Clinical trial NCT04705415
Conditions: Healthy
Sponsor: NeuroBo Pharmaceuticals Inc.
Start date: 2020-11-17; Completion date: 2022-10-28
Enrollment: 54 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
A single and multiple ascending dose study of ANA001 in healthy adults to assess the safety and pharmacokinetics

Detailed description:
This is a Phase 1, single center, randomized, double blind, single ascending dose (SAD) and multiple ascending dose (MAD) study to assess the safety and pharmacokinetics of ANA001 in healthy adult subjects. In the single ascending dose portion of the study, subjects in 3 cohorts of 10 subjects each will be randomized to receive a single daily oral dose of ANA001 or matching placebo. In the multiple ascending dose portion of the study, subjects in 3 cohorts of 12 subjects each will be randomized to receive twice or thrice daily oral dose of ANA001 or matching placebo.

Arms: arm 1: Subjects will receive a 1000 mg single oral dose of ANA001 with a standardized light meal presented at 4 x 250 mg capsules. Each capsule is 250 mg. arm 2: Subjects will a 2000 mg single oral dose of ANA001 with a standardized light meal presented at 8 x 250 mg capsules. Each capsule is 250 mg. arm 3: Subjects will a 3000 mg single oral dose of ANA001 with a standardized light meal presented at 12 x 250 mg capsules. Each capsule is 250 mg. arm 4: Subjects will receive matching placebo hydroxypropylmethylcellulose (HPMC) as a single oral dose (4, 8, or 12 capsules) with a standardized light meal. arm 5: Subjects will receive an oral dose of 1000mg ANA001 twice daily (BID) with a standardized light meal for 7 consecutive days. Each dose will be presented as 4 250mg capsules. arm 6: Subjects will receive an oral dose of 1000mg ANA001 three times daily (TID) with a standardized light meal for 7 consecutive days. Each dose will be presented as 4 250mg capsules. arm 7: Subjects will receive an oral dose of matching Placebo to ANA001 two (BID) three times daily (TID) with a standardized light meal for 7 consecutive days. Each dose will be presented as 4 250mg capsules of hydroxypropylmethylcellulose (HPMC)
Interventions: intervention 1: Niclosamide intervention 2: Placebo
Intervention descriptions: intervention 1: Niclosamide is an antihelmintic with in-vitro antiviral activity intervention 2: Matching hydroxypropylmethylcellulose HPMC capsules with no active ingredients

Protocol documents:
Prot_000.pdf: 
CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 1 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Title Page 
Protocol Title:  
A Randomized, Double-Blind, Single and Multiple Ascending Dose Study to Assess the Safety 
and Pharmacokinetics of ANA001 in Healthy Adults 
Protocol Number: ANA001-002.02 
Compound Number: ANA001 
Short Title:  
A single ascending dose (SAD) and multiple ascending dose (MAD) PK study of ANA001in 
healthy adults 
Sponsor Name and Legal Registered Address:  
ANA Therapeutics, Inc.  
602 Bainbridge St. 
Foster City, CA 94404 USA 
 
Regulatory Agency Identifying Number(s):  
IND #148459 
 
Protocol Finalization Date: 09 January 2021 
Version: 2.0 
 
 
 
Confidentiality Statement 
The information in this document contains trade secrets and commercial information that are privileged or confidential and may not be disclosed 
unless such disclosure is required by applicable law or regulations. In any event, persons to whom the information is disclosed must be informed 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 2 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
that the information is privileged or confidential and may not be further disclosed by them. These restrictions on disclosure will apply equally to 
all future information supplied to you that is indicated as privileged or confidential. 
 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 3 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
INVESTIGATOR AGREEMENT 
This protocol was designed and will be conducted, recorded, and reported in compliance with the 
principles of Good Clinical Practice (GCP) guidelines.  These guidelines are stated in U.S. 
federal regulations, as well as the “Guideline for Good Clinical Practice,” International 
Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals 
for Human Use.  
I have read this protocol and agree that it contains all necessary details for carrying out this 
study. I will conduct the study as outlined herein and will complete the study within the time 
designated. 
I will provide copies of the protocol and all pertinent information to all individuals responsible to 
me who assist in the conduct of the study. I will discuss this material with them to ensure they 
are fully informed regarding the study drug and the conduct of the study. 
Principal Investigator 
Name: 
Helen Paguntalan, MD 
Institution and Address: 
WCCT Global 
5630 Cerritos Ave. 
Cypress, CA 90630 
Telephone number: 
714-252-0700 
Signature: __________________________________________________________ Date: _____________________ 
 
 
 
 
 
 
 
 
 
 
(Day Month Year) 
 
Sponsor’s Responsible Medical Officer: 
Name: 
Doug Rank, MD, Vice President of Clinical Development 
Institution and Address: 
ANA Therapeutics, Inc. 
602 Bainbridge St. 
Foster City, CA 94404 USA 
Telephone number: 
415-990-4073 
Signature: __________________________________________________________ Date: _____________________ 
 
 
 
 
 
 
 
 
 
 
(Day Month Year) 
 
 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 4 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
STUDY ADMINISTRATION  
Study Role 
Name, title, contact information 
Sponsor 
ANA Therapeutics, Inc. 
602 Bainbridge St. 
Foster City, CA 94404 USA 
Principal Investigator  
Helen Paguntalan, MD 
WCCT Global 
5630 Cerritos Ave. 
Cypress, CA 90630 USA 
Medical Monitor 
Doug Rank, MD 
602 Bainbridge St. 
Foster City, CA 94404 USA 
Biostatistics and 
Pharmacokinetics 
Kalyan Ghosh, PhD 
Inference, Inc. 
701 Lee Rd. 
Chesterbrook, PA 19087 USA 
Clinical laboratory  
Innovative Bioanalysis, LLC 
5630 Cerritos Ave. 
Cypress, CA 90630 USA 
Bioanalytical laboratory  
(PK-analysis) 
Medpace Bioanalytical Laboratories 
5375 Medpace Way 
Cincinnati, Ohio 45227 USA 
Tel: 513-579-9911 
Fax: 513-579-044 
 
 
 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 5 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Protocol Amendment History 
Date 
Description 
24-Dec-20 
Amendment 1 
 
The protocol is amended to address comments received by the FDA as follows: 
 
- 
Revising the Inclusion criteria pulse rate limits from 40 -100 bpm to 50 – 100 bpm 
 
- 
Providing reference laboratory values that will be used to determine if laboratory results are considered 
clinically significant 
 
- 
Providing a standard toxicity grading scale 
 
- 
Providing of detailed individual and dose level stopping criteria based upon the known safety profile and 
mechanism of action of ANA001 (niclosamide) 
 
- 
Limiting the total daily dose in the MAD portion of the protocol to 2000 mg 
 
Page 8. MAD 
Total daily doses are anticipated to range between will not exceed 2000 mg and 4500 mg of ANA001.  
 
Page 18. Section 3.3 Benefit/Risk Assessment  
The current SAD/MAD study is designed to explore the safety and PK of dosing greater than up to 2000 3000 
mg/day as a single dose and regimens not to exceed 2000 mg/day as divided doses BID or TID. 
 
Page 20. MAD 
Total daily doses are anticipated to range between will not exceed 2000 mg and 4500 mg of ANA001.  
 
Page 21. Section 5.5 Justification for Dose 
The current study is designed to assess the safety of single doses of ANA001 exceeding up to 3000 2,000 mg 
and multiple dosing regimens up to not to exceed 2000 mg daily divided either BID or TID.  


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 6 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Date 
Description 
24-Dec-20 
Amendment 1 
 
Page 22. Section 6.1 Inclusion Criteria 
 
6. Pulse rate between 40 50 and 100 beats per minute (bpm) 
 
Page 23. Section 6.2 Exclusion Criteria 
Clinically significant abnormal values for hematology, clinical chemistry, or urinalysis at screening as deemed 
appropriate by the Investigator assessed per Appendix 6. 
 
Page 25. 7.2 Dose Modification 
 
Following review of the safety and PK data of three SAD cohorts, a fourth cohort may be added, not to exceed 
4500 mg. 
Total daily doses are anticipated to be between will not exceed 2000 mg and 4500 mg. 
 
Page 31. Section 9.1.4 Clinical Safety Laboratory Assessments 
The Investigator must review the laboratory report, document this review, and record any clinically relevant 
changes (as per the reference laboratory values [Appendix 6]) occurring during the study in the AE section of 
the CRF. 
All laboratory tests with values considered clinically significantly abnormal during participation in the study 
should be repeated until the values return to normal or baseline or are no longer considered clinically significant 
by the Investigator or medical monitor as per the reference laboratory values in Appendix 6. 
If laboratory values from non-protocol specified laboratory assessments performed at the institution’s local 
laboratory require a change in participant management or are considered clinically significant by the Investigator 
as given in Appendix 6 (e.g., SAE or AE or dose modification), then the results must be recorded in the CRF. 
 
Page 33.  Insertion of 9.3 Stopping Rules in Response to SAE 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 7 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Date 
Description 
24-Dec-20 
Amendment 1 
 
Dose-limiting toxicity AEs will be defined as Grade ≥3, treatment-emergent, treatment-related laboratory 
abnormalities (Appendix 6) or AEs per the grading system by US Department of Health and Human Services 
(CTCAE, Version 5.0). 
 
The following Stopping Criteria will be used: 
 
Clinical Findings: 
 
1. Anaphylaxis or Grade ≥3 skin rash or pruritis 
 
2. Grade ≥3 nausea, emesis or diarrhea 
 
Laboratory Findings: 
 
1. Evidence of new onset of acute renal or hepatic failure, defined by increases in BUN, creatinine, or LFTs 
(AST, ALT, Alkaline phosphatase) that reflect new evidence of Grade 3 thresholds for liver function tests, 
for creatinine, and for electrolytes attributable to the administration of drug in the context of the 
Investigator’s evaluation.  
 
2. Evidence of acute infection or inflammation, defined by changes in the WBC and differential indices, 
attributable to the administration of drug in the context of the Investigator’s evaluation.  
 
3. Any “unexpected” Grade ≥3 Laboratory test result.  
  
If one of the above clinical or laboratory criteria for stopping occurs in any subject, further study recruitment and 
subject dosing will be halted until a full safety assessment can be completed.  Unblinding of the affected subject 
will be performed and the PI and Sponsor will review the totality of clinical data and any other circumstances 
possibly related to the “Stopping Criteria” findings to determine if the study can be resumed, modified, or will 
be permanently terminated.  
 
Page 33. Section 9.3 Treatment of Overdose 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 8 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Date 
Description 
24-Dec-20 
Amendment 1 
For this study, any dose of ANA001 greater than 4500 3000 mg within a 24-hour time period will be considered 
an overdose. 
 
Page 37. Section 11 References 
Inserted reference in relation to Stopping rules: US Department of Health and Human Services. Common 
Terminology Criteria for Adverse Events (CTCAE). Version 5.0. Published: November 27, 2017. 
 
Page 50. Appendix 4 
Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) or other safety assessments 
(e.g., ECG, radiological scans, vital signs measurements), including those that worsen from baseline, considered 
clinically significant in the medical and scientific judgment of by the Investigator per Appendix 6 (i.e., not 
related to progression of underlying disease). 
 
Insertion of Appendix 6 Potentially Clinically Significant Laboratory Criteria 
 
Editorial Changes 
 
Language in bold has been added to the document for clarification purposes. Minor formatting and typographical errors 
were corrected throughout the protocol. 
 
Page 7. Section 1 Synopsis and Page 15. Section 3.1 Study Rationale 
ANA001 is a new formulation containing 250 mg niclosamide per capsule. Although other formulations 
containing niclosamide (ANA001) have been studied for tapeworm infections and other potential human uses, 
limited systemic PK/PD data exists in the literature. To ensure proper systemic levels of niclosamide are 
achieved for therapeutic effect, a SAD/MAD study with ANA001 capsules is necessary. 
 
Page 7. Section 1 Synopsis 
ANA001 capsule containing 250 mg niclosamide, administered orally as 4, 6, 8 or 12 capsules with a light meal 
(500 to 750 cal) or snack. 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 9 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Date 
Description 
24-Dec-20 
Amendment 1 
All participants on active treatment who contributed at least one post dose blood sample, evaluable for drug PK 
concentration will be included in the PK analysis. 
 
Global change throughout protocol 
Replaced ‘postdose’ with ‘after dosing’ and ‘predose’ with ‘before dosing’ expect with analysis timepoints. 
 
Page 14. Section 3 Introduction 
Due to its pleiotropic pharmacologic activity, niclosamide has been and is being continues to be studied in for a 
variety of other clinical indications. 
COVID-19 is results in a cytokine release syndrome. 
 
Page 43. Appendix 1 
Updated as per changes in text. 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 10 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Table of Contents 
 
Protocol Amendment History .......................................................................................................5 
1. 
Synopsis..........................................................................................................................13 
2. 
Schedule of Activities (SoA) .........................................................................................16 
3. 
Introduction ...................................................................................................................20 
3.1. 
Study Rationale ...............................................................................................................21 
3.2. 
Background .....................................................................................................................21 
3.2.1. 
Chemical Name/Structure ...............................................................................................22 
3.2.2. 
Nonclinical Studies .........................................................................................................22 
3.2.3. 
Safety ..............................................................................................................................23 
3.3. 
Benefit/Risk Assessment ................................................................................................24 
4. 
Objectives and Endpoints ............................................................................................25 
5. 
Study Design ..................................................................................................................26 
5.1. 
Overall Design ................................................................................................................26 
5.2. 
Participant and Study Completion ..................................................................................27 
5.3. 
End of Study Definition ..................................................................................................27 
5.4. 
Scientific Rationale for Study Design ............................................................................27 
5.5. 
Justification for Dose ......................................................................................................27 
6. 
Study Population ...........................................................................................................28 
6.1. 
Inclusion Criteria ............................................................................................................28 
6.2. 
Exclusion Criteria ...........................................................................................................29 
6.3. 
Lifestyle Restrictions ......................................................................................................30 
6.3.1. 
Meals and Dietary Restrictions .......................................................................................30 
6.3.2. 
Caffeine, Alcohol, and Tobacco .....................................................................................30 
6.3.3. 
Activity ...........................................................................................................................30 
6.4. 
Screen Failures ................................................................................................................30 
7. 
Treatments .....................................................................................................................31 
7.1. 
Treatments Administered ................................................................................................31 
7.2. 
Dose Modification ..........................................................................................................31 
7.3. 
Method of Treatment Assignment ..................................................................................31 
7.4. 
Blinding ..........................................................................................................................31 
7.5. 
Preparation/Handling/Storage/Accountability ................................................................32 
7.6. 
Treatment Compliance ....................................................................................................32 
7.7. 
Concomitant Therapy .....................................................................................................32 
7.7.1. 
Rescue Medicine .............................................................................................................33 
7.8. 
Treatment after the End of the Study ..............................................................................33 
8. 
Discontinuation/Withdrawal Criteria .........................................................................34 
8.1. 
Discontinuation of Study Treatment ...............................................................................34 
8.2. 
Withdrawal from the Study.............................................................................................34 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 11 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
8.3. 
Lost to Follow Up ...........................................................................................................34 
9. 
Study Assessments and Procedures.............................................................................35 
9.1. 
Safety Assessments .........................................................................................................36 
9.1.1. 
Physical Examinations ....................................................................................................36 
9.1.2. 
Vital Signs .......................................................................................................................37 
9.1.3. 
Electrocardiograms .........................................................................................................37 
9.1.4. 
Clinical Safety Laboratory Assessments ........................................................................37 
9.2. 
Adverse Events ...............................................................................................................38 
9.2.1. 
Time Period and Frequency for Collecting AE and SAE 
Information .....................................................................................................................38 
9.2.2. 
Method of Detecting AEs and SAEs ..............................................................................38 
9.2.3. 
Follow-up of AEs and SAEs ...........................................................................................38 
9.2.4. 
Regulatory Reporting Requirements for SAEs ...............................................................39 
9.2.5. 
Pregnancy ........................................................................................................................39 
9.3. 
Stopping Rules in Response to SAEs or Other Events ...................................................39 
9.4. 
Treatment of Overdose ...................................................................................................40 
9.5. 
Pharmacokinetics ............................................................................................................40 
9.6. 
Medical Resource Utilization and Health Economics ....................................................41 
10. 
Statistical Considerations .............................................................................................42 
10.1. 
Sample Size Determination ............................................................................................42 
10.2. 
Populations for Analyses ................................................................................................42 
10.3. 
Statistical Analyses .........................................................................................................42 
10.3.1. Safety Analyses ...............................................................................................................42 
10.3.2. Pharmacokinetic Analyses ..............................................................................................43 
10.3.3. Interim Analyses .............................................................................................................43 
11. 
References ......................................................................................................................44 
12. 
Appendices .....................................................................................................................49 
Appendix 1: Abbreviations and Terms ..........................................................................................50 
Appendix 2: Clinical Laboratory Tests ..........................................................................................53 
Appendix 3: Study Governance Considerations ............................................................................54 
Regulatory and Ethical Considerations .................................................................................54 
Financial Disclosure ..............................................................................................................54 
Informed Consent Process ....................................................................................................54 
Data Protection ......................................................................................................................55 
Dissemination of Clinical Study Data ...................................................................................55 
Data Quality Assurance ........................................................................................................56 
Source Documents ................................................................................................................57 
Study and Site Closure ..........................................................................................................57 
Appendix 4: Adverse Events: Definitions and Procedures for Recording, 
Evaluating, Follow-up, and Reporting ............................................................................58 
Appendix 5: Contraceptive Guidance and Collection of Pregnancy 
Information .....................................................................................................................64 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 12 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Appendix 6: Potentially Clinically Significant Laboratory Criteria ..............................................68 
  
