 40 
6.4 
Dosing and Administration .................................................................................. 40 
6.5 
Dose modification ............................................................................................... 41 
6.6 
Investigational Product Compliance ................................................................... 41 
6.7 
Concomitant Therapy ......................................................................................... 41 
6.7.1 
Rescue Medicine ........................................................................................... 42 
6.8 
Intervention After the End of the Study ............................................................... 42 
7 
DISCONTINUATION / WITHDRAWAL ................................................................... 43 
7.1 
Discontinuation of Investigational Product .......................................................... 43 
7.2 
Discontinuation/Withdrawal from the Study ........................................................ 43 
7.3 
Lost to Follow-up ................................................................................................ 45 
8 
STUDY ASSESSMENTS AND PROCEDURES ..................................................... 46 
8.1 
Study Periods ..................................................................................................... 46 
8.2 
Screening Assessments ..................................................................................... 46 
8.3 
Efficacy Assessments ......................................................................................... 47 
8.3.1 
Worst Abdominal Pain ................................................................................... 47 
8.3.2 
Bristol Stool Form Scale ................................................................................ 47 
8.3.3 
IBS-D Global Symptom Score ....................................................................... 47 
8.3.4 
IBS Severity Score System ........................................................................... 48 
8.4 
Safety and Other Assessments .......................................................................... 48 
8.4.1 
Adverse Events and Serious Adverse Events ............................................... 48 
8.4.2 
Physical Examinations .................................................................................. 48 
8.4.3 
Vital Signs ..................................................................................................... 48 
8.4.4 
Electrocardiograms ....................................................................................... 48 
8.4.5 
Clinical Safety Laboratory Assessments ....................................................... 49 
8.5 
Adverse Events and Serious Adverse Events .................................................... 49 
8.5.1 
Definition of Adverse Events ......................................................................... 49 
8.5.2 
Events of Special Interest.............................................................................. 50 
8.5.3 
Definition of Serious Adverse Events ............................................................ 50 
8.5.4 
Classification of an Adverse Event ................................................................ 51 
8.5.4.1 
Assessment of Severity ........................................................................... 51 
8.5.4.2 
Assessment of Causality .......................................................................... 51 
8.5.4.3 
Expectedness .......................................................................................... 52 


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8.5.5 
Time Period and Frequency for Event Assessment and Follow-up ............... 52 
8.5.6 
Adverse Event Reporting .............................................................................. 52 
8.5.7 
Reporting of Pregnancy ................................................................................ 53 
8.6 
Treatment of Overdose ....................................................................................... 54 
8.7 
Pharmacokinetics ............................................................................................... 54 
8.8 
Biomarkers/Pharmacodynamics ......................................................................... 55 
9 
STATISTICAL CONSIDERATIONS ........................................................................ 55 
9.1 
Timing of Analyses ............................................................................................. 55 
9.2 
Sample Size Determination ................................................................................ 55 
9.3 
Populations for Analyses .................................................................................... 56 
9.4 
Statistical Analyses ............................................................................................. 56 
9.4.1 
Efficacy Analyses .......................................................................................... 56 
9.4.1.1 
Primary Endpoint ..................................................................................... 56 
9.4.1.2 
Secondary Endpoint ................................................................................ 57 
9.4.1.3 
Other Efficacy Endpoints ......................................................................... 57 
9.4.1.4 
Missing Data ............................................................................................ 57 
9.4.2 
Safety Analyses ............................................................................................ 58 
9.4.2.1 
Adverse Events ........................................................................................ 58 
9.4.2.2 
Clinical Laboratory Evaluations ................................................................ 58 
9.4.2.3 
Twelve-lead Electrocardiograms, Vital Signs, and Physical 
Examinations ........................................................................................... 58 
9.4.3 
Other Analyses .............................................................................................. 59 
9.4.3.1 
Pharmacokinetic Analysis ........................................................................ 59 
9.4.4 
Baseline Descriptive Statistics ...................................................................... 59 
9.4.5 
Subgroup Analyses ....................................................................................... 59 
9.5 
Planned Interim Analyses ................................................................................... 59 
10 SUPPORTING DOCUMENTATION AND OPERATIONAL 
CONSIDERATIONS................................................................................................ 60 
10.1 Regulatory, Ethical, and Study Oversight Considerations .................................. 60 
10.1.1 
Regulatory and ethical considerations .......................................................... 60 
10.1.2 
Informed Consent/Assent Process ................................................................ 60 
10.1.3 
Confidentiality and Privacy ............................................................................ 61 
10.1.4 
Quality Assurance and Quality Control ......................................................... 62 
10.1.5 
Data Handling and Record Keeping .............................................................. 62 
10.1.5.1 
Data Collection and Management Responsibilities .................................. 62 
10.1.5.2 
Study Records Retention ......................................................................... 63 
10.1.6 
Study and Site Discontinuation and Closure ................................................. 63 
10.1.7 
Dissemination of Clinical Study Data ............................................................ 64 
10.1.8 
Publication Policy .......................................................................................... 64 
11 REFERENCES ....................................................................................................... 65 
12 APPENDICES ......................................................................................................... 68 


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LIST OF TABLES 
Table 1. 
Schedule of Activities ............................................................................... 17 
Table 2. 
Protocol-required Safety Laboratory Assessments .................................. 69 
Table 3. 
Highly Effective Contraceptive Methods .................................................. 73 
 
LIST OF FIGURES 
Figure 1. 
Study Schematic ...................................................................................... 16 
 
LIST OF APPENDICES 
Appendix 1. Clinical Laboratory Tests ......................................................................... 69 
Appendix 2. Contraceptive Guidance and Collection of Pregnancy Information .......... 71 
Appendix 3. The Bristol Stool Form Scale ................................................................... 75 
Appendix 4. Concomitant Medications ......................................................................... 76 
 


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ABBREVIATIONS 
AE 
adverse event 
AKT 
protein kinase B 
ALT 
alanine aminotransferase 
AST 
aspartate aminotransferase 
AUC 
area under the concentration versus time curve 
AUC0-4 
area under the concentration versus time curve from time zero to 
4 hours postdose 
AUC0-12 
area under the concentration versus time curve from time zero to 
12 hours postdose 
AUC0-t 
area under the concentration versus time curve from time zero to 
the last measurable concentration  
bid 
twice daily 
BSFS 
Bristol Stool Form Scale  
BMI 
body mass index 
CFR 
Code of Federal Regulations  
Cmax 
maximum plasma concentration 
Cmin 
minimum plasma concentration 
CNS 
central nervous system 
CONSORT 
Consolidated Standards of Reporting Trials 
CRF 
case report form 
CSR 
clinical study report  
CYP 
cytochrome P450 
ECG 
electrocardiogram 
EDC 
electronic data capture  
ENS 
enteric nervous system 
ERK 
extracellular signal-regulated kinase 
FDA 
Food and Drug Administration 
FDR 
first-degree relative 
GCP 
Good Clinical Practice 
GDNF 
glial cell line-derived neurotrophic factor 
GFR-α 
glial cell line-derived neurotrophic factor family receptor-alpha 
GI 
gastrointestinal 
GLP-1 
glucagon-like peptide-1  
HBV 
hepatitis B virus 
HCV 
hepatitis C virus 


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HDPE 
high-density polyethylene 
hERG 
human ether-à-go-go related gene  
HIV 
human immunodeficiency virus 
HRT 
hormone replacement therapy 
IB 
Investigator’s Brochure 
IBS 
irritable bowel syndrome 
IBS-C 
irritable bowel syndrome-constipation 
IBS-GS 
Irritable Bowel Syndrome Global Scale 
IBS-D 
irritable bowel syndrome-diarrhea 
IBS-M 
irritable bowel syndrome-mixed 
IBS-SS 
Irritable Bowel Syndrome Severity Score 
IBS-U 
irritable bowel syndrome-unclassified 
IC50 
half-maximal inhibitory concentration 
ICF 
Informed Consent Form 
ICH 
International Conference on/Council for Harmonisation  
IEC 
Independent Ethics Committee 
ILC3 
group 3 innate lymphoid cells  
IRB 
Institutional Review Board 
ITT 
intention-to-treat 
IV 
intravenous 
IWRS 
interactive web response system  
JAK 
Janus kinase 
MedDRA 
Medical Dictionary for Regulatory Activities 
NOAEL 
no observed adverse effect level 
NRS 
numeric rating scale 
NTEAE 
non-treatment-emergent adverse event  
PBO 
placebo 
PD 
pharmacodynamic(s) 
PE 
physical examination 
P-gp 
P-glycoprotein  
PI3K 
phosphoinositide 3-kinase  
PK 
pharmacokinetic(s) 
PRO 
patient-reported outcome 
PYY 
pancreatic peptide YY 
QC 
quality control 
QTcB 
QT interval corrected by Bazett’s formula  


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QTcF 
QT interval corrected by Fridericia’s formula 
RAS 
rat sarcoma  
RBC 
red blood cell 
RET 
REarranged during Transfection 
SAE 
serious adverse event 
SAP 
Statistical Analysis Plan 
SC 
subcutaneous 
SD 
standard deviation 
SK-N-AS 
a human neuroblastoma cell line 
STAT 
signal transducer and activator of transcription proteins 
SUSAR 
suspected unexpected serious adverse reaction 
TEAE 
treatment-emergent adverse event 
Tmax 
time to maximum concentration 
TT 
a human thyroid medullary carcinoma cell line 
ULN 
upper limit of normal 
WAP 
worst abdominal pain 
WOCBP 
woman of childbearing potential  
 


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1 
PROTOCOL SUMMARY 
1.1 
Synopsis  
Sponsor: 
Boston Pharmaceuticals Inc.  
Title: 
A Phase 2a, Randomized, Double-blind, Placebo-controlled, 
Multicenter Study to Evaluate the Efficacy, Safety, and 
Tolerability of BOS-589 in the Treatment of Patients with 
Diarrhea-predominant Irritable Bowel Syndrome (IBS-D) 
Protocol Number: 
BOS-589-201 
Phase: 
2a 
Objectives and 
Endpoints: 
OBJECTIVES 
ENDPOINTS 
Primary 
x To evaluate in patients 
with IBS-D the abdominal 
pain response to BOS-
589 after 4 weeks of 
treatment, relative to 
placebo (PBO). 
x 24-hour worst abdominal 
pain scores (WAP) at 
Day 29 compared to 
baseline (averaged over 
the week prior to each 
respective timepoint). 
x To evaluate the overall 
safety and tolerability of 
BOS-589 in the treatment 
of IBS-D during 4 weeks 
of treatment, relative to 
PBO. 
x Incidence of adverse 
events (AEs), serious 
adverse events (SAEs), 
discontinuations because 
of AEs, and any 
treatment-related severe 
AEs. 
Secondary 
x To evaluate the treatment 
effect of BOS-589 on 
defecation after 4 weeks, 
relative to PBO. 
x Change in stool 
consistency, measured by 
the daily Bristol Stool 
Form Score (BSFS) at 
Day 29 compared to 
baseline (averaged over 
the week prior to each 
respective timepoint). 
x 
Change in stool 
frequency, measured by 
the total number of  


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spontaneous bowel 
movements in 24 hours at 
Day 29 compared to 
baseline (averaged over 
the week prior to each 
respective timepoint). 
x To evaluate the treatment 
effect of BOS-589 on 
IBS-related signs and 
symptoms. 
x Change in the IBS 
Severity Score (IBS-SS) 
at Day 29 compared to 
baseline. 
x Change in the IBS Global 
Scale (IBS-GS) at Day 29 
compared to baseline 
(averaged over the week 
prior to each respective 
timepoint). 
x To evaluate the 
steady-state 
pharmacokinetics (PK) of 
BOS-589. 
x Maximum observed 
plasma concentration 
(Cmax); time to Cmax (Tmax); 
minimum plasma 
concentration (Cmin); area 
under the concentration 
versus time curve (AUC) 
from time zero to 4 hours 
postdose (AUC0-4); AUC 
from time zero to the last 
quantifiable concentration 
(AUC0-t). 
Study Design: 
A phase 2a, randomized, double-blind, PBO-controlled, 
multicenter trial to provide proof-of-principle efficacy of 
BOS-589 in IBS-D patients and to inform dose selection for 
subsequent development.  
The study will comprise 3 phases:  
A Pre-treatment Phase of up to 4 weeks, in which all 
patients will be assessed to determine eligibility. This phase 
will consist of initial screening assessments after which 
eligible patients will enter a Run-in Period of up to 3 weeks.   
During the Run-in period the patients will complete an 
electronic diary to collect daily information related to their 
IBS-D symptoms, bowel function, and rescue medication use, 
to confirm disease activity, and diary compliance.  


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Upon completion of the Run-in Period, patients will return to 
the study site to confirm eligibility for randomization into the 
double-blinded Treatment Phase. 
A Treatment Phase, in which eligible patients will be 
randomized (1:1:1) to receive BOS-589 or PBO twice daily 
(bid) for a total of 4 weeks. BOS-589 will be administered at 
1 of 2 dose levels. Patients will be randomized to the 
following cohorts:   
x Cohort 1 (High Dose): 200 mg BOS-589 bid orally;  
x Cohort 2 (Low Dose): 50 mg BOS-589 bid orally; 
x Cohort 3 (PBO): matching PBO oral tablets bid. 
During the treatment phase, patients will continue to 
complete the electronic diary to collect daily information 
related to their IBS-D symptoms, bowel function, and rescue 
medicine. 
A Post-treatment Phase, in which all patients who complete 
4 weeks of treatment will return to the clinical for a 2-week 
follow-up visit.   
Patients who discontinue treatment early will be asked to 
return to the clinic for safety assessments.       
Study Population: 
Male and female patients 18 to 65 years of age, inclusive, 
with a diagnosis of IBS-D. 
Number of Patients 
Planned: 
Approximately 300 patients will be screened with the intent of 
randomizing 132 patients for the study. 
Duration of Patient 
Participation and 
Study: 
The duration of patient participation is anticipated to be up to 
10 weeks. 
The duration of the study is anticipated to be approximately 
12 months. 
Study Sites: 
Up to 66 sites in the United States. 
Investigational 
Product: 
BOS-589 (oral doses bid). 
Reference 
Treatment: 
Matching PBO (oral doses bid). 
Concomitant 
Product: 
Not applicable. 


