A Phase 1, Randomized, Double-blinded, Four-period Crossover, Thorough QT/QTC (TQT) Clinical Trial to Evaluate the Effect of Zoliflodacin on Cardiac Repolarization in Healthy Male and Female Subjects
Phase: PHASE1
Conditions: Electrocardiogram Repolarisation Abnormality, Gonococcal Infection
Sponsor: National Institute of Allergy and Infectious Diseases (NIAID)

 
 
A Phase 1, Randomized, Double-blinded, Four-period Crossover, 
Thorough QT/QTc (TQT) Clinical Trial to Evaluate the Effect of 
Zoliflodacin on Cardiac Repolarization in Healthy Male and Female 
Subjects 
 
 
DMID Protocol Number: 16-0110 
 
 
DMID Funding Mechanism: Phase 1 Clinical Trial Unit 
 
 
Pharmaceutical Support: Entasis Therapeutics 
 
 
IND Sponsor: Division of Microbiology and Infectious Diseases (DMID), 
National Institute of Allergy and Infectious Diseases (NIAID), National Institutes 
of Health (NIH) 
 
 
Site Principal Investigator: Martin Kankam, MD, PhD, MPH 
 
 
DMID Clinical Project Manager: Meg Gordon, RN 
 
 
Version Number: 4.0 
 
 
27 November 2018 
 
 

DMID Protocol 16-0110 
Version 4.0 
Zoliflodacin 
27 November 2018 
 ______________________________________________________________________________________________________  
DMID/NIAID/NIH 
CONFIDENTIAL 
 
 
 
2 
Statement of Compliance 
The clinical trial will be carried out in accordance with Good Clinical Practice (GCP) and as 
required by the following: 
• United States (US) Code of Federal Regulations (CFR) applicable to clinical trials (45 CFR 
Part 46, 21 CFR Part 50, 21 CFR Part 54, 21 CFR Part 56, and 21 CFR Part 312) 
• International Council for Harmonization of Technical Requirements for Registration of 
Pharmaceuticals for Human Use E6(R2) Good Clinical Practice (ICH E6 GCP): Integrated 
Addendum to ICH E6(R1) Guidance for Industry, published in the Federal Register (83 
Federal Register 8882 (2018)), including the latest finalized.National Institutes of Health 
(NIH) Clinical Terms of Award, as applicable 
Compliance with these standards provides public assurance that the rights, safety, and well-being 
of subjects are protected, consistent with principles that originate in the Declaration of Helsinki. 
All key personnel (all individuals responsible for the design and conduct of the trial) have 
completed Human Subjects Protection Training.  
 
 

DMID Protocol 16-0110 
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Zoliflodacin 
27 November 2018 
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DMID/NIAID/NIH 
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3 
Signature Page 
The signature below constitutes the approval of the protocol and its attachments and provides the 
necessary assurances that the 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 US federal regulations and ICH guidelines. 
 
 
 
Site Investigator: 
 
 
_________________________________________________ 
Name/Title (Print) 
 
 
Signature: 
 
_________________________________________________ 
 
Date:  
 
_________________________________________________ 
 
 
 
 
 
 

DMID Protocol 16-0110 
Version 4.0 
Zoliflodacin 
27 November 2018 
 ______________________________________________________________________________________________________  
DMID/NIAID/NIH 
CONFIDENTIAL 
 
 
 
4 
Table of Contents 
 
 
Page 
Statement of Compliance .................................................................................................................2 
Signature Page .................................................................................................................................3 
Table of Contents .............................................................................................................................4 
List of Tables ...................................................................................................................................9 
List of Figures ................................................................................................................................10 
List of Abbreviations .....................................................................................................................11 
Protocol Summary .........................................................................................................................15 
1 
Key Roles ..............................................................................................................................19 
2 
Background Information and Scientific Rationale ...............................................................21 
2.1 
Background Information ...........................................................................................21 
2.1.1 Overview of the Disease: Gonorrhea and its Treatment ...............................21 
2.1.2 Zoliflodacin Development ............................................................................21 
2.2 
Rationale ...................................................................................................................26 
2.2.1 Study Rationale .............................................................................................26 
2.2.2 Rationale for Dose Selection ........................................................................27 
2.3 
Identified and Potential Risks and Benefits ..............................................................28 
2.3.1 Zoliflodacin Risks .........................................................................................28 
2.3.2 Moxifloxacin Risks .......................................................................................30 
2.3.3 Venipuncture and IV Catheter Placement Risks ...........................................31 
2.3.4 Additional Risks............................................................................................31 
2.3.5 Known Potential Benefits .............................................................................32 
2.3.6 Risk/Benefit Ratio .........................................................................................32 
3 
Objectives and Outcome Measures.......................................................................................33 
3.1 
Study Objectives .......................................................................................................33 
3.1.1 Primary ..........................................................................................................33 
3.1.2 Secondary ......................................................................................................33 
3.2 
Study Outcome Measures .........................................................................................33 
3.2.1 Primary ..........................................................................................................33 
3.2.2 Secondary ......................................................................................................33 
4 
Study Design .........................................................................................................................35 
4.1 
Sub-studies ................................................................................................................36 
5 
Study Enrollment and Withdrawal .......................................................................................37 
5.1 
Subject Inclusion Criteria .........................................................................................37 
5.2 
Subject Exclusion Criteria ........................................................................................38 
5.3 
Treatment Assignment Procedures ...........................................................................41 
5.3.1 Enrollment and Randomization Procedures..................................................41 
5.3.2 Masking Procedures ......................................................................................42 

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27 November 2018 
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DMID/NIAID/NIH 
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5.3.3 Reasons for Withdrawal and Discontinuation of Study Drug 
Administration ..............................................................................................42 
5.3.4 Handling of Withdrawals and Discontinuation of Administration ...............43 
5.3.5 Lost to Follow-up ..........................................................................................43 
5.3.6 Termination of Study ....................................................................................43 
6 
Study Intervention/investigational product ...........................................................................44 
6.1 
Description of Study Products ..................................................................................44 
6.1.1 Acquisition ....................................................................................................44 
6.1.2 Formulation, Packaging, and Labeling .........................................................45 
6.1.3 Product Storage and Stability........................................................................45 
6.2 
Dosage, Preparation, and Administration of Study Intervention/ Investigational 
Products.....................................................................................................................46 
6.3 
Modification of the Study Intervention/ Investigational Products for a Subject ......47 
6.3.1 Overdose .......................................................................................................47 
6.4 
Accountability Procedures for the Study Intervention/ Investigational Products .....47 
6.5 
Assessment of Subject Compliance with the Study Intervention/ Investigational 
Products.....................................................................................................................48 
6.6 
Prior and  Concomitant Medications/Treatments .....................................................48 
6.7 
Subject Restrictions ..................................................................................................49 
6.7.1 Physical Activity ...........................................................................................49 
6.7.2 Fluid and Food intake ...................................................................................49 
6.7.3 Alcohol, Marijuana and Illicit Drugs ............................................................50 
6.7.4 Other Restrictions .........................................................................................50 
7 
Study Schedule .....................................................................................................................51 
7.1 
Recruitment ...............................................................................................................51 
7.2 
Screening Visit (Day -21 to Day -2) .........................................................................51 
7.3 
Check-in / Baseline (Day-1 in each dosing period)                                          
(Overall, Days -1, 8, 16, and 24 for dosing periods 1, 2, 3 and 4, respectively) ......52 
7.4 
Inpatient period (Days 1-2 in each dosing period)                                          
(Overall, Days 1-2, 9-10, 17-18, and 25-26 for dosing periods 1, 2, 3 and 4,  
respectively) ..............................................................................................................53 
7.4.1 Randomization (Day 1, Dosing Period 1 only) .............................................53 
7.4.2 Administration of Study Drug (Day 1 of each dosing period)                          
(Overall, Days 1, 9, 17, and 25 for dosing periods 1, 2, 3 and 4, 
respectively) ..................................................................................................53 
7.4.3 Inpatient Follow-up (Day 2 of each dosing period)                                            
(Overall, Days 2, 10, 18, and 26 for dosing periods 1, 2 3 and 4, 
respectively) ..................................................................................................55 

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7.5 
Outpatient period (Days 3-7 for each dosing period                           (Overall, 
Days 3-7, 11-15, 19-23 and 27-31 for dosing periods 1, 2, 3 and 4, respectively) ...55 
7.6 
Final Visit (Day 32, 8 ±2 days after last dose in Dosing Period 4) ..........................55 
7.7 
Early Termination Visit (if needed) ..........................................................................56 
7.8 
Unscheduled Visit (if needed) ..................................................................................56 
8 
Study Procedures/evaluations ...............................................................................................57 
8.1 
Clinical Procedures/Evaluations ...............................................................................57 
8.1.1 Informed Consent..........................................................................................57 
8.1.2 Demographics ...............................................................................................57 
8.1.3 Inclusion/Exclusion Criteria .........................................................................57 
8.1.4 Medical History ............................................................................................57 
8.1.5 Physical Examination....................................................................................58 
8.1.6 Vital Signs .....................................................................................................58 
8.1.7 12-lead Standard Electrocardiogram (ECG) .................................................59 
8.2 
Laboratory Evaluations .............................................................................................59 
8.2.1 Clinical Laboratory Evaluations (Hematology, Chemistry, and 
Urinalysis) .....................................................................................................59 
8.2.2 Viral Serology Testing ..................................................................................60 
8.2.3 Pregnancy Testing .........................................................................................60 
8.2.4 Serum FSH Testing .......................................................................................60 
8.2.5 Urine Toxicology Screening .........................................................................60 
8.2.6 Urine Alcohol Testing...................................................................................61 
8.3 
Special Procedures and Assays .................................................................................61 
8.3.1 Continuous 24-h ECG Monitoring (Holter ECG Monitoring) .....................61 
8.3.1.1 TQT and ECG Extraction Technique ...............................................62 
8.3.1.2 T-wave Morphology Analysis and Measurement of PR and QRS 
Intervals.............................................................................................62 
8.3.2 Pharmacokinetics ..........................................................................................63 
8.3.2.1 Assay for Zoliflodacin ......................................................................63 
8.3.2.2 Assays for Placebo and Moxifloxacin Samples ................................63 
8.3.3 Specimen Preparation, Handling, and Shipping ...........................................64 
8.3.3.1 Instructions for Specimens Preparation, Handling, and Storage ......64 
8.3.3.2 Specimen Shipment ..........................................................................64 
9 
Assessment of Safety ............................................................................................................65 
9.1 
Specification of Safety Parameters ...........................................................................65 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters .................................................................................................................65 
9.2.1 Adverse Events .............................................................................................65 
9.2.1.1 Severity of Events .............................................................................66 

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9.2.1.1.1 Relationship to Study Products ........................................... 66 
9.2.1.2 Reporting Adverse Events ................................................................66 
9.2.2 Serious Adverse Events ................................................................................67 
9.2.3 Procedures to be Followed in the Event of Abnormal Laboratory Test 
Values or Abnormal Clinical Findings .........................................................67 
9.3 
Reporting Procedures ................................................................................................68 
9.3.1 Serious Adverse Events ................................................................................68 
9.3.2 Regulatory Reporting for Studies Conducted Under DMID-Sponsored 
IND ...............................................................................................................69 
9.3.3 Other Adverse Events (if applicable) ............................................................69 
9.3.3.1 Reporting of Overdose ......................................................................69 
9.3.4 Reporting of Pregnancy ................................................................................70 
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events .........................70 
9.5 
Halting Rules ............................................................................................................70 
9.5.1 Study Halting Criteria ...................................................................................70 
9.5.1.1 Halting Criteria for Zoliflodacin and Placebo Periods .....................70 
9.5.1.2 Halting Criteria for Moxifloxacin Periods ........................................71 
9.6 
Safety Oversight........................................................................................................71 
9.6.1 Independent Safety Monitor (ISM) ...............................................................71 
9.6.2 Safety Monitoring Committee (SMC) ..........................................................71 
10 
Clinical Monitoring...............................................................................................................73 
10.1 
Site Monitoring Plan .................................................................................................73 
11 
Statistical Considerations ......................................................................................................74 
11.1 
Study Hypotheses......................................................................................................74 
11.2 
Sample Size Considerations ......................................................................................74 
11.3 
Safety Review ...........................................................................................................74 
11.4 
Final Analysis Plan ...................................................................................................74 
11.4.1 Analysis Populations .....................................................................................75 
11.4.2 Demographics and Baseline Characteristics .................................................75 
11.4.3 Safety Analysis .............................................................................................76 
11.4.3.1 Adverse and Serious Adverse Events ...............................................76 
11.4.3.2 Additional Safety Analyses...............................................................76 
11.4.4 PK Analysis ..................................................................................................76 
11.4.5 Missing Values and Outliers .........................................................................77 
11.4.6 Analysis of Continuous ECG Parameters (Pharmacodynamics) ..................77 
11.4.6.1 ECG Analysis and Interpretation ......................................................77 
11.4.6.2 Primary Analysis: QTcF Effect ........................................................77 
11.4.6.3 Secondary Analyses: Non-QTc Intervals..........................................78 
11.4.6.4 Assay Sensitivity ...............................................................................78 

