Clinical trial NCT04541186
Conditions: Severe Hypertriglyceridemia
Keywords: Hypertriglyceridemia Dyslipidemia Lipid disorder FGF21
Sponsor: 89bio, Inc.
Start date: 2020-09-01; Completion date: 2022-05-31
Enrollment: 86 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
This study is designed to assess the efficacy, safety, and tolerability of different doses and dose regimens (once weekly \[QW\] or every 2 weeks \[Q2W\]), subcutaneous (SC) dosing of BIO89-100 (pegozafermin) compared to placebo in participants with severe hypertriglyceridemia (SHTG).

Detailed description:

Arms: arm 1: Participants received pegozafermin 9 mg QW as an SC injection for 8 weeks in the Main Study cohort. arm 2: Participants received pegozafermin 18 mg QW as an SC injection for 8 weeks in the Main Study cohort. arm 3: Participants received pegozafermin 27 mg QW as an SC injection for 8 weeks in the Main Study cohort or Fibrate Expansion cohort. Main Study and Fibrate Expansion pegozafermin 27 mg QW groups were pooled together due to low sample size in the expansion cohort. arm 4: Participants received pegozafermin 36 mg Q2W as an SC injection for 8 weeks in the Main Study cohort. arm 5: Matching placebo was injected at matching frequency per assigned cohort for 8 weeks in the Main Study cohort or Fibrate Expansion cohort. Main Study and Fibrate Expansion placebo groups were pooled together due to low sample size in the expansion cohort.
Interventions: intervention 1: Pegozafermin intervention 2: Placebo
Intervention descriptions: intervention 1: Subcutaneous injection intervention 2: Matching placebo

Protocol documents:
Prot_000.pdf: 
CLINICAL STUDY PROTOCOL 
Title Page 
Protocol Title: 
A Phase 2, Randomized, Double-Blind, Placebo-Controlled Study 
to Explore the Efficacy and Safety of BIO89-100 in Subjects with 
Severe Hypertriglyceridemia 
Protocol Number:  
BIO89-100-221 
Compound Number: 
BIO89-100 
Study Phase:  
Phase 2 
Sponsor Name: 
89bio, Inc. 
Legal Registered 
Address: 
142 Sansome Street, Second Floor. San Francisco, CA 94104, USA 
Regulatory Agency 
Identifier Number(s): 
IND: 146569 
Sponsor Representative: 
 
Approval Date:  
03 May 2021 
Version: 
3.0 
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 Code of 
Federal Regulations (CFR), and European Union Directives and Regulations (as applicable in the 
region of the study); national country legislation; and 89bio’s Standard Operating Procedures.  
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, Inc. The recipient agrees that no information contained herein may be published or 
disclosed without the written approval of 89bio, Inc. If there is any actual or suspected breach of 
this obligation, the recipient agrees to promptly notify 89bio, Inc. 
©2020 89bio, Inc. All rights reserved 
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TABLE OF CONTENTS 
SIGNATURE PAGE ....................................................................................................................... 2 
SPONSOR SIGNATORY: .............................................................................................................. 2 
PRINCIPAL INVESTIGATOR SIGNATURE PAGE ................................................................... 3 
PROTOCOL AMENDMENT SUMMARY OF CHANGES .......................................................... 4 
TABLE OF CONTENTS ................................................................................................................ 7 
1.
PROTOCOL SUMMARY ......................................................................................... 11 
1.1. 
Synopsis ...................................................................................................................... 11 
OBJECTIVES AND ENDPOINTS FOR MAIN AND FIBRATE EXPANSION COHORTS .... 12 
Overall Design ............................................................................................................................... 14 
NUMBER OF SUBJECTS: ........................................................................................................... 15 
INTERVENTION GROUPS AND DURATION: ........................................................................ 15 
1.2. 
Schedule of Assessments (SoA) ................................................................................. 19 
2.
INTRODUCTION ...................................................................................................... 25 
2.1. 
Study Rationale .......................................................................................................... 25 
2.2. 
Background ................................................................................................................. 26 
2.2.1. 
Severe Hypertriglyceridemia ...................................................................................... 26 
2.2.2. 
FGF21 ......................................................................................................................... 27 
2.2.3. 
BIO89-100 .................................................................................................................. 28 
2.3. 
Benefit/Risk Assessment ............................................................................................ 29 
2.3.1. 
Risk Assessment ......................................................................................................... 29 
2.3.2. 
Benefit Assessment .................................................................................................... 31 
2.3.3. 
Overall Benefit-Risk Assessment ............................................................................... 31 
3.
OBJECTIVES AND ENDPOINTS ............................................................................ 33 
4.
STUDY DESIGN ....................................................................................................... 36 
4.1. 
Overall Design ............................................................................................................ 36 
4.2. 
Scientific Rationale for Study Design ........................................................................ 37 
4.3. 
Justification for Dose .................................................................................................. 37 
4.4. 
End of Study Definition .............................................................................................. 38 
5.
STUDY POPULATION ............................................................................................. 39 
5.1. 
Inclusion Criteria ........................................................................................................ 39 
5.2. 
Exclusion Criteria ....................................................................................................... 40 
5.3. 
Lifestyle Considerations ............................................................................................. 43 
5.3.1. 
Meals and Dietary Restrictions .................................................................................. 43 
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5.3.2. 
Alcohol ....................................................................................................................... 43 
5.3.3. 
Activity ....................................................................................................................... 43 
5.4. 
Screen Failures ........................................................................................................... 43 
6.
INVESTIGATIONAL PRODUCT ............................................................................ 44 
6.1. 
Investigational Product(s) Administered .................................................................... 44 
6.2. 
Administration Instructions ........................................................................................ 44 
6.3. 
Preparation/Handling/Storage/Accountability ........................................................... 45 
6.4. 
Measures to Minimize Bias: Randomization and Blinding ........................................ 45 
6.5. 
Emergency Unblinding ............................................................................................... 46 
6.6. 
Investigational Product Compliance ........................................................................... 46 
6.7. 
Concomitant Therapy ................................................................................................. 46 
6.7.1. 
Prohibited Medications/Therapies .............................................................................. 47 
6.7.2. 
Blinded Alerts and Rescue Medications/Therapies .................................................... 47 
6.8. 
Stopping Rules ............................................................................................................ 48 
6.8.1. 
Monitoring and Discontinuation for Suspected Drug-induced Liver Injury (DILI) 
Criteria ........................................................................................................................ 48 
7.
INTERRUPTION OR DISCONTINUATION OF INVESTIGATIONAL
PRODUCT AND SUBJECT DISCONTINUATION/WITHDRAWAL ................... 50 
7.1. 
COVID-19-Related Risk Mitigation and Interruption of Investigational Product 
Administration ............................................................................................................ 50 
7.2. 
Discontinuation of Investigational Product ................................................................ 51 
7.3. 
Subject Discontinuation/Withdrawal from the Study ................................................. 51 
7.4. 
Lost to Follow-up ....................................................................................................... 52 
8.
STUDY ASSESSMENTS AND PROCEDURES ..................................................... 53 
8.1. 
Efficacy Assessments ................................................................................................. 53 
8.2. 
Safety Assessments .................................................................................................... 53 
8.2.1. 
Physical Examinations ................................................................................................ 54 
8.2.2. 
Vital Signs .................................................................................................................. 54 
8.2.3. 
Electrocardiograms ..................................................................................................... 54 
8.2.4. 
Clinical Safety Laboratory Assessments .................................................................... 54 
8.3. 
Adverse Events and Serious Adverse Events ............................................................. 55 
8.3.1. 
Time Period and Frequency for Collecting AE and SAE Information ...................... 55 
8.3.2. 
Method of Detecting AEs and SAEs .......................................................................... 56 
8.3.3. 
Follow-up of AEs and SAEs ...................................................................................... 56 
8.3.4. 
Regulatory Reporting Requirements for SAEs .......................................................... 56 
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8.3.5. 
Pregnancy ................................................................................................................... 56 
8.4. 
Treatment of Overdose ............................................................................................... 57 
8.5. 
Pharmacokinetics ........................................................................................................ 57 
8.6. 
Pharmacodynamics and Biomarkers .......................................................................... 59 
8.6.1. 
Pharmacodynamics ..................................................................................................... 59 
8.6.3. 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction .............. 60 
8.6.4. 
Exploratory Analyses ................................................................................................. 61 
9.
STATISTICAL CONSIDERATIONS ....................................................................... 62 
9.1. 
Statistical Hypotheses ................................................................................................. 62 
9.2. 
Sample Size Determination ........................................................................................ 62 
9.3. 
Populations for Analyses ............................................................................................ 63 
9.4. 
Statistical Analyses ..................................................................................................... 63 
9.4.1. 
General Considerations .............................................................................................. 63 
9.4.2. 
Primary Endpoint(s) ................................................................................................... 64 
9.4.3. 
Secondary Endpoint(s) ............................................................................................... 64 
9.4.4. 
Exploratory Endpoints ................................................................................................ 65 
9.4.5. 
Safety Analyses .......................................................................................................... 65 
9.5. 
Interim Analyses ......................................................................................................... 65 
9.6. 
Data Monitoring Committee ....................................................................................... 65 
10.
SUPPORTING DOCUMENTATION AND OPERATIONAL
CONSIDERATIONS ................................................................................................. 66 
10.1. 
Appendix 1: Regulatory, Ethical, and Study Oversight Considerations .................... 66 
10.1.1. 
Regulatory and Ethical Considerations ...................................................................... 66 
10.1.2. 
Financial Disclosure ................................................................................................... 66 
10.1.3. 
Informed Consent Process .......................................................................................... 66 
10.1.4. 
Data Protection ........................................................................................................... 67 
10.1.5. 
Dissemination of Clinical Study Data ........................................................................ 67 
10.1.6. 
Data Quality Assurance .............................................................................................. 67 
10.1.7. 
Source Documents ...................................................................................................... 68 
10.1.8. 
Study and Site Start and Closure ................................................................................ 68 
10.1.9. 
Publication Policy ....................................................................................................... 69 
10.2. 
Appendix 2: Clinical Laboratory Tests ...................................................................... 70 
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10.3. 
Appendix 3: Adverse Events: Definitions and Procedures for Recording, 
Evaluating, Follow-up, and Reporting ....................................................................... 72 
10.3.1. 
Definition of AE ......................................................................................................... 72 
10.3.2. 
Definition of Suspected and Unsuspected Adverse Reaction .................................... 73 
10.3.3. 
Definition of Events to Monitor ................................................................................. 73 
10.3.4. 
Definition of SAE ....................................................................................................... 73 
10.3.5. 
Recording and Follow-Up of AE and/or SAE ............................................................ 74 
10.3.6. 
Reporting of SAEs ...................................................................................................... 76 
10.4. 
Appendix 4: Contraceptive Guidance and Collection of Pregnancy Information ...... 77 
10.5. 
Appendix 5: Abbreviations ......................................................................................... 80 
11.
REFERENCES ........................................................................................................... 85 
LIST OF TABLES 
Table 1: 
Intervention Groups Across Main and Fibrate Expansion Cohorts ............................ 16 
Table 2: 
Schedule of Assessments ............................................................................................ 19 
Table 3: 
Discontinuation Criteria in Subjects with Abnormal Baseline ALT or AST Values . 49 
Table 4: 
Adverse Event Reporting Periods .............................................................................. 55 
Table 5: 
Intensive PK Scheme for Sample Collection ............................................................. 58 
Table 6: 
Protocol-Required Laboratory Assessments .............................................................. 70 
LIST OF FIGURES 
Figure 1: 
Study Schema: Main Cohort ...................................................................................... 18 
Figure 2: 
Study Schema: Fibrate Expansion Cohort .................................................................. 18 
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1.
PROTOCOL SUMMARY
1.1. 
Synopsis
Protocol Title: A Phase 2, Randomized, Double-Blind, Placebo-Controlled Study to Explore the 
Efficacy and Safety of BIO89-100 in Subjects with Severe Hypertriglyceridemia (SHTG) 
Rationale: BIO89-100 is a glycoPEGylated analog of fibroblast growth factor 21 (FGF21), 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 (Li, et al. 2015; 
Ogawa, et al. 2007). Administration of exogenous FGF21 is being explored as a method to treat 
hypertriglyceridemia which is associated with increased risk of acute pancreatitis (AP) and 
cardiovascular disease (CVD). Severe hypertriglyceridemia (SHTG), defined as fasting triglyceride 
(TG) levels of greater than or equal to 500 mg/dL, is associated with an increased risk of AP, 
accounting for up to 10% of all AP episodes (de Pretis, 2018), and an increased risk of non-alcoholic 
fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and CVD (Grundy, et al. 2019). 
Clinical practice guidelines from both the American College of Cardiology/American Heart 
Association and from the American Association of Clinical Endocrinologists/American College of 
Endocrinology emphasize the importance of reducing TG in patients with levels above 500 mg/dL. 
Current treatments for hypertriglyceridemia include fibrates, fish oils, niacin, and statins. While these 
treatments can reduce TG levels, not all patients can reduce TG levels to below 500 mg/dL, signifying 
the need for additional therapies. 
Several non-clinical and clinical studies have shown that administration of various FGF21 analogs had 
beneficial effects on serum lipid and insulin-resistance as well as on liver fat (Zhang, et al. 2014, 
Gaich, et al. 2013, Sanyal, et al. 2018). A pharmacodynamic (PD) effect of BIO89-100 on TGs has 
been demonstrated, with significant reductions in TG 
 
 
 
 
In Part 1 of the Phase 1b/2a study BIO89-100-002, subjects with NASH or NAFLD and high risk for 
NASH (hypertriglyceridemia was not an enrollment criterion) who received BIO89-100 at 27 mg QW 
for 12 weeks showed a significant beneficial effect on the lipid profile, evaluated as percent change at 
Week 13 (Day 92) (-28% decrease in TG, -16% reduction in non-HDL cholesterol and LDL 
cholesterol, p<0.05 for all comparisons). 
The current data suggest that BIO89-100 achieves potent and rapid reductions in TG levels through 
pathways mediated by FGF21, offering novel mechanisms of action. Severe hypertriglyceridemia is 
associated with serious outcomes including risk of significant morbidity and mortality, and BIO89-100 
has the potential for development as an important new therapy. 
This Phase 2 study is designed to assess the efficacy, safety, and tolerability of different doses and 
dose regimens (QW or every 2 weeks [Q2W]), subcutaneous (SC) dosing of BIO89-100 compared to 
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placebo in subjects with SHTG. All subjects may be on a background therapy of prescribed fish oils or 
statins but not on niacin, proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors, or any 
supplements, including non-prescribed fish oils, used to alter lipid metabolism. Subjects who are not 
taking prescription therapy to lower serum TG may also be enrolled. 
 Subjects in the main cohort will not be on concurrent fibrate therapy. In contrast, subjects in the 
fibrate expansion cohort will remain on their current stable dose of fibrate therapy.  
The objectives and endpoints are the same for the main cohort and the fibrate expansion cohort. 
Objectives and Endpoints for Main and Fibrate Expansion Cohorts 
Objectives 
Endpoints 
Primary 
•
To determine the effect of
BIO89-100 on serum TG
levels in subjects with
SHTG (TG ≥500 mg/dL)
•
Percentage change in serum TG from baseline to Week 8
Secondary 
•
To determine the effect of
BIO89-100 on selected
serum lipids and
lipoproteins
•
Achieve TG <500 mg/dL at Week 8
•
Percentage change in very low-density lipoprotein
cholesterol (VLDL-C), low-density lipoprotein
cholesterol (LDL-C), non-high-density lipoprotein
cholesterol (non-HDL-C), high-density lipoprotein
cholesterol (HDL-C), very low-density lipoprotein
triglycerides (VLDL-TG), apolipoprotein B100 (ApoB),
remnant lipoprotein cholesterol (RLP-C) from baseline
to Week 8
•
To determine the effect of
BIO89-100 on high-
sensitivity C-reactive
protein (hsCRP)
•
Percentage change in hsCRP from baseline to Week 8
•
To determine the effect of
BIO89-100 on metabolic
markers
•
Percentage change in fasting plasma glucose,
adiponectin, and body weight from baseline to Week 8
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Objectives 
Endpoints 
• To characterize BIO89-100 
pharmacokinetics (PK) 
• Serum BIO89-100 concentration  
• PK parameters in Intensive PK subgroup  
o Maximal observed serum concentrations (Cmax) 
within a dosing interval 
o Area under the serum drug concentration by time 
curve within a dosing interval (AUC0-tau) 
o Time to achieve Cmax (tmax) 
o Terminal elimination half-life (t½) 
Note: Additional PK parameters may be calculated if also 
deemed appropriate. 
• To characterize BIO89-100 
PD profile as assessed by 
MRI-PDFF 
• Percentage change and change from baseline in hepatic 
steatosis using MRI-PDFF  
Safety Endpoints 
 
• To evaluate the safety and 
tolerability of BIO89-100 
• Frequency of treatment-emergent adverse events 
(TEAEs) 
• Frequency of treatment-emergent serious adverse events 
(TESAEs) 
• Number of subjects who discontinue due to TEAEs 
• 
 
• To determine the effect of 
BIO89-100 on liver 
function biomarkers 
• Change in alanine aminotransferase (ALT) and aspartate 
aminotransferase (AST) from baseline to Week 8 
• Other safety assessments 
• Incidence and shifts of clinically significant vital signs, 
physical examination findings, electrocardiogram (ECG) 
data, and laboratory abnormalities; safety laboratory 
evaluations include complete blood count (CBC), blood 
biochemistry, 24-hour urine cortisol, and urinalysis 
• 
 
 
 
 
 
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Overall Design 
Study BIO89-100-221 is a randomized, double-blind, placebo-controlled, Phase 2 study to evaluate the 
efficacy, safety, tolerability, PK and PD profiles, and 
 of BIO89-100 administered SC 
for 8 weeks. The main cohort will include ~90 subjects with SHTG without concurrent fibrate therapy 
and will consist of 5 treatment groups to compare 4 dose levels/regimens of BIO89-100 versus 
placebo. The fibrate expansion cohort will include ~36 subjects with SHTG on stable background 
fibrate therapy and with a baseline MRI-PDFF ≥ 6.0% and will consist of 2 treatment groups 
comparing one dose regimen of BIO89-100 versus placebo. Investigators whose sites are obtaining 
MRIs should assess if subjects on background fibrate therapy are appropriate for the main cohort 
(acceptable to washout fibrate therapy) or for the fibrate expansion cohort. For participation in the 
fibrate expansion cohort, investigators should first discuss the potential subject with the Medical 
Monitor. 
The study includes lifestyle stabilization, TG qualification, treatment, and follow-up periods. 
After signing the informed consent form and confirmation of general eligibility, subjects will undergo 
further screening assessments to confirm eligibility over a period of ~6 to 9 weeks. Generally, eligible 
subjects will enter a 4- or 6-week lifestyle stabilization period (maximum of 6 weeks for subjects 
stopping lipid-altering therapy, minimum of 4 weeks for subjects already on stable, protocol-accepted 
lipid-altering therapy), followed by a ~2-week TG qualification period with qualifying TG 
measurements 
. If mean TG levels are 
not within the inclusion range, an additional week will be allowed for an additional measurement  
  
Individuals who do not meet the criteria for participation in this study (screen failures other than for 
TGs) may be re-screened, at least 7 days later, if the reason for screen failure is deemed temporary and 
is resolved prior to re-screening (e.g., respiratory infection). Re-screened subjects will sign a new 
consent and be assigned a new ID. Subjects can only re-screen once. Re-screening subjects who have 
maintained the lifestyle stabilization period may be allowed to forgo repeated tests 
 
at the discretion of the Medical Monitor. Re-screening after COVID-19 infection requires Medical 
Monitor approval.  
Subjects fulfilling all of the qualifying criteria and none of the exclusionary criteria will be randomized 
1:1:1:1:1 for the main cohort 
 and enter a double-blind, 8-week treatment period during which 
they will be dosed with 1 of 4 doses of BIO89-100 (9 mg QW, 18 mg QW, 27 mg QW, or 36 mg 
Q2W) or placebo.  
Subjects will be enrolled in the fibrate expansion cohort with randomization 1:1 
 and similarly 
enter a double-blind, 8-week treatment period during which subjects will be dosed with BIO89-100 
27 mg or placebo QW.  
Randomization in the main cohort will be stratified by TG <750 mg/dL or ≥750 mg/dL (8.47 mmol/L) 
and by whether or not the subject is receiving background therapy (e.g., prescription fish oil and/or 
statins). Randomization in the fibrate expansion cohort will be stratified by TG <750 mg/dL or 
≥750 mg/dL (8.47 mmol/L) only.   
Treatment with investigational product (IP) may be given at home by a home health provider (HHP) or 
at the Investigator site, QW, SC into the abdomen. To maintain the blind, all subjects will receive 2 SC 
injections at study 
. Subjects randomized to 36 mg Q2W will receive placebo 
at visits between BIO89-100 Q2W administrations. Intensive PK is planned to be assessed in a subset 
of subjects (optional participation to include no more than 45 subjects). Treatment assignment and 
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results of the post-baseline lipid panels (TG, 
 LDL-C, HDL-C, non-HDL-C, 
VLDL-C, VLDL-TG, 
 ApoB, 
 RPL-C, 
), post-baseline MRI-
PDFF, 
, and PK data will be blinded to subjects, sites, study team and 89bio.  
The efficacy, safety, and tolerability of the treatments will be monitored throughout the study as 
described in the objectives (Section 3).  
Subjects who terminate treatment early will be requested to have an EOT 
 
 and an EOS visit 4 weeks later. All subjects who complete the treatment 
period will enter a 4-week follow-up period. 
Disclosure Statement: This is a parallel treatment study in which subjects, Principal Investigators, 
study site personnel, and 89bio will be blinded to treatment during the double-blind treatment period. 
Number of Subjects: 
Approximately 90 subjects are planned to be enrolled in the main cohort, with 18 subjects in each of 
the BIO89-100 dose groups (9 mg QW, 18 mg QW, 27 mg QW, or 36 mg Q2W) and in the placebo 
group. The study aims to enroll ~33% of subjects (~30 subjects total) in the main cohort with a 
 
