Clinical trial NCT05121441
Conditions: Obesity
Keywords: Obesity Anti-Obesity Agents Body Weight
Sponsor: Aardvark Therapeutics, Inc.
Start date: 2021-11-15; Completion date: 2022-11-09
Enrollment: 20 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
The purpose of this study is to evaluate safety and efficacy of twice-daily ARD-101 in obese subjects with a body mass index (BMI) of 30-45 kg/m2.

Detailed description:
This is a Phase 2, randomized, placebo-controlled study to investigate the effects of ARD-101, a small molecule targeting bitter taste receptors (TAS2Rs), in obese subjects with a body mass index (BMI) of 30-45 kg/m2. This study has a planned enrollment of 30 subjects and will be conducted in a single center in the United States.

The study will consist of a Screening Period (up to 28 days), a Treatment Period (28 days), and a Follow-up Period (End-of-Study Visit within 14 days after receiving the last dose of ARD-101). The screening procedures will be initiated upon completion of the informed consent process. Following completion of screening procedures and confirmation of eligibility, subjects will be enrolled to receive ARD-101 in an outpatient setting and will be instructed to visit the clinical center periodically for safety and efficacy assessments.

Arms: arm 1: Dose 200 mg of ARD-101, twice daily for 28 days arm 2: Placebo arm matching active arm ARD-101, 200 mg BID
Interventions: intervention 1: ARD-101 intervention 2: Placebo
Intervention descriptions: intervention 1: Twice daily, oral administration intervention 2: Twice daily, oral administration

Protocol documents:
Prot_SAP_000.pdf: 
Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 1 of 66 
09 May 2022 
 
Clinical Study Protocol 
A Phase 2, Placebo-Controlled, Randomized, Blinded Study to Evaluate the 
Safety, Tolerability and Preliminary Efficacy of ARD-101 in Adults with 
Obesity 
 
Sponsor: 
Aardvark Therapeutics 
4747 Executive Drive, Suite 1020 
San Diego, CA 92121 
Sponsor Study Number: 
AARD-201 
IND Number: 
142288 
IMP Name: 
ARD-101 
Development Phase: 
Phase 2 
Version (Date) of Protocol: 
Amendment 1 
Amendment 2 
Amendment 3 
Amendment 4 
Amendment 5 
1.0 (18 December 2020) 
2.0 (12 March 2021) 
3.0 (28 May 2021) 
4.0 (21 October 2021) 
5.0 (17 December 2021) 
6.0 (09 May 2022) 
 
 
 
 
 
This clinical study will be conducted in accordance with the International Council for Harmonization Tripartite 
Guideline for Good Clinical Practice (GCP) E6(R2), the protocol and with other applicable regulatory requirements. 
Confidentiality Statement 
This document contains confidential information of Aardvark Therapeutics. Do not copy or distribute without written 
permission from the Sponsor. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 2 of 66 
09 May 2022 
 
SIGNATURE PAGE 
Declaration of Sponsor  
Protocol Title: A Phase 2, Placebo-Controlled, Randomized, Blinded Study to Evaluate the 
Safety, Tolerability and Preliminary Efficacy of ARD-101 in Adults with Obesity 
This clinical study protocol was subjected to critical review. The information it contains is 
consistent with current knowledge of the risks and benefits of the investigational medicinal product 
(IMP), as well as with the moral, ethical and scientific principles governing clinical research as set 
out in the guidelines on GCP applicable to this clinical study. 
 
Sponsor Signatory 
 
 
Zhenhuan Zheng, PhD 
EVP of Research and Scientific Affairs 
Aardvark Therapeutics, Inc. 
 
Date 
 
May 12, 2022


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 3 of 66 
09 May 2022 
 
SIGNATURE PAGE 
Declaration of Sponsor 
Protocol Title: A Phase 2, Placebo-Controlled, Randomized, Blinded Study to Evaluate the 
Safety, Tolerability and Preliminary Efficacy of ARD-101 in Adults with Obesity 
This clinical study protocol was subjected to critical review. The information it contains is 
consistent with current knowledge of the risks and benefits of the investigational medicinal product 
(IMP), as well as with the moral, ethical and scientific principles governing clinical research as set 
out in the guidelines on GCP applicable to this clinical study. 
 
Sponsor Signatory 
 
 
Andreas Niethammer, MD, PhD 
Chief Medical Officer 
Aardvark Therapeutics, Inc. 
 
Date 
 
 
12MAY2022


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 4 of 66 
09 May 2022 
 
SIGNATURE PAGE 
Declaration of the Principal Investigator 
Protocol Title: A Phase 2, Placebo-Controlled, Randomized, Blinded Study to Evaluate the 
Safety, Tolerability and Preliminary Efficacy of ARD-101 in Adults with Obesity 
This clinical study protocol was subjected to critical review and has been released by the Sponsor. 
The information it contains is consistent with current risk and benefit evaluation of the IMP, as 
well as with the moral, ethical and scientific principles governing clinical research as set out in the 
guidelines on GCP applicable to this clinical study. 
 
Principal Investigator  
 
 
Principal Investigator 
 
 
Signature 
 
Date 
 
 
Site Number 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 5 of 66 
09 May 2022 
 
TABLE OF CONTENTS 
SIGNATURE PAGE ...................................................................................................................... 2 
SIGNATURE PAGE ...................................................................................................................... 3 
SIGNATURE PAGE ...................................................................................................................... 4 
TABLE OF CONTENTS ................................................................................................................ 5 
List of Tables ..................................................................................................................... 7 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS ................................................... 8 
1. 
PROTOCOL SUMMARY............................................................................................... 11 
1.1. 
Protocol Synopsis ............................................................................................... 11 
1.2. 
Schema ............................................................................................................... 16 
1.3. 
Schedule of Assessments, Time Points, and Window Allowance ..................... 17 
2. 
INTRODUCTION ........................................................................................................... 19 
2.1. 
Background......................................................................................................... 19 
2.2. 
Risk-benefit Assessment .................................................................................... 21 
2.2.1. 
Known Potential Risks.................................................................................... 21 
2.2.2. 
Known Potential Benefits ............................................................................... 21 
2.3. 
Summary of Findings from Non-clinical, in vitro Studies with Potential 
Clinical Relevance .............................................................................................. 22 
2.3.1. 
Non-clinical Pharmacology ............................................................................ 22 
2.3.2. 
Non-clinical Pharmacokinetics ....................................................................... 22 
2.3.3. 
Non-Clinical Safety Pharmacology and Toxicology ...................................... 24 
2.4. 
Summary of Findings from Previous Clinical Studies ....................................... 25 
2.4.1. 
Overview of Clinical Pharmacology ............................................................... 26 
2.4.2. 
Overview of Clinical Efficacy ........................................................................ 28 
2.4.3. 
Overview of Clinical Safety ........................................................................... 28 
2.4.4. 
Benefits and Risks Conclusions ...................................................................... 30 
3. 
STUDY ENDPOINTS ..................................................................................................... 31 
3.1. 
Primary Endpoint................................................................................................ 31 
3.2. 
Secondary Endpoints .......................................................................................... 31 
3.3. 
Exploratory Endpoints ........................................................................................ 31 
4. 
STUDY DESIGN ............................................................................................................ 33 
4.1. 
Overview ............................................................................................................ 33 
4.2. 
Justification for Dose .......................................................................................... 33 
4.3. 
Study Duration.................................................................................................... 33 
4.4. 
Study Completion ............................................................................................... 34 
4.5. 
Early Termination ............................................................................................... 34 
5. 
STUDY POPULATION .................................................................................................. 35 
5.1. 
Number of Subjects ............................................................................................ 35 
5.2. 
Inclusion Criteria ................................................................................................ 35 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 6 of 66 
09 May 2022 
 
5.3. 
Exclusion Criteria ............................................................................................... 36 
5.4. 
Restrictions ......................................................................................................... 38 
5.4.1. 
Dietary and Fluid Restrictions ........................................................................ 38 
5.4.2. 
Lifestyle Considerations ................................................................................. 38 
6. 
INVESTIGATIONAL MEDICINAL PRODUCT .......................................................... 39 
6.1. 
Premedication ..................................................................................................... 39 
6.2. 
Identity of the Investigational Medicinal Products ............................................ 39 
6.3. 
Supply, Packaging, Labeling and Storage .......................................................... 39 
6.4. 
Drug Accountability, Dispensing and Destruction ............................................. 39 
6.5. 
Dose Modification .............................................................................................. 40 
6.6. 
Stopping Rules.................................................................................................... 40 
6.7. 
Subject Identification.......................................................................................... 41 
6.7.1. 
Screening Numbers ......................................................................................... 41 
6.7.2. 
Enrollment Numbers ....................................................................................... 41 
6.8. 
Compliance ......................................................................................................... 41 
6.9. 
Special Precautions and Management of AEs .................................................... 41 
6.9.1. 
Eating disorders .............................................................................................. 41 
6.9.2. 
Allergic reactions ............................................................................................ 41 
6.9.3. 
Management of gastrointestinal disorders ...................................................... 42 
6.9.4. 
Embryo-fetal Toxicity..................................................................................... 42 
6.10. 
Concomitant Therapy ......................................................................................... 42 
6.10.1. 
Permitted Medications/Procedures ................................................................. 42 
7. 
DISCONTINUATION .................................................................................................... 43 
7.1. 
Subject Withdrawal and Replacement ................................................................ 43 
8. 
STUDY ASSESSMENTS AND PROCEDURES........................................................... 44 
8.1. 
Eligibility Screening ........................................................................................... 44 
8.2. 
Safety Variables.................................................................................................. 44 
8.2.1. 
Adverse Events ............................................................................................... 44 
8.2.2. 
Clinical Laboratory Assessments .................................................................... 49 
8.2.3. 
Vital Signs ...................................................................................................... 49 
8.2.4. 
Standard 12-lead Electrocardiograms ............................................................. 49 
8.2.5. 
Bioimpedance ................................................................................................. 49 
8.2.6. 
MRI PDFF and Full Body Composition Scan ................................................ 50 
8.2.7. 
Mixed Meal Tolerance Test (MMTT) ............................................................ 50 
8.2.8. 
Indirect Calorimetry ........................................................................................ 50 
8.2.9. 
Physical Examinations .................................................................................... 50 
8.2.10. 
Continuous Glucose Monitoring ..................................................................... 51 
8.2.11. 
Circulating Hormones ..................................................................................... 51 
8.2.12. 
Fecal Microbiome Analysis ............................................................................ 51 
8.2.13. 
Non-invasive Phenotypic Bitter Taste Test .................................................... 52 
9. 
STATISTICAL CONSIDERATIONS ............................................................................ 53 
9.1. 
Study Population ................................................................................................ 53 
9.1.1. 
Protocol Deviations ........................................................................................ 53 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 7 of 66 
09 May 2022 
 
9.2. 
General Considerations ...................................................................................... 53 
9.3. 
Subject Disposition ............................................................................................. 53 
9.4. 
Demographic and Anthropometric Information and Baseline Characteristics... 53 
9.5. 
Prior and Concomitant Medication and Drug Administration ........................... 54 
9.6. 
Exposure ............................................................................................................. 54 
9.7. 
Safety Analyses .................................................................................................. 54 
9.7.1. 
Adverse Events ............................................................................................... 54 
9.7.2. 
Clinical Laboratory Tests................................................................................ 55 
9.7.3. 
Vital Signs ...................................................................................................... 55 
9.7.4. 
Standard 12-lead Electrocardiogram .............................................................. 55 
9.7.5. 
Physical Examination ..................................................................................... 55 
9.8. 
Exploratory analysis ........................................................................................... 55 
9.9. 
Determination of Sample Size ............................................................................ 55 
10. 
ETHICAL, LEGAL AND ADMINISTRATIVE ASPECTS .......................................... 56 
10.1. 
Data Quality Assurance ...................................................................................... 56 
10.2. 
Data Collection and Access to Source Data/Documents .................................... 56 
10.3. 
Archiving Study Documents .............................................................................. 56 
10.4. 
Good Clinical Practice ........................................................................................ 57 
10.5. 
Informed Consent ............................................................................................... 57 
10.6. 
Insurance and Compensation for Injury ............................................................. 58 
10.7. 
Protocol Approval and Amendment(s) ............................................................... 58 
10.8. 
Confidentiality Data Protection .......................................................................... 58 
10.9. 
Publication Policy ............................................................................................... 59 
11. 
REFERENCE LIST ......................................................................................................... 60 
12. 
APPENDIX ..................................................................................................................... 61 
 
List of Tables 
Table 1 
Schedule of Assessments ......................................................................................... 17 
Table 2 
Caco-2 cell Monolayer Permeability of ARD-101, Warfarin, Ranitidine and 
Talinolol ................................................................................................................... 20 
Table 3 
The Pharmacokinetic Parameters of ARD-101 After Single Intravenous or 
Intragastric Administration to Cynomolgus Monkeys ............................................. 23 
Table 4 
 Geometric Mean (%CV) PK Parameters of ARD-101 Following Single Oral 
Dose Administration of ARD-101 ........................................................................... 26 
Table 5 
 Geometric mean (%CV) PK parameters of ARD-101 BID oral dose 
administration of ARD-101 for 14 days................................................................... 28 
Table 6 
Summary of Treatment-emergent Adverse Events - MAD Part .............................. 29 
Table 7 
Summary of Treatment-related Treatment-emergent Adverse Evens by System 
Organ Class and Preferred Term - MAD Part .......................................................... 30 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 8 of 66 
09 May 2022 
 
LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
Abbreviation 
Definition 
AE 
Adverse event 
ALT 
Alanine aminotransferase 
AST 
Aspartate transaminase 
AUC 
Area under the curve  
AUC0-τ 
Area under the plasma concentration versus time curve from time zero to 
time τ  
b.i.d. 
Twice a day 
BLQ 
Below the quantitation limit 
BMI 
Body mass index 
bpm 
Beats per minute 
CBC 
Complete blood count 
CCK 
Cholecystokinin 
CFR 
Code of Federal Regulations 
CGM 
Continuous glucose monitoring 
CHO 
Carbohydrate 
CL 
Systemic clearance  
Cmax 
Maximum observed concentration  
CNS 
Central nervous system 
CoEQ 
Control of Eating Questionnaire 
CRO 
Contract research organization 
CRP 
C-reactive protein 
CSR 
Clinical study report 
C-SSRS 
Columbia-Suicide Severity Rating Scale 
CYP 450 
Cytochrome P450  
ECG 
Electrocardiogram 
eGFR 
Estimated glomerular filtration rate 
EOS 
End of Study 
FIH 
First-in-Human 
F% 
Oral bioavailability 
FSH 
Follicle stimulating hormone 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 9 of 66 
09 May 2022 
 
Abbreviation 
Definition 
GCP 
Good Clinical Practice 
GI 
Gastrointestinal 
GIP 
Gastric inhibitory polypeptide 
GLP 
Good Laboratory Practice  
GLP-1 
Glucagon-like peptide-1 
HbA1c 
Glycated hemoglobin 
hERG 
Human ether-a-go-go-related gene 
HOMA-IR 
Homeostatic model assessment for insulin resistance 
HPBL 
Human peripheral blood lymphocytes 
ICF 
Informed consent form 
ICH 
International Council on Harmonization 
IC50 
Half maximal Inhibitory Concentration 
IEC 
Independent ethic committee  
i.g. 
Intragastric 
IgE 
Immunoglobulin E 
IMP 
Investigational medicinal product 
IL 
Interleukin 
IP 
Intraperitoneal 
IND 
Investigational new drug 
IRB 
Institutional review board 
i.v. 
Intravenous 
MATE 
Multidrug and toxic compound extrusion 
MDRI 
Multi-drug resistance-1 
MedDRA 
Medical Dictionary for Regulatory Activities 
MMTT 
Mixed Meal Tolerance Test 
MRI-PDFF 
Magnetic resonance imaging-derived proton density fat fraction 
NASH 
Nonalcoholic steatohepatitis  
NOAEL 
No observable adverse effect level  
OAT3 
Organic anion transporter 3 
OCT2 
Organic cation transporter 2 
OGTT 
Oral glucose tolerance test 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 10 of 66 
09 May 2022 
 