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11.4.6.5 Categorical Analyses ........................................................................78 
11.4.6.6 Cardiac Abnormalities: .....................................................................79 
11.4.7 Concentration-QTcF Modeling .....................................................................79 
12 
Source Documents and Access to Source Data/Documents .................................................80 
13 
Quality Control and Quality Assurance ................................................................................81 
14 
Ethics/Protection of Human Subjects ...................................................................................82 
14.1 
Ethical Standard ........................................................................................................82 
14.2 
Institutional Review Board .......................................................................................82 
14.3 
Informed Consent Process ........................................................................................82 
14.4 
Exclusion of Women, Minorities, and Children (Special Populations) ....................84 
14.5 
Subject Confidentiality .............................................................................................84 
14.6 
Certificate of Confidentiality ....................................................................................84 
14.7 
Study Discontinuation ...............................................................................................85 
15 
Data Handling and Record Keeping .....................................................................................86 
15.1 
Data Management Responsibilities...........................................................................86 
15.2 
Data Capture Methods ..............................................................................................86 
15.3 
Types of Data ............................................................................................................87 
15.4 
Timing/Reports .........................................................................................................87 
15.5 
Study Records Retention...........................................................................................87 
15.6 
Protocol Deviations ...................................................................................................87 
16 
Publication Policy .................................................................................................................89 
17 
Literature References ............................................................................................................90 
18 
Appendices............................................................................................................................93 
Appendix A: Schedule of Study Procedures and Evaluations ...................................................... 94 
Appendix B: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements .......... 97 
Appendix C: Adverse Events Toxicity Grading Criteria ............................................................ 101 
Appendix D: Blood Volume Withdrawn During the Trial ......................................................... 109 
Appendix E: A Partial List of Drugs Causing QTc Prolongation and Torsade de Pointes ........ 111 
 
 

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List of Tables 
Table 1: Dosing ............................................................................................................................. 35 
Table 2: T-wave Morphology Categories ..................................................................................... 63 
Table 3: Schedule of Events ......................................................................................................... 94 
Table 4: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements – 
Hematology and Chemistry .......................................................................................................... 97 
Table 5: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements – Urinalysis
....................................................................................................................................................... 99 
Table 6: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements – Vital Signs
..................................................................................................................................................... 100 
Table 7: Toxicity Grading Tables – Clinical AEs ...................................................................... 102 
Table 8: Toxicity Grading Tables – Laboratory AEs ................................................................. 106 
Table 9: Toxicity Grading Tables – ECG ................................................................................... 108 
Table 10: Laboratory Samples and Estimated Total Blood Volume (mL) ................................. 109 
Table 11: A Partial List of Drugs Causing QTc Prolongation and Torsade de Pointes.............. 111 
 

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Zoliflodacin 
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10 
List of Figures 
Figure 1: Description Schematic of Study Design ........................................................................ 18 
 
 
 

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List of Abbreviations 
AE 
Adverse Event 
ALT 
Alanine Aminotransferase 
AP 
Alkaline Phosphatase 
AST 
Aspartate Aminotransferase 
AUC 
Area Under the Curve 
β-HCG 
Beta Human Chorionic Gonadotropin 
BID 
Twice-Daily 
BMI 
Body Mass Index 
BP 
Blood Pressure 
bpm 
Beats per Minute 
BUN 
Blood Urea Nitrogen 
CBC 
Complete Blood Count 
CDC 
Centers for Disease Control and Prevention 
CFR 
Code of Federal Regulations 
CHEM 
Chemistry Panel 
CI 
Confidence Interval 
Cmax 
Maximum Plasma Concentration 
CMS 
Clinical Materials Services 
ConMed(s) 
Concomitant Medication(s) 
CPM 
Clinical Project Manager 
CROMS 
Clinical Research Operations and Management Support 
CSR 
Clinical Study Report 
CTU 
Clinical Trial Unit 
DCC 
Data Coordinating Center 
ΔΔQTcF 
Time-matched, placebo-corrected, baseline-adjusted difference in QTcF 
interval 
dL 
Deciliter 
DMID 
Division of Microbiology and Infectious Diseases 
DNA 
Deoxyribonucleic Acid 
eCRF 
Electronic Case Report Form 

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12 
EC 
Ethics Committee 
ECG  
Electrocardiogram 
EDC 
Electronic Data Capture 
ET 
Early Termination 
FDA 
Food and Drug Administration 
FSH 
Follicle-Stimulating Hormone 
FWA 
Federalwide Assurance 
g 
Gram(s) 
GCP 
Good Clinical Practice 
GFR 
Glomerular Filtration Rate 
h 
Hour(s) 
HBsAg 
Hepatitis B Surface Antigen 
Hct 
Hematocrit 
HCV 
Hepatitis C Virus 
HEENT 
Head, Eyes, Ears, Nose, and Throat 
HEM 
Hematology Tests 
Hgb 
Hemoglobin 
HIV 
Human Immunodeficiency Virus 
HPF 
High-Powered Field 
HPLC-MS/MS 
High-Performance Liquid Chromatography with Tandem Mass Spectrometry 
 
HR 
Heart Rate 
IB 
Investigator’s Brochure 
IC50 
50% Inhibitory Concentration 
ICF 
Informed Consent Form 
ICH 
International Council for   Harmonization 
ICMJE 
International Committee of Medical Journal Editors 
IM 
Intramuscular 
IND 
Investigational New Drug  
IRB 
Institutional Review Board 
ISM 
Independent Safety Monitor 
IV 
Intravenous 

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13 
kg 
Kilogram(s) 
LLN 
Lower Limit of Normal 
LQTS 
Long QT Syndrome 
MDMA 
3,4-methylenedioxy-methamphetamine 
MedDRA 
Medical Dictionary for Regulatory Activities 
µg 
Microgram(s) 
mg 
Milligram(s) 
MH 
Medical History 
MIC 
Minimum Inhibitory Concentration 
min 
Minute(s) 
mL 
Milliliter(s) 
MM 
Medical Monitor 
mmHg 
Millimeters of Mercury 
µmol 
Micromole(s) 
MOP 
Manual of Procedures 
msec 
Milliseconds(s) 
mV 
Millivolt 
N 
Number (typically refers to the total number of subjects) 
n 
Number (typically refers to a subset of the total number of subjects) 
NIAID 
National Institute of Allergy and Infectious Diseases 
NIH 
National Institutes of Health 
NOAEL 
No Observable Adverse Effect Level 
NSAID 
Nonsteroidal Anti-Inflammatory Drug 
OCRR 
Office of Clinical Research Resources 
OTC 
Over-the-Counter 
PBPK 
Physiologically-Based Pharmacokinetic 
PD 
Pharmacodynamic(s) 
PE 
Physical Examination 
PI 
Principal Investigator 
PK 
Pharmacokinetic(s) 
PO 
Oral 

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POC 
Point of Contact 
PVG 
Pharmacovigilance Group 
QTc 
Corrected QT Interval of the ECG 
QTcF  
Corrected QT interval of the ECG using Fridericia’s Formula 
RBC 
Red Blood Cell 
RP 
Research Pharmacist 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SD 
Standard Deviation 
sec 
Second(s) 
SMC 
Safety Monitoring Committee 
TCA 
Tricyclic Antidepressant 
TdP 
Torsade de Pointes 
Tmax 
Time to Maximum Plasma Concentration 
TQT 
Thorough QT/QTc 
UA 
Urinalysis 
ULN 
Upper Limit of Normal 
US 
United States 
VS 
Vital Signs 
WBC 
White Blood Cell 
WHO 
World Health Organization 
 
 
 

DMID Protocol 16-0110 
Version 4.0 
Zoliflodacin 
27 November 2018 
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CONFIDENTIAL 
 
 
 
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Protocol Summary 
Title: 
A Phase 1, Randomized, Double-blinded, Four-period Crossover, Thorough 
QT/QTc (TQT) Clinical Trial to Evaluate the Effect of Zoliflodacin on Cardiac 
Repolarization in Healthy Male and Female Subjects 
Phase: 
1 
Population: 
72 randomized (at least 56 evaluable) healthy male and female subjects aged 
18 to 45 years inclusive  
Number of Sites: 
 
One (Vince & Associates Clinical Research, Inc., Overland Park, KS) 
Study Duration:  
 
Approximately 12 weeks  
Subject 
Participation 
Duration: 
 
Up to 55 days (from Screening Visit to Final Visit) 
The trial will consist of: 
• A Screening Period of up to 20 days (Day -21 to Day -2);  
• Four Dosing Periods, each lasting 3 days/2 nights (Days -1, 1, and 2 in each 
period), totaling 12 days;  
• Four Outpatient Periods, each lasting 5 days (Days 3-7 after each Dosing 
Period), totaling 20 days; and 
• A Final Visit 8 ±2 days after dosing in Dosing Period 4, totaling 1 day. 
Description of 
Agents: 
The following four treatments will be administered to all subjects using a 
randomized, double-blinded, four-period, crossover design. Each subject will 
be randomized to a treatment sequence containing all four treatments:  
• Treatment A: zoliflodacin 2 g (investigational product) 
• Treatment B: zoliflodacin 4 g (investigational product) 
• Treatment C: placebo (for zoliflodacin) 
• Treatment D: moxifloxacin 400 mg (positive comparator) 
 
Single 2-g and single 4-g doses of zoliflodacin were safe and well-tolerated in 
previous clinical trials. The 2-g dose (of the new formulation) has been 
selected as the therapeutic dose for further clinical testing of zoliflodacin. The 
4-g dose was shown to result in drug exposures higher than the 2-g dose and 
should provide exposures that will be achieved in patients when used with 
other drugs that may interact with zoliflodacin. Based on pharmacokinetic 

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(PK) data from study DMID 16-0118, exposures for 4 g of the new 
formulation are similar to those for the previous formulation. For this reason, 
the 4-g dose was selected as the supratherapeutic dose for this TQT trial. 
Placebo for zoliflodacin was selected to control for natural diurnal changes in 
the electrocardiogram (ECG). Moxifloxacin was selected as a positive 
comparator to evaluate QTcF assay sensitivity, as it predictably causes a mild 
prolongation of the QTcF interval but has a very low risk for inducing cardiac 
arrhythmias.   
Objectives: 
 
Primary: 
• To evaluate the effect of zoliflodacin on the corrected QT interval of 
the ECG using Fridericia’s formula (QTcF) 
Secondary:  
• To evaluate the effects of zoliflodacin on other ECG parameters [PR, 
QRS, and RR intervals, and heart rate (HR)]  
• To evaluate the sensitivity of QTcF measurement using moxifloxacin 
• To evaluate the effect of zoliflodacin on T-wave morphology 
• To evaluate the PK of 2 g and 4 g oral zoliflodacin under fasting state 
• To evaluate the relationship between zoliflodacin PK and time-matched 
QTcF pharmacodynamics (PD)  
• To evaluate the safety and tolerability of 2 g and 4 g oral zoliflodacin  
Outcome 
Measures:  
 