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Statistical Methods: 
Efficacy: 
This is a phase 2a proof-of-principle study. Descriptive and 
inferential statistical methods will be employed. A standalone 
Statistical Analysis Plan will be developed and approved prior 
to database lock. 
A hierarchical hypothesis test will be employed for the 
primary efficacy endpoint, defined as a change from baseline 
to Day 29 in the patient-reported outcome for WAP measured 
daily on an 11-point numeric rating scale. Mean change from 
baseline will be measured as a continuous variable and 
compared between treatment groups in the following testing 
order using a t-test of equal variances with a two-tailed alpha 
of 0.05: 
Hypothesis 1: 
Active Treatment Group (Cohort 1 + Cohort 2) versus 
PBO (Cohort 3); 
If statistically significant at P < 0.05, then proceed to 
Hypothesis 2. 
Hypothesis 2: 
Cohort 1 (High Dose) versus Cohort 3 (PBO); 
If statistically significant at P < 0.05, then proceed to 
Hypothesis 3. 
Hypothesis 3: 
Cohort 2 (Low Dose) versus Cohort 3 (PBO); 
If statistically significant at P < 0.05 then, proceed to 
Hypothesis 4. 
Hypothesis 4: 
Cohort 1 (High Dose) versus Cohort 2 (Low Dose). 
Safety: 
Adverse events will be recorded from the time of consent until 
the Day 43/End-of-Study Follow-up visit. The number and 
percentage of patients with AEs will be displayed by system 
organ class and preferred term using Medical Dictionary for 


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Regulatory Activities Version 20.0 or higher, by study 
treatment. Summaries in terms of severity and relationship to 
investigational product will also be provided. All serious AEs 
will be summarized in a similar manner. Patient listings of all 
AEs causing discontinuation of investigational product and all 
SAEs will be produced. 
All AEs will be listed for individual patients, showing both 
verbatim and preferred terms. Separate summaries of 
treatment-emergent SAEs and treatment-emergent AEs 
related to investigational product will be generated. 


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1.2 
Study Schema 
Figure 1. Study Schematic 
 


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1.3 
Schedule of Activities 
Table 1. 
Schedule of Activities 
 
Screening 
Days -29 to -1, 
Study Visit 1 
Run-in Perioda 
Days -15 to -1, 
Study Visit 2 
Baseline 
Day 1, 
Study Visit 3 
Day 8 (± 1 day) 
Study Visit 4 
 
Day 15 (± 1 day) 
Study Visit 5 
 
Day 22 (± 1 day) 
Study Visit 6 
 
Day 29 (± 1 day) 
End of Treatment 
/Early Terminationb 
Study Visit 7  
Day 43 (± 3 days) 
End of Study 
Study Visit 8 
 
Procedures 
Pre-treatment 
Treatment 
Follow-up 
Informed consent 
X 
 
 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
Medical and surgical history 
X 
 
 
 
 
 
 
 
Rome IV assessment 
X 
 
 
 
 
 
 
 
Concomitant medication review 
X 
X 
X 
X 
X 
X 
X 
X 
Adverse event review and evaluation 
 
X- - - - - - - - - - - - - - - - - - - - - - - - - - - - continuous - - - - - - - - - - - - - - - - - - - - - - - - - - - X 
Physical examinationc  
X 
 
X 
 
 
 
X 
 
Height, weight, and body mass index 
Xd 
 
X 
 
X 
 
X 
X 
Vital signs 
X 
 
X 
X 
X 
X 
X 
X 
12-lead electrocardiogram 
X 
 
 
 
 
 
X 
 
Clinical safety laboratory testse 
X 
 
X 
 
X 
 
X 
X 
HIV, HCV/HBV testing 
X 
 
 
 
 
 
 
 
Urine pregnancy test  
X 
 
X 
 
 
 
X 
X 
C-reactive protein and erythrocyte 
sedimentation rate tests 
X 
 
X 
 
X 
 
X 
X 
Biomarker sample collection 
 
 
X 
 
X 
 
X 
X 
Pharmacokinetic sample collectionf 
 
 
X 
X 
X 
X 
 
 
Worst abdominal pain 
X 
Xg 
X- - - - - - - - - - - - - - - - - - - - - - -h- - - - - - - - - - - - - - - - - - - - - X 
Xh 
Bristol Stool Form Scale 
X 
Xg 
X- - - - - - - - - - - - - - - - - - - - - - -h- - - - - - - - - - - - - - - - - - - - - X 
Xh 


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Screening 
Days -29 to -1, 
Study Visit 1 
Run-in Perioda 
Days -15 to -1, 
Study Visit 2 
Baseline 
Day 1, 
Study Visit 3 
Day 8 (± 1 day) 
Study Visit 4 
 
Day 15 (± 1 day) 
Study Visit 5 
 
Day 22 (± 1 day) 
Study Visit 6 
 
Day 29 (± 1 day) 
End of Treatment 
/Early Terminationb 
Study Visit 7  
Day 43 (± 3 days) 
End of Study 
Study Visit 8 
 
Procedures 
Pre-treatment 
Treatment 
Follow-up 
Spontaneous bowel movements (total 
number of stools) 
 
Xg 
X- - - - - - - - - - - - - - - - - - - - - - -h- - - - - - - - - - - - - - - - - - - - - X 
Xh 
IBS-GS 
 
Xg 
X- - - - - - - - - - - - - - - - - - - - - - -h- - - - - - - - - - - - - - - - - - - - - X 
Xh 
Rescue medications 
 
Xg 
X- - - - - - - - - - - - - - - - - - - - - - -h- - - - - - - - - - - - - - - - - - - - - X 
Xh 
IBS-SS 
 
 
X 
 
 
 
X 
X 
Eligibility review 
X 
 
X 
 
 
 
 
 
Randomization 
 
 
X 
 
 
 
 
 
Administration of investigational product 
 
 
Day 1 - - - - - - - - - - - - - - - - - - - - - - - - - - - Day 28 
 
 
HBV = hepatitis B virus; HCV = hepatitis C virus; HIV = human immunodeficiency virus; IBS-GS = Irritable Bowel Syndrome Global Scale; 
IBS-SS = Irritable Bowel Syndrome Severity Score.  
a 
Run-in may be extended by up to one week in accordance with the investigators judgement. 
b 
Procedures performed at Visit 7/End of Treatment or upon Early Termination. Early Termination procedures are to be performed in close 
proximity to discontinuation of dosing for patients who are withdrawn from the study. Other procedures may be performed at Investigator or 
Sponsor discretion. 
c 
Full physical exam to be conducted at Screening. Symptom-directed physical examinations should be conducted at subsequent visits. 
d 
Height and body mass index to be collected at Screening Visit only. 
e 
Hematologic, complete serum chemistry, urinalysis panels and fecal calprotectin to be completed as noted in Appendix 1. 
f 
Pharmacokinetic (PK) serial blood samples are to be collected on Day 1 and Day 15 at predose and at 0.5, 1, 2, and 4 hours postdose. If PK 
serial blood samples cannot be collected on Day 15, PK serial blood samples should be collected on Day 22. A predose sample will be 
obtained on all other Treatment Study Visits. Allowable sampling windows for PK blood draws will be 30 minutes prior to dosing for the predose 
sample, then ± 5 minutes for 0.5 and 1 hour postdose, and ± 10 minutes for 2 and 4 hours postdose.  
g 
Data will be collected daily for entire Run-in Period. 
h 
Data will be collected daily until Early Termination or until End-of-Study Visit, whichever is later. 
 


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2
INTRODUCTION
2.1
Study Rationale
Irritable bowel syndrome (IBS) is a gastrointestinal (GI) illness with a prevalence of 
approximately 5% to 20% globally and characterized by a constellation of clinical 
symptoms. Establishing a diagnosis and assessing response to treatment remains 
challenging because there are no biomarkers that reliably correlate with disease state.
BOS-589 is an oral agent with limited systemic absorption that inhibits RET 
(REarranged during Transfection), a receptor tyrosine kinase that is hypothesized to 
play a key role in the maintenance of a healthy enteric nervous system (ENS). Inhibition
of RET may represent a novel therapeutic strategy for the treatment of IBS by 
attenuating visceral hypersensitivity and/or colonic motility.
BOS-589 has been evaluated in healthy human subjects at doses up to 200 mg twice 
daily (bid) for 2 weeks. Consistent with its low oral bioavailability, BOS-589 was safe 
and well tolerated with no significant safety signals identified. The purpose of this study 
is to evaluate the efficacy, safety, and tolerability of BOS-589 in the treatment of 
patients with diarrhea-predominant IBS (IBS-D).
2.2
Background 
2.2.1
Disease Under Study
Irritable bowel syndrome is a relatively common GI illness characterized by a number of 
clinical symptoms, including abdominal pain and discomfort, abnormal bowel habits, 
and bloating (
). Subtypes of IBS, based on the 
predominant bowel habit(s) reported by IBS patients, include diarrhea (IBS-D), 
constipation (IBS-C), mixed (IBS-M), or unclassified (IBS-U). Currently approved 
medications for IBS address the restoration of patients’ bowel habits and are minimally 
effective in addressing abdominal pain and discomfort. Because the etiology of the 
disease has not been clearly established, diagnosis is difficult and relies primarily on the 
presence of a specific symptom complex occurring in the absence of an alternative 
explanation. The development of criteria by expert panels, with recent iterations as 
recently as 2016, has improved the diagnosis and management of IBS patients 
(
). 
It is generally believed that the sensory inputs/outputs in the ENS and central nervous 
system (CNS) are altered in patients with IBS and this contributes to the signs and 
symptoms they experience. For example, patients with IBS have a heightened and 
disproportionate sensory experience (visceral hypersensitivity) for a given stimulus 
(
). Visceral hypersensitivity and abnormal bowel habits may result 
from visceral afferent neurons or increased nerve fiber density and sprouting that have 
been observed in the intestinal mucosal tissues of IBS patients (
; 
). The sensitizing event causing visceral 
hypersensitivity may be transient or chronic; however, its impact on the CNS and ENS 
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may be long lasting (
). Supernatants from these intestinal 
tissues can stimulate nerve outgrowth and increase neuronal signaling in vitro, 
suggesting that neurotrophic factors and/or inflammatory cytokines might be mediators 
of neuronal plasticity in IBS (
; 
). Given the central role the ENS may play in mediating signals 
between the CNS and the intestine, ENS modulation may provide therapeutic benefit to 
IBS patients by impacting visceral hypersensitivity and intestinal motility.
The only drugs currently approved by the Food and Drug Administration (FDA) for the 
treatment of IBS are alosetron (for women only), rifaximin, and eluxadoline for refractory 
IBS-D, and plecanatide, linaclotide, and lubiprostone for IBS-C. The majority of these 
drugs, and those used off label to treat IBS, target patients’ symptoms by altering GI 
motility and are minimally effective in addressing abdominal pain and discomfort. Of the 
FDA-approved agents, only alosetron is hypothesized to modulate the CNS and ENS 
(
). Therefore, an agent such as BOS-589, 
which may ameliorate visceral hypersensitivity and potentially impact intestinal motility, 
would address a significant unmet medical need for IBS-associated pain.
2.2.2
The “RET” (“REarranged During Transfection”) Gene and the Enteric 
Nervous System
The RET gene, localized on human chromosome 10q11.2, encodes a receptor-type 
tyrosine kinase with an extracellular domain, a transmembrane domain, and an 
intracellular tyrosine kinase domain (
). The ligands for RET have 
been identified as neurotrophic factors of the glial cell line-derived neurotrophic factor 
(GDNF) family, including GDNF, neurturin, artemin, and persephin. Ligand binding to its 
corresponding GDNF family receptor-alpha (GFRα) co-receptor triggers RET 
dimerization and subsequent transphosphorylation of intracellular tyrosines 
(
) and leads to the activation of different intracellular signaling 
cascades, including the Janus kinase/signal transducer and activator of transcription 
proteins (JAK/STAT), phosphoinositide 3-kinase/protein kinase B (PI3K/AKT), and rat 
sarcoma/extracellular signal-regulated kinase (RAS/ERK) pathways.
Mice deficient in the GDNF ligand, its coreceptor GFRα1, or the RET protein itself, 
exhibit severe defects in kidney and ENS development. This implicates RET signaling 
as critical to the development of normal kidneys and the ENS (
). The 
role of RET in the development of the ENS is also apparent in patients with 
Hirschsprung’s disease, who frequently suffer from colonic obstruction because of a 
lack of normal colonic innervation. In Hirschprung’s disease, different loss-of-function 
mutations that occur in the RET gene account for the highest proportion of both familial 
and sporadic cases of the disease (
). 
While its role during the development of the ENS has been well established, recent 
reports also implicate a significant role for RET in the maintenance and plasticity of the 
adult ENS. Neurons within the submucosal and myenteric plexus of the adult human 
colon have been shown to express RET and its coreceptors, GFRα1 and GFRα2, while 
the GDNF ligand is expressed in the muscularis mucosa and in circular and longitudinal 
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muscle tissue (
). Systemic administration of GDNF in adult 
rodents results in significant increases in submucosal neuron density in both the small 
intestine and colon and altered function (
). Furthermore, a conditional 
knockout of the RET co-receptor, GFRα3, results in decreased colonic hypersensitivity 
implicating a role for RET signaling in visceral nociception (
). 
Therefore, by reducing RET signaling, inhibition of RET may modulate ENS activity.
2.2.3
Investigational Product
BOS-589, formerly GSK3352589, is a potent and selective inhibitor of RET that has 
been shown to reduce visceral hypersensitivity in an animal model of IBS and inhibit 
cholinergic-induced increases in colonic motility (details are provided in the BOS-589 
Investigator’s Brochure [IB]). The results from these preclinical studies suggest that 
inhibition of RET with a potent, selective, and gut-restricted small molecule may 
represent a novel therapeutic strategy for the treatment of abdominal pain and 
defecation abnormalities (i.e., diarrhea) in patients with IBS through the attenuation of 
visceral hypersensitivity and/or cholinergically mediated ion transport and colonic 
motility. The patient population likely to derive the greatest benefit would comprise 
individuals with IBS-D with increased GI motility.
2.2.3.1
Nonclinical Summary
A range of in vitro and in vivo studies have been conducted to investigate the primary, 
secondary, and safety pharmacology of BOS-589. Details are provided in the BOS-589 
IB.
2.2.3.1.1
In Vitro Studies
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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2.2.3.1.2
In Vivo Studies
 