Abbreviation 
Definition 
OTC 
Over-the-counter 
PK 
Pharmacokinetics 
p.o. 
per. os (orally) 
PYY 
Peptide YY 
QTcB 
QT interval corrected for heart rate using Bazett’s correction 
QTcF 
QT interval corrected for heart rate using Fridericia’s correction 
SAE 
Serious adverse event 
SD 
Standard deviation 
SMC 
Safety Monitoring Committee 
SOC 
System organ class 
SOP 
Standard operating procedure 
SUSAR 
Suspected unexpected serious adverse reaction 
t1/2 
Terminal elimination half-life calculated as: ln2/λz  
tmax 
Time of the maximum observed concentration  
TEAEs 
Treatment-emergent adverse events 
TNF 
Tumor necrosis factor  
TSH 
Thyroid-stimulating hormone 
ULN 
Upper limit of normal 
US 
United States 
Vd 
Volume of distribution  
Vss 
Volume of distribution at steady state  
WHO 
World Health Organization 
WHO-DD 
WHO Drug Dictionary 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 11 of 66 
09 May 2022 
 
1. 
PROTOCOL SUMMARY 
1.1. 
Protocol Synopsis 
Protocol Title 
A Phase 2, Placebo-Controlled, Randomized, Blinded Study to Evaluate the 
Safety, Tolerability and Preliminary Efficacy of ARD-101 in Adults with 
Obesity 
Study Number 
AARD-201 
Development Phase 
Phase 2 
Sponsor 
Aardvark Therapeutics, Inc. 
Study Center 
The study will be conducted at a single center in the United States (US) 
Study Endpoints 
Primary Endpoint: 
• 
The percent total weight change at the end of treatment from baseline  
 
Secondary Endpoints: 
• 
Safety and tolerability of ARD-101 by assessment of the incidence of 
treatment-emergent adverse events (TEAE)  
• 
The change in blood lipid concentrations (total cholesterol, triglyceride, 
high density lipoprotein cholesterol, and low-density lipoprotein cholesterol) 
at the end of treatment from the baseline 
• 
The change in waist circumference at the end of treatment from the 
baseline 
• 
The change in hemoglobin A1c (HbA1c) at the end of treatment from the 
baseline 
 
Exploratory Endpoints: 
• 
Categorical weight loss. Proportion of subjects who lose < 5% and  ≥ 5% 
initial weight 
• 
Fasting circulating levels of glucagon-like peptide (GLP)-1 (total and 
active), cholecystokinin (CCK), peptide YY (PYY), amylin, glucose-
dependent insulinotropic polypeptide (GIP) (total and active), ghrelin, leptin, 
and adiponectin before the first dosing, and at 1h and 2h after the first dosing 
on day 1 
• 
Changes in response to mixed-meal tolerance test (MMTT), including: 
▪ 
Serum levels of glucose, insulin, and C-peptide prior to 
(negative timepoints) and post (positive timepoints) the Ensure 
meal given for MMTT at baseline (run-in visit) and on day 28.  
▪ 
Serum level of free fatty acids (FFA) prior to (negative 
timepoints) and post (positive timepoints) the Ensure meal given 
for MMTT at baseline (run-in visit) and on day 28  
▪ 
Circulating levels of GLP-1 (total and active), CCK, PYY, 
amylin, GIP (total and active), glucagon, and ghrelin prior to 
(negative timepoints) and post (positive timepoints) the Ensure 
meal given for MMTT at baseline (run-in visit) and on day 28  
▪ 
Circulating cytokines and inflammatory markers such as C-
reactive protein (CRP) at run-in visit and end of treatment (day 
28) performed during the MMTT 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 12 of 66 
09 May 2022 
 
▪ 
Circulating levels of bile acids, ferritin, leptin, and adiponectin 
at run-in visit and end of treatment (day 28) performed during 
the MMTT  
• 
Body composition as measured by bioelectrical impedance scale 
• 
Body composition and hepatic steatosis by magnetic resonance imaging-
derived proton density fat fraction (MRI-PDFF) + full body composition scan 
at pretreatment and end of treatment 
• 
The change in homeostatic model assessment for insulin resistance 
(HOMA-IR) at the end of treatment from the baseline   
• 
The change in fasting blood glucose at the end of treatment from the 
baseline   
• 
Hunger and control of eating as measured by Likert and visual analogue 
scales, respectively 
• 
Change in indirect calorimetry 
• 
The fecal samples will be collected  during the run-in visit and on day 28 
for microbiome analysis to evaluate potential effects of the study medication 
on fecal microbiome profile (optional for subjects) 
• 
Phenotypic bitter taste test (using commercially available test strips/kits) 
during the run-in visit of the study (optional for subjects) 
Study Design 
This is a Phase 2, randomized, placebo-controlled, single-blinded study of 
ARD-101 in obese subjects with Body Mass Index (BMI) of 30-45 kg/m2. 
 
A total of approximately 30 subjects will be randomized in a 2:1 ratio.  
 
A screening process will be initiated upon completion of the informed consent 
process. Subjects will be assessed for eligibility through screening tests 
conducted within 28 days prior to enrollment. Following completion of 
screening tests and confirmation of eligibility, subjects will be randomized to 
receive ARD-101 or placebo.  
 
All available safety, tolerability and preliminary efficacy data will be 
evaluated throughout study conduct. Pharmacodynamic data will be analyzed 
after study completion.  
 
Subjects will be receiving treatment in an outpatient setting and will be 
instructed to visit the clinical center periodically as scheduled in Table 1 for 
safety and efficacy assessments. The treatment may be discontinued due to 
intolerable toxicity, withdrawal of informed consent, discretion of principal 
investigator or end of the study.  
 
All subjects will be followed up for 14 days after the last dose of study drug. 
Test Product, Dose, and Mode of 
Administration 
ARD-101 200 mg BID administered orally for 28 days 
Inclusion Criteria 
Subjects must meet all the following criteria to be eligible for participation in 
this study: 
 
1. Male and female subjects, 18-75 years of age  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 13 of 66 
09 May 2022 
 
2. Willing and able to provide signed and dated informed consent prior to 
any study-related procedures and willing and able to comply with all study 
procedures 
3. BMI of 30-45 kg/m2 
4. Stable body weight by subject report (± 5%) in the previous 6 months 
prior to randomization 
5. No abnormal findings or abnormalities of clinical significance in vital 
signs, physical examination, clinical laboratory tests (complete blood 
count (CBC), urinalysis, blood biochemistry, coagulation, pregnancy test 
(females of child bearing potential), urine drug test, nicotine test, etc.), 
12-lead electrocardiogram (ECG) during the Screening Period. 
6. Serum creatinine, alkaline phosphatase, hepatic enzymes (aspartate 
aminotransferase, alanine aminotransferase) and total bilirubin (unless the 
subject has documented Gilbert syndrome) not exceeding 1.5-fold the 
upper laboratory norm and estimated glomerular filtration rate (eGFR) 
>30 mL/min 
7. Standard 12-lead ECG parameters after 10 minutes resting in supine 
position in the following ranges; 120 ms <PR <220 ms, QRS <120 ms, 
QTc ≤ 430 ms if male, ≤ 450 ms if female, and normal ECG tracing unless 
the Investigator considers an ECG abnormality within described limits to 
be not clinically relevant 
8. Stable or well controlled blood pressure per Investigator’s judgement 
during the Screening Period. Specifically: Vital signs after 10 minutes 
sitting in a chair (feet on floor, back supported): 
i. 95 mmHg <systolic blood pressure (SBP) <160 mmHg, 
ii. 45 mmHg <diastolic blood pressure (DBP) <100 mm Hg, 
iii. 40 bpm <heart rate (HR) <100 bpm 
9. Prediabetes- defined as a fasting blood glucose between 100-125 mg/dL 
OR an HbA1c between 5.7-6.5% at screening 
10. Type 2 diabetes- Defined as previous diagnosis by a healthcare 
professional OR a fasting blood glucose > 126 mg/dL OR HbA1c > 6.5% 
at screening.   
11. Patients with type 2 diabetes treated with metformin may be enrolled.  
However, patients with type 2 diabetes on any other therapy will be 
excluded.   
12. Female subjects must have negative pregnancy test and must not be 
lactating. For females able to bear children, a hormonal (i.e., oral, 
implantable, or injectable) and single barrier method (i.e., sponge), or a 
double-barrier method of birth control (i.e., condom with spermicide) or 
abstinence must be used/practiced throughout the study and for 90 days 
following last dose of study medication; for effective form of birth 
control. 
13. Females of non-childbearing potential, defined as surgically sterile (status 
post hysterectomy, bilateral oophorectomy, bilateral tubal ligation, 
bilateral salpingectomy, or bilateral tubal occlusion) or post-menopausal 
for at least 12 months (may be confirmed with a screening follicle 
stimulating hormone (FSH) level in the post-menopausal lab range), do 
not require contraception during the study. 
14. Males with female partners of childbearing potential must agree to a 
double-barrier method if they become sexually active during the study and 
for 90 days following the last dose of the study medication. Male subjects 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 14 of 66 
09 May 2022 
 
must not donate sperm for 90 days following their participation in the 
study. 
Exclusion Criteria 
Subjects who meet any of the following criteria will be excluded from study 
participation: 
 
1. History of significant drug hypersensitivity or anaphylaxis 
2. Prior bariatric or GI surgery (excluding cholecystectomy, 
hysterectomy or appendectomy) 
3. Participation in a weight loss program or clinical trial for weight 
loss within 30 days prior to randomization 
4. Diabetes treatment (unless metformin as outlined), or chronic oral 
steroids, or treatment with immune modulators, anti-obesity drugs, 
chronic opiate therapy, or antipsychotic medications.   
5. Received any experimental drugs or devices or have participated in 
a clinical study within 30 days prior to randomization 
6. Currently receiving any drug-based therapy for weight management  
7. Thyroid-stimulating hormone (TSH) level is outside of normal limit 
8. The presence of diseases with abnormal clinical manifestations that 
need to be excluded based on their possible contribution to weight 
loss or weight gain, including but not limited to nervous, 
cardiovascular, blood and lymphatic system, immune, renal, 
hepatic, gastrointestinal, respiratory, metabolic and skeletal diseases 
9. History of myocardial infarction, unstable angina, arterial 
revascularization, stroke, New York Heart Association Functional 
Class II-IV heart failure, or transient ischemic attack within 6 
months prior to Visit 1 
10. Any malignancy not considered cured (except focal, treated basal 
cell carcinoma and squamous cell carcinoma of the skin); a 
participant is considered cured if there has been no evidence of 
cancer recurrence in the previous 5 years 
11. History of major depressive disorder or history of other severe 
psychiatric disorders (e.g., schizophrenia or bipolar disorder) within 
the last 2 years. 
12. Major surgery within 3 months prior to randomization or planned 
surgery during the study 
13. Donated ≥200 mL of blood (blood components) or had massive 
blood loss, received blood transfusion or blood products within 3 
months prior to randomization 
14. Planned sperm/egg donation within 6 months post randomization 
15. Positive urine drug test for any illicit non-prescription substances 
16. History of consuming more than 14 units of alcoholic beverages per 
week or of alcoholism or drug/chemical/substance abuse within past 
2 years prior to enrollment (Note: one unit = 12 ounces of beer, 4 
ounces of wine or 1 ounce of spirits) 
17. Smoking any amount within 3 months prior to randomization 
18. Excessive consumption of tea, coffee, and/or caffeinated beverages 
(more than 8 cups, 250 mL for each cup) every day within 3 months 
prior to enrollment 
19. Symptomatic viral, bacterial (including upper respiratory infection), 
or fungal (non-cutaneous) infection within 1 week prior to 
randomization 
20. History of human immunodeficiency virus antibody or active 
hepatitis  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 15 of 66 
09 May 2022 
 
21. A history of psychiatric and psychological condition that, in the 
judgment of the investigator, may interfere with the planned 
treatment and follow-up, affect subject compliance or place the 
subject at high risk from treatment-related complications 
22. Poor venous access or inability to tolerate venipuncture 
23. Any condition or active drug treatment that the investigator or 
primary physician believes may not be appropriate for participating 
in the study 
Duration of Study 
The duration of participation for each subject will be approximately 84 days 
(12 weeks) 
• 
Screening Period: Up to 28 days (4 weeks) 
• 
Run-in Visit: Up to 14 days (2 weeks) 
• 
Treatment Period: 28 days on ARD-101 (4 weeks) 
• 
Follow-up Period: 14 days (2 weeks). The End of Study (EOS) Visit 
will occur within14 days after receiving the dose of ARD-101 
Pharmacodynamic Evaluation 
All subjects will be required to provide blood samples at baseline, at defined 
time points and at the end of treatment visit. These blood samples will be used 
to explore the parameters described in the endpoint setting. The Control of 
Eating Questionnaire (CoEQ) will be administered at the timepoints indicated 
in Table 1. 
Safety Evaluations 
Adverse events (AE) will be collected from the time of informed consent until 
14 days after the last administration of study drug. Safety laboratory 
assessments, including CBC, chemistries, coagulation tests, urinalysis, and 
thyroid function, along with ECGs, , and vital signs will be obtained. All 
concomitant medications will be recorded. All adverse events will be followed 
to resolution or stabilization. Only TEAE will be summarized. 
Efficacy Evaluations 
The efficacy assessments include changes from baseline in body weight, BMI, 
waist circumference, insulin resistance, body fat percentage, HbA1c, serum 
glucose, lipids, ambulatory blood pressure and defined hormone 
concentrations. 
Statistical Analysis 
All safety, and efficacy data will be summarized and presented overall for the 
study, using descriptive statistics. Mean, median, standard deviation, 
minimum, and maximum will be presented for continuous variables and   
frequencies and percentages for discrete variables. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 16 of 66 
09 May 2022 
 
1.2. 
Schema 
 
 
Assessed for eligibility 
Enrollment 
Randomization 
(N = 30) 
Follow up 
(14 days) 
Follow up 
(14 days) 
ARD-101 
(N = 20) 
200 mg BID X 28 days) 
Placebo 
(N = 10) 
(BID X 28 days) 
Analyzed 
Analyzed 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 17 of 66 
09 May 2022 
 
1.3. 
Schedule of Assessments, Time Points, and Window Allowance 
Details on procedures and timing of assessments are presented in Table 1. 
Table 1 
Schedule of Assessments 
Period 
Screening 
Run- in Visit 
Treatment Period  
(±3 days) 
Follow Up/End of 
Study 
Study Days 
-42 to -15 
14 days prior to day 1 
1a 
15 
28 
14 
Informed Consent 
X 
 
 
 
 
 
Inclusion/Exclusion Criteria 
X 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
Medical History 
X 
 
 
 
 
 
Randomization 
 
 
X 
 
 
 
Physical Examinationb 
X 
 
X 
X 
X 
X 
Vital Signs 
X 
X 
X 
X 
X 
X 
12-lead ECG 
X 
 
X 
 
X 
 
Hematology 
X 
 
Xc 
 
 
X 
Comprehensive Serum Chemistry 
X 
 
Xc 
X 
X 
X 
Urinalysis 
X 
 
 
 
 
X 
Urine Toxicology 
X 
 
 
 
 
 
Coagulation Parameters 
X 
 
Xc 
 
 
X 
Pregnancy Testd 
X 
 
X 
 
 
X 
TSH 
X 
 
 
 
 
 
Weight/BMI 
X 
X 
X 
X 
X 
X 
Bioimpedance 
 
 
X 
X 
X 
 
MRI PDFF + Full Body Composition Scan 
 
X 
 
 
X 
 
Height 
X 
 
 
 
 
 
Fasting Glucose 
 
X 
 
 
X 
 
HbA1c 
X 
 
 
 
X 
 
Waist Circumference 
 
 
X 
X 
X 
X 
Fasting Blood Lipids 
 
X 
 
 
X 
 
Fasting Circulating Hormonese 
 
 
X 
 
 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 18 of 66 
09 May 2022 
 
Period 
Screening 
Run- in Visit 
Treatment Period  
(±3 days) 
Follow Up/End of 
Study 
Study Days 
-42 to -15 
14 days prior to day 1 
1a 
15 
28 
14 
Study Drug Dispensation 
 
 
Xf 
 
 
 