List of Tables 
Table 1 
Schedule of Activities for SAD ......................................................................... 16 
Table 2 
Schedule of Activities for MAD ....................................................................... 17 
Table 3 
Schedule of PK Sampling for MAD ................................................................. 19 
Table 4 
 Protocol-Required Safety Laboratory Assessments ......................................... 53 
Table 5: 
 Highly Effective Contraceptive Methods ......................................................... 65 
 
List of Figures 
Figure 1 
Phylogenetic Tree Showing Antiviral Effects of Niclosamide Against Various 
Virus Families .................................................................................................... 21 
 
 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 13 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
1. 
Synopsis 
Protocol Title: A Randomized, Double-Blind, Single and Multiple Ascending Dose Study to 
Assess the Safety and Pharmacokinetics of ANA001 in Healthy Adults 
Short Title: A single ascending dose (SAD) and multiple ascending dose (MAD) PK study of 
ANA001 in healthy adults 
Rationale: 
ANA001 is a new formulation containing 250 mg niclosamide per capsule. Although other 
formulations containing niclosamide (ANA001) have been studied for tapeworm infections and 
other potential human uses, limited systemic PK/PD data exists in the literature. To ensure proper 
systemic levels of niclosamide are achieved for therapeutic effect, a SAD/MAD study with 
ANA001 capsules is necessary. 
Objectives and Endpoints 
Objectives 
Endpoints 
Primary 
 
• To assess safety and tolerability of 
single oral daily doses of ANA001 
(1000 mg, 2000 mg and 3000 mg) 
• To assess safety and tolerability of 
multiple daily oral doses (twice daily 
[BID] or thrice daily [TID]) of 
ANA001 for 7 days 
• Incidence of adverse events (AEs), 
serious AEs (SAEs), study drug 
discontinuation due to an AE (MAD 
portion only), use of concomitant 
medications, and change from baseline 
in clinical laboratory tests, vital signs, 
electrocardiograms (ECG), and 
physical examinations 
Secondary 
 
• To assess the PK of single and 
multiple doses of ANA001 
• AUC0-t, AUC0-last, AUC0-∞ (SAD 
only), Cmax, tmax, t1/2, CL/F, Vz/F 
Overall Design: 
This is a randomized, double-blind study to be conducted in two parts: single ascending dose 
(SAD) and multiple ascending dose (MAD). Potential participants for each part will undergo 
screening procedures within 30 days of enrolment.  
SAD 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 14 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
On Day -1, eligible participants will be randomly assigned to active drug or placebo in an 8:2 ratio. 
On Day 1, following an overnight fast of at least 10 hours, participants will receive their assigned 
treatment (1000 mg, 2000 mg, or 3000 mg of ANA001) with a standardized light meal (500 to 
750 cal).  A baseline ECG will be performed at Screening and at 3 hours after dosing. Blood 
pressure and heart rate will be measured every 8 hours while confined to the clinic. Blood for PK 
will be collected before dosing and at 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 hours after dosing. Participants 
will be discharged on Day 2 following the 24-hour PK sample and completion of clinical 
laboratory tests and physical examination. They will return on Day 7 (±2) for follow-up procedures 
including physical examination, laboratory tests, and assessment of AEs and concomitant 
medication use. 
MAD 
On Day -1, eligible participants will be randomly assigned to active drug or placebo in a 10:2 ratio 
on a BID (q12h) or TID (q8h) schedule, to be determined based on the PK results of the SAD 
cohorts. Total daily doses will not exceed 2000 mg of ANA001. On Days 1 through 7, participants 
will receive their assigned treatment in the morning with a standardized light meal (500 to 750 cal) 
following an overnight fast of at least 10 hours. Afternoon or evening doses will be given with a 
light snack. A baseline ECG will be performed at Screening and at 3 hours after the second dose 
on Day 7. Blood pressure and heart rate will be measured every 8 hours while confined to the 
clinic.  
During BID dosing, blood for PK will be collected on Days 1 and 7 before dosing and at 0.5, 1, 2, 
4, 6, 8, 10, and 12 hours after dosing. Blood samples will also be collected on Days 2, 4, 6, and 8 
before dosing and before the 12 hour dose on Days 2, 4, and 6.  
During TID dosing, blood for PK will be collected on Days 1 and 7 before dosing and at 0.5, 1, 2, 
4, 6, and 8 hours after dosing. Blood samples will also be collected on Days 2, 4, 6, and 8 before 
dosing, and before the 8 hour dose on Days 2, 4, and 6, and before the 16 hour dose on Day 6. 
Participants will be discharged on Day 8 following the last PK sample and completion of clinical 
laboratory tests and physical examination. They will return on Day 15 (±2) for follow-up 
procedures including physical examination, laboratory tests, and assessment of AEs and 
concomitant medication use. 
Number of Participants: 
A minimum of 30 participants will be enrolled in the SAD part and up to 36 participants in the 
MAD part. At least 3 participants of each sex will be included in each cohort. 
Treatment Groups and Duration: 
SAD 
Cohort S1: 1000 mg (n=8) or placebo (n=2), single dose 


CONFIDENTIAL 
 
ANA Therapeutics, Inc. 
Protocol ANA001-002.02 
Page 15 of 69 
 
Status: Final v2.0; Date 09Jan2021 
CONFIDENTIAL 
Cohort S2: 2000 mg (n=8) or placebo (n=2), single dose 
Cohort S3: 3000 mg (n=8) or placebo (n=2), single dose 
MAD 
Cohort M1: xx mg (n=9) or placebo (n=3) BID or TID for 7 days 
Cohort M2: xx mg (n=9) or placebo (n=3) BID or TID for 7 days 
Cohort M3: xx mg (n=9) or placebo (n=3) BID or TID for 7 days 
Investigational Product, Dose, and Mode of Administration 
ANA001 capsule containing 250 mg niclosamide, administered orally as 4, 6, 8 or 12 capsules 
with a light meal (500 to 750 cal) or snack. 
Statistical Methods 
Sample Size Determination: 
Sample size was not based on statistical considerations. 
Safety Analysis: 
All participants who received at least one dose of study drug will be included in the safety analysis. 
Safety will be assessed by the incidence and frequency of AEs, and changes in physical 
examinations, vital signs, clinical laboratory tests, and ECGs. Results will be summarized using 
descriptive statistics (mean, standard deviation [SD], coefficient of variation [%CV], median, 
minimum, maximum). 
Pharmacokinetic Analysis: 
All participants on active treatment who contributed at least one post dose blood sample, evaluable 
for drug PK concentration will be included in the PK analysis. PK parameters, calculated using 
non-compartmental models, include AUC0-∞, AUC0-t, AUC0-last, Cmax, tmax, t1/2, CL/F, and Vz/F. 
Results will be summarized using descriptive statistics (mean, standard deviation [SD], coefficient 
of variation [%CV], median, minimum, maximum). Plasma concentration-time profiles will be 
plotted. 
 


CONFIDENTIAL 
 
ANA Therapeutics 
Protocol ANA001 
Page 16 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
2. 
Schedule of Activities (SoA) 
Table 1 
Schedule of Activities for SAD 
  
Screening 
In-Patient 
Discharge Follow-
up 
Study Day 
 -30 to -2 
-1 
1 
2 
7±2 
Study Procedures 
  
COVID Pre-screen informed consent 
X 
  
  
  
  
Full study informed consent 
X 
  
  
  
  
Medical history 
X 
  
  
  
  
Prior medications 
X 
  
  
  
  
Physical examination  
X 
  
  
  
  
Abbreviated PE 
  
X 
X 
X 
X 
Viral Serology (HIV, HCV, HBV) 
X 
  
  
  
  
Urine alcohol and drug screen 
X 
X 
  
  
  
Inclusion / exclusion criteria 
X 
X 
  
  
  
Pregnancy test for WOCBP 
X 
X 
  
  
  
Randomization 
  
X 
  
  
  
CPU Admission & Confinement 
  
X 
X 
  
  
CPU Discharge 
  
  
  
X 
  
Drug Administrationa 
  
 
X 
  
  
Vital signsb 
X 
X 
X  
X 
X 
Height 
X 
  
  
  
  
Weight 
X 
X 
  
  
  
ECGc 
X 
X  
  
  
Hematology 
X 
X 
  
X 
X 


CONFIDENTIAL 
 
ANA Therapeutics 
Protocol ANA001 
Page 17 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
  
Screening 
In-Patient 
Discharge Follow-
up 
Study Day 
 -30 to -2 
-1 
1 
2 
7±2 
Study Procedures 
  
Serum chemistry 
X 
X 
  
X 
X 
Urinalysis 
X 
  
  
X 
X 
Pharmacokineticsd 
  
  
X 
X 
  
Adverse events 
  
Continuous 
Concomitant medications 
  
Continuous 
WOCBP = woman of child-bearing potential 
a.) Administered with Standardized light meal 
 
 
 
 
b.) Blood pressure and heart rate supine for 3 to 5 mins and every 8 hours during CPU confinement 
c.)  Supine for 5 mins at Screening and 3 hours after dosing 
 
d.) 4 mL before dosing and 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 hr after 
dosing 
 
 
 
 
Table 2 
Schedule of Activities for MAD 
  
Screening 
Confinement/Treatment Period 
Discharge 
Follow-up 
Study Day 
-30 to -2 
-1 
1 
2 
3 
4 
5 
6 
7 
8 
D15±2 
Study Procedures 
  
COVID Pre-screen ICF 
X 
  
  
  
  
  
  
  
  
  
  
Full study informed consent 
X 
  
  
  
  
  
  
  
  
  
  
Medical history 
X 
  
  
  
  
  
  
  
  
  
  
Prior medications 
X 
  
  
  
  
  
  
  
  
  
  
Physical examination  
X 
  
  
  
  
  
  
  
  
  
  
Abbreviated PE 
  
X 
  
X 
  
  
X 
  
  
X 
X 
Viral Serology (HIV, HCV, 
HBV) 
X 
  
  
  
  
  
  
  
  
  
  


CONFIDENTIAL 
 
ANA Therapeutics 
Protocol ANA001 
Page 18 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
  
Screening 
Confinement/Treatment Period 
Discharge 
Follow-up 
Study Day 
-30 to -2 
-1 
1 
2 
3 
4 
5 
6 
7 
8 
D15±2 
Study Procedures 
  
COVID Surveillance (ELISA) 
  
+ Swab 
  
  
X 
  
  
X 
  
  
  
Urine alcohol and drug screen 
X 
X 
  
  
  
  
  
  
  
  
  
Pregnancy test for WOCBP 
X 
X 
  
  
  
  
  
  
  
  
  
Inclusion / exclusion criteria 
X 
X 
  
  
  
  
  
  
  
  
  
Randomization 
  
X 
  
  
  
  
  
  
  
  
  
CPU Admission & Confinement 
  
X 
X 
X 
X 
X 
X 
X 
X 
  
  
CPU Discharge 
  
  
  
  
  
  
  
  
  
X 
  
Drug Administrationa 
  
  
X 
X 
X 
X 
X 
X 
X 
  
  
Vital signsb 
X 
X 
X  
X 
X 
X 
X 
X 
X 
X 
X 
Height 
X 
  
  
  
  
  
  
  
  
  
  
Weight 
X 
  
  
  
  
  
  
  
  
  
  
ECGc 
X 
  
  
  
  
  
  
  
 X 
  
  
Hematology 
X 
X 
  
X 
  
  
X 
  
  
X 
X 
Serum chemistry 
X 
X 
  
X 
  
  
X 
  
  
X 
X 
Urinalysis 
X 
  
  
X 
  
  
X 
  
  
X 
X 
Pharmacokineticsd 
  
  
X 
X 
X 
X 
X 
X 
X 
X 
  
Adverse events 
  
Continuous 
Concomitant medications 
  
Continuous 
WOCBP = woman of child-bearing potential 
a.) Administered with standardized light meal (500 to 750 cal) in AM and light snack (PM) if BID (q12h) or TID (q8h) daily on days 1 through 6 
and a single AM dose on day 7 
b.) Blood pressure and heart rate supine for 3 to 5 mins and every 8 hours during confinement 
c.) Supine for 5 mins at Screening and at 3 hours after second dose on Day 7 
d.)  see Table 3 
 


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Table 3 
Schedule of PK Sampling for MAD 
 
PK Sampling (4 mL)  Study Day BID Dosing 
Time (hr) 
1 
2 
3 
4 
5 
6 
7 
8 
Pre-dose 
Xa 
Xa 
  
Xa 
  
Xa 
Xa 
Xa 
0.5 
X 
  
  
  
  
  
X 
  
1 
X 
  
  
  
  
  
X 
  
2 
X 
  
  
  
  
  
X 
  
4 
X 
  
  
  
  
  
X 
  
6 
X 
  
  
  
  
  
X 
  
8 
X 
  
  
  
  
  
X 
  
10 
X 
  
  
  
  
  
X 
  
12 
X 
Xa 
  
Xa 
  
Xa 
Xa 
  
 
 
 
 
 
 
 
 
 
  
PK Sampling  (4 mL)  Study Day TID Dosing 
Time (hr) 
1 
2 
3 
4 
5 
6 
7 
8 
Pre-dose 
Xa 
Xa 
  
Xa 
  
Xa 
Xa 
Xa 
0.5 
X 
  
  
  
  
  
X 
  
1 
X 
  
  
  
  
  
X 
  
2 
X 
  
  
  
  
  
X 
  
4 
X 
  
  
  
  
  
X 
  
6 
X 
  
  
  
  
  
X 
  
8 
X 
Xa 
  
Xa 
  
Xa 
Xa 
  
a Pre-dose 


CONFIDENTIAL 
 
ANA Therapeutics 
Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
3. 
Introduction 
Niclosamide is a chlorinated salicylanilide with anthelmintic, antiviral and anti-inflammatory 
activity being developed as a potential treatment for COVID-19. Niclosamide was discovered in 
1958, and was approved by the Food and Drug Administration (FDA) in 1982 under New Drug 
Application (NDA) 018669 (Bayer Pharmaceuticals) for the treatment of tapeworm infections, 
although it has since been voluntarily discontinued from marketing in the United States by Bayer. 
Niclosamide is approved in several other countries for the treatment of tapeworm infections and is 
on the World Health Organization's List of Essential Medicines. Due to its pleiotropic 
pharmacologic activity, niclosamide has been and continues to be studied for a variety of other 
clinical indications.  
ANA Therapeutics is developing ANA001, an orally administered niclosamide formulation 
containing 250 mg niclosamide per capsule, to treat adult patients with coronavirus disease of 2019 
(COVID-19). COVID-19 results in a cytokine release syndrome (Hirano and Murakami, 2020) 
that is associated – amongst others - with acute respiratory distress syndrome (ARDS), a 
widespread inflammation in the lungs, and increased blood clotting. Niclosamide is a promising 
oral therapeutic which has both antiviral and anti-inflammatory properties and a long history of 
safety in humans.   
Niclosamide has broad in vitro antiviral activity (Fan et al., 2019, Andersen et al., 2019, Jurgeit et 
al., 2012, Mazzon et al., 2019, Xu et al., 2020, Wu et al., 2004, Wen et al., 2007, Gassen et al., 
2019) including activity against severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) 
with a half-maximal inhibitory concentration (IC50) of 0.15-0.28 µM (49-92 ng/mL) (Shi, 2020, 
unpublished data, Jeon et al., 2020, Gassen et al., 2020). Additionally, niclosamide has in vitro 
anti-inflammatory properties due to its inhibition of nuclear factor kappa-light-chain enhancer of 
activated B cells (NF-κB) and signal transducer and activator of transcription 3 (STAT3), two 
main drivers to induce the expression of proinflammatory cytokines. NF-κB and STAT3 were 
inhibited by niclosamide with an IC50 of 0.125 µM (40.9 ng/mL) (Jin et al., 2010) and 0.25 µM 
(81.8 ng/mL), respectively (Ren et al., 2010). In addition, a recent in vitro study showed that 
niclosamide works as a potent bronchodilator that relaxed histamine induced constriction of mouse 
tracheal rings and human bronchial rings with a half-maximal effective concentration (EC50) of 
124 ng/mL (0.4 µM) and 240 ng/mL (0.7 µM), respectively (Miner et al., 2019). 
PK data available in the literature show significant variability. A single dose of 2000 mg reaches 
maximal systemic serum concentrations in humans of 0.76 μM to 18.3 μM (249 to 5986 ng/mL) 
(Andrews et al., 1982). A study with prostate cancer patients showed that 149-182 ng/mL (0.46-
0.56 µM) become available after a single oral dose of 1000 mg (Schweizer et al., 2018). These 
studies strongly suggest that the dose regimen foreseen for clinical development – between 1000 
mg BID, 1000 mg TID and 2000 mg BID - will provide sufficient systemic and intracellular drug 
levels for effective antiviral and anti-inflammatory activity. However, there is a need for well 
characterized PK data to support dose selection in COVID-19 and other indications. 
 


