Clinical trial NCT04048135
Conditions: NASH
Keywords: NASH, NAFLD Liver diseases Fatty Liver Metabolic diseases
Sponsor: 89bio, Inc.
Start date: 2019-07-29; Completion date: 2022-01-19
Enrollment: 101 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
Part 1: This is a multi-center evaluation of pegozafermin (administered weekly or every other week) in a randomized, double-blind, placebo-controlled study administered for 12 weeks in participants with NASH and NAFLD at high risk of NASH, including a pre-defined number of participants with biopsy confirmed NASH and fibrosis stages F1-F3 to be enrolled.

Part 2: This is a multi-center, open label evaluation of pegozafermin at 27 mg administered weekly for 20 weeks in participants with biopsy-proven NASH (NAS ≥4, fibrosis stage F2 or F3).

Detailed description:

Arms: arm 1: Participants were administered 3 mg of pegozafermin QW, via subcutaneous (SC) injection, starting on Day 1 through Day 85. arm 2: Participants were administered 9 mg of pegozafermin QW, via SC injection, starting on Day 1 through Day 85. arm 3: Participants were administered 18 mg of pegozafermin QW, via SC injection, starting on Day 1 through Day 85. arm 4: Participants were administered 27 mg of pegozafermin QW, via SC injection, starting on Day 1 through Day 85. arm 5: Participants were administered 18 mg of pegozafermin Q2W, via SC injection, starting on Day 1 through Day 85. arm 6: Participants were administered 36 mg of pegozafermin Q2W, via SC injection, starting on Day 1 through Day 85. arm 7: Participants were administered placebo matching to pegozafermin QW or Q2W, via SC injection, starting on Day 1 through Day 85. arm 8: Participants were administered 27 mg of pegozafermin QW, via SC injection, starting on Day 1 through Day 134.
Interventions: intervention 1: Pegozafermin intervention 2: Placebo
Intervention descriptions: intervention 1: Subcutaneous injection intervention 2: Subcutaneous injection

Protocol documents:
Prot_000.pdf: 
CLINICAL STUDY PROTOCOL 
TITLE PAGE 
Title 
A Randomized, Double-Blind, Placebo-Controlled, Multiple 
Ascending Dose Study to Evaluate the Safety, Tolerability and 
Pharmacokinetic Properties of BIO89-100 Administered 
Subcutaneously in Subjects with Nonalcoholic Steatohepatitis 
(NASH) or with Nonalcoholic Fatty Liver Disease (NAFLD)and 
at High Risk of (NASH) 
Protocol Number:  
BIO89-100-002 
Compound Number: 
BIO89-100 
Study Phase:  
Phase 1b/2a 
Sponsor Name: 
89bio LTD. 
Legal Registered Address: 
6 Hamada Street, Herzliya 4676672 Israel 
Regulatory Agency 
Identifier Number: 
IND: 131934 
Sponsor Representative: 
 
Approval Date:  
11 March 2021 
Version: 
5.0 
Amendment: 
5.0 
This clinical study will be conducted in accordance with current Good Clinical Practice (GCP) 
as directed by the provisions of the International Council for Harmonisation (ICH); United 
States (US) Code of Federal Regulations (CFR), and European Union (EU) Directives and 
Regulations (as applicable in the region of the study); national country legislation; and the 
Sponsor’s Standard Operating Procedures (SOPs). 
CONFIDENTIALITY STATEMENT 
This document contains confidential and proprietary information (including confidential 
commercial information pursuant to 21CFR§20.61) and is a confidential communication of 
89bio LTD. The recipient agrees that no information contained herein may be published or 
disclosed without written approval from the Sponsor. If there is any actual or suspected 
breach of this obligation, the recipient agrees to promptly notify 89bio LTD. 
© 2020 89bio LTD. All rights reserved 
NCT #: NCT04048135


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SIGNATURE PAGE 
 
 
Sponsor Signatory: 
 
 
 
 
 
 89bio, Ltd. 
 
Date 
 
 
 
 
 
 
 
 
Medical Monitor Name and Contact Information will be provided separately. 


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PRINCIPAL INVESTIGATOR SIGNATURE PAGE 
Protocol Title 
A Randomized, Double-Blind, Placebo-Controlled, Multiple 
Ascending Dose Study to Evaluate the Safety, Tolerability and 
Pharmacokinetic Properties of BIO89-100 Administered 
Subcutaneously in Subjects with Nonalcoholic Steatohepatitis 
(NASH) or with Nonalcoholic Fatty Liver Disease 
(NAFLD)and at High Risk of (NASH)  
Protocol Number 
BIO89-100-002 
Version and Date 
5.0, 11 March 2021 
Amendment No.  
5.0 
IND Number 
IND: 131934 
 
 
I, the undersigned, have read this protocol and agree to personally supervise conduct of this 
protocol in accordance with ethical principles as outlined in the International Council for 
Harmonisation (ICH) guidelines on Good Clinical Practice, any applicable laws and 
requirements (including Part 54: Financial Disclosure by Clinical Investigators) and any 
additional conditions mandated by a regulatory authority and/or Institutional Review 
Board/Independent Ethics Committee (IRB/IEC). 
I acknowledge that I am responsible for the overall study conduct; I approve of and will 
comply with all conditions, instructions and restrictions described in this protocol. I am aware 
that my adherence to the above protocol is mandatory and that any changes in the protocol or 
consent form, except those necessary to eliminate apparent immediate hazards to human 
subjects, must first be approved in writing by 89bio, Ltd. And the respective IRB/IEC. 
I also agree that all information provided to me by the Sponsor, including this document, 
Investigator’s Brochure, case report form, and verbal and written information, will be kept 
strictly confidential and confined to the clinical personnel involved in conducting the study. It 
is recognized that this information may be related in confidence to the IRB/IEC. I also 
understand that reports of information about the study or its progress will not be provided to 
anyone not involved in the study other than to the Principal Investigator, or in confidence to the 
IRB/IEC or to the FDA or other legally constituted authority. 
 
 
 
Principal Investigator Signature 
Date 
 
 
Printed Name 
Institution 
City, Country 


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PROTOCOL AMENDMENT SUMMARY OF CHANGES 
DOCUMENT HISTORY 
Document 
Date 
Version 5.0, Amendment 5.0 
11 March 2021 
Version 4.1, Amendment 4.0 
22 October 2020 
Version 4.0, Amendment 4.0 
06 October 2020 
Version 3.0, Amendment 3.0 
25 March 2020 
Version 2.0, Amendment 2.0 
09 February 2020 
Version 1.1, Amendment 1.0 
29 June 2019 
Original, Version 1.0 
16 May 2019 
 
Amendment: 5.0, 11 March 2021 
Overall Rationale for Changes implemented in Version 5.0: 
Section # and Name 
Description of Change 
Brief Rationale 
Section 4.3.1 Part 2 
Inclusion Criteria 
IC # 3b: Revised to specify fasting plasma 
glucose ≥126 mg/dL or previous diagnosis 
of T2DM 
IC # 4b: Clarified the acceptable timeframe 
for historical liver biopsy 
IC # 5a: Clarified that MRI-PDFF will be 
adequate to assess steatosis if a Fibroscan 
model with CAP technology is unavailable 
IC # 7e and IC # 9: Clarified the definition 
of surgical sterilization and removed the 
requirement for medical documentation for 
eligibility consideration 
IC # 14: Added that COVID-19 protocols 
may be excepted with Medical Monitor (or 
designee) approval 
IC # 15: Removed the criterion  
To clarify the inclusion criteria 
and to facilitate enrolment in 
Part 2 of the study 


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Section # and Name 
Description of Change 
Brief Rationale 
Section 4.3.2 Part 2 
Exclusion Criteria 
Removed certain ECs: # 4 (cardiac 
arrhythmia), # 11 (major trauma or 
surgery), # 12 (acute illness), # 17 
(cigarette use), # 31 (blood donation)  
Clarified/revised the wording in the 
following ECs related to: (new #)   
Medical conditions: # 1, 2, 4, 6, 7, 8, 9, 10, 
12, and 13  
Prior and concomitant therapy: #15, 18, 
and 19 
Diagnostic assessments:  # 21, 22, 23, 24, 
and 25 
Other exclusions: 26, 27, and 31  
To clarify the exclusion 
criteria and to facilitate 
enrolment in Part 2 of the 
study 
Section 5.6.1 
Prohibited 
Medication 
Included gastric banding and sleeve 
surgery as prohibited bariatric surgery  
Removed insulin from the prohibited 
medication list 
Revised the wording on Vitamin E 
supplementation 
Specified that short term oral steroid bursts 
with/without taper as well as topical and 
inhaled steroids are allowed 
For consistency with the 
changes made in the exclusion 
criteria 
Appendix 4 
Contraceptives 
Removed the requirement for 
documentation of surgical sterilization 
 
For consistency with the 
changes made in IC # 7e and 
IC # 9 
Signature Page 
Changed the Sponsor Signatory 
Update in Sponsor Personnel 
 
 


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TABLE OF CONTENTS 
1.
PROTOCOL SUMMARY ..........................................................................11 
1.1. 
Synopsis ......................................................................................................11 
1.2. 
Schedule of Activities (SoA) ......................................................................20 
1.2.1. 
Part 1 ...........................................................................................................20 
1.2.2. 
Part 2 ...........................................................................................................31 
2.
INTRODUCTION ......................................................................................35 
2.1. 
Study Rationale ...........................................................................................35 
2.2. 
Background .................................................................................................35 
2.2.1. 
Non-alcoholic Steatohepatitis .....................................................................35 
2.2.2. 
FGF21 .........................................................................................................36 
2.2.3. 
BIO89-100 ..................................................................................................37 
2.3. 
Benefit / Risk Assessment ..........................................................................37 
2.3.1. 
Risk Assessment .........................................................................................37 
2.3.1.1. 
Potential Risk: Immunogenicity .................................................................38 
2.3.1.2. 
Potential Risk: Bone Effects .......................................................................39 
2.3.1.3. 
Potential Risk: Female Reproductive Effects .............................................39 
2.3.1.4. 
Potential Risk: Injection Site Reactions .....................................................40 
2.3.1.5. 
Potential Risk: Gastrointestinal Effects ......................................................40 
2.3.1.6. 
Potential Risk of Liver Biopsy ...................................................................41 
2.3.2. 
Benefit Assessment .....................................................................................41 
2.3.3. 
Update: Preliminary Data from Part 1 ........................................................41 
2.3.4. 
Overall Benefit Risk Assessment ...............................................................42 
3.
PART 1 .......................................................................................................43 
3.1. 
Part 1 Objectives and Endpoints .................................................................43 
3.2. 
Part 1 Study Design ....................................................................................45 
3.2.1. 
Scientific Rationale for Study Design ........................................................46 
3.2.2. 
Rationale for Study Population ...................................................................47 
3.2.3. 
Justification of Treatment Duration ............................................................47 


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3.2.4. 
Justification for Dose ..................................................................................48 
3.2.4.1. 
Justification for Weekly (QW) and Once Every Two Weeks (Q2W) 
Dosing Regimens ........................................................................................49 
3.2.5. 
End of Study Definition ..............................................................................49 
3.3. 
Part 1 Eligibility Criteria ............................................................................49 
3.3.1. 
Part 1 Inclusion Criteria ..............................................................................49 
3.3.2. 
Part 1 Exclusion Criteria .............................................................................51 
4. 
PART 2 .......................................................................................................57 
4.1. 
Part 2 Objectives and Endpoints .................................................................57 
4.2. 
Part 2 Study Design ....................................................................................60 
4.2.1. 
Scientific Rationale for Part 2 Study Design ..............................................60 
4.2.2. 
Rationale for Part 2 Study Population ........................................................61 
4.2.3. 
Justification for Part 2 Treatment Duration ................................................61 
4.2.4. 
Justification for Part 2 Dose .......................................................................61 
4.2.5. 
Part 2 End of Study Definition ...................................................................62 
4.3. 
Part 2 Eligibility Criteria ............................................................................62 
4.3.1. 
Part 2 Inclusion Criteria ..............................................................................62 
4.3.2. 
Part 2 Exclusion Criteria .............................................................................65 
5. 
STUDY INTERVENTION ........................................................................69 
5.1. 
Study Intervention(s) Administered ...........................................................69 
5.2. 
Administration Instructions ........................................................................70 
5.3. 
Preparation/Handling/Storage/Accountability ............................................70 
5.4. 
Measures to Minimize Bias: Randomization ..............................................70 
5.5. 
Study Intervention Compliance ..................................................................71 
5.6. 
Concomitant Therapy .................................................................................71 
5.6.1. 
Prohibited Medications/Therapies ..............................................................72 
5.7. 
Stopping Rules and Dose Modifications ....................................................72 
5.7.1. 
Monitoring and Discontinuation for Suspected Drug-induced Liver Injury 
(DILI) ..........................................................................................................72 
5.8. 
Screen Failures ............................................................................................75 
5.9. 
Intervention after the End of Study ............................................................75 
6. 
DISCONTINUATION OF STUDY INTERVENTION AND SUBJECT 
DISCONTINUATION/WITHDRAWAL ..................................................76 


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6.1. 
Discontinuation of Study Intervention ........................................................76 
6.2. 
Subject Discontinuation/Withdrawal from the Study .................................76 
6.3. 
Lost to Follow-up .......................................................................................76 
7. 
STUDY ASSESSMENTS AND PROCEDURES ......................................78 
7.1. 
Efficacy Assessments .................................................................................78 
7.1.1. 
Liver Biopsy and Scoring (Part 2 Only) .....................................................78 
7.2. 
Safety Assessments .....................................................................................79 
7.2.1. 
Physical Examinations ................................................................................79 
7.2.2. 
Vital Signs ..................................................................................................79 
7.2.3. 
Electrocardiograms .....................................................................................79 
7.2.4. 
Columbia-Suicide Severity Rating Scale (C-SSRS) ...................................79 
7.2.5. 
Clinical Safety Laboratory Assessments ....................................................80 
7.3. 
Adverse Events and Serious Adverse Events .............................................80 
7.3.1. 
Time Period and Frequency for Collecting AE and SAE Information .......81 
7.3.2. 
Method of Detecting AEs and SAEs ..........................................................81 
7.3.3. 
Follow-up of AEs and SAEs .......................................................................81 
7.3.4. 
Regulatory Reporting Requirements for SAEs ...........................................81 
7.3.5. 
Pregnancy ...................................................................................................82 
7.4. 
Treatment of Overdose ...............................................................................82 
7.5. 
Pharmacokinetics ........................................................................................83 
7.6. 
Pharmacodynamics and Biomarkers ...........................................................83 
7.6.1. 
Immunogenicity Assessments ....................................................................83 
7.6.2. 
Pharmacodynamics .....................................................................................84 
7.6.2.2. 
Homeostatic Model Assessment for Insulin Resistance (HOMA-IR; Part 1 
only) ............................................................................................................85 
7.6.2.4. 
N-terminal Propeptide of Type III Collagen (Pro-C3) ...............................86 
7.6.2.5. 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction 
(MRI-PDFF) ...............................................................................................86 
7.6.3. 
Exploratory Analyses ..................................................................................87 


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8. 
STATISTICAL CONSIDERATIONS .......................................................88 
8.1. 
Statistical Hypotheses .................................................................................88 
8.2. 
Sample Size Determination ........................................................................88 
8.3. 
Populations for Analyses ............................................................................89 
8.4. 
Statistical Analyses .....................................................................................89 
8.4.1. 
Safety and Tolerability Analyses ................................................................90 
8.4.2. 
PK Analyses ................................................................................................91 
8.4.3. 
Immunogenicity Analyses ..........................................................................91 
8.5. 
Interim Analyses .........................................................................................92 
8.6. 
Safety Monitoring Committee (SMC) ........................................................92 
9. 
SUPPORTING DOCUMENTATION AND OPERATIONAL 
CONSIDERATIONS ..................................................................................93 
9.1. 
Appendix 1: Regulatory, Ethical, and Study Oversight Considerations ....93 
9.1.1. 
Regulatory and Ethical Considerations ......................................................93 
9.1.2. 
Financial Disclosure ...................................................................................93 
9.1.3. 
Informed Consent Process ..........................................................................93 
9.1.4. 
Data Protection ...........................................................................................94 
9.1.5. 
Dissemination of Clinical Study Data ........................................................94 
9.1.6. 
Data Quality Assurance ..............................................................................94 
9.1.7. 
Source Documents ......................................................................................95 
9.1.8. 
Study and Site Closure ................................................................................95 
9.1.9. 
Publication Policy .......................................................................................96 
9.2. 
Appendix 2: Clinical Laboratory Tests .......................................................97 
9.3. 
Appendix 3: Adverse Events: Definitions and Procedures for Recording, 
Evaluating, Follow-up, and Reporting ........................................................99 
9.3.1. 
Definition of AE .........................................................................................99 
9.3.2. 
Definition of Suspected and Unsuspected Adverse Reaction ...................100 
9.3.3. 
Definition of Events to Monitor ...............................................................100 
9.3.4. 
Definition of SAE .....................................................................................100 
9.3.5. 
Recording and Follow-Up of AE and/or SAE ..........................................101 
9.3.6. 
Reporting of SAEs ....................................................................................103 


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9.4. 
Appendix 4: Contraceptive Guidance and Collection of Pregnancy 
Information ...............................................................................................104 
9.5. 
Appendix 5: Considerations in Response to Coronavirus Disease 2019 
(COVID-19) Pandemic .............................................................................107 
9.6. 
Appendix 6: Glossary ...............................................................................109 
10. 
REFERENCES .........................................................................................121 
LIST OF TABLES 
Table 1 
Study Cohorts .............................................................................................18 
Table 2 
Part 1 Schedule of Activities for Cohorts 1-4 .............................................20 
Table 3 
Part 1 PK Sample Collection for Cohorts 1-4 – QW Dosing Interval ........24 
Table 4 
Part 1 Schedule of Activities for Cohorts 5 and 6 ......................................25 
Table 5 
Part 1 PK Sample Collection for Cohorts 5 and 6 – Q2W Dosing Interval
 ....................................................................................................................30 
Table 6 
Schedule of Activities for Part 2 (Cohort 7) ...............................................31 
Table 7 
Discontinuation Criteria in Subjects with Abnormal Baseline ALT or AST 
Values .........................................................................................................74 
Table 8 
Adverse Event Reporting Periods ...............................................................81 
Table 9 
Protocol-Required Laboratory Assessments ...............................................97 
 


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1. 
PROTOCOL SUMMARY 
1.1. 
Synopsis 
Protocol title: A Randomized, Double-Blind, Placebo-Controlled, Multiple Ascending Dose 
Study to Evaluate the Safety, Tolerability and Pharmacokinetic Properties of BIO89-100 
Administered Subcutaneously in Subjects with Nonalcoholic Steatohepatitis (NASH) or with 
Nonalcoholic Fatty Liver Disease (NAFLD) and at High Risk of NASH 
Rationale 
BIO89-100, developed by 89bio, is a glycoPEGylated analogue of fibroblast growth factor 21 
(FGF21), a metabolic hormone secreted by the liver, adipose tissue, skeletal muscle, and the 
pancreas, that is regulated by nutritional status and affects energy expenditure, and glucose and 
lipid metabolism (Lee, 2014) (Arner, 2008) (Park, 2016). Administration of exogenous FGF21 
is being explored as a method to treat obesity-associated insulin-resistance disorders, a 
category that includes Non-Alcoholic Fatty Liver Disease (NAFLD), the most common chronic 
liver disorder worldwide, and its higher-risk variant Non-Alcoholic Steatohepatitis (NASH). 
There are currently no approved pharmaceutical treatments for NASH (Friedman, 2018). 
Several nonclinical and clinical studies have shown that administration of various FGF21 
analogues had beneficial effects on serum lipid and insulin-resistance as well as on liver fat 
(Zhang 2014) (Gaich, 2013) (Sanyal, 2019). 
This Phase 1b/2a study has 2 parts. Part 1 is designed to assess the safety and tolerability of 
repeated subcutaneous (SC) escalating doses of BIO89-100 in subjects with a diagnosis of 
NASH, or who have NAFLD with a high risk of NASH. Part 2 is open-label, designed to 
assess the effect of repeated SC doses of one dose level of BIO89-100, administered for 
20 weeks, on liver histology in subjects with biopsy-proven NASH with NAFLD Activity 
Score (NAS) ≥4, fibrosis stage F1 with high risk, F2 or F3. 
The study includes safety, pharmacokinetic (PK), pharmacodynamic (PD), and 
immunogenicity characterizations to allow a preliminary assessment of a safe and efficacious 
dose range to be studied in further clinical studies. 
Rationale for Part 2 (Cohort 7) 
To obtain an initial understanding of the potential histological benefit of BIO89-100, an 
open-label cohort (Cohort 7) has been added under Part 2 for evaluation of the effect of 
20 weeks of dosing with BIO89-100 at a dose of 27 mg/week (QW) on liver histological 
endpoints. Assessments for this cohort will include safety, tolerability, PK, and PD parameters 
(with some modifications in comparison to Cohorts 1-6, see Cohort 7 Schedule of Activities 
[SoA], Table 6). This cohort will enroll 20 subjects with biopsy-proven NASH (NAS ≥4, 
fibrosis stage F1 with high risk, F2 or F3) as determined by a baseline biopsy (to be done 
during screening; subjects with historical biopsies performed within 24 weeks of Day 1, that 
are available for central read and determined to meet inclusion criteria by the central reader, 
will also be eligible). Subjects will undergo a second liver biopsy within 
 after the last 
dose of study intervention. Histological assessment will include ≥2-point improvement in NAS 


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with at least a 1-point improvement in ballooning or lobular inflammation, and no worsening of 
fibrosis  (primary endpoint); improvement of fibrosis ≥1 stage without worsening of NAS 
(secondary endpoint); and NASH resolution without worsening of fibrosis0F1 (secondary 
endpoint). Additional exploratory histological endpoints (e.g., other endpoints related to NAS, 
fibrosis, or both; histological assessments in subgroups; e.g., subjects with ≥30% relative 
reduction in hepatic fat as assessed by magnetic resonance imaging [MRI]-proton density fat 
fraction [PDFF] on Week 12 compared to baseline) will be evaluated. Biopsies will be assessed 
by a central reader; in addition, biopsies will be assessed with artificial intelligence (AI)-based 
technology.  
Objectives and Endpoints 
Part 1 (Cohorts 1-6) 
Objectives 
Endpoints 
Primary 
• To evaluate the safety and 
tolerability of ascending 
multiple SC injections of 
BIO89-100 in subjects with 
NASH or who have NAFLD 
and at a high risk of NASH 
• Frequency and severity of adverse 
events (AEs) and serious adverse 
events (SAEs) 
• Number of subjects who discontinued 
due to AEs and due to related AEs 
• To characterize BIO89-100 
PK 
• Maximal observed serum 
concentrations (Cmax) within a dosing 
interval 
• Area under the serum drug 
concentration by time curve within a 
dosing interval (AUC0-tau) 
• Time to achieve Cmax (tmax) 
• Terminal elimination half-life (t½) 
Additional PK parameters may be calculated if 
deemed appropriate.  
The serum concentration-time data may also be 
used for population PK modeling with the results 
reported separately from the clinical study 
report. 
Secondary 
 
1 Resolution of NASH includes the total absence of ballooning (score=0) and absent or mild inflammation (score 0 
to 1) associated with at least a 2 point reduction in NAS, and no worsening if fibrosis (progression ≥ 1 stage). 


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Objectives 
Endpoints 
• To evaluate the 
immunogenicity of BIO89-
100 as measured by presence 
of anti-drug antibodies (ADA) 
• Assessment of the incidence and 
characteristics of ADA after dosing 
(e.g., titer and/or binding specificity, 
to the FGF21, PEG part of BIO89-
100, and neutralizing 
immunogenicity) 
• Impact of the presence of ADAs on 
serum BIO89-100 concentrations and 
clinical safety 
Additional ADA assessments will be performed 
if a severe hypersensitivity reaction (e.g., 
anaphylaxis) is observed. 
• To characterize biomarkers, 
PD profile, and biological 
activity of BIO89-100 
administered at ascending 
doses and with both QW and 
Q2W dosing intervals 
• To evaluate the time, dose, 
and exposure relationship of 
BIO89-100 on biological 
activity, as assessed by 
biomarkers and PD 
Change and percentage change from baseline in 
the following biomarkers/PD parameters: 
• Anthropomorphic measurements: 
− Body weight 
• Laboratory parameters: 
− Triglycerides 
− Non-high density lipoprotein (non-
HDL) cholesterol 
− High density lipoprotein (HDL-c) 
− Low density lipoprotein (LDL-c) 
− Hemoglobin A1c (HbA1c) 
− Homeostatic model assessment of 
insulin resistance (HOMA-IR) 
− Liver function tests: alanine 
transaminase (ALT), aspartate 
transaminase (AST)  
− Adiponectin 
− N-terminal propeptide of type III 
collagen (Pro-C3) 
− Free fatty acids and Adipo-IR (fasting 
free fatty acids  fasting insulin) 
• Imaging measures: 


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Objectives 
Endpoints 
− Magnetic resonance imaging – Proton 
density fat fraction (MRI-PDFF) 
Other Safety Endpoints 
 
Part 2 (Cohort 7) 
Objectives 
Endpoints 
Primary 
• To evaluate the safety and 
tolerability of SC injections of 
27 mg BIO89-100, 
administered weekly for 20 
weeks, in subjects with 
biopsy-proven NASH (NAS 
≥4, fibrosis stage F1 with high 
risk, F2 or F3) 
• Frequency of AEs and SAEs 
• To characterize effect of 
BIO89-100 on liver histology 
• At least a 2-point improvement in 
NAFLD Activity Score (NAS) with 
at least a 1-point improvement in 
ballooning or lobular inflammation, 
and no worsening of fibrosis 
Secondary 
• To characterize biomarkers, 
PD profile, and biological 
activity of BIO89-100 
Change and percentage change from baseline in 
the following biomarkers/PD parameters: 
• Anthropomorphic measurements: 
− Body weight 
• Laboratory parameters: 
− Triglycerides 


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Objectives 
Endpoints 
− non-HDL cholesterol 
− HDL-c 
− LDL-c 
− HbA1c 
− Liver function tests: ALT, AST 
− Pro-C3 
• Imaging measures: 
− MRI-PDFF   
• To characterize effect of 
BIO89-100 on liver histology 
• Improvement of fibrosis ≥1 stage 
without worsening of NASH 
• NASH resolution without worsening 
of fibrosis2 
• To characterize BIO89-100 
PK 
• Trough concentration of BIO89-100 
Other Safety Endpoints 
 
2 Resolution of NASH includes the total absence of ballooning (score=0) and absent or mild inflammation (score 0 
to 1) associated with at least a 2-point reduction in NAS, and no worsening if fibrosis (progression ≥ 1 stage). 


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Objectives 
Endpoints 
Overall Design 
This study has 2 parts: 
• Part 1 is a randomized, double-blind, placebo-controlled, multiple ascending dose 
(MAD) study to evaluate the safety, tolerability, PK and PD profiles, and 
immunogenicity of BIO89-100 administered SC in approximately 83 subjects with 
NASH, or with NAFLD who are at a high risk of NASH. This multi-site study will 
consist of 6 cohorts and will evaluate 2 dosing schedules, weekly (QW; Cohorts 1 to 
4) and every 2 weeks (Q2W; Cohorts 5 and 6) (Table 1). 
• Part 2 is an open-label cohort (Cohort 7) in which BIO89-100 will be administered 
SC weekly at a single dose level (27 mg QW) to 20 subjects with biopsy-proven 
NASH (NAS ≥4, fibrosis stage F1 with high risk, F2 or F3), to evaluate effect on 
histological endpoints.  
The study will include a Screening period, a Treatment period, and a Follow-up period. After 
signing informed consent, subjects will undergo screening assessments to determine eligibility 
over a period of up to 60 days. 
Part 1 (Cohorts 1-6) 
Eligible subjects for Cohorts 1-6 will be randomized for each cohort as described in 
Section 5.4. Randomization to cohorts 3 and 5 will be stratified by biopsy-confirmed NASH 
with fibrosis status F1, F2, or F3 as described in Section 5.4.The decision to randomize into the 
next cohort (dose level) will be approved by a Safety Monitoring Committee (SMC), 
comprised of Principal Investigator participating in the study, the Clinical Research 
Organization (CRO) Medical Monitor, and the Sponsor Medical Monitor. The SMC will 
review blinded safety data (adverse events, clinical laboratory data, vital signs, and 
electrocardiograms [ECG]), summary of PK data, if available, and other blinded relevant data. 
There will be 2 dose escalation decisions. After Cohort 1 completes the Day 36 visit, the SMC 
will decide whether subjects can be randomized into Cohorts 2 and 5 (both cohorts to start 
concurrently). After at least 8 subjects from both Cohort 2 and Cohort 5, including at least 
1 subject on placebo in each cohort, complete the Day 36 visit, the SMC will decide whether 
subjects can be randomized into Cohorts 3, 4, and 6 (all three cohorts to start concurrently). 


Protocol BIO89-100-002 
 
 
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For all subjects, treatment with study intervention may be continued for 8 additional weeks 
(total of 12 weeks) following review of individual safety data (AEs, ECG, physical 
examination, and safety laboratory) through Day 30 (4 weeks) by the site Investigator, CRO 
Medical Monitor and Sponsor Medical Monitor. Subjects who experience clinically significant 
treatment-emergent AEs that are assessed as a potential risk to subject safety will be 
discontinued from study intervention. The decision to discontinue study intervention will be 
made by the site Investigator following discussion with the Sponsor Medical Monitor. 
Cohorts 1 to 4 (weekly regimen): On Day -1, eligible subjects will be randomized (as described 
above) and be treated with weekly (QW) SC injection of study intervention starting on Day 1 
and continuing through Day 85; subjects will be domiciled at the Phase 1 unit from Day -1 (day 
before 1st dose) to Day 2 and from Day 28 (day before the 5th dose) to Day 30. Subjects will 
attend ambulatory clinic visits for dosing and evaluations per the Schedule of Activities (SoA; 
Section 1.2.1, Table 2). On the Day 8, Day 15, and Day 22 ambulatory dosing visits, subjects 
will remain at the study site for observation for at least 2 hours post dose. 
Cohorts 5 and 6 (every 2 weeks regimen): On Day -1, eligible subjects will be randomized (as 
described above) and be treated with SC injection of study intervention every 2 weeks (Q2W) 
starting on Day 1 and continuing through Day 85; subjects will be domiciled at the Phase 1 unit 
from Day -1 (day before 1st dose) to Day 2 and from Day 28 (day before the 3rd dose) to 
Day 30. Subjects will attend ambulatory clinic visits for dosing and evaluations per the SOA 
(Section 1.2.1, Table 4). On the Day 15 ambulatory dosing visit, subjects will remain at the 
study site for observation for at least 2 hours post dose. On Days 64 and 78, subjects will be 
contacted by phone to inquire about AEs and concomitant medications. 
Subjects in all cohorts will be followed up on Day 92 (1 week post last dose of study 
intervention) and Day 113, 4 weeks post last dose of study intervention (End of Study visit). 
Subjects who are found to be neutralizing anti-drug antibodies (NAb)-positive at the End of 
Study visit will be followed for 3-5 months after this visit or until stable or declining as deemed 
by the Medical Monitor. 
Part 2 (Cohort 7) 
Part 2 is an open-label cohort that will enroll 20 subjects with biopsy-proven NASH and 
fibrosis (NAS ≥4, fibrosis stage F1 with high risk, F2 or F3). Subjects will either undergo a 
liver biopsy during screening or have a recent liver biopsy (within ≤24 weeks before Day 1, 
available to be evaluated for eligibility by the central reader) that meets study inclusion criteria. 
Eligible subjects will be treated with weekly (QW) SC injection of study intervention starting 
on Day 1 and continuing through Day 134 (20 weeks of treatment). Subjects will attend 
ambulatory clinic visits for dosing and evaluations and home visits for dosing as per the 
Schedule of Activities3 (SoA; Section 1.2.2, Table 6). Subjects will undergo a second liver 
biopsy within 14 days after the last dose of study intervention. Subjects will be followed up on 
Day 141 (1 week post last dose of study intervention) and Day 162 (4 weeks post last dose of 
study intervention, End of Study visit). 
 
 
3 If home administration is not feasible, not desired by the subject, or deemed inappropriate option by the 
investigator for any reason, in-clinic study intervention administration may take place at some or all timepoints. 


Protocol BIO89-100-002 
 
 
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. More frequent 
testing (e.g., every month) or testing for a longer period of time may be requested in the event 
of safety-related concerns.  
Number of Subjects and Intervention Groups 
A total of approximately 103 subjects, consisting of: 6 subjects to receive active and 2 subjects 
to receive placebo in Cohort 1; 12 subjects to receive active and 3 subjects to receive placebo 
in Cohort 2; 9 subjects to receive active and 3 subjects to receive placebo in Cohorts 4 and 6; 
14 subjects to receive active and 4 subjects to receive placebo in Cohorts 3 and 5; and 
20 subjects to receive active (open-label) in Cohort 7. Dose escalation cohorts are shown in 
Table 1. Based on emerging data and SMC recommendation, the Sponsor can decide to modify 
the dose of any subsequent cohort. 
Table 1 
Study Cohorts 
Cohort 
Dose 
Levela 
Frequency and Route of Administration 
Number of Subjects 
BIO89-100 
Placebo 
1 
3 mg 
Weekly (QW), SC to abdomen (1 injection) 
Days 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, 78, 
and 85 
Total doses (initial 4 weeks + 8-week 
extension): 5 + 8 
6 
2 
2 
9 mg 
QW, SC to abdomen (1 injection) 
Days 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, 78, 
and 85 
Total doses (initial 4 weeks + 8-week 
extension): 5 + 8 
12 
3 
3 
18 mg 
QW, SC to abdomen (1 injection) 
Days 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, 78, 
and 85 
Total doses (initial 4 weeks + 8-week 
extension): 5 + 8 
14 
4 
4 
27mg 
QW, SC to abdomen (2 injections) 
Days 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, 78, 
and 85 
Total doses (initial 4 weeks + 8-week 
extension): 5 + 8 
9 
3 
5 
18 mg 
Every 2 weeks (Q2W), SC to abdomen 
(1 injection) 
Days 1, 15, 29, 43, 57, 71, and 85. 
Total doses (initial 4 weeks + 8-week 
extension): 3 + 4 
14 
4 


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Cohort 
Dose 
Levela 
Frequency and Route of Administration 
Number of Subjects 
BIO89-100 
Placebo 
6 
36 mg 
Q2W, SC to abdomen (2 injections) 
Days 1, 15, 29, 43, 57, 71, and 85. 
Total doses (initial 4 weeks + 8-week 
extension): 3 + 4 
9 
3  
7 (open-
label) 
27 mg 
QW, SC to abdomen (2 injections) 
Days 1, 8, 15, 22, 29, 36, 43, 50, 57, 64, 71, 78, 
85, 92, 99, 106, 113, 120, 127, and 134 
Total doses: 20 
20 
NA 
a The actual doses will be ±5% the mg dose due to technical considerations related to drug withdrawal from the 
vials into the syringes for injection. This difference is considered negligible for subject exposure. 
Study Duration 
Part 1 (Cohorts 1-6) 
For each subject in Part 1, the total duration of study participation will be up to 172 days 
(24.5 weeks), excluding the potential study interruption due to the COVID-19 pandemic: 
Screening period: 
≤ 60 days (8.5 weeks) 
Treatment period: 
84 days (12 weeks) 
Follow-up period 
28 days (4 weeks) 
Part 2 (Cohort 7) 
For each subject in Part 2 (Cohort 7), the total duration of study participation will be up to 
221 days (31.5 weeks), excluding the potential study interruption due to the COVID-19 
pandemic: 
Screening period: 
≤ 60 days (8.5 weeks) 
Treatment period: 
134 days (20 doses in 19 weeks) 
Follow-up period 
28 days (4 weeks 4) 
Safety Monitoring Committee: Yes (Cohorts 1-6) 
 
4 Subjects who are found to be NAb-positive at the End of Study visit will be followed every 3 months starting 
from when the site has been notified of the positive result, until: (1) neutralizing antibodies are no longer 
detectable or (2) the subject has been followed for a period of at least 1 year (± 4 weeks). More frequent testing 
(e.g., every month) or testing for a longer period of time may be requested in the event of safety-related concerns. 


Protocol BIO89-100-002 
 
 
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1.2. 
Schedule of Activities (SoA) 
1.2.1. 
Part 1 
Table 2 
Part 1 Schedule of Activities for Cohorts 1-4 
Cohort 1-4 
Assessments 
Screening Period 
Treatment Period 
 
FU 
Period 
Study Visit 
Study Day 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical history/demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Percutaneous liver biopsy 
(optional)b 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Prior medications 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion and exclusion criteria 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Complete physical examc 
X 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
X  
X 
Symptom-directed physical 
examd 
 
 
X 
B 
P 
 
 
 B B B 
 
B 
 
 
 
 
B 
 
 
Body weighte 
X 
 
 
B 
 
 
 
 B B B 
 
B 
 
 
 
 
B 
 
 
Waist and hip measurements 
X 
 
 
B 
 
 
 
 
 
 
 
 
B 
 
 
 
 
 
X 
X 
Fibroscanf 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
12-lead ECG (single)g 
X 
 
 
B 
X 
 
 
 B B B 
 
B 
X 
 
 
 
B 
X 
X 
Urine drug screen and, alcohol 
breath test 
X 
 
Xh 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical laboratory testsi 
X 
X 
 
B 
X 
 
 
 B B B 
 
B 
X 
 
 
 
B 
X 
X 
Cortisol (24 hour urine 
collection)j 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
Urinalysis 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
X 
X 
HbA1C 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
Serologyk 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
TSH 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Cohort 1-4 
Assessments 
Screening Period 
Treatment Period 
 
FU 
Period 
Study Visit 
Study Day 
Pregnancy test in WOCBP onlyl 
X(S) 
 
X 
 
 
 
 
 B B B 
 
B 
 
 
 
 
B 
B 
X 
X 
Vital signsm 
X 
 
X 
X 
P X X X X X X X 
X 
P 
X 
X 
X 
X 
X 
X 
X 
Columbia-Suicide Severity 
Rating Scale (C-SSRS) 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
D50 
 
X 
 
Randomization 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Study Intervention 
administrationn 
 
 
 
X 
 
 
 
 X X X 
 
X 
 
 
 
 
X 
X 
 
 
PK blood collectiono 
 
 
 
X 
X X X X B B B 
 
X 
X 
X 
X 
X 
B 
X 
 
PD and biomarker blood 
sampling: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
IGF-1, total 
 
 
 
B 
 
 
 
 B B 
 
 
B 
 
 
 
 
 
X 
 
Adiponectin 
 
 
 
B 
 
 
 
 B 
 
 
 
B 
 
 
 
 
 
X 
X 
CK-18, ELF panel and Pro-C3 
 
 
 
B 
 
 
 
 
 
 
 
 
B 
 
 
 
 
 
X 
Free fatty acid 
 
 
 
B 
 
 
 
 B 
 
 
 
B 
 
 
 
 
 
X 
X 
Oral glucose tolerance test 
(OGTT)p  
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
Insulin (not part of OGTT) 
 
 
 
B 
 
 
 
 B 
 
 
 
B 
 
 
 
 
 
 
X 
HOMA-IR calculation 
 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
MRI-PDFF; visceral fat; and 
subcutaneous fatq 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
Plasma sample for potential 
future bone biomarkers analysisr 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
Pharmacogenomic (DNA) blood 
sampling 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Exploratory biomarker analysiss  
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
Immunogenicity t 
 
 
 
B 
 
 
 
 
 
B 
 
 
B 
 
 
 
 
 
Xu 
X 
Endogenous FGF21v 
 
 
 
B 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Protocol BIO89-100-002 
 
 
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Cohort 1-4 
Assessments 
Screening Period 
Treatment Period 
 
FU 
Period 
Study Visit 
Study Day 
Adverse event monitoringw 
X 
X 
X 
←-===========================================→ 
X 
X 
Concomitant medication  
X 
X 
X 
←-===========================================→ 
X 
X 
Domiciledx 
 
 
X 
X 
X 
 
 
 
 
 
 
X 
X 
X 
 
 
 
 
 
 
 
Abbreviations: B = predose; D = Day; CK = cytokeratin; DNA = deoxyribonucleic acid; ECG=electrocardiogram; ELF = enhanced liver fibrosis; EOS = end of study; 
ET= early termination; FGF21 = fibroblast growth factor 21; FU = Follow-Up; HbA1c=hemoglobin A1c; IGF-1 = insulin-like growth factor; MRI-PDFF = magnetic 
resonance imaging based proton density fat fraction; P = Pre-discharge; PK=pharmacokinetic; Pro-C3 = N‐terminal propeptide of type III collagen; S = serum; TSH = 
thyroid stimulating hormone; WOCBP=women of child-bearing potential. 
Table 2 footnotes 
All outpatient visits will have a study window of ±1 day. 
a. 
For any subject who withdraws before completion of the study, an Early Termination (ET) visit will be conducted, if possible; with the same assessments 
as the Day 92 visit.  
b. Optional liver biopsy may be performed  for subjects who do not have a medical contraindication to undergoing a liver biopsy, and did not have a liver 
biopsy in the 24 months prior to screening.|For subjects who underwent a previous liver biopsy >24 months prior to screening, that would not have 
qualified for the study based on results of the initial biopsy, there should be a sound clinical basis to expect different findings in a repeat biopsy based on 
medical judgment. Any biopsy will need to be approved in advance by Sponsor and only after the subject has met all other inclusion and exclusion 
criteria,  
c. 
Complete physical exam done at screening to include recording height, weight, and calculating BMI. 
d. Symptom-directed physical exam will be done pre-dose and prior to discharge from the Phase 1 unit on domiciled dosing visits (1st and 5th dose), and pre-
dose on ambulatory dosing visits. Additional physical examinations will be performed if clinically indicated. 
e. 
On domiciled and ambulatory visits, body weight will be measured pre-dose. 
f. 
Fibroscan should be performed during screening, prior to magnetic resonance imaging (MRI) for all subjects. 
g. 12-lead safety ECGs will be recorded as single bedside measurements; on domiciled dosing visits (1st and 5th dose), ECG will be measured pre-dose and 
24 hours post-dose. On ambulatory dosing visits, ECG will be measured pre-dose. Additional ECG may be conducted if clinically indicated. 
h. On Day -1, urine drug screen can be done at a local laboratory, however, a sample should also be collected for central laboratory evaluation. 
i. 
Clinical laboratory tests will include biochemistry, hematology, and coagulation, and FSH for determination of post-menopausal status ; on domiciled 
dosing visits (1st and 5th dose), clinical laboratory will be collected pre-dose and 24 hours post dose; on ambulatory visits, clinical laboratory will be 
collected pre-dose. For all subjects, alanine transaminase (ALT) and aspartate transaminase (AST) will be collected twice during the Screening period (at 
least 2 weeks apart), with the 2nd assessment to be collected at Day -7 to Day -3. A 3rd assessment, if required, will be collected via unscheduled visit 
(refer to Exclusion criterion 3). 
j. 
Ambulatory 24-hour urine collection for cortisol to be done within 14 days from baseline, on Day 50 and Day 92/ET. It is recommended that subjects 
collect urine for the 24 hours prior to D-7 to-3 visit and bring the container with them to this visit. Alternatively, urine can be collected for the 24 hours 


Protocol BIO89-100-002 
 
 
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prior to the randomization visit (D-1) and brought to the site on D-1 (the result is not required for eligibility confirmation).  For Day 50 and D92/ET – 
subject will start collection 24 hours before coming into the clinic and bring the sample to the visit. 
k. Serology tests will include Hepatitis B surface antigen, Hepatitis C Virus, and Human Immunodeficiency Virus (HIV) 1 and 2 antibodies. 
l. 
Serum urine pregnancy test will be conducted at screening; at all other timepoints urine pregnancy test will be done to guide clinical decisions to dose on 
dosing days. Prior to the 1st dosing, the baseline urine pregnancy test will be performed on D-1 to allow for randomization on that day. If urine test is 
positive, a confirmatory serum pregnancy test will be conducted. 
m. k Vital signs include supine blood pressure, pulse, body temperature, and respiratory rate; vital signs will be measured pre-dose (prior to scheduled blood 
draws and study intervention administration); 1, 12 and 24 hours post-dose and prior to discharge on domiciled dosing visits (1st and 5th doses); on dosing 
ambulatory visits, vital signs will be measured pre-dose (prior to scheduled blood draws and study intervention administration) and prior to discharge; on 
non-dosing visits, vital signs will be measured prior to scheduled blood draws. Starting from randomization, blood pressure and pulse will be measured in 
duplicate, the first measurement will be taken up to 15 minutes before the indicated timepoint. Additional vital signs measurement may be done if 
clinically indicated. Subjects must be in a supine or semi-erect/seated position and resting for at least 5 minutes prior to measurements. 
n. Study intervention will be administered SC to the abdomen region by qualified study personnel. 
o. PK blood samples will be collected as shown in Table 3. Additional blood samples for PK analysis may be collected if clinically indicated (e.g., in case 
of SAE). For PK sample collection instruction/procedures, refer to the relevant manual. 
p. Blood samples for OGTT test will be collected under fasting conditions (10 hours) at the following timepoints: 0 minutes (just prior to ingesting glucose), 
30 minutes, 60 minutes, 90 minutes 120 minutes and 180 minutes. On Day 92/ET the insulin will be captured from the OGTT. 
q. At Screening, MRI-PDFF to be done within 35 days of baseline (Day 1). On Days 50 and 92, MRI-PDFF to be done within ± 2 days of the planned visit 
date. If out of the window, MRI-PDFF should still be performed as close to the target day as possible, but protocol deviation should be recorded. 
r. 
Samples for carboxy-terminal collagen crosslinks (CTX) and N-terminal propeptide of type 1 collagen (P1NP) will be obtained at the designated 
timepoints for storage and potential future analysis. The D42 sample will be obtained pre-dose. 
s. 
Samples for RNA as well as plasma and serum samples will be collected for potential future exploratory assessments, to increase understanding of 
BIO89-100 biological activity and to identify potential existing and/or emerging biomarkers. 
t. 
 
 
 
 
 
u. Immunogenicity sample on Day 92/ET will only be collected for subjects who early terminate from the study. 
v. Baseline samples of endogenous FGF21 will be analyzed;
 
 
w. The sites may take non-personally identifying photographs of potential injection site reactions (optional) 
x. Subjects in Cohorts 1 to 4 will be domiciled at the clinic from 1 day prior to dosing until 24 hours post the 1st dose and the 5th dose. Other study visits will 
be ambulatory. On Day 8, Day 15 and Day 22 (ambulatory dosing visits), subjects will remain at the site for observation for at least 2 hours post dosing. 