Concomitant Medication 
X 
 
X 
X 
X 
 
Adverse Event Monitoring 
X 
 
X 
X 
X 
X 
Mixed Meal Tolerance Test (MMTT) 
 
X 
 
 
X 
 
CGM Placement 
 
X 
 
X 
 
 
CGM Removal and Download 
 
 
X 
 
X 
 
Indirect Calorimetry 
 
X 
 
 
X 
 
Fecal Sample Collectiong 
 
X 
 
 
X 
 
Phenotypic Bitter Taste Testh 
 
X 
 
 
 
 
Control of Eating Questionnaire (CoEQ) 
X 
 
X 
X 
X 
 
a. Predose 
b. Complete physical examination at screening, and brief physical examination for all follow-up visits 
c. This procedure is not needed if performed within 3 days 
d. For women of child-bearing potential only 
e. GLP-1 (total and active), CCK, PYY, amylin, GIP (total and active), ghrelin, leptin, and adiponectin in all subjects at 3 time points (before the first dosing 
and 1h as well as 2h after the first dosing) 
f. 
A single oral dose of ARD-101 will be administered to subject in the morning of day 1 
g. During run-in visit and on day 28. Optional for subjects 
h. Phenotypic bitter taste test is optional for sujects 
 
 
 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 19 of 66 
09 May 2022 
 
2. 
INTRODUCTION 
2.1. 
Background 
Obesity  
Obesity is a chronic disease with a multifactorial etiology including behavioral, environmental, 
sociocultural, physiological and genetic factors. obesity and its consequences are an increasing 
burden globally. Obese, defined as a BMI>30 kg/m2, is an established risk factor for a number of 
diseases such as hypertension, cardiovascular disease, cancers, diabetes type 2, and many others 
(Guh DP et al. 2009). Currently, there are an estimated 600 million obese people worldwide 
(Collaborators GBDO et al. 2017). BMI data shows that about 40% of American adults are obese 
(Hruby A and Hu FB 2015), while worldwide, about 3.4 million annual deaths are directly 
attributed to obesity (Collaborators GBDO et al. 2017).  
The treatment of obesity involves lifestyle changes, bariatric surgery and pharmacotherapy (Bray 
GA et al. 2016). Lifestyle change, including diet and exercise, requires considerable patient 
commitment, and has poor compliance. Surgery is expensive and invasive. Pharmacotherapy, 
therefore, is the primary option for the majority of patients. A variety of drugs are designed to 
prevent food absorption, decrease appetite, or increase metabolism. Compared to controls, most 
drugs cause weight loss of <4 kg with substantial severe adverse events (Rosa-Goncalves P and 
Majerowicz D 2019). Some of the adverse events have caused withdrawal of a number of drugs 
from the market in several countries (Rodgers RJ 2017). As a result, a significant portion of obese 
patients have no treatment options due to adverse events and limited weight loss. Taken together, 
obesity remains an unmet medical need and development of new compounds that are safe and 
effective is paramount.  
Denatonium 
Denatonium is one of the bitterest tastants among tens of thousands of natural or synthesized bitter 
compounds, with a bitterness detection threshold of 0.05 ppm. To date, one of the denatonium 
salts, denatonium benzoate has been approved as an additive to household products to prevent the 
ingestion of potentially harmful substances through taste aversion (Sibert JR and Frude N 1991). 
Importantly, the US Consumer Product Safety Commission has judged denatonium benzoate as 
generally safe. Like other bitter compounds, denatonium can strongly activate up to 25 G-protein 
coupled receptors expressed in a variety of tissues and organs including GI tract, pancreas, fat, 
trachea, and central nervous system. Bitter taste receptors are thought to be evolutionarily 
conserved for the detection of potentially toxic compounds.  Activation of bitter receptors in the 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 20 of 66 
09 May 2022 
 
mouth elicits a familiar aversive sensation, while activation of bitter receptors in the gut and 
chemoreceptor trigger zone located in the medulla leads to reduction of appetite and delayed 
gastric emptying as well as other adaptations. In a number of animal studies, denatonium benzoate 
has been shown to inhibit ongoing ingestive behavior, suppress food intake, and inhibit gastric 
emptying (Janssen S et al. 2011; Schier LA et al. 2011). Moreover, denatonium benzoate can 
stimulate the release of GLP-1 and CCK that are known to cause satiety and satiation (Chen MC 
et al. 2006; Kim KS et al. 2014). In human studies, the intragastric administration of denatonium 
benzoate after a liquid meal has been reported to inhibit relaxation of the proximal stomach. While 
in an oral nutrient tolerance test, denatonium benzoate was shown to increase satiation (Avau B et 
al. 2015). In a clinical trial conducted in healthy female volunteers, intragastric administration of 
denatonium benzoate is shown to decrease both antral motility and hunger ratings during the 
fasting state, probably via inhibiting motilin release. More importantly, no adverse reactions have 
been reported in all these human studies (Deloose E et al. 2017). Taken together, intragastric 
administration of denatonium salts is safe and can lead to weight loss by changing the appetite and 
eating behaviors and therefore, are promising agents for the treatment of obesity.  
Rationale of the Study 
ARD-101 is an encapsulated oral formulation of denatonium acetate salt that is designed to bypass 
the oral taste receptors when administered orally. Aardvark Therapeutics has demonstrated the 
efficacy and safety of ARD-101 in multiple preclinical animal models of metabolic disorders and 
weight management. The safety as well as pharmacokinetics have been confirmed in a human trial. 
In addition, ARD-101 is minimally absorbed into the circulation through the GI epithelium cells 
provided the very low cellular penetration rate as demonstrated by the Caco-2 cell monolayer 
permeability assay (Table 2). 
Table 2 
Caco-2 cell Monolayer Permeability of ARD-101, Warfarin, Ranitidine and 
Talinolol  
 
 
Compounds 
Papp (10-6 cm/s)  
Efflux ratio 
A→B 
B→A 
ARD-101 (10 µM) 
0.378 
4.65 
12.3 
Ranitidine 
0.355 
0.812 
2.29 
Warfarin 
34 
31 
0.913 
Talinolol 
0.337 
5.03 
14.9 
Talinolol + Verapamil 
0.737 
1.51 
2.05 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 21 of 66 
09 May 2022 
 
Taken together, ARD-101 has demonstrated promising preclinical activity in models of metabolic 
syndrome and a manageable toxicity profile in humans.  
For details of the preclinical results, please refer to the Investigator Brochure. 
2.2. 
Risk-benefit Assessment 
2.2.1. 
Known Potential Risks  
ARD-101 has shown a well-tolerated safety profile with no significant organ toxicities up to 200 
mg/kg/day in a GLP-compliant pivotal toxicology study with cynomolgus monkeys.   As with any 
new drug, rare or unexpected side effects could occur. The ARD-101 related toxicities may be 
mild to severe, including adverse hypersensitivities, or adverse reaction in GI system. 
Additionally, comprehensive non-clinical toxicology studies have been performed to establish the 
toxicity profile of ARD-101. The Cynomolgus monkey was identified as the pharmacologically 
relevant species for assessing human risk of ARD-101. ARD-101 was tolerated in a 
GLP-compliant, 28-day repeat-dose toxicology study in Cynomolgus monkey where systemic 
exposure was confirmed in all animals. 
No denatonium salt has been approved for treatment in subjects for any indication. Besides ARD-
101,denatonium benzoate is currently the only denatonium salt that has been tested in humans. 
The potential risks that may be associated with ARD-101 are considered to be similar to 
denatonium benzoate, which have been shown to be well-tolerated when administered p.o. (orally) 
with no reported adverse reaction (Deloose E et al. 2017). Taken together, denatonium salt is safe 
and well-tolerated when administered p.o. in humans. 
ARD-101 has been studied for its safety and pharmacokinetics in humans with the results in detail 
summarized below. In short, ARD-101 has been found to be safe at a dose level of up to 240 mg 
BID for 14 days and the PK has confirmed minimal systemic uptake and no accumulation after 11 
days. 
2.2.2. 
Known Potential Benefits 
Preclinically, ARD-101 was shown to reduce food and water consumption within 2 hours after 
dosing in SD rats. ARD-101 exhibited significant attenuation of weight gain (compared to 
control mice) in high fat diet-induced obese mice model after 56 days of treatment.  ARD-101 
also exhibited nominal attenuation of weight gain in ob/ob mice models.  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 22 of 66 
09 May 2022 
 
Further, previous studies, i.g. (intragastric) administration of denatonium helped in mitigation of 
chronic and acute inflammation in multiple preclinical models, characterized by improved 
pathology, downregulation of inflammatory cytokines (such as IL-6, TNF, IL-1β, etc.). Benefits 
were also observed with regards to cholesterol levels, HbA1c and other metabolic markers. 
In a 48-week study of diet-induced NASH in mice, dosing with ARD-101 largely abrogated 
development of liver fibrosis over control based on blinded histopathologic review. 
2.3. 
Summary of Findings from Non-clinical, in vitro Studies with Potential Clinical 
Relevance 
An in vitro secondary pharmacology study was performed to evaluate the binding, enzyme, and 
uptake activity of 10 µM ARD-101 and assess the specificity of ARD-101 activity. The study 
tested ARD-101 binding to 27 receptors (including subtypes of monoaminergic, cannabinoid, 
histaminic, nicotinic, opioid, and glucocorticoid receptors), 3 transporters, 8 ion channels, and 
6 enzymes. Results from 2 assays, α1A adrenergic receptor and hERG potassium ion channel 
assays, revealed an inhibition or stimulation higher than 50%, denoting a significant effect of 
ARD-101 in these two targets. The remaining assays did not show significant ARD-101 activity. 
2.3.1. 
Non-clinical Pharmacology 
Effect of ARD-101 to Support Proposed Indication 
The key data to support ARD-101 for the treatment of obesity and metabolic syndrome stems from 
preclinical experiments in multiple rodent models demonstrating benefit of oral dosing with ARD-
101 with regards to mitigation of weight gain as well as improvement in pathologies such as 
hypercholesterolemia, prediabetes and chronic inflammation. 
2.3.2. 
Non-clinical Pharmacokinetics 
The PK profile of ARD-101 was characterized in a panel of in vitro and in vivo studies, including 
plasma protein binding assays, permeability assays, metabolic stability and metabolite 
identification assessments, Cytochrome 450 (CYP450) phenotype assays, in vitro drug-drug 
interaction studies on CYP450s, and serum/plasma toxicokinetic studies in Cynomolgus monkeys.  
In the Caco-2 cell monolayer permeability assay, ARD-101 showed a low permeability through 
intestinal epithelia cell, the Papp value of ARD-101 in the apical to basolateral direction was 
0.378×10-6 cm/s. In addition, ARD-101 exhibited efflux effect with an efflux ratio of about 12.3, 
indicating ARD-101 is a substrate of certain efflux transporters.  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 23 of 66 
09 May 2022 
 
To investigate the PK of ARD-101 including systemic exposure, oral bioavailability, and PK 
linearity, a non-GLP PK study was conducted in Cynomolgus monkeys. Three male Cynomolgus 
monkeys were assigned to 3 dose groups and a single dose of ARD-101 was administrated p.o. at 
0.49, 1.62, or 4.85 mg/kg (denatonium base weight). After a 14-day washout period, an intravenous 
(i.v.) administration of ARD-101 was given to the same animal at the same dose level. Blood 
samples were collected from all animals on Day 1 and Day 15 at 0.25, 0.5, 1, 2, 4, 8, 24, 32, and 
48 h post-dose.  
Following a single i.v. dose of ARD-101 at 0.49, 1.62, and 4.85 mg/kg to Cynomolgus monkeys, 
the Cmax and AUC0-48h were 129, 803, 2270 ng/mL and 270, 1460, 4720 ng*h/mL, respectively. 
The values of CL and Vd were 2150, 1320, 1240 mL/h/kg and 32500, 25800, 17500 mL/kg, 
respectively. The elimination of ARD-101 in monkeys was slow, as indicated by the mean t1/2 of 
11.25 h. Within the dose range of 0.49-4.85 mg/kg, AUC increased higher than dose proportionally 
(Table 3).  
Following a single oral dose of ARD-101 at 0.49, 1.62, and 4.85 mg/kg, the AUC0-48h was 
insignificant and Cmax were 0, 1.98, 4.55 ng/mL, respectively, indicating an extremely low 
systemic exposure of ARD-101. The oral bioavailability (F%) in monkeys was estimated to be 0% 
at 0.49, 1.62, and 4.85 mg/kg, respectively. No clinical abnormal signs were noticed given very 
low absorption and negligible systemic exposure (Table 3).  
In summary, the bioavailability of ARD-101 in monkey is negligible, no systemic exposure 
reached after 4.85 mg/kg p.o. administration.  
Table 3 
The Pharmacokinetic Parameters of ARD-101 After Single Intravenous or 
Intragastric Administration to Cynomolgus Monkeys 
Route 
Dose 
(mg/kg) 
Cmax 
ng/mL 
AUC0-t 
ng*h/mL 
t1/2 
hours 
Tmax 
hours 
CL mL/min/kg 
VSS L/kg 
F% 
i.v. 
0.49 
129 
270 
105 
0.25 
2150 
32.5 
 
1.62 
803 
1460 
13.5 
0.25 
1320 
25.8 
 
4.85 
2270 
4720 
9.74 
0.25 
1240 
17.5 
 
p.o. 
0.49 
BLQ 
NA 
NA 
NA 
NA 
NA 
0 
1.62 
1.98 
NC 
NC 
2 
NA 
NA 
0 
4.85 
4.55 
NC 
NC 
1 
NA 
NA 
0 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 24 of 66 
09 May 2022 
 
Abbreviations: BLQ: below the quantitation limit; F: fraction (%) of an administered drug that reaches the systemic 
circulation [oral bioavailability]; i.v.: intravenous administration; p.o.: orally administration; NA: not applicable; 
NC: not calculable. 
Pharmacokinetic parameters after repeated oral doses were obtained from the toxicokinetic study 
following b.i.d. i.g. administration of 15, 30, or 100 mg/kg/dose of ARD-101 (denatonium base 
weight) to Cynomolgus monkeys for 28 days. After the first dose, the mean plasma AUC and Cmax 
values were similar between male and female animals, indicating a lack of sex-related differences 
in systemic exposure. There was a trend toward a greater than dose-proportional increase in Cmax 
and AUC0-24h. At Day 28, steady state mean AUC0-24h values were 26.3, 40.6, and 1210 ng*h/mL 
for female monkeys and 40.1, 48.0, and 1400 ng*h/mL for male monkeys at 15, 30, and 
100 mg/kg/doses, respectively. No accumulation of ARD-101 in monkeys was observed after 
4 weeks of b.i.d. administration.  
ARD-101 exhibited low binding (< 50%) to human and rat plasma proteins, and a relatively high 
binding to monkey plasma proteins (50% to 90%). An in vitro liver microsome stability study 
showed that ARD-101 was quickly metabolized by rat liver microsomes, moderately by monkey 
microsomes, and almost no metabolism in human liver microsomes. Cytochrome P450s (CYPs) 
were not the major enzymes involved in ARD-101 metabolism. An in vitro metabolite 
identification study showed that the N-dealkylated metabolite (M1, lidocaine) was only seen in the 
human S9 samples at trace levels, accounting for only 0.05% of total peak areas. The oxidation 
product M2 was the major metabolite, accounting for 18.88%, 5.82%, and 0.31% of total peak 
areas in rat, monkey and humans, respectively.  
A probe substrate-based inhibition study in human liver microsomes showed that ARD-101 was a 
moderate inhibitor of CYP2D6 with an IC50 value of 0.233 µM. The CYPs induction study in 
human hepatocytes showed that ARD-101 was not a potential inducer of CYP1A2, CYP2B6, and 
CYP3A4. An in vitro transporter interaction assay showed that ARD-101 is a substrate of MDRI, 
MATE1, MATE2-K, OAT3, and OCT2 transporters. ARD-101 is an in vitro inhibitor of the 
MATE1, MATE2-K, and OCT-2 transporters, with IC50 values of 21.07, 57.11, and 9.47 μM, 
respectively.  
2.3.3. 
Non-Clinical Safety Pharmacology and Toxicology 
The toxicology program was designed to support clinical studies in accordance with ICH Guidance 
M3(R2). A non-GLP single dose acute toxicology study (i.v. and p.o.) and a pivotal GLP compliant 
general toxicology study with a 28-day treatment (i.g.) and a 14-day recovery period have been 
conducted in Cynomolgus monkeys. In vitro studies including bacterial reverse mutation assay 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 25 of 66 
09 May 2022 
 