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ANA Therapeutics 
Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
As niclosamide was approved by FDA to treat tapeworms in humans, it has a well-understood 
clinical safety profile on oral administration at 2000 mg daily for up to 7 days. In addition, it has 
been studied in clinical trials in oncology and several other indications, and two other Sponsors 
have recently initiated trials of niclosamide in COVID-19 patients. This Investigational New Drug 
Application represents the first clinical use of ANA001 capsules, thus there is no previous clinical 
data to report for this formulation of niclosamide. 
3.1. 
Study Rationale 
ANA001 is a new formulation containing 250 mg niclosamide per capsule. Although other 
niclosamide formulations have been studied for tapeworm infections and other potential human 
uses, limited systemic PK/PD data exists in the literature. To ensure proper systemic levels of 
niclosamide are achieved for therapeutic effect, a SAD/MAD study with ANA001 capsules is 
necessary. 
3.2. 
Background 
Niclosamide has broad in vitro antiviral activity (Fan et al., 2019, Andersen et al., 2019, Jurgeit et 
al., 2012, Mazzon et al., 2019, Xu et al., 2020, Wu et al., 2004, Wen et al., 2007, Gassen et al., 
2019) including activity against coronaviruses (Figure 1). Three initial publications have 
demonstrated that niclosamide inhibited replication of both SARS-CoV at concentrations of 
≥1.56 µM (≥510 ng/mL) (Wu et al., 2004) and 40 µM (13,084 ng/mL) (Wen et al., 2007) and 
Middle East respiratory syndrome (MERS)-CoV with an IC50 of 0.32 µM (105 ng/mL) (Gassen et 
al., 2019).   
Figure 1 
Phylogenetic Tree Showing Antiviral Effects of Niclosamide Against Various 
Virus Families 
 
Source: Adapted from Xu et al 2020. 


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ANA Therapeutics 
Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
SARS-CoV, MERS-CoV and SARS-CoV-2 share 86% homology (Wilder-Smith et al., 2020) and 
three recent studies have confirmed the in vitro antiviral efficacy of niclosamide against 
SARSCoV-2. The reported IC50 values were 0.15 µM (49 ng/mL) (Shi, 2020, unpublished data), 
0.28 µM (92 ng/mL) (Jeon et al., 2020) and 0.17 µM (56 ng/mL) (Gassen et al., 2020). 
Furthermore, it was shown that pretreating cells for 24 h with 5 µM (1636 ng/mL) niclosamide 
followed by drug washout and viral infection reduced SARS-CoV-2 replication significantly 
(Gassen et al., 2020). This indicates that niclosamide has potential as a prophylactic COVID-19 
treatment and that serum concentrations may not need to continuously exceed in vitro inhibitory 
levels to be effective. 
3.2.1. 
Chemical Name/Structure 
USAN:  Niclosamide 
IUPAC:  5-Chloro-N-(2-chloro-4- nitrophenyl)-2-hydroxybenzamid 
Molecular formula: C13H8Cl2N2O4 
Molecular weight: 327.1 g/mol 
 
3.2.2. 
Nonclinical Studies 
Nonclinical Pharmacology 
In an in vitro study, Vero E6 cells were infected with a SARS-CoV-2 reporter virus. This virus has 
a luciferase gene engineered into its genome and when cells are infected with such a reporter virus, 
they express luciferase that can be quantified to measure viral replication. Increasing 
concentrations of niclosamide were added to Vero E6 cells together with reporter SARS-CoV-2. 
Twenty-four hours post-infection, infected cells were assayed for luciferase activity. The antiviral 
IC50 of niclosamide was 0.15 µM (49 ng/mL) (Shi, 2020, unpublished data). 
In a separate in vitro study, 50 FDA-approved drugs were screened for their ability to inhibit 
SARS-CoV-2 replication. Vero cells were infected with SARS-CoV-2 in the absence and presence 
of various drug concentrations. 24 hours post infection, cells were scored by immunofluorescence 
analysis with an antibody specific for the viral N protein of SARS-CoV-2 to generate dose-
response curves for each compound tested. Niclosamide demonstrated an IC50 of 0.28 µM 
(92 ng/mL). In the same assay, the IC50 values of remdesivir, chloroquine and lopinavir were 
11.41 µM (3732 ng/mL), 7.28 µM (2381 ng/mL) and 9.12 µM (2983 ng/mL), respectively (Jeon 
et al., 2020).  
In an additional in vitro study, VeroFM cells were infected with SARS-CoV-2 and treated with 
increasing concentrations of niclosamide. 48 hours post infection, plaque forming units were 
determined by plaque assay. The antiviral IC50 of niclosamide was 0.17 µM (56 ng/mL). 


CONFIDENTIAL 
 
ANA Therapeutics 
Protocol ANA001 
Page 23 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
Furthermore, pretreating VeroFM cells for 24 h with 5 µM niclosamide significantly reduced viral 
replication (Gassen et al., 2020). 
Nonclinical Toxicology 
Oral toxicology studies for NDA 018669 evaluated niclosamide in mice, rats, rabbits, dogs, and 
cats. Single dose studies included doses as high as 5000 mg/kg in rats without any lethality 
(reviewed in Andrews et al., 1982). A repeat dose study with dogs showed that single oral doses 
of 6000 mg/kg for 8 consecutive days, followed by 4500 mg/kg for another 24 consecutive days 
were tolerated well with no signs of intoxication (reviewed in Andrews et al., 1982). Other repeat 
dose studies in rabbits, cats or dogs with doses that were 2-5 times the human therapeutic dose and 
lasted 11, 12, and 84-96 days were non toxic, based on routine laboratory tests and toxicology. 
Male cats were noted to have a mild reduction in white blood cells (WBC), and dogs showed a 
tendency toward watery stools, but otherwise adverse treatment effects were not evident. Other 
studies reported that doses of 250 mg/kg may cause vomiting in dogs and cats. The conclusion of 
these studies was that niclosamide was a low risk for dosing orally in humans at the standard 
therapeutic regimen of 2000 mg/day (BAYER NDA 018669; Summary Basis for Approval 
(SBA)). 
3.2.3. 
Safety 
Between 1971 and 1978, niclosamide was administered to 6365 patients under a US IND. Doses 
were up to and including 2000 mg/day for 7 days. There were 2385 evaluable patients, of which 
13.3% reported side effects, all of which were mild or moderate, with none requiring treatment 
discontinuation. These included nausea/emesis in 4.1%, abdominal discomfort/loss of appetite in 
3.4%, diarrhea in 1.6%, drowsiness/dizziness/headache in 1.4%, and skin rash/pruritis in 0.3% of 
patients (NDA 018669 Review Documentation).  
In addition, according to the Safety Review of Niclosamide, Pyrantel, Triclabendazole and 
Oxamniquine report of the World Health Organization (WHO), there have been 84 reported 
adverse drug reactions related to niclosamide between 1975 and 2004 in the WHO database. They 
include 173 reports from 16 countries and the most common reactions are related to skin and 
appendages (41 reports), gastrointestinal tract (37 reports), cardiovascular system (28 reports) and 
anaphylactic reactions (9 reports). 
Niclosamide was approved as a pregnancy class B drug and is thus safe to use during pregnancy. 
Summarized below are safety assessment findings reported in the available literature:  
• Prostate cancer patients received 500 mg and 1000 mg of niclosamide orally thrice per day 
for 15 days. TID regimens of >500 mg PO per day suggested the potential for DLT, 
however these findings are confounded by coadministration of antineoplastic treatment 
associated with a similar range of GI disturbances and occurred after niclosamide was 
dosed for >7 days (Schweizer et al., 2018). 
• Colorectal cancer patients received 2000 mg of niclosamide orally once per day until 
disease progression or toxicity (up to four months). No adverse side effects were reported 
(Burock et al., 2020). 


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ANA Therapeutics 
Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
• 2000 mg niclosamide/person did not result in hematological changes (Andrews et al., 
1982). 
• No induction of methemoglobinemia was observed in men after oral administration of 30 
mg/kg b.w. (~2000 mg/person) (Andrews et al., 1982). 
• Men and women received 1000 mg niclosamide once or twice per person; children (6-15 
years of age) received 750-1000 mg/person; neither scenario resulted in signs of 
intoxication (Andrews et al., 1982). 
• In a Phase I study, adults were divided into two groups (niclosamide and placebo). Study 
participants received 2000 mg/day for 3 days with a repeat course 6 days later. Transitory 
reactions, such as loose stools and stomach aches were reported (Andrews et al., 1982). 
• No dermal sensitizing effect was observed upon topical administration (Andrews et al., 
1982). 
At the established doses, most adverse events were transitory and generally mild to moderate.  
3.3. 
Benefit/Risk Assessment 
Niclosamide is approved for dosing up to 2000 mg PO daily for 7 days. There is limited safety and 
PK data in humans receiving more than 2000 mg as a single dose and/or more frequently than once 
daily. The current SAD/MAD study is designed to explore the safety and PK of dosing up to 3000 
mg/day as a single dose and regimens not to exceed 2000 mg/day as divided doses BID or TID. In 
light of the immediate pressing need for development of an effective and safe oral therapeutic that 
can prevent progression of COVID-19, the benefit to risk ratio for the study is anticipated to be 
justified. 
More detailed information about the known and expected benefits and risks and reasonably 
expected adverse events of ANA001 may be found in the Investigator’s Brochure. 
 


CONFIDENTIAL 
 
ANA Therapeutics 
Protocol ANA001 
Page 25 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
4. 
Objectives and Endpoints 
Objectives 
Endpoints 
Primary 
 
• To assess safety and tolerability of 
single oral daily doses of ANA001 
(1000 mg, 2000 mg and 3000 mg) 
• To assess safety and tolerability of 
multiple daily oral doses (twice daily 
[BID] or thrice daily [TID]) of 
ANA001 for 7 days 
• Incidence of adverse events (AEs), 
serious AEs (SAEs), study drug 
discontinuation due to an AE (MAD 
portion only), use of concomitant 
medications, and change from baseline 
in clinical laboratory tests, vital signs, 
electrocardiograms (ECG), and 
physical examinations 
Secondary 
 
• To assess the PK of single and 
multiple doses of ANA001 
• AUC0-t, AUC0-last, AUC0-∞ (SAD 
only), Cmax, tmax, t1/2, CL/F, Vz/F 
 
 


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ANA Therapeutics 
Protocol ANA001 
Page 26 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
5. 
Study Design 
5.1. 
Overall Design 
This is a randomized, double-blind study to be conducted in two parts: single ascending dose 
(SAD) and multiple ascending dose (MAD). Potential participants for each part will undergo 
screening procedures within 30 days of enrolment.  
SAD 
On Day -1, eligible participants will be randomly assigned to active drug or placebo in an 8:2 ratio. 
On Day 1, following an overnight fast of at least 10 hours, participants will receive their assigned 
treatment (1000 mg, 2000 mg, or 3000 mg of ANA001) with a standardized light meal (500 to 
750 cal).  A baseline ECG will be performed at Screening and at 3 hours after dosing. Blood 
pressure and heart rate will be measured every 8 hours while confined to the clinic. Blood for PK 
will be collected before dosing and at 0.5, 1, 2, 4, 6, 8, 10, 12 and 24 hours after dosing. Participants 
will be discharged on Day 2 following the 24-hour PK sample and completion of clinical 
laboratory tests and physical examination. They will return on Day 7 (±2) for follow-up procedures 
including physical examination, laboratory tests, and assessment of AEs and concomitant 
medication use. 
MAD 
On Day -1, eligible participants will be randomly assigned to active drug or placebo in a 9:3 ratio 
on a BID (q12h) or TID (q8h) schedule, to be determined based on the PK results of the SAD 
cohorts. Total daily doses will not exceed 2000 mg of ANA001. On Days 1 through 7, participants 
will receive their assigned treatment in the morning with a standardized light meal (500 to 750 cal) 
following an overnight fast of at least 10 hours. Afternoon or evening doses will be given with a 
light snack.  A baseline ECG will be performed at Screening and at 3 hours after the second dose 
on Day 7. Blood pressure and heart rate will be measured every 8 hours while confined to the 
clinic.  
During BID dosing, blood for PK will be collected on Days 1 and 7 before dosing  and at 0.5, 1, 
2, 4, 6, 8, 10, and 12 hours after dosing. Blood samples will also be collected on Days 2, 4, 6, and 
8 before dosing and before the 12 hour dose on Days 2, 4, and 6.  
During TID dosing, blood for PK will be collected on Days 1 and 7 before dosing and at 0.5, 1, 2, 
4, 6, and 8 hours after dosing. Blood samples will also be collected on Days 2, 4, 6, and 8 before 
dosing, before the 8 hour dose on Days 2, 4, and 6, and before the 16 hour dose on Day 6. 
Participants will be discharged on Day 8 following the last PK sample and completion of clinical 
laboratory tests and physical examination. They will return on Day 15 (±2) for follow-up 
procedures including physical examination, laboratory tests, and assessment of AEs and 
concomitant medication use. 


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ANA Therapeutics 
Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
5.2. 
Participant and Study Completion 
A minimum of 30 participants will be enrolled in the SAD part and up to 36 participants in the 
MAD part. At least 3 participants of each sex will be included in each cohort. 
5.3. 
End of Study Definition 
A participant is considered to have completed the study if he/she has completed all phases of the 
study including the last scheduled procedure shown in the Schedule of Activities. 
The end of the study is defined as the date of the last visit of the last participant in the study. 
5.4. 
Scientific Rationale for Study Design 
Sequential timing of the ascending dose cohorts will allow early stopping if serious or 
unanticipated safety signals are observed at any dose level. 
5.5. 
Justification for Dose 
Niclosamide was approved by FDA to treat tapeworms in humans and has a well understood 
clinical safety profile with oral administration at 2000 mg daily for up to 7 days. Furthermore, a 
single dose of 2000 mg reaches maximal systemic serum concentrations in humans of 0.76 μM to 
18.3 μM (249 ng/mL to 5986 ng/mL) (Andrews et al., 1982). A study with prostate cancer patients 
showed that 149 ng/mL to 182 ng/mL (0.46 µM to 0.56 µM) become available after a single oral 
dose of 1000 mg (Schweizer et al., 2018). In a very recent study, colorectal cancer patients received 
2000 mg of niclosamide orally once a day until disease progression or toxicity (up to four months). 
Plasma levels mainly peaked 240 minutes after the first niclosamide administration with a median 
Cmax of 2.03 µM (665 ng/mL) (Burock et al., 2020). The lower bounds of the reported Cmax values 
fall within the effective in vitro ranges of niclosamide as an antiviral agent (0.15-0.28 µM, 49-92 
ng/mL) (Shi, 2020, unpublished results; Jeon et al., 2020, Gassen et al., 2020) and an anti-
inflammatory drug that inhibits NF-kB and STAT3 at 0.13 µM (42.5 ng/mL) (Jin et al., 2010) and 
0.25 µM (81.8 ng/mL) (Ren et al., 2010), respectively. 
These studies strongly suggest that the dose regimens between 1000 mg BID to 2000 mg BID or 
1000 mg TID to 1500 mg TID will provide sufficient systemic and intracellular drug levels for 
effective antiviral and anti-inflammatory activity. The current study is designed to assess the safety 
of single doses of niclosamide up to 3000 mg and multiple dosing regimens not to exceed 2000 
mg daily divided BID or TID. The PK data from this study will be used to refine the optimum dose 
and frequency of administration to achieve antiviral and anti-inflammatory activity in humans. 
 