Protocol BIO89-100-002 
 
 
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Table 3 
Part 1 PK Sample Collection for Cohorts 1-4 – QW Dosing Interval 
Study Day 
Dosing 
Day 
Ambulatory 
visit 
Hours relative to 1st dose (Day 1) 
X 
 
 
 
 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
 
 
 
 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 
 


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Table 4 
Part 1 Schedule of Activities for Cohorts 5 and 6 
Cohorts 5-6 
Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit 
Study Day 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Medical 
history/demographics 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Percutaneous liver 
biopsy (optional)b 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Prior medications 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion and 
exclusion criteria 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Complete physical 
examc 
X 
 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
 
 
 
X  
X 
Symptom-directed 
physical examd 
 
 
X 
B 
P 
 
X 
B 
X 
 
B 
 
 
 
 
B 
 
B 
 
 
Body weighte 
X 
 
 
B 
 
 
X 
B 
X 
 
B 
 
 
 
B 
 
B 
 
 
Waist and hip 
measurements 
X 
 
 
B 
 
 
 
 
 
 
 
 
B 
 
 
 
 
X 
 
X 
X 
Fibroscanf 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
 
12-lead ECG 
(single)g 
X 
 
 
B 
X 
 
 
 
X 
B 
 
 
B 
X 
 
 
 
 
X 
X 
Urine drug screen 
and alcohol breath 
test 
X 
 
Xh 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Clinical laboratory 
testsi 
X 
X 
 
B 
X 
 
 
 
X 
B 
X 
 
B 
X 
 
 
 
X 
X 
X 
Cortisol (24 hour 
urine collection)j 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
X 
 
Urinalysis 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
X 
 
 
 
X 
X 
X 
HbA1C 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 


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Cohorts 5-6 
Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit 
Study Day 
Serologyk 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
TSH 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Pregnancy test in 
WOCBP onlyl 
X (S) 
 
X 
 
 
 
 
 
 
B 
 
 
B 
 
 
 
 
B 
 
B 
X 
X  
Vital signsm 
X 
 
X 
X 
P 
X 
X 
X 
X  
X 
X 
X 
X 
P 
X 
X 
X 
X 
X 
X 
X 
X 
Columbia-Suicide 
Severity Rating Scale 
(C-SSRS) 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
 
Randomization 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Study Intervention 
Administrationn 
 
 
 
X 
 
 
 
 
 
X 
 
 
X 
 
 
 
 
X 
 
X 
 
 
PK blood collectiono 
 
 
 
X 
X 
X 
X 
X 
X  
B 
X 
 
X 
X 
X 
X 
X 
 
X 
X 
 
PD and biomarker 
blood sampling: 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
IGF-1, total 
 
 
 
B 
 
 
 
 
X 
B 
 
 
B 
 
 
 
 
 
 
 
X 
 
Adiponectin  
 
 
 
B 
 
 
 
 
X 
 
 
 
B 
 
 
 
 
 
X 
 
X 
X 
CK-18, ELF panel 
and Pro-C3 
 
 
 
B 
 
 
 
 
 
 
 
 
B 
 
 
 
 
 
X 
 
X 
Free fatty acid 
 
 
 
B 
 
 
 
 
X 
 
 
 
B 
 
 
 
 
 
X 
 
X 
X 
Oral glucose 
tolerance test 
(OGTT)p  
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
Insulin (not part of 
OGTT) 
 
 
 
B 
 
 
 
 
X 
 
 
 
B 
 
 
 
 
 
X 
 
 
X 
HOMA-IR 
calculation 
 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 


Protocol BIO89-100-002 
 
 
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Cohorts 5-6 
Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit 
Study Day 
MRI-PDFF; visceral 
fat; and 
subcutaneous fat q 
X  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
 
Plasma sample for 
potential future bone 
biomarkers analysisr 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
Pharmacogenomic 
(DNA) blood 
sampling 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Exploratory 
biomarker analysis s 
 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
X 
X 
Immunogenicityt 
 
 
 
B 
 
 
 
 
 
B 
 
 
B 
 
 
 
 
 
X 
 
Xu 
X 
Endogenous FGF21v 
 
 
 
B 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Adverse event 
monitoringw 
X 
X 
X 
←-===========================================→ 
X 
X 
Concomitant 
medication  
X 
X 
X 
←-===========================================→ 
X 
X 
Domiciledx 
 
 
X 
X 
X 
 
 
 
 
X 
X 
X 
 
 
 
 
 
 
 
 
Abbreviations: B = predose; D = Day; CK = cytokeratin; DNA = deoxyribonucleic acid; ECG=electrocardiogram; ELF = enhanced liver fibrosis; EOS = end of study; 
ET= early termination; FGF21 = fibroblast growth factor 21; FU = Follow-Up; HbA1c=hemoglobin A1c; IGF-1 = insulin-like growth factor; MRI-PDFF = magnetic 
resonance imaging based proton density fat fraction; P = Pre-discharge; PK=pharmacokinetic; Pro-C3 = N‐terminal propeptide of type III collagen; S = serum; TSH = 
thyroid stimulating hormone; WOCBP=women of child-bearing potential. 
Table 4 footnotes 
All outpatient visits will have a study window of ±1 day 
* Between Visits 17 to 19, on Days 64 and 78 subjects will be contacted by phone to inquire about AEs and concomitant medications 
a. 
For any subject who withdraws before completion of the study, an Early Termination (ET) visit will be conducted, if possible; with the same assessments 
as the Day 92 visit.  
b. Optional liver biopsy may be performed  for subjects who do not have a medical contraindication to undergoing a liver biopsy, and did not have a liver 
biopsy in the 24 months prior to screening.|For subjects who underwent a previous liver biopsy >24 months prior to screening, that would not have 


Protocol BIO89-100-002 
 
 
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qualified for the study based on results of the initial biopsy, there should be a sound clinical basis to expect different findings in a repeat biopsy based on 
medical judgment. Any biopsy will need to be approved in advance by Sponsor and only after the subject has met all other inclusion and exclusion 
criteria. 
c. 
Complete physical exam done at screening to include recording height, weight, and calculating BMI. 
d. Symptom-directed physical exam will be done pre-dose and prior to discharge from the Phase 1 unit on domiciled dosing visits (1st and 3rd dose), and 
pre-dose on ambulatory dosing visits. Additional physical examinations will also be performed if clinically indicated. 
e. 
On domiciled and ambulatory visits, body weight will be measured pre-dose. 
f. 
Fibroscan should be performed during screening, prior to magnetic resonance imaging (MRI) for all subjects.  
g. 12-lead safety ECGs will be recorded as single bedside measurements. On domiciled dosing visits (1st and 3rd dose), ECG will be measured pre-dose and 
24 hours post-dose; on ambulatory dosing visits, ECG will be measured pre-dose. Additional ECG may be conducted if clinically indicated. 
h. On Day -1, urine drug screen can be done at a local laboratory, however, a sample should also be collected for central laboratory evaluation  
i. 
Clinical laboratory tests will include biochemistry, hematology, and coagulation, and FSH for determination of post-menopausal status ; on domiciled 
dosing visits (1st and 3rd dose), clinical laboratory will be collected pre-dose and 24 hours post dose; on ambulatory visits, clinical laboratory will be 
collected pre-dose. For all subjects, alanine transaminase (ALT) and aspartate transaminase (AST) will be collected twice during the Screening period (at 
least 2 weeks apart), with the 2nd assessment to be collected at Day -7 to Day -3. A 3rd assessment, if required, will be collected via unscheduled visit 
(refer to Exclusion criterion 3). 
j. 
Ambulatory 24 hour urine collection for cortisol to be done within 14 days from baseline, on Day 50 and Day 92/ET. It is recommended that subjects 
collect urine for the 24 hours prior to D-7 to-3 visit and bring the container with them to this visit. Alternatively, urine can be collected for the 24 hours 
prior to the randomization visit (D-1) and brought to the site on D-1 (the result is not required for eligibility confirmation).  For Day 50 and D92/ET – 
subject will start collection 24 hours before coming into the clinic and bring the sample to the visit. 
k. Serology tests will include Hepatitis B surface antigen, Hepatitis C Virus, and Human Immunodeficiency Virus (HIV) 1 and 2 antibodies 
l. 
Serum urine pregnancy test will be conducted at screening; at all other timepoints urine pregnancy test will be done to guide clinical decisions to dose on 
dosing days. Prior to the 1st dosing, the baseline urine pregnancy test will be performed on D-1 to allow for randomization on that day.  If urine test is 
positive, a confirmatory serum pregnancy test will be conducted. 
m. Vital signs include supine blood pressure, pulse, body temperature, and respiratory rate; vital signs will be measured pre-dose (prior to scheduled blood 
draws and study intervention administration); 1, 12 and 24 hours post-dose and prior to discharge on domiciled dosing visits (1st and 3rd doses); on dosing 
ambulatory visits, vital signs will be measured pre-dose (prior to scheduled blood draws and study intervention administration) and prior to discharge; on 
non-dosing visits, vital signs will be measured prior to scheduled blood draws. Starting from randomization, blood pressure and pulse will be measured in 
duplicate, the first measurement will be taken up to 15 minutes before the indicated timepoint. Additional vital signs measurement may be done if 
clinically indicated. Subjects must be in a supine or semi-erect/seated position and resting for at least 5 minutes prior to measurements. 
n. Study intervention will be administered SC to the abdomen region by qualified study personnel.  
o. PK blood samples will be collected as shown in Table 5. Additional blood samples for PK analysis may be collected if clinically indicated (e.g., in case 
of SAE). For PK sample collection instruction/procedures, refer to the relevant manual. 
p. Blood samples for OGTT test will be collected under fasting conditions (10 hours) at the following timepoints: 0 minutes (just prior to ingesting glucose), 
30 minutes, 60 minutes, 90 minutes 120 minutes and 180 minutes. . On Day 92/ET the insulin will be captured from the OGTT. 
q. At Screening, MRI-PDFF to be done within 35 days of baseline (Day 1). On Days 50 and 92, MRI-PDFF to be done within ± 2 days of the planned visit 
date. If out of the window, MRI-PDFF should still be performed as close to the target day as possible, but protocol deviation should be recorded. 
r. 
Samples for carboxy-terminal collagen crosslinks (CTX) and N-terminal propeptide of type 1 collagen (P1NP) will be obtained at the designated 
timepoints for storage and potential future analysis. The D42 sample will be obtained pre-dose 


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s. 
Samples for RNA as well as plasma and serum samples will be collected for potential future exploratory assessments, to increase understanding of 
BIO89-100 biological activity and to identify potential existing and/or emerging biomarkers. 
 
 
 
 
 
u. Immunogenicity sample on Day 92/ET will only be collected for subjects who early terminate from the study.  
v. Baseline samples of endogenous FGF21 will be analyzed; 
 
  
w. The sites may take non-personally identifying photographs of potential injection site reactions (optional) 
x. Subjects in Cohort 5 and 6 will be domiciled at the clinic from 1 day prior to dosing until 24 hours post the 1st dose and 24 hours post the 3rd dose; other 
study visits will be ambulatory. On the Day 15 ambulatory dosing visit, subjects will remain at the study site for observation for at least 2 hours post 
dosing. 
 
 


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Table 5 
Part 1 PK Sample Collection for Cohorts 5 and 6 – Q2W Dosing Interval 
Study Day 
Dosing Day 
Ambulatory visit 
Hours relative to 1st dose (Day 1) 
X 
 
 
 
 
X 
 
X 
 
X 
 
X 
X 
X 
 
X 
X 
 
 
 
 
X 
 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
X 


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1.2.2. 
Part 2 
Table 6 
Schedule of Activities for Part 2 (Cohort 7) 
Cohort 7 
Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit 
 Study Day 
(mandatory in clinic visits) 
Informed consent 
X 
 
 
 
 
Medical history/demographics 
X 
 
 
 
 
Percutaneous liver biopsyb 
X 
 
 
X 
 
Histology machine read (PathAI) 
X 
 
 
X 
 
Prior medications 
X 
 
 
 
 
Inclusion and exclusion criteria 
X 
 
B 
 
 
Complete physical examc 
X 
 
 
X  
X 
Symptom-directed physical examd 
 
 
B 
 
 
Body weight 
X 
 
B 
X 
X 
Waist and hip circumference 
X 
 
B 
X 
 
Lifestyle counseling 
X 
X 
X 
 
 
Fibroscane 
X 
 
 
X 
 
12-lead ECG (single)f 
X 
 
B 
X 
X 
Urine drug screen and alcohol breath 
test 
X 
 
Bg 
 
 
Clinical laboratory testsh 
X 
X 
B 
X 
X 
Urinalysis 
X 
 
B 
X 
X 
HbA1C 
X 
 
B 
X 
X 
Serologyi 
X 
 
 
 
 
TSH 
X 
 
 
 
 
Pregnancy test in WOCBP onlyj 
X(S) 
 
B 
X 
X 
Vital signsk 
X 
 
B 
X 
X 


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Cohort 7 
Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit
 Study Day
(mandatory in clinic visits)
Columbia-Suicide Severity Rating 
Scale (C-SSRS) 
X 
 
 
 
 
 
Study intervention in-clinic 
(mandatory) administrationl 
 
 
X 
X 
 
 
Home or in-clinic study intervention 
administrationm 
 
 
 
 
 
PK blood collectionn 
 
 
B 
X 
 
Pro-C3 
 
 
B 
X 
 
Adiponectin 
 
 
B 
X 
 
MRI-PDFFo  
X 
 
 
X 
 
Bone biomarkers analysisp 
 
 
B 
X 
X 
Immunogenicity q 
 
 
B 
X 
X 
Endogenous FGF21r 
 
 
B 
 
 
 
Adverse event monitorings 
X 
X 
X 
 ←===============X ==============→ 
X 
X 
Concomitant medication 
X 
X 
X 
←================X ===============→ 
X 
X 
Abbreviations: B = predose; D = Day; AI = artificial intelligence; ECG=electrocardiogram; EOS = end of study; ET= early termination; FGF21 = fibroblast growth 
factor 21; FU = Follow-Up; HbA1c=hemoglobin A1c; MRI-PDFF = magnetic resonance imaging based proton density fat fraction; PK=pharmacokinetic; Pro-C3 = 
N-terminal propeptide of type III collagen; S = serum; TSH = thyroid stimulating hormone; WOCBP=women of child-bearing potential. 
Table 6 Footnotes 
All ambulatory clinic visits will have a study window of ±1 day.  Window for MRI-PDFF will be ±7 days. The first liver biopsy will be done during the screening 
period. The second liver biopsy will be performed within 14 days after the last dose of study intervention. Liver biopsies (first and second) will be done after MRI-
PDFF.  
a. 
For any subject who withdraws before completion of the study, an Early Termination (ET) visit will be conducted, if possible; with the same assessments 
as the Day 141 visit.  
b. A liver biopsy with NASH that meets study inclusion criteria, performed within 24 weeks from baseline visit with the sample deemed interpretable by the 
central reader is acceptable instead of the baseline liver biopsy. A second liver biopsy will be done within 14 days of the last dose of study intervention. 


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Liver biopsies (first and second) will be done after MRI-PDFF. If out of a window, a liver biopsy should still be performed as close to the target date as 
possible, but a protocol deviation should be recorded.  
c. 
Complete physical exam done at screening to include recording height, weight, and calculating BMI. 
d. Physical examinations will be performed if clinically indicated. 
e. 
Fibroscan should be performed during screening, prior to magnetic resonance imaging (MRI) for all subjects. Fibroscan will be repeated on D141/ET. 
f. 
12-lead safety ECGs will be recorded as single bedside measurements. Additional ECG may be conducted if clinically indicated. 
g. On Day 1, urine drug screen can be done at a local laboratory. 
h. Clinical laboratory tests (performed under fasting conditions, ≥10 hours) will include biochemistry, hematology, and coagulation, and FSH for 
determination of post-menopausal status. For all subjects, alanine transaminase (ALT) and aspartate transaminase (AST) will be collected twice during 
the Screening period (at least 2 weeks apart), with the 2nd assessment to be collected at Day -7 to Day -3. A 3rd assessment, if required, will be collected 
via unscheduled visit (refer to Exclusion criterion 3). 
i. 
Serology tests will include Hepatitis B surface antigen, Hepatitis C Virus, and Human Immunodeficiency Virus (HIV) 1 and 2 antibodies. 
j. 
Serum urine pregnancy test will be conducted at screening; at all other timepoints urine pregnancy test will be done to guide clinical decisions to dose on 
dosing days. On dosing days in the clinic, a urine pregnancy test will be obtained locally; in addition, a urine sample will be sent to the central lab. If 
urine test is positive, a confirmatory serum pregnancy test will be conducted. 
k. Vital signs include blood pressure, pulse, body temperature, and respiratory rate; vital signs will be measured pre-dose (prior to scheduled blood draws 
and study intervention administration. Blood pressure and pulse will be measured in duplicate, the first measurement will be taken up to 15 minutes 
before the indicated timepoint. Additional vital signs measurement may be done if clinically indicated. Subjects must be in a supine or semi-erect/seated 
position and resting for at least 5 minutes prior to measurements. 
l. 
Study intervention will be administered SC to the abdomen region by qualified study personnel or by the subject (under supervision) at ambulatory clinic 
visit and will be administered at home by the subject at the rest of the planned dosing days. Subjects will be observed for 15 minutes after each dose. 
m. At the designated timepoints, study intervention will be administered at the subject’s home. If home administration is not feasible, not desired by the 
subject, or deemed inappropriate option by the investigator for any reason, in-clinic study intervention administration may take place at some or all of 
these timepoints.   
n. PK blood samples will be collected at the specified timepoints. Additional blood samples for PK analysis may be collected if clinically indicated (e.g., in 
case of SAE). For PK sample collection instruction/procedures, refer to the relevant manual. 
o. At Screening, MRI-PDFF to be done within 35 days of baseline (Day 1). On Days 85 and 141, MRI-PDFF to be done within ± 7 days of the planned visit 
date. If out of the window, MRI-PDFF should still be performed as close to the target day as possible, but protocol deviation should be recorded. 
p. Samples for carboxy-terminal collagen crosslinks (CTX), N-terminal propeptide of type 1 collagen (P1NP) and osteocalcin will be obtained at the 
designated timepoints.  
q. 
 
 
 
 
 


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r. 
Baseline samples of endogenous FGF21 will be obtained in all subjects;
 
 
s. 
The sites may take non-personally identifying photographs of potential injection site reactions (optional). 


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2. 
INTRODUCTION 
2.1. 
Study Rationale 
BIO89-100, developed by 89bio, is a glycoPEGylated analog of FGF21, a metabolic hormone 
secreted by the liver, adipose tissue, skeletal muscle, and the pancreas. FGF21 is regulated by 
nutritional status, affects energy expenditure, and glucose and lipid metabolism (Lee, 2014) 
(Arner, 2008) (Park, 2016), and therefore may play a role in metabolic disease, e.g., NAFLD, a 
common chronic liver disease affecting 30% of adult population worldwide (Chalasani, 2012). 
Several nonclinical and clinical studies have shown that administration of similar FGF21 analogs 
had beneficial effects on serum lipid and insulin-resistance as well as on liver fat (Zhang 2014) 
(Gaich, 2013) (Sanyal, 2019). There are currently no approved pharmaceutical treatments for 
NASH (Friedman, 2018). 
This Phase 1b/2a study has 2 parts. Part 1 is designed to assess the safety and tolerability of 
repeated SC escalating doses of BIO89-100 in subjects with a diagnosis of NASH, or who have 
NAFLD with a high risk of NASH. Part 2 is open-label, designed to assess the effect of repeated 
SC doses of one dose level of BIO89-100 (27 mg QW), administered for 20 weeks, on liver 
histology in subjects with biopsy-proven NASH with NAFLD Activity Score (NAS) ≥4, fibrosis 
stage F1 with high risk, F2 or F3.  
The study includes safety, PK, PD, and immunogenicity characterizations to allow a preliminary 
assessment of a safe and efficacious dose range to be studied in further clinical studies. 
2.2. 
Background 
2.2.1. 
Non-alcoholic Steatohepatitis 
NASH is a chronic liver disease, characterized histologically by hepatic steatosis in ≥5% of 
hepatocytes, injury (ballooning)(Chalasani, 2012). It is part of the spectrum of NAFLD that 
includes NASH, and cirrhosis resultant from fatty liver (Estes, 2018). In North America, the 
prevalence of NAFLD is estimated at ~24%. NAFLD patients tend to be obese, with insulin 
resistance and/or type 2 diabetes mellitus (T2DM), dyslipidemia, hypertriglyceridemia, and 
hypertension, and NAFLD is increasingly recognized as the liver disease component of the 
metabolic syndrome (MetS) (Chalasani, 2018).  
The pathophysiologic mechanisms that lead to NAFLD are thought to be similar to those in the 
development of T2DM, as insulin resistance is a hallmark in both disease states. Insulin 
resistance in adipocytes leads to dysfunction in the normal regulation of lipolysis, contributing to 
elevated circulating free fatty acids, accumulation of triglycerides in the liver (Saponaro, 2015) 
and lipotoxicity, manifested as inflammation and cellular injury (Byrne, 2015), and ultimately – 
fibrosis. 
NAFLD is usually asymptomatic, unless progression to cirrhosis has occurred. It is often 
diagnosed by demonstration of hepatic steatosis on liver imaging (e.g., ultrasound or magnetic 
resonance imaging (MRI)) in subjects, commonly with features of the metabolic syndrome, in 
whom no alternative etiology for liver fat accumulation can be identified (e.g., alcoholic liver 
disease, medications). Validated noninvasive tests for diagnosis of steatohepatitis are not 


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currently available, and a liver biopsy is still needed to diagnose the inflammation and cellular 
ballooning features of NASH (Torres, 2012). 
There is a high unmet need in NASH (Friedman, 2018). The disease progresses to fibrosis and 
cirrhosis in approximately 20% of patients (Vernon, 2011; Khan, 2015), and 45% of patients 
with cirrhosis will progress to decompensated cirrhosis within 10 years (Rinella, 2015). Eight 
percent of patients with advanced fibrosis will develop hepatocellular carcinoma (HCC) within 
5 years (Hashimoto, 2009), and there is increasing evidence that non-cirrhotic subjects with 
NAFLD may also be at an increased risk of HCC (Mittal, 2016). Pre-cirrhotic NASH has a 
relative risk of death of 1.7 over 15 years compared to the general population, and approximately 
20% of NAFLD patients die of liver-related causes (Soderberg, 2010). A variety of therapeutic 
agents are being developed for NASH, targeting metabolic pathways, inflammatory pathways or 
fibrosis. Positive results from the REGENERATE study, the first and largest successful pivotal 
Phase 3 study in patients with liver fibrosis due to NASH have been reported (Younossi, 2019). 
However, as of this time, there are no approved therapies for treatment of patients with NASH. 
2.2.2. 
FGF21  
FGF21 is a metabolic hormone, regulated by nutritional status, that affects energy expenditure 
and glucose and lipid metabolism. It is secreted mainly by the liver, but also by white adipose 
tissue, skeletal muscle, and the pancreas. The activation of fibroblast growth factor receptors 
(FGFRs) by FGF21 requires the transmembrane protein cofactor β-Klotho, expressed 
predominantly in metabolic organs, including the liver, white adipose tissue, and the pancreas, 
conferring organ specificity to FGF21 (Ogawa, 2007; Li, 2015). 
In patients with insulin resistance and NASH, circulating and tissue levels of FGF21 are 
increased and correlate with disease severity, indicating the presence of FGF21 resistance, which 
can be overcome by administration of pharmacological doses of FGF21. On this basis, 
administration of exogenous FGF21 has been explored as a method to treat obesity-associated 
insulin-resistance disorders, including NASH. 
Native FGF21 has a short life (~2 hours), limiting the potential to use it as a therapeutic agent. 
Consequently, various methods have been developed to extend the half-life of FGF21, including 
PEGylation. PEGylated FGF21 was shown to provide comparable efficacy to wild-type FGF21 
despite lower dosing frequency and total cumulative dose, a likely result of greatly increased 
in vivo half-life (Mu, 2012). In humans, the effects of pegbelfermin (BMS-986036; a PEGylated 
analog of FGF21, which differs from BIO89-100 in 
 
) on biopsy confirmed NASH patients (both genders; fibrosis stage 
F1-F3) with hepatic fat fraction ≥10% (per MRI-PDFF) were recently assessed in a 16-week 
randomized placebo-controlled study (n=74) (Sanyal, 2019). The study demonstrated a 
statistically significant reduction in liver fat versus placebo at week 16 (primary endpoint; 
measured by MRI-PDFF), with up to 6.8% (p=0.0004) and 5.2% (p=0.008) absolute reduction 
for a 10 mg daily dose and 20 mg weekly dose, respectively. The study also showed 
improvements in Pro-C3, serum biomarker of fibrosis, magnetic resonance elastography 
(measure of liver stiffness), adiponectin, and liver enzymes (ALT, and AST). Improvements in 
triglycerides, LDL-c, and HDL-c were also noted primarily with the 10 mg daily dose. Finally, 
safety was considered favorable, with no deaths, treatment-related SAEs, or discontinuations due 
to AEs reported. The most frequently reported AEs were diarrhea, nausea and frequent bowel 
movement, most of these were mild, and none were considered as severe in intensity. 


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2.2.3. 
BIO89-100 
BIO89-100 is a 
PEGylated analog of FGF21 that has an
 
 It 
was evaluated in nonclinical pharmacology, PK, and toxicology studies and in a recently 
completed, first-in-human, single ascending dose study (TV47948-SAD-10122). 
In nonclinical pharmacology studies, BIO89-100 was found to be effective in two NASH models 
in mice and in spontaneously diabetic monkeys, with liver-related and metabolic benefits, 
including improved transaminase levels and liver histology, decreased body weight, improved 
glycemic parameters and lipid profile, and increased adiponectin levels. Nonclinical safety 
studies with BIO89-100 have shown good tolerability to treatment at doses that were time-folds 
higher than the effective doses in animal pharmacological models and over systemic human 
exposure to BIO89-100 following single administration of clinical doses (TV47948-SAD-10122 
study). Data from repeated nonclinical toxicology GLP studies in mice suggest that at high 
doses, BIO89-100 may have an adverse effect on bone formation and linear growth, and on the 
female reproductive system (refer to Section 2.3.1). The determined no observed adverse effect 
level (NOAEL) in males and female mice is much higher than the suggested clinical doses in 
BIO89-100-002. 
TV47948-SAD-10122, a Phase 1, first-in-human, randomized, double-blind, placebo-controlled 
study to evaluate the safety, tolerability, and PK properties of BIO89-100 following SC 
administration of single ascending doses (SAD), has recently been completed. The study 
randomized and treated 58 subjects, 43 of whom were dosed with BIO89-100 at 7 dose levels 
(0.45 mg; 1.2 mg; 3 mg; 9.1 mg; 18.2 mg; 39 mg; and 78 mg). No significant safety issues have 
been identified. 
 
 
 
 
 
A detailed description of the chemistry, pharmacology, efficacy, and safety of BIO89-100 is 
provided in the Investigator’s Brochure (IB). 
2.3. 
Benefit / Risk Assessment 
Information about the expected benefits and potential risks (adverse event) of BIO89-100 can be 
found in the IB. 
2.3.1. 
Risk Assessment 
BIO89-100 has not been administered previously to patients with NASH or NAFLD who are at 
high risk for NASH. Mild injection site reactions and sporadic GI effects have been reported in a 
recently completed SAD study (Study TV47948-SAD-10122) with BIO89-100. Potential risks 
based on nonclinical data with BIO89-100 and published class effects include immunogenicity, 
bone effects and female reproductive effects. It is important to note that none of these potential 
risks have been reported as significant safety concerns in the recently published Phase 2 proof-
of-concept study in which subjects with biopsy-proven NASH were treated for 16 weeks with 


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pegbelfermin, a PEGylated FGF21 analogue. Additional information regarding risks to subjects 
with BIO89-100 may be found in the IB. 
2.3.1.1. 
Potential Risk: Immunogenicity 
As with any biologic drug, exposure to BIO89-100 may potentially provoke an immune 
response, with formation of ADAs, that may lead to loss of efficacy or adverse effects (e.g., 
hypersensitivity, loss of endogenous FGF21 activity). After single administrations to healthy 
subjects in study TV47948-SAD-10122, 
 of BIO89-100-treated subjects had positive 
ADA, all with 
 ADAs to BIO89-100 have 
 
, without a discernible effect on PK or PD 
parameters that were evaluated in these studies. No severe hypersensitivity event has been 
reported in Cohorts 1-6 (completed). Of note, formation of ADAs in nonclinical species is 
expected, and is generally not considered predictive for immunogenicity in humans. 
In study BIO89-100-102, the following mitigation measures will be implemented to address the 
potential risk of severe hypersensitivity reactions, including anaphylaxis: 
a. Subjects with any prior exposure to an FGF21 analogue, if known, will be excluded 
from participating in the study.  
b. In Part 1, subjects will be under observation and intensive monitoring (including 
multiple vital signs measurements) for 48 hours in 2 domiciled visits (1st and 5th dose 
for Cohorts 1-4, 1st and 3rd dose for Cohorts 5 and 6), and for 2 hours post dose during 
ambulatory dosing visits in the first month. In Part 2, subjects will be observed for at 
least 15 minutes after each dose of study intervention. 
c. All study sites will have appropriate measures (personnel and equipment) to diagnose 
and address severe hypersensitivity reactions, including proximity to a hospital to 
which a subject may be evacuated, if needed. Details on identification and 
management of severe hypersensitivity reactions after home administration will be 
provided in the Home Administration Manual.    
d. Severe hypersensitivity reactions (including anaphylaxis) will be an event to monitor.  
e. Stopping rules include serious hypersensitivity reactions to BIO89-100.  
f. Hypersensitivity-related AEs will be captured as part of the general safety assessment 
in all subjects. 
g. Immunogenicity testing will be done frequently, at baseline and multiple timepoints 
during study treatment. In addition, an immunogenicity sample will be obtained in the 
event of a significant hypersensitivity reaction. Subjects who test positive for 
neutralizing antibodies to BIO89-100 at the EOS/ET visit will be asked to return for 
additional follow-up testing. This testing should occur approximately every 
 
 
. More frequent testing (e.g., every 
month) or testing for a longer period of time may be requested in the event of safety-
related concerns. Follow-up testing will not be required where it is established that 
the subject did not receive BIO89-100 (Part 1). All follow-up results, both positive 
and negative, will be communicated to the sites. A blood sample for ADA assessment 
will also be collected upon observation of any severe hypersensitivity reaction (e.g., 
anaphylaxis). Subjects who test positive for binding, non-neutralizing antibodies and 


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have clinical sequelae that are considered potentially related to an anti-BIO89-100 
antibody response may also be asked to return for additional follow-up testing. 
2.3.1.2. 
Potential Risk: Bone Effects 
The literature suggests that FGF21 may be involved in bone turnover (Wei, 2012; Owen, 2013). 
In nonclinical studies with BIO89-100 in mice, non-adverse reductions in bone formation and 
resorption markers were observed in young, healthy mice, as well as decreases in tibia size and 
bone density. The relevance of these findings to humans is not known. Interestingly, in a study in 
40 healthy volunteers, fasting plasma FGF21 levels correlated positively with total bone mineral 
density (BMD) (R2=0.69, p=0.003) and spine BMD (R2=0.76, p=0.001) in women, a correlation 
that remained significant after adjusting for age, ethnicity, and body composition; there was no 
association between FGF21 concentrations and body composition in men (Lee, 2013). 
Bone-related assessments have been conducted in studies of 2 long-acting FGF analogues in 
humans: PF-05231023, a long-acting synthetic FGF21 analogue generated by covalent linking of 
2 recombinant human FGF21 molecules to the Fab portion of a proprietary scaffold antibody, 
and pegbelfermin, a PEGylated FGF21 analogue. Modest changes in bone turnover markers were 
observed with PF-05231023 over 4 weeks of treatment (Kim, 2017), especially in the context of 
weight loss, although the clinical significance of these findings is not established. With 
pegbelfermin no bone findings were observed relative to placebo treatment over 16 weeks of 
treatment. 
Bone-related AEs were not reported in a single ascending dose study with BIO89-100 (study 
TV47948-SAD-10122) however, this short-term study may not be informative for this type of 
risk. The determined NOAEL of BIO89-100 in males and female mice (up to 13 weeks) and 
monkeys (up to 28 days) is much higher than the suggested clinical doses in BIO89-100-002. It 
is notable that in addition to the presence of an adequate safety margin relative to the highest 
dose to be administered in study BIO89-100-102, the risk of a clinically significant impact on 
bone is considered to be low in the setting of a 12-week study, as significant changes to bone 
typically require longer exposures. Nevertheless, as a precautionary measure, in study 
BIO89-100-102 subjects younger than 21 year of age and subjects with a history of bone trauma, 
bone fracture, or bone surgery within 2 months of screening will be excluded. In Part 2, 
exclusion of subjects with bone disorders, including as osteoporosis, osteomalacia, and known, 
untreated severe vitamin D deficiency (serum 25-hydroxy-vitamin D ≤5 ng/mL) has been added 
as an additional precautionary measure, given the longer exposure.  
Bone-related AEs will be captured as part of the general safety assessment in all subjects. Plasma 
samples will be collected for potential future analysis of bone turnover biomarkers, if deemed 
necessary. In Part 2, samples of bone turnover biomarkers will be analyzed. 
2.3.1.3. 
Potential Risk: Female Reproductive Effects 
In nonclinical studies in mice exposed to high doses of BIO89-100,
 
. These are known 
effects of FGF21 in mice that have previously been described in the literature (Wei, 2012;, 
Owen, 2013). Importantly, the established NOAEL of 15 mg/kg in female mice provides an 
adequate safety margin (16.2- and 52.4-fold margin for AUC168, ss and Cmax for the 27 mg QW 
clinical dose) for the female reproductive findings that were observed in nonclinical studies with 


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BIO89-100. Consequently, women of childbearing potential (WOCBP) will be allowed to enroll 
in study BIO89-100-102. It is worth noting that WOCBP were enrolled in the recent Phase 2a 
study with pegbelfermin (BMS-986036); no reproduction-related AEs were reported (Sanyal, 
2019). Female reproductive AEs were not reported in a single ascending dose study with BIO89-
100 (study TV47948-SAD-10122). Specific assessments of the female reproductive system were 
not performed in this short-term study, which enrolled only females who were not of 
childbearing potential, and the study is not considered to be informative for this type of risk. 
WOCBP and their partners will be required to use highly effective contraception, as per current 
standards for investigational products with an unknown potential for embryotoxicity. Female 
reproductive system-related AEs will be captured as part of the general safety assessment in all 
female subjects. 
2.3.1.4. 
Potential Risk: Injection Site Reactions 
As with other protein drugs that are administered subcutaneously, there is a potential for 
injection site reactions (erythema, induration, ecchymosis, tenderness, warmth, swelling, and - at 
times – more severe reactions, such as lipodystrophy, ulceration, or necrosis). 
Injection site reactions following subcutaneous BIO89-100 injections in nonclinical species were 
non- adverse and reversible after a treatment-free period. Some mild (CTCAE Grade 1) injection 
site reactions were reported in a single ascending dose study with BIO89-100 (study TV47948-
SAD-10122) that completely resolved and did not require treatment; few injection site reactions 
were also reported in the placebo group.
 
 
 
2.3.1.5. 
Potential Risk: Gastrointestinal Effects 
Gastrointestinal (GI) effects have been reported in clinical studies with other FGF21 analogues. 
In a recent Phase 2a study with pegbelfermin (BMS-986036), in which NASH patients were 
administered 10 mg daily or 20 mg weekly regimens of pegbelfermin (N=49) or placebo (N=26), 
the most frequently reported AEs were diarrhoea and nausea, which were more common in 
pegbelfermin-treated than in placebo-treated patients. There were 28 GI-related AEs, including 
diarrhoea, nausea, frequent bowel movements, upper abdominal pain, and vomiting, 11 of which 
were considered to be treatment-related. The frequency of GI AEs was higher in patients treated 
with pegbelfermin than in patients who received placebo but there was no clear association 
between frequency of AEs and pegbelfermin dose. All GI events were considered mild or 
moderate and only 1/11 treatment-related GI AEs required treatment for resolution (probiotics). 
No GI-related effects were observed in nonclinical studies with BIO89-100 in either mice or 
monkeys.  
A few mild, sporadic GI effects (nausea, dyspepsia, abdominal pain, constipation, diarrhea, 
flatulence, frequent bowel movements), most of which were considered not related to study drug 
and all of which resolved without treatment, were reported in a single ascending dose study with 
BIO89-100 (study TV47948-SAD-10122). 


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2.3.1.6. 
Potential Risk of Liver Biopsy 
Following liver biopsy, the overall rate of serious complications has been reported to be 
approximately 1% (Seeff, 2010; Boyum, 2016) and the overall mortality risk has been estimated 
to be 0.2% (West, 2010). Most complications (83% to 96%) occur within 24 hours of the 
procedure; 60% occur within 2 hours (Piccinino, 1986; Boyum, 2016). Approximately 2% to 3% 
percent of patients require hospital admission for management of complications; pain or 
hypotension are the predominant causes (Garcia-Tsao, 1993; Janes, 1993). 
Pain is the most common complication following percutaneous liver biopsy with approximately 
25% of patients experiencing pain in the right upper quadrant or right shoulder (Rockey, 2009). 
The pain usually resolves completely within a few hours.  
The most common serious complication of liver biopsy is intraperitoneal hemorrhage. Severe 
bleeding that results in hemodynamic compromise or requires intervention has been reported 
following 0.01% to 0.5% of liver biopsies (Boyum, 2016). Bleeding usually becomes clinically 
apparent within 3 to 4 hours after the biopsy. Hypotension and/or tachycardia following a biopsy, 
particularly when associated with abdominal pain, are usually related to hemorrhage. 
2.3.2. 
Benefit Assessment 
There are currently no approved products for NASH in the US. As discussed in previous 
sections, the FGF21 receptor is a promising mode of action for the treatment of NASH, and 
several FGF21 analogues are being evaluated for this indication. Recently, encouraging clinical 
data has been reported for a Phase 2 proof-of-concept study with pegbelfermin, a PEGylated 
FGF21 analogue, that was administered to patients with biopsy-proven NASH for 16 weeks 
(Sanyal, 2019). In this study, a significant reduction of hepatic fat (as assessed by MRI-PDFF) 
was evident, as well as improvement in fibrosis-related and metabolic parameters. While no 
clinical data for BIO89-100 are yet available, nonclinical data in mice and monkeys are 
promising, with significant pharmacological effects on liver-related and metabolic parameters in 
disease animal models. Taken together, it is estimated that 12 weeks of treatment with 
BIO89-100 may have a positive impact on liver-related (e.g., reduction of hepatic fat) and 
metabolic parameters in patients with NASH or with NAFLD at a high risk for NASH (Part 1), 
and 20 weeks of treatment with BIO89-100 may have a positive impact on liver histology in 
patients with biopsy-proven NASH and fibrosis (Part 2). These patients may thus derive benefit 
from this treatment. 
2.3.3. 
Update: Preliminary Data from Part 1 
At the time of finalization of this amendment, preliminary top-line data from Part 1 of this study 
(Cohorts 1-6) have become available. Analyses of the full dataset are ongoing.  
At Week 13, all BIO89-100 dose groups (Cohorts 1-6) showed significant absolute and relative 
reductions in MRI-PDFF. Up to 88% of BIO89-100 subjects achieved ≥30% MRI-PDFF 
reduction vs. baseline (p<0.001), a threshold of MRI-PDFF relative reduction that has been 
shown to be associated with histological benefit (Loomba, 2015; Loomba, 2017; Harrison, 
2019). Significant decreases in ALT vs. placebo were observed with BIO89-100, which were 
maximal with 27 mg QW (30 U/L decrease from baseline, p<0.001) and also prominent in the 
subgroup of subjects (n=17) with baseline ALT >45 U/L (35 U/L decrease from baseline, 
p<0.05). Reductions in Pro-C3, a fibrosis marker, were also noted. Metabolic benefits of 


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BIO89-100 included a favorable effect on lipids, with significant reductions in triglycerides (up 
to 28% in overall population, up to 49% in the subgroup [n=15] with baseline TG ≥200 mg/mL); 
non-HDL cholesterol and LDL-C (up to 15% and 16%, respectively), and increased adiponectin 
(up to +61%). 
BIO89-100 had a favorable tolerability profile. There were no deaths or related SAEs. One 
BIO89-100-treated subject discontinued due to a related AE (localized skin rash). Mild increased 
appetite (15.9% in pooled BIO89-100-treated subjects) was the most common AE. The 
frequency of gastrointestinal AEs compared favorably to placebo; diarrhea (BIO89-100 12.7%; 
placebo 22.2%) and nausea (BIO89-100 7.9%; placebo 16.7%) were the only GI AEs in ≥5% of 
BIO89-100-treated subjects. Injection site reactions, all mild, were reported in 6.3% of pooled 
BIO89-100-treated subjects. No hypersensitivity reactions were reported.  
2.3.4. 
Overall Benefit Risk Assessment 
As discussed, patients with NASH or with NAFLD at high risk for NASH (Part 1) or biopsy-
proven NASH (NAS ≥4 and fibrosis stage F1 with high risk, F2 or F3; Part 2) may derive benefit 
from treatment with BIO89-100. The preliminary data from Part 1 of this study (Cohorts 1-6) 
demonstrating robust, clinically meaningful reductions in liver fat assessed by MRI-PDFF and 
ALT at thresholds that have been associated with histological benefit in patients with NASH, 
combined with additional metabolic benefits, provide further support for a potential benefit in 
patients with NASH. 
Available data from clinical studies with other FGF21 analogues, including a recent Phase 2 
study with pegbelfermin in a patient population that is very similar to the intended target 
population for study BIO89-100-102, do not suggest clinically significant safety concerns. Data 
from the Phase 1 SAD study TV47948-SAD-10122 in healthy subjects suggest a good safety and 
tolerability profile with single SC doses of BIO89-100 up to 78 mg; mild injection site reactions 
and mild, sporadic GI effects were reported in BIO89-100-treated subjects. Preliminary data 
from Part 1 (Cohorts 1-6) of study BIO89-100-002 (the current study) also suggest a good safety 
and tolerability profile.  
As with any biologic drug, there is a potential risk of immunogenicity; measures have been 
implemented in this protocol to characterize this risk and appropriately identify and address any 
clinically significant events related to immunogenicity, including the potential for severe 
hypersensitivity reactions (including anaphylaxis). Bone and female reproductive findings have 
been observed at high doses in toxicology studies in mice; these risks are mitigated by the 
presence of adequate safety margins and additional precautionary measures, as detailed in the 
section on potential risks. 
Taken together, considering the benefit, potential risks, and risk mitigation measures that have 
been implemented, the sponsor considers the benefit-risk profile for administering BIO89-100 to 
patients with NASH or NAFLD at high risk for NASH and patients with biopsy-proven NASH 
(NAS ≥4 and fibrosis stage F1 with high risk, F2 or F3) in study BIO89-100-102 to be favorable. 
The overall benefit-risk associated with administration of BIO89-100 will be continually 
re-assessed with the emergence of additional data. 


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3. 
PART 1 
3.1. 
Part 1 Objectives and Endpoints 
Objectives 
Endpoints 
Primary 
• To evaluate the safety and 
tolerability of ascending 
multiple SC injections of 
BIO89-100 in subjects with 
NASH or who have NAFLD 
and at a high risk of NASH 
• Frequency and severity of AEs and 
SAEs 
• Number of subjects who discontinued 
due to AEs and due to related AEs 
• To characterize BIO89-100 PK  
• Cmax within a dosing interval 
• AUC0-tau within a dosing interval 
• tmax 
• t½ 
Additional PK parameters may be calculated if 
deemed appropriate. 
The serum concentration-time data may also be 
used for population PK modeling with the results 
reported separately from the clinical study report. 
Secondary 
• To evaluate the 
immunogenicity of BIO89-100 
as measured by presence of 
ADA 
• Assessment of the incidence and 
characteristics of ADA after dosing 
(e.g., titer and/or binding specificity, 
to the FGF21 and PEG part of BIO89-
100). 
• Impact of the presence of ADAs on 
serum BIO89-100 concentrations and 
clinical safety. 
Additional ADA assessments will be performed if 
a severe hypersensitivity reaction (e.g., 
anaphylaxis) is observed. 
• To characterize biomarkers, 
PD profile and biological 
activity of BIO89-100 
administered at ascending 
doses and with both QW and 
Q2W dosing intervals 
Change and percentage change from baseline in 
the following biomarkers/PD parameters: 
• Anthropomorphic measurements: 
− Body weight 
• Laboratory parameters: 


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Objectives 
Endpoints 
• To evaluate the time, dose, and 
exposure relationship of 
BIO89-100 on biological 
activity, as assessed by 
biomarkers and PD 
− Triglycerides 
− Non-HDL cholesterol 
− HDL-c 
− LDL-c 
− HbA1c 
− HOMA-IR 
− Liver function tests: ALT, AST  
− Adiponectin 
− Pro-C3 
− Free fatty acids and Adipo-IR (fasting 
free fatty acids  fasting insulin) 
• Imaging measures: 
− MRI-PDFF 
Other Safety Endpoints 
Exploratory Endpoints 


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Objectives 
Endpoints 
3.2. 
Part 1 Study Design 
Part 1 is a randomized, double-blind, placebo-controlled, MAD study to evaluate the safety, 
tolerability, PK and PD profiles, and immunogenicity of BIO89-100 administered SC in 
approximately 83 subjects with NASH, or with NAFLD who are at a high risk of NASH. This 
multi-site study will consist of 6 cohorts and will evaluate 2 dosing schedules, QW (Cohorts 1 to 
4) and Q2W (Cohorts 5 and 6) (Table 1). Based on emerging data and SMC recommendation, 
the Sponsor can decide to modify the dose of any subsequent cohort. 
Part 1 will include a Screening period, a Treatment period, and a Follow-up period. After signing 
informed consent, subjects will undergo screening assessments to determine eligibility over a 
period of up to 60 days. 
Eligible subjects will be randomized for each cohort as described in Section 5.4. Randomization 
to Cohorts 3 and 5 will be stratified as described in Section 5.4. The decision to randomize into 
the next cohort (dose level) will be approved by an SMC, comprised of at least a Principal 
Investigator participating in the study, the CRO Medical Monitor, and the Sponsor Medical 
Monitor. The SMC will review blinded safety data (adverse events, clinical laboratory data, vital 
signs, and ECGs), summary of PK data, if available, and other blinded relevant data. There will 
be 2 dose escalation decisions. After Cohort 1 completes the Day 36 visit, the SMC will decide 
whether subjects can be randomized into Cohorts 2 and 5 (both cohorts to start concurrently). 
After at least 8 subjects from both Cohort 2 and Cohort 5, including at least 1 subject on placebo 


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in each cohort, complete the Day 36 visit, the SMC will decide whether subjects can be 
randomized into Cohorts 3, 4, and 6 (all three cohorts to start concurrently). 
For all subjects, treatment with study intervention may be continued for 8 additional weeks (total 
of 12 weeks) following review of individual safety data (AEs, ECGs, physical examination, and 
safety laboratory) through Day 30 (4 weeks) by the site Investigator, CRO Medical Monitor, and 
Sponsor Medical Monitor. Subjects who experience clinically significant TEAEs that are 
assessed as a potential risk to subject safety will be discontinued from study intervention and 
undergo an early termination visit. The decision to discontinue study intervention will be made 
by the site Investigator following discussion with the Sponsor Medical Monitor. 
Cohorts 1 to 4 (weekly regimen): On Day -1, eligible subjects will be randomized (as described 
above) and be treated with SC injection of study intervention QW starting on Day 1 through 
Day 85; subjects will be domiciled at the Phase 1 unit from Day -1 (day before 1st dose) to Day 2 
and from Day 28 (day before the 5th dose) to Day 30. Subjects will attend ambulatory clinic 
visits for dosing and evaluations per the SoA (Section 1.2.1, Table 2). On the Day 8, Day 15 and 
Day 22 ambulatory dosing visits, subjects will remain at the study site for observation for at least 
2 hours post dose. 
Cohorts 5 and 6 (every 2 weeks regimen): On Day -1, eligible subjects will be randomized (as 
described above) and be treated Q2W with SC injection of study intervention starting on Day 1 
through Day 85; subjects will be domiciled at the Phase 1 unit from Day -1 (day before 1st dose) 
to Day 2 and from Day 28 (day before the 3rd dose) to Day 30. Subjects will attend ambulatory 
clinic visits for dosing and evaluations per the SOA (Section 1.2.1, Table 4). On the Day 15 
ambulatory dosing visit, subjects will remain at the study site for observation for at least 2 hours 
post dose. On Days 64 and 78 subjects will be contacted by phone to inquire about AEs and 
concomitant medications. 
Subjects in all cohorts will be followed up on Day 92 (1 week post last dose of study 
intervention) and Day 113, 4 weeks post last dose of study intervention (End of Study visit). 
Subjects who are found to be ADA positive at the end of study visit will be followed for 3-5 
months after this visit or until stable or declining as deemed by the Medical Monitor. 
Assessments during the study period will be done as specified in the Schedule of Activities 
(SoA; Table 2 for Cohorts 1-4 and Table 4 for Cohorts 5-6). 
For each subject, the total duration of study participation will be up to 172 days (24.5 weeks) 
excluding the potential dose interruption due to the COVID-19 pandemic: 
Screening period: 
≤ 60 days (8.5 weeks) 
Treatment period: 
84 days (12 weeks) 
Follow-up period 
28 days (4 weeks) 
3.2.1. 
Scientific Rationale for Study Design 
Part 1 is a randomized, double-blind, placebo-controlled, multiple ascending dose, 4-week study 
designed as a typical dose range finding study. The primary endpoints, safety and PK, are 
characteristic for this type of study. The study will be followed by an 8-week extension, and a 
number of secondary pharmacodynamic endpoints, including change in liver fat content as 
assessed by MRI-PDFF, will be evaluated. These will provide early proof-of-concept data to 
inform dose selection in future clinical studies. 