Subjects who terminate early in either cohort may be replaced at Sponsor’s discretion.  
Intervention Groups and Duration: 
The main cohort will consist of 5 treatment groups (details shown in Table 1), with subjects 
randomized 1:1:1:1:1 to receive 1 of 4 doses of BIO89-100 (9 mg QW, 18 mg QW, 27 mg QW, or 
36 mg Q2W) or placebo. The fibrate expansion cohort will consist of 2 treatment groups randomized 
1:1 to receive BIO89-100 27 mg or placebo QW. Randomization in the main cohort will be stratified 
by TG <750 mg/dL or ≥750 mg/dL (8.47 mmol/L) and by whether or not they are receiving 
background lipid-modifying therapy, as defined by prescription fish oil and/or statins. Randomization 
in the fibrate expansion cohort will be stratified by TG <750 mg/dL or ≥750 mg/dL (8.47 mmol/L) 
only. In both the main and fibrate expansion cohorts, the total duration of treatment will be 8 weeks, 
followed by a 4-week follow-up period. 
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Table 1: 
Intervention Groups Across Main and Fibrate Expansion Cohorts 
Treatment 
Group 
Dose 
Levela 
Frequency and Route of Administration 
Number of 
Subjects 
1 
9 mg 
QW, SC to abdomen 
 
Main cohort only 
18 
2 
18 mg 
QW, SC to abdomen 
 
Main cohort only 
18 
3 
27 mg 
QW, SC to abdomen 
 
Note:  18 subjects in the main cohort and 18 subjects in the 
fibrate expansion cohort 
36 
4 
36 mg 
Q2W, 
BIO89-100: SC to abdomen 
 
Subjects in the 36 mg treatment group will receive placebo at 
visits between BIO89-100 Q2W administrations to maintain 
the blind. 
Placebo: 
 
Main cohort only 
18 
5 
Placebo 
QW, SC to abdomen 
 
Note: 18 subjects in the main cohort and 18 subjects in the 
fibrate expansion cohort 
36 
Abbreviations: SC = subcutaneous; QW = weekly; Q2W = every 2 weeks 
a The actual doses will be ±5% of 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 includes lifestyle stabilization, TG qualification, treatment, and follow-up periods. 
Screening period: 
~6 to 9 weeks 
Lifestyle stabilization 
period: 
~4 weeks (minimum) to ~6 weeks (maximum) 
(~6 weeks for subjects stopping lipid-altering therapy, ~4 weeks for 
subjects already on stable, protocol accepted, lipid-altering therapy) 
TG qualification 
period 
~2 weeks to ~3 weeks 
Treatment period 
8 weeks 
Follow-up period 
4 weeks 
Data monitoring committee: No data monitoring committee is planned for this study. 
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Figure 1: 
Study Schema: Main Cohort 
 
Figure 2: 
Study Schema: Fibrate Expansion Cohort 
 
Abbreviations: EOT = end of treatment, EOS =end of study, N = number of subjects, QW = once weekly. Q2W = every 2 weeks, TG = triglyceride, V = visit, 
wk = week. 
Lifestyle stabilization
TG Qualification
BIO89-100 27 mg QW (N=18)
BIO89-100 18 mg QW (N=18)
BIO89-100 9 mg QW (N=18)
Placebo QW(N=18)
Treatment Period
8 Week
BIO89-100 36 mg Q2W (N=18)
4 (or 6) weeks
2 (or 3) weeks
EOT
Follow-up Period
4 Week
Randomization (1:1:1:1:1)
EOS
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1.2. 
Schedule of Assessments (SoA) 
Table 2: 
Schedule of Assessments 
Schedule of Assessments 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Informed consent 
x 
 
 
 
 
 
 
 
 
 
 
 
 
Inclusion/exclusion 
x 
x 
x 
x 
 
 
 
 
 
 
 
 
 
Randomization 
 
 
 
x 
 
 
 
 
 
 
 
 
 
Demography, medical history 
x 
 
 
 
 
 
 
 
 
 
 
 
 
Physical examination 
x 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
x 
x 
Body weight 
x 
x 
x 
x 
 
 
 
x 
 
 
 
x 
x 
Heightd 
x 
 
 
 
 
 
 
 
 
 
 
 
 
BMI (derived) 
x 
 
 
 
 
 
 
 
 
 
 
x 
 
Vital signs (BPe, heart ratee, RR) 
and body temperature 
x 
x 
 
x 
 
 
 
x 
 
 
 
x 
x 
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Schedule of Assessments 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Hematology, blood chemistry and 
coagulation panel 
xf 
 
xf 
xf 
 
 
 
xf 
 
 
 
xf 
xf 
Urinalysis 
x 
 
 
x 
 
 
 
x 
 
 
 
x 
x 
Thyroid panel (TSH, free T4, and 
free T3) 
x 
 
x 
 
 
 
 
 
 
 
 
x 
x 
12-lead ECG  
x 
 
 
x 
 
 
 
x 
 
 
 
x 
x 
History of drug and/or alcohol 
abuse, urine drug test, and 
alcohol consumption test 
xg 
 
xg,h 
xh 
 
 
 
xh 
xh 
 
xh 
xh 
xh 
Serum lipids, lipoproteins and 
apolipoproteins (TG, 
, VLDL-
C, VLDL-TG, LDL-C, HDL-C, 
non-HDL-C, RLP-C, 
, 
ApoB, 
, 
) 
 
x 
xi 
x 
 
 
 
x 
xi 
 
xi 
x 
x 
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Schedule of Assessments 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Inflammatory biomarker 
(hsCRP) 
 
 
 
x 
 
 
 
x 
 
 
 
x 
x 
 and serum adiponectin, total 
 
 
 
x 
 
 
 
x 
 
 
 
x 
x 
Intensive PK (optional)k 
 
 
 
x 
x 
x 
x 
x 
x 
x 
x 
x 
 
Trough PK (for all subjects)k  
 
 
 
x 
x 
x 
 
x 
 
 
 
x  
 
HIV, HCV and HBV Serologyl  
 
x 
 
 
 
 
 
 
 
 
 
 
 
FSH if required to determine 
menopausal status 
 
x 
 
 
 
 
 
 
 
 
 
 
 
Serum pregnancy test (WOCBP)  
 
x 
 
 
 
 
 
 
 
 
 
 
x 
Urine pregnancy test (WOCBP) 
 
 
x 
x 
 
 
 
x 
 
 
 
x 
 
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Schedule of Assessments 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Jug dispensed for 24-hour urine 
collection for cortisol and 
creatininem 
 
 
x 
 
 
 
 
 
 
 
x 
 
 
Archived serum and plasma 
samplesn 
 
 
 
x 
 
 
 
x 
 
 
 
x 
 
Investigational product dosingp 
 
 
 
x 
x 
x 
x 
x 
x 
x 
x 
  
 
AE and SAEs including 
hypersensitivity-related AEsq 
x 
x 
 
x 
 
 
 
x 
 
 
 
x 
x 
Concomitant medication 
x 
x 
 
x 
 
 
 
x 
 
 
 
x 
x 
Review of lifestyle and alcohol 
user 
x 
x 
x 
x 
 
 
 
x 
x 
 
x 
x 
x 
MRI-PDFF s 
 
 
 
x 
 
 
 
 
 
 
 
x 
 
FGF21 
 
 
 
x 
 
 
 
 
 
 
 
 
 
Shaded columns indicate visits that can be conducted by home health providers. 
Abbreviations: AE = adverse event; 
; ALT = alanine aminotransferase; ALP = alkaline phosphatase; 
; 
ApoB = apolipoprotein B100; 
; aPTT = activated partial thromboplastin time; AST = aspartate aminotransferase; 
 
; BP = blood pressure; BMI = body mass index; BUN = Blood urea nitrogen; CK = creatine kinase; CKD-EPI = chronic kidney 
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disease epidemiology collaboration; 
; DNA = deoxyribonucleic acid; ECG = electrocardiogram; 
eGFR = estimated glomerular filtration rate; EOS = end of study; EOT = end of treatment; 
; FGF21 = fibroblast growth factor 21; 
FSH = follicle-stimulating hormone; GGT = gamma-glutamyl transferase; 
HBsAg = Hepatitis B surface antigen; HCV = Hepatitis C 
Virus; HDL-C = high-density lipoprotein-cholesterol; HHP = home health provider; HIV = Human Immunodeficiency Virus; hsCRP = high-sensitivity 
C-reactive protein; 
 INR = international normalized ratio; LDL-C = low-density lipoprotein-cholesterol; 
 
; non-HDL-C = non-high-density lipoprotein-cholesterol; MCV = mean corpuscular volume; MCH = mean corpuscular hemoglobin; MCHC = mean 
corpuscular hemoglobin concentration; MRI = Magnetic resonance imaging; MRI-PDFF = Magnetic Resonance Imaging – Whole Liver Proton Density Fat 
Fraction; 
 PK = pharmacokinetics; PT = prothrombin time; RNA = ribonucleic acid; RR = respiratory rate; 
RLP-C = remnant lipoprotein-cholesterol; SAE = serious adverse event; SoA = Schedule of Assessments; T3 = triiodothyronine; T4 = thyroxine; 
 
; TG = triglyceride; TSH = thyroid-stimulating hormone; V = Visit; VLDL-C = very low-density lipoprotein cholesterol; VLDL-TG = very 
low-density lipoprotein triglycerides; WBC = white blood cell; WOCBP = women of childbearing potential. 
Body posture during TG, BP, heart rate, RR and body temperature measurement needs to be the same (e.g., sitting, semi-erect, or supine) throughout the study 
with resting for at least 5 min prior to measurement at each study visit. 
Fasting blood samples should be taken after at least 12 hours of fasting. (It is recommended that subjects fast no more than 14 hours.) Fasting includes food and 
all beverages except for non-mineral water.  
Hematology: Hemoglobin, hematocrit, RBC count, reticulocyte count, MCV, MCH, MCHC, platelet count, WBC count, total neutrophils, eosinophils, 
monocytes, basophils, lymphocytes. 
Blood chemistry: BUN, creatinine (eGFR using CKD-EPI equation), fasting plasma glucose, calcium, sodium, potassium, chloride, total CO2 (bicarbonate), 
AST (SGOT), ALT (SGPT), ALP, GGT, total bilirubin, direct (conjugated) bilirubin, indirect (unconjugated) bilirubin, CK, uric acid, albumin, total protein.  
Coagulation panel: INR, aPTT, PT 
Urinalysis: pH, glucose, protein, blood, ketones, nitrites, leukocyte esterase, urobilinogen, urine bilirubin, microscopy 
a = If 
 is required, only assessment at that visit is TG measurement. 
b =
 
c = Symptom-driven physical assessment for applicable reported AEs. Can be conducted on site during a scheduled or unscheduled visit or by a home health 
provider (HHP) if identified on the delegation log. 
d = Height at screening 
 only  
e = Starting from randomization, pre-dose BP and pulse will be measured in duplicate. The first measurement will be initiated before dosing with 
sitting/semi-erect or supine position and resting for at least 5 minutes prior to measurements. BP and pulse are to be repeated at least 1 minute apart. Additional 
vital signs measurements may be done if clinically indicated. 
f = Coagulation panel will be done at 
 only. 
g = History of drug and/or alcohol abuse taken at screening 
 only. Urine drug test will be performed at 
 only. 
h= Alcohol breathalyzers will be used. Results >0.01% will be considered as positive. 
i = 
 
j = 
 
 
 
 
 
 
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iii. 
 
 
k = Subjects will be included in either the Intensive PK scheme (optional) or Trough PK scheme (all subjects). Table 5 details sampling times for the Intensive 
PK scheme. 
 For the Trough PK scheme, blood samples 
will be taken 
 
 Note: Where the Intensive PK schedule overlaps with the Trough PK schedule, only 1 PK measurement should be taken at those time points. 
l = Serology tests will include HBsAg, HCV, and HIV 1 and 2 antibodies. HBV and HCV determined by antibodies first and, if positive, by DNA/RNA. 
m = Ambulatory 24-hour urine collection for analysis of cortisol and creatinine to be initiated the day prior to 
. Subject will start collection 
24 hours before coming into the clinic and bring the sample to the visit. The 24-hour urine collection jug is dispensed at 
 so collection can be 
completed and brought to 
, respectively. The jug may be provided at the site or by an HHP. 
n = Archived for possible analysis of additional 
. This is optional and by subject consent.  
o =
 
 
p = Subjects randomized to BIO89-100 36 mg Q2W group will receive BIO89-100 at 
 
 
q = When IP is scheduled to be administered by an HHP, a subject’s spontaneous reports of AE/SAE will be reported to the site staff who will follow-up and 
evaluate the need for an unscheduled visit for AE assessment. The sites/HHP may take non-personally identifying photographs of potential injection site 
reactions (optional).  
r = Study coordinators should reinforce maintenance of eating and exercise habits without weight loss or gain of 5% for the duration of the study, and alcohol 
consumption restrictions 48 hours prior to fasting blood work.  
s = After a consenting subject’s 
, the site should preemptively schedule the baseline MRI-PDFF to occur within the following projected visit windows to 
ensure an MRI appointment is available to be performed after the 
 TG result ≥350 mg/dL is known. For the main cohort, MRI-PDFF is optional and should 
occur prior to 
. For the fibrate expansion cohort, MRI-PDFF of ≥ 6.0% is required for enrollment and should be performed between 
 
. The follow-up MRI at 
 should only be performed if the baseline liver MRI-PDFF value is ≥ 6.0%. MRI-PDFF is mandatory in the 
fibrate expansion cohort. 
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2. 
INTRODUCTION 
2.1. 
Study Rationale 
BIO89-100 is a glycoPEGylated analog of FGF21, 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 
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 (Li, et al. 2015; Ogawa, et al. 2007). Administration of exogenous FGF21 is 
being explored as a method to treat hypertriglyceridemia, which is associated with increased risk 
of AP and CVD. Severe hypertriglyceridemia, defined as fasting TG levels of greater than or equal 
to 500 mg/dL, is associated with an increased risk of AP, accounting for up to 10% of all AP 
episodes (de Pretis, 2018), and an increased risk of NAFLD, NASH, and CVD (Grundy, et al. 
2019). Clinical practice guidelines from both the American College of Cardiology/American Heart 
Association and from the American Association of Clinical Endocrinologists/American College of 
Endocrinology emphasize the importance of reducing TG in patients with levels above 
500 mg/dL. Current treatments for hypertriglyceridemia include fibrates, fish oils, niacin, and 
statins. While currently approved therapies are proven to reduce TG levels, the treatment goal of 
alleviating SHTG, i.e., reducing TG levels to below 500 mg/dL remains a significant challenge. In 
the Phase 3 studies with Vascepa® and Epanova®, ~50% of subjects did not have TG level 
reductions below 500 mg/dL at the end of the 12-week studies (Chowdhury, 2012, Chowdhury 
2014). Patients with TG levels above 500 mg/dL remain at risk for AP as well as cardiovascular 
and fatty liver diseases. Several non-clinical and clinical studies have shown that administration of 
various FGF21 analogs had beneficial effects on serum lipid and insulin-resistance as well as on 
liver fat (Zhang, et al. 2014, Gaich, et al. 2013, Sanyal, et al. 2018). A PD effect of BIO89-100 on 
TGs has been demonstrated, with significant reductions in TG levels in a non-human primate 
disease model and in the Phase 1a clinical study, TV47948-SAD-10122. 
 
 
 
 
 
 
 In Part 1 of the Phase 1b/2a 
study BIO89-100-002, subjects with NASH or NAFLD and high risk for NASH 
(hypertriglyceridemia was not an enrollment criterion) who received BIO89-100 at 27 mg QW 
showed a significant beneficial effect on the lipid profile, evaluated as percent change at Week 13 
(-28% decrease in TG, -16% reduction in non-HDL cholesterol and LDL cholesterol, p<0.05 for 
all comparisons). 
The current data suggest that BIO89-100 achieves potent and rapid reductions in TG levels 
through pathways mediated by FGF21, offering novel mechanisms of action. SHTG is associated 
with serious outcomes including risk of significant morbidity and mortality, and BIO89-100 has 
the potential for development as an important new therapy. 
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This Phase 2 study is designed to assess the efficacy, safety, and tolerability of QW or Q2W, SC 
dosing of BIO89-100 compared to placebo in subjects with SHTG. All subjects may be on a 
background therapy of prescribed fish oils or statins, but not on PCSK9 inhibitors, niacin, or any 
supplements, including non-prescribed fish oils, used to alter lipid metabolism. Subjects who are 
not taking prescription therapy to lower TG may also be enrolled. Subjects in the main cohort will 
not be on concurrent fibrate therapy. In contrast, subjects in the fibrate expansion cohort will 
remain on their current stable dose of fibrate therapy.  
2.2. 
Background 
2.2.1. 
Severe Hypertriglyceridemia 
Hypertriglyceridemia refers to elevated TG concentrations in the blood, usually measured after 
fasting but occasionally non-fasting. High TGs are defined as TG = 200 to 499 mg/dL and very 
high triglycerides, or SHTG as TG ≥500 mg/dL (Berglund, et al. 2012; Jellinger, et al. 2017). 
Subjects with elevated levels of serum TGs (≥500 mg/dL) are at increased risk of AP, NASH, 
NAFLD and CVD (Grundy, et al. 2019). Persistent hypertriglyceridemia is associated with the 
development of AP and CVD. For example, people with high versus low, non-fasting TG 
(580 mg/dL vs 70 mg/dL) were at 5.1-fold higher risk for myocardial infarction, 3.2-fold higher 
risk of ischemic heart disease, 3.2-fold higher risk for ischemic stroke, and 2.2-fold higher risk for 
all-cause mortality (Nordestgaard, 2016). Patients with SHTG are at increased risk of AP with a 
reported 5% and 10% to 20% lifetime risk of AP in patients with TG >1000 and >2000 mg/dL 
respectively (Blom, et al. 2018, Scherer, et al. 2014). Kiss et al., reported that elevated TG 
worsens the severity of AP and increases the risk for local and systemic complications, however, 
they reported no specific correlation of AP severity with extent of hypertriglyceridemia (Kiss, et 
al. 2018). The mechanisms by which SHTG causes pancreatitis are not fully understood but may 
relate to impairment of capillary perfusion in the pancreas, pancreatic cell necrosis and subsequent 
release and activation of pancreatic enzymes (Blom, et al. 2018). If and when pancreatitis will 
occur in an individual patient is unpredictable. Episodes of acutely-elevated TG levels, associated 
for instance with alcohol use or meals with excessive fat, could increase risk in patients with 
chronically high TG levels. In most patients who develop AP secondary to SHTG, the condition is 
mild and self-limiting, but ~20% of patients suffer severe attacks associated with prolonged 
hospitalization and significant morbidity and mortality. 
Given the high risk of AP associated with SHTG, early diagnosis is essential so that treatment can 
be initiated, including simple carbohydrate- and fat-restricted diet, use of medium-chain TG-rich 
foods and alcohol abstinence (Berglund, et al. 2012; Jacobson, et al. 2015). 
Current approved therapies for SHTG, as an adjunct to diet, include multiple fibrates (approved by 
the FDA from 1993 to 2009), niacin (extended release approved by the FDA in 1997), and omega-
3 fatty acids (approved in 2004, 2012, and 2014). Fibrates, niacin, and prescription omega-3 fatty 
acids, alone or in combination, can reduce TG by 20% to 50% in some patients with moderate to 
severe hypertriglyceridemia. However, minimal effects are seen in other patients, especially those 
with mutations in genes in the lipoprotein lipase pathway (Brisson, et al 2010). Statins have a 
modest TG lowering effect and are used in patients with moderately elevated TG levels, but they 
should not be used alone in patients with severe or very severe hypertriglyceridemia. 
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In patients at high risk for pancreatitis, fibrates remain first line drug treatment, but LDL-C levels 
may increase during therapy with fibrates which is a detrimental effect in this patient population. 
Fibrates are associated with an increase in the incidence of cholesterol gallstones, increases in 
transaminases and creatinine, among other safety and tolerability concerns (Lopid US Package 
Insert, 2017; Tricor US Package Insert, 2018). Further, due to increased risk of myopathy, the 
combination of gemfibrozil and statins should be avoided (Berglund, et al. 2014). 
The approvals of the omega-3-acids (namely Vascepa® and Epanova®) represent advances in the 
therapeutic options with improved risk:benefit profiles as compared to the fibrates and niacin 
formulations. While currently approved therapies are proven to reduce TG levels, the treatment 
goal of alleviating SHTG, i.e., reducing TG levels to below 500 mg/dL remains a significant 
challenge. In the Phase 3 studies with Vascepa® and Epanova®, ~50% of the subjects did not 
have TG level reduction below 500 mg/dL at the end of the 12-week studies (Chowdhury, 2012; 
Chowdhury, 2014). Patients with TG levels above 500 mg/dL remain at risk for AP as well as 
cardiovascular and fatty liver diseases. There remains a critical need for new therapies that will 
reliably reduce TG levels below 500 mg/dL rapidly after diagnosis. 
2.2.2. 
FGF21 
FGF21 is a peptide hormone, which is secreted mainly by the liver. The activation of 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 (Li, et al. 2015; Ogawa, et al. 2007). 
In patients with hypertriglyceridemia, circulating and/or tissue levels of FGF21 are increased, 
indicating the presence of FGF21 resistance, which can be overcome by administration of 
pharmacological doses of FGF21 (Galman, et al. 2008). On this basis, administration of 
exogenous FGF21 has been explored as a method to treat obesity-associated insulin-resistance 
disorders, including hypertriglyceridemia (Gaich, et al. 2013). 
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, which is likely a result of greatly 
increased in vivo half-life (Mu, et al. 2012). In 2 studies in humans, the effects of pegbelfermin 
(BMS-986036; a PEGylated analog of FGF21, which differs from BIO89-100 in PEG size, 
PEGylation site and technology, and location of mutations) were assessed separately in NASH or 
obese subjects. NASH subjects treated for 16-weeks with pegbelfermin experienced a statistically 
significant reduction in serum TGs versus placebo of 4.8% and 4.7% with doses of 10 mg/d and 
20 mg/week, respectively (Sanyal, et al. 2018). Obese subjects treated for 12 weeks with 
pegbelfermin experienced a ~20% reduction of serum TGs(p=0.037) at a dose of 20 mg/day 
(Charles, et al. 2019). 
Improvements in LDL-C, and HDL-C were also noted in the NASH study primarily with the 
10 mg daily dose. Finally, safety was considered favorable, with no deaths, treatment-related 
serious adverse events (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 severe in intensity. 
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2.2.3. 
BIO89-100 
BIO89-100 is a glycoPEGylated analog of FGF21 that has an N-terminal methionine residue, 
2-point mutations, and a single 20 kDa linear PEG covalently attached via a glycosyl moiety. Both 
point mutations are less likely to be exposed due to steric hindrance by the PEG domain, and 
thereby less likely to trigger an immune response. It has been evaluated in non-clinical 
pharmacology, PK, and toxicology studies and in a recently-completed, first-in-human, single 
ascending dose (SAD) study (TV47948-SAD-10122). 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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 SAD, has recently been completed. 
 
