Clinical trial NCT02736968
Conditions: Amoebic Dysentery Giardiasis
Keywords: Amebiasis Auranofin GI Protozoa Giardiasis Placebo-Controlled Reprofiled Drug Single-Blind
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)
Start date: 2016-11-06; Completion date: 2021-03-10
Enrollment: 93 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
This is a phase IIa, randomized, placebo-controlled, single-blinded superiority treatment study in males and non-pregnant females, 18 to 65 years of age who are in good health. This study is designed to compare placebo to once daily doses of 6mg auranofin for adults with amebiasis or giardiasis. A sample size of 68 subjects enrolled with amebiasis (34 per arm) and 68 with giardiasis (34 per arm); Power based on 60 subjects with amebiasis and 60 with giardiasis completing the study.Eligible subjects will be randomly assigned to a treatment group with auranofin (6 mg orally once daily for 5 days for giardiasis or 7 days for amebiasis) compared to a placebo group receiving similar but not identical placebo capsules. Projected duration of subject participation will be approximately 30 days of face to face visits, including the pre-enrollment screening period of up to 4 days. It is anticipated that it will take approximately 3.5 years to finish the study. Primary objectives are: 1) to compare the proportion of subjects with stools positive by rapid Enzyme Immunoassay (EIA) and positive antigen detection EIA for E. histolytica at enrollment with resolution of diarrhea (less than 3 loose stools/24 hrs) by Day 7 for E. histolytica infections. 2) to compare the proportion of subjects with stools positive by rapid EIA and positive antigen detection EIA for Giardia at enrollment with resolution of diarrhea (less than 3 loose stools/24 hrs) by Day 5 for Giardia infections.

Detailed description:
This is a phase IIa, randomized, placebo-controlled, single-blinded superiority treatment study in males and non-pregnant females, 18 to 65 years of age who are in good health. This study is designed to compare placebo to once daily doses of 6mg auranofin for adults with amebiasis or giardiasis. A sample size of 68 subjects enrolled with amebiasis (34 per arm) and 68 with giardiasis (34 per arm); Power based on 60 subjects with amebiasis and 60 with giardiasis completing the study. Eligible subjects will be randomly assigned to a treatment group with auranofin (6 mg orally once daily for 5 days for giardiasis or 7 days for amebiasis) compared to a placebo group receiving similar but not identical placebo capsules. Projected duration of subject participation will be approximately 30 days of face to face visits, including the pre-enrollment screening period of up to 4 days. It is anticipated that it will take approximately 3.5 years to finish the study. Primary objectives are: 1) to compare the proportion of subjects with stools positive by rapid Enzyme Immunoassay (EIA) and positive antigen detection EIA for E. histolytica at enrollment with resolution of diarrhea (less than 3 loose stools/24 hrs) by Day 7 for E. histolytica infections. 2) to compare the proportion of subjects with stools positive by rapid EIA and positive antigen detection EIA for Giardia at enrollment with resolution of diarrhea (less than 3 loose stools/24 hrs) by Day 5 for Giardia infections. Secondary objectives for E. histolytica infections are: 1) to compare the proportion of subjects with stools positive by rapid EIA and positive antigen detection EIA for E. histolytica and trophozoites on smear at enrollment with parasitological response (no detection of trophozoites of E. histolytica on microscopic exam) by Day 7; 2) to compare the proportion of subjects with stools positive rapid EIA and positive antigen detection EIA for E. histolytica and trophozoites on smear at enrollment with parasitological response (no detection of trophozoites on microscopic exam or negative antigen detection) by Days 3 and 5; 3) to compare the rate of decrease of trophozoites/cyst load by Quantitative Polymerase Chain Reaction (qPCR) in stools by Days 3, 5, and 7; 4) to compare the proportion of subjects with negative stool antigen tests by days 3, 5, 7, and 14; 5) to compare the proportion of subjects with sustained cure (no detection of trophozoites by microscopic exam) at 14 and 28 days; 6) to compare the proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 14 and 28 days by genotyping the initial vs. subsequent strain; 7) to compare the time to resolution of diarrhea (less than 3 loose stools/24 hours). Secondary objectives for Giardia infections are: 1) to compare the proportion of subjects with parasitological response (no detection of trophozoites on microscopic exam ) on Day 3, and 5; 2) to compare the rate of decrease of trophozoites/cyst load by qPCR in stools by Days 3 and 5; 3) to compare the proportion of subjects with negative stool antigens by days 3 and 5; 4) to compare the proportion of subjects with sustained cure (no detection of trophozoites by microscopic exam) at 14 and 28 days; 5) to compare the proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 14 and 28 days by genotyping the initial vs. subsequent strain; 6) to compare the time to resolution of diarrhea (less than 3 loose stools/24 hours).

Arms: arm 1: N=34, 6mg auranofin daily x 7 days arm 2: N=34, 6mg placebo daily x 7 days arm 3: N=34, 6mg auranofin daily x 5 days arm 4: N=34, 6mg placebo daily x 5 days
Interventions: intervention 1: Auranofin intervention 2: Placebo
Intervention descriptions: intervention 1: Auranofin is a gold-containing chemical salt available as 3mg capsules intervention 2: Placebo

Protocol documents:
Prot_000.pdf: 
________________________________________________________________________________________________________ 
 
 
Phase IIa Randomized, Single- blinded, Placebo-controlled Clinical 
Trial of the Reprofiled Drug 
Auranofin for GI Protozoa 
 
DMID Protocol Number: 15-0015 
 
 
DMID Funding Mechanism: U01AI110435-01 
 
 
Principal Investigator: Sharon Reed, MD 
 
 
DMID Program Officer: Greg Deye, MD 
 
 
DMID Medical Monitor: Mo Elsafy, MD 
 
 
Version Number: 8.0 
 
 
14 Apr 2020 
 
 
 
 
 
 


DMID Protocol 15-0015 
Version 8.0                                              14 APR 2020 
 
 
________________________________________________________________________________________________________ 
i 
Statement of Compliance 
This trial will be conducted in accordance with Good Clinical Practice (GCP) as required by the 
following: 
• United States (US) Code of Federal Regulations (CFR) applicable to clinical studies 
(45 CFR Part 46; 21 CFR Part 50, 21 CFR Part 56, and 21 CFR Part 312) 
ICH E6; 62 Federal Register 25691 (1997)  
• NIH Clinical Terms of Award 
 
All key personnel (all individuals responsible for the design and conduct of this study) have 
completed Human Subjects Protection Training. 


DMID Protocol 15-0015 
Version 8.0                                              14 APR 2020 
 
 
________________________________________________________________________________________________________ 
ii 
Signature Page 
The signature below constitutes the approval of this protocol and the attachments, and provides 
the necessary assurances that this trial will be conducted according to all stipulations of the 
protocol, including all statements regarding confidentiality, and according to local legal and 
regulatory requirements and applicable U.S. federal regulations and ICH guidelines. 
 
Site Investigator:* 
 
Signed: 
 
Date: 
 
 
Sharon Reed, MD 
 
 
 
*The protocol should be signed by the local investigator who is responsible for the study 
implementation at his/her specific site; i.e., if Investigational New Drug study, the individual 
who signs the Form FDA 1572. 
 


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iii 
TABLE OF CONTENTS 
 
  
PAGE 
STATEMENT OF COMPLIANCE ................................................................................................. I 
SIGNATURE PAGE ...................................................................................................................... II 
TABLE OF CONTENTS .............................................................................................................. III 
LIST OF FIGURES ..................................................................................................................... VII 
LIST OF ABBREVIATIONS .................................................................................................... VIII 
PROTOCOL SUMMARY .............................................................................................................. 1 
SCHEMATIC OF STUDY DESIGN: ............................................................................................ 4 
1 
KEY ROLES ........................................................................................................................... 6 
2 
BACKGROUND INFORMATION AND SCIENTIFIC RATIONALE ................................ 9 
2.1 
Background Information .................................................................................................. 9 
2.1.1 Summary of Previous Pre-clinical Studies ................................................................... 9 
2.1.2 Summary of Relevant Clinical Studies ....................................................................... 12 
2.1.3 Summary of Epidemiological Data ............................................................................ 13 
2.2 
Rationale ......................................................................................................................... 13 
2.3 
Potential Risks and Benefits ........................................................................................... 14 
2.3.1 Potential Risks ............................................................................................................ 14 
2.3.2 Known Potential Benefits ............................................................................................. 15 
3 
OBJECTIVES ........................................................................................................................ 16 
3.1 Study Objectives ................................................................................................................. 16 
3.2 
Study Outcome Measures............................................................................................... 17 
3.2.1 Primary Outcome Measures ......................................................................................... 17 
3.2.2 Secondary Outcome Measures ..................................................................................... 18 
4 
STUDY DESIGN .................................................................................................................. 19 
4.1 
Sub Studies ..................................................................................................................... 19 
5 
STUDY ENROLLMENT AND WITHDRAWAL ............................................................... 20 
5.1 
Subject Inclusion Criteria ............................................................................................... 20 
5.2 
Subject Exclusion Criteria .............................................................................................. 22 
5.3 
Treatment Assignment Procedures ................................................................................. 23 
5.3.1 Enrollment and Randomization Procedures ............................................................... 23 
5.3.2 Masking Procedures ................................................................................................... 24 
5.3.3 Reasons for Withdrawal ............................................................................................. 24 


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5.3.4 Handling of Withdrawals............................................................................................ 25 
5.3.5 Termination of Study .................................................................................................. 26 
6 
STUDY INTERVENTION/INVESTIGATIONAL PRODUCT .......................................... 27 
6.1 
Study Product Description ............................................................................................. 27 
6.1.1 Acquisition.................................................................................................................. 27 
6.1.2 Formulation, Packaging, and Labeling ....................................................................... 27 
6.2 
Dosage, Preparation and Administration of Study Intervention/Investigational Product
........................................................................................................................................ 28 
6.3 
Modification of Study Intervention/Investigational Product for a Participant ............... 29 
6.4 
Accountability Procedures for the Study Intervention/Investigational Product(s) ........ 29 
6.5 
Assessment of Subject Compliance with Study Intervention/Investigational Product(s)
........................................................................................................................................ 29 
6.6 
Concomitant Medications/Treatments ........................................................................... 29 
7 
STUDY SCHEDULE ............................................................................................................ 31 
7.1 
Screening Visits .............................................................................................................. 31 
7.1.1 Visit 00A, Day -4 to -1, Screening 1, icddr,b or Rajshahi Medical College Hospitals
 ................................................................................................................................... 31 
7.1.2 Visit 00B, Day -4 to -1, Screening 2, icddr, b or  Rajshahi Medical College Hospitals
 ................................................................................................................................... 31 
7.2 
Visit 01, Day 1, Enrollment, Dose 1, icddr,b or Rajshahi Medical College Hospital or 
Clinic .............................................................................................................................. 32 
7.3 
Follow-up Visits ............................................................................................................. 33 
7.3.1 Visit 02, Day 2 ............................................................................................................ 33 
7.3.2 Visit 03, Day 3 ............................................................................................................ 33 
7.3.3 Visit 04, Day 4 ............................................................................................................ 34 
7.3.4 Visit 05, Day 5 ............................................................................................................ 34 
7.3.5 Visit 06, Day 6 (Amebiasis arm only) ........................................................................ 34 
7.3.6 Visit 07, Day 7 (Window + 1 day) icddr,b or Rajshahi Medical College Hospitals .. 35 
7.3.7 Visit 08, Day 14 (Window: +/- 3 days) ........................................................................ 35 
7.3.8 Visit 09, Day 28 (Window: +/- 3 days) ...................................................................... 36 
7.3.9 Visit 10, Day 30 (Window: +/- 3 days) ...................................................................... 36 
7.4 
Early Termination Visit .................................................................................................. 36 
7.5 
Unscheduled Visit .......................................................................................................... 36 
7.6 
Pregnancy Visit .............................................................................................................. 37 
8 
STUDY PROCEDURES/EVALUATIONS.......................................................................... 38 
8.1 
Clinical Evaluations ....................................................................................................... 38 
8.2 Laboratory Evaluations ....................................................................................................... 38 
8.2.1 Clinical Laboratory Evaluations ................................................................................... 38 
8.2.2 Specimen Preparation, Handling and Shipping ............................................................ 40 


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v 
9 
ASSESSMENT OF SAFETY ............................................................................................... 41 
9.1 Specification of Safety Parameters ..................................................................................... 41 
9.1 
42 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety Parameters ...... 42 
9.2.1 Adverse Events ............................................................................................................. 42 
9.2.2 Solicited Events ............................................................................................................ 43 
9.2.3 Additional Adverse Event Severity Grading ................................................................ 45 
9.2.4 Serious Adverse Events ................................................................................................ 47 
9.2.5 Procedures to be Followed in the Event of Abnormal Laboratory Test Values or 
Abnormal Clinical Findings ...................................................................................... 47 
9.3 Reporting Procedures .......................................................................................................... 48 
9.3.1 Serious Adverse Events ................................................................................................ 48 
9.3.2 Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND .............. 49 
9.3.3 Reporting of Pregnancy ................................................................................................ 49 
9.4 Type and Duration of the Follow-up of Participants after Adverse Events ........................ 49 
9.4.1. Halting Rules .................................................................................................................. 50 
9.4.2. Safety Oversight (DSMB) .............................................................................................. 51 
9.4.2 Independent Safety Monitor (ISM) ............................................................................ 51 
9.4.3 Data Safety Monitoring Board (DSMB) .................................................................... 51 
10. CLINICAL MONITORING STRUCTURE ......................................................................... 53 
10.1 Site Monitoring Plan ......................................................................................................... 53 
11. STATISTICAL CONSIDERATIONS .................................................................................. 54 
11.1 Study Hypotheses .............................................................................................................. 54 
11.2 Sample Size Considerations .............................................................................................. 54 
11.3 Planned Interim Analyses .................................................................................................. 55 
11.3.1 Safety Review ............................................................................................................. 55 
11.4 Final Analysis Plan............................................................................................................ 56 
12 SOURCE DOCUMENTS AND ACCESS TO SOURCE DATA/DOCUMENTS .............. 59 
13 QUALITY CONTROL AND QUALITY ASSURANCE .................................................... 60 
14 ETHICS/PROTECTION OF HUMAN SUBJECTS ............................................................... 61 
14.1 Ethical Standard ................................................................................................................ 61 
14.2 Institutional Review Board/Ethics Committee .................................................................. 61 
14.3 Informed Consent Process ................................................................................................. 61 
14.4 Exclusion of Women, Minorities, and Children  (Special Populations) ........................... 63 
14.5 Subject Confidentiality ...................................................................................................... 63 
14.6 Study Discontinuation ....................................................................................................... 64 
14.7 Future Use of Stored Specimens ....................................................................................... 64 
15 DATA HANDLING AND RECORD KEEPING ................................................................... 65 


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vi 
15.1 Data Management Responsibilities ................................................................................... 65 
15.2 Data Capture Methods ....................................................................................................... 65 
15.3 Types of Data .................................................................................................................... 66 
15.4 Timing/Reports.................................................................................................................. 66 
15.5 Study Records Retention ................................................................................................... 66 
15.6 Protocol Deviations ........................................................................................................... 66 
16 PUBLICATION POLICY ....................................................................................................... 68 
17 SCIENTIFIC REFERENCES .................................................................................................. 70 
APPENDIX A:   SCHEDULE OF EVENTS ................................................................................. 1 
APPENDIX B:LAB PROCESSING FLOW SHEET: CLINICAL/ SAFETY LAB TESTS ......... 4 
APPENDIX C: ICDDR,B NORMAL LABORATORY VALUES (APPLICABLE FOR BOTH 
SITES) ................................................................................................................................... 6 
 
 
 


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vii 
LIST OF FIGURES 
Figure 1: Killing of metronidazole-sensitive and resistant strains by metronidazole (Mz) and 
auranofin (AF) .............................................................................................................................. 10 
Figure 2: Effect of auranofin on amebic colitis in mice. .............................................................. 10 
Figure 3: Efficacy of auranofin in rodent models of giardiasis .................................................... 11 
Figure 4: Efficacy of auranofin and metronidazole against Entamoeba cysts .............................. 12 
 
 


DMID Protocol 15-0015                          Version 8.0 
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________________________________________________________________________________________________________ 
 
 
List of Abbreviations 
AE 
Adverse Event/Adverse Experience 
CAR 
Clinical Agents Repository 
CFR 
Code of Federal Regulations 
CIOMS 
Council for International Organizations of Medical Sciences 
CONSORT 
Consolidated Standards of Reporting Trials 
CRF 
Case Report Form 
CRO 
Contract Research Organization 
DHHS 
Department of Health and Human Services 
DMID 
Division of Microbiology and Infectious Diseases, NIAID, NIH, 
DHHS 
DSMB 
Data and Safety Monitoring Board 
eCRF 
Electronic Case Report Form 
FDA 
Food and Drug Administration 
FWA 
Federalwide Assurance 
GCP 
Good Clinical Practice 
HIPAA 
Health Insurance Portability and Accountability Act 
IB 
Investigator’s Brochure 
icddr,b 
International Centre for Diarrhoeal Disease Research-
Bangladesh 
ICF 
Informed Consent Form 
ICH 
International Conference on Harmonisation 
ICMJE 
International Committee of Medical Journal Editors 
IDE 
Investigational Device Exemption 
IEC 
Independent or Institutional Ethics Committee 
IND 
Investigational New Drug Application 
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
JAMA 
Journal of the American Medical Association 
MedDRA 
Medical Dictionary for Regulatory Activities 
MOP 
Manual of Procedures 
N 
Number (typically refers to subjects) 
NCI 
National Cancer Institute, NIH, DHHS 
NDA 
New Drug Application 
NEJM 
New England Journal of Medicine 
NIAID 
National Institute of Allergy and Infectious Diseases, NIH, 
DHHS 


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________________________________________________________________________________________________________ 
 
 
NIH 
National Institutes of Health 
OCRA 
Office of Clinical Research Affairs, DMID, NIAID, NIH, 
DHHS 
OHRP 
Office for Human Research Protections 
OHSR 
Office for Human Subjects Research 
ORA 
Office of Regulatory Affairs, DMID, NIAID, NIH, DHHS 
PHI 
Protected Health Information 
PI 
Principal Investigator 
PK 
Pharmacokinetics 
QA 
Quality Assurance 
QC 
Quality Control 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SDCC 
Statistical and Data Coordinating Center 
SMC 
Safety Monitoring Committee 
SOP 
Standard Operating Procedure 
US 
United States 
WHO 
World Health Organization 
 
 


DMID Protocol 15-0015                                        Version 8.0 
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1 
Protocol Summary 
Title: 
Phase IIa Randomized, Single-blinded, Placebo-controlled 
Clinical Trial of the Reprofiled Drug, Auranofin for GI Protozoa 
Short Title: 
Auranofin for GI Protozoa 
Phase: 
IIa 
Population: 
136 symptomatic adults >18 to 65 years old from Bangladesh with 
amebiasis or giardiasis 
Number of Sites: 
1 
Study Duration:  
3.5 years 
Subject Participation 
Duration: 
7 days treatment for amebiasis and 5 days for giardiasis with follow-
up through Day 28 
Description of Agent of 
Intervention: 
Auranofin (Ridaura) 6 mg, or similar placebo 
Objectives: 
Primary E. histolytica: 
• To compare the proportion of subjects with stools positive by 
rapid EIA and positive antigen detection EIA for E. histolytica at 
enrollment with resolution of diarrhea (less than 3 loose stools/24 
hrs) by Day 7  
Secondary E. histolytica: 
• To compare the proportion of subjects with stools positive by 
rapid EIA and positive antigen detection EIA for E. histolytica 
and trophozoites on smear at enrollment with parasitological 
response (no detection of trophozoites of E. histolytica on 
microscopic exam by Days 7 
• To compare the proportion of subjects with stools positive rapid 
EIA and positive antigen detection EIA for E. histolytica and 
trophozoites on smear at enrollment with parasitological response 


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2 
(no detection of trophozoites on microscopic exam or negative 
antigen detection) by Day 3 and 5 
• To compare the rate of decrease of trophozoites/cyst load by 
qPCR in stools by Days 3, 5, and 7 
• To compare the proportion of subjects with negative stool antigen 
tests by days 3, 5, 7, and 14 
• To compare the proportion of subjects with sustained cure (no 
detection of trophozoites by microscopic exam) at 14 and 28 days  
• To compare the proportion of subjects with relapse (same strain) 
or re-infection (new strain) with positive stools at 14 and 28 days 
by genotyping the initial vs. subsequent strain 
• To compare the time to resolution of diarrhea (less than 3 loose 
stools/24 hours) 
Primary Giardia: 
• To compare the proportion of subjects with stools positive by 
rapid EIA and positive antigen detection EIA for Giardia at 
enrollment with resolution of diarrhea (less than 3 loose stools/24 
hours) by Day 5  
Secondary Giardia: 
• To compare the proportion of subjects with parasitological 
response (no detection of trophozoites on microscopic exam) on 
Days 3 and 5  
 
• To compare the rate of decrease of trophozoites/cyst load by qPCR 
in stools by Days 3 and 5 
 
• To compare the proportion of subjects with negative stool antigens 
by days 3 and 5. 
• To compare the proportion of subjects with sustained cure (no 
detection of r trophozoites by microscopic exam)  at 14 and 28 
days  


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3 
• To compare the proportion of subjects with relapse (same strain) 
or re-infection (new strain) with positive stools at 14 and 28 days 
by genotyping the initial vs. subsequent strain 
• To compare the time to resolution of diarrhea (less than 3 loose 
stools/24 hours) 
Description of Study 
Design: 
Randomized, placebo-controlled, single-blinded superiority treatment 
study to compare placebo to once daily doses of auranofin for adults 
with amebiasis or giardiasis. A sample size of 68 subjects enrolled 
with amebiasis (34 per arm) and 68 with giardiasis (34 per arm); 
Power based on 60 subjects with amebiasis and 60 with giardiasis 
completing the study 
Estimated Time to 
Complete Enrollment: 
3.5 years 


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4 
Schematic of Study Design: 
 
 
Prior to Enrollment 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
 
 
 
Day 2 
 
 
 
Day 3 
 
 
 
 
Day 4 
 
N=34 
Arm 1 
Active
Clinical Assessment, Obtain stool specimen 
Administer Study Drug 
N=68 Positive for E. histolytica 
 
N=68 Positive for G. lamblia 
Clinical and AE Assessment 
Administer Study Drug 
N=34 
Arm 1 
Active 
N=34 
Arm 2 
Placebo 
N=34 
Arm 2 
Placebo 
Clinical and AE Assessment 
Administer Study Drug 
Clinical and AE Assessment, Obtain stool specimen 
Administer Study Drug 
Obtain informed consent. Screen subjects by 
criteria. Obtain demographic/clinical data, and stool specimen at visit 00A 
and screening blood, urine, and pregnancy testing at Visit 00B.   


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5 
 
 
 
Day 5 
 
 
 
 
Day 6 
 
 
 
Day 7 
 
 
 
 
Day 14 
 
 
 
 
 
Day 28 
 
 
 
 
 
 
 
 
  
Day 30 
Clinical and AE Assessment, Obtain urine, blood 
Administer Study Drug and obtain stool for Amebiasis patients only 
Clinical and AE Assessment 
Administer Study Drug to Amebiasis patients only 
Clinical and AE Assessment 
Obtain stool specimen 
Clinical and AE Assessment, Obtain stool specimen 
Discharge from Study 
Administer standard of care treatment with metronidazole to all 
subjects who are still positive at Day 28 for Giardia or for E. 
histolytica
 
 
Clinical and AE Assessment, Obtain stool specimen 
Administer Study Drug 


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6 
1 
KEY ROLES 
Institutions: University of California, San Diego 
 
9500 Gilman Dr. 
 
La Jolla, CA 92093-0612 
 
Contact Person: Sharon Reed, MD 
 
Phone: 858-822-2808 
 
Fax: 858-534-5724 
 
Email: slreed@ucsd.edu 
 
 
 
University of Virginia School of Medicine 
 
 
Carter Harrison Building MR6, Rm. 1709 
PO Box 801340 
345 CRISPELL DR 
 
Charlottesville, VA 22908  
 
Contact Person: William Petri, Jr., MD, PhD 
 
Phone: 434-924-5621 
 
Fax: 434-924-0075 
 
Email: wap3g@virginia.edu 
 
 
International Center for Diarrheal Disease Research, Bangladesh 
 
Mohakhali, Dhaka-1212 
 
Bangladesh 
 
Contact Person: Dr. Rashidul Haque, MD, PhD 
 
Phone: 880-2-8860523 
 
Fax: 880 28812529 
 
Email: rhaque@icddrb.org 
 
 
Rajshahi Medical College Hospital 
 
Rajshahi 6000, Bangladesh 
 
Contact Person: Dr. Faisal Alam, MD 
 
Phone cell: +8801558368966 
 
Email: faisal_imm@yahoo.com 
 
 
 


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7 
 
 
INDIVIDUALS: 
 
Principal Investigator: 
 
 
 
 
Program Officer/Medical Officer: 
 
 
Sharon Reed, MD 
University of California, San Diego 
9500 Gilman Dr. 
La Jolla, CA 92093-0612 
Contact Person: Sharon Reed, MD 
Phone: 858-822-2808 
Fax: 858-534-5724 
Email: slreed@ucsd.edu 
 
Gregory A. Deye, MD 
Parasitology & International Programs Branch (HNM53) 
Division of Microbiology & Infectious Diseases 
National Institute of Allergy & Infectious Diseases 
National Institutes of Health 
BG 5601FL RM 8A39 
Mail Stop 9825 
5601 Fishers Lane 
Bethesda, MD 20852 
 
Medical Monitor: 
 
 
 
 
 
 
Mo Elsafy, MD,  
MSc Medical Monitor Office of Clinical Research Affairs  
Division of Microbiology and Infectious Diseases  
5601 Fishers Lane 7E56 MSC 9826 Bethesda, MD 20892 
Mail Stop: 9826  
Phone: 240-292-4241  
Email: elsafymm@nih.gov 
 
Clinical Project Manager: 
 
Effie Y.H. Nomicos, RN, MSN, CCRP 
NIH/NIAID/DMID 
5601 Fisher’s Lane, Room 8A35 
Bethesda, MD 20892 
Mail Stop 9825 
Phone: 240-627-3329 
BB: 240-507-0343 
Email:enomicos@mail.nih.gov 
 


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Statistical and Data Coordinating 
Center: 
 
The Emmes Corporation 
401 N. Washington St., Suite 700 
Rockville, MD 20850 
Phone: 301-251-1161 
Fax: 301-251-1355 
Email: parasitology@emmes.com 
 
Sonia Jain, PhD 
Co-Director, UCSD Biostatistics Research Center 
University of California, San Diego 
9500 Gilman Drive 
La Jolla, Ca 92093-0725 
Phone: 858-822-2388 
Email: sojain@ucsd.edu 
Regulatory Affairs Specialist: 
 
Blossom T. Smith, MS 
Office of Regulatory Affairs (HNM5G) 
National Institute of Allergy & Infectious Diseases 
National Institutes of Health 
BG 5601FL RM 7F45 
Mail Stop 9826 
5601 Fishers Lane 
Bethesda, MD 20852 
Tel: 240-627-3376 
E-mail:  blossom.smith@nih.gov 
 


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2 
BACKGROUND INFORMATION AND SCIENTIFIC 
RATIONALE 
2.1 
Background Information 
2.1.1 Summary of Previous Pre-clinical Studies 
This proposed clinical trial follows from key findings in studies supported by UO1 AI0778822 
entitled, “Novel Therapeutics for Class B Protozoa,” to design and develop highly effective 
antiparasitic agents for Class B Protozoa. The development of the first high-throughput whole 
cell screens for E. histolytica (Debnath, 2012) and G. lamblia (Gut et al., 2011) against FDA-
approved drugs and bioactive compounds (a “repurposing” screen), found that auranofin had a 
ten-fold lower IC50 (0.5M) compared with metronidazole (5.2 M) for E. histolytica and an 
equivalent IC50 for Giardia (4.0 M). Auranofin is an orally available gold-containing 
compound that has been in clinical use in the treatment of rheumatoid arthritis for 25 years.  The 
ability to reprofile an existing drug will significantly shorten the time and expense required to 
develop a drug against these important parasitic infections (Ashburn and Thor, 2004). 
 
In most organisms there are two largely independent systems to detoxify reactive oxygen species, 
one based on glutathione and the other based on thioredoxin, but E. histolytica and Giardia lack 
both glutathione reductase activity and glutathione synthetic enzymes (Fahey et al, 1984).  So the 
thioredoxin reductase is a major defense in both parasites in the prevention, intervention and 
repair of damage caused by oxidative stress (Arias et al., 2007). Auranofin inhibited recombinant 
EhTrxR with an IC50 of 400 nM (Debnath, 2012) and recombinant GiardiaTrxR with an IC50 of 
150 nM (Tejman-Yarden, 2013). In addition, inhibition of EhTrxRled to increased susceptibility 
of the trophozoites to oxidants as well as the accumulation of intracellular reactive oxide species.  
We also showed that auranofin caused the accumulation of oxidized thioredoxin in amebic 
trophozoites in vitro and in vivo. Therefore, auranofin is the first drug active against Entamoeba 
and Giardia with a clearly defined target. Based on these studies, auranofin was given Orphan 
Drug status by the FDA. 
 
Activity of auranofin against metronidazole-sensitive and resistant strains of Giardia. 
The drug susceptibility of a metronidazole sensitive, Assemblage A strains (WB (ATCC 50803) 
compared to their isogenic metronidazole resistant strains (WB-M1, WB-M2). The viability of 2 
X 103 trophozoites in 96 well plates with auranofin concentrations of 5 nM -100M and 
metronidazole concentrations of 0.1 M-2 mM was determined at 48 hours in a luminescence-
based ATP assay (Tejman-Yarden, 2013). Auranofin had equivalent efficacy with Mtz-sensitive 


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10 
and resistant strains, whereas the EC50 for Mtz-resistant strains was increased 8-40 fold (Figure 
1). 
Figure 1: Killing of metronidazole-sensitive and resistant strains by metronidazole (Mz) 
and auranofin (AF)  
 
 
The % viability of a sensitive parental strain (WB) and two resistant strains (WB-M1 and M2) 
to serial dilutions of metronidazole and auranofin are shown. 
Efficacy of auranofin in animal models of amebiasis and giardiasis. Auranofin was effective in 
an in vivo model of murine amebic colitis, in which trophozoites invade the mouse cecum 
following surgical inoculation (Debnath, 2012). Auranofin was delivered by gavage at a 
concentration of 1 mg/kg for 7 days.  The parasite burden of auranofin-treated mice by 
quantitative PCR significantly decreased compared to the control (Figure 2A) in contrast to 
metronidazole (Figure 2B), which required ten times the dose for a significant effect. 
 
 
The treatment of mice with (A) auranofin at 1 mg/kg daily dose or (B) 
metronidazole is presented as the percentage of trophozoites/ gm of tissue. 
 
The efficacy of auranofin was also tested in the suckling mouse models of giardiasis. Newborn 
C57BL/6 mice (5-7 days old) were gavaged with 107 G. lamblia trophozoites (Assemblage A 
 
Figure 2: Effect of auranofin on amebic colitis in mice.  


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strain WB or Assemblage B strain 1279, infection established over two days, and then treated 
daily with 5 mg/kg auranofin for 5 days. The number of trophozoites remaining after drug 
treatment was determined by direct counts of the small intestinal contents. Auranofin treatment 
significantly decreased the trophozoite load (Figure 3).  
 
 
 
 
 
 
 
 
 
 
Newborn (5-7 days) or adult mice were infected with trophozoites of strains 106, 1279, or GS/M 
and auranofin started 2 days afterwards for 5 days. Values represent the log number of 
trophozoites present in the small intestine. 
 
Efficacy of auranofin against cysts. We also tested the efficacy of auranofin against Entamoeba 
cysts as E. histolytica cyst are resistant to metronidazole (Upcroft, 2001). Because E.histolytica 
cannot be induced to encyst in vitro, we used E. invadens, which do. Viability was tested with a 
fluorescent ATP assay (GoLive). E. invadens cysts were resistant to metronidazole, in contrast to 
auranofin (Figure 4). We will test the in vitro efficacy of auranofin against clinical E. histolytica 
cysts both in vitro and in treated patients in this study.  
 
Figure 3: Efficacy of auranofin in rodent models of giardiasis 


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Viability was determined as the % survival compared to controls in ethanol alone.  
2.1.2 Summary of Relevant Clinical Studies 
The hypothesis is that auranofin/Ridaura will be an important drug for use against metronidazole 
resistant parasites, particularly Entamoeba histolytica, Giardia, and Trichomonas.  We have both 
in vitro and in vivo data supporting this therapeutic hypothesis. The FDA approved auranofin in 
1985 with limited PK/PD data because current sensitive techniques such as mass spectrometry 
were not available. To fill this gap, DMID performed a Phase I, single center, open label, multiple 
dose study to evaluate the PK of gold, administered as auranofin (Clinical Study Report 12-0101, 
finalized May 13, 2015). Fifteen healthy subjects were enrolled to receive the same treatment 
dose (6 mg daily for 7 days) as proposed for amebiasis in this study.  The pharmacokinetics of 
gold was evaluated by Inductively Coupled Plasma-Mass Spectroscopy (ICP-MS) in plasma (up 
to Day 126) and in feces (up to Day 42). Treatment-emergent adverse events (TEAEs) were 
monitored from Day 1 until Day 14 and serious adverse events (SAEs) until Day 126. All subjects 
(15) were included in the summary analyses for fecal gold, but two subjects were excluded (13 
final) from summary analyses of plasma gold concentration because of quantifiable Day 1 pre-
dose plasma gold concentrations higher than 5% of their Cmax.  
All subjects had quantifiable plasma gold levels throughout the 24 hour dosing period with a Tmax 
at 1.5 post dose. The plasma AUC(0-24) increased approximately six-fold from 0.971 + 0.51 to 
5.05 +1.07 g-h/mL and the Cmax three-fold from 0.102 + 0.04 to 0.312 +0.08 g/mL.  On Day 
7, 11 of 15 subjects had fecal gold concentrations of 8.80 + 7.83 mg/kg and undetectable levels 
at Day 14. The study concluded that auranofin at 6 mg per day dose for 7 days was generally safe 
Figure 4: Efficacy of auranofin and metronidazole against Entamoeba cysts 


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13 
and well tolerated with no deaths or SAEs. There were no clinically notable TEAEs or subject 
withdrawn from the study (see 2.3.1 for more details).  
2.1.3 Summary of Epidemiological Data 
Up to 10% of the world's population is infected with Entamoeba, resulting in morbidity second 
only to malaria and schistosomiasis among parasitic infections (Amoebiasis, 1997). Infection is 
initiated by the ingestion of environmentally resistant cysts in contaminated water or food, which 
release motile trophozoites in the intestine.  E. histolytica trophozoites can then invade either the 
mucosa to cause symptomatic colitis, or the bloodstream to cause distant abscesses of the liver, 
lungs, or brain (Haque et al., 2003). In endemic areas such as our clinical site in Bangladesh, 40% 
of children develop amebic colitis a year (Haque, 2006).  
Giardiasis is the most common gastrointestinal parasitic infection in the U.S. (Yoder, 2010). In 
developing countries, giardiasis is a major public health problem with infection rates in many 
regions as high as 15-20% in adults and close to 100% in children (Gilman et al., 1985), leading 
to dehydration and malnutrition. Because of its worldwide prevalence, Giardia was added to the 
WHO Neglected Disease Initiative (Savioli et al., 2006). 
2.2 
Rationale 
Entamoeba histolytica and Giardia lamblia are Class B protozoa, which cause major water- and 
foodborne outbreaks worldwide. Infection is caused by the ingestion of environmentally resistant 
cysts in water or contaminated food. Everyone in the U.S. is susceptible to these parasites, and at 
least six waterborne outbreaks of amebiasis have been documented in the U.S. and 20,000 cases 
of giardiasis reported a year.  Drug resistance is also an increasing concern with the majority of 
patients worldwide treated with imidazoles, particularly metronidazole, which causes cross-
resistance to the newest approved drugs, tinidazole and nitazoxanide. Metronidazole has frequent 
side effects, is carcinogenic and mutagenic in animals, and does not kill cysts. Therefore, shorter, 
more tolerable, and cyst killing therapy is needed.  
 
This proposed clinical trial will test the hypothesis that oral auranofin is effective for the 
treatment of amebiasis and giardiasis and follows from studies supported by UO1 AI0778822 
entitled, “Novel Therapeutics for Class B Protozoa.” The overall aim of the project was to 
design and develop highly effective antiparasitic agents for Class B Protozoa. As part of whole-
cell screening of amebic trophozoites against FDA-approved drugs, one drug, auranofin, was 
found to have an IC50 of 0.5M vs. the standard drug, metronidazole (IC50= 5.2 M). We 
found that auranofin targets the E. histolytica thioredoxin reductase (EhTrxR) in vitro and is 
efficacious against amebic colitis and liver abscess in rodent models. Studies with Giardia 
demonstrated that auranofin was very effective against both metronidazole-sensitive and 
resistant strains of Giardia in vitro and in rodent models. Auranofin is an orally available gold-


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containing compound, FDA-approved in 1985 for rheumatoid arthritis, which has received 
Orphan Drug status for the treatment of amebiasis. During the planning stage of this trial, 
funded by R34 Grant AI098633-01, we have identified the International Centre for Diarrheal 
Diseases in Bangladesh (icddr,b) as the ideal clinical site under the direction of Dr. Rashidul 
Haque with Dr. William Petri as Co-PI. Potential patients are routinely screened for both E. 
histolytica and G. lamblia, so we can efficiently enroll patients infected with either parasite, 
supporting potential approval for two indications for auranofin in a single trial. This is an NIH 
Priority as the use of auranofin for amebiasis and giardiasis has been awarded both an R34 
Planning Grant as well as a Phase I Clinical Trial Grant. 
Because this trial will re-profile an FDA-approved drug that is still in use, we propose a Phase 
IIa, randomized, single-blinded, placebo-controlled superiority clinical trial comparing auranofin 
(standard dose of 6 mg daily) for 7 days for treatment of amebiasis or 5 days for giardiasis to 
placebo in adults. The two primary outcomes will be the proportion of patients with resolution of 
amebiasis by day 7 of therapy and/or resolution of giardiasis by day 5.  Any patients who remains 
asymptomatic but stool positive at 28 days will receive standard of care therapy with 
metronidazole. 
2.3 
Potential Risks and Benefits 
2.3.1 Potential Risks 
Auranofin is contraindicated in patients with a gold allergy, and not recommended during 
pregnancy or severe hepatic or renal insufficiency. The only reported drug interaction is a single 
patient with elevated phenytoin blood levels. The complications listed in the package insert for 
long term (> 1 yr) auranofin therapy include dermatologic: rash in 26%; gastrointestinal: loose 
stools (42%), abdominal pain (14%), nausea (10%); hematologic: anemia, leucopenia, 
thrombocytopenia in up to 3%; hepatic: elevated liver enzymes: 2%; mucous membranes: 
stomatitis (13%); renal: proteinuria (1%).  
In the DMID-sponsored Phase I Clinical Trial 12-0101 (Section 2.1.2), the safety of auranofin 
was assessed in 15 healthy volunteers based on adverse events collected from the first dose on 
Day 1 through Day 14 and through Day 126 for SAEs. Four subjects (26.7%) reported Treatment-
related TEAEs, one with headache, one with headache and nausea, one with abdominal 
discomfort and flatulence, and one with diarrhea. All were assessed as mild by the investigator. 
There were no clinically meaningful trends or changes in clinical laboratory results, vital signs, 
or physical examination data during the study.  
Auranofin is a Category C drug whose use is not recommended in pregnant women because of 
studies in pregnant rabbits and rats with decreased maternal and fetal weight and litter size with 


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4-50X the human daily dose. The risk to pregnant women in our trial will be mitigated by careful 
screening, pregnancy testing and providing contraception (hormonal oral or injectable 
contraceptives for women in addition to providing condoms for their partners) for the length of 
the trial (5-7 days) and for three half-lives of the drug (total 4 months). Birth control method and 
compliance will be assessed during screening and throughout the subject’s participation. 
2.3.2 Known Potential Benefits 
Treatment worldwide for giardiasis relies primarily on imidazoles, particularly metronidazole, 
which was first introduced in 1959. Metronidazole has not been approved for use against Giardia 
in the U.S. and has been associated with treatment failures up to 20% (Upcroft 2001). It also 
causes frequent side effects, including a disulfiram-like effect (severe nausea and vomiting with 
alcohol ingestion), central and peripheral nervous system abnormalities (seizures, encephalopathy 
and peripheral neuropathy), and interactions with drugs metabolized by the CYP2C9 enzyme of 
the P450 system (Freeman et al., 1997). Metronidazole is also carcinogenic and teratogenic in 
animals (Bendesky et al., 2002). Nitazoxanide, a thiazolide drug, is the first drug with efficacy 
against Cryptosporidium as well as Giardia compared to placebo (Rossignol et al., 2001). 
Albendazole therapy of giardiasis has been evaluated in several clinical trials (reviewed in 
Solaymani-Mohammadi et al. 2010.) Quinacrine and Furazolidone also have activity, but are no 
longer available in the U.S. (Gardner and Hill, 2001). Metronidazole resistance is now a major 
concern as clinical failures have been linked to metronidazole-resistant Giardia (Upcroft et al., 
2001; Lemee et al., 2000), and highly resistant strains are readily produced in the laboratory 
(Dunn et al., 2010). Unfortunately, metronidazole-resistant Giardia are cross- resistant to 
nitazoxanide (Wright et al., 2003) and all metronidazole derivatives. 
In addition, auranofin may prove to be a future broad spectrum antiparasitic drug as in vitro and/or 
in vivo efficacy has already been demonstrated against amebiasis (Debnath, 2012), trichomoniasis 
(Dr. Kirk Land, University of the Pacific, personal communication), Cryptosporidium (Dr. 
Momar Ndao, personal communication), Toxoplasma gondii (Andrade, 2014), T. brucei 
(Lobanov, 2006), Leishmania infantum (Ilari, 2011), filaria (Dr. James McKerrow, UCSF, 
personal communication), and schistosomiasis (Kuntz, 2007; Angelucci, 2009). 


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3 
OBJECTIVES 
3.1 Study Objectives 
Auranofin is a “repurposed” drug with established pharmacology and is approved for human use 
by the FDA.  We will assess clinical efficacy for two new indications, amebiasis and giardiasis, 
with a randomized Phase IIa, placebo-controlled, single-blinded, superiority trial comparing 
auranofin for treatment of amebiasis and giardiasis to placebo in adults. Resolution of diarrhea 
and clearance of parasites will be the endpoints, and all subjects who remain positive at Day 28 
for giardiasis or amebiasis will receive standard of care treatment with metronidazole. Ultimately, 
the goal will be to expand the trial to include symptomatic children after proving efficacy and 
safety in adults. 
 
All subjects will have the same screening protocol, but the trial will evaluate subjects with 
amebiasis or giardiasis with separate primary and secondary endpoints.  
 
Asymptomatic subjects enrolled under prior versions of the protocol will be analyzed as per 
the corresponding objectives and study SAP.  
 