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ANA Therapeutics 
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Page 28 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
6. 
Study Population 
Prospective approval of protocol deviations to recruitment and enrollment criteria, also known as 
protocol waivers or exemptions, is not permitted. 
6.1. 
Inclusion Criteria 
Participants are eligible to be included in the study only if all of the following criteria apply: 
1. Signed the COVID and study informed consent forms as described in Appendix 3 which 
includes compliance with the requirements and restrictions listed in the informed consent 
form (ICF) and in this protocol 
2. Man or woman, 18 to 65 years of age inclusive at the time of signing the informed 
consent form 
3. Overtly healthy as determined by medical evaluation including medical history, physical 
examination, clinical laboratory tests 
4. Body mass index (BMI) within 18 to 30.0 kg/m2 (inclusive) and body weight not less 
than 50 kg 
5. Blood pressure (after supine for 5 minutes) at Screening and Day -1 between 90 and 140 
mmHg systolic, inclusive, and no higher than 90 mmHg diastolic. 
6. A 12-lead electrocardiogram (ECG) at Screening consistent with normal cardiac 
conduction and function, including: 
• Sinus rhythm 
• Pulse rate between 50 and 100 beats per minute (bpm) 
• QTc interval  450 milliseconds (QT interval corrected using Fridericia correction 
method [QTcF]) 
• QRS interval of <120 milliseconds 
• PR interval <200 milliseconds 
• Morphology consistent with healthy cardiac conduction and function 
7. Non-smoker or ex-smoker for >12 months 
8. If male, must agree to use contraception as detailed in Appendix 5 of this protocol during 
the treatment period and for at least 30 days (a spermatogenesis cycle) after the last dose 
of study treatment and refrain from donating sperm during this period 
9. If female, is not pregnant (see Appendix 5), not breastfeeding, and meets at least one of 
the following conditions: 
Not a woman of childbearing potential (WOCBP) as defined in Appendix 5 
OR 


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Status: Final v2.0; Date 09Jan2021 
 
A WOCBP who agrees to follow the contraceptive guidance in Appendix 5 during 
the treatment period and for at least 30 days (one menstrual cycle) after the last 
dose of study treatment. 
Note: Retesting of abnormal laboratory values that may lead to exclusion will be allowed once. 
Retesting will take place during an unscheduled visit in the screening phase. 
6.2. 
Exclusion Criteria  
Participants are excluded from the study if any of the following criteria apply: 
1. Has a history of or current clinically significant medical illness including but not limited 
to: cardiac arrhythmias or other cardiac disease; hematologic disease; coagulation 
disorders (including any abnormal bleeding or blood dyscrasias); lipid abnormalities; 
significant pulmonary disease, including bronchospastic respiratory disease; diabetes 
mellitus; hepatic or renal insufficiency (creatinine clearance below 60 mL/min); thyroid 
disease; neurologic or psychiatric disease; infection; or any other illness that the 
Investigator considers should exclude the subject or that could interfere with the 
interpretation of the study results. 
2. Has known allergy to niclosamide or salicylate-containing medications. 
3. Clinically significant abnormal values for hematology, clinical chemistry, or urinalysis at 
screening as assessed per Appendix 6 
4. Clinically significant abnormal physical examination, vital signs or 12 lead ECG at 
screening as deemed appropriate by the Investigator 
5. Has a history of human immunodeficiency virus (HIV) antibody positive, or tests positive 
for HIV; has a history of hepatitis B surface antigen (HBsAg) or hepatitis C antibody 
(anti-HCV) positive, or other clinically active liver disease, or tests positive for HBsAg 
or anti-HCV at Screening. 
6. History of drug or alcohol abuse according to Diagnostic and Statistical Manual of 
Mental Disorders (4th edition) criteria within 5 years before screening or positive test 
result(s) for alcohol and/or drugs of abuse at screening and admission 
7. Has received an investigational drug or used an invasive investigational medical device 
within 1 month or within a period less than 10 times the drug’s half-life, whichever is 
longer, before Day 1. 
8. Has preplanned surgery or procedures that would interfere with the conduct of the study 
9. Is an employee of the Investigator or study site, with direct involvement in the proposed 
study or other studies under the direction of that Investigator or study site, as well as 
family members of the employees or the Investigator 


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ANA Therapeutics 
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6.3. 
Lifestyle Restrictions 
6.3.1. 
Meals and Dietary Restrictions 
1. Participants will refrain from consumption of red wine, Seville oranges, grapefruit or 
grapefruit juice, pomelos, exotic citrus fruits, grapefruit hybrids, or fruit juices from 3 
days before the start of study treatment until after the final dose. 
6.3.2. 
Caffeine, Alcohol, and Tobacco 
1. Participants will abstain from ingesting caffeine- or xanthine-containing products (e.g., 
coffee, tea, cola drinks, and chocolate) for 24 hours before the start of dosing until after 
collection of the final pharmacokinetic (PK) blood sample. 
2. Participants will abstain from alcohol for 24 hours before the start of dosing until after 
collection of the final PK blood sample.  
3. Use of tobacco products will not be allowed during the study. 
6.3.3. 
Activity 
1. Participants will abstain from strenuous exercise for 24 hours before each blood collection 
for clinical laboratory tests. Participants may participate in light recreational activities 
during studies (e.g., watching television, reading). 
6.4. 
Screen Failures 
Screen failures are defined as participants who consent to participate in the clinical study but are 
not subsequently entered into the study. A minimal set of screen failure information is required 
to ensure transparent reporting of screen failure participants to meet the Consolidated Standards 
of Reporting Trials (CONSORT) publishing requirements and to respond to queries from 
regulatory authorities. Minimal information includes demography, screen failure details, 
eligibility criteria, and any serious adverse event (SAE). 
Individuals who do not meet the criteria for participation in this study (screen failure) may not be 
rescreened. 


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7. 
Treatments 
Study treatment is defined as any investigational treatment(s), marketed product(s), placebo, or 
medical device(s) intended to be administered to a study participant according to the study 
protocol. 
7.1. 
Treatments Administered 
Study Treatment Name: 
ANA001 
Placebo 
Dosage formulation: 
Size 0 capsule 
Size 0 capsule 
Unit dose strength(s)/ 
Dosage level(s): 
250 mg/capsule 
NA 
Route of Administration  
oral 
oral 
Dosing instructions: 
 
4, 6, 8, or 12 capsules 
with light meal or snacka 4, 6, 8, or 12 capsules 
with light meal or snacka 
Packaging and Labeling 
Bulk bottle  
Storage conditions 
Capsules (ANA001 or matching placebo) should be 
stored at controlled room temperature (20-25 °C) out 
of direct sunlight. 
Manufacturer 
ANA Therapeutics, Inc. 
a 500 to 750 cal 
7.2. 
Dose Modification 
The dose(s) and schedule (BID or TID) in the MAD part will be determined following review of 
the safety and PK data from the SAD part. Total daily doses will not exceed 2000 mg. 
7.3. 
Method of Treatment Assignment 
Within each cohort, participants will be randomly assigned to active drug or placebo as they qualify 
for the study using a computer-generated randomization. 
7.4. 
Blinding 
Participants will be randomly assigned to receive study treatment. Investigators will remain 
blinded to each participant’s assigned study treatment throughout the course of the study. In order 
to maintain this blind, an otherwise uninvolved 3rd party (e.g., site pharmacist) will be responsible 
for the dispensation of all study treatment and will endeavor to ensure that there are no differences 
in time taken to dispense following randomization. 


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7.5. 
Preparation/Handling/Storage/Accountability 
The Investigator or designee must confirm appropriate temperature conditions have been 
maintained during transit for all study treatment received and any discrepancies are reported and 
resolved before use of the study treatment. 
Only participants enrolled in the study may receive study treatment and only authorized site staff 
may supply or administer study treatment. All study treatments must be stored in a secure, 
environmentally controlled, and monitored (manual or automated) area in accordance with the 
labeled storage conditions with access limited to the Investigator and authorized site staff. 
The Investigator, institution, or the head of the medical institution (where applicable) is responsible 
for study treatment accountability, reconciliation, and record maintenance (i.e., receipt, 
reconciliation, and final disposition records). 
7.6. 
Treatment Compliance 
Participants will receive study treatment under the direct supervision of the Investigator or 
designee. A hand- and mouth check will be done to ensure that the participants swallowed the 
drugs. 
7.7. 
Concomitant Therapy 
Any medication or vaccine (including over-the-counter or prescription medicines, vitamins, 
and/or herbal supplements) that the participant is receiving at the time of enrollment or receives 
during the study must be recorded along with: 
• Reason for use 
• Dates of administration including start and end dates 
• Dosage information including dose and frequency 
The Medical Monitor should be contacted if there are any questions regarding concomitant or 
prior therapy. 
Participants must abstain from taking prescription or nonprescription drugs (including vitamins 
and dietary or herbal supplements) within 7 days (or 14 days if the drug is a potential enzyme 
inducer) or 5 half-lives (whichever is longer) before the start of study treatment until completion 
of the follow-up visit, unless, in the opinion of the Investigator and Sponsor, the medication will 
not interfere with the study.  
Acetaminophen, at doses of  2 grams/day, is permitted for use any time during the study. Other 
concomitant medication may be considered on a case-by-case basis by the Investigator. 


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7.7.1. 
Rescue Medicine 
No rescue medication will be provided.  
7.8. 
Treatment after the End of the Study 
Participants will not receive study drug (ANA001 or matching placebo) after the end of the 
study. 


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8. 
Discontinuation/Withdrawal Criteria  
8.1. 
Discontinuation of Study Treatment  
An unexpected AE related to study drug could lead to temporary or permanent discontinuation of 
a participant’s study treatment, as determined by the Medical Monitor or Investigator.   
8.2. 
Withdrawal from the Study 
A participant may withdraw from the study at any time at his/her own request, or may be 
withdrawn at any time at the discretion of the Investigator for safety, behavioral, compliance, or 
administrative reasons. 
If the participant withdraws consent for disclosure of future information, the Sponsor may retain 
and continue to use any data collected before such a withdrawal of consent. 
If a participant withdraws from the study, he/she may request destruction of any samples taken 
and not tested, and the Investigator must document this in the site study records. 
See SoA (Section 2) for data to be collected at the time of study discontinuation and follow-up 
and for any further evaluations that need to be completed. 
Participants who withdraw will not be replaced. 
8.3. 
Lost to Follow Up 
A participant will be considered lost to follow-up if he or she repeatedly fails to return for 
scheduled visits and is unable to be contacted by the study site.  
The following actions must be taken if a participant fails to return to the clinic for a required 
study visit: 
• The site must attempt to contact the participant and reschedule the missed visit as soon as 
possible and counsel the participant on the importance of maintaining the assigned visit 
schedule and ascertain whether or not the participant wishes to and/or should continue in 
the study. 
• Before a participant is deemed lost to follow up, the Investigator or designee must make 
every effort to regain contact with the participant (where possible, 3 telephone calls and, 
if necessary, a certified letter to the participant’s last known mailing address or local 
equivalent methods). These contact attempts should be documented in the participant’s 
medical record. 
• Should the participant continue to be unreachable, he/she will be considered to have 
withdrawn from the study with a primary reason of lost to follow-up.  


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9. 
Study Assessments and Procedures 
• Study procedures and their timing are summarized in the SoA. Protocol waivers or 
exemptions are not allowed. 
• Immediate safety concerns should be discussed with the Sponsor immediately upon 
occurrence or awareness to determine if the participant should continue or discontinue study 
treatment. 
• Adherence to the study design requirements, including those specified in the SoA, is essential 
and required for study conduct. 
• All screening evaluations must be completed and reviewed to confirm that potential 
participants meet all eligibility criteria. The Investigator will maintain a screening log to 
record details of all participants screened and to confirm eligibility or record reasons for 
screening failure, as applicable.  
• Procedures conducted as part of the participant’s routine clinical management (e.g., blood 
count) and obtained before signing of the ICF may be utilized for screening or baseline 
purposes provided the procedures met the protocol-specified criteria and were performed 
within the time frame defined in the SoA. 
Volume of Blood to be Collected From Each Subject - SAD 
Type of Sample 
Volume per Sample 
(mL) 
No. of Samples/ 
Subject 
Total Volume 
of Blood 
(mL)a 
Safety (including screening 
and post treatment 
assessments) 
 
 
 
- Hematology 
4 
4 
16 
- Serum chemistry 
8.5 
4 
34 
serology (HIV, hepatitis) 
8.5 
1 
8.5 
serum -hCG pregnancy 
test 
3 
1 
3 
Pharmacokinetic samples - 
SAD 
4 
10 
40 
Total – SAD 
 
 
101.5 
a Calculated as number of samples multiplied by amount of blood per sample. 
 
 


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Volume of Blood to be Collected From Each Subject - MAD 
Type of Sample 
Volume per Sample 
(mL) 
No. of Samples/ 
Subject 
Total Volume 
of Blood 
(mL)a 
Safety (including screening 
and post treatment 
assessments) 
 
 
 
- Hematology 
4 
6 
64 
- Serum chemistry 
8.5 
6 
51 
serology (HIV, hepatitis) 
8.5 
1 
8.5 
serum -hCG pregnancy 
test 
3 
1 
3 
Pharmacokinetic samples – 
MAD (BID/TID) 
4 
25/28 
100/112 
Total – BID 
Total - TID 
 
 
226.5 
238.5 
a Calculated as number of samples multiplied by amount of blood per sample. 
Approximately 102 mL of blood will be collected during the SAD part. During the MAD, 
approximately 227 mL will be collected following BID dosing and 239 mL during TID dosing. 
9.1. 
Safety Assessments  
Safety assessments include incidence and frequency of AEs/SAEs, study drug discontinuation 
due to an AE (MAD portion only), use of concomitant medications, and changes from baseline 
in clinical laboratory tests, vital signs, electrocardiograms (ECG), and physical examinations. 
9.1.1. 
Physical Examinations 
A complete physical examination, conducted at screening, will include, at a minimum, 
assessments of the Skin, Cardiovascular, Respiratory, Gastrointestinal, and Neurological 
systems. Height and weight will also be measured and recorded. 
A brief physical examination will include, at a minimum, assessments of the Cardiovascular, 
Respiratory, Gastrointestinal systems. 


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9.1.2. 
Vital Signs 
Vital signs should be taken in a quiet setting without distractions (e.g., television, cell phones) 
and before any blood collections. 
Oral temperature, pulse rate, respiratory rate, and blood pressure will be assessed. 
Blood pressure and pulse measurements will be assessed with a completely automated device 
after 3 to 5 minutes rest in a supine position. Manual techniques will be used only if an 
automated device is not available. 
Blood pressure and pulse measurements will be measured after 3 to 5 minutes rest in a supine 
position. 
9.1.3. 
Electrocardiograms 
A single ECG will be obtained for screening and at 3 hours after dosing during the SAD portion 
of the study and at Screening and 3 hours after dosing on Day 7 during the MAD portion. An ECG 
machine that automatically calculates the heart rate and measures PR, QRS, QT, and QTc intervals 
will be used. 
9.1.4. 
Clinical Safety Laboratory Assessments 
See Appendix 2 for the list of clinical laboratory tests to be performed and the SoA (Section 2) for 
the timing and frequency.  
The Investigator must review the laboratory report, document this review, and record any clinically 
relevant changes (as per the reference laboratory values [Appendix 6]) occurring during the study 
in the AE section of the CRF. The laboratory reports must be filed with the source documents. 
Clinically significant abnormal laboratory findings are those which are not associated with the 
underlying disease, unless judged by the Investigator to be more severe than expected for the 
participant's condition. 
All laboratory tests with values considered clinically significantly abnormal during participation 
in the study should be repeated until the values return to normal or baseline or are no longer 
considered clinically significant as per the reference laboratory values in Appendix 6. 
• If such values do not return to normal/baseline within a period of time judged reasonable 
by the Investigator, the etiology should be identified and the Sponsor notified. 
• All protocol-required laboratory assessments, as defined in Appendix 2, must be 
conducted in accordance with the laboratory manual and the SoA.  
• If laboratory values from non-protocol specified laboratory assessments performed at the 
institution’s local laboratory require a change in participant management or are considered 
clinically significant as given in Appendix 6 (e.g., SAE or AE or dose modification), then 
the results must be recorded in the CRF.  


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9.2. 
Adverse Events 
The definitions of an AE or SAE can be found in Appendix 4. 
AE will be reported by the participant (or, when appropriate, by a caregiver, surrogate, or the 
participant's legally authorized representative).  
The Investigator and any designees are responsible for detecting, documenting, and recording 
events that meet the definition of an AE or SAE and remain responsible for following up AEs that 
are serious, considered related to the study treatment or study procedures, or that caused the 
participant to discontinue the study.  
9.2.1. 
Time Period and Frequency for Collecting AE and SAE Information  
All AEs and SAEs will be collected from the signing of the ICF until the follow-up visit at the 
time points specified in the SoA (Section 2). 
Medical occurrences that begin before the start of study treatment but after obtaining informed 
consent will be recorded on the Medical History/Current Medical Conditions section of the case 
report form (CRF) not the AE section. 
All SAEs will be recorded and reported to the Sponsor or designee within 24 hours, as indicated 
in Appendix 4. The Investigator will submit any updated SAE data to the Sponsor within 24 hours 
of it being available. 
Investigators are not obligated to actively seek AEs or SAEs in former study participants. However, 
if the Investigator learns of any SAE, including a death, at any time after a participant has been 
discharged from the study, and he/she considers the event to be reasonably related to the study 
treatment or study participation, the Investigator must promptly notify the Sponsor. 
The method of recording, evaluating, and assessing causality of AEs and SAEs and the procedures 
for completing and transmitting SAE reports are provided in Appendix 4. 
9.2.2. 
Method of Detecting AEs and SAEs 
Care will be taken not to introduce bias when detecting AEs and/or SAEs. Open-ended and 
non-leading verbal questioning of the participant is the preferred method to inquire about 
AE occurrences.  
9.2.3. 
Follow-up of AEs and SAEs 
After the initial AE/SAE report, the Investigator is required to proactively follow each participant 
at subsequent visits/contacts. All SAEs will be followed until resolution, stabilization, the event is 
otherwise explained, or the participant is lost to follow-up (as defined in Section 8.3). Further 
information on follow-up procedures is given in Appendix 4. 