and mammalian cell chromosome aberration test have been conducted to evaluate genotoxicity of 
ARD-101.  
ARD-101 showed inhibition of hERG current with an IC50 of 99.2 µM (38143 ng/mL) in vitro. 
Cardiovascular toxicity was observed in a single Cynomolgus monkey after i.v. administration of 
ARD-101 at 4.85 mg/kg (in the single dose toxicology/PK study). The animal showed repetitive 
arrhythmia immediately following injection, the serum concentration was 2270 ng/mL at the first 
sampling time point (15 mins post-dose), and this adverse effect recovered by 8 hours post- dose. 
The tolerated dose for an i.v. route is 1.62 mg/kg with a systemic exposure of 1460 ng*h/mL and 
a peak serum concentration (Cmax) of 803 ng/mL.  
In the 28-day toxicity study in Cynomolgus monkeys, 100 mg/kg b.i.d. i.g. administration of 
ARD-101 (human equivalent dose is 64.5 mg/kg, b.i.d.) was well-tolerated in all animals, which 
is 37.33-fold of the proposed starting dose in a first-in-human (FIH) clinical study (40 mg b.i.d for 
a 70 kg adult). There was no mortality or moribundity found in ARD-101-treated animals. ARD-
101 showed no local toxicity in the GI organs (stomach, duodenum, ileum, jejunum, cecum, colon, 
and rectum) based on gross pathology and histological microscopic examinations. Only transient 
decreases in body weights in animals administered with 100 mg/kg dose were observed, which 
may be related to the pharmacological effect of ARD-101. The no-observed-adverse-effect level 
(NOAEL) was considered to be 100 mg/kg/dose (200 mg/kg/day) with a systemic exposure 
AUC0-24h of 1210 ng*mL/h and 1400 ng*mL/h for female and male monkeys, respectively. 
ARD-101 did not show mutagenic activity in the bacterial reverse mutation assay. The in vitro 
chromosomal aberration assay showed that ARD-101 was considered negative for inducing 
structural aberrations of human peripheral blood lymphocytes (HPBL) in the 3-h treatment with 
metabolic activation and the 22-hour treatment without activation. ARD-101 was positive for 
inducing structural aberrations of HPBL in the 3-hour treatment without metabolic activation. No 
precipitate, pH changes or cytotoxicity was observed at any dose level during the study. 
Further details can be found in the Investigator’s Brochure. 
2.4. 
Summary of Findings from Previous Clinical Studies 
A FIH clinical study with the study drug has been performed in healthy volunteers to interrogate 
safety and pharmacokinetics of multiple dose levels. 
The study consisted of 2 parts: A single ascending dose part (SAD) and a multiple ascending dose 
part (MAD), dosing subjects for 14 days BID. Each part tested three dose levels (40 mg, 100 mg 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 26 of 66 
09 May 2022 
 
and 240 mg) consecutively in a randomized, double-blinded manner. Forty-three subjects were 
dosed in total, 23 females and 20 males, of which 31 received ARD-101. 
The results are summarized below: 
2.4.1. 
Overview of Clinical Pharmacology 
2.4.1.1. 
Single Ascending Dose  
The SAD study evaluated PK parameters following administration of a single oral dose of ARD-
101 at 40 mg, 100 mg, or 240 mg in healthy adult subjects. Eligible subjects were 18-55 years of 
age, with a body mass index between 18-30 kg/m2 and a minimum weight of 50 kg. Subjects were 
orally dosed in 3 escalating cohorts of 40, 100, and 240 mg of ARD-101 or matching placebo 
capsules. For each dose cohort, 6 subjects received ARD-101 and 2 subjects received placebo. The 
study medication was administered to eligible subjects after a minimum of 8 h of fasting.  
Group PK analyses have been conducted for all dose cohorts in Part 1 of the study. PK parameters 
for all doses are presented in Table 4 . Based on the area under the curves (AUCs) and maximum 
serum concentration (Cmax) between-subject variability (Geometric CV%) was high, ranging from 
45.8 to 53.7%. CV% for t1/2 values increased with increases in ARD-101 dose level. 
Table 4 
 Geometric Mean (%CV) PK Parameters of ARD-101 Following Single Oral 
Dose Administration of ARD-101 
Parameter 
(Units) 
40 mg 
100 mg 
240 mg 
AUClast (h*ng/mL) 
4.81 (281.4) 
15.9 (61.1) 
53.1 (45.8) 
AUCinf (h*ng/mL) 
12.4 (106.8) 
17.5 (57.6) 
55.8 (45.2) 
Cmax (ng/mL) 
0.999 (259.4) 
3.04 (76.2) 
12.4 (53.7) 
tmaxa (h) 
1.33 (0.667 – 4.00) 
1.21 (0.667 – 2.00) 
1.08 (0.667 – 1.33) 
t1/2 (h) 
8.27 (14.4) 
11.1 (39.0) 
18.4 (39.0)b 
CL/F (L/h) 
3220 (106.8) 
5710 (57.6) 
4300 (45.2) 
Vz/F (L) 
38400 (114.0) 
91700 (30.1) 
114000 (56.9) 
a Median (min-max) presented; b Half-life estimates were calculated over a period of less than 2 half-lives for 3 
subjects in the 240 mg dose group.  
Following administration of a single 240 mg ARD-101 dose, quantifiable concentrations for all 
subjects were observed at 48 hours post dose, with 3 of 6 subjects having quantifiable 
concentrations at 72 hours post dose. There was a 2.4-fold increase in dose level from 100 to 240 
mg ARD-101. This resulted in increases in Cmax, AUClast, and AUCinf (4.08-, 3.34-, and 3.18-fold, 
respectively).  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 27 of 66 
09 May 2022 
 
The geometric mean and highest peak plasma concentrations (Cmax) following single oral 
administration at these doses were 0.999 and 9.59 ng/mL (40 mg), 3.04 and 10.4 ng/mL (100 mg), 
and 12.4 and 29.7 ng/mL (240 mg) respectively, which represented 0.12% and 1.20%, 0.380% and 
1.30%, and 1.55% and 3.71% of the PK stopping criteria (800 ng/mL) for this Phase I study. 
2.4.1.2. 
Multiple Ascending Dose  
The MAD study evaluated PK parameters following repeated administration to date of oral doses 
of ARD-101 at 40 mg, 100 mg and 240 mg BID, in healthy adult subjects. Eligible subjects were 
18-55 years of age, with a body mass index between 18-30 kg/m2 and a minimum weight of 50 kg. 
Subjects were orally dosed BID, for 14 days, or matching placebo capsules. 4 subjects received 
ARD-101 and 2 subjects received placebo in each dose level. From Day 1 to Day 14, the study 
medication was administered twice daily (every 12 h) with approximately 12 h between doses.  
PK results are listed in Table 5. 
The geometric mean (on Day 14) and highest individual systemic exposure (on Day 1), based on 
Cmax, represented 1.14% and 2.55% of PK stopping criteria, respectively, at the 240 mg BID dose 
level.  
Accumulation after BID dosing for 14 days was 1.20-fold for Cmax and 1.68-fold for AUCtau at 240 
mg BID. Steady state appears to have been reached by Day 11 at all tested dose levels.  
 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 28 of 66 
09 May 2022 
 
Table 5 
 Geometric mean (%CV) PK parameters of ARD-101 BID oral dose 
administration of ARD-101 for 14 days 
Parameter (Units) 
40 mg BID 
100 mg BID 
240 mg BID 
Day 1 
Day 14 
Day 1 
Day 14 
Day 1 
Day 14 
AUCtau (h*ng/mL) 
2.31 (187.9) 
5.51 (83.5) 
8.48 (63.0) 
16.4 (55.3) 
27.6 (80.3) 
46.3 (46.7) 
DAUCtau (h*ng/mL/mg) 
0.0577 
(187.9) 
0.138 
(83.5) 
0.0848 
(63.0) 
0.164 
(55.3) 
0.115 
(80.3) 
0.193 
(46.7) 
Cmax (ng/mL) 
0.584 (235.9) 
1.27 (141.3) 
1.85 (85.9) 
3.02 (86.1) 
7.64 (135.9) 
9.13 (59.6) 
DCmax (ng/mL/mg) 
0.0146 
(235.9) 
0.0317 
(141.3) 
0.0185 
(85.9) 
0.0302 
(86.1) 
0.0318 
(135.9) 
0.0380 
(59.6) 
tmaxa (h) 
1.50  
(0.50 – 4.00) 
1.75 
(1.08 – 4.00) 
1.50 
(1.08 – 2.00) 
1.08 
(0.50 – 4.00) 
1.75 
(1.15 – 3.00) 
1.08 
(0.50 – 2.00) 
Ctrough (ng/mL) 
NC 
0.147 (51.5) 
0.183 (69.3) 
0.481 (37.9) 
0.415 (76.0) 
1.21 (25.8) 
Cmin (ng/mL) 
NA 
0.119 (33.9) 
NA 
0.419 (25.1) 
NA 
1.21 (26.1) 
Cavg (ng/mL) 
NA 
0.459 (83.5) 
NA 
1.37 (55.3) 
NA 
3.86 (46.7) 
CLss/F (L/h) 
NA 
7260 (83.5) 
NA 
6090 (55.3) 
NA 
5180 (46.7) 
Racc, Cmax 
NA 
2.17 (495.5) 
NA 
1.66 (129.3) 
NA 
1.20 (50.1) 
Racc, AUCtau 
NA 
2.39 (69.2) 
NA 
2.02 (61.5) 
NA 
1.68 (26.9) 
Racc, Ctrough 
NA 
2.04 (21.5) 
NA 
2.89 (51.9) 
NA 
2.91 (45.9) 
a Median (min-max) presented; NC, not calculated 
 
2.4.2. 
Overview of Clinical Efficacy 
At the time of the submission of this IND, no clinical study has been conducted to investigate the 
efficacy of ARD-101. 
2.4.3. 
Overview of Clinical Safety 
Clinical safety is based on, three dose cohorts of the SAD part and three dose cohorts of the 
MAD part.  
In the SAD part of the study, ARD-101 was well-tolerated by subjects after single administration 
of the three assigned doses (40, 100, and 240 mg). No adverse effects (AEs) were reported for any 
subject in the three dosing cohorts. 
ARD-101 was also well-tolerated by subjects for 14-day repeated administration at 40 mg, 100 mg 
and 240 mg BID. Treatment-emergent AEs that were judged as treatment related by the 
investigator were restricted to Grade 1 or 2, and were experienced by a total of 6 subjects out of 
13 dosed with ARD-101 (46.2%) (Table 6). These consisted of gastrointestinal disorders (3 cases), 
nervous system disorders (3 cases), skin and subcutaneous disorders (2 cases), and cardiac 
disorders (1 case) (Table 7). Two cases of skin restricted hypersensitivity (urticaria) occurred at 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 29 of 66 
09 May 2022 
 
100 mg BID and 240 mg BID, respectively. The latter hypersensitivity was successfully addressed 
with premedication of oral antihistamine dosing as per protocol; while the subject experiencing 
Grade 1 hypersensitivity at the intermediate 100 mg BID dose was discontinued based on 
investigator’s decision and replaced. Gastrointestinal symptoms usually occurred, if so, early on 
and were self-limiting. One case of palpitations was reported lasting for about 3h. An ECG 
performed during that episode showed normal sinus rhythm at 83 bpm with no changes from 
baseline. 
Table 6 
Summary of Treatment-emergent Adverse Events - MAD Part 
 
 
 
 
Placebo (BID) 
(N = 6) 
nS (%) [nE] 
40 mg ARD-101 
(BID) 
(N = 4) 
nS (%) [nE] 
100 mg ARD-101 
(BID) 
(N = 5) 
nS (%) [nE] 
240 mg ARD-101 
(BID) 
(N = 4) 
nS (%) [nE] 
Overall 
(N = 19) 
nS (%) [nE] 
 
TEAEs 
Overall 
3 (50.0%) [5] 
1 (25.0%) [5] 
3 (60.0%) [5] 
3 (75.0%) [6] 
10 (52.6%) [21] 
Serious 
--- 
--- 
--- 
--- 
--- 
Leading to Discontinuation 
--- 
--- 
1 (20.0%) [1] 
--- 
1 (5.3%) [1] 
Leading to Death 
--- 
--- 
--- 
--- 
--- 
Severity 
--- 
--- 
--- 
--- 
--- 
Grade 1 
3 (50.0%) [5] 
1 (25.0%) [5] 
2 (40.0%) [3] 
2 (50.0%) [5] 
8 (42.1%) [18] 
Grade 2 
--- 
--- 
2 (40.0%) [2] 
1 (25.0%) [1] 
3 (15.8%) [3] 
Grade 3 
--- 
--- 
--- 
--- 
--- 
Grade 4 
--- 
--- 
--- 
--- 
--- 
 
Treatment-related TEAEs 
Overall 
1 (16.7%) [2] 
1 (25.0%) [4] 
2 (40.0%) [4] 
3 (75.0%) [6] 
7 (36.8%) [16] 
Serious 
--- 
--- 
--- 
--- 
--- 
Leading to Discontinuation 
--- 
--- 
1 (20.0%) [1] 
--- 
1 (5.3%) [1] 
Leading to Death 
--- 
--- 
--- 
--- 
--- 
Severity 
--- 
--- 
--- 
--- 
--- 
Grade 1 
1 (16.7%) [2] 
1 (25.0%) [4] 
1 (20.0%) [2] 
2 (50.0%) [5] 
5 (26.3%) [13] 
Grade 2 
--- 
--- 
2 (40.0%) [2] 
1 (25.0%) [1] 
3 (15.8%) [3] 
Grade 3 
--- 
--- 
--- 
--- 
--- 
Grade 4 
--- 
--- 
--- 
--- 
--- 
 
BID = twice daily; nE = number of adverse events; nS = number of subjects with an adverse event; N = number of subjects; % = percentage of su
bjects with an adverse event (nS/N×100) 
Severity grades: 1 = mild; 2 = moderate; 3 = severe; 4 = potentially life-threatening 
Adverse events were assigned severity grade using the Toxicity Grading Scale for Healthy Adult and Adolescent Volunteers Enrolled in Preventi
ve Vaccine Clinical Trials Version September 2007. 
A treatment-
emergent adverse event (TEAE) was defined as an adverse event that started during or after the first dose, or started prior to the first dose and incr
eased in severity after the first dose. 
A treatment-related TEAE was defined as a TEAE with a relationship of related to the study treatment, as determined by the investigator. 
Where a subject experienced multiple TEAEs with the same preferred term for the same treatment, this was counted as 1 TEAE for that treatment
under the maximum severity recorded. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 30 of 66 
09 May 2022 
 
Table 7 
Summary of Treatment-related Treatment-emergent Adverse Evens by 
System Organ Class and Preferred Term - MAD Part 
 
2.4.4. 
Benefits and Risks Conclusions 
According to our nonclinical studies, clinical data, and reported clinical studies on denatonium 
benzoate, ARD-101 has demonstrated a well-tolerated safety profile and potential clinical benefit 
in the treatment of obesity. This safety profile supports the use of the dose and dosing regimen to 
be used in the Phase 2 trial in overweight patients. 
 