Primary E. histolytica: 
To compare the proportion of subjects with stools positive by rapid EIA and positive antigen 
detection EIA for E. histolytica at enrollment with resolution of diarrhea (less than 3 loose stools/ 
24 hours) by Day 7.  
Secondary E. histolytica: 
 
• To compare the proportion of subjects with positive rapid EIA and positive antigen 
detection EIA for E. histolytica and trophozoites on smear (wet mount or concentrated 
trichrome) at enrollment with parasitological response (no detection of trophozoites on 
microscopic exam by Day 7.  
• To compare the proportion of subjects with positive rapid EIA and positive antigen 
detection EIA for E. histolytica and trophozoites on smear at enrollment with 
parasitological response (no detection of trophozoites on microscopic exam) by Day 3 
and 5. 
• To compare the rate of decrease in trophozoite/cyst load by qPCR in stools by Days 3, 5, 
and 7. 
• To compare the proportion of subjects with negative stool antigen test by Days 3, 5, 7, 
and 14. 


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• To compare the proportion of subjects with sustained cure (no detection of trophozoites 
by microscopic exam) at 14 and 28 days. 
• To compare the proportion of subjects with relapse (same strain) or re-infection (new 
strain) with positive stools at 14 and/or 28 days by genotyping the initial vs. subsequent 
strain. 
• To compare the time to resolution of diarrhea (less than 3 loose stools/24 hours) 
 
 
Primary Giardia: 
 
To compare the proportion of subjects with stool positive rapid EIA and positive antigen detection 
EIA for Giardia at enrollment with resolution of diarrhea (less than 3 loose stools/24 hours) by 
Day 5. 
Secondary Giardia: 
• To compare the proportion of subjects with parasitological response (no 
detection of trophozoites on microscopic exam) by Day 3 and 5. 
• To compare the rate of decrease in trophozoite/cyst load by qPCR in stools by 
Days 3 and 5. 
• To compare the proportion of subjects with negative stool antigens by days 3 
and 5. 
• To compare the proportion of subjects with sustained cure (no detection of 
trophozoites by microscopic exam) at 14 and 28 days. 
• To compare the proportion of subjects with relapse (same strain) or re-
infection (new strain) with positive stools at 14 and/or 28 days by genotyping 
the initial vs. subsequent strain. 
• To compare the time to resolution of diarrhea (less than 3 loose stools/24 
hours) 
 
3.2 
Study Outcome Measures 
3.2.1 Primary Outcome Measures 
1. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for E. 
histolytica and resolution of diarrhea (less than 3 loose stools/ 24 hours) by Day 7.  
2. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for 
Giardia and resolution of diarrhea (less than 3 loose stools/24 hours) by Day 5.  


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3.2.2 Secondary Outcome Measures 
1. E. histolytica: 
a. Proportion of subjects with positive rapid EIA and positive antigen 
detection EIA for E. histolytica and trophozoites on smear at enrollment 
with parasitological response (no detection of trophozoites on microscopic 
exam) by Day 7. 
b. Proportion of subjects with positive rapid EIA and positive antigen 
detection EIA for E. histolytica and trophozoites on smear at enrollment 
with parasitological response (no detection of trophozoites on microscopic 
exam) by Day 3 and 5. 
c. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3, 5, 
and 7 
d. Proportion of subjects with negative stool antigen test by Days 3, 5, 7, and 
14 
e. Proportion of subjects with sustained cure (no detection of trophozoites by 
microscopic exam) at 14 and 28 days  
f. 
Proportion of subjects with relapse (same strain) or re-infection (new 
strain) with positive stools at 14 and 28 days by genotyping the initial vs. 
subsequent strain 
g. 
Time to resolution of diarrhea (less than 3 loose stools/24 hours) 
 
2.  Giardia: 
a. Proportion of subjects with positive rapid EIA and positive antigen 
detection EIA for Giardia and trophozoites on smear at enrollment with 
parasitological response (no detection of trophozoites on microscopic 
exam) by Day 3 and 5  
b. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3 and 
5 
c. Proportion of subjects with negative stool antigens by days 3 and 5.  
d. Proportion of subjects with sustained cure (no detection of trophozoites by 
microscopic exam) at 14 and 28 days  
e. Proportion of subjects with relapse (same strain) or re-infection (new 
strain) with positive stools at 14 and 28 days by genotyping the initial vs. 
subsequent strain 
f. Time to resolution of diarrhea (less than 3 loose stools/24 hours) 


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4 
STUDY DESIGN 
This 30-day, Phase IIa, two-arm, randomized, placebo-controlled, single-blinded, superiority 
treatment study in 136 males and non-pregnant females ≥ 18 to 65 years of age will compare 
placebo to once daily doses of auranofin (Ridaura) for adults with amebiasis (68 subjects)  or 
giardiasis (68 subjects).Potential Bangladeshi adult subjects will be identified by a health 
professional from otherwise stable patients presenting to the icddr,b or Rajshahi Medical College 
Hospitals with diarrhea (thought to be due to amebiasis or giardiasis). Rajshahi Medical College 
Hospital is being added as a new enrollment site because of their higher prevalence of amebic 
infections (Alam et al., 2014). Eligible subjects will be randomly assigned to a treatment group 
with auranofin (6 mg orally once daily for 5 days for giardiasis or 7 days for amebiasis) compared 
to a placebo group receiving similar but not identical placebo capsules. Projected duration of 
subject participation will be approximately 30 days of face to face visits, including the pre-
enrollment screening period of up to 4 days. It is anticipated that it will take approximately 3.5 
years to finish the study.   
Primary and secondary outcomes to be measured during the course of the study are outlined in 
Section 3.2. 
For additional details on study procedures and evaluations by study schedule and study visits see 
Section 7, 8, and Appendix A. 
4.1 
Sub Studies 
No sub studies are planned. 


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5 
STUDY ENROLLMENT AND WITHDRAWAL 
One hundred thirty six males and non-pregnant females, aged 18-65 years, with amebiasis or 
giardiasis who are otherwise healthy and meet all eligibility criteria, will be chosen from adult 
patients presenting to the iccdr,b  or Rajshahi Medical College Hospitals with mild diarrhea. 
 
Subject Inclusion and Exclusion Criteria must be assessed by a study clinician licensed to make 
medical diagnoses and listed on the Form FDA 1572 as the site principal investigator or sub-
investigator. 
 
No exemptions are granted on Subject Inclusion/Exclusion Criteria in DMID-sponsored studies.  
Questions about eligibility should be directed toward the DMID Medical Officer. 
5.1 
Subject Inclusion Criteria 
Subjects eligible to participate in this study must meet all of the following inclusion criteria:  
1. Provide written informed consent prior to initiation of any study procedures. 
 
2. Able to understand and comply with planned study procedures and be available for all 
study visits. 
 
3. Male or non-pregnant, non-lactating females 18-65 years of age, inclusive. Females of 
reproductive potential currently using effective contraceptive methods are eligible. 
 
4. Amebiasis or giardiasis identified by rapid EIA and positive antigen detection EIA of 
stool1.1 
 
1If a subject is infected with both E. histolytica and Giardia, they will be enrolled in the 
E. histolytica study arm. Once the Entamoeba study arm is fully enrolled, any subsequent 
dual infected subjects will be enrolled in the Giardia arm.  If a subject is infected with 
both Giardia and Cryptosporidium, they will not be enrolled.  
 
5. Has diarrhea (defined as three or more loose stools) in the past 24 hrs, but is assessed to 
be clinically stable and in otherwise good health2. 
2As determined by medical history and targeted physical examination, if indicated based 
on medical history, to evaluate acute or currently ongoing chronic medical diagnoses or 
conditions that would affect the assessment of eligibility and safety of subjects. Existing 


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medical diagnoses or conditions (except those in the Subject Exclusion Criteria) must be 
deemed as stable chronic medical conditions. A stable chronic medical condition is 
defined as no change in prescription medication, dose, or frequency of medication in the 
last 3 months (90 days) and health outcomes of the specific disease are considered to be 
within acceptable limits in the last 6 months (180 days).  Any change due to change of 
health care provider, insurance company, or that is done for financial reasons, as long as 
in the same class of medication, will not be considered a violation of this inclusion 
criterion.  Any change in prescription medication due to improvement of a disease 
outcome, as determined by the site principal investigator or appropriate sub-investigator, 
will not be considered a violation of this inclusion criterion. Subjects may be on chronic 
or as needed (prn) medications if, in the opinion of the site principal investigator or 
appropriate sub-investigator, they pose no additional risk to subject safety. Topical, nasal, 
and inhaled medications, vitamins, and contraceptives are permitted. 
 
6. Vital signs (oral temperature, pulse, and blood pressure) are all within normal protocol-
defined ranges (abnormal criteria defined in Section 9.2.3 Additional Adverse Event 
Severity Grading). 
 
7. Laboratory tests (blood urea nitrogen, creatinine, AST, ALT, white blood cells, platelets, 
hemoglobin) are all within protocol-defined ranges. 
 
Subjects will be eligible for enrollment with the following laboratory values:  
• Blood urea nitrogen less than or equal to 30 mg/dL 
• Creatinine less than or equal to 133umol/L 
• AST or ALT less than or equal to 70.0 U/L 
• White cell count between 3.5 and 13.0 inclusive(109/L) 
• Platelets between 131 and 550 (109/L) inclusive 
• Hemoglobin between 11.0 and 18.0 gm/dL inclusive 
 
8. Urinalysis with no greater than trace protein. If a high protein is confirmed to be due to 
menstruation, it should be repeated. 
 
9. Women of reproductive potential4 must have a negative urine pregnancy test within 72 
hours of starting study medications.  
 
4Female subjects who are surgically sterile via tubal sterilization, bilateral oophorectomy 
or hysterectomy, who have been postmenopausal for greater than 1 year are not considered 
to be of reproductive potential. 


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10. Female subjects participating in sexual activity that could lead to pregnancy must be using 
and continue to use highly effective5 contraception for a total of 4 months after enrollment.   
 
5Highly effective methods of contraception are defined as having low failure rates (i.e. 
less than 1% per year) when used consistently and correctly and may include, but are not 
limited to, abstinence from intercourse, monogamous relationship with a vasectomized 
partner, male condoms with spermicide, diaphragm with spermicide, intrauterine devices, 
and licensed hormonal methods.  Females on effective forms of birth control will continue 
while on the study and for the follow-up period of 4 months total. The method and 
compliance of birth control used will be confirmed and documented at all study visits. 
5.2 
Subject Exclusion Criteria 
Subjects meeting any of the following exclusion criteria at baseline will be excluded from study 
participation within 1 week of the initial visit: 
1. Known intolerance of auranofin or gold compounds. 
 
2. Pregnant or breastfeeding women or women of reproductive potential not using effective 
contraception or who plan to become pregnant or breastfeed at any given time during the 
study or within 3 months of study completion. 
 
3. Use of metronidazole within the past 7 days. 
 
4. Has any condition that would, in the opinion of the site investigator, place the subject at an 
unacceptable risk of injury or render the subject unable to meet the requirements of the 
protocol. 
 
5. Concurrent participation in other investigational protocols or receipt of an investigational 
product within the previous 30 days. 
 
6. History of alcohol or drug abuse within the last five years. 
 


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5.3 
Treatment Assignment Procedures 
5.3.1 Enrollment and Randomization Procedures 
Per International Conference on Harmonisation (ICH) guideline E6: Good Clinical Practice 
(GCP), screening records will be kept at the icddr,b or Rajshahi Medical College hospital  site to 
document the reason why an individual was screened, but failed trial entry criteria.  The reasons 
why individuals failed screening will be recorded in the Statistical and Data Coordinating 
Center’s (SDCC) Advantage EDCSM (Electronic Data Capture System). No subject may be 
screened more than twice due to a screening failure result.  
 
Randomization will be stratified by site to prevent confounding that could arise from imbalance 
in treatment allocations between sites.  Because the relative pace of enrollment between the two 
sites is unknown, the number of participants to be enrolled at each site is not fixed in advance.  
With this approach, the faster enrolling site may enroll more subjects, allowing enrollment to 
complete as quickly as possible.  Each site may enroll up to 68 subjects into each strata, so it is 
possible that one strata could be filled entirely with subjects from a single site.  With the flexible 
approach, treatment assignments may not be perfectly balanced (overall or within each site), but 
they will be approximately balanced.  With sample sizes of 68 in each strata, the impact of a small 
imbalance on power is negligible.   
Once eligibility and consent are confirmed, subjects will be stratified by site and infection (a 2-
level variable of either amebiasis vs. giardiasis), and then within each stratum, randomized 1:1 to 
receive auranofin or placebo on Day 1. Both disease groups will enroll concurrently.  Enrollment 
will continue in the other arm after one arm reaches full enrollment. All subjects will receive 
doses of the assigned medication or placebo delivered orally on Days 1, 2, 3, 4, 5, 6, and 7 for 
amebiasis and Days 1, 2, 3, 4, and 5 for giardiasis. 
The randomization code will be prepared by statisticians at the SDCC and included in the 
enrollment module for the trial. SDCC staff will generate two randomization lists and send 
corresponding documentation to the study site. The randomization lists will include the 
randomization codes that will be used to serially assign study treatment to subjects as they are 
assigned to the study (i.e., randomize subjects to study arm by study identification number).  This 
table will not contain unblinded information; study staff will not be able to see the study arm 
assignment for each study identification number.  
Eligible study subjects will be randomized under single-blind conditions by study staff members 
after consent. Since the placebo is similar but not identical to the study medication, only the lab 


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personnel are truly blind, however the participant will not be informed whether they receive study 
product or placebo. The randomization number will be recorded on the study CRFs. 
A designated individual at icddr,b and Rajshahi Medical College hospital  will be provided with 
the treatment key, which links the treatment code to the actual treatment assignment, which will 
be kept in a secure place.  
5.3.2 Masking Procedures 
This is a single-blind study because the placebo is similar, but not identical to the study drug. The 
laboratory personnel performing evaluations will be blinded to treatment assignment. 
Subjects, investigators, and study personnel performing any study-related assessments following 
study product administration will not be informed of group allocation, but could detect differences 
in study and placebo capsules. They will be instructed to perform their clinical or AE assessments 
as though blinded to prevent potential bias.  
The randomization scheme will be generated by the SDCC and provided to unblinded study 
personnel (i.e., study pharmacist).  
The Data and Safety Monitoring Board (DSMB) may receive data in aggregate and presented by 
treatment group.  The DSMB may also be provided with expected and observed rates of the 
expected AEs in an unblinded fashion.  The DSMB may be unblinded to individual study 
treatment assignments, as needed, to adequately assess safety issues. Refer to the MOP for 
unblinding procedures. 
5.3.3 Reasons for Withdrawal 
Subjects may voluntarily withdraw their consent for study participation at any time and for any 
reason, without penalty. Subjects who have received study product, regardless of the number of 
doses received, or who developed an AE or SAE will be encouraged to remain in the study to be 
followed for safety purposes. 
A subject may withdraw or be withdrawn from this study for any of the following reasons: 
• Medical disease or condition, or any new clinical findings for which continued 
participation, in the opinion of the site principal investigator or appropriate sub-
investigator, would compromise the safety of the subject, or would interfere with the 
subject's successful completion of the study, or would interfere with the evaluation of 
responses. 


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• Subject no longer meets eligibility criteria. 
• As deemed necessary by the site principal investigator or appropriate sub-investigator for 
noncompliance or other reasons. 
• Subject withdrawal of consent. 
• Subject lost to follow-up. 
• Termination of the study. 
• New information becomes available that makes further participation unsafe. 
A subject will be discontinued from further receipt of auranofin or placebo if: 
• The subject misses more than one dose of drug or placebo.  
• Any clinical adverse event, laboratory abnormality, intercurrent illness, other medical 
condition or situation occurs such that continued participation in the study would not be 
in the best interest of the subject. 
5.3.4 
Handling of Withdrawals 
The primary reason for withdrawal from the study will be recorded on the Study Status data 
collection form.  Subjects will be encouraged to complete the Early Termination Visit.  The Early 
Termination Visit procedures are listed in Section 7.4.  Although subjects are free to withdraw at 
any time or may be withdrawn by the site principal investigator or appropriate sub-investigator at 
any time, subjects who receive at least one dose of study product will be encouraged to remain in 
the study for follow-up safety assessments and collection of stool specimens.  Every attempt will 
be made to follow all adverse events, including systemic reactions, serious adverse events, and 
new-onset chronic medical conditions ongoing at the time of early withdrawal to resolution. 
In the case of subjects who fail to appear for a follow-up safety assessment, extensive effort (i.e., 
three documented contact attempts via phone calls and/or in person visits to the household of the 
participants) will be made to locate or recall them, or at least to determine their health status. 
These efforts will be documented in the subject’s records. 
Subjects who sign the informed consent form and are randomized but do not receive study product 
may be replaced with new subjects who will receive the same treatment assignment. .  Subjects 
who sign the informed consent form, are randomized and receive study product, and subsequently 


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withdraw, or are withdrawn or terminated from this study, or are lost to follow-up will not be 
replaced.   
5.3.5 Termination of Study 
Although the study sponsor has every intention of completing the study, it reserves the right to 
terminate the study at any time for clinical or administrative reasons.  Reasons for termination 
include, but are not limited to, study closure due to DSMB review and recommendation and at 
the discretion of DMID. 
 


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6 
STUDY INTERVENTION/INVESTIGATIONAL PRODUCT 
6.1 
Study Product Description 
Auranofin (2,3,4,6-tetra-O-acetyl-1-thio-D-glucopyranosato-S) (triethyl-phosphine gold), is an 
orally available gold-containing compound with established pharmacology and is approved for 
human use by the FDA. It is available in an oral form as capsules called Ridaura, which contains 
3 mg of auranofin. Auranofin 3 mg capsules are manufactured by Prometheus Laboratories, San 
Diego, CA. 
6.1.1 Acquisition 
Auranofin 3mg capsules will be purchased from a commercial supplier by Fisher BioServices. 
The placebo will be manufactured by commercial supplier through contract with Fisher 
BioServices. 
Upon request by DMID, Auranofin and placebo will be shipped to the following address: 
DMID-Clinical Material Services (CMS)  
Fisher BioServices 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 
Phone: 240-477-1350 
Fax: 240-477-1360 
Email: DMID.CMS@thermofisher.com  
Auranofin and placebo will be stored at the DMID CMS- Fisher BioServices and will be provided 
to the clinical research site upon request and with prior approval from DMID. 
6.1.2 Formulation, Packaging, and Labeling 
6.1.2.1 
 
Auranofin Capsules 
Each capsule of auranofin has an opaque brown cap and opaque tan body, contains 3 mg of white 
crystalline powder, and is imprinted with the product name RIDAURA.2 Inactive ingredients 
consist of benzyl alcohol, cellulose, cetylpyridinium chloride, D&C Red No. 33, FD&C Blue No. 
1, FD&C Red No. 40, FD&C Yellow No. 6, gelatin, lactose, magnesium stearate, povidone, 
sodium lauryl sulfate, sodium starch glycolate, starch, titanium dioxide and trace amounts of other 
inactive ingredients. Auranofin 3 mg capsules are packaged in bottles of 60 count. 
 


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6.1.2.2 
 
Placebo Capsules 
The placebo capsules will be similar to auranofin capsules with opaque brown caps and an opaque 
tan body, but will not be imprinted with the product name RIDAURA. The placebo capsules will 
be manufactured by a contract supplier through Fisher BioServices. Measures will be taken to 
mitigate potential bias by the single-blinded nature of the trial. The specific measures will be 
detailed in the MOP.  
 
6.1.2.3 
Packaging and Labeling 
The daily dose of placebo and Auranofin capsules will be identical in a tight, light-resistant 
container. The study products will be labeled to include FDA cautionary information: “Caution- 
New drug -Limited by Federal (or United States) Law to Investigational Use Only.”    
 
6.1.2.4 
Storage and Stability 
The study drugs will be sent first to icddr,b and then transported by icddr,b staff under 
continuously monitored, temperature controlled conditions to Rajshahi Medical College hospital 
for storage and dispensing as detailed in the MOP. Auranofin powder darkens slightly when 
exposed to strong light and also to some extent when stored at a temperature of 60°C or warmer; 
the darkening indicates a small degree of chemical degradation, but the effect on biologic activity 
is not known. The Auranofin capsules and the placebo will be stored between 15° and 30°C (59° 
and 86°F).  
6.2 
Dosage, Preparation and Administration of Study 
Intervention/Investigational Product 
Auranofin 6mg (administered orally as two-3mg capsules) or similar placebo (administered orally 
as two capsules) once daily for 5 days for giardiasis or 7 days for amebiasis. All shipped study 
products will be initially stored at icddr,b hospital. Study products will subsequently be 
transported to Rajshahi Hospital for storage and dispensing. 
Enrollment may occur any day of the week. The complete course of study capsules (7 days for 
amebiasis and 5 days for giardiasis) will be dispensed in individual daily containers upon 
randomization by the icddr,b or Rajshahi pharmacy and dispensed as follows: 
For subjects enrolled at the icddr,b and Rajshahi Medical College Hospitals, the first doses will 
be dispensed until the patient is discharged, with all doses thereafter managed through home visits 
or at  icddr,b or Rajshahi Medical College Hospitals or local site clinic. Refer to the protocol-
specific MOP for additional information regarding dispensing procedures. 


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For all subjects: Friday is a holiday in Bangladesh so two days’ supply will be given on Thursday 
and the Friday dose will be self-administered as instructed and documented on the next visit.  
6.3 
Modification of Study Intervention/Investigational Product for a 
Participant 
Because auranofin is an FDA-approved drug, which has undergone extensive short term toxicity 
studies in a Phase I trial, there will be no dose modification of the drug, and subjects will be 
withdrawn if they miss more than one dose. If at baseline, a subject has any exclusionary 
abnormality in the screening blood and urine tests, they will be notified and not enrolled. The 
next safety lab testing time will be at Day 7 after completion of the study agent. Any laboratory 
abnormalities at that point will be evaluated as an AE, and may be reported to the DSMB as 
requested, and followed accordingly.  
6.4 
Accountability Procedures for the Study Intervention/Investigational 
Product(s) 
After receipt of the study product, the site Principal investigator (PI) is responsible for distribution 
and disposition of these study products, and has ultimate responsibility for drug accountability.  
The site PI may delegate this responsibility to the site pharmacist for each site.  Study product 
will be distributed to the health care workers to be dispensed during the subject’s hospital stay 
and/or during daily subject visits at home or the Rajshahi Hospital or the local Rajshahi clinic. 
Study protocol records must be maintained and document logs of receipt, accountability, and 
storage temperature conditions at the icddr,b office or Rajshahi clinic.  These study product 
accountability and dispensing logs must be maintained in the study file. Upon completion of the 
study and after the final monitoring visit, unused study product will be retained until monitored 
and released for disposition as per the Sponsor.  
6.5 
Assessment of Subject Compliance with Study 
Intervention/Investigational Product(s) 
Subject compliance will be assessed through direct observation and pill counts. 
6.6 
Concomitant Medications/Treatments 
Whenever a concomitant medication or study agent is initiated or the dose changed, investigators 
must review the concomitant medications’ and study agents’ most recent package inserts or 
investigator's brochure to obtain the most current information on drug interactions, 


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contraindications, and precautions. Administration of any medications, therapies, or vaccines will 
be recorded on the appropriate data collection form. Concomitant medications will include all 
current medications and medications taken within the previous 30 days through the end of the 
follow-up period and will be reported in the electronic case report form (eCRF). Prescription and 
over-the-counter drugs will be included as well as herbals, vitamins, and supplements.  
Use of new medication should prompt evaluation for the presence of a new diagnosis or chronic 
medical disease or condition. 
There are no drugs that are prohibited because of their interactions with Ridaura, but current 
systemic metronidazole are an exclusion criterion. There are no medications whose dose should 
be adjusted because of this short course of Ridaura treatment. Any patients with clinically 
significant underlying renal, hepatic, or immunologic abnormalities will be excluded before 
enrollment. 


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7 
STUDY SCHEDULE 
Complete study schedule details listed by study visit are described below. Any visit may occur 
at icddr,b or  Rajshahi Medical College Hospitals or local site clinic,  or home (except those 
requiring blood drawing, Visit 00B and Day 7). Refer also to Section 8 and Appendix A: 
Schedule of Events. 
7.1 
Screening Visits 
7.1.1 Visit 00A, Day -4 to -1, Screening 1, icddr,b or Rajshahi Medical 
College Hospitals 
Potential adult subjects from Bangladesh (between the ages of 18-65) will be chosen from adult 
patients presenting to the icddr,b or  Rajshahi Medical College Hospitals with mild diarrhea. 
These potential subjects will be visited by a study team member or health professional from 
icddr,b or   Rajshahi Medical College Hospitals. Screening may take place as many as 4 days 
prior to enrollment. A 5-minute screener will be administered at the first screening visit and will 
be used to identify eligible persons who will be referred to a study team member to discuss 
inclusion in the study. Written informed consent will be obtained from potentially eligible 
subjects after the 5-minute screener and prior to conducting any further screening or study 
procedures. A single ICF will be used to document the volunteer’s consent to further screening 
and participation in the study.  
For subjects who are potentially eligible and who have provided informed consent, the following 
procedures will be conducted at the first screening visit:  
• Full eligibility criteria will be reviewed with subjects.   
• Demographic data, including date of birth, sex, self-reported ethnicity, and residence 
location will be collected. 
• A stool sample will be collected for detection of Giardia or E. histolytica. 
7.1.2 Visit 00B, Day -4 to -1, Screening 2, icddr, b or  Rajshahi Medical 
College Hospitals   
Only eligible subjects who have provided informed consent and stool specimens that are positive 
for Giardia or E. histolytica will have this second screening visit for blood and urine testing. This 
visit may also occur on the same day at Visit 00A and must occur at the hospital or clinic.  


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• Medical history will be obtained by interview of subjects to assure eligibility.  
• All concomitant medications taken within 30 days prior to signing the informed consent 
form will be recorded on the appropriate data collection form and eCRF. Subjects will be 
questioned specifically about any metronidazole in the past 7 days.  
• A targeted physical examination will be performed, including vital signs (oral 
temperature, pulse (taken at the wrist or neck), blood pressure) observation for rashes or 
glossitis, and an abdominal exam.  
• A urine pregnancy test will be performed on all female subjects of childbearing potential 
and must be negative. 
• Approximately 17 mL of venous blood will be collected for CBC, ALT, AST, BUN, and 
creatinine tests. These values must be confirmed to meet the eligibility criteria as outlined 
in the Subject Inclusion Criteria (see Section 5.1) prior to randomization. 
• A urine sample will be collected to test for the presence of protein. The result must be 
confirmed to meet the eligibility criteria as outlined in the Subject Inclusion Criteria (see 
Section 5.1) prior to randomization. 
7.2 
Visit 01, Day 1, Enrollment, Dose 1, icddr,b or Rajshahi Medical 
College Hospital or Clinic 
• Enrollment and randomization of subjects 
• Eligibility criteria, including results of all clinical screening laboratory evaluations, will 
be reviewed with subjects to assure continued eligibility.  
• Targeted physical exam performed, if indicated 
• 2 capsules of study product (6mg total) or placebo will be orally taken by the subject.  
• Assessment of any new symptoms since Visit 00B before first dose is given. 
• Medical history, including any concomitant medications, reviewed.  
• Stool sample will be collected for ova and parasite testing. The window for stool 
specimens will be +1 day and the date of collection noted.   


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7.3 
Follow-up Visits 
This section summarizes the follow-up procedures conducted for each subject. Complete details 
for each Study Day are provided in Appendix A. Visit windows are specified around the target 
for each visit. Follow-up visits are scheduled in reference to dosing dates as indicated for each 
visit window. Subjects will be enrolled at the icddr,b or  Rajshahi Medical College Hospitals and 
have follow-up visits in the Hospital until their diarrhea has resolved. Subsequent follow-up visits 
may be held at the icddr,b, Rajshahi Hospital, or clinic or their home except for Visit 07 which 
must be at icddr,b or  Rajshahi Hospital or clinic because blood drawing is required. When a visit 
falls on a holiday Friday, two days’ supply of study product will be given on Thursday and the 
Friday dose will be self-administered as instructed and documented upon the next visit. Staff will 
notify the subject on Friday to take the dose and document it was taken. The window for stool 
specimens will be +1 day and the date of collection noted. For visits that fall on a Friday, the 
study team member will confirm on the next scheduled visit the diarrhea status as noted on the 
Visit Documentation for the missed visit.  This will be submitted in the Data System for study 
end-point analysis. 
7.3.1 Visit 02, Day 2 
• Targeted physical exam performed, if indicated.  
• 2 capsules of study product (6mg total) or placebo will be orally taken by the subject.  
• Assessment of any side effects or adverse events.  
• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.   
• Medical history, including any concomitant medications, reviewed.  
7.3.2 Visit 03, Day 3 
• Targeted physical exam performed, if indicated.  
• 2 capsules of study product (6mg total) or placebo will be orally taken by the subject.  
• Assessment of any side effects or adverse events.  
• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.   


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• Medical history, including any concomitant medications, reviewed.  
• Stool sample will be collected for ova and parasite testing. The window for stool 
specimens will be +1 day and the date of collection noted.   
7.3.3 Visit 04, Day 4 
• Targeted physical exam performed, if indicated.  
• 2 capsules of study product (6mg total) will be orally taken by the subject.  
• Assessment of any side effects or adverse events.  
• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.   
• Medical history, including any concomitant medications, reviewed.  
7.3.4 Visit 05, Day 5 
• Targeted physical exam performed, if indicated.  
• 2 capsules of study product (6mg total) or placebo will be orally taken by the subject.  
• Assessment of any side effects or adverse events. 
• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.    
• Medical history, including any concomitant medications, reviewed.  
• Stool sample will be collected for ova and parasite testing. The window for stool 
specimens will be +1 day and the date of collection noted.   
7.3.5 Visit 06, Day 6 (Amebiasis arm only) 
• Targeted physical exam performed, if indicated  
• 2 capsules of study product (6mg total) or placebo will be orally taken by the subject.  
• Assessment of any side effects or adverse events.  


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• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.   
• Medical history, including any concomitant medications, reviewed.  
7.3.6 Visit 07, Day 7 (Window + 1 day) icddr,b or Rajshahi Medical 
College Hospitals 
• Targeted physical exam performed, if indicated.  
• 2 capsules of study product (6mg total) or placebo will be orally taken by the subject 
(amebiasis arm only).  
• Assessment of any side effects or adverse events.  
• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.   
• Medical history, including any concomitant medications, reviewed.  
• Stool sample will be collected for ova and parasite testing (amebiasis patients only) The 
window for stool specimens will be +1 day and the date of collection noted.   
• Approximately 17 mL of venous blood will be collected for CBC, ALT, AST, BUN, and 
creatinine tests. The window for blood specimens will be +1 day and the date of collection 
noted.   
• A urine sample will be collected to test for the presence of protein.  
7.3.7 Visit 08, Day 14 (Window: +/- 3 days) 
• Targeted physical exam performed, if indicated.  
• Assessment of any side effects or adverse events. 
• Any subject with worsening diarrhea as assessed by the study physician, may have their 
dose discontinued and receive standard of care metronidazole.   
• Medical history, including any concomitant medications, reviewed.  


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• Stool sample will be collected for ova and parasite testing. The window for stool 
specimens will be +/- 3 days from the visit date and the date of collection noted.  
7.3.8  Visit 09, Day 28 (Window: +/- 3 days) 
The ninth study visit will occur on Day 28 when subjects will be assessed for medical history, 
targeted physical exam, concomitant medications review, and a final stool sample collected. 
Subjects will be instructed to contact the study staff, per the informed consent form, if any study 
product related adverse events develop in the following three months. 
7.3.9 Visit 10, Day 30 (Window: +/- 3 days)  
Visit 10 is a visit for any study patient who remains stool positive for Giardia or E. histolytica in 
the Day 28 stool specimen to receive the standard of care treatment for giardiasis or amebiasis. 
Subjects will be instructed to contact the study staff, per the informed consent form, if any adverse 
events develop in the following three months. 
7.4 
 Early Termination Visit 
 
Subjects may withdraw voluntarily from participation in the study at any time.  Subjects may also 
withdraw voluntarily from receiving the study intervention for any reason. If early termination 
occurs and if the subject is willing, a final assessment for symptoms will be performed and a stool 
specimen obtained.  
 
If voluntary withdrawal occurs, the subject will be asked to continue scheduled evaluations, 
complete an end-of-study evaluation, and be given appropriate care under medical supervision 
until the symptoms of any adverse event resolve or the subject’s condition becomes stable. 
7.5 
Unscheduled Visit 
The icddr,b and   Rajshahi Medical College Hospital Study Teams have local clinic facilities 
that are open six days a week for unscheduled visits for subjects or their families. This has 
proven very successful for subject retention as well. If the subject needs to be seen on the one 
day a week the clinic is closed, there is a doctor on call for research studies  or in Rajshahi or 
subjects also may come to the icddr,b or  Rajshahi Medical College Hospitals or local Rajshahi 
site clinic. 


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Unscheduled visits may occur at any time during the study.  Any of the following activities may 
be performed: 
• Targeted physical exam, if indicated.  
• Assessment of any side effects or adverse events. 
• Medical history, including any concomitant medications, reviewed. 
• If a subject complains of abdominal pain or diarrhea, a stool will be collected for ova 
and parasite to document whether Giardia or Entamoeba are present 
 
 
 
7.6 
Pregnancy Visit 
 
If a subject should become pregnant at any time during the study, she will be asked to stay in the 
study for continued evaluation. If she leaves the study before the end of her pregnancy, the study 
staff will request permission for her to be contacted at the end of her pregnancy so that she and 
her baby can be evaluated. 
 


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8 
STUDY PROCEDURES/EVALUATIONS 
 
8.1 
Clinical Evaluations 
Medical History: Complete medical history will be obtained by interview of the subjects at the 
screening visit (Visit 00B) and will be reviewed and updated on Day 1 (Visit 01) prior to the first 
study product administration.  Subjects will be queried regarding a history of significant medical 
disorders of the head, eyes, ears, nose, throat, mouth, cardiovascular system, lungs, 
gastrointestinal tract, liver, pancreas, kidney, urologic system, nervous system, blood, lymph 
nodes, endocrine system, musculoskeletal system, skin, and genital/reproductive tract.  A history 
of any allergies, cancer, immunodeficiency, psychiatric illness, substance abuse, and autoimmune 
disease will be solicited.  At follow-up visits after the first dose, an interim medical history will 
be obtained, when indicated, by interview of the subjects noting any changes since the previous 
visit.   
Concomitant Medications: Concomitant medications will include a review of all current 
medications and medications taken within 30 days prior to signing the informed consent form and 
through approximately 30 days after the first study product administration.  Prescription and over-
the-counter drugs will be included as well as vitamins and supplements.  Assessment of eligibility 
will also include a review of all permitted and prohibited medications per the Subject Inclusion 
and Exclusion Criteria (see Section 5). 
Physical Examination and Vital Signs: A physical examination limited to vital signs (blood 
pressure, pulse, temperature), observation for rashes or glossitis, and an abdominal exam will be 
performed at screening (Visit 00B). On Day 1 (Visit 01) prior to the first study product 
administration and at follow-up visits after the first administration of study product, a targeted 
physical examination will be performed, if indicated, based on subject’s interim medical history. 
Adverse Events:  Subjects will be interviewed to determine the occurrence of unsolicited AEs. 
8.2 Laboratory Evaluations 
8.2.1 Clinical Laboratory Evaluations 
Laboratory Evaluations will include: 
1. Stool examination: Stool samples will be obtained for ova and parasite testing.  
Subjects screened at icddr, b or Rajshahi Hospital will be given a stool cup for Day 1 


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sampling. Once discharged, all subjects will perform stool collections at icddr,b, 
Rajshahi Hospital, clinic or home on Days 3, 5, 7(window +1 day), 14 (window +3 
days) and 28 (window +3 days).The specimen will be kept in a cooler with an ice pack 
at the subject’s house until picked up by a Field Research Assistant and placed in a 
cooler with an ice pack for transport to the lab at icddr,b or  Rajshahi Medical College. 
Initial screening involves having a positive stool sample test for the E. histolytica or 
Giardia parasites. All laboratory personnel will be blinded, and the specimens de-
identified. 
a. Testing of stool samples will be performed by the icddr,b laboratory or 
Rajshahi Hospital  laboratory and will include: 
i. Rapid EIA and antigen detection EIA to detect E. histolytica and Giardia 
antigens to identify protozoal infections; 
ii. Wet mount for ova and parasite examination; 
iii. PVA fixation for concentration and trichrome staining for ova and parasite 
examination  
iv. Qualitative PCR to detect E. histolytica and Giardia 
a.  Off-site testing of stool samples collected at Rajshahi will be transported to icddr, 
b and shipped to UCSD to include quantitative E. histolytica or Giardia testing 
performed at UCSD on shipped aliquots of DNA extracted from positive stools at 
icddr,b.  De-identified non-infectious DNA specimens labeled with the subject code 
will be obtained from stools on Days 1, 3, 5, 7, 14, and 28 following enrollment. 
Quantitative PCR assays have been developed based on the small subunit rRNA gene, 
which is highly conserved among E. histolytica (Taniuchi, 2011) and Giardia isolates 
(Haque, 2007). 
2. 
Urine testing: 
a. Pregnancy testing at both hospital sites. Upon screening: as a condition of 
enrollment, all women of child bearing age must agree to have a pregnancy test, 
receive the results, and use effective birth control for the next four months.  Testing 
will be performed on urine using HCG test strip assays. If the test is positive or the 
woman is breastfeeding, they will not be enrolled, as auranofin is contraindicated in 
pregnant or breastfeeding women. Women who are enrolled in the study will continue 
their current effective birth control for 4 months after enrollment.  
b. Urine for proteinuria. On Visit 00B and 7: urine will be collected from all 
participants for proteinuria assessment via dipstick testing. Proteinuria testing will be 
performed at the icddr,b 
3. 
Blood testing: Blood will be drawn for a complete blood count, BUN, creatinine, ALT, and 
AST on Visit 00B and Day 7. Blood tests will be collected at the icddr,b Hospital or Rajshahi 
Hospital or Clinic and transported to the respective clinical laboratory. 


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4. 
E. histolytica and Giardia genotyping: Any stool specimen that is still positive for 
Entamoeba or Giardia cysts or trophozoites at Days 14 and 28 will have DNA genotyping 
of the parasite performed on de-identified specimens of extracted DNA. The genotypes from 
initial and final isolates will be compared to allow determination of the rate of relapse or re-
infection without linking to individual patients. The genotyping will be performed off-site 
at the University of Virginia. There will be no human DNA genotyping performed. 
8.2.2 Specimen Preparation, Handling and Shipping 
Appropriate blood and secretion precautions will be employed by all personnel in the drawing of 
blood and shipping and handling of all specimens for this study, as currently recommended by 
the Centers for Disease Control and Prevention and the National Institutes of Health. All 
potentially infectious specimens will only be transported locally to the laboratory using packing 
mandated in the Code of Federal Regulations, 42 CFR Part 72 and IATA guidelines.  
8.2.3 Instructions for Specimen Preparation, Handling, and Storage 
Any residual serum from safety screening, aliquots of stool, and parasite DNA extracted from the 
stool of enrolled subjects only will be stored in cryovials marked with the subject identification 
number and date of specimen collection at icddr,b or Rajshahi Hospital or Clinic following 
specific consent of the subjects. Long-term stored specimens will go to the Fisher Repository.  
Future studies may include measuring gold levels in the blood or stool. Other researchers may 
request access to these specimens for collaborative studies as well. Further instructions for 
specimen preparation, handling, and storage are included in the protocol-specific MOP as 
appropriate. There will be no human DNA collection or storage. 
8.2.4 Specimen Shipment 
Only noninfectious, parasite extracted DNA specimens will be shipped to UCSD for quantitative 
parasite loads or University of Virginia for genotyping. They can be sent at room temperature by 
standard international couriers, or designated members of the study team with completed 
specimen/Transport Receipt Logs. The residual long-term stored specimens will be shipped 
according to IATA guidelines to the Fisher Repository.  


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9 
ASSESSMENT OF SAFETY 
9.1 Specification of Safety Parameters 
Because this is an FDA approved drug that has been in use for more than 25 years, the potential 
AEs have been well documented in subjects in longer term therapy for rheumatoid arthritis. Of 
the AEs listed on the Insert, the most serious reactions would be an allergic reaction to gold, 
which could include anaphylactic reactions, necrotizing enterocolitis, pulmonary fibrosis, 
exfoliative dermatitis, bone marrow aplasia or other severe hematologic disorders.  
The adverse reactions incidences listed below are based on observations of 1) 4,784 Ridaura-
treated subjects in clinical trials (2,474 U.S., 2,310 foreign), of whom 2,729 were treated more 
than one year and 573 for more than three years; and 2) post-marketing experience. The highest 
incidence is during the first six months of treatment; however, reactions can occur after many 
months of therapy. With rare exceptions, all subjects were on concomitant nonsteroidal anti-
inflammatory therapy; some of them were also taking low dosages of corticosteroids. Reactions 
occurring in more than1% of Ridaura-treated patients: 
• Gastrointestinal: loose stools or diarrhea (47%); abdominal pain (14%); nausea with 
or without vomiting (10%); constipation; anorexia*; flatulence*; dyspepsia*; 
dysgeusia. 
• Dermatological: rash (24%); pruritus (17%); hair loss; urticaria. 
• Mucous Membrane: stomatitis (13%); conjunctivitis*; glossitis. 
• Hematological: anemia; leukopenia; thrombocytopenia; eosinophilia. 
• Renal: proteinuria*; hematuria. 
• Hepatic: elevated liver enzymes. 
*Reactions marked with an asterisk occurred in 3-9% of the patients. The other 
reactions listed occurred in 1-3%. Reactions occurring in less than 1% of Ridaura-
treated patients 
• Gastrointestinal: dysphagia; gastrointestinal bleeding†; melena†; positive stool for 
occult blood†; ulcerative enterocolitis. 
 