 
 
 
 
 
 
 
 
 
2.2.3.1.3
Pharmacokinetics and Product Metabolism in Animals
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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2.2.3.1.4
Toxicology and Nonclinical Assessment of Safety
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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2.2.3.2
Clinical Experience 
Study 
 evaluated the safety, tolerability, pharmacokinetics (PK), and 
exploratory pharmacodynamics (PD) of escalating single and multiple oral doses of 
BOS-589 ranging from 2 to 400 mg in the fasted state, and 25 mg in the fed state. All 
doses in the single ascending-dose portion of the study were generally safe and well 
tolerated, with a safety profile similar to that of placebo (PBO). Similarly, repeat-dose 
administration of BOS-589 for 14 days at doses ranging from 5 to 200 mg bid in the fed 
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state was generally safe and well tolerated. There were no drug-related clinically 
significant changes in safety laboratory tests, vital signs, ECGs, or stool patterns as 
assessed by the Bristol Stool Form Scale (BSFS) in this study. There were no serious 
adverse events (SAEs) considered related to the administration of BOS-589. A review of 
all GI adverse events (AEs) demonstrated that the occurrence of GI AEs was similar in 
the PBO- and BOS-589-treatment groups. There was no pattern or trend with dose 
escalation suggestive of a treatment effect. 
The PK profile of BOS-589 demonstrated limited oral bioavailability and systemic 
exposure, with plasma concentrations generally less than 2 ng/mL, when measured 
utilizing a sensitive analytical method to detect concentrations as low as 5 pg/mL. 
Exposures were dose dependent and less than dose proportional, although box plots of 
dose-normalized parameters suggested dose proportionality between 15 mg and 
100 mg. Dosing was escalated to the highest possible dose because predicted mean 
systemic exposures did not exceed the defined plasma toxicokinetic limits (AUC0-t and 
Cmax of 40.4 ng*h/mL and 26.7 ng/mL, respectively) and there was no evidence of 
dose-limiting toxicities. There was accumulation of BOS-589 with repeat dosing, but 
systemic exposures remained very low after 14 days of dosing, with respective 
geometric mean (CV%) AUC0-t and Cmax values of 24.1 (45.9) h*ng/mL and 
1.53 (43.6) ng/mL for the highest dose of 200 mg bid; BOS-589 was likely at steady 
state before that time. Accumulation was lower for doses below 100 mg bid (1.3 to 
1.8 fold) and highest for the highest dose of 200 mg bid (2.0 to 2.7 fold).
The study included a pilot food-effect group to evaluate the magnitude of a food 
challenge on the bioavailability of single-dose BOS-589. Following administration of a 
single dose of BOS-589 25 mg in the fed state, there was a small decrease in exposure 
with food, with decreases in mean Cmax and AUC values in the range of 20% to 35%. 
The time to Cmax (Tmax) remained the same with or without food. These food-effect 
results were deemed not clinically important; dosing BOS-589 with or without food is not 
anticipated to affect future evaluation of safety or efficacy.
Because RET is expressed in enteroendocrine cells lining the intestinal mucosa, an 
exploratory objective of the study was to explore the effect of BOS-589 on glucagon-like 
peptide-1 (GLP-1) and pancreatic peptide YY (PYY) excursions in plasma; however, no 
clear relationship or impact on BOS-589 administration and peptide secretion was 
identified.  
2.3
Risk/Benefit Assessment  
Summaries of findings from nonclinical and clinical studies conducted with BOS-589 
can be found in the IB (refer to the IB for additional details). The following section 
outlines the risk assessment and mitigation strategy for this protocol. 
The current study, BOS-589-201, represents the first administration of BOS-589 to 
patients with IBS-D. Considerations for safety monitoring are derived primarily from the 
literature regarding RET expression in the intestine, nonclinical data, and clinical 
experience dosing BOS-589 to normal healthy volunteers (Study
), in 
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which no clinically relevant risks were identified that would preclude dosing a RET 
inhibitor for up to 4 weeks in patients with IBS-D.
2.3.1
Risk Assessment
 
 
 
 
 
 
 
 
 
 
 
 
   
 
 
 
 
 
 
 
 
 
2.3.2
Benefit Assessment
Patients randomized to the active treatment arms may potentially experience 
improvement in their IBS-D during the course of the study. Those randomized to the 
PBO arm are not expected to obtain any benefit beyond that of their background 
treatment. 
2.3.3
Overall Risk/Benefit Conclusion
On the basis of nonclinical and clinical study results to date, limited effective alternative 
treatments, and the strength of the scientific hypothesis under evaluation, BOS-589 is 
considered to have a favorable benefit-risk profile for patients with IBS-D.
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3 
OBJECTIVES AND ENDPOINTS 
OBJECTIVES 
ENDPOINTS 
Primary 
x To evaluate in patients with IBS-D 
the abdominal pain response to 
BOS-589 after 4 weeks of treatment, 
relative to placebo (PBO). 
x 24-hour worst abdominal pain scores 
(WAP) at Day 29 compared to baseline 
(averaged over the week prior to each 
respective timepoint). 
x To evaluate the overall safety and 
tolerability of BOS-589 in the 
treatment of IBS-D during 4 weeks 
of treatment, relative to PBO. 
x Incidence of adverse events (AEs) 
serious adverse events (SAEs), 
discontinuations because of AEs, and 
any treatment-related severe AEs. 
Secondary 
x To evaluate the treatment effect of 
BOS-589 on defecation after 
4 weeks, relative to PBO. 
 
x Change in stool consistency, measured 
by the daily Bristol Stool Form Score 
(BSFS) at Day 29 compared to baseline 
(averaged over the week prior to each 
respective timepoint). 
x 
Change in stool frequency, measured 
by the total number of spontaneous 
bowel movements in 24 hours at Day 29 
compared to baseline (averaged over 
the week prior to each respective 
timepoint). 
x To evaluate the treatment effect of 
BOS-589 on IBS-related signs and 
symptoms. 
 
x Change in the IBS Severity Score 
(IBS-SS) at Day 29 compared to 
baseline. 
x Change in the IBS Global Scale 
(IBS-GS) at Day 29 compared to 
baseline (averaged over the week prior 
to each respective timepoint). 
x 
To evaluate the steady-state 
pharmacokinetics of BOS-589. 
x 
Maximum observed plasma 
concentration (Cmax); time to Cmax (Tmax); 
minimum plasma concentration (Cmin); 
area under the concentration versus 
time curve (AUC) from time zero to 
4 hours postdose (AUC0-4); AUC from 
time zero to the last quantifiable 
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4 
STUDY DESIGN  
4.1 
Overall Design 
This study is a phase 2a, randomized, double-blind, PBO-controlled, multicenter trial 
designed to provide the first proof-of-principle efficacy of BOS-589 in IBS-D patients, 
and to inform dose selection for subsequent development. The study will consist of a 
pre-treatment phase, a 4-week double-blind treatment phase, and a 2-week 
post-treatment follow-up period.  
Pre-Treatment Phase: During the pre-treatment phase, patients will be evaluated for 
up to 4 weeks to assess eligibility. The pre-treatment phase will consist of initial 
screening assessments and a Run-in period. 
After the initial screening assessments have been performed, eligible patients will enter 
a Run-in period of up to 3-weeks. During the Run-in period, the patients will complete 
an electronic diary to collect daily information related to their IBS-D symptoms, bowel 
function, and rescue medicine use, to confirm disease activity and diary compliance. 
Patients will also be requested to discontinue any prohibited medications during this 
phase of the study.  
Upon completion of the Run-in period, patients will return to the study site to confirm 
eligibility for randomization into a 4-week double-blinded Treatment Phase. 
Treatment Phase: Eligible patients will be randomized (1:1:1) into the following cohorts:   
x Cohort 1 (High Dose): 200 mg BOS-589 bid orally  
x Cohort 2 (Low Dose): 50 mg BOS-589 bid orally 
x Cohort 3 (PBO): matching PBO oral tablets bid orally 
During the 4 weeks of double-blind treatment, patients will continue to record their daily 
IBS-D symptoms, bowel function, and rescue medicine use in the electronic diary, as 
described in Section 8.3. 
Post-treatment Phase: A 2-week post-treatment follow-up visit will occur for patients 
who complete the Treatment Phase. During the 2 weeks of follow-up, patients should 
continue to record their daily IBS-D symptoms, bowel function, and rescue medicine use 
in the electronic diary.    
Patients who prematurely discontinue treatment should return to the study center to 
complete the early termination assessments as soon as possible after stopping the 
treatment. 
Data analyses will occur after all patients in the trial have completed the last visit or 
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Post-study Access to Therapy: No post-treatment access to therapy will be provided 
to patients randomized in the study.
4.1.1
Study Duration for Patients
Unless shortened by intolerable AEs or rapid disease progression, each patient’s 
participation in this study is anticipated to last up to 10 weeks. 
The total duration of this study is anticipated to be approximately 12 months, including 
patient enrollment, treatment, and follow-up.
4.1.2
Number of Patients
Approximately 300 patients will be screened to randomize approximately 132 patients 
with IBS-D (44 patients per cohort).
4.1.3
Replacement of Patients
Patients who sign the ICF and are randomized but do not receive the investigational 
product may be replaced. Once randomized, patients who have received at least 1 dose 
of investigational product and are withdrawn from therapy before completing 28 days of 
dosing or are discontinued from the study for any reason will not be replaced. 
4.1.4
Number of Sites
Up to 66 sites in the United States may participate in this study.
4.2
Rationale for Study Design
Key aspects of the study (e.g., eligibility, cohort size, stopping criteria, safety data 
collection, efficacy assessments, use of PBO as control, and use of patient-reported 
outcome [PRO] tools) are based upon generally accepted clinical trial methodologies for 
phase 2a efficacy studies and prior studies conducted in patients with IBS. 
The Rome IV (
) diagnostic criteria for IBS is the best accepted tool for 
standardized IBS diagnosis.
A Run-in Phase is designed to account for fluctuations in symptoms and the potential for 
wide variations in bowel habits; patient diaries that record frequency and severity of 
daily symptoms will be used to ensure that symptom severity fluctuations are identified 
and taken into account for subject eligibility. Patients will also be assessed on their 
ability to record their disease-related information in the required manner.
The use of PBO as a control comparator in IBS clinical trials is acceptable given the 
lack of consistently effective treatments. Placebo is an important component of IBS 
clinical trials given the high PBO effect in this population.  
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The key primary and secondary endpoints in this study are based on PRO 
assessments. These assessments evaluate pain, defecation, and IBS signs and 
symptoms. Abdominal pain intensity is measured using an 11-point (i.e., 0 to 10) 
numeric rating scale (NRS) that asks patients daily to rate their worst abdominal pain 
over the past 24-hours as recommended by the FDA guidance on IBS (FDA, 
May 2012). The use of the NRS for pain has been validated in IBS clinical studies 
(
). The IBS Severity Score (IBS-SS) (
) and IBS 
Global Scale (IBS-GS) (
) are validated and standard methods of 
assessing IBS symptoms in clinical trials. 
Although treatment durations longer than 4 weeks will be required for true assessment 
of efficacy in IBS patients, in this initial phase 2a study, 4 weeks should be sufficient to 
identify any efficacy signal that warrants further clinical invention.  
The use of multiple enrolling sites is aimed at maximizing external validity and to 
minimizing the potential influence of regional variations in diet, exercise habits, and 
ethnicity.
4.3
Justification for Dose
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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4.4
End of Study Definition
The end of the study is defined as completion of the last visit or procedure shown in the 
Schedule of Activities in the trial globally. A patient will be considered to have completed 
the study if he or she has completed all phases of the study, including the last visit or 
the last scheduled procedure shown in the Schedule of Activities, Section 1.3.
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5
STUDY POPULATION
Before any study-specific activities/procedures, the appropriate written informed 
consent must be obtained. Prospective approval of protocol deviations to recruitment 
and enrollment criteria, also known as protocol waivers or exemptions, is not permitted. 
5.1
Inclusion Criteria
In order to be eligible to participate in this study, an individual must meet all of the 
following criteria:
Age
1. Male and female patients must be 18 to 65 years of age, inclusive, at the time of 
signing informed consent.
Run-in eDiary Compliance
2. Patient has completed the daily electronic diary on at least 6 of the 7 days during the 
week prior to randomization AND at least 11 of the 14 days during the 2 weeks prior 
to randomization. Patients have up to 3 weeks to meet these criteria.
Type of Patient and Disease Characteristics
3. Patient meets the diagnosis of IBS based on the Rome IV diagnostic criteria 
(
):*
x
Recurrent abdominal pain occurring, on average, at least 1 day per week and 
associated with 2 or more of the following:
o
Related to defecation; 
o
Associated with a change in frequency of bowel movements;
o
Associated with a change in form (appearance) of stool;
* These criteria must be fulfilled for the last 3 months prior to randomization and 
onset must have occurred at least 6 months prior to randomization. 
4. Patient meets the diagnosis of IBS-D subtype based on Rome IV diagnostic criteria
(
). On days when the patient experiences IBS symptoms:
x
At least 25% of stools are loose or watery; AND 
x
Fewer than 25% of stools are hard.
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5. Over the week prior to randomization, the patient has an average of worst abdominal 
pain (WAP) scores in the prior 24 hours of 4.0 to 8.0 (on a 0 to 10 numerical rating 
scale, where 0 indicates no pain and 10 indicates worst pain imaginable).    
6. Over the week prior to randomization, the patient has an average daily BSFS score 
≥ 5.0 (on a 1 to 7 scale, where 1 = hard, lumpy stools, and 7 = watery, liquid stools) 
and at least 5 days with a BSFS score ≥ 5.0. 
7. Over the week prior to randomization, the patient has an average daily IBS-GS 
score of ≥ 2.0 (on a 0 to 4 scale, where 0 = none, 1 = mild, 2 = moderate, 3 = severe, 
4 = very severe). 
8. Patient is not planning to change his or her usual diet and lifestyle during the course 
of the study.  
Diagnostic Assessments 
9. Patient must undergo or previously have undergone (a) an appropriate evaluation for 
their IBS symptoms, including an evaluation for organic/structural etiologies (if in the 
presence of alarm symptoms); and (b) age-appropriate screening for colorectal 
cancer, if applicable. 
a) If at least one of the following alarm features are present, then the patient must 
have had a colonoscopy since the onset of symptoms or within the past 5 years 
(whichever is less):  
x Documented and unexplained weight loss of ≥ 10% within the past 
6 months; 
x Nocturnal diarrhea; 
x Blood mixed with stool (except hemorrhoidal bleeding, defined as 
occasional blood found on the toilet paper only or limited dripping of blood 
into the toilet bowl after defecation; 
x Unexplained iron-deficiency anemia. 
b) If no alarm features are present, then the patient must have had a colonoscopy 
or other appropriate exam, based on criteria as outlined below: 
x Age ≥ 50 (≥ 45 if African-American): Colonoscopy within the past 10 years; 
x First-degree relative (FDR) diagnosed with colorectal cancer under age 60 
OR 2 FDRs diagnosed with colorectal cancer at any age: Colonoscopy 
within past 5 years, beginning 10 years before age of youngest FDR (at 
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x Age ≥ 40 AND FDR diagnosed with colorectal cancer (at any age): 
Colonoscopy within past 10 years; 
10. Patient is negative for serum tissue transglutaminase IgA antibody (tTG-IgA) plus 
has evidence of detectable serum IgA within the normal reference range.   
Weight  
11. Body mass index (BMI) within the range 16 to 39 kg/m2 (inclusive).  
Gender  
12.  Male and female patients are eligible if:  
a. Male patients: 
A male patient must agree to use contraception as detailed in Appendix 2 of 
this protocol during the treatment period and for at least 14 weeks after the 
last dose of investigational product. Patients must also agree to refrain from 
donating sperm during this period.  
b. Female patients:   
A female patient is eligible to participate if she is not pregnant (see Appendix 
2), not breastfeeding, and at least 1 of the following conditions applies:  
i. 
Not a woman of childbearing potential (WOCBP) as defined in Appendix 
2; OR 
ii. 
A WOCBP who agrees to follow the contraceptive guidance in Appendix 
2 during the treatment period and for at least 5 weeks after the last dose 
of investigational product. 
Informed Consent  
13. Capable of giving signed informed consent, which includes compliance with the 
requirements and restrictions listed in the Informed Consent Form (ICF) and in this 
protocol.  
14. Patient is willing to be compliant with study procedures, including completing the 
daily electronic diary during the Run-in Period and throughout the study. 
5.2 
Exclusion Criteria 
An individual for whom any of the following criteria apply will be excluded from 
participation in this study: 
Gastrointestinal-related Medical Conditions  