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3.2.2. 
Rationale for Study Population 
In recent years, there has been a shift in regulatory expectations for early, Phase 1 clinical 
studies, whereby it is increasingly expected that the population enrolled into such studies would 
consist of patients who are similar to the intended target population for the drug. BIO89-100 is 
being developed for NASH with fibrosis; in line with this approach, study BIO89-100-002 will 
enroll subjects with NASH or with NAFLD and at a high risk for NASH. Subjects will be 
required to have  evidence of  ≥ 10% liver steatosis by MRI-PDFF, together with either biopsy-
proven NASH (liver biopsy within 2 years before enrollment), or at least one of the following: 
(1) Obesity with T2DM or (2) Obesity with evidence of liver injury (latter defined as either 
increased ALT and/or Fibroscan vibration-controlled transient elastography (VCTE) 
score ≥7 KPa). It is notable that central obesity by itself increases the risk for NASH in patients 
with NAFLD, and that the additional requirement for presence of T2DM, a well characterized 
risk factor for NASH and advanced fibrosis, or evidence of liver injury, is considered appropriate 
to define a population of NAFLD patients who are at high risk for NASH. This population of 
patients with biopsy-proven NASH or NAFLD with a high risk for NASH also meets regulatory 
expectations for patients to be enrolled in early proof-of-concept studies for NASH, in which 
change in liver fat as measured by MRI-PDFF is a typical endpoint. Change in liver fat as 
determined by MRI-PDFF has emerged as the leading noninvasive quantitative biomarker for 
liver fat quantification as it is accurate, reproducible, precise, has excellent inter-rater and intra-
rater reproducibility, and has a robust correlation with MRI spectroscopy-based quantification 
(the gold standard to quantify liver triglyceride content) for liver fat, with a correlation 
coefficient ranging from 0.98 to 0.99 (Tapper, 2018).The study population is thus also 
appropriate for proof-of-concept assessment of this and other pharmacodynamic endpoints. 
3.2.3. 
Justification of Treatment Duration 
The treatment duration will be 12 weeks, consisting of a 4-week study for initial safety 
assessment (primary endpoint) with an 8-week extension phase for efficacy and safety 
assessments. The study includes an individual safety checkpoint after 4 weeks of treatment, to 
ensure that only individuals with an acceptable safety profile will proceed to the 8-week 
extension phase. The study design will enable to obtain early proof-of-concept data, including 
assessment of change in liver fat (measured by MRI-PDFF) and additional biomarkers after 
12 weeks of treatment with BIO89-100. For additional information, please also refer to the 
BIO89-100 IB. 
Factors affecting the rate of liver de-fattening are not well characterized, and it is difficult to 
predict the dynamics of improvement in hepatic steatosis based on a drug’s mechanism of action. 
Studies assessing change in liver fat by MRI-PDFF, including a recent study with another 
pegylated FGF21 product, pegbelfermin, typically had readouts at 12 weeks or later (Harrison, 
2018a; Harrison, 2018b; Sanyal, 2019). Recently, ≥30% reduction in hepatic steatosis by 
MRI-PDFF at week 12 was found to be correlated with reduction in the ballooning and 
inflammation components of NAFLD Activity Score, and to be predictive of NASH resolution 
on liver biopsy at 36 weeks. In line with these considerations, the Sponsor plans to assess change 
in liver fat by MRI-PDFF and other pharmacodynamic biomarkers after 12 weeks of treatment. 


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3.2.4. 
Justification for Dose 
Six dose cohorts are planned: 4 cohorts to be dosed QW and 2 cohorts to be dosed Q2W 
(Table 1). Based on emerging data and SMC recommendation, the Sponsor can decide to modify 
the dose of any subsequent cohort. 
These dose regimens were selected based on nonclinical safety and pharmacology studies, 
interim review of PK and blinded safety data from the single ascending dose clinical study 
TV47948-SAD-10122 and PK modelling. 
 
 
 
 
 
 
The exposure at the NOAEL for female and male mice as well as for monkey are compared to 
the observed AUC(0-168) from the highest dose in Study TV47948-SAD-10122.  In determining 
the predicted safety margins for the highest dose planned in the Study BIO89-100-002 we used a 
partial fitting approach by employing PK data from Study TV47948-SAD-10122 and a 
previously developed model in monkeys. When using the predicted AUC(0 – 168) at steady state for 
the highest dose regimen, 27 mg QW, the safety margins ranged from 
 The 
predicted Cmax safety margins for the highest administered dose, 36 mg Q2W, 
 
. The safety margins for both AUC(0 – 168) at steady state and for Cmax should provide more 
than adequate coverage for subjects in Study BIO89-100-002. 
Clinical safety observations also support the dose regimens for Study BIO89-100-002. Clinical 
safety in healthy subjects is being established with single SC doses up to 78 mg in study 
TV47948-SAD-10122. The study has been recently completed; review of blinded safety data did 
not identify clinically significant safety concerns.  Notably, the starting dose in BIO89-100-002, 
3 mg QW for 13 doses, represents a total exposure of
 
 
A PK modeling and simulation framework was undertaken to estimate the systemic exposure 
anticipated with the proposed dose regimens for Study BIO89-100-002. Based on these analyses, 
across the proposed dose range, plasma concentrations (median, 2.5th and 97.5th percentiles) are 
predicted to increase proportionally with dose up to and including 27 mg QW and 36 mg Q2W. 
The maximum proposed dose is anticipated to achieve maximal suppression of key PD responses 
associated with important clinical outcomes over the dosing interval, once steady-state is 
achieved. A 3 mg QW dose is anticipated to produce close to the half-maximal response whereas 
doses greater than 27 mg, administered QW, are not anticipated to produce substantially larger 
responses over a 27 mg dose. Therefore, the QW regimens are proposed to range from 3 mg to 
27 mg, SC. Similar magnitude of maximal response is anticipated for a 36 mg dose administered 
Q2W. 
Based on these data, the dose regimens are anticipated to be safe, of sufficient duration to reach 
steady state, to maintain PD responses over the dosing intervals, and to cover the dose range 
where key PD responses are predicted to be associated with important clinical outcomes to be 


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assessed in subsequent studies. Depending on emerging study findings, the range of doses in 
study BIO89-100-102 may be modified, and other intermediate doses may be added. 
3.2.4.1. 
Justification for Weekly (QW) and Once Every Two Weeks (Q2W) Dosing 
Regimens 
Nonclinical data from 2 pharmacological studies in spontaneously diabetic cynomolgus 
monkeys, in which the effect of BIO89-100 (up to 2 mg/kg) was evaluated following either QW 
or Q2W dosing regimens, provides the rationale for the dosing regimens with BIO89-100 in the 
current study. In these nonclinical studies, statistically significant reductions were observed in 
body weight, food intake, glucose, LDL, triglycerides, HbA1c and ALT along with improvement 
in oral glucose test results and an increase in adiponectin levels in monkeys treated with QW or 
Q2W regimens of BIO89-100, compared to the vehicle group. The exposure profile of the once 
QW and Q2W regimens was similar, except for the expected higher trough levels on Day 14 and 
Day 28 with the QW dose. As the half-life of BIO89-100 in these monkeys was approximately 
50 hours, the findings suggest the absence of a direct PK-PD correlation. 
3.2.5. 
End of Study Definition 
A subject is considered to have completed the study if he or she has completed all study periods 
including the End of Study (EOS)/Day 113 follow-up visit. The end of the study is defined as the 
date of the last visit of the last subject in the study. 
3.3. 
Part 1 Eligibility Criteria 
3.3.1. 
Part 1 Inclusion Criteria 
Subjects are eligible to be included in the study only if all of the following criteria apply: 
Age and Gender 
1. Subjects must be 21 to 75 years of age inclusive, at the time of signing the informed 
consent form (ICF). 
2. Male or female. 
Type of Subject and Disease Characteristics 
3. Evidence of steatosis by Fibroscan and MRI-PDFF: 
a. All subjects must complete a Fibroscan with Controlled Attenuation Parameter (CAP) 
measurement. Fibroscan must be done with a subject-appropriate probe during 
screening; CAP score should be ≥300 dB/m, unless a MRI-PDFF ≥10% within the 
last 12 months is available and the current CAP score is similar to CAP score when 
previous MRI-PDFF was done. 
b. MRI-PDFF ≥10% at screening (MRI must be completed no more than 35 days prior 
to randomization) 
For the purpose of these inclusion criteria, use the following definitions: 
T2DM: History of T2DM diagnosed at least 6 months prior to screening [diagnosis based on 
documentation of fasting plasma glucose (≥126 mg/dL), plasma glucose in the 75g OGTT 
(≥200 mg/dL) or HbA1c (≥ 6.5 %)]. Subjects must have a glycated hemoglobin (HbA1C) level 


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at <9.5%, without known macrovascular or clinically significant microvascular complications, 
and – if receiving antidiabetic medications – must be on a stable dose of antidiabetic medications 
before screening (6 months for Glucagon-Like Peptide 1 (GLP-1) analogs or Dipeptidyl 
Peptidase IV (DPP-IV) antagonists; 3 months for other oral or injectable medications; 
thiazolidinediones or insulin are NOT allowed in this study). 
Central Obesity: Waist circumference of >102 cm for males, >88 cm for females, or body mass 
index (BMI) >30 kg/m2. BMI should not exceed 45 kg/m2. 
Increased ALT: ALT ≥ 40 in males, ALT ≥ 30 in females. ALT level should not exceed 
200 U/L. 
4. NASH or high risk for NASH as reflected by AT LEAST ONE of the following: 
a. Diagnosis of NASH with fibrosis (stages F1, F2 or F3), without cirrhosis, by 
percutaneous liver biopsy meeting one of the following: 
a. Performed within 24 months prior to screening  
b. For subjects who did not undergo a liver biopsy in the 24 months prior to 
screening, a liver biopsy may be performed during screening if not 
contraindicated (optional, requires pre-approval by Sponsor). For subjects who 
underwent a previous liver biopsy >24 months prior to screening that would not 
have qualified for the study based on results of the initial biopsy, there should be 
a sound clinical basis to expect different findings in a repeat biopsy based on 
medical judgment. 
b. Central obesity WITH T2DM 
c. Central obesity WITH either increased ALT and/or Fibroscan VCTE score ≥7 KPa. 
Pregnancy and Contraception 
Contraceptive use by men or women should be consistent with local regulations regarding the 
methods of contraception for those participating in clinical studies. 
5. All subjects (male or female) who are of childbearing potential must agree to use highly 
effective, double contraception (both male and female partners) during the study. Double 
contraception is defined as use of a condom by the male partner, combined with use of 
one of the following forms of highly effective contraception by the female partner:  
a. a. Oral contraceptive pills,  
b. b. depot or injectable contraceptive,  
c. c. Intrauterine device (IUD),  
d. d. Contraceptive patch (e.g. Xulane®) or NuvaRing®, or  
e. e. documented evidence of surgical sterilization at least 6 months prior to screening 
visit (i.e. tubal ligation or hysterectomy).  
Use of a condom in a male subject who underwent vasectomy is also acceptable as double 
contraception. Use of highly-effective, double contraception must continue for 30 days or 5 half-
lives (whichever is longer) after the last dose of investigational product. Female subjects should 
not donate oocytes during this time. Male subjects must not donate sperm during this time. 
Rhythm methods are not considered as highly-effective methods of birth control. Subject 
abstinence for the duration of the study and 30 days or 5 half-lives (whichever is longer) after 
last dose of investigational product is acceptable if it is the subject’s regular practice. 


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6. Females of childbearing potential must have a negative serum pregnancy test at 
Screening and a negative urine pregnancy test on Day -1. Female of childbearing 
potential must agree to undergo a pregnancy test prior to each administration of study 
intervention. 
7. Females not of childbearing potential will be defined for this study as postmenopausal 
(defined as cessation of regular menstrual periods for at least 12 months) and confirmed 
by follicle stimulating hormone (FSH) level OR sterile. 
Medical documentation indicating that the subject had undergone a surgical sterilization 
procedure will be required from female study subjects to be considered not of 
childbearing potential in this study. 
Presumptive fertile male subjects who are partnered with women who are not of 
childbearing potential, either post-menopausal or through sterilization or other surgical 
procedures (e.g., hysterectomy) will confirm that their partners have been determined to 
be sterile or post-menopausal based on medical judgment; this can be confirmed either by 
statement or medical records, if presented by the subject. 
Men will not be required to use other contraception when sexually active with a sterile or 
post-menopausal partner. 
Men who are sexually active with fertile women need to confirm that they will use 
2 forms of contraception as per protocol. 
Informed Consent 
8. Capable of giving signed informed consent which includes compliance with the 
requirements and restrictions listed in the ICF and in this protocol. 
3.3.2. 
Part 1 Exclusion Criteria 
Subjects are excluded from the study if any of the following criteria apply: 
Medical Conditions 
1. History of, or current symptoms of, cardiovascular disease or cerebrovascular disease 
including clinical congestive heart failure (CHF), symptomatic coronary artery disease 
(CAD), peripheral arterial disease, and/or abdominal aortic aneurysm (AAA), history of 
transient ischemic attack (TIA), peripheral vascular disease (PVD), or symptomatic 
carotid stenosis. 
2. Clinically significant respiratory; hepatic (other than NAFLD or NASH); renal; 
gastrointestinal; neurological; immunological; hematologic, infectious or psychiatric 
disorder(s) (e.g. schizophrenia, generalized anxiety disorder, panic disorder, etc.); or a 
history of any illness that, in the opinion of the Investigator, might confound the results 
of the study, or pose additional risk to the subject by participation in the study. Individual 
cases which the Investigator deems the subject appropriate for inclusion despite a chronic 
medical condition should be discussed with and approved by the Medical Monitor. 
Subjects who are known to have tested positive for COVID-19 will be excluded, even 
they are asymptomatic. 
The following conditions are NOT EXCLUDED: 


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• Hypertension: Subjects with a history of hypertension, whose hypertension is 
controlled and are clinically stable, may be enrolled if they have been on a stable dose 
of no more than 2 antihypertensive medications at least 2 months before screening. 
• Dyslipidemia: Subjects with dyslipidemia, who are clinically stable, may be enrolled, 
including subjects who have been on a stable statin dose at least 2 months before 
screening. 
• T2DM: Subjects with T2DM may be enrolled if their HbA1c level does not exceed 
9.5 %, there are no known macrovascular or clinically significant microvascular 
complications and they have been on a stable dose of antidiabetic medications in the 6 
months before screening (for GLP-1 agonists or DPP-IV antagonists) or 3 months for 
other oral or injectable treatments. Subjects treated with thiazolidinediones or insulin 
are excluded from this study. 
• Depression: Subjects with stable, controlled depression, who have not been 
hospitalized in the past for depression and have been on a stable dose of no more than 
one antidepressant in the 2 months before screening, may be enrolled. 
3. Greater than 40% increase in ALT or AST between 2 screening assessments, to be done 
at least 2 weeks apart, as per the table below: 
ALT/AST Screening Assessments 
Eligibility status 
Assessment 1 
Assessment 2  
Assessment 3 (if applicable) 
Normal 
Normal 
Not applicable 
Eligible 
Normal  
Abnormal and ≤40% 
increase from Assessment 
1 
Not applicable 
Eligible 
Normal  
Abnormal and >40% 
increase from Assessment 
1 
Normal or ≤40% increase 
from Assessment 1 
Eligible 
Abnormal and >40% increase 
from Assessment 1 
Excluded 
Abnormal  
≤40% increase from 
Assessment 1 
Not applicable 
Eligible 
Abnormal 
>40% increase from 
Assessment 1 
≤40% increase from 
Assessment 1 
Eligible 
>40% increase from 
Assessment 1 
Excluded 
Normal is defined as ≤ ULN; abnormal is defined as >ULN  
Note: Clinical judgment should be used for subjects with isolated AST increases in whom there is suggestion of 
another cause of AST increase (e.g. muscle injury as evident by concurrent creatine phosphokinase elevation). 
4. A personal or family history of arrhythmia, sudden unexplained death at a young age 
(before 40 years) in a first-degree relative, or long QT syndrome. 
5. History of bariatric surgery or plan to have bariatric surgery during conduct of study. 
6. History of type 1 diabetes. 


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7. Weight loss of more than 5% within 3 months prior to Day -1 or more than 10% within 
6 months prior to Day -1 or planning to try to lose weight during conduct of study. 
8. History of a liver disorder other than NASH or clinical suspicion of a liver disorder other 
than NASH, including but not limited to hepatitis B and hepatitis C, autoimmune 
hepatitis, hemochromatosis, alcoholic liver disease, primary sclerosing cholangitis (PSC), 
primary biliary cirrhosis (PBC), or Wilson’s disease. 
9. History of liver transplantation. 
10. History of cirrhosis or evidence of cirrhosis (clinical, imaging-liver appearance or 
splenomegaly), advanced fibrosis (F4) on biopsy, VCTE-based Fibroscan >12.5 kPa or 
Fibrosis-4 (Fib-4) index5 >3.25. In cases in which a patient is determined to have stage 4 
fibrosis based on VCTE or FIB4, but has a liver biopsy within 12 months of screening 
and reported to show fibrosis  stage F0, F1, F2 or F3, the subject will be considered to be 
eligible based on the liver biopsy result.  
11. Major trauma or surgery in the 2 months before screening or at any time between 
screening and Day -1. 
12. Recent clinically significant acute illness (within 4 weeks of screening) unless per the 
Investigator’s clinical discretion a full recovery is apparent. 
13. History of bone trauma, fracture or surgery within 2 months of screening. 
14. Have any known malignancy or history of malignancy, except for basal cell skin cancer 
that has been treated with no evidence of recurrence for at least 3 months prior to Day -1. 
In specific circumstances in which there is basis to consider the subject as cured from 
cancer, exceptions may be considered. Any exception would need to be approved in 
advance by the Sponsor’s Medical Monitor. 
15. History of alcohol use disorder (per DSM-5) or risky drinking (defined as alcohol intake 
>14 standard drink units per week or 4 standard drinks on a single occasion in men; and 
alcohol intake >7 standard drink units per week or 3 standard drinks on a single occasion 
in women) within the 24 months before Day -1, or had a positive alcohol test at screening 
and/or check-in. A unit of alcohol is defined as 14 gram or as 355 mL of beer (5%), 
1 glass of wine (150 mL; 12%), or 1 shot of hard liquor (45 mL; 40%). During the study, 
subjects will be encouraged to abstain from alcohol. Alcohol intake will be limited to 
2 drinks per day for men and 1 drink per day for women. 
16. History of drug abuse, or any other substance dependence (with the exception of caffeine) 
as defined by the DSM-IV-TR in the past 2 years prior to screening or a positive test for 
drugs of abuse (i.e., amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine 
metabolites, opiates, and phencyclidine) at screening and/or Day -1. 
17. Daily use of more than 10 cigarettes/day, or 2 cigars/day, or equivalent use of any 
tobacco product within 6 weeks prior to Screening. E-cigarettes or other vaping devices 
should not deliver more than 15 mg of nicotine/day. 
18. Pregnant or breastfeeding, or planning to become pregnant or breastfeed while enrolled in 
the study or within 30 days or 5 half-lives (whichever is longer) after last dose of study 
 
5 FIB4 score = Age × AST (IU/l)/platelet count (× 109/l) × √ALT (IU/l) 


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intervention. Presumptive fertile, sexually active male subjects, whose female partner is 
pregnant, will be excluded.  
Prior/Concomitant Therapy 
19. Subject report of use of medications historically associated with secondary NAFLD for 
more than 2 consecutive weeks in the 12 months prior to screening (e.g., amiodarone, 
methotrexate, systemic glucocorticoids, tetracyclines, tamoxifen, estrogens at doses 
greater than those used for hormone replacement, anabolic steroids, valproic acid, and 
other medications with known hepatotoxicity). 
20. Any prior exposure to a FGF21 analogue (e.g. BIO89-100, LY2405319, LY3025876, 
BMS-986036, BMS-986171, PF-05231023, PF-06645849, AKR-001) or FGFR1 
activating product, if known. 
21. Any previous long-term (> 4 weeks) use of systemic steroid (glucocorticoid) medications 
such as prednisone. 
22. Vitamin E supplementation beyond Recommended Dietary Allowance (22.4 IU/day) 
within 6 months before screening. 
23. Any systemic medications, including over-the-counter (OTC) medications, prescription 
drugs, biologics, vaccines, herbal preparations, used within 14 days, or 5 half-lives, 
whichever is longer, before Day -1 and throughout the study with the FOLLOWING 
EXCEPTIONS: 
a. Acetaminophen (up to 4 g/day, use as indicated in the label) 
b. Ibuprofen (up to 1.2 g/day of ibuprofen use as indicated in the label) 
c. Aspirin (up to 81 mg/day) 
d. Antihypertensive medications (no more than 2 agents) on stable dose for at least 
2 months prior to screening 
e. Antidiabetic medications: stable dose for 6 months prior to screening for GLP-1 
analogs or DPP-IV antagonists; 3 months prior to screening for oral or other 
injectable medications. Thiazolidinendiones and insulin are not allowed in this study. 
f. Statins on stable dose for at least 2 months 
g. Asthma treatment with exception of oral corticosteroids 
h. Anti-depressants (no more than 1 agent) on stable dose for at least 2 months 
i. Other exceptions must be approved by the Medical Monitor 
j. Vitamins (other than vitamin E at a dose greater than recommended dietary 
allowance) and food supplements are allowed if the subject had been taking a stable 
regimen for at least 2 months before screening. 
Prior/Concurrent Clinical Study Experience 
24. Any investigational drug small molecule (new chemical entity) within 30 days and large 
molecule (biologics) within 90 days, or 5 half-lives, whichever is longer, prior to Day -1, 
if known. 
Diagnostic Assessments 
25. Any history of suicidal behavior or suicidal ideation with plan and intent based upon 
clinical history. A “yes” response to any of the suicidal ideation or suicidal behavior 
questions in the C-SSRS on Day -1. 


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26. Any clinically significant laboratory abnormality at screening. One repeat test may be 
allowed at the discretion of the Investigator. The presence of one or more of the 
following laboratory abnormalities should lead to exclusion of the subject from 
participating in the study: 
a. ALT or AST ≥ 200 U/L 
b. Elevation of total bilirubin (TB) > ULN 
c. Normalized ratio (INR) > ULN 
d. Alkaline phosphatase at screening >1.25 ULN 
e. Glomerular filtration rate (eGFR) ≤60 mL/min/1.73 m2 as estimated by CKD-EPI 
Creatinine equation (Levey, 2009) 
f. Serum total triglycerides ≥ 1000 mg/dL 
g. HbA1c ≥ 9.5% 
27. ECG abnormality that may, in the opinion of the Investigator, interfere with study 
participation, including intraventricular conduction delays (QRS interval ≥120 msec or 
PR interval >200 msec (220 msec in individuals with a heart rate <70 beats per minute), 
and resting QTcF interval of ≤320 msec and/or ≥450 msec for males or ≥470 msec for 
females. 
28. Vital sign abnormalities on screening, including: having a supine systolic blood pressure 
<90 or >150 mmHg, diastolic blood pressure (following at least a 5 minute rest) <50 or 
>95 mmHg, or resting heart rate <50 or >100 beats per minute. 
29. BMI at screening <25 or ≥ 45 kg/m2. 
Other Exclusions 
30. Known allergy or sensitivity to injected proteins, or any component of the formulation, or 
has a history of allergies (including food allergies) requiring acute or chronic treatment 
(except for seasonal allergic rhinitis). 
31. Donated or received any blood or blood products (e.g., white blood cells, platelets) within 
the 60 days prior to screening, or has donated blood or blood products at least twice 
within the 6 months prior to screening, or the subject has donated plasma within 7 days of 
the screening visit, or has planned donations during the 56 days or 5 half-lives following 
the last day of study intervention administration, whichever is longer. 
32. Any abnormality of the skin or abdominal wall that would affect SC administration to the 
abdominal area or any tattoos or scars in the intended injection area. 
33. An employee of the investigational center or has a family member who is involved with 
the conduct of this study. 
34. Subject who cannot undergo MRI for any reason (e.g., contraindication, claustrophobia, 
excessive weight or body size for MRI machine). 
35. Subject who cannot fast for study procedures for any reason. Specifically, subjects with 
T2DM who have had one or more episodes of hypoglycemia or past issues with fasting 
will be excluded. Subjects with T2DM who are treated by insulin secretagogues will need 


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to consult their treating physician about the optimal timing to take these medications to 
enable them to fast safely for study procedures. 


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4. 
PART 2 
4.1. 
Part 2 Objectives and Endpoints 
Objectives 
Endpoints 
Primary 
• To evaluate the safety and 
tolerability of SC injections of 
27 mg BIO89-100, 
administered weekly for 20 
weeks, in subjects with biopsy-
proven NASH (NAS ≥4, 
fibrosis stage F1 with high 
risk, F2 or F3)   
• Frequency of AEs and SAEs 
• To characterize effect of 
BIO89-100 on liver histology  
• At least a 2-point improvement in 
NAFLD Activity Score (NAS) with at 
least a 1-point improvement in 
ballooning or lobular inflammation, 
and no worsening of fibrosis 
Secondary 
• To characterize biomarkers, 
PD profile, and biological 
activity of BIO89-100  
Change and percentage change from baseline in 
the following biomarkers/PD parameters: 
• Anthropomorphic measurements: 
− Body weight 
• Laboratory parameters: 
− Triglycerides 
− non-HDL cholesterol 
− HDL-c 
− LDL-c 
− HbA1c 
− Liver function tests: ALT, AST 
− Pro-C3 
• Imaging measures: 
− MRI-PDFF 
• To characterize effect of 
BIO89-100 on liver histology 
• Improvement of fibrosis ≥1 stage 
without worsening of NASH 


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Objectives 
Endpoints 
• NASH resolution without worsening 
of fibrosis6 
• To characterize BIO89-100 PK  
• Trough concentration of BIO89-100 
Other Safety Endpoints 
Exploratory Endpoints 
 
6 Resolution of NASH includes the total absence of ballooning (score=0) and absent or mild inflammation (score 0 
to 1) associated with at least a 2-point reduction in NAS, and no worsening if fibrosis (progression ≥ 1 stage). 


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Objectives 
Endpoints 
 
7 ≥30% relative reduction in liver fat by MRI-PDFF at D85 
8 Resolution of NASH includes the total absence of ballooning (score=0) and absent or mild inflammation (score 0 
to 1) associated with at least a 2-point reduction in NAS, and no worsening if fibrosis (progression ≥ 1 stage). 


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Objectives 
Endpoints 
4.2. 
Part 2 Study Design 
Part 2 is an open-label cohort that will enroll 20 subjects with biopsy-proven NASH and fibrosis 
(NAS ≥4, fibrosis stage F1 with high risk, F2 or F3). Subjects will either undergo a liver biopsy 
during screening or have a recent liver biopsy (within ≤24 weeks before Day 1, available to be 
evaluated for eligibility by the central reader) that meets study inclusion criteria. Eligible 
subjects will be treated with weekly (QW) SC injection of study intervention starting on Day 1 
and continuing through Day 134 (20 weeks of treatment). Subjects will attend ambulatory clinic 
visits for dosing and evaluations and home visits for dosing9 as per the Schedule of Activities 
(SoA; Section 1.2.2, Table 6). Subjects will undergo a second liver biopsy within 14 days after 
the last dose of study intervention. Subjects will be followed up on Day 141 (1 week post last 
dose of study intervention) and Day 162 (4 weeks post last dose of study intervention, End of 
Study visit). Subjects who are found to be NAb-positive at the End of Study visit will be 
followed approximately every 3 months starting from when the site has been notified of the 
positive result, until: (1) neutralizing antibodies are no longer detectable or (2) the subject has 
been followed for a period of at least 1 year (± 4 weeks). More frequent testing (e.g., every 
month) or testing for a longer period of time may be requested in the event of safety-related 
concerns. 
4.2.1. 
Scientific Rationale for Part 2 Study Design 
Part 2 is designed as an open-label cohort. Preliminary data from placebo-controlled Part 1 
demonstrated favorable safety and tolerability with repeated dosing of BIO89-100 for 12 weeks. 
In addition, in Part 1, robust, clinically meaningful reductions in liver fat assessed by MRI-PDFF 
and ALT, at thresholds that have been associated with histological benefit in patients with 
NASH, have been observed. Part 2 extends the study to subjects with biopsy-proven NASH and 
fibrosis (NAS ≥4, fibrosis stage F1 with high risk, F2 or F3), similar to the population that is 
typically enrolled in later phase NASH studies, who will be treated with BIO89-100 at a dose of 
27 mg QW for 20 weeks. In addition to the primary safety endpoints, this will be the first study 
to characterize the effect of BIO89-100 on liver histology. Many of the same secondary PD 
endpoints as in Part 1 will be evaluated, including change in liver fat content as assessed by 
MRI-PDFF, to inform study design and choice of endpoints in future clinical studies. 
 
9 If home administration is not feasible, not desired by the subject, or deemed inappropriate option by the 
investigator for any reason, in-clinic study intervention administration may take place at some or all timepoints. 


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4.2.2. 
Rationale for Part 2 Study Population 
BIO89-100 is being developed for NASH with fibrosis. Preliminary data from Part 1 
demonstrated favorable safety and tolerability and robust, clinically meaningful reductions in 
liver fat assessed by MRI-PDFF and in ALT, with reductions in both measures beyond 
thresholds that have been associated with histological benefit in patients with NASH, with 
repeated dosing of BIO89-100 for 12 weeks, in a similar population to the intended population in 
Part 2 . Part 2 extends the study to subjects with biopsy-proven NASH and fibrosis (NAS ≥4, 
fibrosis stage F1 with high risk, F2 or F3). This population of patients meets current regulatory 
expectations for patients to be enrolled in later stage studies for NASH, and will allow to assess 
changes in liver histology, as well as changes in liver fat as measured by MRI-PDFF and other 
biomarkers in a population of patients aligned with NASH registration studies. Histological 
endpoints will include at least a 2-point improvement in NAS with at least a 1-point 
improvement in ballooning or lobular inflammation with no worsening of fibrosis (primary), 
improvement of fibrosis by at least 1 stage without worsening of NASH, and NASH resolution 
without worsening of fibrosis (secondary), and additional exploratory endpoints. In addition to 
other exploratory histological endpoints, exploratory analysis of histological endpoints in the 
subset of MRI-PDFF responders (≥30% relative reduction in liver fat by MRI-PDFF at D85) will 
be conducted. The study population is appropriate for assessment of these and other PD 
endpoints. 
4.2.3. 
Justification for Part 2 Treatment Duration 
The treatment duration will be 20 weeks, with weekly (QW) doses. Favorable safety and 
tolerability of 12-weeks of dosing with BIO89-100 at a dose of 27 mg QW was shown in Part 1. 
Based on results of recently published studies of Phase 2 clinical studies in a similar population 
of NASH patients, including a study with another FGF21 analogue (AKR-001, Akero), a 
treatment duration of 20 weeks is considered appropriate for an initial evaluation of the 
histological endpoints that will be assessed in this study.  
4.2.4. 
Justification for Part 2 Dose 
Subjects in open-label Cohort 7 will be dosed with 27 mg BIO89-100 QW for 20 weeks. 
Administration of 27 mg QW to patients with NASH or NAFLD and high risk for NASH 
(phenotypic NASH) for 12 weeks has been studied in Part 1 of this study. Justification for 
administration of 27 mg BIO89-100 QW, which was the highest QW dose in Part 1, was based 
on nonclinical safety and pharmacology studies, including a 13-week interim analysis of the 
ongoing GLP 26-week toxicology study in mice (Study 6700477), PK, and safety data from the 
single ascending dose clinical study TV47948-SAD-10122 and PK modeling, and is provided in 
Section 3.2.4. Results of the GLP 26-week toxicology study in mice (Study 6700477), that has 
been completed, are available in the IB, and support 20 weeks of weekly dosing with 27 mg 
BIO89-100.    
In Part 1 of Study BIO89-100-002, 10 subjects were randomized to Cohort 4 (27 mg QW). 
Preliminary data show that, while significant absolute and relative reduction in liver fat as 
assessed by MRI-PDFF at week 13 was observed for all studied doses, maximal reduction was 
achieved with the
 
 


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It has been shown that MRI PDFF response ≥30% (Loomba, 2015; Loomba, 2017; Harrison, 
2019) and ALT reduction ≥17 U/L (Loomba, 2019) are associated with a higher rate of NASH 
resolution and fibrosis reduction. In view of this, and considering the robust, clinically 
meaningful effects on MRI-PDFF and ALT and the favorable safety and tolerability profile of 
the 27 mg QW dose, the Sponsor believes that 27 mg QW is the most appropriate dose to be 
studied in an initial investigation of the potential effect of BIO89-100 on histological endpoints 
in a clinically relevant population of patients with NASH. The results of this study are expected 
to provide valuable information to guide dose selection for future BIO89-100 studies with 
histological endpoints in NASH.   
4.2.5. 
Part 2 End of Study Definition 
A subject is considered to have completed the study if he or she has completed all study periods 
including the End of Study (EOS)/Day 162 follow-up visit. The end of the study is defined as the 
date of the last visit of the last subject in the study. 
4.3. 
Part 2 Eligibility Criteria 
4.3.1. 
Part 2 Inclusion Criteria 
Subjects are eligible to be included in the study only if all of the following criteria apply: 
Age and Gender 
1. Subjects must be 21 to 75 years of age inclusive, at the time of signing the informed 
consent form (ICF). 
2. Male or female. 
Type of Subject and Disease Characteristics 
3. Increased metabolic risk; subjects must have at least 2 of the following:  
a. Central obesity: Waist circumference of >102 cm for males, >88 cm for females, or 
body mass index (BMI) >30 kg/m2. BMI should not exceed 50 kg/m2 
b. Fasting plasma glucose ≥126 mg/dL or previously diagnosed T2DM  
c. Increased fasting triglycerides (≥150 mg/dL) or on treatment for hypertriglyceridemia  
d. Reduced fasting HDL cholesterol (<40 mg/dL for male and <50 mg/dL for female) 
e. Hypertension or treatment of previously diagnosed hypertension  


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Only subjects meeting Criterion 3 should be further evaluated for participation. Subjects 
who do not initially meet Criterion 3 may be re-tested, based on investigator judgment. 
4. Evidence of fibrosis – must meet at least one of the following criteria: 
a. Fibroscan VCTE score ≥8.5 kPa at screening or within the last 3 months10 AND 
Aspartate aminotransferase (AST) > 20 U/L for males or >17 U/L for females in the 
first screening measurement11 
b. A historical liver biopsy obtained more than 24 weeks and less than 2 years before 
Screening with Stage 1, 2 or 3 fibrosis.  
5. Evidence of steatosis by Fibroscan and MRI-PDFF: 
a. All subjects must complete a Fibroscan with CAP measurement. Fibroscan must be 
done with a subject-appropriate probe during screening; CAP score should be 
≥280 dB/m at screening or within the last 3 months unless an MRI-PDFF ≥8% within 
the last 12 months is available and the current CAP score is similar to or greater than 
CAP score when previous MRI-PDFF was done. 
NOTE: If a Fibroscan model with CAP technology is not available, 
MRI-PDFF will be adequate to assess steatosis.   
b. MRI-PDFF ≥8% at screening (MRI must be completed no more than 35 days prior to 
baseline) 
6. Biopsy-proven NASH in a liver biopsy obtained within 24 weeks of baseline with 
fibrosis stage F1, F2, or F3 and NAS ≥4, with a score of at least 1 in each of steatosis, 
ballooning degeneration, and lobular inflammation.  
Liver biopsy may be obtained during screening or be a historical percutaneous biopsy 
that is deemed suitable for interpretation by the central reader (in up to approximately 
20% of subjects12), if the patient had no significant change in metabolic status (control of 
diabetes or hyperlipidemia, >5% weight loss or gain) or medications.   
Patients with fibrosis stage F1 with high risk will be eligible (no more than approximately 
20% of subjects). Fibrosis stage F1 with high risk is defined as fibrosis stage F1 AND at 
least one of the following criteria: a. T2DM, b. BMI ≥30 kg/m2, c. ALT > 1.5 x ULN at 
screening, d. elevated Pro-C3 (>14 ng/mL) within 6 months before screening. 
Pregnancy and Contraception 
Contraceptive use by men or women should be consistent with local regulations regarding the 
methods of contraception for those participating in clinical studies. 
7. All subjects (male or female) who are of childbearing potential must agree to use highly 
effective, double contraception (both male and female partners) during the study. Double 
 
10 If subject had fibroscan a VCTE score ≥ 8.5 kPa within last 3 months, screening VCTE score should be ≥ 7.5 kPa.  
11 If there is reason to suspect contribution of a non-hepatic source of AST (e.g., in the presence of elevated. creatine 
phosphokinase [CPK] or other evidence of muscle injury), AST should be re-tested when there is no longer evidence 
of potential contribution of an extra-hepatic source. The first assessment for Exclusion criterion 3 will be the one 
that made the subject eligible according to Inclusion criterion 4. 
12 Subjects with an eligible historical biopsy that was obtained within 24 weeks from baseline do not need to meet 
Inclusion Criteria 4 and 5. 


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contraception is defined as use of a condom by the male partner, combined with use of 
one of the following forms of highly effective contraception by the female partner:  
a. Oral contraceptive pills,  
b. Depot or injectable contraceptive,  
c. Intrauterine device (IUD),  
d. Contraceptive patch (e.g. Xulane®) or NuvaRing®, or  
 
Use of a condom in a male subject who underwent vasectomy is also acceptable as double 
contraception. Use of highly effective, double contraception must continue for 30 days or 
5 half-lives (whichever is longer) after the last dose of study intervention. Female subjects 
should not donate oocytes during this time. Male subjects must not donate sperm during this 
time. Rhythm methods are not considered as highly effective methods of birth control. Subject 
abstinence for the duration of the study and 30 days or 5 half-lives (whichever is longer) after the 
last dose of study intervention is acceptable if it is the subject’s regular practice. 
8. Females of childbearing potential must have a negative serum pregnancy test at 
Screening and a negative urine pregnancy test on Day 1. Females of childbearing 
potential must agree to undergo a pregnancy test prior to each administration of study 
intervention. 
9. Females not of childbearing potential will be defined for this study as postmenopausal 
(defined as cessation of regular menstrual periods for at least 12 months) and confirmed 
by follicle-stimulating hormone (FSH) level OR surgically sterile. For the purposes of the 
study, surgical sterilization includes hysterectomy, bilateral salpingectomy, or bilateral 
oophorectomy or bilateral tubal occlusion. 
10. Presumptive fertile male subjects who are partnered with women who are not of 
childbearing potential, either postmenopausal or through sterilization or other surgical 
procedures (e.g., hysterectomy) will confirm that their partners have been determined to 
be sterile or postmenopausal based on medical judgment; this can be confirmed either by 
statement or medical records, if presented by the subject. Men will not be required to use 
other contraception when sexually active with a sterile or postmenopausal partner. 
11. Men who are sexually active with women of childbearing potential need to confirm that 
they will use 2 forms of contraception as per protocol. 
Informed Consent and Study Requirements 
12. Capable of giving signed informed consent, which includes compliance with the 
requirements and restrictions listed in the ICF and in this protocol.  
13. Subjects must be available for the entire duration of the study and be willing to undergo 
2 liver biopsies, one at screening (unless a qualifying historical biopsy is available within 
24 weeks before baseline, in which case subjects will undergo only the second liver 
biopsy) and a second biopsy the end of the study. 
14.  Subjects must not participate in any other clinical trial throughout the duration of this 
study. Coronavirus virus disease 2019 (COVID-19) protocols may be excepted with 
Medical Monitor (or designee) approval. 


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4.3.2. 
Part 2 Exclusion Criteria 
Subjects are excluded from the study if any of the following criteria apply: 
Medical Conditions 
1. Unstable or clinically significant cardiovascular or cerebrovascular disease: 
a. Unstable angina, myocardial infarction, coronary artery bypass graft (CABG) 
surgery, percutaneous coronary intervention (PCI), transient ischemic attack (TIA) or 
cerebrovascular accident (CVA) within 6 months prior to Screening. 
b. Symptomatic valvular or other structural heart disease. 
c. Symptomatic congestive heart failure. 
d. Symptomatic, uncontrolled or high-risk arrhythmia or genetic predisposition to high-
risk arrhythmia in the subject or a first degree relative. 
e. Implanted defibrillator or pacemaker. 
f. High risk abdominal aortic aneurysm, uncontrolled peripheral vascular disease, or 
symptomatic carotid stenosis. 
2. Any other clinically significant disorder or prior therapy that, in the opinion of the 
Investigator, might pose additional risk to the subject, make the subject unable to comply 
with the protocol requirements, or confound the results of the study.  Individual cases in 
which the Investigator deems the subject appropriate for inclusion despite a clinically 
significant chronic medical condition should be discussed with and approved by the 
Medical Monitor. Subjects who have tested positive for coronavirus virus disease 2019 
(COVID-19) within 3 months prior to screening will be excluded, even if they are 
asymptomatic. 
3. Greater than 40% increase in ALT or AST between 2 screening assessments, to be done 
at least 2 weeks apart, as per the table below: 
ALT/AST Screening Assessments 
Eligibility Status 
Assessment 1 
Assessment 2  
Assessment 3 (if applicable) 
Normal 
Normal 
Not applicable 
Eligible 
Normal  
Abnormal and ≤40% 
increase from Assessment 
1 
Not applicable 
Eligible 
Normal  
Abnormal and >40% 
increase from Assessment 
1 
Normal or ≤40% increase 
from Assessment 1 
Eligible 
Abnormal and >40% increase 
from Assessment 1 
Excluded 
Abnormal  
≤40% increase from 
Assessment 1 
Not applicable 
Eligible 
Abnormal 
>40% increase from 
Assessment 1 
≤40% increase from 
Assessment 1 
Eligible 
>40% increase from 
Assessment 1 
Excluded 
Normal is defined as ≤ ULN; abnormal is defined as >ULN.  


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Note: Clinical judgment should be used for subjects with isolated AST increases in whom there is suggestion of 
another cause of AST increase (e.g., muscle injury as evident by concurrent creatine phosphokinase elevation).  
4. History of bariatric surgery within the prior 5 years (including gastric banding or sleeve 
surgery) or plan to have bariatric surgery during conduct of study. 
5. History of type 1 diabetes.  
6. Weight loss of more than 5% within 3 months prior to Day 1 or more than 10% within 
6 months prior to Day 1. 
7. History of a liver disorder other than NASH or clinical suspicion of a liver disorder other 
than NASH, including but not limited to hepatitis B and hepatitis C, autoimmune 
hepatitis, hemochromatosis, alcoholic liver disease, PSC, PBC, alpha 1-antitrypsin 
deficiency, or Wilson’s disease. Subjects who were previously diagnosed with chronic 
HCV infection who achieved sustained viral response (SVR) following treatment with a 
direct-acting antiviral (DAA) regimen at least 2 years prior to screening are not excluded. 
8. Prior or planned liver transplantation. 
9. History of cirrhosis or evidence of cirrhosis (clinical, imaging-liver appearance or 
splenomegaly), advanced fibrosis (F4) on biopsy, VCTE-based Fibroscan >20.0 kPa. In 
cases in which a patient is determined to have Stage 4 fibrosis based on VCTE but has an 
eligible liver biopsy within 6 months of Day 1, the subject will be considered to be 
eligible based on the liver biopsy result.  
10. History of bone trauma, fracture, or surgery within 2 months of screening or other bone 
disorders, such as osteoporosis, osteomalacia, or known, untreated severe vitamin D 
deficiency (serum 25-hydroxy-vitamin D ≤5 ng/mL).  
11. Have any known malignancy or history of malignancy, except for basal cell skin cancer 
that has been treated with no evidence of recurrence for at least 3 months prior to Day 1. 
In specific circumstances in which there is basis to consider the subject as cured from 
cancer, exceptions may be considered. Any exception would need to be approved in 
advance by the Sponsor’s Medical Monitor. 
12.  Current or history of significant alcohol consumption for a period of more than 3 
consecutive months within 1 year prior to screening. Defined as more than 14 units/week 
for females and more than 21units/week for males on average. 
13. History of drug abuse, or any other substance dependence (with the exception of caffeine) 
as defined by the DSM-IV-TR in the past 2 years prior to screening. Subjects without a 
history of substance use disorder who have a positive drug screen test at screening may 
be considered for enrollment with Medical Monitor (or designee) approval. Chronic, 
habitual use of cannabis/cannabinoids is exclusionary. 
14. Pregnant or breastfeeding, or planning to become pregnant or breastfeed while enrolled in 
the study or within 30 days or 5 half-lives (whichever is longer) after last dose of study 
intervention. Presumptive fertile, sexually active male subjects, whose female partner is 
pregnant, will be excluded.  