 
System Organ Class 
 Preferred Term 
Placebo (BID) 
(N = 6) 
nS (%) 
40 mg ARD-101 
(BID) 
(N = 4) 
nS (%) 
100 mg ARD-101 
(BID) 
(N = 5) 
nS (%) 
240 mg ARD-101 
(BID) 
(N = 4) 
nS (%) 
Overall 
(N = 19) 
nS (%) 
Overall 
1 (16.7%) 
1 (25.0%) 
2 (40.0%) 
3 (75.0%) 
7 (36.8%) 
 
Gastrointestinal disorders 
1 (16.7%) 
1 (25.0%) 
1 (20.0%) 
1 (25.0%) 
4 (21.1%) 
 Diarrhoea 
--- 
--- 
1 (20.0%) 
1 (25.0%) 
2 (10.5%) 
 Dyspepsia 
--- 
1 (25.0%) 
--- 
1 (25.0%) 
2 (10.5%) 
 Nausea 
--- 
1 (25.0%) 
1 (20.0%) 
--- 
2 (10.5%) 
 Abdominal distension 
--- 
1 (25.0%) 
--- 
--- 
1 (5.3%) 
 Abdominal pain 
--- 
--- 
1 (20.0%) 
--- 
1 (5.3%) 
 Flatulence 
1 (16.7%) 
--- 
--- 
--- 
1 (5.3%) 
 
Nervous system disorders 
1 (16.7%) 
1 (25.0%) 
--- 
2 (50.0%) 
4 (21.1%) 
 Headache 
1 (16.7%) 
1 (25.0%) 
--- 
--- 
2 (10.5%) 
 Dizziness 
--- 
--- 
--- 
1 (25.0%) 
1 (5.3%) 
 Dysgeusia 
--- 
--- 
--- 
1 (25.0%) 
1 (5.3%) 
 
Skin and subcutaneous tissue 
disorders 
--- 
--- 
1 (20.0%) 
1 (25.0%) 
2 (10.5%) 
 Urticaria 
--- 
--- 
1 (20.0%) 
1 (25.0%) 
2 (10.5%) 
 
Cardiac disorders 
--- 
--- 
--- 
1 (25.0%) 
1 (5.3%) 
 Palpitations 
--- 
--- 
--- 
1 (25.0%) 
1 (5.3%) 
 
BID = twice daily; nS = number of subjects with an adverse event; N = number of subjects; % = percentage of subjects with an adverse event (nS/
N×100) 
Adverse events were coded using the Medical Dictionary for Regulatory Activities (MedDRA) Version 23.0. 
A treatment-
emergent adverse event (TEAE) was defined as an adverse event that started during or after the first dose, or started prior to the first dose and incr
eased in severity after the first dose. 
A treatment-related TEAE was defined as a TEAE with a relationship of related to the study treatment, as determined by the investigator. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 31 of 66 
09 May 2022 
 
3. 
STUDY ENDPOINTS 
3.1. 
Primary Endpoint 
• The percent total weight change at the end of treatment from baseline  
3.2. 
Secondary Endpoints 
• Safety and tolerability of ARD-101 by assessment of the incidence of TEAE  
• The change in blood lipid concentrations (total cholesterol, triglyceride, high density 
lipoprotein cholesterol, and low-density lipoprotein cholesterol) at the end of treatment 
from the baseline 
• The change in waist circumference at the end of treatment from the baseline 
• The change in HbA1c at the end of treatment from the baseline 
3.3. 
Exploratory Endpoints 
• Categorical weight loss. Proportion of subjects who lose < 5% and ≥ 5% of initial 
weight  
• Fasting circulating levels of GLP-1 (total and active), CCK, PYY, amylin, GIP (total 
and active), ghrelin, leptin, and adiponectin before the first dosing, and at 1h and 2h 
after the first dosing on day 1 
• Changes in response to MMTT, including: 
▪ Serum levels of glucose, insulin, and C-peptide prior to (negative timepoints) 
and post (positive timepoints) the Ensure meal given for MMTT at baseline 
(run-in visit) and on day 28  
▪ Serum level of FFA prior to (negative timepoints) and post (positive timepoints) 
the Ensure meal given for MMTT at baseline (run-in visit) and on day 28 
▪ Circulating level of GLP-1 (total and active), CCK, PYY, amylin, GIP (total 
and active), glucagon, and ghrelin prior to (negative timepoints) and post 
(positive timepoints) the Ensure meal given for MMTT at baseline (run-in visit) 
and on day 28 
▪ Circulating cytokines and inflammatory markers such as CRP at run in visit and 
end of treatment (day 28) performed during the MMTT 
▪ Circulating levels of bile acids, ferritin, leptin, and adiponectin at run in visit 
and end of treatment (day 28) performed during the MMTT  
• Body composition as measured by bioelectrical impedance scale 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 32 of 66 
09 May 2022 
 
• Body composition and hepatic steatosis by MRI PDFF + full body composition scan 
at pretreatment and end of treatment 
• The change in HOMA-IR at the end of treatment from the baseline   
• The change in fasting blood glucose at the end of treatment from the baseline   
• Hunger and control of eating as measured by Likert and visual analogue scales, 
respectively 
• Change in indirect calorimetry 
• The fecal samples will be collected during the run-in visit and on day 28 for 
microbiome analysis to evaluate potential effects of the study medication on fecal 
microbiome profile (optional for subjects) 
• Phenotypic bitter taste test (using commercially available test strips/kits) during the 
run-in visit of the study (optional for subjects) 
Already collected blood and fecal samples may also be stored and used for possible retrospective 
exploratory analysis which may include, but will not be limited to, understanding mechanisms of 
response to treatment (where response is defined broadly to include biomarker change, tolerability 
or safety) and identification of other mechanisms of therapy. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 33 of 66 
09 May 2022 
 
4. 
STUDY DESIGN 
4.1. 
Overview 
This is a Phase 2, randomized, placebo-controlled study of ARD-101 in subjects a BMI of 30-45 
kg/m2. 
This study has a planned enrollment of 30 subjects and will be conducted in a single center in the 
United States 
The study will consist of a Screening Period (up to 28 days), a Run-in Visite Period (up to 14 
days), a Treatment Period (28 days), and a Follow-up Period (EOS Visit within 14 days after 
receiving the last dose of ARD-101). The screening procedures will be initiated upon completion 
of the informed consent process. Following completion of screening procedures and confirmation 
of eligibility, subjects will be enrolled to receive ARD-101.  
ARD-101 will be administered orally under fasting conditions (approximately 60 minutes before 
breakfast and approximately 60 minutes before dinner) at 200 mg BID for 28 days. Subjects will 
receive treatment in an outpatient setting and will be instructed to visit the clinical center 
periodically as scheduled in Table 1 for safety and efficacy assessments. The treatment may be 
discontinued due to intolerable toxicity, withdrawal of informed consent, discretion of principal 
investigator or end of the study.  
4.2. 
Justification for Dose 
The dose of 200 mg BID was chosen based on the available nonclinical studies and clinical data. 
As described above, this dose has been found safe in the first-in-human trial and the 
pharmacokinetics support the prescribed dose level and duration. 
4.3. 
Study Duration 
The duration of participation for each subject will be up to approximately 84 days (12 weeks). 
• Screening Period: Up to 28 days (4 weeks) 
• Run-in Visit: Up to 14 days (2 weeks) 
• Treatment Period: 28 days on ARD-101 (4 weeks) 
• Follow-up Period: 14 days (2 weeks). The EOS Visit will occur within 14 days after 
receiving the dose of ARD-101 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 34 of 66 
09 May 2022 
 
4.4. 
Study Completion 
For the entire study, study completion is defined as the last visit of the last subject for any protocol 
related activity (last subject, last visit). For individual subjects, study completion is defined as the 
time of the subject’s last data collection. 
4.5. 
Early Termination 
If a subject withdraws prematurely after dosing, all data normally collected should be collected at 
the time of premature discontinuation. If deemed necessary by the Principal Investigator, the 
subject will be asked to return at the regularly scheduled EOS Visit. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 35 of 66 
09 May 2022 
 
5. 
STUDY POPULATION 
The study population will consist of male and female obese subjects with a BMI of 30-45 kg/m2. 
5.1. 
Number of Subjects 
A total of approximately 30 subjects will be enrolled in the clinical study, randomized 2:1 against 
placebo.  
If any assessments for inclusion or exclusion criteria are out of permissible ranges during the 
Screening Period, repeat assessments may be allowed for those subjects at Principal Investigator’s 
discretion. 
5.2. 
Inclusion Criteria 
Subjects must meet all the following criteria to be eligible for participation in this study: 
1. Male and female subjects, 18-75 years of age  
2. Willing and able to provide signed and dated informed consent prior to any study-related 
procedures and willing and able to comply with all study procedures 
3. BMI of 30-45 kg/m2 
4. Stable body weight by subject report (± 5%) in the previous 6 months prior to randomization 
5. No abnormal findings or abnormalities of clinical significance in vital signs, physical 
examination, clinical laboratory tests (CBC, urinalysis, blood biochemistry, coagulation, 
pregnancy test (females of childbearing potential), urine drug test, nicotine test, etc.), 12-lead 
ECG during the Screening Period. 
6. Serum creatinine, alkaline phosphatase, hepatic enzymes (aspartate aminotransferase, alanine 
aminotransferase) and total bilirubin (unless the subject has documented Gilbert syndrome) 
not exceeding 1.5-fold the upper laboratory norm and eGFR>30 mL/min 
7. Standard 12-lead ECG parameters after 10minutes resting in supine position in the following 
ranges; 120 ms <PR <220 ms, QRS <120 ms, QTc <=430 ms if male, <=450 ms if female and 
normal ECG tracing unless the Investigator considers an ECG abnormality within described 
limits to be not clinically relevant 
8. Stable or well controlled blood pressure per Investigator’s judgement during the Screening 
Period. Specifically: Vital signs after 10 minutes sitting in a chair (feet on floor, back 
supported): 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 36 of 66 
09 May 2022 
 
i. 95 mmHg < SBP <160 mmHg, 
ii. 45 mmHg < DBP <100 mm Hg, 
iii. 40 bpm < HR <100 bpm 
9. Prediabetes- defined as a fasting blood glucose between 100-125 mg/dL OR an HbA1c 
between 5.7-6.5% at screening 
10. Type 2 diabetes- Defined as previous diagnosis by a healthcare professional OR a fasting blood 
glucose > 126 mg/dL OR HbA1c > 6.5% at screening.   
11. Patients with type 2 diabetes treated with metformin may be enrolled.  However, patients with 
type 2 diabetes on any other therapy will be excluded.  
12. Female subjects must have negative pregnancy test and must not be lactating. For females able 
to bear children, a hormonal (i.e., oral, implantable, or injectable) and single barrier method 
(i.e., sponge), or a double-barrier method of birth control (i.e., condom with spermicide) or 
abstinence must be used/practiced throughout the study and for 90 days following last dose of 
study medication; for effective form of birth control. 
13. Females of non-childbearing potential, defined as surgically sterile (status post hysterectomy, 
bilateral oophorectomy, bilateral tubal ligation, bilateral salpingectomy, or bilateral tubal 
occlusion) or post-menopausal for at least 12 months (may be confirmed with a screening FSH 
level in the post-menopausal lab range), do not require contraception during the study. 
14. Males with female partners of childbearing potential must agree to a double-barrier method if 
they become sexually active during the study and for 90 days following the last dose of the 
study medication. Male subjects must not donate sperm for 90 days following their 
participation in the study. 
5.3. 
Exclusion Criteria 
Subjects who meet any of the following criteria will be excluded from study participation: 
1. History of significant drug hypersensitivity or anaphylaxis 
2. Prior bariatric or GI surgery (excluding cholecystectomy, hysterectomy or appendectomy) 
3. Participation in a weight loss program or clinical trial for weight loss within 30 days prior to 
randomization 
4. Diabetes treatment (unless metformin as outlined), or chronic oral steroids, or treatment with 
immune modulators, anti-obesity drugs, chronic opiate therapy, or antipsychotic medications.   


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 37 of 66 
09 May 2022 
 
5. Received any experimental drugs or devices or have participated in a clinical study within 30 
days prior to randomization 
6. Currently receiving any drug-based therapy for weight management  
7. TSH level is outside of normal limit 
8. The presence of diseases with abnormal clinical manifestations that need to be excluded based 
on their possible contribution to weight loss or weight gain, including but not limited to 
nervous, cardiovascular, blood and lymphatic system, immune, renal, hepatic, gastrointestinal, 
respiratory, metabolic and skeletal diseases 
9. History of myocardial infarction, unstable angina, arterial revascularization, stroke, New York 
Heart Association Functional Class II-IV heart failure, or transient ischemic attack within 6 
months prior to Visit 1 
10. Any malignancy not considered cured (except focal, treated basal cell carcinoma and squamous 
cell carcinoma of the skin); a participant is considered cured if there has been no evidence of 
cancer recurrence in the previous 5 years 
11. History of major depressive disorder or history of other severe psychiatric disorders (e.g., 
schizophrenia or bipolar disorder) within the last 2 years. 
12. Major surgery within 3 months prior to randomization or planned surgery during the study 
13. Donated ≥200 mL of blood (blood components) or had massive blood loss, received blood 
transfusion or blood products within 3 months prior to randomization 
14. Planned sperm/egg donation within 6 months post randomization 
15. Positive urine drug test for any illicit non-prescription substances 
16. History of consuming more than 14 units of alcoholic beverages per week or of alcoholism or 
drug/chemical/substance abuse within past 2 years prior to enrollment (Note: one unit = 12 
ounces of beer, 4 ounces of wine or 1 ounce of spirits) 
17. Smoking any amount within 3 months prior to randomization 
18. Excessive consumption of tea, coffee, and/or caffeinated beverages (more than 8 cups, 250 mL 
for each cup) every day within 3 months prior to enrollment 
19. Symptomatic viral, bacterial (including upper respiratory infection), or fungal (non-cutaneous) 
infection within 1 week prior to randomization 
20. History of human immunodeficiency virus antibody or active hepatitis  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 38 of 66 
09 May 2022 
 
21. A history of psychiatric and psychological condition that, in the judgment of the investigator, 
may interfere with the planned treatment and follow-up, affect subject compliance or place the 
subject at high risk from treatment-related complications 
22. Poor venous access or inability to tolerate venipuncture 
23. Any condition or active drug treatment that the investigator or primary physician believes may 
not be appropriate for participating in the study 
5.4. 
Restrictions 
5.4.1. 
Dietary and Fluid Restrictions 
Caffeine 
Subjects should not consume caffeine containing beverages exceeding 500 mg caffeine per day 
(5 cups of coffee) during the study. 
Alcohol 
Subjects should not consume more than 3 units (males) or 2 units (females) per day during the 
study (1 unit is equal to approximately ½ pint [284 mL] of beer, one small glass [125 mL] of wine, 
or one measure [25 mL] of spirits). 
5.4.2. 
Lifestyle Considerations 
Drugs of abuse 
Subjects must refrain from use of recreational drugs including THC containing products for the 
duration of the study. 
Nicotine 
No smoking or use of other nicotine-containing products (snuff, chewing tobacco, cigars, pipes or 
nicotine-replacement products such as nicotine chewing gum and nicotine plasters) will be allowed 
during the study. 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 39 of 66 
09 May 2022 
 
6. 
INVESTIGATIONAL MEDICINAL PRODUCT 
6.1. 
Premedication 
No pre-medication is required prior to administration of study medication. For subjects who 
experience Grade 2 acute allergic reactions and are able to continue on the treatment, 
premedication with an H1 antihistamine should be given. 
6.2. 
Identity of the Investigational Medicinal Products 
ARD-101 drug product is an encapsulated oral formulation of denatonium acetate monohydrate. 
The ARD-101 and placebo drug product will be supplied as white opaque body and white opaque 
cap hard gelatin capsules. The ARD-101 drug product contains 23.61 mg or 59.03 mg of 
denatonium acetate monohydrate (equivalent as 20 mg or 50 mg of denatonium base) together 
with povidone, sugar spheres, hypromellose, and talc as inactive ingredients. The inner content of 
the placebo has the same weight as the inner content of ARD-101 drug product, but only contains 
sugar spheres (no active ingredient). 
6.3. 
Supply, Packaging, Labeling and Storage 
Study drug and matching placebo will be supplied by Sponsor. The study medication will be 
packaged and labeled according to applicable local and regulatory requirements. 
All supplies of study medication must be stored in accordance with the manufacturer’s 
instructions. The ARD-101 will be stored at room temperature in a securely locked area, accessible 
to authorized persons only, until dispensation. 
Further details will be provided in the Pharmacy Manual. 
6.4. 
Drug Accountability, Dispensing and Destruction 
The Principal Investigator or designee is responsible for maintaining accurate accountability 
records of the study drug throughout the clinical study. Products returned will be stored under the 
same conditions as products not yet dispensed.  
All dispensing and accountability records will be available for Sponsor review. The Study Monitor 
will perform drug accountability at regular intervals.  
Study medication will be dispensed for each subject according to the protocol and Pharmacy 
Manual. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 40 of 66 
09 May 2022 
 