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42 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters 
9.2.1 Adverse Events 
Adverse Event: International Conference on Harmonisation (ICH) E6 defines an AE as any 
untoward medical occurrence in a patient or clinical investigation subject administered a 
pharmaceutical product regardless of its causal relationship to the study treatment.  An AE can 
therefore be any unfavorable and unintended sign (including an abnormal laboratory finding), 
symptom, or disease temporally associated with the use of a medicinal (investigational) product.  
The occurrence of an AE may come to the attention of study personnel during study visits and 
interviews of a study recipient presenting for medical care, or upon review by a study monitor. 
AEs not meeting the protocol-defined criteria for SAEs will be captured on the appropriate data 
collection form and electronic case report form (eCRF).  Information to be collected for 
unsolicited AEs includes event description, date of onset, licensed clinician’s assessment of 
severity and relationship to study product and alternate etiology (if not related to study product) 
(assessed only by those with the training and authority to make a diagnosis and listed on the Form 
FDA 1572 as the site principal investigator or sub-investigator), date of resolution of the event, 
seriousness and outcome.  AEs while on study will be documented appropriately regardless of 
relationship.  AEs will be followed to resolution. 
Any medical condition that is present at the time that the subject is screened will be considered 
as baseline and not reported as an AE.  However, if the severity of any pre-existing medical 
condition increases, it will be recorded as an AE. 
AEs must be graded for severity and assessed for relationship to study product (see definitions 
below).  Adverse events characterized as intermittent require documentation of onset and duration 
of each episode.  The start and stop date of each reported AE will be recorded on the appropriate 
data collection form and eCRF. 
FDA defines an AE as any untoward medical occurrence associated with the use of a drug in 
humans, whether or not considered drug related. 
Severity of Event: AEs will be assessed by a licensed study physician listed on the Form FDA 
1572 as the site principal investigator or appropriate sub-investigator using a protocol-defined 
grading system (see Section 9.2.2 and 9.2.3 ).  For events not included in the protocol-defined 
grading system, the following guidelines will be used to quantify severity: 


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43 
• Mild (Grade 1):  Events require minimal or no treatment and do not interfere with the 
subject’s daily activities. 
• Moderate (Grade 2):  Events result in a low level of inconvenience or concern with 
therapeutic measures.  Moderate events may cause some interference with functioning and 
daily activities. 
• Severe (Grade 3):  Events that prevent the subject’s usual daily activities and may require 
systemic drug therapy or other treatment.  Severe events are usually incapacitating. 
The severity of systemic and clinical laboratory adverse events will be graded according to 
Section 9.2.2 and the toxicity tables included in the Appendices to this protocol. 
Changes in the severity of any AE will be documented to allow an assessment of the duration of 
the event at each level of intensity.  When AEs are intermittent, the onset and duration of each 
episode will be documented. 
Relationship to Study Products: The study physician’s assessment of an AE's relationship to 
study product is part of the documentation process, but it is not a factor in determining what is or 
is not reported in the study.  If there is any doubt as to whether a clinical observation is an AE, 
the event should be reported.  The relationship to study product must be assessed for AEs using 
the terms:  related or not related.  In a clinical trial, the study product must always be suspect.  To 
help assess, the following guidelines are used: 
• Related – There is a reasonable possibility that the study product caused the adverse event. 
Reasonable possibility means that there is evidence to suggest a causal relationship between 
the study product and the adverse event. 
• Not Related – There is not a reasonable possibility that the administration of the study product 
caused the event. 
9.2.2 Solicited Events 
Solicited events are AEs that are known to occur with this study product. Solicited AEs will not 
be collected that are related to any prescribed metronidazole. The below Toxicity Grading Scales 
will be used to grade events. 
Adverse event data will be collected at each visit using specific questions and/or targeted physical 
examination. Based on the adverse reactions listed on the package insert, expected side effects 
include: loose stools or diarrhea, abdominal pain, nausea with or without vomiting, constipation, 
anorexia, flatulence, dyspepsia, dysgeusia, pruritus, hair loss, urticarial, stomatitis, conjunctivitis, 


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44 
glossitis, and hematuria. Diarrhea, however, will not be recorded as a side effect because all 
participants will enroll with diarrhea, and because its resolution is the endpoint of the study. 
Physical examinations will look for abdominal pain, hair loss, urticaria, rashes, stomatitis, 
conjunctivitis, and glossitis. Hematologic, renal, or hepatic laboratory abnormalities will be 
picked up on testing at Day 7. Adverse event collection will continue until Day 28 (following 
three half-lives of the drug).  
 
Event 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Loose stools or diarrhea 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Abdominal Pain 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Nausea with or without 
vomiting 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Constipation 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Anorexia 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Flatulence 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Dyspepsia 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Dysgeusia 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Pruritus 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Hair loss 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Urticaria 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Stomatitis 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Conjunctivitis 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Glossitis 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 


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Event 
Mild (Grade 1) 
Moderate (Grade 2) 
Severe (Grade 3) 
Hematuria 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
Rash 
Does not interfere with 
daily activity 
Interferes with daily 
activity 
Prevents daily activity 
(incapacitating) 
 
9.2.3 Additional Adverse Event Severity Grading 
 
Vital signs will be graded as follows: 
 
Vital Signs* 
Vital Signs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Fever (°C) ** 
 
37.8-38.4 
 
38.5-38.9 
 
>= 39.0 
 
Hypertension 
(systolic) mm Hg 
141-150 
151-155 
>155 or  
an ER visit or hospitalization 
for malignant hypertension 
Hypertension 
(diastolic) mm Hg 
91-95 
96-100 
>100 or  
an ER visit or hospitalization 
for malignant hypertension 
Hypotension 
(systolic) mm Hg 
85-89 
80-84 
<80 or  
an ER visit or hospitalization 
for hypotensive shock 
Bradycardia – beats 
per minute *** 
50-54 
or>10 bpm less than 
baseline if baseline <60 
45-49 
or>15 bpm less than 
baseline if baseline <60 
<45 
or> 20 BPM or less than 
baseline if baseline <60, 
or an ER visit or hospitalization 
for arrhythmia 
Tachycardia – beats 
per minute 
101-115 
116-130 
>130 
or an ER visit or hospitalization 
for arrhythmia 
* Subjects should be at rest for at least 15 minutes prior to vital sign measurements 
** Oral temperature; no recent hot or cold beverages or smoking 
*** When resting heart rate is between 60-100 beats per minute. Pulse and Blood Pressure assessed at Visit 1 (Day 1) will be 
considered baseline.  Sinus bradycardia among some healthy subject populations, for example, conditioned athletes maybe 
acceptable.   
 
Abnormal laboratory findings that will trigger an Adverse Event report will include: 
• 
Hematologic: WBC <3.5 or>13.0 (109/L), Hemoglobin <11.0 or >18.0.gm/dL, Platelets 
<130 or >550 (109/L) 


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• 
Hepatic: SGOT (AST) or SGPT (ALT)> 70.0 
• 
Renal: Creatinine>133 umol/L, Blood Urea Nitrogen >30 mg/dL, > 1+ proteinuria 
 
Laboratory values that are outside of the normal range, but do not meet protocol defined criteria 
as adverse events will be evaluated by a clinician for clinical significance, recorded in the 
source document, and reported as laboratory AEs if clinically significant. Other laboratory 
parameters not specified in the protocol, but performed as part of the CBC and complete 
metabolic panel, need to be evaluated by a clinician, recorded in the source document, and 
reported as laboratory AEs if clinically significant. 
 
Hematology 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
WBC 109/L (Decrease) 
3.0-3.4 
2.0-2.9 
<2.0 
WBC 109/L (Increase) 
13.1-13.9 
14.0-15.0  
>15.0  
Hgb g/dL (Decrease)  
9.1-10.9 
7.5-9.0 
<7.5 
Hgb g/dL (Increase) 
18.1-18.5 
18.6-19.0 
>19.0 
Platelets 109/L (Decrease) 
100-130 
50-99 
<50 
Platelets 109/L (Increase) 
551-600 
601-700 
>700 
 
 
 
 
 
Chemistry 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
BUN mg/dL 
31-60 
>60- and <70 
>70 
Creatinine umol/L1 (Increase)  
134-176  
177-221  
>221 
AST IU/L (Increase)  
70.1-200 
201-300 
>300 
ALT IU/L (Increase)  
70.1-200 
201-300 
>300 
 
 
Urinalysis 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Protein 
1+ 
2+ 
hospitalization or 
dialysis 
1Conversion factor from mg/dL: http://onlinelibrary.wiley.com/doi/10.1002/97808138188825.app3/pdf 
2Trace protein at screening (Visit 00B) is acceptable for inclusion into the study 


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9.2.4 Serious Adverse Events 
An adverse event or suspected adverse reaction is considered “serious” if, in the view of either 
the investigator or sponsor, it results in any of the following outcomes:  
• Death, 
• a life-threatening adverse event*,  
• inpatient hospitalization or prolongation of existing hospitalization,  
• a persistent or significant incapacity or substantial disruption of the ability to conduct normal 
life functions, or  
• a congenital anomaly/birth defect.   
• Important medical events that may not result in death, be life-threatening, or require 
hospitalizations may be considered serious when, based upon appropriate medical judgment 
they may jeopardize the patient or subject and may require medical or surgical intervention 
to prevent one of the outcomes listed in this definition.  Examples of such medical events 
include allergic bronchospasm requiring intensive treatment in an emergency room or at 
home, blood dyscrasias or convulsions that do not result in inpatient hospitalization, or the 
development of drug dependency or drug abuse. 
* Life-threatening adverse event. An adverse event is considered “life-threatening” if, in the view 
of either the investigator or sponsor, its occurrence places the patient or subject at immediate risk 
of death.  It does not include an adverse event, had it occurred in a more severe form, might have 
caused death. 
All SAEs will be: 
• recorded on the appropriate AE eCRF 
• followed through resolution by a study clinician 
• reviewed and evaluated by a study clinician 
9.2.5 Procedures to be Followed in the Event of Abnormal Laboratory 
Test Values or Abnormal Clinical Findings 
The site principal investigator or appropriate co-investigator is responsible for reporting all 
AE/SAEs that are observed or reported during the study, regardless of the relationship to study 
product.  AE/SAEs, abnormal clinical laboratory test values, or abnormal clinical findings will 


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48 
be documented, reported, and followed until levels return to baseline or until the abnormal value 
is determined to be not clinically significant. 
9.3 Reporting Procedures 
9.3.1 Serious Adverse Events 
SAEs will be followed until resolution even if this extends beyond the study-reporting period.  
Resolution of an SAE is defined as the return to pretreatment status or stabilization of the 
condition with the expectation that it will remain chronic.  
Any AE that meets a protocol-defined serious criterion must be submitted immediately 
(within 24 hours of site awareness) on an SAE form to the DMID Pharmacovigilance Group, 
at the following address:  
DMID Pharmacovigilance Group 
Clinical Research Operations and Management Support (CROMS) 
6500 Rock Spring Dr. Suite 650 
Bethesda, MD 20814, USA 
SAE Hot Line: 1-800-537-9979 (US) or 1-301-897-1709 (outside US) 
SAE FAX Phone Number: 1-800-275-7619 (US) or 1-301-897-1710 (outside US) 
SAE Email Address: PVG@dmidcroms.com 
In addition to the SAE form, selected SAE data fields must also be entered into Emmes Advantage 
eClinical system. Please see the protocol-specific MOP for details regarding this procedure. 
Other supporting documentation of the event may be requested by the DMID Pharmacovigilance 
Group and should be provided as soon as possible. 
The site will notify the ISM when an SAE is provided to the DMID Pharmacovigilance Group.  
The DMID Medical Monitor and DMID Clinical Project Manager will be notified of the SAE by 
the DMID Pharmacovigilance Group.  The DMID Medical Monitor will review and assess the 
SAE for regulatory reporting and potential impact on study subject safety and protocol conduct. 
At any time after completion of the study, if the site principal investigator or appropriate sub-
investigator becomes aware of an SAE that is suspected to be related to study product, the site 
principal investigator or appropriate sub-investigator will report the event to the DMID 
Pharmacovigilance Group. 


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9.3.2 Regulatory Reporting for Studies Conducted Under 
DMID-Sponsored IND 
Following notification from the investigator, DMID, the IND sponsor, will report any suspected 
adverse reaction that is both serious and unexpected.  DMID will report an adverse event as a 
suspected adverse reaction only if there is evidence to suggest a causal relationship between the 
drug and the adverse event.  DMID will notify FDA and all participating investigators (i.e., all 
investigators to whom the sponsor is providing drug under its INDs or under any investigator’s 
IND) in an IND safety report of potential serious risks from clinical trials or any other source, as 
soon as possible, but in no case later than 15 calendar days after the sponsor determines that the 
information qualifies for reporting as specified in 21 CFR Part 312.32. DMID will also notify 
FDA of any unexpected fatal or life-threatening suspected adverse reaction as soon as possible 
but in no case later than 7 calendar days after the sponsor’s initial receipt of the information. 
Relevant follow up information to an IND safety report will be submitted as soon as the 
information is available.  Upon request from FDA, DMID will submit to FDA any additional data 
or information that the agency deems necessary, as soon as possible, but in no case later than 15 
calendar days after receiving the request. 
All serious events designated as “not related” to study product(s), will be reported to the FDA at 
least annually in a summary format. 
9.3.3 Reporting of Pregnancy 
Pregnant women are not eligible to participate in the study. Women are counseled regarding 
prevention of pregnancy and encouraged to make every effort to avoid pregnancy during study 
participation.  If a study subject becomes pregnant during study participation, no further doses of 
study agent will be given. The basic information about the pregnancy is recorded on the 
“Pregnancy” case report form via the Emmes Advantage eClinical system. If there are 
complications during the pregnancy, the complications are recorded as adverse events in the usual 
way. The subject is asked to report outcome of the pregnancy. If there is a congenital anomaly in 
the infant, this is recorded as a serious adverse event (SAE) in the data forms for the mother (i.e., 
the study subject). Efforts will be made to follow all pregnancies reported during the course of 
the study to pregnancy outcome pending the subject’s permission. 
9.4 Type and Duration of the Follow-up of Participants after Adverse Events 
AEs and SAEs will be followed from the time of the first receipt of study drug Day 1 (Visit 01) 
through 28 days after receipt of study product. 
AEs and SAEs will be followed until resolution even if this extends beyond the study-reporting 
period (3 months after the final study visit).  Resolution of an AE/SAE is defined as the return to 


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pretreatment status or stabilization of the condition with the expectation that it will remain 
chronic. 
If the site principal investigator or appropriate sub-investigator becomes aware of a sign or 
symptom and the site principal investigator or appropriate sub-investigator decides to bring the 
subject in for an evaluation to determine etiology, then the site principal investigator or 
appropriate sub-investigator, at their own discretion, can determine what further testing is 
appropriate. 
Follow-up procedures, evaluations, and outcomes will be recorded on the appropriate data 
collection form. 
9.4.1. Halting Rules 
Individual Halting Rules: Dosing for an individual subject will be discontinued if they develop 
any grade 3 drug-related event. The DSMB has requested to be notified of any individual subject 
who experiences a grade 3 drug-related event, however, the study will continue enrollment. 
Study Halting Rules: Enrollment, dosing and study procedures will be halted for DSMB 
review/recommendation if any of the following are reported: 
• Any death occurring after administration of study drug through the subject’s last study 
visit that was not the result of trauma or accident, regardless of relatedness to study 
product. 
• Any subject experiences a study drug-related SAE from the time of receipt of study drug 
through the subject’s last study visit after relatedness is finalized by DMID. 
• Occurrence of a severe allergic/hypersensitivity reaction (anaphylaxis) within 48 hours of 
dosing, requiring hemodynamic support with vasoactive medications or mechanical 
ventilation; the signs/symptoms will include any of the following: bronchospasm, 
dyspnea, wheezing, stridor, hypoxemia urticaria, angioedema, hives, and facial or 
oropharyngeal edema. 
 
If any of the halting rules are met following any subject receipt of study drug, the study will not 
continue with the remaining enrollments without a review by and recommendation from the 
DSMB to proceed. 
DMID 
retains 
the 
authority 
to 
suspend 
additional 
enrollment 
and 
study 
interventions/administration of study product during the entire study, as applicable. 


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9.4.2. Safety Oversight (DSMB) 
9.4.2 Independent Safety Monitor (ISM) 
The ISM is a physician with relevant expertise whose primary responsibility is to provide 
independent safety monitoring in a timely fashion.  For this study an ISM is not required. 
However, at each site, and at the request of DMID, in real time, the local PI should be able to 
identify an independent physician to function as ad-hoc ISM. That person should have the 
privileges to examine the subject, review the subject medical and study record and provide an 
independent medical assessment and recommendation to DMID. 
9.4.3 Data Safety Monitoring Board (DSMB) 
Safety oversight will be conducted by a DSMB that is an independent group of experts that 
monitors subject safety and advises DMID. The DSMB members will be separate and 
independent of study personnel participating in this trial and should not have scientific, financial 
or other conflict of interest related to the study. The DSMB will consist of members with 
appropriate expertise to contribute to the interpretation of the data from this trial. The membership 
will include a chairperson and a statistician who are experienced in clinical trials conduct and 
have prior DSMB experience. The DSMB will review study progress and clinical and safety data 
at the following time points: 
• Data review for safety at study specific time frames; at least annually. 
• Data review for efficacy will be performed at the completion of enrollment. No 
interim analysis is planned.  
• Study enrollment will be stopped for DSMB review of safety data when the study 
reaches 50% enrollment (i.e. 68 subjects enrolled).  DSMB will review safety data 
after the 68th subject completed Visit 8. 
• Final review meeting: 6 to 8 months after clinical database lock to review the 
cumulative unblinded safety and efficacy data for the study.  The data will be 
provided in a standard summary format.  The DSMB may be asked to provide 
recommendations in response to questions posed by DMID. 
• Ad hoc review: may be in to an anticipated safety issue such as a halting rule being 
met. 
• Additionally there will be ongoing DSMB notification of Grade 3 drug related 
events, after which the DSMB will then decide if an Ad Hoc DSMB meeting will be 
required 
The DSMB will operate under the rules of a DMID-approved charter that will be written at the 
organizational meeting of the DSMB. At this time, each data element that the DSMB needs to 
assess will be clearly defined. Procedures for DSMB reviews/meetings will be defined in the 
charter. The DSMB will review applicable data to include, but not limited to, study progress and 
participant clinical and safety data which may include enrollment and demographic information, 


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medical history, concomitant medications, physical assessments, clinical laboratory values, 
dosing, AE/SAEs, protocol defined SAEs, and assays results. Additional data may be requested 
by the DSMB, and interim statistical reports may be generated as deemed necessary and 
appropriate by DMID. The DSMB may receive data in aggregate and presented by group. The 
DSMB will meet and review this data at scheduled time points or ad hoc as needed during the 
study as defined in the protocol and DSMB charter. As an outcome of each review/meeting, the 
DSMB will make a recommendation as to the advisability of proceeding with the study (as 
applicable), and to continue, modify, or terminate the study.  
 
DMID or the DSMB chair may convene the DSMB on an ad hoc basis according to protocol 
criteria or if there are immediate concerns regarding observations during the course of the study. 
The DMID Medical Monitor is empowered to stop enrollment and dosing of the study drug if 
halting criteria are reported. The DMID Medical Monitor will be responsible for reviewing any 
protocol defined SAEs in real time. The DSMB will review SAEs on a regular basis and ad hoc 
during the study. 
 


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10. CLINICAL MONITORING STRUCTURE 
10.1 Site Monitoring Plan 
Site monitoring is conducted to ensure that the human subject protections, study and laboratory 
procedures, study intervention administration, and data collection processes are of high quality 
and meet sponsor, ICH/GCP guidelines and applicable regulations, and that the study is conducted 
in accordance with the protocol, protocol-specific MOP and applicable sponsor standard 
operating procedures.  DMID, the sponsoring agency, or its designee will conduct site-monitoring 
visits as detailed in the clinical monitoring plan. 
Site visits will be made at standard intervals as defined by DMID and may be made more 
frequently as directed by DMID.  Monitoring visits will include, but are not limited to, review of 
regulatory files, accountability records, eCRFs, informed consent forms, medical and laboratory 
reports, and protocol and GCP compliance.  Site monitors will have access to the study site, study 
personnel, and all study documentation according to the DMID-approved site monitoring plan.  
Study monitors will meet with site principal investigators to discuss any problems and actions to 
be taken and document visit findings and discussions. 


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11. STATISTICAL CONSIDERATIONS 
11.1 Study Hypotheses 
This proposed clinical trial will test the hypothesis that oral auranofin is effective for the 
treatment of amebiasis and giardiasis, separately. Because this trial will re-profile an FDA-
approved drug that is still in use, we propose a Phase IIa, randomized, single-blinded, placebo-
controlled superiority clinical trial comparing auranofin (standard dose of 6 mg daily) for 7 days 
for treatment of amebiasis or 5 days for giardiasis to placebo in adults. The study will have two 
primary outcomes that will be tested on two different sets of 68 subjects (i.e. one set that has 
amebiasis and the other set has giardiasis).  The amebiasis primary outcome will be the 
proportion of subjects with resolution of diarrhea (less than 3 loose stools/ 24 hours) by Day 7. 
The giardiasis primary outcome will be the proportion of subjects and resolution of diarrhea (less 
than 3 loose stools/24 hours) by Day 5. Additional secondary outcomes with E. histolytica 
include the proportion of subjects with positive EIA for E. histolytica and trophozoites on smear 
at enrollment with parasitological response of amebiasis (as determined by no detection of 
trophozoites of E. histolytica on microscopic exam day 3, 5, and 7; the rate of decrease of 
trophozoite/cyst load by qPCR in stools by Days 3, 5, and 7; the proportion of subjects with a 
negative stool antigen test by Days 3, 5, 7, and 14; the proportion of subjects with sustained cure 
(no detection of trophozoites by microscopic exam) at 14 and 28 days; the proportion of subjects 
with relapse (same strain) or re-infection (new strain) with positive stools at 14 and 28 days by 
genotyping the initial vs. subsequent strain. For patients with giardiasis, secondary outcomes 
include the proportion of subjects with positive EIA for Giardia and trophozoites on smear at 
enrollment with parasitological response (no detection of trophozoites on microscopic exam) by 
Days 3 and 5; the rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3 and 5; 
the proportion of subjects with negative stool antigens by Days 3, 5, and 7; the proportion of 
subjects with sustained cure (no detection of trophozoites by microscopic exam) at 14 and 28 
days; the proportion of subjects with relapse (same strain) or re-infection (new strain) with 
positive stools at Days 14 and 28 by genotyping the initial vs. subsequent strain.  
11.2 Sample Size Considerations 
The primary amebiasis outcome for this study is to compare the proportion of subjects with 
resolution of diarrhea (less than 3 loose stools/ 24 hours) by Day 7 of therapy, in which placebo 
is expected to have a clearance rate of approximately 15-20%.  Sample size calculations for both 
primary endpoints are based on a two-sample, two-sided binomial test for proportions, with alpha 
set to 0.05.  Calculations are performed using the R statistical package (Version 3.0.2; 
http://www.r-project.org).  The primary amebiasis endpoint in this aim is to compare the 
resolution of diarrhea between the two study arms to demonstrate that auranofin increases 


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55 
resolution of diarrhea. For this power analysis, we assume the following: a predicted diarrhea 
resolution rate of at least 80% (based on treatment results with current standard therapy with 
metronidazole, Pherson, 1984) in the auranofin treatment group.   
With these assumptions, assuming 10% attrition (determined from the average of multiple other 
clinical trials in the same districts), a sample size of 68 with amebiasis (34 per arm; 60 completers 
in total) achieves greater than 95% power to detect a difference between the group proportions of 
60%. The high statistical power for the primary endpoint is planned to also ensure adequate power 
for the secondary endpoints.  
For the primary giardiasis endpoint of proportion of subjects with resolution of diarrhea by Day 
5, a similar sample size of 34 per arm will achieve 85% power to detect a 40% difference between 
treatment arms with a predicted clearance rate of 70% (based on response rates to metronidazole 
or tinidazole, Gardiner, 2001) and a 30% clearance rates in placebo. 
11.3 Planned Interim Analyses 
There will be no planned interim analyses for efficacy or futility conducted for this study, but the 
DSMB may modify this during ongoing safety monitoring. The study team will review all adverse 
events by cumulative reports on a monthly basis.  Adverse events will be graded using the 
protocol defined grading system. The DSMB, established by the study team, will monitor this 
study. 
 
In the absence of any SAEs, the DSMB may review the safety data when the study reaches 50% 
enrollment (i.e. 34 subjects enrolled per parasite group). This review will include safety data after 
the 68th subject completes Visit 08.  
11.3.1 Safety Review 
Interim safety review may include enrollment and demographic information, medical history, 
concomitant medications, physical assessments, clinical laboratory values, dosing, and solicited 
and unsolicited AE/SAEs.  Additional data may be requested by the DSMB, and interim statistical 
reports may be generated as deemed necessary and appropriate by DMID.  The DSMB may 
receive data in aggregate and presented by group. The DSMB will meet and review this data at 
scheduled time points or ad hoc as needed during the study as defined in the DSMB charter.  As 
an outcome of each review/meeting, the DSMB will make a recommendation as to the advisability 
of proceeding with study vaccinations (as applicable), and to continue, modify, or terminate the 
study. 


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Additionally, the study will be monitored to determine if any of the halting rules described in 
Section 9.5 are met. 
11.4 Final Analysis Plan 
Analysis Plan for Primary Outcomes. The primary amebiasis endpoint is resolution of diarrhea 
by Day 7. Parasite clearance rates between the placebo and auranofin arms for Entamoeba will 
be compared using a Fisher’s exact test for proportions. Differences in the rates between the two 
groups, along with the odds ratio (OR) and their 95% confidence intervals will be reported.  As a 
secondary analysis, multivariable logistic regression analysis will be performed to study the 
association between clearance rates and intervention arm, adjusting for baseline demographic, 
stratification variables, and clinical characteristics. Variables significantly associated with both 
treatment group and outcome (p<0.10) will be included in a multivariable logistic regression 
model as covariates.   There will be no planned interim analyses for efficacy or futility conducted 
for this study, but the DSMB may modify this during ongoing safety monitoring.  
 
Similarly, for the giardiasis primary endpoint, methods analogous to the amebiasis primary 
endpoint will be used to determine the resolution of diarrhea by Day 5. 
 
Analysis Plan for Secondary Outcomes. The secondary outcome measures are: Entamoeba: 
Proportion of subjects with positive EIA for E. histolytica and positive smear for trophozoites on 
enrollment  with parasitological response (no detection of trophozoites on microscopic exam) by 
Day 7; proportion of subjects with positive EIA for E. histolytica and positive smear for 
trophozoites on enrollment with parasitological response (no detection of trophozoites on 
microscopic exam or negative antigen detection) by Day  3 and 5; rate of decrease of 
trophozoite/cyst load by qPCR in stools by Days 3, 5 and 7; proportion of subjects with negative 
stool antigen test by Days 3, 5, 7, and 14;  proportion of subjects with sustained cure (no detection 
of trophozoites by microscopic exam) at 14 and 28 days; and the proportion of subjects with 
relapse (same strain) or re-infection (new strain) with positive stools at Days 14 and 28 by 
genotyping the initial vs. subsequent strain, and time to resolution of diarrhea.  
 
Giardia: Proportion of subjects with positive EIA for Giardia and positive smear for trophozoites 
on enrollment with parasitological response (no detection of trophozoites on microscopic exam) 
by Day 3 and 5; rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3 and 5; 
proportion of subjects with negative stool antigens by days 3 and ; proportion of subjects with 
sustained cure (no detection of trophozoites by microscopic exam) at 14 and 28 days; the 
proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools 
at Days 14 and 28 by genotyping the initial vs. subsequent strain and time to resolution of diarrhea.  
 


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Methods analogous to the analysis of the primary outcomes, such as Fisher’s exact test, will be 
applied for categorical secondary outcomes (proportion of trophozoite/cyst load in stools from 
enrollment by quantitative PCR at days 3, 5, and 7; and percentage of subjects who have relapsed 
at 14 and at 28 days).  For continuous outcomes, such as count data, t-tests or equivalent non-
parametric alternatives will be considered.  No adjustments will be made for multiple testing of 
secondary outcomes.  Longitudinal modeling, via mixed effects models, will be considered to 
compare the rate of change in loads between the two study arms.  Appropriate treatment by time 
interaction terms will be included. Kaplan-Meier curves and log rank tests will be used to compare 
time to diarrhea resolution between the treatment and control arm for each disease group.   
 
In general, analyses will incorporate the modified intent-to-treat (mITT) principle. Participants 
who are “randomized and receive at least one dose of study medication” will be included in the 
mITT population. Subjects who terminate their treatment regimen and receive metronidazole due 
to worsening symptoms will be imputed as treatment failures for the primary outcome.  That is, 
they will be imputed as still symptomatic by Day 7 in the amebiasis group, and by Day 5 in the 
giardiasis group.  Secondary outcomes measured at Days 3, 5, and 7 will be imputed analogously, 
with the exception of the quantitative trophozoite/cyst load. Multiple imputation may be used to 
adjust for missing data in the primary and secondary analyses on the mITT population.  All results 
will be reported as point estimates (odds ratios or mean differences across groups, as appropriate) 
and interval estimates (95% confidence intervals).  All tests of significance for the secondary 
outcomes will be 2-sided and no adjustments will be made for multiple comparisons.  A p-value 
of 0.05 will be considered statistically significant.  The Statistical Analysis Plan (SAP) will 
provide further details. In case the language in this section differs from the language in the SAP, 
the SAP takes precedence. 
In the event that recruitment and follow-up for either the Giardiasis arm or the Amebiasis arm is 
completed ahead of the other, the primary and secondary endpoints may be analyzed for that arm 
using the specified methods.  This report will be considered the interim CSR and will contain 
detailed unblinded information about the participants, their immunological response to treatment 
as well as their side effects and any laboratory abnormalities.  The interim CSR will be followed 
with a release of data corresponding to the report.  In order to maintain the blind and allow 
appropriate follow up of the remaining arm, no endpoint data for the remaining arm will be 
included in the interim CSR and no data related to that arm will be released.  An amendment to 
the interim CSR will provide primary and secondary analysis of the remaining arm when all 
primary and secondary endpoint data are available for the second arm.  The official reports will 
be drafted by Emmes and reviewed by the PI and DMID prior to finalization.  Additional 
information regarding the methods will be provided in the study SAP.     
The purpose of this study is to assess the efficacy of auranofin (Ridaura) for the treatment of 
amebiasis and giardiasis and the timely release of this information, once available, will support 
improved clinical care.  It is anticipated that the results of this study will be presented to the 


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scientific community via oral presentations at meetings and written publications in scientific 
journals following finalization of the interim CSR. The data to be presented and the authorship 
will be discussed between partners before any official communication. 


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12 
SOURCE DOCUMENTS AND ACCESS TO SOURCE 
DATA/DOCUMENTS 
The site will maintain appropriate medical and research records for this trial, in compliance with 
ICH E6, Section 4.9, and regulatory and institutional requirements for the protection of 
confidentiality of subjects.  The site will permit authorized representatives of the DMID, its 
designees, and appropriate regulatory agencies to examine (and when required by applicable law, 
to copy) clinical study records for the purposes of quality assurance reviews, audits, monitoring 
and evaluation of the study safety and progress.  These representatives will be permitted access 
to all source data, which include, but are not limited to, hospital records, clinical and office charts, 
laboratory notes, memoranda, evaluation checklists, pharmacy dispensing records, recorded data 
from automated instruments, copies or transcriptions certified after verification as being accurate 
and complete, microfiches, photographic negatives, microfilm or magnetic media, x-rays, and 
subject files and records kept at the pharmacy, at the laboratories, and medico-technical 
departments involved in the clinical trial.  Source data are all information, original records of 
clinical findings, observations, or other activities in a clinical trial necessary for the reconstruction 
and evaluation of the trial.  Data collection forms will be derived from the eCRFs and be provided 
by the SDCC. 
 
 


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13 
QUALITY CONTROL AND QUALITY ASSURANCE 
Following a written DMID-accepted site clinical quality management plan (CQMP), the 
investigational site is responsible for conducting routine quality assurance and quality control 
activities to internally monitor study progress and protocol compliance.  
 
The PI will provide direct access to all trial-related sites, source data/documents, and reports for 
the purpose of monitoring and auditing by the sponsor, and inspection by local and regulatory 
authorities. The PI will ensure all study personnel are appropriately trained and applicable 
documentation is maintained on site. Staff Training will include Institution-specific Training, 
Protocol-specific Training, and DMID-specific training.  
 
DMID-designated clinical monitors will verify that the clinical trial is conducted and data are 
generated, documented (recorded), and reported in compliance with the protocol, ICH/GCP 
guidelines, and the applicable regulatory requirements. Clinical monitoring reports will be 
submitted to DMID. 
The Statistical and Data Coordinating Center will implement quality control procedures 
beginning with the data entry system and generate data quality control checks that will be run on 
the database. Any missing data or data anomalies will be communicated to the site(s) for 
clarification and resolution. 
 


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14 ETHICS/PROTECTION OF HUMAN SUBJECTS 
14.1 Ethical Standard 
The principal investigator will ensure that this trial is conducted in full conformity with principles 
of the Belmont Report:  Ethical Principles and Guidelines for the Protection of Human Subjects 
of Research of the National Commission for the Protection of Human Subjects of Biomedical and 
Behavioral Research (April 18, 1979) and codified in 45 CFR 46, 21 CFR 50 and 56, and ICH 
E6; 62 Federal Regulations 25691 (1997), if applicable.  The site principal investigator’s 
Institution will hold a current Federal Wide Assurance (FWA) issued by the Office of Human 
Research Protection (OHRP) for federally funded research. 
14.2 Institutional Review Board/Ethics Committee 
UCSD is the lead institution in this project and where the PI is located. Quantitative PCR testing 
of positive stool specimens will be performed at UCSD. Although no direct human subjects 
research will occur at this institution, UCSD will be responsible for all regulatory matters in 
conjunction with our collaborators. This includes coordination of all IRB approvals, quality 
assurance, data safety and monitoring and protocol development. The International Centre for 
Diarrhoeal Research, Bangladesh (icddr,b) and  Rajshahi Medical College Hospital and Clinic  
are the  sites where human subjects research procedures will be performed, with the option for 
home visits also conducted as indicated. This includes screening, recruiting, and collecting all 
data/samples and following up subjects as detailed in Section 7.  The icddr,b Ethics Committee 
will also review and approve study protocols which is also is used for approval of joint Rajshahi 
Medical College protocol.  Dr. Petri’s lab at the University of Virginia will perform off site 
molecular testing of specific strains of Giardia or E. histolytica to identify relapse or re-infection.  
14.3 Informed Consent Process 
Informed consent is a process that is initiated prior to the individual’s agreeing to participate in 
the study and continuing throughout the individual’s study participation.  Extensive discussion of 
risks and possible benefits of this therapy will be provided to the subjects and their families.  
Consent forms describing in detail the Study Agent, auranofin, and risks are given to the subject 
and written documentation of informed consent is required prior to starting study 
agent/intervention.  Consent forms will be IRB approved and the subject will be asked to read 
and review the document. Upon reviewing the document, the investigator will explain the 
research study to the subject and answer any questions that may arise.  The subjects will sign the 
informed consent document prior to any procedures being done specifically for the study.  The 
subjects should have sufficient opportunity to discuss the study and process the information in 


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the consent process prior to agreeing to participate.  The subjects may withdraw consent at any 
time throughout the course of the trial.  A copy of the informed consent document will be given 
to the subjects for their records.  The rights and welfare of the subjects will be protected by 
emphasizing to them that the quality of their medical care will not be adversely affected if they 
decline to participate in this study. 
The informed consent procedure is designed to maximize the potential subject’s comprehension 
of study procedures and to ensure that participation is voluntary.  Before a subject is enrolled in 
the study, the purpose, the procedures to be followed, and the risks and benefits of participation 
will be explained by the clinic staff, and signed informed consent from the study subject will be 
obtained. For those subjects unable to read or sign the consent documents, a literate witness will 
be present to sign on behalf of the subject after the subject has given oral consent. In addition to 
the signature of a literate witness, agreement of subjects who are illiterate will be indicated by 
including his/her thumbprint on the consent form, in accordance with the WHO guidelines on the 
consent 
process 
for 
illiterate 
individuals 
(http://www.who.int/rpc/research_ethics/Process_seeking_IF_ printing.pdf). 
Clinic staff will be trained to respond to the subject’s questions and concerns. Details of study 
participation, including the need for follow-up contact, will be described in both the consent form 
and explained verbally. The research staff who will be responsible for obtaining informed consent 
will assess whether the potential subject has understood critical aspects of the study and consent 
form by asking key questions (e.g., “How much time will this take you?”; “What are the possible 
risks to you?”). Errors will be corrected and these potential subjects will then be asked if they 
need further clarification. If, after further attempts to clarify any misunderstandings, we determine 
that they may not fully comprehend the critical aspects of the study, they will not be enrolled. If 
a potential subject decides he/she does not wish to participate, his/her decision will be honored 
regardless of how well they comprehend the study information. The subject may refuse to 
participate or withdraw at any time without jeopardy to the medical care they regularly receive. 
The investigator will explain to the subject that only parasite DNA samples will be collected and 
analyzed and that no human genetic material will be collected.  
A copy of the consent form, which includes a description of the study, will be provided to all 
subjects, which includes contact information of the main investigators in Dhaka and Rajshahi 
Bangladesh, and the U.S. Consent forms will be written in simple language at a fourth-grade 
reading level, and will be translated into Bengali. In the case of subjects with low or no literacy, 
the methods described above of: 1) Verbally reviewing the consent form; and 2) Giving a simple 
test about points made in each paragraph of the consent, should open enrollment to all regardless 
of literacy level.  Participants will be encouraged to ask questions and retested if they do not fully 
understand any of the study protocols. Prior to study implementation we will receive approval 
from the UCSD Institutional Review Board (IRB) Committee on Human Research, the icddr,b 


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Ethical Review Committee in Bangladesh and the UVa Institutional Review Board (IRB) for 
Health Sciences Research. 
Participation in this study is completely voluntary, and the alternative to study participation is not 
to participate. Participants may refuse to participate or withdraw at any time without jeopardy to 
the medical care they regularly receive. Treatment for diarrhea is available at health clinics in 
Bangladesh. If a subject chooses not to participate in this study or is deemed ineligible to 
participate, he/she may ask the study staff for a referral to their nearest clinic. Whether they join 
this study or not, subjects may also ask the study staff for information about infectious diseases, 
such as Giardia and E. histolytica. 
14.4 Exclusion of Women, Minorities, and Children  
(Special Populations) 
This study will be inclusive of all adults who meet the inclusion/exclusion criteria, regardless of 
religion, sex, or ethnic background. Women who are pregnant or breastfeeding, women of 
childbearing potential who do not agree to practice effective contraception, and children are 
excluded for safety reasons. Should the outcome of this study be deemed acceptable, additional 
trials may be initiated in other populations. 
14.5 Subject Confidentiality 
Subject confidentiality is strictly held in trust by the participating investigators, their staff, and 
the sponsor(s) and their agents.  This confidentiality is extended to cover testing of biological 
samples and genetic tests in addition to the clinical information relating to participating subjects. 
The study protocol, documentation, data, and all other information generated will be held in strict 
confidence.  No information concerning the study or the data will be released to any unauthorized 
third party without prior written approval of the sponsor. 
The study monitor or other authorized representatives of the sponsor may inspect all documents 
and records required to be maintained by the investigator, including but not limited to, medical 
records (office, clinic, or hospital) and pharmacy records for the subjects in this study.  The 
clinical study site will permit access to such records. 
To guard confidentiality, only the subject's identification number will appear on screening forms, 
questionnaires, databases and biological samples. The key to subjects’ identification numbers will 
be encrypted in a computer file, which will be locked in the field coordinator’s office. Only the 
site PI and field coordinator will be able to un-encrypt the computer file. Contact information for 


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the subject so that s/he can be located at follow-up visits will be stored in locked file cabinets in 
locked offices at the clinic, and will be destroyed once follow-up is completed. 
14.6 Study Discontinuation 
If the trial is discontinued, subjects who had previously signed the informed consent form, and 
were randomized and treated will continue to be followed for safety assessments.  No further 
study treatments will be administered. 
14.7 Future Use of Stored Specimens 
At the time of informed consent, the Investigator will seek permission from subjects for use of 
their remaining sample for possible future research studies, investigating parasitic disease, such 
as the effect of parasitic infections and/or auranofin on the stool microbiome. All samples will be 
stored in a freezer at the icddr,b or Rajshahi Medical College Hospital for 3-4 days maximum, 
except for those that test positive for Giardia or Entamoeba. Samples from Rajshahi Medical 
College Hospital will be sent to icddr,b first , then to UCSD/UVA. These will be stored in the 
icddr, b freezer for the entire duration of the study, after which they will be stored indefinitely at 
the Fisher Repository Samples and may be shared with other investigators at other institutions.  
The samples will not be sold or used directly for production of any commercial product.  No 
human genetic material will be collected or stored and no human genetic tests will be performed 
on any samples.  Each sample will be de-identified and encoded (labeled) only with a barcode 
and a unique tracking number to protect subject’s confidentiality. 
There are no benefits to subjects in the collection, storage and subsequent research use of 
specimens.  Reports about future research done with subject’s samples will NOT be kept in their 
health records. 
Subjects may be given the option to decide if they want their samples to be used for future research 
or have their samples destroyed at the end of the trial.  The subject’s decision can be changed at 
any time prior to the end of the trial by notifying the study doctors or nurses in writing.  However, 
if the subject originally consents to future use and subsequently changes his/her decision, any data 
from a previously collected sample may still be used for this research. 


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15 DATA HANDLING AND RECORD KEEPING 
The investigator is responsible to ensure the accuracy, completeness, legibility, and timeliness of 
the data reported.  All source documents should be completed in a neat, legible manner to ensure 
accurate interpretation of data.  Black ink is required to ensure clarity of reproduced copies.  When 
making changes or corrections, cross out the original entry with a single line, and initial and date 
the change.  DO NOT ERASE, OVERWRITE, OR USE CORRECTION FLUID OR TAPE ON 
THE ORIGINAL. 
Copies of the electronic CRF (eCRF) will be provided for use as source documents and 
maintained for recording data for each subject enrolled in the study.  Data reported in the eCRF 
derived from source documents should be consistent with the source documents or the 
discrepancies should be explained 
DMID and/or it designee will provide guidance to investigators on making corrections to the 
source documents and eCRF. 
15.1 Data Management Responsibilities 
All data collection forms and laboratory reports must be reviewed by the clinical team and data 
entry personnel, who will ensure that they are accurate and complete.  Adverse events must be 
recorded on the appropriate data collection form, assessed for severity and relationship, and 
reviewed by the site principal investigator or appropriate sub-investigator. 
Data collection is the responsibility of the study personnel at the participating sites under the 
supervision of the respective site principal investigator.  During the study, the site principal 
investigator must maintain complete and accurate documentation for the study. 
EMMES, The SDCC for this study will be responsible for data management, quality review, 
analysis, and reporting of the study data. 
15.2 Data Capture Methods 
Clinical (including, but not limited to, AE/SAEs, concomitant medications, medical history, 
physical assessments, and clinical laboratory values), data will be entered into a 21 CFR 11-
compliant Internet Data Entry System provided by the SDCC.  The data system includes password 
protection and internal quality checks, such as automatic range checks, to identify data that appear 
inconsistent, incomplete, or inaccurate.  Clinical data will be entered directly from the data 
collection forms completed by the study personnel. 


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15.3 Types of Data 
Data for this study will include safety, laboratory, and outcome measures. 
15.4 Timing/Reports 
The study team will review all adverse events by cumulative reports of the treatment arms on a 
monthly basis. 
 
Asymptomatic subjects (34) enrolled under Version 1-4 of the protocol will have their data 
reviewed and clinical database locked for analysis first and a separate report generated.  
A final report will then be prepared following the availability of all the safety and outcome data 
for one study arm with symptomatic patients (Giardiasis or Amebiasis). An amendment to the 
final report will be produced following the availability of all safety and outcome data for the 
remaining study arm.  Interim statistical reports may be generated as deemed necessary and 
appropriate by DMID.  Safety and immunogenicity summary reports may be generated for the 
DSMB. 
After full analysis and final reporting is complete, and upon request and DMID approval, the 
SDCC will provide the site with a summary of results by treatment group and/or subject treatment 
assignments.  In this regard, the participating site requesting such information to share with study 
subjects must do so in compliance with their respective IRB guidelines. 
15.5 Study Records Retention 
Study records and reports, including, but not limited to, case report forms (CRFs), source 
documents, informed consent forms (except for future use informed consent forms), laboratory 
test results, and medication inventory records, shall be retained for 2 years after a marketing 
application is approved for the drug; or, if an application is not approved for the drug, until 2 
years after shipment and delivery of the drug for investigational use is discontinued and FDA has 
been so notified.  The site must contact DMID for authorization prior to the destruction of any 
study records.  Informed consent forms for future use will be maintained as long as the sample 
exists. 
15.6 Protocol Deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, Good Clinical Practice 
(GCP), or Manual of Procedures requirements.  The noncompliance may be either on the part of 
the subject, the investigator, or the study site staff.  As a result of deviations, corrective actions 
are to be developed by the site and implemented promptly. The site will use continuous vigilance 


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to identify and report deviations according to the guidelines of the IND sponsor, if applicable. 
Protocol deviations will be sent to the local IRB/IEC per their guidelines.   
 