 
 
 
 
 
 
 
 
Primary: 
• The upper bound of the one-sided 95% confidence interval (CI) for the 
largest time-matched, placebo-corrected, baseline-adjusted mean QTcF 
interval (∆∆QTcF) collected in the 24-h period after 2 g and 4 g dose of 
zoliflodacin is <10 msec 
Secondary: 
• Time-matched, placebo-corrected, baseline-adjusted non-QT intervals 
(PR, QRS, and RR intervals) and HR collected in the 24-h period after 
2 g and 4 g dose of zoliflodacin  
• The lower bound of the one-sided 95% CI of the ∆∆QTcF is >5 msec 
at one or more individual time points between 1 h and 4 h after a single 
dose of moxifloxacin 400 mg  
• Incidence of abnormal T-wave morphology using defined categories in 
the 24-h period after 2 g and 4 g dose of zoliflodacin 

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• Single-dose plasma concentrations (and PK exposure parameters, such 
as Cmax and AUC) of zoliflodacin collected up to 24 h after 2 g and 4 g 
dose of zoliflodacin 
• Relationship between plasma concentrations of zoliflodacin (and PK 
exposure parameters, such as Cmax and AUC) and ∆∆QTcF up to 24 h 
after 2 g and 4 g dose of zoliflodacin 
• Occurrence of treatment-emergent serious adverse events (SAEs), other 
unsolicited treatment-emergent adverse events (AEs), and changes 
from baseline in vital signs (VS), clinical laboratory values, and ECG 
parameters following administration of zoliflodacin and moxifloxacin 
from time of dosing to Final Visit, or Early Termination (ET) Visit. 
Description of 
Study Design: 
The TQT clinical trial will be performed according to a randomized, double-
blinded (except for the use of moxifloxacin), placebo-controlled, four-period, 
four-treatment, crossover design in 72 healthy male and/or female subjects. 
Subjects who consent to participate will be enrolled in the trial if they meet all 
inclusion and no exclusion eligibility criteria. Laboratory assessments at 
Screening Visit will include clinical laboratory tests [hematology (HEM), 
chemistry (CHEM), and urinalysis (UA)], testing for viral serology [human 
immunodeficiency virus (HIV) antibody, hepatitis B surface antigen (HBsAg), 
and hepatitis C virus (HCV) antibody], drugs of abuse, and alcohol. Serum 
pregnancy testing will be done in all females and serum follicle-stimulating 
hormone (FSH) levels will be measured only in post-menopausal females. 
Subjects who meet eligibility criteria will be randomly allocated to one of 12 
dosing sequences and receive a single dose of the four study drugs according 
to their randomly-assigned dosing sequence during four dosing periods. The 
dosing interval between dosing periods will be 8 days to allow for adequate 
wash-out of drug before the next dose. Safety assessments at scheduled visits 
after dosing will include medical history (MH), VS, physical examination 
(PE), 12-lead ECG, and clinical laboratory tests. Continuous 12-lead Holter 
ECGs will be initiated 1 h before dosing and continue for 24 h after dosing. At 
scheduled timepoints, replicate Holter ECGs will be selected for measurement 
of QTcF and other ECG intervals, and assessment of T-wave morphologies. 
Plasma will be collected during each dosing period at scheduled timepoints 
that are time-matched to Holter ECGs before and after dosing on Day 1 and 
through Day 2 (24 h after dosing), or ET if within 24 h after dosing, but drug 
concentration bioanalysis and PK analysis will be done only for zoliflodacin. 
Treatment-emergent AEs and SAEs will be assessed from time of dosing to 
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Estimated Time 
to Complete 
Enrollment: 
Up to 12 weeks (from dosing of first subject to Final Visit of last subject) 
Halting Criteria 
Decision Process: 
If criteria for halting the trial (as listed in Section 9.5.1) are met following 
dosing in any dosing period, administration of all study drugs will be 
suspended and an ad hoc Safety Monitoring Committee (SMC) meeting will 
be held to review all available safety data and to make recommendations about 
the dosing of all further subjects in the trial. A scheduled SMC meeting will be 
held after all subjects complete the trial to review all safety data and advise on 
safety assessments in future clinical trials with zoliflodacin. 
 
 
Figure 1: Description Schematic of Study Design 
 
Screen
Washout between dosing
At least 8 days
Treatment
 Period 1
A
B
etc.
FINAL VISIT
8 + 2 days
After last dose
SCREENING
Up to 21 days
Before 1st dose
Treatment
 Period 2
B
C
etc.
Treatment
 Period 3
C
D
etc.
Treatment
 Period 4
D
A
etc.
Final Visit
Randomization 
Sequences*
ABCD
BCDA
etc.
RANDOMIZATION 
before dosing on
Day +1 of 
Treatment Period 1
 
 
NOTE: A, B, C, and D refer to each of the study treatments. This is a four-treatment, four-period, crossover trial. 
Treatments and treatment sequences will be randomized using a 12-sequence design balanced for period and first-
order carryover effects, and double-blinded, except for administration of moxifloxacin. Six subjects will be 
randomized to each treatment sequence. The schematic displays only two of the 12 sequences.  
* See Section 4 for details and a complete list of planned treatment sequences. 
 
 

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1 
KEY ROLES 
Site Principal Investigator: 
Martin Kankam, MD, PhD, MPH  
Vince & Associates Clinical Research, Inc. 
10103 Metcalf Avenue 
Overland Park, KS 66212 
Office: 913-696-1601   
Email: mkankam@vinceandassociates.com  
  
DMID Clinical Project Manager: 
Meg Gordon, RN 
DMID/NIAID/NIH 
5601 Fishers Lane, Room 7G37  
Bethesda, MD 20892-9826 
Office: 301-761-7604 
Email: meg.gordon@nih.gov  
 
Data Coordinating Center: 
The Emmes Corporation 
401 N. Washington Street, Suite 700 
Rockville, MD 20850 
Phone: 301-251-1161 
Fax: 301-576-3558 
Email: phase1@emmes.com  
 
ECG and Cardiac Services  
Quintiles Research (India) Private Ltd.  
Natraj by Rustomjee 
602 A, 6th Floor,  
194, MV Road,  
Near Western Express Highway Metro Station 
Andheri (East) 
Mumbai 400 069 
Maharashtra 
India 
Phone: +91 226 218 7000  
Email: Priti.Somani@quintiles.com 
 

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Bioanalytical Laboratory: 
KCAS Bioanalytical Services 
12400 Shawnee Mission Parkway 
Shawnee, KS 66216 
POC: Marsha Luna, Senior Manager, PBI 
Phone: 913-248-3042 
Email: marsha.luna@kcasbio.com 
  

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2 
BACKGROUND INFORMATION AND SCIENTIFIC 
RATIONALE 
2.1 
Background Information 
2.1.1 
Overview of the Disease: Gonorrhea and its Treatment 
Neisseria gonorrhoeae, currently the second most common bacterial sexually transmitted 
infection worldwide (after Chlamydia trachomatis), is a serious public health problem.1,2 In 
2008, the World Health Organization (WHO) estimated 106 million cases of gonorrhea among 
adults worldwide.3 In 2012, the Centers for Disease Control and Prevention (CDC) reported 
311,404 cases of gonorrhea in the US; however, due to incomplete reporting, up to 820,000 cases 
may occur annually.4 N. gonorrhoeae has developed resistance to all antimicrobial treatments for 
gonorrhea. The CDC recently released its report “Antibiotic Resistance Threats in the US, 2013,” 
which ranks drug-resistant N. gonorrhoeae as an “Urgent Threat,” defined as “an immediate 
public health threat that requires urgent and aggressive action.”5 This level of resistance has 
demanded alternative oral treatments, which until recently have been readily available.6 
Examples of drug classes that are widely used to treat gonorrhea, but are no longer recommended 
as monotherapy due to resistance, include sulfanilamides, penicillin, tetracyclines, and 
fluoroquinolones.1,2 Most recently, N. gonorrhoeae resistance to macrolides (including 
azithromycin), cefixime, and ceftriaxone, and consequent clinical failures, have been reported. 
Since two extended-spectrum cephalosporins, ceftriaxone and cefixime, have recently been the 
only first-line options for treating gonorrhea, resistance and treatment failures with these drugs 
are particularly concerning. It is only a matter of time before gonococci with full resistance to 
extended-spectrum cephalosporins emerge and spread globally.7 The threat of widespread 
ceftriaxone resistance and untreatable N. gonorrhoeae infection is real.5,6  The clinical 
development of zoliflodacin (previously called AZD0914 and ETX0914) addresses the need for 
new antibiotics that can be used alone or with other agents to treat uncomplicated gonorrhea 
caused by emergent N. gonorrhoeae strains that are resistant to existing antibiotics.8,9 
2.1.2 
Zoliflodacin Development  
Zoliflodacin is a spiropyrimidinetrione antibacterial drug, which inhibits bacterial DNA 
synthesis by a novel mechanism. Zoliflodacin demonstrates in vitro activity against 
N. gonorrhoeae strains that are susceptible or resistant to current available therapies and in vivo 
efficacy, including in patients with uncomplicated gonorrhea.8,9,10,11 
This section provides a summary of zoliflodacin. Detailed nonclinical and clinical data are 
provided in the Investigator’s Brochure (IB). 
 
 

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Chemistry 
The chemical name for zoliflodacin is (2R,4S,4aS)-11-fluoro-2,4-dimethyl-8-[(4S)-4-methyl-2-
oxo-1,3-oxazolidin-3-yl]-1,2,4,4a-tetrahydro-2'H,6H-spiro[1,4-oxazino[4,3-a][1,2]oxazolo[4,5-
g] quinoline-5,5'-pyrimidine]-2',4',6'(1'H,3'H)-trione. 
Mechanism of Action 
Zoliflodacin inhibits bacterial type II topoisomerases, specifically bacterial gyrases. The activity 
and mode of inhibition of bacterial gyrase by zoliflodacin has been assessed in several in vitro 
assays. Zoliflodacin stabilizes the 
 
 but in addition prevents 
 This novel 
mode of inhibition is further supported by the observation that
 
N. gonorrhoeae 
resistance to zoliflodacin has been mapped to 
.8 
Microbiological Activity  
In vitro: Zoliflodacin has demonstrated activity in vitro against N. gonorrhoeae, including strains 
that are resistant to penicillin, ciprofloxacin, tetracycline, azithromycin, ceftriaxone, and 
cefixime. Minimum inhibitory concentration (MIC)90 values ranged from 0.125 to 0.25 μg/mL. 
Zoliflodacin has also shown bactericidal activity at an MIC90 of 0.25 μg/mL against C. 
trachomatis, which often co-infects patients with gonorrhea.8,9,10  
In vivo: Mouse model of Staphylococcus aureus infection: An animal model for N. gonorrhoeae 
has not been established due to spontaneous bacterial eradication. To estimate a human 
efficacious exposure of zoliflodacin for N. gonorrhoeae, a surrogate pathogen approach, which 
utilizes PK/PD determinations from a S. aureus neutropenic mouse thigh model that correlates 
with human clinical efficacy, was used. In this model, zoliflodacin was efficacious in vivo 
against clinical S. aureus isolates. The MIC values of zoliflodacin against S. aureus and N. 
gonorrhoeae and the clinical doses suggest that efficacy can be translated across pathogens 
based on area under the curve (AUC)/MIC targets. From this analysis, the efficacious human 
exposure for zoliflodacin was estimated utilizing a PK/PD target that covers a mean fAUC/MIC 
of 66 (range 43-98) for S. aureus in the mouse thigh model. This target, combined with the 
MIC90 of zoliflodacin for N. gonorrhoeae and the human unbound fraction (fu) of 17%, 
translated to a predicted efficacious mean AUC in humans of 49 µg*h/mL.8,9 
In vivo: Human Phase 2 trial: Zoliflodacin was recently investigated in a Phase 2, multi-center, 
randomized trial to assess the safety and efficacy of oral zoliflodacin to treat uncomplicated 
urogenital gonorrhea in adults compared to intramuscular (IM) ceftriaxone. Subjects were 
randomly assigned to receive a single oral dose of 2 g of zoliflodacin, 3 g of zoliflodacin, or a 
single IM dose of 500 mg of ceftriaxone. In the per-protocol population, microbiological cure 
was observed in 96% and 100% in patients treated with 2 g and 3 g of zoliflodacin, respectively.8 