No significant safety issues have been identified. The most 
common reported AEs (≥2 in the pooled BIO89-100 group) were injection site reactions (all 
common terminology criteria for adverse events [CTCAE] Grade 1), headache, contact dermatitis, 
and nausea. There were no deaths, SAEs, or discontinuations, and review of ECG, clinical 
laboratories, and vital sign data did not reveal any concerning trends. There was a single CTCAE 
Grade 3 TEAE that was considered not treatment-related (elevated creatine phosphokinase level at 
EOS in a physical laborer). No other CTCAE Grade 3 or above TEAE was reported. 
Part 1 of 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 
 of 
BIO89-100 administered SC in subjects with NASH or with NAFLD who are at a high risk of 
NASH. Subjects were treated at QW SC doses of 3 mg, 9 mg, 18 mg, and 27 mg, and Q2W SC 
doses of 18 mg and 36 mg of BIO89-100 or placebo. Data from the MAD study Part 1 
demonstrated favorable safety and tolerability with repeated dosing of BIO89-100 for 12 weeks 
with evidence of clinical efficacy in NASH (clinically meaningful reductions in liver fat assessed 
by MRI-PDFF and in ALT). 
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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 
 
 
2.3.1. 
Risk Assessment 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
BIO89-100 is administered subcutaneously; ISRs are well-recognized with SC administration of 
biological agents. As with any biologic drug, exposure to BIO89-100 may potentially provoke an 
immune response with formation of 
 that may lead to loss of efficacy, neutralization of 
endogenous FGF21, or adverse effects (e.g., hypersensitivity reaction). 
 
 
 
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Increased appetite, which has been reported with other FGF21 analogs, occurred in 15.9% of 
BIO89-100-treated subjects and no placebo-treated subjects in Study BIO89-100-002 (Part 1) 
without a clear dose response. 
In a clinical study of another FGF21 analog, increased blood pressure and heart rate were reported. 
No clinically significant changes in vital signs or ECGs were observed in study 
TV47948-SAD-10122 or study BIO89-100-002 (Part 1). 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
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• 
 
 
 
 
 
 
 
 
 
 
 
 
 
2.3.2. 
Benefit Assessment 
Patients with SHTG are at increased risk of AP and cardiovascular comorbidities and require 
significant reductions in TG to lower the risk of AP. As a substantial proportion of patients have 
persistent hypertriglyceridemia despite the use of TG lowering medications, there remains a 
critical need for new therapies that will reliably reduce TG levels below 500 mg/dL rapidly after 
diagnosis. 
Long-acting FGF21 analogs are a promising mode of action for the treatment of SHTG; multiple 
FGF21 analogs have been shown to reduce levels of TG and other lipids in studies of subjects with 
NASH or obesity and Type 2 diabetes mellitus (Charles ED, 2019, Gaich G, 2013, Sanyal A, 
2019, Talukdar S, 2016). A PD effect of BIO89-100 on TGs has also been demonstrated with 
significant reductions in TG levels in the Phase 1a clinical study, TV47948-SAD-10122, and in a 
non-human primate disease model. Furthermore, BIO89-100 also offers the potential for broader 
metabolic benefits, including amelioration of fatty liver disease, reduction in LDL-C, increase in 
HDL-C, and improvements in glycemic control that may confer an additional advantage in 
patients with SHTG , many of whom have metabolic comorbidities. 
Taken together, treatment with BIO89-100 may have a positive impact on serum TG 
concentrations and other metabolic parameters in treated subjects.  
2.3.3. 
Overall Benefit-Risk Assessment 
Considering the potential benefits, potential risks, and risk mitigation measures that have been 
implemented, the benefit-risk profile of administering BIO89-100 to subjects with SHTG in study 
BIO89-100-221 is considered to be favorable. The overall benefit-risk associated with 
administration of BIO89-100 will be continually re-assessed with the emergence of additional 
data. 
Safety of trial participants is of primary importance. Subject participation in the trial during the 
COVID-19 pandemic requires that site personnel and study subjects follow key protective 
measures against COVID-19 infection, aligned with local requirements and recommendations. 
89bio and participating study sites will continually monitor the conditions of the pandemic as they 
may affect the subjects’ safe participation in the trial. 89bio and participating study sites may 
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implement changes to the protocol based on an overall risk assessment that includes 
considerations related to COVID-19 that may result in changes or alternatives regarding in-clinic 
visits, methods of performing assessments, monitoring, data collection, and/or trial participation. 
Provisions concerning IP administration related to potential impacts due to COVID-19 are 
described in Section 7.1.  
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3. 
OBJECTIVES AND ENDPOINTS  
The objectives and endpoints are the same for the main cohort and fibrate expansion cohort. 
Objectives 
Endpoints 
Primary 
 
• To determine the effect of 
BIO89-100 on serum TG 
levels in subjects with SHTG 
(TG ≥500 mg/dL)  
• Percentage change in serum TG from baseline to 
Week 8  
Secondary 
 
• To determine the effect of 
BIO89-100 on selected serum 
lipids and lipoproteins  
• Achieve TG <500 mg/dL at Week 8 
• Percentage change in VLDL-C, LDL-C, 
non-HDL-C, HDL-C, VLDL-TG, ApoB, RLP-C 
from baseline to Week 8 
• To determine the effect of 
BIO89-100 on hsCRP 
• Percentage change in hsCRP from baseline to 
Week 8 
• To determine the effect of 
BIO89-100 on metabolic 
markers 
• Percent change in fasting plasma glucose, 
adiponectin, and body weight from baseline to 
Week 8 
• To characterize BIO89-100 
PK 
• Serum BIO89-100 concentration  
• PK parameters in Intensive PK subgroup  
o Cmax within a dosing interval 
o AUC0-tau within a dosing interval 
o tmax 
o t½ 
• Additional PK parameters may be calculated if 
also deemed appropriate. 
• To characterize BIO89-100 
PD profile as assessed by 
MRI-PDFF 
• Percentage change and change from baseline in 
hepatic steatosis using MRI-PDFF  
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Objectives 
Endpoints 
Exploratory 
 
• 
 
 
  
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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Objectives 
Endpoints 
Safety Endpoints 
 
• To evaluate the safety and 
tolerability of BIO89-100 
• Frequency of TEAEs 
• Frequency of TESAEs 
• Number of subjects who discontinue due to 
TEAEs 
• 
 
• To determine the effect of 
BIO89-100 on liver function 
biomarkers 
• Change in ALT and AST from baseline to Week 8 
• Other safety assessments 
• Incidence and shifts of clinically significant vital 
signs, physical examination findings, ECG data 
and laboratory abnormalities; safety laboratory 
evaluations include CBC, blood biochemistry, 
24-hour urine cortisol, and urinalysis. 
• 
 
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4. 
STUDY DESIGN 
4.1. 
Overall Design 
Study BIO89-100-221 is a randomized, double-blind, placebo-controlled, Phase 2 study to 
evaluate the efficacy, safety, tolerability, PK and PD profiles, and 
 of BIO89-100 
administered SC for 8 weeks. The main cohort will include ~90 subjects with SHTG without 
concurrent fibrate therapy and will consist of 5 treatment groups to compare 4 dose 
levels/regimens of BIO89-100 versus placebo (Figure 1). The fibrate expansion cohort will 
include ~36 subjects with SHTG on stable background fibrate therapy and with an MRI-PDFF 
≥6.0% and will consist of 2 treatment groups comparing one dose regimen of BIO89-100 versus 
placebo (Figure 2). Investigators whose sites are obtaining MRIs should assess if subjects on 
background fibrate therapy are appropriate for the main cohort (acceptable to washout fibrate 
therapy) or for the fibrate expansion cohort. For participation in the fibrate expansion cohort, 
investigators should first discuss the potential subject with the Medical Monitor 
The study includes lifestyle stabilization, TG qualification, treatment, and follow-up periods. 
After signing the informed consent form and confirmation of general eligibility, subjects will 
undergo further screening assessments to confirm eligibility over a period of ~6 to 9 weeks. 
Generally, eligible subjects will enter a 4- or 6-week lifestyle stabilization period (maximum of 
6 weeks for subjects stopping lipid-altering therapy, minimum of 4 weeks for subjects already on 
stable, protocol-accepted lipid-altering therapy), followed by a ~2-week TG qualification period 
with qualifying TG measurements
 
 
Individuals who do not meet the criteria for participation in this study (screen failures other than 
for TGs) may be re-screened, 
, if the reason for screen failure is deemed 
temporary and is resolved prior to re-screening (e.g., respiratory infection). Re-screened subjects 
will sign a new consent and be assigned a new ID. Subjects can only re-screen once. Re-screening 
subjects who have maintained the lifestyle stabilization period may be allowed to forgo repeated 
tests 
 at the discretion of the Medical Monitor. Re-screening after COVID-19 
infection requires Medical Monitor approval.  
Subjects fulfilling all of the qualifying criteria and none of the exclusionary criteria will be 
randomized 1:1:1:1:1 for the main cohort 
 and enter a double-blind, 8-week treatment 
period during which they will be dosed with 1 of 4 doses of BIO89-100 (9 mg QW, 18 mg QW, 
27 mg QW, or 36 mg Q2W) or placebo.  
Enrollment will begin in the fibrate expansion cohort, with randomization 1:1 
 to a 
double-blind, 8-week treatment period during which subjects will be dosed with BIO89-100 
27 mg or placebo QW.  
Randomization in the main cohort will be stratified by TG <750 mg/dL or ≥750 mg/dL 
(8.47 mmol/L) and by whether or not the subject is receiving background therapy (e.g., 
prescription fish oil and/or statins). Randomization in the fibrate expansion cohort will be 
stratified by TG <750 mg/dL or ≥750 mg/dL (8.47 mmol/L) only. 
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Treatment with IP will be given at the Investigator site, QW, SC into the abdomen. To maintain 
the blind, all subjects will receive 2 SC injections 
. Subjects 
randomized to 36 mg Q2W will receive placebo at visits between BIO89-100 Q2W 
administrations. Intensive PK is planned to be assessed for a subset of subjects (optional 
participation with no more than 45 subjects). Treatment assignment and results of the post-
baseline lipid panels (TG, 
, LDL-C, HDL-C, non-HDL-C, VLDL-C, VLDL-TG, 
, 
ApoB, 
, RPL-C, 
), post-baseline MRI-PDFF, 
, and PK data will be blinded to 
subjects, sites, study team, and 89bio.  
The efficacy, safety, and tolerability of the treatments will be monitored throughout the study as 
described in the objectives (Section 3).  
Subjects who terminate treatment early will be requested to have an EOT visit 
 
 and an EOS visit 4 weeks later. All subjects who complete the treatment 
period will enter a 4-week follow-up period. 
4.2. 
Scientific Rationale for Study Design 
BIO89-100-221 a Phase 2, randomized, double-blind, placebo-controlled study designed to assess 
the efficacy, safety and tolerability of different doses and dose regimens (QW or Q2W) of SC 
BIO89-100 compared to placebo in subjects with SHTG. The study will separately evaluate the 
response to BIO89-100 in subjects who are not on concurrent fibrate therapy and subjects who 
remain on their current fibrate therapy. The primary and secondary efficacy endpoints, safety, and 
PK are characteristic for this type of study. Results from this study will provide proof-of-concept 
data to inform dose selection in the future Phase 3 clinical program. 
Fibrate therapy is commonly used to lower TGs in patients with SHTG. Fibrates act by agonizing 
PPAR⍺, which stimulates the release of FGF21 in the liver. It is unknown if the levels achieved by 
exogenous FGF21 supplementation with BIO89-100 will be greater than, or have an add-on effect 
to, those achieved by fibrate-induced PPAR⍺ agonism alone.  
One potential additional benefit of BIO89-100 is reduction in liver fat content. Fatty liver has been 
shown to be significantly associated with moderately severe or severe acute pancreatitis (Yoon, et 
al. 2017). In Study BIO89-100-002 (Part 1), BIO89-100 (27 mg QW) reduced liver fat as assessed 
by MRI-PDFF up to 60%. Fibrates do not reduce liver fat content. To evaluate this additional 
potential benefit, only subjects with liver fat content ≥6.0% by MRI-PDFF at baseline will be 
enrolled into the fibrate expansion cohort.  
4.3. 
Justification for Dose 
Five groups of SHTG subjects are planned to be dosed, SC × 2 per visit, into the abdomen, with 
either placebo or 1 of 4 doses of BIO89-100 (9 mg QW, 18 mg QW, 27 mg QW, or 36 mg Q2W) 
(Figure 1). 
These dose regimens were selected based on non-clinical pharmacological and PK models mainly 
in spontaneously diabetic monkeys, review of clinical PK, and efficacy data from the single 
ascending dose clinical study TV47948-SAD-10122 and PK modeling. 
 
 
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The selected dose of 27 mg QW was chosen for the fibrate expansion cohort based on the results 
observed in subjects with NASH or NAFLD and high risk for NASH in Study BIO89-100-002 
(Part 1), in which 10 subjects were randomized to that dose.  
 
 
 
 
  
The benefit/risk assessment for BIO89-100 at the chosen doses is presented in Section 2.3. Based 
on these data, the dose regimens are anticipated to be safe, and 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 
assessed in subsequent studies. 
4.4. 
End of Study Definition 
A subject is considered to have completed the study if he/she has completed all phases of the study 
including the last scheduled procedure shown in the Schedule of Assessments (SoA) (Table 2). 
The EOS is defined as the date of the last visit of the last subject in the study. 
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5. 
STUDY POPULATION 
5.1. 
Inclusion Criteria 
Subjects are eligible to be included in the study only if all of the following criteria apply: 
1. Subject understands the study design and has been informed of the investigational nature 
of the study. Subject has given voluntary, written, informed consent to take part in this 
study. 
2. Subject agrees to be compliant and is capable of completing the required study 
assessments. 
3. Male or female age ≥21 to ≤75 years at time of signing informed consent. 
4. All subjects (male or female) who are of childbearing potential must agree to use 
highly-effective, double contraception (with male/female partners) during the study. 
Double contraception is defined as use of a condom by the male partner, combined with 
use of 1 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® 
e. Documented evidence of tubal ligation at least 6 months prior to the screening visit).  
Male subjects with a history of vasectomy should also use condoms as double 
contraception during the study. Use of highly-effective, double contraception must 
continue for 30 days or 5 half-lives (whichever is longer) after the last dose of IP. Female 
subjects must 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 IP is acceptable if it is the subject’s regular practice. 
5. Females of childbearing potential must have a negative serum pregnancy test at screening 
 and agree to undergo a urine pregnancy test at 
, and a serum 
pregnancy test at end of the study 
. 
6. 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 (e.g., total hysterectomy, 
partial hysterectomy, or oophorectomy). 
7. Serum TG criteria meeting all of the following criteria: 
a. A historical documented TG ≥400 mg/dL (4.52 mmol/L) in the past 5 years from 
screening 
 or currently on lipid-modifying therapy due to SHTG at screening 
. 
b. Mean of 2 screening fasting serum TGs ≥500 mg/dL (5.65 mmol/L) and ≤2000 mg/dL 
(22.60 mmol/L). The target for the study is 500 mg/dL. Due to TG variability, up to 
10% of subjects enrolled may have a mean qualifying TG level between 475 and 
500 mg/dL. Mean screening fasting serum TG is defined as the mean of TG 
 
 if the average of TG at 
 is <500 mg/dL, or 
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>2000 mg/dL. The repeat measure of TG is permitted at a minimum of 7 days apart at 
.  
c. If 1 of the 2 qualifying TGs is <400 mg/dL, the difference between 2 TG should be 
<300 mg/dL. Note: If a subject has an individual fasting TG value of <350 mg/dL at 
any one visit or mean of two fasting TG levels <475 mg/dL, they will be considered a 
screen fail and not eligible to be re-screened. 
8. Willing to maintain current eating and exercise habits from time of signing the informed 
consent and for the duration of the study. (See also Inclusion Criterion #11 and Exclusion 
Criterion #10 regarding alcohol consumption and Exclusion Criterion #4 regarding weight 
changes.)  
9. Concomitant medication may include prescription fish oil (including purified 
eicosapentaenoic acid [EPA] with or without docosahexaenoic acid [DHA]) and/or statin 
with or without ezetimibe (none of which will be supplied as part of study) as long as the 
subject has been clinically stable (per Principal Investigator [PI] judgment) while on stable 
dose for 
. 
10. Medicines known to exacerbate hypertriglyceridemia (such as certain beta blockers, 
thiazides, estrogens, bile acid sequestrants) should be stable 
 
) and remain stable during the study. 
11. Subject is willing to refrain from alcohol consumption for ≥48 hours before each study 
visit. 
12. MRI-PDFF of ≥ 6.0% for subjects screened for the fibrate expansion cohort. Note: Those 
who cannot undergo MRI for any reason (e.g., claustrophobia, metal implants) cannot 
participate in the fibrate expansion cohort. 
5.2. 
Exclusion Criteria  
Subjects are excluded from the study if any of the following criteria apply: 
1. Females who are 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 IP.  
2. Uncontrolled or newly diagnosed (< 3 months since diagnosis at time of Screening) 
hypertension. Subjects with well controlled hypertension who are clinically stable may 
enroll if they have been on a stable dose of antihypertensive medications for at least 
2 months before Screening. Subjects who are taking >2 antihypertensive medications may 
be eligible with Medical Monitor (or designee) approval. 
3. Body mass index (BMI) >45 kg/m2 at screening 
. 
4. Weight change ≥5% in 3 months prior to screening 
 or weight change ≥5% between 
screening 
. 
5. Central laboratory hemoglobin levels below the lower limit of normal (<12.0 g/dL for 
males, <11.0 g/dL for females), or dialysis, lipid apheresis, plasma exchange, blood 
transfusion, or blood donation/blood loss of ≥400 mL at least a month prior to and during 
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the screening period or anticipation of these procedures during the period of the study and 
up to 30 days afterwards. 
6. A history of symptomatic gallstone disease unless treated with cholecystectomy and/or 
pancreatitis within 5 years from screening will be excluded from the study. 
7. ALT or AST >3× upper limit of normal (ULN) at screening 
. 
8. Total bilirubin exceeds ULN at screening 
, unless prior diagnosis and documentation of 
Gilbert’s syndrome in which case total bilirubin must be ≤3 mg/dL. 
9. Creatine kinase (CK) > 5 × ULN at screening 
. 
10. 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 screening 
 or had positive alcohol test at screening 
. A unit of alcohol is defined as 14 
gram or as 355 mL of beer (5% alcohol by volume [ABV]), 1 glass of wine (150 mL; 12% 
ABV), or 1 shot of hard liquor (45 mL; 40% ABV). During the study, subjects will receive 
counseling and will be encouraged to abstain from alcohol. 
Alcohol intake will be limited to 2 units of alcohol per day for men and 1 unit of alcohol 
per day for women. Subjects must be willing to abstain from alcohol use 48 hours prior to 
study visits. 
11. Concomitant use of PCSK9 inhibitors, niacin, or any supplements, including 
non-prescription, non-pharmaceutical strength fish oils, used to alter lipid metabolism. 
Previous treatment with a PCSK9 inhibitor will need to have ceased >12 weeks before 
screening with no intention to reinitiate treatment for the duration of the trial. Other lipid-
modifying supplements including, but not limited to, red rice yeast supplements, garlic 
supplements, soy isoflavone supplements, sterol/stanol products, or policosanols are not 
permitted.  
a. Subjects participating in the main cohort previously treated with a fibrate will need to 
agree to cease fibrate treatment for the duration of this trial.  
b. Subjects enrolled in the fibrate expansion cohort will need to maintain their current 
stable dose of fibrate therapy (≥4 weeks before screening 
). 
12. Previous long-term (>4 weeks) use of systemic steroid (glucocorticoid) medications such 
as prednisone within 12 months of screening 
. Inhaled or topical corticosteroids are 
permitted. 
13. Reduced renal function (eGFR ≤ 55 mL/min/1.73 m2 calculated using chronic kidney 
disease epidemiology collaboration [CKD-EPI] equation). 
14. Positive for hepatitis B virus (HBV), hepatitis C virus (HCV) or human immunodeficiency 
virus (HIV). HIV, HBV and HCV determined by antibodies first and, if positive, 
confirmed by DNA/ribonucleic acid (RNA). Subjects who have tested positive for 
COVID-19 who are asymptomatic or who have active COVID-19 infection are excluded. 
Note: subjects who had COVID-19 and recovered without sequelae or tested positive 
without symptoms at least 8 weeks prior to screening may be included.  
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15. History of substance use disorder, or any other substance dependence (with the exception 
of caffeine) as defined by the Diagnostic and Statistical Manual of Mental Disorders 4th 
edition Text Revision (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. 
16. Participation in a previous clinical trial or exposure to another investigational drug within 
12 weeks or 5 half-lives, whichever is longer, prior to screening.  
17. Previous exposure to an FGF21 analog 
 