After receiving Sponsor approval in writing, all unused or partially used study medication will be 
returned to the Sponsor or designated third party or for preparing the study medication for 
destruction via incineration. 
6.5. 
Dose Modification  
The dose level may not be reduced. If any ≥ Grade 3 toxicities or any intolerable Grade 2 toxicity 
occurs, treatment should be discontinued permanently. Intolerable toxicity is defined as, in the 
opinion of Principal Investigator and Medical Monitor, any toxicity that poses a significant health 
risk to the subject. Treatment may continue on any Grade 1 toxic reactions or Grade 2 anorexia or 
Grade 2 weight loss. For any other Grade 2 toxicities, dosing may be interrupted for up to 3 days 
and may be resumed at the medical judgment of Investigator once the toxicity has resolved to 
Grade 0-1. A missed dose should not be made up. In the event of multiple toxicities, dosing 
management should be based on the worst toxicity observed. Subjects will be instructed to notify 
Investigators at the first occurrence of any adverse symptom. 
6.6. 
Stopping Rules 
At any time during the study, the subject must discontinue study drug treatment, if one of the 
following events occur:  
 
• An increase of AST and/or ALT (> 3x ULN) are observed in the presence of 
increased (> 2x ULN) alkaline phosphatase and/or total bilirubin.  
• A sustained AST or ALT increase, defined as three consecutive values of AST 
and/or ALT greater than 5x ULN within a period of 14 days.  
• A treatment-related Grade 3 or greater adverse event (other than AST or ALT 
increase as described above) is noted by the Investigator.  
• A relevant decrease of eGFR as per investigator 
  
If any of the above stopping rules are met the following action(s) should be taken:  
 
1. Stop dosing and convene a Safety Review Committee meeting;  
2. Review AE for evidence of relationship to treatment;   
3. May resume enrollment and continue the study if recommended by the Safety Review 
Committee.  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 41 of 66 
09 May 2022 
 
6.7. 
Subject Identification  
6.7.1. 
Screening Numbers 
All screened subjects are assigned a unique screening number. The screening numbers are numbers 
that identify subjects from time of screening until time of enrollment.  
6.7.2. 
Enrollment Numbers 
Eligible subjects will be assigned an enrollment number. The enrollment number will include 6-
digit site number and subject numbers starting with 101 as below.  
• Subject 100-101 
Once an enrollment number has been allocated to one subject, it may not be assigned to another 
subject.  
6.8. 
Compliance 
Dosing will be performed by trained, qualified personnel designated by the Principal Investigator.  
6.9. 
Special Precautions and Management of AEs 
6.9.1. 
Eating disorders 
ARD-101 may decrease appetite and subsequently change a subject’s eating behavior by delaying 
gastric emptying and inducing the glucagon-like peptide-1 and cholecystokinin secretion into the 
circulation that targets the central nervous system (CNS) to cause satiety/hunger sensation. 
Subjects who develop any eating disorders during study treatment should be permanently 
discontinued from study drug. 
6.9.2. 
Allergic reactions 
One case of allergic reaction has been reported in a female subject receiving denatonium benzoate, 
a compound in the same class as ARD-101 (Deloose E et al. 2017). Drug allergic reactions can be 
either acute or delayed. The acute allergic reactions that usually occur within 1 h after drug 
exposure are typically IgE mediated while those that usually occur 2 to 6 h after drug exposure 
have more complicated pathophysiological mechanisms. Subjects with a localized Grade 2 acute 
allergic reaction might continue on the treatment with premedication of H1 antihistamine. Subjects 
with recurrent Grade 2 acute allergic reactions with premedication, Grade 2 delayed allergic 
reactions, or Grade 3 to 4 acute allergic reactions should permanently discontinue the study drug.  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 42 of 66 
09 May 2022 
 
6.9.3. 
Management of gastrointestinal disorders 
Early recognition and management of potential GI disorders may mitigate more severe and 
subsequent toxicities.  
6.9.4. 
Embryo-fetal Toxicity 
The effect of ARD-101 on embryo-fetal development has not been tested. Thus, a highly effective 
form of contraception must be used, and enrolled subjects should receive instructions regarding 
this during the entire study from enrollment to 90 days after last dose administration. 
6.10. Concomitant Therapy 
Any medicinal product, prescribed or OTC, including vitamins, natural, and herbal remedies, taken 
by a subject other than the study medication, is considered concomitant medication. 
Any concomitant treatment will be given only if deemed strictly necessary by the Principal 
Investigator or designee. Use of concomitant medication will be recorded and reported. 
Particularly, as described in Section 2.3.2, ARD-101 is a substrate of the MDR1, MATE1, 
MATE2-K, OAT3 and OCT2 transporters. Coadministration of these transporter inhibitors have 
the potential to alter the pharmacokinetics of ARD-101. Therefore, before placing a subject on a 
specific medication/therapy, it is the responsibility of the investigator to check on potential drug-
drug interactions between that medication/therapy and ARD-101 based on the Drug Development 
and 
Drug 
Interactions: 
Table 
of 
Substrates, 
Inhibitors 
and 
Inducers 
(https://www.fda.gov/drugs/drug-interactions-labeling/drug-development-and-drug-interactions-
table-substrates-inhibitors-and-inducers), 
and 
Drug-Drug 
Interaction 
Database 
(https://www.fda.gov/drugs/drug-interactions-labeling/databases) of FDA. 
6.10.1. 
Permitted Medications/Procedures 
Subjects are permitted to use medications consistent with those permitted, including the use of 
contraceptive medications and hydrocortisone cream. All concomitant medications are subject to 
approval by the principal investigator. 
No concomitant use of approved obesity drugs or diabetes treatment (excluding metformin), 
MDR1, MATE1, MATE2-K, OAT3, or OCT2 inhibitors, or any drug deemed incompatible with 
trial participation as per investigator judgement.  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 43 of 66 
09 May 2022 
 
7. 
DISCONTINUATION 
7.1. 
Subject Withdrawal and Replacement 
Subject participation may be terminated prior to completing the study and the reason recorded as 
follows: 
  Adverse event 
  Protocol violation 
  Lost to follow-up 
  Subject withdrew consent at own request 
  Other 
A genuine effort must be made to determine the reason(s) why a subject fails to return for the 
necessary visits or is discontinued from the study. If the subject is unreachable by telephone, a 
registered letter, at the minimum, should be sent to the subject requesting him/her to contact the 
clinic. 
Withdrawn subjects may be replaced until the full cohort has completed the study. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 44 of 66 
09 May 2022 
 
8. 
STUDY ASSESSMENTS AND PROCEDURES 
For timing of assessments, refer to the Schedule of Assessments Table 1.  
8.1. 
Eligibility Screening 
All subjects must sign an informed consent form (ICF) prior to the conduct of any screening 
procedures. Screening assessments will occur within 28 days prior to the first dose of study 
medication. The subject’s relevant medical history and demographic data will be obtained. 
Retesting of abnormal screening values that lead to exclusion are allowed during the Screening 
Period (to reassess eligibility). The last result obtained prior to the first dose of study medication 
will be used to determine eligibility. 
8.2. 
Safety Variables 
8.2.1. 
Adverse Events 
Adverse event reporting will begin for each subject from the date the ICF is signed and will 
continue until the EOS Visit. 
8.2.1.1. 
Definitions 
8.2.1.1.1. 
Definition of Adverse Event 
Any untoward medical occurrence in a patient or clinical investigation subject administered a 
pharmaceutical product and which does not necessarily have a causal relationship with this 
treatment. An AE can therefore be any unfavorable and unintended sign (including an abnormal 
laboratory finding), symptom, or disease temporally associated with the use of a medicinal 
(investigational) product, whether or not related to the medicinal (investigational) product. 
Other untoward events occurring in the framework of a clinical study will be recorded as AEs, e.g. 
those occurring during treatment-free periods (including screening or post-treatment follow-up 
periods), in association with study-related procedures and assessments, or under placebo. For study 
medication, lack of efficacy may be an expected potential outcome and should not be reported as 
an AE unless the event is unusual in some way, e.g., greater in severity. 
Concomitant illnesses, which existed prior to entry into the clinical study, will not be considered 
AEs unless they worsen during the Treatment Period. Pre-existing conditions will be recorded as 
part of the subject’s medical history. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 45 of 66 
09 May 2022 
 
8.2.1.1.2. 
Definition of Serious Adverse Event 
An SAE is defined as any untoward medical occurrence that at any dose: 
• Results in death 
• Is life-threatening; this means that the subject was at risk of death at the time of the event; it 
does not mean that the event hypothetically might have caused death if it were more severe 
• Requires inpatient hospitalization or prolongation in existing hospitalization 
• Results in persistent or significant disability/incapacity or substantial disruption of the ability 
to conduct normal life functions 
• Is a congenital anomaly/birth defect, or 
• Is another important medical event (see below) 
Important medical events that do not result in death, are not life-threatening or do not require 
hospitalization may be considered SAEs when, based on appropriate medical judgment, they may 
jeopardize the subject and may require medical or surgical intervention to prevent one of the 
outcomes listed above. Examples of such medical events include allergic bronchospasm requiring 
intensive treatment in an emergency room or in a physician’s office, blood dyscrasias or seizures 
that do not result in in-patient hospitalization, and the development of drug dependency or drug 
abuse. 
A distinction should be drawn between serious and severe AEs. Severity is a measure of intensity 
whereas seriousness is defined by the criteria above. For example, a mild degree of gastrointestinal 
bleeding requiring an overnight hospitalization for monitoring purposes would be considered an 
SAE but is not necessarily severe. Similarly, an AE that is severe in intensity is not necessarily an 
SAE. For example, alopecia may be assessed as severe in intensity but would not be considered 
an SAE. 
Medical and scientific judgment should be exercised in deciding if an AE is serious and if 
expedited reporting is appropriate. 
8.2.1.2. 
Recording of Adverse Events 
Adverse events should be collected and recorded for each subject from the date the ICF is signed 
until the end of their participation in the study, i.e., the subject has discontinued or completed the 
study. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 46 of 66 
09 May 2022 
 
Adverse events may be volunteered spontaneously by the subject, or discovered by study staff 
during physical examinations or by asking an open, non-leading question such as ‘How have you 
been feeling since you were last asked?’ All AEs and any required remedial action will be 
recorded. The nature of AE, date (and time, if known) of AE onset, date (and time, if known) of 
AE outcome to date, severity and action taken of the AE will be documented together with the 
Principal Investigator’s assessment of the seriousness of the AE and causal relationship to study 
drug and/or study procedure. 
All AEs should be recorded individually in the subject’s own words (verbatim) unless, in the 
opinion of the Principal Investigator, the AEs constitute components of a recognized condition, 
disease or syndrome. In the latter case, the condition, disease or syndrome should be named rather 
than each individual symptom. The AEs will subsequently be coded using the Medical Dictionary 
for Regulatory Activities (MedDRA). Severity of AEs will be graded using the Toxicity Grading 
Scale for Healthy Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical Trials 
(September, 2007). 
8.2.1.3. 
Assessment of Adverse Events 
Each AE will be assessed by the Principal Investigator as per the categories discussed in the 
following sections. 
8.2.1.3.1. 
Intensity 
The Principal Investigator will assess all AEs for severity in accordance with the following 
standard ratings. 
• Mild (Grade 1): Ordinarily transient symptoms, does not influence performance of subject’s 
daily activities. Treatment is not ordinarily indicated. 
• Moderate (Grade 2): Marked symptoms, sufficient to make the subject uncomfortable. 
Moderate influence on performance of subject’s daily activities. Treatment may be necessary. 
• Severe (Grade 3): Symptoms cause considerable discomfort. Substantial influence on subject’s 
daily activities. May be unable to continue in the study and treatment may be necessary. 
• Potentially Life Threatening (Grade 4): Symptoms require emergency room (ER) visit or 
hospitalization. 
When changes in the intensity of an AE occur more frequently than once a day, the maximum 
intensity for the event should be noted for that day. Any change in severity of signs and symptoms 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 47 of 66 
09 May 2022 
 
over a number of days will be captured by recording a new AE, with the amended severity grade, 
and the date (and time, if known) of the change. 
8.2.1.3.2. 
Causality 
The Principal Investigator will assess the causality/relationship between the study medication and 
the AE. The Principal Investigator is responsible for assessing the relationship to study medication 
using clinical judgment and the following considerations: 
• No: Evidence exists that the AE has an etiology other than the study drug. For SAEs, an 
alternative causality must be provided (e.g., pre-existing condition, concomitant 
medication) 
• Yes: There is reasonable possibility that the event may have been caused by the study 
medication. 
8.2.1.4. 
Reporting of Serious Adverse Events 
The Principal Investigator will review each SAE and evaluate the intensity and the causal 
relationship of the event to study drug. All SAEs will be recorded from signing of the ICF until 
the EOS Visit. Serious AEs occurring after the EOS Visit and coming to the attention of the 
Principal Investigator must be reported only if there is (in the opinion of the Principal Investigator) 
reasonable causal relationship with the study drug. 
The Principal Investigator is responsible for providing notification to Sponsor of any SAE, whether 
deemed study medication-related or not, that a subject experiences during their participation in 
study within 24 h of becoming aware of the event. 
Initial reports of SAEs must be followed later with detailed descriptions, including clear 
photocopies of other documents as necessary (e.g., hospital reports, consultant reports, autopsy 
reports), with the subject’s personal identifiers removed.  
Information on SUSARs will be collected and reported to the regulatory authority and the IEC as 
required per guidelines. 
If the SUSAR is fatal or life-threatening, associated with the use of the study medication, and 
unexpected, the regulatory authority and the IEC will be notified per guidelines after being made 
aware of the case. Additional follow-up (cause of death, autopsy report, hospital report) 
information should be reported per guidelines. SUSARs which are not fatal and not life-threatening 
are to be reported per guidelines.  