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Human Pharmacokinetics 
A Phase 1, single-center, multi-part trial with an oral suspension of zoliflodacin vs. placebo was 
conducted in healthy men and women. In part A, 48 subjects received single ascending doses of 
zoliflodacin ranging from 200 mg to 4 g, or placebo (6:2, zoliflodacin: placebo). In Part B, 
zoliflodacin was administered in doses of 1.5 g (N=8) and 3 g (N=10) to evaluate the effect of 
food on the PK of zoliflodacin. Zoliflodacin was absorbed relatively quickly under fasting 
conditions, with a median time to maximum plasma concentration (Tmax) ranging from 
 
 Following maximum plasma concentration (Cmax), plasma 
drug concentrations 
 The terminal elimination phase 
started at approximately 
 zoliflodacin concentrations 
 
 Exposures
 
 suggesting a 
 
 When 
 zoliflodacin was administered, 
 
 was seen if the drug was 
 The Cmax 
 
 consistent with 
 
.8 
In a Phase 2 trial of patients with gonorrhea, 2-g and 3-g doses of zoliflodacin were selected to 
maximize the probability of achieving the target plasma 
 
The AUC and Cmax values for zoliflodacin at the various doses also 
 
 values.8 
The PK of a single 4-g dose of zoliflodacin, packaged in the new formulation that will be used in 
the current trial, was evaluated in a recent Phase 1 trial in healthy subjects (DMID 16-0118) and 
was compared to PK data obtained in the previously completed trials.8,12 This analysis 
demonstrated that the PK of the 4-g dose of the new formulation of zoliflodacin is similar to the 
previous one, and higher exposure at the 4-g dose compared to the 2-g dose.   
Potential for Drug-Drug Interactions 
Zoliflodacin is a 
 in preclinical studies. In vitro and in vivo 
metabolism across species indicate that the compound is extensively metabolized via cytochrome 
P450- and non-P450-mediated pathways.
 The potential for drug-drug interactions has been evaluated using a physiologically-based 
PK (PBPK) model developed with 
.8 In summary: 
1. Zoliflodacin as a victim: Single doses of zoliflodacin ranging from 
 
 are predicted to 
 
 based upon Phase 1 PK data in healthy subjects, 
 

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 Since the 
primary PK driver for efficacy of zoliflodacin is 
. 
2. Zoliflodacin as a perpetrator: Zoliflodacin is predicted to increase 
 
 
. The extent of the interaction is not predicted to differ 
between the zoliflodacin 
. This interaction 
 
 
 administration of zoliflodacin. 
Safety 
In vitro studies: There was no
 
 There was no inhibition of 
 
.  
Some fluoroquinolones induce QT prolongation leading to arrhythmias and torsade de pointes 
(TdP), which have been correlated with binding to the hERG K+ channel. In contrast, 
zoliflodacin showed
 
 
 
 
Animal pharmacology and toxicology studies: Zoliflodacin was well-tolerated –
 
 
 There was 
 on any evaluated 
endpoints including 
 
 
were observed up to the highest dose tested. 
 
 were evident at either primary or recovery necropsy. 
 
 was established at 
 and correlated with 
 
 
At 
this highest dose tested, 
 was observed, consistent with
 
 These effects were 
 

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can be monitored in the clinical setting. There were 
 
 with zoliflodacin administration. 
 
 
, there were
 
.8 
Effects of zoliflodacin on female fertility and embryofetal development have been assessed in 
. Effects on 
  
Following administration of zoliflodacin at doses of 
 
 
 At all dose levels, there were 
 
. 
 were given zoliflodacin at 
 
 there was
 
there was 
 
 was considered to be 
. 
The effects of zoliflodacin on male fertility have been assessed in 
 were 
dosed 
. Administration at 
 
 was associated with a 
 
 which was 
 
. 
Human trials: Four trials have evaluated the safety of zoliflodacin in humans.  
In healthy subjects in the Phase 1 trials, 
 
 The most common AEs in the zoliflodacin groups were 
 
 There were 
 
.8,12 
In patients in the Phase 2 trial, zoliflodacin was generally safe and well-tolerated. Reported AEs 
were generally mild, were non-serious, and none led to trial discontinuation. The AE and 
laboratory safety profiles of zoliflodacin were generally similar to those of ceftriaxone. The AE 
profile of zoliflodacin 2 g was generally similar to that of zoliflodacin 3 g; headache was more 
common with zoliflodacin 3 g (9%) compared to zoliflodacin 2 g (0%). Dysgeusia was not 
reported in the Phase 2 trial. Vomiting was reported by one patient receiving zoliflodacin 3 g.8,13 
There were no AEs reported at a frequency of ≥5% when the AE profiles of the zoliflodacin 2-g 
and zoliflodacin 3-g groups were combined. The most common AEs reported with zoliflodacin 

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(≥3% based on the zoliflodacin 2-g and zoliflodacin 3-g groups combined and excluding 
infection-related AEs) were diarrhea, headache, and nausea.8,13 
In a recently completed Phase I open-label clinical trial (DMID 16-0118), a single 4-g dose of 
zoliflodacin formulated by the new method was shown to be safe and well-tolerated, and there 
were no differences in the safety profile from earlier studies.  
2.2 
Rationale 
2.2.1 
Study Rationale 
The current clinical trial will be conducted to assess the effect of zoliflodacin on cardiac 
repolarization. Medicinal products that prolong cardiac repolarization may have a proarrhythmic 
effect and have been associated with a specific, potentially fatal polymorphic ventricular 
tachycardia termed torsade de pointes (TdP). This tachycardia is usually observed in the setting 
of a prolonged QT interval, is often initiated after extrasystolic pauses, and is identified by the 
continuously twisting appearance of the QRS complex in the 12-lead ECG recording. Additional 
risk factors associated with a potential predisposition to the proarrhythmic effect include cardiac 
disease, congenital Long QT Syndrome (LQTS), hypokalemia, hypomagnesemia, bradycardia, 
and concurrent administration of other medicinal products known to prolong the QT interval. 
The QT interval, which represents ventricular recovery time, reflects the time between the 
depolarization of the first myocardial cells and the completion of repolarization of the last 
myocardial cells. Like ventricular refractoriness, it is influenced by the autonomic nervous 
system and changes in HR, and also by electrolyte disturbances and medications.14,15 The 
duration of the QT interval has prognostic implications in both healthy and diseased   
populations, with an abnormally prolonged QT interval becoming an increasingly accepted 
marker of risk for malignant ventricular tachyarrhythmias.16 
There are a number of compounds in various therapeutic classes that have proarrhythmic 
potential, including, among others, imidazoles, psychotropic agents, antihistamines, and 
antibiotics.17 Of particular concern is concurrent administration of other medicinal products 
known to prolong the QT interval and/or that are metabolized by and compete for cytochrome 
P4503A4 (CYP3A4) enzymes. A partial list of drugs considered to be associated with high-risk 
of TdP when associated with QTc prolongation is presented in Appendix E: A Partial List of 
Drugs Causing QTc Prolongation and Torsade de Pointes.   
Regulatory guidance has emphasized the need to obtain clear robust data on the effect of new 
chemical entities on ECG parameters with focus on cardiac repolarization as measured by the 
corrected QT interval (QTc) duration.16 Though many Phase 1, 2, and 3 studies may be 
conducted, they usually have insufficient sample size, infrequent sampling of ECG data, or the 
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This has resulted in regulatory guidance recommending a dedicated TQT study to define the 
ECG effects of new drugs.16 
Therefore, this four-arm crossover trial will include therapeutic and supratherapeutic dosing with 
zoliflodacin, placebo as a negative control, and oral moxifloxacin as a positive control. The trial 
will be conducted in accordance with relevant ICH E14 and Food and Drug Administration 
(FDA) guidance.16 
The positive control (moxifloxacin) will serve to validate the sensitivity of the assay to detect 
small increases from baseline in the QTc interval corrected using Fridericia’s formula (QTcF). 
Moxifloxacin will be used because it has been demonstrated to increase QTc by a mean of 6 
msec (time-averaged analysis: standard deviation (SD) = 26 msec; N = 787) after a 400-mg oral 
dose.16,18 However, thorough ECG studies have shown that this dose of moxifloxacin can be 
expected to show, for a time-matched analysis, a mean change of 10-15 msec, generally within 
1-6 h after dosing.19 
This trial will be performed in healthy subjects to eliminate variables known to change ECG 
parameters (e.g., concomitant drugs or diseases) among subjects. The sample size is driven by 
the need to do a time-matched statistical analysis on the primary endpoint as per ICH E14 
guidance. A crossover study design was chosen so that subjects will act as their own controls. A 
crossover trial can be conducted since the half-life of zoliflodacin is approximately 6 h. A wash-
out phase of 8 ±2 days after Day 1 of each dosing period is considered sufficient to prevent a 
carryover effect of the study drug between treatments. 
2.2.2 
Rationale for Dose Selection 
The zoliflodacin 2-g dose chosen for this trial is intended to achieve the drug exposure achieved 
in gonorrhea patients for the anticipated therapeutic single dose of 2-3 g. Several elements were 
considered in the selection of the zoliflodacin supratherapeutic dose including the anticipated 
clinical usage, the Phase 1 data with the original formulation, including utilization of population 
PK modeling and PBPK modeling, the potential for drug-drug interactions to influence 
zoliflodacin Cmax, and lack of availability of an IV formulation: 
1. Anticipated clinical usage: Zoliflodacin is being studied as a single-dose oral therapy for 
the treatment of uncomplicated gonorrhea. The product is not intended for multiple-dose 
administration, which has not been studied. The Cmax for zoliflodacin 4 g is anticipated to 
be supratherapeutic to peak concentrations observed for a therapeutic dose of 2-3 g when 
the drug is taken in the fasted state. 
2. Phase 1 data including utilization of population PK modeling and PBPK modeling: 
In a Phase 1 study, in the dose range
 
 
 The population PK model that has been developed adequately described the 
clinical data observed in healthy subjects 

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.8 
 
. Based on the 
population PK model, therefore, 
 
 
3. PBPK modeling: In addition to the population PK model, a PBPK model incorporating 
physicochemical properties, absorption, distribution, and elimination of zoliflodacin was 
developed to investigate the systemic exposure of the drug after oral administration to 
healthy adults and determine the potential for drug-drug interactions 
 
8. The PBPK model was utilized to assess 
 
. The model 
 
 
. Based on these simulations, 
 
  
4. Potential for drug-drug interactions to influence zoliflodacin Cmax levels: Using the 
PBPK model 
8, the potential for zoliflodacin to be 
either a victim and/or a perpetrator in drug-drug interactions was evaluated. Although 
metabolic clearance of zoliflodacin is mediated predominantly by 
 
 
 
. Based on these simulations, 
 
. 
5. The only formulation under development is the oral formulation: There is no IV 
formulation available for use in the TQT study. 
In the recently completed Phase 1 trial assessing PK parameters in healthy subjects 
 
. Thus, the 4-g dose will be used as the supratherapeutic dose required by the ICH 
E14 Guidance for TQT studies.16 Both the 2-g and 4-g doses of zoliflodacin were shown to be 
safe in previous studies. The 400-mg dose of moxifloxacin to be used is the highest dose 
recommended for use in the clinic and is commonly used as a positive control in TQT studies to 
ensure assay sensitivity for detecting potentially clinically significant changes in the QT/QTc 
interval.16,20,21  
2.3 
Identified and Potential Risks and Benefits 
2.3.1 
Zoliflodacin Risks 
 

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Identified Risks 
There are no identified risks associated with zoliflodacin to date. Single zoliflodacin doses 
ranging from 200 mg to 4 g were generally safe and well-tolerated in the Phase 1 and Phase 2 
clinical trials conducted to date in healthy subjects and patients with uncomplicated 
gonorrhea.8,12,13 The most common AEs reported with zoliflodacin in the Phase 1 trials were 
dysgeusia and headache, and in the Phase 2 trial (≥3% based on the zoliflodacin 2-g and 
zoliflodacin 3-g groups combined and excluding infection-related AEs) were diarrhea, headache, 
and nausea. These events will continue to be monitored and evaluated. 
Potential Risks 
As for any drug, exposure to zoliflodacin could result in an allergic or hypersensitivity reaction 
that could be mild or life-threatening. Should a reaction occur, medically-appropriate diagnostic 
and therapeutic measures will be taken immediately by trained site clinicians and staff. 
In a Phase 1 trial, analysis of 
 
 
Continuous monitoring of 
 will be implemented in this trial. 
In a Phase 1 and Phase 2 trial, no trends in
 
 
. Standard clinical monitoring 
 
. 
 