r FGFR1 activating product or known sensitivity to PEG or any of the 
excipients. 
18. Type 1 diabetes mellitus (T1DM). 
19. Diagnosis of Type 2 diabetes mellitus (T2DM) <6 months prior to screening.  
20. Subjects with T2DM diagnosed ≥ 6 months prior to screening must have a HbA1c < 9.5% 
and if receiving antidiabetic medications, must be on a stable regimen before screening. A 
stable regimen is defined as no addition or discontinuation of antidiabetic medications, but 
dose adjustments or switching to another medication in the same class at the same relative 
dose per standard of care are allowed. Insulin, dipeptidyl peptidase IV (DPP-IV) 
antagonists and other oral medications are allowed if subject has been on a stable regimen 
for at least 3 months. Glucagon-line peptide 1 (GLP-1) agonists and sodium glucose 
co-transporter 2 (SGLT2) inhibitors are allowed if subject has been on stable doses for 
6 months. Use of thiazolidinediones such as pioglitazone, which can reduce serum TGs, 
are NOT allowed in this study. 
21. History of malignancy within 5 years prior to screening 
 other than successfully treated 
basal or squamous cell carcinoma or localized cervical carcinoma.  
22. Inadequately controlled thyroid disorders. Subjects on thyroid replacement should be on 
stable doses for at least 2 months prior to screening. 
23. Subjects with known lipoprotein lipase impairment or deficiency (Fredrickson Type 1), 
apolipoprotein C-II deficiency, or familial dysbetalipoproteinemia (Fredrickson Type 3). 
24. Clinically or otherwise documented cardiovascular or cerebrovascular disease including 
clinically significant anomalies of rhythm or pattern of ECG or New York Heart 
Association Class II to IV heart failure within 12 months of screening 
 that, in the 
judgment of the Investigator, could affect the safety of the subject or their ability to 
comply with the study requirements.  
25. History of bone trauma, fracture, or surgery within 2 months of screening or other bone 
disorders, such as osteoporosis, osteomalacia, and known untreated severe vitamin D 
deficiency (serum 25-hydroxy-vitamin D ≤5 ng/mL). 
26. Subject who cannot fast for study procedures for any reason per Investigator’s assessment 
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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27. An employee of the investigational center or has a family member who is involved with 
the conduct of this study. 
28. Any other condition(s) that would compromise the safety of the subject or compromise the 
quality of the clinical study, as judged by the Investigator. 
5.3. 
Lifestyle Considerations 
Beginning at the screening visit, all subjects will enter a diet, lifestyle, and medication 
stabilization period. Subjects should maintain their current eating and exercise habits as of the 
time they signed the ICF for the duration of the study. All subjects should be instructed to refrain 
from excessive alcohol consumption.  
5.3.1. 
Meals and Dietary Restrictions 
Study coordinators should reinforce maintenance of eating and exercise habits without weight loss 
or gain of 5% for the duration of the study, and alcohol consumption restrictions 48 hours prior to 
fasting blood work. All subjects will be required to fast for at least 12 hours (it is recommended 
that subjects fast no more than 14 hours) before visits requiring fasted blood sampling. Subjects 
will be instructed to attempt to remain consistent in how long they fast for the duration of the 
study. For the purposes of this study, fasting is defined as nothing by mouth except non-mineral 
water (and any essential medications). 
5.3.2. 
Alcohol 
Beginning at the screening visit, all subjects should be instructed to refrain from excessive alcohol 
consumption, limited to 2 units of alcohol per day for men and 1 unit of alcohol per day for 
women. Subjects will abstain from alcohol for 48 hours before each study visit. If the subject’s 
alcohol test is positive at a scheduled visit where lipid panels will be measured, the subject will be 
instructed to come back for an unscheduled visit for lipid panel measurement prior to or at the next 
scheduled visit. 
5.3.3. 
Activity 
Beginning at the screening visit, all subjects should not alter their normal activity routines for the 
duration of the study. 
5.4. 
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 failures other than 
for TGs) may be re-screened, at least 7 days later, if the reason for screen failure is deemed 
temporary and is resolved prior to re-screening (e.g., high blood pressure). Re-screened subjects 
will sign a new consent and be assigned a new ID. Subjects can only re-screen once. Re-screening 
subjects who have maintained the lifestyle stabilization period may be allowed to forgo repeated 
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tests 
 at the discretion of the Medical Monitor. Re-screening after COVID-19 
infection requires Medical Monitor approval.  
Retesting of laboratory tests is allowed when results are deemed spurious or incomplete. 
6.
INVESTIGATIONAL PRODUCT
Investigational product is defined as BIO89-100 or matching placebo, intended to be administered 
to a study subject according to the study protocol. 
6.1. 
Investigational Product(s) Administered 
Investigational product will be administered SC to the abdomen by qualified study personnel. 
Each administration will consist of 2 SC injections.  
Arm 
Active arm 
Placebo arm (control) 
Intervention 
BIO89-100 
Matching Placebo 
Type 
Biologic 
Chemical solution 
Dose 
Formulation 
 
 
  
 
 
 
Unit Dose 
Strength(s) 
Mg 
Not applicable 
Dosage Level(s)a 
9 mg, 18 mg, 27 mg QW 
36 mg Q2W 
NA; matching placebo will be injected 
at matching frequency per protocol. 
Route of 
Administration 
SC injection 
SC injection 
Sourcing 
Provided centrally by 89bio 
Provided centrally by 89bio 
Packaging and 
Labeling 
BIO89-100 is supplied as a sterile, 
preservative-free, frozen liquid 
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 liquid 
formulation in a single-use Type 1 clear 
glass vial for SC injection. Each vial 
will be labeled as required per country 
requirement. 
Abbreviations: QW = every week; Q2W = every 2 weeks. 
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. 
6.2. 
Administration Instructions 
On-site administration: The SC injection of the IP should be performed by a limited number of site 
staff specifically trained in the administration of SC injections. Injections will be administered in 
the abdominal area only. Details on injection of IP will be provided in the relevant study manual. 
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After administration of the IP, the subject should remain resting for ~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). 
Home administration: Home administration of IP may occur on 
. 
Home health visits conducted by qualified HHPs for IP dosing will follow the above 
administration protocol. Further details are provided in the relevant study manual.   
6.3. 
Preparation/Handling/Storage/Accountability 
BIO89-100 will be stored and prepared in appropriate conditions as indicated in the relevant study 
manual. 
• The Investigator or designee must confirm appropriate temperature conditions have 
been maintained during transit for all IP received and any discrepancies are reported 
and resolved before use of the IP. 
• Only subjects enrolled in the study may receive IP and only authorized study staff or 
HHPs may supply or administer IP. All IP on site 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 on-site IP accountability, reconciliation, and record maintenance (i.e., 
receipt, reconciliation, and final disposition records). 
• Further guidance and information for IP shipping, preparation, and administration and 
the final disposition of unused IPs are provided in the relevant manuals for on-site and 
home administration. 
6.4. 
Measures to Minimize Bias: Randomization and Blinding 
Eligible subjects will be randomized in the order they are enrolled into the study. Subjects in the 
main cohort will be randomized in a 1:1:1:1:1 ratio to 1 of 4 doses of BIO89-100 (9 mg QW, 
18 mg QW, 27 mg QW, or 36 mg Q2W) or placebo. Subjects in the fibrate expansion cohort will 
be randomized in a 1:1 ratio to BIO89-100 27 mg or placebo QW. 
To maintain the blind, each subject will receive 2 injections QW including subjects randomized to 
36 mg Q2W who will receive placebo injections at alternate Q2W visits. 
Randomization in the main cohort will be stratified by TG <750 mg/dL or ≥750 mg/dL 
(8.47 mmol/L) and by whether or not the subject is receiving background therapy (e.g., 
prescription fish oil and/or statins). Randomization in the fibrate expansion cohort will be 
stratified by TG <750 mg/dL or ≥750 mg/dL (8.47 mmol/L) only. All subjects will be centrally 
assigned to randomized IP using an Interactive Voice/Web Response System (IVRS/IWRS). 
Before the study is initiated, the log-in information and directions for the IVRS/IWRS will be 
provided to each site. 
The study will be conducted under double-blind conditions. The subjects, PI , other study 
personnel involved with subject assessments, and 89bio will remain blinded to the actual treatment 
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assignments of the subjects, post-baseline lipid panels (TG, 
, LDL-C, HDL-C, Non-HDL-C, 
VLDL-C, VLDL-TG, 
, ApoB, 
, RPL-C, 
), 
, and PK data. Blinded IP will 
be provided by a clinical supplies vendor and shipped to the study site. IP may be delivered to the 
subject’s home for home administration by an HHP. The SC administration of the blinded IP will 
be performed by trained study staff, and only blinded staff will be involved with subject 
assessments. 
6.5. 
Emergency Unblinding 
At the initiation of the study, Investigators will be instructed on the method for breaking the blind. 
The method will be an electronic process in the IWRS/RTSM system. Blinding codes should be 
broken only in exceptional circumstances when knowledge of the actual treatment code is 
essential for further management of the subject. Investigators are encouraged to discuss with the 
Medical Monitor if they believe that unblinding is necessary. When the blinding code is broken, 
the reason must be fully documented and subject will need to discontinue study drug and complete 
the final study procedures.  
6.6. 
Investigational Product Compliance 
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 IP and study subject 
identification will be confirmed by a member of the study site staff other than the person 
administering the IP. For doses administered by an HHP, administration details will be provided to 
the study staff for documentation. 
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 Medical 
Monitor if the dosing schedule changes markedly compared to the original schedule. 
6.7. 
Concomitant Therapy 
Any medication or vaccine (including over-the-counter or prescription medicines, vitamins, and/or 
herbal supplements) or other specific categories of interest that the subject has received within 
3 months from Screening 
 (prior therapy), is receiving at the time of enrollment, or receives 
during the study must be recorded along with: 
• Reason for use 
• Dates of administration including start and end dates 
• Dosage information including dose and frequency 
The Medical Monitor should be contacted if there are any questions regarding concomitant or 
prior therapy. 
Concomitant medication may include prescription fish oil (including purified EPA with or without 
DHA) and/or statin with or without ezetimibe (none of which will be supplied as part of study) as 
long as the subject has been clinically stable (per PI judgment) while on stable dose for ≥4 weeks 
before screening 
 and will maintain stable dose during the duration of the study. 
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Elevated levels of FGF21 have been linked to reduced cytochrome P450 3A (CYP3A) mRNA 
expression and activity in NAFLD patients (Woolsey et al, 2016). The underlying mechanism has 
not been fully elucidated and whether such effects will be observed in SHTG patients is unknown. 
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. 
COVID-19 vaccine, if available, will be allowed but subjects are prohibited from participation in 
interventional COVID-19 vaccine clinical trials.   
6.7.1. 
Prohibited Medications/Therapies 
Subjects must abstain from taking lipid-modifying supplements after the start of screening until 
completion of the EOS visit, unless, in the opinion of the Investigator and the Medical Monitor, 
the medication is required. 
Other prohibited therapies include: 
• Use of PCSK9 inhibitors, niacin, or any supplements, including non-prescription, non-
pharmaceutical strength fish oils, used to alter lipid metabolism. For subjects enrolled 
in the main cohort, use of fibrates is prohibited. 
• Medicines known to exacerbate hypertriglyceridemia (such as certain beta blockers, 
thiazides, estrogens, bile acid sequestrants) should be stable (≥4 weeks before 
screening 
 and remain stable during the study. 
• Other lipid-modifying supplements including, but not limited to, red rice yeast 
supplements, garlic supplements, soy isoflavone supplements, sterol/stanol products, or 
policosanols. 
• Long-term (>4 weeks) use of systemic steroid (glucocorticoid) medications such as 
prednisone. Inhaled and topical steroids are acceptable. 
• Use of thiazolidinediones such as pioglitazone  
• Any IP, FGF21 analog
 
or FGFR1 activating product. 
6.7.2. 
Blinded Alerts and Rescue Medications/Therapies 
Lipid panel results including TG are blinded during the study. If a subject has a measured fasting 
TG >2000 mg/dL, a blinded alert will be sent to the Investigator noting that the TG alert criteria 
has been met for that subject. The Investigator will contact the subject for the following: 
• Investigator to confirm and ensure the subject is managed and compliant with the 
allowed standard of care therapies for optimal hypertriglyceridemia management. 
• Investigator to confirm the subject was fasting for at least 12 hours and was absent 
from alcohol for 48 hours prior to the TG triggered blinded alert was measured. 
• Investigator to schedule the subject for an unscheduled fasting TG measurement. The 
subject should fast for at least 12 hours (and recommended no more than 14 hours) and 
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have no alcohol 48 hours prior to the planned unscheduled visit TG measurement. The 
subject should have the unscheduled fasting TG measurement prior to or at the next 
scheduled visit. 
If the subject has 2 consecutive blinded TG alerts, the Investigator should assess whether it is 
clinically safe for the subject to continue receiving IP. 
6.8. 
Stopping Rules 
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 
pandemic. 
6.8.1. 
Monitoring and Discontinuation for Suspected Drug-induced Liver Injury (DILI) 
Criteria 
A subject will be additionally monitored if they meet any of the following liver-related criteria. 
Baseline value is defined as the last ALT and AST values performed prior to or on the 
 
. 
New transaminase elevations in subjects with baseline value within the normal range:  
For new elevations in transaminases greater than 2× ULN, repeat measurement should be 
performed within 48 to 72 hours. 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.  
Drug should be discontinued, and the subject followed until resolution of symptoms or 
signs in the following situations: 
• ALT or AST >8× ULN 
• ALT or AST >5× ULN for more than 2 weeks 
• ALT or AST >3× ULN and (TB >2× ULN or INR >1.5) 
• ALT or AST >3× 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 greater than 2× baseline value or total bilirubin 
>1.5× ULN, repeat measurement should be performed within 48 to72 hours. 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. 
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Drug should be discontinued and the subject followed until resolution of symptoms or signs in the 
following situations described in Table 3: 
Table 3: 
Discontinuation Criteria in Subjects with Abnormal Baseline ALT or AST 
Values 
Baseline value of ALT/AST 
Criteria to discontinue investigational product 
<2× 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 >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 >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 = International Normalized ratio; 
ULN = Upper limit of normal 
Close monitoring for suspected DILI includes: 
• Repeating liver enzyme and serum bilirubin tests 2 or 3 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 non-prescription medications 
and herbal and dietary supplement preparations), alcohol use, recreational drug use, 
and special diets. 
• Ruling out acute viral hepatitis types A, B, C, D, and E, autoimmune or alcoholic 
hepatitis, hypoxic/ischemic hepatopathy; and biliary tract disease. 
• 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 subject lives in a remote area, laboratory testing can be performed locally, and the 
results should be promptly communicated to the Investigator site. 
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7.
INTERRUPTION OR DISCONTINUATION OF
INVESTIGATIONAL PRODUCT AND SUBJECT
DISCONTINUATION/WITHDRAWAL
7.1. 
COVID-19-Related Risk Mitigation and Interruption of
Investigational Product Administration
The following are provided as mitigations in the event of potential disruptions to study conduct 
secondary to COVID-19 infection or related control measures: 
•
Dose interruption resulting in missing 1 dose is acceptable at any time during the
dosing period.
•
Interruption in IP administration and other study procedures (collectively “study
interruption”) will be allowed for up to 2 weeks up to Week 5.
•
Dose interruption for 2 weeks after Week 5 is not allowed and will require
discontinuation of treatment as resuming the dose would impact the interpretation of
the primary endpoint.
•
Study interruptions longer than 2 weeks at any time during the dosing period will lead
to discontinuation of treatment and the subject’s participation in the study.
•
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
and will discuss the assessment with the Medical Monitor.
•
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.
•
If clinical laboratory tests are not part of the protocol-defined procedures for the first
dosing visit after study interruption, unscheduled hematology and clinical biochemistry
panels may be obtained prior to dosing as needed and at Investigator discretion and
sent to the central lab. Results from both local and central labs should be documented
in EDC.
•
If circumstances related to the COVID-19 pandemic or other extenuating
circumstances preclude a visit to the investigative site, alternative means for
completing study procedures and assessments may be considered at Investigator
discretion, in accordance with local guidance/regulations and with Medical Monitor
approval. For example, remote verification of source data may be instituted. Also,
safety assessments may be conducted via telemedicine or phone contact or laboratory
tests may be obtained at a certified local lab. Local analysis can be used for safety
decisions.
Efforts should be made to obtain EOT assessments in subjects who are not willing or able 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 the COVID-19 
pandemic. The EOT visit should be scheduled as soon as possible in these scenarios, whenever 
possible.  
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• 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 qualified subjects who have not yet received their first dose at a site and that 
can foresee a near-term disruption by the 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. Re-screening or 
retest of certain laboratory tests may be required prior to start of dosing, pending 
Medical Monitor evaluation.  
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 
eCRF 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). 
7.2. 
Discontinuation of Investigational Product 
In rare instances, it may be necessary for a subject to permanently discontinue IP. If IP is 
permanently discontinued, subjects will be requested to have an EOT visit within 7 days of 
terminating the treatment and an EOS visit 4 weeks later. The subject will remain in the study to 
be evaluated for safety tolerability, and PK/PD assessments. The reason for subject withdrawal 
from the IP will be recorded in the eCRF. See the SoA (Table 2) for data to be collected at the 
time of discontinuation of IP (EOT visit). 
7.3. 
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’s withdrawal of consent 
• Sponsor termination or suspension of the study 
• Lost to follow-up 
• A COVID-19-related dose interruption for 2 weeks or longer after 
 or 
study interruptions longer than 2 weeks at any time during the dosing period 
The reason for subject’s withdrawal from the study will be recorded in the eCRF.  
At the time of withdrawal from the study, subjects will be requested to have an EOT visit within 
7 days of terminating the treatment, and an EOS visit 4 weeks later, as shown in the SoA (Table 
2). 
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7.4. 
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 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 and/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. 
• Should the subject continue to be unreachable, he/she will be considered to have 
withdrawn from the study.  
Discontinuation of specific sites or of the study is handled as part of Appendix 1 (Section 10.1.8). 
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8. 
STUDY ASSESSMENTS AND PROCEDURES 
• Study procedures and their timing are summarized in the SoA (Table 2). Protocol 
waivers or exemptions are not allowed. 
• 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. 
• 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 lipids, other laboratory and PD biomarker tests, sample collection for PK 
and 
, prior to administration of study drug. 
• Immediate safety concerns should be discussed with study Medical Monitor 
immediately upon occurrence or awareness to determine if the subject should continue 
or discontinue IP. 
• Adherence to the study design requirements, including those specified in the SoA 
(Table 2), is essential and required for study conduct. 
• 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 subject management 
and pre-screening, however screening and baseline assessments must be obtained by 
central laboratory and as defined by the protocol. 
• 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/contract research organization (CRO) immediately upon 
occurrence or awareness to determine potential impact on study subject/s or study 
conduct. 
8.1. 
Efficacy Assessments 
Fasting blood samples will be collected at the time points specified in the SoA for the 
measurement of serum total lipids, lipoprotein profiles, and apolipoproteins. Efficacy assessments 
in this study are the PD and biomarker assessments, which are described in Section 8.6. 
8.2. 
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. 
Any clinically significant safety assessment findings, including physical examination, ECG, vital 
signs, laboratory assessments, and AEs that are considered to be related to COVID-19 infection 
should be documented as such in the AE term reported (e.g., COVID-related pneumonia).  
All safety analyses will be performed using the Safety Analysis Set (Section 9.4.5). Planned time 
points for all safety assessments are provided in the SoA (Table 2). 
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8.2.1. 
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). 
8.2.2. 
Vital Signs 
• Vital signs including body temperature, pulse rate, RR, and supine BP will be assessed. 
• Planned time points for vital signs assessment are provided in the SoA (Table 2). 
• Starting from randomization, BP and pulse will be measured in duplicate. The first 
measurement will be initiated before dosing with sitting/semi-erect or supine position 
and resting for at least 5 minutes prior to measurements. BP and pulse are to be 
repeated at least 1 minute apart. Additional vital signs measurement may be done if 
clinically indicated. 
8.2.3. 
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. 
8.2.4. 
Clinical Safety Laboratory Assessments 
• See Appendix 2 (Section 10.2) or the list of clinical laboratory tests to be performed 
and the SoA (Table 2) for the timing and frequency. Laboratory tests should be 
performed under fasting conditions (for at least 12 hours; it is recommended that 
subjects fast no more than 14 hours). Subjects should be instructed to attempt to remain 
consistent in how long they fast prior to blood draws for the duration of the study. 
• Fasting is defined as taking nothing by mouth except non-mineral water (and any 
essential medications). It is required that subjects abstain from consumption of 
alcoholic beverages for at least 48 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. 
• Retesting of laboratory tests is allowed when results are deemed spurious or 
incomplete. 
• All laboratory tests with values considered clinically significantly abnormal during 
participation in the study or within 28 days after the last dose of IP should be repeated 
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until the values return to normal or baseline or are no longer considered clinically 
significant by the Investigator or the Medical Monitor. 
• If such values do not return to normal/baseline within a period of time judged 
reasonable by the Investigator and remain clinically significant, the etiology should be 
identified and the Medical Monitor notified. 
• All protocol-required laboratory assessments, as defined in Appendix 2 (Section 10.2), 
must be conducted in accordance with the relevant study manual and the SoA 
(Table 2). 
• 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 Medical Monitor should 
be notified. 
8.3. 
Adverse Events and Serious Adverse Events 
The definitions of an AE or SAE can be found in Appendix 3 (Section 10.3). 
Adverse events 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 IP or study procedures, or that caused the subject 
to discontinue the study or IP (see Section 7). 
8.3.1. 
Time Period and Frequency for Collecting AE and SAE Information 
The timing and frequency of collection of all AEs and SAEs is specified in the SoA (Table 2). 
Table 4 summarizes the different reporting periods for AEs, SAEs and events to monitor. Events 
to monitor are defined in Section 10.3.3. 
Table 4: 
Adverse Event Reporting Periods 
Type of 
Event 
Adverse Event 
Serious Adverse Event 
Events to Monitor with 
Investigational Product 
Reporting 
period 
From consent until 
28 days after last dose of 
investigational product 
From consent until 28 days 
after the last dose of 
investigational product  
From consent until 28 days 
after the last dose of 
investigational product 
Reporting 
Timelines to 
89bio  
Entered into the clinical 
database on an ongoing 
basis 
Within 24 hours  
Within 24 hours 
All SAEs will be recorded and reported to 89bio or designee immediately and under no 
circumstance should this exceed 24 hours, as indicated in Appendix 3 (Section 10.3). The 
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Investigator will submit any updated SAE data to 89bio 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 IP or study participation, the Investigator must promptly notify 89bio or designee. 
8.3.2. 
Method of Detecting AEs and SAEs 
The method of recording, evaluating, and assessing causality of AEs and SAEs and the procedures 
for completing and transmitting SAE reports are provided in Appendix 3 (Section 10.3). 
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended and 
non-leading verbal questioning of the subject is the preferred method to inquire about 
AE occurrences. 
8.3.3. 
Follow-up of AEs and SAEs 
After the initial AE/SAE report, the Investigator is required to proactively follow each subject at 
subsequent visits/contacts. All SAEs will be followed until resolution, stabilization, the event is 
otherwise explained, or the subject is lost to follow-up (as defined in Section 7.4). Further 
information on follow-up procedures is provided in Appendix 3 (Section 10.3). 
8.3.4. 
Regulatory Reporting Requirements for SAEs 
• Prompt notification by the Investigator to 89bio or designee of an SAE is essential so 
that legal obligations and ethical responsibilities regarding the safety of subjects and 
the safety of the IP under clinical investigation are met. 
• 89bio has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of the IP under clinical investigation. 89bio will 
comply with country-specific regulatory requirements relating to safety reporting to the 
regulatory authority, IRBs/IECs, and Investigators. 
• Investigator safety reports must be prepared for suspected unexpected serious adverse 
reactions (SUSARs) according to local regulatory requirements and 89bio 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 89bio will review 
and then file it along with the IB and will notify the IRB/IEC, if appropriate according 
to local requirements. 
8.3.5. 
Pregnancy 
• Details of all pregnancies in female subjects and in female partners of male subjects 
will be collected after the start of IP and until 90 days after last dose of IP. 
• If a pregnancy is reported, the Investigator should inform 89bio within 24 hours of 
learning of the pregnancy and should follow the procedures outlined in Appendix 4 
(Section 10.4). 
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• Abnormal pregnancy outcomes (e.g., spontaneous abortion, fetal death, stillbirth, 
congenital anomalies, ectopic pregnancy) are considered SAEs. 
8.4. 
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. 
89bio does not recommend specific treatment for an overdose. 
In the event of an overdose, the Investigator should: 
1. Contact the Medical Monitor immediately. 
2. Monitor the subject’s vital signs and institute supportive measures as indicated by the 
subject’s clinical state. 
3. 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). 
4. Document the quantity of the excess dose as well as the incidence of the overdose in the 
eCRF. 
8.5. 
Pharmacokinetics 
Blood samples for analysis of BIO89-100 serum levels will be collected at the timepoints 
designated in the SoA (Table 2). 
Blood samples will be processed for collection of serum fractions for determination of BIO89-100 
serum concentrations. BIO89-100 metabolites may be analyzed. 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. 
When appropriate, pharmacokinetic parameters calculated by non-compartmental methods from 
the BIO89-100 serum concentration data will include: 
• Cmax 
• AUC0-tau 
• tmax 
• t½ 
Subjects may choose to participate in the Intensive PK scheme. All subjects will be included in the 
Trough PK scheme. Table 5 details sampling times for the Intensive PK scheme. For subjects in 
the Trough PK scheme, blood samples will be taken within 4 hours prior to dosing on specified 
dosing days or during EOT visit 
. 
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Table 5: 
Intensive PK Scheme for Sample Collection  
Note: Where the Intensive PK schedule overlaps with the Trough PK schedule, only 1 PK 
measurement should be taken at those timepoints. 
Additional PK parameters may be calculated if deemed appropriate. 
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8.6. 
Pharmacodynamics and Biomarkers 
8.6.1. 
Pharmacodynamics 
The following biomarkers/PD parameters will be evaluated at timepoints designated in the SoA 
(Table 2). 
Laboratory parameters: 
• TG 
• 
 