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9.2.4. 
Regulatory Reporting Requirements for SAEs 
Prompt notification by the Investigator to the Sponsor of an SAE is essential so that legal 
obligations and ethical responsibilities towards the safety of participants and the safety of a study 
treatment under clinical investigation are met.  
The Sponsor has a legal responsibility to notify both the local regulatory authority and other 
regulatory agencies about the safety of a study treatment under clinical investigation. The Sponsor 
will comply with country-specific regulatory requirements relating to safety reporting to the 
regulatory authority, Institutional Review Boards (IRB)/Independent Ethics Committees (IEC), 
and Investigators. 
Investigator safety reports must be prepared for suspected unexpected serious adverse reactions 
(SUSAR) according to local regulatory requirements and Sponsor policy and forwarded to 
Investigators as necessary. 
An Investigator who receives an Investigator safety report describing a SAE or other specific safety 
information (e.g., summary or listing of SAEs) from the Sponsor will review and then file it along 
with the Investigator’s Brochure and will notify the IRB/IEC, if appropriate according to local 
requirements. 
9.2.5. 
Pregnancy 
Details of all pregnancies in female participants and, if indicated, female partners of male 
participants will be collected after the start of study treatment and until 30 days after the last dose. 
If a pregnancy is reported, the Investigator should inform the Sponsor within 24 hours of learning 
of the pregnancy and should follow the procedures outlined in Appendix 5. 
Abnormal pregnancy outcomes (e.g., spontaneous abortion, fetal death, stillbirth, congenital 
anomalies, ectopic pregnancy) are considered SAEs. 
9.3. 
Stopping Rules in Response to SAEs or Other Events 
Dose-limiting toxicity AEs will be defined as Grade ≥3, treatment-emergent, treatment-related 
laboratory abnormalities (Appendix 6) or AEs per the grading system by US Department of 
Health and Human Services (CTCAE, Version 5.0)The following Stopping Criteria will be used: 
Clinical Findings: 
1. Anaphylaxis or Grade ≥3 skin rash or pruritis 
2. Grade ≥3 nausea, emesis or diarrhea 


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Laboratory Findings: 
1. Evidence of new onset of acute renal or hepatic failure, defined by increases in 
BUN, creatinine, or LFTs (AST, ALT, Alkaline phosphatase) that reflect new 
evidence of Grade 3 thresholds for liver function tests, for creatinine, and for 
electrolytes attributable to the administration of drug in the context of the 
Investigator’s evaluation.  
2. Evidence of acute infection or inflammation, defined by changes in the WBC and 
differential, attributable to the administration of drug in the context of the 
Investigator’s evaluation.  
3. Any “unexpected” Grade ≥3 laboratory test result.  
If one of the above clinical or laboratory criteria for stopping occurs in any subject, further 
study recruitment and subject dosing will be halted until a full safety assessment can be 
completed. Unblinding of the affected subject will be performed and the PI and Sponsor will 
review the totality of clinical data and any other circumstances possibly related to the “Stopping 
Criteria” findings to determine if the study can be resumed, modified, or will be permanently 
terminated.  
9.4. 
Treatment of Overdose 
For this study, any dose of ANA001 greater than 3000 mg within a 24-hour time period will be 
considered an overdose. 
In the event of an overdose, the Investigator should: 
1. Contact the Medical Monitor immediately. 
2. Closely monitor the participant for any AE/SAE and laboratory abnormalities until 
ANA001 can no longer be detected systemically (at least 7 days). 
3. Document the quantity of the excess dose as well as the duration of the overdose in the 
CRF. 
Decisions regarding dose interruptions or modifications will be made by the Investigator in 
consultation with the Medical Monitor based on the clinical evaluation of the participant. 
9.5. 
Pharmacokinetics 
Whole blood samples of approximately 4 mL will be collected for measurement of plasma 
concentrations of ANA001 as specified in the SoA (Section 2). Additional samples may be 
collected during the study if warranted and agreed upon between the Investigator and the Sponsor. 


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Instructions for the collection and handling of biological samples will be provided by the Sponsor. 
The actual date and time (24-hour clock time) of each sample will be recorded. 
Any changes in the timing or addition of time points for any planned study assessments must be 
documented and approved by the relevant study team member and then archived in the Sponsor 
and site study files, but will not constitute a protocol amendment. The IRB/IEC will be informed 
of any safety issues that require alteration of the safety monitoring scheme or amendment of the 
ICF. 
The following PK parameters for ANA001 will be estimated using actual sample collection 
times: 
Cmax 
maximum plasma concentration during a dosing interval 
Cmin 
minimum plasma concentration during a dosing interval 
tmax 
time to reach the maximum plasma concentration 
AUCt 
area under the plasma concentration-time curve from time 0 to time t after dosing 
AUC0-last 
area under the plasma concentration-time curve from time 0 to time the last 
quantifiable concentration 
AUC0-∞ 
area under the plasma concentration-time curve from time 0 to infinity 
t1/2 
elimination half-life associated with the terminal slope (z) of the 
semilogarithmic drug concentration-time curve, calculated as 0.693/z 
CL/F 
Systemic clearance 
Vz/F 
Volume of distribution 
 
9.6. 
Medical Resource Utilization and Health Economics 
Not applicable. 


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10. 
Statistical Considerations  
10.1. 
Sample Size Determination  
Sample size was not based on statistical considerations. 
10.2. 
Populations for Analyses 
For purposes of analysis, the following populations are defined: 
Population 
Description 
Enrolled  
All participants who sign the ICF 
Safety 
All participants randomly assigned to study treatment and who 
take at least 1 dose of study treatment. Participants will be 
analyzed according to the treatment they actually received. 
Pharmacokinetic 
All participants randomly assigned to study treatment, who take 
at least 1 dose of study treatment, and contribute at least 1 post-
dose evaluable PK sample. Participants will be analyzed 
according to the dose they actually received. 
10.3. 
Statistical Analyses 
The statistical analysis plan will be developed and finalized before database lock and will describe 
the participant populations to be included in the analyses, and procedures for accounting for 
missing, unused, and spurious data. This section is a summary of the planned statistical analyses 
of the primary and secondary endpoints. 
10.3.1. Safety Analyses 
All safety analyses will be performed on the Safety Population. Safety parameters will include 
frequency of treatment-emergent AEs (TEAEs), SAEs, study drug discontinuation due to an AE 
(MAD portion only), concomitant medication use, and changes from baseline in clinical laboratory 
results, vital signs, ECG, and physical examinations. TEAEs are defined as AEs that started or 
worsened after dosing. Adverse events will be coded using the latest version of the Medical 
Dictionary for Regulatory Activities. A list of participants who have SAEs and who discontinue 
from the study due to an AE will be provided. Treatment-emergent AEs will be summarized by 
system organ class and preferred term for each dose level. 
Individual results of laboratory tests (chemistry, hematology, and urinalysis) will be listed and 
summarized. Baseline, post-dose, and change from baseline to post-dose laboratory data will be 
summarized by dose. Individual results of vital signs will be listed and summarized. Observed 
values and changes from baseline in vital signs will be summarized by dose. Individual results of 
ECG parameters from the 12-lead safety ECGs will be listed and changes from baseline will be 


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summarized by dose. Physical examination findings and concomitant medication use will be 
presented in data listings.  
10.3.2. Pharmacokinetic Analyses  
The non-compartmental PK analyses will be described in the Statistical Analysis Plan finalized 
before database lock. Plasma concentrations of ANA001 will be summarized by dose over each 
scheduled sampling time using descriptive statistics. Individual plasma concentration data versus 
actual time points will be presented in listings. Pharmacokinetic parameters of ANA001 will be 
summarized by dose using descriptive statistics. Individual PK parameters will be presented in a 
data listing. Dose proportionality will be assessed graphically and using the power model. 
10.3.3. Interim Analyses  
Data from the SAD Cohorts S1 through S3 will be summarized and evaluated to determine the 
doses and schedule to be used in a potential 4th SAD cohort and the BID and TID MAD cohorts. 
The Statistical Analysis Plan will describe the planned interim analyses in greater detail. 


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Espinosa-Aguirre, J. J. et al. Mutagenic activity of 2-chloro-4-nitroaniline and 5-chlorosalicylic 
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Lin, C. K. et al. Preclinical evaluation of a nanoformulated antihelminthic, niclosamide, in 
ovarian cancer. Oncotarget. (2016). doi: 10.18632/oncotarget.7113  
 
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anticancer activity of repurposed drug: Niclosamide. Carbohydr Polym. (2019). 
doi:10.1016/j.carbpol.2019.02.041  
 
Lu, D. et al. Metabolism of the anthelmintic drug niclosamide by cytochrome P450 enzymes and 
UDP-glucuronosyltransferases: metabolite elucidation and main contributions from CYP1A2 and 
UGT1A1. Xenobiotica. (2015). doi: 10.3109/00498254.2015.1047812 
 
Mazzon, M. et al. Identification of broad-spectrum antiviral compounds by targeting viral entry. 
Viruses (2019). doi:10.3390/v11020176  
 
Mehra, M. R. et al. Hydroxychloroquine or chloroquine with or without a macrolide for 
treatment of COVID-19: a multinational registry analysis. Lancet. (2020). doi: 
10.1016/S01406736(20)31180-6 
 
Miner, K. et al. Drug repurposing: The anthelmintics niclosamide and nitazoxanide are potent 
TMEM16A antagonists that fully bronchodilate airways. Front. Pharmacol. (2019) 
doi:10.3389/fphar.2019.00051  
 
Nair, A. B and S. A. Jacob. A simple practice guide for dose conversion between animals and 
human. J Basic Clin Pharma. (2016). doi: 10.4103/0976-0105.177703 NDA 018669 Review 
Documentation 
 
Osada, T. et al. Antihelminth compound niclosamide downregulates Wnt signaling and elicits 
antitumor responses in tumors with activating APC mutations. Cancer Res. (2011). 
doi:10.1158/0008-5472  
 
Ostrosky-Wegman, P. et al. Susceptibility to genotoxic effects of niclosamide in human 
peripheral lymphocytes exposed in vitro and in vivo. Mutat. Res. Lett. (1986). 
doi:10.1016/01657992(86)90015-1 
 
Park, D. W. et al. Role of JAK2–STAT3 in TLR2-Mediated Tissue Factor Expression. J Cell 
Biochem. (2013). doi: 10.1002/jcb.24472  
 
Perera, D. R. et al. Niclosamide treatment of cestodiasis. Clinical trials in the United States. Am J 
Trop Med Hyg. (1970). doi: 10.4269/ajtmh.1970.19.610  
 
Rehman, M. U. et al. Fabrication of Niclosamide loaded solid lipid nanoparticles: in vitro 
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Ren, X. et al. Identification of Niclosamide as a New Small-Molecule Inhibitor of the STAT3 
Signaling Pathway. ACS Med Chem Lett. (2010). doi: 10.1021/ml100146z  
 
Schuringa, J. J. et al. Constitutive Stat3, Tyr705, and Ser727 phosphorylation in acute myeloid 
leukemia cells caused by the autocrine secretion of interleukin-6. Blood. (2000). 
PMID:10845908   
 
Schweizer, M. T. et al. A phase I study of niclosamide in combination with enzalutamide in men 
with castration-resistant prostate cancer. PLoS One. (2018). doi: 10.1371/journal.pone.0198389  
 
Silva Borba, M. G. et al. Chloroquine diphosphate in two different dosages as adjunctive therapy 
of hospitalized patients with severe respiratory syndrome in the context of coronavirus 
(SARSCoV-2) infection: Preliminary safety results of a randomized, double-blinded, phase IIb 
clinical trial (CloroCovid-19 Study). medRxiv preprint. (2020). 
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Strufe, R. and Goennert, R. Experimentelle Untersuchungen mit N-(2'-Chlor-4'-nitrophenyl)-
5chlorsalicylamid, einem neuen Bandwurmmittel. / Experimental Studies on N-(2’-chloro-
4’nitrophenyl)-5-chlorosalicylamide, a New Taeniacide. Arzneimittel-Forsch. (1960) 10: 881-
884 (German / translated version)  
 
US Department of Health and Human Services. Common Terminology Criteria for Adverse 
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Wieland, A. et al. Anticancer Effects of Niclosamide in Human Glioblastoma. Clin Cancer Res. 
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Wilder-Smith, A. et al. Can we contain the COVID-19 outbreak with the same measures as for 
SARS? Lancet. Infect. Dis. (2020). doi:10.1016/S1473-3099(20)30129-8  
 
World Health Organization, Executive Summary, Safety Review of Niclosamide, Pyrantel, 
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Wu, C. J. et al. Inhibition of severe acute respiratory syndrome coronavirus replication by 
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Xu, J. et al. Broad Spectrum Antiviral Agent Niclosamide and Its Therapeutic Potential. ACS 
Infect. Dis. (2020). doi:10.1021/acsinfecdis.0c00052 
 
Yeh, H. H. et al. Upregulation of Tissue Factor by Activated Stat3 Contributes to Malignant 
Pleural Effusion Generation via Enhancing Tumor Metastasis and Vascular Permeability in Lung 
Adenocarcinoma. PLoS One. (2013). doi: 10.1371/journal.pone.0075287 
 
Yu , H. et al. Crosstalk between cancer and immune cells: role of STAT3 in the tumour 
microenvironment. Nat Rev Immunol. (2020). doi: 10.1038/nri1995 
 
Zhang, Y. Y. et al. Design and evaluation of injectable niclosamide nanocrystals prepared by wet 
media milling technique. Drug Dev Ind Pharm. (2015). doi: 10.3109/03639045.2014.954585 


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12. 
Appendices 


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Appendix 1: Abbreviations and Terms 
ANA  
ANA Therapeutics, Incorporated 
API  
Active pharmaceutical ingredient  
ARDS  
Acute respiratory distress syndrome  
AUC  
Area under curve  
BID  
Twice daily  
BSL 
Bio-safety Level 
CL  
Clearance  
Cmax  
Maximum serum concentration  
CoV  
Coronavirus  
COVID-19  
Coronavirus disease of 2019  
CYP  
Cytochrome P450  
DG  
Dose group  
DLT  
Dose-limiting toxicity  
DMC  
Data monitoring committee  
DMSO  
Dimethylsulfoxide  
DP  
Drug product  
DS  
Drug substance 
EC50  
Half-maximal effective concentration  
ECMO  
Extracorporeal membrane oxygenation  
IC50  
Half-maximal inhibitory concentration  
FDA  
Food and Drug Administration  
GI  
Gastrointestinal  
GLP  
Good laboratory practice  


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HDPE  
high-density polyethylene  
HTD  
Human therapeutic dose  
ICH  
International Council for Harmonization of Technical Requirements for 
Pharmaceuticals for Human Use 
IL  
Interleukin  
LD50  
Half-maximal lethal dose  
LLN 
Lower limit of normal 
MERS  
Middle East respiratory syndrome  
MRT  
Mean residence time  
NA 
Not applicable 
NDA  
New drug application  
NF-kB  
Nuclear factor kappa-light-chain-enhancer of activated B cells  
PBS  
Phosphate buffered saline  
PK  
Pharmacokinetic  
PO  
By mouth  
PVA  
Polyvinyl alcohol  
SAE  
Serious adverse event  
SARS  
Severe acute respiratory syndrome  
SBA  
Summary basis for approval  
STAT  
Signal transducers and activators of transcription  
SUSAR 
Suspected unexpected serious adverse reaction  
t1/2  
Elimination half-life  
TID  
Thrice daily  
tmax  
Time take to reach Cmax  


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TMEM16A  
Transmembrane member 16A  
TNF-α  
Tumor necrosis factor-alpha  
ULN 
Upper limit of normal 
VSS  
Volume of distribution at steady state  
WBC  
White blood cell  
WHO  
World Health Organization  


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Appendix 2: Clinical Laboratory Tests  
• The tests detailed in Table 4 will be performed by the local laboratory.  
• Additional tests may be performed at any time during the study as determined necessary 
by the Investigator or required by local regulations. 
Table 4 
 Protocol-Required Safety Laboratory Assessments 
Laboratory 
Assessments 
Parameters 
Hematology 
Platelet Count  
RBC Indices: 
MCV 
MCH 
% Reticulocytes 
WBC Count with 
Differential: 
Neutrophils 
Lymphocytes 
Monocytes 
Eosinophils 
Basophils 
RBC Count 
Hemoglobin 
Hematocrit 
Clinical 
Chemistry 
BUN 
Calcium 
Creatinine 
Glucose  
Potassium  
Sodium 
Total and direct 
bilirubin  
Total Protein 
Aspartate Aminotransferase (AST) 
Alanine Aminotransferase (ALT) 
Alkaline phosphatase 
Routine 
Urinalysis 
• Specific gravity 
• pH, glucose, protein, blood, ketones, bilirubin, urobilinogen, nitrite, 
leukocyte esterase by dipstick 
• Microscopic examination (if blood or protein is abnormal) 
Other 
Screening 
Tests 
• Urine alcohol and drug screens (to include at minimum: amphetamines, 
barbiturates, cocaine, opiates, cannabinoids and benzodiazepines) 
• Serum human chorionic gonadotropin (hCG) pregnancy test at Screening 
and urine pregnancy test on Day -1 for women of childbearing potential 
• Serology (HIV antibody, hepatitis B surface antigen [HBsAg], and 
hepatitis C virus antibody or specify other tests) 
Investigators must document their review of each laboratory safety report. 
 