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Prior/Concomitant Therapy 
15. Subject report of use of medications historically associated with secondary NAFLD for 
more than 2 consecutive weeks in the 12 months prior to screening (e.g., amiodarone, 
methotrexate, systemic glucocorticoids [>2 weeks of pharmacologic dosing], 
tetracyclines, tamoxifen, estrogens at doses greater than those used for hormone 
replacement, anabolic steroids, valproic acid, and other medications with known 
hepatotoxicity). Short term oral steroid bursts with/without taper as well as topical and 
inhaled steroids are allowed. 
16. Any prior exposure to a FGF21 analogue (e.g., BIO89-100, LY2405319, LY3025876, 
BMS-986036, BMS-986171, PF-05231023, PF-06645849, AKR-001) or FGFR1 
activating product, if known. 
17. Any investigational drug small molecule (new chemical entity) within 30 days and large 
molecule (biologics) within 90 days, or 5 half-lives, whichever is longer, prior to Day 1, 
if known. 
18. Subjects taking Vitamin E must be on a stable dose for at least 6 months prior to the 
qualifying liver biopsy (historical or screening). 
19. Subjects may be enrolled if they have been on a stable anti-diabetic regimen for 3 months 
prior to biopsy (historical or screening). Stable regimen is defined as no addition or 
discontinuation of antidiabetic medications, but dose adjustments per standard of care are 
allowed. Insulin and DPP-IV antagonists are allowed if subject has been on a stable 
regimen for at least 3 months. GLP-1 agonists and SGLT2 inhibitors are allowed if 
subject has been on a stable dose for 6 months. Thiazolidinediones are not allowed. 
20. Subjects on antiplatelet or anticoagulant medications in whom there is a significant 
medical risk to stopping treatment with these medications temporarily (as required per 
local practice for liver biopsy) due to the nature of the treated medical condition should 
be excluded.  
Diagnostic Assessments 
21. Suicidal behavior or suicidal ideation with plan and intent based: A “yes” response to any 
of the suicidal ideation or suicidal behavior questions in the C-SSRS within the 12 
months prior to Day 1. 
22. Any clinically significant laboratory abnormality at screening including but not limited to 
the following. One repeat test may be allowed at the discretion of the Investigator.  
g. ALT or AST ≥250 U/L 
 
 
 
 
 
 
 
 
  
 


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23. ECG abnormality that may, in the opinion of the Investigator, interfere with study 
participation or present potential significant patient safety risk.  Resting QTcF interval 
≥450 msec for males or ≥470 msec for females. 
24. Clinically significant vital sign abnormalities on screening, including having a systolic 
blood pressure <90 or >150 mmHg, diastolic blood pressure (following at least a 
5 minute rest) <50 or >95 mmHg. 
25. BMI at screening <25 or ≥ 50 kg/m2. 
Other Exclusions 
26. History of anaphylaxis or known allergy or sensitivity to any component of the study 
drug. 
27. Any abnormality of the skin or abdominal wall that would affect SC administration to the 
abdominal area. 
28. An employee of the investigational center or has a family member who is involved with 
the conduct of this study. 
29. Inability to undergo a liver biopsy safely for any reason 
30. Subject who cannot undergo MRI for any reason (e.g., contraindication, claustrophobia, 
excessive weight or body size for MRI machine). 
31. Subject who cannot fast for study procedures for any reason. Specifically, subjects with 
T2DM who have had one or more episodes of hypoglycemia or past issues with fasting 
will be excluded. Subjects with T2DM who are treated by insulin secretagogues will need 
to consult their treating physician about the optimal timing to take these medications to 
enable them to fast safely for study procedures. 
 


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5. 
STUDY INTERVENTION 
Study intervention is defined as BIO89-100 (or matching placebo in Part 1), intended to be 
administered to a study subject according to the study protocol. 
5.1. 
Study Intervention(s) Administered 
Study intervention will be administered SC to the abdomen region by qualified study personnel. 
In Part 2, some study intervention administrations may occur in the subject’s home13. 
ARM  
Active arm 
Placebo arm (control) 
Intervention  
BIO89-100  
Matching placebo  
Type  
Biologic  
Chemical solution 
Dose 
Formulationa 
Each 0.7 mL vial contains 18 mg of 
BIO89-100 in Tris buffer solution 
(containing sodium chloride, sucrose, 
and polysorbate 20) at a concentration 
of 26 mg/mL  
Each 0.7 mL vial contains Tris buffer 
solution (containing sodium chloride, 
sucrose, and polysorbate 20) 
Unit Dose 
Strength(s) 
mg 
Not applicable 
Dosage Level(s)b 
Part 1 
3 mg, 9 mg, 18 mg, 27 mg weekly 
(QW) 
18 mg and 36 mg every other week 
(Q2W) 
Note: Based on emerging data and 
SMC recommendation, the Sponsor can 
decide to modify the dose of any 
subsequent cohort 
Part 1 
NA; matching placebo will be injected 
at matching frequency per assigned 
cohort. 
Part 2 
27 mg weekly (QW) 
Part 2 
Not applicable 
Route of 
Administration 
SC injection 
SC injection 
Sourcing 
Provided centrally by the Sponsor 
Provided centrally by the Sponsor 
Packaging and 
Labeling 
BIO89-100 is supplied as a sterile, 
preservative-free, frozen formulation in 
a single-use Type 1 clear glass vial for 
SC injection. Each vial will be labeled 
as required per country requirement. 
Placebo will be supplied as a sterile, 
preservative-free, frozen formulation in 
a single-use Type 1 clear glass vial for 
SC injection. Each vial will be labeled 
as required per country requirement. 
a Note, in the original batch: each 1.5 mL vial contains 36 mg of BIO89-100 
b The actual doses will be ±5% the mg dose due to technical considerations related to drug withdrawal from the 
vials into the syringes for injection. This difference is considered negligible for subject exposure. 
 
13 Details on home administration are provided in the Home Administration Manual. If home administration is not 
feasible, not desired by the subject, or deemed inappropriate by the investigator for any reason, in-clinic study 
intervention administration may take place at some or all of the designated timepoints.  


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5.2. 
Administration Instructions 
The SC injection of the study intervention should be performed by a limited number of site staff 
specifically trained in the administration of SC injections. However, administration of study 
intervention by a trained home health care worker will be considered on a case-by-case basis if 
circumstances related to the COVID-19 pandemic preclude dosing at the study site. Injections 
will be administered in the abdominal area only. Details on injection of study intervention will be 
provided in the relevant study manual. 
After administration of the study intervention, the subject should remain supine for 
approximately 15 minutes. For additional doses in the same subject, the injection site should be 
rotated within the abdominal area by at least 2 inches. Injections should be at least 2 inches from 
the umbilicus (belly button). 
In Part 2, SC injection of the study intervention will be performed by qualified site staff when the 
subject attends clinic visits. Details on self-administration at home, including training 
requirements, are provided in the Home Administration Manual. 
5.3. 
Preparation/Handling/Storage/Accountability 
The Investigator or designee must confirm appropriate temperature conditions have been 
maintained during transit for all study intervention received and any discrepancies are reported 
and resolved before use of the study intervention. 
Only subjects enrolled in the study may receive study intervention and only authorized site staff 
may supply or administer study intervention during in-clinic visits with the exception that 
administration of study intervention by a trained home health care worker will be considered on 
a case-by-case basis if circumstances related to the COVID-19 pandemic preclude dosing at the 
study site. In Part 2, study subjects may self-administer study intervention either in the clinic 
(under supervision) or at home. All study intervention 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. 
The Investigator, institution, or the head of the medical institution (where applicable) is 
responsible for study intervention accountability, reconciliation, and record maintenance (i.e., 
receipt, reconciliation, and final disposition records). 
Further guidance and information for study intervention preparation and administration and the 
final disposition of unused study interventions are provided in the relevant manuals. 
Details on preparation, handling, storage, and accountability for in-home visits in Part 2 are 
provided in the Home Administration Manual. 
5.4. 
Measures to Minimize Bias: Randomization 
In Part 1, eligible subjects will be randomized in the order they are enrolled into the study. For 
Cohort 1, 8 eligible subjects will be randomized in a 3:1 ratio to 
active (BIO89-100) : control (placebo). For Cohort 2, 15 subjects will be randomized in a 4:1 
ratio to active (BIO89-100) : control (placebo). For Cohorts 4 and 6, 12 eligible subjects will be 
randomized in each cohort in a 3:1 ratio to active (BIO89-100) : control (placebo). For Cohorts 3 
and 5, 18 subjects will be randomized in each cohort in a 7:2 ratio to 
active (BIO89-100) : control (placebo), stratified by biopsy-confirmed NASH with fibrosis status 


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F1, F2 or F3 (Yes, No) (Kleiner, 2005), with each stratum of 9 subjects in each cohort. All 
subjects in Part 1 will be centrally assigned to randomized study intervention using an Interactive 
Response Technology (IRT). 
Part 1 will be conducted under double-blind conditions. The subjects, Principal Investigator, 
other study personnel involved with subject assessments, and the Sponsor will remain blinded to 
the actual treatment assignments of the subjects. Blinded study intervention will be prepared by a 
clinical supplies vendor and shipped to the study site. Where applicable, an unblinded pharmacist 
at the study site will prepare the study intervention for SC administration. 
Part 2 will be open-label with all subjects receiving the same dose of BIO89-100. 
5.5. 
Study Intervention Compliance 
When subjects are dosed at the site, they will receive study intervention directly from the 
Investigator or designee, under medical supervision. The date and time of each dose 
administered in the clinic will be recorded in the source documents and recorded in the electronic 
case report form (eCRF). The dose of study intervention and study subject identification will be 
confirmed at the time of dosing by a member of the study site staff other than the person 
administering the study intervention. 
Details on study intervention compliance at home visits in Part 2 are provided in the Home 
Administration Manual. In cases in which a dose is administered outside of the allowed visit time 
window, adjustments to the remaining visit dates may be required. The investigator should 
discuss with the Sponsor if the dosing schedule has changed markedly (e.g., ≥1 week) compared 
to the original schedule. 
5.6. 
Concomitant Therapy 
Any medication or vaccine (including over-the-counter or prescription medicines, vitamins, and 
herbal supplements) that the subject is receiving at the time of enrollment or receives during the 
study must be recorded along with at least the following information: 
• Reason for use 
• Dates of administration including start and end dates 
• Dosage information including dose and frequency 
It has previously been reported that NAFLD is associated with reduced CYP3A drug metabolism 
activity in vivo and hepatic CYP3A4 expression in humans (Woolsey, 2015). In addition, based 
on the work by Woolsey et al it is suggested that exogenously administered FGF21 may decrease 
hepatic CYP3A4 activity (Woolsey, 2016) . The underlying mechanism has not been fully 
elucidated. 
However, subjects receiving concomitant administration of CYP3A4 substrates with narrow 
therapeutic index should be closely monitored for any potentially related safety events, and dose 
adjustment of the CYP3A4 substrate should be considered as needed.  
The CRO Medical Monitor should be contacted if there are any questions regarding concomitant 
or prior therapy. 


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5.6.1. 
Prohibited Medications/Therapies 
Prohibited therapies include: 
• Bariatric surgery (including gastric banding or sleeve surgery) 
• Medications historically associated with secondary NAFLD (e.g., amiodarone, 
methotrexate, systemic glucocorticoids [>2 weeks of pharmacologic dosing], 
tetracyclines, tamoxifen, estrogens at doses greater than those used for hormone 
replacement, anabolic steroids, valproic acid, and other medications with known 
hepatotoxicity). Short term oral steroid bursts with/without taper as well as topical 
and inhaled steroids are allowed. 
• Vitamin E supplementation should not be initiated, or the dose increased during the 
trial  
• Thiazolidinendiones.  
• Any investigational product, FGF21 analogues (e.g., LY2405319, LY3025876, 
BMS-986036, BMS-986171, PF-05231023, AKR-001, BIO89-100), or FGFR1-
activating products. 
5.7. 
Stopping Rules and Dose Modifications 
The Sponsor may suspend or terminate the study in the event of: 
• New toxicological or pharmacological findings or clinical safety findings that 
invalidate the earlier positive benefit-risk assessment. 
• Discontinuation of the development of BIO89-100. 
• External circumstances that do not enable the study to be properly conducted under 
the existing protocol, including potential circumstances related to the COVID-19 
2020 pandemic. 
In Part 1, if 3 or more subjects within a single cohort meet stopping rules criteria, as described 
below, the SMC will meet to decide on study/dose-level discontinuation, suspension, dose 
modification or repetition prior to proceeding to subsequent dosing: 
• New elevation of liver transaminase (ALT or AST) or bilirubin from baseline, as 
described below, without evidence of an alternative etiology for this elevation 
• Serious hypersensitivity reactions to BIO89-100 
• Any SAE on a case-by-case basis 
• A concerning pattern of laboratory or other safety findings 
5.7.1. 
Monitoring and Discontinuation for Suspected Drug-induced Liver Injury 
(DILI)  
Per FDA recommendations, the following criteria for elevations in liver transaminases or 
bilirubin will be used for closely monitoring, discontinuing or temporarily interrupting study 
intervention. 
Definition of baseline ALT and AST values 


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Baseline value is defined as an average of all ALT and AST values performed during screening 
and the Baseline (Day 1) visit, as follows:  
ALT/AST Screening Assessments 
Day 1 
ALT/AST 
Assessment 
Baseline Value  
Assessment 1 
Assessment 2  
Assessment 3 (if 
applicable) 
Normal 
Normal 
Not applicable 
Any 
Average of 
Assessment 1, 
Assessment 2 and 
Day 1 (3 tests) 
Normal  
Abnormal and 
≤40% increase 
from Assessment 1 
Not applicable 
Any 
Average of 
Assessment 1, 
Assessment 2 and 
Day 1 (3 tests)  
Normal  
Abnormal and 
>40% increase 
from Assessment 1 
Normal or ≤40% 
increase from 
Assessment 1 
Any 
Average of 
Assessment 1, 
Assessment 2, 
Assessment 3 and 
Day 1 (4 tests) 
Abnormal and >40% 
increase from 
Assessment 1 
Not applicable, 
subject 
excluded 
Not applicable, 
subject excluded 
Abnormal  
≤40% increase 
from Assessment 1 
Not applicable 
Any 
Average of 
Assessment 1, 
Assessment 2 and 
Day 1 (3 tests) 
Abnormal 
>40% increase 
from Assessment 1 
≤40% increase from 
Assessment 1 
Any 
Average of 
Assessment 1, 
Assessment 2, 
Assessment 3 and 
Day 1 (4 tests) 
>40% increase from 
Assessment 1 
Not applicable, 
subject 
excluded 
Not applicable, 
subject excluded 
New transaminase elevations in subjects with baseline value within the normal range:  
For new elevations in transaminases to greater than 2x ULN, repeat measurement should be 
performed within 48-72 hours14. If elevations persist, subjects should be evaluated for other 
causes of transaminase elevations and with tests of hepatic function. If no other cause is 
identified, then the subjects need to be monitored closely (see below), and discontinuation of the 
study drug should be considered.  
 
14 In cases of isolated AST elevation to the indicated threshold, with a clear non-hepatic source for AST elevation 
(e.g. evidence of significant concurrent creatine phosphokinase elevation), decision regarding need to proceed with 
DILI work-up will be based on investigator judgement.  


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Drug should be discontinued, and the subject followed until resolution of symptoms or signs in 
the following situations: 
• ALT or AST >8x ULN 
• ALT or AST >5x ULN for more than 2 weeks 
• ALT or AST >3x ULN and (TB >2x ULN or INR >1.5) 
• ALT or AST >3x ULN with the appearance of fatigue, nausea, vomiting, right upper 
quadrant pain or tenderness, fever, rash, and/or eosinophilia (>5%) 
New transaminase elevations in subjects with baseline ALT or AST > ULN  
For new elevations in transaminases to greater than 2x baseline value or total bilirubin > 1.5x 
ULN, repeat measurement should be performed within 48-72 hours15. If elevations persist, 
subjects should be evaluated for other causes of transaminase elevations and with tests of hepatic 
function. If no other cause is identified, then the subjects need to be monitored closely (see 
below), and discontinuation of the study drug should be considered. 
Study intervention should be discontinued, and the subject followed until resolution of symptoms 
or signs in the following situations: 
Table 7 
Discontinuation Criteria in Subjects with Abnormal Baseline ALT or AST 
Values 
Baseline Value of ALT/AST 
Criteria to Discontinue Study Intervention 
<2× upper limit of normal (ULN) 
if ALT or AST increases to >5× baseline value 
≥2× ULN but <5× ULN 
if ALT or AST increases to >3× baseline value 
≥5× ULN 
if ALT or AST increases to >2× baseline value 
Other 
if ALT or AST increase to >2× baseline value AND the increase 
is accompanied by a concomitant total bilirubin increase to >2× 
ULN OR the INR concomitantly increases by >0.2.  
if ALT or AST increase to >2× baseline value in the presence of 
signs and symptom(s) such as fatigue, nausea, vomiting, right 
upper quadrant pain or tenderness, fever, rash, and/or eosinophil 
(>5%) 
ALT = alanine aminotransferase; AST = aspartate aminotransferase; INR = normalized ratio 
Close Monitoring for Suspected DILI: 
• Repeating liver enzyme and serum bilirubin tests two or three times weekly. 
Frequency of repeat testing can decrease to once a week or less if abnormalities 
stabilize or the trial drug has been discontinued and the subject is asymptomatic. 
• Obtaining a more detailed history of symptoms and prior or concurrent diseases. 
• Obtaining a history of concomitant drug use (including nonprescription medications 
and herbal and dietary supplement preparations), alcohol use, recreational drug use, 
and special diets. 
• Rule out acute viral hepatitis types A, B, C, D, and E; autoimmune or alcoholic 
hepatitis; hypoxic/ischemic hepatopathy; and biliary tract disease. 


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• Obtaining a history of exposure to environmental chemical agents. 
• Obtaining additional tests to evaluate liver function, as appropriate (e.g., INR, direct 
bilirubin). 
• Considering gastroenterology or hepatology consultations. 
Note: If a patient lives in a remote area, laboratory testing can be performed locally and the 
results should be promptly communicated to the investigator site. 
5.8. 
Screen Failures 
Screen failures are defined as subjects who consent to participate in the clinical study but are not 
subsequently randomized. A minimal set of screen failure information is required to ensure 
transparent reporting of screen failure subjects 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 study (screen failure) may be 
rescreened at a later date if the reason for screen failure is deemed temporary and is resolved 
prior to re-screening (e.g. respiratory infection). Subjects can only re-screen once. 
5.9. 
Intervention after the End of Study 
No additional intervention is planned beyond the end of the study. 


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6. 
DISCONTINUATION OF STUDY INTERVENTION AND 
SUBJECT DISCONTINUATION/WITHDRAWAL 
6.1. 
Discontinuation of Study Intervention 
In rare instances, it may be necessary for a subject to permanently discontinue study intervention. 
If study intervention is permanently discontinued, the subject will remain in the study to be 
evaluated for safety, tolerability, and PK/PD assessments. See the SoAs (Table 2 for Cohorts 1-4, 
Table 4 for Cohorts 5-6, and Table 6 for Cohort 7) for data to be collected at the time of 
discontinuation of study intervention (ET visit). 
Please also refer to stopping rules described in Section 5.7  
6.2. 
Subject Discontinuation/Withdrawal from the Study 
A subject may withdraw or be withdrawn from the study for the following reasons: 
• Investigator decision 
• AE or intercurrent illness 
• Noncompliance with protocol requirements 
• Subject withdrawal of consent 
• Pregnancy 
• Sponsor termination or suspension of the study 
• Lost to follow-up 
The reason for subject withdrawal from the study will be recorded in the eCRF.  
Pregnancy is a mandatory criterion for permanent discontinuation of study intervention. 
At the time of withdrawal from the study, if possible, subject should attend the ET visit, as 
shown in the SoAs (Table 2 for Cohorts 1-4 , Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). 
6.3. 
Lost to Follow-up 
A subject will be considered lost to follow-up if he or she repeatedly fails to return for scheduled 
visits and is unable to be contacted by the study site. 
The following actions must be taken if a subject fails to return to the clinic for a required study 
visit: 
• The site must attempt to contact the subject and reschedule the missed visit as soon as 
possible and counsel the subject on the importance of maintaining the assigned visit 
schedule and ascertain whether or not the subject wishes to or should continue in the 
study. 
• Before a subject is deemed lost to follow-up, the Investigator or designee must make 
every effort to regain contact with the subject (where possible, 3 telephone calls and, 
if necessary, a certified letter to the subject’s last known mailing address or local 
equivalent methods). These contact attempts should be documented in the subject’s 
medical record. 


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• Should the subject continue to be unreachable, he or she will be considered to have 
withdrawn from the study (and be labelled as lost to follow-up). 
• Discontinuation of specific sites or of the study as a whole are handled as part of 
Appendix 1 (Section 9.1.8). 


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7. 
STUDY ASSESSMENTS AND PROCEDURES 
• Study procedures and their timing are summarized in the SoAs (Table 2 for 
Cohorts 1-4, Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). Protocol waivers or 
exemptions are not allowed. 
• If multiple assessments are scheduled at the same timepoint, it is recommended that 
procedures be performed in the following sequence: 12-lead ECGs, vital signs, 
sample collection for laboratory and PD biomarker tests, and sample collection for 
PK. 
• Immediate safety concerns should be discussed with the Sponsor immediately upon 
occurrence or awareness to determine if the subject should continue or discontinue 
study intervention. 
• Adherence to the study design requirements, including those specified in the SoAs, is 
essential and required for study conduct. 
• Any situation at the site level with potential impact on subject safety or study 
conduct, including situations related to COVID-19 infection or control measures, 
should be discussed with the Sponsor immediately upon occurrence or awareness to 
determine potential impact on study subject/s or study conduct.  
• All screening evaluations must be completed and reviewed to confirm that potential 
subjects meet all eligibility criteria. The Investigator will maintain a screening log to 
record details of all subjects screened and to confirm eligibility or record reasons for 
screening failure, as applicable. 
• Procedures conducted as part of the subject’s routine clinical management (e.g., blood 
count) and obtained before signing of the ICF may be utilized for screening or 
baseline purposes provided the procedures met the protocol-specified criteria and 
were performed within the time frame defined in the SoAs (Table 2 for Cohorts 1-4, 
Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). 
7.1. 
Efficacy Assessments 
The PD and biomarker assessments are described in Section 7.6. 
7.1.1. 
Liver Biopsy and Scoring (Part 2 Only) 
Liver biopsy remains the gold standard for diagnosing NASH as well as for establishing the 
degree of liver fibrosis as it can reliably assess the various histopathological patterns 
characteristic of NASH (Arab, 2018; Bedossa, 2018; Gunn, 2018; Isabela Andronescu, 2018). In 
addition, histopathology scores are used to assess extent of fibrosis before and after treatment. 
The fibrosis stage ranges from F0 (without fibrosis) to F4 (cirrhosis). Currently, liver biopsy is 
required by regulatory authorities as a surrogate endpoint to assess drug efficacy in Phase 3 
clinical studies. 
In Part 2, paired liver-biopsy will be performed percutaneously per institution standard, at 
screening and within 14 days after the last dose of study intervention (as specified in the SoA 
(Table 6). Details on the biopsy procedure, preparation of slides and central read are provided in 


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the Biopsy Manual. The procedure will be performed by an experienced hepatologist or 
radiologist. Biopsies will be subject to central read by at least one liver pathologist. 
In addition to the central read, liver histology will also be quantitatively assessed by an AI-based 
machine-read analysis. Additional details are provided in the Biopsy Manual.  
7.2. 
Safety Assessments 
Safety assessments include AEs (either reported by the subject or observed by the Investigator), 
concomitant medication use, physical examination, ECG, vital signs, and laboratory assessments. 
Planned time points for all safety assessments are provided in the SoAs (Table 2 for Cohorts 1-4, 
Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). 
7.2.1. 
Physical Examinations 
• A complete physical examination will include, at a minimum, assessments of the 
cardiovascular, respiratory, abdominal and a neurological exam. Height (only 
collected at screening) and weight will also be measured and recorded. 
• A symptom directed physical examination will include, at a minimum, assessments of 
the skin, respiratory, cardiovascular system, and abdomen (liver and spleen). 
7.2.2. 
Vital Signs 
• Vital signs include blood pressure, pulse, body temperature, and respiratory rate. 
• Planned time points for vital signs are provided in the SoAs (Table 2 for Cohorts 1-4, 
Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). 
• Starting from randomization, blood pressure and pulse will be measured in duplicate, 
the first measurement will be taken up to 15 minutes before the indicated time point. 
Additional vital signs measurement may be done if clinically indicated. Subjects must 
be in a supine or semi-erect/seated position and resting for at least 5 minutes prior to 
measurements. 
7.2.3. 
Electrocardiograms 
• 12-lead ECG will be recorded as single bedside measurements using an ECG machine 
that automatically calculates the heart rate and measures PR, QRS, and QT (QTcF) 
intervals. 
• Subject to be resting for at least 2 minutes prior to ECG. 
7.2.4. 
Columbia-Suicide Severity Rating Scale (C-SSRS) 
The Columbia Suicide Severity Rating Scale (C-SSRS) is a semi-structured interview that 
assesses suicidal ideation and behaviors (Posner, 2011). The C-SSRS can be administered by a 
medical professional (MD, DO, PhD, PA-C, NP, RN) or research coordinator who has 
documented training of the C-SSRS. 
The “Baseline/Screening Version” C-SSRS will be administered at Day -1 (Part 1) or during 
screening (Part 2) to exclude subjects with any past or current suicidal ideations or attempts. 
Subjects will be excluded from the study if they answer “yes” to any one of the suicidal ideation 


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or suicidal behavior questions. The “Since Last Visit” version will be administered in Part 1 only 
at subsequent visits as specified in the SoAs (Table 2 for Cohorts 1-4, Table 4 for Cohorts 5-6). 
Any clinically significant change from baseline may be recorded as an AE if deemed appropriate 
by the Investigator or Sponsor. 
7.2.5. 
Clinical Safety Laboratory Assessments 
See Appendix 2 (Section 9.2) for the list of clinical laboratory tests to be performed and the 
SoAs (Table 2 for Cohorts 1-4, Table 4 for Cohorts 5-6, and Table 6 for Cohort 7) for the timing 
and frequency. Laboratory tests should be performed under fasting conditions (≥10 hours). 
Fasting includes food and all beverages except for non-mineral water. It is recommended that 
subjects abstain from consumption of alcoholic beverages for at least 24 hours before laboratory 
test assessments. 
The Investigator must review the laboratory report, document this review, and record any 
clinically relevant changes occurring during the study in the AE section of the eCRF. The 
laboratory reports must be filed with the source documents. Clinically significant abnormal 
laboratory findings are those which are not associated with the underlying disease, unless judged 
by the Investigator to be more severe than expected for the subject's condition. 
All laboratory tests with values considered clinically significantly abnormal during participation 
in the study or within 28 days after the last dose of study intervention should be repeated until 
the values return to normal or baseline or are no longer considered clinically significant by the 
Investigator or Sponsor’s 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 2 (Section 9.2), must be 
conducted in accordance with the relevant study manual and the SoAs (Table 2 for Cohorts 1-4, 
Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). 
If laboratory values from non-protocol specified laboratory assessments performed at the 
institution’s local laboratory require a change in subject management or are considered clinically 
significant by the Investigator (e.g., SAE or AE or skipping of dose), then the results must be 
recorded in the eCRF and the Sponsor should be notified. 
7.3. 
Adverse Events and Serious Adverse Events 
The definitions of an AE or SAE can be found in Appendix 3 (Section 9.3). 
AEs will be reported by the subject (or, when appropriate, by a caregiver, surrogate, or the 
subject's legally authorized representative). 
The Investigator and any qualified designees are responsible for detecting, documenting, and 
recording events that meet the definition of an AE or SAE and remain responsible for following 
up AEs that are serious, considered related to the study intervention or study procedures, or that 
caused the subject to discontinue the study or study intervention (see Section 6). 


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7.3.1. 
Time Period and Frequency for Collecting AE and SAE Information 
All AEs and SAEs will be as specified in the SoAs (Table 2 for Cohorts 1-4, Table 4 for 
Cohorts 5-6, and Table 6 for Cohort 7). 
Table 8 below summarizes the different reporting periods for AEs, SAEs, and events to monitor. 
Events to monitor are defined in Section 9.3.3. 
Table 8 
Adverse Event Reporting Periods 
Type of 
Event 
Adverse Event 
Serious Adverse Event 
Events to monitor with 
study intervention 
Reporting 
period 
From consent until 
28 days after last dose of 
study intervention 
From consent until 28 days 
after the last dose of study 
intervention  
From consent until 28 days 
after the last dose of study 
intervention 
Reporting 
Timelines to 
the Sponsor  
Entered into the clinical 
database on an ongoing 
basis   
Within 24 hours  
Within 24 hours 
All SAEs will be recorded and reported to the Sponsor or designee immediately and under no 
circumstance should this exceed 24 hours, as indicated in Appendix 3 (Section 9.3). The 
Investigator will submit any updated SAE data to the Sponsor or designee 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 subject has been discharged from the study, and he/she considers the event to be reasonably 
related to the study intervention or study participation, the Investigator must promptly notify the 
Sponsor or designee. 
7.3.2. 
Method of Detecting AEs and SAEs 
The method of recording, evaluating, and assessing causality of AE and SAE and the procedures 
for completing and transmitting SAE reports are provided in Appendix 3 (Section 9.3). 
Care will be taken not to introduce bias when detecting AEs or SAEs. Open-ended and 
non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrences. 
7.3.3. 
Follow-up of AEs and SAEs 
After the initial AE or SAE report, the Investigator is required to proactively follow each subject 
at subsequent visits/contacts. All SAEs, will be followed until resolution, stabilization, the event 
is otherwise explained, or the subject is lost to follow-up (as defined in Section 6.3). Further 
information on follow-up procedures is given in Appendix 3 (Section 9.3). 
7.3.4. 
Regulatory Reporting Requirements for SAEs 
• Prompt notification by the Investigator to the Sponsor or designee of an SAE is 
essential so that legal obligations and ethical responsibilities towards the safety of 
subjects and the safety of a study intervention under clinical investigation are met. 


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• The Sponsor has a legal responsibility to notify both the local regulatory authority 
and other regulatory agencies about the safety of a study intervention under clinical 
investigation. The Sponsor will comply with country-specific regulatory requirements 
relating to safety reporting to the regulatory authority, Institutional Review Boards 
(IRBs)/Independent Ethics Committees (IECs), and investigators. 
• Investigator safety reports must be prepared for suspected unexpected serious adverse 
reactions (SUSARs) according to local regulatory requirements and Sponsor policy 
and forwarded to investigators as necessary. 
• An Investigator who receives an Investigator safety report describing an 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 IRB/IEC, if appropriate 
according to local requirements. 
7.3.5. 
Pregnancy 
• Details of all pregnancies in female subjects and of female partners of male subjects 
will be collected after the start of study intervention and until 90 days after last dose 
of study intervention. 
• 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 4. 
• Abnormal pregnancy outcomes (e.g., spontaneous abortion, fetal death, stillbirth, 
congenital anomalies, ectopic pregnancy) are considered SAEs. 
7.4. 
Treatment of Overdose 
There is no experience with overdose of BIO89-100 in humans.  
For this study, any dose of BIO89-100 greater than the protocol-specified dose will be 
considered an overdose. 
The Sponsor does not recommend specific treatment for an overdose. 
In the event of an overdose, the Investigator should: 
• Contact the CRO Medical Monitor immediately. 
• Closely monitor the subject for any AEs and laboratory abnormalities until study 
intervention can no longer be detected systemically (at least 30 days). 
• Obtain a plasma sample for PK analysis within 2 days from the date of the overdose if 
requested by the Medical Monitor (determined on a case-by-case basis). 
• Document the quantity of the excess dose as well as the incidence of the overdose in 
the eCRF. 


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7.5. 
Pharmacokinetics 
Blood samples for analysis of BIO89-100 serum levels will be collected at the timepoints 
designated in the SoAs (Table 3 for Cohorts 1-4, Table 5 for Cohorts 5-6, and Table 6 for 
Cohort 7). 
Blood samples will be processed for collection of serum fractions for determination of 
BIO89-100 serum concentrations. Serum samples will be shipped to the bioanalytical laboratory 
for analysis. 
The trough serum BIO89-100 concentrations will be summarized by treatment group and 
nominal sampling time using descriptive statistics. 
Pharmacokinetic parameters calculated by noncompartmental methods from the BIO89-100 
serum concentration data will include: 
PK Parameter 
Cohorts 1-3 
Cohorts 4-5 
Cmax 
Within 1st and 5th dosing intervals 
Within 1st and 3rd dosing intervals 
AUC0-tau 
Within 1st and 5th dosing intervals 
Within 1st and 3rd dosing intervals 
tmax  
Within 1st and 5th dosing intervals 
Within 1st and 3rd dosing intervals 
t½  
Within 1st and 5th dosing intervals 
Within 1st and 3rd dosing intervals 
Additional PK parameters may be calculated if deemed appropriate. 
The serum concentration-time data may also be used for population PK modeling with the results 
reported separately from the clinical study report. 
7.6. 
Pharmacodynamics and Biomarkers 
7.6.1. 
Immunogenicity Assessments 
Antibodies to BIO89-100 as well as titer and binding specificity will be evaluated in serum 
samples collected from all subjects at timepoints designated in the SoAs (Table 2 for 
Cohorts 1-4, Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). Subjects who test positive 
 
 
 
 
 
 More frequent testing (e.g. every month) or testing for a 
longer period of time may be requested in the event of safety-related concerns. Follow-up testing 
will not be required where it is established that the subject did not receive BIO89-100. All 
follow-up results, both positive and negative will be communicated to the sites. A blood sample 
for ADA assessment will also be collected upon observation of any severe hypersensitivity 
reaction (e.g., anaphylaxis). Subjects who test positive 
 
 
 
Samples for endogenous FGF21 level will be collected at baseline for all subjects, and at the 
time of acquisition of the follow-up blood sample(s) for ADA assessment, in 
 


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Serum samples will be screened for antibodies binding to BIO89-100 and the titer of confirmed 
positive samples will be reported. Other analyses may be performed to verify the stability of 
antibodies to BIO89-100 and/or further characterize the immunogenicity of BIO89-100. 
The detection and characterization of antibodies to BIO89-100 will be performed using a 
validated assay method under the supervision of the Sponsor. Antibodies may be further 
characterized and/or evaluated for their ability to neutralize the activity of the study 
intervention(s). Samples may be stored for a maximum of 
 (or according to local 
regulations) following the last subject’s last visit for the study at a facility selected by the 
Sponsor to enable further analysis of immune responses to BIO89-100. 
7.6.2. 
Pharmacodynamics 
The following biomarkers/PD parameters will be evaluated at timepoints designated in the SoAs 
(Table 2 for Cohorts 1-4, Table 4 for Cohorts 5-6, and Table 6 for Cohort 7): 
Laboratory Parameters 
• Triglycerides 
• Non-HDL cholesterol 
• HDL-c 
• LDL-c 
• HbA1c 
• HOMA-IR (Part 1 only) 
• Liver function tests: ALT, AST  
• GGT 
• Alkaline phosphatase 
• Adiponectin  
• Pro-C3 (see Section 7.6.2.4) 
• Free fatty acids and Adipo-IR (fasting free fatty acids  fasting insulin; Part 1 only) 
 
 
 
  
 
 
 
 
 
 
 
 


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Samples may be stored for a maximum of 15 years after the last subject’s last visit for the study, 
at a facility selected by the Sponsor, to enable further analysis of biomarker responses to 
BIO89-100. 
Residual blood samples will be stored for potential future analysis of biomarkers. 
Additional information will be available in the study specific laboratory manual. 
Other parameters include: 
• Imaging measures (Section 7.6.2.5) 
− MRI-PDFF  
 
 
 
 
 
 
 
 
 
 
• Anthropomorphic measurements 
− Body weight 
 
  
 
  
 
 
 
 
 
 
 
 
 
 
 
7.6.2.2. 
Homeostatic Model Assessment for Insulin Resistance (HOMA-IR; Part 1 only) 
HOMA-IR is a method used to quantify insulin resistance based on the following formula(s) 
(Matthews, 1985): 


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Glucose in mmol/L 
 
Glucose in mg/dL 
HOMA-IR = 
Glucose × Insulin 
 
HOMA-IR = 
Glucose × Insulin 
22.5 
 
405 
 
 
 
 
 
 
7.6.2.4. 
N-terminal Propeptide of Type III Collagen (Pro-C3) 
N-protease cleaved N-terminal propeptide of type 3 procollagen (P3NP) neo-epitope (Pro-C3) is 
derived from the synthesis of type 3 collagen. Pro-C3 appears to correlate with liver fibrosis 
stage, fibrosis regression and response to treatment both as a single test and as part of algorithms 
(Nielsen, 2015; Daniels, 2018; Hansen, 2018). 
7.6.2.5. 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction 
(MRI-PDFF)  
MRI-PDFF is as a noninvasive, quantitative, and accurate measure of liver fat content (imaging-
based biomarker) to assess treatment response in NASH clinical studies (Caussy, 2018). 
This technology enables post-processing of MRI data into parametric map of PDFF (Antaros 
Medical, Sweden) to provide accurate and quantitative measures of liver fat.  
 
 
 
 
 
 
 
 
 
PDFF 
The PDFF is determined using a 6 echo gradient echo pulse sequence covering the liver in the 
axial plane. Analysis is performed by semi-automatic contouring of the liver in every slice 
avoiding major vessels and bile ducts. The method applies multi-peak lipid spectral models and 
simultaneous quantification and correction for T2*. The liver fat value (PDFF) is the mean value 
of all voxels in the identified volume of interest.  
For more information, refer to the Imaging Manual. 


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7.6.3. 
Exploratory Analyses 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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8. 
STATISTICAL CONSIDERATIONS 
This section describes the statistical analysis as foreseen at the time of planning the study. 
Changes, additions, and further details about the analyses will be described in the statistical 
analysis plan, if applicable. Any subsequent additional analyses or changes to analyses that may 
be required will be fully disclosed in the clinical study report. Summaries and analyses will be 
presented by dose/treatment group for Part 1 and separately for Part 2. Before database lock, 
final statistical and pharmacokinetic analysis plans containing full details of all planned analyses 
will be produced. The analyses presented here represent an outline of the intended methodology. 
Details of the analysis will be described in a Statistical Analysis Plan. All clinical data will be 
summarized using descriptive statistics: number (n), mean, standard deviation (SD), median, 
minimum, and maximum for continuous measurements, and counts and percentages for 
categorical measurements. 
8.1. 
Statistical Hypotheses 
The Part 1 primary objectives are to evaluate the safety and tolerability of ascending multiple SC 
injections of BIO89-100 in subjects with NASH or who have NAFLD and at a high risk of 
NASH and to characterize BIO89-100 PK.  No formal hypotheses will be tested for the primary 
objectives. 
The secondary objectives of evaluating the effect of BIO89-100 on selected PD markers and 
biomarkers will be assessed by point estimates, 90% confidence intervals and the descriptive 
p-values of mean changes/percentages changes from baseline within and between treatment 
group. 
For Part 2, the primary objectives are to evaluate incidence of AEs and to assess histological 
response of BIO89-100 in the NASH population. 
8.2. 
Sample Size Determination 
For Part 1, no formal sample size calculation was performed for the primary endpoint(s) as the 
total of approximately 83 subjects, consisting of 6:2 (Cohort 1), 12:3 (Cohort 2), 9:3 (Cohorts 4 
and 6), or 14:4 (Cohorts 3 and 5) receiving active : placebo, respectively is considered adequate 
for Part 1 of this study and are expected to enroll to achieve study objectives. 
A power assessment based on 
 
 subjects in a dose 
cohort compared to the pooled 19 placebo subjects will provide 
 power, 
respectively, to detect a difference in mean percentage change from baseline between treatment 
groups of 
, assuming the standard deviation of endpoint in each group is 
. Calculation 
was based on two-sample t-test with one-sided 5% (e.g., two-sided 10%) type I error probability. 
Subjects who withdraw from the study before the Day 50 assessments may be replaced at the 
discretion of the Sponsor. 
For Part 2, no formal power calculations were used to determine the sample size in Cohort 7 and 
the number of subjects was chosen based on clinical experience of similar proof-of-concept 
histology studies and feasibility. This sample size accounts for an anticipated 
 of subjects 
who will not undergo the post-baseline liver biopsy (e.g., 
 to have paired liver biopsies). 
With a sample size of 
, the 95% confidence interval of the histological response rate would 


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have a half-width no greater than 
 assuming the histological response 
 The 
actual half-width will depend on the observed histological response rate. 
8.3. 
Populations for Analyses 
For purposes of analysis, the following populations are defined: 
Population 
Description 
Screened Analysis set 
All enrolled subjects who signed informed consent and undergone screening. 
Part 1 
 
Randomized Analysis 
set 
All enrolled subjects who are assigned a randomization number in the study. 
For analysis purposes, subjects will be analyzed according to the treatment 
they were randomized to regardless of actual treatment received. 
Safety Analysis set 
All randomized subjects who receive at least 1 of study intervention. In this 
population, subjects will be summarized based upon the study intervention 
actually received, regardless of the study intervention to which they were 
randomized. 
PK Analysis set 
All subjects in the Safety Analysis set who have sufficient data to adequately 
characterize the trough serum BIO89-100 concentrations and have no other 
events, or protocol violations that would adversely affect results, such as not 
completing the full dose. The analysis population for any population pk 
modeling may be defined separately in a population PK data analysis plan. 
PD Analysis set 
All subjects in the Safety Analysis set who have measurable post-baseline PD 
data; subjects will summarized based on the randomized treatment group. 
Different PD analysis sets may be used for different types of PD endpoints, 
e.g., MRI-PDFF. 
Part 2 
 
Full Analysis Set 
All enrolled subjects who received at least one dose of study intervention.  
Full Analysis Set will also be used for safety summary in Part 2 
PK Analysis Set 
All enrolled subjects in the Full Analysis Set who have sufficient data to 
adequately characterize the trough serum BIO89-100 concentrations and have 
no other events, or protocol violations that would adversely affect results, such 
as not completing the full dose. 
PD Analysis Set 
All enrolled subjects in the Full Analysis Set who have measurable post-
baseline PD data.  Different PD analysis sets may be used for different types 
of PD endpoints, e.g., MRI-PDFF. 
Biopsy Analysis Set 
For Cohort 7 only. All enrolled subjects in the Full Analysis Set and have both 
baseline and post-baseline biopsies.  
8.4. 
Statistical Analyses 
The statistical analysis plan (SAP) will be developed and finalized before database lock and will 
describe the subject populations to be included in the analyses, and procedures for accounting for 
missing, unused, and spurious data. Pooling of placebo subjects in Part 1 will be performed when 
applicable and will be specified in the analysis plan. This section is a summary of the planned 


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statistical analyses of the primary and secondary endpoints. Analysis of biomarkers and PD 
endpoints as well as their correlation with PK will be detailed in the SAP. 
In general, summary descriptive statistics will be presented for demographics and baseline 
characteristics, safety endpoints, PK, and PD parameters. Descriptive statistics will include 
number of observations, mean, standard deviation, median, minimum, and maximum for 
continuous parameters. For PK and PD parameters, descriptive statistics will include the number 
of observations, arithmetic and geometric means, standard deviation, coefficient of variation, 
median, minimum, and maximum. Categorical parameters will be displayed using counts and 
percentages within each category. 
For Part 1, while no formal hypothesis testing is planned for the primary objectives, the 
evaluation of potential efficacy will be determined based on the totality and pattern of data from 
the biomarker and PD endpoints. The assessment of biomarker and PD endpoints will employ 
90% confidence intervals and significance testing for assessing within group changes and 
percentage changes from baseline and comparisons of the BIO89-100 doses to the pooled 
placebo group. Details will be provided in the SAP. 
The correlation between relevant PK parameters and PD endpoints will be examined as will be 
detailed in the SAP. 
Subjects with dose interruptions impacted by the COVID-19 pandemic will be noted in the 
listings. Subgroups including and excluding subjects with dose interruptions due to COVID-19 
pandemic may be considered for both efficacy and safety analyses as appropriate. 
For Part 2, endpoints of different histological responses will be summarized with the point 
estimates and 95% confidence intervals of the proportions of subjects who meet the response 
criteria using the biopsy analysis set. 
8.4.1. 
Safety and Tolerability Analyses 
All safety analyses will be performed on the Safety Analysis Set in Part 1 and Full Analysis Set 
in Part 2. The safety assessment will be based on AEs, laboratory values, concomitant 
medication use, ECG, vital signs, and physical examination. Summaries will be presented by 
treatment/dose. 
All adverse events will be coded using the Medical Dictionary for Regulatory Activities (current 
version of MedDRA). The severity of adverse events will be graded according to the current 
version of National Cancer Institute (NCI) common terminology criteria for adverse events 
(CTCAE) v5. All AE summaries will include treatment-emergent adverse events (TEAEs), 
defined as those which occur after the first dose of study intervention (BIO89-100 or placebo). 
Treatment-related AEs are those which were determined by the Investigator to be related to study 
intervention. The incidence of treatment-related AEs (as recorded in the eCRF) will be 
summarized using descriptive statistics by system organ class (SOC) and preferred term, 
categorized by severity as recorded in the eCRF. Counting will be done for each subject (subjects 
will only be counted once within each SOC or preferred term). The frequency of TEAE (AE 
burden) will be summarized using descriptive statistics by SOC and preferred term (PT). 
Counting will be done for events (regardless of subjects), by SOC and PT. The incidence of 
TEAE will also be presented by time of onset post-dose, SOC, PT, and severity. 


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A listing of TEAEs and TEAEs leading to study discontinuation will be provided. Summaries 
will be presented by dose/treatment group. Subject listings of SAEs and AE leading to 
withdrawal will be presented. Actual values and changes in laboratory (serum chemistry, 
hematology, coagulation, and urinalysis), ECG, and vital signs measurements including weight 
data will be summarized descriptively. Laboratory data will also be presented by grading 
(severity) scale according to the current version of NCI CTCAE for each time point recorded. All 
values will be compared with prespecified boundaries to identify potentially clinically significant 
values or changes, as detailed in the statistical analysis plan, and such values will be listed. All 
prior and concomitant medications will be coded using the World Health Organization drug 
dictionary (WHO Drug) and will be listed. Concomitant medications will include all medications 
taken from first dose through to the EOS visit. If any subject dies during the study, a listing of 
deaths will be provided and all relevant information will be discussed in the subject narrative 
included in the clinical study report. The relationship of overall safety with respect to 
pharmacokinetics, immunogenicity assessments, and prespecified exploratory biomarkers will be 
evaluated, as appropriate. 
The number (%) of subjects who fail to complete the study and the reason for discontinuation, 
and the number (%) of subjects who fail to complete the study due to related adverse events will 
be summarized. 
8.4.2. 
PK Analyses 
All PK population subjects will be included in the PK analysis. 
Pharmacokinetic parameters including but not limited to those listed in Section 7.5 will be 
determined where possible from the serum concentrations of BIO89-100. 
All PK parameters will be listed by subject and summarized by dose using descriptive statistics 
(sample size, mean, median, geometric mean [as appropriate], coefficient of variation [CV%], 
SD, minimum, and maximum) for continuous variables and using frequencies and percentages 
for discrete variables. 
Serum concentration data will be listed by subject and summarized using descriptive statistics by 
nominal time point. 
Serum concentration by time profiles for each subject will be presented graphically. Mean serum 
concentration by time profiles will be presented by dose level. Further details on PK analysis will 
be provided in the SAP. 
8.4.3. 
Immunogenicity Analyses 
Results of immunogenicity assessment will be provided by ADA response (e.g., number and 
percent of positive ADA subjects) and immunogenicity profile (e.g., antibody titers, binding 
specificity, and neutralizing immunogenicity), when appropriate. 
The effect of ADA on relevant PK parameters may be evaluated. 
For endogenous FGF21, baseline samples will be analyzed; sample(s) will also be obtained from 
 subject with additional follow-up visit(s) for potential future analysis to 
evaluate effect of ADA on endogenous FGF21 level. 