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1. At the time of screening, patient has a diagnosis of an IBS subtype other than IBS-D,
based on Rome IV criteria (
). Based on stool patterns on days that 
the patient experiences symptoms, other IBS subtypes are defined as follows:   
a. IBS-C: hard or lumpy stools ≥ 25% of bowel movements and loose or watery 
stools ≤ 25% of bowel movements;
b. IBS-M: hard or lumpy stools ≥ 25% of bowel movements and loose or watery 
stools ≥ 25% of bowel movements;
c. IBS-U: hard or lumpy stools ≤ 25% of bowel movements and loose or watery 
stools ≤ 25% of bowel movements.
2. Patient has a history of inflammatory or immune-mediated GI disorders including 
(but not limited to) inflammatory bowel disease (i.e., Crohn's disease, ulcerative 
colitis, microscopic colitis, and celiac disease).
3. Patient has had an episode of diverticulitis within 3 months prior to Screening.
4. Patient has a history of intestinal obstruction, stricture, toxic megacolon, GI 
perforation, fecal impaction, gastric banding, bariatric surgery, adhesions, ischemic 
colitis, or impaired intestinal circulation (e.g., aortoiliac occlusive disease).
5. Patient has any of the following surgical history:
a. Cholecystectomy with ANY history of post-cholecystectomy biliary tract pain. 
A successful cholecystectomy with no postoperative biliary tract pain is not 
exclusionary;
b. Any abdominal surgery within the 3 months prior to Screening; 
c. Major gastric, esophageal, hepatic, pancreatic, or intestinal surgery 
(appendectomy, hemorrhoidectomy, or polypectomy greater than 3 months post-
surgery are allowed).
6. Patient has a history or current evidence of laxative abuse. 
Other Medical Conditions
7. Patient has a history of a cardiovascular event, including stroke, myocardial 
infarction, congestive heart failure, or transient ischemic attack within 6 months prior 
to Screening.
8. Patient has a history of malignancy within 5 years prior to Screening (except 
squamous and basal cell carcinomas of the skin and cervical carcinoma in situ).
9. Patient has a history of alcohol abuse or binge drinking.
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10. Patient has uncontrolled hypertension, defined as systolic blood pressure 
> 180 mmHg or a diastolic blood pressure > 100 mmHg at the time of Screening. 
11. Patient has a history of significant hypersensitivity, intolerance, or allergy to any drug 
compound, food, or other substance, unless approved by the Investigator. 
Laboratory Assessments 
12. Fecal calprotectin ≥ 50μg/g. 
13. Hemoglobin A1c level ≥ 8.0% or a confirmed fasting plasma glucose level 
≥ 180 mg/dL. 
14. Confirmed alanine aminotransferase (ALT) > 2x upper limit of normal (ULN). 
15. Confirmed total bilirubin > ULN, unless the patient has a documented history of 
Gilbert’s syndrome. 
16. Confirmed QT interval corrected by Fridericia’s formula (QTcF) or QT interval 
corrected by Bazett’s formula (QTcB) > 500 msec. 
17. Evidence of active hepatitis B virus (HBV) infection, based on a positive hepatitis B 
surface antigen (HBsAg) screen. 
18. Evidence of hepatitis C virus (HCV) infection based on a positive HCV antibody 
screen (patients who have been successfully treated for HCV are eligible if 
an undetectable HCV viral load at least 6 months after completion of treatment can 
be demonstrated). 
19.  Human immunodeficiency virus (HIV)-1 or HIV-2 antibody positive. 
Prior/Concomitant Therapy  
20. Within 14 days of randomization, patient has used either of the following: 
x 5-hydroxytriptamine (5-HT)3 or 5-HT4 receptor antagonists (e.g., alosetron); 
x Any of the strong inhibitors of P-glycoprotein listed in Appendix 4. 
21. Within 14 days of randomization, patient has used any of the following:  
x Loperamide; (Note: loperamide can be used during the study as rescue 
medication based on protocol specified guidelines [see Section 6.7.1]);  
x Aspirin or aspirin-containing medications (> 325 mg of aspirin per day) or 
nonsteroidal anti-inflammatory drugs, when taken specifically for the 
symptoms of IBS; 
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x Cannabis-containing products; 
x Docusate; 
x Enemas; 
x GI preparations (including antacids containing aluminum or magnesium, 
antidiarrheal agents, antispasmodic agents, bismuth, peppermint oil, 
IBgard, FDgard, or prokinetic agents). 
22. Within 14 days of randomization, receipt of any prescribed or over-the-counter, 
systemic, herbal (including St. John’s wort), or topical medication, or any expectation 
of requiring use of such medication while participating in the study that, in the 
opinion of the Investigator, would interfere with study procedures, compromise 
safety, or the scientific integrity of the data.  
23. Patient has used, or is expected to use, the following antibiotics: 
x Rifaximin within 90 days prior to randomization;  
x Other oral antibiotics within 28 days of randomization (with the exception 
of topical antibiotics or a 1-day course with an antibiotic). 
However, a patient will be allowed to remain in the study should unplanned use of 
antibiotics other than rifaximin occur after the patient has been randomly assigned to 
study drug. 
24. Within 3 months prior to randomization, patient has had significant changes to his or 
her antidepressant regimen (i.e., addition of a new agent, discontinuation of a prior 
agent, significant modifications to the dose of a current medication). Patients on 
chronic stable doses of antidepressants will be allowed to participate in the study. 
As-needed use of buspirone and benzodiazepines for anxiety is permitted during the 
study; 
25. Within 30 days prior to randomization (or 5 half-lives, if known), the patient has 
received an investigational drug, or is currently enrolled in an investigational study. 
Other Exclusions  
26. Patient is unable to swallow solid oral dosage forms whole with the aid of liquid 
(patients may not chew, divide, dissolve, or crush the study drug). 
27. Patient has an elective surgery planned or expects to need elective surgery at any 
time during the study. 
28. Patient is pregnant or breastfeeding. 


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29. Patient has any medical or psychological disorder or condition that, in the opinion of 
the Investigator, would compromise the wellbeing of the patient or the study or 
prevent the patient from meeting or performing study requirements. 
30. Patient has poor peripheral venous access. 
31. Patient is an employee of the Investigator or study center with direct involvement in 
the proposed study or other studies under the direction of that Investigator or study 
center, as well as family members of the employees or the Investigator. 
5.3 
Lifestyle Considerations 
During this study, patients will be asked to abstain from strenuous exercise for 48 hours 
before each blood collection for clinical laboratory tests.  
Only on days when postdose PK blood samples are to be taken (see footnote f in the 
Schedule of Activities; Section 1.3), patients will be asked to refrain from consumption 
of food and water for 1 hour prior to and for 2 hours after administration of 
investigational product. Food and water may be consumed ad libitum at all other times 
during the study. Investigational product should be taken with approximately 240 mL 
water.  
5.4 
Screen Failures 
Screen failures are defined as patients who consent to participate in the clinical trial but 
are not subsequently randomized. A minimal set of screen failure information is required 
to ensure transparent reporting of screen failure patients, to meet the Consolidated 
Standards of Reporting Trials (CONSORT) publishing requirements and to respond to 
queries from regulatory authorities. Minimal information includes demography, screen 
failure details, eligibility criteria, and any SAE. 
Individuals who do not meet the criteria for participation in this trial (screen failure) may 
be rescreened with Medical Monitor approval.  


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6 
INVESTIGATIONAL PRODUCT 
6.1 
Investigational Products  
6.1.1 
Investigational Product Description 
Investigational 
Product Name:  
BOS-589 
Placebo 
Dosage Formulation:  
Tablet  
Tablet 
Unit Dose 
Strengths/Dosage 
Levels:  
50 mg (2 x 25-mg tablets) and 
200 mg (2 x 100-mg tablets) 
Visually matching placebo tablets 
across both BOS-589 dose levels 
Route 
of Administration: 
Oral 
Oral 
Dosing Instructions: 
Swallow whole with a glass of 
water, do not chew, divide, 
dissolve, or crush 
Swallow whole with a glass of 
water, do not chew, divide, 
dissolve, or crush 
Packaging 
and Labeling:  
Investigational product will be 
provided in white, opaque, 
high-density polyethylene (HDPE) 
bottles with a child-resistant 
closure. Each bottle will be labeled 
as per country requirement 
Investigational product will be 
provided in white, opaque, HDPE 
bottles with a child-resistant 
closure. Each bottle will be labeled 
as per country requirement 
Manufacturer:  
WuXi STA, Shanghai, China 
WuXi STA, Shanghai, China 
6.2 
Preparation/Handling/Storage/Accountability 
6.2.1 
Acquisition and accountability 
The Investigator or designee must confirm appropriate temperature conditions have 
been maintained during transit for all investigational product received and any 
discrepancies are reported and resolved before use of the investigational product.  
The Investigator, institution, or the head of the medical institution (where applicable) is 
responsible for investigational product accountability, reconciliation, and record 
maintenance (i.e., receipt, reconciliation, and final disposition records).  
6.2.2 
Product Storage and Stability 
All investigational product must be stored in a secure, environmentally controlled, and 
monitored (manual or automated) area in accordance with the labeled storage 
conditions with access limited to the Investigator and authorized site staff. 
Further guidance and information for the final disposition of unused investigational 
products are provided in the Pharmacy Manual. 


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6.3 
Measures to Minimize Bias 
This is a randomized, double-blinded, PBO-controlled study. Patients should be 
randomized as close as possible to the time of the planned first dose of study treatment.  
Randomization will be conducted via an interactive web response system (IWRS) in a 
1:1:1 ratio for patients to receive BOS-589 200 mg, BOS-589 50 mg, or PBO. Patients 
will be assigned to the treatment arms using a permuted block randomization stratified 
by site (44 patients per group). The randomization schedule will be generated by the 
Study Biostatistician and will be transferred to the IWRS team for loading into the 
system. The randomization file will be held by the Study Biostatistician until the end of 
the trial and when the database has been locked. 
The sponsor, site staff, and patients will be blinded to the treatment assignment. 
BOS-589 and PBO will be similar in appearance, and provided in white, opaque, 
high-density polyethylene (HDPE) bottles, each with a child-resistant closure. Each 
bottle will be labeled as per country requirement.  
Only in a medical emergency will a patient’s treatment assignment be unblinded, and 
this process will be performed and documented in the IWRS. Every effort should be 
made to contact the Medical Monitor to discuss unblinding prior to breaking the blind.  
6.4 
Dosing and Administration 
Only patients enrolled in the study may receive investigational product and only 
authorized site staff may supply or administer investigational product. 
Eligible patients will be randomized (1:1:1) to receive for a total of 4 weeks BOS-589 at 
1 of 2 dose levels or PBO bid. Patients will be randomized to the following cohorts:   
x Cohort 1 (High Dose): 2 x 100 mg tablets for the 200 mg BOS-589 bid orally  
x Cohort 2 (Low Dose): 2 x 25 mg tablets for the 50 mg BOS-589 bid orally 
x Cohort 3 (PBO): 2 x visually matched PBO tablets bid orally 
For each dose level and at each dosing timepoint, the 2 tablets are to be swallowed with 
approximately 240 mL room temperature water and are to be swallowed whole (i.e., not 
divided, crushed, dissolved, or chewed). On days when postdose PK blood samples are 
to be taken (see footnote f in the Schedule of Activities, Section 1.3, patients will be 
asked to refrain from consumption of food and water for 1 hour prior to and for 2 hours 
after administration of investigational product. Food and water may be consumed ad 
libitum at all other times during the study. 
Patients will be instructed to take the investigational product approximately every 
12 hours at approximately the same time each day. If a patient misses a dose at a given 
timepoint, and the time is within 4 hours of the regularly scheduled dosing time, the 
patient should be instructed to take the investigational product. If it has been longer than 