 
. The incidence of these 
findings typically 
 
 
 
 
 
 in the first-in-human trial. 
 
.  
Effects of chronic dosing of zoliflodacin on 
 
 With respect to 
 
 
 
 
 
 

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. The effects of zoliflodacin 
 
 
. Based on these preclinical data and the 
gonorrheal clinical indication, in current and future clinical trials: 
• Female subjects will be pregnancy tested before administration of zoliflodacin and at the 
end of the trial and will be required to use specified birth control methods during the trial 
and for 30 days after Final Visit.  
• Male subjects will be required to use specified birth control methods during the trial and 
for 90 days after Final Visit and will be advised not to donate sperm during this period.  
2.3.2 
Moxifloxacin Risks 
Moxifloxacin is a broad-spectrum 8-methoxy fluoroquinolone with activity against both Gram-
positive and Gram-negative bacteria, including anaerobes. It is a licensed antibiotic for serious 
and life-threatening bacterial infections and can be used when other drugs fail. The maximum 
recommended dosage is 400 mg daily.18, 22  
Overdose: Single oral overdoses up to 2.8 g were not associated with any SAEs. In case of acute 
overdose, the stomach should be emptied, and adequate hydration maintained. ECG monitoring 
is recommended due to the possibility of QT interval prolongation, and the patient should be 
carefully observed and given supportive treatment. The administration of activated charcoal as 
soon as possible after oral overdose may prevent excessive systemic moxifloxacin exposure.  
Adverse reactions: AEs judged by investigators to be at least possibly drug-related, occurring in 
≥2% of moxifloxacin-treated patients, were nausea (6%), diarrhea (5%), and dizziness (2%).  
Electrocardiogram: Prolongation of the QT interval in the ECG has been observed in some 
patients receiving moxifloxacin. Following oral dosing with 400 mg of moxifloxacin the mean (± 
SD) change in QTc from the pre-dose value at the time of maximum drug concentration was 6 
msec (± 26) (n = 787). Following a course of daily IV dosing (400 mg; 1-h infusion each day), 
the mean change in QTc from the Day 1 pre-dose value was 9 msec (± 24) on Day 1 (n = 69) and 
3 msec (± 29) on Day 3 (n = 290). No cardiovascular morbidity or mortality attributable to QTc 
prolongation occurred with moxifloxacin treatment in over 9,200 patients in controlled clinical 
trials, including 223 patients who were hypokalemic at the start of treatment, and there was no 
increase in mortality in over 18,000 moxifloxacin tablet-treated patients in a post-marketing 
observational study in which ECGs were not performed. 
In post-marketing studies, rare AEs were reported in patients treated with moxifloxacin. These 
include the following: 
Tendinopathy and tendon rupture: Fluoroquinolones, including moxifloxacin, are associated 
with an increased risk of tendinitis and tendon rupture in all ages. This AE most frequently 

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involves the Achilles tendon, and rupture of the Achilles tendon may require surgical repair. 
Tendon rupture can occur during or after completion of therapy; cases occurring up to several 
months after completion of therapy have been reported. 
Photosensitivity: Photosensitivity, which can appear as skin eruption or severe sunburn, can 
occur in some patients taking quinolone antibiotics after exposure to sunlight or artificial 
ultraviolet light (e.g., tanning beds).   
Central nervous system: Quinolones may cause central nervous system events including 
convulsions, dizziness, confusion, tremors, hallucinations, depression, and, rarely, suicidal 
thoughts or acts. Nervousness, agitation, insomnia, anxiety, nightmares or paranoia have also 
been reported. These reactions may occur following the first dose. 
Subjects will be closely monitored for all of these risks. 
2.3.3 
Venipuncture and IV Catheter Placement Risks 
Venipuncture: Venipuncture causes transient discomfort and may cause fainting. Bruising at the 
site of venipuncture may occur but can be prevented or lessened by applying pressure for several 
min. Infection at the site is possible but highly unlikely as aseptic technique will be used. 
IV catheter placement: An indwelling catheter may be placed in an arm vein (preferably 
antecubital) for frequent blood drawing for PK measurements. The catheter may cause phlebitis 
with signs of redness and warmth at or near the IV insertion site, and thrombophlebitis with a 
hard area palpable near the IV insertion site. These risks are minimal as the IV catheters, when 
used, are only used briefly after dosing. Careful inspection of the catheter site, including 
visualization of blood return, and withdrawal of the catheter if needed will minimize this risk. 
There is a risk of infection; however, this is a small risk as aseptic technique will be used. 
2.3.4 
Additional Risks 
ECG: Possible side effects from ECG patches include a rash or minor irritation of the skin. 
Blood draws: The amount of blood drawn is about 17.5 mL at Screening Visit, 9 mL on Day -1, 
69 mL during each inpatient period (Days 1-2), 9 mL at Final Visit, and 338.5 mL during the 
entire trial (Appendix D). Additionally, small amounts of blood loss may occur if an IV catheter 
is used or additional blood samples are collected (for repeat laboratory testing, AE evaluation, 
etc.). Overall, the amount of blood that may be drawn during the trial is within the amount that 
considered safe to be drawn during short or extended periods, respectively, and not excessive for 
the safety and PK assessment requirements of Phase 1 trials. However, there is a small risk that 
some subjects may develop mild symptoms of hypovolemia or anemia during the trial. These are 
reversible with specific treatments (e.g., fluid replacement, good nutrition, vitamins, or iron 
supplementation). 

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2.3.5 
Known Potential Benefits 
The trial has no direct benefit for subjects participating in the trial. Knowledge gained in the trial 
could be of benefit to public health and to individuals with gonorrhea or at risk of acquiring it. 
In accordance with the Health Insurance Portability and Accountability Act (HIPAA) regulation, 
subjects will be promptly notified of any clinical test results that would have an impact on their 
health. Screening evaluations may detect previously unknown abnormalities that could be 
clinically significant. This could benefit a subject, as it may lead to earlier diagnostic evaluation 
and treatment of an underlying disorder. 
2.3.6 
Risk/Benefit Ratio 
This ratio cannot be calculated by available data. The risk for an SAE, including a significant 
ventricular arrhythmia, occurring in a subject treated with single doses of 2 g and 4 g of 
zoliflodacin or 400 mg of moxifloxacin is thought to be low. Subjects will be treated in an 
inpatient unit and closely monitored during the trial, so that emergency care can be provided 
immediately if an acute arrhythmia or other event occurs. In addition, should a significant QTcF 
prolongation occur, with or without TdP, the subject will receive appropriate treatment if needed, 
and counseled to obtain further consultation to rule out a genetic predisposition towards 
development of LQTS after using licensed drugs. Data collected in the clinical development of 
zoliflodacin to date, especially preliminary data in Phase 2 studies in patients with uncomplicated 
gonorrhea, support further development of zoliflodacin. The benefit of a potential new addition 
to the drugs available for the treatment of gonorrhea outweighs the known risks of the 
medications to be used in this trial.

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3 
OBJECTIVES AND OUTCOME MEASURES 
3.1 
Study Objectives 
3.1.1 
Primary 
• To evaluate the effect of zoliflodacin on the corrected QT interval of the ECG using 
Fridericia’s formula (QTcF)  
3.1.2 
Secondary 
• To evaluate the effects of zoliflodacin on other ECG parameters (PR, QRS, and RR 
intervals, and HR)  
• To evaluate the sensitivity of QTcF measurement using moxifloxacin 
• To evaluate the effect of zoliflodacin on T-wave morphology 
• To evaluate the PK of 2 g and 4 g oral zoliflodacin under fasting state 
• To evaluate the relationship between zoliflodacin PK and time-matched QTcF PD 
• To evaluate the safety and tolerability of 2 g and 4 g oral zoliflodacin 
3.2 
Study Outcome Measures 
3.2.1 
Primary 
• The upper bound of the one-sided 95% confidence interval (CI) for the largest time-
matched, placebo-corrected, baseline-adjusted mean QTcF interval (∆∆QTcF) collected 
in the 24-h period after 2 g and 4 g dose of zoliflodacin is <10 msec 
3.2.2 
Secondary 
• Time-matched, placebo-corrected, baseline-adjusted non-QT intervals (PR, QRS and RR 
intervals) and HR collected in the 24-h period after 2 g and 4 g dose of zoliflodacin 
• The lower bound of the one-sided 95% CI of the ∆∆QTcF is >5 msec at one or more 
individual timepoints between 1 h and 4 h after a single dose of moxifloxacin 400 mg  
• Incidence of abnormal T-wave morphology using defined categories in the 24-h period 
after 2 g and 4 g dose of zoliflodacin 
• Single-dose plasma concentrations (and PK exposure parameters, such as Cmax and AUC) 
of zoliflodacin collected up to 24 h after 2 g and 4 g dose of zoliflodacin 
• Relationship between plasma concentrations of zoliflodacin (and PK and exposure 
parameters, such as Cmax and AUC) and ∆∆QTcF up to 24 h after 2 g and 4 g dose of 
zoliflodacin 
• Occurrence of treatment-emergent SAEs, other unsolicited treatment-emergent AEs, and 
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administration of zoliflodacin and moxifloxacin from time of dosing to Final Visit, or ET 
Visit 
 

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4 
STUDY DESIGN 
The TQT study will be performed in a single center, the Vince & Associates Clinical Research, 
Inc., clinical trials unit (CTU), according to a randomized, double-blinded (except for the use of 
moxifloxacin), placebo-controlled, four-period, four-treatment, crossover design balanced with 
respect to first-order carryover effect in 72 healthy male or female subjects, aged 18 to 45 years 
inclusive, to evaluate the effect of zoliflodacin on QTcF and other ECG parameters, the 
correlation of the drug concentrations (and PK profile) with ∆∆QTcF, and the PK and safety 
profiles of the new zoliflodacin formulation.  
Subjects who consent to participate will be enrolled in the trial if they meet all inclusion and no 
exclusion eligibility criteria. Laboratory assessments at Screening Visit will include routine 
clinical laboratory tests (HEM, CHEM, and UA), testing for viral serology (HIV antibody, 
HBsAg, and HCV antibody), drugs of abuse, and alcohol. Serum pregnancy testing will be done 
in all women and serum FSH levels will be measured only in post-menopausal women.  
Each subject will receive one dose of each of four treatments, labeled A through D: 
• Treatment A: zoliflodacin 2 g PO (investigational drug, therapeutic dose) 
• Treatment B: zoliflodacin 4 g PO (investigational drug, supratherapeutic dose) 
• Treatment C: placebo PO (for zoliflodacin 4 g) 
• Treatment D: moxifloxacin 400 mg PO (positive control comparator) 
In the dosing phase, each subject will be randomly allocated to one of 12 dosing sequences 
before dosing on Day 1, Dosing Period 1, and receive a single dose of the following four study 
drugs on different days according to their randomly-assigned dosing sequence, as follows: 
Table 1: Dosing Sequences 
ABCD 
ACDB 
ADBC 
BADC 
BDCA 
BCAD 
CDAB 
CABD 
CBDA 
DCBA 
DBAC 
DACB 
Six of the planned 72 subjects will be included in each treatment sequence. The design is 
balanced for period and first-order carryover effects. 
Subjects will be dosed in the morning of Day 1 in each dosing period, in a staggered fashion at 
least several minutes apart, and will receive study drug at approximately the same time in each 