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8.5. 
Interim Analyses 
No efficacy interim analysis is planned. The Sponsor may perform administrative interim 
analysis to support objectives such as study planning and regulatory interaction. 
8.6. 
Safety Monitoring Committee (SMC) 
The SMC, a specific committee for the study composed of the Sponsor’s Medical Monitor, the 
CRO Medical Monitor, and at least one participating Investigator, will only function in Part 1, as 
described in Section 3.2. The operations of the SMC will be defined in the SMC Charter. 
Given the open-label design of Part 2, safety will be reviewed on an ongoing basis by the 
Sponsor and/or CRO Medical Monitors. 


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9. 
SUPPORTING DOCUMENTATION AND OPERATIONAL 
CONSIDERATIONS 
9.1. 
Appendix 1: Regulatory, Ethical, and Study Oversight 
Considerations 
9.1.1. 
Regulatory and Ethical Considerations 
• This study will be conducted in accordance with the protocol and with the following: 
− Consensus ethical principles derived from international guidelines including the 
Declaration of Helsinki and Council for International Organizations of Medical 
Sciences (CIOMS) International Ethical Guidelines 
− Applicable ICH Good Clinical Practice (GCP) Guidelines 
− Applicable laws and regulations 
• The protocol, protocol amendments, ICF, Investigator’s Brochure, and other relevant 
documents (e.g., advertisements) must be submitted to an IRB/IEC by the 
Investigator and reviewed and approved by the IRB/IEC before the study is initiated. 
• Any amendments to the protocol will require IRB/IEC approval before 
implementation of changes made to the study design, except for changes necessary to 
eliminate an immediate hazard to study subjects. 
• The Investigator will be responsible for the following: 
− Providing written summaries of the status of the study to the IRB/IEC annually or 
more frequently in accordance with the requirements, policies, and procedures 
established by the IRB/IEC 
− Notifying the IRB/IEC of SAEs or other significant safety findings as required by 
IRB/IEC procedures 
− Providing oversight of the conduct of the study at the site and adherence to 
requirements of 21 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 
9.1.2. 
Financial Disclosure 
Investigators and sub-investigators will provide the Sponsor with sufficient, accurate financial 
information as requested to allow the Sponsor to submit complete and accurate financial 
certification or disclosure statements to the appropriate regulatory authorities. Investigators are 
responsible for providing information on financial interests during the course of the study and for 
1 year after completion of the study. 
9.1.3. 
Informed Consent Process 
• The Investigator or his or her representative will explain the nature of the study to the 
subject or his or her legally authorized representative and answer all questions 
regarding the study. 


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• Subjects must be informed that their participation is voluntary. Subjects will be 
required to sign a statement of informed consent that meets the requirements of 
21 CFR 50, local regulations, ICH guidelines, Health Insurance Portability and 
Accountability Act (HIPAA) requirements, where applicable, and the IRB/IEC or 
study center. 
• The medical record must include a statement that written informed consent was 
obtained before the subject 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. 
• Subjects must be re-consented to the most current version of the ICF(s) during their 
participation in the study. 
• A copy of the ICF(s) must be provided to the subject or the subject’s legally 
authorized representative. 
Subjects who are rescreened are required to sign a new ICF. 
9.1.4. 
Data Protection 
• Subjects will be assigned a unique identifier by the Sponsor. Any subject records or 
datasets that are transferred to the Sponsor will contain the identifier only; subject 
names or any information which would make the subject identifiable will not be 
transferred. 
• The subject must be informed that his or her personal study-related data will be used 
by the Sponsor in accordance with local data protection law. The level of disclosure 
must also be explained to the subject. 
• The subject must be informed that his/her medical records may be examined by 
Clinical Quality Assurance auditors or other authorized personnel appointed by the 
Sponsor, by appropriate IRB/IEC members, and by inspectors from regulatory 
authorities. 
9.1.5. 
Dissemination of Clinical Study Data 
A clinical study report will be developed by the Sponsor at completion of data analysis. This 
report will be a clinical and statistical integrated report, according to the ICH E3 guidelines. 
Sponsor will register the study and post study results regardless of outcome on a publicly 
accessible website in accordance with the applicable laws and regulations. 
9.1.6. 
Data Quality Assurance 
• All subject data relating to the study will be recorded on the eCRFs unless transmitted 
to the Sponsor or designee electronically (e.g., laboratory data). The Investigator is 
responsible for verifying that data entries are accurate and correct by electronically 
signing the eCRF. 
• The Investigator must maintain accurate documentation (source data) that supports 
the information entered in the eCRF. 


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• The Investigator must permit study-related monitoring, audits, IRB/IEC review, and 
regulatory agency inspections and provide direct access to source data documents. 
• Monitoring details describing strategy (e.g., risk-based initiatives in operations and 
quality such as Risk Management and Mitigation Strategies and Analytical Risk-
Based Monitoring), methods, responsibilities and requirements, including handling of 
noncompliance issues and monitoring techniques (central, remote, or on-site 
monitoring) are provided in the Monitoring Plan. 
• 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 eCRF by authorized site personnel are accurate, complete, and 
verifiable from source documents; that the safety and rights of subjects are being 
protected; and that the study is being conducted in accordance with the currently 
approved protocol and any other study agreements, ICH GCP, and all applicable 
regulatory requirements. It may become required that some or all monitoring 
activities will be halted or performed remotely due to the recent COVID-19 
pandemic. In such a case, details of the changes made to the monitoring strategy will 
be described in the Monitoring Plan.  
• Records and documents, including signed ICFs, pertaining to the conduct of this 
study must be retained by the Investigator per ICH-GCP and local regulations or 
institutional policies. No records may be destroyed during the retention period 
without the written approval of the Sponsor. No records may be transferred to another 
location or party without written notification to the Sponsor, whether within the 
retention period or thereafter. 
9.1.7. 
Source Documents 
• Source documents provide evidence for the existence of the subject and substantiate 
the integrity of the data collected. Source documents are filed at the Investigator’s 
site. 
• Data entered in the eCRF that are transcribed from source documents must be 
consistent with the source documents or the discrepancies must be explained. The 
Investigator may need to request previous medical records or transfer records, 
depending on the study. Also, current medical records must be available. 
• Source documents are original documents, data, and records from which the subject’s 
eCRF data are obtained. These include but are not limited to hospital records, clinical 
and office charts, laboratory and pharmacy records, diaries, microfiches, radiographs, 
and correspondence. 
9.1.8. 
Study and Site Closure 
The Sponsor designee reserves the right to close the study site or terminate the study at any time 
for any reason at the sole discretion of the Sponsor. Study sites will be closed upon study 


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completion. A study site is considered closed when all required documents and study supplies 
have been collected and a study-site closure visit has been performed. 
The Investigator may initiate study-site closure at any time, provided there is reasonable cause 
and sufficient notice is given to the Sponsor in advance of the intended termination. 
Reasons for the early closure of a study site by the Sponsor or Investigator may include but are 
not limited to: 
• Failure of the Investigator to comply with the protocol, the requirements of the 
IRB/IEC or local health authorities, the Sponsor's procedures, or GCP guidelines 
• Inadequate recruitment of subjects by the Investigator 
• Discontinuation of further study intervention development 
If the study is prematurely terminated or suspended, the Sponsor shall promptly inform the 
Investigators, the IECs/IRBs, the regulatory authorities, and any contract research 
organization(s) used in the study of the reason for termination or suspension, as specified by the 
applicable regulatory requirements. The Investigator shall promptly inform the subject and 
should assure appropriate subject therapy and/or follow-up. 
9.1.9. 
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 the International 
Committee of Medical Journal Editors authorship requirements. 
 
 


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9.2. 
Appendix 2: Clinical Laboratory Tests 
• The clinical laboratory tests detailed in Table 9 will be performed by a central 
laboratory, except if noted otherwise, at timing/frequency detailed in the SoAs 
(Table 2 for Cohorts 1-4, Table 4 for Cohorts 5-6, and Table 6 for Cohort 7). 
Laboratory tests will be performed under fasting conditions (≥10 hours). 
• Protocol-specific requirements for inclusion or exclusion of subjects are detailed in 
Section 3.3 and Section 4.3 of the protocol for Parts 1 and 2, respectively.  
• Additional tests may be performed at any time during the study as determined 
necessary by the Investigator or required by local regulations. 
Table 9 
Protocol-Required Laboratory Assessments 
Hematology 
 
White blood cell count (WBC) with differential 
(Neutrophils, Lymphocytes, Monocytes, 
Eosinophils, Basophils – absolute and %) 
Red blood cell (RBC) 
RBC Indices: 
MCV, MCH, %Reticulocytes 
Hemoglobin 
Hematocrit 
Platelet count 
Coagulation factors, including PT, INR, aPTT  
Red cell Distribution Width 
Clinical Chemistries 
 
Alanine Aminotransferase (ALT) 
Aspartate Aminotransferase (AST) 
Alkaline phosphatase (ALP) 
Gamma-glutamyl transferase (GGT) 
Total bilirubin, Indirect/direct bilirubin 
Albumin 
Calcium 
Blood urea nitrogen (BUN) 
Sodium 
Creatinine 
Chloride 
Creatine Kinase 
Magnesium 
Bicarbonate 
Potassium 
Lactate dehydrogenase 
Phosphorus/ 
Total protein 
Glucose (fasting) 
Total Cholesterol 
Triglycerides 
Uric Acid 
High-density lipoprotein cholesterol (HDL-c) 
Low density lipoprotein (LDL-c) 
Non-high-density lipoprotein cholesterol (non-
HDL-c) 


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Urinalysis 
 
Basic Urinalysis (dipstick, including macroscopic 
appearance, bilirubin, blood, color, glucose, 
ketones, leukocyte esterase, nitrite, pH, protein, 
specific gravity, urobilinogen;  
Full urinalysis (dipstick plus microscopic 
evaluation) to be performed only at the Screening 
and End of Study visits). A reflex microscopic 
urinalysis should be performed if the results of the 
urinalysis is abnormal or at the discretion of the 
PI or delegate. 
Other Study-Specific Laboratory Assessments 
 
BIO89-100 (to be evaluated by bioanalytical 
laboratory) 
Insulina 
High-sensitivity C-reactive protein (hsCRP)a 
Oral glucose tolerance test (OGTT) including 
C-peptide, glucose, and insulina 
Hemoglobin A1c (HbA1c) 
Homeostatic model assessment of insulin 
resistance (HOMA-IR)a 
Insulin-like growth factor-1 (IGF-1), totala 
Adiponectin, total 
Cytokeratin (CK)-18a 
Enhanced liver fibrosis (ELF) panela 
N-terminal propeptide of type III collagen 
(Pro-C3) 
Fasting free fatty acids and Adipo-IR (fasting 
FFA x fasting insulin)a 
 
Serum and urine human chorionic gonadotropin 
(hCG) pregnancy test  
TSH 
FSH (only for determination of postmenopausal 
status) 
Urine drug screen including amphetamines, 
barbiturates, cocaine metabolites, opiates, 
benzodiazepines and cannabinoids (on Day -1, can 
done at local laboratory, but must also collect 
sample for central laboratory). 
Virology - HIV antibody, hepatitis B surface 
antigen (HBsAg, and hepatitis C virus antibody) 
24-hour urine for cortisola 
Immunogenicity: 
Antibody to BIO89-100 
Endogenous FGF21 
 
Bone markers - carboxy-terminal collagen 
crosslinks (CTX), N-terminal propeptide of type 1 
collagen (P1NP), osteocalcin  
Plasma and serum samples for exploratory 
biomarkersa 
DNA and RNA samplesa 
a-Part 1 only; all other tests are done in both Parts 1 and 2 
Investigators must document their review of each laboratory safety report. 
 


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9.3. 
Appendix 3: Adverse Events: Definitions and Procedures for 
Recording, Evaluating, Follow-up, and Reporting 
9.3.1. 
Definition of AE 
AE Definition 
• 
An AE is any untoward medical occurrence in a clinical study subject, temporally associated with 
the use of study intervention, whether or not considered related to the study intervention. 
• 
NOTE: An AE can, therefore, be any unfavorable and unintended sign (including an abnormal 
laboratory finding), symptom, or disease (new or exacerbated) temporally associated with the use 
of study intervention. 
 
Events Meeting the AE Definition  
• 
Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) or other safety 
assessments (e.g., ECG, radiological scans, vital signs measurements), including those that worsen 
from baseline, considered clinically significant in the medical and scientific judgment of the 
Investigator (i.e., not related to progression of underlying disease). 
• 
Exacerbation of a chronic or intermittent pre-existing condition including either an increase in 
frequency and/or intensity of the condition. 
• 
New conditions detected or diagnosed after study intervention administration even though it may 
have been present before the start of the study. 
• 
Signs, symptoms, or the clinical sequelae of a suspected drug-drug interaction. 
• 
Signs, symptoms, or the clinical sequelae of a suspected overdose of either study intervention or a 
concomitant medication. Overdose per se will not be reported as an AE/SAE unless it is an 
intentional overdose taken with possible suicidal/self-harming intent. Such overdoses should be 
reported regardless of sequelae. 
 
Events NOT Meeting the AE Definition  
• 
Any clinically significant abnormal laboratory findings or other abnormal safety assessments which 
are associated with the underlying disease, unless judged by the Investigator to be more severe than 
expected for the subject’s condition. 
• 
The disease/disorder being studied or expected progression, signs, or symptoms of the 
disease/disorder being studied, unless more severe than expected for the subject’s condition. 
• 
Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that leads to the 
procedure is the AE. 
• 
Situations in which an untoward medical occurrence did not occur (social and/or convenience 
admission to a hospital). 
• 
Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present or detected at 
the start of the study that do not worsen. 


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9.3.2. 
Definition of Suspected and Unsuspected Adverse Reaction 
Suspected adverse reactions are defined as: 
• 
Any AE for which there is a reasonable possibility that the study intervention caused the AE. For 
the purposes of sponsor regulatory safety reporting, “reasonable possibility” means there is 
evidence to suggest a causal relationship between the study intervention and the AE. A suspected 
adverse reaction implies a lesser degree of certainty about causality than adverse reaction, which 
means any AE caused by a study intervention 
Unexpected AEs are defined as: 
• 
AE which is not listed in the IB of the study intervention or is not listed at the specificity or 
severity that has been observed 
9.3.3. 
Definition of Events to Monitor 
Sponsor-defined Events to Monitor for BIO89-100: 
Events to Monitor include hypersensitivity reactions occurring after the first administration of study 
agent(s) in subjects participating in this clinical study. These events must be reported by the 
Investigator to the sponsor within 24 hours and are to be considered serious (for regulatory reporting 
purposes) only if they meet the definition of an SAE. The events are to be reported on an SAE form  
9.3.4. 
Definition of SAE 
If an event is not an AE per definition in Section 9.3.1, then it cannot be an SAE even if serious 
conditions are met (e.g., hospitalization for signs/symptoms of the disease under study, death due 
to progression of disease). 
An SAE is defined as any untoward medical occurrence that, at any dose: 
1. Results in death 
2. Is life-threatening 
The term 'life-threatening' in the definition of 'serious' refers to an event in which the subject was at 
risk of death at the time of the event. It does not refer to an event, which hypothetically might have 
caused death, if it were more severe. 
3. Requires inpatient hospitalization or prolongation of existing hospitalization 
In general, hospitalization signifies that the subject has been detained (usually involving at least an 
overnight stay) at the hospital or emergency ward for observation and/or treatment that would not have 
been appropriate in the physician’s 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. 
4. 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 


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An SAE is defined as any untoward medical occurrence that, at any dose: 
(e.g., sprained ankle) which may interfere with or prevent everyday life functions but do not 
constitute a substantial disruption. 
5. Is a congenital anomaly/birth defect 
6. Other situations: 
• 
Medical or scientific judgment should be exercised in deciding whether SAE reporting is 
appropriate in other situations such as important medical events that may not be immediately life-
threatening or result in death or hospitalization but may jeopardize the subject or may require 
medical or surgical intervention to prevent one of the other outcomes listed in the above 
definition. These events should usually be considered serious. 
• 
Examples of such events include invasive or malignant cancers, intensive treatment in an 
emergency room or at home for allergic bronchospasm, blood dyscrasias or convulsions that do 
not result in hospitalization, or development of drug dependency or drug abuse. 
9.3.5. 
Recording and Follow-Up of AE and/or SAE 
AE and SAE Recording 
• 
When an AE/SAE occurs, it is the responsibility of the Investigator to review all documentation 
(e.g., hospital progress notes, laboratory reports, and diagnostics reports) related to the event. 
• 
The Investigator will then record all relevant AE/SAE information in the eCRF. 
• 
It is not acceptable for the Investigator to send photocopies of the subject’s medical records to the 
Medical Monitor in lieu of completion of the AE/SAE eCRF page. 
• 
There may be instances when copies of medical records for certain cases are requested by the 
Medical Monitor. In this case, all subject identifiers, with the exception of the subject number, 
will be redacted on the copies of the medical records before submission to the Medical Monitor. 
• 
The Investigator will attempt to establish a diagnosis of the event based on signs, symptoms, 
and/or other clinical information. Whenever possible, the diagnosis (not the individual 
signs/symptoms) will be documented as the AE/SAE. 
Assessment of Intensity 
The severity of each AE will be assessed at onset by a nurse and/or physician. When recording the 
outcome of the AE the maximum severity of the AE experienced will also be recorded. The severity of 
the AE will be graded according to the CTCAE v5.0: 
Grade 1: Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; intervention 
not indicated. 
Grade 2: Moderate; minimal, local, or noninvasive intervention indicated; limiting age-appropriate 
instrumental Activities of Daily Living (ADL)*. 
Grade 3: Severe or medically significant but not immediately life-threatening; hospitalization or 
prolongation of hospitalization indicated; disabling; limiting self-care ADL**. 
Grade 4: Life-threatening consequences; urgent intervention indicated. 
Grade 5: Death related to AE. 
ADL: 
*Instrumental ADL refer to preparing meals, shopping for groceries or clothes, using the telephone, managing 
money, etc. 
**Self-care ADL refer to bathing, dressing and undressing, feeding self, using the toilet, taking medications, and 
not bedridden. 


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AE and SAE Recording 
Assessment of Causality 
The Investigator is obligated to assess the relationship between study intervention and each occurrence 
of each AE/SAE. 
• 
A “"reasonable possibility”" of a relationship conveys that there are facts, evidence, and/or 
arguments to suggest a causal relationship, rather than a relationship cannot be ruled out. 
• 
The Investigator will use clinical judgment to determine the relationship. 
• 
Alternative causes, such as underlying disease(s), concomitant therapy, and other risk factors, 
as well as the temporal relationship of the event to study intervention administration will be 
considered and investigated. 
• 
The Investigator will also consult the Investigator’s Brochure (IB) and/or Product Information, 
for marketed products, in his/her assessment. 
• 
For each AE/SAE, the Investigator must document in the medical notes that he/she has 
reviewed the AE/SAE and has provided an assessment of causality. 
• 
There may be situations in which an SAE has occurred and the Investigator has minimal 
information to include in the initial report to the Medical Monitor. However, it is very 
important that the Investigator always make an assessment of causality for every event 
before the initial transmission of the SAE data to the Medical Monitor. 
• 
The Investigator may change his/her opinion of causality in light of follow-up information and 
send an SAE follow-up report with the updated causality assessment. 
• 
The causality assessment is one of the criteria used when determining regulatory reporting 
requirements. 
Causality Categories: 
• 
UNRELATED - This relationship suggests that there is no association between the study drug 
and the reported event. 
• 
UNLIKELY RELATED - The relationship suggests that the event is more likely due to other 
causes than study drug, however the relationship to the study drug cannot be ruled out. 
• 
POSSIBLY RELATED - This relationship is based on evidence suggesting a causal 
relationship between the study drug and the AE, i.e., there is a reasonable possibility that the 
drug caused the event. The event follows a reasonable temporal sequence from the time of drug 
administration or follows a known response pattern to the study drug but could also have been 
produced by other factors. 
• 
PROBABLY RELATED - This relationship suggests that a reasonable temporal sequence of 
the event with drug administration exists and, based upon the known pharmacological action of 
the drug, known or previously reported adverse reactions to the drug or class of drugs, or 
judgment based on the Investigator’s clinical experience, the association of the event with the 
study drug seems likely. 
• 
DEFINITELY RELATED - This relationship suggests that a definite causal relationship exists 
between drug administration and the AE, and other conditions (concurrent illness, 
progression/expression of disease state, or concurrent medication reaction) do not appear to 
explain the event. 
 


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Follow-up of AEs and SAEs 
• 
The Investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as medically indicated or as requested by the Medical Monitor 
to elucidate the nature and/or causality of the AE or SAE as fully as possible. This may include 
additional laboratory tests or investigations, histopathological examinations, or consultation with 
other health care professionals. 
• 
If a subject dies during participation in the study or during a recognized follow-up period, the 
Investigator will provide the Medical Monitor with a copy of any post-mortem findings including 
histopathology. 
• 
New or updated information will be recorded in the originally completed eCRF. 
• 
The Investigator will submit any updated SAE data to the Sponsor within 24 hours of receipt of 
the information. 
9.3.6. 
Reporting of SAEs 
Reporting of SAEs will be done using eCRF (with paper methods as a back-up in case of system 
unavailability). 
SAE Reporting to the Medical Monitor via an Electronic Data Collection (eCRF) Tool 
• 
The primary mechanism for reporting an SAE to the Medical Monitor will be the electronic data 
collection tool. 
• 
If the electronic system is unavailable, then the site will use the paper SAE data collection tool 
(see next section) in order to report the event within 24 hours.). 
• 
The site will enter the SAE data into the electronic system as soon as it becomes available. 
• 
After the study is completed at a given site, the electronic data collection tool will be taken off-
line to prevent the entry of new data or changes to existing data. 
• 
If a site receives a report of a new SAE from a study subject or receives updated data on a 
previously reported SAE after the electronic data collection tool has been taken off-line, then the 
site can report this information on a paper SAE form (see next section) or to the Medical Monitor 
by telephone. 
• 
Contacts for SAE reporting can be found in the relevant manual. 
 
SAE Reporting to the Medical Monitor via Paper CRF 
• 
Facsimile or email transmission of the SAE paper CRF is the preferred method to transmit this 
information to the Medical Monitor, 
• 
In rare circumstances and in the absence of facsimile or email equipment, notification by 
telephone is acceptable with a copy of the SAE data collection tool sent by overnight mail or 
courier service. 
• 
Initial notification via telephone does not replace the need for the Investigator to complete and 
sign the SAE CRF pages within the designated reporting time frames. 
• 
Contacts for SAE reporting can be found in the relevant manual. 


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9.4. 
Appendix 4: Contraceptive Guidance and Collection of Pregnancy 
Information 
Definitions: 
Woman of Childbearing Potential (WOCBP) 
A woman is considered fertile following menarche and until becoming post-menopausal unless 
permanently sterile (see below). 
If fertility is unclear (e.g., amenorrhea in adolescents or athletes) and a menstrual cycle cannot be 
confirmed before first dose of study intervention, additional evaluation should be considered. 
Women in the following categories are not considered WOCBP 
1. Premenarchal 
2. Premenopausal female with one of the following: 
− hysterectomy 
− bilateral salpingectomy 
− bilateral oophorectomy 
• For individuals with permanent infertility due to an alternate medical cause other than 
the above, (e.g., Mullerian agenesis, androgen insensitivity), Investigator discretion 
should be applied to determining study entry. 
3. Postmenopausal female 
• A postmenopausal state is defined as no menses for 12 months without an alternative 
medical cause. 
− A high 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, confirmation with more than one FSH measurement is required. 
− Females on HRT and whose menopausal status is in doubt will be required to use 
one of the non-estrogen 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. 
Male subjects with a female partner who is not of childbearing potential 
Presumptive fertile male subjects who are partnered with women who are not of childbearing 
potential, either post-menopausal or through sterilization or other surgical procedures (e.g., 
hysterectomy) will confirm that their partners have been determined to be sterile or post-
menopausal based on medical judgment; this can be confirmed either by statement or medical 
records, if presented by the subject. 
Men will not be required to use other contraception when sexually active with a sterile or post-
menopausal partner. 
Men who are sexually active with fertile women need to confirm that they will use 2 forms of 
contraception as per protocol. 


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Contraception Guidance: 
Contraceptive use by men or women should be consistent with local regulations regarding the 
methods of contraception for those participating in clinical studies. 
Subjects should use highly effective, double contraception (both male and female partners) 
during the study and 30 days or 5 half-lives (whichever is longer) of study intervention. 
Double contraception is defined as a condom with one of any of the following forms of birth 
control: 
• Birth control pills (The Pill) 
• Depot or injectable birth control 
• Intrauterine Device (IUD) 
• Birth Control Patch (e.g., Othro Evra) 
• NuvaRing® 
• vasectomy for men 
Subjects may not donate sperm or oocytes during the study and for 30 days or 5 half-lives 
(whichever is longer) after last dose of study intervention. 
Rhythm methods are not considered as highly effective methods of birth control. Subject 
abstinence for the duration of the study and 30 days or 5 half-lives (whichever is longer) after 
last dose of study intervention is acceptable if it is the subject’s regular practice. 
Collection of Pregnancy Information 
Male subjects with partners who become pregnant 
• The Investigator will attempt to collect pregnancy information on any male subject’s 
female partner who becomes pregnant while the male subject is in this study. This 
applies only to male subjects who receive study intervention. 
• 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 within 24 hours of learning of the partner’s 
pregnancy. The female partner will also be followed to determine the outcome of the 
pregnancy. Information on the status of the mother and child will be forwarded to the 
Sponsor. Generally, the follow-up will be no longer than 6 to 8 weeks following the 
estimated delivery date. Any termination of the pregnancy will be reported regardless 
of fetal status (presence or absence of anomalies) or indication for the procedure. 
Female subjects who become pregnant 
• Female subjects who become pregnant should discontinue treatment and be 
withdrawn from the study. 
• The Investigator will collect pregnancy information on any female subject who 
becomes pregnant while participating in this study. Information will be recorded on 
the appropriate form and submitted to the Sponsor within 24 hours of learning of a 


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subject's pregnancy. This applies only to female subjects who receive study 
intervention 
• The subject will be followed to determine the outcome of the pregnancy. The 
Investigator will collect follow-up information on the subject and the neonate and the 
information will be forwarded to the Sponsor. Generally, follow-up will not be 
required for longer than 6 to 8 weeks beyond the estimated delivery date. Any 
termination of pregnancy will be reported, regardless of fetal status (presence or 
absence of anomalies) or indication for the procedure. 
• While pregnancy itself is not considered to be an AE or SAE, any pregnancy 
complication or elective termination of a pregnancy will be reported as an AE or 
SAE. A spontaneous abortion is always considered to be an SAE and will be reported 
as such. Any post-study pregnancy related SAE considered reasonably related to the 
study intervention by the Investigator will be reported to the Sponsor as described in 
Section 7.3.4. While the Investigator is not obligated to actively seek this information 
in former study subjects, he or she may learn of an SAE through spontaneous 
reporting. 
 
 


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9.5. 
Appendix 5: Considerations in Response to Coronavirus Disease 
2019 (COVID-19) Pandemic  
As of March 12, 2020, a COVID-19 pandemic has been declared by the World Health 
Organization, leading to implementation of extensive measures by healthcare systems globally to 
limit viral spread, with potential impact on conduct of clinical studies.  
Based on recommendations in the FDA Guidance on Conduct of Clinical Trials of Medical 
Products during COVID-19 Pandemic (FDA, July 2020), the following actions were 
implemented to address potential disruptions to study conduct secondary to COVID-19 infection 
or control measures: 
• Interruption in study intervention administration and other study procedures 
(collectively “study interruption”) will be allowed for up to 4 weeks (+4 days). Any 
interruption lasting ≥ 2 weeks will be considered a study interruption. 
• For subjects who have a study interruption, prior to resuming dosing, the Investigator 
will assess whether the subject is clinically stable to continue participation in the 
study. This assessment will include symptom-directed physical examination, vital 
signs, pregnancy test (local) in WOCBP, and laboratory tests (hematology and 
clinical biochemistry) to be done locally.  
If clinical laboratory tests are not part of the protocol-defined procedures for the first dosing 
visit after study interruption, hematology and clinical biochemistry panels will be obtained 
prior to dosing and sent to the central lab.  
Results from both the local and central labs should be documented in EDC.  
• Upon resumption of dosing, the subject will continue at the next planned visit relative 
to the last visit performed prior to study interruption, and will be administered all 
remaining planned doses.  
• Administration of study intervention by a trained home health care worker will be 
considered on a case-by-case basis if circumstances related to the COVID-19 
pandemic preclude dosing at the study site  
• In cases where a subject is continuing to receive study intervention but COVID-19 
pandemic-related circumstances preclude a visit to the investigative site, remote visits 
(e.g., by telemedicine or phone contact) will be allowed for relevant study procedures.   
• In cases where a subject is continuing to receive study intervention but cannot obtain 
laboratory tests at the investigative site, local laboratory tests may be obtained at a 
certified laboratory. The site should inform the Sponsor about such cases. Local 
analysis can be used for safety decisions. In addition, local labs can be used for trial 
endpoints if samples drawn at the local lab cannot be shipped to the central lab.   
• (For Part 1 only) The Investigator should discuss with the Sponsor on a case-by-case 
basis if circumstances related to the COVID-19 pandemic preclude domiciling around 
the 5th dose (in Cohorts 3 and 4) or the 3rd dose (in Cohorts 5 and 6); with Sponsor 
approval, these visits may be conducted without domiciling. 


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• (For Part 1 only) As MRI facilities may be impacted by circumstances related to the 
COVID-19 pandemic, precluding protocol-defined MRI assessments, all subjects in 
Part 1 who have not yet undergone Day 50 and Day 92/ET MRI assessments will 
undergo Fibroscan assessments on Day 50 and Day 92/ET in addition to the planned 
MRI assessments. 
• End of Treatment may be considered for the following scenarios: 
− For subjects in Part 1 who have reached the Day 57 visit (Visit 17) and subjects in 
Part 2 who have reached the Day 106 visit (Visit 18), an End of Treatment visit 
may be considered instead of the study interruption. The Investigator should 
discuss with the Sponsor about the feasibility of these two options based on the 
potential impact from COVID-19 and/or the control measures in place in their 
location. 
− Study interruptions longer than 4 weeks will lead to discontinuation of treatment 
and the subject’s participation in the study.   
Efforts should be made to obtain End of Treatment assessments in subjects who are not 
willing to return to the study after study interruption, who are deemed not clinically stable to 
continue participation by the Investigator, or who are lost to the study due to disruptions 
because of COVID-19 pandemic. The End of Treatment visit should be scheduled as soon as 
possible in these scenarios, whenever possible.  
• For dose interruptions lasting ≥2 weeks, contact will be established with the study 
subject remotely (e.g., by phone or telemedicine) to obtain information about AEs, 
concomitant medications, or any other update related to the subjects’ safety. 
• Study interruption will be allowed as long as COVID-19 pandemic-related 
circumstances are ongoing, and will not be allowed when these circumstances are no 
longer applicable, as determined by the Sponsor. 
• For newly randomized subjects who have not yet received their first dose at a site that 
can foresee near-term disruption by COVID-19 pandemic, the Investigator should 
discuss with the Sponsor whether the initiation of dosing should be postponed. In this 
situation, study interruption should not apply, and the subject will initiate and finish 
all planned doses when the Investigator deems it is safe to start dosing. 
Changes in study visit schedules, missed visits, or patient discontinuations may lead to missing 
information (e.g., for protocol-specified procedures). Specific information will be captured in the 
case report form that explains the basis of the missing data, including the relationship to 
COVID-19 for missing protocol-specified information (e.g., from missed study visits or study 
discontinuations due to COVID-19). 
 
 


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9.6. 
Appendix 6: Glossary 
Abbreviation Term 
Description 
AAA 
Abdominal aortic aneurysm  
ADA 
Antidrug Antibodies  
ADL 
Activities of Daily Living  
AE 
Adverse events  
AI 
Artificial intelligence 
ALP 
Alkaline phosphatase 
ALT 
Alanine transaminase  
AST 
Aspartate transaminase  
AUC0-tau 
Area under the serum drug concentration by time curve within a dosing 
interval 
BMD 
Bone mineral density  
BMI 
Body mass index  
BUN 
Blood urea nitrogen  
CAD 
Coronary artery disease  
CAP 
Controlled Attenuation Parameter  
CBC 
Complete blood count  
CFR 
Code of Federal Regulations  
CHF 
Congestive heart failure  
CIOMS 
International Organizations of Medical Sciences  
CK 
Cytokeratin  
Cmax  
Maximal observed serum concentrations  
COVID-19 
Coronavirus disease 2019 
CPK 
Creatine phosphokinase 
CRO 
Clinical research organization 
CTCAE 
Common terminology criteria for adverse events 
CTX 
Carboxy-terminal collagen crosslinks 
CV 
Coefficient of variation 
DAA 
Direct-acting antiviral 
DILI 
Drug-induced liver injury 
DNA 
Deoxyribonucleic acid 
DPP-IV 
Dipeptidyl peptidase IV 
ECG 
Electrocardiogram  
eCRF 
Electronic case report form 
eGFR 
Estimated Glomerular filtration rate  
ELF 
Enhanced liver fibrosis  
EOS 
End of study 


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Abbreviation Term 
Description 
ET 
Early Termination  
EU 
European Union  
FDA 
Food and Drug Administration  
FGFR 
Fibroblast growth factor receptor 
FIB4 
Fibrosis-4 
FSH 
Follicle stimulating hormone  
FU 
Follow up  
GCP 
Good Clinical Practice  
GGT 
Gamma-glutamyl transferase  
GI  
Gastrointestinal 
GLP  
Good Laboratory Practice 
GLP-1 
Glucagon-like peptide 1 
HA 
Hyaluronic acid  
HbA1c 
Hemoglobin A1c 
HCC 
Hepatocellular carcinoma  
hCG 
human chorionic gonadotropin  
HCV 
Human coronavirus 
HDL 
High density lipoprotein 
HDL-c 
High density lipoprotein cholesterol 
HIPAA 
Health Insurance Portability and Accountability Act  
HIV 
Human Immunodeficiency Virus  
HOMA-IR 
Method used to quantify insulin resistance 
HRT 
Hormonal replacement therapy  
hsCRP 
High sensitivity C-reactive protein  
IB 
Investigator’s brochure  
ICF 
Informed consent form  
ICH 
Council for Harmonisation  
IEC 
Independent Ethics Committee  
IGF-1 
Insulin-like growth factor-1 
INR 
Normalized ratio  
IRB 
Institutional Review Board  
IRT 
Interactive Response Technology 
IUD 
Intrauterine device  
LDL 
Low density lipoprotein  
LDL-c 
Low density lipoprotein cholesterol 
MAD 
Multiple ascending dose  
MCH 
Mean corpuscular hemoglobin 


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Abbreviation Term 
Description 
MCV 
Mean corpuscular volume 
MedDRA 
Medical Dictionary for Regulatory Activities 
MetS 
Metabolic syndrome  
MRI 
Magnetic resonance imaging  
n 
Number 
NAb 
Neutralizing antibodies 
NAFLD 
Nonalcoholic fatty liver disease  
NAS 
NASH with NAFLD Activity  
NASH 
Nonalcoholic Steatohepatitis  
NCI 
National Cancer Institute 
NOAEL 
No observed adverse effect level  
NYHA 
New York Heart Association 
OGTT 
Oral glucose tolerance test  
OTC 
Over-the-counter  
PBC 
Primary biliary cirrhosis  
P1NP 
N-terminal propeptide of type 1 collagen 
PD 
Pharmacodynamic  
PDFF 
Proton density fat fraction 
PEG 
Polyethylene glycol 
Pro-C3 
N‐terminal propeptide of type III collagen 
PK 
Pharmacokinetic  
PSC 
Primary sclerosing cholangitis  
PT 
Preferred term  
PVD 
Peripheral vascular disease  
QW 
Weekly  
Q2W 
Every 2 weeks 
QRS 
Complex in ECG representing ventricular depolarization 
QTcF 
Fredericia corrected QT interval in ECG 
RBC 
Red blood cell  
RNA 
Ribonucleic acid  
SAD 
Single ascending doses  
SAE 
Serious adverse event 
SAP 
Statistical Analysis Plan  
SAT 
Subcutaneous abdominal fat 
SC 
Subcutaneous  
SD 
Standard deviation 
SMC 
Safety Monitoring Committee  


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Abbreviation Term 
Description 
SoA 
Schedule of Activities  
SOC 
System organ class  
SOP 
Standard Operating Procedure 
SVR 
Sustained viral response 
t½  
Terminal elimination half-life 
T2DM 
Type 2 diabetes mellitus 
TB 
Total bilirubin  
TEAE 
Treatment-emergent adverse event 
TIA 
Transient ischemic attack  
tmax  
Time to achieve Cmax 
TSH 
Thyroxine stimulating hormone 
ULN 
Upper limit of normal  
US 
United States  
VAT 
Visceral abdominal fat 
VCTE 
Vibration-controlled transient elastography 
WBC 
Blood cell count  
WOCBP 
Woman of Childbearing Potential  
 
 


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10. 
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Harrison, S. A., Rinella, M. E., Abdelmalek, M. F., Trotter, J. F., Paredes, A. H., Arnold, H. L., 
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Hashimoto, E., Yatsuji, S., Tobari, M., Taniai, M., Torii, N., Tokushige, K., et al. (2009). 
"Hepatocellular carcinoma in patients with nonalcoholic steatohepatitis." J Gastroenterol 44 
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Isabela Andronescu, C., Roxana Purcarea, M. and Aurel Babes, P. (2018). "The role of 
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Janes, C.H. and Lindor, K.D. (1993). "Outcome of patients hospitalized for complications after 
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Khan, F. Z., Perumpail, R. B., Wong, R. J. and Ahmed, A. (2015). "Advances in hepatocellular 
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Kim, A. M., Somayaji, V. R., Dong, J. Q., Rolph, T. P., Weng, Y., Chabot, J. R., et al. (2017). 
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lipids, bone turnover markers, blood pressure and body weight differently in obese people with 
hypertriglyceridaemia and in non-human primates." Diabetes Obes Metab 19(12): 1762-1772. 
Kleiner, D. E., Brunt, E. M., Van Natta, M., Behling, C., Contos, M. J., Cummings, O. W., et al. 
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disease." Hepatology 41(6): 1313-1321. 
Lee, D. V., Li, D., Yan, Q., Zhu, Y., Goodwin, B., Calle, R., et al. (2014). "Fibroblast growth 
factor 21 improves insulin sensitivity and synergizes with insulin in human adipose stem cell-
derived (hASC) adipocytes." PLoS One 9(11): e111767. 


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Lee, P., Linderman, J., Smith, S., Brychta, R. J., Perron, R., Idelson, C., et al. (2013). "Fibroblast 
growth factor 21 (FGF21) and bone: is there a relationship in humans?" Osteoporos Int 24(12): 
3053-3057. 
Levey, A. S., Stevens, L. A., Schmid, C. H., Zhang, Y. L., Castro, A. F., 3rd, Feldman, H. I., et 
al. (2009). "A new equation to estimate glomerular filtration rate." Ann Intern Med 150(9): 604-
612. 
Li, M., Reynolds, C. M., Segovia, S. A., Gray, C. and Vickers, M. H. (2015). "Developmental 
Programming of Nonalcoholic Fatty Liver Disease: The Effect of Early Life Nutrition on 
Susceptibility and Disease Severity in Later Life." BioMed research international 2015: 437107-
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Lichtinghagen, R., Pietsch, D., Bantel, H., Manns, M. P., Brand, K. and Bahr, M. J. (2013). "The 
Enhanced Liver Fibrosis (ELF) score: normal values, influence factors and proposed cut-off 
values." J Hepatol 59(2): 236-242. 
Loomba, R., Neuschwander-Tetri, B.A., Sanyal, A., Chalasani, N., Diehl, A.M., Terrault, N., et 
al. (2017). "Novel multicenter validation of association between decline in MRI-PDFF and 
histologic response: a secondary analysis of FLINT trial." Poster presented at American 
Association for the Study of Liver Diseases (AASLD). 
Loomba, R., Sanyal, A.J., Kowdley, K.V., Terrault, N., Chalasani, N.P., Abdelmalek, M.F., et al. 
(2019). "Factors associated with histologic response in adult patients with nonalcoholic 
steatohepatits. Gastroenterology 156: 88-95. 
Loomba, R., Sirlin, C.B., Ang, B., Bettencourt, R., Jain, R., Salotti, J., et al. (2015). "Ezetimibe 
for the treatment of nonalcoholic steatohepatitis: assessment by novel magnetic resonance 
imaging and magnetic resonance elastography in a randomized trial (MOZART trial)." 
Hepatology 61(4): 1239-1250. 
Matthews, D. R., Hosker, J. P., Rudenski, A. S., Naylor, B. A., Treacher, D. F. and Turner, R. C. 
(1985). "Homeostasis model assessment: insulin resistance and beta-cell function from fasting 
plasma glucose and insulin concentrations in man." Diabetologia 28(7): 412-419. 
Mittal, S., El-Serag, H. B., Sada, Y. H., Kanwal, F., Duan, Z., Temple, S., et al. (2016). 
"Hepatocellular Carcinoma in the Absence of Cirrhosis in United States Veterans is Associated 
With Nonalcoholic Fatty Liver Disease." Clin Gastroenterol Hepatol 14(1): 124-131.e121. 
Mu, J., Pinkstaff, J., Li, Z., Skidmore, L., Li, N., Myler, H., et al. (2012). "FGF21 analogs of 
sustained action enabled by orthogonal biosynthesis demonstrate enhanced antidiabetic 
pharmacology in rodents." Diabetes 61(2): 505-512. 
Nielsen, M. J., Veidal, S. S., Karsdal, M. A., Orsnes-Leeming, D. J., Vainer, B., Gardner, S. D., 
et al. (2015). "Plasma Pro-C3 (N-terminal type III collagen propeptide) predicts fibrosis 
progression in patients with chronic hepatitis C." Liver Int 35(2): 429-437. 
Ogawa, Y., Kurosu, H., Yamamoto, M., Nandi, A., Rosenblatt, K. P., Goetz, R., et al. (2007). 
"BetaKlotho is required for metabolic activity of fibroblast growth factor 21." Proc Natl Acad 
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Owen, B. M., Bookout, A. L., Ding, X., Lin, V. Y., Atkin, S. D., Gautron, L., et al. (2013). 
"FGF21 contributes to neuroendocrine control of female reproduction." Nat Med 19(9): 1153-
1156. 
Park, J. G., Xu, X., Cho, S., Hur, K. Y., Lee, M. S., Kersten, S., et al. (2016). "CREBH-FGF21 
axis improves hepatic steatosis by suppressing adipose tissue lipolysis." Sci Rep 6: 27938. 
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percutaneous liver biopsy. A multicentre retrospective study on 68,276 biopsies." J Hepatol 2: 
165-173. 
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psychiatry 168(12): 1266-1277. 
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2263-2273. 
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Association for the Study of Liver Diseases. (2009). "Liver biopsy." Hepatology 49: 1017-1044. 
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placebo-controlled, phase 2a trial." Lancet 392(10165): 2705-2717. 
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"Complication rate of percutaneous liver biopsies among persons with advanced chronic liver 
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nonalcoholic fatty liver disease." Clin Gastroenterol Hepatol 10(8): 837-858. 
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5832. 
 