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4 hours, the patient should skip the missed dose at that timepoint and resume the 
regularly scheduled dosing schedule at the next scheduled timepoint.  
In the event of vomiting following administration, BOS-589 should not be taken until the 
next scheduled dose.  
6.5 
Dose modification 
Study treatment should be interrupted if the patient reports any treatment-related severe 
AE. Therapy should only be restarted after approval by the Medical Monitor.  
If therapy is restarted, the same dose should be administered as that prior to 
occurrence of the AE. If the patient is still having tolerability issues and an AE reoccurs, 
the dose may be reduced from 2 tablets of study treatment bid to 1 tablet of study 
treatment bid, without breaking the study blind.   
If another AE occurs after the study treatment dose reduction, study treatment will be 
permanently discontinued and the subject followed until the end of the study. 
6.6 
Investigational Product Compliance 
All patients in this study will commence therapy on site on Study Day 1; oral 
investigational product will be administered under clinic staff supervision. After patients 
are discharged to continue therapy at home, compliance will be assessed on 
subsequent visits by returned tablet count. Administration of investigational product and 
any deviation(s) from the prescribed dosage regimen should be recorded in the case 
report form (CRF) and Drug Accountability Record. 
6.7 
Concomitant Therapy 
Refer to Appendix 4. 
 for more details on which concomitant medications are 
permitted and which are excluded while patients are on study.  
Any prior or concomitant therapy (including over the counter or prescription medicines, 
vitamins and/or supplements) taken 28 days prior to the first dose of investigational 
product through the End-of-Study Visit must be recorded, along with:  
x Reason for use; 
x Dates of administration including start and end dates; 
x Dosage information including dose and frequency. 
The Medical Monitor should be contacted if there are any questions regarding 
concomitant or prior therapy.  
For this protocol, a prescription medication is defined as a medication that can be 
prescribed only by a properly authorized/licensed clinician. Medications to be reported 


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in the CRF are concomitant prescription medications, over-the-counter medications, 
vaccines, vitamins, and supplements. 
6.7.1 
Rescue Medicine 
During the double-blind Treatment Phase of the study, patients will be allowed to take 
loperamide rescue medication for the acute treatment of uncontrolled diarrhea. 
Loperamide at a unit dose of 2 mg may be taken once approximately every 6 hours with 
the following guidelines: 
x No more than 4 unit doses over a continuous 24-hour time period (8 mg/day); 
x No more than 7 unit doses over a continuous 48-hour time period (14 mg over 
2 days); 
x No more than 11 unit doses over a continuous 7-day time period. 
The use of loperamide rescue medication should be recorded electronically.  
6.8 
Intervention After the End of the Study  
BOS-589 will not be provided to study patients after the end of the study. 


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7 
DISCONTINUATION / WITHDRAWAL 
7.1 
Discontinuation of Investigational Product 
A patient may discontinue from study treatment at any time at his or her own request or 
may be withdrawn at any time at the discretion of the Investigator for safety, behavioral, 
or administrative reasons. Discontinuation from the investigational product does not 
mean discontinuation from the study, and remaining study procedures should be 
completed as indicated by the study protocol. If a clinically significant finding is identified 
(including, but not limited to, changes from baseline) after enrollment, the Investigator or 
qualified designee will determine if any change in patient management is needed. Any 
new clinically relevant finding will be reported as an AE. 
Upon investigational product discontinuation, patients should complete all procedures 
collected in the Day 29/End-of-Treatment visit as applicable per the Schedule of 
Activities (Section 1.3). 
7.2 
Discontinuation/Withdrawal from the Study 
A patient may withdraw from the study at any time at his or her own request or may be 
withdrawn at any time at the discretion of the Investigator for the following reasons: 
x If any clinical AE, laboratory abnormality, or other medical condition or situation 
occurs such that continued participation in the study would not be in the best 
interest of the patient; 
x Disease progression that requires discontinuation of the investigational product; 
x Pregnancy; 
x If the patient meets an exclusion criterion (either newly developed or not 
previously recognized) that precludes further study participation after consultation 
with the Medical Monitor; 
x Significant investigational product noncompliance; 
x Termination of the study by the sponsor. 
If the patient withdraws consent for disclosure of future information, the sponsor may 
retain and continue to use any data collected before such a withdrawal of consent.  
If a patient withdraws from the study, he or she may request destruction of any samples 
taken and not tested, and the Investigator must document this in the site study records.  
See Schedule of Activities, Section 1.3, for data to be collected at the time of study 
discontinuation and follow-up and for any further evaluations that need to be completed.  


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The reason for patient discontinuation or withdrawal from the study will be recorded on 
the CRF.  
Patients who sign the ICF and are randomized but do not receive the investigational 
product may be replaced. Once randomized, patients who are withdrawn from therapy 
before completing 28 days of dosing or are discontinued from the study for any reason 
will not be replaced.  
Clinical and/or laboratory criteria that may warrant study drug discontinuation.  
If a patient experiences any of the following clinical or laboratory events, study 
treatment should be immediately suspended: 
x Any SAE considered study drug-related;  
x No bowel movement for 7 days; 
x Any clinical evidence suggestive severe constipation, obstipation or intestinal 
obstruction; gastrointestinal hemorrhage, significant gastrointestinal infection; 
x Abnormal liver functions concerning for drug-induced as follows1:  
o Confirmed ALT > 5x ULN 
o Confirmed ALT > 3x ULN and any of the following signs or symptoms 
(fatigue, nausea, vomiting, right upper quadrant pain, fever, rash or 
eosinophilia)2 
o Confirmed ALT > 3x ULN and total bilirubin > 2x ULN or INR > 1.5 (should 
be repeated in 48-72 hours) 
1Should any of these events occur, a full evaluation of other causes of liver enzyme 
elevation should be undertaken and patients would be followed at regular intervals until 
labs return to normal/stabilize, or an alternate diagnosis is confirmed.   
2If patients develop asymptomatic elevations of the ALT of > 3x ULN and, if there are 
persistent elevations upon repeat testing, close observation should be implemented, 
and discontinuation of the drug should be considered. 
Depending on the nature, severity, and outcome, the Sponsor will decide to (and 
document with supporting rationale) either: 1) discontinue study drug; 2) request 
additional safety data; or 3) continue dosing at current dose or reduced dose (see 
Section 6.5). 
If 2 or more patients experience any of the above criteria (note: study drug-related SAEs 
must be in the same system organ class) the study enrollment will be halted.  The 
Sponsor, an independent GI physician not involved with the study, and the lead 
investigator will review available data to determine whether individual patient unblinding 


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is warranted, amendments are required for the protocol, and whether patients currently 
on treatment should also discontinue study drug.  A decision to continue (with or without 
modifications) or discontinue the study will be made.  All supporting rationale will be 
documented. 
7.3 
Lost to Follow-up 
A patient will be considered lost to follow-up if he or she repeatedly fails to return for 
scheduled visits and is unable to be contacted by the study site staff.  
The following actions must be taken if a patient fails to return to the clinic for a required 
study visit: 
x The site will attempt to contact the patient and reschedule the missed visit as soon 
as possible; counsel the patient on the importance of maintaining the assigned visit 
schedule; and ascertain if the patient wishes to and/or should continue in the study. 
x Before a patient is deemed lost to follow-up, the Investigator or designee will make 
every effort to regain contact with the patient (where possible, 3 telephone calls and, 
if necessary, a certified letter to the patient’s last known mailing address or local 
equivalent methods. These contact attempts should be documented in the patient’s 
medical record or study file.  
x Should the patient continue to be unreachable, he or she will be considered to have 
withdrawn from the study with a primary reason of lost to follow-up. 
 


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8 
STUDY ASSESSMENTS AND PROCEDURES 
Planned timepoints for all assessments are provided in the Schedule of Activities 
(Section 1.3). 
8.1 
Study Periods 
The study will comprise 3 phases:  
1. Pre-Treatment Phase (up to 4 weeks), in which all patients will be assessed for 
eligibility. The pre-treatment phase will consist of initial screening assessments 
(Visit 1) and a Run-in period (Visit 2 through Visit 3). 
2. A Treatment Phase (4 weeks), in which all patients will return to the clinical for 
randomization and start of treatment (Visit 3 – Day 1) followed by weekly visits 
(Visit 4 – Day 8 [± 1 day], Visit 5 – Day 15 [± 1 day], Visit 6 – Day 22 [± 1 day]) until 
Visit 7 – Day 29 (± 1 day), and  
3. A Follow-up Phase (2 weeks), in which patients who complete Visit 7, will be asked 
to return for an End-of-Study Follow-up visit (Visit 8 – Day 43 [± 3 days]) for 
assessment of treatment outcome (e.g., safety, durability of effect). 
8.2 
Screening Assessments 
After signing the ICF(s), the patient will undergo the initial screening assessments and 
procedures as described in the Schedule of Activities (Section 1.3) to determine 
eligibility.  
Eligible patients will enter the Run-in period. At the beginning of the Run-in period, 
patients will receive instructions for completing an electronic diary to collect daily 
information related to their IBS-D symptoms, their bowel function, and rescue 
medication use.  
Patients who are: 
x Compliant in completing the screening diary on a daily basis on at least 6 of the 
7 days during the week prior to randomization AND on at least 11 of the 14 days 
during the 2 weeks prior to randomization, and 
x Have an average WAP score in the past 24 hours of between 4.0 and 8.0 on a 
0 to 10 scale over the week prior to randomization, and  
x Have an average daily BSFS of ≥ 5.0 and at least 5 days with a BSFS score 
≥ 5.0 on a 1 to 7 scale over the week prior to randomization, and  
x Have an average daily IBS-GS score of ≥ 2.0 on a 0 to 4 scale over the week 
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x
Who have not used any loperamide rescue medication in the 2 weeks prior to 
randomization 
will be eligible for participation and immediate randomization into the double-blind 
treatment phase (i.e., all 5 diary conditions must be met to qualify for randomization). 
Eligible patients will be instructed to return to the study site for the Baseline visit on 
Day 1. 
If after 2 weeks, patients do not meet all of the 5 diary conditions required for entry, the 
run-in may be extended by up to one week in accordance with investigator judgement
(for a total run-in period of up to 3 weeks). 
8.3
Efficacy Assessments 
The efficacy objectives of the study are to evaluate the effect of BOS-589, relative to 
PBO after 4 weeks of therapy, on pain, defecation, and key IBS-D related signs and 
symptoms.
During the 4 weeks of the double-blind treatment phase, patients will be required to 
access their electronic diary every day, preferably at the same time each day, to record 
IBS-D symptom data and information related to their bowel function and rescue 
medication use.
8.3.1
Worst Abdominal Pain
Throughout the 4 weeks of the double-blind treatment phase, patients will be asked to 
rate their WAP in the past 24 hours. The patient-reported WAP in the past 24 hours will 
be recorded on a 0 to 10 scale, where 0 corresponds to no pain and 10 corresponds to 
worst imaginable pain. 
8.3.2
Bristol Stool Form Scale
Patients will be asked to record daily stool consistency according to the BSFS most 
representative of the past 24 hours. The patient-reported BSFS consistency score is 
based on a 1 to 7 scale where 1 corresponds to a hard stool and 7 corresponds to 
watery diarrhea (
). Please refer to Appendix 3.
8.3.3
IBS-D Global Symptom Score
Patients will be asked to record daily their overall IBS-D global symptoms in the prior
24 hours. The patient-reported daily IBS-GS is based on a 0 to 4 scale where: 
x
0 corresponds to no symptoms; 
x
1 corresponds to mild symptoms; 
x
2 corresponds to moderate symptoms; 
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x 3 corresponds to severe symptoms; and  
x 4 corresponds to very severe symptoms.  
8.3.4 
IBS Severity Score System 
Patients will be asked to complete 5 questions regarding the severity of their IBS. Each 
of the 5 questions generate a maximum score of 100, leading to a total possible score 
of 500.   
8.4 
Safety and Other Assessments 
8.4.1 
Adverse Events and Serious Adverse Events 
See Section 8.5. 
8.4.2 
Physical Examinations  
A complete physical examination (PE) will be performed at Screening. A symptom-
directed PE will be performed at the timepoints specified in the Schedule of Activities 
(Section 1.3) and as clinically indicated, and results recorded in the source document. 
Any clinically significant PE finding noted at Screening will be recorded as medical 
history. Any clinically significant PE finding noted at after enrollment will be reviewed for 
reporting as an AE (see Section 8.5). 
A complete PE will include, at a minimum, assessment of the cardiovascular, 
respiratory, GI, and neurological systems. A targeted PE will include, at a minimum, 
assessments of the skin, lungs, cardiovascular system, and abdomen (liver and spleen). 
Height and weight will also be measured and BMI will be calculated and the data 
recorded at the timepoints specified in the Schedule of Activities (Section 1.3). 
Investigators should pay special attention to clinical signs related to previous serious 
illnesses. 
8.4.3 
Vital Signs  
At the timepoints specified in the Schedule of Activities (Section 1.3) and before any 
blood sample collection, vital signs will be measured in a supine position after 5 minutes 
rest and will include oral temperature, systolic and diastolic blood pressure, pulse, and 
respiratory rate. Any clinically significant abnormal vital sign value will be recorded as an 
AE (see Section 8.5). 
8.4.4 
Electrocardiograms  
Single 12-lead ECGs will be performed at the timepoints specified in the Schedule of 
Activities (Section 1.3) and before any blood sample collection. Electrocardiogram 
interval measurements and interpretation (normal, abnormal not clinically significant, 


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abnormal clinically significant) will be recorded in the source document. Any clinically 
significant change in ECG interpretation will be recorded as an AE (see Section 8.5). 
8.4.5 
Clinical Safety Laboratory Assessments  
See Appendix 1 for the list of clinical laboratory tests to be performed and to the 
Schedule of Activities (Section 1.3) for the timing and frequency.   
The Investigator must review the laboratory report, document this review, and record in 
the AE section of the CRF any clinically relevant changes occurring during the study. 
The laboratory reports must be filed with the source documents. Clinically significant 
abnormal laboratory findings are those that are not associated with the underlying 
disease, unless judged by the Investigator to be more severe than expected for the 
patient’s condition.  
All laboratory tests with values considered clinically significantly abnormal during 
participation in the study should be repeated until the values return to normal or 
baseline or are no longer considered clinically significant by the Investigator or Medical 
Monitor. If such values do not return to normal/baseline within a period of time judged 
reasonable by the Investigator, the etiology should be identified and the sponsor 
notified.  
All protocol-required laboratory assessments, as defined in Appendix 1, must be 
conducted in accordance with the laboratory manual and the Schedule of Activities.   
If laboratory values from non-protocol-specified laboratory assessments performed at 
the institution’s local laboratory require a change in patient management or 
are considered clinically significant by the Investigator (e.g., SAE, or AE, or dose 
modification), then the results must be recorded in the CRF. 
8.5 
Adverse Events and Serious Adverse Events 
Adverse events will be reported by the patient (or, when appropriate, by a caregiver, 
surrogate, or the patient's legally authorized representative). 
The Investigator and any qualified designees are responsible for detecting and 
documenting events that meet the definition of an AE or SAE and are responsible for 
following all AEs. 
8.5.1 
Definition of Adverse Events 
An AE is any untoward medical occurrence in a patient or clinical study participant, 
temporally associated with the use of investigational product, whether or not considered 
related to the investigational product. An AE can therefore be any unfavorable and 
unintended sign (including an abnormal laboratory finding), symptom, or disease (new 
or exacerbated) temporally associated with the use of investigational product.   