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Appendix 3: Study Governance Considerations 
Regulatory and Ethical Considerations 
• This study will be conducted in accordance with the protocol and with the following: 
• Consensus ethical principles derived from international guidelines including the 
Declaration of Helsinki and Council for International Organizations of Medical 
Sciences (CIOMS) International Ethical Guidelines 
• Applicable ICH Good Clinical Practice (GCP) Guidelines 
• Applicable laws and regulations 
• The protocol, protocol amendments, ICF, Investigator Brochure, and other relevant 
documents (e.g., advertisements) must be submitted to an IRB/IEC by the Investigator 
and reviewed and approved by the IRB/IEC before the study is initiated.  
• Any amendments to the protocol will require IRB/IEC approval before implementation of 
changes made to the study design, except for changes necessary to eliminate an 
immediate hazard to study participants.  
• The Investigator will be responsible for the following: 
o Providing written summaries of the status of the study to the IRB/IEC annually or 
more frequently in accordance with the requirements, policies, and procedures 
established by the IRB/IEC 
o Notifying the IRB/IEC of SAEs or other significant safety findings as required by 
IRB/IEC procedures 
o Providing oversight of the conduct of the study at the site and adherence to 
requirements of 21 CFR, ICH guidelines, the IRB/IEC, European regulation 536/2014 
for clinical studies (if applicable), and all other applicable local regulations 
Financial Disclosure 
Investigators and sub-Investigators will provide the Sponsor with sufficient, accurate financial 
information as requested to allow the Sponsor to submit complete and accurate financial 
certification or disclosure statements to the appropriate regulatory authorities. Investigators are 
responsible for providing information on financial interests during the course of the study and for 
1 year after completion of the study. 
Informed Consent Process 
• The Investigator or his/her representative will explain the nature of the study to the 
participant or his/her legally authorized representative and answer all questions regarding 
the study.  
• Participants must be informed that their participation is voluntary. Participants will be 
required to sign a statement of informed consent that meets the requirements of 21 CFR 


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50, local regulations, ICH guidelines, Health Insurance Portability and Accountability 
Act (HIPAA) requirements, where applicable, and the IRB/IEC or study center.  
• The medical record must include a statement that written informed consent was obtained 
before the participant was enrolled in the study and the date the written consent was 
obtained. The authorized person obtaining the informed consent must also sign the ICF. 
• Participants must be re-consented to the most current version of the ICF(s) during their 
participation in the study.  
• A copy of the ICF(s) must be provided to the participant or the participant’s legally 
authorized representative.  
Data Protection 
• Participants will be assigned a unique identifier by the Sponsor. Any participant records 
or datasets that are transferred to the Sponsor will contain the identifier only; participant 
names or any information which would make the participant identifiable will not be 
transferred.  
• The participant must be informed that his/her personal study-related data will be used by 
the Sponsor in accordance with local data protection law. The level of disclosure must 
also be explained to the participant.  
• The participant must be informed that his/her medical records may be examined by 
Clinical Quality Assurance auditors or other authorized personnel appointed by the 
Sponsor, by appropriate IRB/IEC members, and by inspectors from regulatory 
authorities. 
Dissemination of Clinical Study Data 
A clinical study report will be prepared with reference to ICH Guidance E3 to include: 
• details of where the study was carried out, 
• dates of the start and completion of each period of the study, 
• details of the investigational product (IP) and a statement of production will be provided 
by ANA Therapeutics, Inc., 
• a statement confirming that the applicable IRB/IEC gave written approval for the study in 
accordance with local regulations, 
• a demographic listing for all participants, 
• a list of all AEs according to IP, 
• details of any occurrences which may be of significance to the study outcome, 
• details of all operations, calculations and transformations performed on the reported data, 
• the SAP and report will be produced by ANA Therapeutics, Inc. or their agents and will be 
incorporated into the final report, 
• all data from any withdrawn participant not included in the statistical analysis, 


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• a scientific interpretation of the results, 
• a description of the study methods used. 
Consideration will be given to any comments on a draft report. The report will incorporate the 
analytical and statistical results and methods produced by the Sponsor or their agents. A final 
report will be prepared to contain all those sections in the draft and a statement of compliance 
covering all the areas of the study conducted at the investigational site and the report, with GCP. 
The report will be issued under the Sponsor’s responsibility. 
Where required by the applicable regulatory requirements, an Investigator signatory will be 
identified for the approval of the clinical study report. The Investigator will be provided reasonable 
access to statistical tables, figures, and relevant reports and will have the opportunity to review 
complete study results. The Sponsor will also provide the Investigator with the full summary of 
study results. 
Data Quality Assurance 
All participant data relating to the study will be recorded on printed or electronic CRF unless 
transmitted to the Sponsor or designee electronically (e.g., laboratory data). The Investigator is 
responsible for verifying that data entries are accurate and correct by physically or electronically 
signing the CRF.  
The Investigator must maintain accurate documentation (source data) that supports the 
information entered in the CRF.  
The Investigator must permit study-related monitoring, audits, IRB/IEC review, and regulatory 
agency inspections and provide direct access to source data documents.  
The Sponsor or designee is responsible for the data management of this study including quality 
checking of the data.  
Study monitors will perform ongoing source data verification to confirm that data entered into 
the CRF by authorized site personnel are accurate, complete, and verifiable from source 
documents; that the safety and rights of participants are being protected; and that the study is 
being conducted in accordance with the currently approved protocol and any other study 
agreements, ICH GCP, and all applicable regulatory requirements.  
Records and documents, including signed ICFs, pertaining to the conduct of this study must be 
retained by the Investigator for 5 years after study completion unless local regulations or 
institutional policies require a longer retention period. No records may be destroyed during the 
retention period without the written approval of the Sponsor. No records may be transferred to 
another location or party without written notification to the Sponsor.  


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Source Documents 
Source documents provide evidence for the existence of the participant and substantiate the 
integrity of the data collected. Source documents are filed at the Investigator’s site. 
Data reported on the CRF or entered in the eCRF that are transcribed from source documents 
must be consistent with the source documents or the discrepancies must be explained. The 
Investigator may need to request previous medical records or transfer records, depending on the 
study. Also, current medical records must be available. 
Study and Site Closure 
The Sponsor designee reserves the right to close the study site or terminate the study at any time 
for any reason at the sole discretion of the Sponsor. Study sites will be closed upon study 
completion. A study site is considered closed when all required documents and study supplies 
have been collected and a study-site closure visit has been performed. 
The Investigator may initiate study-site closure at any time, provided there is reasonable cause 
and sufficient notice is given in advance of the intended termination. 
Reasons for the early closure of a study site by the Sponsor or Investigator may include but are 
not limited to: 
• Failure of the Investigator to comply with the protocol, the requirements of the IRB/IEC 
or local health authorities, the Sponsor’s procedures, or GCP guidelines 
• Inadequate recruitment of participants by the Investigator 
• Discontinuation of further study treatment development 
 
 


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Appendix 4: Adverse Events: Definitions and Procedures for Recording, 
Evaluating, Follow-up, and Reporting 
Definition of an Adverse Event (AE) 
AE Definition 
• 
An AE is any untoward medical occurrence in a patient or clinical study participant, 
temporally associated with the use of study treatment, whether or not considered 
related to the study treatment. 
• 
NOTE: An AE can therefore be any unfavorable and unintended sign (including an 
abnormal laboratory finding), symptom, or disease (new or exacerbated) temporally 
associated with the use of study treatment. 
 
Events Meeting the AE Definition  
• 
Any abnormal laboratory test results (hematology, clinical chemistry, or urinalysis) 
or other safety assessments (e.g., ECG, radiological scans, vital signs 
measurements), including those that worsen from baseline, considered clinically 
significant by the Investigator per Appendix 6 (i.e., not related to progression of 
underlying disease). 
• 
Exacerbation of a chronic or intermittent pre-existing condition including either an 
increase in frequency and/or intensity of the condition. 
• 
New conditions detected or diagnosed after study treatment administration even 
though it may have been present before the start of the study. 
• 
Signs, symptoms, or the clinical sequelae of a suspected drug-drug interaction. 
• 
Signs, symptoms, or the clinical sequelae of a suspected overdose of either study 
treatment or a concomitant medication. Overdose per se will not be reported as an 
AE/SAE unless it is an intentional overdose taken with possible suicidal/self-
harming intent. Such overdoses should be reported regardless of sequelae. 
 
Events NOT Meeting the AE Definition  
• 
Any clinically significant abnormal laboratory findings or other abnormal safety 
assessments which are associated with the underlying disease, unless judged by the 
Investigator to be more severe than expected for the participant’s condition. 


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• 
The disease/disorder being studied or expected progression, signs, or symptoms of 
the disease/disorder being studied, unless more severe than expected for the 
participant’s condition. 
• 
Medical or surgical procedure (e.g., endoscopy, appendectomy): the condition that 
leads to the procedure is the AE. 
• 
Situations in which an untoward medical occurrence did not occur (social and/or 
convenience admission to a hospital). 
• 
Anticipated day-to-day fluctuations of pre-existing disease(s) or condition(s) present 
or detected at the start of the study that do not worsen. 
 
Definition of SAE 
If an event is not an AE per definition above, then it cannot be an SAE even if serious conditions 
are met (e.g., hospitalization for signs/symptoms of the disease under study, death due to 
progression of disease). 
A SAE is defined as any untoward medical occurrence that, at any dose: 
a. Results in death 
b. Is life-threatening 
The term 'life-threatening' in the definition of 'serious' refers to an event in which the 
participant was at risk of death at the time of the event. It does not refer to an event, 
which hypothetically might have caused death, if it were more severe. 
c. 
Requires inpatient hospitalization or prolongation of existing hospitalization 
In general, hospitalization signifies that the participant has been detained (usually 
involving at least an overnight stay) at the hospital or emergency ward for observation 
and/or treatment that would not have been appropriate in the physician’s office or 
outpatient setting. Complications that occur during hospitalization are AEs. If a 
complication prolongs hospitalization or fulfills any other serious criteria, the event is 
serious. When in doubt as to whether “hospitalization” occurred or was necessary, the AE 
should be considered serious. 
Hospitalization for elective treatment of a pre-existing condition that did not worsen from 
baseline is not considered an AE. 
d. Results in persistent disability/incapacity 


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• 
The term disability means a substantial disruption of a person’s ability to conduct 
normal life functions. 
• 
This definition is not intended to include experiences of relatively minor medical 
significance such as uncomplicated headache, nausea, vomiting, diarrhea, influenza, 
and accidental trauma (e.g., sprained ankle) which may interfere with or prevent 
everyday life functions but do not constitute a substantial disruption. 
e. 
Is a congenital anomaly/birth defect 
f. 
Other situations: 
• 
Medical or scientific judgment should be exercised in deciding whether SAE 
reporting is appropriate in other situations such as important medical events that may 
not be immediately life-threatening or result in death or hospitalization but may 
jeopardize the participant or may require medical or surgical intervention to prevent 
one of the other outcomes listed in the above definition. These events should usually 
be considered serious. 
Examples of such events include invasive or malignant cancers, intensive treatment 
in an emergency room or at home for allergic bronchospasm, blood dyscrasias or 
convulsions that do not result in hospitalization, or development of drug dependency 
or drug abuse. 
Recording and AE and/or SAE 
AE and SAE Recording 
• 
When an AE/SAE occurs, it is the responsibility of the Investigator to review all 
documentation (e.g., hospital progress notes, laboratory reports, and diagnostics 
reports) related to the event. 
• 
The Investigator will then record all relevant AE/SAE information in the CRF. 
• 
It is not acceptable for the Investigator to send photocopies of the participant’s 
medical records to ANA Therapeutics in lieu of completion of the AE/SAE CRF 
page. 
• 
There may be instances when copies of medical records for certain cases are 
requested by ANA Therapeutics. In this case, all participant identifiers, with the 
exception of the participant number, will be redacted on the copies of the medical 
records before submission to ANA Therapeutics. 


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• 
The Investigator will attempt to establish a diagnosis of the event based on signs, 
symptoms, and/or other clinical information. Whenever possible, the diagnosis (not 
the individual signs/symptoms) will be documented as the AE/SAE. 
Assessment of Intensity 
The Investigator will make an assessment of intensity for each AE and SAE reported 
during the study and assign it to 1 of the following categories:  
• 
Mild: An event that is easily tolerated by the participant, causing minimal discomfort 
and not interfering with everyday activities. 
• 
Moderate: An event that causes sufficient discomfort and interferes with normal 
everyday activities. 
• 
Severe: An event that prevents normal everyday activities. An AE that is assessed as 
severe should not be confused with a SAE. Severe is a category utilized for rating the 
intensity of an event; and both AEs and SAEs can be assessed as severe. 
An event is defined as ‘serious’ when it meets at least 1 of the predefined outcomes 
as described in the definition of an SAE, NOT when it is rated as severe. 
 
Assessment of Causality 
• 
The Investigator is obligated to assess the relationship between study treatment and 
each occurrence of each AE/SAE. 
• 
A "reasonable possibility" of a relationship conveys that there are facts, evidence, 
and/or arguments to suggest a causal relationship, rather than a relationship cannot 
be ruled out. 
• 
The Investigator will use clinical judgment to determine the relationship. 
• 
Alternative causes, such as underlying disease(s), concomitant therapy, and other 
risk factors, as well as the temporal relationship of the event to study treatment 
administration will be considered and investigated. 
• 
The Investigator will also consult the Investigator’s Brochure (IB) and/or Product 
Information, for marketed products, in his/her assessment. 
• 
For each AE/SAE, the Investigator must document in the medical notes that he/she 
has reviewed the AE/SAE and has provided an assessment of causality. 


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• 
There may be situations in which an SAE has occurred and the Investigator has 
minimal information to include in the initial report to ANA Therapeutics. However, 
it is very important that the Investigator always make an assessment of 
causality for every event before the initial transmission of the SAE data to ANA 
Therapeutics. 
• 
The Investigator may change his/her opinion of causality in light of follow-up 
information and send a SAE follow-up report with the updated causality assessment. 
• 
The causality assessment is one of the criteria used when determining regulatory 
reporting requirements. 
 
Follow-up of AEs and SAEs 
• 
The Investigator is obligated to perform or arrange for the conduct of supplemental 
measurements and/or evaluations as medically indicated or as requested by ANA 
Therapeutics to elucidate the nature and/or causality of the AE or SAE as fully as 
possible. This may include additional laboratory tests or investigations, 
histopathological examinations, or consultation with other health care professionals. 
• 
If a participant dies during participation in the study or during a recognized follow-
up period, the Investigator will provide ANA Therapeutics with a copy of any post-
mortem findings including histopathology. 
• 
New or updated information will be recorded in the originally completed CRF. 
• 
The Investigator will submit any updated SAE data to the ANA Therapeutics within 
24 hours of receipt of the information. 
 
Reporting of SAEs 
SAE Reporting to ANA Therapeutics via an Electronic Data Collection Tool 
• 
The primary mechanism for reporting an SAE to ANA Therapeutics will be the 
electronic data collection tool. 
• 
If the electronic system is unavailable for more than 24 hours, then the site will use 
the paper SAE data collection tool (see next section). 
• 
The site will enter the SAE data into the electronic system as soon as it becomes 
available. 


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• 
After the study is completed at a given site, the electronic data collection tool will be 
taken off-line to prevent the entry of new data or changes to existing data. 
• 
If a site receives a report of a new SAE from a study participant or receives updated 
data on a previously reported SAE after the electronic data collection tool has been 
taken off-line, then the site can report this information on a paper SAE form (see 
next section) or by telephone. 
• 
Contacts for SAE reporting can be found in the Study Manual. 
 
SAE Reporting via Paper CRF 
• 
Facsimile transmission of the SAE paper CRF is the preferred method to transmit 
this information to the SAE coordinator. 
• 
In rare circumstances and in the absence of facsimile equipment, notification by 
telephone is acceptable with a copy of the SAE data collection tool sent by overnight 
mail or courier service. 
• 
Initial notification via telephone does not replace the need for the Investigator to 
complete and sign the SAE CRF pages within the designated reporting time frames. 
• 
Contacts for SAE reporting can be found in the Study Manual. 
 
 


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Appendix 5: Contraceptive Guidance and Collection of Pregnancy 
Information 
Definitions 
Woman of Childbearing Potential (WOCBP) 
A woman is considered fertile following menarche and until becoming post-menopausal unless 
permanently sterile (see below). 
Women in the following categories are not considered WOCBP 
1. Premenarchal 
2. Premenopausal female with 1 of the following: 
• Documented hysterectomy 
• Documented bilateral salpingectomy 
• Documented bilateral oophorectomy 
Note:  Documentation can come from the site personnel’s: review of the participant’s 
medical records, medical examination, or medical history interview. 
 
3. Postmenopausal female 
• A postmenopausal state is defined as no menses for 12 months without an alternative 
medical cause. A high follicle stimulating hormone (FSH) level in the 
postmenopausal range may be used to confirm a postmenopausal state in women not 
using hormonal contraception or hormonal replacement therapy (HRT). However, in 
the absence of 12 months of amenorrhea, a single FSH measurement is insufficient.  
• Females on HRT and whose menopausal status is in doubt will be required to use one 
of the non-hormonal highly effective contraception methods if they wish to continue 
their HRT during the study. Otherwise, they must discontinue HRT to allow 
confirmation of postmenopausal status before study enrollment. 
Contraception Guidance 
Male participants 
• Male participants with female partners of childbearing potential are eligible to 
participate if they agree to ONE of the following [during the protocol-defined time 
frame in Section 6.1]:  


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Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
Are abstinent from penile-vaginal intercourse as their usual and preferred lifestyle (abstinent on a 
long term and persistent basis) and agree to remain abstinent 
• Agree to use a male condom plus partner use of a contraceptive method with a failure 
rate of <1% per year as described in Table 5 when having penile-vaginal intercourse 
with a woman of childbearing potential who is not currently pregnant 
Men with a pregnant or breastfeeding partner must agree to remain abstinent from penile-vaginal 
intercourse or use a male condom during each episode of penile penetration [during the protocol-
defined time frame]  
Refrain from donating sperm for the duration of the study and for 30 days after the last dose of 
study treatment. 
Female participants 
Female participants of childbearing potential are eligible to participate if they agree to use a 
highly effective method of contraception consistently and correctly as described in Table 5. 
 