SAP_001.pdf: 
BIO89-100-002
Page 1 of 52
SAP Version 1.0 (Final) – 21AUG2020
Statistical Analysis Plan
Protocol BIO89-100-002
A Randomized, Double-Blind, Placebo-Controlled, Multiple Ascending Dose Study to Evaluate 
the Safety, Tolerability and Pharmacokinetic Properties of BIO89-100 Administered 
Subcutaneously in Subjects with Nonalcoholic Steatohepatitis (NASH) or with Nonalcoholic 
Fatty Liver Disease (NAFLD) and at High Risk of NASH
Phase 1b
SAP Version:
1.0 (Final)
Date:
21AUGUST2020
Prepared by:
On Behalf of ProSciento Inc.
NCT #: NCT04048135


BIO89-100-002
Page 2 of 52
SAP Version 1.0 (Final) – 21AUG2020
Statistical Analysis Plan
Protocol BIO89-100-002
A Randomized, Double-Blind, Placebo-Controlled, Multiple Ascending Dose Study to Evaluate 
the Safety, Tolerability and Pharmacokinetic Properties of BIO89-100 Administered 
Subcutaneously in Subjects with Nonalcoholic Steatohepatitis (NASH) or with Nonalcoholic 
Fatty Liver Disease (NAFLD) and at High Risk of NASH
Phase 1b
SAP Version:
1.0 (Final)
Date:
21AUG2020
This statistical analysis plan has been reviewed and approved by:
Date:
89bio LTD
Date:
89bio LTD
Date:
89bio LTD
Date:
Pharmapace, On Behalf of ProSciento Inc.
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Table of Contents
1.
Introduction ............................................................................................................................. 9 
2.
Study Objectives...................................................................................................................... 9 
2.1.
Primary Objectives............................................................................................................... 9 
2.2.
Secondary Objectives........................................................................................................... 9 
2.3.
Exploratory Objectives ........................................................................................................ 9 
3.
Study Overview....................................................................................................................... 9 
3.1.
Study Design........................................................................................................................ 9 
3.2.
Study Activities.................................................................................................................. 11 
3.3.
Randomization Schedule and Blinding Procedures........................................................... 11 
3.4.
Stratification Factors.......................................................................................................... 11 
3.5.
Sample Size and Power...................................................................................................... 11 
3.6.
Interim Analysis................................................................................................................. 12 
3.7.
Safety Monitoring Committee ........................................................................................... 12 
4.
Study Endpoints..................................................................................................................... 12 
4.1.
Primary Endpoints ............................................................................................................. 12 
4.1.1.
Safety Endpoints ............................................................................................................ 12 
4.1.2.
Pharmacokinetic Endpoints............................................................................................ 12 
4.2.
Secondary Endpoints ......................................................................................................... 12
4.3.
Other Safety Endpoints...................................................................................................... 14 
4.4.
Exploratory Endpoints ....................................................................................................... 14 
5.
Definitions ............................................................................................................................. 15 
5.1.
Definition of Study Day..................................................................................................... 15 
5.2.
Handling of Missing or Partial Dates................................................................................. 15 
5.2.1.
Adverse Event Date Imputations.................................................................................... 15 
5.2.2.
Medication/Procedure Date Imputations........................................................................ 16 
5.3.
Definition of Study Baseline.............................................................................................. 17 
5.3.1.
Study Baseline of ALT and AST ................................................................................... 17 
5.3.2.
Study Baseline for Lipids............................................................................................... 18 
5.3.3.
Study Baseline for Vital Signs ....................................................................................... 18 
5.4.
Definition of Duration of Exposure................................................................................... 18 


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5.5.
Definition of Age............................................................................................................... 19 
5.6.
Dose Compliance............................................................................................................... 19 
5.7.
Definition of Pre-treatment Medication and Pre-treatment Procedures ............................ 19 
5.8.
Definition of Baseline Medication..................................................................................... 19 
5.9.
Definition of Concomitant Medication and Concomitant Procedures............................... 19 
5.10.
Definition of Randomized Treatment Group ................................................................. 19 
5.11.
Definition of Actual Treatment Group........................................................................... 19 
5.12.
Definition of Treatment Emergent Adverse Event ........................................................ 20 
5.17.
Adipo-IR......................................................................................................................... 21 
5.18.
Estimated Glomerular Filtration Rate (CKD-EPI)......................................................... 21 
5.19.
Definition of Study Interruption..................................................................................... 21 
5.20.
Definition of Actual Time.............................................................................................. 21 
6.
Potential Covariates............................................................................................................... 21 
8.
Analysis Populations ............................................................................................................. 22 
8.1.
Screened Analysis Set........................................................................................................ 22 
8.2.
Randomized Analysis Set .................................................................................................. 22 
8.3.
Safety Analysis Set ............................................................................................................ 22 
8.4.
Pharmacokinetics Analysis Set.......................................................................................... 23 
8.5.
Pharmacodynamics Analysis Set....................................................................................... 23 
8.6.
Pharmacodynamics Analysis Set - MRI ............................................................................ 23 
8.7.
Pharmacodynamics Analysis Set - Fibroscan.................................................................... 23 
9.
Statistical Methods of Data Analyses.................................................................................... 23 
9.1.
General Considerations...................................................................................................... 23 
9.1.1.
Statistical Notation and Presentation.............................................................................. 23 
9.1.2.
Statistical Modeling Methods......................................................................................... 24 
9.1.3.
Analysis Visits and Visit Windowing............................................................................ 24 


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9.1.4.
Assessments Below and Above the Limit of Quantification ......................................... 26 
9.2.
Pharmacokinetic Analyses................................................................................................. 26 
9.2.1.
Analysis of Serum Concentrations of BIO89-100 ......................................................... 26 
9.2.2.
Analysis of PK Parameters............................................................................................. 26 
9.3.
Study Subjects.................................................................................................................... 27 
9.3.1.
Subject Disposition ........................................................................................................ 27 
9.3.2.
Demographics and Baseline Characteristics .................................................................. 27 
9.3.3.
Biopsy History................................................................................................................ 28 
9.3.4.
Medical History.............................................................................................................. 28 
9.3.5.
Inclusion/Exclusion Criteria........................................................................................... 28 
9.3.6.
Investigational Product Administration ......................................................................... 28 
9.3.7.
Concomitant Medication................................................................................................ 28 
9.3.8.
Concomitant Procedures ................................................................................................ 29 
9.3.9.
Protocol Deviations........................................................................................................ 29 
9.4.
Pharmacodynamic Analyses.............................................................................................. 29 
9.4.1.
Analyses of Anthropomorphic Measurements............................................................... 29 
9.4.2.
Analyses of Laboratory Parameters ............................................................................... 29 
9.4.2.1.
Lipids.......................................................................................................................... 29
9.4.2.2.
Metabolic Biomarkers ................................................................................................ 30 
9.4.2.3.
Liver Biomarkers........................................................................................................ 30 
9.4.2.3.1.
Sensitivity Analysis for Liver Biomarkers ............................................................. 30 
9.4.2.4.
Suspected Drug Induced Liver Injury ........................................................................ 31 
9.4.3.
Analyses of MRI Measures............................................................................................ 32 
9.4.3.1.
Responder Analysis .................................................................................................... 32 
9.4.3.2.
Supportive Analysis.................................................................................................... 33 
9.4.3.2.1.
Multiple Imputation Using Control Based Pattern-Mixture Model........................ 33 
9.4.3.3.
Correlation Analysis................................................................................................... 33 
9.4.3.4.
Non-Parametric Analysis............................................................................................ 33 


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9.5.
Safety Analyses.................................................................................................................. 35 
9.5.1.
Adverse Events............................................................................................................... 35 
9.5.2.
Clinical Laboratory Evaluations..................................................................................... 36 
9.5.3.
Physical Examination Findings...................................................................................... 36 
9.5.4.
Vital Signs...................................................................................................................... 36 
9.5.5.
12-Lead Electrocardiograms .......................................................................................... 37 
9.5.6.
Columbia-Suicide Severity Rating Scale (C-SSRS)...................................................... 37 
9.5.7.
Immunogenicity Analyses.............................................................................................. 38 
10.
Changes to Planned Analysis in the Protocol .................................................................... 38 
11.
Statistical Software ............................................................................................................ 38 
12.
References.......................................................................................................................... 38 
13.
Appendix............................................................................................................................ 39 
13.1.
Sample SAS Code.......................................................................................................... 51 
Table 1: Dose Escalation Cohorts................................................................................................. 10 
Table 2 Schedule of Activities for Cohorts 1-4 ............................................................................ 39 
Table 3 PK Sample Collection for Cohorts 1-4 – QW Dosing Interval ....................................... 44 
Table 4 Schedule of Activities for Cohorts 5 and 6...................................................................... 45 
Table 5 PK Sample Collection for Cohorts 5 and 6 – Q2W Dosing Interval............................... 50 


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SAP Version 1.0 (Final) – 21AUG2020
Abbreviations
ADA
anti-drug antibodies 
AEs
adverse events
ALT
alanine transaminase
AST
aspartate transaminase  
ATC
anatomic therapeutic chemistry
AUC0-tau
area under the curve since last dose 
BMI
body mass index
CBC
complete blood count 
CK-18
cytokeratin-18
CAP
controlled attenuation parameter
Cmax
maximum concentration 
COVID-19
Coronavirus disease 2019
C-SSRS
columbia-suicide severity rating scale 
CTCAE
common terminology criteria for adverse events  
CTX
carboxy-terminal collagen crosslinks  
CV%
coefficient of variation 
DNA
deoxyribonucleic acid
ECG
electrocardiogram  
eCRF
electronic Case Report Form
eDISH
evaluation of drug induced serious hepatotoxicity
eGFR
estimated glomerular filtration rate
ELF
enhanced liver fibrosis  
EOS
end of study
ET
early termination
FFA
free fatty acids  
FGF21
fibroblast growth factor 21 
FU
follow-up
HA
Hyaluronic acid
HbA1c
hemoglobin a1c  
HDL-c
high density lipoprotein  
HIV
human immunodeficiency virus  
HOMA-IR
homeostatic model of assessment of insulin resistance 
hsCRP
high-sensitivity c-reactive protein 
IGF-1
insulin-like growth factor-1
IP
investigational product
LDL-c
low density lipoprotein 
LS
least squares 
MAD
multiple ascending dose  
MAR
missing at random
MedDra
medical dictionary for regulatory activities 
MMRM
mixed model repeated measures  
MNAR
missing not at random


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MRI-PDFF
magnetic resonance imaging – proton density fat fraction   
n
sample size
NAFLD
nonalcoholic fatty liver disease 
NASH
nonalcoholic steatohepatitis  
non-HDL
non-high density lipoprotein  
OGTT
oral glucose tolerance test  
P1NP
n-terminal propeptide of type 1 collagen  
PD
pharmacodynamics  
PEG
polyethylene glycol 
PIIINP
Amino-terminal propeptide of type III procollagen
PMM
pattern-mixture model
Pro-C3
n-terminal propeptide of type iii collagen  
PT
preferred term 
Q2W
every two weeks 
QTcF
corrected qt interval by Fridericia
QW
weekly  
SAEs
serious adverse events 
SAP
statistical analysis plan  
SC
subcutaneous  
SD
standard deviation
SE
standard error  
SMC
safety monitoring committee 
SOC
system organ class  
t1/2
half-life
TB
total bilirubin 
TEAE
treatment-emergent adverse event 
TIMP-1
tissue inhibitor of metalloproteinase 1
tmax
time to maximum concentration 
TSH
thyroid stimulating hormone
ULN
upper limit of normal  
VCTE
vibration-controlled transient elastography
WHO
world health organization  
WOCBP
women of child-bearing potential


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1. Introduction
This statistical analysis plan (SAP) describes the statistical methods and procedures to be 
implemented in clinical study BIO89-100-002. This statistical analysis plan is based on study 
protocol BIO89-100-002 Version 3.0 dated 25 March 2020. If the data suggest and warrant it, 
deviations from this plan will be considered. However, any deviations from the SAP must be 
substantiated by sound statistical rationale and documented in the final clinical study report.
2. Study Objectives
2.1. Primary Objectives
The primary objectives of the study are:
x
To evaluate the safety and tolerability of ascending multiple subcutaneous (SC) injections 
of BIO89-100 in subjects with nonalcoholic steatohepatitis (NASH) or who have 
nonalcoholic fatty liver disease (NAFLD) and at a high risk of NASH.
x
To characterize BIO89-100 pharmacokinetics
2.2. Secondary Objectives
The secondary objectives of the study are:
x
To evaluate the immunogenicity of BIO89-100 as measured by presence of anti-drug 
antibodies (ADA)
x
To characterize biomarkers, pharmacodynamics (PD) profile and biological activity of 
BIO89-100 administered at ascending doses and with both weekly (QW) and every two 
weeks (Q2W) dosing intervals
x
To evaluate the time, dose, and exposure relationship of BIO89-100 on biological 
activity, as assessed by biomarkers and PD
2.3. Exploratory Objectives
The exploratory objective of the study is:
 
3. Study Overview
3.1. Study Design
Study BIO89-100-002 is a randomized, double-blind, placebo-controlled, multiple ascending 
dose (MAD) study to evaluate the safety, tolerability, PK and PD profiles and immunogenicity of 
BIO89-100 administered SC in approximately 83 subjects with NASH, or with NAFLD who are 
at a high risk of NASH. This multi-site study will consist of 6 cohorts, and will evaluate 2 dosing 
schedules, weekly (QW; cohorts 1 to 4) and every 2 weeks (Q2W; Cohorts 5 and 6).


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Table 1: Dose Escalation Cohorts
Cohort
Randomized 
Group/Dose Level
Frequency and Route of 
Administration
Number of Subjects
BIO89-100
Placebo
1
BIO89-100 3 mg QW
Weekly (QW), SC to abdomen 
(1 injection) Days 1, 8, 15, 22, 
29, 36, 43, 50, 57, 64, 71, 78, 
and 85
Total doses (initial 4 weeks + 8 
week extension): 5 + 8
6
2
2
BIO89-100 9 mg QW
QW, SC to abdomen (1 
injection) Days 1, 8, 15, 22, 29, 
36, 43, 50, 57, 64, 71, 78, and 85
Total doses (initial 4 weeks + 8 
week extension): 5 + 8
12
3
3
BIO89-100 18 mg QW
QW, SC to abdomen (1 
injection) Days 1, 8, 15, 22, 29, 
36, 43, 50, 57, 64, 71, 78, and 85
Total doses (initial 4 weeks + 8 
week extension): 5 + 8
14
4
4
BIO89-100 27 mg QW
QW, SC to abdomen (1 
injection) Days 1, 8, 15, 22, 29, 
36, 43, 50, 57, 64, 71, 78, and 85
Total doses (initial 4 weeks + 8 
week extension): 5 + 8
9
3
5
BIO89-100 18 mg Q2W
Every 2 weeks (Q2W), SC to 
abdomen (1 injection)
Days 1, 15, 29, 43, 57, 71 and 
85.
Total doses (initial 4 weeks + 8 
week extension): 3 + 4
14
4
6
BIO89-100 36 mg Q2W
Q2W, SC to abdomen (2 
injections)
Days 1, 15, 29, 43, 57, 71 and 
85.
Total doses (initial 4 weeks + 8 
week extension): 3 + 4
9
3
The study will include a Screening period, a Treatment period, and a Follow-up period. The 
treatment duration will be 12 weeks, consisting of a 4-week study for initial safety assessment 
(primary endpoint) with an 8-week extension phase for efficacy and safety assessments. The 
study includes an individual safety checkpoint after 4 weeks of treatment (through Day 30), to 


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ensure that only individuals with an acceptable safety profile will proceed to the 8-week
extension phase.
There will be 2 dose escalation decisions. After Cohort 1 completes the Day 36 visit, the Safety 
Monitoring Committee (SMC) will decide whether subjects can be randomized into Cohorts 2 
and 5 (both to start concurrently). After at least eight subjects from both Cohort 2 and Cohort 5, 
including at least 1 subject on placebo in each cohort, complete the Day 36 visit, the SMC will 
decide whether subjects can be randomized into Cohorts, 3,4 and 6 (all to start concurrently).
3.2. Study Activities
Please refer to the appendix for the schedule of activities and pharmacokinetic sample collection
schedules:
x
Table 1 Schedule of Activities for Cohorts 1-4
x
Table 2 PK Sample Collection for Cohorts 1-4 – QW Dosing Interval
x
Table 3 Schedule of Activities for Cohorts 5 and 6
x
Table 4 PK Sample Collection for Cohorts 5 and 6 – Q2W Dosing Interval
3.3. Randomization Schedule and Blinding Procedures
Eligible subjects will be randomized in the order they are enrolled into the study. For Cohort 1,
eight eligible subjects will be randomized in a 3:1 ratio to active (BIO89-100): control (placebo). 
For Cohort 2, fifteen subjects will be randomized in a 4:1 ratio to active: control. For cohorts 4 
and 6, twelve eligible subjects will be randomized in each cohort in a 3:1 ratio to active: control. 
For Cohorts 3 and 5, eighteen subjects will be randomized in each cohort in a ratio of 7:2 active:
control, stratified by biopsy confirmed NASH with fibrosis status F1, F2 or F3 (Yes, No), with 
each stratum of nine subjects in each cohort.
3.4. Stratification Factors
Cohorts 3 and 5, will be stratified by biopsy confirmed NASH with fibrosis status F1, F2 or F3 
(Yes, No).
3.5. Sample Size and Power
No formal sample size calculation was performed for the primary endpoint(s) as the total of 
approximately 83 subjects, consisting of 6:2 (Cohort 1), 12:3 (Cohort 2), 9:3 (Cohorts 4 and 6) or 
14:4 (Cohorts 3 and 5) receiving active: placebo, respectively is considered adequate to inform 
on the endpoints for this Phase 1b study.
A power assessment based on magnetic resonance imaging – proton density fat fraction (MRI-
PDFF) endpoint (percent change from baseline to End of Treatment in liver fat content, an 
important biomarker) shows that 9, 12 and 14 subjects in a dose cohort compared to the pooled 
19 placebo subjects will provide 
 power, respectively, to detect a 
difference in mean percentage change from baseline between treatment groups of 
, assuming 


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a standard deviation of 
 in each group. Calculation was based on two-sample one-sided t-test 
with alpha at 0.05 (e.g., two-sided alpha at 0.10).
Subjects who withdraw from the study before the Day 50 assessments may be replaced at the 
discretion of the Sponsor.
3.6. Interim Analysis
No efficacy interim analysis is planned. Sponsor may perform administrative interim analyses to 
support objectives such as study planning and regulatory interactions.
3.7. Safety Monitoring Committee
A Safety Monitoring Committee (SMC), comprised of a Principal Investigator participating in 
the study, the Clinical Research Organization (CRO) Medical Monitor and the Sponsor Medical 
Monitor, will review blinded safety data for dose escalation decisions. The operations of the 
SMC will be defined in the SMC Charter.
4. Study Endpoints
4.1. Primary Endpoints
This study has primary safety and primary pharmacokinetic endpoints.
4.1.1. Safety Endpoints
The primary safety endpoints are:
x
Subject incidence of treatment emergent adverse events (TEAEs)
x
Subject incidence of TEAEs by severity
x
Subject incidence of serious TEAEs
x
Number of subjects who discontinued due to TEAEs
x
Number of subjects who discontinued due to related TEAEs
4.1.2. Pharmacokinetic Endpoints
The primary pharmacokinetic endpoints are:
x
Maximum concentration (Cmax) on Day 1
x
Cmax on Day 29
x
Area under the curve since last dose (AUC0-tau) on Day 1
x
AUC0-tau on Day 29
x
Time to maximum concentration (tmax) on Day 1
x
tmax on Day 29
x
Half-life (t1/2) on Day 1
x
t1/2 on Day 29
4.2. Secondary Endpoints
The secondary immunogenicity endpoints are:


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x
Incidence and characteristics of ADA after dosing (e.g., titer and/or binding specificity, to 
the fibroblast growth factor 21 (FGF21) and polyethylene glycol (PEG) part of BIO89-
100 and neutralizing immunogenicity).
x
Impact/correlation of the presence of ADAs on serum BIO89-100 concentrations and 
clinical safety.
The secondary pharmacodynamics endpoints are:
Anthropomorphic measurements:
x
Change from baseline and percent change from baseline in body weight over time.
x
Change from baseline and percent change from baseline in body mass index (BMI) over 
time.
Laboratory parameters:
x
Change from baseline and percent change from baseline in triglycerides over time
x
Change from baseline and percent change from baseline in non-high density lipoprotein 
(non-HDL) cholesterol over time
x
Change from baseline and percent change from baseline in high density lipoprotein 
(HDL-c) over time
x
Change from baseline and percent change from baseline in low density lipoprotein (LDL-
c) over time
x
Change from baseline and percent change from baseline in hemoglobin A1c (HbA1c)
over time
x
Change from baseline and percent change from baseline in homeostatic model of 
assessment of insulin resistance (HOMA-IR) over time
Liver function tests:
x
Change from baseline and percent change from baseline in alanine transaminase (ALT) 
over time
x
Change from baseline and percent change from baseline in aspartate transaminase (AST)
over time
x
Change from baseline and percent change from baseline in adiponectin over time
x
Change from baseline and percent change from baseline in N-terminal propeptide of type 
III collagen (Pro-C3) over time
x
Change from baseline and percent change from baseline in free fatty acids (FFA) over 
time
x
Change from baseline and percent change from baseline in adipo-IR (fasting free fatty 
acids × fasting insulin) over time
Imaging measures:
x
Change from baseline and percent change from baseline in MRI-PDFF over time


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SAP Version 1.0 (Final) – 21AUG2020
x
Percentage reduction from baseline in MRI-3'))DW'D\50 and Day 92
x
MRI-PDFF < 5% at Day 50 and Day 92
4.3. Other Safety Endpoints
The other safety endpoints are:
x
Change from baseline in vital signs
x
Incidence and shifts of clinically significant vital signs
x
Incidence of clinically significant physical examination findings
x
Incidence and shifts of clinically significant electrocardiograms (ECG)
x
Incidence and shifts of clinically significant laboratory abnormalities
x
Change from baseline in safety laboratory evaluations including complete blood count 
(CBC), blood biochemistry, cortisol and urinalysis over time
4.4. Exploratory Endpoints
 
 
 
 
 
 
 


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SAP Version 1.0 (Final) – 21AUG2020
 
 
5. Definitions
5.1. Definition of Study Day
The day subjects receive their first dose of investigational product will be considered Study Day 
1. Study day will be defined by the number of days from Study Day 1. If index date is prior to 
Study Day 1, study day will be defined as the index date minus the Study Day 1 date. If index 
date is on or after Study Day 1 date, the study day will be defined as index date minus the Study 
Day 1 date plus 1 day. If a subject is randomized but never dosed, study day will be calculated 
using the randomization date and randomization date will be set to Study Day 1.
For subjects with study interruptions due to COVID-19, post-interruption study day will be 
calculated as: (Index date) – (the date of Study Day 1) + 1 – (the interruption duration as defined 
in the study interruption CRF). Visits post-interruption due to COVID-19 will use this corrected 
study day for visit windowing.
5.2. Handling of Missing or Partial Dates
Date imputation will only be used for determining treatment-emergent status for adverse events 
or identifying concomitant medications. Actual data values as they appear in the database will be 
shown in the subject level listings.
5.2.1. Adverse Event Date Imputations
In cases of incomplete dates for adverse events, the missing components will be assumed as the 
most conservative values as possible. The imputation rules described below will attempt to 
conservatively capture AEs with missing onset dates as treatment-emergent, unless information 


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SAP Version 1.0 (Final) – 21AUG2020
of AE stop date/time was prior to first dose date, which would automatically disqualify the 
treatment-emergent status. The original values with no imputation applied will be presented in 
the data listings.
x
If “day” is the only missing field, impute the “day” as the day of Study Day 1 date if the 
month and year of Study Day 1 are equal to the AE start month and year; otherwise, 
impute the “day” as the first day of the AE start month.
x
If “day” and “month” are missing, impute the “day” and “month” as the day and month 
of the first dose date if “year” is the same as the year of Study Day 1; otherwise, impute 
January 1 of the non-missing year.
x
Missing time will not be imputed. If “time” is missing and the start date or imputed start 
date is the same as the first dose date, then the time is assumed to be after the dose time 
so that the event will be classified as treatment emergent.
x
If the start date is completely missing:
o
and from the end date (either complete or partial date) it can be deduced to be 
prior to the first dose date, then the AE will not be assigned as a treatment-
emergent adverse event (TEAE).
o
and from the end date (either complete or partial date) it cannot be deduced to be 
prior to the first dose date, then the AE will be assigned as a TEAE.
o
Start dates that are completely missing will not be imputed.
5.2.2. Medication/Procedure Date Imputations
Imputation of partial start and end dates for medications/procedures is done for the purpose of 
classifying concomitant, pre-treatment, and baseline medications/procedures. The original values 
with no imputation applied will be presented in the data listings.
For medications/procedures with completely missing start dates, no imputation will be 
performed.
For medications/procedures with partial start dates:
x
If “day” is the only missing field, impute the “day” as the first of the month.
x
If “day” and “month” are the missing fields, impute the “day” and “month” to January 1.
x
If only day is missing, and month and year are the same as first dose date then consider as 
concomitant. For a medication/procedure that is not checked as ongoing and the start date 
is completely missing:
o
and from the end date (either complete, partial or completely missing) it can be 
deduced to be prior to the first dose date of randomized investigational product, 
then consider as pre-treatment.
o
Otherwise, consider as concomitant.
For medications/procedures with completely missing end dates and ongoing is not checked:
x
End date will not be imputed, furthermore:


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SAP Version 1.0 (Final) – 21AUG2020
o
If start date is on or after Study Day 1, then consider as concomitant 
medication/procedure
o
If start date is prior to Study Day 1, or start date is also completely missing, then 
consider as pre-treatment medication/procedure
o
If start date is on Study Day 1, then consider as baseline medication/procedure
For medications/procedures with partial end dates:
x
If “day” is the only missing field, and if month and year are the same as the start date and 
ongoing is not checked, then impute “day” as the start date “day”; otherwise impute as 
the last day of the month.
x
If “day” and “month” are the missing fields, if year is the same as the start date and 
ongoing is not checked, then impute “day” and month as the start date day and start date 
month; otherwise impute as December 31.
For medications/procedures that have both completely missing start dates and end dates, no 
imputation will be made; these medications/procedures will be assumed to be concomitant.
5.3. Definition of Study Baseline
For study measurements (safety labs, PD, biomarkers and exploratory labs) except ALT or AST,
lipids, and vital signs, each subject’s baseline value for that measurement will be defined as the
latest pre-dose value. This will be determined by chronologically sorting all scheduled and 
unscheduled values prior to first dose date and time and taking the latest value unless noted 
otherwise. Subjects who are randomized but are not dosed will use their latest measurement as 
baseline up to Study Day 1.
Baseline will be presented with each post-baseline change in the summary tables to allow easier 
review.
5.3.1. Study Baseline of ALT and AST
For ALT and AST, the baseline value will be defined as the average of all values performed 
during screening and Study Day 1 based on the following table. The baseline for ALT and AST 
are calculated and captured in the eCRF.


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ALT or AST Screening Assessments
Day 1 ALT
or AST 
assessment
Baseline value
Assessment 1
Assessment 2
Assessment 3 
(if applicable)
Normal
Normal
Not applicable
Any
Average of Assessment 1, 
Assessment 2 and Day 1 
(3 tests) 
Normal
Abnormal and 
LQFUHDVH
from Assessment 
1
Not applicable
Any
Average of Assessment 1, 
Assessment 2 and Day 1 
(3 tests) 
Normal
Abnormal and 
>40% increase 
from Assessment 
1
Normal or 
% increase 
from 
Assessment 1
Any
Average of Assessment 1, 
Assessment 2, Assessment 
3 and Day 1 (4 tests) 
Normal
Abnormal and 
>40% increase 
from Assessment 
1
Abnormal and 
>40% increase 
from 
Assessment 1
Not 
applicable, 
subject 
excluded.
Not applicable, subject 
excluded.
Abnormal
LQFUHDVH
from Assessment 
1
Not applicable
Any
Average of Assessment 1, 
Assessment 2 and Day 1 
(3 tests) 
Abnormal
>40% increase 
from Assessment 
1
LQFUHDVH
from 
Assessment 1
Any
Average of Assessment 1, 
Assessment 2, Assessment 
3 and Day 1 (4 tests) 
Abnormal
>40% increase 
from Assessment 
1
>40% increase 
from 
Assessment 1
Not 
applicable, 
subject 
excluded.
Not applicable, subject 
excluded.
5.3.2. Study Baseline for Lipids
For lipids, the baseline will be determined as the average of all pre-dose measurements within 30 
days prior to Study Day 1, e.g. measurements taken at -6WXG\'D\3UH-dose).
5.3.3. Study Baseline for Vital Signs
For vital signs of blood pressure and pulse, measurements will be taken in duplicate starting from
randomization. Baseline for these measures should be the last average (of the duplicate 
measurements) on or prior to Study Day 1 and prior to first dose if subject is dosed.
5.4. Definition of Duration of Exposure
The intended duration of exposure for QW dosing will be defined as the date of the last dose of 
investigational product minus Study Day 1 date plus 8 days. The intended duration of exposure 
for Q2W dosing will be defined as the date of the last dose of investigational product minus 
Study Day 1 date plus 15 days.


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SAP Version 1.0 (Final) – 21AUG2020
5.5. Definition of Age
Age will be defined as the duration in days of date of signed informed consent or index date
minus the birthdate divided by 365.25. Index date refers to specific visits if age is needed at 
specific visits (for example age at specific visits will be used when deriving FIB-4 score or 
eGFR).
5.6. Dose Compliance
Dose compliance percentage will be the actual number of received doses divided by the expected 
number of doses to be received up to EOT. Partial doses will contribute to the actual number of 
received doses.
5.7. Definition of Pre-treatment Medication and Pre-treatment Procedures
Pre-treatment medications are defined as any medication that start and stop being taken prior to 
Study Day 1.
Pre-treatment procedures are defined as any procedure that were started and completed prior to 
Study Day 1.
5.8. Definition of Baseline Medication
Medications that have Study Day 1 within the duration of exposure will be considered baseline 
medications.
5.9. Definition of Concomitant Medication and Concomitant Procedures
Concomitant medications are defined as any medication that is taken any day on or after Study 
Day 1. This includes medications that were started prior first dose and that were continued to be 
taken after Study Day 1.
Concomitant procedures are defined as any procedures that are performed any day on or after 
Study Day 1. This includes procedures that were started prior first dose and that continued into 
Study Day 1.
5.10.
Definition of Randomized Treatment Group
The definition of randomized treatment group, also known as the planned treatment group, will 
be the treatment group that a subject was randomized to. Placebo from each cohort will be 
pooled. The randomized treatment groups for summary purpose are: BIO89-100 3 mg QW, 
BIO89-100 9 mg QW, BIO89-100 18 mg QW, BIO89-100 27 mg QW, BIO89-100 18 mg Q2W, 
BIO89-100 36 mg Q2W, and Placebo.
5.11.
Definition of Actual Treatment Group
The definition of actual treatment group will be the assigned treatment group based on what the 
subject actually received. If a subject receives at least one dose of BIO89-100, then that subject 
will be considered a subject of the active treatment group of the cohort that they were 
randomized to. If a subject receives only placebo doses, then their actual treatment group will be 


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SAP Version 1.0 (Final) – 21AUG2020
placebo group. Placebo from each cohort will be pooled. The actual treatment groups for 
summary purpose are: BIO89-100 3 mg QW, BIO89-100 9 mg QW, BIO89-100 18 mg QW, 
BIO89-100 27 mg QW, BIO89-100 18 mg Q2W, BIO89-100 36 mg Q2W and Placebo.
5.12.
Definition of Treatment Emergent Adverse Event
Treatment-emergent adverse events will be defined as those occurring at or after the time of the 
first administration of investigational product, eg, first dose date and time, through study 
termination, or existing prior to the time of and worsening after the time of the first 
administration of investigational product. Adverse events with onset prior to the first 
administration of investigational product that end before Study Day 1 or do not worsen after 
Study Day 1 will be classified as pre-treatment.


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SAP Version 1.0 (Final) – 21AUG2020
5.17.
Adipo-IR
Adipo-IR will be calculated by multiplying fasting insulin (uU/mL) with the fasting free fatty 
acids (mmol/L) concentrations.
5.18.
Estimated Glomerular Filtration Rate (CKD-EPI)
The eGFR using the CKD-EPI equation will be derived for post-baseline visits. Baseline eGFR 
will not be derived and will come from the source data.
For Males:
eGFR[mL/min/1.73mଶ]
= 141
כ min ൬Serum Creatinine[mg/dL]
0.9
, 1൰
ି.ସଵଵ
כ max ൬ܵ݁ݎݑ݉ ܥݎ݁ܽݐ݅݊݅݊݁[mg/dL]
0.9
, 1൰
ିଵ.ଶ଴ଽ
 כ 0.993஺௚௘[௬௘௔௥௦]
כ 1.159[݂݅ ܾ݈ܽܿ݇]
For Females:
eGFR[mL/min/1.73mଶ]
= 141
כ min ൬Serum Creatinine[mg/dL]
0.7
, 1൰
ି.ଷଶଽ
כ max ൬ܵ݁ݎݑ݉ ܥݎ݁ܽݐ݅݊݅݊݁[mg/dL]
0.7
, 1൰
ିଵ.ଶ଴ଽ
 כ 0.993஺௚௘[௬௘௔௥௦] כ 1.018
כ 1.159[݂݅ ܾ݈ܽܿ݇]
5.19.
Definition of Study Interruption
Study interruption will be defined as aQ\LQWHUUXSWLRQODVWLQJZHHNV and allowed up to 4 
weeks (+4 days). that is due to any causes related COVID-19 pandemic and will require extra 
handling when deriving analysis visits.
5.20.
Definition of Actual Time
For PK analysis, actual time will be defined as:
Actual Time=Actual Sample Collection Time - Actual Last Dosing Time (before the 
corresponding sample collection)
6. Potential Covariates
The stratification factor of biopsy confirmed NASH with fibrosis status (F1, F2, F3) Yes or No 
will be considered as a potential covariate to be adjusted for in the statistical models.


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SAP Version 1.0 (Final) – 21AUG2020
 
Subgroup analyses will be descriptive in nature.
8. Analysis Populations
The number of subjects in each analysis population by treatment group will be summarized.
8.1. Screened Analysis Set
The Screened Analysis set will consist of all subjects who signed informed consent and 
undergone screening.
8.2. Randomized Analysis Set
The Randomized Analysis set will consist of all Screened Analysis set subjects who are assigned 
a randomization number in the study. For analysis purposes, subjects will be analyzed according 
to the randomized treatment group. Subjects who are replaced will still be counted as 
randomized subjects.
8.3. Safety Analysis Set
The Safety Analysis set will consist of all randomized subjects who receive at least 1 dose of 
investigational product. In this population, subjects will be summarized based upon the actual 
treatment group.


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8.4. Pharmacokinetics Analysis Set
The Pharmacokinetics Analysis set will consist of all subjects in the Safety Analysis set who 
received at least one dose of investigational product and have at least one on study PK 
measurement. The analysis population for any population PK modeling may be defined 
separately in a population PK data analysis plan.
8.5. Pharmacodynamics Analysis Set
The Pharmacodynamics Analysis set will consist of all subjects in the Safety Analysis set who 
have measurable post-baseline PD data. For analysis purposes, subjects will be analyzed 
according to randomized treatment group. The analysis set will be used for all PD data other than 
MRI and Fibroscan measures. This set will be used for correlation analysis.
8.6. Pharmacodynamics Analysis Set - MRI
The Pharmacodynamics Analysis set – MRI will consist of all subjects in the Safety Analysis set 
who have measurable post-baseline MRI assessment. For analysis purposes, subjects will be 
analyzed according to randomized treatment group. This set will be used for MRI related 
endpoints.
8.7. Pharmacodynamics Analysis Set - Fibroscan
The Pharmacodynamics Analysis set – Fibroscan will consist of all subjects in the Safety 
Analysis set who have measurable post-baseline Fibroscan. For analysis purposes, subjects will 
be analyzed according to randomized treatment group. This set will be used for Fibroscan related 
endpoints.
9. Statistical Methods of Data Analyses
9.1. General Considerations
In general, placebo subjects from each cohort will be pooled for summary.
9.1.1. Statistical Notation and Presentation
All data collected from all subjects in the electronic data capture will be presented in the subject 
level listings. All continuous data will be listed with the same precision as presented in the 
database. Data listings will be sorted by cohort, treatment group, subject ID and time point. 
Missing values will be represented by an empty cell and no imputation will be made. If 
imputation is used for analysis, excluding treatment-emergent and concomitant medication 
determinations, the imputed values will be listed with a derivation type column in the listing 
indicating that origin of the imputed value.
Continuous data will be summarized in tables using number of subjects or sample size (n), mean, 
median, standard deviation (SD), minimum and maximum. For logarithm-transformed data, the 
geometric mean, standard error (SE) of the geometric mean and geometric coefficient of 
variation (CV%) will also be provided.


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Categorical data will be summarized in two ways, by subject and by time point. Subject data will 
be summarized using the count of distinct subjects that fall in the category and the percentage of 
the total number of subjects. Population counts, either the number of subjects or the number of 
assessments at the time point, will be used as the denominator in the calculation of percentages 
unless specified otherwise.
The original units (conventional) and SI units will be presented in listings. In general, 
conventional units will be used for tables and figures, unless otherwise specified. Insulin will use
the conventional unit of uU/mL; estimated glomerular filtration rate will be presented using SI 
units.
Minimum and maximum values will use the precisions of the original value. Means, least 
squares (LS) means, and medians will be rounded to one decimal place greater than the precision 
of the original value up to a max of two decimal places. The limits of confidence intervals, 
standard deviations and standard errors will be rounded to two decimal places greater than the 
precision of the original value. Derived PK and PD data will be presented with three significant 
figures. Percentages will be rounded to the nearest tenth. Two-sided p-values will be presented 
with four decimal places and values less than 0.0001 will be presented as <.0001.
9.1.2. Statistical Modeling Methods
A mixed model repeated measures (MMRM) analysis will be used to analyze the change from 
baseline and/or percent change from baseline in the pharmacodynamic endpoints. The model will 
include baseline as a covariate, treatment group, visit, and the interaction between treatment 
group and visit as factors. The covariance will be unstructured; if the model fails to converge, 
other structures such as compound symmetry will be considered. The LS means and the LS mean 
difference will be presented by visit with their corresponding standard error, p-values and two-
sided 90% and 95% confidence intervals.
If strong evidence exists that the normality assumptions are violated for, but not limited to, 
triglycerides or MRI-PDFF, non-parametric methods will be considered such as the Wilcoxon 
Rank Sum test.
There will be no adjustments for multiplicity in this study.
9.1.3. Analysis Visits and Visit Windowing
In general, all scheduled visits will be used as analysis visit for descriptive analysis tables and 
listings. 
If a scheduled visit is missing, an unscheduled visit may be mapped to take its place if and only 
if it is within the visit window of the missing scheduled visit. The following table describes the 
visit mapping of unscheduled post-baseline visits in the event that a scheduled visit is missing. If 
there are multiple unscheduled visits, the closest one to the projected date will be selected; if 
there are ties, the later measurement will be used.


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Parameter
Analysis Visit/Time Point Window
Labs
Study Day ± 3 days from Target Day
ET visits are mapped to the nearest missing analysis visit,
using the same window, after unscheduled visits are 
considered.
MRI; Fibroscan
Day 50: 36<= Study Day <= 64 (± 14 Days);
Day 92: Study Day >= 78;
ET visits within Study Day < 65 will be mapped to Day 50;
ET visits with Study Day >= 65 will be mapped to Day 92.
Vital Signs
Study Day ± 3 days from Target Day;
ET visits are mapped to the nearest missing analysis visit,
using the same window, after unscheduled visits are 
considered.
Anthropomorphic Measures
Study Day ± 3 days from Target Day (For body weight)
Study Day ± 14 days from Target Day (For waist-hip ratio
related parameters)
ET visits are mapped to the nearest missing analysis visit,
using the same window, after unscheduled visits are 
considered
ECG
Study Day ± 3 days from Target Day ;
ET visits are mapped to the nearest missing analysis visit 
after unscheduled visits are considered using Study Day
PK
Depending on the time point, the window for each time point 
is specified in the PK sampling schedule. If multiple 
unscheduled records are available, the sample collected 
closest to the nominal time will be selected. If the distance is 
the same, the later one will be selected. (Actual time should 
be used for PK analysis)
Measurements from all protocol planned visits will be summarized in descriptive tables and 
figures. For pharmacodynamic endpoints that are analyzed using MMRM analysis, only
measurements taken at weekly analysis visits (eg, Day 1, Day 8, 15, 22, 29, 36, etc) will be 
included in the analysis and corresponding data presentation. Analyses will utilize the 
measurements taken at the scheduled visits whenever they are available. If not available, the 
analysis will use the measurement mapped to that scheduled visit (whichever is the closest to the 
target date, and if there are two measurement before and after the target day with same duration,
use the measurement taken at the later day) If there are duplicate results from the same sample
are available, eg, with the identical sampling date and time, the average value will be used for 
data analysis and both raw data and average value will be listed. EOS/ET visits will be 
summarized in descriptive summary.  ET visits will be eligible to be to mapped to missing 
protocol planned visits, but will not on its own be considered an analysis visit or time point.


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9.1.4. Assessments Below and Above the Limit of Quantification
For summarization of PK concentration values and data analysis, assessments that are below the 
lower limit of quantification (LLOQ) will be set to zero prior to summarization. For PK 
concentration graphs, assessments that are below the lower limit of quantification will be set to 
half of LLOQ prior to graphing. For PK concentration assessments that are above the upper 
limit of quantification (ULOQ), the record will be set to missing prior to summarization or 
graphing. For other exploratory PK analysis such as population PK, assessment that are below 
the LLOQ may be defined differently.
For all other assessments that are below the lower limit of quantification will be imputed to half 
of the LLOQ for summarization; assessments that are above the upper limit of quantification will 
be imputed to the ULOQ for summarization.
For listings, records that are LLOQ or ULOQ will be listed as such together with the imputed 
value in separate columns.
9.2. Pharmacokinetic Analyses
Serum concentration and PK parameter data will analyzed using the Pharmacokinetics Analysis 
Set.
9.2.1. Analysis of Serum Concentrations of BIO89-100
The serum concentration of BIO89-100 over time will be listed and plotted by individual and 
also summarized by treatment group. Serum concentration data will also be presented using 
arithmetic statistics.
9.2.2. Analysis of PK Parameters
The following PK endpoints will be summarized by treatment group:
x
Cmax within a dosing interval (Day 1 and Day 29), the maximum serum concentration 
within each dosing interval.
x
AUC0-tau within a dosing interval (Day 1 and Day 29), derived using linear-log 
trapezoidal rule; assuming linear curve when concentrations are increasing and 
exponential decay when concentrations are decreasing.
x
tmax, the time at which Cmax is observed in reference to the start of the dosing interval 
during dosing interval (Day 1 and Day 29).
x
t1/2, the time at which the concentration is half of the observed maximum concentration 
within the dosing interval during dosing interval (Day 1 and Day 29).
PK parameters will be calculated separately by designate of 89bio.


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9.3. Study Subjects
Subject enrollment by site will be summarized. The number of subjects who were screened, 
randomized, early discontinued from investigational product, early discontinued from study, and 
completers will be summarized.
9.3.1. Subject Disposition
Subject disposition will be presented for the Randomized Analysis set. The number of subjects 
randomized, included in each analysis population, who completed study or had early terminated 
from the study and the reason, who completed IP regimen or discontinued IP regimen 
prematurely and the reason will be summarized by treatment group and overall. The number of 
subjects who were screened will be summarized based on Screened Analysis Set.
Subject disposition will also be summarized by subgroup of biopsy confirmed NASH.
In addition, a listing will be provided for subjects who had been impacted by COVID-19, eg,
study interruptions and the duration of their study interruption, withdrew from investigational 
product, withdrew from study, or had protocol deviation.
9.3.2. Demographics and Baseline Characteristics
Demographics and baseline characteristics data will be summarized descriptively using 
Randomized Analysis Set by treatment group and overall.  Demographics and baseline 
characteristics include but not limited to, age (continuous and FDWHJRULHVWRDQG
years), sex, race and ethnicity, height, body weight, body mass index (continuous and categories: 
<25, 25 to <30, 30 to <35, DQG35 kg/m2), 
, central obesity 
(waist circumference of >102 cm for males, >88 cm for females, or body mass index (BMI) >30 
kg/m2); biopsy confirmed NASH (yes/no);
 
triglycerides (continuous and categories: <150, 150 to <200 DQG200 mg/dL), non-HDL, HDL-
c, LDL-c (continuous and categories: <13130 mg/dL),
, ALT, 
increased ALT ($/78/LQPDOHV$/78/LQIHPDOHV),AST, adiponectin, Pro-C3, 
Pro-C3 subgroup (<14.71, 14.71 ng/mL),
, subjects meeting at least one or two out of the 
following five criteria: baseline ALT >1x ULN, 9&7( N3D(/)3UR-C314.71
ng/mL, FIB-4 score > 1.3. FFA, adipo-IR, hsCRP, MRI-PDFF,
 
, endogenous FGF21, type 2 diabetes mellitus history 
(yes/no), HbA1c, HOMA-IR, fasting plasma glucose,
 
The criteria where the
subjects qualified for the study, eg, 1) NASH with fibrosis (stages F1, F2 or F3) or 2) Central 
obesity with T2DM or 3) Central obesity WITH either increased ALT and/or Fibroscan VCTE 
score 7 KPa per protocol definitions will also be summarized.
Demographics and baseline characteristics summary will be repeated by subgroup of “biopsy 
confirmed NASH with fibrosis status (F1, F2, F3)” (yes/no).


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9.3.3. Biopsy History
The number and percentage of subjects with biopsies within 24 months of screening will be 
summarized by fibrosis stage (F0, F1, F2, F3, F4), NAFLD activity score (continuous with 
range, categorical: score from 0 to 8), steatosis score (continuous with range, categorical), 
ballooning degeneration score (quartiles, categorical), lobular inflammation score (continuous 
with range, categorical). The percentages for this table will be based on the number of subjects 
with biopsies within 24 months.
9.3.4. Medical History
The number and percentage of subjects will be summarized by medical history code (System 
organ class and preferred term) and by treatment group and overall for the Randomized Analysis 
set. Medical history will be coded by Medical Dictionary for Regulatory Activities (MedDRA
version 23.0 – Apr 2020).
9.3.5. Inclusion/Exclusion Criteria
Inclusion and exclusion criteria failures will not be summarized, but will be listed for the 
Screened Analysis set.
9.3.6. Investigational Product Administration
Investigational product administration data such as the number and percentage of subjects who 
were administered full doses of investigational product each time, the number and percentage of 
subjects who were administered at least one partial dose, total number of injections administered
over the study, investigational product exposure and investigational product compliance over the 
entire study will be summarized for the Safety Analysis set. 
All investigational product administration data including injection site, incomplete injections 
their volumes and reason for incompleteness, full or partial doses administered and total volume 
of injections will be listed by visit.
9.3.7. Concomitant Medication
The World Health Organization Drug Dictionary (B3 WHO Drug Global – Mar 2019) will be
used to code the verbatim descriptions of prior and concomitant medications into the Anatomic 
Therapeutic Chemistry (ATC) classification system. Each verbatim name will be classified by 
anatomical main group, ATC level 1, therapeutic subgroup, ATC level 2, pharmacological 
subgroup, ATC level 3, and chemical subgroup, ATC level 4.
The number and percentage of subjects receiving concomitant medications will be summarized 
by treatment group and ATC classification level 2 and level 4 for the Safety Analysis set.
Pre-treatment medications will not be summarized, but will be in listings. 
Baseline medications will also be summarized by ATC classification level 2 and 4.


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9.3.8. Concomitant Procedures
All procedures, pre-treatment and concomitant, will be listed.
9.3.9. Protocol Deviations
Important protocol deviations will be summarized by treatment group for the Randomized 
Analysis set. All protocol deviations will be reviewed by study team to identify notable 
deviations prior to database lock. All protocol deviations will be listed.
9.4. Pharmacodynamic Analyses
Pharmacodynamic analysis will use Pharmacodynamic Analysis Set and will be analyzed based 
on randomized treatment group.
9.4.1. Analyses of Anthropomorphic Measurements
The observed values and the change from baseline and percent change from baseline for body 
weight, BMI, 
will be presented by visit and treatment 
group.
MMRM analysis will be used to analyze the change from baseline and percent change from 
baseline in anthropomorphic measurements. 
Line plots (± SE) of the change from baseline LS mean over time may be presented by treatment 
group.
9.4.2. Analyses of Laboratory Parameters
9.4.2.1.
Lipids
The observed values and the change from baseline and percent change from baseline for 
triglycerides, non-HDL cholesterol, HDL-c, LDL-c,
 will be presented by visit 
and treatment group. Only records where fasting is confirmed will be used for summary and 
analysis.
The following categories will be included in the descriptive summary by visit:
x
Triglycerides <150, 150-<200200 mg/dL
x
LDL-c <130 DQG 130 mg/dL
MMRM analysis will be used to analyze the change from baseline and percent change from 
baseline in lipids.
Non-parametric analysis of the change from baseline and percent change from baseline by visit 
for triglycerides will be compared by treatment group and the p-value from the Wilcoxon Rank 
Sum test will be presented. The Hodges-Lehmann estimate of location shift will be used to 
estimate the median and the exact 90% confidence limits.


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Line plots (± SE) of the change from baseline LS mean over time may be presented by treatment 
group.
In addition, waterfall plots for the individual percent change from baseline to Day 50, change 
from baseline to Day 50, percent change from baseline to Day 92, change from baseline to Day 
92 will be generated.
9.4.2.2.
Metabolic Biomarkers
The observed values and the change from baseline and percent change from baseline for 
,
Adiponectin, HbA1c, HOMA-IR, FFA, Adipo-IR (fasting FFA * fasting insulin), 
,
fasting glucose and fasting insulin (not from OGTT) will be presented by visit and treatment 
group.
HOMA-IR value is calculated by multiplying fasting Glucose (mg/dL) with fasting Insulin
(uIU/ml) and then dividing by 405.
HOMA-IR value (SI units) is calculated by multiplying fasting Glucose (mmol/L) with fasting 
Insulin (uIU/ml) and then dividing by 22.5. 
MMRM analysis will be used to analyze the change from baseline in metabolic biomarkers.
9.4.2.3.
Liver Biomarkers
The observed values and the change from baseline for ALT, AST, 
 
(derived from hyaluronic acid [HA], amino-terminal propeptide of type III procollagen [PIIINP], 
tissue inhibitor of metalloproteinase 1 [TIMP-1]), INR, Pro-C3, total bilirubin (TB), direct 
bilirubin,
will be presented by visit and treatment group.
A listing of only abnormal liver biomarker records will be provided. 
MMRM analysis will be used to analyze the change from baseline in liver biomarkers.
Line plots (± SE) of the change from baseline LS mean over time may be presented by treatment 
group.
9.4.2.3.1.
Sensitivity Analysis for Liver Biomarkers
Subject
had an adverse event that resulted in a spike of ALT and other liver 
biomarkers on Day 92 visit. The event was deemed unlikely related to investigational product.
After receiving medical treatments, these liver biomarkers returned to normal level by End of 
Study visit (Day 113).
In order to estimate the treatment effect assuming the adverse event affecting the liver 
biomarkers did not occur (Hypothetical Strategy), the sensitivity analysis is planned prior to 
study unblinding, and the sensitivity analysis will set this subject’s scheduled Day 92 record as 
missing, and then impute it with the last unscheduled central lab liver biomarkers measured after
Day 92 visit but prior Day 113 visit. Sensitivity analysis using this different estimand will be 
applied to all descriptive, inferential and subgroup analyses summarizing liver biomarkers.