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 48 of 66 
09 May 2022 
 
The Sponsor will notify the Investigators of relevant information about SUSARs that could 
adversely affect the safety of subjects in a timely fashion. Follow-up information may be submitted 
if necessary. The Sponsor will also provide annual safety reports for submission to the regulatory 
authority and the IEC responsible for the clinical study. These updates will include information on 
SUSARs and other relevant safety findings. 
8.2.1.5. 
Follow-up of Adverse Events 
All AEs experienced by a subject, irrespective of the suspected causality, will be monitored until 
the event has resolved, until any abnormal laboratory values have returned to baseline or stabilized 
at a level acceptable to the Principal Investigator and Medical Monitor, until there is a satisfactory 
explanation for the changes observed or until the subject is lost to follow-up. 
8.2.1.6. 
Pregnancy 
The Sponsor has a responsibility to monitor the outcome of all pregnancies reported during the 
clinical study. 
Pregnancy alone is not regarded as an AE unless there is a suspicion that the study drug may have 
interfered with the effectiveness of a contraceptive medication. Elective abortions without 
complications should not be regarded as AEs, unless they were therapeutic abortions (see below). 
Hospitalization for normal delivery of a healthy newborn should not be considered an SAE. 
Each pregnancy must be reported by the Principal Investigator to the Sponsor within 2 days after 
becoming aware of the pregnancy. The Principal Investigator must follow-up and document the 
course and the outcome of all pregnancies even if the subject was withdrawn from the clinical 
study or if the clinical study has finished. 
All outcomes of pregnancy must be reported by the Principal Investigator to the Sponsor on the 
pregnancy outcome report form within 2 days after he/she has gained knowledge of the normal 
delivery or elective abortion. 
Any SAE that occurs during pregnancy must be recorded on the SAE Report Form (e.g., maternal 
serious complications, therapeutic abortion, ectopic pregnancy, stillbirth, neonatal death, 
congenital anomaly, birth defect) and reported within 24 h in accordance with the procedure for 
reporting SAEs. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 49 of 66 
09 May 2022 
 
8.2.2. 
Clinical Laboratory Assessments 
Samples for clinical laboratory assessments will be collected at the time points detailed in the 
Schedule of Assessments (Table 1). Samples will be collected in appropriate tubes and handled 
according to standard procedures of the applicable laboratory. 
Any value outside the normal range will be flagged for the attention of the Principal Investigator 
or designee at the site. The Principal Investigator or designee will indicate whether the value is of 
clinical significance. Additional testing during the study may be done if medically indicated. If a 
clinically significant abnormality is found in the samples taken after dosing, during the study, 
and/or at the EOS Visit, it should be recorded as an AE and the subject will be followed until the 
test(s) has (have) normalized or stabilized, at the discretion of the Principal Investigator. 
8.2.3. 
Vital Signs 
Vital signs will be assessed at the time points detailed in the Schedule of Assessments Table 1. 
The following vital signs will be measured: 
• Blood pressure (systolic and diastolic [mmHg]) 
• Pulse (bpm) 
• Body temperature (°C) 
• Respiratory rate (breaths per minute) 
8.2.4. 
Standard 12-lead Electrocardiograms 
Standard safety 12-lead ECGs will be performed at the time points detailed in the Schedule of 
Assessments (Table 1). 
The ECG will include all 12 standard leads.  
All ECGs must be evaluated by a qualified physician for the presence of abnormalities. If a 
clinically significant abnormality is found, it should be recorded as an AE and the subject will be 
followed until the test(s) has (have) normalized or stabilized, at the discretion of the Principal 
Investigator. 
8.2.5. 
Bioimpedance 
Body composition will be assessed via scale measuring bioimpedance at the time points detailed 
in the Schedule of Assessments (Table 1). 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 50 of 66 
09 May 2022 
 
8.2.6. 
MRI PDFF and Full Body Composition Scan 
In order to better assess overall body composition including quantification of lean body mass, liver 
fat content and other related parameters, MRI scan will be performed at the time points detailed at 
the time points detailed in the Schedule of Assessments (Table 1). 
8.2.7. 
Mixed Meal Tolerance Test (MMTT) 
The MMTT permits measurement of intestinal (incretin secretion), β-cell (insulin secretion), 
hepatic (insulin extraction), muscle, and adipose tissue responses to ingesting calories. The mixed 
meal, containing glucose, fat and protein (2x Ensure 8 oz. for a total of 16 oz., 66 grams 
carbohydrate (CHO), 440 calories), is more physiologic than the glucose only challenge in an oral 
glucose tolerance test (OGTT). The MMTT will be performed at the time points detailed in the 
Schedule of Assessments (Table 1) and as described in the Appendix.  
8.2.8. 
Indirect Calorimetry 
Continuous indirect calorimetry using a commercial instrument (Vmax Encore VS 29N, 
CareFusion, Yorba Linda, CA, USA) will be performed during basal and postprandial periods on 
the days when the MMTT are performed.   
At -80 and 195 minutes during the MMTT, the canopy will be placed over the subject’s head to 
collect data from 10 continuous minutes at steady-state (no longer than 30 minutes) during each 
interval for determination of the substrate oxidation rates at basal condition, and during the MMTT 
ass outlined in the Schedule of Assessments (Table 1). 
8.2.9. 
Physical Examinations 
Physical examinations will be performed at the time points detailed in the Schedule of Assessments 
(Table 1). 
Full physical examination: 
An assessment of general appearance and a review of systems (dermatologic, head, eyes, ears, 
nose, mouth/throat/neck, thyroid, lymph nodes, respiratory, cardiovascular, gastrointestinal, 
extremities, musculoskeletal, neurologic and psychiatric systems). 
Brief physical examination: 
An assessment of the general appearance, skin, cardiovascular system, respiratory system and 
abdomen. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 51 of 66 
09 May 2022 
 
The brief physical examination may be extended to a full physical examination if considered 
necessary by the Principal Investigator. Other evaluations may be performed as deemed necessary 
by the Principal Investigator.  
8.2.10. 
Continuous Glucose Monitoring 
Assessments will be performed at the time points detailed in the Schedule of Assessments (Table 
1). 
Glucose levels will be monitored using the Continuous Glucose Monitoring (CGM) device. The 
CGM device will be used to track blood sugar levels day and night, and it will collect readings 
automatically every 5 minutes. The CGM sensor is a round disk with the circumference equal to 
that of a quarter, with the thickness of 2 quarters. It has a small filament that is inserted 
subcutaneously via a small needle that is then retracted and removed, leaving just the filament.  
The CGM sensor adheres to the skin leaving the filament in place for 14 days and stores the 
recorded glucose readings until downloaded by a study team member.   
During the Run-in Period, the CGM sensor will be applied to the subject and they will be required 
to wear it for a minimum duration of two weeks prior to Treatment Period Day 1. Data from the 
CGM will be downloaded and a report will be generated using the software provided by the device. 
The report outlines typical measures of glycemic control: mean glucose, standard deviation, time 
in range and daily patterns. In addition, a raw data file is available with each time point of 
collection and glucose reading.”   
8.2.11. 
Circulating Hormones 
To investigate the effect of the study medication on circulating gut hormone levels, all subjects of 
the study will undergo a fasting blood draw before the first dose, and 1h and 2h after the first dose 
on day 1 for measuring fasting circulating levels of the following gut hormnones: GLP-1 (total and 
active), CCK, PYY, amylin, GIP (total and active), ghrelin, leptin, and adiponectin (Table 1). 
8.2.12. 
Fecal Microbiome Analysis 
All subjects of the study will optionally provide their fecal samples during the run-in visit and on 
day 28 for microbiome analysis to evaluate potential effects of the study medication on fecal 
microbiome profile (Table 1). 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 52 of 66 
09 May 2022 
 
8.2.13. 
Non-invasive Phenotypic Bitter Taste Test 
All subjects of the study will optionally undergo a non-invasive phenotypic bitter taste test using 
commercialized phenylthiocarbamide (PTC) and 6-n-propylthiouracil (PROP) taste strips, and a 
commercialized Allegro Bitrex® Fit Test Kit during the run-in visit of the study (Table 1). This 
data can be used to retroactively assess a correlation of bitter taste sensitivity of subjects with the 
efficacy of the study medication. 
 
 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 53 of 66 
09 May 2022 
 
9. 
STATISTICAL CONSIDERATIONS 
9.1. 
Study Population 
9.1.1. 
Protocol Deviations 
Important protocol deviations will be listed by subject. 
9.2. 
General Considerations 
Continuous data per treatment arm will be summarized using descriptive statistics (number, mean, 
standard deviation [SD], minimum, median and maximum). Categorical data will be summarized 
using frequency tables (number and percentage). 
9.3. 
Subject Disposition 
Subjects excluded from the safety analysis sets will be listed including the reason for exclusion. 
Subject disposition will be summarized and will include the following information: Number of 
subjects dosed, number and percentage of subjects completing the study and the number and 
percentage of subjects who were withdrawn (including reasons for withdrawal). Disposition data 
will be presented based on all. 
Subject discontinuations will be listed including the date of study exit, duration of treatment and 
reason for discontinuation. A listing of informed consent response will also be presented. 
9.4. 
Demographic and Anthropometric Information and Baseline Characteristics 
Demographic and anthropometric variables (age, sex, ethnicity, race, height, body weight and 
BMI) will be listed by subject. Demographic characteristics (age, sex, ethnicity and race) and 
anthropometric characteristics (height, body weight, and BMI) will be summarized for all subjects 
in the safety analysis set. The denominator for percentages will be the number of subjects in the 
safety analysis set for each treatment or for all subjects as applicable. 
Medical history data will be listed by subject including visit, description of the disease/procedure, 
MedDRA system organ class (SOC), MedDRA preferred term (PT), start date, and stop date (or 
ongoing if applicable). 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 54 of 66 
09 May 2022 
 
9.5. 
Prior and Concomitant Medication and Drug Administration 
Prior medications are those that started and stopped prior to the first dose of study medication. 
Concomitant medications are those taken after first dosing (including medications that started prior 
to dosing and continued after). 
Prior and concomitant medication will be listed by subject and will include the following 
information: reported name, PT, the route of administration, dose, frequency, start date/time, 
duration and indication. 
Prior and concomitant medication will be coded according to the World Health Organization Drug 
Dictionary (WHO-DD) latest version. 
Drug administration dates and times will be listed for each subject. 
9.6. 
Exposure 
A listing of drug administration will be created and will include the date and time of administration. 
9.7. 
Safety Analyses 
Where applicable, pre-dose measurements on Day 1 are considered baseline values for safety 
variables. 
9.7.1. 
Adverse Events 
All AEs will be listed and summarized per treatment arm. The number and percent of subjects 
experiencing an event will be tabulated for each SOC and PT. The AEs will also be tabulated 
according to intensity and causality. A TEAE is defined as an AE with a start date on or after the 
first dose of study drug, or that worsened following first administration of study drug. For purposes 
of data analysis, the end of the TEAE period will be the last time point when a subject was 
evaluated for AEs. 
The AEs (TEAEs) will be tabulated according to intensity and causality, as applicable. 
Non-TEAEs will be listed, as applicable. 
Serious AEs will be listed separately. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 55 of 66 
09 May 2022 
 
9.7.2. 
Clinical Laboratory Tests 
Individual data listings of laboratory results will be presented for each subject. Flags will be 
attached to values outside of the laboratory’s reference limits along with the Principal 
Investigator’s assessment or designee. Clinically significant laboratory test abnormalities that were 
considered AEs by the Principal Investigator will be presented in the AE listings. 
Clinical laboratory tests (observed values) will be summarized descriptively in tabular format. 
Shift tables will be presented for select laboratory parameters. 
9.7.3. 
Vital Signs 
Individual data listings of vital signs (observed and change from baseline) will be presented for 
each subject. Individual clinically significant vital signs findings that were considered AEs by the 
Principal Investigator will be presented in the AE listings. 
Observed values as well as change from baseline data will be summarized descriptively in tabular 
format. 
9.7.4. 
Standard 12-lead Electrocardiogram 
Standard 12-lead ECG data (observed and change from baseline) will be listed for each subject 
and time point. Observed values will be summarized descriptively in tabular format. Change from 
baseline will be summarized descriptively for QTc data. A categorical QTc analysis will also be 
performed. 
9.7.5. 
Physical Examination 
Abnormal physical examination findings will be listed. 
9.8. 
Exploratory analysis 
Exploratory analysis may be presented separately from the main clinical study report (CSR). 
Further details on exploratory analyses will be provided in the SAP. 
9.9. 
Determination of Sample Size 
The determination of sample size is not based on statistical considerations of power but 25 subjects 
is considered sufficient to meet the study objectives. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 56 of 66 
09 May 2022 
 
10. 
ETHICAL, LEGAL AND ADMINISTRATIVE ASPECTS 
10.1. Data Quality Assurance 
The Sponsor or designee will conduct a study initiation visit to verify the qualifications of the 
Principal Investigator, inspect the facilities and inform the Principal Investigator of responsibilities 
and procedures for ensuring adequate and correct documentation. 
The Principal Investigator must prepare and maintain adequate and accurate records of all 
observations and other data pertinent to the clinical study for each study participant. Frequent 
communication between the clinical site and the Sponsor is essential to ensure that the safety of 
the study is monitored adequately. The Principal Investigator will make all appropriate safety 
assessments on an ongoing basis. The Medical Monitor may review safety information as it 
becomes available throughout the study. 
All aspects of the study will be carefully monitored with respect to Good Clinical Practice (GCP) 
and SOPs for compliance with applicable government regulations. The Study Monitor will be an 
authorized individual designated by the Sponsor. The Study Monitor will have access to all records 
necessary to ensure integrity of the data and will periodically review the progress of the study with 
the Principal Investigator. 
10.2. Data Collection and Access to Source Data/Documents 
The Principal Investigator will ensure the accuracy, completeness and timeliness of the data 
reported to the Sponsor. The Investigator or designee will cooperate with the Sponsor’s 
representative(s) for the periodic review of study documents to ensure the accuracy and 
completeness of the data at each scheduled monitoring visit. Data clarification requests will be 
provided to the study team by means of electronic or manual queries. 
The Investigator will allow Sponsor representatives, contract designees, authorized regulatory 
authority inspectors and the IEC to have direct access to all electronic records pertaining to the 
study. 
All data should be recorded, handled and stored in a way that allows its accurate reporting, 
interpretation and verification. 
10.3. Archiving Study Documents 
All source documents generated in connection with the study will be retained in the limited access 
file storage area, respecting the privacy and confidentiality of all records that could identify the 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 57 of 66 
09 May 2022 
 
subjects. Direct access is allowed only for authorized people for monitoring and auditing purposes. 
Source documents will be handled, stored and archived according to in-house procedures. 
The Investigator's Site File will be archived by the CRO for at least 2 years after completion of the 
study or longer if agreed upon by the Sponsor. 
10.4. Good Clinical Practice 
The procedures set out in this clinical study protocol are designed to ensure that the Sponsor and 
the Principal Investigator abide by the principles of the ICH guidelines on GCP. The clinical study 
also will be carried out in keeping with national and local legal requirements (in accordance with 
United States investigational new drug [IND] regulations [21 CFR 56]). 
10.5. Informed Consent 
Eligible subjects may only be included in the study after providing IRB/IEC approved informed 
consent. 
Informed consent must be obtained from the subject before conducting any study-specific 
procedure. 
As part of the informed consent procedure, the Principal Investigator or designee must explain 
orally and in writing the nature, duration and purpose of the study and the action of the drug in 
such a manner that the subject is aware of the potential risks, inconveniences or AEs that may 
occur. The subject should be informed that he/she is free to withdraw from the study at any time. 
Subjects will receive all information that is required by federal regulations and ICH guidelines. 
The ICF must be signed and dated; one copy will be handed to the subject, and the Principal 
Investigator will retain a copy as part of the clinical study records. The Principal Investigator will 
not undertake any investigation specifically required for the clinical study until written consent 
has been obtained. The terms of the consent and when it was obtained must be documented in the 
subject source documents. 
The Sponsor will review the Investigator-proposed ICF to ensure it complies with the ICH GCP 
guideline (including the ethical principles that have their origins in the Declaration of Helsinki) 
and regulatory requirements and is considered appropriate for this study. The Principal Investigator 
or designee will provide the Sponsor with a copy of the IRB/IEC approved ICF prior to the start 
of the study. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 58 of 66 
09 May 2022 
 
If a protocol amendment is required, then the ICF may need to be revised to reflect the changes to 
the protocol. If the ICF is revised, it must be reviewed and approved by the responsible IRB/IEC 
and signed by all subjects subsequently enrolled in the clinical study, as well as those currently 
enrolled in the clinical study as applicable. 
10.6. Insurance and Compensation for Injury 
The Sponsor has covered this clinical study by means of an insurance of the clinical study 
according to national requirements. The name and address of the relevant insurance company, the 
certificate of insurance, the policy number and the sum insured are provided in the Investigator’s 
Site File. 
10.7. Protocol Approval and Amendment(s) 
Before the start of the clinical study, the clinical study protocol and other relevant documents will 
be approved by the IRB/IEC, in accordance with local legal requirements. The Sponsor must 
ensure that all ethical and legal requirements have been met before the first subject is enrolled in 
the clinical study. 
This protocol is to be followed exactly. To alter the protocol, amendments must be written, which 
must be released by the responsible staff and receive IRB/IEC approval prior to implementation 
(as appropriate). 
Administrative changes may be made without the need for a formal amendment but will also be 
mentioned in the integrated CSR. All amendments will be distributed to all study protocol 
recipients, with appropriate instructions. 
10.8. Confidentiality Data Protection 
All clinical study findings and documents will be regarded as confidential. Study documents 
(protocols, IBs and other material) will be stored appropriately to ensure their confidentiality. The 
Principal Investigator and members of his/her research team (including the IRB/IEC) must not 
disclose such information without prior written approval from the Sponsor, except to the extent 
necessary to obtain informed consent from subjects who wish to participate in the study or to 
comply with regulatory requirements. 
The anonymity of participating subjects must be maintained. Subjects will be specified on study 
documents by their subject number, initial or birth date, not by name. Documents that identify the 
subject (e.g., the signed ICF) must be maintained in confidence by the Principal Investigator. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 59 of 66 
09 May 2022 
 