Table 5: 
 Highly Effective Contraceptive Methods 
Highly Effective Contraceptive Methods That Are User Dependent a  
Failure rate of <1% per year when used consistently and correctly.  
Combined (estrogen and progestogen containing) hormonal contraception associated with 
inhibition of ovulationb  
• Oral  
• Intravaginal  
• Transdermal  
Progestogen only hormonal contraception associated with inhibition of ovulation 
• Oral  
• Injectable  
Highly Effective Methods That Are User Independent a  
Implantable progestogen only hormonal contraception associated with inhibition of ovulationb 
• Intrauterine device (IUD) 
• Intrauterine hormone-releasing system (IUS) 
Bilateral tubal occlusion 
Vasectomized partner  
A vasectomized partner is a highly effective contraception method provided that the partner is 
the sole male sexual partner of the WOCBP and the absence of sperm has been confirmed. If 
not, an additional highly effective method of contraception should be used.  
Sexual abstinence  


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Status: Final v2.0; Date 09Jan2021 
 
Sexual abstinence is considered a highly effective method only if defined as refraining from 
heterosexual intercourse during the entire period of risk associated with the study treatment. 
The reliability of sexual abstinence needs to be evaluated in relation to the duration of the 
study and the preferred and usual lifestyle of the participant.  
NOTES:  
a) Typical use failure rates may differ from those when used consistently and correctly. Use 
should be consistent with local regulations regarding the use of contraceptive methods for 
participants participating in clinical studies.  
b) Hormonal contraception may be susceptible to interaction with the study treatment, which 
may reduce the efficacy of the contraceptive method. In this case, two highly effective 
methods of contraception should be utilized during the treatment period and for at least 30 
days after the last dose of study treatment  
 
Pregnancy Testing 
• 
WOCBP should only be included after a confirmed menstrual period and a negative highly 
sensitive pregnancy test. 
• 
Pregnancy testing will be performed whenever a menstrual cycle is missed or when 
pregnancy is otherwise suspected  
Collection of Pregnancy Information  
Male participants with partners who become pregnant  
• 
The Investigator will attempt to collect pregnancy information on any male participant’s 
female partner who becomes pregnant while the male participant is in this study. This applies 
only to male participants who receive active study treatment. 
• 
After obtaining the necessary signed informed consent from the pregnant female partner 
directly, the Investigator will record pregnancy information on the appropriate form and 
submit it to the Sponsor within 24 hours of learning of the partner’s pregnancy. The female 
partner will also be followed to determine the outcome of the pregnancy. Information on the 
status of the mother and child will be forwarded to the Sponsor. Generally, the follow-up will 
be no longer than 6 to 8 weeks following the estimated delivery date. Any termination of the 
pregnancy will be reported regardless of fetal status (presence or absence of anomalies) or 
indication for the procedure.  
Female Participants who become pregnant  


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Protocol ANA001 
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Status: Final v2.0; Date 09Jan2021 
 
• 
The Investigator will collect pregnancy information on any female participant who becomes 
pregnant while participating in this study. Information will be recorded on the appropriate 
form and submitted to the Sponsor within 24 hours of learning of a participant's pregnancy. 
The participant will be followed to determine the outcome of the pregnancy. The Investigator 
will collect follow-up information on the participant and the neonate and the information will 
be forwarded to the Sponsor. Generally, follow-up will not be required for longer than 6 to 8 
weeks beyond the estimated delivery date. Any termination of pregnancy will be reported, 
regardless of fetal status (presence or absence of anomalies) or indication for the procedure. 
• 
While pregnancy itself is not considered to be an AE or SAE, any pregnancy complication or 
elective termination of a pregnancy will be reported as an AE or SAE. A spontaneous abortion 
is always considered to be an SAE and will be reported as such. Any post-study pregnancy-
related SAE considered reasonably related to the study treatment by the Investigator will be 
reported to the Investigator as described in Section 9.2. While the Investigator is not obligated 
to actively seek this information in former study participants, he or she may learn of an SAE 
through spontaneous reporting.  
• 
Any female participant who becomes pregnant while participating in the study will be 
withdrawn from the study 
 
 


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Protocol ANA001 
Page 68 of 69 
 
Status: Final v2.0; Date 09Jan2021 
 
Appendix 6: Potentially Clinically Significant Laboratory Criteria 
(Local Laboratory Results) 
Parameter 
SI Unit 
Lower 
Limit 
Higher Limit 
% Decrease 
from Bio-
safety Level 
(BSL) 
% Increase 
from BSL 
CHEMISTRY 
Blood Urea Nitrogen 
(BUN) 
mmol/L 
NA 
>1.5 *ULN 
NA 
>100 
Creatinine 
mg/dL 
NA 
>1.5 *ULN 
NA 
>50 
Sodium 
mmol/L 
<0.85 *LLN 
>1.1 *ULN 
>10 
>10 
Potassium 
mol/L 
<0.8 *LLN 
>1.2 *ULN 
>20 
>20 
Chloride 
mmol/L 
<0.8 *LLN 
>1.2 *ULN 
>20 
>20 
Carbon Dioxide 
(Bicarbonate) 
mmol/L 
<0.7 *LLN 
>1.3 *ULN 
>40 
>40 
Glucose 
mmol/L 
<0.6 *LLN 
>3.0 *ULN 
>40 
>200 
Total Bilirubin 
mg/dL 
NA 
>2.5 *ULN 
NA 
>150 
Direct Bilirubin 
mg/dL 
NA 
>1.5 *ULN 
NA 
>150 
Alkaline Phosphatase 
U/L 
<0.5 *LLN 
>2.0 *ULN 
>80 
>100 
Aspartate 
Aminotransferase 
(AST) 
U/L 
NA 
>3.0 *ULN 
NA 
>200 
Alanine 
Aminotransferase 
(ALT) 
U/L 
NA 
>3.0 *ULN 
NA 
>200 
HEMATOLOGY 
Hematocrit 
Ratio 
<0.8 *LLN 
>1.3 *ULN 
>20 
>30 
Hemoglobin 
g/L 
<0.8 *LLN 
>1.3 *ULN 
>20 
>30 
Platelet Count 
109/L 
<0.65 *LLN 
>1.5 *ULN 
>50 
>100 
White Blood Cell Count 
109/L 
<0.65 *LLN 
>1.6 *ULN 
>60 
>100 
Neutrophils, absolute 
cell count 
109/L 
<0.65 *LLN 
>1.6 *ULN 
>75 
>100 


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Status: Final v2.0; Date 09Jan2021 
 
Parameter 
SI Unit 
Lower 
Limit 
Higher Limit 
% Decrease 
from Bio-
safety Level 
(BSL) 
% Increase 
from BSL 
Lymphocytes, absolute 
count 
109/L 
<0.25 *LLN 
>1.5 *ULN 
>75 
>100 
Eosinophils, absolute 
count 
109/L 
NA 
>4.0 *ULN 
NA 
>300 
Monocytes, absolute 
count 
109/L 
NA 
>4.0 *ULN 
NA 
>300 
Basophils, absolute 
count 
109/L 
NA 
>4.0 *ULN 
NA 
>300 
COAGULATION 
Partial Thromboplastin 
Time (aPTT) 
sec 
<0.5 *LLN 
>2.0 *ULN 
>100 
>100 
Prothrombin Time (PT) 
sec 
<0.5 *LLN 
>2.0 *ULN 
>100 
>100 
International 
Normalized Ratio (INR) 
Ratio 
<0.5 *LLN 
>2.0 *ULN 
>100 
>100 
LLN:  Lower limit of normal value provided by the local laboratory 
ULN: Upper limit of normal value provided by the local laboratory 
 

SAP_001.pdf: 
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Page 1 of 27 
  
 
 
 
STATISTICAL ANALYSIS PLAN 
 
 
 
AUTHOR(S): 
Sulagna Sarkar  
PROJECT MANAGER: 
Neil Patel  
 
 
SAP VERSION: 
Draft 1.0 
 
DATE OF SAP VERSION 
4 December 2020  
 
 
 
 
SPONSOR: 
ANA Therapeutics, Inc. 
602 Bainbridge St.  
Foster City, CA 94404 
 
STUDY TITLE: 
A Randomized, Double-Blind, Single and Multiple 
Ascending Dose Study to Assess the Safety and 
Pharmacokinetics of ANA001 in Healthy Adults 
 
PHASE OF STUDY: 
Phase 1 
PROTOCOL NUMBER: 
ANA001-002 
PROTOCOL VERSION/DATE: Version 1.0; 22 October 2020  
 
 
 
 
 
 
 
 
 
 


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APPROVALS 
 
Name and Title 
Signature 
Date 
 
Doug Rank, MD 
VP of Clinical Development 
ANA Therapeutics, Inc. 
 
 
 
 
Kalyan Ghosh, Ph.D. 
Biostatistician 
Inference, Inc 
 
 
 


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TABLE OF CONTENTS 
1. 
ABBREVIATIONS ......................................................................................................5 
2. 
INTRODUCTION ........................................................................................................6 
2.1. 
Responsibilities .............................................................................................................6 
2.2. 
Timing of Analyses .......................................................................................................6 
2.3. 
Interim Analysis ............................................................................................................6 
3. 
STUDY OBJECTIVES AND ENDPOINTS ................................................................7 
3.1. 
Study Objectives ...........................................................................................................7 
3.2. 
Endpoints ......................................................................................................................7 
3.2.1. 
Primary Outcome Measures .........................................................................................7 
3.2.2. 
Secondary Endpoints ....................................................................................................7 
4. 
STUDY DESIGN .........................................................................................................9 
4.1. 
Sample Size Justification ............................................................................................10 
4.2. 
Schedule of Activities (SoA) ......................................................................................10 
5. 
ANALYSIS SETS ......................................................................................................14 
6. 
GENERAL ASPECTS OF THE STATISTICAL ANALYSIS..................................15 
6.1. 
Analysis of the Two Parts ...........................................................................................15 
6.2. 
Key Definitions ...........................................................................................................15 
6.3. 
Visit Windows and Time Points .................................................................................15 
6.4. 
Multiplicity Issues ......................................................................................................15 
6.5. 
Subgroup Analyses .....................................................................................................15 
6.6. 
Missing Data ...............................................................................................................15 
7. 
DEMOGRAPHIC AND BASELINE CHARACTERISTICS ....................................17 
7.1. 
Subject Disposition and Populations ..........................................................................17 
7.2. 
Demographic and Baseline Characteristics ................................................................17 
7.3. 
Medical History ..........................................................................................................17 
7.4. 
Prior and Concomitant Medication .............................................................................17 
8. 
PHARMACOKINETIC ANALYSIS .........................................................................19 
8.1. 
Handling Missing or Non-Quantifiable Data .............................................................19 
8.2. 
Pharmacokinetic Concentration Analysis ...................................................................19 
8.3. 
Pharmacokinetic Parameter Analysis .........................................................................19 
9. 
SAFETY ANALYSIS ................................................................................................22 


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9.1. 
Adverse Events ...........................................................................................................22 
9.2. 
Clinical Laboratory Assessments ...............................................................................22 
9.3. 
Vital Signs ..................................................................................................................23 
9.4. 
ECG ............................................................................................................................23 
9.5. 
Physical Examination .................................................................................................23 
10. 
ANALYSIS CONVENTIONS ...................................................................................24 
11. 
REFERENCES ...........................................................................................................26 
12. 
TABLES, LISTINGS, AND FIGURES .....................................................................27 
 


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ABBREVIATIONS 
 
Abbreviation or Special 
Term 
Explanation  
AE 
Adverse event  
ANA 
ANA Therapeutics, Incorporated  
AUC  
Area under curve  
BID  
Twice daily   
BLQ 
Below limit of quantification  
CL  
Clearance  
Cmax  
Maximum serum concentration  
COVID-19  
Coronavirus disease of 2019  
CV 
Coefficient of variation  
DMC  
Data monitoring committee  
FDA  
Food and Drug Administration  
ICF 
Informed consent form  
ICH  
International Council for Harmonization of Technical 
Requirements for Pharmaceuticals for Human Use  
MedDRA 
Medical Dictionary for Regulatory Activities  
NCA 
Non-compartmental analysis  
NDA  
New drug application  
PK  
Pharmacokinetic  
PT 
Preferred term  
PVA  
Polyvinyl alcohol  
SAE  
Serious adverse event  
SD 
Standard deviation  
SoA 
Schedule of Activities  
SOC 
System organ class 
t1/2  
Elimination half-life  
TEAE 
Treatment-emergent adverse event  
TID  
Thrice daily  
tmax   
Time take to reach Cmax  
TNF-α  
Tumor necrosis factor-alpha  
VSS  
Volume of distribution at steady state  
WBC  
White blood cell  
WHO  
World Health Organization  
 
 


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1. 
INTRODUCTION 
The purpose of this plan is to prospectively outline in detail the data derivations, statistical methods 
and presentations of data so that valid conclusions can be reached to address the study objectives 
outlined in the protocol ANA001-002, dated 22 October 2020. 
 
The planned analyses identified in this statistical analysis plan (SAP) may be included in 
regulatory submissions and/or future manuscripts.  Exploratory analyses, not identified in this 
SAP, may be performed to support the clinical development program.  Any post-hoc or unplanned 
analyses that are performed but not identified in this SAP will be clearly identified in the clinical 
study report (CSR). 
 
1.1. 
Responsibilities 
Inference, Inc will perform the statistical analyses for all clinical data collected. Inference, Inc is 
responsible for production and quality control of all tables, figures and listings. 
 
1.2. 
Timing of Analyses 
Data from the SAD Cohorts S1 through S3 will be summarized and evaluated to determine the 
doses and schedule to be used in a potential fourth SAD cohort and the MAD BID or TID cohorts. 
Dosing for the potential fourth SAD Cohort S4 will be based on both safety and PK data.  
The final analysis will be performed after all subjects complete the study and the database is 
locked. 
 
1.3. 
Interim Analysis 
The interim analysis based on data from the SAD Cohorts S1 through S3 will be done using 
aggregate summaries to preserve blinding. The analysis will include listing of safety data, 
including adverse events. Serious and drug-related adverse events will be taken into in determining 
the dosing in the MAD part of the study. It may also involve evaluation of laboratory safety data. 
These analyses will be based on the Safety Analysis. 
For evaluating the pharmacokinetics of single dose ANA001, plots of mean (SD) plasma 
concentration by time will be presented for ANA001 by cohort/dose in linear and semi-logarithmic 
scales.  Pharmacokinetic parameters will also be calculated by each cohort/dose using the methods 
describes in Section 8, but will use nominal times instead of actual sampling times. 
 


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2. 
STUDY OBJECTIVES AND ENDPOINTS 
2.1. 
Study Objectives 
 
Primary Objectives: 
 
• 
To assess safety and tolerability of single oral daily doses of ANA001 (1,000 mg, 
2,000 mg and 3,000 mg)  
• 
To assess safety and tolerability of multiple daily oral doses (twice daily [BID] or 
thrice daily [TID]) of ANA001 for 7 days  
 
Secondary Objectives: 
 
• 
To assess the PK of single and multiple doses of ANA001  
2.2. 
Endpoints 
2.2.1. 
Primary Outcome Measures 
The primary outcome measures are the following safety and tolerability endpoints:  
• 
Incidence of: 
 
Treatment-emergent adverse events (AEs) 
 
Treatment-emergent serious AEs (SAEs) 
 
Study drug discontinuation due to a TEAE (MAD portion only) 
• 
Use of concomitant medications 
• 
Incident of treatment-emergent clinical laboratory tests abnormality 
• 
Change from baseline by visit in: 
 
Clinical laboratory tests 
 
Vital signs 
 
Electrocardiograms (ECG) 
 
Physical examinations  
2.2.2. 
Secondary Endpoints 
The secondary outcome measures are the following pharmacokinetic endpoints:  
• 
AUC(0-t) 
• 
AUC(0-last) 
• 
AUC(0-∞) (SAD only) 
• 
Cmax 
• 
tmax 


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• 
t1/2 
• 
CL/F 
• 
Vz/F  
 


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3. 
STUDY DESIGN 
This is a randomized, double blind study to be conducted in two parts: single ascending dose 
(SAD) and multiple ascending dose (MAD).  
 
SAD  
 
Eligible participants will be randomly assigned to active drug (ANA001) or placebo in an 8:2 ratio. 
On Day 1, following an overnight fast of at least 10 hours, participants will receive their assigned 
treatment (1000 mg, 2000 mg, or 3000 mg of ANA001) with a standardized light meal.  
 
A baseline ECG will be performed at Screening and at 3 hours post-dose. Blood pressure and heart 
rate will be measured every 8 hours while confined to the clinic. Blood for PK will be collected 
pre-dose and at 0.5, 1, 2, 4, 6, 8, 10, 12, and 24 hours post-dose. Participants will be discharged on 
Day 2 following the 24-hour PK sample and completion of clinical laboratory tests and physical 
examination. They will return on Day 7 (±2) for follow-up procedures including physical 
examination, laboratory tests, and assessment of AEs and concomitant medication use.  
 
SAD study design: 
Cohort S1: 1000 mg (n=8) or placebo (n=2), single dose  
Cohort S2: 2000 mg (n=8) or placebo (n=2), single dose  
Cohort S3: 3000 mg (n=8) or placebo (n=2), single dose  
Cohort S4: xx mg (n=8) or placebo (n=2) (optional; to be determined)  
 
MAD  
 
Eligible participants will be randomly assigned to active drug (ANA001) or placebo in a 10:2 ratio 
on a BID (q12h) or TID (q8h) schedule, to be determined based on the PK results of the SAD 
cohorts. Total daily doses are anticipated to range between 2000 mg and 4500 mg of ANA001. On 
Days 1 through 7, participants will receive their assigned treatment in the morning with a 
standardized light meal following an overnight fast of at least 10 hours. Afternoon or evening 
doses will be given with a light snack. A baseline ECG will be performed at Screening and at 3 
hours after the second dose on Day 7. Blood pressure and heart rate will be measured every 8 hours 
while confined to the clinic.  
 
During BID dosing, blood for PK will be collected on Days 1 and 7 at pre-dose and at 0.5, 1, 2, 4, 
6, 8, 10, and 12 hours post-dose. Blood samples will also be collected on Days 2, 4, 6, and 8 at 
pre-dose and before the 12 hour dose on Days 2, 4, and 6.  
 