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This sensitivity analysis will only be applied to pharmacodynamic analyses where the PD 
Analysis Set is used. All central lab records will be used for safety analysis purpose without 
impute Day 92 record.
9.4.2.4.
Suspected Drug Induced Liver Injury
Suspected drug induced liver injury will be summarized in two ways.
For subjects with baseline ALT or AST are within normal range, then the proportion of subjects 
meeting the following criteria will be summarized by visit and over the entire study; for entire 
study summary all visits including unscheduled will be considered and the maximum value will 
be selected among all visits:
x
ALT or AST >2x ULN
x
ALT or AST >8x ULN
x
ALT or AST >5x ULN for a duration of at least 2 weeks
x
ALT or AST >3x ULN and (TB >2x ULN or Normalized Ratio (INR) >1.5)
x
ALT or AST >3x ULN (the protocol includes with the criteria the following that will not 
be considered in the summary table: with the appearance of fatigue, nausea, vomiting, 
right upper quadrant pain or tenderness, fever, rash, and/or eosinophilia [> 5%])
For subjects with baseline ALT or AST >ULN, then the proportion of subjects meeting the 
following criteria will be summarized:
x
ALT or AST >2x ULN or bilirubin >1.5x ULN
x
Baseline ALT or AST <2x ULN
o
ALT or AST >5x baseline value
x
%DVHOLQH$/7RU$67[8/1Eut <5x ULN
o
ALT or AST >3x baseline value
x
%DVHOLQH$/7RU$67[8/1
o
ALT or AST >2x baseline value
x
ALT or AST increase >2× baseline value AND the increase is accompanied by a 
concomitant total bilirubin increase to >2× ULN OR the INR concomitantly increases by 
>0.2.
x
ALT or AST increase >2× baseline value (the protocol includes with the criteria the
following that will not be considered in the summary table: in the presence of signs and 
symptom(s) such as fatigue, nausea, vomiting, right upper quadrant pain or tenderness, 
fever, rash, and/or eosinophilia [>5%])
An evaluation of drug induced serious hepatotoxicity (eDISH) plot will be generated.
Any subjects that meet the criteria for suspected DILI will be summarized in a listing.


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9.4.3. Analyses of MRI Measures
The observed values and the change from baseline and percent change from baseline in MRI-
PDFF, 
 will be presented 
by visit and treatment group.
The following criteria will also be summarized at Day 50 and Day 92 based on observed data at 
that visit:
x
Percentage reduction from baseline MRI-3'))and <30%
x
MRI-3'))RU
MMRM analysis will be used to analyze the change from baseline and percent change from 
baseline in MRI-PDFF, abdominal visceral fat, abdominal subcutaneous fat, liver volume.
In addition, a bar chart of LS mean (± SE) percent change from baseline to Day 50, change from 
baseline to Day 50, percent change from baseline to Day 92, change from baseline to Day 92 in 
MRI-PDFF by treatment group.
9.4.3.1.
Responder Analysis
Responder analysis will be performed for MRI-PDFF. The proportion of subjects that reach a 
percentage reduction of 10, 20, 30, 40 and 50 in MRI-PDFF will be summarized based on 
observed data at that visit. The following will be analyzed with a Fisher’s Exact Test.
x
3URSRUWLRQRIVXEMHFWVZKRKDYHSHUFHQWUHGXFWLRQLQ05,-PDFF from baseline to 
Day 92.


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x
3URSRUWLRQRIVXEMHFWVZKRKDYHSHUFHQWUHGXFWLRQLQMRI-PDFF from baseline to 
Day 50.
x
Proportion of subjects who have MRI-PDFF <5% at Day 92.
x
Proportion of subjects who have MRI-PDFF <5% at Day 50.
9.4.3.2.
Supportive Analysis
Supportive analysis of MRI-PDFF will be performed: with sensitivity analysis to assess impact 
of missing data using multiple imputation using a pattern-mixture model (PMM) approach using 
control-based pattern imputation, correlation analysis and non-parametric analysis.
9.4.3.2.1.
Multiple Imputation Using Control Based Pattern-Mixture Model
Multiple imputation will be performed using the placebo group to impute the missing values for 
the BIO89-100 subjects. This imputation will account for the possibility that not all data are 
missing at random. This method of multiple imputation assumes that the missing data post study 
discontinuation for BIO89-100 subjects are missing not at random (MNAR). By using the 
placebo subject data to impute the missing data from the BIO89-100 group, the analysis will 
produce a conservative estimate of the treatment effect of BIO89-100, although not as 
conservative as the worst observed case analysis.
Prior to applying the control-based pattern mixture, the first step will be to impute the non-
monotone missing data assuming the missing at random (MAR) assumption. Only missing data
that are sandwiched by two non-missing records in time will be imputed using the MAR 
assumption. Visit window mapping should also decrease the amount of non-monotone missing 
imputations that need to be done. After the non-monotone missing data patterns are filled in then 
the control-based imputation is applied.
Missing Data Pattern
Baseline
D50
D92
Monotone
X
X
.b
Non-Monotone
X
.a
X
.a is a missing value that will be imputed assuming the MAR assumption because it is sandwiched by two non-
missing records; the earlier records will be imputed first.
.b is a missing value that will be imputed assuming the NMAR assumption because it is monotone missing.
9.4.3.3.
Correlation Analysis
Correlation analysis will be performed for MRI-PDFF and key liver biomarker data using the PD 
analysis set. The pairwise Pearson correlation will be calculated for each combination of MRI-
PDFF, CAP score, VCTE score, ALT, AST, ELF Score, Pro-C3, triglycerides, LDL and HDL
change from baseline within Day 50, within Day 92 and between Day 50 and Day 92 by 
treatment group (subjects on BIO89-100 only) and for pooled placebo.
9.4.3.4.
Non-Parametric Analysis
Non-parametric analysis of the change from baseline and percent change from baseline to Day 
50 and Day 92 for MRI-PDFF will be compared by treatment group and the p-value from the 


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Wilcoxon Rank Sum test will be presented. The Hodges-Lehmann estimate of location shift will 
be used to estimate the median and the exact 90% confidence limits.
 
 
 
 
 
 
 
 
 
 


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9.5. Safety Analyses
Safety analysis will use the Safety Analysis Set and will be analyzed based on actual treatment 
group.
9.5.1. Adverse Events
All adverse events will be coded using the MedDRA (version 23.0 – Apr 2020). 
The incidence and number of events of the following will be summarized by treatment and 
overall:
x
Overall TEAE Summary*
x
TEAEs by System Organ Class (SOC) and Preferred Term (PT)
x
Serious TEAEs by SOC and PT
x
Serious Related TEAEs by SOC and PT 
x
Serious TEAEs Leading to Study Discontinuation by SOC and PT
x
Serious TEAEs Leading to Investigational Product Discontinuation by SOC and PT
x
Related TEAEs by SOC and PT 
x
TEAEs Leading to Study Discontinuation by SOC and PT
x
Fatal TEAE (will be provided in a listing)
x
TEAEs by SOC, PT and Maximum CTCAE Grade
x
TEAEs by SOC, PT and CTCAE Grade
x
TEAEs Leading to Investigational Product Discontinuation by SOC and PT
x
TEAEs by PT in Descending Frequency
x
TEAE by SOC and PT in subgroup of Biopsy confirmed NASH (Yes if F1, F2, F3, else 
No) Yes/No
When calculating incidence; subjects will only be counted once per SOC, PT, or grade.


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When deriving relationship of AEs to study treatment, categories with the relationship of 
“definitely related”, “probably related”, “possibly related” will be considered “related”. If an AE 
has missing relationship, it will be assumed to be related to the investigational product for 
analysis purposes. If an AE has missing CTCAE grade, the missing grade will be imputed as a 
severe grade 3 adverse event. Any TEAE related to VOCID19 will be provided in a listing.
TEAEs by System Organ Class (SOC) and Preferred Term (PT) by subgroup of biopsy 
confirmed NASH (Yes if F1, F2, F3, else No) will be summarized.
*In the Overall TEAE summary, the most severe CTCAE grade will be summarized for each 
subject.
9.5.2. Clinical Laboratory Evaluations
The observed values and the change from baseline for labs (biochemistry, hematology, 
urinalysis) will be summarized by visit and treatment group. Pharmacodynamic endpoints 
summarized via MMRM will not be presented in the safety lab change from baseline tables.
A listing of only abnormal labs (biochemistry, hematology, urinalysis) will be provided.
Shift tables will use the reference range result of low, normal, high to summarize lab 
(biochemistry and hematology) or normal and abnormal for select urinalysis shifts from baseline 
to each post-baseline visit. All labs with a reference range result will be included in shift tables.
The percentage of the shifts will be derived using the number of subjects at the baseline range 
level as the denominator. Pharmacodynamic endpoints summarized via MMRM will be 
presented in the safety lab shift tables.
The observed value and the change from baseline in cortisol will be summarized descriptively in 
the urinalysis tables.
9.5.3. Physical Examination Findings
Physical examination findings will be listed.
9.5.4. Vital Signs
The observed values and the change from baseline for vital signs will be summarized by visit and 
treatment group. On visits where multiple ECGs are taken, only pre-dose will be summarized.
The number of subjects meeting the following criteria will be summarized considering all post-
baseline visits including unscheduled:
x
Absolute value of SBP < 90 mm Hg
x
Absolute value of DBP < 50 mm Hg
x
Pulse rate <50 bpm
x
Pulse rate > 120 bpm
x
0D[LPXPLQFUHDVHIURPEDVHOLQHLQ6%3PP+J
x
0D[LPXPLQFUHDVHIURPEDVHOLQHLQ'%3PP+J


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x
0D[LPXPGHFUHDVHIURPEDVHOLQHLQ6%3PP+J
x
0D[LPXPGHFUHDVHIURPEDVHOLQHLQ'%3PP+J
Shift tables will be used to summarize clinically significant shifts from baseline to each post 
baseline visit in the following criteria:
x
SBP and DBP:
Blood Pressure Category
Systolic mm Hg
Diastolic mm Hg
Hypotension
<90
Or
<60
Normal
<120
And
<80
Elevated
120-129
And
<80
Hypertension Stage 1
130-139
Or
80-89
Hypertension Stage 2

Or

Crisis
>180
And/or
>120
x
HR (bpm): <50; 50-89; 90-99, ı100
9.5.5. 12-Lead Electrocardiograms
The observed values and the change from baseline for 12-lead ECGs parameters will be 
summarized by visit and treatment group. Only visits where ECGs were taken pre-dose will be 
summarized.
For corrected QT interval by Fridericia (QTcF), the number and percentage of subjects that have 
a change from baseline in QTcF >30 to 60msec and >60msec over the entire study considering 
all visits including unscheduled will be summarized.
Overall interpretation (Normal, Abnormal Not Clinically Significant and Abnormal Clinically 
Significant) will also be summarized.
The eCRF and central reader data will be listed in addition to a variable identifying if there is
agreement between the PI and central reader (Y/N).
A shift table will be used to present the shifts from baseline to each visit using the following 
criteria:
x
QTcF <450msec
x
QTcF 450-480msec
x
QTcF 481-500msec
x
QTcF >500msec
x
Not Done/Missing
9.5.6. Columbia-Suicide Severity Rating Scale (C-SSRS)
Suicidal ideation, intensity of ideation, suicidal behavior, and suicidal behavior lethality from C-
SSRS will be listed.


BIO89-100-002
Page 38 of 52
SAP Version 1.0 (Final) – 21AUG2020
9.5.7. Immunogenicity Analyses
Results of immunogenicity assessment will be summarized by displaying the number of subjects 
with positive and negative tests. Antibody titers will be listed. Available binding specificity will 
be listed.
10. Changes to Planned Analysis in the Protocol
There are no changes to the planned analysis in the protocol.
11. Statistical Software
All statistical analyses will be performed using SAS version 9.4 or higher.
12. References
Kahn SE, Lachin JM, Zinman B, et al. Effects of rosiglitazone, glyburide, and metformin on 
beta-cell function and insulin sensitivity in ADOPT. Diabetes. 2011;60(5):1552-1560. doi: 
10.2337/db10-1392.
Matsuda M, DeFronzo RA. Insulin sensitivity indices obtained from oral glucose tolerance 
testing: comparison with the euglycemic insulin clamp. Diabetes Care. 1999; 22(9): 1462-1470.
National Heart, Lung and Blood Institute (2020). Low Blood Pressure. Retrieved May 19, 2020, 
from https://www.nhlbi.nih.gov/health-topics/low-blood-pressure.


BIO89-100-002
Page 39 of 52
SAP Version 1.0 (Final) – 21AUG2020
13. Appendix
Table 2 Schedule of Activities for Cohorts 1-4
Cohorts 1-4 Assessments
Screening Period
Treatment Period
FU Period
Study Visit
Study Day
Informed consent
X
Medical history/demographics
X
Percutaneous liver biopsy 
(optional)b
X
Prior medications
X
Inclusion and exclusion 
criteria
X
X
Complete physical examc
X
X
X
X
Symptom-directed physical 
examd
X
B
P
B
B
B
B
B
Body weighte
X
B
B
B
B
B
B
Waist and hip measurements
X
B
B
X
X
Fibroscanf
X
X
12-lead ECG (single)g
X
B
X
B
B
B
B
X
B
X
X
Urine drug screen and alcohol 
breath test
X
Xh
Clinical laboratory testsi
X
X
B
X
B
B
B
B
X
B
X
X
Cortisol (24 hour urine 
collection)j
X
X
Urinalysis
X
X
X
X
X
HbA1c
X
X
X
X
Serologyk
X
TSH
X


BIO89-100-002
Page 40 of 52
SAP Version 1.0 (Final) – 21AUG2020
Cohorts 1-4 Assessments
Screening Period
Treatment Period
FU Period
Study Visit
Study Day
Pregnancy test in WOCBP 
onlyl
X(S)
X
B
B
B
B
B
B
X
X
Vital signsm
X
X
X P
X X X X
X
X
X
X
P
X
X
X
X
X
X
X
Columbia-Suicide Severity 
Rating Scale (C-SSRS)
X
D50
X
Randomization
X
Investigational product
administrationn
X
X
X
X
X
X
X
PK blood collectiono
X X X X X B
B
B
X
X
X
X
X
B
X
PD and biomarker blood 
sampling:
IGF-1, total
B
B
B
B
X
Adiponectin
B
B
B
X
X
CK-18, ELF panel and Pro-C3
B
B
X
Free fatty acid
B
B
B
X
X
Oral glucose tolerance test 
(OGTT)p
X
X
Insulin (not part of OGTT)
B
B
B
X
HOMA-IR calculation
X
X
X
MRI-PDFF; visceral fat; and 
subcutaneous fatq
X
X
Plasma sample for potential 
future bone biomarkers 
analysisr
X
B at D50
X
X
Pharmacogenomic (DNA) 
blood sampling
X


BIO89-100-002
Page 41 of 52
SAP Version 1.0 (Final) – 21AUG2020
Cohorts 1-4 Assessments
Screening Period
Treatment Period
FU Period
Study Visit
Study Day
Exploratory biomarker 
analysiss
X
B at D50
X
X
Immunogenicityt
B
B
B
B at D50
Xu
X
Endogenous FGF21v
B
Adverse event monitoringw
X
X
X
X X X X X X
X
X
X
X
X
X
X
X
X
X
X
X
Concomitant medication
X
X
X
X X X X X X
X
X
X
X
X
X
X
X
X
X
X
X
Domiciledx
X
X X
X
X
X
Abbreviations: B = predose; D = Day; CK = cytokeratin; DNA = deoxyribonucleic acid; ECG=electrocardiogram; ELF = enhanced liver fibrosis; EOS = end of 
study; ET= early termination; FGF21 = fibroblast growth factor 21; FU = Follow-Up; HbA1c=hemoglobin A1c; IGF-1 = insulin-like growth factor; MRI-PDFF 
= magnetic resonance imaging based proton density fat fraction; P = Pre-discharge; PK = pharmacokinetic; Pro-C3 = N-terminal propeptide of type III collagen; 
S = serum; TSH = thyroid stimulating hormone; WOCBP = women of child-bearing potential.
Table 2 footnotes
All outpatient visits will have a study window of ±1 day.
a.
For any subject who withdraws before completion of the study, an Early Termination (ET) visit will be conducted, if possible; with the same 
assessments as the Day 92 visit.
b.
Optional liver biopsy may be performed for subjects who do not have a medical contraindication to undergoing a liver biopsy, and did not have a liver 
biopsy in the 24 months prior to screening. For subjects who underwent a previous liver biopsy >24 months prior to screening, that would not have 
qualified for the study based on results of the initial biopsy, there should be a sound clinical basis to expect different findings in a repeat biopsy based on 
medical judgment. Any biopsy will need to be approved in advance by Sponsor and only after the subject has met all other inclusion and exclusion 
criteria,
c.
Complete physical exam done at screening to include recording height, weight, and calculating BMI.
d.
Symptom-directed physical exam will be done pre-dose and prior to discharge from the Phase 1 unit on domiciled dosing visits (1st and 5th dose), and 
pre-dose on ambulatory dosing visits. Additional physical examinations will be performed if clinically indicated.
e.
On domiciled and ambulatory visits, body weight will be measured pre-dose.
f.
Fibroscan should be performed during screening, prior to magnetic resonance imaging (MRI) for all subjects.
g.
12-lead safety ECGs will be recorded as single bedside measurements; on domiciled dosing visits (1st and 5th dose), ECG will be measured pre-dose 
and 24 hours post-dose. On ambulatory dosing visits, ECG will be measured pre-dose. Additional ECG may be conducted if clinically indicated.
h.
On Day -1, urine drug screen can be done at a local laboratory, however, a sample should also be collected for central laboratory evaluation.


BIO89-100-002
Page 42 of 52
SAP Version 1.0 (Final) – 21AUG2020
i.
Clinical laboratory tests will include biochemistry, hematology, and coagulation, and FSH for determination of post-menopausal status ; on domiciled 
dosing visits (1st and 5th dose), clinical laboratory will be collected pre-dose and 24 hours post dose; on ambulatory visits, clinical laboratory will be 
collected pre-dose. For all subjects, alanine transaminase (ALT) and aspartate transaminase (AST) will be collected twice during the Screening period 
(at least 2 weeks apart), with the 2nd assessment to be collected at Day -7 to Day -3. A 3rd assessment, if required, will be collected via unscheduled 
visit (refer to Exclusion criterion 3).
j.
Ambulatory 24-hour urine collection for cortisol to be done within 14 days from baseline, on Day 50 and Day 92/ET. It is recommended that subjects 
collect urine for the 24 hours prior to D-7 to-3 visit and bring the container with them to this visit. Alternatively, urine can be collected for the 24 hours 
prior to the randomization visit (D-1) and brought to the site on D-1 (the result is not required for eligibility confirmation). For Day 50 and D92/ET –
subject will start collection 24 hours before coming into the clinic and bring the sample to the visit.
k.
Serology tests will include Hepatitis B surface antigen, Hepatitis C Virus, and Human Immunodeficiency Virus (HIV) 1 and 2 antibodies
l.
Serum urine pregnancy test will be conducted at screening; at all other timepoints urine pregnancy test will be done to guide clinical decisions to dose 
on dosing days. Prior to the 1st dosing, the baseline urine pregnancy test will be performed on D-1 to allow for randomization on that day. If urine test is 
positive, a confirmatory serum pregnancy test will be conducted.
m. Vital signs include supine blood pressure, pulse, body temperature, and respiratory rate; vital signs will be measured pre-dose (prior to scheduled blood 
draws and study intervention administration); 1, 12 and 24 hours post-dose and prior to discharge on domiciled dosing visits (1st and 5th doses); on 
dosing ambulatory visits, vital signs will be measured pre-dose (prior to scheduled blood draws and study intervention administration) and prior to 
discharge; on non-dosing visits, vital signs will be measured prior to scheduled blood draws. Starting from randomization, blood pressure and pulse will 
be measured in duplicate, the first measurement will be taken up to 15 minutes before the indicated time point. Additional vital signs measurement may 
be done if clinically indicated. Subjects must be in a supine or semi-erect/seated position and resting for at least 5 minutes prior to measurements.
n.
Study intervention will be administered SC to the abdomen region by qualified study personnel.
o.
PK blood samples will be collected as shown in Table 3. Additional blood samples for PK analysis may be collected if clinically indicated (e.g., in case 
of SAE). For PK sample collection instruction/procedures, refer to the relevant manual.
p.
Blood samples for OGTT test will be collected under fasting conditions (10 hours) at the following timepoints: 0 minutes (just prior to ingesting 
glucose), 30 minutes, 60 minutes, 90 minutes 120 minutes and 180 minutes. On Day 92/ET the insulin will be captured from the OGTT.
q.
At Screening, MRI-PDFF to be done within 35 days of baseline (Day 1). On Days 50 and 92, MRI-PDFF to be done within ± 2 days of the planned visit 
date. If out of the window, MRI-PDFF should still be performed as close to the target day as possible, but protocol deviation should be recorded.
r.
Samples for carboxy-terminal collagen crosslinks (CTX) and N-terminal propeptide of type 1 collagen (P1NP) will be obtained at the designated 
timepoints for storage and potential future analysis. The D42 sample will be obtained pre-dose.
s.
Samples for RNA as well as plasma and serum samples will be collected for potential future exploratory assessments, to increase understanding of 
BIO89-100 biological activity and to identify potential existing and/or emerging biomarkers.
t.
 
 
 


BIO89-100-002
Page 43 of 52
SAP Version 1.0 (Final) – 21AUG2020
 
u.
Immunogenicity sample on Day 92/ET will only be collected for subjects who early terminate from the study.
v.
Baseline samples of endogenous FGF21 will be analyzed; 
 
w.
The sites may take non-personally identifying photographs of potential injection site reactions (optional)
x.
Subjects in Cohorts 1 to 4 will be domiciled at the clinic from 1 day prior to dosing until 24 hours post the 1st dose and the 5th dose. Other study visits 
will be ambulatory. On Day 8, Day 15 and Day 22 (ambulatory dosing visits), subjects will remain at the site for observation for at least 2 hours post 
dosing.


BIO89-100-002
Page 44 of 52
SAP Version 1.0 (Final) – 21AUG2020
Table 3 PK Sample Collection for Cohorts 1-4 – QW Dosing Interval
Study Day
Dosing 
Day
Ambulatory 
Visit
Hours relative to 1st dose (Day 1)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X


BIO89-100-002
Page 45 of 52
SAP Version 1.0 (Final) – 21AUG2020
Table 4 Schedule of Activities for Cohorts 5 and 6
Cohorts 5-6 Assessments
Screening Period
Treatment Period
FU Period
Study Visit
Study Day
Informed consent
X
Medical history/demographics
X
Percutaneous liver biopsy 
(optional)b
X
Prior medications
X
Inclusion and exclusion 
criteria
X
X
Complete physical examc
X
X
X
X
Symptom-directed physical 
examd
X
B
P
X
B
X
B
B
B
Body weighte
X
B
X
B
X
B
D36
B
B
Waist and hip measurements
X
B
B
X
X
X
Fibroscanf
X
X
X
12-lead ECG (single)g
X
B
X
X
B
B
X
B
X
X
Urine drug screen and alcohol 
breath test
X
Xh
Clinical laboratory testsi
X
X
B
X
X
B
X
B
X
6
X
X
Cortisol (24 hour urine 
collection)j
X
X
Urinalysis
X
X
X
X
X
X
HbA1c
X
X
X
X
Serologyk
X
TSH
X
Pregnancy test in WOCBP 
onlyl
X(S)
X
B
B
B
B
X
X


BIO89-100-002
Page 46 of 52
SAP Version 1.0 (Final) – 21AUG2020
Cohorts 5-6 Assessments
Screening Period
Treatment Period
FU Period
Study Visit
Study Day
Vital signsm
X
X
X P
X X X X
X
X
X
X
P
X
X
X
X
X
X
X
X
Columbia-Suicide Severity 
Rating Scale (C-SSRS)
X
X
X
Randomization
X
Investigational product
administrationn
X
X
X
X
X
PK blood collectiono
X X X X X X
B
X
X
X
X
X
X
X
X
PD and biomarker blood 
sampling:
IGF-1, total
B
X
B
B
X
Adiponectin
B
X
B
X
X
X
CK-18, ELF panel and Pro-C3
B
B
X
X
Free fatty acid
B
X
B
X
X
X
Oral glucose tolerance test 
(OGTT)p
X
X
Insulin (not part of OGTT)
B
X
B
X
X
HOMA-IR calculation
X
X
X
X
MRI-PDFF; visceral fat; and 
subcutaneous fatq
X
X
X
Plasma sample for potential 
future bone biomarkers 
analysisr
X
X
X
X
Pharmacogenomic (DNA) 
blood sampling
X
Exploratory biomarker 
analysiss
X
X
X
X


BIO89-100-002
Page 47 of 52
SAP Version 1.0 (Final) – 21AUG2020
Cohorts 5-6 Assessments
Screening Period
Treatment Period
FU Period
Study Visit
Study Day
Immunogenicityt
B
B
B
X
Xu
X
Endogenous FGF21v
B
Adverse event monitoringw
X
X
X
X X X X X X
X
X
X
X
X
X
X
X
X
X
X
Concomitant medication
X
X
X
X X X X X X
X
X
X
X
X
X
X
X
X
X
X
Domiciledx
X
X X
X
X
X
Abbreviations: B = predose; D = Day; CK = cytokeratin; DNA = deoxyribonucleic acid; ECG=electrocardiogram; ELF = enhanced liver fibrosis; EOS = end of 
study; ET= early termination; FGF21 = fibroblast growth factor 21; FU = Follow-Up; HbA1c=hemoglobin A1c; IGF-1 = insulin-like growth factor; MRI-PDFF 
= magnetic resonance imaging based proton density fat fraction; P = Pre-discharge; PK = pharmacokinetic; Pro-C3 = N-terminal propeptide of type III collagen; 
S = serum; TSH = thyroid stimulating hormone; WOCBP = women of child-bearing potential.
Table 4 footnotes
All outpatient visits will have a study window of ±1 day
* Between Visits 17 to 19, on Days 64 and 78 subjects will be contacted by phone to inquire about AEs and concomitant medications
a.
For any subject who withdraws before completion of the study, an Early Termination (ET) visit will be conducted, if possible; with the same 
assessments as the Day 92 visit.
b.
Optional liver biopsy may be performed for subjects who do not have a medical contraindication to undergoing a liver biopsy, and did not have a liver 
biopsy in the 24 months prior to screening.|For subjects who underwent a previous liver biopsy >24 months prior to screening, that would not have 
qualified for the study based on results of the initial biopsy, there should be a sound clinical basis to expect different findings in a repeat biopsy based on 
medical judgment. Any biopsy will need to be approved in advance by Sponsor and only after the subject has met all other inclusion and exclusion 
criteria.
c.
Complete physical exam done at screening to include recording height, weight, and calculating BMI.
d.
Symptom-directed physical exam will be done pre-dose and prior to discharge from the Phase 1 unit on domiciled dosing visits (1st and 3rd dose), and 
pre-dose on ambulatory dosing visits. Additional physical examinations will also be performed if clinically indicated.
e.
On domiciled and ambulatory visits, body weight will be measured pre-dose.
f.
Fibroscan should be performed during screening, prior to magnetic resonance imaging (MRI) for all subjects.
g.
12-lead safety ECGs will be recorded as single bedside measurements. On domiciled dosing visits (1st and 3rd dose), ECG will be measured pre-dose 
and 24 hours post-dose; on ambulatory dosing visits, ECG will be measured pre-dose. Additional ECG may be conducted if clinically indicated.
h.
On Day -1, urine drug screen can be done at a local laboratory, however, a sample should also be collected for central laboratory evaluation


BIO89-100-002
Page 48 of 52
SAP Version 1.0 (Final) – 21AUG2020
i.
Clinical laboratory tests will include biochemistry, hematology, and coagulation, and FSH for determination of post-menopausal status ; on domiciled 
dosing visits (1st and 3rd dose), clinical laboratory will be collected pre-dose and 24 hours post dose; on ambulatory visits, clinical laboratory will be 
collected pre-dose. For all subjects, alanine transaminase (ALT) and aspartate transaminase (AST) will be collected twice during the Screening period 
(at least 2 weeks apart), with the 2nd assessment to be collected at Day -7 to Day -3. A 3rd assessment, if required, will be collected via unscheduled 
visit (refer to Exclusion criterion 3).
j.
Ambulatory 24 hour urine collection for cortisol to be done within 14 days from baseline, on Day 50 and Day 92/ET. It is recommended that subjects 
collect urine for the 24 hours prior to D-7 to-3 visit and bring the container with them to this visit. Alternatively, urine can be collected for the 24 hours 
prior to the randomization visit (D-1) and brought to the site on D-1 (the result is not required for eligibility confirmation). For Day 50 and D92/ET –
subject will start collection 24 hours before coming into the clinic and bring the sample to the visit.
k.
Serology tests will include Hepatitis B surface antigen, Hepatitis C Virus, and Human Immunodeficiency Virus (HIV) 1 and 2 antibodies
l.
Serum urine pregnancy test will be conducted at screening; at all other timepoints urine pregnancy test will be done to guide clinical decisions to dose 
on dosing days. Prior to the 1st dosing, the baseline urine pregnancy test will be performed on D-1 to allow for randomization on that day. If urine test is 
positive, a confirmatory serum pregnancy test will be conducted.
m. Vital signs include supine blood pressure, pulse, body temperature, and respiratory rate; vital signs will be measured pre-dose (prior to scheduled blood 
draws and study intervention administration); 1, 12 and 24 hours post-dose and prior to discharge on domiciled dosing visits (1st and 3rd doses); on 
dosing ambulatory visits, vital signs will be measured pre-dose (prior to scheduled blood draws and study intervention administration) and prior to 
discharge; on non-dosing visits, vital signs will be measured prior to scheduled blood draws. Starting from randomization, blood pressure and pulse will 
be measured in duplicate, the first measurement will be taken up to 15 minutes before the indicated time point. Additional vital signs measurement may 
be done if clinically indicated. Subjects must be in a supine or semi-erect/seated position and resting for at least 5 minutes prior to measurements.
n.
Study intervention will be administered SC to the abdomen region by qualified study personnel.
o.
PK blood samples will be collected as shown in Table 5. Additional blood samples for PK analysis may be collected if clinically indicated (e.g., in case 
of SAE). For PK sample collection instruction/procedures, refer to the relevant manual.
p.
Blood samples for OGTT test will be collected under fasting conditions (10 hours) at the following timepoints: 0 minutes (just prior to ingesting 
glucose), 30 minutes, 60 minutes, 90 minutes 120 minutes and 180 minutes. . On Day 92/ET the insulin will be captured from the OGTT.
q.
At Screening, MRI-PDFF to be done within 35 days of baseline (Day 1). On Days 50 and 92, MRI-PDFF to be done within ± 2 days of the planned visit 
date. If out of the window, MRI-PDFF should still be performed as close to the target day as possible, but protocol deviation should be recorded.
r.
Samples for carboxy-terminal collagen crosslinks (CTX) and N-terminal propeptide of type 1 collagen (P1NP) will be obtained at the designated 
timepoints for storage and potential future analysis. The D42 sample will be obtained pre-dose
s.
Samples for RNA as well as plasma and serum samples will be collected for potential future exploratory assessments, to increase understanding of 
BIO89-100 biological activity and to identify potential existing and/or emerging biomarkers.
t.
 
 


BIO89-100-002
Page 49 of 52
SAP Version 1.0 (Final) – 21AUG2020
 
u.
Immunogenicity sample on Day 92/ET will only be collected for subjects who early terminate from the study.
v.
Baseline samples of endogenous FGF21 will be analyzed; 
 
w.
The sites may take non-personally identifying photographs of potential injection site reactions (optional)
x.
Subjects in Cohort 5 and 6 will be domiciled at the clinic from 1 day prior to dosing until 24 hours post the 1st dose and 24 hours post the 3rd dose; 
other study visits will be ambulatory. On the Day 15 ambulatory dosing visit, subjects will remain at the study site for observation for at least 2 hours 
post dosing.


BIO89-100-002
Page 50 of 52
SAP Version 1.0 (Final) – 21AUG2020
Table 5 PK Sample Collection for Cohorts 5 and 6 – Q2W Dosing Interval
Study Day
Dosing 
Day
Ambulatory 
Visit
Hours relative to 1st dose (Day 1)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X


BIO89-100-002
SAP Version 1.0 (Final) – 21AUG2020
13.1.
Sample SAS Code
Sample SAS Code for MMRM analysis:
proc mixed data=indata;
class trt visit;
model chg /*or pchg*/ = base trt visit trt*visit/ ddfm=kr s;
repeated visit/type=UN subject=usubjid;
lsmeans trt*visit/pdiff cl alpha=0.1;
run;
Sample SAS Code for MMRM analysis with Fibrosis Status Y/N as factor:
proc mixed data=indata;
class trt visit biopsy_confirmed_NASH_yn;
model chg /*or pchg*/ = base trt visit biopsy_confirmed_NASH_yn trt*visit/ 
ddfm=kr s;
repeated visit/type=UN subject=usubjid;
lsmeans trt*visit/pdiff cl alpha=0.1;
run;
Sample SAS Code for Wilcoxon Rank Sum Non-Parametric analysis:
proc npar1way data=indata wilcoxon alpha=0.1;
class trt;
var chg /*or pchg*/;
exact wilcoxon;
run;
Sample SAS Code for Hodges-Lehman Estimation:
proc npar1way data=indata hl alpha=0.1;
class trt;
var chg /*or pchg*/;
exact hl;
run;
Sample SAS Code for Multiple Imputation Using Control Based Pattern-Mixture Model:
proc mi data = non_mono out=monotone niimpute=100 seed=123;
var trt01pn day50 day92; 
mcmc chain = multiple impute=monotone;
run;
Note: Data needs to be in wide format for the multiple imputation procedure Also, dummy variables are need for 
categorical factors with more than two levels such as TREATMENT.
proc mi data=monotone out=imputed1 nimpute=1 seed = 124;
by _imputation_;
class trt01pn;
var trt01pn day50 day92;
monotone reg(/details);
mnar model (day50 day92/modelobs=( trt01pn =’0’));
run;


BIO89-100-002
SAP Version 1.0 (Final) – 21AUG2020
proc mixed data=imputed;
by _imputation_;
class subject trt visit;
model pchg = base trt visit trt *visit/ddfm=kr;
repeated visit / sub=subject type=un;
lsmeans trt *visit/pdiff cl;
ods output lsmeans=lsmeans diffs=diffs;
run;
proc sort data=lsmeans;
by trt visit _imputation;
run;
proc mianalyze parms=lsmeans;
modeleffects trt * visit;
ods output parameterestimates=lsm;
by trt visit;
run;
proc sort data=diffs;
by visit _imputation;
run;
proc mianalyze parms=diffs;
modeleffects trt * visit;
ods output parameterestimates=dif;
by visit;
run;


DocuSign Envelope ID: A873D7EB-B25E-4EA6-848D-134AB6473798 
SAP Version 1.0 (Final) – 16 DEC 2021 
BIO89-100-002 (Part 2) 
Page 1 of 41 
Statistical Analysis Plan 
Protocol BIO89-100-002 (Part 2, Single Cohort) 
A Randomized, Double-Blind, Placebo-Controlled, Multiple Ascending Dose Study to Evaluate 
the Safety, Tolerability and Pharmacokinetic Properties of BIO89-100 Administered 
Subcutaneously in Subjects with Nonalcoholic Steatohepatitis (NASH) or with Nonalcoholic 
Fatty Liver Disease (NAFLD) and at High Risk of NASH 
Phase 1b/2a 
SAP Version: 1.0 (Final) 
Date: 16 DEC 2021 
Prepared by: 
 Pharmapace 
On Behalf of ProSciento Inc. 
NCT #: NCT04048135


DocuSign Envelope ID: A873D7EB-B25E-4EA6-848D-134AB6473798 
SAP Version 1.0 (Final) – 16 DEC 2021 
 
 
BIO89-100-002 (Part 2) 
Page 2 of 41 
 
 
Statistical Analysis Plan 
Protocol BIO89-100-002 (Part 2, Single Cohort) 
A Randomized, Double-Blind, Placebo-Controlled, Multiple Ascending Dose Study to Evaluate 
the Safety, Tolerability and Pharmacokinetic Properties of BIO89-100 Administered 
Subcutaneously in Subjects with Nonalcoholic Steatohepatitis (NASH) or with Nonalcoholic 
Fatty Liver Disease (NAFLD) and at High Risk of NASH 
Phase 1b/2a 
 
 
SAP Version: 1.0 (Final) 
Date: 16 DEC 2021 
 
This statistical analysis plan has been reviewed and approved by: 
 
 
 
 
-Dec-2021 
 
 
 
 
 
 
 
 
 
 
 -Dec-2021 
 
 
-Dec-2021 
 
 Date: 21
89bio LTD 
 Date: 2
89bio LTD 
 Date: 22
89bio LTD 


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BIO89-100-002 (Part 2) 
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Table of Contents 
1.
Introduction ............................................................................................................................. 8 
2.
Study Objectives ...................................................................................................................... 8 
2.1. Primary Objectives .................................................................................................................. 8 
2.2. Secondary Objectives .............................................................................................................. 8 
3.
Study Overview ....................................................................................................................... 8 
3.1. Study Design ............................................................................................................................ 8 
3.2. Study Activities ....................................................................................................................... 9 
3.3. Randomization Schedule and Blinding Procedures ................................................................. 9 
3.4. Stratification Factors ................................................................................................................ 9 
3.5. Sample Size and Power ........................................................................................................... 9 
3.6. Interim Analysis ...................................................................................................................... 9 
3.7. Safety Monitoring Committee ................................................................................................. 9 
4.
Study Endpoints ..................................................................................................................... 10 
4.1. Primary Endpoints ................................................................................................................. 10 
4.1.1. 
Primary Safety Endpoints ............................................................................................... 10 
4.1.2. 
Primary Efficacy Endpoint ............................................................................................. 10 
4.2. Secondary Endpoints ............................................................................................................. 10 
4.3. Other Safety Endpoints .......................................................................................................... 11 
4.4. Exploratory Endpoints ........................................................................................................... 12 
5.
Definitions ............................................................................................................................. 14 
5.1. Definition of Study Day ......................................................................................................... 14 
5.2. Handling of Missing or Partial Dates .................................................................................... 14 
5.2.1. 
Adverse Event Date Imputations ................................................................................... 14 
5.2.2. 
Medication/Procedure Date Imputations ........................................................................ 15 
5.3. Definition of Study Baseline ................................................................................................. 15 
5.3.1. 
Study Baseline of ALT and AST ................................................................................... 16 
5.3.2. 
Study Baseline for Lipids ............................................................................................... 16 
5.3.3. 
Study Baseline for Vital Signs ....................................................................................... 17 
5.4. Definition of Duration of Exposure ....................................................................................... 17 
5.5. Definition of Age ................................................................................................................... 17 


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5.6. Dose Compliance ................................................................................................................... 17 
5.7. Definition of Pre-treatment Medication and Pre-treatment Procedures ................................ 17 
5.8. Definition of Baseline Medication ........................................................................................ 17 
5.9. Definition of Concomitant Medication and Concomitant Procedures .................................. 17 
5.10. 
Definition of the Open-Label Treatment Group ............................................................ 17 
5.11. 
Definition of Treatment-Emergent Adverse Event ........................................................ 18 
5.13. 
Estimated Glomerular Filtration Rate (CKD-EPI) ......................................................... 18 
5.14. 
Definition of Study Interruption ..................................................................................... 18 
5.15. 
Definition of Histology Outcomes ................................................................................. 19 
6. 
Potential Covariates ............................................................................................................... 19 
7. 
Analysis Sets .......................................................................................................................... 19 
7.1. Full Analysis Set .................................................................................................................... 19 
7.2. Pharmacokinetics Analysis Set .............................................................................................. 19 
7.3. MRI Analysis Set ................................................................................................................... 19 
7.4. Histology Biopsy Analysis Set .............................................................................................. 19 
8. 
Methods of Statistical Analysis ............................................................................................. 19 
8.1. General Considerations .......................................................................................................... 19 
8.1.1. 
Statistical Notation and Presentation .............................................................................. 19 
8.1.2. 
Inferential Statistical Analysis Methods ........................................................................ 20 
8.1.3. 
Analysis Visits and Visit Windowing ............................................................................ 21 
8.1.4. 
Assessments Below and Above the Limit of Quantification ......................................... 22 
8.2. Pharmacokinetic Analyses ..................................................................................................... 22 
8.3. Study Subjects ....................................................................................................................... 22 
8.3.1. 
Subject Disposition ........................................................................................................ 22 
8.3.2. 
Demographics and Baseline Characteristics .................................................................. 23 
8.3.3. 
Baseline Biopsy .............................................................................................................. 23 
8.3.4. 
Medical History .............................................................................................................. 23 
8.3.5. 
Inclusion/Exclusion Criteria ........................................................................................... 23 
8.3.6. 
Investigational Product Administration ......................................................................... 23 
8.3.7. 
Concomitant Medication ................................................................................................ 24 


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8.3.8. 
Concomitant Procedures ................................................................................................ 24 
8.3.9. 
Protocol Deviations ........................................................................................................ 24 
8.4. Pharmacodynamic Analyses .................................................................................................. 24 
8.4.1. 
Analysis of Histology ..................................................................................................... 24 
8.4.1.1. 
Histology Measurements from Central Reader .......................................................... 24 
8.4.2. 
Analyses of Anthropometric Measurements .................................................................. 25 
8.4.3. 
Analyses of Laboratory Parameters ............................................................................... 25 
8.4.3.1. 
Lipids .......................................................................................................................... 25 
8.4.3.2. 
Liver Biomarkers ........................................................................................................ 26 
8.4.3.3. 
Suspected Drug Induced Liver Injury ........................................................................ 26 
8.4.4. 
Analyses of MRI Measures ............................................................................................ 27 
8.5. Safety Analyses ..................................................................................................................... 29 
8.5.1. 
Adverse Events ............................................................................................................... 29 
8.5.2. 
Clinical Laboratory Evaluations ..................................................................................... 29 
8.5.3. 
Physical Examination Findings ...................................................................................... 30 
8.5.4. 
Vital Signs ...................................................................................................................... 30 
8.5.5. 
12-Lead Electrocardiograms .......................................................................................... 31 
8.5.6. 
Columbia-Suicide Severity Rating Scale (C-SSRS) ...................................................... 31 
8.5.7. 
Immunogenicity Analyses .............................................................................................. 31 
9. 
Changes to Planned Analysis in the Protocol ........................................................................ 31 
10. Statistical Software ................................................................................................................ 31 
11. References ............................................................................................................................. 31 
12. Appendix ............................................................................................................................... 32 
12.1. 
Schedule of Activities for Part 2 (Cohort 7) .................................................................. 32 
12.2. 
PK Sample Collection for Part 2 (Cohort 7) .................................................................. 35 
12.3. 
Reference Ranges for Clinical Laboratory Tests ........................................................... 36 


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Abbreviations 
 
ADA 
anti-drug antibodies 
AEs 
adverse events 
AI 
artificial intelligence 
ALT 
alanine transaminase 
ALP 
alkaline phosphatase 
AST 
aspartate transaminase 
ATC 
anatomic therapeutic chemistry 
BMI 
body mass index 
CBC 
complete blood count 
CAP 
controlled attenuation parameter 
CI 
confidence interval 
COVID-19 
Coronavirus disease 2019 
CRO 
clinical research organization 
C-SSRS 
columbia-suicide severity rating scale 
CTCAE 
common terminology criteria for adverse events 
CTX 
carboxy-terminal collagen crosslinks 
DILI 
drug-induced liver injury 
ECG 
electrocardiogram 
eCRF 
electronic Case Report Form 
eDISH 
evaluation of drug induced serious hepatotoxicity 
eGFR 
estimated glomerular filtration rate 
EOS 
end of study 
ET 
early termination 
FGF21 
fibroblast growth factor 21 
FIB-4 
fibrosis-4 score 
FU 
follow-up 
GGT 
gamma-glutamyl transferase 
HbA1c 
hemoglobin A1c 
HDL-c 
high density lipoprotein 
HIV 
human immunodeficiency virus 
ICC 
intraclass correlation coefficient 
INR 
international normalized ratio for prothrombin test 
IP 
investigational product 
LDL-c 
low density lipoprotein 
LLOQ 
lower limit of quantification 
LS Mean 
least-squares mean 
MedDRA 
medical dictionary for regulatory activities 
MMRM 
mixed-model repeated measures 
MRI-PDFF 
magnetic resonance imaging – proton density fat fraction 
n 
sample size 


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NAFLD 
nonalcoholic fatty liver disease 
NAS 
NAFLD activity score 
NASH 
nonalcoholic steatohepatitis 
NASH CRN 
NASH Clinical Research Network 
non-HDL 
non-high density lipoprotein 
P1NP 
n-terminal propeptide of type 1 collagen 
PD 
pharmacodynamics 
PEG 
polyethylene glycol 
PK 
pharmacokinetics 
Pro-C3 
n-terminal propeptide of type iii collagen 
PT 
preferred term 
QTcF 
corrected QT interval by Fridericia 
QW 
weekly 
SAEs 
serious adverse events 
SAP 
statistical analysis plan 
SC 
subcutaneous 
SD 
standard deviation 
SE 
standard error 
SMC 
safety monitoring committee 
SOC 
system organ class 
T2D 
type 2 diabetes mellitus 
TB 
total bilirubin 
TEAE 
treatment-emergent adverse event 
TSH 
thyroid stimulating hormone 
ULN 
upper limit of normal 
ULOQ 
upper limit of quantification 
VCTE 
vibration-controlled transient elastography 
WHO 
world health organization 
WOCBP 
women of child-bearing potential 


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1. Introduction 
This statistical analysis plan (SAP) describes the statistical methods and procedures to be 
implemented in Part 2 of clinical study BIO89-100-002. This statistical analysis plan is based on 
study protocol BIO89-100-002 Version 5.0 dated 11 March 2021. If the data suggest then 
deviations from this plan will be considered. However, any deviations from the SAP must be 
substantiated by sound statistical rationale and documented in the final clinical study report. 
2. Study Objectives 
2.1. Primary Objectives 
The primary objectives of Part 2 of the study are: 
• To evaluate the safety and tolerability of subcutaneous (SC) injections of 27 mg BIO89- 
100, administered weekly for 20 weeks, in subjects with biopsy-proven NASH (NAFLD 
activity score (NAS) ≥4, fibrosis stage F1 with high risk, F2 or F3). 
• To characterize effect of BIO89-100 on liver histology 
2.2. Secondary Objectives 
The secondary objectives of Part 2 of the study are: 
• To characterize biomarkers, pharmacodynamics (PD) profile and biological activity of 
BIO89-100 
• To characterize effect of BIO89-100 on liver histology 
• To characterize BIO89-100 pharmacokinetics (PK) 
3. Study Overview 
3.1. Study Design 
Part 2 of study BIO89-100-002 is an open-label cohort (Cohort 7) in which BIO89-100 will be 
administered SC weekly at a single dose level (27 mg QW) to approximately 20 subjects with 
biopsy-proven NASH (NAS ≥4, fibrosis stage F1 with high risk, F2 or F3), to evaluate effect on 
histological endpoints. 
 