• VLDL-C 
• VLDL-TG 
• LDL-C 
• HDL-C 
• Non-HDL-C 
• ApoB 
• RLP-C 
• 
 
• 
 
• 
 
• 
 
• 
  
• ALT 
• AST 
• Adiponectin 
• hsCRP 
• 
 
• 
 
• 
 
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 89bio, 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. 
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Other parameters include: 
• Imaging measures (Section 8.6.3) 
− MRI-PDFF  
• Anthropomorphic measurements 
− Body weight 
− BMI 
− 
 
− 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
8.6.3. 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction  
MRI-PDFF is a non-invasive, quantitative, and accurate measure of liver fat content 
(imaging-based biomarker) to assess treatment response in clinical studies (Caussy, et al. 2018). 
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This technology enables post-processing of MRI data into a parametric map of Proton Density Fat 
Fraction (PDFF; Antaros Medical, Sweden) to provide accurate and quantitative measures of liver 
fat.  
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 (refer to the Imaging Manual for more 
information). The liver fat value (PDFF) is the mean value of all voxels in the identified volume of 
interest.  
After a consenting subject’s 
, the site should preemptively schedule the baseline MRI-PDFF to 
occur within the following projected visit windows to ensure an MRI appointment is available to 
be performed after the 
 TG result ≥350 mg/dL is known. For the main cohort, MRI-PDFF is 
optional and should occur prior to 
 (The study aims to enroll ~33% of subjects 
[~30 subjects total] in the main cohort with a baseline MRI-PDFF ≥6.0%.) For the fibrate 
expansion cohort, MRI-PDFF of ≥6.0% is required for enrollment and should be performed 
between 
 The follow-up MRI at 
 should only be 
performed if the baseline liver MRI-PDFF value is ≥6.0%. MRI-PDFF is mandatory in the fibrate 
expansion cohort. 
 
 
8.6.4. 
Exploratory Analyses 
 
 
 
 
 
 
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9. 
STATISTICAL CONSIDERATIONS 
9.1. 
Statistical Hypotheses 
The study hypothesis is that BIO89-100 can significantly improve efficacy parameters and is well 
tolerated for subjects with SHTG. 
The hypotheses for the primary endpoint are: 
• Null hypothesis: treatment with BIO89-100 and placebo result in the same percentage 
change in serum TG from baseline to Week 8 
• Alternative hypothesis: treatment with BIO89-100 and placebo result in significantly 
different percentage changes in serum TG from baseline to Week 8 
The null hypotheses will be tested at significance level of 0.05 (2-sided) for each BIO89-100 
treatment group versus placebo. 
9.2. 
Sample Size Determination 
The effect of BIO89-100 on the percentage change from baseline in TG is estimated based on 
prior clinical studies evaluating BIO89-100 and other TG lowering agents: 
 
 
 
 
 
 
 
 
 
Based on these results, it is estimated that the SD of the percent change in TG at Week 8 is ~40%. 
With 18 subjects in the placebo group and each BIO89-100 dose group, there would be at least 
86% power to detect a 45% difference in TG between each BIO89-100 dose group and placebo at 
the 2-sided alpha level of 0.05, assuming 50% reduction in BIO89-100 dose group and 5% in the 
placebo group.  
Fatty liver has been shown to be significantly associated with moderately severe or severe acute 
pancreatitis (Yoon, et al. 2017). Hence, another focus of this study is to assess the BIO89-100 
effect on reducing liver fat as assessed by MRI-PDFF in the SHTG population, with or without 
concurrent stable fibrate therapy. In Study BIO89-100-002 (Part 1), BIO89-100 (27 mg QW) 
reduced liver fat as assessed by MRI-PDFF up to 60%. In the main cohort not on concomitant 
fibrate therapy, a sample size of ~30 subjects (e.g., ~33% of the main cohort total sample size, 
~6 placebo vs ~24 pooled BIO89-100) is estimated to provide ~89% power to detect a difference 
of at least 50% mean percentage reduction in MRI-PDFF at Week 8 compared to placebo, using a 
significance level of 0.05 (2-sided). These calculations are based on an assumption that the 
percentage change from baseline in MRI-PDFF is normally distributed with a standard deviation 
of no greater than 30%, as estimated from data in Study BIO89-100-002. In the fibrate expansion 
cohort, assuming a difference of 50% mean percentage reduction in MRI-PDFF at Week 8 
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compared to placebo, a standard deviation of no greater than 30% under normal assumption, and a 
significance level of 0.05 (2-sided), the planned sample size of 18 subjects per group will have 
>99% power to detect such a mean difference. 
This sample size assessment incorporates approximately 10% of subjects lost to follow-up. 
9.3. 
Populations for Analyses 
The following populations are defined for the main and fibrate expansion cohorts. 
Population 
Description 
Screened Analysis 
Set 
All subjects who signed informed consent and have undergone screening. 
Randomized 
Analysis Set 
All subjects in screened analysis set who are assigned a randomization number in 
the study. 
Full Analysis Set 
All randomized subjects who received at least 1 dose of IP, have a baseline, and at 
least 1 post-baseline efficacy measurements. 
Safety Analysis Set 
All subjects who received at least 1 dose of IP. 
Intensive PK 
Analysis Set 
All subjects in the Safety Analysis Set, who opted to participate in the Intensive 
PK scheme, and have at least 1 on-study PK measurement. 
MRI-PDFF 
Analysis Set  
All subjects in the Full Analysis Set who have MRI-PDFF at baseline and a post-
baseline assessment. 
9.4. 
Statistical Analyses 
The analyses presented here represent an outline of the intended methodology. Changes, additions, 
and further details about the analyses will be described in the statistical analysis plan (SAP), as 
applicable. The 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.  
Subjects with dose interruptions affected by the COVID-19 pandemic will be noted in the listings. 
The impact on COVID-19 related study interruption may be considered, as appropriate. 
9.4.1. 
General Considerations 
In general, summaries and analyses will be presented by dose/treatment group and by cohort. 
Descriptive statistics will be presented for demographics and baseline characteristics, safety 
endpoints, and PK and PD parameters. Continuous variables will be summarized by number of 
subjects and mean, SD/standard error, median, minimum, and maximum values. Categorical 
variables will be summarized by number and percentage of subjects. 
The analyses will be performed separately for the main cohort and the fibrate expansion cohort.  
All primary, secondary, and exploratory efficacy endpoints will be assessed in the Full Analysis 
Set. All safety assessments will be performed on the Safety Analysis Set. PK endpoints will be 
assessed in the Intensive PK Analysis Set as applicable. Endpoints of MRI-PDFF will be assessed 
in MRI-PDFF Analysis Set. 
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All statistical tests will be 2-sided and tested at a statistically significant level of 0.05 without 
adjustment for multiplicity. Confidence intervals will be 2-sided 95%, unless stated otherwise.  
9.4.2. 
Primary Endpoint(s) 
The primary efficacy analysis of the primary endpoint, percentage change in serum TGs from 
baseline to Week 8, will be performed using the Full Analysis Set and observed data, and an 
analysis of covariance model with treatment and background therapy as factors and baseline TG as 
a covariate. Baseline TG will be deﬁned as the average of 
 and the preceding 
lipid-qualifying visit 
. For the TG at Week 8, it will be deﬁned as 
the average of measurements at 
 and 
 If there are significant 
deviations from the normality assumption of the primary endpoint, the alternative non-parametric 
analysis will be performed using the median of the treatment differences and Hodges-Lehmann 2-
tailed 95% confidence interval, with treatment comparison using the Wilcoxon rank-sum test.  
Comparison of change from baseline to Week 8 in TG will be analyzed using the same analysis 
method for the primary endpoint. 
The following sensitivity analysis will be conducted: the percentage change from baseline in TG at 
each post-baseline study visit be analyzed using a Mixed Effects Model with Repeated Measures 
(model) where the treatment group, time, treatment by time interaction, and the randomization 
stratification factor of background therapy will be included in the model as fixed effects; subjects 
will be a random effect; the baseline TG value will be included as a covariate. No imputation for 
missing data will be made for this sensitivity analysis. The unstructured covariance model will be 
used. The treatment contrast for the analysis at each study visit, including 
, will 
be estimated by the model. 
9.4.3. 
Secondary Endpoint(s) 
The proportion of subjects with TG <500 mg/dL at Week 8 will be summarized by treatment 
group and analyzed using Cochran Mantel Haenszel (CMH) method stratified by the 
2 stratification factors. To examine whether the response is consistent over a range of response 
thresholds, responder analysis with other response thresholds will be considered.  
As sensitivity analyses of the responder of TG <500 mg/dL at Week 8, comparison between 
BIO89-100 and placebo groups of proportion of subjects with ≥50% reduction from baseline at 
Week 8 in TG will be analyzed using the CMH test stratified by the 2 stratification factors. To 
examine whether the response is consistent over a range of response thresholds, responder analysis 
with other response thresholds will be considered. Cumulative distribution function of both 
percentage changes from baseline and changes from baseline for TG at Week 8 by treatment group 
will be summarized descriptively and graphically. 
The analysis methods for the primary endpoint will be used to analyze other similar secondary 
efficacy endpoints. 
All serum concentrations of BIO89-100 will be listed. Pharmacokinetic parameters in the 
Intensive PK group will be determined where possible from the serum concentrations of 
BIO89-100. Further details on PK analysis will be provided in the SAP. 
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9.4.4. 
Exploratory Endpoints 
 
9.4.5. 
Safety Analyses 
Treatment duration and IP received will be summarized by treatment group. 
Subject incidence of TEAEs will be tabulated by Medical Dictionary for Regulatory Activities 
(MedDRA) system organ class, MedDRA preferred term, and treatment group. All TEAEs, all 
treatment-emergent related AEs, all TESAEs, and all treatment-emergent serious related AEs will 
be summarized. Laboratory tests, vital signs, and ECG measures will be summarized by visits and 
by treatment group. 
 
 
9.5. 
Interim Analyses  
No efficacy interim analysis will be performed. 
9.6. 
Data Monitoring Committee 
There is no data monitoring committee for this study. 
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10. 
SUPPORTING DOCUMENTATION AND OPERATIONAL 
CONSIDERATIONS 
10.1. 
Appendix 1: Regulatory, Ethical, and Study Oversight Considerations 
10.1.1. 
Regulatory and Ethical Considerations 
• This study will be conducted in accordance with the protocol and with the following: 
− Consensus ethical principles derived from international guidelines including the 
Declaration of Helsinki and Council for International Organizations of Medical 
Sciences International Ethical Guidelines 
− Applicable ICH GCP Guidelines 
− Applicable laws and regulations 
• The protocol, protocol amendments, ICF, IB, 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 CFR, ICH guidelines, the IRB/IEC, European regulation 
536/2014 for clinical studies (if applicable), and all other applicable local 
regulations 
10.1.2. 
Financial Disclosure 
Investigators and sub-investigators will provide 89bio with sufficient, accurate financial 
information as requested to allow 89bio 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. 
10.1.3. 
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. 
10.1.4. 
Data Protection 
• Subjects will be assigned a unique identifier by 89bio. Any subject records or datasets 
that are transferred to 89bio 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 
89bio in accordance with local data protection law. The level of disclosure must also be 
explained to the subject who will be required to give consent for their data to be used 
as described in the informed consent. 
• The subject must be informed that his/her medical records may be examined by 
Clinical Quality Assurance auditors or other authorized personnel appointed by 89bio, 
by appropriate IRB/IEC members, and by inspectors from regulatory authorities. 
10.1.5. 
Dissemination of Clinical Study Data 
A clinical study report will be developed by 89bio at completion of data analysis. This report will 
be a clinical and statistical integrated report, according to the ICH E3 guidelines. 
89bio will register the study and post-study results regardless of outcome on a publicly accessible 
website in accordance with the applicable laws and regulations. 
10.1.6. 
Data Quality Assurance 
• All subject data relating to the study will be recorded on the eCRFs unless transmitted 
to 89bio 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. 
• The Investigator must permit study-related monitoring, audits, IRB/IEC review, and 
regulatory agency inspections and provide direct access to source data documents. 
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• Monitoring details describing strategy, methods, responsibilities and requirements, 
including handling of non-compliance issues and monitoring techniques (central, 
remote, or on-site monitoring) are provided in the Monitoring Plan. 
• 89bio or designee is responsible for the data management of this study including 
quality checking of the data. 
• 89bio assumes accountability for actions delegated to other individuals (e.g., CROs). 
• 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. 
• 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 89bio. No records may be transferred to another location or party without 
written notification to 89bio, whether within the retention period or thereafter. 
10.1.7. 
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. 
10.1.8. 
Study and Site Start and Closure 
89bio reserves the right to close a study site or terminate the study at any time for any reason. 
Study sites will be closed upon study 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 89bio in advance of the intended termination. 
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Reasons for the early closure of a study site by 89bio 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, 89bio’s procedures, or GCP guidelines 
• Inadequate recruitment of subjects by the Investigator 
• Discontinuation of further IP development 
• Debarment  
If the study is prematurely terminated or suspended, 89bio shall promptly inform the Investigators, 
the IECs/IRBs, the regulatory authorities, and any CRO(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. 
10.1.9. 
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 89bio before 
submission and only present or publish information after approval by 89bio. This 
allows 89bio to protect proprietary information and to provide comments. 
• 89bio will comply with the requirements for publication of study results. In accordance 
with standard editorial and ethical practice, 89bio 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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10.2. 
Appendix 2: Clinical Laboratory Tests 
• The clinical laboratory tests detailed in Table 6 will be performed by a central 
laboratory at timing/frequency detailed in the SoA (Table 2). Laboratory tests will be 
performed under fasting conditions (for at least 12 hours; it is recommended that 
subjects fast no more than 14 hours). Subjects should be instructed to attempt to remain 
consistent in how long they fast prior to blood draws for the duration of the study. 
• Protocol-specific requirements for inclusion or exclusion of subjects are detailed in 
Section 5 of the protocol.  
• Additional tests may be performed at any time during the study as determined 
necessary by the Investigator or required by local regulations. 
• Investigators must document their review of each laboratory safety report. 
Table 6: 
Protocol-Required Laboratory Assessments 
Hematology 
 
WBC with differential (Total 
Neutrophils, Lymphocytes, Monocytes, 
Eosinophils, Basophils – absolute and 
%) 
RBC count 
RBC Indices: 
MCV, MCH, MCHC, RDW, Reticulocyte count 
Hemoglobin 
Hematocrit 
Platelet count 
Coagulation panel, including PT, INR, aPTT  
Lipid Assessment 
 
 
non-HDL-C* 
ApoB* 
RLP-C* 
 
 
HDL-C* 
TG* 
 
VLDL-C* 
LDL-C* 
VLDL-TG* 
 
 
Clinical Chemistries 
 
ALP 
Fasting plasma glucose 
ALT 
GGT 
AST 
Lactate dehydrogenase 
Albumin 
Magnesium 
Bicarbonate 
Phosphorus 
BUN 
Potassium 
Calcium 
Sodium 
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Creatinine, eGFR  
Total bilirubin, Indirect/direct bilirubin 
CK 
Total protein 
Chloride 
Uric acid 
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 are abnormal or at the discretion of 
the PI or delegate. 
Other Study-Specific Laboratory 
Assessments 
 
BIO89-100 (to be evaluated by 
bioanalytical laboratory) 
Thyroid function assessment (TSH, 
free T4, and free T3) 
 
 
 
Serum hCG pregnancy test 
Urine hCG pregnancy test# 
FSH level 
Urine drug screen including 
amphetamines, barbiturates, cocaine 
metabolites, opiates, benzodiazepines, 
cannabinoids and phencyclidine 
 
Serology – HIV antibody, hepatitis B 
surface antigen (HBsAg, and HCV 
antibody). To determine current 
infection status, if HBsAg positive HBV 
DNA will be measured, if HCV antibody 
positive, HCV RNA will be measured, 
and if HIV antibody positive will be 
differentiated by HIV 1 or 2 and HIV 
RNA will be measured. 
24-hour urine for cortisol and creatinine 
 
 
 
Insulin 
hsCRP 
 
 
Serum adiponectin, total 
 
 
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* Post-baseline lipid panels to be blinded 
# Tests can be performed locally with kits provided by the central laboratory 
Abbreviations: ALP = alkaline phosphatase; ALT = alanine aminotransferase; 
; 
ApoB = apolipoprotein B100; 
; aPTT = activated partial thromboplastin time; 
AST = aspartate aminotransferase; 
; BUN = blood urea nitrogen; CK = creatine 
kinase; 
; DNA = deoxyribonucleic acid; eGFR = estimated 
glomerular filtration rate using CKD-EPI (Chronic Kidney Disease Epidemiology Collaboration) equation; 
; FSH = follicle-stimulating hormone; GGT = gamma-glutamyl transferase; HBsAg = Hepatitis 
B surface antigen; 
; HBV = Hepatitis B virus; hCG = human chorionic gonadotropin; HCV 
= Hepatitis C Virus; HDL-C = high-density lipoprotein-cholesterol; HIV = human immunodeficiency virus; 
 
; INR = international normalized ratio; LDL-C = lipoprotein-cholesterol; 
 
; MCH = mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; PT = 
prothrombin time; PINP = n-terminal propeptide of type I procollagen; MCV = Mean Corpuscular Volume; Non-
HDL-C = non-high-density lipoprotein-cholesterol; RBC = red blood cell; RDW = Red Cell Distribution Width; RLP-
C = remnant lipoprotein-cholesterol; RNA = ribonucleic acid; T3 = triiodothyronine; T4 = thyroxine; 
 
; TG = triglycerides; TSH = thyroid-stimulating hormone; VLDL-C = very low-density lipoprotein 
cholesterol; VLDL-TG = very low-density lipoprotein triglycerides; WBC = white blood cell. 
10.3. 
Appendix 3: Adverse Events: Definitions and Procedures for 
Recording, Evaluating, Follow-up, and Reporting 
10.3.1. 
Definition of AE 
AE Definition 
• 
An AE is any untoward medical occurrence in a clinical study subject, temporally associated with 
the use of IP, whether or not considered related to the IP. 
• 
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 
IP. 
 
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 are 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 IP 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 IP 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. 
 
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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. 
10.3.2. 
Definition of Suspected and Unsuspected Adverse Reaction 
Suspected adverse reactions are defined as: 
• 
Any AE for which there is a reasonable possibility that the IP caused the AE. For the purposes of 
89bio regulatory safety reporting, “reasonable possibility” means there is evidence to suggest a 
causal relationship between the IP and the AE. A suspected adverse reaction implies a lesser degree 
of certainty about causality than adverse reaction, which means any AE caused by an IP 
Unexpected AEs are defined as: 
• 
AE which is not listed in the IB of the IP or is not listed at the specificity or severity that has been 
observed 
10.3.3. 
Definition of Events to Monitor 
89bio-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 89bio 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  
10.3.4. 
Definition of SAE 
If an event is not an AE per definition in Section 10.3.1, 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 
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An SAE is defined as any untoward medical occurrence that, at any dose: 
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 (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 1 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. 
10.3.5. 
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/paper case report 
form (CRF) as appropriate. 
• 
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 and paper CRF as appropriate. 
• 
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. 
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AE and SAE Recording 
• 
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 non-invasive 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. 
 
Assessment of Causality 
The Investigator is obligated to assess the relationship between IP 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 IP administration will be considered and 
investigated. 
• 
The Investigator will also consult the 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 makes 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 1 of the criteria used when determining regulatory reporting 
requirements. 
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Assessment of Causality 
Causality Categories: 
• 
UNRELATED – This relationship suggests that there is no association between the IP and the 
reported event. 
• 
UNLIKELY RELATED – The relationship suggests that the event is more likely due to other 
causes than IP, however the relationship to the IP cannot be ruled out. 
• 
POSSIBLY RELATED – This relationship is based on evidence suggesting a causal relationship 
between the IP 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 IP 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 IP 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. 
 