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subsequent dosing period. Each treatment will be administered after at least an 8-h fast, which 
will continue for at least 4 h after dosing. Consumption of water will be permitted during the 
fasting period. Zoliflodacin or placebo will be reconstituted in 60 mL of tap water. After the cup 
containing the 60 mL of zoliflodacin or placebo is taken, approximately 60 mL of tap water will 
be added to the cup and consumed by the subject to chase the initial dose. Moxifloxacin will be 
administered orally with 120 mL of water.  
Safety assessments post-dosing will include VS, PE, 12-lead standard ECG, and clinical 
laboratory tests including HEM, CHEM, and UA. Treatment-emergent AEs and SAEs will be 
assessed from time of dosing to end of trial. 
Continuous 12-lead Holter ECGs will be performed on Day 1 starting approximately 1 h before 
dosing and lasting 25 h. At timepoints for ECG extraction (see Section 8.3.1), subjects will be 
resting in the supine position for at least 10 min before and 5 min after the nominal time, without 
any ongoing stimulation (e.g., television). Three replicate ECGs will be extracted at each 
timepoint to measure ECG intervals and assess T-wave morphologies. 
Plasma for PK analysis will be collected at scheduled timepoints that match timepoints for ECG 
extraction before and after dosing on Day 1 and through Day 2, or ET if it occurs within 24 h 
after dosing. 
Subjects will be monitored as inpatients in the CTU up to 24 h after each dose (Day 2). They will 
then be discharged from the inpatient CTU and followed as outpatients for at least 5 days (Days 
3-7) after dosing in Dosing Periods 1-3. The total time to wash-out of the previously-received 
study drug is at least 8 days. The Final Visit will be 8 ±2 days after the last dose. 
Safety Monitoring and SMC Role 
A SMC will be appointed to oversee the safe conduct of the trial. A scheduled SMC meeting will 
be held after all subjects complete the trial to review safety data and advise on safety monitoring 
in subsequent clinical trials with zoliflodacin. If criteria for halting the trial (as listed in Section 
9.5.1) are met in any dosing period, dosing will be suspended, and an ad hoc SMC meeting will 
be held to review all available safety data and to make recommendations about the dosing of all 
further subjects in the trial. An independent safety monitor (ISM), who is local to the CTU, will 
review SAEs and other severe safety signals and provide an independent analysis to the Site 
principal investigator (PI), SMC, and DMID. 
4.1 
Sub-studies 
No sub-studies are planned. 

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5 
STUDY ENROLLMENT AND WITHDRAWAL 
Only subjects who meet all inclusion criteria and no exclusion criteria will be eligible for 
enrollment into the trial. No exemptions are granted on Inclusion/Exclusion Criteria in DMID-
sponsored trials. 
72 healthy male and female subjects, aged 18 to 45 years inclusive, will be enrolled in the trial.  
5.1 
Subject Inclusion Criteria 
All must be answered YES for the subject to be eligible for study participation: 
1) Informed consent form (ICF) understood and signed before initiating any study 
procedures 
2) Healthy male or female, as assessed by authorized site clinician (listed on FDA Form 1572)  
3) Willingness to comply with and be available for all protocol procedures, including 
inpatient confinement for 3 days in each dosing period and follow-up for the duration of 
the trial 
4) Aged 18 to 45 years inclusive on the day of first dosing 
5) Body Mass Index (BMI) ≥18.5 and ≤30 kg/m2 and weight ≥50 kg (110 lbs.) and 
≤100 kg (220 lbs.) 
6) In all female subjects, whether of childbearing potential or post-menopausal by MH, a 
negative serum pregnancy test at Screening Visit and on Day -1 of each dosing period 
• Note: A woman is considered of childbearing potential unless post-menopausal (≥1 
year without menses without other known or suspected cause and with a FSH level in 
the menopausal range), or surgically sterilized (hysterectomy, salpingectomy, 
oophorectomy, or tubal ligation/occlusion). 
7) If female, not pregnant, not breast feeding, and not planning to become pregnant during 
the trial and for 30 days after Final Visit 
8) Females of childbearing potential and males agree to use acceptable contraception for the 
duration of the trial and for 30 days (females) or 90 days (males) after Final Visit 
• Note: A highly effective method of birth control is defined as one with a low failure 
rate (i.e., <1% per year) according to CDC criteria.23 These include progestin 
implants, intrauterine devices (IUDs), surgical (hysterectomy, salpingectomy, 
oophorectomy, or tubal ligation/occlusion; vasectomy), or abstinence. Use of 
methods with higher failure rate (such as progestin injectables, combined oral 
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used alone, but they could be considered if used in combination with another method 
(e.g., a female using combined oral contraceptives if her male partner is sterile, or if 
she and her non-sterile male partner use a double-barrier method), after consultation 
with the DMID Medical Officer. 
9) Male subjects agree to refrain from sperm donation for the duration of the trial and for 90 
days after Final Visit 
10) Laboratory tests, as outlined in Section 8.2, are in the normal reference range with 
acceptable exceptions as noted in Section 8.2.1 and Appendix B  
11) VS, as outlined in Section 8.1.6, are within the acceptable range per Appendix B 
12) Has adequate venous access for blood collection  
13) Urine drug screen is negative for tested substances (see Section 8.2.5) 
14) Urine alcohol test is negative 
15) Willing to abstain from alcohol consumption for 2 days before Day -1 of Period 1 and for 
the duration of the trial 
5.2 
Subject Exclusion Criteria 
All must be answered NO for the subject to be eligible for study participation: 
1) History of acute or chronic cardiovascular disease or surgery 
• Note: Conditions include: congestive heart failure; coronary artery disease 
(myocardial infarction, unstable angina); cerebrovascular disease ( cerebrovascular 
accident or stroke or transient ischemic attack (TIA); chronic hypertension; or 
coronary revascularization surgery (coronary artery bypass grafting or percutaneous 
transluminal coronary angioplasty) 
2) History of cardiac arrhythmia or syncope related to cardiac arrhythmia or unexplained, or 
use of a cardiac pacemaker  
• Note: Conditions include: atrial fibrillation, atrial flutter, or non-sustained or 
sustained ventricular tachycardia); use of a cardiac pacemaker; personal or family 
history of LQTS; or family history of sudden death    
3) History of any other chronic medical or surgical condition that would interfere with the 
accurate assessment of the trial’s objectives or increase the subject’s risk profile 
• Note: Chronic medical conditions include: diabetes mellitus; asthma requiring use of 
medication in the year before screening; autoimmune disorder such as lupus 
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low-grade (squamous and basal cell) skin cancer thought to be cured; chronic renal, 
hepatic, pulmonary, or endocrine disease, myopathy, or neuropathy; gastrointestinal 
surgery including weight loss surgery or biliary surgery 
4) Major surgical interventions are not permitted within 4 weeks of first dosing and during 
the trial. Minor surgical interventions are not allowed within 2 weeks of first dosing and 
during the trial. 
5) History of hypersensitivity or severe allergic reaction of any type to medications, bee 
stings, food, or environmental factors 
• Note: Severe allergic reaction is defined as any of the following: anaphylaxis, 
urticaria, or angioedema 
6) Active allergic symptoms to seasonal and animal allergens that are moderate to severe, 
affect daily activity, and require continuous treatment   
7) A marked baseline prolongation of ECG intervals, or HR <45 bpm or >100 bpm on ECG 
measurements 
• Note: The following are considered prolonged ECG intervals: QTc/QTcF >449 msec 
in males and females; PR >209 msec; and QRS >110 msec 
8) Clinically significant abnormal ECG results 
Note: Clinically significant abnormal ECG results include: complete left or right 
bundle branch block; other ventricular conduction block; 2nd degree or 3rd degree 
atrioventricular (AV) block; sustained atrial or ventricular arrhythmia; two premature 
ventricular contractions in a row; pattern of ST elevation felt consistent with cardiac 
ischemia; evidence of a previous myocardial infarction (MI), left ventricular 
hypertrophy (LVH), or more than minor non-specific ST-T wave changes; any 
characteristics that would make QT assessment unreliable, including flat T waves; or 
any condition deemed clinically significant by a study investigator 
9) Abnormal renal function  
Note: Normal renal function is defined as normal creatinine [per criteria in Appendix B] 
and normal estimated glomerular filtration rate (eGFR) [i.e., >80.0 mL/min] values 
according to Cockroft-Gault 
10) Positive serology results for HIV, HBsAg, or HCV 
11) Febrile illness with temperature ≥38.0°C for <7 days before dosing in each treatment 
period 
12) Donated whole blood or blood products within 60 days before first dosing, or plans to 
donate or receive before Final Visit (Day 8 ±2 after last dose in dosing period 4) 

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•  Note: Blood products include RBCs, white blood cells (WBCs), platelets, and plasma 
13) Known allergic reactions to fluoroquinolones or to components present in the formulation 
or processing of zoliflodacin and moxifloxacin 
14) Treatment with another investigational product within 30 days of first dosing or 5 half-
lives or twice the duration of the biological effect of the study drug (whichever is longer)  
• Note: Investigational products include a drug, vaccine, biologic, device, or blood 
product 
15) Active drug or alcohol binge consumption, abuse, or dependence within 12 months 
before Screening Visit that, in the opinion of the investigator, would interfere with 
adherence to study requirements 
16) Use of any prescription medication within 30 days before first dosing or planned use 
during the trial except as noted below and approved by the designated study clinician 
• Note 1: Prohibited medications include moderate or strong CYP3A4 inducers (per 
6.6) and other drugs with known risk for QT prolongation and TdP (Appendix E and 
17); antibiotics; injectable or oral antidiabetic drugs; anti-lipid drugs; 
immunosuppressive agents; immune modulators; oral corticosteroids; anti-neoplastic 
agents; any vaccine (licensed or investigational) except licensed influenza vaccine 
during the flu season, which is allowed up to 7 days before first dosing or 7 days after 
last dosing 
• Note 2: Allowed medications include: oral contraceptives; H1 antihistamines; all 
medications approved for control of intraocular pressure including topical 
ophthalmic non-selective β-blockers, such as betaxolol, carteolol, levobunolol, 
metipranolol, and timolol; topical/ intranasal corticosteroids; nonsteroidal anti-
inflammatory drugs (NSAIDS); licensed influenza vaccine during the flu season, 7 
days before first dosing or 7 days after last dosing 
17) Use of non-prescription medications, vitamins, herbs, or nutritional supplements within 
15 days before first dosing or planned use during the trial unless approved by the study 
clinician 
• Note 1: Intake of nutritional supplements, juice, and herbal preparations or other 
foods or beverages that may affect the various drug-metabolizing enzymes and 
transporters (e.g., grapefruit, grapefruit juice, grapefruit-containing beverages, apple 
or orange juice, vegetables from the mustard green family [e.g., kale, broccoli, 
watercress, collard greens, kohlrabi, Brussels sprouts, mustard], and charbroiled 
meats) within 7 days before dosing 
• Note 2: Exceptions: St. John’s wart is not allowed within 30 days of dosing; vitamins 

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and over-the-counter (OTC) medications taken for a brief period (<48 h) for the 
treatment of common symptoms (such as headache, indigestion, muscle pain) may be 
allowed as approved by the designated study clinician 
18) Intake of caffeinated beverages or food within 72 h before first dosing or a history of high 
caffeine consumption (e.g., in the last 4 months drinking >5 cups of coffee/day) 
19) Smoking or use of tobacco or nicotine-containing products within 30 days before first 
dosing 
20) Engagement in strenuous exercise within 15 days before first dosing (e.g., marathon 
running, long-distance cycling, weight lifting) and during the trial 
21) Any specific behavioral or clinical condition that, in the judgment of the investigator, 
precludes participation because it could affect compliance with study procedures or 
subject safety 
22) Plans to enroll or is already enrolled in another clinical trial that could interfere with 
safety assessment of the study drug at any time during the trial 
• Note: Includes trials that have a study intervention such as a drug, biologic, or device 
23) Is a CTU employee or staff member who is paid entirely or partially by the Office of 
Clinical Research Resources (OCRR)/NIAID contract for the DMID-funded trial 
• Note: CTU employees or staff include the PIs, sub-investigators, or staff who are 
supervised by the PI or sub-investigators 
5.3 
Treatment Assignment Procedures 
5.3.1 
Enrollment and Randomization Procedures 
Seventy-two healthy subjects who consent to participate in the trial and meet the eligibility 
criteria will be enrolled following admittance to the CTU and confirmation of eligibility. 
Subjects will be registered using a web-based application developed by The Emmes Corporation, 
the Data Coordinating Center (DCC) for the trial. 
Randomized sequence assignments will be generated by a statistician at the DCC for this study.  
Randomization will occur following admittance to the CTU and confirmation of eligibility is 
confirmed. 
Subjects will be randomized to one of 12 treatment sequences (Section 4) with equal allocation 
(1:1:1:1:1:1:1:1:1:1:1:1). The randomization list will be generated centrally through the 
AdvantageEDCSM (Electronic Data Capture [EDC] System, Emmes) by the unblinded study 
biostatistician, and a list will be transferred to the unblinded study pharmacist before starting the 
study for the purpose of an emergency back-up. 