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8.5.2 
Events of Special Interest  
Not applicable. 
8.5.3 
Definition of Serious Adverse Events 
If an event is not an AE per definition above, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, 
death because of progression of disease).  
A SAE is defined as any untoward medical occurrence that, at any dose:  
x Results in death; 
x Is life threatening;  
The term 'life threatening' in the definition of 'serious' refers to an event in which 
the patient was at risk of death at the time of the event. It does not refer to an 
event, which hypothetically might have caused death, if it were more severe.  
x Requires inpatient hospitalization or prolongation of existing hospitalization;   
In general, hospitalization signifies that the subject or patient has been detained 
(usually involving at least an overnight stay) at the hospital or emergency ward 
for observation and/or treatment that would not have been appropriate in the 
physician’s office or outpatient setting. Complications that occur during 
hospitalization are AEs. If a complication prolongs hospitalization or fulfills any 
other serious criteria, the event is serious. When in doubt as to whether 
“hospitalization” occurred or was necessary, the AE should be considered 
serious.   
Hospitalization for elective treatment of a pre-existing condition that did not 
worsen from baseline is not considered an AE;  
x Results in persistent disability/incapacity;  
The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions. This definition is not intended to include experiences of 
relatively minor medical significance such as uncomplicated headache, nausea, 
vomiting, diarrhea, influenza, and accidental trauma (e.g., sprained ankle) that 
may interfere with or prevent everyday life functions but do not constitute a 
substantial disruption.  
x Is a congenital anomaly/birth defect.  
Medical or scientific judgment should be exercised when deciding if SAE reporting is 
appropriate in other situations such as important medical events that may not be 
immediately life threatening or result in death or hospitalization, but may jeopardize the 


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patient or may require medical or surgical intervention to prevent any of the other 
outcomes listed in the above definition. These events should usually be considered 
serious. Examples of such events include invasive or malignant cancers, intensive 
treatment in an emergency room or at home for allergic bronchospasm, blood 
dyscrasias or convulsions that do not result in hospitalization, or development of drug 
dependency or drug abuse.  
8.5.4 
Classification of an Adverse Event 
8.5.4.1 
Assessment of Severity 
The Investigator will make an assessment of severity for each AE and SAE reported 
during the study and assign it to 1 of the following categories:   
x Mild: An event that is easily tolerated by the patient, causing minimal discomfort 
and not interfering with everyday activities;  
x Moderate: An event that causes sufficient discomfort and interferes with normal 
everyday activities;  
x Severe: An event that prevents normal everyday activities. An AE that is 
assessed as severe should not be confused with a SAE. Severe is a category 
utilized for rating the intensity of an event; and both AEs and SAEs can be 
assessed as severe.  
An event is defined as ‘serious’ when it meets at least 1 of the predefined outcomes as 
described in the definition of an SAE, NOT when it is rated as severe.  
8.5.4.2 
Assessment of Causality 
All AEs must have their relationship to the investigational product and/or study 
participation assessed by the Investigator who examines and evaluates the patient 
based on temporal relationship and his or her clinical judgment. Alternative causes, 
such as underlying disease, concomitant therapy, and other risk factors, as well as the 
temporal relationship of the event to investigational product administration will be 
considered and investigated. The Investigator will also consult the IB in his or her 
assessment. 
The degree of certainty about causality will be graded using the categories below:  
x Related – The AE is known to occur with the investigational product, there is a 
reasonable possibility that the investigational product caused the AE, or there is a 
temporal relationship between the investigational product and event. Reasonable 
possibility means that there is evidence to suggest a causal relationship between 
the investigational product and the AE. 
x Not Related – There is not a reasonable possibility that the administration of the 
investigational product caused the event, there is no temporal relationship 


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between the investigational product and event onset, or an alternate etiology has 
been established. 
For each AE, the Investigator must document in the medical notes that he or she has 
reviewed the event and has provided an assessment of causality. The Investigator may 
change his or her opinion of causality in light of follow-up information and updated 
causality assessment reported. 
8.5.4.3 
Expectedness  
The Investigator will be responsible for determining whether an AE is expected or 
unexpected. An AE will be considered unexpected if the nature, severity, or frequency of 
the event is not consistent with the risk information previously described for the 
investigational product. 
8.5.5 
Time Period and Frequency for Event Assessment and Follow-up 
All AEs will be collected from the signing of the ICF until the End-of-Study Follow-up 
visit.  
Whenever possible, all AEs should be followed until satisfactory resolution or until the 
site Investigator deems the event to be chronic or the patient is stable.   
All SAEs will be recorded and reported to the sponsor or designee within 24 hours, 
whether considered investigational product related or not, and must include an 
assessment of if there is a reasonable possibility that the investigational product caused 
the event. The Investigator will also submit any updated SAE data to the sponsor within 
24 hours of it being available.  
Investigators are not obligated to actively seek AEs or SAEs after conclusion of the 
study participation. However, if the Investigator learns of any SAE, including a death, at 
any time after a patient has been discharged from the study, and he or she considers 
the event to be reasonably related to the investigational product or study participation, 
the Investigator must promptly notify the sponsor. 
8.5.6 
Adverse Event Reporting  
At each study visit, patients will be evaluated for new AEs and the status of existing 
AEs. Care will be taken not to introduce bias when evaluating for AEs. The Investigator 
should use open-ended questions when soliciting information from a patient regarding 
AEs, followed by appropriate questions that clarify the patient’s verbatim description of 
AEs or change in concomitant medications.  
When an AE occurs, it is the responsibility of the Investigator to review all 
documentation (e.g., hospital progress notes, laboratory reports, and diagnostics 
reports) related to the event. The Investigator will attempt to establish a diagnosis of the 
event based on signs, symptoms, and/or other clinical information. Whenever possible, 
the diagnosis (not the individual signs/symptoms) will be documented as the AE. 


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The Investigator will then record all relevant AE information in the CRF. It is not 
acceptable for the Investigator to send photocopies of the patient’s medical records in 
lieu of completion of the CRF page. New or updated information will be recorded in the 
originally completed CRF. However, there may be instances when copies of medical 
records for certain cases are requested. In this case, all patient identifiers, with the 
exception of the patient number, will be redacted on the copies of the medical records 
before submission. 
If a patient dies during participation in the study or during a recognized follow-up period, 
the Investigator will provide a copy of any postmortem findings, including histopathology 
when available. 
If a site receives report of a new SAE from a study patient or receives updated data on 
a previously reported SAE after database lock of the electronic data capture (EDC) 
system, then the site can report this information on a paper SAE form or to the sponsor 
by telephone. See the Study Manual for additional information on SAE reporting. 
The study sponsor, or designee, will be responsible for notifying all applicable regulatory 
authorities of any required safety events in compliance with country-specific regulatory 
requirements. In addition, the sponsor must notify applicable regulatory authorities and 
all participating Investigators of suspected unexpected serious adverse reactions 
(SUSARs), from clinical trials or any other source. 
An Investigator who receives an Investigator safety report describing a SAE or other 
specific safety information (e.g., summary or listing of SAEs) from the sponsor will 
review and then file it along with the IB and will notify the Institutional Review Board/ 
Independent Ethics Committee (IRB/IEC), if appropriate according to local 
requirements. 
8.5.7 
Reporting of Pregnancy  
Pregnancy itself is not regarded as an AE unless there is a suspicion that the 
investigational product may have interfered with the effectiveness of a contraceptive 
medication. Pregnancy in a patient’s partner is not considered an AE. Abnormal 
pregnancy outcomes (e.g., spontaneous abortion, fetal death, stillbirth, congenital 
anomalies, ectopic pregnancy) are considered SAEs. Elective abortions without 
complications should not be handled as AEs; however, an induced therapeutic abortion 
to terminate a pregnancy because of complications or medical reasons must be 
reported as an SAE. The underlying medical diagnosis for this procedure should be 
reported as the SAE term. A spontaneous abortion in a study patient is always 
considered an SAE. 
All pregnancies in female patients and, if indicated, pregnancies in female partners of 
male patients, will be collected after the start of investigational product and until 5 
weeks after the last dose. Pregnancy outcomes, defined as the first well baby visit for 
live births, or spontaneous or induced abortion, will be documented even after the 
patient is withdrawn from or completes the study. 


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If a pregnancy is reported, the Investigator should inform the sponsor within 24 hours of 
learning of the pregnancy and should follow the procedures outlined in Appendix 2. 
8.6 
Treatment of Overdose 
There are no data on overdose because this is the first study of BOS-589 in patients 
and, in a prior study in healthy human volunteers, no evidence of overdose was 
reported. There is no definition of what constitutes an overdose and no known antidote. 
Any patient who receives a higher dose than that intended (i.e., more than 200 mg per 
dose; more than 400 mg in a given day) should be monitored closely, managed with 
appropriate supportive care, and followed up appropriately. If possible, a blood sample 
for PK should be collected as soon as is feasible from any patient who takes a higher 
dose than that intended. 
If AEs or SAEs are reported and are considered related to a patient receiving a higher 
dose than intended, dosing should be interrupted if deemed necessary by the 
Investigator and the case discussed further with the Medical Monitor. 
8.7 
Pharmacokinetics 
Plasma samples of approximately 4 mL will be collected for measurement of plasma 
concentrations of BOS-589 at the following times and as specified in the Schedule of 
Activities (Section 1.3).  
x On Day 1 at predose and at 0.5, 1, 2, and 4 hours postdose.  
x On Day 15 at predose and at 0.5, 1, 2, and 4 hours postdose. If samples cannot 
be obtained on Day 15, then samples should be obtained on Day 22.  
x A predose sample will be obtained on all other Treatment Study Visits. 
Allowable sampling windows for PK blood draws will be 30 minutes prior to dosing for 
the predose sample, then ± 5 minutes for the 0.5 and 1 hour postdose timepoints, and 
± 10 minutes for the 2 and 4 hours postdose timepoints. 
Instructions for the collection and handling of biological samples will be provided by the 
sponsor. The actual time of dosing and actual date and time (24-hour clock time) of 
each sample will be recorded. 
Drug concentration information that may unblind the study will not be reported to 
investigative sites or blinded personnel until the study has been unblinded. 
Any changes in the timing or addition of timepoints for any planned study assessments 
must be documented and approved by the relevant study team member and then 
archived in the sponsor and site study files but will not constitute a protocol amendment. 
The IRB/IEC will be informed of any safety issues that require alteration of the safety 
monitoring scheme or amendment of the ICF. 


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The following PK parameters will be estimated from the plasma concentrations of 
BOS-589 using either traditional noncompartmental methods or a population-PK model:
AUC0-t 
Area under the plasma concentration versus time curve from time zero to 
the last quantifiable sample;
AUC0-4
Area under the plasma concentration versus time curve from time zero to 
4 hours postdose;
Cmax
Maximum observed plasma concentration;
Cmin
Minimum observed plasma concentration.
In addition to parameters above, the AUC from time zero to 4 hours postdose (AUC0-12) 
may be calculated using predose concentrations other than Day 1 predose to impute a 
12-hour concentration. Details will be provided in the PK analysis plan.
8.8
Biomarkers/Pharmacodynamics
9
STATISTICAL CONSIDERATIONS
9.1
Timing of Analyses
Formal hypothesis testing will occur at the end of the study and post database lock. 
There are no planned Data, Safety, or Adjudication Committees for this study and no 
planned statistical interim analyses.
9.2
Sample Size Determination
This is a phase 2a study in which a hierarchical hypothesis test will be employed for the 
primary efficacy endpoint. The primary endpoint is defined as a change in the 24-hour 
WAP score at Day 29 compared to baseline (averaged over the week prior to each 
respective timepoint). Mean change from baseline will be measured as a continuous 
variable and compared between treatment groups in a hierarchical testing order using a 
t-test of equal variances and a two-tailed alpha of 0.05. 
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The sample size was based on the first hypothesis test in the hierarchical plan, with 
80% power, two-sided alpha of 0.05, and a standard deviation (SD) of 3 points on the 
NRS to detect a 1.6 minimum change between treatment groups (N = 120 patients). To 
account for attrition and a potential higher PBO rate, a total of 132 patients will be 
randomized to one of 3 treatment groups in a 1:1:1 ratio (44 patients per treatment 
group). 
9.3 
Populations for Analyses 
1. 
The Intention-to-Treat (ITT) analysis dataset will comprise all randomized 
patients;  
2. 
The Per-protocol Analysis Dataset will comprise patients in the ITT Analysis 
Dataset who received at least 1 dose of investigational product and did not have 
any major protocol deviations;  
3. 
The Safety Analysis Dataset will comprise all patients who received at least 
1 dose of investigational product; 
4. 
The Pharmacokinetic Analysis Dataset will comprise all randomized patients who 
received the active study drug and have available serum time concentration data. 
9.4 
Statistical Analyses 
Detailed methodology for descriptive and inferential statistical analyses of the data 
collected in the study will be documented in the Statistical Analysis Plan (SAP). The 
SAP will be prepared by the Study Biostatistician and agreed upon by the Sponsor. The 
SAP will be finalized and approved prior to database lock. The SAP will take 
precedence over the protocol for details regarding the statistical analyses to be 
conducted for the study. In addition to the SAP, other graphical representations of the 
results may be produced after review of the data (post hoc). Any major modifications of 
the Primary Endpoint’s definition and/or its analysis will be reflected in a protocol 
amendment. 
In general, descriptive statistical methods will be used to summarize the data from this 
study. Unless stated otherwise, the term “descriptive statistics” refers to number of 
patients (n), mean, median, SD, minimum, and maximum for continuous data, and 
frequencies and percentages for categorical data. 
All statistical analyses will be performed using Statistical Analysis System (SAS®) 
software Version 9.4 or higher.   
9.4.1 
Efficacy Analyses 
9.4.1.1 
Primary Endpoint 
This is a proof-of-principle study in which a hierarchical hypothesis test will be employed 
for the primary efficacy endpoint, defined as a change in the 24-hour WAP score at 