10.9. Publication Policy 
By signing the clinical study protocol, the Principal Investigator agrees with the use of results of 
the clinical study for the purposes of national and international registration, publication and 
information for medical and pharmaceutical professionals. If necessary, the regulatory authorities 
will be notified of the Principal Investigator’s name, address, qualifications and extent of 
involvement. 
A Principal Investigator shall not publish any data (poster, abstract, paper, etc.) without having 
consulted with the Sponsor in advance. 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 60 of 66 
09 May 2022 
 
11. 
REFERENCE LIST 
Avau, B., A. Rotondo, T. Thijs, et al. 2015. 'Targeting extra-oral bitter taste receptors 
modulates gastrointestinal motility with effects on satiation', Sci Rep, 5: 15985. 
Bray, G. A., G. Fruhbeck, D. H. Ryan, et al. 2016. 'Management of obesity', Lancet, 387: 
1947-56. 
Chen, M. C., S. V. Wu, J. R. Reeve, Jr., et al. 2006. 'Bitter stimuli induce Ca2+ signaling and 
CCK release in enteroendocrine STC-1 cells: role of L-type voltage-sensitive Ca2+ 
channels', Am J Physiol Cell Physiol, 291: C726-39. 
Collaborators, G. B. D. Obesity, A. Afshin, M. H. Forouzanfar, et al. 2017. 'Health Effects of 
Overweight and Obesity in 195 Countries over 25 Years', N Engl J Med, 377: 13-27. 
Deloose, E., P. Janssen, M. Corsetti, et al. 2017. 'Intragastric infusion of denatonium 
benzoate attenuates interdigestive gastric motility and hunger scores in healthy female 
volunteers', Am J Clin Nutr, 105: 580-88. 
Guh, D. P., W. Zhang, N. Bansback, et al. 2009. 'The incidence of co-morbidities related to 
obesity and overweight: a systematic review and meta-analysis', BMC Public Health, 
9: 88. 
Hruby, A., and F. B. Hu. 2015. 'The Epidemiology of Obesity: A Big Picture', 
Pharmacoeconomics, 33: 673-89. 
Janssen, S., J. Laermans, P. J. Verhulst, et al. 2011. 'Bitter taste receptors and alpha-gustducin 
regulate the secretion of ghrelin with functional effects on food intake and gastric 
emptying', Proc Natl Acad Sci U S A, 108: 2094-9. 
Kim, K. S., J. M. Egan, and H. J. Jang. 2014. 'Denatonium induces secretion of glucagon-like 
peptide-1 through activation of bitter taste receptor pathways', Diabetologia, 57: 
2117-25. 
Rodgers, R. J. 2017. 'Bench to bedside in appetite research: Lost in translation?', Neurosci 
Biobehav Rev, 76: 163-73. 
Rosa-Goncalves, P., and D. Majerowicz. 2019. 'Pharmacotherapy of Obesity: Limits and 
Perspectives', Am J Cardiovasc Drugs, 19: 349-64. 
Schier, L. A., T. L. Davidson, and T. L. Powley. 2011. 'Ongoing ingestive behavior is rapidly 
suppressed by a preabsorptive, intestinal "bitter taste" cue', Am J Physiol Regul Integr 
Comp Physiol, 301: R1557-68. 
Sibert, J. R., and N. Frude. 1991. 'Bittering agents in the prevention of accidental poisoning: 
children's reactions to denatonium benzoate (Bitrex)', Arch Emerg Med, 8: 1-7. 
 
 
 


Aar d var k T hera pe utics, I nc.  
                                             Cli nical St u d y Pr ot oc ol Versi o n 6 . 0 
A A R D -2 0 1  
C O N FI D E N TI A L  
 
 
Pa ge 6 1  of 6 6  
0 9  Ma y  2 0 2 2  
 
1 2.  
A P P E N DI X  
Pr ot oc ol: A A R D -2 0 1  
M M T T
 
S u bject #: _ _ _ _ _ _ _ _ _ _ _ _ _  
I R B # 2 1 0 2 1 0 
 
PI: Jere m y Pett us, M D  
Date: _ _ _ _ / _ _ _ _ _ _ / _ _ _ _ _ _ _ _  
S u bject I niti als:    _ _ _   _ _ _   _ _ _  
 
B o d y C o m p ositi o n  
Ti me: _ _ _ _ _ _ _ _ _ _ _ _  
Hei g ht: _ _ _ _ _ _ _ _ c m                 Wei g ht: _ _ _ _ _ _ _ _ _ _ k g                B MI: _ _ _ _ _ _ _ _  
 
 
B o d y Fat Perce nta ge: _ _ _ _ _ _ _ %  
St u d y Staff  I nitials a n d Date: _ _ _ _ _ _ _ _ _ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
Vit al Si g ns  
B P a n d He art R ate will be me as ure d i n t he sitti n g p ositi o n. T he s u bject M U S T be reste d f or at le ast 5 mi n.  
 
Ti me S u bject Starts Resti n g: _ _ _ _ _ _ _ _ _ _: _ _ _ _ _ _ _ _ _ _ _  
 
Ar m (circle):       L E F T           RI G H T          
 
Ti me: _ _ _ _ _ _ _ _ _ _ _        Bl o o d Press ure: _ _ _ _ _ _ _ _ _ / _ _ _ _ _ _ _ _ _ _ _ m m H g         Heart Rate: _ _ _ _ _ _ _ b p m  
 
A n y re pe at me as ure me nts of b ot h t he B P a n d he art r ate m ust be t a ke n at le ast 2 mi n . a p art a n d rec or de d. 
 
Ti me: _ _ _ _ _ _ _ _ _ _ _        Bl o o d Press ure:  _ _ _ _ _ _ _ _ _ / _ _ _ _ _ _ _ _ _ _ _ m m H g         Heart Rate: _ _ _ _ _ _ _ b p m  
Res pir at or y R ate :  _ _ _ _ _ _ _ _ _ breat hs/ mi n ute                                    B o d y Te m per at ure : _ _ _ _ _ _ _ _ _ _ _ _ ○C  
St u d y Staff I nitials a n d Date: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
A E/ S A Es & C o nc o mit a nt Me dic ati o ns  
Di d s u bject ha v e a n y c h a n ges t o t heir c urre nt me dicati o ns?  
*If yes, n ote c h a n ges o n t he c o nc o mit a nt me dic ati o n l o g.  
 Y E S *    N O  
Di d s u bject re p ort a n y a d verse e v e nts or st u d y relate d pr o ble ms? 
*If yes, n ote c h a n ges o n t he A E/ S A E  l o g. 
 Y E S *    N O  
St u d y Staff  I nitials a n d Date: _ _ _ _ _ _ _ _ _ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
 
 


Aar d var k T hera pe utics, I nc.  
                                             Cli nical St u d y Pr ot oc ol Versi o n 6 . 0 
A A R D -2 0 1  
C O N FI D E N TI A L  
 
 
Pa ge 6 2  of 6 6  
0 9  Ma y  2 0 2 2  
 
 
B aseli ne  
Di d s u bject f ast 1 0 h o urs as i nstr ucte d ?  
                                                                                                                             
Date of last meal:      _ _ _ _ _/ _ _ _ _ _ _ _ _/ _ _ _ _ _ _ Ti me of last meal:   _ _ _ _ : _ _ _ _                 Y E S      N O  
I nsert I V dr a w li ne i nt o a peri p her al ar m vei n a p pr o xi m atel y o ne h o ur bef ore first 
dr a w.  
St u d y Staff  I nitials a n d Date: _ _ _ _ _ _ _ _ _ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
Mi xe d Me al T oler a nce Test ( M M T T)  
S C H E D U L E D 
Ti m e p oi nt  
T ar g et 
ti m e  
( h h: m m) 
A C T U A L 
TI M E  
( h h: m m) 
S a m ple(s) c ollecte d / 
T as ks  
Y SI Gl uc ose 
re a di n g  
( at e ac h ti me p oi nt)  
 
Y SI # _ _ _ _ _  
N urse 
i niti als 
-1 2 0 mi n  
 
 
I nstr uct s u bject t o e m pt y bl a d der. 
 
 
-8 0 mi n  
 
 
St art I D C # 1:  
 
_ _ _ _ _ _ _ _ _ _ _ _ _  
 
 
-5 0 mi n  
 
 
St o p I D C # 1:  
 
_ _ _ _ _ _ _ _ _ _ _ _ _  
 
 
-3 0 mi n  
 
 
St u d y Dr u g Gi ve n I F S E C O N D M M T T  
 
Ti m e: _ _ _ _ _ _ _ _ _ _ _  
 
 
-1 5 mi n  
 
 
⃝ Gl uc os e  
⃝ F F A  
⃝ C C K  
⃝ GI P  
 
 
⃝ I ns uli n 
⃝ Gl uc a g o n  
⃝ P Y Y  
⃝ G L P -1  
⃝ C -Pe pti de  
⃝ G hreli n  
⃝ A m yli n  
 
 
-5 mi n  
 
 
⃝ Gl uc ose  
⃝ I ns uli n 
⃝ C -Pe pti de  
 
 
⃝ C yt o ki ne p a nel  
⃝
 Bile Aci ds/ Ferriti n/ C R P  
⃝ Le pti n/ A di p o necti n  
⃝  6 ml Ser u m st or a ge          
⃝  6 ml Pl as m a  st or a ge          
0 mi n  
 
 
Gi v e E ns ure li q ui d me al  
( 2 x E ns ure) 
C o ns u m e wit hi n 5 mi n utes  
 
St art Ti m e: _ _ _ _ _ _ _ _ _ _  
 
St o p Ti me: _ _ _ _ _ _ _ _ _ _ _  
 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 63 of 66 
09 May 2022 
 
15 min 
 
 
⃝ Glucose 
⃝ Insulin 
⃝ C-Peptide 
 
 
⃝  4 ml Serum storage         ⃝  4 ml Plasma storage       
30 min 
 
 
⃝ Glucose 
⃝ Insulin 
⃝ C-Peptide 
 
 
 
⃝  4 ml Serum storage         ⃝  4 ml Plasma storage       
60 min 
 
 
⃝ Glucose 
⃝ Insulin 
 
 
⃝ C-Peptide 
⃝ FFA 
⃝  4 ml Serum storage         ⃝  4 ml Plasma storage       
SCHEDULED 
Timepoint 
Target 
time  
(hh:mm) 
ACTUAL 
TIME 
(hh:mm) 
Sample(s) collected/ 
Tasks 
YSI Glucose 
reading 
(at each time 
point) 
 
YSI#_____ 
Nurse 
initials 
90 min 
 
 
⃝ Glucose 
⃝ Insulin 
⃝ C-Peptide 
 
 
 
⃝  4 ml Serum storage         ⃝  4 ml Plasma storage       
120 min 
 
 
 
⃝ Glucose 
⃝ FFA 
⃝ CCK 
⃝ GIP 
 
 
 
⃝ Insulin 
⃝ Glucagon 
⃝ PYY 
⃝ GLP-1 
⃝ C-Peptide 
⃝ Ghrelin 
⃝ Amylin 
⃝  6 ml Serum storage         ⃝  6 ml Plasma storage       
150 min 
 
 
⃝ Glucose 
⃝ Insulin 
⃝ C-Peptide 
 
 
 
⃝  4 ml Serum storage          
⃝  4 ml Plasma storage    
180 min 
 
 
⃝ Glucose 
⃝ FFA 
 
 
⃝ Insulin 
⃝ C-Peptide 
⃝  4 ml Serum storage          
⃝  4 ml Plasma storage    
195 min 
 
 
Start IDC #2: 
 
_____________ 
 
 
210 min 
 
 
⃝ Glucose 
⃝ Insulin 
⃝ C-Peptide 
 
 
 
⃝  4 ml Serum storage        ⃝  4 ml Plasma storage   
225 min 
 
 
Stop IDC #2: 
 
_____________ 
 
 


Aar d var k T hera pe utics, I nc.  
                                             Cli nical St u d y Pr ot oc ol Versi o n 6 . 0 
A A R D -2 0 1  
C O N FI D E N TI A L  
 
 
Pa ge 6 4  of 6 6  
0 9  Ma y  2 0 2 2  
 
2 4 0 mi n  
 
 
⃝ Gl uc os e  
⃝ F F A  
⃝ C C K  
⃝ GI P  
 
 
 
 
⃝ I ns uli n 
⃝ Gl uc a g o n  
⃝ P Y Y  
⃝ G L P -1  
⃝ C -Pe pti de  
⃝ G hreli n  
⃝ A m yli n  
⃝  4  ml Ser u m st or a ge          
⃝  4  ml Pl as m a  st or a ge          
A F T E R  fi n al bl o o d dr a w, re m o ve I V. 
N urse c o m pleti n g pr oce d ure Si g nat ure a n d Date: 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
Atte n di n g P h ysicia n ( PI/ S u b -I) Si g nat ure a n d Date: 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
P ost -Pr oce d ure  
P ost pr oce d ure m e al e ate n ?                      Y E S      N O          Ti m e: _ _ _ _ _ _ _ _ _ _ _  
P ost -Pr o ce d ure Gl uc ose  C hec k ?                Y E S      N O          Ti m e: _ _ _ _ _ _ _ _ _ _ _           B G: _ _ _ _ _ _ _ _ _  
m g/ d L  
 
                                                                                                            Ti m e: _ _ _ _ _ _ _ _ _ _ _           B G: 
_ _ _ _ _ _ _ _ _  m g/ d L  
A n y A d verse E ve nts t o re p ort d uri n g o r after st u d y pr oce d ure ?  Y E S *    N O  
*If yes, ple ase descri be bel o w:  
 
 
St u d y Staff  I nitials a n d Date: _ _ _ _ _ _ _ _ _ __ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
 
 


Aar d var k T hera pe utics, I nc.  
                                             Cli nical St u d y Pr ot oc ol Versi o n 6 . 0 
A A R D -2 0 1  
C O N FI D E N TI A L  
 
 
Pa ge 6 5  of 6 6  
0 9  Ma y  2 0 2 2  
 
P ost -pr oce d ure Vit al Si g ns  
B P a n d He art R ate will be me as ure d i n t he sitti n g p ositi o n. T he s u bject M U S T be reste d f or at le ast 5 mi n.  
 
Ti me S u bject Starts Resti n g: _ _ _ _ _ _ _ _ _ _: _ _ _ _ _ _ _ _ _ _ _  
 
Ar m (circle):       L E F T           RI G H T          
 
Ti me: _ _ _ _ _ _ _ _ _ _ _        Bl o o d Press ure: _ _ _ _ _ _ _ _ _ / _ _ _ _ _ _ _ _ _ _ _ m m H g         Heart Rate: _ _ _ _ _ _ _ b p m  
 
A n y re pe at me as ure me nts of b ot h t he B P a n d he art r ate m ust be t a ke n at le ast 2 mi n . a p art a n d rec or de d. 
 
Ti me: _ _ _ _ _ _ _ _ _ _ _        Bl o o d Press ure:  _ _ _ _ _ _ _ _ _ / _ _ _ _ _ _ _ _ _ _ _ m m H g         Heart Rate: _ _ _ _ _ _ _ b p m  
Res pir at or y R ate :  _ _ _ _ _ _ _ _ _ breat hs/ mi n ute                                    B o d y Te m per at ure : _ _ _ _ _ _ _ _ _ _ _ _ ○C  
St u d y Staff I nitials a n d Date: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
Ne xt Visit  
 
N E X T VI SI T :  _ _ _ _ _/ _ _ _ _ _ _ _ _ _/ _ _ _ _ _ _ _ _ _ @ _ _ _ _ _ _: _ _ _ _ _ _ _ _  
W as  s u bject gi ve n i nstr ucti o ns f or ne xt visit ? 
 Y E S   N O  
St u d y Staff I nitials a n d Date: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
N otes  
 
 
 
 
 
 
 
 


Aardvark Therapeutics, Inc. 
                                             Clinical Study Protocol Version 6.0 
AARD-201 
CONFIDENTIAL 
 
 
Page 66 of 66 
09 May 2022 
 
 
 
 
 
 
 
 
 
 
 
 
Study Staff Initials and Date: _____________________________________