These practices are consistent with ICH E6: 
4.5 Compliance with Protocol, Sections 4.5.1, 4.5.2, and 4.5.3 
5.1 Quality Assurance and Quality Control, Section 5.1.1 
5.20 Noncompliance, Sections 5.20.1, and 5.20.2 
It is the responsibility of the site principal investigator and other study personnel to use continuous 
vigilance to identify and report deviations within five working days of identification of the 
protocol deviation, or within five working days of the scheduled protocol-required activity.  All 
deviations must be promptly reported to DMID, via the SDCC’s Emmes Advantage eClinical  
system.  
All protocol deviations, as defined above, must be addressed in study subject data collection 
forms.  A completed copy of the DMID Protocol Deviation Form must be maintained in the 
Regulatory File, as well as in the subject's chart.  Protocol deviations must be sent to the local 
IRB/IEC per its guidelines.  The site principal investigator and other study personnel are 
responsible for knowing and adhering to their IRB requirements. 
 


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16 PUBLICATION POLICY 
All investigators funded by the NIH must submit or have submitted for them to the National 
Library of Medicine’s PubMed Central (http://www.ncbi.nlm.nih.gov/pmc/) an electronic version 
of their final, peer-reviewed manuscripts upon acceptance for publication, to be made publicly 
available no later than 12 months after the official date of publication.  The NIH Public Access 
Policy ensures the public has access to the published results of NIH funded research.  It requires 
investigators to submit final peer-reviewed journal manuscripts that arise from NIH funds to the 
digital archive PubMed Central upon acceptance for publication.  Further, the policy stipulates 
that these papers must be accessible to the public on PubMed Central no later than 12 months 
after publication. 
Refer to: 
• 
NIH Public Access Policy, http://publicaccess.nih.gov/ 
• 
NIH OER Grants and Funding, http://grants.nih.gov/grants/oer.htm 
Following completion of the study, the lead principal investigator is expected to publish the 
results of this research in a scientific journal.  The International Committee of Medical Journal 
Editors (ICMJE) member journals have adopted a trials-registration policy as a condition for 
publication.  This policy requires that all clinical trials be registered in a public trials registry such 
as ClinicalTrials.gov* (http://clinicaltrials.gov/), which is sponsored by the National Library of 
Medicine.  Other biomedical journals are considering adopting similar policies. 
The ICMJE defines a clinical trial as any research project that prospectively assigns human 
subjects to intervention or comparison groups to study the cause-and-effect relationship between 
a medical intervention and a health outcome.  Studies designed for other purposes, such as to 
study pharmacokinetics or major toxicity (e.g., Phase I trials), would be exempt from this policy. 
In the event that either the Giardia or E. histolytica portion of the trial is completed earlier than 
the other cohort, data from the completed cohort may be released after the database from that 
portion of the trial is locked. 
It is the responsibility of DMID to register this trial in an acceptable registry.  Any clinical trial 
starting enrollment after 01 July 2005 must be registered on or before subject enrollment.  For 
trials that began enrollment prior to this date, the ICMJE member journals will require registration 
by 13 September 2005, before considering the results of the trial for publication. 
For trials in which DMID is not the IND/IDE sponsor, or there is no IND/IDE, and DMID does 
not provide data management services, it is the responsibility of the investigator to register the 


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trial and post results in compliance with Public Law 110-85, the Food and Drug Administration 
Amendments Act of 2007 (FDAAA). 


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of diversity and a genetic component to Entamoeba histolytica virulence. BMC Microbiol. 
12:151-65.  
Gilman RH, Brown KH, Visvesvara GS, Mondal G, Greenberg, B, Sack RB, Brandt F, Khan, 
MU. 1985. Epidemiology and serology of Giardia lamblia in a developing country. Roy Soc Trop 
Med Hyg. 79:469-73.  
Gut J, Ang KKH, Legac J, Arkin MR, Rosenthal PJ, McKerrow JH. 2011. An image-based assay 
for high throughput screening of Giardia lamblia. J. Microbiol. Methods. 84:398-405.  
Haque, R., Ali, I.M. Sack, R.B., Farr, B.M., Ramakrishnan, G., Petri, W.A. 2001.Amebiasis and 
mucosal IgA antibody against the Entamoeba histolytica adherence lectin in Bangladeshi 
children. J. Infect. Dis. 183:1787-93. 
Haque, R., Duggal, P., Ali, I.M., Hossain, M.B., Mondal, D., Sack, R.B., Farr, B.M., Beaty, T.H., 
and Petri, W.A. 2002. Innate and acquired resistance to amebiasis in Bangladeshi children. J. 
Infect. Dis. 186:547-52. 
Haque, R., Huston, C.D., Hughes, M., Houpt, E. and Petri, W.A. 2003. Amebiasis. N. Engl. J. 
Med. 348:1565-73.  
Haque, R., Mondal, D., Duggal, P., Kabir, M., Roy, S., Farr, B.M., Sack, R.B., and Petri, W.A.  
2006. Entamoeba histolytica infection in children and protection from subsequent amebiasis. 
Infect. Immun.74:904-909. 
Haque R, Roy S, Siddique A, Mondal U., Rahman SM, Mondal D, Houpt E, and Petri WA. 2007. 
Multiplex real-time PCR assays for detection of Entamoeba histolytica, Giardia intestinalis, and 
Cryptosporidium spp. Am. J.Trop. Med. Hyg. 76:713-7, 2007. 


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14 APR 2020 
 
72 
Haque R,  Mondal D,  Karim MA, Molla IH,  Rahim MA, Faruque ASG, Ahmad N,  
Kirkpatrick BD,  Houpt E,  Snider C, and  Petri  WA Jr. 2009. Association of Common Enteric 
Protozoan Parasites with Severe Diarrhea in Bangladesh:  A Prospective Case-Control Study.  
Clin Infect Dis, 48: 1191- 97.  
Ilari, A. Baiocco P, Messori L, Fiorillo A, Boffi A, Gramiccia M, Muccio TD, Coilotti G. 2012.A 
gold-containing drug against parasitic polyamine metabolism: the X-ray structure of 
trypanothione reductase from Leishmania infantum in complex with auranofin reveals a dual 
mechanism of enzyme inhibition. Amino Acids42, 803–11. 
Kuntz, A.N.   Davioud-Charvet E, Sayed AA, Califf LL, Dessolin J, Amer ESJ, Williams, D. 
2007. Thioredoxin glutathione reductase from Schistosoma mansoni: an essential parasite enzyme 
and a key drug target. PLoS Med.4, e206.  
Lemee, V, Zharia, I, Nevez, G., Rabodonirina, M. Brasseur, P., Ballet, J, Favennec L. 2000. 
Metronidazole and albendazole susceptibility of 11 clinical isolates of Giardia duodenalis from 
Frane. H. Antimicrob. Chemo. 46:819-21.  
Lobanov AV, Gromer S, Salinas G, Gladyshev VN. 2006. Selenium metabolism in Trypanosoma; 
characterization of selenoproteomes and identification of a Kinetoplastida-specific selenoprotein. 
Nucl. Acids. Res. 34: 4012-24. 
McAuley JB, Herwaldt BL, Stokes SL, Becher JA, Roberts JM, Michelson MK, Juranek DD. 
1002. Diloxanide furoate for treating asymptomatic cyst passers: 14 years’ experience in the 
United States. Clin.Infect.Dis 15:464-8. 
Pherson, P.O. and Bengtsson, E. 1984. A long-term follow up study of amebiasis treated with 
metronidazole. Scand. J. Infect. Dis. 16:195-8. 
Rossignol, JF, Ayoub A, Ayers MS. 2001. Treatment of diarrhea caused by Giardia intestinalis 
and Entamoeba histolytica or E. dispar: a randomized, double blind, placebo-controlled study of 
Nitazoxanide. J. Infect. Dis. 184:381-4. 
Roy S, Kabir M, Mondal D, Ali IK, Petri WA, Haque R. Real-time PCR assay for diagnosis of 
Entamoeba histolytica infection. J ClinMicrobiol. 43:2168-72.  
Savioli, Smith H, and Thompson A 2006.Giardia and Cryptosporidium join the ‘Neglected 
Diseases Initiative’. Trends in Parasitol. 22:203-8.  
Solaymani-Mohammed, S. Genkinger, J.M., Loffredo, CA, Singer, S.M. 2010. Meta-analysis of 
the effectives of albendazole compared with metronidazole as treatment of infections with 
Giardia duodenalis. PLoS Neg. Trop. Dis. 4(5): e682.  


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73 
Taniuchi, M., Verweij, J.J., Noor, Z., Sobuz, S.U., van Lieshout, L., Petri, W.A., Haque, R., and 
Houpt. E.R. 2011. High throughput multiplex PCR and probe-based detection with Luminex 
beads for seven intestinal parasites. 2011. Am. J. Trop. Med. Hyg. 84:332-7.  
Tejman-Yarden N, Miyamoto Y, Leitsch D, Santini J, Denath A, Gut J, McKerrow J, Reed SL, 
Eckman L. A reprofiled drug, auranofin, is effective against metronidazole-resistant Giardia 
lamblia, Antimicrob Agent Chemothr. 57(5):2029-35., 2013.  
Upcroft P, Upcroft JA. 2001. Drug targets and mechanisms of resistance in the anaerobic 
protozoa. ClinMicrobiol. Rev. 14:150-64. 
Wassmann, C., Hellberg, A., Tannich, E., and Bruchhaus, I. 1999. Metronidazole resistance in 
the protozoan parasite Entamoeba histolytica is associated with increased expression of iron-
containing superoxide dismutase and peroxiredoxin and decreased expression of ferredoxin 1 
and flavin reductase. J. Biol. Chem. 274:26051-26056. 
Wright JM, Dunn LA, Upcroft P, Upcroft JA. 2003. Efficacy of antigiardial drugs. Expert Opin 
Drug Saf. 2:529-41.  
Yoder, JS, Harral, C., Beach, MJ. 2010. Giardiasis surveillance- United States, 2006-8, MMWR. 
59:15-25. 
 


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A-1 
APPENDIX A:   SCHEDULE OF EVENTS 
Procedure 
Visit* 
Number 
00A 
00B 
01 
02 
03 
04 
05 
062 
07 
08 
09 
102 
Study Day  
-4 
-4 
1 
2 
3 
4 
5 
6 
7  
14 
28 
30 
Window 
(days) 
-4 to1 
-4 to 
-1 
+16--- 
--- 
--+1- 
--- 
+16--- 
--- 
+1 
+/-3 
+/-3 
 
+/-3 
Review 
Eligibility 
 
X 
 
 
X 
 
 
 
 
 
 
 
 
 
Informed 
Consent 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Demographic 
Data 
 
X 
 
 
 
 
 
 
 
 
 
 
 
Medical History 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
Targeted 
Physical Exam3 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
Vital signs4 
 
 
X 
X 
 
 
 
 
 
 
 
 
 
Concomitant 
Medications 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
Randomization/
Enrollment 
 
 
 
X 
 
 
 
 
 
 
 
 
 
Assessment for 
side effects of 
treatment 
 
 
 
 
 
 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
X 
 
AEs/SAEs 
 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
Urine for 
pregnancy 
testing 
 
 
X 
 
 
 
 
 
 
 
 
 
 
Treatment 
(giardiasis 
subjects) 
 
 
 
X 
X 
X 
X 
X 
 
 
 
 
 


DMID Protocol 15-0015                                                            Version 8.0 
14 APR 2020 
 
 
A-2 
Treatment (E. 
histolytica)  
 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
 
 
 
Procedure 
 
 
Visit 
Number 
00A 
00B 
01 
02 
03 
04 
05 
063 
07 
08 
09 
104 
Study Day  
-4 
-3 
1 
2 
3 
4 
5 
6 
7  
14 
28 
30 
Window 
(days) 
-4 to 
1 
-3 to 
-1 
+18--- 
--- 
--+18- 
--- 
+18--- 
--- 
+1 
+/-3 
+/-3 
 
+/-3 
Urine Dipstick 
for Proteinuria 
 
 
 
X 
 
 
 
 
 
 
 
X 
 
 
 
Complete 
Blood Count5 
 
 
 
X 
 
 
 
 
 
 
X 
 
 
 
BUN 
 
 
X 
 
 
 
 
 
 
X 
 
 
 
Creatinine 
 
X 
 
 
 
 
 
 
X 
 
 
 
ALT 
 
X 
 
 
 
 
 
 
X 
 
 
 
AST 
 
X 
 
 
 
 
 
 
X 
 
 
 
Stool 
specimen6 
 
X 
 
X 
 
X 
 
X 
 
X 
X 
X 
 
Blood volume 
 
 
17 
 
 
 
 
 
 
17 
 
 
 
Blood volume 
totals 
 
 
17 
 
 
 
 
 
 
34 
 
 
 
Standard of 
Care Treatment 
 
 
 
 
 
 
 
 
 
 
 
 
X 
 
*Subjects will be enrolled at the icddr,b or Rajshahi Hospitals and have follow-up visits until their diarrhea has resolved. Subsequent follow-up visits may be 
held at the icddr,b or Rajshahi Hospital, or local (to Rajshahi) clinic or their home except for Visit 07 which must be performed at icddr,b or Rajshahi Hospital or 
Clinic because blood drawing is required. 
1Visit 06 will only be conducted for subjects receiving treatment for amebiasis. 
2Visit 10 will only be conducted for subjects who remain positive for amebiasis or giardiasis at Day 28 and have not previously received standard of care 
treatment.  
3Targeted physical exam may also include observations for rashes, hair loss, urticaria or glossitis, and an abdominal exam. 
4Vital signs (oral temperature, pulse, blood pressure) 
5Complete blood count includes white blood cells, hemoglobin, and platelet count.  
6Window of + 1 day for stool specimens after (and including) Day 1. Date of collection will be noted. The window does not apply to treatment administration or 
other required assessments. 


DMID Protocol 15-0015                                                            Version 8.0 
14 APR 2020 
 
 
A-3 
7Amebiasis patients only 


DMID Protocol 15-0015                                            Version 8.0 
14 APR 2020 
 
A-4 
 
APPENDIX B:LAB PROCESSING FLOW SHEET: CLINICAL/ 
SAFETY LAB TESTS 
Protocol # 15-0015 
 
Instructions- All specimens must be recorded on a form similar to the one below including the visit time/study ID number, tube type, 
specimen test, processing and storage instructions 
Visit/Time of 
Draw 
Tube Type/Quantity 
Test/Assay/Derivative 
Processing/Aliquot/Storage/Shipping 
Screening 
 
Day -4 to -
1, 
Visit 00A 
 
Stool collection vial for Stool 
Evaluation 
 
Rapid Stool EIA for Giardia 
or E. histolytica 
EIA stool antigen test if 
positive  
Wet mount 
Concentration and 
Trichrome staining 
Qualitative PCR for Giardia 
and Entamoeba 
Stools kept cold and shipped daily to icddr,b or Rajshahi 
hospital- See Lab SOPs 
 
 
Screening  
 
Day -4 to -1, 
Visit 00B 
Urine collection vial 
Dip stick for protein 
Pregnancy testing 
Performed in icddr,b or Rajshahi Hospital or designated 
lab,  
Purple top tube (7mls) 
Complete blood count 
Kept at room temp and transferred same day to lab for 
analysis 
Serum separator tube (10 ml) 
BUN, creatinine, SGOT, 
SGPT 
Kept at room temp and transferred same day to lab for 
analysis 
Left over serum aliquoted and kept at -80oC 
Enrollment 
Day 1,  
Visit 01 
Stool collection vial for Full 
Stool Evaluation (+ 1 Day) 
 
 
 
 
 
 
 
 
 
 
 
 
 
Antigen testing for Giardia 
or E. histolytica 
 
Wet mount 
Concentration and 
Trichrome staining 
 
Qualitative PCR for Giardia 
and Entamoeba 
 
Quantitative PCR for 
Giardia or Entamoeba in 
positive stools 
 
Specimens processed as above 
 
Processed as above 
 
 
Fixed stool stored at room temp  
 
 
Processed as above 
 
Extracted DNA from stool obtained before first 
auranofin dose. Shipped every 1-3months to UCSD for 
quantitative PCR   
Day 3,  
Visit 03  
Stool collection vial for Full 
Stool Evaluation (+1 day) 
Refer to Day 1 (Not 
performed on patients who 
receive metronidazole prior 
to completing dosing 
regimen) 
Refer to Day 1 
Day 5,  
Visit 05 
Stool collection vial for Full 
Stool Evaluation 
(+ 1 day) 
 
Refer to Day 1 (Not 
performed on patients who 
receive metronidazole prior 
Refer to Day 1 
 


DMID Protocol 15-0015                                            Version 8.0 
14 APR 2020 
 
A-5 
 
to completing dosing 
regimen) 
Day 7, 
Visit 07 
 
Stool collection vial for Full 
Stool Evaluation (+1 Day) 
 
Refer to Day 1 (Not 
performed on patients who 
receive metronidazole prior 
to completing dosing 
regimen) 
Refer to Day 1 
Urine Collection Vial 
Urine Dip stick for protein 
Refer to Screening Visit 00B 
Purple top tube (7mls) 
Complete blood count and 
differential  
Refer to Screening Visit 00B 
SST (10 ml) 
BUN, Cr, SGOT, SGPT,  
Refer to Screening Visit 00B 
Day 14, 
Visit 08 
+/- 3 days 
Stool collection vial for Full 
Stool Evaluation 
Refer to Day 1 
If positive at Day 14, 
perform parasite genotyping. 
(Not performed on patients 
who receive metronidazole 
prior to completing dosing 
regimen) 
Refer to Day 1 
Stored DNA sent to UVA for parasite genotyping every 
1-3 months 
Day 28, 
Visit 09 
+/- 3 days 
Stool collection vial for Full 
Stool Evaluation 
Refer to Day 1 
If positive at Day 14, 
perform parasite genotyping. 
(Not performed on patients 
who receive metronidazole 
prior to completing dosing 
regimen) 
Refer to Day 1 
Stored DNA sent to UVA for parasite genotyping every 
1-3 months 
 
 
 


DMID Protocol 15-0015                                            Version 8.0 
14 APR 2020 
 
A-6 
 
APPENDIX C: ICDDR,B NORMAL LABORATORY VALUES 
(APPLICABLE FOR BOTH SITES) 
 
Male 
Female 
White Blood cells (109/L) 
4.0 –11.0   
same 
Platelets (109/L) 
150.0 –  450.0 
same 
Hemoglobin (gm/dL) 
12.5 –17.5 
11.5 – 16.5  
Blood urea nitrogen (mg/dL) 
5 –24  
same 
Creatinine (umol/L) 
 53-106 
44-97 
AST/SGOT (U/L) 
0.01 – 38.0 
0.01 – 32.0 
ALT/SGPT (U/L) 
0.01 – 41.0 
0.01 – 31.0 
 
 

SAP_001.pdf: 
CLINICAL RESEARCH IN INFECTIOUS DISEASES 
STATISTICAL ANALYSIS PLAN 
for 
DMID Protocol: 15-0015 
Study Title: 
Phase IIa Randomized, Single-blinded, Placebo-controlled Clinical Trial of the 
Reprofiled Drug Auranofin for GI Protozoa 
 
NCT02736968 
 
Version 2.0 
 
 
 
DATE: 29 APRIL 2020 
THIS COMMUNICATION IS PRIVILEGED AND CONFIDENTIAL 
 


Statistical Analysis Plan - DMID Protocol: 15-0015  
Version 2.0 
29 April 2020 
- ii - 
PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
STUDY TITLE: Phase IIa Randomized, Single-Blinded, Placebo-Controlled Clinical Trial of 
the Reprofiled Drug Auranofin for Gi Protozoa 
Protocol Number Code: 
DMID Protocol: 15-0015 
Development Phase: 
Phase IIa 
Products: 
Auranofin 
Form/Route: 
One dose (two capsules containing 3 mg each) per day orally of auranofin 
for seven days to treat amebiasis; one dose (two capsules containing 3 mg 
each) per day of auranofin for five days to treat giardiasis. 
Indication Studied: 
Amebiasis and giardiasis 
Sponsor: 
Division of Microbiology and Infectious Diseases 
National Institute of Allergy and Infectious Diseases 
National Institutes of Health 
Clinical Trial Initiation Date: 
 
Clinical Trial Completion Date: 
 
Date of the Analysis Plan: 
29 April 2020 
Version Number: 
2.0 
This study was performed in compliance with Good Clinical Practice. 
Information contained in this publication is the property of Division of Microbiology and Infectious Diseases and is confidential. This 
information may not be disclosed to third parties without written authorization from Division of Microbiology and Infectious Diseases. This 
report may not be reproduced, stored in a retrieval system or transmitted in any form or by any means - electronic, mechanical, recording or 
otherwise - without the prior authorization from Division of Microbiology and Infectious Diseases. This document must be returned to Division 
of Microbiology and Infectious Diseases upon request. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
Version 2.0 
29 April 2020 
- iii - 
PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
TABLE OF CONTENTS 
STUDY TITLE: .............................................................................................................................. II 
TABLE OF CONTENTS ...............................................................................................................III 
LIST OF ABBREVIATIONS ....................................................................................................... VI 
1. 
PREFACE .....................................................................................................................8 
2. 
INTRODUCTION ........................................................................................................9 
2.1. 
Purpose of the Analyses ..............................................................................................12 
3. 
STUDY OBJECTIVES AND ENDPOINTS ..............................................................13 
3.1. 
Study Objectives .........................................................................................................13 
3.1.1. 
Objectives for the Asymptomatic Population .............................................................13 
3.1.2. 
Objectives for the Symptomatic Population ...............................................................14 
3.2. 
Endpoints ....................................................................................................................15 
3.2.1. 
Asymptomatic Population Endpoints .........................................................................15 
3.2.2. 
Symptomatic Population Endpoints ...........................................................................15 
3.3. 
Study Definitions and Derived Variables ...................................................................16 
4. 
INVESTIGATIONAL PLAN .....................................................................................18 
4.1. 
Overall Study Design and Plan ...................................................................................18 
4.2. 
Discussion of Study Design, Including the Choice of Control Groups ......................19 
4.3. 
Selection of Study Population ....................................................................................19 
4.3.1. 
Asymptomatic Population ..........................................................................................19 
4.3.1.1. 
Inclusion Criteria, Asymptomatic Population ............................................................19 
4.3.1.2. 
Exclusion Criteria, Asymptomatic Population ...........................................................20 
4.3.2. 
Symptomatic Population .............................................................................................21 
4.3.2.1. 
Inclusion Criteria, Symptomatic Population...............................................................21 
4.3.2.2. 
Exclusion Criteria, Symptomatic Population .............................................................22 
4.3.3. 
Reasons for Withdrawal .............................................................................................23 
4.4. 
Treatments ..................................................................................................................23 
4.4.1. 
Treatments Administered ............................................................................................23 
4.4.2. 
Method of Assigning Subjects to Treatment Groups (Randomization) .....................24 
4.4.3. 
Blinding ......................................................................................................................25 
4.5. 
Efficacy and Safety Variables ....................................................................................25 


Statistical Analysis Plan - DMID Protocol: 15-0015  
Version 2.0 
29 April 2020 
Table of Contents (continued) 
 
- iv - 
PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
4.5.1. 
Efficacy Variables ......................................................................................................25 
5. 
SAMPLE SIZE CONSIDERATIONS .......................................................................30 
6. 
GENERAL STATISTICAL CONSIDERATIONS ....................................................31 
6.1. 
General Principles .......................................................................................................31 
6.2. 
Timing of Analyses .....................................................................................................31 
6.3. 
Analysis Populations ..................................................................................................31 
6.3.1. 
Modified Intention-to-Treat (mITT) Population ........................................................31 
6.3.1.1. 
Exclusions Due to Violation of Eligibility Criteria ....................................................32 
6.3.2. 
Per Protocol Population ..............................................................................................32 
6.3.3. 
Safety Population ........................................................................................................32 
6.4. 
Covariates and Subgroups ..........................................................................................32 
6.5. 
Missing Data ...............................................................................................................33 
6.5.1. 
Sensitivity Analyses ....................................................................................................34 
6.6. 
Interim Analyses and Data Monitoring ......................................................................34 
6.7. 
Multiple Comparisons/Multiplicity ............................................................................35 
7. 
STUDY SUBJECTS ...................................................................................................36 
7.1. 
Disposition of Subjects ...............................................................................................36 
7.2. 
Protocol Deviations ....................................................................................................36 
8. 
EFFICACY EVALUATION ......................................................................................37 
8.1. 
Primary Efficacy Analysis ..........................................................................................37 
8.1.1. 
Amebiasis Group ........................................................................................................37 
8.1.2. 
Giardiasis Group .........................................................................................................38 
8.2. 
Secondary Efficacy Analyses .....................................................................................38 
8.2.1. 
Amebiasis Group ........................................................................................................38 
8.2.2. 
Giardiasis Group .........................................................................................................41 
9. 
SAFETY EVALUATION ..........................................................................................42 
9.1. 
Demographic and Other Baseline Characteristics ......................................................42 
9.1.1. 
Prior and Concurrent Medical Conditions ..................................................................42 
9.1.2. 
Prior and Concomitant Medications ...........................................................................42 
9.2. 
Measurements of Treatment Compliance ...................................................................42 
9.3. 
Adverse Events ...........................................................................................................42 


Statistical Analysis Plan - DMID Protocol: 15-0015  
Version 2.0 
29 April 2020 
Table of Contents (continued) 
 
- v - 
PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
9.3.1. 
Solicited Events and Symptoms .................................................................................42 
9.3.2. 
Unsolicited Adverse Events ........................................................................................43 
9.4. 
Deaths, Serious Adverse Events and other Significant Adverse Events ....................44 
9.5. 
Pregnancies .................................................................................................................44 
9.6. 
Clinical Laboratory Evaluations .................................................................................44 
9.7. 
Vital Signs and Physical Evaluations .........................................................................44 
9.8. 
Concomitant Medications ...........................................................................................45 
10. 
PHARMACOKINETICS ...........................................................................................46 
11. 
IMMUNOGENICITY ................................................................................................47 
12. 
OTHER ANALYSES .................................................................................................48 
13. 
REPORTING CONVENTIONS ................................................................................49 
14. 
TECHNICAL DETAILS ............................................................................................50 
15. 
SUMMARY OF CHANGES IN THE CONDUCT OF THE STUDY OR 
PLANNED ANALYSES ............................................................................................51 
16. 
REFERENCES ...........................................................................................................52 
17. 
LISTING OF TABLES, FIGURES, AND LISTINGS ..............................................53 
APPENDICES ...............................................................................................................................54 
APPENDIX 1. TABLE MOCK-UPS..........................................................................................55 
APPENDIX 2. FIGURE MOCK-UPS ......................................................................................123 
APPENDIX 3. LISTINGS MOCK-UPS ...................................................................................149 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
Version 2.0 
29 April 2020 
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PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
LIST OF ABBREVIATIONS 
AE 
Adverse Event 
ALT 
Alanine Aminotransferase 
AST 
Aspartate Aminotransferase 
BP 
Blood Pressure  
BUN 
Blood Urea Nitrogen 
C 
Celsius 
CI 
Confidence Interval 
CRF 
Case Report Form 
DMID 
Division of Microbiology and Infectious Diseases 
DSMB 
Data and Safety Monitoring Board 
EDC 
Electronic Data Capture 
ELISA 
Enzyme-linked Immunosorbent Assay 
ER 
Emergency Room 
F 
Fahrenheit 
GGT 
Gamma Glutamyl Transferase 
GMT 
Geometric Mean Titer 
GMFR 
Geometric Mean Fold Rise 
icddr,b 
International Center for Diarrheal Disease Research, Bangladesh 
ICH 
International Conference on Harmonisation 
IRB 
Institutional Review Board 
ITT 
Intention to Treat 
L 
Liter 
LLN 
Lower Limit of Normal 
mcg 
Microgram 
MedDRA 
Medical Dictionary for Regulatory Activities  
mEq 
Milliequivalent 
mg 
Milligram 
mITT 
Modified Intention to Treat 
mL 
Milliliter 
MAR 
Missing at Random 


Statistical Analysis Plan - DMID Protocol: 15-0015 
Version 2.0 
29 April 2020 
List of Abbreviations (continued) 
 
- vii - 
PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
MCAR 
Missing Completely at Random 
MNAR 
Missing Not at Random 
N 
Number (typically refers to subjects) 
NIH 
National Institutes of Health 
PI 
Principal Investigator 
PP 
Per Protocol 
PT 
Preferred Term 
RBC 
Red Blood Cell 
RCD 
Reverse Cumulative Distribution  
SAE 
Serious Adverse Event 
SD 
Standard Deviation 
SDCC 
Statistical and Data Coordinating Center 
SMC 
Safety Monitoring Committee 
SOC  
System Organ Class 
SOP 
Standard Operating Procedures 
U 
Units 
ULN 
Upper Limit of Normal 
WBC 
White Blood Cell 
WHO 
World Health Organization 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
Version 2.0 
29 April 2020 
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PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
1. 
PREFACE 
The Statistical Analysis Plan (SAP) for “Phase IIa Randomized, Single-blinded, Placebo-controlled Clinical 
Trial of the Reprofiled Drug Auranofin for GI Protozoa” (DMID Protocol 15-0015) describes and expands 
upon the statistical information presented in the protocol. 
This document describes all planned analyses and provides reasons and justifications for these analyses. It 
also includes sample tables, listings, and figures planned for the final analyses. Regarding the final analyses 
and Clinical Study Report (CSR), this SAP follows the International Conference on Harmonisation of 
Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) Guidelines, as indicated in 
Topic E3 (Structure and Content of Clinical Study Reports), and more generally is consistent with Topic E8 
(General Considerations for Clinical Trials) and Topic E9 (Statistical Principles for Clinical Trials). The 
structure and content of the SAP provides sufficient detail to meet the requirements identified by the FDA and 
ICH, while all work planned and reported for this SAP will follow internationally accepted guidelines 
published by the American Statistical Association and the Royal Statistical Society for statistical practice.  
This document contains four sections: (1) a review of the study design, (2) general statistical considerations, 
(3) comprehensive statistical analysis methods for efficacy and safety outcomes, and (4) a list of proposed 
tables and figures. Within the table, figure, and listing mock-ups (Appendices 1, 2, and 3), references to CSR 
sections are included. Any deviation from this SAP will be described and justified in protocol amendments 
and/or in the CSR, as appropriate. The reader of this SAP is encouraged to also review the study protocol for 
details on conduct of the study and the operational aspects of clinical assessments.   


Statistical Analysis Plan - DMID Protocol: 15-0015  
Version 2.0 
29 April 2020 
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PRIVILEGED AND CONFIDENTIAL COMMUNICATION 
2. 
INTRODUCTION 
This study investigates the effectiveness of auranofin for two gastrointestinal protozoa: E. histolytica and 
Giardia.  This proposed clinical trial follows from key findings in studies supported by U01 AI0778822 
entitled, “Novel Therapeutics for Class B Protozoa,” to design and develop highly effective antiparasitic 
agents for Class B Protozoa. The development of the first high-throughput whole cell screens for E. 
histolytica (Debnath, 2012) and G. lamblia (Gut et al., 2011) against FDA-approved drugs and bioactive 
compounds (a “repurposing” screen), found that auranofin had a ten-fold lower IC50 (0.5 µM) compared with 
metronidazole (5.2 µM) for E. histolytica and an equivalent IC50 for Giardia  (4.0 µM). Auranofin is an orally 
available gold-containing compound that has been in clinical use in the treatment of rheumatoid arthritis for 
25 years.  As previous research has found auranofin to be the first drug active against Entamoeba and Giardia 
with a clearly defined target, the FDA has given auranofin Orphan Drug status. 
This study is a Phase IIa clinical trial conducted in Bangladesh. The initial plan for this study included a 
single site, the International Center for Diarrheal Disease Research, Bangladesh (icddr,b).  The study initially 
planned to enroll 136 males and non-pregnant females aged 18 to 65 years with asymptomatic amebiasis (68 
subjects) or giardiasis (68 subjects).  Participants within each disease group were to be randomized in a 1:1 
ratio to receive auranofin or a placebo.  The amebiasis group was to receive 6 mg auranofin (or placebo) daily 
for seven days while the giardiasis group was to receive 6 mg auranofin (or placebo) daily for five days.  The 
initial primary goal of the study, reflected in versions 1 - 4 of the protocol, was to compare the proportion of 
subjects with parasitological response, defined as no detection of cysts or trophozoites on microscopic exam 
or negative antigen detection, between treatment arms by Day 7 for amebiasis and by Day 5 for giardiasis.   
However, the protocol endpoints and study population were revised because after screening 2,396 people 
during 17 months with only 34 enrolling into the study, it became apparent that enrolling 136 asymptomatic 
subjects would be infeasible.  For this reason, eligibility criteria were revised, starting with protocol version 5, 
to enroll symptomatic rather than asymptomatic adult patients (still with amebiasis or giardiasis).  The goal, as 
before, was to enroll 68 in each disease group and randomize in a 1:1 ratio within each disease group to 
receive either auranofin or placebo.  The dosing regimen was unchanged, but the primary outcome was 
revised for the new (symptomatic) study population.  The new primary outcome is resolution of diarrhea (less 
than 3 loose stools / 24 hours) by Day 7 for the symptomatic amebiasis group and by Day 5 for the 
symptomatic giardiasis group.  Furthermore, in version 7 of the protocol, a new site was also added (Rajshahi 
Medical College Hospital), after which randomization was stratified by site as well as disease group.  The 
protocol revision specified that the asymptomatic and symptomatic subjects would be analyzed separately. 
The study populations, study design, and primary outcome for each of these populations are summarized by 
protocol version in Table 1: 
 
 


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Table 1: 
Summary of Study Population, Study Design, and Primary Outcome by Protocol Version 
Protocol 
Versions 
Study 
Population 
Target 
Sample 
Size 
Treatment 
Allocation 
Dosing 
Regimen 
Primary Outcome 
1-4 
Asymptomatic 
amebiasis 
68  
1:1 (auranofin: 
placebo) 
7 days 
Parasitological response (no 
detection of cysts or trophozoites 
on microscopic exam or negative 
antigen detection) by Day 7 
Asymptomatic 
giardiasis 
68 
1:1 (auranofin: 
placebo) 
5 days  
Parasitological response (no 
detection of cysts or trophozoites 
on microscopic exam or negative 
antigen detection) by Day 5 
5-7 
Symptomatic 
amebiasis 
68 
1:1 (auranofin: 
placebo) a 
7 days 
Resolution of diarrhea (less than 3 
loose stools / 24 hours) by Day 7 
Symptomatic 
giardiasis 
68 
1:1 (auranofin: 
placebo) a 
5 days  
Resolution of diarrhea (less than 3 
loose stools / 24 hours) by Day 5 
a Randomization was also stratified by site for subjects enrolled under protocol version 7 because a second site was added in that 
protocol version. 
Secondary objectives for the asymptomatic population are summarized in Table 2, while those for the 
symptomatic population are summarized in Table 3.  The changes in secondary outcomes for the symptomatic 
population are shown in blue in Table 3. 
Table 2: 
Secondary Outcomes for the Asymptomatic Study Population 
Outcome 
Time points,  
Asymptomatic Amebiasis 
Group 
Time points, 
 Asymptomatic Giardiasis 
Group 
Parasitological response (no detection of cysts or 
trophozoites on microscopic exam or negative antigen 
detection) 
Days 3 and 5 
Day 3 
Rate of decrease in trophozoite/cyst load by qPCR in 
stools 
Days 3, 5, and 7 
Days 3 and 5 
Negative stool antigen test 
Days 3, 5, 7, and 14 
N/A 
Sustained cure (no detection of cysts or trophozoites by 
microscopic exam or negative antigen detection) 
Days 14 and 28 
Days 14 and 28 
Relapse (same strain) or re-infection (new strain) with 
positive stools by genotyping the initial vs. subsequent 
strain.   
Day 14 and/or 28 
Day 14 and/or 28 
 
 


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Table 3: 
Secondary Outcomes for the Symptomatic Study Population 
Changes to outcomes from the asymptomatic population are shown in blue: 
Outcome 
Time points,  
Symptomatic Amebiasis 
group 
Time points, 
 Symptomatic Giardiasis 
group 
Parasitological response (no detection of trophozoites 
on microscopic exam a) 
 
 
Days 3, 5, and 7 b 
Day 3 and 5 b 
Rate of decrease in trophozoite/cyst load by qPCR in 
stools 
Days 3, 5, and 7 
Days 3 and 5 
Negative stool antigen test 
Days 3, 5, 7, and 14 
Days 3 and 5 
Sustained cure (no detection of trophozoites by 
microscopic exam) c 
Days 14 and 28 
Days 14 and 28 
Relapse (same strain) or re-infection (new strain) with 
positive stools by genotyping the initial vs. subsequent 
strain.   
Day 14 and/or 28 
Day 14 and/or 28 
Time to resolution of diarrhea d 
Assessment over the study 
period 
Assessment over the study 
period 
a The definition of parasitological response was revised to allow for cysts and remove the antigen test. 
b Moved from primary to secondary outcome.   
c Modified so definition of “cure” matches the updated definition of parasitological response for this population. 
d New outcome for this study population. 
 


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2.1. 
Purpose of the Analyses 
The overall goal is to assess the safety and efficacy of auranofin compared to a placebo for the treatment of 
asymptomatic amebiasis and giardiasis among males and non-pregnant females aged 18-65 years old in 
Bangladesh in two study populations: asymptomatic and symptomatic. 
Two analyses will be conducted – one for the asymptomatic patients and another for the symptomatic 
patients, each according to their corresponding outcome measures.  This SAP includes analysis plans for both 
study populations. 


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3. 
STUDY OBJECTIVES AND ENDPOINTS 
3.1. 
Study Objectives 
Auranofin is a “repurposed” drug with established pharmacology and is approved for human use by the FDA.  
This study assesses clinical efficacy for two new indications, amebiasis and giardiasis, with a randomized 
Phase IIa, placebo-controlled, single-blinded, superiority trial comparing auranofin for treatment of amebiasis 
and giardiasis to placebo. Parasite clearance will be analyzed in the asymptomatic population, while 
resolution of diarrhea will be studied in the symptomatic population. Ultimately, the goal will be to expand 
the trial to include symptomatic children after proving efficacy and safety in adults. 
All subjects will have the same screening protocol, but the trial will evaluate subjects with amebiasis or 
giardiasis with separate primary and secondary endpoints.  Below, objectives for the asymptomatic population 
are excerpted below from Version 4.0 of the Study Protocol, the first version under which participants were 
enrolled, and those for the symptomatic population are excerpted below from Version 7.0 of the Study 
Protocol. 
3.1.1. 
Objectives for the Asymptomatic Population 
Primary E. histolytica: 
To compare the proportion of subjects with parasitological response (no detection of cysts or trophozoites of 
E. histolytica on microscopic exam or negative antigen detection) by Day 7  
Secondary E. histolytica: 
1. To compare the proportion of subjects with parasitological response (no detection of cysts or 
trophozoites on microscopic exam or negative antigen detection) by Day 3 and 5 
2. To compare the rate of decrease in trophozoite/cyst load by qPCR in stools by Days 3, 5, and 7 
3. To compare the proportion of subjects with negative stool antigen test by Days 3, 5, 7, and 14 
4. To compare the proportion of subjects with sustained cure (no detection of cysts or trophozoites by 
microscopic exam or negative antigen detection) at 14 and 28 days 
5. To compare the proportion of subjects with relapse (same strain) or re-infection (new strain) with 
positive stools at 14 and/or 28 days by genotyping the initial vs. subsequent strain 
Primary Giardia: 
To compare the proportion of subjects with parasitological response (no detection of cysts or trophozoites on 
microscopic exam or negative antigen detection) by Day 5 
Secondary Giardia: 
1. To compare the proportion of subjects with parasitological response (no detection of cysts or 
trophozoites on microscopic exam or negative antigen detection) by Day 3  
2. To compare the rate of decrease in trophozoite/cyst load by qPCR in stools by Days 3 and 5 
3. To compare the proportion of subjects with sustained cure (no detection of cysts or trophozoites by 
microscopic exam or negative antigen detection) at 14 and 28 days 


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4. To compare the proportion of subjects with relapse (same strain) or re-infection (new strain) with 
positive stools at 14 and/or 28 days by genotyping the initial vs. subsequent strain 
3.1.2. 
Objectives for the Symptomatic Population 
Primary E. histolytica: 
To compare the proportion of subjects with stools positive by rapid EIA and positive antigen detection EIA 
for E. histolytica at enrollment with resolution of diarrhea (less than 3 loose stools/ 24 hours) by Day 7.  
Secondary E. histolytica: 
• To compare the proportion of subjects with positive rapid EIA and positive antigen detection EIA 
for E. histolytica and trophozoites on smear (wet mount or concentrated trichrome) at enrollment 
with parasitological response (no detection of trophozoites on microscopic exam) by Day 7.  
• To compare the proportion of subjects with positive rapid EIA and positive antigen detection EIA 
for E. histolytica and trophozoites on smear at enrollment with parasitological response (no 
detection of trophozoites on microscopic exam) by Day 3 and 5. 
• To compare the rate of decrease in trophozoite/cyst load by qPCR in stools by Days 3, 5, and 7. 
• To compare the proportion of subjects with negative stool antigen test by Days 3, 5, 7, and 14. 
• To compare the proportion of subjects with sustained cure (no detection of trophozoites by 
microscopic exam) at 14 and 28 days. 
• To compare the proportion of subjects with relapse (same strain) or re-infection (new strain) with 
positive stools at 14 and/or 28 days by genotyping the initial vs. subsequent strain. 
• To compare the time to resolution of diarrhea (less than 3 loose stools/24 hours) 
Primary Giardia: 
To compare the proportion of subjects with stool positive rapid EIA and positive antigen detection EIA for 
Giardia at enrollment with resolution of diarrhea (less than 3 loose stools/24 hours) by Day 5. 
Secondary Giardia: 
• To compare the proportion of subjects with parasitological response (no detection of trophozoites 
on microscopic exam) by Day 3 and 5. 
• To compare the rate of decrease in trophozoite/cyst load by qPCR in stools by Days 3 and 5. 
• To compare the proportion of subjects with negative stool antigens by days 3 and 5. 
• To compare the proportion of subjects with sustained cure (no detection of trophozoites by 
microscopic exam) at 14 and 28 days. 
• To compare the proportion of subjects with relapse (same strain) or re-infection (new strain) with 
positive stools at 14 and/or 28 days by genotyping the initial vs. subsequent strain. 
• To compare the time to resolution of diarrhea (less than 3 loose stools/24 hours) 


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3.2. 
Endpoints 
The asymptomatic endpoints are excerpted below from the Study Protocol, Version 4.0 (the first version 
under which participants were enrolled), and the symptomatic endpoints are excerpted from the Study 
Protocol, Version 7.0. 
3.2.1. 
Asymptomatic Population Endpoints 
Primary Outcome Measures 
1. Proportion of subjects with parasitological response of E. histolytica (no detection of cysts or 
trophozoites of E. histolytica on microscopic exam or negative antigen detection) by Day 7. 
2. Proportion of subjects with parasitological response of Giardia (no detection of cysts or trophozoites 
on microscopic exam or negative antigen detection) by Day 5   
Secondary Outcome Measures 
1. E. histolytica: 
a. Proportion of subjects with parasitological response (no detection of cysts or trophozoites on 
microscopic exam or negative antigen detection) by Day 3 and 5 
b. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3, 5, and 7 
c. Proportion of subjects with negative stool antigen test by Days 3, 5, 7, and 14 
d. Proportion of subjects with sustained cure (no detection of cysts or trophozoites by microscopic 
exam or negative antigen detection) at 14 and 28 days  
e. Proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 
14 and 28 days by genotyping the initial vs. subsequent strain  
2. Giardia: 
a. Proportion of subjects with parasitological response (no detection of cysts or trophozoites on 
microscopic exam or negative antigen detection) by Day 3  
b. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3 and 5 
c. Proportion of subjects with sustained cure (no detection of cysts or trophozoites by microscopic 
exam or negative antigen detection) at 14 and 28 days  
d. Proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 
14 and 28 days by genotyping the initial vs. subsequent strain 
3.2.2. 
Symptomatic Population Endpoints 
Primary Outcome Measures 
1. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for E. histolytica 
and resolution of diarrhea (less than 3 loose stools/ 24 hours) by Day 7.  
2. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for Giardia and 
resolution of diarrhea (less than 3 loose stools/24 hours) by Day 5.  
Secondary Outcome Measures 
1. E. histolytica: 


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a. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for E. histolytica 
and trophozoites on smear at enrollment with parasitological response (no detection of 
trophozoites on microscopic exam) by Day 7. 
b. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for E. histolytica 
and trophozoites on smear at enrollment with parasitological response (no detection of 
trophozoites on microscopic exam) by Day 3 and 5. 
c. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3, 5, and 7 
d. Proportion of subjects with negative stool antigen test by Days 3, 5, 7, and 14 
e. Proportion of subjects with sustained cure (no detection of trophozoites by microscopic exam) at 
14 and 28 days  
f. Proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 
14 and 28 days by genotyping the initial vs. subsequent strain 
g. Time to resolution of diarrhea (less than 3 loose stools/24 hours) 
2. Giardia: 
a. Proportion of subjects with positive rapid EIA and positive antigen detection EIA for Giardia and 
trophozoites on smear at enrollment with parasitological response (no detection of trophozoites on 
microscopic exam) by Day 3 and 5  
b. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3 and 5 
c. Proportion of subjects with negative stool antigens by days 3 and 5.  
d. Proportion of subjects with sustained cure (no detection of trophozoites by microscopic exam) at 
14 and 28 days  
e. Proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 
14 and 28 days by genotyping the initial vs. subsequent strain 
f. Time to resolution of diarrhea (less than 3 loose stools/24 hours) 
3.3. 
Study Definitions and Derived Variables  
For the purposes of enrollment into the asymptomatic population, potential subjects are diagnosed with 
amebiasis or giardiasis if they have a positive TechLab antigen test, positive wet mount or trichome, and 
positive stool PCR for E. histolytica or Giardia.  In order to enroll, the subject must receive the diagnosis of 
amebiasis or giardiasis within 72 hours of enrollment.  If a subject is infected with both E. histolytica and 
Giardia, they will be enrolled in the E. histolytica study arm. Once the E. histolytica study arm is fully 
enrolled, any subsequent dual infected subjects will be enrolled in the Giardia arm.  
For enrollment into the symptomatic population, potential subjects are diagnosed with amebiasis or 
giardiasis if they have a positive rapid EIA and positive antigen detection EIA of E. histolytica or Giardia.  
These subjects must also have diarrhea (3 or more loose stools within 24 hours of enrollment).  In order to 
enroll, the subject must receive the diagnosis of amebiasis or giardiasis within 4 days prior to enrollment.  If a 
subject is infected with both E. histolytica and Giardia, they will be enrolled in the E. histolytica study arm. 
Once the E. histolytica study arm is fully enrolled, any subsequent dual infected subjects will be enrolled in 
the Giardia arm.  If a subject is infected with both Giardia and Cryptosporidium, they will not be enrolled. 
Parasitological response is defined as no detection of cysts or trophozoites on microscopic exam or negative 
antigen detection, for the asymptomatic population.  Both tests will be performed on each subject.  For the 
symptomatic population, however, parasitological response is defined as no detection of trophozoites on 
microscopic exam.  The microscopic exam may include both wet mount and trichrome staining.  If both were 
performed, then both must be negative for the microscopic exam to be negative.  