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Day 29 compared to baseline (averaged over the week prior to each respective 
timepoint). Mean change from baseline will be measured as a continuous variable and 
compared between treatment groups in the following testing order using a t-test of equal 
variances and a two-tailed alpha of 0.05: 
Hypothesis 1:  
Active Treatment Group (Cohort 1 + Cohort 2) versus PBO (Cohort 3); 
If statistically significant at P < 0.05, then proceed to Hypothesis 2. 
Hypothesis 2: 
 
Cohort 1 (High Dose) versus Cohort 3 (PBO); 
If statistically significant at P < 0.05, then proceed to Hypothesis 3. 
Hypothesis 3: 
 
Cohort 2 (Low Dose) versus Cohort 3 (PBO); 
If statistically significant at P < 0.05 then, proceed to Hypothesis 4. 
Hypothesis 4: 
 
Cohort 1 (High Dose) versus Cohort 2 (Low Dose). 
9.4.1.2 
Secondary Endpoint 
Mean daily BSFS score (stool type) and mean daily frequency of bowel movements will 
be tabulated and summarized by study day for each treatment for the ITT and 
Per-protocol Analysis Dataset populations. Time profiles and box plots will also be 
presented by study day and by treatment group. Pooled active data may be compared 
to PBO for the analyses, and also by cohort. 
9.4.1.3 
Other Efficacy Endpoints 
Scores from IBS-GS and IBS-SS questionnaires will be summarized for the ITT and 
Per-protocol Analysis Dataset populations. Pooled active data may be compared to 
PBO for the analyses, and also by cohort. 
9.4.1.4 
Missing Data 
Study implementation procedures and training will assist in limiting the amount of 
missing data in the trial. However, sensitivity analyses may be performed when 
indicated for the study endpoint and the amount of missing data to be handled. Details 
of the statistical methods for handling missing data will be described in the SAP.  


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9.4.2 
Safety Analyses 
9.4.2.1 
Adverse Events 
Adverse events will be recorded from the time of consent until the Day 43/End-of-Study 
Follow-up visit. The number and percentage of patients with AEs will be displayed by 
system organ class and preferred term using Medical Dictionary for Regulatory Activities 
(MedDRA) Version 20.0 or higher, by study treatment. Summaries in terms of severity 
and relationship to investigational product will also be provided. All SAEs will be 
summarized in a similar manner. Patient listings of all AEs causing discontinuation of 
investigational product and all SAEs will be produced. 
All AEs will be listed for individual patients showing both verbatim and preferred terms. 
Separate summaries of treatment-emergent SAEs and treatment-emergent AEs 
(TEAEs) related to investigational product will be generated. 
Any event reported on the CRF that occurs on at the time of or after the initiation of 
investigational product is defined as treatment-emergent. Additionally, an AE that is 
reported to have started on Day 1 without an associated onset time will be assumed to 
have occurred after the initiation of investigational product. Hence, AEs occurring on 
Day 1 with no associated onset time will be assumed to be treatment emergent. 
Serious AEs associated with a protocol-specified procedure and occurring after the time 
of consent but before administration of first dose of investigational product will be 
defined as non-treatment-emergent AEs (NTEAEs). 
All safety analyses will be performed in the Safety Analysis Dataset. 
9.4.2.2 
Clinical Laboratory Evaluations 
Laboratory data will be listed for all patients. Laboratory data will be reviewed by the 
Investigator as they become available. All abnormal laboratory results will be evaluated 
by the Investigator as either clinically significant or not clinically significant. 
Descriptive summaries of clinical laboratory results will be presented by date and time 
of collection. The number and percentage of patients experiencing treatment-emergent 
graded toxicities will be summarized by treatment arm and severity grade. Laboratory 
toxicity shifts from baseline to post-baseline assessments will be summarized by 
treatment arm. Changes from baseline in laboratory tests will be summarized for each 
treatment arm. 
9.4.2.3 
Twelve-lead Electrocardiograms, Vital Signs, and Physical 
Examinations 
Twelve-lead ECG and vital signs data will be listed and summarized.  
Physical examination findings will be captured and listed as part of a patient’s medical 
history or as AEs as applicable. 


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9.4.3 
Other Analyses 
9.4.3.1 
Pharmacokinetic Analysis 
A full description of the methods for evaluating the PK of BOS-589 in this patient 
population will be provided in a PK data analysis plan. The PK data from this study may 
be combined with data from other studies to complete a population-PK analysis. 
A full list of PK parameters to be calculated is included in Section 8.7. 
9.4.4 
Baseline Descriptive Statistics 
Demographic and baseline characteristics will be summarized for all patients overall 
and by treatment arm. Summary statistics (e.g., number of patients, mean, median, SD, 
and range) will be generated for continuous variables (e.g., age and weight) and the 
number and percentage of patients within each category will be presented for 
categorical variables (e.g., gender, ethnicity, race). 
A detailed description of patient disposition will be provided. It will include: 
x A summary of overall patient enrollment status (consented, screened, screen 
failures, replacements, randomized); 
x A summary of patients who discontinued the study; 
x An account of identified protocol deviations. 
All patients who are consented for the study will be accounted for in the summation. The 
number of patients who do not qualify for certain analysis populations will be 
summarized. 
9.4.5 
Subgroup Analyses 
No subgroup analyses are powered for the study. Subgroup analyses may be 
performed for descriptive purposes as described in the SAP. 
9.5 
Planned Interim Analyses  
No formal efficacy interim analysis is planned for the purposes of statistically comparing 
the data prior to the completion of the study. 


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10 
SUPPORTING DOCUMENTATION AND OPERATIONAL 
CONSIDERATIONS 
10.1 Regulatory, Ethical, and Study Oversight Considerations 
10.1.1 
Regulatory and ethical considerations 
This study will be conducted in accordance with the protocol and with the following: 
x Consensus ethical principles derived from international guidelines including the 
Declaration of Helsinki and Council for International Organizations of Medical 
Sciences International Ethical Guidelines; 
x Applicable International Conference on/Council for Harmonisation (ICH) Good 
Clinical Practice (GCP) Guidelines; 
x Applicable laws and regulations. 
The protocol, protocol amendments, ICF, IB, and other relevant documents 
(e.g., advertisements) must be submitted, and reviewed and approved as necessary, to 
the relevant Competent Authorities, IRBs and IECs before the study is initiated.  
Any amendments to the protocol will require regulatory approval before implementation 
of changes made to the study design, except for changes necessary to eliminate an 
immediate hazard to study patients.  
The Investigator will be responsible for the following: 
x Providing written summaries of the status of the study to the IRB/IEC annually or 
more frequently in accordance with the requirements, policies, and procedures 
established by the IRB/IEC; 
x Notifying the IRB/IEC of SAEs or other significant safety findings as required by 
IRB/IEC procedures; 
x 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/IEC, 
European regulation 536/2014 for clinical studies (if applicable), and all other 
applicable local regulations. 
10.1.2 
Informed Consent/Assent Process 
An initial sample ICF is provided for the Investigator to prepare the informed consent 
document to be used at his or her site. Updates to the sample ICF are to be 
communicated formally in writing from the Sponsor or Sponsor’s designee to the 
Investigator. The written ICF is to be prepared in the language of the potential patient 
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The Investigator or his/her representative will explain the nature of the study to the 
patient or his/her legally authorized representative and answer all questions regarding 
the study.  
Patients must be informed that their participation is voluntary. Patients or their legally 
authorized representative will be required to sign a statement of informed consent, 
and/or assent, that meets the requirements of 21 CFR 50, local regulations, ICH 
guidelines, Health Insurance Portability and Accountability Act requirements, where 
applicable, and the IRB/IEC or study center.  
The medical record must include a statement that written informed consent was 
obtained before the patient was enrolled in the study and the date the written consent 
was obtained. The authorized person obtaining the informed consent must also sign the 
ICF. 
Patients must be re-consented to the most current version of the ICF(s) during their 
participation in the study if one becomes available.  
A copy of the ICF(s) must be provided to the patient or the patient’s legally authorized 
representative.  
The Investigator is also responsible for asking the patient if the patient has a primary 
care physician and if the patient agrees to have his/her primary care physician informed 
of the patient’s participation in the clinical study if it is a local requirement. If the patient 
agrees to such notification, the Investigator is to inform the patient’s primary care 
physician of the patient’s participation in the clinical study. If the patient does not have a 
primary care physician and the Investigator will be acting in that capacity, the 
Investigator is to document such in the patient’s medical record. 
If a patient is unable to read or if a legally acceptable representative is unable to read, 
an impartial witness should be present during the entire informed consent discussion. 
After the written ICF and any other written information to be provided to patients, is read 
and explained to the patient or the patient’s legally acceptable representative, and after 
the patient or the patient’s legally acceptable representative has orally consented to the 
patient’s participation in the trial and, if capable of doing so, has signed and personally 
dated the ICF, the witness should sign and personally date the consent form. By signing 
the consent form, the witness attests that the information in the consent form and any 
other written information was accurately explained to, and apparently understood by, 
the patient or the patient’s legally acceptable representative, and that informed consent 
was freely given by the patient or the patient’s legally acceptable representative (Refer 
to ICH GCP guideline, Section 4.8.9). 
 
10.1.3 
Confidentiality and Privacy  
Patient confidentiality and privacy is strictly held in trust by the participating 
Investigators, their staff, and the sponsor and its designees. This confidentiality is 


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extended to cover testing of biological samples and genetic tests in addition to the 
clinical information relating to patients. Therefore, the study documentation, data, and 
all other information generated will be held in strict confidence. No information 
concerning the study or the study data will be released to any unauthorized third party 
without prior written approval of the sponsor.  
Study patient research data will not include the patient’s contact or identifying 
information. Rather, individual patients and their research data will be identified by a 
unique study identification number. Any patient records or datasets that are transferred 
to the sponsor will contain the identifier only; patient names or any information which 
would make the patient identifiable will not be transferred. Research data will be used in 
accordance with local data protection laws. 
The study monitor, other authorized representatives of the sponsor, representatives of 
the IRB/IEC or regulatory agencies may inspect all documents and records required to 
be maintained by the Investigator, including but not limited to, medical records (office, 
clinic, or hospital) and pharmacy records for the patients in this study. The clinical study 
site will permit access to such records. 
10.1.4 
Quality Assurance and Quality Control 
Quality control (QC) procedures will be implemented beginning with the data entry 
system and data QC checks that will be run on the database will be generated. Any 
missing data or data anomalies will be communicated to the site(s) for 
clarification/resolution. 
Following written Standard Operating Procedures, the monitors will verify that the 
clinical trial is conducted, data are generated, and biological specimens are collected, 
documented (recorded), and reported in compliance with the protocol, ICH GCP, and 
applicable regulatory requirements (e.g., Good Laboratory Practices, Good 
Manufacturing Practices).  
The investigational site will provide direct access to all trial related sites, source 
data/documents, and reports for the purpose of monitoring and auditing by the sponsor, 
and inspection by local and regulatory authorities. 
10.1.5 
Data Handling and Record Keeping  
10.1.5.1 
Data Collection and Management Responsibilities  
Data collection is the responsibility of the clinical trial staff at the site, under the 
supervision of the site Investigator. The Investigator is responsible for ensuring the 
accuracy, completeness, legibility, and timeliness of the data reported. 
The Investigator must maintain adequate source documentation. All source documents 
should be completed in a neat, legible manner to ensure accurate interpretation of data.   


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Clinical data (including AEs, concomitant medications, and expected adverse reactions 
data) will be entered into a 21 CFR Part 11-compliant EDC system or transmitted 
electronically (e.g., laboratory data). The data system(s) includes password protection 
and internal quality checks, such as automatic range checks, to identify data that 
appear inconsistent, incomplete, or inaccurate. Clinical data will be entered directly from 
the source documents. Data recorded in the CRF should be consistent with the data 
recorded on the source documents or the discrepancies must be explained.  
The sponsor or designee is responsible for the data management of this study, including 
quality checking of the data. The sponsor assumes accountability for actions delegated 
to other individuals (e.g., Contract Research Organizations). 
Study monitors will perform ongoing source data verification to confirm that data entered 
into the CRF by authorized site personnel are accurate, complete, and verifiable from 
source documents; that the safety and rights of patients 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.  
10.1.5.2 
Study Records Retention  
Records and documents, including signed ICFs, pertaining to the conduct of this study 
must be retained for a minimum of 2 years after the last approval of a marketing 
application in an ICH region and until there are no pending or contemplated marketing 
applications in an ICH region or until at least 2 years have elapsed since the formal 
discontinuation of clinical development of the investigational product. These documents 
should be retained for a longer period, however, if required by local regulations.  
No records will be destroyed without the written consent of the sponsor. No records may 
be transferred to another location or party without written notification to the sponsor.   
It is the responsibility of the sponsor to inform the Investigator when these documents 
no longer need to be retained. 
10.1.6 
Study and Site Discontinuation and Closure 
The sponsor or its designee reserves the right to suspend or prematurely terminate a 
study site or study at any time for any reason and at the sole discretion of the sponsor.  
An Investigator may initiate site closure at any time, provided there is reasonable cause 
and sufficient notice is given to the sponsor in advance of the intended termination.   
Written notification, documenting the reason for site or study suspension or termination, 
will be provided by the suspending or terminating party to study participants, 
Investigator, funding agency, the Investigational New Drug sponsor, and regulatory 
authorities.   
If the study is prematurely terminated or suspended, the Investigator will promptly 
inform study participants, the IRB/IEC, and sponsor, and will provide the reason(s) for 


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the termination or suspension. Study participants will be contacted, as applicable, and 
be informed of changes to study visit schedule.  
Circumstances that may warrant termination or suspension include, but are not limited 
to: 
x Determination of unexpected, significant, or unacceptable risk to participants; 
x Demonstration of efficacy that would warrant stopping; 
x Insufficient compliance to the protocol, regulatory requirements, or GCP 
guidelines; 
x Data that are not sufficiently complete and/or evaluable; 
x Determination that the primary endpoint has been met; 
x Determination of futility; 
x Inadequate recruitment of patients. 
10.1.7 
Dissemination of Clinical Study Data  
A Clinical Study Report (CSR) will be prepared in accordance with the ICH guideline on 
structure and contents of CSRs and any applicable regulatory and legal requirements. 
10.1.8 
Publication Policy 
The results of this study may be published or presented at scientific meetings. If this is 
foreseen, the Investigator agrees to submit all manuscripts or abstracts to the sponsor 
before submission. This allows the sponsor to protect proprietary information and to 
provide comments.  
The sponsor will comply with the requirements for publication of study results. In 
accordance with standard editorial and ethical practice, the sponsor will generally 
support publication of multicenter studies only in their entirety and not as individual site 
data. In this case, a coordinating Investigator will be designated by mutual agreement. 
Authorship will be determined by mutual agreement and in line with International 
Committee of Medical Journal Editors authorship requirements. 