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Per ICH guideline E6: GCP, screening records will be kept at the CTU to document the reason 
why an individual was screened but did not meet trial entry criteria, by recording it in 
AdvantageEDCSM. 
Enrollment of subjects will be done online using the enrollment module of AdvantageEDCSM.  
The randomization code will be prepared by statisticians at the DCC and included in the 
enrollment module for this trial. AdvantageEDCSM will assign each subject to a treatment 
sequence after the demographic and eligibility data have been entered into the system. A 
designated individual at the participating site will be provided with a code list for emergency 
unblinding purposes, which will be kept in a secure place. Instructions for using the enrollment 
module are included in the AdvantageEDCSM User’s Guide. 
5.3.2 
Masking Procedures 
This is a double-blinded clinical trial with respect to administration of zoliflodacin or placebo. 
The study staff administering the study drug and assessing subjects will not be aware of the 
administered zoliflodacin or placebo. Administration of moxifloxacin will be open-label.  
The SMC will review aggregate data in open session and review grouped data only in closed 
session. 
5.3.3 
Reasons for Withdrawal and Discontinuation of Study Drug Administration 
A subject may withdraw from the trial at any time for any reason, without any consequences. 
A subject will be discontinued from the trial if any of the following occur before dosing: 
• Request by the subject to terminate participation; 
• Failure to receive the study drug due to difficulty ingesting it. 
A subject may be removed from the trial after dosing for the following reasons; however, 
whenever possible, the subject will be followed for safety per protocol: 
• Failure to adhere to protocol requirements; 
• Loss to follow-up; 
• Request of primary care provider; 
• Request of the Institutional Review Board (IRB)/Ethics Committee (EC), FDA, or 
DMID; 
• The subject’s well-being, based on the opinion of the investigator; 
• The occurrence of QTcF interval prolongation >500 msec or ≥60 msec above baseline 
or occurrence of a symptomatic or asymptomatic ventricular tachyarrhythmia with or 
without QTcF prolongation;  
• The occurrence of an SAE or AE warranting withdrawal. 
 

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A subject may be removed from data analysis for a dosing period during which the following 
events occurred, but not withdrawn from the trial (see also Section 5.3.4): 
• Failure to ingest the entire 60 mL volume of zoliflodacin suspension or placebo 
suspension; 
• Vomiting up to 5 h after dosing with zoliflodacin or placebo or moxifloxacin. 
5.3.4 
Handling of Withdrawals and Discontinuation of Administration 
• Subjects who are withdrawn before dosing may be replaced with a subject assigned 
the same sequence.  
• A subject who cannot ingest the entire 60 mL volume of zoliflodacin or placebo 
suspension or a subject who took the entire dose but vomits up to 5 h after dosing 
with zoliflodacin or placebo or moxifloxacin will be withdrawn from data analysis for 
the period when the event occurred, but will not be withdrawn from the trial and may 
complete the remaining treatment periods in the assigned dosing sequence.  
• Subjects who ingest the entire volume of the initial 60-mL drug suspension but who 
do not ingest the content of the second 60-mL rinse will not be withdrawn from the 
trial.  
• Subjects who withdraw after receiving at least one treatment in a treatment sequence 
will not be replaced, unless more than 16 subjects withdraw from the trial.  
• Subjects who received any amount of the study drug but withdraw from the trial 
within 24 h after dosing will be encouraged to continue follow-up (with subjects’ 
consent) for safety assessments and PK sample collection.  
• Subjects withdrawing will be asked to complete an ET Visit if they do not wish to be 
followed per protocol. 
5.3.5 
Lost to Follow-up 
If subjects fail to appear for a follow-up assessment, extensive effort (i.e., three documented 
contact attempts via phone calls, e-mail, etc., made on separate occasions and followed by a 
certified letter) will be made to locate or recall them, or at least to determine their health status. 
These efforts will be documented in the subjects’ records. 
5.3.6 
Termination of Study 
Although DMID has every intention of completing the trial, it reserves the right to terminate the 
trial at any time for clinical or administrative reasons. In addition, the trial may be terminated or 
suspended at the request of the FDA, SMC, or IRB/EC. 

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6 
STUDY INTERVENTION/INVESTIGATIONAL 
PRODUCT 
6.1 
Description of Study Products  
Zoliflodacin will be the investigational drug in this clinical trial and its effects will be compared 
to placebo matching the 4-g dose. Moxifloxacin will be the positive control (comparator) drug. 
Zoliflodacin 
The drug product is presented as a powder for oral suspension in a single sachet, each containing 
2 g of zoliflodacin, a spiropyrimidinetrione antibacterial agent. The sachet contains both 
zoliflodacin and excipients. The excipient ratio for zoliflodacin is 
 
; the excipients are detailed in the IB and Manual of 
Procedures (MOP) or Pharmacy Manual. Further details regarding the method of reconstitution 
and administration will be provided in the MOP or Pharmacy Manual. 
Zoliflodacin placebo  
This is composed of 4 g of the same excipients found in the drug 
 
 
. The excipients are detailed in the IB and the MOP or Pharmacy Manual.  
Moxifloxacin Hydrochloride  
Moxifloxacin is a slightly yellow-to-yellow crystalline substance, formulated for human use as 
400-mg film-coated tablets. It is a synthetic broad-spectrum, fluoroquinolone antibacterial agent 
that is licensed for oral administration as an antibiotic. 
Moxifloxacin is available as the monohydrochloride salt of 1-cyclopropyl-7-[(S,S)-2,8-
diazabicyclo [4.3.0]non-8-yl]-6-fluoro-8-methoxy-1,4-dihydro-4-oxo-3-quinoline carboxylic 
acid.18 The excipients include cellulose microcrystalline, croscarmellose sodium, magnesium 
stearate, ferric oxide red, ferric oxide yellow, hypromellose 2910, polyethylene glycol 400, 
polysorbate 80, sodium stearyl fumarate, talc, silicon dioxide, and titanium dioxide. 
6.1.1 
Acquisition 
Zoliflodacin sachets and placebo are currently stored at a Good Manufacturing Practice (GMP) 
facility. Upon request by DMID, they will be shipped to the following address:    
DMID-Clinical Materials Services (CMS) 
Fisher BioServices 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 

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Tel: 240-477-1350 
Fax: 240-477-1360 
E-mail: DMID.CMS@ThermoFisher.com  
The study drugs will be shipped from DMID-CMS to the CTU upon request and approval by 
DMID. Details will be provided in the protocol-specific MOP. 
Moxifloxacin hydrochloride tablets, 400 mg, NDC 55111-0112-30 , will be purchased from the 
manufacturer, Dr. Reddy’s Laboratories Limited, Bachupally – 500 090, India, and will be stored 
at DMID-CMS before shipment to the CTU.  
6.1.2 
Formulation, Packaging, and Labeling 
Zoliflodacin 
Zoliflodacin is presented as granules for oral suspension, 50% wt/wt, packaged in a single 
sachet. Each sachet contains 2 g of spray-dried zoliflodacin,
 
 
. 
Each sachet will be labeled in compliance with applicable regulatory requirements, including the 
FDA-required cautionary statement, “Caution – New drug – Limited by Federal (or United 
States) Law to Investigational Use.”  
Zoliflodacin Placebo 
The placebo is provided in powder form as a bulk mixture of the drug product formulation but 
excluding zoliflodacin. 
Moxifloxacin 
Moxifloxacin is available in a bottle containing 30 unit doses of film-coated tablets of 
moxifloxacin hydrochloride USP equivalent to 400 mg of moxifloxacin.  
6.1.3 
Product Storage and Stability 
Zoliflodacin  
The zoliflodacin granules for oral suspension, 50% (w/w), will be stored at 2-8°C in the primary 
packaging.  
Zoliflodacin Placebo 
The placebo will be stored at 20-25°C. 
Moxifloxacin 
Moxifloxacin tablets will be stored at 20-25°C (68-77°F). A high moisture (humidity) storage 
environment will be avoided. 

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The clinical supplies storage area at the CTU will be monitored by its staff for temperature 
consistency with the acceptable storage temperature range specified in this protocol or in the 
Manual of Operations. Documentation of temperature monitoring will be maintained. 
6.2 
Dosage, Preparation, and Administration of Study Intervention/ 
Investigational Products 
Treatment A: One sachet containing 2 g of zoliflodacin will be reconstituted in 60 mL of tap water by 
the unblinded pharmacist and dosed orally after an overnight fast. The dose will be administered as a 
mix-and-drink solution in a standardized cup as described in the MOP. After the cup containing 60 mL 
of zoliflodacin is taken, approximately 60 mL of tap water will be added to the cup and consumed by 
the subject to chase the initial dose. 
Treatment B: Two sachets containing 2 g of zoliflodacin each will be reconstituted in 60 mL of 
tap water by the unblinded pharmacist and dosed orally after an overnight fast. The dose will be 
administered as a mix-and-drink solution in a standardized cup as described in the MOP. After 
the cup containing 60 mL of zoliflodacin suspension is taken, approximately 60 mL of tap water 
will be added to the cup and consumed by the subject to chase the initial dose.  The second 60 
mL volume is called ‘the rinse”.  
Treatment C: A single dose of 4 g of placebo in powder form, whose quantity will match that in 
the 4-g dose of zoliflodacin, will be reconstituted in 60 mL of tap water by the unblinded 
pharmacist and dosed orally after an overnight fast. The dose will be administered as a mix-and-
drink solution in a standardized cup as described in the MOP. After the cup containing 60 mL of 
placebo is taken, approximately 60 mL of tap water will be added to the cup and consumed by 
the subject to chase the initial dose. The second 60 mL volume is called ‘the rinse”.  
Treatment D: A single, commercially-available, film-coated, 400-mg tablet of moxifloxacin 
hydrochloride will be administered orally with 120 mL of tap water.  
Dosing, which includes ingestion of the entire 120 mL of tap water for any of the treatments (60 
mL of zoliflodacin or placebo suspension and 60 mL of the rinse, or 120 mL for taking the 
moxifloxacin tablet), will be completed within 5 min of initial suspension for treatment A, B, and 
C and ingestion for treatment D by a subject. After dosing, subjects will continue to fast for an 
additional 4 h while water is allowed ad lib. 
The CTU unblinded pharmacists will complete appropriate documentation such that the 
reconstituted materials dispensed to the CTU are clearly identified in a document suitable for 
auditing and monitoring. Clinical staff will also maintain a record of the volume of study drug 
suspension administered to and consumed by the subject.   
The study drugs will be inspected for damage, contamination, discoloration, or particulate matter 
before use. Any study drug that fails inspection will be quarantined at appropriate temperature 