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Sustained cure on Day 14 is defined, for the asymptomatic amebiasis group, to be no detection of cysts or 
trophozoites on microscopic exam or negative antigen detection on Day 7 and again on Day 14.  Sustained 
cure on Day 14 is defined, for the symptomatic amebiasis group, to be no detection of trophozoites on 
microscopic exam on Day 7 and again on Day 14.  Sustained cure on Day 14 is defined analogously for the 
giardiasis groups but is based on measurements taken on Day 5 and 14.   
Sustained cure on Day 28 is defined for the asymptomatic amebiasis group to be no detection of cysts or 
trophozoites on microscopic exam or negative antigen detection on Days 7, 14, and 28 (if specimens are 
obtained at each time point).   Sustained cure on Day 28 is defined for the symptomatic amebiasis group to 
be no detection of trophozoites on microscopic exam on Days 7, 14, and 28 (if specimens are obtained at each 
time point).   It is defined analogously for the giardiasis groups but is based on measurements taken on Day 5, 
14, and 28.   
Relapse at Day X is defined as having a positive stool PCR at day X that is identified via genotyping to be 
identical to the strain of the original infection.   
Reinfection at Day X is defined as having a positive stool PCR at day X that is identified via genotyping to be 
non-identical to the strain of the original infection.   
In the two above definitions, the original infection is defined to be the strain of parasite measured in the stool 
sample collected on Visit 01. 


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4. 
INVESTIGATIONAL PLAN 
4.1. 
Overall Study Design and Plan 
This 28-day, Phase IIa, two-arm, randomized, placebo-controlled, single-blinded, superiority treatment study 
in 136 males and non-pregnant females ≥ 18 to 65 years of age was designed to compare placebo to once 
daily doses of auranofin (Ridaura) for adults with asymptomatic amebiasis (68 subjects) or giardiasis (68 
subjects).  Potential adult subjects who are otherwise healthy from the single site of the Mirpur community of 
Bangladesh were chosen from a randomized census list for a visit by a health professional from icddr,b. Once 
eligibility and consent were confirmed, subjects were stratified by infection (a 2-level variable of either 
amebiasis vs. giardiasis), and then within each stratum, randomized 1:1 to receive auranofin or placebo. 
Because of difficulties in enrolling the target number of participants under the original design, the eligibility 
criteria were revised to study symptomatic rather than asymptomatic participants, beginning with protocol 
version 5.  The new goal is to enroll a total of 136 symptomatic participants (68 in each disease group), in 
addition to the asymptomatic participants.  The asymptomatic participants who enrolled under protocol 
versions 1-4 will be analyzed separately.  In addition, a new site was added in protocol version 7.  
Both study populations (asymptomatic and symptomatic) have the same dosing regimen.  Subjects in the 
amebiasis group receive 6 mg auranofin or placebo delivered orally on Days 1, 2, 3, 4, 5, 6, and 7; those in the 
giardiasis group receive 6 mg auranofin or placebo orally on Days 1, 2, 3, 4, and 5.  For both study 
populations, follow-up visits are conducted on dosing days, as well as on Day 7, 14, and 28.  For the 
symptomatic population only, any subject with worsening diarrhea as assessed by the study physician, may 
have their dose discontinued and receive standard of care metronidazole.  In this population, subjects who 
remain stool positive for giardiasis or amebiasis on the Day 28 visit will have a Day 30 visit to receive the 
standard of care treatment for giardiasis or amebiasis.  
The placebo capsules are similar, but not identical to the study drug, so this study is considered single-blind. 
The laboratory personnel performing evaluations are blinded to treatment assignment.  Subjects, investigators, 
and study personnel performing any study-related assessments following study product administration are not 
to be informed of group allocation, but could detect differences in study and placebo capsules. They were 
instructed to perform their clinical or AE assessments as though blinded to prevent potential bias. 
The planned duration of subject participation for the asymptomatic population was 42 days of face to face 
visits, including the pre-enrollment screening period of 14 days.  For the symptomatic population, it is 34 days 
of visits, including the pre-enrollment screening period of 4 days.  It is anticipated that it will take 
approximately 3.5 years to finish the study.  The Data Safety Monitoring Board (DSMB) meets at least 
annually to review safety data.  Study enrollment will be stopped for DSMB review of safety data when the 
study reaches 50% enrollment (i.e. 68 subjects enrolled).  DSMB will review safety data after the 68th subject 
completes visit 8.  Data review for efficacy will be performed at the completion of enrollment. The final 
review meeting will occur 6 to 8 months after clinical database lock to review the cumulative unblinded safety 
and efficacy data for the study.   
The schematics in Figure 1 and Figure 2 visualize the study schedule for the symptomatic population.  The 
study schedule for the asymptomatic population is the same except that it excludes the Day 30 visit and has 
four screening visits instead of two. 


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4.2. 
Discussion of Study Design, Including the Choice of Control Groups 
This study compares auranofin (6 mg daily) given for 7 days to subjects infected with amebiasis or 5 days for 
those infected with giardiasis, to a placebo. 
The study is single-blind because the auranofin capsules are similar, but not identical, to the placebo capsules.  
Each auranofin dose is comprised of two capsules, each containing 3 mg of white crystalline powder.  Each 
capsule has an opaque brown cap and opaque tan body and is imprinted with the produce name RIDAURA.  
The placebo capsules are similar to auranofin capsules with opaque brown caps and an opaque tan body but 
are not imprinted with the product name RIDAURA.  Since the treatment and placebo capsules are not 
identical, patients and staff dispensing medication are not considered blind.  The subjects, investigators, and 
study personnel performing any study-related assessments following study product administration are not to 
be informed of group allocation but could detect differences in study and placebo capsules. They were 
instructed to perform their clinical or AE assessments as though blinded to prevent potential bias.  However, 
the laboratory personnel performing evaluations are blinded to treatment assignment.  
The selected dose of 6 mg per day of auranofin is the standard maintenance dose for rheumatoid arthritis.  In 
the case of rheumatoid arthritis, auranofin may be prescribed for many months.  Therefore, the dose selected 
for this study is consistent with current use of the medication and the dose intervals are shorter than current 
practice. 
4.3. 
Selection of Study Population 
4.3.1. 
Asymptomatic Population 
The inclusion and exclusion criteria for the asymptomatic population are excerpted below from the Study 
Protocol, Version 4.0, the earliest protocol version under which subjects were enrolled. 
4.3.1.1. 
Inclusion Criteria, Asymptomatic Population 
1. Provide written informed consent prior to initiation of any study procedures. 
2. Able to understand and comply with planned study procedures and be available for all study visits. 
3. Male or non-pregnant female 18-65 years of age, inclusive. 
4. Amebiasis or giardiasis identified by positive TechLab antigen test, positive wet mount or trichome 
(confirmed by PCR), and positive stool PCR for E. histolytica or Giardia.1 
1If a subject is infected with both E. histolytica and Giardia, they will be enrolled in the E. histolytica study arm. 
Once the Entamoeba study arm is fully enrolled, any subsequent dual infected subjects will be enrolled in the Giardia 
arm.  
5. In otherwise good health2. 
2As determined by medical history and targeted physical examination, if indicated based on medical history, to 
evaluate acute or currently ongoing chronic medical diagnoses or conditions that would affect the assessment of 
eligibility and safety of subjects. Existing medical diagnoses or conditions (except those in the Subject Exclusion 
Criteria) must be deemed as stable chronic medical conditions. A stable chronic medical condition is defined as no 
change in prescription medication, dose, or frequency of medication in the last 3 months (90 days) and health 
outcomes of the specific disease are considered to be within acceptable limits in the last 6 months (180 days).  Any 
change due to change of health care provider, insurance company, or that is done for financial reasons, as long as in 
the same class of medication, will not be considered a violation of this inclusion criterion.  Any change in 
prescription medication due to improvement of a disease outcome, as determined by the site principal investigator or 


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appropriate sub-investigator, will not be considered a violation of this inclusion criterion. Subjects may be on chronic 
or as needed (prn) medications if, in the opinion of the site principal investigator or appropriate sub-investigator, they 
pose no additional risk to subject safety. Topical, nasal, and inhaled medications, vitamins, and contraceptives are 
permitted. 
6. Vital signs (oral temperature, pulse, and blood pressure) are all within normal protocol-defined ranges 
(abnormal criteria defined in Section 9.2.3). 
7. Laboratory tests (blood urea nitrogen, creatinine, AST, ALT, white blood cells, platelets, hemoglobin) 
are all within protocol-defined reference ranges3. 
3The normal ranges for Icddr,b laboratory tests include (a) blood urea nitrogen (BUN) between 5 and 24 mg/dL, 
inclusive, (b) creatinine between 53  and106 umol/L,  inclusive, males and between 44  and 97umol/L, inclusive, for 
females, (c) AST of 38 U/L or less for males and 32 U/L or less for females (d) ALT of 41.0 U/L or less for males and 
31 U/L or less for females,  (e) WBC between 4.0 and 11.0 109/L, inclusive (f) platelets between 150 and 450 109/L, 
inclusive, (g) hemoglobin between 12.5 and 17.5 gm/dL, inclusive, for males and 11.5 and 16.5 gm/dL, inclusive, for 
females,  
Subjects will be eligible for enrollment with the following laboratory values: 
• Blood urea nitrogen less than 30 mg/dL 
• Creatinine less than 133 umol/L 
• AST or ALT less than 60.0 U/L 
• White cell count between 3.5 and 11.0 (109/L) 
• Platelets between 100 and 500 (109/L) 
• Hemoglobin between 11.0 and 18.0 gm/dL 
8. Urinalysis with no greater than trace protein. If a high protein is confirmed to be due to menstruation, it 
should be repeated. 
9. Women of reproductive potential4 must have a negative urine pregnancy test within 72 hours of 
starting study medications.  
4Female subjects who are surgically sterile via tubal sterilization, bilateral oophorectomy or hysterectomy or who have 
been postmenopausal for greater than 1 year are not considered to be of reproductive potential. 
10. Female subjects participating in sexual activity that could lead to pregnancy must agree to use highly 
effective5 contraception while receiving auranofin and for 15 weeks after. 
5Highly effective methods of contraception are defined as having low failure rates (i.e. less than 1% per year) when 
used consistently and correctly and may include, but are not limited to, abstinence from intercourse, monogamous 
relationship with a vasectomized partner, male condoms with spermicide, diaphragm with spermicide, intrauterine 
devices, and licensed hormonal methods. If an eligible female of child-bearing potential is not using effective 
contraception, hormonal contraceptives will be supplied to female subjects and condoms will be provided for her 
partner while they are on the trial and for the follow-up period of 4 months total.  Females on effective forms of birth 
control will continue while on the study and for the follow-up period of 4 months total and also be provided condoms 
for their partners. The method and compliance of birth control used will be confirmed and documented at all study 
visits. 
4.3.1.2. 
Exclusion Criteria, Asymptomatic Population 
Subjects meeting any of the following exclusion criteria at baseline will be excluded from study participation 
within 1 week of the initial visit: 


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1. Known intolerance of auranofin or gold compounds. 
2. Pregnant or breastfeeding women or women who plan to become pregnant or breastfeed at any given 
time during the study or within 2 months of study completion. 
3. Diarrhea (> 3 loose stools/ 24 hours) within 7 days prior to starting study medications. 
4. Anticipated travel of more than 50 km from study site planned in next month. 
5. Current use of systemic antibiotics or metronidazole. 
6. Has any condition that would, in the opinion of the site investigator, place the subject at an 
unacceptable risk of injury or render the subject unable to meet the requirements of the protocol. 
7. Concurrent participation in other investigational protocols or receipt of an investigational product 
within the previous 30 days. 
8. History of alcohol or drug abuse within the last five years. 
4.3.2. 
Symptomatic Population 
The inclusion and exclusion criteria for the symptomatic population are excerpted below from the Study 
Protocol, Version 7.0. 
4.3.2.1. 
Inclusion Criteria, Symptomatic Population 
1. Provide written informed consent prior to initiation of any study procedures. 
2. Able to understand and comply with planned study procedures and be available for all study visits. 
3. Male or non-pregnant, non-lactating females 18-65 years of age, inclusive. Females of reproductive 
potential currently using effective contraceptive methods are eligible. 
4. Amebiasis or giardiasis identified by rapid EIA and positive antigen detection EIA of stool1  
1If a subject is infected with both E. histolytica and Giardia, they will be enrolled in the E. histolytica study arm. Once the 
Entamoeba study arm is fully enrolled, any subsequent dual infected subjects will be enrolled in the Giardia arm.  If a 
subject is infected with both Giardia and Cryptosporidium, they will not be enrolled. 
5. Has diarrhea (defined as three or more loose stools) in the past 24 hrs, but is assessed to be clinically 
stable and in otherwise good health2. 
2As determined by medical history and targeted physical examination, if indicated based on medical history, to evaluate 
acute or currently ongoing chronic medical diagnoses or conditions that would affect the assessment of eligibility and 
safety of subjects. Existing medical diagnoses or conditions (except those in the Subject Exclusion Criteria) must be 
deemed as stable chronic medical conditions. A stable chronic medical condition is defined as no change in prescription 
medication, dose, or frequency of medication in the last 3 months (90 days) and health outcomes of the specific disease are 
considered to be within acceptable limits in the last 6 months (180 days).  Any change due to change of health care 
provider, insurance company, or that is done for financial reasons, as long as in the same class of medication, will not be 
considered a violation of this inclusion criterion.  Any change in prescription medication due to improvement of a disease 
outcome, as determined by the site principal investigator or appropriate sub-investigator, will not be considered a violation 
of this inclusion criterion. Subjects may be on chronic or as needed (prn) medications if, in the opinion of the site principal 
investigator or appropriate sub-investigator, they pose no additional risk to subject safety. Topical, nasal, and inhaled 
medications, vitamins, and contraceptives are permitted. 
 
 


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6. Vital signs (oral temperature, pulse, and blood pressure) are all within normal protocol-defined ranges  
7. Laboratory tests (blood urea nitrogen, creatinine, AST, ALT, white blood cells, platelets, hemoglobin) 
are all within protocol-defined ranges. 
Subjects will be eligible for enrollment with the following laboratory values:  
• Blood urea nitrogen less than or equal to 30 mg/dL 
• Creatinine less than or equal to 133umol/L 
• AST or ALT less than or equal to 70.0 U/L 
• White cell count between 3.5 and 13.0 inclusive(109/L) 
• Platelets between 131 and 550 (109/L) inclusive 
• Hemoglobin between 11.0 and 18.0 gm/dL inclusive 
8. Urinalysis with no greater than trace protein. If a high protein is confirmed to be due to menstruation, it 
should be repeated. 
9. Women of reproductive potential4 must have a negative urine pregnancy test within 72 hours of 
starting study medications.  
4Female subjects who are surgically sterile via tubal sterilization, bilateral oophorectomy or hysterectomy, who have been 
postmenopausal for greater than 1 year are not considered to be of reproductive potential. 
10. Female subjects participating in sexual activity that could lead to pregnancy must be using and 
continue to use highly effective5 contraception for a total of 4 months after enrollment. 
5Highly effective methods of contraception are defined as having low failure rates (i.e. less than 1% per year) when used 
consistently and correctly and may include, but are not limited to, abstinence from intercourse, monogamous relationship 
with a vasectomized partner, male condoms with spermicide, diaphragm with spermicide, intrauterine devices, and 
licensed hormonal methods.  Females on effective forms of birth control will continue while on the study and for the 
follow-up period of 4 months total. The method and compliance of birth control used will be confirmed and documented at 
all study visits. 
4.3.2.2. 
Exclusion Criteria, Symptomatic Population 
Subjects meeting any of the following exclusion criteria at baseline will be excluded from study participation 
within 1 week of the initial visit: 
1. Known intolerance of auranofin or gold compounds. 
2. Pregnant or breastfeeding women or women of reproductive potential not using effective 
contraception or who plan to become pregnant or breastfeed at any given time during the study or 
within 3 months of study completion. 
3. Use of metronidazole within the past 7 days. 
4. Has any condition that would, in the opinion of the site investigator, place the subject at an 
unacceptable risk of injury or render the subject unable to meet the requirements of the protocol. 
5. Concurrent participation in other investigational protocols or receipt of an investigational product 
within the previous 30 days. 
6. History of alcohol or drug abuse within the last five years. 


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4.3.3. 
Reasons for Withdrawal 
Subjects may voluntarily withdraw their consent for study participation at any time and for any reason, 
without penalty. Subjects who have received study product, regardless of the number of doses received, or 
who developed an AE or SAE will be encouraged to remain in the study to be followed for safety purposes. 
A subject may withdraw or be withdrawn from this study for any of the following reasons: 
1. Medical disease or condition, or any new clinical findings for which continued participation, in the 
opinion of the site principal investigator or appropriate sub-investigator, would compromise the safety 
of the subject, or would interfere with the subject's successful completion of the study, or would 
interfere with the evaluation of responses. 
2. Subject no longer meets eligibility criteria. 
3. As deemed necessary by the site principal investigator or appropriate sub-investigator for 
noncompliance or other reasons. 
4. Subject withdrawal of consent. 
5. Subject lost to follow-up. 
6. Termination of the study. 
7. New information becomes available that makes further participation unsafe. 
8. A subject will be discontinued from further receipt of auranofin or placebo if: 
a. The subject misses more than one dose of drug or placebo.  
b. Any clinical adverse event, laboratory abnormality, intercurrent illness, other medical condition 
or situation occurs such that continued participation in the study would not be in the best interest 
of the subject. 
4.4. 
Treatments 
4.4.1. 
Treatments Administered  
Auranofin (2, 3, 4, 6-tetra-O-acetyl-1-thio-D-glucopyranosato-S) (triethyl-phosphine gold), is an orally 
available gold-containing compound with established pharmacology and is approved for human use by the 
FDA. It is available in an oral form as capsules called Ridaura, which contain 3 mg of auranofin per capsule. 
Auranofin 3 mg capsules are manufactured by Prometheus Laboratories, San Diego, CA.  Subjects assigned to 
receive auranofin receive daily doses of 6 mg (two capsules) for either 7 days (amebiasis group) or 5 days 
(giardiasis group). 
Each capsule of auranofin has an opaque brown cap and opaque tan body, contains 3 mg of white crystalline 
powder, and is imprinted with the product name RIDAURA.  Inactive ingredients consist of benzyl alcohol, 
cellulose, cetylpyridinium chloride, D&C Red No. 33, FD&C Blue No. 1, FD&C Red No. 40, FD&C Yellow 
No. 6, gelatin, lactose, magnesium stearate, povidone, sodium lauryl sulfate, sodium starch glycolate, starch, 
titanium dioxide and trace amounts of other inactive ingredients.  Auranofin 3 mg capsules are packaged in 
bottles of 60 count. 
The placebo capsules are similar to auranofin capsules with opaque brown caps and an opaque tan body, but 
are not imprinted with the product name RIDAURA. They are manufactured by a contract supplier through 


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Fisher BioServices.  In order to mitigate potential unblinding of subjects and staff dispensing medication, the 
pharmacist will dispense daily doses according to the randomization assignment into dark, amber, light-
protective bottles with secure cap.  Each bottle will contain a daily dose (two capsules), so subjects in the 
giardiasis group will have 5 bottles; those in the amebiasis group will have 7 bottles.  Each individual bottle 
containing the single dose will be labeled with barcode and with randomization code and subject ID (as per 
local SOP) and double-checked before dispensing (i.e., before leaving the pharmacy and going out to the 
Mirpur Clinic).  The containers used to store the capsules are tight and light-resistant and will be identical for 
the placebo and auranofin capsules. 
4.4.2. 
Method of Assigning Subjects to Treatment Groups (Randomization) 
Per International Conference on Harmonisation (ICH) guideline E6: Good Clinical Practice (GCP), screening 
records will be kept at the icddr, b site to document the reason why an individual was screened, but failed trial 
entry criteria.  The reasons why individuals failed screening will be recorded in the Statistical and Data 
Coordinating Center’s (SDCC) Advantage eClinical Electronic Data Capture System. 
In the asymptomatic population, eligible subjects were stratified by disease strata (amebiasis or giardiasis), 
and then within each stratum, randomized 1:1 to receive auranofin or placebo.  Enrollment was to continue in 
the other stratum after one stratum reached full enrollment. All subjects were to receive doses of the assigned 
medication or placebo delivered orally on Days 1, 2, 3, 4, 5, 6, and 7 for amebiasis and Days 1, 2, 3, 4, and 5 
for giardiasis. 
For the symptomatic population, randomization is stratified by site to prevent confounding that could arise 
from imbalance in treatment allocations between sites.  Because the relative pace of enrollment between the 
two sites is unknown, the number of participants to be enrolled at each site is not fixed in advance.  With this 
approach, the faster enrolling site may enroll more subjects, allowing enrollment to complete as quickly as 
possible.  Each site may enroll up to 68 subjects into each stratum, so it is possible that one stratum could be 
filled entirely with subjects from a single site.  With this flexible approach, treatment assignments may not be 
perfectly balanced (overall or within each site), but they will be approximately balanced.  With sample sizes 
of 68 in each stratum, the impact of a small imbalance on power is negligible.  Once eligibility and consent 
are confirmed, symptomatic subjects are stratified by site and infection (amebiasis or giardiasis), and then 
within each stratum, randomized 1:1 to receive auranofin or placebo.  Enrollment will continue in the other 
arm after one arm reaches full enrollment. All subjects receive the assigned medication or placebo delivered 
orally on Days 1, 2, 3, 4, 5, 6, and 7 for amebiasis and Days 1, 2, 3, 4, and 5 for giardiasis. 
The randomization codes were prepared by statisticians at the SDCC and included in the enrollment module 
for the trial.  SDCC staff generated the randomization lists for each study population and site and sent 
corresponding documentation to the study sites. The randomization lists include the randomization codes to 
serially assign study treatment to subjects as they are assigned to the study (i.e., randomize subjects to study 
arm by study identification number).  This table did not contain unblinded information; study staff were not 
able to see the study arm assignment for each study identification number.  
Eligible study subjects were to be randomized under single-blind conditions by study staff members after 
consent. Since the placebo is similar but not identical to the study medication, only the lab personnel are truly 
blind, however the participants were not to be informed whether they receive study product or placebo. The 
randomization numbers were to be recorded on the study CRFs. 


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A designated individual at each site (icddr,b and Rajshahi Medical College hospital) was provided with the 
treatment key, which links the treatment code to the actual treatment assignment.  This is kept in a secure 
place. 
4.4.3. 
Blinding 
The unblinded pharmacist at the site will refer to the electronic treatment key provided for the trial by the 
SDCC to determine the treatment assignments for the subjects.  The pharmacist will maintain a hard copy of 
the treatment key (provided by the SDCC) under locked/secured conditions and will follow the randomization 
code.  
As described in Section 4.4.1 above, the placebo and auranofin capsules are similar in appearance, but not 
perfectly identical.  They will be stored in identical containers.  Study staff administering them and study 
subjects will not be informed of their minor differences in appearance nor of the treatment assignments of 
subjects.  The treatment assignment codes used by staff administering the medication will not reveal the 
treatment assignments of subjects. In addition, the laboratory personnel performing evaluations will be 
blinded to treatment assignment. 
The protocol contains no explicit provisions for emergency unblinding.  According to DMID policy, the study 
medical monitor responds to requests for emergency unblinding, and instructs the SDCC to release treatment 
codes only if necessary, to ensure that the subject receives appropriate clinical care. 
4.5. 
Efficacy and Safety Variables 
4.5.1. 
Efficacy Variables 
Table 4 and Table 5 summarize the efficacy variables and time points for the asymptomatic and symptomatic 
populations, respectively.   
Table 4: 
 Efficacy Variables for the Asymptomatic Study Population 
Outcome 
Type 
 
Outcome 
Time points,  
Asymptomatic 
amebiasis group 
Time points, 
 Asymptomatic 
giardiasis group 
Primary 
Parasitological response (no detection of cysts or 
trophozoites on microscopic exam or negative antigen 
detection) 
Day 7 
Day 5 
Secondary 
Parasitological response (no detection of cysts or 
trophozoites on microscopic exam or negative antigen 
detection) 
Days 3 and 5 
Day 3 
Rate of decrease in trophozoite/cyst load by qPCR in 
stools 
Days 3, 5, and 7 
Days 3 and 5 
Negative stool antigen test 
Days 3, 5, 7, and 14 
N/A 
Sustained cure (no detection of cysts or trophozoites by 
microscopic exam or negative antigen detection) 
Days 14 and 28 
Days 14 and 28 
Relapse (same strain) or re-infection (new strain) with 
positive stools by genotyping the initial vs. subsequent 
strain.   
Day 14 and/or 28 
Day 14 and/or 28 


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Table 5: 
Efficacy Variables and Time Points for the Symptomatic Study Population 
Changes from outcomes for the asymptomatic population are shown in blue: 
Outcome 
Type 
Outcome 
Time points,  
Symptomatic 
amebiasis group 
Time points, 
 Symptomatic 
giardiasis group 
Primary 
Resolution of diarrhea a 
Day 7 
Day 5 
Secondary 
Parasitological response (no detection of trophozoites on 
microscopic exam b) 
Days 3, 5, and 7 c 
Day 3 and 5 c 
Rate of decrease in trophozoite/cyst load by qPCR in stools 
Days 3, 5, and 7 
Days 3 and 5 
Negative stool antigen test 
Days 3, 5, 7, and 
14 
Days 3 and 5 
Sustained cure (no detection of trophozoites by microscopic 
exam) d 
Days 14 and 28 
Days 14 and 28 
Relapse (same strain) or re-infection (new strain) with 
positive stools by genotyping the initial vs. subsequent strain.   Day 14 and/or 28 
Day 14 and/or 28 
Time to resolution of diarrhea 
Assessment over 
the study period  
Assessment over 
the study period 
a New outcome for the symptomatic population 
b The definition of parasitological response was revised to allow for cysts and remove the antigen test.   
c Moved from primary to secondary outcome. 
d The definition of “cure” was updated to match the updated definition of “parasitological response”.   
Diarrhea will be assessed by self-report at each study visit. All other primary and secondary outcomes for 
both disease groups are measured on stool samples.  Subjects will provide stool samples on Days 1, 3, 5, 
7(window +1 day), 14 (window +3 days) and 28 (window +3 days).  The specimen will be stored in a cooler 
with ice pack for transport to the laboratory. All laboratory personnel will be blinded, and the specimens de-
identified.  
a. Testing of stool samples will be performed by the icddr,b  Parasitology Laboratory or Rajshahi 
Hospital laboratory and will include: 
i. Wet mount for ova and parasite examination; 
ii. TechLab EIA to detect E. histolytica and Giardia to identify protozoal infections; 
iii. PVA fixation for concentration and trichrome staining for ova and parasite examination 
iv. Qualitative PCR to detect E. histolytica and Giardia 
b. Off-site testing of stool samples at UCSD will include quantitative E. histolytica or Giardia testing 
performed at UCSD on shipped aliquots of DNA extracted from positive stools at icddr, b.  De-
identified DNA specimens labeled with the subject code will be obtained from stools on Days 1, 3, 
5, 7, 14, and 28 following enrollment. Quantitative PCR assays have been developed based on the 
small subunit rRNA gene, which is highly conserved among E. histolytica (Taniuchi, 2011) and 
Giardia isolates (Haque, 2007). 
Safety variables include solicited adverse events, vital signs, and clinical labs.  These are the same for the two 
disease groups and study populations.  The only exception is that although diarrhea is recorded for both 
populations, it is considered an efficacy rather than safety variable for the symptomatic population, because it 
is an eligibility criterion and an efficacy endpoint.  In other words, since everyone enrolls with diarrhea, their 


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subsequent diarrhea is probably not due to the treatment.  Solicited events and vital signs are recorded during 
planned visits until the Day 28 visit.  Vital signs are recorded during every planned follow-up visit.  
Solicited events are AEs that are known to occur with this study product.  Adverse event data is collected at 
each visit using specific questions and/or targeted physical examination. Based on the adverse reactions listed 
on the package insert, these include: loose stools or diarrhea, abdominal pain, nausea with or without 
vomiting, constipation, anorexia, flatulence, dyspepsia, dysgeusia, pruritus, hair loss, urticarial, stomatitis, 
conjunctivitis, glossitis, hematuria, and rashes. As noted above, diarrhea is considered an adverse event only 
in the asymptomatic population as it is a condition for enrollment in the symptomatic population.  Physical 
examinations look for abdominal pain, hair loss, urticaria, rashes, stomatitis, conjunctivitis, and glossitis.  In 
the symptomatic population, solicited AEs are not collected that are related to any prescribed metronidazole. 
Adverse event collection continues until Day 28 (following three half-lives of the drug).  Each solicited 
adverse event described above is graded as Mild (Grade 1) if it “Does not interfere with daily activity”, as 
Moderate (Grade 2) if it “Interferes with daily activity”, and as Severe (Grade 3) if it “Prevents daily activity 
(incapacitating)”.  Grading for vital signs, also collected on every visit, is given in Table 6.  Furthermore, 
hematology, biochemistry, and urine labs are analyzed on Study Day 7.  Grading for these adverse events is 
given in Table 7.  Some laboratory grading ranges were modified in protocol updates; these modifications are 
displayed in the table.  Laboratory results are graded according to the protocol effective on the date the 
sample was collected.  The effective date is the date that the site received IRB approval for the version and 
began implementing that version.  (The effective date is later than the protocol version date, which indicates 
when the version was approved by DMID.) The first subjects in the study enrolled under Protocol Version 
4.0, so laboratory ranges for earlier protocol versions are not displayed.  The effective dates are as follows: 
Version 4.0: 18Oct2016, Version 5.0: 12Dec2017, Version 6.0: 07Aug2018, Version 7.0: 23Jul2019.  
 
 


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Table 6: 
 Vital Signs Grading Scale 
Vital Signs* 
Vital Signs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Fever (°C) ** 
  
37.8-38.4 
 
38.5-38.9 
 
>= 39.0 
 
Hypertension 
(systolic) mm Hg 
141-150 
151-155 
>155 or 
an ER visit or hospitalization 
for malignant hypertension 
Hypertension 
(diastolic) mm Hg 
91-95 
96-100 
>100 or 
an ER visit or hospitalization 
for malignant hypertension 
Hypotension 
(systolic) mm Hg 
85-89 
80-84 
<80 or 
an ER visit or hospitalization 
for hypotensive shock 
Bradycardia – beats 
per minute *** 
50-54 
or >10 bpm less than 
baseline if baseline < 60 
45-49 
or >15 bpm less than baseline 
if baseline < 60 
<45 
or > 20 BPM or less than 
baseline if baseline < 60, 
or an ER visit or 
hospitalization for 
arrhythmia 
Tachycardia – beats 
per minute 
101-115 
116-130 
>130 
or an ER visit or 
hospitalization for 
arrhythmia 
* Subjects should be at rest for at least 15 minutes prior to vital sign measurements 
** Oral temperature; no recent hot or cold beverages or smoking 
*** When resting heart rate is between 60-100 beats per minute. Pulse and Blood Pressure assessed at Visit 1 (Day 1) will be 
considered baseline.  Sinus bradycardia among some healthy subject populations, for example, conditioned athletes maybe 
acceptable 
 
 


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Table 7: 
Laboratory Adverse Event Grading Scale 
NOTE:  
For grading ranges that were updated in protocol revisions, all ranges are shown in the below table, with 
their corresponding protocol versions.  No subjects were enrolled under Protocol Versions 1-3, so ranges from those 
versions are omitted.  Events are graded according to the protocol version that was active at the time of the event.  
Effective dates are: Version 4.0: 18Oct2016, Version 5.0: 12Dec2017, Version 6.0: 07Aug2018, Version 7.0: 23Jul2019 
Hematology 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
WBC 109/L (Decrease) 
Version 4: 3.0-3.9 
Versions 5-7: 3.0 – 3.4 
2.0-2.9 
<2.0 
WBC 109/L (Increase) 
Version 4: 11.1-13.0  
Versions 5-7: 13.1-13.9 
Version 4: 13.1-15.0  
Versions 5-7: 14.0-15.0 
>15.0  
Hgb g/dL (Decrease) (Female) 
Versions 4-6: 9-11.4 
Version 7: 9.1-10.9 
7.5-9.0 
<7.5 
Hgb g/dL (Decrease) (Male) 
Versions 4-6: 9-12.4 
Version 7: 9.1-10.9 
7.5-9.0 
<7.5 
Hgb g/dL (Increase) 
Versions 4-6: N/A 
Version 7: 18.1-18.5 
Versions 4-6: N/A 
Version 7: 18.6-19.0 
Versions 4-6: N/A 
Version 7: >19.0 
Platelets 109/L (Decrease) 
Version 4: 100-149 
Versions 5-7: 100-130  
50-99 
<50 
Platelets 109/L (Increase) 
Version 4: 451-480 
Versions 5-7: 551-600 
Version 4: 480-500 
Versions 5-7: 601-700 
Version 4: >500 
Versions 5-7: >700 
 
 
 
 
Chemistry 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
BUN mg/dL  
Version 4: 25-60 
Versions 5-7: 31-60  
Versions 4-7: 60-70† 
Version 8: >60 and ≤70 
>70 
Creatinine umol/L1 (Increase) 
(Male)  
Version 4: 107-176   
Versions 5-7: 134-176 
177-221 
>221  
Creatinine umol/L1 (Increase) 
(Female)  
Version 4: 98-161 
Versions 5-7: 134-176 
Version 4: 162-202 
Versions 5-7: 177-221 
Version 4: >202 
Version 5-7: >221 
AST IU/L males (Increase)  
Version 4: 38.1-200 
Version 5: 60.1-200 
Versions 6 and 7: 70.1-200 
201-300 
>300 
AST IU/L females (Increase) 
Version 4: 32.1-200 
Version 5: 60.1-200 
Versions 6 and 7: 70.1-200 
201-300 
>300 
ALT IU/L  males (Increase)  
Version 4: 41.1-200 
Version 5: 60.1-200 
Versions 6 and 7: 70.1-200 
201-300 
>300 
ALT IU/L females (Increase) 
Version 4: 31.1-200 
Version 5: 60.1-200 
Versions 6 and 7: 70.1-200 
201-300 
>300 
 
 
 
 
Urinalysis 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Protein 
1+ 
2+ 
3+ 
or hospitalization or 
dialysis 
1Conversion factor from mg/dL: http://onlinelibrary.wiley.com/doi/10.1002/97808138188825.app3/pdf 
2Trace protein at screening (Visit 00B) is acceptable for inclusion into the study.  
† Values of 60 fall into both Mild and Moderate categories for protocol versions 4-7 and will be classified as Mild. 
 


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5. 
SAMPLE SIZE CONSIDERATIONS 
The primary amebiasis outcome for this study is to compare the proportion of patients/subjects with resolution 
of diarrhea by Day 7 of therapy, in which placebo is expected to have a clearance rate of approximately 15-
20%.  Sample size calculations for both primary endpoints are based on a two-sample, two-sided binomial test 
for proportions, with alpha set to 0.05.  Calculations were performed using the R statistical package (Version 
3.0.2; http://www.r-project.org).  The primary amebiasis endpoint in this aim is to compare the resolution of 
diarrhea between the two study arms to demonstrate that auranofin increases resolution of diarrhea.  For this 
power analysis, we assume the following: a predicted diarrhea resolution rate of at least 80% (based on 
treatment results with current standard therapy with metronidazole [1]) in the auranofin treatment group.   
With these assumptions, assuming 10% attrition (determined from the average of multiple other clinical trials 
in the same Mirpur district), a sample size of 68 with amebiasis (34 per arm; 60 completers in total) achieves 
greater than 95% power to detect a difference between the group proportions of 60%. The high statistical 
power for the primary endpoint is planned to also ensure adequate power for the secondary endpoints.  
For the primary giardiasis endpoint of proportion of subjects with clearance of Giardia by Day 5, a similar 
sample size of 34 per arm will achieve 85% power to detect a 40% difference between treatment arms with a 
predicted clearance rate of 70% (based on response rates to metronidazole or tinidazole, Gardiner, 2001) and a 
30% clearance rates in placebo.  


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6. 
GENERAL STATISTICAL CONSIDERATIONS 
6.1. 
General Principles 
Continuous variables will be summarized using the following descriptive statistics: n (non-missing sample 
size), mean, standard deviation, median, maximum and minimum. The frequency and percentages (based on 
the non-missing sample size) of observed levels will be reported for categorical measures. In general, all data 
will be listed, sorted by disease group, site, treatment and subject, and when appropriate by visit number 
within subject. In general, summary tables will be created separately for the four populations (asymptomatic 
amebiasis, asymptomatic giardiasis, symptomatic amebiasis, and symptomatic giardiasis), and will include 
columns for each treatment in the order (Auranofin, Placebo) and will be annotated with the total population 
size relevant to that table/treatment. 
6.2. 
Timing of Analyses 
There will be no planned interim analyses for efficacy or futility conducted for this study, but the DSMB may 
modify this during ongoing safety monitoring.  
Asymptomatic subjects (34) enrolled under Versions 1-4 of the protocol will have their data reviewed and 
clinical database locked for analysis first and a separate report generated. This may be generated before 
follow-up is completed in the symptomatic study population. 
In the event that recruitment and follow-up for either the Giardiasis arm or the Amebiasis arm is completed 
ahead of the other, the primary and secondary endpoints may be analyzed for that arm using the specified 
methods.  This report will be considered the interim CSR and will contain detailed unblinded information 
about the participants, their immunological response to treatment as well as their side effects and any 
laboratory abnormalities.  The interim CSR will be followed with a release of data corresponding to the 
report. 
6.3. 
Analysis Populations 
The primary analysis population is the modified Intention-to-Treat (mITT) population, described in 6.3.1 
below.  All primary and secondary endpoints will be conducted on both the modified Intention-to-Treat 
population and the per-protocol population, described in Sections 6.3.1 and 6.3.2, respectively.  Tabular 
listings of all subjects excluded from each of these populations are provided, separately for each disease 
group, in Listing 7 and Listing 8.  
6.3.1 Modified Intention-to-Treat (mITT) Population 
In this study, the primary analysis will be performed on the mITT population.  This population was defined in 
the protocol to include study participants who were randomized and dispensed at least one dose of study 
medication.  However, as described in section 6.3.1.1 below, those who failed the eligibility criterion 
regarding testing positive for amebiasis or giardiasis will be excluded.  Excluding subjects who were not 
dispensed study medication has the potential to introduce bias in estimates if the dispensation of medication is 
influenced by treatment assignment.  Although this study is not perfectly blinded, the procedures for 
distributing medication are designed to reduce the possibility of un-blinding the staff dispensing medication 
(see Section 4.4.3).  These procedures ensure that study staff dispensing medication will not be able to 
identify the treatment assignment of subjects prior to dispensing the first dose of treatment.  Therefore, it is 


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unlikely that bias will be introduced by excluding subjects who did not receive at least one dose.  The 
possibility of bias will be assessed by comparing dispensation rates between the treatment and control groups. 
6.3.1.1. 
Exclusions Due to Violation of Eligibility Criteria 
Furthermore, subjects who meet the primary outcome at baseline (and therefore should not have been 
enrolled) will be excluded from the mITT population.  In the asymptomatic population, the outcome is a 
negative microscopy or antigen test at Day 5 or 7, so subjects were required to have both positive microscopy 
result and positive test result for their respective disease in order to enroll.  Those who did not will be 
excluded. 
In the symptomatic population, subjects were to have amebiasis or giardiasis identified by rapid EIA and 
positive antigen detection at enrollment in order to be eligible.  The primary outcome is resolution of diarrhea 
by Day 5 or 7.  Those who either failed the eligibility criterion (i.e., had a negative EIA or negative antigen 
test), or did not have diarrhea at enrollment, will be excluded from the mITT population. 
6.3.2. 
Per Protocol Population 
As a secondary analysis, a per-protocol (PP) population will be analyzed.  The per-protocol population will 
consist of all data from subjects who did not substantially deviate from the protocol during the time leading up 
to (and including) the collection of the data point in question.  Missed or out-of-window stool samples from 
prior visits will not be considered protocol deviations as long as the stool sample for the visit of interest (for a 
particular outcome) is provided within window.  For example, if a stool sample was not provided on Day 3, 
but is provided on Day 5, this stool sample will be included for analysis of endpoints on Day 5.  Subjects who 
completed at least 80% of the treatment regimen will be included in the per-protocol population.  This 
corresponds to at most a single missed dose during the course of treatment, since the amebiasis group has 
seven required doses (and 6/7=86%) and the giardiasis group has five required doses (and 4/5=80%).  If a 
large number of subjects missed a single dose, we will perform sensitivity analysis by also analyzing the 
population of subjects with 100% compliance.  Endpoints that are measured out of window will be excluded 
from this analysis. 
All exclusions described in Section 6.3.1.1 (Exclusions due to violation of eligibility criteria) also apply to the 
PP Population. 
Subjects may also be removed from this population as determined on a per-subject basis by the PI and the 
DMID Scientific Lead prior to unblinding. 
6.3.3. 
Safety Population 
The safety population will consist of all subjects who received any study product (including control).  This 
population will be summarized according to the actual treatment received. 
6.4. 
Covariates and Subgroups 
All analyses will be performed separately for the two disease groups (amebiasis and giardiasis).  Apart from 
disease strata, the protocol does not formally define any subgroup analyses, and the study is not adequately 
powered to perform subgroup analyses. 