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 and paper CRF as 
appropriate. 
• 
The Investigator will submit any updated SAE data to 89bio within 24 hours of receipt of the 
information. 
10.3.6. 
Reporting of SAEs 
Reporting of SAEs will be done using a paper CRF. 
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. 
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SAE Reporting to the Medical Monitor via Paper CRF 
• 
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. 
10.4. 
Appendix 4: Contraceptive Guidance and Collection of Pregnancy 
Information 
Definitions: 
WOCBP 
A woman is considered fertile following menarche and until becoming postmenopausal 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 IP, additional evaluation should be considered. 
Women in the Following Categories Are Not Considered WOCBP 
1. Premenarchal 
2. Premenopausal female with 1 of the following: 
• Documented hysterectomy 
• Documented bilateral salpingectomy  
• Documented 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. 
• Note: Documentation can come from the site personnel’s review of the subject’s 
medical records or medical examination. An ultrasound to document absence of a 
uterus is considered acceptable documentation of non-childbearing potential. 
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 1 FSH measurement is required. 
• Females on HRT and whose menopausal status is in doubt will be required to use 1 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. 
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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 (with both male/female partners) 
during the study and for 30 days or 5 half-lives (whichever is longer) of IP. 
Double contraception is defined as a condom with 1 of any of the following forms of birth control: 
• Birth control pills (The Pill) 
• Depot or injectable birth control 
• IUD 
• Birth Control Patch (e.g., Ortho Evra) 
• NuvaRing® 
• Documented evidence of tubal ligation prior to screening visit for women or history of 
vasectomy for men 
Subjects must not donate sperm or oocytes during the study and for 30 days or 5 half-lives 
(whichever is longer) after last dose of IP. 
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 IP 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 IP. 
• 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 89bio 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 89bio. 
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 
• 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 89bio within 24 hours of learning of a subject’s 
pregnancy. This applies only to female subjects who receive IP. 
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• 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 89bio. 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 IP by 
the Investigator will be reported to 89bio as described in Section 8.3.4. 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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10.5. 
Appendix 5: Abbreviations 
Abbreviation Term 
Description 
~ 
Approximately 
ABV 
Alcohol By Volume 
 
 
 
 
ADL 
Activities of Daily Living 
AE 
Adverse Event 
ALP 
Alkaline Phosphatase 
ALT 
Alanine Aminotransferase (SGPT) 
AP 
Acute Pancreatitis 
Apo 
Apolipoprotein 
 
 
ApoB 
Apolipoprotein B100 
 
 
 
 
aPTT 
Activated Partial Thromboplastin Time 
AST 
Aspartate Aminotransferase (SGOT) 
AUC 
Area Under the Curve 
AUC0-tau 
Area Under the Curve during the Dosing Interval 
 
 
BMI 
Body Mass Index 
BP 
Blood Pressure 
BUN 
Blood Urea Nitrogen 
CBC 
Complete Blood Count 
CK 
Creatine Kinase 
CKD-EPI 
Chronic Kidney Disease Epidemiology Collaboration 
Cmax 
Maximum Concentration 
CMH 
Cochran Mantel Haenszel 
COVID-19 
Coronavirus Disease 2019 
CRF 
Case Report Form 
CRO 
Contract Research Organization 
CTCAE 
Common Terminology Criteria for Adverse Events 
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Abbreviation Term 
Description 
 
 
CVD 
Cardiovascular Disease 
CYP3A 
Cytochrome P450 3A 
DHA 
Docosahexaenoic acid 
DILI 
Drug-induced Liver Injury 
DNA 
Deoxyribonucleic Acid 
ECG 
Electrocardiogram 
eCRF 
Electronic Case Report Form 
eGFR 
Estimated Glomerular Filtration Rate 
EOS 
End of Study 
EOT 
End of Treatment 
EPA 
Eicosapentaenoic acid 
FDA 
Food and Drug Administration 
 
 
FGF21 
Fibroblast Growth Factor 21 
FGFR 
Fibroblast Growth Factor Receptor 
FSH 
Follicle-Stimulating Hormone 
GCP 
Good Clinical Practice 
GGT 
Gamma-Glutamyl Transferase 
GI 
Gastrointestinal 
 
 
HBV 
Hepatitis B Virus 
HBsAg 
Hepatitis B Surface Antigen 
HCG 
Human Chorionic Gonadotropin 
HCV 
Hepatitis C Virus 
HDL-C 
High-Density Lipoprotein Cholesterol 
HHP 
Home health provider 
HIPAA 
Health Insurance Portability and Accountability Act 
HIV 
Human Immunodeficiency Virus 
HRT 
Hormone replacement therapy 
hsCRP 
High-Sensitivity C-reactive Protein 
IB 
Investigator Brochure 
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Abbreviation Term 
Description 
ICF 
Informed Consent Form 
IEC 
Independent Ethics Committee 
ICH 
International Council for Harmonisation 
 
 
INR 
International Normalized Ratio 
IP 
Investigational Product 
IRB 
Institutional Review Board 
ISR 
Injection Site Reaction 
IUD 
Intrauterine Device 
IVRS 
Interactive Voice Response System 
IWRS 
Interactive Web Response System 
LDL 
Low-Density Lipoprotein 
LDL-C 
Low-Density Lipoprotein Cholesterol 
 
 
MAD 
Multiple Ascending Dose 
MCV 
Mean Corpuscular Volume 
MCH 
Mean Corpuscular Hemoglobin 
MCHC 
Mean Corpuscular Hemoglobin Concentration 
MedDRA 
Medical Dictionary for Regulatory Activities 
MRI 
Magnetic Resonance Imaging 
MRI-PDFF 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction 
NAb 
Neutralizing Antibody 
NAb- 
Neutralizing Antibody Negative 
NAFLD 
Non-Alcoholic Fatty Liver Disease 
NASH 
Non-Alcoholic Steatohepatitis 
NOAEL 
No Observed Adverse Effect Level 
PCSK9 
Proprotein Convertase Subtilisin/Kexin Type 9 
PD 
Pharmacodynamic 
PDFF 
Proton Density Fat Fraction 
PEG 
Polyethylene Glycol 
PI 
Principal Investigator 
 
 
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Abbreviation Term 
Description 
PK 
Pharmacokinetic 
PT 
Prothrombin Time 
QW 
Every Week 
Q2W 
Every 2 weeks 
QTcF 
Fridericia’s-corrected QT interval 
RBC 
Red Blood Cell 
RDW 
Red Cell Distribution Width 
RLP-C 
Remnant Lipoprotein Cholesterol 
RNA 
Ribonucleic Acid 
RR 
Respiratory Rate 
SAD 
Single Ascending Dose 
SAE 
Serious Adverse Events 
SAP 
Statistical Analysis Plan 
SC 
Subcutaneous 
SD 
Standard Deviation 
SHTG 
Severe Hypertriglyceridemia 
SoA 
Schedule of Assessments 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
t½ 
Half-life 
T1DM 
Type 1 Diabetes Mellitus 
T2DM 
Type 2 Diabetes Mellitus 
T3 
Triiodothyronine 
T4 
Thyroxine 
 
 
TEAE 
Treatment-Emergent Adverse Event 
TESAE 
Treatment-Emergent Serious Adverse Events 
TG 
Triglycerides 
tmax 
Time to Maximum Concentration 
TSH 
Thyroid-Stimulating Hormone 
ULN 
Upper Limit of Normal 
V 
Visit 
VLDL-C 
Very Low-Density Lipoprotein Cholesterol 
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Abbreviation Term 
Description 
VLDL-TG 
Very Low-Density Lipoprotein Triglycerides 
WBC 
White Blood Cell 
WOCBP 
Women of Childbearing Potential 
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Brisson D, Methot J, Tremblay K, et al. Comparison of the efficacy of fibrates on 
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Gaich G, Chien JY, Fu H, et al. The effects of LY2405319, an FGF21 analog, in obese human 
subjects with type 2 diabetes. Cell Metab. 2013;18(3): 333-340. 
Galman C, Lundasen T, Kharitonenkov A, et al. The circulating metabolic regulator FGF21 is 
induced by prolonged fasting and PPARα activation in man. Cell Metab. 2008;8(2):169-74. 
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Grundy, S. M., Stone, N. J., Bailey, A. L., Beam, C., Birtcher, K. K., Blumenthal, R. S., et al 
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Zhang J, Zhao Y, Xu C, et al. Association between serum free fatty acid levels and non-alcoholic 
fatty liver disease: a cross-sectional study. Sci Rep. 2014;4. 
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Statistical Analysis Plan 
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Statistical Analysis Plan 
A Phase 2, Randomized, Double-Blind, Placebo-
Controlled Study to Explore the Efficacy and 
Safety of BIO89-100 in Subjects with Severe 
Hypertriglyceridemia 
PROTOCOL NO. 
BIO89-100-221 Version 3.0 
Sponsor: 
89BIO 
CRO: 
 
Version:  
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Date: 
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TABLE OF CONTENTS 
TABLE OF CONTENTS ................................................................................................................................ 4 
LIST OF TABLES ......................................................................................................................................... 6 
LIST OF FIGURES ....................................................................................................................................... 6 
ABBREVIATIONS ....................................................................................................................................... 7 
1 
INTRODUCTION .............................................................................................................................. 9 
2 
STUDY OBJECTIVES .......................................................................................................................... 9 
2.1 
PRIMARY OBJECTIVE .............................................................................................................................. 9 
2.2 
SECONDARY OBJECTIVES ......................................................................................................................... 9 
2.3 
OTHER OBJECTIVES ................................................................................................................................ 9 
3 
STUDY DESIGN .............................................................................................................................. 10 
3.1 
STUDY POPULATION ............................................................................................................................. 10 
3.2 
DOSAGE AND ADMINISTRATION ............................................................................................................. 10 
3.3 
BLINDING AND RANDOMIZATION METHODS ............................................................................................. 11 
3.4 
STRATIFICATION FACTORS ...................................................................................................................... 12 
3.5 
SAMPLE SIZE CONSIDERATIONS .............................................................................................................. 12 
3.6 
INTERIM ANALYSIS ............................................................................................................................... 12 
3.7 
DATA MONITORING COMMITTEE ............................................................................................................ 12 
4 
STUDY ENDPOINTS AND COVARIATES ........................................................................................... 13 
4.1 
PRIMARY EFFICACY ENDPOINT ............................................................................................................... 13 
4.2 
SECONDARY EFFICACY ENDPOINTS .......................................................................................................... 13 
4.3 
OTHER EFFICACY ENDPOINTS ................................................................................................................. 13 
4.4 
SAFETY ENDPOINTS.............................................................................................................................. 14 
4.4.1 
Adverse Events ...................................................................................................................... 14 
4.4.2 
Clinical Safety Laboratories ................................................................................................... 14 
4.4.3 
Electrocardiogram, Vital Signs and Physical Examination .................................................... 14 
4.5 
PHARMACOKINETICS ENDPOINTS ............................................................................................................ 15 
4.7 
COVARIATES AND SUBGROUPS ............................................................................................................... 15 
5 
DEFINITIONS ................................................................................................................................. 16 
5.1 
BASELINE DEFINITION .......................................................................................................................... 16 
5.2 
ACTUAL TREATMENT GROUP ................................................................................................................. 17 
5.3 
STUDY DAY ........................................................................................................................................ 17 
5.4 
END OF STUDY .................................................................................................................................... 17 
5.5 
STUDY SCOPE ..................................................................................................................................... 17 
5.6 
PRE-TREATMENT MEDICATION AND PRE-TREATMENT PROCEDURES ............................................................... 18 
5.7 
BASELINE MEDICATION ......................................................................................................................... 18 
5.8 
CONCOMITANT MEDICATION AND CONCOMITANT PROCEDURES ................................................................... 18 
5.9 
ADVERSE EVENT AND SERIOUS ADVERSE EVENT ........................................................................................ 18 
5.10 
TREATMENT EMERGENT ADVERSE EVENT ............................................................................................ 19 
6 
ANALYSIS POPULATIONS ............................................................................................................... 19 
6.1 
HANDLING OF MISSING AND INCOMPLETE DATA ........................................................................................ 19 
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Statistical Analysis Plan 
Date: 03June2022 
 
Confidential 
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6.1.1 
Missing Dates........................................................................................................................ 19 
6.1.2 
Missing data for Efficacy Endpoints ...................................................................................... 21 
 
 
6.2 
ANALYSIS VISIT WINDOWS .................................................................................................................... 21 
6.3 
SOFTWARE ......................................................................................................................................... 22 
7 
STATISTICAL METHODS OF ANALYSES ............................................................................................ 22 
7.1 
GENERAL CONSIDERATIONS ................................................................................................................... 22 
7.2 
SUBJECT ACCOUNTABILITY ..................................................................................................................... 23 
7.3 
PROTOCOL DEVIATIONS ........................................................................................................................ 23 
7.4 
INVESTIGATIONAL PRODUCT ADMINISTRATION .......................................................................................... 24 
7.4.1 
Extent of Exposure and Treatment Compliance .................................................................... 24 
7.4.2 
Medical History ..................................................................................................................... 24 
7.4.3 
Demographics and Baseline Characteristics ......................................................................... 24 
7.4.4 
Prior, Baseline and Concomitant Medication/Procedure ...................................................... 25 
7.5 
EFFICACY ANALYSIS .............................................................................................................................. 26 
7.5.1 
Analysis of Primary Efficacy endpoints ................................................................................. 26 
 
 
7.5.2 
Analysis of Secondary Efficacy Endpoints ............................................................................. 27 
7.5.3 
Analysis of Other Efficacy Endpoints ..................................................................................... 27 
7.6 
SAFETY ANALYSIS................................................................................................................................. 28 
7.6.1 
Adverse Events ...................................................................................................................... 28 
7.6.2 
Clinical Laboratory Parameters ............................................................................................. 29 
7.6.3 
Vital Signs ............................................................................................................................. 30 
7.6.4 
Physical Examinations ........................................................................................................... 31 
7.6.5 
Electrocardiograms ............................................................................................................... 31 
7.7 
ANALYSIS OF SERUM CONCENTRATIONS OF BIO89-100 ............................................................................. 32 
7.8 
PHARMACOKINETICS ENDPOINTS ............................................................................................................ 32 
8 
LIST OF PLANNED TABLES, FIGURES ............................................................................................... 34 
9 
APPENDICES .................................................................................................................................. 35 
APPENDIX 1 SCHEDULE OF ACTIVITIES (SOA) ..................................................................................................... 35 
APPENDIX 2 SAS SAMPLE CODE ...................................................................................................................... 41 
 
 
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LIST OF TABLES 
TABLE 1. SUSPECTED LIVER INJURY CASES ACCORDING TO DRUG INDUCED LIVER INJURY (DILI) CRITERIA .............................. 14 
TABLE 2. TABLE HEADER FOR POOLED ANALYSIS ...................................................................................................... 17 
TABLE 3. TABLE HEADER FOR INDIVIDUAL DOSE GROUP ANALYSIS .............................................................................. 18 
TABLE 3-1. TABLE HEADER FOR ALL OTHER ANALYSIS ............................................................................................... 18 
TABLE 4. ANALYSIS POPULATION........................................................................................................................... 19 
TABLE 5. ANALYSIS VISIT WINDOW ........................................................................................................................ 22 
TABLE 6. BLOOD PRESSURE CATEGORY .................................................................................................................. 31 
 
LIST OF FIGURES 
FIGURE 1. STUDY SCHEMA: MAIN COHORT ............................................................................................................ 10 
FIGURE 2. STUDY SCHEMA: FIBRATE EXPANSION COHORT ......................................................................................... 11 
 
 
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Protocol No: BIO89-100-221 V3.0 
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Statistical Analysis Plan 
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ABBREVIATIONS 
Abbreviation 
Term 
 
 
AE 
Adverse Event 
ALP 
Alkaline Phosphatase 
ALT 
Alanine Aminotransferase (SGPT) 
 
 
ApoB 
Apolipoprotein B100 
 
 
aPTT 
Activated Partial Thromboplastin Time 
AST 
Aspartate Aminotransferase (SGOT) 
AUC 
Area Under the Curve 
AUC0-tau 
Area Under the Curve During The Dosing Interval 
 
 
BMI 
Body Mass Index 
BP 
Blood Pressure 
BUN 
Blood urea nitrogen 
CK 
Creatine Kinase 
Cmax 
Maximum Concentration 
CMH 
Cochran Mantel Haenszel 
CRF 
Case Report Form 
 
 
DNA 
Deoxyribonucleic Acid 
ECG 
Electrocardiogram 
eGFR 
Estimated Glomerular Filtration Rate 
EOS 
End of Study 
EOT 
End of Treatment 
 
 
FGF21 
Fibroblast Growth Factor 21 
FSH 
Follicle-Stimulating Hormone 
GGT 
Gamma-Glutamyl Transferase 
 
 
HBV 
Hepatitis B Virus 
HBsAg 
Hepatitis B Surface Antigen 
HCV 
Hepatitis C Virus 
HDL-C 
High Density Lipoprotein Cholesterol 
HIV 
Human Immunodeficiency Virus 
hsCRP 
High-Sensitivity C-reactive Protein 
 
 
INR 
International Normalized Ratio 
IVRS 
Interactive Voice Response System 
IWRS 
Interactive Web Response System 
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Protocol No: BIO89-100-221 V3.0 
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Statistical Analysis Plan 
Date: 03June2022 
 
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Abbreviation 
Term 
LDL 
Low Density Lipoprotein 
LDL-C 
Low Density Lipoprotein Cholesterol 
 
 
MCV 
Mean Corpuscular Volume 
MCH 
Mean Corpuscular Hemoglobin 
MCHC 
Mean Corpuscular Hemoglobin Concentration 
MedDRA 
Medical Dictionary for Regulatory Activities 
MRI-PDFF 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat 
Fraction 
OLE 
Open Label Extension 
PD 
Pharmacodynamic 
PE 
Physical Examination 
PEG 
Polyethylene Glycol 
PI 
Principal Investigator 
 
 
PK 
Pharmacokinetic 
PT 
Preferred Term 
QW 
Every Week 
Q2W 
Every 2 weeks 
QTcF 
Fridericia’s-corrected QT interval 
RBC 
Red blood cell 
RDW 
Red Cell Distribution Width 
RLP-C 
Remnant Lipoprotein Cholesterol 
RNA 
Ribonucleic Acid 
RR 
Respiratory Rate 
SAE 
Serious Adverse Events 
SC 
Subcutaneous 
SD 
Standard Deviation 
SHTG 
Severe Hypertriglyceridemia 
T½ 
Half-life 
 
 
TEAE 
Treatment-Emergent Adverse Event 
TESAE 
Treatment-Emergent Serious Adverse Events 
TG 
Triglycerides 
Tmax 
Time to Maximum Concentration 
TSH 
Thyroid Stimulating Hormone 
V 
Visit 
VLDL-C 
Very Low Density Lipoprotein Cholesterol 
VLDL-TG 
Very Low Density Lipoprotein Triglycerides 
WBC 
White Blood Cell 
 
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Protocol No: BIO89-100-221 V3.0 
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Statistical Analysis Plan 
Date: 03June2022 
 
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1 INTRODUCTION 
The purpose of this statistical analysis plan (SAP) is to describe the statistical methods, 
data derivations and data summaries that will be used to analyze and report results for 
Protocol BIO89-100-221 Version 3.0. 
The SAP will supersede the protocol in the event of any differences between the two 
documents in the plans for data analysis, and the protocol will be amended if appropriate. 
The SAP will be included as an appendix in the clinical study report. 
2 STUDY OBJECTIVES 
This Phase 2 study is designed to evaluate the efficacy, safety, and tolerability of different 
doses and dose regimens (every week [QW] or every 2 weeks [Q2W]), subcutaneous (SC) 
dosing of BIO89-100 compared to placebo in subjects with Severe Hypertriglyceridemia 
(SHTG). 
2.1 PRIMARY OBJECTIVE 
The primary objective is to determine the effect of BIO89-100 on serum Triglycerides (TG) 
levels in subjects with SHTG (TG ≥500 mg/dL). 
2.2 SECONDARY OBJECTIVES 
The secondary objectives of the study are: 
• To determine the effect of BIO89-100 on selected serum lipids and lipoproteins  
• To determine the effect of BIO89-100 on high sensitivity C-reactive protein 
(hsCRP) 
• To determine the effect of BIO89-100 on metabolic markers 
• To characterize BIO89-100 pharmacokinetics (PK) 
• To characterize BIO89-100 pharmacodynamic (PD) profile as assessed by 
Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction (MRI-
PDFF) 
2.3 OTHER OBJECTIVES 
The other objectives of the study are: 
• 
 
• 
 
The safety objectives of the study are: 
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• To evaluate the safety and tolerability of BIO89-100 
• To determine the effect of BIO89-100 on
 
• Other safety assessments 
3 STUDY DESIGN  
3.1 STUDY POPULATION 
Study BIO89-100-221 is a randomized, double-blind, placebo-controlled, Phase 2 study to 
evaluate the efficacy, safety, tolerability, PK and PD profiles, and 
 of 
BIO89-100 administered QW or Q2W SC for 8 weeks and followed by a 4-week follow-
up period.  
The main cohort will include approximately 90 subjects with SHTG without concurrent 
fibrate therapy and will consist of 5 treatment groups to compare 4 dose levels/regimens of 
BIO89-100 versus placebo (Figure 1). The fibrate expansion cohort will include 
approximately 36 subjects with SHTG on stable background fibrate therapy and with a 
baseline MRI-PDFF ≥6% and will consist of 2 treatment groups comparing one dose 
regimen of BIO89-100 versus placebo (Figure 2). 
The study includes Lifestyle Stabilization, TG Qualification, Treatment, and Follow-up 
periods. 
3.2 DOSAGE AND ADMINISTRATION 
Subjects enroll in the main cohort will be dosed with one of four doses of BIO89-100 (9 
mg QW, 18 mg QW, 27 mg QW, or 36 mg Q2W) or placebo. The subjects enroll in the 
fibrate expansion cohort will receive BIO89-100 27 mg or placebo QW. Intensive PK is 
planned for a subset of subjects (optional participation with no more than 45 subjects).  
 