Cohort 
 
Dose Levela 
Frequency and Route of 
Administration 
Number of Subjects 
BIO89-100 
Placebo 
7 
27 mg 
QW, SC to abdomen (2 
injections) Days 1, 8, 15, 22, 29, 
36, 43, 50, 57, 64, 71, 78, 85, 92, 
99, 106, 113, 120, 127, and 134. 
Total doses: 20 
20 
NA 
a The actual doses will be ±5% the mg dose due to technical considerations related to drug withdrawal from the vials 
into the syringes for injection. This difference is considered negligible for subject exposure. 


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The study will include a Screening period, a Treatment period, and a Follow-up period. After 
signing informed consent, subjects will undergo screening assessments to determine eligibility 
over a period of up to 60 days. 
For each subject in Part 2 (Cohort 7), the total duration of study participation will be up to 229 
days (31.5 weeks), excluding the potential study interruption due to the Coronavirus disease 
2019 (COVID-19) pandemic: 
• Screening period: ≤60 days (8.5 weeks) 
• Treatment period: 141 days (20 doses in 19 weeks) 
• Follow-up period 28 days (4 weeks) 
3.2. Study Activities 
Please refer to the appendix for the schedule of activities and PK sample collection schedules: 
• Appendix 12.1 Schedule of Activities for Part 2 (Cohort 7) 
• Appendix 12.2 PK Sample Collection for Part 2 (Cohort 7) 
3.3. Randomization Schedule and Blinding Procedures 
Part 2 will be open-label with all subjects receiving the same dose (i.e., 27 mg SC QW) of 
BIO89-100. 
3.4. Stratification Factors 
There are no stratification factors for Part 2. 
3.5. Sample Size and Power 
For Part 2, no formal power calculations were used to determine the sample size in Cohort 7 and 
the number of subjects was chosen based on clinical experience with similar proof-of-concept 
histology studies and feasibility. This sample size accounts for an anticipated 
 of subjects 
who will not undergo the post-baseline liver biopsy (e.g., 
 to have paired liver biopsies). 
With a sample size of 
, the 95% confidence interval of the histological response rate would 
have a half-width no greater than 
 assuming the histological response 
 The 
actual half-width will depend on the observed histological response rate. 
3.6. Interim Analysis 
No efficacy interim analysis is planned. Sponsor may perform administrative interim analyses to 
support objectives such as study planning and regulatory interactions. 
3.7. Safety Monitoring Committee 
Part 2 will not have a Safety Monitoring Committee (SMC) as described for Part 1 since there 
will be no dose escalation decisions to adjudicate. Part 2 safety data will be reviewed on an 
ongoing basis by the Sponsor and/or clinical research organization (CRO) Medical Monitors. 


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4. Study Endpoints 
4.1. Primary Endpoints 
Part 2 of this study has primary safety and primary efficacy endpoints, which are detailed in 
Sections 4.1.1 and 4.1.2, respectively. 
4.1.1. Primary Safety Endpoints 
The primary safety endpoints are: 
• Subject incidence of treatment-emergent adverse events (TEAEs) 
• Subject incidence of serious TEAEs 
4.1.2. Primary Efficacy Endpoint 
The primary efficacy endpoint is: 
• At least a 2-point improvement in NAS with at least a 1-point improvement in ballooning 
or lobular inflammation, and no worsening of fibrosis 
4.2. Secondary Endpoints 
The secondary PD endpoints are: 
Anthropometric measurements: 
• Change from baseline and percent change from baseline in body weight over time 
Laboratory parameters: 
• Change from baseline and percent change from baseline in triglycerides over time 
o Change from baseline and percent change from baseline in triglycerides over time 
in subjects with baseline triglycerides ≥150 mg/dL 
• Change from baseline and percent change from baseline in non-high density lipoprotein 
(non-HDL) cholesterol over time 
• Change from baseline and percent change from baseline in high density lipoprotein 
(HDL-c) over time 
o Change from baseline and percent change from baseline in HDL-c over time in 
subjects with baseline HDL-c <40 mg/dL for males and <50 mg/dL for females 
• Change from baseline and percent change from baseline in low density lipoprotein 
(LDL-c) over time 
o  Change from baseline and percent change from baseline in LDL-c over time in 
subjects with baseline LDL-c ≥100 mg/dL 
• Change from baseline and percent change from baseline in hemoglobin A1c (HbA1c) 
over time 


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Liver function tests: 
• Change from baseline and percent change from baseline in alanine transaminase (ALT) 
over time 
• Change from baseline and percent change from baseline in aspartate transaminase (AST) 
over time 
• Change from baseline and percent change from baseline in n-terminal propeptide of type 
III collagen (Pro-C3) over time 
Imaging measures: 
• Change from baseline and percent change from baseline in magnetic resonance imaging – 
proton density fat fraction (MRI-PDFF) 
Histology measures: 
• Improvement of fibrosis ≥1 stage without worsening of NASH 
• NASH resolution without worsening of fibrosis 
PK measures: 
• Trough concentration of BIO89-100 
4.3. Other Safety Endpoints 
The other safety endpoints are: 
• Subject incidence of TEAEs by severity 
• Number of subjects who discontinued due to TEAEs 
• Number of subjects who discontinued due to related TEAEs 
• Change from baseline in vital signs 
• Incidence and shifts of clinically significant vital signs 
• Incidence of clinically significant physical examination findings 
• Incidence and shifts of clinically significant electrocardiograms (ECG) 
• Incidence and shifts of clinically significant laboratory abnormalities 
• Change from baseline in safety laboratory evaluations including complete blood count 
(CBC), blood biochemistry, urinalysis and bone biomarkers (CTX and P1NP) 
Immunogenicity measures: 
• Incidence and characteristics of anti-drug antibodies (ADA) after dosing (e.g., titer and/or 
binding specificity, to the fibroblast growth factor 21 (FGF21) and polyethylene glycol 
(PEG) part of BIO89-100 and neutralizing immunogenicity) 
• Impact/correlation of the presence of ADAs on serum BIO89-100 concentrations and 
clinical safety 


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4.4. Exploratory Endpoints 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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5. Definitions 
5.1. Definition of Study Day 
The day subjects receive their first dose of investigational product (IP) will be considered Study 
Day 1. Study day will be defined by the number of days from Study Day 1. If index date is prior 
to Study Day 1, study day will be defined as the index date minus the Study Day 1 date. If index 
date is on or after Study Day 1 date, the study day will be defined as index date minus the Study 
Day 1 date plus 1 day. If a subject did not receive IP, the study day will be set as missing. 
For subjects with study interruptions due to COVID-19, post-interruption study day will be 
calculated as: (Index date) – (the date of Study Day 1) + 1 – (the interruption duration as defined 
in the study interruption CRF). Visits post-interruption due to COVID-19 will use this corrected 
study day for visit windowing. 
5.2. Handling of Missing or Partial Dates 
Date imputation will only be used for determining treatment-emergent status for adverse events 
or identifying concomitant medications. Actual data values as they appear in the database will be 
shown in the subject level listings. 
5.2.1. Adverse Event Date Imputations 
In cases of incomplete dates for adverse events, the missing components will be assumed as the 
most conservative values as possible. The imputation rules described below will attempt to 
conservatively capture AEs with missing onset dates as treatment-emergent, unless information 
of AE stop date/time was prior to first dose date, which would automatically disqualify the 
treatment-emergent status. The original values with no imputation applied will be presented in 
the data listings. 
• If “day” is the only missing field, impute the “day” as the day of Study Day 1 date if the 
month and year of Study Day 1 are equal to the AE start month and year; otherwise, 
impute the “day” as the first day of the AE start month. 
• If “day” and “month” are missing, impute the “day” and “month” as the day and month 
of the first dose date if “year” is the same as the year of Study Day 1; otherwise, impute 
January 1 of the non-missing year. 
• Missing time will not be imputed. If “time” is missing and the start date or imputed start 
date is the same as the first dose date, then the time is assumed to be after the dose time 
so that the event will be classified as treatment-emergent. 
• If the start date is completely missing: 
o and from the end date (either complete or partial date) it can be deduced to be 
prior to the first dose date, then the AE will not be assigned as a TEAE. 
o and from the end date (either complete or partial date) it cannot be deduced to be 
prior to the first dose date, then the AE will be assigned as a TEAE. 
o Start dates that are completely missing will not be imputed. 


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5.2.2. Medication/Procedure Date Imputations 
Imputation of partial start and end dates for medications/procedures is done for the purpose of 
classifying concomitant, pre-treatment, and baseline medications/procedures. The original values 
with no imputation applied will be presented in the data listings. 
For medications/procedures with completely missing start dates, no imputation will be 
performed. 
For medications/procedures with partial start dates: 
•
If “day” is the only missing field, impute the “day” as the first of the month.
•
If “day” and “month” are the missing fields, impute the “day” and “month” to January 1.
•
If only day is missing, and month and year are the same as first dose date then consider as
concomitant. For a medication/procedure that is not checked as ongoing and the start date
is completely missing:
o
and from the end date (either complete, partial or completely missing) it can be
deduced to be prior to the first dose date of IP, then consider as pre-treatment.
o
Otherwise, consider as concomitant.
For medications/procedures with completely missing end dates and ongoing is not checked: 
•
End date will not be imputed, furthermore:
o
If start date is on or after Study Day 1, then consider as concomitant
medication/procedure
o
If start date is prior to Study Day 1, or start date is also completely missing, then
consider as pre-treatment medication/procedure
o
If start date is on Study Day 1, then consider as baseline medication/procedure
For medications/procedures with partial end dates: 
•
If “day” is the only missing field, and if month and year are the same as the start date and
ongoing is not checked, then impute “day” as the start date “day”; otherwise impute as
the last day of the month.
•
If “day” and “month” are the missing fields, if year is the same as the start date and
ongoing is not checked, then impute “day” and month as the start date day and start date
month; otherwise impute as December 31.
For medications/procedures that have both completely missing start dates and end dates, no 
imputation will be made; these medications/procedures will be assumed to be concomitant. 
5.3. Definition of Study Baseline 
For study measurements (safety labs, PD, biomarkers and exploratory labs) except ALT or AST, 
lipids, and vital signs, each subject’s baseline value for that measurement will be defined as the 
latest pre-dose value. This will be determined by chronologically sorting all scheduled and 


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unscheduled values prior to first dose date and time and taking the latest value unless noted 
otherwise. 
Baseline will be presented with each post-baseline change in the summary tables to allow easier 
review. 
5.3.1. Study Baseline of ALT and AST 
For ALT and AST, the baseline value will be defined as the average of all values performed 
during screening and Study Day 1 based on the following table. The baseline for ALT and AST 
are calculated and captured in the electronic Case Report Form (eCRF). 
 
ALT or AST Screening Assessments 
Day 1 ALT 
or AST 
assessment 
Baseline value 
Assessment 1 
Assessment 2 
Assessment 3 
(if applicable) 
Normal 
Normal 
Not applicable 
Any 
Average of Assessment 1, 
Assessment 2 and Day 1 
(3 tests) 
Normal 
Abnormal and 
≤40% increase 
from Assessment 
1 
Not applicable 
Any 
Average of Assessment 1, 
Assessment 2 and Day 1 
(3 tests) 
Normal 
Abnormal and 
>40% increase 
from Assessment 
1 
Normal or 
≤40% increase 
from 
Assessment 1 
Any 
Average of Assessment 1, 
Assessment 2, Assessment 
3 and Day 1 (4 tests) 
Normal 
Abnormal and 
>40% increase 
from Assessment 
1 
Abnormal and 
>40% increase 
from 
Assessment 1 
Not 
applicable, 
subject 
excluded. 
Not applicable, subject 
excluded. 
Abnormal 
≤40% increase 
from Assessment 
1 
Not applicable 
Any 
Average of Assessment 1, 
Assessment 2 and Day 1 
(3 tests) 
Abnormal 
>40% increase 
from Assessment 
1 
≤40% increase 
from 
Assessment 1 
Any 
Average of Assessment 1, 
Assessment 2, Assessment 
3 and Day 1 (4 tests) 
Abnormal 
>40% increase 
from Assessment 
1 
>40% increase 
from 
Assessment 1 
Not 
applicable, 
subject 
excluded. 
Not applicable, subject 
excluded. 
5.3.2. Study Baseline for Lipids 
For lipids, the baseline will be determined as the average of all pre-dose measurements within 30 
days prior to Study Day 1, e.g., measurements taken at -29 ≤ Study Day ≤ 1 (Pre-dose). 


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5.3.3. Study Baseline for Vital Signs 
For vital signs of blood pressure and pulse, measurements will be taken in duplicate starting from 
enrollment. Baseline for these measures should be the average of all individual assessments 
during screening up to Study Day 1 and prior to first dose if subject is dosed. 
5.4. Definition of Duration of Exposure 
The intended duration of exposure for QW dosing will be defined as the date of the last dose of 
IP minus Study Day 1 date plus 8 days. 
5.5. Definition of Age 
Age will be defined as the duration in days of date of signed informed consent or index date 
minus the birthdate divided by 365.25. Index date refers to specific visits if age is needed at 
specific visits (for example, age at specific visits will be used when deriving FIB-4 score or 
eGFR). 
5.6. Dose Compliance 
Dose compliance percentage will be the actual number of received doses divided by the expected 
number of doses to be received up to the end of study (EOS). Partial doses will contribute to the 
actual number of received doses. 
5.7. Definition of Pre-treatment Medication and Pre-treatment Procedures 
Pre-treatment medications are defined as any medication that start and stop being taken prior to 
Study Day 1. 
Pre-treatment procedures are defined as any procedure that were started and completed prior to 
Study Day 1. 
5.8. Definition of Baseline Medication 
Medications that have Study Day 1 within the duration of exposure will be considered baseline 
medications. 
5.9. Definition of Concomitant Medication and Concomitant Procedures 
Concomitant medications are defined as any medication that is taken any day on or after Study 
Day 1. This includes medications that were started prior first dose and that were continued to be 
taken after Study Day 1. 
Concomitant procedures are defined as any procedures that are performed any day on or after 
Study Day 1. This includes procedures that were started prior to first dose and that continued into 
Study Day 1. 
5.10. Definition of the Open-Label Treatment Group 
The definition of the open-label treatment group is BIO89-100 27 mg QW. 


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5.11. Definition of Treatment-Emergent Adverse Event 
Treatment-emergent adverse events will be defined as those occurring at or after the time of the 
first administration of IP, e.g., first dose date and time, through study termination (EOS+28 days 
of reporting window), or existing prior to the time of and worsening after the time of the first 
administration of IP. Adverse events with onset prior to the first administration of IP that end 
before Study Day 1 or do not worsen after Study Day 1 will be classified as pre-treatment. 
 
 
 
 
5.13. Estimated Glomerular Filtration Rate (CKD-EPI) 
The eGFR will be derived using the CKD-EPI equation. 
For Males: 
eGFR[mL/min/1.73m2] 
= 141 
 
∗ min ( Serum Creatinine[mg/dL] 
0.9 
 
, 1) 
−.411 
𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆 𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶[mg/dL] 
∗ max ( 
0.9 
, 1) 
−1.209 
∗ 0.993𝐴𝐴𝐴𝐴𝐴𝐴[𝑦𝑦𝑦𝑦𝑦𝑦𝑦𝑦𝑦𝑦] 
 
For Females: 
∗ 1.159[𝑖𝑖𝑖𝑖 𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏] 
eGFR[mL/min/1.73m2] 
= 141 
Serum Creatinine[mg/dL] 
∗ min ( 
0.7 
 
, 1) 
−.329 
∗ max ( 𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆𝑆 𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶[mg/dL] 
0.7 
, 1) 
−1.209 
∗ 0.993𝐴𝐴𝐴𝐴𝐴𝐴[𝑦𝑦𝑦𝑦𝑦𝑦𝑦𝑦𝑦𝑦] ∗ 1.018 
∗ 1.159[𝑖𝑖𝑖𝑖 𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏] 
5.14. Definition of Study Interruption 
Study interruption will be defined as any interruption lasting ≥2 weeks and allowed up to 4 
weeks (+4 days) that is due to any causes related COVID-19 pandemic and will require extra 
handling when deriving analysis visits. 


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5.15. Definition of Histology Outcomes 
Resolution of NASH includes the total absence of ballooning (score=0) and absent or mild 
inflammation (score 0 to 1). 
Worsening of fibrosis is defined as progression of fibrosis ≥1 stage in NASH CRN fibrosis score. 
Fibrosis improvement is defined as ≥1-stage decrease in NASH CRN fibrosis score. 
Worsening of NASH is defined as increase ≥1 point in NAS for ballooning or inflammation. 
6. Potential Covariates 
There will be no adjustment for covariates. 
7. Analysis Sets 
The number of subjects in each analysis set will be summarized. 
7.1. Full Analysis Set 
The Full Analysis Set will consist of all enrolled subjects in Part 2 who received at least one dose 
of study intervention . The Full Analysis Set will also be used for safety summary and for PD 
endpoints, except for MRI and histology endpoints. 
7.2. Pharmacokinetics Analysis Set 
The PK Analysis Set will consist of all subjects in the Full Analysis Set who have received 
BIO89-100, and have at least one serum BIO89-100 concentration measurement collected with 
full dose, and can adequately characterize the trough serum BIO89-100 concentrations. 
7.3. MRI Analysis Set 
The MRI Analysis Set will consist of all subjects in the Full Analysis Set who have measurable 
post-baseline MRI data. 
7.4. Histology Biopsy Analysis Set 
The Histology Biopsy Analysis Set will consist of all subjects in the Full Analysis Set who have 
both baseline (screening or eligible historical) and post-baseline biopsies. 
8. Methods of Statistical Analysis 
8.1. General Considerations 
In general, scheduled visits from baseline to Day 162 will be summarized. For summaries of 
worst post-baseline or post-baseline min/max, all visits will be considered, including 
unscheduled. 
8.1.1. Statistical Notation and Presentation 
All data collected from all subjects in the electronic data capture will be presented in the subject 
level listings. All continuous data will be listed with the same precision as presented in the 


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database. Data listings will be sorted by subject ID and time point. Missing values will be 
represented by an empty cell and no imputation will be made. If imputation is used for analysis, 
excluding treatment-emergent and concomitant medication determinations, the imputed values 
will be listed with a derivation type column in the listing indicating that origin of the imputed 
value. 
Continuous data will be summarized in tables using number of subjects or sample size (n), mean, 
median, standard deviation (SD), minimum and maximum. 
Categorical data will be summarized in two ways, by subject and by time point. Subject data will 
be summarized using the count of distinct subjects that fall in the category and the percentage of 
the total number of subjects. Population counts, either the number of subjects or the number of 
assessments at the time point, will be used as the denominator in the calculation of percentages 
unless specified otherwise. 
The original units (conventional) and SI units will be presented in listings. In general, 
conventional units will be used for tables and figures, unless otherwise specified. Insulin will use 
the conventional unit of uU/mL; estimated glomerular filtration rate will be presented using SI 
units. 
Minimum and maximum values will use the precisions of the original value. Means, 
least-squares (LS) means, and medians will be rounded to one decimal place greater than the 
precision of the original value up to a max of two decimal places. The limits of confidence 
intervals (CI), standard deviations (SD), and standard errors (SE) will be rounded to two decimal 
places greater than the precision of the original value. Derived PK and PD data (e.g., unit 
conversion) will be presented with three decimal places. Percentages will be rounded to the 
nearest tenth. Two-sided p-values will be presented with four decimal places and values less than 
0.0001 will be presented as <.0001. 
8.1.2. Inferential Statistical Analysis Methods 
A mixed-model repeated measures (MMRM) analysis will be used to analyze the change from 
baseline and/or percent change from baseline for selected PD parameters. The model will include 
baseline value of the dependent variable as a covariate, and subjects as a random effect to 
account for repeated visits. A compound symmetric covariance structure will be used. The LS 
means will be derived from the model with their corresponding SE, p-values and two-sided 95% 
CI. 
If strong evidence exists that the normality assumptions are violated for, but not limited to, 
triglycerides or MRI-PDFF, non-parametric methods will be considered such as the Wilcoxon 
Rank Sum test. 
For proportion of various types or responders, the 95% exact binomial CI will be provided. 


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The Cohan-Kappa statistics, including the Kappa coefficient, 95% CI, and two-sided p-value will 
be applied when assessing the concordant and discordant between two raters, e.g., histology data 
central reader versus PathAI reader. 
There will be no adjustments for multiplicity in this study. 
8.1.3. Analysis Visits and Visit Windowing 
In general, all scheduled visits will be used as analysis visit for descriptive analysis tables and 
listings. 
If a scheduled visit is missing, an unscheduled visit may be mapped to take its place if and only 
if it is within the visit window of the missing scheduled visit. The following table describes the 
visit mapping of unscheduled post-baseline visits in the event that a scheduled visit is missing. If 
there are multiple unscheduled visits, the closest one to the projected date will be selected; if 
there are ties, the later measurement will be used. 
 
Parameter 
Analysis Visit/Time Point Window 
Labs 
Study Day ±3 days from Target Day 
Early termination (ET) visits are mapped to the nearest missing analysis 
visit, using the same window, after unscheduled visits are considered. 
MRI 
Day 85: Study Day >=78; 
ET visits with Study Day >=65 will be mapped to Day 85. 
Day 141: Study Day >=113 
Histology 
Endpoints 
Day 141: Any post-baseline 
Fibroscan 
Day 141: Any post-baseline 
Vital Signs 
Study Day ±3 days from Target Day; 
ET visits are mapped to the nearest missing analysis visit, using the same 
window, after unscheduled visits are considered. 
Anthropometric 
Measures 
Study Day ±3 days from Target Day (For body weight) 
Study Day ±14 days from Target Day (For waist-hip ratio related 
parameters) 
ET visits are mapped to the nearest missing analysis visit, using the same 
window, after unscheduled visits are considered 
ECG 
Study Day ±3 days from Target Day ; 
ET visits are mapped to the nearest missing analysis visit after 
unscheduled visits are considered using Study Day 
PK 
Depending on the time point, the window for each time point is specified 
in the PK sampling schedule. If multiple unscheduled records are 
available, the sample collected closest to the nominal time will be 
selected. If the distance is the same, the later one will be selected. 
 
Measurements from all protocol planned visits will be summarized in descriptive tables and 
figures. For PD endpoints that are analyzed using MMRM analysis, only measurements taken at 


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analysis visits (e.g., Day 1, Day 8, 15, 29, 57, etc.) will be included in the analysis and 
corresponding data presentation. Analyses will utilize the measurements taken at the scheduled 
visits or the unscheduled visit that have been mapped to a scheduled visits, as described in 
Section 8.1.3. If there are duplicate results from the same sample are available, e.g., with the 
identical sampling date and time, the average value will be used for data analysis and both raw 
data and average value will be listed. 
8.1.4. Assessments Below and Above the Limit of Quantification 
For summarization of PK concentration values and data analysis, assessments that are below the 
lower limit of quantification (LLOQ) will be set to zero prior to summarization. For PK 
concentration graphs, assessments that are below the lower limit of quantification will be set to 
half of LLOQ prior to graphing. For PK concentration assessments that are above the upper 
limit of quantification (ULOQ), the record will be set to missing prior to summarization or 
graphing. For other exploratory PK analysis such as population PK, assessment that are below 
the LLOQ may be defined differently. 
For all other assessments that are below the lower limit of quantification will be imputed to half 
of the LLOQ for summarization; assessments that are above the upper limit of quantification will 
be imputed to the ULOQ for summarization. 
For listings, records that are LLOQ or ULOQ will be listed as such together with the imputed 
value in separate columns. 
8.2. Pharmacokinetic Analyses 
Trough serum concentration will be summarized using the PK Analysis Set. 
The trough serum concentration of BIO89-100 over time will be listed and plotted by individual 
subject and also summarized by nominal study visit. Serum concentration data will also be 
presented using descriptive statistics. Mean and individual serum concentration vs. time plots 
will be produced on both the linear/linear scale and on the semi-logarithmic scale. 
8.3. Study Subjects 
Subject enrollment by site will be summarized. The number of subjects who were screened, 
enrolled, early discontinued from IP, early discontinued from study, and completers will be 
summarized. 
8.3.1. Subject Disposition 
Subject disposition will be presented for the Full Analysis Set. The number of subjects enrolled, 
included in each analysis set, who completed study or had early terminated from the study and 
the reason, who completed IP regimen or discontinued IP regimen prematurely and the reason 
will be summarized. The number of subjects who were screened will be summarized. 


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In addition, a listing will be provided for subjects who had been impacted by COVID-19, e.g., 
study interruptions and the duration of their study interruption, withdrew from IP, withdrew from 
study, or had protocol deviation. 
8.3.2. Demographics and Baseline Characteristics 
Demographics and baseline characteristics data will be summarized descriptively using Full 
Analysis Set. Demographics and baseline characteristics include but not limited to, age 
(continuous and categories: <65, 65 to <75 years), sex, race and ethnicity, height, body weight, 
body mass index (continuous and categories: 25 to <30, 30 to <35, and ≥35 kg/m2), 
 
, central obesity (waist circumference of >102 cm for males, >88 cm 
for females, or body mass index (BMI) >30 kg/m2); 
 
, triglycerides (continuous and categories: <150, 150 to <200 and ≥200 
mg/dL), non-HDL, HDL-c, LDL-c (continuous and categories: <130, ≥130 mg/dL), 
 
, ALT, increased ALT (ALT ≥40 U/L in males, ALT ≥30 U/L in females), AST, 
adiponectin, Pro-C3, Pro-C3 subgroup (<14.71, ≥14.71 ng/mL), 
 
; MRI-PDFF, endogenous FGF21, T2D history (yes/no), HbA1c, fasting 
plasma glucose. 
8.3.3. Baseline Biopsy 
The number and percentage of subjects with biopsies within 24 weeks of baseline including 
qualified historical biopsy and on-study biopsy during screening will be summarized by fibrosis 
stage (F2, F3), NAS (continuous with range, categorical: score from 0 to 8), steatosis score 
(continuous with range, categorical), ballooning degeneration score (quartiles, categorical), 
lobular inflammation score (continuous with range, categorical) for the Full Analysis Set. The 
percentages for this table will be based on the number of subjects with biopsies within 24 weeks 
in the Full Analysis Set. 
8.3.4. Medical History 
The number and percentage of subjects will be summarized by medical history code with system 
organ class (SOC) and preferred term (PT) for the Full Analysis Set. Medical history will be 
coded by Medical Dictionary for Regulatory Activities (MedDRA version 23.1 – Sep 2020). 
8.3.5. Inclusion/Exclusion Criteria 
Inclusion and exclusion criteria failures will not be summarized, but will be listed for the Full 
Analysis Set. 
8.3.6. Investigational Product Administration 
IP administration data such as the number and percentage of subjects who were administered full 
doses of IP each time, the number and percentage of subjects who were administered at least one 
partial dose, total number of injections administered over the study, IP exposure, relative dose 
intensity and IP compliance over the entire study will be summarized for the Full Analysis Set. 


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All IP administration data including injection site, volume of incomplete injections -and reason 
for incompleteness, full or partial doses administered and total volume of injections will be listed 
by visit. 
Relative dose intensity is the ratio of actual dose intensity (mg) to planned dose intensity (mg). 
Actual dose intensity (mg) is equal to the cumulative actual dose (mg) divided by 20. Dose 
compliance percentage is defined as the actual number of received doses divided by the expected 
number of doses to be received up to EOS. Partial doses contribute to the actual number of 
received doses. 
8.3.7. Concomitant Medication 
The World Health Organization Drug Dictionary (B3 WHO Drug Global – Mar 2019) will be 
used to code the verbatim descriptions of prior and concomitant medications into the Anatomic 
Therapeutic Chemistry (ATC) classification system. Each verbatim name will be classified by 
anatomical main group, ATC level 1, therapeutic subgroup, ATC level 2, pharmacological 
subgroup, ATC level 3, and chemical subgroup, ATC level 4. 
The number and percentage of subjects receiving concomitant medications will be summarized 
by ATC classification level 2 and level 4 for the Full Analysis set. 
Pre-treatment medications will not be summarized, but will be in listings. 
Baseline medications will also be summarized by ATC classification level 2 and 4. 
8.3.8. Concomitant Procedures 
All procedures, pre-treatment and concomitant, will be listed. 
8.3.9. Protocol Deviations 
Protocol deviations will be summarized for the Full Analysis Set. All protocol deviations will be 
reviewed by study team to identify major deviations prior to database lock. Major protocol 
deviations will be flagged and summarized as well. All protocol deviations will be listed. 
8.4. Pharmacodynamic Analyses 
PD analysis will use Full Analysis Set, MRI Analysis Set, and Histology Biopsy Analysis Set. 
8.4.1. Analysis of Histology 
8.4.1.1. 
Histology Measurements from Central Reader 
Endpoints of different histological responses will be summarized with the point estimates and 
95% CIs of the proportions of subjects who meet the response criteria (defined in Sections 4.1.2, 
4.2, and 4.4) at Day 141 using the Histology Biopsy Analysis Set. 
Baseline, change from baseline in individual score including NASH CRN fibrosis score, ISHAK 
fibrosis score, NAS score, steatosis grade, lobular inflammation grade, and ballooning grade will 
be summarized using the Histology Biopsy Analysis Set and by subgroups. The proportions of 


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subjects who met ≥1-point improvement in steatosis, ≥1-point improvement in ballooning, 
≥1-point improvement in inflammation will be summarized in tables and plotted in bar charts. 
Shift of fibrosis stage from baseline to Day 141 in CRN and ISHAK fibrosis scores will be 
summarized. Shift of individual NAS components from baseline to Day 141 will be summarized. 
Bar charts of CRN and ISHAK fibrosis scores with baseline and Day 141 side by side may be 
presented. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
8.4.2. Analyses of Anthropometric Measurements 
The observed values and the change from baseline and percent change from baseline for body 
weight, BMI, 
 will be presented by visit. 
MMRM analysis will be performed on weight. Line plots (±SE) of the change from baseline 
mean over time may be presented. 
8.4.3. Analyses of Laboratory Parameters 
8.4.3.1. 
Lipids 
The observed values and the change from baseline and percent change from baseline for 
triglycerides, non-HDL cholesterol, HDL-c, LDL-c, 
 will be presented by visit. 


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MMRM analysis will be performed on all lipids parameters. Only records where fasting is 
confirmed will be used for summary and analysis. 
The following categories will be included in the descriptive summary by visit: 
• Triglycerides <150, 150 - <200, and ≥200 mg/dL 
• LDL-c <130 and ≥130 mg/dL 
Additional shift tables for lipids based on the following cut points will be provided for baseline 
versus post-baseline values, by visit: 
• Triglycerides <150, 150 - <200, and ≥200 mg/dL 
• Triglycerides <150, ≥150 mg/dL 
• LDL-c <130 and ≥130 mg/dL 
• LDL-c <100 and ≥100 mg/dL 
• HDL-c <40 and ≥40 mg/dL for males, and <50 and ≥50 mg/dL for females 
Line plots (±SE) of the change from baseline mean over time may be presented. 
In addition, waterfall plots for the individual percent change from baseline to Day 85, change 
from baseline to Day 85, percent change from baseline to Day 113, change from baseline to Day 
113 will be generated. 
8.4.3.2. 
Liver Biomarkers 
The observed values and the change from baseline for ALT, AST, Pro-C3, and 
 will be 
presented by visit. MMRM analysis will be performed on all four endpoints. 
A listing of only abnormal liver biomarker records will be provided. 
Line plots (±SE) of the change from baseline mean over time may be presented. 
8.4.3.3. 
Suspected Drug Induced Liver Injury 
Suspected drug induced liver injury will be summarized in two ways. 
For subjects with baseline ALT or AST are within normal range, then the proportion of subjects 
meeting the following criteria will be summarized by visit and over the entire study; for entire 
study summary all visits including unscheduled will be considered and the maximum value will 
be selected among all visits: 
• ALT or AST >2x ULN 
• ALT or AST >8x ULN 
• ALT or AST >5x ULN for a duration of at least 2 weeks 
• ALT or AST >3x ULN and (total bilirubin (TB) >2x ULN or international normalized 
ratio (INR) >1.5) 


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• ALT or AST >3x ULN (the protocol includes with the criteria the following that will not 
be considered in the summary table: with the appearance of fatigue, nausea, vomiting, 
right upper quadrant pain or tenderness, fever, rash, and/or eosinophilia [>5%]) 
For subjects with baseline ALT or AST >ULN, then the proportion of subjects meeting the 
following criteria will be summarized: 
• ALT or AST >2x ULN or bilirubin >1.5x ULN 
• Baseline ALT or AST <2x ULN 
o ALT or AST >5x baseline value 
• Baseline ALT or AST ≥2x ULN but <5x ULN 
o ALT or AST >3x baseline value 
• Baseline ALT or AST ≥5x ULN 
o ALT or AST >2x baseline value 
• ALT or AST increase >2x baseline value AND the increase is accompanied by a 
concomitant total bilirubin increase to >2x ULN OR the INR concomitantly increases by 
>0.2. 
• ALT or AST increase >2x baseline value (the protocol includes with the criteria the 
following that will not be considered in the summary table: in the presence of signs and 
symptom(s) such as fatigue, nausea, vomiting, right upper quadrant pain or tenderness, 
fever, rash, and/or eosinophilia [>5%]) 
An evaluation of drug induced serious hepatotoxicity (eDISH) plot will be generated. If there are 
subjects with Hy’s law then these subjects’ data will be listed. 
Any subjects that meet the criteria for suspected drug-induced liver injury (DILI) will be 
summarized in a listing. 
8.4.4. Analyses of MRI Measures 
The observed values and the change from baseline and percent change from baseline in 
MRI-PDFF will be presented by visit. MMRM analysis will also be performed. 
Number of subjects with MRI-PDFF ≥5% or <5% will be summarized at Day 85 and Day 141 
based on observed data at each visit. The responder analysis will be performed for MRI-PDFF 
using the thresholds of reaching ≥30, ≥50, and ≥70 reduction in MRI-PDFF based on observed 
data at each visit. Ninety-five percent confidence intervals will be provided for the proportion of 
responders. 
In addition, a bar chart will be generated including mean (±SE) percent change from baseline to 
Day 85, change from baseline to Day 85, percent change from baseline to Day 141, and change 
from baseline to Day 141 in MRI-PDFF. 


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8.5. Safety Analyses 
Safety analysis will use the Full Analysis Set. 
8.5.1. Adverse Events 
All adverse events will be coded using the MedDRA (version 23.1 – Sep 2020). 
The incidence and number of events of the following will be summarized: 
• Overall TEAE Summary* 
• TEAEs by System Organ Class (SOC) and Preferred Term (PT) 
• Serious TEAEs by SOC and PT 
• Serious Related TEAEs by SOC and PT 
• Serious TEAEs Leading to Study Discontinuation by SOC and PT 
• Serious TEAEs Leading to IP Discontinuation by SOC and PT 
• Related TEAEs by SOC and PT 
• TEAEs Leading to Study Discontinuation by SOC and PT 
• Fatal TEAE (will be provided in a listing) 
• TEAEs by SOC, PT and Maximum CTCAE Grade 
• TEAEs by SOC, PT and CTCAE Grade 
• TEAEs Leading to IP Discontinuation by SOC and PT 
• TEAEs by PT in Descending Frequency 
When calculating incidence; subjects will only be counted once per SOC, PT, or grade. 
When deriving relationship of AEs to study treatment, categories with the relationship of 
“definitely related”, “probably related”, and “possibly related” will be considered “related”.  If 
an AE has missing relationship, it will be assumed to be related to the IP for analysis purposes. If 
an AE has missing CTCAE grade, the missing grade will not be imputed. A separate category for 
“missing” will be included in the table. Any TEAE related to COVID19 will be provided in a 
listing. 
*In the Overall TEAE summary, the most severe CTCAE grade will be summarized for each 
subject. 
8.5.2. Clinical Laboratory Evaluations 
The observed values and the change from baseline for labs (biochemistry, hematology, and 
urinalysis) will be summarized by visit. Pharmacodynamic endpoints summarized via descriptive 
efficacy summaries will not be presented in the safety lab change from baseline tables. 
MMRM analysis will be performed on HbA1c and adiponectin. 
A listing of only abnormal labs (biochemistry, hematology, and urinalysis) will be provided. 


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Shift tables will use the reference range result of low, normal, high to summarize lab 
(biochemistry and hematology) or normal and abnormal for select urinalysis shifts from baseline 
to each post-baseline visit. All labs with a reference range result will be included in shift tables. 
The percentage of the shifts will be derived using the number of subjects at the baseline range 
level as the denominator. The normal and abnormal reference range is included in Appendix 12.3 
in the appendix section. 
8.5.3. Physical Examination Findings 
Physical examination findings will be listed. 
8.5.4. Vital Signs 
The observed values and the change from baseline for vital signs will be summarized by visit. 
The number of subjects meeting the following criteria will be summarized considering all post- 
baseline visits including unscheduled: 
• Absolute value of SBP <90 mm Hg 
• Absolute value of DBP <50 mm Hg 
• Pulse rate <50 bpm 
• Pulse rate >120 bpm 
• Maximum increase from baseline in SBP ≥30 mm Hg 
• Maximum increase from baseline in DBP ≥20 mm Hg 
• Maximum decrease from baseline in SBP ≥30 mm Hg 
• Maximum decrease from baseline in DBP ≥20 mm Hg 
Shift tables will be used to summarize clinically significant shifts from baseline to each post 
baseline visit in the following criteria: 
• SBP and DBP: 
 
Blood Pressure Category 
Systolic mm Hg 
Diastolic mm Hg 
Hypotension 
<90 
Or 
<60 
Normal 
<120 
And 
<80 
Elevated 
120-129 
And 
<80 
Hypertension Stage 1 
130-139 
Or 
80-89 
Hypertension Stage 2 
≥140 
Or 
≥90 
Crisis 
>180 
And/or 
>120 
• HR (bpm): <50; 50-89; 90-99, ≥100 
For the assignment of blood pressure category, since they are not mutually exclusive, the 
assignment priority is as follows: Crisis, Hypertension Stage 2, Hypertension Stage 1, Elevated, 
Normal and then Hypotension. 


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8.5.5. 12-Lead Electrocardiograms 
The observed values and the change from baseline for 12-lead ECGs parameters will be 
summarized by visit. Only visits where ECGs were taken pre-dose will be summarized. 
For corrected QT interval by Fridericia (QTcF), the number and percentage of subjects that have 
a change from baseline in QTcF >30 to 60 msec and >60 msec over the entire study considering 
all visits including unscheduled will be summarized. 
Overall interpretation (Normal, Abnormal Not Clinically Significant and Abnormal Clinically 
Significant) will also be summarized. 
The eCRF and central reader data will be listed in addition to a variable identifying if there is 
agreement between the PI and central reader (Y/N). 
A shift table will be used to present the shifts from baseline to each post-baseline visit using the 
following criteria: 
•
QTcF <450 msec
•
QTcF 450-480 msec
•
QTcF 481-500 msec
•
QTcF >500 msec
•
Not Done/Missing
8.5.6. Columbia-Suicide Severity Rating Scale (C-SSRS) 
Suicidal ideation, intensity of ideation, suicidal behavior, and suicidal behavior lethality from 
C-SSRS will be listed.
8.5.7. Immunogenicity Analyses
Immunogenicity assessment will utilize a tiered approach (i.e., screen, confirm, titer, binding 
specificity and neutralizing activity). Results of immunogenicity assessment will be summarized 
by displaying the number and proportion of subjects with positive and negative tests. Antibody 
titers will be listed and summarized (i.e., n, mean, SD, median, min, max) by study visit. 
Available binding specificity and neutralizing immunogenicity results will be listed. 
9. Changes to Planned Analysis in the Protocol
There are no changes to the planned analysis in the protocol.
10. Statistical Software
All statistical analyses will be performed using SAS version 9.4 or higher.
11. References
National Heart, Lung and Blood Institute (2020). Low Blood Pressure. Retrieved May 19, 2020, 
from https://www.nhlbi.nih.gov/health-topics/low-blood-pressure. 


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12. Appendix
12.1. Schedule of Activities for Part 2 (Cohort 7)
Cohorts 7 Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit 
Study Day 
(mandatory in clinic visits) 
Informed consent 
X 
Medical history/demographics 
X 
Percutaneous liver biopsy 
(optional)b 
X 
X 
Histology machine read 
(PathAI) 
X 
X 
Prior medications 
X 
Inclusion and exclusion criteria 
X 
B 
Complete physical examc 
X 
X 
X 
Symptom-directed physical 
examd 
B 
Body weight 
X 
B 
X 
X 
Waist and hip circumference 
X 
B 
X 
Lifestyle counseling 
X 
X 
X 
Fibroscane 
X 
X 
12-lead ECG (single)f 
X 
B 
X 
X 
Urine drug screen and alcohol 
breath test 
X 
Bg
Clinical laboratory testsh 
X 
X 
B 
X 
X 
Urinalysis 
X 
B 
X 
X 
HbA1c 
X 
B 
X 
X 
Serologyi 
X 
TSH 
X 
Pregnancy test in WOCBP onlyj 
X(S) 
B 
X 
X 
Vital signsk
X 
B 
X 
X 
Columbia-Suicide Severity 
Rating Scale (C-SSRS) 
X 
Study intervention in-clinic 
(mandatory) administrationl 
X 
Home or in-clinic study 
intervention administrationm 
PK blood collectionn
B 
X 
Pro-C3 
B 
X 
Adiponectin 
B 
X 
MRI-PDFFo
X 
X 
Bone biomarkers analysisp 
B 
X 
X 


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Cohorts 7 Assessments 
Screening Period 
Treatment Period 
FU Period 
Study Visit 
Study Day 
(mandatory in clinic visits) 
Immunogenicityq 
B 
X 
X 
Endogenous FGF21r 
B 
Adverse event monitorings 
X 
X 
X 
X 
X 
Concomitant medication 
X 
X 
X 
X 
X 
Abbreviations: B = predose; D = Day; ECG=electrocardiogram; EOS = end of study; ET= early termination; FGF21 
= fibroblast growth factor 21; FU = Follow-Up; HbA1c=hemoglobin A1c; MRI-PDFF = magnetic resonance 
imaging based proton density fat fraction; PK = pharmacokinetic; Pro-C3 = N‐terminal propeptide of type III 
collagen; S = serum; TSH = thyroid stimulating hormone; WOCBP = women of child-bearing potential. 
Footnotes 
All ambulatory clinic visits will have a study window of ±1 day. Window for MRI-PDFF will be ±7 days. The first 
liver biopsy will be done during the screening period. The second liver biopsy will be performed within 14 days 
after the last dose of study intervention. Liver biopsies (first and second) will be done after MRI-PDFF. 
a. 
For any subject who withdraws before completion of the study, an Early Termination (ET) visit will be 
conducted, if possible; with the same assessments as the Day 141 visit. 
b. A liver biopsy with NASH that meets study inclusion criteria, performed within 24 weeks from baseline 
visit with the sample deemed interpretable by the central reader is acceptable instead of the baseline liver 
biopsy. A second liver biopsy will be done within 14 days of the last dose of study intervention. Liver 
biopsies (first and second) will be done after MRI-PDFF. If out of a window, a liver biopsy should still be 
performed as close to the target date as possible, but a protocol deviation should be recorded. 
c. 
Complete physical exam done at screening to include recording height, weight, and calculating BMI. 
d. Physical examinations will be performed if clinically indicated. 
e. 
Fibroscan should be performed during screening, prior to magnetic resonance imaging (MRI) for all 
subjects. Fibroscan will be repeated on D141/ET. 
f. 
12-lead safety ECGs will be recorded as single bedside measurements. Additional ECG may be conducted 
if clinically indicated. 
g. On Day 1, urine drug screen can be done at a local laboratory. 
h. Clinical laboratory tests (performed under fasting conditions, ≥10 hours) will include biochemistry, 
hematology, and coagulation, and FSH for determination of post-menopausal status. For all subjects, 
alanine transaminase (ALT) and aspartate transaminase (AST) will be collected twice during the Screening 
period (at least 2 weeks apart), with the 2nd assessment to be collected at Day -7 to Day -3. A 3rd 
assessment, if required, will be collected via unscheduled visit (refer to Exclusion criterion 3). 
i. 
Serology tests will include Hepatitis B surface antigen, Hepatitis C Virus, and Human Immunodeficiency 
Virus (HIV) 1 and 2 antibodies. 
j. 
Serum urine pregnancy test will be conducted at screening; at all other timepoints urine pregnancy test will 
be done to guide clinical decisions to dose on dosing days. On dosing days in the clinic, a urine pregnancy 
test will be obtained locally; in addition, a urine sample will be sent to the central lab. If urine test is 
positive, a confirmatory serum pregnancy test will be conducted. 
k. Vital signs include blood pressure, pulse, body temperature, and respiratory rate; vital signs will be 
measured pre-dose (prior to scheduled blood draws and study intervention administration. Blood pressure 
and pulse will be measured in duplicate, the first measurement will be taken up to 15 minutes before the 
indicated timepoint. Additional vital signs measurement may be done if clinically indicated. Subjects must 
be in a supine or semi-erect/seated position and resting for at least 5 minutes prior to measurements. 


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l. 
Study intervention will be administered SC to the abdomen region by qualified study personnel or by the 
subject (under supervision) at ambulatory clinic visit and will be administered at home by the subject at the 
rest of the planned dosing days. Subjects will be observed for 15 minutes after each dose. 
m. At the designated timepoints, study intervention will be administered at the subject’s home. If home 
administration is not feasible, not desired by the subject, or deemed inappropriate option by the investigator 
for any reason, in-clinic study intervention administration may take place at some or all of these timepoints. 
n. PK blood samples will be collected at the specified timepoints. Additional blood samples for PK analysis 
may be collected if clinically indicated (e.g., in case of SAE). For PK sample collection 
instruction/procedures, refer to the relevant manual. 
o. At Screening, MRI-PDFF to be done within 35 days of baseline (Day 1). On Days 85 and 141, MRI-PDFF 
to be done within ±7 days of the planned visit date. If out of the window, MRI-PDFF should still be 
performed as close to the target day as possible, but protocol deviation should be recorded. 
p. Samples for carboxy-terminal collagen crosslinks (CTX), N-terminal propeptide of type 1 collagen (P1NP) 
and osteocalcin will be obtained at the designated timepoints. 
q. Subjects who test positive for neutralizing antibodies to BIO89-100 at the EOS visit (or at the ET visit if 
withdrawn from study) 
 
 
 
 
 
 
 
 
 
 
r. 
Baseline samples of endogenous FGF21 will be obtained in all subjects;
 
 
 
s. The sites may take non-personally identifying photographs of potential injection site reactions (optional). 


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12.2. PK Sample Collection for Part 2 (Cohort 7) 
Study Day 
Dosing 
Day 
Ambulatory 
Visit 
Hours relative to 1st dose (Day 1) 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 


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12.3. Reference Ranges for Clinical Laboratory Tests 


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