During TID dosing, blood for PK will be collected on Days 1 and 7 at pre-dose and at 0.5, 1, 2, 4, 
6, and 8 hours post-dose. Blood samples will also be collected on Days 2, 4, 6, and 8 at pre-dose, 
before the 8 hour dose on Days 2, 4, and 6, and before the 16 hour dose on Day 6.  
 
Participants will be discharged on Day 8 following the last PK sample and completion of clinical 
laboratory tests and physical examination. They will return on Day 15 (±2) for follow-up 


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procedures including physical examination, laboratory tests, and assessment of AEs and 
concomitant medication use. 
 
MAD study design: 
Cohort M1: xx mg (n=9) or placebo (n=3) BID or TID for 7 days  
Cohort M2: xx mg (n=9) or placebo (n=3) BID or TID for 7 days  
Cohort M3: xx mg (n=9) or placebo (n=3) BID or TID for 7 days 
 
3.1. 
Sample Size Justification 
 
Sample size was not based on statistical considerations. A minimum of 30 participants will be 
enrolled in the SAD part and up to 36 participants in the MAD part. At least 3 participants of each 
sex will be included in each cohort. 
 
3.2. 
Schedule of Activities (SoA) 
 
 The following assessments will be performed in this study:   
 
 


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4. ANALYSIS SETS  
Screened participants will be defined as any participant who has signed the ICF. Screened 
participants who meet all eligibility criteria will be considered enrolled into the study and eligible 
to receive the single dose of ANA001. 
 
There will be two analysis populations in this study:  
 
Analysis Population 
Description 
Enrolled 
All participants who sign the ICF 
 
Safety Population 
All participants randomly assigned to study treatment and who take 
at any amount of study treatment. Participants will be analyzed 
according to the treatment they actually received. 
 
Pharmacokinetic (PK) 
Population 
All participants randomly assigned to study treatment, who take at 
least 1 dose of study treatment, and contribute at least 1 post-dose 
evaluable PK sample. Participants will be analyzed according to the 
dose they actually received. 
 
 


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5. GENERAL ASPECTS OF THE STATISTICAL ANALYSIS 
5.1. 
Analysis of the Two Parts 
The analyses for the SAD and MAD parts will be presented separately. 
 
5.2. 
Key Definitions 
The Study Day is the day relative to the day of study drug administration (Day 1). 
Unless otherwise specified, Baseline is the last non-missing observation before the administration 
of study drug. The treatment period in both the SAD and MAD parts of the study is defined as 
Days 1-7 for subjects dosed with study drug.  
 
5.3. 
Visit Windows and Time Points 
There are no plans to derive visit windows, and visits will be used in the analyses as reported on 
the eCRF.  
5.4. 
Multiplicity Issues 
For this early phase study, no multiplicity adjustment will be necessary as no comparisons are 
being made.  
 
5.5. 
Subgroup Analyses 
Due to the small size of this study, no subgroup analyses are planned.   
 
5.6. 
Missing Data 
For prior and concomitant medication summaries, if the medication start date is completely 
missing then the medication will be considered both prior and concomitant unless it can be 
determined that the medication end date occurred prior to the study drug administration.  If the 
medication start date is partially missing and the partial date is not sufficient to determine if the 
medication was taken after study drug administration then the medication will be considered both 
prior and concomitant for the study unless the partial date is clearly after the date of study drug 
administration (in which case it will be considered concomitant only) or the medication end date 
is prior to study drug administration (in which case it is prior only). 
Completely missing or partially missing adverse event onset dates will be imputed as follows in 
case due diligence to obtain accurate adverse event information fails: 
• If the adverse event start date is completely missing then the adverse event will be 
considered treatment emergent unless it can be determined that the adverse event end date 


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occurred prior to administration of study drug.  If this is the case, the adverse event will 
not be considered treatment emergent. 
• If the adverse event start date is partially missing and the partial date is not sufficient to 
determine if the event occurred after the administration of study drug, then the adverse 
event will be considered treatment emergent unless it can be determined that the adverse 
event end date occurred prior to the start of the study.  
Imputation of missing PK data are detailed in the PK section (section 8.1). 
No other missing data will be imputed. 


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6. 
DEMOGRAPHIC AND BASELINE CHARACTERISTICS  
6.1. 
Subject Disposition and Populations 
 
Summary tables for subject disposition will be presented for all enrolled subjects, by cohort/dose 
and overall. The subject disposition summary will contain the numbers of subjects who signed 
informed consent, failed screening, were randomized, completed the study, and discontinued 
during the study.   
The primary reasons for premature discontinuation during the treatment period will be summarized 
for the Safety Population. 
Number of subjects included in each analysis population will also be displayed. 
Listings will be presented on subject disposition and protocol violations. 
 
6.2. 
Demographic and Baseline Characteristics   
 
All subjects in the Safety Population will be used to summarize the demographic and baseline 
characteristics with respect to key variables such as age, sex, race, height, weight, and BMI by 
cohort/dose and overall. Summaries for other variables may be provided if needed.  
 
Continuous variables will be summarized with descriptive statistics: n, mean, standard deviation, 
median, minimum and maximum value.  Categorical variables will be summarized with the 
number and percentages for each cohort.  
 
Demographic and baseline characteristics such as COVID-19 screening, pregnancy test for 
WOCBP, urine alcohol and drug, and serology (HIV, HBV, HCV) tests will be listed for each 
subject.   
 
6.3. 
Medical History 
 
Medical history including existing diseases will be coded according to the latest version of the 
MedDRA dictionary. Incidence tables of the number and percentage of subjects by system organ 
class (SOC) and preferred term (PT) by cohort/dose and overall will be provided for the Safety 
Population.  
 
Medical history will also be listed by subject.  
 
6.4. 
Prior and Concomitant Medication 
 
Medications will be separated into prior and concomitant medications. Prior medications are those 
with start and stop date before the date of study drug administration. Concomitant medications are 
those with either start date prior to the date of study drug administration and were ongoing while 


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for the duration of the study or those with start date after the date of study drug administration. A 
medication can be considered both prior and concomitant. 
All medications will be coded according to the latest version of the WHO Drug Dictionary and 
listed by subject. Prior and concomitant medications will be flagged accordingly.  
 
 
 
 
 


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7. 
PHARMACOKINETIC ANALYSIS 
For PK analysis of ANA001, blood samples will be collected during the entire treatment period 
according to the PK sampling schedules in the SoA. All PK analysis will be based on the PK 
Population.  
 
7.1. 
Handling Missing or Non-Quantifiable Data  
 
Below Limit of Quantitation (BLQ) values will be set to zero prior to calculation of descriptive 
statistics for the plasma concentration-time profile and the log-concentration value will be set to 
0 if the value is used in calculating geometric means. Actual elapsed time from dosing will be 
used to estimate all individual plasma pharmacokinetic parameters. Any anomalous concentration 
values observed at the pre-dose time point within a period will be examined for the understanding 
of such an occurrence, and subsequently will result in the exclusion of the subject from the 
primary AUC analysis, or be assigned a numerical zero for calculating AUC and identified in the 
CSR.  
 
BLQ value(s) that occur at the beginning of serial sampling (before the first quantifiable 
concentration value) will be treated as zero, and BLQ value(s) that occur at the end of serial 
sampling (that is, after the last quantifiable concentration value) will be treated as missing . A 
BLQ value that is embedded between two quantifiable points will be treated as missing data. If 
consecutive BLQ concentrations are followed by quantifiable concentrations in the terminal 
portion of the concentration curve, these quantifiable values will be excluded from the 
pharmacokinetic analysis by assigning them a value of missing unless otherwise warranted by the 
concentration-time profile. 
 
7.2. 
Pharmacokinetic Concentration Analysis  
 
Plasma concentrations of ANA001 will be summarized by cohort/dose and nominal timepoint with 
descriptive statistics: n, mean, SD, CV%, geometric mean, geometric CV%, median, minimum, 
and maximum. 
 
Linear and semi-logarithmic plots of the mean plasma concentration by actual sampling time will 
be provided by cohort. For the MAD part, these plots will be done for both Day 1 and Day 7. 
Individual subject plots may also be provided for ANA001 in linear and semi-logarithmic plasma 
concentrations. Additionally, for the MAD part, plots of the trough concentrations by dosing 
intervals may be plotted to understand the achievement of steady state. 
 
For each part, plasma concentrations of ANA001 at each nominal timepoint will be listed by 
subject.  
 
7.3. 
Pharmacokinetic Parameter Analysis  
 


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The following PK parameters will be calculated: 
 
• AUC(0-t) 
• AUC(0-last) 
• AUC(0-∞) (SAD only) 
• Cmax 
• tmax 
• t1/2 
• CL/F 
• Vz/F  
Actual timepoints will be used for non-compartmental analysis (NCA) calculation. 
The area under the curve (AUC) is estimated using linear/log trapezoidal rule with at least three 
measurable concentrations. For profiles including less than three consecutive measurable 
timepoints or more than two embedded BLQ values, no AUC will be reported.  
 
Analysis for the SAD Part: 
For the SAD part of the study, in calculating AUC0-∞ , the terminal elimination phase will be 
identified by regression analysis using logarithmically transformed plasma concentrations. An 
elimination phase will be considered invalid for the following situations: 
• 
It has less than 3 data points; 
• 
It has a positive slope (or correlation coefficient [r] > 0); 
Adjusted r2 ≤ 0.8. (Adjusted r2 = 1-{(1-r2)×(n-1)}/(n-2), where n is the number of data points in 
the regression and r2 is the square of the correlation coefficient.).PK parameters will be 
summarized using descriptive statistics (n, mean, SD, CV%, geometric mean, geometric CV%, 
median, minimum, and maximum) by cohort/dose. Time to maximum concentration, tmax, will be 
summarized by n, minimum, median, and maximum only. 
Dose proportionality will be assessed graphically (dose-normalized Cmax and AUC versus dose). 
If at least 3 dose groups complete the specific part of the study, dose proportionality in PK 
parameters may be evaluated by fitting the power model:  
log (Cmax or AUC) =α + β*log(dose) 
An estimate of the slope parameter β and its 90% confidence intervals will be presented. 
Individual PK parameters will be listed by subject.  
 
Analysis for the MAD Part: 
For both Day 1 and Day 7 in the MAD part of the study, PK parameters will be summarized using 
descriptive statistics (n, mean, SD, CV%, geometric mean, geometric CV%, median, minimum, 
and maximum) by cohort/dose. Time to maximum concentration, tmax, will be summarized by n, 
minimum, median, and maximum only. 


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To understand drug accumulation, geometric mean ratios, and corresponding 90% confidence 
intervals will be calculated for Day 7/Day 1 for both AUC and Cmax. 
For Day 7, dose proportionality may be assessed for steady state Cmax and AUC using methods 
similar to those employed for the SAD part of the study. 
 
 
 
 
 
 


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8. 
SAFETY ANALYSIS  
Safety analyses will be performed on the Safety Population. Safety endpoints include adverse 
events, vital signs, ECGs, physical examinations, and laboratory examinations (hematology, serum 
chemistry, and urinalysis).  
8.1. 
Adverse Events 
Adverse Events will be coded according to the latest version of the MedDRA dictionary.  
In each part of this study, surveillance of adverse events (AE) will begin from the day the subject 
signs the informed consent form (ICF) until the end of the study. 
Adverse events shall be recorded starting at signing of ICF. AEs will be defined as treatment-
emergent adverse events (TEAE), if they occurred after initiation of study treatment or if they 
started prior to study drug administration but worsened after dosing, given that worsening started 
within the treatment period. All AEs will be listed and flagged as treatment emergent or non-
treatment emergent.  
Only TEAEs shall be summarized by cohort/dose in the clinical study report. Other AEs will be 
included in the medical history and will be listed in subject listings. 
An overall summary with number and percentage of subjects with all different TEAE categories, 
like treatment-related, serious, and leading to discontinuation (MAD part only) provided by 
cohort/dose. Incidence and severity of TEAEs will be summarized by cohort/dose within System 
Organ Class (SOC), and within Preferred Term (PT).  
TEAEs will also be summarized by relationship per SOC and PT. Relationships will be categorized 
into related (includes “Related” and “Possibly Related”) and unrelated (includes “Not Related” 
and “Unlikely Related”). 
Serious TEAEs and TEAEs leading to discontinuation (MAD part only) will be summarized by 
cohort/dose with the number and percentage of subjects with treatment-emergent adverse events 
classified SOC and PT.  
All AEs will be listed by subject.   
 
8.2. 
Clinical Laboratory Assessments 
All subjects in the Safety Population will be included in the presentation of the laboratory data.   
Clinical laboratory parameters including hematology, biochemistry and urinalyses will be 
summarized by cohort/dose for each visit. In addition, changes from the baseline visit will be 
calculated for each visit if baseline and post-baseline measurements are available.  
Shift tables may also be used to display the change from before treatment to after treatment 
measurement with respect to reference ranges “missing”, “low”, “normal”, “high”, and “total”.  
All laboratory evaluations, including those that are unscheduled, will be listed by subject, and 
clinically significant values will be flagged. 


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8.3. 
Vital Signs 
All subjects in the Safety Population will be included in the presentation of vital signs data.   
Vital signs, including the assessments of systolic and diastolic blood pressure, pulse, oral 
temperature, and respiratory rate, will be summarized by cohort/dose for each visit. In addition, 
changes from the baseline visit will be calculated for each visit.  
All assessments, including those that are unscheduled, will be listed by subject, and clinically 
significant values will be flagged. 
 
8.4. 
ECG 
All subjects in the Safety Population will be included in the presentation of ECG data.  
ECG parameters, including heart rate, QT interval, QTcF interval, QTcF interval, PR interval, and 
RR interval, QRS interval, will be summarized by cohort/dose for each visit. In addition, changes 
from the baseline visit will be calculated for each visit.  
All assessments, including those that are unscheduled, will be listed by subject, and clinically 
significant findings will be flagged. 
 
8.5. 
Physical Examination 
All physical examination findings, including those that are unscheduled, will be listed by subject, 
and clinically significant findings will be flagged.  
 
 
 
 


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9. 
ANALYSIS CONVENTIONS 
Post-text tables and listings will be prepared in accordance with the current ICH Guidelines.  The 
information and explanatory notes to be provided in the “footer” or bottom of each table and listing 
will include the following information: 
 
1. Date and time of output generation; 
2. Date of database lock; 
3. SAS®1 program name, including the path that generates the output; 
4. Any other output specific details that require further elaboration. 
 
In general, tables will be formatted with a column displaying findings for all subjects.  Row entries 
in tables are made only if data exists for at least one subject (i.e., a row with all zeros will not 
appear).  The only exception to this rule applies to tables that list the termination status of subjects 
(e.g., reasons for not completing the study).  In this case, zeros will appear for study termination 
reasons that no subject satisfied.  The summary tables clearly indicate the number of subjects to 
which the data apply and unknown or not performed are distinguished from missing data. 
 
Supportive individual Subject Data Listings will be sorted and presented by cohort, subject 
number, and visit date, if applicable.     
 
Specific algorithms are discussed for imputing missing or partially missing dates, if deemed 
appropriate, under specific data topics.  Imputed or derived data are flagged in the individual 
subject data listings.  Imputed data will not be incorporated into any raw or primary datasets.  The 
imputed data will be retained in the derived / analysis datasets. Imputation for missing PK data is 
detailed in the PK section (8.1). 
The total duration for a subject on study will be calculated as the difference between the date of 
initial exposure to the study drug and the last day of observation plus one day.  All calculations for 
defining the duration on study will follow the algorithm DURATION = [STUDY COMPLETION 
OR WITHDRAW DATE – FIRST DOSING DATE + 1].   
 
This section details general conventions to be used for the statistical analyses.  The following 
conventions will be applied to all data presentations and analyses. 
 
• SAD and MAD will be displayed separately – all TLF titles will clearly indicate which part 
of the study outputs are summarizing.  
 
• Summary statistics will consist of the number and percentage of responses in each level for 
categorical variables, and the sample size (n) mean, median, standard deviation (SD), 
minimum, and maximum values for continuous variables. 
 
• All mean and median values will be formatted to one more decimal place than the measured 
value.  Standard deviation values will be formatted to two more decimal places than the 
measured value.  Minimum and maximum values will be presented with the same number 
of decimal places as the measured value. 
 


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• The number and percentage of responses will be presented in the form XX (XX.X%). 
 
• All summary tables will include the analysis set sample size (i.e., number of subjects). 
 
• Study Day 1 is defined as the first day the subject is exposed to treatment.  All study days 
are determined relative to the day of exposure to the treatment. 
  
• Baseline values will be defined as those values recorded closest to, but prior to, the first 
study treatment on Day 1. 
 
• Change from baseline will be calculated as follows:  
 
Change = Post-baseline value − baseline value. 
 
• Date variables will be formatted as DD-MMM-YYYY for presentation. 
 
• SAS Version 9.41 or higher will be the statistical software package used for all data 
analyses.   
 
• The study treatment, and subject number will be included in all data listings.  All listings 
will be sorted by subject number, visit, visit date, baseline severity, etc. as applicable.  
 
 


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10. 
REFERENCES 
1. SAS Institute Inc., SAS Version 9.4 software, Cary, NC. 
2. Certara USA, Inc. Phoenix® WinNonlin® Version 8.2 software, Princeton, NJ.  
 


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11. 
TABLES, LISTINGS, AND FIGURES  
Tables, listings and figures will be listed in a separate Data Displays Document.