Figure 1. Study Schema: Main Cohort  
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Statistical Analysis Plan 
Date: 03June2022 
 
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Figure 2. Study Schema: Fibrate Expansion Cohort 
3.3 BLINDING AND RANDOMIZATION METHODS 
Eligible subjects will be randomized in the order they are enrolled into the study. Subjects 
in the main cohort will be randomized in a 1:1:1:1:1 ratio to 1 of 4 doses of BIO89-100 (9 
mg QW, 18 mg QW, 27 mg QW, or 36 mg Q2W) or placebo. Subjects in the fibrate 
expansion cohort will be randomized in a 1:1 ratio to BIO89-100 27 mg or placebo QW.  
 
  
Randomization in the main cohort will be stratified by TG <750 mg/dL or ≥750 mg/dL 
(8.47 mmol/L) and by whether or not the subject is receiving background therapy (e.g., 
prescription fish oil and/or statins). 
 
 
 All subjects will be 
centrally assigned to randomized IP using an Interactive Voice/Web Response System 
(IVRS/IWRS). Before the study is initiated, the log-in information and directions for the 
IVRS/IWRS will be provided to each site.  
The study will be conducted under double-blind conditions. The subjects, PI , other study 
personnel involved with subject assessments, and 89bio will remain blinded to the actual 
treatment assignments of the subjects, post-baseline lipid panels (Triglycerides [TG], 
 
 
, Low Density Lipoprotein Cholesterol [LDL-C], High Density 
Lipoprotein Cholesterol [HDL-C], Non-high Density Lipoprotein Cholesterol [Non-HDL-
C], Very Low Density Lipoprotein Cholesterol [VLDL-C], Very Low Density Lipoprotein 
Triglycerides [VLDL-TG], 
, Apolipoprotein B100 [ApoB]), 
, Remnant Lipoprotein Cholesterol [RPL-C], 
 
, 
, and PK data. Blinded IP will be provided by a clinical supplies vendor and 
shipped to the study site. IP may be delivered to the subject’s home for home administration 
by an HHP. The SC administration of the blinded IP will be performed by trained study 
staff, and only blinded staff will be involved with subject assessments. 
 
 
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Protocol No: BIO89-100-221 V3.0 
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Statistical Analysis Plan 
Date: 03June2022 
 
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3.4 STRATIFICATION FACTORS 
Randomization in the main cohort will be stratified by (i) TG <750 mg/dL or ≥750 mg/dL 
(8.47 mmol/L) and (ii) whether the subject is receiving background therapy (e.g., 
prescription fish oil and/or statins) – Yes / No. 
 
 
3.5 SAMPLE SIZE CONSIDERATIONS 
The effect of BIO89-100 on the percentage change from baseline in TG is estimated based 
on prior clinical studies evaluating BIO89-100 and other triglyceride lowering agents. 
 
 
 
 
 
 
 
 
 
 
 
Based on these results, it is estimated that the SD of the percent change in TG at Week 8 is 
approximately 40%. With 18 subjects in the placebo group and each BIO89-100 active 
treatment group, there would be at least 86% power to detect a 45% difference in TG 
between each BIO89-100 active treatment group and placebo at the 2-sided alpha level of 
0.05, assuming 50% reduction in BIO89-100 dose group and 5% in the placebo group. This 
sample size assessment incorporates approximately 10% of subjects will be lost to follow-
up.  
Approximately 90 subjects are planned to be enrolled in the main cohort, with 18 subjects 
in each of the BIO89-100 active treatment groups (9 mg QW, 18 mg QW, 27 mg QW, or 
36 mg Q2W) and in the placebo group. The study aims to enroll ~33% of subjects (~30 
subjects total) in the main cohort with a baseline MRI-PDFF ≥ 6%. Approximately 36 
subjects are planned to be enrolled in the fibrate expansion cohort (18 in each treatment 
group) with all subjects having baseline MRI-PDF ≥ 6%. Subjects who terminate early in 
either cohort may be replaced at Sponsor’s discretion. 
3.6 INTERIM ANALYSIS 
No efficacy interim analysis is planned for this study.   
3.7 DATA MONITORING COMMITTEE 
No data monitoring committee is planned for this study. 
 
 
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Statistical Analysis Plan 
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4 STUDY ENDPOINTS AND COVARIATES 
4.1 PRIMARY EFFICACY ENDPOINT  
• Percentage change in serum TG from baseline to Week 8 
4.2 SECONDARY EFFICACY ENDPOINTS  
• The proportion of subjects who achieve TG < 500 mg/dL at Week 8  
• Percentage change in VLDL-C, LDL-C, non-HDL-C, HDL-C, VLDL-TG, ApoB, 
remnant lipoprotein cholesterol (RLP-C), hsCRP from baseline to Week 8 
• Percentage change in fasting plasma glucose, adiponectin, and body weight from 
baseline to Week 8 
• The serum BIO89-100 concentrations and PK parameters 
• Percentage change and change from baseline in hepatic steatosis using Magnetic 
Resonance Imaging – Whole Liver Proton Density Fat Fraction (MRI-PDFF)  
4.3 OTHER EFFICACY ENDPOINTS  
• 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
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• 
  
4.4 SAFETY ENDPOINTS 
4.4.1 Adverse Events 
The evaluation of adverse events (AE) will include only incidence of treatment-emergent 
AEs (TEAEs), defined as AEs with onset date and time after or identical to the date and 
time of the first dose of the investigational product until the last dose + 28 days  or 
continuing AEs that worsen the grade post IP. When onset time is missing or partial missing, 
AE occurs on the same day as first dose date are considered as TEAE.  The frequency of 
TEAEs, treatment-emergent serious AEs (TESAEs), and number of subjects who 
discontinue due to TEAEs will be summarized. 
4.4.2 Clinical Safety Laboratories 
The evaluation of clinical safety laboratories, including blood hematology, chemistry, 24-
hour urine cortisol, and urinalysis will be based on the observed values. Observed values 
and changes from baseline will be summarized for all post-baseline study visits. 
Incidence and shifts of laboratory abnormalities will be summarized. 
Changes in liver function tests (i.e., alkaline phosphatase (ALP), alanine aminotransferase 
(ALT), aspartate aminotransferase (AST), albumin and Gamma-Glutamyl Transferase 
(GGT)) from baseline to Week 8 will be summarized. A summary of suspected liver injury 
cases according to drug induced liver injury (DILI) criteria will also be presented (Table 
1). 
Table 1. Suspected liver injury cases according to drug induced liver injury (DILI) 
criteria 
Baseline Value of ALT/AST 
Criteria to discontinue investigational product  
>ULN and <2× ULN  
ALT or AST >5× baseline value  
>=2× ULN but <5× ULN  
ALT or AST >3× baseline value  
>=5× ULN 
ALT or AST >2× baseline value  
>ULN 
ALT or AST >2× baseline value and (BILI >2× ULN OR INR >0.2) 
Within Normal Range 
ALT or AST >8× ULN 
ALT or AST >5× ULN  
ALT or AST >3× ULN and (BILI >2× ULN or INR >1.5)  
 
 
 
4.4.3 Electrocardiogram, Vital Signs and Physical Examination  
The evaluation of electrocardiogram (ECG) and vital signs (including body temperature, 
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Statistical Analysis Plan 
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pulse rate, respiratory rate [RR], and supine Blood Pressure [BP]), will be summarized by 
visits and by treatment group.  
4.5 PHARMACOKINETICS ENDPOINTS 
PK parameters will be calculated by non-compartmental methods from the BIO89-100 
serum concentration data in Intensive PK subgroup, if the data are suitable.  
• 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) 
• Trough concentration 
Additional PK parameters may be calculated if also deemed appropriate. 
PK parameters will be calculated separately by designee of 89BIO. 
Serum concentration and PK parameters data will be analyzed using the Pharmacokinetics 
Analysis Set. 
 
 
 
 
 
 
 
 
 
4.7 COVARIATES AND SUBGROUPS 
• TG <750 mg/dL or ≥ 750 mg/dL 
• Background lipid medication (Yes, No) 
• Type of background lipid medication (use of statin (including combo therapy), use 
of fibrate therapy, use of prescription fish oil, Ezetimibe, and Bempedoic acid), 
provided there is sufficient number of subjects in the subgroup 
• Type 2 diabetes (Yes, No) 
• Age categories (21-<65, >=65). 
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Statistical Analysis Plan 
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• Sex (male, female) 
• Baseline weight <median and ≥ median 
• Region (US vs EU) 
• TG responder at Week 8 (responder, non-responder) 
• Use of background insulin medication (Yes/No) for insulin analysis only 
Note, if the percentage of subjects within a certain subgroup is less than 33% (i.e., less 
than 6 of 18 subjects) of the overall cohort, only descriptive analysis will be performed.  
More details of the analyses repeated in the above subgroups are described in Section 7.5.1. 
5 
DEFINITIONS  
5.1 BASELINE DEFINITION 
In general, baseline will be defined as the most recent non-missing measurement 
(scheduled or unscheduled) prior to the administration of the first dose of IP. For numerical 
measurements that have duplicates or triplicates (for the same visit), the arithmetic mean 
will be used. Some measurements have special definitions of baseline, which are defined 
below. 
The mean TG value will be used for stratification factor 
 
 
Baseline Triglyceride is defined as the average of 
 and the preceding two lipid-
qualifying visits collected in EDC. Fasting status is required for the TG values to be 
included. 
The definition of baseline of other lipid panel labs is the average of last two non-missing 
screening fasting values and 
 value (Pre-dose, fasting). 
The definition of baseline vital signs is the average of the last two non-missing screening 
values and 
 If there is Case Report Form (CRF) average value 
on duplicate measurements of BP and/or Pulse on 
, only the individual values 
will be included in the baseline derivation.  
The baseline definition for MRI for subjects of the main cohort is the 
 
value. The baseline definition for MRI for subjects of the fibrate expansion cohort is the 
value obtained between 
 
The baseline definition for safety numeric measurements (except for liver tests) is the 
average of last two non-missing screening values and 
  
The baseline definition of PK is the 
. The details will be described in a 
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separate document.  
5.2 ACTUAL TREATMENT GROUP 
 
 
 
 
 
5.3 STUDY DAY 
The day a subject receives their first dose of investigational product will be considered as 
  
If a subject is randomized but never dosed, study day will be calculated using the 
randomization date and randomization date will be set to 
 
If date of interest occurring on or after the relative date, relative day is calculated as: date 
of interest – relative date + 1, else if date of interest occurring prior the relative date, relative 
day is calculated as: date of interest – relative date. Data will be presented on listings in 
order of site number/subject number, assessment date and assessment (in order collected 
on CRF, unless specified otherwise). Dates will be presented in the format of YYYY-MM-
DD. 
For subjects with study interruptions due to COVID-19, post-interruption study day will 
be calculated as: (Index date) – (the date of 
 + 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. 
5.4 END OF STUDY 
A subject is considered to have completed the study if the subject has completed all phases 
of the study including the last scheduled procedure shown in Table 6. The EOS is defined 
as the date of the last visit of the last subject in the study. 
5.5 STUDY SCOPE  
For the efficacy analysis of the primary endpoint, results will be presented in two different 
formats as shown below:  
1. A comparison between the pooled BIO89-100 group and the pooled Placebo group. 
Subjects across all BIO89-100 dose groups will be pooled together to compare with all the 
placebo subjects.  The table header is shown below (Table 2). The Total group will be 
presented as appropriate. 
Table 2. Table Header for Pooled Analysis 
<Description> 
Placebo QW  
BIO89-100  
Total  
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Pooled 
Pooled 
2. A comparison between individual BIO89-100 dose group and the placebo group. 
Each BIO89-100 dose group will be compared with the placebo group. Due to the low 
number of subjects enrolled in the fibrate cohort, subjects randomized to BIO89-100 in 
fibrate cohort will be combined with the 27mg QW main cohort as the pooled 27mg QW 
group. The subjects randomized to placebo in the fibrate cohort will be combined with the 
placebo group. The table header is shown below (Table 3). The Total group will be 
presented as appropriate. 
Table 3. Table Header for Individual Dose Group Analysis 
<Description> 
Placebo QW  
Pooled 
BIO89-100  
9 mg QW 
 
BIO89-100  
18 mg QW 
 
BIO89-100  
27 mg QW  
Pooled 
BIO89-100  
36 mg Q2W 
Total 
For all other analyses, the results will be presented in a combined format as shown below 
(Table 3-1):  
Table 4-1. Table Header for All Other Analysis 
<Description> 
Placebo 
QW  
Pooled 
BIO89-100  
pooled 
 
BIO89-100  
9 mg QW 
 
BIO89-100  
18 mg QW 
BIO89-100  
27 mg QW  
Pooled 
BIO89-100  
36 mg 
Q2W 
Total 
5.6 PRE-TREATMENT MEDICATION AND PRE-TREATMENT PROCEDURES 
Any medication/procedure that starts and stops prior to 
. 
5.7 BASELINE MEDICATION 
Medications that have 
 within the duration of exposure will be considered 
baseline medications, i.e., medication taken prior to and/or on 
 and 
ongoing/stopped after 
. 
5.8 CONCOMITANT MEDICATION AND CONCOMITANT PROCEDURES 
Any medication/procedure that is taken any day on or after 
 is considered as 
concomitant medication/procedure. This includes medications/procedures that were started 
prior to administration of first dose and that were continued to be taken after 
. 
5.9 ADVERSE EVENT AND SERIOUS ADVERSE EVENT 
An AE is any untoward medical occurrence in a clinical study subject, temporally 
associated with the use of investigational product, whether or not considered as related to 
the investigational product. An AE can, therefore, be any unfavorable and unintended sign 
(including an abnormal laboratory finding), symptom, or disease (new or exacerbated) 
temporally associated with the use of investigational product. 
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A Serious Adverse Events (SAE) is defined as any untoward medical occurrence that, at 
any dose 1) results in death; 2) is life-threatening; 3) requires inpatient hospitalization or 
prolongation of existing hospitalization; 4) results in persistent disability/incapacity; 5) is 
a congenital anomaly/birth defect; 6) medically or scientifically judged as serious in other 
situations. 
5.10 TREATMENT EMERGENT ADVERSE EVENT 
TEAEs are AEs with onset date and time after or identical to the date and time of the first 
dose of the investigational product until the last dose + 28 days. 
6 ANALYSIS POPULATIONS 
The following analysis populations are defined for the main and fibrate expansion cohorts.  
Table 5. Analysis Population 
Population 
Definition 
Screened Analysis Set 
 
All subjects who signed informed consent and have undergone 
screening.  
Randomized Analysis Set 
All subjects in screened analysis set who are assigned a 
randomization number in the study. 
Full Analysis Set 
All randomized subjects who received at least 1 dose of 
investigational product, have a baseline, and at least 1 post-
baseline TG measurement not including EOS visit. 
Safety Analysis Set 
All subjects who received at least 1 dose of investigational 
product. 
Trough PK Analysis Set 
All subjects in the Safety Analysis Set, who have pre-dose and 
at least 1 post-dose trough PK measurements. 
Intensive PK Analysis Set  
All subjects in the Safety Analysis Set, who opted to participate 
in the Intensive PK scheme, and have at least 1 on-study PK 
measurement.  
MRI-PDFF Analysis Set 
All subjects in the Full Analysis Set who have baseline and a 
follow up MRI-PDFF assessments. 
All primary, secondary, and exploratory efficacy endpoints will be assessed for all subjects 
in the Full Analysis Set. All safety assessments will be displayed and analyzed for all 
subjects in the Safety Analysis Set. Intensive PK endpoints will be displayed and analyzed 
for all subjects with applicable data in the Intensive PK Analysis Set.  
6.1 
 HANDLING OF MISSING AND INCOMPLETE DATA 
6.1.1 Missing Dates 
For the missing or partially missing dates for the adverse event, prior/concomitant 
medication and procedures, the missing date will be imputed following the rules below. 
 
 
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6.1.2 Missing data for Efficacy Endpoints 
Missing data handling will be explored as sensitivity analysis if there is more than 10% 
missing data for the primary efficacy endpoint. The details are described in Section 7.5. 
 
 
 
 
6.2 ANALYSIS VISIT WINDOWS 
In general, all screening visits will be combined as Screening Analysis Visit. Scheduled 
visits from randomization (except for EOT visit for early terminated subjects) through 
/Week 8 will be used as analysis visit for descriptive analysis tables and listings.   
Unscheduled visits will be mapped according to the analysis visit window below.  If a 
scheduled visit is missing, the unscheduled visit that is closest to the target study day may 
be used for analysis purpose. 
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For early terminated subjects, EOT visits will be remapped to scheduled visit week using 
the actual study day and analysis window. EOS visits will be analyzed separately.  
Both nominal visits and analysis visits will be displayed in the listing. 
The visit schedules and window are shown below.  
Table 6. Analysis visit window 
Nominal Visit/Time 
point 
Analysis Visit Label 
Target Study 
Daya 
Analysis Windowa 
6.3 SOFTWARE 
All data processing, summarization and analyses will be performed using SAS Version 9.4 
or later of the SAS ® statistical software package. 
7 STATISTICAL METHODS OF ANALYSES 
7.1 GENERAL CONSIDERATIONS 
The statistical analysis for this study will be performed by combining the main cohort and 
the fibrate extension cohort. A single database lock will be implemented.  
All the tables and figures will be based on the pooled data from both cohorts, unless 
otherwise specified. One set of listings will be generated for the whole study.  
In general, summaries and analyses will be presented by dose/treatment group. Descriptive 
statistics will be presented for demographics and baseline characteristics, safety endpoints, 
and PK and PD parameters. Continuous variables will be summarized by number of 
subjects and mean, SD/standard error, median, minimum, and maximum values. Minimum 
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and maximum will be presented to the same number of decimal places as reported/collected. 
One additional decimal place will be presented for mean and median. Two additional 
decimal places will be presented for SD up to a max of three decimal places. 
Categorical variables will be summarized by number and percentage of subjects. 
Percentage will be presented to one decimal place. The number of missing values will be 
presented as a separate category with no percentage, but only if 1 or more subjects are 
missing data for the summary. Otherwise, all categories will be presented (even if no 
subjects are counted in the category). Counts of zero in any category will be presented 
without percentage.  
All statistical tests will be 2-sided and tested at a statistically significant level of 0.05 
without adjustment for multiplicity. Confidence intervals will be 2-sided 95%, unless stated 
otherwise. Two-sided p-values will be presented with four decimal places and values less 
than 0.0001 will be presented as <.0001. 
Both reported units (CN) and SI units will be presented in listings.  
Both CN and SI units will be used in lipid panel (TG and other lipids) tables and figures. 
For the rest, only CN units will be used for tables and figures, unless otherwise specified. 
Insulin will be presented with conventional unit of µIU/mL;  
The listings will only present non-imputed data. 
7.2 SUBJECT ACCOUNTABILITY 
The number of subjects screened and the number (percentage) of subjects who failed 
screening (i.e., subjects who consented to participate in the clinical study but were not 
subsequently randomized) and the reasons for screen failure will be summarized.  
In addition, the number of subjects who withdrew from the study and discontinued the 
investigational product will be summarized by discontinuation reason (including 
discontinuation due to COVID-19 pandemic). 
The information including demography and screen failure details will be listed. Subjects 
with dose interruptions affected by the COVID-19 pandemic will also be noted in the listing. 
7.3 PROTOCOL DEVIATIONS 
A full list of protocol violations and deviations will be compiled and reviewed by the 
clinical team to identify important violations/deviations before final database lock. For 
violations at study entry, subjects will be assessed against the inclusion and exclusion 
criteria of the protocol. For on-study deviations, compliance with the protocol will be 
examined using blinded review of the database with regard to prohibited therapies, and 
timing and availability of planned assessments. 
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7.4 INVESTIGATIONAL PRODUCT ADMINISTRATION 
7.4.1 Extent of Exposure and Treatment Compliance 
The length of exposure to investigational product (as BIO89-100 or placebo) will be 
calculated as the number of days from the first dose to the last dose of double-blind 
investigational product administration +7, regardless of if the subject missed one or more 
doses of investigational product.  
Total dose administered (mg) is calculated as number of injections received x dose (mg) of 
the investigational product contained in the injection.  
Overall study treatment compliance will be calculated at the end of treatment as  (total 
injections administered /total injections expected x 100%. Total injections expected is 
defined as the total number of injections protocol defined until the end of treatment.  
The length of exposure, number of actual drug injections and number of missed injections, 
total dose administered, and overall study treatment compliance will be summarized by 
treatment group using descriptive statistics for the safety population. The investigational 
product administration and compliance data, including reasons for poor compliance, will 
be listed for each subject. 
7.4.2 Medical History 
General medical history and background therapy will be summarized by treatment group, 
as well as overall, for FAS population and presented in a by-subject listing.  Where 
appropriate, terms will be coded using the Medical Dictionary for Regulatory Activities 
(MedDRA) Version 23.0 or higher.     
7.4.3 Demographics and Baseline Characteristics 
The following demographic and baseline characteristics will be summarized by treatment 
group, as well as overall, for Safety Analysis Set and for Full Analysis Set:  
• Age in years and by age group (21-<65, >=65-<75 and >=75)  
• Gender 
• Race  
• Ethnicity 
• Height (cm), weight (kg), weight group, body mass index (kg/m2), BMI group (< 
25, 25 -< 30, ≥ 30 kg/m2), 
 
  
• systolic and diastolic blood pressure (mmHg),  
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• Serum lipids, lipoproteins and apolipoproteins (TG [mg/dL], 
[mg/dL], VLDLC 
[mg/dL], VLDL-TG [mg/dL], LDL-C [mg/dL], HDL-C [mg/dL], non-HDL-C 
[mg/dL], 
 [mg/dL], ApoB [mg/dL],
 [mg/dL], 
 [mg/dL], 
) 
• hsCRP [g/dL]), Adiponectin, fasting plasma glucose, 
 
 
•  ALT (U/L), AST (U/L)  
• TG <750 mg/dL or ≥750 mg/dL (8.47 mmol/L)  
• Whether or not the subject is receiving background therapy (e.g., prescription fish 
oil and/or statins) 
• History of Type 2 Diabetes  
• History of hypertension 
• History of Non-alcoholic fatty liver disease (NAFLD)  
• History of Nonalcoholic steatohepatitis (NASH) 
• Estimated Glomerular Filtration Rate (eGFR) category (normal: 
≥90 mL/min/1.73m2; mild Renal Impairment: 60-89 mL/min/1.73m2; moderate 
Renal Impairment: 30-59 mL/min/1.73m2, and severe Renal Impairment: 15-29 
mL/min/1.73m2) 
• MRI-PDFF 
 
Data will be summarized using descriptive statistics for continuous variables and using 
counts and percentages for categorical variables by treatment group and overall.  
Hepatitis B surface antigen (HBsAg), hepatitis C virus (HCV), and serum pregnancy test 
(only for females of childbearing potential), follicle stimulating hormone (FSH) urine 
pregnancy test on Day 1 and thyroid-stimulating hormone (TSH) will be presented 
  
7.4.4 Prior, Baseline and Concomitant Medication/Procedure 
Medications will be coded using WHO Drug (March, 2020 or later). The frequency of use 
of prior medications and use of concomitant medications will be summarized by treatment 
group, as well as overall, for the safety population according to Anatomical Therapeutic 
Chemical (ATC) class and preferred term. Prior and concomitant medications will be listed 
for each subject.  
Prior and concomitant procedures will be listed. 
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7.5 EFFICACY ANALYSIS 
The objective for the primary efficacy analyses is to evaluate the effect of BIO89-100 on 
TG reduction after the 8-week treatment period.  The analyses consist of comparisons 
between the BIO89-100 groups and placebo as below: 
1. A pooled BIO89-100 group combining all dose groups will be compared with the 
placebo to evaluate the overall effectiveness of BIO89-100 
2. Individual BIO89-100 dose groups will be compared with the placebo group 
separately to evaluate the effectiveness of BIO89-100 at a specific dose 
The study is considered positive and meeting the primary objective if any of the 
comparisons above is statistically significant.  
All efficacy analysis will be performed using the FAS unless otherwise specified. Analyses 
comparing the placebo group and the pooled BIO89-100 group will be stratified if the 
sample size permits.  The analyses comparing the placebo and individual dose group will 
not be stratified due to anticipated small size. 
7.5.1 Analysis of Primary Efficacy endpoints 
The primary endpoint is the percent change in TG at Week 8 from baseline.  The TG value 
at week 8 is defined as the average of TG values at 
 
 In case of missing TG value at 
 the non-missing result will be used 
as the week 8 TG. The primary efficacy analysis will be performed using a mixed model 
with repeated measure (MMRM). The model includes the treatment group, baseline TG 
values, background therapy use, visit, visit and treatment interaction. The model will be 
fitted using all planned visits up to Week 8.  
If the mixed model assumption of the MMRM method is severely violated, the non-
parametric van Elteren test stratified by baseline TG level and background lipid therapy 
will be used to test the treatment difference using the pooled data.  The location shift 
estimate, and Hodges-Lehmann 2-tailed 95% confidence interval will be presented. For 
the comparison between the individual dose group and the placebo, the unstratified 
Wilcoxon rank-sum test will be used instead due to the low sample size.  
The primary efficacy analysis will be repeated in the subgroups (as noted in Section 4.7). 
 