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6.5. 
Missing Data 
The primary analysis involves comparison of a binary response between the treatment group and the placebo 
group.  This is true for both asymptomatic and symptomatic populations, whose primary outcomes are 
parasitological response and resolution of diarrhea, respectively.  Within each population, the response is 
measured at Day 7 for the amebiasis group and Day 5 for the giardiasis group.  In the symptomatic 
population, subjects who terminate their treatment regimen and receive metronidazole due to worsening 
symptoms will be imputed as treatment failures for the primary outcome.  That is, they will be imputed as still 
symptomatic by Day 7 in the amebiasis group, and by Day 5 in the giardiasis group.  Secondary outcomes 
measured at Days 3, 5, and 7 will be imputed analogously, with the exception of the quantitative 
trophozoite/cyst load.  
The rest of the text in this section applies to both asymptomatic and symptomatic populations. 
In the primary analysis, no covariates are included, so the analysis involves two variables: the binary predictor 
(treatment/placebo) and the binary response (parasitological response: yes/no).  Some subjects may have 
missing values for the response variable, for example, if they are unable to provide a stool specimen on the 
designated day.  If all subjects are equally likely to have missing response values, then the data are Missing 
Completely At Random (MCAR), in which case these subjects may be dropped from analysis without biasing 
the results.  If, on the other hand, missingness depends on other values which are observed, then the data are 
Missing At Random (MAR).  It is possible that the data in this study will be MAR but not MCAR (which is a 
more stringent condition).  For example, it is plausible that subjects in the auranofin group will be more likely 
to miss a stool specimen than those in the placebo group since constipation is one known possible side effect 
of auranofin.  If this is the case, and if those who are missing response values in the auranofin group also 
experience a different response to treatment than those who remain in the study, then dropping these subjects 
from analysis will bias the estimate of the treatment effect.  An appropriate approach to remove the bias from 
dropping these subjects from analysis is multiple imputation, which is proposed below.  A third possibility for 
the missingness mechanism is that the missingness depends on one or more unobserved variables (and may 
depend on observed variables, as well).  As all relevant symptom variables, as well as demographic variables, 
will be measured, it is unlikely that missingness pattern will depend on an unobserved variable.  For the 
symptomatic population, microscopy results will also be evaluated as a predictor. 
Multiple imputation will be used to handle missing response values for the primary analysis. Multiple 
imputation is a method in which observed covariates are used to predict missing values, and the uncertainty 
arising from this imputation procedure is incorporated into final treatment effect estimates.  Single imputation 
methods, such as imputing treatment failures for all missing results, result in unrealistically small standard 
errors as the imputed counts are less variable than real data. In contrast, multiple imputation methods can 
provide valid standard error estimates. Multiple imputation methods fill in missing values repeatedly, creating 
multiple sets of completed datasets. For each of the datasets, point estimates and standard errors are 
computed. These estimates and standard errors are then combined to obtain the multiple imputation estimate 
and standard error estimate [2]. 
As our response variable is binary, logistic regression will be used for the multiple imputation procedure.  In 
order to determine which covariates to include as predictors of missingness, tests of association between all 
baseline variables and missingness will be performed (t-tests, chi-square tests, or Fisher’s exact tests, as 
appropriate).  If appropriate, t-tests will be performed on transformed data, although sample size is likely 
large enough that transformation will not be required.  Covariates whose p-values are greater than 0.10 will be 
included in the imputation model.  Symptom variables including constipation, diarrhea, anorexia, nausea, 


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abdominal pain, whether or not the subject had any other AE, and whether or not the subject withdrew from 
the study due to an AE will be evaluated for inclusion through the same criterion.  For the symptomatic 
population, microscopy results will be evaluated as well. 
Next, the imputation procedure proceeds as follows: 
1. Imputation step.  In this step, multiple copies of the original data set are generated, and missing values 
of the response variable in each copy are imputed based on a logistic regression model which includes 
baseline variables as predictors as well as study variables measured after baseline.  The imputed data 
sets will vary in their imputed values but will have identical non-missing values.  The generation of 
multiple imputed data sets enables us to quantify the uncertainty arising from the imputation process.  
2. Analysis step.  On each imputed data set, the primary analysis will be performed: computing the 
difference in proportions of parasitological response between treatment and placebo arms, as well as 
the standard error for the difference in proportions.  The odds ratio and respective standard error will 
be computed as well. 
3. Combination step.  The imputed data sets are combined to obtain a single estimate for the difference in 
proportions and standard error, and a single estimate for the odds ratio and standard error. 
The secondary analysis includes a per-protocol analysis including only participants with completely observed 
data.  This analysis requires the data to be MCAR, a more stringent condition than MAR.   
All except one of the secondary outcomes are also binary, so will be analyzed through the same imputation 
method.  The sole numeric outcome is parasite load, measured at multiple time points and analyzed with a 
mixed effects model for longitudinal data.  The mixed effects model, described below, adjusts for data that are 
missing at random (MAR), so imputation is not required in that analysis. 
6.5.1. 
Sensitivity Analyses 
The primary analysis assumes that missing data are missing at random – i.e. the missingness depends only on 
the values of observed variables.  This is an assumption which cannot be proven.  Although we can show that 
missingness is related to values of observed variables (suggesting MAR and proving that MCAR does not 
hold), it is impossible to prove that there is no non-observed variable related to missingness.   
However, we can assess whether MCAR is satisfied – the condition required for our per-protocol analysis.  To 
do so, we will test for associations between all covariates and missingness with t-tests, chi-square tests, and/or 
Fisher’s exact tests, as appropriate.  The results of the logistic regression model predicting missingness based 
on the combination of covariates will also be reported and discussed. 
6.6. 
Interim Analyses and Data Monitoring 
There will be no planned interim analyses for efficacy or futility conducted for this study, but the DSMB may 
modify this during ongoing safety monitoring. The study team will review all adverse events by cumulative 
reports on a monthly basis.  Adverse events will be graded using the protocol defined grading system. The 
DSMB, established by the study team, will monitor this study. 
The DSMB will review the safety data when the study reaches 50% enrollment (i.e. 34 subjects enrolled per 
parasite group).  The DSMB will review safety data after the 68th subject completes visit 8.  


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Asymptomatic subjects (34) enrolled under Version 1-4 of the protocol will have their data reviewed and 
clinical database locked for analysis first and a separate report generated. This may be generated before 
follow-up is completed in the symptomatic study population. 
In the event that recruitment and follow-up for either the Giardiasis arm or the Amebiasis arm is completed 
ahead of the other, the primary and secondary endpoints may be analyzed for that arm using the specified 
methods.  This report will be considered the interim CSR and will contain detailed unblinded information 
about the participants, their immunological response to treatment as well as their side effects and any 
laboratory abnormalities.  The interim CSR will be followed with a release of data corresponding to the 
report.  
6.7. 
Multiple Comparisons/Multiplicity 
For each study population (asymptomatic, symptomatic) and disease group (amebiasis, giardiasis) there is a 
single primary endpoint.  This can be viewed as four parallel studies.  No adjustments for multiple testing are 
planned. 


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7. 
STUDY SUBJECTS 
7.1. 
Disposition of Subjects 
Table 14 will present a summary of the reasons that subjects were screened but not enrolled.  The table will be 
repeated for each study population. 
The disposition of subjects in the study will be tabulated by treatment group, separately for each disease group 
and study population (Table 9 and Table 10).  The table shows the total number of subjects screened, the 
number that enrolled and received at least 1 dose, the number that received all scheduled doses, and the 
number that completed stool sample for the primary endpoint (day 5 for the giardiasis group; day 7 for the 
amebiasis group).  
A flowchart showing the disposition of study subjects, adapted from the Consort Statement (Drummond, 
2001) is included (Figure 3). This figure will present the number of subjects screened, enrolled, lost to follow-
up, and analyzed, by treatment arm.  A separate flowchart will be created for each study population. 
The composition of analysis populations, including reasons for subject exclusion, by treatment arm, is 
presented in Table 11 and Table 12. A separate table is provided for each disease group and study population.  
Listing 1 and Listing 2 will include subjects who discontinued dosing or terminated from study follow-up, and 
the reason will be included.   The listings will be displayed separately for each disease group and study 
population. 
7.2. 
Protocol Deviations 
A summary of subject-specific protocol deviations will be presented by the reason for the deviation, the 
deviation category, and treatment group for all subjects (Table 8), separately for each disease group and study 
population.  All subject-specific protocol deviations and non-subject-specific protocol deviations will be 
included as data listings (Listing 3, Listing 4, Listing 5, and Listing 6, respectively). 


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8. 
EFFICACY EVALUATION 
All efficacy analyses will be performed on the modified Intention-to-Treat (mITT) population, as described in 
section 4.3.1, as a primary analysis.  As a secondary analysis, the analysis will be repeated on the per-protocol 
population, as described in section 4.3.2.  All tests will be two-sided and will use a significance level of 0.05. 
No adjustments will be made for multiple testing.  All results will be reported as point estimates (differences 
in proportions, odds ratios or mean differences across groups, as appropriate) and interval estimates (95% 
confidence intervals). 
8.1. 
Primary Efficacy Analysis 
8.1.1. 
Amebiasis Group 
For the asymptomatic population, the primary amebiasis endpoint is clearance of Entamoeba histolytica by 
day 7, defined as no detection of cysts or trophozoites of Entamoeba histolytica or negative antigen detection 
on day 7.  The differences in parasite clearance rates between the placebo and auranofin arms for Entamoeba 
histolytica will be reported, along with the odds ratio (OR) and their 95% confidence intervals.  The 
confidence interval for the odds ratio will be computed using the assumption of asymptotic normality of the 
log odds ratio, which should hold in this case as sample sizes are 34 in each arm.  The difference in 
proportions is normally distributed for large enough sample sizes unless one or both of the proportions being 
estimated is near the boundary of the parameter space (i.e., zero or one).  We expect normality to hold as we 
expect 15-20% parasitological response in the placebo group and 80% in the treatment group, but if it does 
not, we will use the adjusted Wald estimate as recommended by Agresti and Coull [3]. 
If some subjects are missing their response value, multiple imputation, as described in section 5 above, will be 
used to adjust estimates of treatment effect and standard error.   
The following hypotheses will be tested: 
H0: The probability of parasitological response by Day 7 for subjects receiving auranofin is equal to 
that for those receiving placebo. 
HA: The probability of parasitological response by Day 7 for subjects receiving auranofin differs from 
that for those receiving placebo. 
As a secondary analysis, multivariable logistic regression analysis will be performed to study the association 
between clearance rates and intervention arm, adjusting for baseline demographic and clinical characteristics.  
A series of models will compare each baseline covariate with treatment status and the response.  Baseline 
variables will include demographic variables, baseline vital signs and laboratory values.  Covariates that are 
significantly associated with both treatment group and outcome (p < 0.10) will be selected as predictors in a 
multivariable logistic regression model as these are possible confounding variables.  The logistic regression 
model assumes that continuous covariates are linearly related to the log odds of the response.  To test this 
assumption, we will calculate the empirical log odds of the response repeatedly for different observed 
intervals of the covariate (e.g., calculate the empirical log odds when the covariate is valued 0-5, 5-10, 10-15, 
etc.), and plot the empirical log odds against the covariate interval.  If the linearity assumption holds, this 
graph will show an approximately linear pattern.  Transformations (e.g. the log transformation) for continuous 
covariates showing a nonlinear relationship to the response will be considered. 
Table 18 and Table 20 will present the numbers and proportions (with 95% confidence intervals) of subjects 
with parasitological response by Day 7 for the treatment and placebo arms in the amebiasis strata for the 


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asymptomatic and symptomatic populations, respectively.  These tables also present the difference in 
proportions between treatment and placebo groups and odds ratios, with their 95% confidence intervals.  
Separate versions of the tables will be provided for the modified-Intention-to-Treat population and the Per-
Protocol population. 
In the symptomatic population, the primary outcome is resolution of diarrhea by Day 7.  This variable will be 
analyzed with the same approach above described for the analysis of parasitological response in the 
asymptomatic population.  The following hypotheses will be tested: 
H0: The probability of resolution of diarrhea by Day 7 for subjects receiving auranofin is equal to that 
for those receiving placebo. 
HA: The probability of resolution of diarrhea by Day 7 for subjects receiving auranofin differs from 
that for those receiving placebo. 
8.1.2. 
Giardiasis Group 
The primary endpoint for the giardiasis group is clearance of Giardiasis by day 5, defined as no detections of 
cysts or trophozoites of Giardia or negative antigen detection on day 5.  This endpoint is analogous to the 
amebiasis endpoint except for the time point, and analogous methods will be applied. Table 19 and Table 21 
will present the numbers and proportions (with 95% confidence intervals) of subjects with parasitological 
response by Day 7 for the treatment and placebo arms in the giardiasis strata for the asymptomatic and 
symptomatic populations, respective.  These tables also present the difference in proportions between 
treatment and placebo groups and odds ratios, with their 95% confidence intervals.  Separate versions of the 
tables will be provided for the modified-Intention-to-Treat population and the Per-Protocol population.   
In the symptomatic population, the primary outcome is resolution of diarrhea by Day 5.  This variable will be 
analyzed with the same approach above described for the analysis of parasitological response.   
8.2. 
Secondary Efficacy Analyses 
8.2.1. 
Amebiasis Group 
The secondary outcome measures for the asymptomatic amebiasis group are: 
1. Parasitological response (no detection of cysts or trophozoites on microscopic exam or negative 
antigen detection) by Day 3 and 5 
2. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3,5 and 7 
3. Proportion of subjects with negative stool antigen test by Days 3, 5, 7, and 14 
4. Proportion of subjects with sustained cure (no detection of cysts or trophozoites by microscopic exam 
or negative antigen detection) at 14 and 28 days 
5. Proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 14 
and 28 days by genotyping the initial vs. subsequent strain. 
Secondary outcome (1) corresponds to the primary outcome, except that the time points of measurement 
differ.  Therefore, methods perfectly analogous to those used for the primary outcome will be applied to the 
first secondary outcome.  Secondary outcomes (3) – (5) above are also dichotomous, so will be analyzed with 
methods analogous to those described for the primary outcome (e.g. odds ratios with 95% confidence 
intervals, and/or logistic regression with confounders included as predictors).   


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Secondary aim (2) includes the sole numeric outcome (parasite load), and this is measured at multiple time 
points.  For secondary aim (2), a mixed effects model will be used to compare the rate of change in parasite 
loads between the two study arms.  A mixed effects model allows us to incorporate information for multiple 
time points into a single model while explicitly adjusting for the correlation (due to repeated measures on the 
same subjects) that naturally arises from this data structure. [4] Below the mixed effects model for the per-
protocol analysis is described.  This is followed by a description of how the model will be adjusted in the m-
ITT population analysis in order to account for missing data. 
Parasite load measured on study days 1, 3, 5, and 7 will be included in the mixed effects model.  The mean 
parasite load with 95% confidence intervals will be displayed graphically for each time point for each 
treatment group, as shown in the example figure below: 
 
If a linear relationship between parasite load and time is observed for both groups, then a model that makes 
use of this simplifying assumption will be used.  Letting Y denote parasite load, t denote the time point (study 
day), and i be a subject index, and letting treatment=1 for those in the treatment group and treatment=0 for 
those in the placebo group, we will model parasite load as: 
𝑌𝑌𝑖𝑖𝑖𝑖= 𝛽𝛽0 + 𝛽𝛽1𝑡𝑡+ 𝜇𝜇𝑖𝑖+ 𝛼𝛼𝑖𝑖𝑡𝑡+ 𝛽𝛽2𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡+ 𝛽𝛽3𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡∙𝑡𝑡+  𝜖𝜖, 
Note that the above graph does not depict a linear decline in parasite load over time.  If the relationship is 
nonlinear, as depicted in the figure above, then transformation (e.g. log transformation) may be applied, or we 
will fit the following model, which relaxes the linearity assumption.  Let 1[t=k] denote an indicator variable 
taking the value one when t=k and zero otherwise. Let ui denote the random intercept for subject i, and let αi 
denote the random slope for subject i, and assume these random effects are normally distributed with mean 
zero.  Then without assuming linearity, the parasite load for subject i at time t can be modeled as:  
𝑌𝑌𝑖𝑖𝑖𝑖= 𝛽𝛽0 + 𝜇𝜇𝑖𝑖+ 𝛼𝛼𝑖𝑖𝑡𝑡+ 𝛽𝛽1 ∙𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡+ 𝛽𝛽2 1[t=3] + 𝛽𝛽3 1[t=3] ∙𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡+ 𝛽𝛽41[t=5] + 𝛽𝛽5 1[t=5]
∙𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡+ 𝛽𝛽41[t=7] + 𝛽𝛽5 1[t=7] ∙𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡+  𝜖𝜖, 
The error terms, 𝜖𝜖,  are assumed to be independent and normally distributed with constant variance and mean 
zero. If residual plots indicate that variance is not constant and/or the errors are not normally distributed, 


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transformations (e.g., the log transformation of the response) will be considered.  Choice of covariance 
structures for the random effects (e.g., compound symmetric and autoregressive), will be explored. 
In this model, the expected parasite load for the placebo group on Day 1 is 𝛽𝛽0, and on Day 3 (t=3) is 𝛽𝛽0 + 𝛽𝛽2.  
Therefore, the expected decrease from time Day 1 to Day 3 is 𝛽𝛽2 for the placebo group. The expected parasite 
load for the treatment group on Day 1 is 𝛽𝛽0 + 𝛽𝛽1, and on Day 3, it is 𝛽𝛽0 + 𝛽𝛽1 + 𝛽𝛽2 + 𝛽𝛽3.  Therefore, the 
expected decrease for the treatment group from Day 1 to Day 3 is 𝛽𝛽2 + 𝛽𝛽3. Thus, the difference in rate of 
decrease of parasite load from Day 1 to Day 3 between treatment and control groups is, on average, 𝛽𝛽3.  The 
parameters 𝛽𝛽5 and 𝛽𝛽7 are interpreted analogously. 
The per-protocol analysis will be performed as described above.  The per-protocol population, by definition, 
has no missing data.  For the m-ITT analysis, if there is missing outcome data, the model will be altered to 
adjust for missing response values, which we assume, as previously described, to be missing at random.  
Covariates that are associated with missingness (p>0.10 via t-test, Fisher’s exact test, or chi-square test, as 
appropriate) will also be included as explanatory variables in the model.  This model naturally adjusts for 
values that are missing at random, so multiple imputation is not required. 
The secondary outcomes will be summarized in Table 22 through Table 34.  Table 22 and Table 24 will 
present clearance rates for the amebiasis group with 95% confidence intervals on Days 3, 5, and 7 for the 
treatment and placebo groups, as well as the differences in clearance rates and 95% confidence intervals for 
the differences.  Table 23 and Table 25 are analogous tables for the giardiasis group. Table 26 will present the 
coefficient estimates for the mixed effects model for the amebiasis group.  Table 27 is the analogous table for 
the giardiasis group. Table 28 will present the proportion of subjects in the amebiasis group with negative 
stool antigen test on Days 1, 3, 5, 7, and 14.  This outcome is not analyzed for the giardiasis group. Table 29 
will present the proportion of subjects with sustained cure for the amebiasis group on days 7, 14, and 28, for 
each treatment arm, with 95% confidence intervals, as well as the difference in proportions between treatment 
arms and 95% confidence interval for the difference.  Table 30 will present analogous summaries for the 
giardiasis group (Day 5 instead of Day 7).  Table 31 will present the proportion of subjects in the amebiasis 
group with relapse by Day 14, the proportion with reinfection by Day 14, and the proportion with either 
relapse or reinfection by Day 14, separately for each treatment arm, with 95% confidence intervals.  Table 32 
is the analogous table for the giardiasis group.  Differences in proportions between treatment arms and 95% 
confidence intervals for the differences will also be included.  Table 33 will present a perfectly analogous 
summary for Day 28 rather than Day 14, for the amebiasis group.  Table 34 is the analogous table for the 
giardiasis group. Listing 15 and Listing 16 provide a listing for all efficacy endpoints for the amebiasis 
groups; Listing 17 and Listing 18 are the analogous listings for the giardiasis groups. 
The secondary outcomes for the symptomatic population are identical to those for the asymptomatic 
population with the following exceptions: 
• Parasitological response is defined differently, and parasitological response on Day 5 (giardiasis) 
and Day 7 (amebiasis) were changed from primary to secondary. 
• Time to resolution of diarrhea was added as an endpoint. 
The outcomes that are identical to the asymptomatic population will be analyzed identically.  Parasitological 
response on Day 5 (giardiasis) and Day 7 (amebiasis) will be analyzed by the methods planned for 
parasitological response on the other study days, described above.  Kaplan-Meier curves and log rank tests 
will be used to compare time to diarrhea resolution between the treatment and control arm for each disease 
group (Figure 4). 


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8.2.2. 
Giardiasis Group 
The secondary outcomes for the asymptomatic giardiasis group are: 
6. Parasitological response (no detection of cysts or trophozoites on microscopic exam or negative 
antigen detection) by Day 3 
7. Rate of decrease of trophozoite/cyst load by qPCR in stools by Days 3,5 
8. Proportion of subjects with sustained cure (no detection of cysts or trophozoites by microscopic exam 
or negative antigen detection) at 14 and 28 days 
9. Proportion of subjects with relapse (same strain) or re-infection (new strain) with positive stools at 14 
and 28 days by genotyping the initial vs. subsequent strain 
These outcomes are analogous to the asymptomatic amebiasis outcomes and will be analyzed using perfectly 
analogous methods.  
Similarly, the secondary outcomes for the symptomatic giardiasis population will be analyzed analogously to 
those for the symptomatic amebiasis population 


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9. 
SAFETY EVALUATION 
9.1. 
Demographic and Other Baseline Characteristics 
Summaries of age, sex, ethnicity, and race will be presented by treatment group and overall (Table 15 and 
Table 16), for each disease group and study population.  Ethnicity is categorized as “Hispanic or Latino”, or 
“Not Hispanic or Latino”.  In accordance with NIH reporting policy, subjects may self-designate as belonging 
to more than one race or may refuse to identify a race, the latter reflected in the CRF as “No” to each racial 
option. 
Individual subject listings will be presented for all demographics (Listing 9 and  Listing 10); pre-existing 
medical conditions (Listing 11 and Listing 12); vital signs and oral temperature (Listing 31 and Listing 32); 
and concomitant medications (Listing 35 and Listing 36). These listings will be presented separately for each 
disease group and study population. 
9.1.1. 
Prior and Concurrent Medical Conditions 
All current illnesses and past pre-existing medical conditions will be MedDRA coded using MedDRA 
dictionary version 18.1 or higher.  Summaries of subjects’ pre-existing medical conditions will be presented 
by treatment group (Table 17), disease group, and study population.  Individual subject listings will be 
presented for all medical conditions (Table 11 and Table 12).  Listings are presented separately for each 
disease group and study population. 
9.1.2. 
Prior and Concomitant Medications  
Summaries of medications that were started prior to dosing and continuing at the time of dosing will be 
presented by WHO Drug classification and treatment group, separately for each disease group and study 
population (Table 79 and Table 80).  Individual subject listings will be presented for all concomitant 
medications (Listing 35 and  Listing 36). 
9.2. 
Measurements of Treatment Compliance 
In the amebiasis group, all subjects were to receive a total of 7 doses of study product (once a day for 7 days). 
In the giardiasis group, all subjects were to receive a total of 5 doses of study product (once a day for 5 days).   
The number of doses of study product administered to subjects will be presented by treatment group as part of 
the subject disposition tables, separately for each disease group and study population (Table 9 and Table 10).  
Listing 13 and Listing 14 list the doses missed and treatment assignment for each subject. 
Table 13 summarizes dosing dates by treatment arm, disease group, and study population. 
9.3. 
Adverse Events 
9.3.1. 
Solicited Events and Symptoms 
Adverse event data is collected at each visit using specific questions and/or targeted physical examination. 
Based on the adverse reactions that have been observed with the use of auranofin in previous studies, the 
following solicited events are tracked: loose stools or diarrhea, abdominal pain, nausea with or without 
vomiting, constipation, anorexia, flatulence, dyspepsia, dysgeusia, pruritus, hair loss, urticaria, stomatitis, 
conjunctivitis, glossitis, hematuria, and rash.  Hematologic, renal, or hepatic laboratory abnormalities are 


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picked up on testing at day 7. Adverse event collection continues until Day 28 (following three half-lives of 
the drug).  As the drug is administered orally, no local event reactivity is recorded. 
Table 35 and Table 36 display the number, percentage, and 95% confidence interval for the percentage of 
subjects experiencing each solicited adverse event, by treatment group, separately for the amebiasis group and 
the giardiasis group.  Solicited adverse events are also displayed graphically.  Figure 5 displays the maximum 
severity of solicited symptoms per subject by day post treatment, for each treatment group and for both 
treatment groups combined, separately for each disease group and study population.   
Solicited systemic events by subject will be presented in Listing 19, Listing 20, Listing 21, and Listing 22. 
9.3.2. 
Unsolicited Adverse Events 
Unsolicited adverse events will be coded to a Medical Dictionary for Regulatory Activities Terminology 
(MedDRA) version 18.1 or later. Adverse Events will be summarized using the number and percentage of 
subjects who experienced each event overall for the safety population.  When calculating the incidence of 
adverse events (i.e., on a per subject basis), each subject will only be counted once for each PT and SOC and 
any repetitions of adverse events within a subject for a PT or SOC will be ignored; the denominator will be 
the total number of subjects in the safety population. All adverse events reported will be included in the 
summaries and analyses.   
Table 37 through Table 40 summarize the number and percentage of subjects experiencing unsolicited 
adverse events by MedDRA System Organ Class and Preferred Term, severity, and relationship, by treatment, 
disease group, and study population.   
A listing of serious adverse events, sorted by study population, disease group, and treatment group, is 
presented in Table 41.  Listings of non-serious, unsolicited, moderate or severe adverse events, separately for 
each disease group and study population, are presented in Table 42 and Table 43.  
Unsolicited adverse events are also displayed graphically.  Figure 6 and Figure 7 display the frequency of 
unsolicited adverse events by MedDRA System Organ Class by treatment group, separately for each disease 
group and study population.  Figure 8 and Figure 9 display the incidence of adverse events by MedDRA 
System Organ Class by treatment group, separately for each disease group and study population.  Figure 10 
and Figure 11 summarize the frequency of unsolicited adverse events by severity for each treatment group and 
overall, separately for each disease group and study population.  Figure 12 and Figure 13 display the 
incidence of unsolicited adverse events by severity for each treatment group and overall, separately for each 
disease group and study population.  Figure 14 and Figure 15 display the frequency of unsolicited adverse 
events by MedDRA System Organ Class and relationship to treatment, by treatment group, separately for 
each disease group and study population.  Figure 16 and Figure 17 display the incidence of unsolicited 
adverse events by MedDRA System Organ Class and relationship to treatment, by treatment group, separately 
for each disease group and study population.  Figure 18 and Figure 19 display the frequency of unsolicited 
adverse events by relationship to treatment, by treatment group, separately for each disease group and study 
population.  Figure 20 and Figure 21 display the incidence of unsolicited adverse events by relationship to 
treatment, by treatment group, separately for each disease group and study population. 
Unsolicited adverse events by subject will be presented in Listing 23 and Listing 24, separately for each 
disease group and study population. 


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9.4. 
Deaths, Serious Adverse Events and other Significant Adverse Events 
Section 14.3.3 will include narratives of deaths, and other serious and significant adverse events. 
9.5. 
Pregnancies 
Auranofin is a Category C drug whose use is not recommended in pregnant women because of studies in 
pregnant rabbits and rats with decreased maternal and fetal weight and litter size with 4-50X the human daily 
dose. This trial excludes pregnant women based on a pregnancy test during screening and requires two forms 
of contraception for the length of the trial (5-7 days) and for three half-lives of the drug (total 4 months).  For 
any subjects in the Safety population who became pregnant during the study, study medication was stopped.  
Every attempt will be made to follow these subjects to completion of pregnancy to document the outcome, 
including information regarding any complications with pregnancy and/or delivery.   A listing of pregnancies 
and outcomes will be presented (Listing 37 through Listing 41). 
9.6. 
Clinical Laboratory Evaluations 
Laboratory parameters include white blood cell count, hemoglobin, hematocrit, platelets, BUN, Creatinine, 
AST, ALT, and urine protein, all measured during screening and on Study Day 7.  Units and severity grading 
for the labs are summarized in Table 7. 
The distribution of laboratory results by maximum severity, study day and treatment group will be presented, 
separately for each disease group and study population, in Table 46 through Table 50. Abnormal laboratory 
results related to study treatment will be summarized by maximum severity, study day, and treatment group in 
Table 51 through Table 58, separately by disease group and study population.  Descriptive statistics including 
mean, standard deviation, median, minimum and maximum values by study day, for each laboratory 
parameter, will be summarized separately for each disease group and study population in Table 59 through 
Table 66.  Note that in general, summaries include only scheduled labs: unscheduled follow-up labs will be 
excluded from summaries but included in the listings.  These include summaries at each time point as well as 
summaries in the change in observed values from baseline to Day 7.   Listings of abnormal laboratory events 
(one listing for each disease group and study population) will be included in Table 44 and Table 45.  Time 
trends of laboratory values will be graphically displayed in Figure 22 and Figure 23.  Listings for all 
laboratory values will be provided, separately for each disease group and study population, in Listing 25 
through Listing 30. 
9.7. 
Vital Signs and Physical Evaluations 
Vital signs include height and weight, temperature, systolic and diastolic blood pressure, and pulse.  These 
were assessed during screening and on every scheduled visit.  Targeted physical exams were conducted on the 
same days and looked for abdominal pain, hair loss, urticaria, rashes, stomatitis, conjunctivitis, and glossitis.  
Refer to Table 6 for vital sign grading criteria. 
Vital signs will be tabulated by visit, severity, and treatment group, separately for each disease group and 
study population (Table 67 through Table 78). Vital sign listings will be provided for subjects in the Safety 
population (Listing 31 and Listing 32).  Physical exam findings will be listed in Listing 33 and Listing 34. 


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9.8. 
Concomitant Medications 
Concomitant medications will be coded to the Anatomical Therapeutic Classification using the WHO Drug 
Dictionary.  The use of prior and concomitant medications taken during the study will be recorded on the 
CRFs. A by-subject listing of concomitant medication use will be presented (Listing 35 and Listing 36).  The 
use of concomitant medications during the study will be summarized by WHO drug classification and 
treatment group, separately for each disease group and study population, for the Safety population (Table 79 
and Table 80). 


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10. 
PHARMACOKINETICS 
Not applicable. 


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11. 
IMMUNOGENICITY 
Not applicable. 


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12. 
OTHER ANALYSES 
Not applicable. 


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13. 
REPORTING CONVENTIONS 
P-values ≥0.001 will be reported to 3 decimal places; p-values less than 0.001 will be reported as “<0.001”. 
The mean, standard deviation, and any other statistics other than quantiles, will be reported to one decimal 
place greater than the original data. Quantiles, such as median, or minimum and maximum will use the same 
number of decimal places as the original data. Proportions will be presented as two decimal places; values 
<0.01 will be presented as “<0.01”. Percentages will be reported to the nearest whole number; values <1% 
will be presented as “<1”. Estimated parameters, not on the same scale as raw observations (e.g. regression 
coefficients) will be reported to 3 significant figures. 


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14. 
TECHNICAL DETAILS 
SAS version 9.4 or above and/or R version 3.2.2 or above will be used to generate all tables, figures and 
listings and perform statistical analyses. 


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15. 
SUMMARY OF CHANGES IN THE CONDUCT OF THE STUDY OR 
PLANNED ANALYSES 
Version 2.0 of the Statistical Analysis Plan incorporates the following updates: 
• Updated description of design to add symptomatic population and new site. These were added in 
protocol updates since the first version of the SAP. 
• Endpoints and analysis plans for symptomatic population were added. 
• Updated definition of mITT and PP analysis populations to handle unanticipated enrollment of 
participants who did not test positive for the disease. 
• Updated eligibility criteria and grading ranges based on protocol updates. 
• Updated covariance structure options for mixed effects model to facilitate comparison between 
mITT and PP populations. 
• Added possibility of transformation (e.g. log transformation) of response for mixed effects model 
to attain better fit. 
• Removed tables summarizing solicited events by symptom, treatment group, day, and severity and 
unsolicited adverse events by MedDRA SOC and PT, treatment group, and day, because this level 
of granularity is not needed for the small numbers of events that have been observed. 
• Replaced change from baseline plots for lab parameters with time trend plots displaying grading 
ranges. 
• Removed analysis plans relating to Versions 1.0-3.0 of the protocol since the earliest subjects 
enrolled under Version 4.0.  


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16. 
REFERENCES 
1. Pherson, P.O. and Bengtsson, E. 1984. A long-term follow up study of amebiasis treated with 
metronidazole. Scand. J. Infect. Dis. 16:195-8.  
2. Fleiss, Joseph L., Bruce Levin, and Myunghee Cho Paik, Statistical Methods for Rates and 
Proportions.  John Wiley and Sons, 2013. 
3. Agresti, Alan, and Brent A. Coull.  Approximate is better than “exact” for interval estimation of 
binomial proportions.  The American Statistician 52.2 (1998): 119-1 
4. Diggle, Peter, Patrick Heagerty, Kung-Yee Liang, and Scott Zeger. Analysis of longitudinal data. OUP 
Oxford, 2013. 
5. Mallinckrod, Craig H., et al.  Recommendations for the primary analysis of continuous endpoints in 
longitudinal clinical trials. Drug Information Journal 42.4 (2008): 303-319. 


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17. 
LISTING OF TABLES, FIGURES, AND LISTINGS  
Table, figure, and listing shells are presented in Appendices 1, 2, and 3. 
 


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APPENDICES 


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APPENDIX 1. TABLE MOCK-UPS 
 


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LIST OF TABLES 
Table 1: 
Summary of Study Population, Study Design, and Primary Outcome by 
Protocol Version .........................................................................................................10 
Table 2: 
Secondary Outcomes for the Asymptomatic Study Population .................................10 
Table 3: 
Secondary Outcomes for the Symptomatic Study Population ....................................11 
Table 4: 
 Efficacy Variables for the Asymptomatic Study Population .....................................25 
Table 5: 
Efficacy Variables and Time Points for the Symptomatic Study Population .............26 
Table 6: 
 Vital Signs Grading Scale ..........................................................................................28 
Table 7: 
Laboratory Adverse Event Grading Scale ..................................................................29 
Table 8: 
Distribution of Protocol Deviations by Category, Type, and Treatment 
Group, Asymptomatic Population ..............................................................................61 
Table 9: 
Subject Disposition by Treatment Group, Amebiasis Group, Asymptomatic 
Population ...................................................................................................................64 
Table 10: Subject Disposition by Treatment Group, Giardiasis Group, Asymptomatic 
Population ...................................................................................................................65 
Table 11: Analysis Populations by Treatment Group, Amebiasis Group, Asymptomatic 
Population ...................................................................................................................66 
Table 12: Analysis Populations by Treatment Group, Giardiasis Group ...................................66 
Table 13: Dates of First Treatment by Disease Group and Treatment Group, 
Asymptomatic Population ..........................................................................................67 
Table 14: Ineligibility Summary of Screen Failures, Asymptomatic Population .......................68 
Table 15: Distribution of Sex, Ethnicity, and Race by Treatment Group, Asymptomatic 
Population ...................................................................................................................71 
Table 16: Age Statistics (in Years) by Disease Group and Treatment Group, 
Asymptomatic Population ..........................................................................................72 
Table 17: Distribution of Subjects with Pre-Existing Medical Conditions by MedDRA 
System Organ Class, Disease Group, and Treatment Group, Asymptomatic 
Population ...................................................................................................................73 
Table 18: Comparison of Clearance Rates between Auranofin Group and Placebo 
Group, Day 7 (primary endpoint), Amebiasis Group, Asymptomatic 
Population ...................................................................................................................74 
Table 19: Comparison of Clearance Rates between Auranofin Group and Placebo 
Group, Day 5 (primary endpoint), Giardiasis Group, Asymptomatic 
Population ...................................................................................................................74 
Table 20: Comparison of Resolution of Diarrhea between Auranofin Group and 
Placebo Group, Day 7 (primary endpoint), Amebiasis Group, Symptomatic 
Population ...................................................................................................................75 


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Table 21: Comparison of Resolution of Diarrhea between Auranofin Group and 
Placebo Group, Day 5 (primary endpoint), Giardiasis Group, Symptomatic 
Population ...................................................................................................................75 
Table 22: Comparison of Clearance Rates between Auranofin Group and Placebo 
Group, Asymptomatic Amebiasis Group ...................................................................76 
Table 23: Comparison of Clearance Rates between Auranofin Group and Placebo 
Group, Asymptomatic Giardiasis Group ....................................................................76 
Table 24: Comparison of Clearance Rates between Auranofin Group and Placebo 
Group, Symptomatic Amebiasis Group ......................................................................77 
Table 25: Comparison of Clearance Rates between Auranofin Group and Placebo 
Group, Symptomatic Giardiasis Group ......................................................................77 
Table 26: Mixed Effects Model Coefficient Estimates, Asymptomatic Amebiasis 
Group ..........................................................................................................................78 
Table 27: Mixed Effects Model Coefficient Estimates, Asymptomatic Giardiasis 
Group ..........................................................................................................................79 
Table 28: Comparison of Proportion of Subjects with Negative Stool Antigen Test 
between Auranofin Group and Placebo Group, Asymptomatic Amebiasis 
Group ..........................................................................................................................80 
Table 29: Comparison of Proportion of Subjects with Sustained Cure between 
Auranofin Group and Placebo Group, Asymptomatic Amebiasis Group ..................81 
Table 30: Comparison of Proportion of Subjects with Sustained Cure between 
Auranofin Group and Placebo Group, Asymptomatic Giardiasis Group ...................82 
Table 31: Comparison of Proportion of Subjects with Relapse (Same Strain), and/or 
Reinfection (Different Strain) by Day 14, - Asymptomatic Amebiasis Group ..........83 
Table 32: Comparison of Proportion of Subjects with Relapse (Same Strain), and/or 
Reinfection (Different Strain) by Day 14, - Asymptomatic Giardiasis Group ...........83 
Table 33: Comparison of Proportion of Subjects with Relapse (Same Strain), and/or 
Reinfection (Different Strain) by Day 28, -Asymptomatic Amebiasis Group ...........84 
Table 34: Comparison of Proportion of Subjects with Relapse (Same Strain), and/or 
Reinfection (Different Strain) by Day 28, -Asymptomatic Giardiasis Group ............84 
Table 35: Number and Percentage of Subjects Experiencing Solicited Events with 95% 
Confidence Intervals by Symptom and Treatment Group, - Asymptomatic 
Amebiasis Group ........................................................................................................85 
Table 36: Number and Percentage of Subjects Experiencing Solicited Events with 95% 
Confidence Intervals by Symptom and Treatment Group, Asymptomatic 
Giardiasis Group .........................................................................................................87 
Table 37: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA System Organ Class and Preferred Term, Maximum 


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Severity, Relationship, Asymptomatic Amebiasis Group, Auranofin, 6 mg 
(n=x) ...........................................................................................................................89 
Table 38: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA System Organ Class and Preferred Term, Maximum 
Severity, Relationship, Asymptomatic Amebiasis Group, Placebo (n=x)..................90 
Table 39: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA System Organ Class and Preferred Term, Maximum 
Severity, Relationship, Asymptomatic Giardiasis Group, Auranofin, 6 mg 
(n=x) ...........................................................................................................................91 
Table 40: Number and Percentage of Subjects Experiencing Unsolicited Adverse 
Events by MedDRA System Organ Class and Preferred Term, Maximum 
Severity, Relationship, Asymptomatic Giardiasis Group, Placebo (n=x) ..................92 
Table 41: Listing of Serious Adverse Events .............................................................................93 
Table 42: Listing of Non-Serious, Unsolicited, Moderate or Severe Adverse Events, 
Asymptomatic Amebiasis Group ................................................................................94 
Table 43: Listing of Non-Serious, Unsolicited, Moderate or Severe Adverse Events, 
Asymptomatic Giardiasis Group ................................................................................95 
Table 44: Listing of Abnormal Laboratory Results, Asymptomatic Amebiasis Group .............97 
Table 45: Listing of Abnormal Laboratory Results, Asymptomatic Giardiasis Group ..............97 
Table 46: Laboratory Results by Parameter, Maximum Severity, Study Day, and 
Treatment Group, Asymptomatic Amebiasis Group -   Any Laboratory 
Parameter ....................................................................................................................98 
Table 47: Laboratory Results by Parameter, Maximum Severity, Study Day, and 
Treatment Group, Asymptomatic Amebiasis Group - White Blood Cell 
Count...........................................................................................................................98 
Table 48: Laboratory Results by Parameter, Maximum Severity, Study Day, and 
Treatment Group, Asymptomatic Amebiasis Group - Hemoglobin ...........................99 
Table 49: Laboratory Results by Parameter, Maximum Severity, Study Day, and 
Treatment Group, Asymptomatic Amebiasis Group - Hematocrit .............................99 
Table 50: Laboratory Results by Parameter, Maximum Severity, Study Day, and 
Treatment Group, Asymptomatic Amebiasis Group - Platelets ...............................100 
Table 51: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and Treatment Group, Asymptomatic 
Amebiasis Group -White Blood Cell Count .............................................................101 
Table 52: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and  Treatment Group, Asymptomatic 
Amebiasis Group - Hemoglobin ...............................................................................101 