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11
REFERENCES 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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U.S. Department of Health and Human Services, Food and Drug Administration, Center 
for Drug Evaluation and Research (May 2012). Guidance for Industry, Irritable Bowel 
Syndrome – Clinical Evaluation of Drugs for Treatment. Retrieved from 
https://www.fda.gov/downloads/Drugs/Guidances/UCM205269.pdf.
 
 
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12 
APPENDICES


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Appendix 1. Clinical Laboratory Tests
The tests detailed in Table 2 will be performed by the central laboratory with the 
exception of erythrocyte sedimentation rate and urine pregnancy tests.  The central 
laboratory will provide kits and instructions to perform these assessments on site.
Local laboratory results are only required in the event that the central laboratory results 
are not available in time for either investigational product administration and/or 
response evaluation. If a local sample is required, it is important that the sample for 
central analysis is obtained at the same time. Additionally, if the local laboratory results 
are used to make either an investigational product decision or response evaluation, the 
results must be entered into the CRF.
Protocol-specific requirements for inclusion or exclusion of patients are detailed in 
Section 5 of the protocol.
Additional tests may be performed at any time during the study as determined 
necessary by the Investigator or required by local regulations.
Pregnancy testing: Refer to Section 5.1 for Screening pregnancy criteria.
Table 2.
Protocol-required Safety Laboratory Assessments
Laboratory 
Assessments
Parameters
Hematology
Platelet count
RBC Indices:
Mean corpuscular 
volume
Mean corpuscular 
Hemoglobin
% Reticulocytes
White blood cell count 
with differential:
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils
Red blood cell (RBC) count
Hemoglobin
Hematocrit
Clinical 
Chemistry
Blood 
urea
nitrogen
Potassium
Aspartate 
aminotransferase/Serum 
glutamic oxaloacetic 
transaminase
Total and direct 
bilirubin
Creatinine
Sodium
Alanine 
aminotransferase/Serum 
glutamic pyruvic 
transaminase
Total protein
Glucose 
(fasted)
Calcium
Alkaline phosphatase
Routine 
Urinalysis
Specific gravity
pH, glucose, protein, blood, ketones, bilirubin, urobilinogen, nitrite, leukocyte esterase 
by dipstick
Microscopic examination (if blood or protein is abnormal)
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Table 2. 
Protocol-required Safety Laboratory Assessments 
Laboratory 
Assessments 
Parameters 
Other 
Screening 
Tests 
Hemoglobin A1c 
Follicle-stimulating hormone and estradiol (as needed in women of nonchildbearing 
potential only) 
Urine human chorionic gonadotropin (hCG) pregnancy test (as needed for women of 
childbearing potential) 
Serology (HIV antibody, hepatitis B surface antigen, and hepatitis C virus antibody)  
serum tissue transglutaminase IgA antibody (tTG-IgA) and IgA (if applicable)   
 
Investigators must document their review of each laboratory safety report. 
Laboratory results that could unblind the study will not be reported to investigative sites 
or other blinded personnel until the study has been unblinded. 


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Appendix 2. Contraceptive Guidance and Collection of Pregnancy Information 
Definitions: 
Woman of Childbearing Potential (WOCBP) 
A woman is considered fertile following menarche and until becoming postmenopausal 
unless permanently sterile (see below). 
Women in the following categories are not considered WOCBP 
1. Premenarchal 
2. Premenopausal female with 1 of the following: 
a. Documented hysterectomy 
b. Documented bilateral salpingectomy 
c. Documented bilateral oophorectomy 
Note: Documentation can come from the site personnel’s review of the patient’s medical 
records, medical examination, or medical history interview. 
3. Postmenopausal female 
a. A postmenopausal state is defined as no menses for 12 months without an 
alternative medical cause. A high follicle-stimulating hormone (FSH) level in the 
postmenopausal range may be used to confirm a postmenopausal state in 
women not using hormonal contraception or hormonal replacement therapy 
(HRT). However, in the absence of 12 months of amenorrhea, a single FSH 
measurement is insufficient.  
b. Females on HRT and whose menopausal status is in doubt will be required to 
use one of the nonestrogen hormonal highly effective contraception methods if 
they wish to continue their HRT during the study. Otherwise, they must 
discontinue HRT to allow confirmation of postmenopausal status before study 
enrollment. 
Contraception Guidance: 
Male patients. 
Male patients with female partners of childbearing potential are eligible to participate if 
they agree to ONE of the following during the treatment period and for at 
least 14 weeks after the last dose of investigational product:  
x Are abstinent from penile-vaginal intercourse as their usual and preferred lifestyle 
(abstinent on a long term and persistent basis) and agree to remain abstinent 


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x Agree to use a male condom plus partner use of a contraceptive method with a 
failure rate of < 1% per year, as described in Table 3, when having penile-vaginal 
intercourse with a WOCBP who is not currently pregnant 
In addition, male patients must refrain from donating sperm for the duration of the study 
and for 14 weeks after the last dose of investigational product. 
Male patients with a pregnant or breastfeeding partner must agree to remain abstinent 
from penile-vaginal intercourse or use a male condom during each episode of penile 
penetration for the duration of the study and for 14 weeks after the last dose of 
investigational product. 
Female patients. 
Female patients of childbearing potential are eligible to participate if they agree to use a 
highly effective method of contraception, as described in Table 3, consistently and 
correctly.


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Table 3. 
Highly Effective Contraceptive Methods 
Highly Effective Contraceptive Methods That Are User Dependenta  
Failure rate of <1% per year when used consistently and correctly.  
Combined (estrogen and progestogen containing) hormonal contraception associated with inhibition of 
ovulationb  
x 
Oral  
x 
Intravaginal  
x 
Transdermal  
Progestogen only hormonal contraception associated with inhibition of ovulation 
x 
Oral  
x 
Injectable  
Highly Effective Methods That Are User Independenta  
Implantable progestogen only hormonal contraception associated with inhibition of ovulationb 
x 
Intrauterine device (IUD) 
x 
Intrauterine hormone-releasing system (IUS) 
x 
Bilateral tubal occlusion 
Vasectomized Partner  
A vasectomized partner is a highly effective contraception method, provided that the partner is the sole 
male sexual partner of the WOCBP and the absence of sperm has been confirmed. If not, an additional 
highly effective method of contraception should be used.  
Sexual Abstinence  
Sexual abstinence is considered a highly effective method only if defined as refraining from heterosexual 
intercourse during the entire period of risk associated with the investigational product. The reliability of 
sexual abstinence needs to be evaluated in relation to the duration of the study and the preferred and 
usual lifestyle of the patient.  
NOTES:  
a 
Typical use failure rates may differ from those when used consistently and correctly. Use should be 
consistent with local regulations regarding the use of contraceptive methods for patients participating 
in clinical studies.  
b 
Hormonal contraception may be susceptible to interaction with the investigational product, which may 
reduce the efficacy of the contraceptive method. In this case, 2 highly effective methods of 
contraception should be utilized during the treatment period and for at least 5 weeks after the last 
dose of investigational product. 
 
Pregnancy Testing: 
x Any WOCBP should only be included after a negative pregnancy test. 
x Additional urine pregnancy testing should be performed at the Day 29 visit, and the 
Day 43 End-of-Study Follow-up visit. 
x Pregnancy testing will be performed whenever a menstrual cycle is missed or when 
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Collection of Pregnancy Information: 
x All occurrences of pregnancies must be reported on a Pregnancy Notification and 
Outcome Form (within 24 hours of awareness of any such pregnancy that occurs for 
either the study patient or for the female partner of a male patient); all pregnancies 
will be followed to outcome, which means until either the first well-baby visit for live 
births or until documented spontaneous or induced abortion. 
x While pregnancy itself is not considered to be an AE or SAE, any pregnancy 
complication or elective termination of a pregnancy will be reported as an AE or SAE. 
A spontaneous abortion is always considered to be an SAE and will be reported as 
such.  
x Pregnancies that occur beyond 90 days after the last dose of study drug will only 
need to be reported if there is an associated SAE that the Investigator assessed as 
related to study drug. 
x Safety Team will follow-up with the Investigator every 3 months regarding pregnancy 
outcome. The Investigator must follow the pregnancy to conclusion in order to 
determine the outcome. If the case is beyond 30 days from the expected due date 
and no information has been received regarding the outcome, the Safety Team will 
contact the Investigator to request outcome. 
Male Patients With Partners Who Become Pregnant  
x The Investigator will attempt to collect pregnancy information on any male patient’s 
female partner who becomes pregnant while the male patient is in this study. This 
applies only to male patients who receive BOS-589. 
x After obtaining the necessary signed informed consent from the pregnant female 
partner directly, the Investigator will record pregnancy information on the appropriate 
form and submit it to the sponsor as outlined above.  
Female Patients Who Become Pregnant  
x The Investigator will collect pregnancy information on any female patient who 
becomes pregnant while participating in this study. Information will be recorded on 
the appropriate form as noted above.  
x Any female patient who becomes pregnant while participating in the study will be 
withdrawn from the study. 
 
 


Boston Pharmaceuticals, Inc.
BOS-589-201
Protocol
Version 2.0
Confidential
Page 75 of 78
Appendix 3. The Bristol Stool Form Scale
(
)
CCI


Boston Pharmaceuticals, Inc. 
BOS-589-201 
Protocol 
Version 2.0 
 
Confidential 
 
Page 76 of 78 
Appendix 4. Concomitant Medications 
If the study site/investigator has any questions regarding permitted or excluded 
medications, the medical monitor should be contacted for further discussion. 
Medications permitted during the course of the study: 
x Any medication or vaccine (including over-the-counter or prescription medicines, 
vitamins, and/or supplements) that in the opinion of the Investigator, should not 
interfere with study procedures, compromise safety, or the scientific integrity of 
the data; 
x Stable doses of medications for allergies, migraines (with the exception of 
opioids for acute treatment), anxiety (e.g., as-needed use of benzodiazepines), 
depression or other chronic conditions at the discretion of the Investigator;  
x Stable doses of antidepressants. As-needed use of buspirone and 
benzodiazepines for anxiety; 
x Loperamide can be used during study as rescue medication based on 
protocol-specified guidelines (see Section 6.7.1) for allowable on-study rescue 
medications);  
x Acetaminophen, up to 2 g/day for up to 3 consecutive days; 
x The Investigator should monitor closely for potential side effects related to any of 
the following medicines which are considered moderate to highly sensitive 
substrates of CYP3A4. 
 
alprazolam  
aprepitant  
atorvastatin  
avanafil  
budesonide  
buspirone  
colchicine  
darifenacin  
dronedarone  
ebastine  
eletriptan  
felodipine  
lomitapide  
lovastatin  
midazolam  
naloxegol  
nisoldipine  
pimozide  
rivaroxaban  
sildenafil  
simvastatin  
tadalafil  
triazolam  
vardenafil  
 
Medications excluded during the course of the study: 


Boston Pharmaceuticals, Inc. 
BOS-589-201 
Protocol 
Version 2.0 
 
Confidential 
 
Page 77 of 78 
x 5-hydroxytriptamine (5-HT)3 or 5-HT4 receptor antagonists (e.g., alosetron); 
x Strong inhibitors of P-glycoprotein: 
amiodarone   
carvedilol  
clarithromycin  
dronedarone  
itraconazole  
propafenone  
quinidine  
ranolazine  
verapamil  
 
x Aspirin or aspirin-containing medications (> 325 mg of aspirin per day) or 
nonsteroidal anti-inflammatory drugs, when taken specifically for the symptoms 
of IBS; 
x Narcotic- or opioid-containing agents; 
x Cannabis-containing products; 
x Docusate; 
x Enemas; 
x GI preparations (including antacids containing aluminum or magnesium, 
antidiarrheal agents, antispasmodic agents, bismuth, peppermint oil, IBgard, 
FDgard, or prokinetic agents); 
x Planned use of rifaximin or oral antibiotics, with the exception of topical 
antibiotics or a 1-day course with an antibiotic); a patient will be allowed to 
remain in the study should unplanned use of antibiotics other than rifaximin occur 
after the patient has been randomized; 
x Any investigational drug. 
 
 


Boston Pharmaceuticals, Inc. 
BOS-589-201 
Protocol 
Version 2.0 
 
Confidential 
 
Page 78 of 78 
Appendix 5. 
Protocol Amendment History 
The Protocol Amendment Summary of Changes Table for the current amendment is 
provided as a separate document. 
DOCUMENT HISTORY 
Document 
Date of Issue 
Amendment 1, v.2.0 
09 May 2019 
Original, v.1.0 
25 January 2019 
 


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SAP_001.pdf: 
 
 
 
 
 
 
 
 
A Phase 2a, Randomized, Double-blind, Placebo-controlled, Multicenter 
Study to Evaluate the Efficacy, Safety, and Tolerability of BOS-589 in the 
Treatment of Patients with Diarrhea-predominant Irritable Bowel Syndrome 
(IBS-D) 
 
 
ClinicalTrials.gov Identifier: NCT03977155 
 
Statistical Analysis Plan, Version 1.2, dated 12-November-2020 
Statistical Analysis Plan, Version 1.1, dated 27-March-2020 
Statistical Analysis Plan, Version 1.0, dated 29-January-2020 
 
 


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