 
 
 
   
 
 
 
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7.5.2 Analysis of Secondary Efficacy Endpoints 
Full analysis set will be used for all secondary efficacy analyses. The proportion of subjects 
with TG <500 mg/dL at Week 8 will be summarized by treatment group and analyzed using 
Cochran Mantel Haenszel (CMH) method stratified by the 2 stratification. Unstratified 
Chi-square test will be performed for comparisons between the placebo and the individual 
dose group or if the sample size within a stratum is too small (< 5 subjects). In addition, 
responder analysis at Week 8 will be performed to ensure the consistency of the results. 
The following threshold will be used to calculate: the proportion of subjects with TG 
normalized; with TG reduction by ≥30%, ≥40%, ≥50%, ≥60%, ≥70%, ≥80%, and ≥90% 
from baseline.  
Cumulative distribution function of both percentage changes from baseline and changes 
from baseline for TG at Week 8 by treatment group will be summarized descriptively and 
graphically. 
Percentage change in VLDL-C, LDL-C, non-HDL-C, HDL-C, VLDL-TG, ApoB, hsCRP, 
fasting plasma glucose, adiponectin, and body weight from baseline to Week 8 will be 
analyzed similarly as the primary endpoints. For parameters that only have one post-
baseline measurement, an ANCOVA model will be performed with treatment and 
background therapy as factors and baseline value as covariate. 
7.5.3 Analysis of Other Efficacy Endpoints 
Change, percent change in 
, ALT, body weight and MRI-PDFF at week 8 will be 
analyzed among those with TG reduction <30% vs. >=30%.  MMRM or ANCOVA models 
will be used accordingly. 
Tables of descriptive summary will be provided for the following. 
• 
 
 
 
 
 
 
 
 
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7.6 SAFETY ANALYSIS 
All safety analyses will be performed using the Safety Analysis Set.  Safety data presented 
by treatment group will be summarized on an ‘as treated’ basis.  Safety variables include 
treatment-emergent adverse events (TEAEs), concomitant medication use, physical 
examination, clinical laboratory parameters, vital signs, and ECG results. 
 for 
all safety analyses is defined as the date of the first dose of investigational product.  
7.6.1 Adverse Events 
Subject incidence of TEAEs will be tabulated by Medical Dictionary for Regulatory 
Activities (MedDRA, Version 23.0) system organ class, MedDRA preferred term, and 
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treatment groups. All TEAEs, all treatment-emergent related AEs, all TESAEs, and all 
treatment-emergent serious related AEs, the most severe CTCAE grade, TEAEs leading to 
IP discontinuation, and TESAEs leading to IP discontinuation will be summarized. Related 
is defined as “possibly related”, “probably related”, or “definitely related” to randomized 
investigational medicinal product. Subjects will be counted only once within a SOC and 
Preferred Term (PT), even if the subject experienced more than one TEAE within a specific 
SOC and PT, and PT only.  
All AEs, TESAEs, TEAEs leading to IP discontinuation, Fatal TEAEs and COVID-19 
related AEs will be listed. 
7.6.2 Clinical Laboratory Parameters 
The following laboratory assessments will be analyzed using the safety analysis set: 
Hematology: White Blood Cell (WBC) with differential (Total Neutrophils, Lymphocytes, 
Monocytes, Eosinophils, Basophils – absolute and %), Red blood cell (RBC) count, RBC 
Indices (Mean Corpuscular Volume [MCV], Mean Corpuscular Hemoglobin Concentration 
[MCH], Mean Corpuscular Hemoglobin Concentration [MCHC], Red Cell Distribution 
Width [RDW], Reticulocyte count) Hemoglobin, Hematocrit, Platelet count, Coagulation 
panel (including Prothrombin Time, International Normalized Ratio (INR), Activated 
Partial Thromboplastin Time [aPTT]) 
Lipid Assessment:
, ApoB, 
 non-HDL-C, LDL-C, 
 
, TG, VLDL-C, VLDL-TG. 
Clinical Chemistries: Alkaline Phosphatase (ALP), ALT, AST, Albumin, Bicarbonate, 
BUN (Blood urea nitrogen), Calcium, Creatinine, eGFR, Creatine Kinase (CK), Chloride, 
Fasting plasma glucose, Gamma-Glutamyl Transferase (GGT), Lactate dehydrogenase, 
Magnesium, Phosphorus, Potassium, Sodium, (Total bilirubin, Indirect/direct bilirubin), 
Total protein, Uric acid. For ALT, change and percent change from baseline at week 8 will 
be analyzed using MMRM method based on full analysis set. 
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 Principal 
Investigator (PI) or delegate. A listing will be presented for Urinalysis results. 
Other Study-Specific Laboratory Assessments: 
• BIO89-100 level (to be evaluated by bioanalytical laboratory)  
•  TSH level 
• Serum hCG pregnancy test, Urine hCG pregnancy test, FSH level 
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• Urine drug screen including amphetamines, barbiturates, cocaine metabolites, 
opiates, benzodiazepines, cannabinoids and phencyclidine 
• Virology - Human Immunodeficiency Virus (HIV) antibody, hepatitis B surface 
antigen (HBsAg, and HCV antibody). To determine current infection status, if 
HBsAg positive Hepatitis B Virus (HBV) DNA will be measured or if HCV 
antibody positive, HCV Ribonucleic Acid (RNA) will be measured. 
• 24-hour urine for cortisol, 
, 
, , total 
Insulin, hsCRP, 
, 
, Serum adiponectin, and 
 
. 
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. 
Laboratory results will be presented as received from the central laboratory (or kit provided 
by the central lab).  
Descriptive summaries by treatment groups will be produced for the laboratory tests.   
Shifts (low/normal/high) from the relevant baseline tables based on the normal ranges will 
be constructed.  The shifts will be performed from the baseline to each post-baseline visit, 
and to minimum and maximum post-baseline visit.  
7.6.3 Vital Signs 
The observed values and the change from baseline for vital signs (body temperature, pulse 
rate, RR, and supine BP) will be summarized by visit and treatment group. 
The number and percent 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 
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• Maximum decrease from baseline in DBP ≥20 mm Hg 
Clinically significant shifts for blood pressure from baseline to each post baseline visit will 
be summarized in the following criteria: 
Table 7. Blood Pressure Category 
Systolic (mm Hg) 
 
Diastolic (mm Hg) 
Blood Pressure Category 
>180 
Or 
>120 
Crisis 
≥140 
Or 
≥90 
Hypertension Stage 2 
130-139 
Or 
80-89 
Hypertension Stage 1 
120-129 
And 
<80 
Elevated 
<90 
Or 
<60 
Hypotension 
<120 
And 
<80 
Normal 
Clinically significant shifts for pulse rate from baseline to each post baseline visit will be 
summarized using the category as <50 bpm; 50-99 bpm, ≥100 bpm 
7.6.4 Physical Examinations 
Physical examination (abdominal, cardiovascular, neurological, respiratory, skin, and 
general, etc) results will be summarized by visits and by treatment group.  
Shift tables from baseline to end of the study will be summarized using the category as 
normal, abnormal but not clinically significant, and abnormal and clinically significant. 
7.6.5 Electrocardiograms  
 The following quantitative ECG measurements will be taken during the study: 
• heart rate (bpm); 
• PR interval (msec); 
• QRS interval (msec); 
• Fridericia corrected QT (QTcF) interval (msec)  
Descriptive summaries and changes from baseline will be presented by treatment group for 
the quantitative ECG measurements listed above and the qualitative overall ECG 
interpretation (categorized as normal; abnormal, insignificant; and abnormal, significant).  
Shift tables will present changes from baseline in ECG interpretation (categorized as 
normal; abnormal, not clinically significant; and abnormal, clinically significant) to each 
post-baseline visit.   
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7.7 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. 
7.8 PHARMACOKINETICS ENDPOINTS 
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.  
Individual and mean BIO89-100 serum concentrations for each nominal timepoints will be 
listed, summarized and plotted versus time. PK parameters will be listed for each subject 
in the analysis set and summary descriptive statistics will be presented.  
If data are suitable, PK parameters include: 
• 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) 
• Trough concentration 
Additional PK parameters may be calculated if also deemed appropriate. 
 
 
 
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8 LIST OF PLANNED TABLES, FIGURES 
The list of table, listing, and figure mockups are included in TLF Shell, as a separate 
document.  
 
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DocuSign Envelope ID: 16803ECF-6010-4C61-877C-946E5DEF0946


Protocol No: BIO89-100-221 V3.0 
 
Statistical Analysis Plan 
Version: 1.0 
 
Date: 03June2022 
Confidential 
Page 35 of 41 
 
9 APPENDICES  
Appendix 1 Schedule of Activities (SOA) 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Informed consent 
x 
Inclusion/exclusion 
x 
x 
x 
x 
Randomization 
x 
Demography, medical history 
x 
Physical examination 
x 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
xc 
x 
x 
Body weight 
x 
x 
x 
x 
x 
x 
x 
Heightd 
x 
BMI (derived) 
x 
x 
Vital signs (BPe, heart ratee, RR) 
and body temperature 
x 
x 
x 
x 
x 
x 
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DocuSign Envelope ID: 16803ECF-6010-4C61-877C-946E5DEF0946


Protocol No: BIO89-100-221 V3.0 
 
Statistical Analysis Plan 
Version: 1.0 
 
Date: 03June2022 
Confidential 
Page 36 of 41 
 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Hematology, blood chemistry and 
coagulation panel 
xf 
xf 
xf 
xf 
xf 
xf 
Urinalysis 
x 
x 
x 
x 
x 
Thyroid panel (TSH, free T4, and 
free T3) 
x 
x 
x 
x 
12-lead ECG 
x 
x 
x 
x 
x 
History of drug and/or alcohol 
abuse, urine drug test, and 
alcohol consumption test 
xg 
xg,h 
xh 
xh 
xh 
xh 
xh 
xh 
Serum lipids, lipoproteins and 
apolipoproteins (TG, 
, VLDL- 
C, VLDL-TG, LDL-C, HDL-C, 
non-HDL-C, 
RLP-C, 
 
ApoB, 
, 
) 
x 
xi 
x 
x 
xi 
xi 
x 
x 
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Protocol No: BIO89-100-221 V3.0 
 
Statistical Analysis Plan 
Version: 1.0 
 
Date: 03June2022 
Confidential 
Page 37 of 41 
 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Inflammatory biomarker 
(hsCRP) 
x 
x 
x 
x 
 and serum adiponectin, total 
x 
x 
x 
x 
Intensive PK (optional)k 
x 
x 
x 
x 
x 
x 
x 
x 
x 
Trough PK (for all subjects)k 
x 
x 
x 
x 
x 
HIV, HCV and HBV Serologyl 
x 
FSH if required to determine 
menopausal status 
x 
Serum pregnancy test (WOCBP) 
x 
x 
Urine pregnancy test (WOCBP) 
x 
x 
x 
x 
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DocuSign Envelope ID: 16803ECF-6010-4C61-877C-946E5DEF0946


Protocol No: BIO89-100-221 V3.0 
 
Statistical Analysis Plan 
Version: 1.0 
 
Date: 03June2022 
Confidential 
Page 38 of 41 
 
Study Period (Duration) 
Screening/ 
Lifestyle 
Stabilization 
(Min 
4 weeks to 
Max 
6 weeks) 
Screening/ 
TG 
Qualification 
(2 Week) 
Screening/ 
TG 
Sample 
Double-blind Treatment (8 Weeks) 
Follow-up 
(4 Weeks) 
Visits 
Study Dayb 
Jug dispensed for 24-hour urine 
collection 
for 
cortisol 
and 
creatininem 
x 
x 
Archived serum and plasma 
samplesn 
x 
x 
x 
Investigational product dosingp 
x 
x 
x 
x 
x 
x 
x 
x 
AE 
and 
SAEs 
including 
hypersensitivity-related AEsq 
x 
x 
x 
x 
x 
x 
Concomitant medication 
x 
x 
x 
x 
x 
x 
Review of lifestyle and alcohol 
user 
x 
x 
x 
x 
x 
x 
x 
x 
x 
MRI-PDFF s 
x 
x 
FGF21 
x 
Shaded columns indicate visits that can be conducted by home health providers. 
Abbreviations: AE = adverse event; 
; ALT = alanine aminotransferase; ALP = alkaline phosphatase;
; ApoB 
= apolipoprotein B100; 
 aPTT = activated partial thromboplastin time; AST = aspartate aminotransferase; 
 
 BP = blood pressure; BMI = body mass index; BUN = Blood urea nitrogen; CK = creatine kinase; CKD-EPI = chronic kidney disease epidemiology 
collaboration; 
; DNA = deoxyribonucleic acid; ECG = electrocardiogram; eGFR = estimated glomerular filtration 
rate; EOS = end of study; EOT = end of treatment; 
; FGF21 = fibroblast growth factor 21; FSH = follicle-stimulating hormone; GGT = 
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Protocol No: BIO89-100-221 V3.0 
 
Statistical Analysis Plan 
Version: 1.0 
 
Date: 03June2022 
Confidential 
Page 39 of 41 
 
gamma-glutamyl transferase; 
; HBsAg = Hepatitis B surface antigen; HCV = Hepatitis C Virus; HDL-C = high-density lipoprotein-
cholesterol; HHP = home health provider; HIV = Human Immunodeficiency Virus; hsCRP = high-sensitivity C-reactive protein; 
 
 INR = international normalized ratio; LDL-C = low-density lipoprotein-cholesterol; 
 non-HDL-C = non-high-density lipoprotein-
cholesterol; MCV = mean corpuscular volume; MCH = mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; MRI = Magnetic 
resonance imaging; MRI-PDFF = Magnetic Resonance Imaging – Whole Liver Proton Density Fat Fraction; 
; 
PK = pharmacokinetics; PT = prothrombin time; RNA = ribonucleic acid; RR = respiratory rate; RLP-C = remnant lipoprotein-cholesterol; SAE = serious adverse 
event; SoA = Schedule of Assessments; T3 = triiodothyronine; T4 = thyroxine; 
; TG = triglyceride; TSH = thyroid-stimulating hormone; V = 
Visit; VLDL-C = very low-density lipoprotein cholesterol; VLDL-TG = very low-density lipoprotein triglycerides; WBC = white blood cell; WOCBP = women of 
childbearing potential. 
Body posture during TG, BP, heart rate, RR and body temperature measurement needs to be the same (e.g., sitting, semi-erect, or supine) throughout the study with 
resting for at least 5 min prior to measurement at each study visit. 
Fasting blood samples should be taken after at least 12 hours of fasting. (It is recommended that subjects fast no more than 14 hours.) Fasting includes food and all 
beverages except for non-mineral water. 
Hematology: Hemoglobin, hematocrit, RBC count, reticulocyte count, MCV, MCH, MCHC, platelet count, WBC count, total neutrophils, eosinophils, monocytes, 
basophils, lymphocytes. 
Blood chemistry: BUN, creatinine (eGFR using CKD-EPI equation), fasting plasma glucose, calcium, sodium, potassium, chloride, total CO2 (bicarbonate), AST 
(SGOT), ALT (SGPT), ALP, GGT, total bilirubin, direct (conjugated) bilirubin, indirect (unconjugated) bilirubin, CK, uric acid, albumin, total protein. Coagulation 
panel: INR, aPTT, PT. 
Urinalysis: pH, glucose, protein, blood, ketones, nitrites, leukocyte esterase, urobilinogen, urine bilirubin, microscopy. 
a = If 
 is required, only assessment at that visit is TG measurement. 
b = 
 
 
c = Symptom-driven physical assessment for applicable reported AEs. Can be conducted on site during a scheduled or unscheduled visit or by a home health 
provider (HHP) if identified on the delegation log. d = Height at screening 
 only. 
e = Starting from randomization, pre-dose BP and pulse will be measured in duplicate. The first measurement will be initiated before dosing with sitting/semi-erect 
or supine position and resting for at least 5 minutes prior to measurements. BP and pulse are to be repeated at least 1 minute apart. Additional vital signs 
measurements may be done if clinically indicated. 
f = Coagulation panel will be done at 
 only. 
g = History of drug and/or alcohol abuse taken at screening 
 only. Urine drug test will be performed at 
 only. h= Alcohol breathalyzers will be used. 
Results >0.01% will be considered as positive. 
i = 
 
j = 
 
 
 
 
 
 
 
 
 
 
 
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Protocol No: BIO89-100-221 V3.0 
 
Statistical Analysis Plan 
Version: 1.0 
 
Date: 03June2022 
Confidential 
Page 40 of 41 
 
iii. 
 
 
k = Subjects will be included in either the Intensive PK scheme (optional) or Trough PK scheme (all subjects). Table 5 in the protocol details sampling times for 
the Intensive PK scheme. Intensive PK subjects will have additional visits on 
. For the Trough PK scheme, 
blood samples will be taken within 4 hours prior to dosing on specified dosing days or during EOT visit (
 for all subjects. 
Note: Where the Intensive PK schedule overlaps with the Trough PK schedule, only 1 PK measurement should be taken at those time points. 
l = Serology tests will include HBsAg, HCV, and HIV 1 and 2 antibodies. HBV and HCV determined by antibodies first and, if positive, by DNA/RNA. 
m = Ambulatory 24-hour urine collection for analysis of cortisol and creatinine to be initiated the day prior to 
 Subject will start collection 24 
hours before coming into the clinic and bring the sample to the visit. The 24-hour urine collection jug is dispensed at 
 so collection can be 
completed and brought to 
, respectively. The jug may be provided at the site or by an HHP. 
n = Archived for possible analysis of additional biomarkers. This is optional and by subject consent. 
o = 
 
 
 
p = Subjects randomized to BIO89-100 36 mg Q2W group will receive BIO89-100 at 
 and placebo at 
 to maintain the blind. 
Home health IP administration may occur on 
. 
q = When IP is scheduled to be administered by an HHP, a subject’s spontaneous reports of AE/SAE will be reported to the site staff who will follow-up and 
evaluate the need for an unscheduled visit for AE assessment. The sites/HHP may take non-personally identifying photographs of potential injection site 
reactions (optional). 
r = Study coordinators should reinforce maintenance of eating and exercise habits without weight loss or gain of 5% for the duration of the study, and alcohol 
consumption restrictions 48 hours prior to fasting blood work. 
s = After a consenting subject’s 
, the site should preemptively schedule the baseline MRI-PDFF to occur within the following projected visit windows to ensure 
an MRI appointment is available to be performed after the 
 TG result ≥350 mg/dL is known. For the main cohort, MRI-PDFF is optional and should occur 
prior to 
. For the fibrate expansion cohort, MRI-PDFF of ≥ 6.0% is required for enrollment and should be performed between 
 
. The follow-up MRI at 
 should only be performed if the baseline liver MRI-PDFF value is ≥ 6.0%. MRI-PDFF is mandatory in the 
fibrate expansion cohort. 
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Protocol No: BIO89-100-221 V3.0 
Statistical Analysis Plan 
Version: 1.0 
Date: 03June2022 
Confidential 
Page 41 of 41 
Appendix 2 SAS Sample Code 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
  
 
 
 
  
  
  
 
 
 
 
  
 
 
 
 
 
  
 
 
 
  
  
  
 
 
 
 
 
  
 
 
 
 
 
 
 
 
 
 
  
  
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
  
 
 
 
 
 
  
 
 
 
 
 
  
 
 
 
 
  
 
 
 
 
 
  
  
 
 
 
 
 
 
 
  
  
 
 
 
 
 
 
 
 
 
  
 
 
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