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Table 53: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and  Treatment Group, Asymptomatic 
Amebiasis Group - Hematocrit .................................................................................102 
Table 54: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and  Treatment Group, Asymptomatic 
Amebiasis Group - Platelets .....................................................................................102 
Table 55: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and Treatment Group, Asymptomatic 
Giardiasis Group -White Blood Cell Count..............................................................103 
Table 56: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and Treatment Group, Asymptomatic 
Giardiasis Group - Hemoglobin................................................................................103 
Table 57: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and Treatment Group, Asymptomatic 
Giardiasis Group - Hematocrit ..................................................................................104 
Table 58: Abnormal Laboratory Results Related to Study Treatment by Parameter, 
Maximum Severity, Study Day, and Treatment Group, Asymptomatic 
Giardiasis Group - Platelets ......................................................................................104 
Table 59: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Amebiasis Group -  White Blood Cells ...............................105 
Table 60: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Amebiasis Group - Hemoglobin ..........................................105 
Table 61: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Amebiasis Group -Hematocrit .............................................106 
Table 62: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Amebiasis Group - Platelets.................................................106 
Table 63: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Giardiasis Group -  White Blood Cells ................................107 
Table 64: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Giardiasis Group - Hemoglobin ...........................................107 
Table 65: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Giardiasis Group - Hematocrit .............................................108 
Table 66: Laboratory Summary Statistics by Parameter, Study Day, and Treatment 
Group, Asymptomatic Giardiasis Group - Platelets .................................................108 
Table 67: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Amebiasis Group - Any Assessment ............................................................109 
Table 68: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Amebiasis Group - Fever ..............................................................................110 


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Table 69: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Amebiasis Group - Systolic Blood Pressure 
(Hypertension/Hypotension) .....................................................................................111 
Table 70: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Amebiasis Group - Diastolic Blood Pressure - Hypertension ......................112 
Table 71: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Amebiasis Group -Bradycardia ....................................................................113 
Table 72: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Amebiasis Group - Tachycardia ...................................................................114 
Table 73: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Giardiasis Group - Any Assessment.............................................................115 
Table 74: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Giardiasis Group - Fever ..............................................................................116 
Table 75: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Giardiasis Group - Systolic Blood Pressure - 
(Hypertension/Hypotension) .....................................................................................117 
Table 76: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Giardiasis Group - Diastolic Blood Pressure - Hypertension .......................118 
Table 77: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Giardiasis Group - Bradycardia ....................................................................119 
Table 78: Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment 
Group, Giardiasis Group - Tachycardia ....................................................................120 
Table 79: Number and Percentage of Subjects with Prior and Concurrent Medications 
by WHO Drug Classification and Treatment Group, Asymptomatic 
Amebiasis Group ......................................................................................................121 
Table 80: Number and Percentage of Subjects with Prior and Concurrent Medications 
by WHO Drug Classification and Treatment Group, Asymptomatic 
Giardiasis Group .......................................................................................................122 
 


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10.2 
Protocol Deviations 
Table 8: 
Distribution of Protocol Deviations by Category, Type, and Treatment Group, Asymptomatic Population 
This table will be repeated for the symptomatic population. 
Category 
Deviation Type 
Amebiasis Group, 
Auranofin, 
6 mg 
(N=34) 
Amebiasis Group, 
Placebo 
(N=34) 
Giardiasis Group, 
Auranofin, 
6 mg 
(N=34) 
Giardiasis Group, 
Placebo 
(N=34) 
All Subjects 
(N=136) 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
Eligibility/enrollment 
Any type 
x 
x 
x 
x 
x 
x 
x 
x 
x 
x 
 
Did not meet inclusion criterion 
 
 
 
 
 
 
 
 
 
 
 
Met exclusion criterion 
 
 
 
 
 
 
 
 
 
 
 
ICF not signed prior to study procedures 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
Treatment administration 
schedule 
Any type 
 
 
 
 
 
 
 
 
 
 
 
Out of window visit 
 
 
 
 
 
 
 
 
 
 
 
Missed visit/visit not conducted 
 
 
 
 
 
 
 
 
 
 
 
Missed treatment administration 
 
 
 
 
 
 
 
 
 
 
 
Delayed treatment administration 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
Follow-up visit schedule 
Any type 
 
 
 
 
 
 
 
 
 
 
 
Out of window visit 
 
 
 
 
 
 
 
 
 
 
 
Missed visit/visit not conducted 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
Protocol procedure/assessment 
Any type 
 
 
 
 
 
 
 
 
 
 


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Category 
Deviation Type 
Amebiasis Group, 
Auranofin, 
6 mg 
(N=34) 
Amebiasis Group, 
Placebo 
(N=34) 
Giardiasis Group, 
Auranofin, 
6 mg 
(N=34) 
Giardiasis Group, 
Placebo 
(N=34) 
All Subjects 
(N=136) 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
 
Incorrect version of ICF signed 
 
 
 
 
 
 
 
 
 
 
 
Blood not collected 
 
 
 
 
 
 
 
 
 
 
 
Urine not collected 
 
 
 
 
 
 
 
 
 
 
 
Stool not collected 
 
 
 
 
 
 
 
 
 
 
 
Other specimen not collected 
 
 
 
 
 
 
 
 
 
 
 
Too few aliquots obtained 
 
 
 
 
 
 
 
 
 
 
 
Specimen result not obtained 
 
 
 
 
 
 
 
 
 
 
 
Required procedure not conducted 
 
 
 
 
 
 
 
 
 
 
 
Required procedure done incorrectly 
 
 
 
 
 
 
 
 
 
 
 
Study product temperature excursion 
 
 
 
 
 
 
 
 
 
 
 
Specimen temperature excursion 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
Treatment administration 
Any type 
 
 
 
 
 
 
 
 
 
 
 
Required procedure done incorrectly 
 
 
 
 
 
 
 
 
 
 
 
Study product temperature excursion 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
Blinding policy/procedure 
Any type 
 
 
 
 
 
 
 
 
 
 


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Table 8: 
Distribution of Protocol Deviations by Category, Type, and Treatment Group, Asymptomatic Population (continued) 
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Category 
Deviation Type 
Amebiasis Group, 
Auranofin, 
6 mg 
(N=34) 
Amebiasis Group, 
Placebo 
(N=34) 
Giardiasis Group, 
Auranofin, 
6 mg 
(N=34) 
Giardiasis Group, 
Placebo 
(N=34) 
All Subjects 
(N=136) 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
# of 
Subj. 
# of 
Dev. 
 
Treatment Unblinded 
 
 
 
 
 
 
 
 
 
 
 
Other 
 
 
 
 
 
 
 
 
 
 
 


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14.1 
Description of Study Subjects 
14.1.1 Disposition of Subjects 
Table 9: 
Subject Disposition by Treatment Group, Amebiasis Group, Asymptomatic Population 
This table will be repeated for the symptomatic population. 
Subject 
Disposition  
Auranofin, 6 mg  
(N=X) 
Placebo 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
% 
n 
% 
Screened 
x 
x.x 
x 
x.x 
x 
x.x 
Enrolled and Received Treatment 
 
 
 
 
 
 
Received dose 1 
 
 
 
 
 
 
Received dose 2 
 
 
 
 
 
 
Received dose 3 
 
 
 
 
 
 
Received dose 4 
 
 
 
 
 
 
Received dose 5 
 
 
 
 
 
 
Received dose 6 
 
 
 
 
 
 
Received dose 7 
 
 
 
 
 
 
Received All Scheduled Doses a 
 
 
 
 
 
 
Completed Final Stool Sample (Day 7)  
 
 
 
 
 
 
Completed Follow-up (Study Day 28)a 
 
 
 
 
 
 
Completed Per Protocolb 
 
 
 
 
 
 
aRefer to Listing 16.2.1 for reasons subjects discontinued or terminated early. 
bRefer to Listing 16.2.2.1 for protocol deviations and Listing 16.2.3 for reasons subjects are excluded from the per protocol 
population. 
 


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Table 10: 
Subject Disposition by Treatment Group, Giardiasis Group, Asymptomatic Population 
This table will be repeated for the symptomatic population. 
Subject 
Disposition  
Auranofin, 6 mg  
(N=X) 
Placebo 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
% 
n 
% 
Screened 
x 
x.x 
x 
x.x 
x 
x.x 
Enrolled and Received Treatment 
 
 
 
 
 
 
Received dose 1 
 
 
 
 
 
 
Received dose 2 
 
 
 
 
 
 
Received dose 3 
 
 
 
 
 
 
Received dose 4 
 
 
 
 
 
 
Received dose 5 
 
 
 
 
 
 
Received All Scheduled Doses a 
 
 
 
 
 
 
Completed Final Stool Sample (Day 7)  
 
 
 
 
 
 
Completed Follow-up (Study Day 28)a 
 
 
 
 
 
 
Completed Per Protocolb 
 
 
 
 
 
 
aRefer to Listing 16.2.1 for reasons subjects discontinued or terminated early. 
bRefer to Listing 16.2.2.1 for protocol deviations and Listing 16.2.3 for reasons subjects are excluded from the per protocol 
population. 
 


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Table 11: 
Analysis Populations by Treatment Group, Amebiasis Group, Asymptomatic Population 
This table will be repeated for the symptomatic population. 
Analysis Populations 
Reason Subjects Excluded 
Auranofin  
(N=X) 
Placebo 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
n 
% 
n 
Modified Intent to Treat 
Population 
Did not receive study product 
x 
x.x 
x 
x.x 
x 
x.x 
Per-Protocol Population 
Any Reason 
 
 
 
 
 
 
 
Missed one or more doses of study 
product 
 
 
 
 
 
 
 
Out-of-window dose 
 
 
 
 
 
 
 
Out-of-window endpoint 
 
 
 
 
 
 
 
[Other reasons may be specified] 
 
 
 
 
 
 
Safety Population 
Any Reason 
 
 
 
 
 
 
 
[Reason 1] 
 
 
 
 
 
 
 
[Reason 2] 
 
 
 
 
 
 
Table 12: 
Analysis Populations by Treatment Group, Giardiasis Group 
Table will be formatted as Table 11. 
This table will be repeated for the symptomatic population. 
 


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Table 13: 
Dates of First Treatment by Disease Group and Treatment Group, Asymptomatic Population 
This table will be repeated for the symptomatic population. 
Dates of Dosing 
Amebiasis Group, 
Auranofin, 6 mg 
(N=X) 
Amebiasis Group, 
Placebo 
(N=X) 
Giardiasis Group, 
Auranofin, 6 mg 
(N=X) 
Giardiasis Group, 
Placebo 
(N=X) 
 
All Subjects 
(N=X) 
DDMMMYYYY-DDMMMYYYY 
[categorize based on length of enrollment period] 
x 
x 
x 
x 
x 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Total 
 
 
 
 
 
 


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Table 14: 
Ineligibility Summary of Screen Failures, Asymptomatic Population 
This table will be repeated for the symptomatic population, which had different inclusion and exclusion criteria. 
Inclusion/Exclusion Category 
Inclusion/Exclusion Criterion 
Number of Times Item Marked 
Ineligible* 
Inclusion and Exclusion 
Number of subjects failing any eligibility criterion 
x 
Inclusion 
Provide written informed consent prior to initiation of any 
study procedures. 
x 
Inclusion 
Able to understand and comply with planned study 
procedures and be available for all study visits. 
x 
Inclusion 
Male or non-pregnant female 18-65 years of age, inclusive. 
x 
Inclusion 
Amebiasis or giardiasis identified by positive Tech Lab 
antigen test, positive wet mount or trichome (confirmed by 
PCR), or positive stool PCR for E. histolytica or Giardia. If 
a subject is infected with both E. histolytica and Giardia, 
they will be enrolled in the E. histolytica study arm. Once 
the Entamoeba study arm is fully enrolled, any subsequent 
dual infected subjects will be enrolled in the Giardia arm.  . 
x 
Inclusion 
In otherwise good health.1 
x 
Inclusion 
Vital signs (oral temperature, pulse, and blood pressure) are 
all within normal protocol-defined ranges. 
x 
Inclusion 
Laboratory tests (blood urea nitrogen, creatinine, AST, 
ALT, white blood cells, platelets, hemoglobin, and 
hematocrit) are all within normal protocol-defined reference 
ranges.2 
x 
                                                 
1 As determined by medical history and targeted physical examination, if indicated based on medical history, to 
evaluate acute or currently ongoing chronic medical diagnoses or conditions that would affect the assessment of 
eligibility and safety of subjects. Existing medical diagnoses or conditions except those in the Subject Exclusion 
Criteria) must be deemed as stable chronic medical conditions.  A stable chronic medical condition is defined as no 
change in prescription medication, dose, or frequency of medication in the last 3 months (90 days) and health outcomes 
of the specific disease are considered to be within acceptable limits in the last 6 months (180 days).  Any change due to 
change of health care provider, insurance company, or that is done for financial reasons, as long as in the same class of 
medication, will not be considered a violation of this inclusion criterion.  Any change in prescription medication due to 
improvement of a disease outcome, as determined by the site principal investigator or appropriate sub-investigator, will 
not be considered a violation of this inclusion criterion.  Subjects may be on chronic or as needed (prn) medications if, 
in the opinion of the site principal investigator or appropriate sub-investigator, they pose no additional risk to subject 
safety. Topical, nasal, and inhaled medications, vitamins, and contraceptives are permitted. 
2 The normal protocol-defined ranges for laboratory tests include (a) blood urea nitrogen (BUN) between 5 and 24 
mg/dL, inclusive, (b) creatinine between 53  and 106 umol/L, inclusive, males and between 44  and 97umol/L, 
inclusive, for females, (c) AST of 38 U/L or less for males and 32 U/L or less for females, (d) ALT of 41.0 U/L or 
lessor males and 31 U/L or less for females, (e) WBC between 3.5 and 11.0 109/L, inclusive, (f) platelets between 150 
and 450 109/L, inclusive, (g) hemoglobin between 12.5 and 17.5 gm/dL, inclusive, for males and 11.5 and 16.5 gm/dL, 
 


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Table 14: 
Ineligibility Summary of Screen Failures, Asymptomatic Population (continued) 
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Inclusion/Exclusion Category 
Inclusion/Exclusion Criterion 
Number of Times Item Marked 
Ineligible* 
Inclusion 
Urinalysis with no greater than trace protein. If a high 
protein is confirmed to be due to menstruation, it should be 
repeated 
x 
Inclusion 
Women of reproductive potential3 must have a negative 
urine pregnancy test within 72 hours of starting study 
medications.   
x 
Inclusion 
Female subjects who are participating in sexual activity that 
could lead to pregnancy must agree to use highly effective4 
contraception plus condoms while receiving auranofin and 
for 15 weeks after.  These will be supplied to female 
subjects while they are on the trial and for the follow-up 
period of 4 months total.  Females on effective forms of 
birth control will continue while on the study and for the 
follow-up period of 4 months total. 
 
x 
Exclusion 
Known intolerance of auranofin or gold compounds. 
x 
Exclusion 
Pregnant or breastfeeding women or women who plan to 
become pregnant or breastfeed at any given time during the 
study or within 2 months of study completion. 
x 
Exclusion 
Diarrhea (> 3 loose stools/ 24 hours) within 7 days prior to 
starting study medications. 
x 
Exclusion 
Anticipated travel of more than 50 km from study site 
planned in next month. 
x 
Exclusion 
Evidence of clinically significant renal, hepatic, or 
immunologic impairment.5 
x 
Exclusion 
Current use of systemic antibiotics or metronidazole. 
x 
                                                 
inclusive, for females, (h) hematocrit between 40.0 and 52.0%, inclusive, for males and 35.0 and 47.0%, inclusive, for 
females. Subjects with low BUN or creatinine will be allowed to enroll, as it is not of clinical significance. 
3 Female subjects who are surgically sterile via tubal sterilization, bilateral oophorectomy or hysterectomy or who have 
been postmenopausal for greater than 1 year are not considered to be of reproductive potential. 
4 Highly effective methods of contraception are defined as having low failure rates (i.e. less than 1% per year) when 
used consistently and correctly and may include, but are not limited to, abstinence from intercourse, monogamous 
relationship with a vasectomized partner, male condoms or diaphragm with spermicide, intrauterine devices, and 
licensed hormonal methods. 
5 Renal: Creatinine >1.5 mg/dL, Blood urea nitrogen >30 mg/dL, or urine dipstick 1+ protein; Hepatic: AST or ALT 
>60 U/L; Immunologic: White blood cells < 4.0 (109/L), Platelets < 100 (106/L), or Hemoglobin <11 gm/dL, 
Hematocrit <30% 


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Table 14: 
Ineligibility Summary of Screen Failures, Asymptomatic Population (continued) 
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Inclusion/Exclusion Category 
Inclusion/Exclusion Criterion 
Number of Times Item Marked 
Ineligible* 
Exclusion 
Has any condition that would, in the opinion of the site 
investigator, place the subject at an unacceptable risk of 
injury or render the subject unable to meet the requirements 
of the protocol. 
x 
Exclusion 
Concurrent participation in other investigational protocols or 
receipt of an investigational product within the previous 30 
days. 
x 
Exclusion 
History of alcohol or drug abuse within the last five years. 
X 
Exclusion 
Current use of systemic antibiotics or metronidazole. 
x 
*More than one criterion may be marked per subject. 
 


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14.1.2 Demographic Data by Study Group 
Table 15: 
Distribution of Sex, Ethnicity, and Race by Treatment Group, Asymptomatic Population 
This table will be repeated for the symptomatic population, but will also split out the summaries by site. 
Demographic 
Category 
Characteristic 
Amebiasis Group, 
Auranofin, 
6 mg  
(n=34) 
Amebiasis Group, 
Placebo  
(n=34) 
Amebiasis Group, All 
subjects 
(n=68) 
Giardiasis Group, 
Auranofin, 6 mg  
(n=34) 
Giardiasis 
Group,  
Placebo  
(n=34) 
Giardiasis 
Group,  
All subjects 
(n=68) 
 
All 
subjects 
(n=136) 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Sex  
Male 
x 
x.x 
 
 
 
 
 
 
 
 
 
 
 
 
 
Female 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Ethnicity  
Not Hispanic or Latino 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Hispanic or Latino 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Not Reported 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Unknown 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Race  
American Indian or Alaska 
Native 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Asian 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Native Hawaiian or Other 
Pacific Islander 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Black or African American 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
White 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Multi-Racial 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Unknown 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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Table 16: 
Age Statistics (in Years) by Disease Group and Treatment Group, Asymptomatic Population 
This table will be repeated for the symptomatic population, but will also split out the summaries by site. 
Variable 
Statistic 
Amebiasis Group, 
Auranofin, 6 mg  
(n=34) 
Amebiasis Group, 
Placebo  
(n=34) 
Amebiasis Group, 
All subjects 
(n=68) 
Giardiasis Group, 
Auranofin, 6 mg  
(n=34) 
Giardiasis Group,  
Placebo  
(n=34) 
Giardiasis Group,  
All subjects 
(n=68) 
All subjects 
(n=136) 
Age 
Mean 
x.x 
x.x 
x.x 
 
 
 
 
 
Standard 
Deviation 
x.x 
x.x 
x.x 
 
 
 
 
 
Median 
x.x 
x.x 
x.x 
 
 
 
 
 
Minimum 
x 
x 
x 
 
 
 
 
 
Maximum 
x 
x 
x 
 
 
 
 
 


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14.1.3 Prior and Concurrent Medical Conditions 
Table 17: 
Distribution of Subjects with Pre-Existing Medical Conditions by MedDRA 
System Organ Class, Disease Group, and Treatment Group, Asymptomatic Population 
This table will be repeated for the symptomatic population. 
 
Amebiasis Group 
Giardiasis Group 
All Subjects, 
(both disease 
groups) 
(N=X) 
MedDRA System Organ Class 
Auranofin, 6 
mg 
(N=X) 
Placebo 
(N=X) 
All Subjects 
(N=X) 
Auranofin, 6 
mg 
(N=X) 
Placebo 
(N=X) 
All 
Subjects 
(N=X) 
n 
% 
n 
% 
 
 
 
 
 
 
Any SOC 
x 
x.x 
x 
x.x 
 
 
 
 
 
 
[SOC 1] 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: This table presents number and percentage of subjects.  A subject is only counted once per SOC. 
 


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Section 14.2 Efficacy Data 
Table 18: 
Comparison of Clearance Rates between Auranofin Group and Placebo Group, Day 7 (primary endpoint), 
Amebiasis Group, Asymptomatic Population 
 
Number of 
Subjects 
Number with 
Parasitological 
Response by  
Day 7 
Clearance Rate 
(proportion with 
parasitological response)  
by Day 7 with 95% CI 
Difference in Clearance 
Rate between Auranofin 
and Placebo with 95% 
CI 
Odds Ratio with 
95% CI 
Adjusted 
Odds 
Ratio with 
95% CI 
P-value 
from 
Fisher’s 
exact test 
Auranofin, 6 mg 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
For the asymptomatic population, parasitological response is defined as no detection of cysts or trophozoites by microscopic exam or negative antigen detection. 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
A footnote will list the covariates that were included in the model for the adjusted odds ratio.  
Table 19: 
Comparison of Clearance Rates between Auranofin Group and Placebo Group, Day 5 (primary endpoint), 
Giardiasis Group, Asymptomatic Population 
 
Number of 
Subjects 
Number with 
Parasitological 
Response by  
Day 5 
Clearance Rate 
(proportion with 
parasitological response)  
by Day 5 with 95% CI 
Difference in Clearance 
Rate between Auranofin 
and Placebo with 95% 
CI 
Odds Ratio with 
95% CI 
Adjusted 
Odds Ratio 
with 95% CI 
P-value 
from 
Fisher’s 
exact test 
Auranofin, 6 mg 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
For the asymptomatic population, parasitological response is defined as no detection of cysts or trophozoites by microscopic exam or negative antigen detection. 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
A footnote will list the covariates that were included in the model for the adjusted odds ratio. 
 


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Table 20: 
Comparison of Resolution of Diarrhea between Auranofin Group and Placebo Group, Day 7 (primary endpoint), 
Amebiasis Group, Symptomatic Population 
 
Number 
of 
Subjects 
Number with 
Resolution of 
Diarrhea  
Day 7 
Proportion with Resolution 
of Diarrhea  
by Day 7 with 95% CI 
Difference in 
proportions with 
95% CI 
Odds Ratio with 
95% CI 
Adjusted 
Odds Ratio 
with 95% CI 
P-value from 
Fisher’s exact 
test 
Auranofin, 6 mg 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
A footnote will list the covariates that were included in the model for the adjusted odds ratio. 
Table 21: 
Comparison of Resolution of Diarrhea between Auranofin Group and Placebo Group, Day 5 (primary endpoint), 
Giardiasis Group, Symptomatic Population 
 
Number of 
Subjects 
Number with 
Resolution of 
Diarrhea  
Day 5 
Proportion with Resolution 
of Diarrhea  
by Day 5 with 95% CI 
Difference in 
proportions with 
95% CI 
Odds Ratio 
with 95% CI 
Adjusted 
Odds Ratio 
with 95% CI 
P-value from 
Fisher’s exact 
test 
Auranofin, 6 mg 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here.  
A footnote will list the covariates that were included in the model for the adjusted odds ratio. 
 


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Table 22: 
Comparison of Clearance Rates between Auranofin Group and Placebo Group, Asymptomatic Amebiasis Group 
 
Clearance rate (proportion with parasitological response) by the specified day (95% CI) 
 
Number of Subjects 
Day 3 
Day 5 
Day 7 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% 
CI) 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
In the asymptomatic population, parasitological response is defined as no detection of cysts or trophozoites by microscopic exam or negative antigen detection. 
Footnote 1: The method for CI calculation will be denoted here. 
Table 23: 
Comparison of Clearance Rates between Auranofin Group and Placebo Group, Asymptomatic Giardiasis Group 
 
 
Clearance rate (proportion with parasitological response) by the specified day (95% CI) 
 
Number of Subjects 
Day 3 
Day 5 
Auranofin, 6 mg 
 
 
 
Placebo 
 
 
 
Difference in clearance rates 
(95% CI) 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
In the asymptomatic population, parasitological response is defined as no detection of cysts or trophozoites by microscopic exam or negative antigen detection. 
Footnote 1: The method for CI calculation will be denoted here 
 
 


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Table 24: 
Comparison of Clearance Rates between Auranofin Group and Placebo Group, Symptomatic Amebiasis Group 
 
Clearance rate (proportion with parasitological response) by the specified day (95% 
CI) 
 
Number of Subjects 
Day 3 
Day 5 
Day 7 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% CI) 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
In the symptomatic population, parasitological response is defined as no detection of trophozoites by microscopic exam. 
Footnote 1: The method for CI calculation will be denoted here. 
Table 25: 
Comparison of Clearance Rates between Auranofin Group and Placebo Group, Symptomatic Giardiasis Group 
 
 
Clearance rate (proportion with parasitological response) by the specified day (95% CI) 
 
Number of Subjects 
Day 3 
Day 5 
Auranofin, 6 mg 
 
 
 
Placebo 
 
 
 
Difference in clearance rates 
(95% CI) 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
In the symptomatic population, parasitological response is defined as no detection of trophozoites by microscopic exam. 
Footnote 1: The method for CI calculation will be denoted here 
 


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Table 26: 
Mixed Effects Model Coefficient Estimates, Asymptomatic Amebiasis Group 
Coefficient 
Estimate 
Standard Error 
P-value 
β0: Intercept 
 
 
 
β1: Treatment 
 
 
 
β2: Day 3 
 
 
 
β3: Day 3 * Treatment 
 
 
 
β4: Day 5 
 
 
 
β5: Day 5 * Treatment 
 
 
 
β6: Day 7 
 
 
 
β7: Day 7 * Treatment 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Table will be repeated for the symptomatic population 
. 
 


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Table 27: 
Mixed Effects Model Coefficient Estimates, Asymptomatic Giardiasis Group 
Coefficient 
Estimate 
 
Standard Error 
P-value 
β0: Intercept 
 
 
 
β1: Treatment 
 
 
 
β2: Day 3 
 
 
 
β3: Day 3 * Treatment 
 
 
 
β4: Day 5 
 
 
 
β5: Day 5 * Treatment 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Table will be repeated for the symptomatic population. 
 


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Table 28: 
Comparison of Proportion of Subjects with Negative Stool Antigen Test between Auranofin Group and Placebo Group, 
Asymptomatic Amebiasis Group 
 
Proportion with negative stool antigen tests (95% CI)  
on the specified day 
 
Number of 
Subjects 
During 
Screening 
Day 1 
Day 3 
Day 5 
Day 7 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
Difference in 
clearance rates (95% 
CI) 
 
 
 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
Table will be repeated for the symptomatic population 
 
 


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Table 29: 
Comparison of Proportion of Subjects with Sustained Cure between Auranofin Group and Placebo Group, 
Asymptomatic Amebiasis Group 
 
Proportion with sustained cure on the specified day and 95% CI  
 
Number of Subjects 
Day 7 
Day 14 
Day 28 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates 
(95% CI) 
 
 
 
 
The following definitions are for the asymptomatic amebiasis group: 
Sustained cure on Day 14 is defined as no detection of cysts or trophozoites on microscopic exam or negative antigen detection on Days 7 and 14. 
Sustained cure on Day 28 is defined to be no detection of cysts or trophozoites on microscopic exam or negative antigen detection on Days 7, 14, and 28. 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
Table will be repeated for the symptomatic population. 
Footnotes for the repeat table: 
Sustained cure on Day 14 is defined to be no detection of trophozoites on microscopic exam on Days 7 and 14. 
Sustained cure on Day 28 is defined to be no detection of trophozoites on microscopic exam on Days 7, 14, and 28. 
 


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Table 30: 
Comparison of Proportion of Subjects with Sustained Cure between Auranofin Group and Placebo Group, 
Asymptomatic Giardiasis Group 
 
Proportion with sustained cure on the specified day and 95% CI  
 
Number of Subjects 
Day 5 
Day 14 
Day 28 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% CI) for the difference 
 
 
 
The following definitions are for the asymptomatic giardiasis group: 
Sustained cure on Day 14 is defined to be no detection of cysts or trophozoites on microscopic exam or negative antigen detection on Days 5 and 14. 
Sustained cure on Day 28 is defined to be no detection of cysts or trophozoites on microscopic exam or negative antigen detection on Days 5, 14, and 28. 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
Table will be repeated for the symptomatic population. 
Footnotes for the repeat table: 
Sustained cure on Day 14 is defined to be no detection of trophozoites on microscopic exam on Days 5 and 14. 
Sustained cure on Day 28 is defined to be no detection of trophozoites on microscopic exam on Days 5, 14, and 28. 
 
 


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Table 31: 
Comparison of Proportion of Subjects with Relapse (Same Strain), and/or Reinfection (Different Strain) by Day 14, - 
Asymptomatic Amebiasis Group 
 
Number of Subjects 
Proportion with Relapse by 
Day 14  
(95% CI) 
Proportion with Reinfection by 
Day 14 
(95% CI) 
Proportion with Relapse and/or 
Reinfection by Day 14 
(95% CI) 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% 
CI) 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
Table 32: 
Comparison of Proportion of Subjects with Relapse (Same Strain), and/or Reinfection (Different Strain) by Day 14, - 
Asymptomatic Giardiasis Group 
 
Number of Subjects 
Proportion with Relapse by 
Day 14  
(95% CI) 
Proportion with Reinfection by 
Day 14 
(95% CI) 
Proportion with Relapse and/or 
Reinfection by Day 14 
(95% CI) 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% 
CI) 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
Tables will be repeated for the symptomatic population. 
 
 


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Table 33: 
Comparison of Proportion of Subjects with Relapse (Same Strain), and/or Reinfection (Different Strain) by Day 28, -
Asymptomatic Amebiasis Group 
 
Number of 
Subjects 
Proportion with Relapse by 
Day 28  
(95% CI) 
Proportion with Reinfection by 
Day 28 
(95% CI) 
Proportion with Relapse and/or 
Reinfection by Day 28 
(95% CI) 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% 
CI) 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here. 
Table 34: 
Comparison of Proportion of Subjects with Relapse (Same Strain), and/or Reinfection (Different Strain) by Day 28, -
Asymptomatic Giardiasis Group 
 
Number of 
Subjects 
Proportion with Relapse by 
Day 28  
(95% CI) 
Proportion with Reinfection by 
Day 28 
(95% CI) 
Proportion with Relapse and/or 
Reinfection by Day 28 
(95% CI) 
Auranofin, 6 mg 
 
 
 
 
Placebo 
 
 
 
 
Difference in clearance rates (95% CI) 
 
 
 
 
Note: These analyses will be performed both on the m-ITT population and the per-protocol population. 
Footnote 1: The method for CI calculation will be denoted here 
Tables will be repeated for the symptomatic population. 
 


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14.3 
Safety Data 
14.3.1 Displays of Adverse Events 
14.3.1.1 
Solicited Adverse Events 
Table 35: 
Number and Percentage of Subjects Experiencing Solicited Events with 95% Confidence Intervals by Symptom 
and Treatment Group, - Asymptomatic Amebiasis Group 
Symptom 
Auranofin, 6 mg 
(N=X) 
Placebo 
(N=X) 
n 
% 
95% CI 
n 
% 
95% CI 
Any Symptom 
x 
x.x 
x.x, x.x 
x 
x.x 
x.x, x.x 
Loose stools or diarrhea 
 
 
 
 
 
 
Abdominal Pain 
 
 
 
 
 
 
Nausea with or without vomiting 
 
 
 
 
 
 
Constipation 
 
 
 
 
 
 
Anorexia 
 
 
 
 
 
 
Flatulence 
 
 
 
 
 
 
Dyspepsia 
 
 
 
 
 
 
Dysgeusia 
 
 
 
 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 35: 
Number and Percentage of Subjects Experiencing Solicited Events With 95% Confidence Intervals by Symptom 
and Treatment Group, - Asymptotic Amebiasis Group (continued) 
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Symptom 
Auranofin, 6 mg 
(N=X) 
Placebo 
(N=X) 
n 
% 
95% CI 
n 
% 
95% CI 
Pruritus 
 
 
 
 
 
 
Hair loss 
 
 
 
 
 
 
Urticaria 
 
 
 
 
 
 
Stomatitis 
 
 
 
 
 
 
Conjunctivitis 
 
 
 
 
 
 
Glossitis 
 
 
 
 
 
 
Hematuria 
 
 
 
 
 
 
Rash 
 
 
 
 
 
 
Table will be repeated for the symptomatic study population but will exclude loose stools or diarrhea. 
 


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Table 36: 
Number and Percentage of Subjects Experiencing Solicited Events with 95% Confidence Intervals by Symptom and Treatment 
Group, Asymptomatic Giardiasis Group 
Symptom 
Auranofin, 6 mg 
(N=X) 
Placebo 
(N=X) 
n 
% 
95% CI 
n 
% 
95% CI 
Any Symptom 
x 
x.x 
x.x, x.x 
x 
x.x 
x.x, x.x 
Loose stools or diarrhea 
 
 
 
 
 
 
Abdominal Pain 
 
 
 
 
 
 
Nausea with or without vomiting 
 
 
 
 
 
 
Constipation 
 
 
 
 
 
 
Anorexia 
 
 
 
 
 
 
Flatulence 
 
 
 
 
 
 
Dyspepsia 
 
 
 
 
 
 
Dysgeusia 
 
 
 
 
 
 
Pruritus 
 
 
 
 
 
 
Hair loss 
 
 
 
 
 
 
Urticaria 
 
 
 
 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 36: 
Number and Percentage of Subjects Experiencing Solicited Events with 95% Confidence Intervals by Symptom and Treatment 
Group, Asymptomatic Giardiasis Group 
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Symptom 
Auranofin, 6 mg 
(N=X) 
Placebo 
(N=X) 
n 
% 
95% CI 
n 
% 
95% CI 
Stomatitis 
 
 
 
 
 
 
Conjunctivitis 
 
 
 
 
 
 
Glossitis 
 
 
 
 
 
 
Hematuria 
 
 
 
 
 
 
Rash 
 
 
 
 
 
 
N = Number of subjects in the Safety Analysis Population who received the specified dose. Severity is the maximum severity reported. 
Table will be repeated for the symptomatic study population but will exclude loose stools or diarrhea. 
 


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14.3.1.2 
Unsolicited Adverse Events 
Table 37: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA System 
Organ Class and Preferred Term, Maximum Severity, Relationship, Asymptomatic Amebiasis Group, Auranofin, 6 mg (n=x) 
MedDRA System Organ Class 
MedDRA Preferred Term 
Any Incidence 
Severity [1] 
Relationship to Treatment [2] 
Mild 
Moderate 
Severe 
Not Related 
Related 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
Any PT 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
[SOC 1] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of subjects in the Safety Analysis Population.  
[1] For severity, a subject is counted once per preferred term and is summarized according to their highest severity. 
[2] For relationship, a subject is only counted once per preferred term and is summarized according to their closest relationship. 
Table will be repeated for the symptomatic study population. 
 


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Table 38: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA System 
Organ Class and Preferred Term, Maximum Severity, Relationship, Asymptomatic Amebiasis Group, Placebo (n=x) 
MedDRA System Organ Class 
MedDRA Preferred Term 
Any Incidence 
Severity [1] 
Relationship to Treatment [2] 
Mild 
Moderate 
Severe 
Not Related 
Related 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
Any PT 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
[SOC 1] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of subjects in the Safety Analysis Population.  
[1] For severity, a subject is counted once per preferred term and is summarized according to their highest severity. 
[2] For relationship, a subject is only counted once per preferred term and is summarized according to their closest relationship. 
Table will be repeated for the symptomatic study population. 
 


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Table 39: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA System Organ Class and Preferred 
Term, Maximum Severity, Relationship, Asymptomatic Giardiasis Group, Auranofin, 6 mg (n=x) 
MedDRA System Organ Class 
MedDRA Preferred Term 
Any Incidence 
Severity [1] 
Relationship to Treatment [2] 
Mild 
Moderate 
Severe 
Not Related 
Related 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
Any PT 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
[SOC 1] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of subjects in the Safety Analysis Population.  
[1] For severity, a subject is counted once per preferred term and is summarized according to their highest severity. 
[2] For relationship, a subject is only counted once per preferred term and is summarized according to their closest relationship. 
Table will be repeated for the symptomatic study population. 
 


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Table 40: 
Number and Percentage of Subjects Experiencing Unsolicited Adverse Events by MedDRA System Organ Class and Preferred 
Term, Maximum Severity, Relationship, Asymptomatic Giardiasis Group, Placebo (n=x) 
MedDRA System Organ Class 
MedDRA Preferred Term 
Any Incidence 
Severity [1] 
Relationship to Treatment [2] 
Mild 
Moderate 
Severe 
Not Related 
Related 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Any SOC 
Any PT 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
[SOC 1] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
[SOC 2] 
Any PT 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 1] 
 
 
 
 
 
 
 
 
 
 
 
 
 
[PT 2] 
 
 
 
 
 
 
 
 
 
 
 
 
Note: N = Number of subjects in the Safety Analysis Population.  
[1] For severity, a subject is counted once per preferred term and is summarized according to their highest severity. 
[2] For relationship, a subject is only counted once per preferred term and is summarized according to their closest relationship. 
Table will be repeated for the symptomatic study population. 
 


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14.3.2 Listing of Deaths, Other Serious and Significant Adverse Events 
Table 41: 
Listing of Serious Adverse Events 
Listings will be sorted by study population, disease group, treatment group, subject ID 
Adverse 
Event 
Associated 
with Dose 
No. 
No. of Days 
Post 
Associated 
Dose 
(Duration) 
No. of Days 
Post Dose 
the Event 
Became 
Serious 
Reason 
Reported 
as an SAE 
Severity 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA 
System 
Organ 
Class 
MedDRA 
Preferred 
Term 
Subject ID: , Study Population: , Disease Group: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
Subject ID: , Study Population: , Disease Group: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 


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Table 42: 
Listing of Non-Serious, Unsolicited, Moderate or Severe Adverse Events, Asymptomatic Amebiasis Group 
Table will be repeated for the symptomatic population. 
Adverse 
Event 
Associated 
with Dose No. 
No. of Days 
Post 
Associated 
Dose 
(Duration) 
Severity 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action Taken 
with Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA 
System Organ 
Class 
MedDRA 
Preferred 
Term 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 


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Table 43: 
Listing of Non-Serious, Unsolicited, Moderate or Severe Adverse Events, Asymptomatic Giardiasis Group 
Table will be repeated for the symptomatic population. 
Adverse 
Event 
Associated 
with Dose No. 
No. of Days 
Post 
Associated 
Dose 
(Duration) 
Severity 
Relationship 
to Study 
Treatment 
If Not 
Related, 
Alternative 
Etiology 
Action Taken 
with Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA 
System Organ 
Class 
MedDRA 
Preferred 
Term 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
Subject ID: , Treatment Group: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 


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14.3.3  
Narratives of Deaths, Other Serious and Significant Adverse Events 
(not included in SAP, but this is a placeholder for the CSR) 
[This section of the clinical study report will include narrative text of deaths, serious, and significant adverse events.] 
 


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14.3.4 Abnormal Laboratory Value Listings (by Subject) 
Table 44: 
Listing of Abnormal Laboratory Results, Asymptomatic Amebiasis Group 
Subject 
ID 
Treatment 
Group 
Sex 
Age 
(years) 
Planned 
Time Point 
Actual 
Study Day 
Laboratory 
Parameter 
(Units) 
Result 
(Severity) 
Relationship to 
Treatment 
If Not Related, 
Alternate 
Etiology 
Action Taken 
with Study 
Treatment 
Subject 
Discontinued Due 
to Result? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Table 45: 
Listing of Abnormal Laboratory Results, Asymptomatic Giardiasis Group 
Subject 
ID 
Treatment 
Group 
Sex 
Age 
(years) 
Planned 
Time Point 
Actual 
Study Day 
Laboratory 
Parameter 
(Units) 
Result 
(Severity) 
Relationship to 
Treatment 
If Not Related, 
Alternate 
Etiology 
Action Taken 
with Study 
Treatment 
Subject 
Discontinued Due 
to Result? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 
 


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14.3.5 Displays of Laboratory Results 
Table 46: 
Laboratory Results by Parameter, Maximum Severity, Study Day, and Treatment Group, Asymptomatic Amebiasis Group -   
Any Laboratory Parameter 
Any laboratory 
parameter 
Treatment Group 
N 
None 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
Table 47: 
Laboratory Results by Parameter, Maximum Severity, Study Day, and Treatment Group, Asymptomatic Amebiasis Group - 
White Blood Cell Count 
White Blood Cell 
Count 
Treatment Group 
N 
None 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 
 


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Table 48: 
Laboratory Results by Parameter, Maximum Severity, Study Day, and Treatment Group, Asymptomatic Amebiasis Group - 
Hemoglobin 
Hemoglobin 
Treatment Group 
N 
None 
Mild/ 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
Table 49: 
Laboratory Results by Parameter, Maximum Severity, Study Day, and Treatment Group, Asymptomatic Amebiasis Group - 
Hematocrit 
Hematocrit 
Treatment Group 
N 
None 
Mild 
Grade 1 
Moderate 
Grade 2 
Severe 
Grade 3 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 


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Table 50: 
Laboratory Results by Parameter, Maximum Severity, Study Day, and Treatment Group, Asymptomatic Amebiasis Group - 
Platelets 
Platelets 
Treatment Group 
N 
None 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 


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Table 51: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and 
Treatment Group, Asymptomatic Amebiasis Group -White Blood Cell Count 
White Blood Cell 
Count 
Treatment Group 
N 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
Table 52: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and  
Treatment Group, Asymptomatic Amebiasis Group - Hemoglobin 
Hemoglobin 
Treatment Group 
N 
Mild/ 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 


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Table 53: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and  
Treatment Group, Asymptomatic Amebiasis Group - Hematocrit 
Hematocrit 
Treatment Group 
N 
Mild/ 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
Table 54: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and  
Treatment Group, Asymptomatic Amebiasis Group - Platelets 
Platelets 
Treatment Group 
N 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population 
 


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Table 55: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and Treatment Group, 
Asymptomatic Giardiasis Group -White Blood Cell Count 
White Blood Cell 
Count 
Treatment Group 
N 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
Table 56: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and Treatment Group, 
Asymptomatic Giardiasis Group - Hemoglobin 
Hemoglobin 
Treatment Group 
N 
Mild/ 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 
 


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Table 57: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and Treatment Group, 
Asymptomatic Giardiasis Group - Hematocrit 
Hematocrit 
Treatment Group 
N 
Mild/ 
Grade 1 
Moderate/ 
Grade 2 
Severe/ 
Grade 3 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
Table 58: 
Abnormal Laboratory Results Related to Study Treatment by Parameter, Maximum Severity, Study Day, and Treatment Group, 
Asymptomatic Giardiasis Group - Platelets 
Platelets 
Treatment Group 
N 
Mild/ 
Grade 1 
(Low) 
Mild/ 
Grade 1 
(High) 
Moderate/ 
Grade 2 
(Low) 
Moderate/ 
Grade 2 
(High) 
Severe/ 
Grade 3 
(Low) 
Severe/ 
Grade 3 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Repeat for symptomatic study population. 
 


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Table 59: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Amebiasis Group -  
White Blood Cells 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.xx 
xx.xx 
xx.x 
xx.x, xx.x 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest. 
Repeat for symptomatic study population. 
Table 60: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Amebiasis Group - Hemoglobin 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.xx 
xx.xx 
xx.x 
xx.x, xx.x 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest. 
Repeat for symptomatic study population. 
 
 


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Table 61: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Amebiasis Group -Hematocrit 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.x 
xx.x 
x 
xx, xx 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest. 
Repeat for symptomatic study population. 
Table 62: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Amebiasis Group - Platelets 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.xx 
xx.xx 
xx.x 
xx.x, xx.x 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest.  
Repeat for symptomatic study population. 
 


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Table 63: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Giardiasis Group -  
White Blood Cells 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.xx 
xx.xx 
xx.x 
xx.x, xx.x 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest.  
Repeat for symptomatic study population. 
Table 64: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Giardiasis Group - Hemoglobin 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.xx 
xx.xx 
xx.x 
xx.x, xx.x 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest.  
Repeat for symptomatic study population. 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 65: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Giardiasis Group - Hematocrit 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.x 
xx.x 
x 
xx, xx 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest.  
Repeat for symptomatic study population. 
Table 66: 
Laboratory Summary Statistics by Parameter, Study Day, and Treatment Group, Asymptomatic Giardiasis Group - Platelets 
Time Point 
Treatment Group 
N 
Mean 
Standard Deviation 
Median 
Min, Max 
Baseline 
Auranofin, 6 mg 
x 
xx.xx 
xx.xx 
xx.x 
xx.x, xx.x 
 
Placebo 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
Day 7, Change from Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
N = Number of subjects in the Safety population with non-missing values for the parameter of interest.  
Repeat for symptomatic study population. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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14.3.6 Displays of Vital Signs 
Table 67: 
Vital Signs by Assessment, Maximum Severity, Study Day, and 
Treatment Group, Amebiasis Group - Any Assessment 
This table series will be repeated for the symptomatic study population. 
Time Point 
Treatment 
Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity Post 
Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time 
point post baseline, including unscheduled assessments. N=Number of subjects in the Safety population with 
non-missing values. 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 68: 
Vital Signs by Assessment, Maximum Severity, Study Day, and 
Treatment Group, Amebiasis Group - Fever 
Time Point 
Treatment 
Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 
mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity Post Baseline 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post 
ncluding unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 69: 
Vital Signs by Assessment, Maximum Severity, Study Day, and 
Treatment Group, Amebiasis Group - Systolic Blood Pressure 
(Hypertension/Hypotension) 
Time 
Point 
Treatment 
Group 
N 
None 
Mild 
(Low) 
Mild 
(High) 
Moderate 
(Low) 
Moderate 
(High) 
Severe 
(Low) 
Severe 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline Auranofin, 
6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Max 
Severity 
Post 
Baseline 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time 
point post baseline, including unscheduled assessments. N=Number of subjects in the Safety population with 
non-missing values 
 


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Table 70: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group, 
Amebiasis Group - Diastolic Blood Pressure - Hypertension 
Time Point 
Treatment Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity 
Post Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post baseline, 
including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 71: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group, 
Amebiasis Group -Bradycardia 
Time Point 
Treatment Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity 
Post Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post baseline, 
including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 72: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group, 
Amebiasis Group - Tachycardia 
Time Point 
Treatment Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity 
Post Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post baseline, 
including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 73: 
Vital Signs by Assessment, Maximum Severity, Study Day, and 
Treatment Group, Giardiasis Group - Any Assessment 
This table series will be repeated for the symptomatic study population. 
Time Point 
Treatment 
Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 
mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity Post Baseline 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post 
baseline, including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 74: 
Vital Signs by Assessment, Maximum Severity, Study Day, and 
Treatment Group, Giardiasis Group - Fever 
Time Point 
Treatment 
Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 
mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity Post Baseline 
Auranofin, 6 
mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time 
point post baseline, including unscheduled assessments. N=Number of subjects in the Safety population with 
non-missing values. 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 75: 
Vital Signs by Assessment, Maximum Severity, Study Day, and 
Treatment Group, Giardiasis Group - Systolic Blood Pressure - 
(Hypertension/Hypotension) 
Time 
Point 
Treatment 
Group 
N 
None 
Mild 
(Low) 
Mild 
(High) 
Moderate 
(Low) 
Moderate 
(High) 
Severe 
(Low) 
Severe 
(High) 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline Auranofin, 
6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Max 
Severity 
Post 
Baseline 
Auranofin, 
6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time 
nt post baseline, including unscheduled assessments. N=Number of subjects in the Safety population with non-
sing values. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 76: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group, 
Giardiasis Group - Diastolic Blood Pressure - Hypertension 
Time Point 
Treatment Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity 
Post Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post baseline, 
including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 77: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group, 
Giardiasis Group - Bradycardia 
Time Point 
Treatment Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity 
Post Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post baseline, 
including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 78: 
Vital Signs by Assessment, Maximum Severity, Study Day, and Treatment Group, 
Giardiasis Group - Tachycardia 
Time Point 
Treatment Group 
N 
None 
Mild 
Moderate 
Severe 
Missing 
n 
% 
n 
% 
n 
% 
n 
% 
n 
% 
Baseline 
Auranofin, 6 mg 
x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
x 
x.x 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 1 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 2 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 3 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 4 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 5 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 6 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 7 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 14 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Day 28 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Max Severity 
Post Baseline 
Auranofin, 6 mg 
 
 
 
 
 
 
 
 
 
 
 
 
Placebo 
 
 
 
 
 
 
 
 
 
 
 
Note: The “Max Post Baseline” rows indicate the maximum severity experienced by each subject at any time point post baseline, 
including unscheduled assessments. N=Number of subjects in the Safety population with non-missing values. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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14.4 
Summary of Concomitant Medications 
Table 79: 
Number and Percentage of Subjects with Prior and Concurrent Medications by WHO 
Drug Classification and Treatment Group, Asymptomatic Amebiasis Group 
This table will be repeated for the symptomatic population. 
WHO Drug Code 
Level 1, Anatomic 
Group 
WHO Drug Code 
Level 2, Therapeutic 
Subgroup 
Auranofin, 6 mg 
(N=X) 
Placebo 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
% 
n 
% 
Any Level 1 Codes 
Any Level 2 Codes 
x 
x.x 
x 
x.x 
x 
x.x 
[ATC Level 1 - 1] 
Any [ATC 1 – 1] 
 
 
 
 
 
 
 
[ATC 2 - 1] 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
[ATC Level 1 – 2] 
[ATC 2 - 1] 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
N= Number of subjects in the Safety population with non-missing values 
. 
 


Statistical Analysis Plan - DMID Protocol: 15-0015 
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Table 80: 
Number and Percentage of Subjects with Prior and Concurrent Medications by WHO 
Drug Classification and Treatment Group, Asymptomatic Giardiasis Group 
This table will be repeated for the symptomatic population. 
WHO Drug Code 
Level 1, Anatomic 
Group 
WHO Drug Code 
Level 2, Therapeutic 
Subgroup 
Auranofin, 6 mg 
(N=X) 
Placebo 
(N=X) 
All Subjects 
(N=X) 
n 
% 
n 
% 
n 
% 
Any Level 1 Codes 
Any Level 2 Codes 
x 
x.x 
x 
x.x 
x 
x.x 
[ATC Level 1 - 1] 
Any [ATC 1 – 1] 
 
 
 
 
 
 
 
[ATC 2 - 1] 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
[ATC Level 1 – 2] 
[ATC 2 - 1] 
 
 
 
 
 
 
 
[ATC 2 - 2] 
 
 
 
 
 
 
 
[ATC 2 - 3] 
 
 
 
 
 
 
N= Number of subjects in the Safety population with non-missing values. 
 


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APPENDIX 2.  FIGURE MOCK-UPS 
 
 


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LIST OF FIGURES 
Figure 1: 
Schematic of Study Design, Part 1 ...........................................................................126 
Figure 2: 
Schematic of Study Design, Part 2 ...........................................................................127 
Figure 3: 
CONSORT Flow Diagram .......................................................................................128 
Figure 4: 
Kaplan-Meier Curves of Time to Diarrhea Resolution, Symptomatic 
Amebiasis Group ......................................................................................................129 
Figure 5: 
Maximum Severity of Solicited Symptoms per Subject by Day Post 
Treatment And Treatment Group, Asymptomatic Amebiasis Group (N=68) ..........130 
Figure 6: 
Frequency of Adverse Events by MedDRA System Organ Class and 
Severity Asymptomatic Amebiasis Group (N=68)...................................................131 
Figure 7: 
Frequency of Adverse Events by MedDRA System Organ Class and 
Severity Asymptomatic Giardiasis Group (N=68) ...................................................132 
Figure 8: 
Incidence of Adverse Events by MedDRA System Organ Class and Severity, 
Asymptomatic Amebiasis Group (N=68) .................................................................133 
Figure 9: 
Incidence of Adverse Events by MedDRA System Organ Class and Severity, 
Asymptomatic Giardiasis Group (N=68)..................................................................134 
Figure 10: Frequency of Unsolicited Adverse Events by Severity Asymptomatic 
Amebiasis Group (N=68) .........................................................................................135 
Figure 11: Frequency of Unsolicited Adverse Events by Severity Asymptomatic 
Giardiasis Group (N=68) ..........................................................................................136 
Figure 12: Incidence of Unsolicited Adverse Events by Asymptomatic Amebiasis 
Group (N=68) ...........................................................................................................137 
Figure 13: Incidence of Unsolicited Adverse Events by Severity Asymptomatic 
Giardiasis Group (N=68) ..........................................................................................138 
Figure 14: Frequency of Unsolicited Adverse Events by MedDRA System Organ Class 
and Relationship to Treatment - Asymptomatic Amebiasis Group (N=68) .............139 
Figure 15: Frequency of Unsolicited Adverse Events by MedDRA System Organ Class 
and Relationship to Treatment Asymptomatic Giardiasis Group (N=68) ................140 
Figure 16: Incidence of Unsolicited Adverse Events by MedDRA System Organ Class 
and Relationship to Treatment - Asymptomatic Amebiasis Group (N=68) .............141 
Figure 17: Incidence of Unsolicited Adverse Events by MedDRA System Organ Class 
and Relationship to Treatment - Asymptomatic Giardiasis Group (N=68) ..............142 
Figure 18: Frequency of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Amebiasis Group (N=68) .................................................................143 
Figure 19: Frequency of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Giardiasis Group (N=68)..................................................................144 


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Figure 20: Incidence of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Amebiasis Group (N=68) .................................................................145 
Figure 21: Incidence of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Giardiasis Group (N=68)..................................................................146 
Figure 22: Laboratory Results: Distribution on Baseline and Day 7 by Laboratory 
Parameter, Treatment Group, and Gender (where appropriate) - 
Asymptomatic Amebiasis Group (N=68) .................................................................147 
Figure 23: Laboratory Results: Distribution on Baseline and Day 7 by Laboratory 
Parameter, Treatment Group, and Gender (where appropriate) - 
Asymptomatic Giardiasis Group (N=68)..................................................................148 
 


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9.1 
Overall Study Design and Plan Description 
Figure 1: 
Schematic of Study Design, Part 1 
 


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Figure 2: 
Schematic of Study Design, Part 2 
 
 


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10.1 
Disposition of Subjects 
Figure 3: 
CONSORT Flow Diagram 
Assessed for 
Eligibility (n=x)
Excluded* (n=x)
♦   [Reason 1] (n=x)
♦   Concern of potential 
risk (n=x)
Test positive 
for amebiasis
Test positive 
for giardiasis
Allocated to Auranofin 
(n=x)
♦Received Auranofin (n=x)
♦Did not receive Auranofin 
(n=x)
Completed full course 
(n=x)
Completed partial course 
(n=x)
Completed full course 
(n=x)
Completed partial course 
(n=x)
Completed full course 
(n=x)
Completed partial course 
(n=x)
Allocated to Auranofin 
(n=x)
♦Received Auranofin (n=x)
♦Did not receive Auranofin 
(n=x)
Allocated to Placebo (n=x)
♦Received Placebo (n=x)
♦Did not receive Placebo 
(n=x)
Allocated to Placebo (n=x)
♦Received Placebo (n=x)
♦Did not receive Placebo 
(n=x)
Completed full course 
(n=x)
Completed partial course 
(n=x)
Analysed for Safety (n=x)
Excluded from Safety 
Analysis (n=x)
Included in mITT 
Population (efficacy 
analyses)  (n=x)
Included in per-Protocol 
Population (efficacy 
analyses) (n=x)
Randomized 
(n=x)
Randomized 
(n=x)
Analysed for Safety (n=x)
Excluded from Safety 
Analysis (n=x)
Included in mITT 
Population (efficacy 
analyses)  (n=x)
Included in per-Protocol 
Population (efficacy 
analyses) (n=x)
Analysed for Safety (n=x)
Excluded from Safety 
Analysis (n=x)
Included in mITT 
Population (efficacy 
analyses)  (n=x)
Included in per-Protocol 
Population (efficacy 
analyses) (n=x)
Analysed for Safety (n=x)
Excluded from Safety 
Analysis (n=x)
Included in mITT 
Population (efficacy 
analyses)  (n=x)
Included in per-Protocol 
Population (efficacy 
analyses) (n=x)
 
 


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14.2.2 
Efficacy/Immunogenicity Response Figures by Measure, Treatment/Vaccination, and Time 
Point 
Figure 4: 
Kaplan-Meier Curves of Time to Diarrhea Resolution, Symptomatic Amebiasis Group 
Figure will be repeated for the symptomatic giardiasis group. 
 
 
This figure will display Kaplan-Meier curves for 
treatment and control arms with the log rank 
p-value. 


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14.3.1.1 
Solicited Adverse Events 
Figure 5: 
Maximum Severity of Solicited Symptoms per Subject by Day Post Treatment 
And Treatment Group, Asymptomatic Amebiasis Group (N=68) 
Figure will be repeated for the asymptomatic giardiasis, symptomatic amebiasis, and symptomatic giardiasis groups. 
 
 


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Figure 6: 
Frequency of Adverse Events by MedDRA System Organ Class and Severity 
Asymptomatic Amebiasis Group (N=68) 
Figure will be repeated for the symptomatic amebiasis group. 
 
 


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14.3.1.2 Unsolicited Adverse Events 
Figure 7: 
Frequency of Adverse Events by MedDRA System Organ Class and Severity 
Asymptomatic Giardiasis Group (N=68) 
Figure will be repeated for the symptomatic giardiasis group. 
 
 


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Figure 8: 
Incidence of Adverse Events by MedDRA System Organ Class and Severity, 
Asymptomatic Amebiasis Group (N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 


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Figure 9: 
Incidence of Adverse Events by MedDRA System Organ Class and Severity, 
Asymptomatic Giardiasis Group (N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
 
 


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Figure 10: 
Frequency of Unsolicited Adverse Events by Severity Asymptomatic Amebiasis Group 
(N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 
 


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Figure 11: 
Frequency of Unsolicited Adverse Events by Severity Asymptomatic Giardiasis Group 
(N=68)  
Figure will be repeated for the symptomatic giardiasis group 
 


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Figure 12: 
Incidence of Unsolicited Adverse Events by Asymptomatic Amebiasis Group (N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 
 


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Figure 13: 
Incidence of Unsolicited Adverse Events by Severity Asymptomatic Giardiasis Group 
(N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
 
 


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Figure 14: 
Frequency of Unsolicited Adverse Events by MedDRA System Organ Class and 
Relationship to Treatment - Asymptomatic Amebiasis Group (N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 
 


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Figure 15: 
Frequency of Unsolicited Adverse Events by MedDRA System Organ Class and 
Relationship to Treatment Asymptomatic Giardiasis Group (N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
 
 


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Figure 16: 
Incidence of Unsolicited Adverse Events by MedDRA System Organ Class and 
Relationship to Treatment - Asymptomatic Amebiasis Group (N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 
 


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Figure 17: 
Incidence of Unsolicited Adverse Events by MedDRA System Organ Class and 
Relationship to Treatment - Asymptomatic Giardiasis Group (N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
 
 


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Figure 18: 
Frequency of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Amebiasis Group (N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 
 


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Figure 19: 
Frequency of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Giardiasis Group (N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
 
 


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Figure 20: 
Incidence of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Amebiasis Group (N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
 
 


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Figure 21: 
Incidence of Unsolicited Adverse Events by Relationship to Treatment -  
Asymptomatic Giardiasis Group (N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
 
 


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14.3.5 
Displays of Laboratory Results 
Figure 22: 
Laboratory Results: Distribution on Baseline and Day 7 by Laboratory Parameter, 
Treatment Group, and Gender (where appropriate) - Asymptomatic Amebiasis Group 
(N=68)  
Figure will be repeated for the symptomatic amebiasis group. 
 
Grading criteria were changed during the course of the study.  The most conservative grading thresholds will 
be displayed.  Other ranges will be shown as dashed lines or indicated in a footnote. 
 
 


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Figure 23: 
Laboratory Results: Distribution on Baseline and Day 7 by Laboratory Parameter, 
Treatment Group, and Gender (where appropriate) - Asymptomatic Giardiasis Group 
(N=68)  
Figure will be repeated for the symptomatic giardiasis group. 
 
Grading criteria were changed during the course of the study.  The most conservative grading thresholds will 
be displayed.  Other ranges will be shown as dashed lines or indicated in a footnote. 
 


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APPENDIX 3. LISTINGS MOCK-UPS 
 
 


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LISTINGS 
Listing 1: 16.2.1.1: Early Terminations or Discontinued Subjects - Asymptomatic 
Amebiasis Group ......................................................................................................153 
Listing 2 : 16.2.1.2: Early Terminations or Discontinued Subjects - Asymptomatic 
Giardiasis Group .......................................................................................................153 
Listing 3:  16.2.2.1: Subject-Specific Protocol Deviations -Asymptomatic Amebiasis 
Group ........................................................................................................................154 
Listing 4: 16.2.2.2: Subject-Specific Protocol Deviations - Asymptomatic Giardiasis 
Group ........................................................................................................................154 
Listing 5: 16.2.2.2.3: Non-Subject-Specific Protocol Deviations -Asymptomatic 
Amebiasis Group ......................................................................................................155 
Listing 6:  16.2.2.2.4: Non-Subject-Specific Protocol Deviations - Asymptomatic 
Giardiasis Group .......................................................................................................155 
Listing 7:  16.2.3.1: Subjects Excluded from Analysis Populations - Asymptomatic 
Amebiasis Group ......................................................................................................156 
Listing 8: 16.2.3.2: Subjects Excluded from Analysis Populations - Asymptomatic 
Giardiasis Group .......................................................................................................156 
Listing 9:  16.2.4.1: Demographic Data - Asymptomatic Amebiasis Group .............................157 
Listing 10:  16.2.4.2: Demographic Data - Asymptomatic Giardiasis Group ..............................157 
Listing 11: 16.2.4.3: Pre-Existing Medical Conditions - Asymptomatic Amebiasis 
Group ........................................................................................................................158 
Listing 12: 16.2.4.4: Pre-Existing Medical Conditions - Asymptomatic Giardiasis Group .......158 
Listing 13: 16.2.5.1: Compliance and/or Drug Concentration Data - Asymptomatic 
Amebiasis Group ......................................................................................................159 
Listing 14: 16.2.5.2: Compliance and/or Drug Concentration Data - Asymptomatic 
Giardiasis Group .......................................................................................................159 
Listing 15: 16.2.6.1: Listing of Efficacy Endpoints - Asymptomatic Amebiasis Group ............160 
Listing 16: 16.2.6.2: Listing of Efficacy Endpoints - Symptomatic Amebiasis Group ..............160 
Listing 17: 16.2.6.3: Listing of Efficacy Endpoints - Asymptomatic Giardiasis Group ............161 
Listing 18:  16.2.6.4: Listing of Efficacy Endpoints - Symptomatic Giardiasis Group ...............161 
Listing 19: 16.2.7.1.1: Solicited Events - Asymptomatic Amebiasis Group – First listing ........162 
Listing 20: 16.2.7.1.2: Solicited Events - Asymptomatic Amebiasis Group – Second 
listing ........................................................................................................................162 
Listing 21: 16.2.7.1.2: Solicited Events - Asymptomatic Giardiasis Group – First listing ........163 
Listing 22: 16.2.7.1.3: Solicited Events Asymptomatic Giardiasis Group – Second 
listing ........................................................................................................................163 


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Listing 23: 16.2.7.2.1: Unsolicited Adverse Events - Asymptomatic Amebiasis Group ............164 
Listing 24: 16.2.7.2.2: Unsolicited Adverse Events - Asymptomatic Giardiasis Group ............165 
Listing 25: 16.2.8.1.1: Clinical Laboratory Results: Hematology - Asymptomatic 
Amebiasis Group ......................................................................................................166 
Listing 26: 16.2.8.1.2: Clinical Laboratory Results: Hematology - Asymptomatic 
Giardiasis Group .......................................................................................................166 
Listing 27: 16.2.8.2.1: Clinical Laboratory Results: Biochemistry - Asymptomatic 
Amebiasis Group ......................................................................................................167 
Listing 28: 16.2.8.2.2: Clinical Laboratory Results: Biochemistry - Asymptomatic 
Giardiasis Group .......................................................................................................167 
Listing 29: 16.2.8.3.1: Clinical Laboratory Results: Urinalysis - Asymptomatic 
Amebiasis Group ......................................................................................................168 
Listing 30: 16.2.8.3.2: Clinical Laboratory Results: Urinalysis - Asymptomatic 
Giardiasis Group .......................................................................................................168 
Listing 31: 16.2.9.1.1: Vital Signs - Asymptomatic Amebiasis Group ......................................169 
Listing 32: 16.2.9.1.2: Vital Signs - Asymptomatic Giardiasis Group .......................................169 
Listing 33: 16.2.9.2.1: Physical Exam Findings - Asymptomatic Amebiasis Group .................170 
Listing 34: 16.2.9.2.2: Physical Exam Findings - Asymptomatic Giardiasis Group ..................170 
Listing 35: 16.2.10.1: Concomitant Medications - Asymptomatic Amebiasis Group ................171 
Listing 36: 16.2.10.2: Concomitant Medications - Asymptomatic Giardiasis Group ................171 
Listing 37: 16.2.11.1.1: Pregnancy Reports - Maternal Information ..........................................172 
Listing 38: 16.2.11.1.2: Pregnancy Report - Gravida and Para ..................................................172 
Listing 39: 16.2.11.2.1: Pregnancy Reports – Live Birth Outcomes ..........................................173 
Listing 40: 16.2.11.2.2: Pregnancy Reports – Still Birth Outcomes ...........................................173 
Listing 41: 16.2.11.2.3: Pregnancy Reports - Spontaneous, Elective, or Therapeutic 
Abortion Outcomes ...................................................................................................173 
 


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16.1.6: Listing of Subjects Receiving Investigational Product 
(not included in SAP, but this is a placeholder for the CSR) 
 


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16.2 
Database Listings by Subject 
16.2.1 Discontinued Subjects 
Listing 1: 
16.2.1.1: Early Terminations or Discontinued Subjects - Asymptomatic Amebiasis Group 
Subject ID 
Treatment Group 
Category 
Reason for Early Termination or Treatment Discontinuation 
Study Day 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 2 : 
16.2.1.2: Early Terminations or Discontinued Subjects - Asymptomatic Giardiasis Group 
Subject ID 
Treatment Group 
Category 
Reason for Early Termination or Treatment Discontinuation 
Study Day 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


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16.2.2 Protocol Deviations 
Listing 3:  
16.2.2.1: Subject-Specific Protocol Deviations -Asymptomatic Amebiasis Group 
Subject 
ID 
Treatment 
Group 
DV Number 
Deviation 
Deviation 
Category 
Study Day 
Reason for 
Deviation 
Deviation 
Resulted 
in AE? 
Deviation Resulted 
in 
Subject 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
Listing 4: 
16.2.2.2: Subject-Specific Protocol Deviations - Asymptomatic Giardiasis Group 
Subject 
ID 
Treatment 
Group 
DV Number 
Deviation 
Deviation 
Category 
Study Day 
Reason for 
Deviation 
Deviation 
Resulted 
in AE? 
Deviation Resulted 
in 
Subject 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
 


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Listing 5: 
16.2.2.2.3: Non-Subject-Specific Protocol Deviations -Asymptomatic Amebiasis Group 
Site 
Deviation 
Start 
Date 
End Date 
Reason for 
Deviation 
Deviation 
Resulted in 
Subject 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Category 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
Listing 6:  
16.2.2.2.4: Non-Subject-Specific Protocol Deviations - Asymptomatic Giardiasis Group 
Site 
Deviation 
Start 
Date 
End Date 
Reason for 
Deviation 
Deviation 
Resulted in 
Subject 
Termination? 
Deviation 
Affected 
Product 
Stability? 
Deviation 
Category 
Deviation 
Resolution 
Comments 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
 


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16.2.3 Subjects Excluded from the Efficacy Analysis 
Listing 7:  
16.2.3.1: Subjects Excluded from Analysis Populations - Asymptomatic Amebiasis Group 
Treatment Group 
Subject ID 
Analyses in which 
Subject is Included 
Analyses from which  
Subject is Excluded 
Results Available? 
Reason Subject Excluded 
 
 
[e.g., Safety, m-ITT, PP] 
[e.g., Safety, m-ITT, PP] 
 
 
 
 
 
 
 
 
Note: “Yes” in the “Results available” column indicates that available data were removed from the analysis.  “No” indicates that no data were available for inclusion in the analysis. 
Listing will be repeated for the symptomatic study population 
Listing 8: 
16.2.3.2: Subjects Excluded from Analysis Populations - Asymptomatic Giardiasis Group 
Treatment Group 
Subject ID 
Analyses in which 
Subject is Included 
Analyses from which  
Subject is Excluded 
Results Available? 
Reason Subject Excluded 
 
 
[e.g., Safety, m-ITT, PP] 
[e.g., Safety, m-ITT, PP] 
 
 
 
 
 
 
 
 
Note: “Yes” in the “Results available” column indicates that available data were removed from the analysis.  “No” indicates that no data were available for inclusion in the analysis. 
Listing will be repeated for the symptomatic study population. 
 


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16.2.4 Demographic Data 
Listing 9:  
16.2.4.1: Demographic Data - Asymptomatic Amebiasis Group 
Subject ID 
Treatment Group 
Sex 
Age at Enrollment (years) 
Ethnicity 
Race 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
Listing 10:  
16.2.4.2: Demographic Data - Asymptomatic Giardiasis Group 
Subject ID 
Treatment Group 
Sex 
Age at Enrollment (years) 
Ethnicity 
Race 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
 


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Listing 11: 
16.2.4.3: Pre-Existing Medical Conditions - Asymptomatic Amebiasis Group 
Subject ID 
Treatment 
Group 
MH Number 
Medical History Term 
Condition Start Day  
Condition End Day 
MedDRA System Organ Class 
MedDRA Preferred Term 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
Listing 12: 
16.2.4.4: Pre-Existing Medical Conditions - Asymptomatic Giardiasis Group 
Subject ID 
Treatment 
Group 
MH Number 
Medical History Term 
Condition Start Day  
Condition End Day 
MedDRA System Organ Class 
MedDRA Preferred Term 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
 


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16.2.5 Compliance and/or Drug Concentration Data (if available) 
Listing 13: 
16.2.5.1: Compliance and/or Drug Concentration Data - Asymptomatic Amebiasis Group 
Subject ID 
Treatment Group 
Dose(s) Missed 
 
 
[e.g., Day 3, Day 5, etc.] 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
Listing 14: 
16.2.5.2: Compliance and/or Drug Concentration Data - Asymptomatic Giardiasis Group 
Subject ID 
Treatment Group 
Dose(s) Missed 
 
 
[e.g., Day 3, Day 5, etc.] 
 
 
 
 
 
 
 
 
 
Listing will be repeated for the symptomatic study population. 
 


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16.2.6 Individual Efficacy/Immunogenicity Response Data 
Listing 15: 
16.2.6.1: Listing of Efficacy Endpoints - Asymptomatic Amebiasis Group 
Treatment 
Group 
Subject ID 
Study 
Day 
Parasite Load 
(parasites/uL) 
Antigen Test 
Wet mount 
result 
Trichrome 
staining  
Parasitological Response 
(no detection of cysts or 
trophozoites or a negative 
antigen test) 
Relapse  (Strain 
genetically matches 
Day 1 strain)* 
Reinfection  (Strain 
genetically matches 
Day 1 strain)* 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Yes, no, or NA if no parasites are detected on the specified day. 
Listing 16: 
16.2.6.2: Listing of Efficacy Endpoints - Symptomatic Amebiasis Group 
Treatment 
Group 
Subject ID 
Study Day 
Diarrhea  
Parasite Load 
(parasites/uL) 
Antigen Test 
Wet mount 
Trichrome 
staining 
Parasitological 
Response (no 
detection 
trophozoites by 
microscopy) 
Relapse  (Strain 
genetically 
matches Day 1 
strain)* 
Reinfection  (Strain 
genetically matches 
Day 1 strain)* 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Yes, no, or NA if no parasites are detected on the specified day. 


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Listing 17: 
16.2.6.3: Listing of Efficacy Endpoints - Asymptomatic Giardiasis Group 
Treatment 
Group 
Subject ID 
Study 
Day 
Parasite Load 
(parasites/uL) 
Antigen Test 
Wet mount  
Trichrome 
staining  
Parasitological Response 
(no detection of cysts or 
trophozoites or a negative 
antigen test) 
Relapse  (Strain 
genetically matches 
Day 1 strain)* 
Reinfection  (Strain 
genetically matches 
Day 1 strain)* 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Yes, no, or NA if no parasites are detected on the specified day. 
Listing 18:  
16.2.6.4: Listing of Efficacy Endpoints - Symptomatic Giardiasis Group 
Treatment 
Group 
Subject ID 
Study Day 
Diarrhea  
Parasite Load 
(parasites/uL) 
Antigen Test 
Wet mount 
Trichrome 
staining 
Parasitological 
Response (no 
detection 
trophozoites by 
microscopy) 
Relapse  (Strain 
genetically 
matches Day 1 
strain)* 
Reinfection  (Strain 
genetically matches 
Day 1 strain)* 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
*Yes, no, or NA if no parasites are detected on the specified day 
 


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16.2.7 Adverse Events 
Listing 19: 
16.2.7.1.1: Solicited Events - Asymptomatic Amebiasis Group – First listing 
Subject ID 
Treatment 
Group 
Number of 
doses 
received 
Study 
Day 
Loose stools 
or diarrhea 
Abdominal Pain 
Nausea with 
or without 
vomiting 
Constipation 
Anorexia 
Flatulence 
Dyspepsia 
Dysgeusia 
 
 
 
 
[severity] 
[severity] 
[severity] 
[severity] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* MA = Data reported by subject on the Memory Aid and reviewed by clinic staff and reported in Solicited Events eCRF. 
Clinic = Data collected by clinic staff during physical exam or symptom assessment (treatment administration record, in-clinic assessment, etc.) 
Listing 20: 
16.2.7.1.2: Solicited Events - Asymptomatic Amebiasis Group – Second listing 
Subject ID 
Treatment 
Group 
Number of 
doses 
received 
Study 
Day 
Pruritus 
Hair loss 
Urticaria 
Stomatitis 
Conjunctivitis 
Glossitis 
Hematuria 
Rash 
 
 
 
 
[severity] 
[severity] 
[severity] 
[severity] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* MA = Data reported by subject on the Memory Aid and reviewed by clinic staff and reported in Solicited Events eCRF. 
Clinic = Data collected by clinic staff during physical exam or symptom assessment (treatment administration record, in-clinic assessment, etc.) 
Listing will be repeated for the symptomatic study population but will omit diarrhea. 
 


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Listing 21: 
16.2.7.1.2: Solicited Events - Asymptomatic Giardiasis Group – First listing 
Subject ID 
Treatment 
Group 
Number of 
doses 
received 
Study 
Day 
Loose stools 
or diarrhea 
Abdominal Pain 
Nausea with 
or without 
vomiting 
Constipation 
Anorexia 
Flatulence 
Dyspepsia 
Dysgeusia 
 
 
 
 
[severity] 
[severity] 
[severity] 
[severity] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* MA = Data reported by subject on the Memory Aid and reviewed by clinic staff and reported in Solicited Events eCRF. 
Clinic = Data collected by clinic staff during physical exam or symptom assessment (treatment administration record, in-clinic assessment, etc.) 
Listing 22: 
16.2.7.1.3: Solicited Events Asymptomatic Giardiasis Group – Second listing 
Subject ID 
Treatment 
Group 
Number of 
doses 
received 
Study 
Day 
Pruritus 
Hair loss 
Urticaria 
Stomatitis 
Conjunctivitis 
Glossitis 
Hematuria 
Rash 
 
 
 
 
[severity] 
[severity] 
[severity] 
[severity] 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* MA = Data reported by subject on the Memory Aid and reviewed by clinic staff and reported in Solicited Events eCRF. 
Clinic = Data collected by clinic staff during physical exam or symptom assessment (treatment administration record, in-clinic assessment, etc.) 
Listing will be repeated for the symptomatic study population but will omit diarrhea. 
 


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Listing 23: 
16.2.7.2.1: Unsolicited Adverse Events - Asymptomatic Amebiasis Group 
Adverse 
Event 
Associated 
with Dose 
No. 
No. of Days 
Post 
Associated 
Dose 
(Duration) 
Severity 
SAE? 
Relationship 
to Study 
Treatment 
In Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA 
System 
Organ Class 
MedDRA 
Preferred 
Term 
Treatment Group: , Subject ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment Group: , Subject ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
Note: For additional details about SAEs, see Table: xx. 
Listing will be repeated for the symptomatic study population. 
 
 


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Listing 24: 
16.2.7.2.2: Unsolicited Adverse Events - Asymptomatic Giardiasis Group 
Adverse 
Event 
Associated 
with Dose 
No. 
No. of Days 
Post 
Associated 
Dose 
(Duration) 
Severity 
SAE? 
Relationship 
to Study 
Treatment 
In Not 
Related, 
Alternative 
Etiology 
Action 
Taken with 
Study 
Treatment 
Subject 
Discontinued 
Due to AE 
Outcome 
MedDRA 
System 
Organ Class 
MedDRA 
Preferred 
Term 
Treatment Group: , Subject ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
 
 
 
 
 
 
 
 
 
 
 
 
Treatment Group: , Subject ID: , AE Number:  
 
 
 
 
 
 
 
 
 
 
 
 
Comments: 
Note: For additional details about SAEs, see Table: xx. 
Listing will be repeated for the symptomatic study population. 
 


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16.2.8 Individual Laboratory Measurements 
Listing 25: 
16.2.8.1.1: Clinical Laboratory Results: Hematology - Asymptomatic Amebiasis Group 
Subject ID 
Treatment 
Group 
Sex 
Age 
(years) 
Planned 
Time 
Point 
Actual 
Study 
Day 
White Blood Cells 
(109/L) 
Hemoglobin 
(g/dL) 
Hematocrit   
(% Decrease from 
baseline) 
Platelets  
(109/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 26: 
16.2.8.1.2: Clinical Laboratory Results: Hematology - Asymptomatic Giardiasis Group 
Subject ID 
Treatment 
Group 
Sex 
Age 
(years) 
Planned 
Time 
Point 
Actual 
Study 
Day 
White Blood Cells 
(109/L) 
Hemoglobin 
(g/dL) 
Hematocrit   
(% Decrease from 
baseline) 
Platelets  
(109/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listings will be repeated for the symptomatic study population. 
 


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Listing 27: 
16.2.8.2.1: Clinical Laboratory Results: Biochemistry - Asymptomatic Amebiasis Group 
Subject 
ID 
Treatment 
Group 
Sex 
Age 
(years) 
Planned 
Time 
Point 
Actual 
Study 
Day 
Blood Urea Nitrogen 
(mg/dL) 
Creatinine 
(mg/dL) 
Aspartate 
Aminotransferase 
(IU/L) 
Alanine 
Aminotransferase 
(IU/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 28: 
16.2.8.2.2: Clinical Laboratory Results: Biochemistry - Asymptomatic Giardiasis Group 
Subject 
ID 
Treatment 
Group 
Sex 
Age 
(years) 
Planned 
Time 
Point 
Actual 
Study 
Day 
Blood Urea Nitrogen 
(mg/dL) 
Creatinine 
(mg/dL) 
Aspartate 
Aminotransferase 
(IU/L) 
Alanine 
Aminotransferase 
(IU/L) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listings will be repeated for the symptomatic study population. 
 


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Listing 29: 
16.2.8.3.1: Clinical Laboratory Results: Urinalysis - Asymptomatic Amebiasis Group 
Subject ID 
Treatment Group 
Sex 
Age (years) 
Planned Time 
Point 
Actual Study 
Day 
Protein  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 30: 
16.2.8.3.2: Clinical Laboratory Results: Urinalysis - Asymptomatic Giardiasis Group 
Subject ID 
Treatment Group 
Sex 
Age (years) 
Planned Time 
Point 
Actual Study 
Day 
Protein  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listings will be repeated for the symptomatic study population. 
 


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16.2.9 Vital Signs and Physical Exam Findings 
Listing 31: 
16.2.9.1.1: Vital Signs - Asymptomatic Amebiasis Group 
Subject ID 
Treatment Group  
Planned Time 
Point 
Actual Study 
Day 
Temperature 
(°C) 
Systolic Blood 
Pressure 
(mmHg) 
Diastolic Blood 
Pressure 
(mmHg) 
Heart Rate 
(beats/min) 
Respiratory Rate 
(breaths/min) 
Weight 
(kg) 
Height 
(cm) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 32: 
16.2.9.1.2: Vital Signs - Asymptomatic Giardiasis Group 
Subject ID 
Treatment Group  
Planned Time 
Point 
Actual Study 
Day 
Temperature 
(°C) 
Systolic Blood 
Pressure 
(mmHg) 
Diastolic Blood 
Pressure 
(mmHg) 
Heart Rate 
(beats/min) 
Respiratory Rate 
(breaths/min) 
Weight 
(kg) 
Height 
(cm) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listings will be repeated for the symptomatic study population. 
 


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Listing 33: 
16.2.9.2.1: Physical Exam Findings - Asymptomatic Amebiasis Group 
Subject ID 
Treatment Group 
Planned Time 
Point 
Actual Study 
Day 
Body System 
Abnormal Finding 
Reported as an AE? 
(AE Number) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 34: 
16.2.9.2.2: Physical Exam Findings - Asymptomatic Giardiasis Group 
Subject ID 
Treatment Group 
Planned Time 
Point 
Actual Study 
Day 
Body System 
Abnormal Finding 
Reported as an AE? 
(AE Number) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listings will be repeated for the symptomatic study population. 
 


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16.2.10 Concomitant Medications 
Listing 35: 
16.2.10.1: Concomitant Medications - Asymptomatic Amebiasis Group 
Subject ID 
Treatment 
Group 
CM Number 
Medication 
Medication Start Day 
Medication End Day 
Indication 
Taken for an AE? 
(AE Number) 
Taken for a condition on 
Medical History? 
(MH Number) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 36: 
16.2.10.2: Concomitant Medications - Asymptomatic Giardiasis Group 
Subject ID 
Treatment 
Group 
CM Number 
Medication 
Medication Start Day 
Medication End Day 
Indication 
Taken for an AE? 
(AE Number) 
Taken for a condition on 
Medical History? 
(MH Number) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listings will be repeated for the symptomatic study population. 
 


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16.2.11.1 
Pregnancy Reports 
Listing 37: 
16.2.11.1.1: Pregnancy Reports - Maternal Information  
Subject 
ID 
Study 
population 
and disease 
group  
Treatment 
Group 
Pregnancy 
Number 
Study Day 
Corresponding 
to Estimated 
Date of 
Conception 
Source of 
Maternal 
Information 
Pregnancy 
Status 
Mother’s 
Pre-
Pregnancy 
BMI 
Mother’s 
Weight 
Gain 
During 
Pregnancy 
Tobacco, 
Alcohol, or 
Drug Use 
During 
Pregnancy? 
Medications 
During 
Pregnancy? 
Maternal 
Complications 
During 
Pregnancy? 
Maternal 
Complications 
During Labor, 
Delivery, or Post-
Partum? 
 
Symptomatic 
giardiasis 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: Maternal Complications are included in the Adverse Event listing.  Medications taken during pregnancy are included in the Concomitant Medications Listing. 
Listing 38: 
16.2.11.1.2: Pregnancy Report - Gravida and Para 
 
 
 
 
Live Births 
 
 
 
 
 
Subject 
ID 
Study 
population 
and 
disease 
group 
Pregnancy 
Number 
Gravida 
Extremely 
Preterm 
Births 
Very 
Preterm 
Births 
Early 
Preterm 
Births 
Late 
Preterm 
Births 
Early 
Term 
Births 
Full 
Term 
Births 
Late 
Term 
Births 
Post 
Term 
Births 
Still 
Births 
Spontaneous 
Abortion/ 
Miscarriage 
Elective 
Abortions 
Therapeutic 
Abortions 
Major 
Congenital 
Anomaly 
with 
Previous 
Pregnancy? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: Gravida includes the current pregnancy, para events do not. 
 
 


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16.2.11.2: Pregnancy Reports – Gravida and Para 
Listing 39: 
16.2.11.2.1: Pregnancy Reports – Live Birth Outcomes 
Subject 
ID 
Study 
population 
and disease 
group 
Pregnancy 
Number 
Fetus 
Number 
Pregnancy 
Outcome 
(for this 
Fetus) 
Fetal 
Distress 
During 
Labor 
and 
Delivery? 
Delivery 
Method 
Gestational 
Age at Live 
Birth  
Size for 
Gestational 
Age 
Apgar 
Score, 
1 
minute 
Apgar 
Score, 5 
minutes 
Cord 
pH 
Congenital 
Anomalies? 
Illnesses/ 
Hospitalizations 
within 1 Month of 
Birth? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Note: Congenital Anomalies are included in the Adverse Event listing. 
Listing 40: 
16.2.11.2.2: Pregnancy Reports – Still Birth Outcomes 
Subject 
ID 
Study 
population 
and disease 
group 
Date of 
Initial 
Report 
Fetus 
Number 
Pregnancy 
Outcome 
(for this 
Fetus) 
Fetal 
Distress 
During 
Labor and 
Delivery? 
Delivery 
Method 
Gestational 
Age at Still 
Birth 
Size for 
Gestational 
Age 
Cord 
pH 
Congenital 
Anomalies? 
Autopsy 
Performed? 
If Autopsy, 
Etiology for Still 
Birth Identified? 
 
 
 
 
 
 
 
 
 
 
 
 
 
Listing 41: 
16.2.11.2.3: Pregnancy Reports - Spontaneous, Elective, or Therapeutic Abortion Outcomes 
Subject 
ID 
Study population and 
disease group 
Date of Initial 
Report 
Fetus Number 
Pregnancy Outcome 
(for this Fetus) 
Gestational Age at 
Termination 
Abnormality in 
Product of 
Conception? 
Reason for Therapeutic 
Abortion