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and labeled ‘Do Not Use’ until further notice. The Site PI or responsible person will immediately 
contact the DMID Product Support Team at DMIDProductSupportTeam@niaid.nih.gov and 
DMID Clinical Project Manager (CPM) for further instructions before administering any 
additional study drug. Based on the information collected, DMID and/or Entasis will determine 
whether the affected study drug can be used. If it cannot be used, the CTU will receive specific 
instructions on how to return it to DMID CMS or destroy it on site. Refer to the protocol-specific 
MOP for detailed information on the preparation and administration of zoliflodacin, placebo, and 
moxifloxacin. 
6.3 
Modification of the Study Intervention/ Investigational Products for 
a Subject 
Not applicable for single-dose study drug trial. See Study Halting Criteria, Section 9.5.1. 
6.3.1 
Overdose 
An overdose is defined as a dose greater than the high-dose level evaluated in the trial. All 
overdoses will be reported; if the overdose is associated with an AE, then the AE will also be 
reported. In the event of an overdose of zoliflodacin, the investigator will use clinical judgment 
in treating the overdose and contact the DMID Medical Monitor (MM). The investigator will 
refer to the relevant document(s) for detailed information regarding warnings, precautions, 
contraindications, AEs, and other significant data pertaining to zoliflodacin. Such documentation 
may include but not be limited to the IB. 
6.4 
Accountability Procedures for the Study Intervention/ 
Investigational Products   
The Site PI is responsible for the distribution and disposition of the study drugs and has ultimate 
responsibility for accountability. The Site PI may delegate this responsibility to the Site Research 
Pharmacist (RP). If delegated, the Site RP will be responsible for maintaining complete records 
and documentation of the study drugs’ receipt, accountability, dispensation, temperature 
monitoring, storage conditions, and final disposition. 
All study drugs, whether administered or not, will be documented on the appropriate study drug 
accountability record or dispensing log. Used and unused zoliflodacin sachets, placebo and 
moxifloxacin will be retained until monitored and released for disposition per DMID 
requirements. Details will be provided in the MOP.  
Upon completion of the trial and after the final monitoring visit, any remaining unused study 
drugs will either be returned or destroyed appropriately at the CTU as per DMID requirements 
and instructions that will be communicated to the CTU by the DMID CPM. 

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6.5 
Assessment of Subject Compliance with the Study Intervention/ 
Investigational Products  
Since each dose of zoliflodacin, placebo or moxifloxacin will be administered by site personnel, 
subject compliance is not anticipated to be an issue. Complete information regarding any partial 
or interrupted dosing will be documented. Subjects unable to ingest the full amount of the initial 
60-mL zoliflodacin or placebo suspension or the moxifloxacin tablet, or who vomit up to 5 h 
after dosing with zoliflodacin, placebo or moxifloxacin will be withdrawn from data analysis for 
the period when the event occurred, but will not be withdrawn from the trial and may complete 
the remaining treatment periods in the assigned dosing sequence. Subjects who do not ingest the 
content of the second 60-mL rinse will not be withdrawn from the trial. 
6.6 
Prior and Concomitant Medications/Treatments 
Medications include the following: prescription drugs, birth control hormonal preparations, non-
prescription medication, herbs, vitamins, nutritional supplements, and illicit and recreational 
substances.  
Medications taken before or after first dosing will be reported as Prior Medications or 
Concomitant Medications (ConMeds), respectively.  
Prior prescription medications will be recorded at Screening Visit and will not be taken for 30 
days before dosing or during the trial. Exceptions include: oral contraceptives; H1 
antihistamines; all medications approved for control of intraocular pressure including topical 
ophthalmic non-selective β-blockers, such as betaxolol, carteolol, levobunolol, metipranolol, and 
timolol; topical/ intranasal corticosteroids; NSAIDS; licensed influenza vaccine during the flu 
season, 7 days before the first dose or 7 days after the last dose. 
Zoliflodacin will not be co-administered in subjects taking medications that are strong (e.g., 
avasimibe, carbamazepine, phenytoin, rifampin, and St. John's wort) or moderate (e.g., efavirenz, 
bosentan, etravirine, modafinil, and nafcillin) inducers of CYP3A4. These medications will not 
be taken for 30 days before dosing and during the trial. 
Zoliflodacin will not be co-administered in subjects taking medications that pose a risk of TdP in 
association with prolonged QT/QTc (Appendix E and 17). These medications will not be taken 
for 30 days before dosing and during the trial.  
Moxifloxacin will not be co-administered in subjects taking multivalent cation-containing 
products including antacids, sucralfate, and multivitamins.  
A vaccine will not be received within 30 days before dosing or during the trial, except for 
licensed influenza vaccine during the flu season, which may be administered up to 7 days before 
the first dose or 7 days after the last dose.  

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Non-prescription medications, herbs, vitamins, and nutritional supplements will not be 
taken within 15 days before dosing and during the trial. Exceptions include: (1) nutritional 
supplements, juice, and herbal preparations or other foods or beverages that may affect various 
drug-metabolizing enzymes and transporters (e.g., grapefruit, grapefruit juice, grapefruit-
containing beverages, apple or orange juice, vegetables from the mustard green family [e.g., 
kale, broccoli, watercress, collard greens, kohlrabi, Brussels sprouts, mustard], and charbroiled 
meats) within 7 days before dosing; (2) St. John’s wart within 30 days before dosing and during 
the trial; and (3) vitamins and OTC medications taken for <48 h for the treatment of common 
symptoms (e.g., headache, indigestion, muscle pain) may be allowed if approved by the 
designated study clinician. 
Blood/blood products (RBCs, WBCs, platelets, and plasma) will not be donated within 60 days 
of dosing or received before Final Visit. 
Following dosing, each new ConMed and changes to existing medications will be recorded. 
Subjects will be required not to utilize non-study medications during the trial except those 
deemed necessary by the Site PI or sub-investigator. 
Any drug (e.g., non-prescription medications, herbal supplements, vitamins, or prescription 
medications) or vaccines or blood/blood products used by the subject during the trial will be 
recorded in the subject’s source documents and on the appropriate electronic case report form 
(eCRF), and the PI or authorized study clinician (listed on FDA Form 1572) will note whether 
the use was medically indicated and immediately necessary. Any use of medications not 
authorized by the study PI or authorized clinician will be recorded as a deviation.  
6.7 
Subject Restrictions 
6.7.1 
Physical Activity 
Subjects will be asked to refrain from rigorous physical activity 15 days before first dosing and 
during the trial.  
6.7.2 
Fluid and Food intake 
Subjects will be provided food and non-alcoholic beverages by the CTU during the dosing period 
of the trial. Subjects will fast for at least 8 h before dosing and 4 h after dosing. Water can be 
taken ad lib during those periods. For clinical laboratory blood draws, subjects will fast for at 
least 4 h. Meals during each dosing period will be standardized. Otherwise, there will be no 
restrictions regarding food and fluid intake.  

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6.7.3 
Alcohol, Marijuana and Illicit Drugs 
Alcohol will not be used at least 3 days before first study drug administration and during the 
trial. Marijuana and illicit drugs are exclusionary if detected before first dosing and prohibited 
during the trial. 
6.7.4 
Other Restrictions 
Smoking or use of tobacco or nicotine-containing products is prohibited within 30 days before 
first dosing and during the trial. Caffeinated beverages or foods containing caffeine are 
prohibited within 3 days before first dosing and during the trial. 

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7 
STUDY SCHEDULE 
The Schedule of Study Procedures and Evaluations is included as Appendix A. 
7.1 
Recruitment 
The subject population will be recruited from the local population surrounding the CTU, utilizing 
the CTU subject database and IRB-approved advertisements and social media. IRB-approved, 
prescreening questionnaires will be used to determine if subjects meet study requirements before 
scheduling screening visits. 
7.2 
Screening Visit (Day -21 to Day -2) 
The following will be done within 21 days before study drug dosing: 
• Obtain informed consent 
• Assign a study ID number to subjects who consent to participate 
• Review inclusion/exclusion criteria to confirm the subject is eligible for enrollment 
• Record demographics including age, gender, race, and ethnicity 
• Obtain contact information 
• Obtain MH   
• Review history of Prior Medications, including all taken within 30 days 
• Perform complete PE (except genital, rectal, and breast exams) by licensed clinician 
listed on Form FDA 1572 
• Obtain height and weight, and calculate BMI (wt [kg] / ht [m2]) 
• Take VS (supine systolic and diastolic BP, HR, respiratory rate, and oral temperature) 
• Obtain blood and urine samples for clinical laboratory tests  
• Obtain blood samples for viral serology  
• Obtain serum for β-HCG pregnancy test from all women  
• Obtain serum for FSH level from only post-menopausal women  
• Obtain urine sample for toxicology  
• Perform urine test for alcohol use 
• Obtain a 12-lead ECG with 10-sec rhythm strip 
• Counsel on the avoidance of pregnancy for women of childbearing potential 
• Counsel both male and females on the use of contraception 
• Counsel on the avoidance of alcohol, marijuana, illicit drugs, and prohibited medications 
• Counsel to avoid rigorous physical activity 

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Subjects who meet the eligibility criteria will be contacted by CTU personnel and asked to return 
to the CTU for Day -1 assessments and possible inpatient admission.  
Subjects who fail screening due to a medical condition or abnormal laboratory tests including 
pregnancy test and positive tests for HIV, HBV, and HCV will be informed of the findings and 
counseled to seek medical care for further evaluation and treatment. If subjects test positive for 
HIV antibody, HBsAg, and/or HCV antibody, they will be informed that test results may be 
reported to the local health authorities according to state or local law. Subjects who fail screening 
will not be re-screened unless they had an inter-current, short-term medical illness. Subjects who 
meet eligibility criteria but could not be enrolled within permissible window may be rescreened.  
7.3 
Check-in / Baseline (Day-1 in each dosing period)                                          
(Overall, Days -1, 8, 16, and 24 for dosing periods 1, 2, 3 and 4, respectively) 
Subjects meeting all inclusion and no exclusion criteria at Screening Visit will check into the 
CTU on Day -1 and the following procedures will be performed: 
• Review inclusion/exclusion criteria to confirm the subject remains eligible for enrollment 
• Update MH (Day -1 in Period 1 only) 
• Update Prior Medications  
• Perform abbreviated PE (Perform on each check-in. In Period 1, do not perform only if 
the screening PE was completed ≤7 days from dosing.)  
• Obtain VS 
• Obtain weight 
• Obtain blood and urine samples for clinical laboratory tests (collect on each check-in). In 
Period 1, do not collect if screening samples were collected ≤7 days from dosing.) 
• For all women, a serum β-HCG pregnancy test will be done, and negative results 
confirmed before dosing 
• Obtain urine sample for toxicology  
• Perform urine alcohol test for alcohol use 
• Obtain 12-lead standard ECG with 10-sec rhythm strip 
• Counsel to avoid rigorous physical activity 
• Review AEs/SAEs (Day -1 in Periods 2, 3, and 4, respectively, on study Days 8, 16, and 
24) 
 
Subjects who still meet the eligibility criteria on Day -1 before Period 1 will be admitted to the 
CTU on the same day.  
For subsequent dosing periods (2, 3, and 4), subjects will be admitted on Day -1.  Any new 
symptoms or abnormal assessments will be reviewed prior to dosing, and recorded as AEs.  

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Eligibility for continuation of dosing (dose 2, 3 and 4): 
Dosing for subsequent periods may proceed if: 
• A negative serum pregnancy test on Day -1 of each dosing period 
• Females of childbearing potential and males still agree to use acceptable contraception 
for the duration of the trial and for 30 days (females) or 90 days (males) after Final Visit 
• One of the following: 
o Subjects who did not experience any AE since the last dose administration  
o Any new or prior AEs were grade 1 or resolved, and in the opinion of the site 
principal investigator or appropriate sub-investigator, the AEs, individually or in 
cluster, do not pose a risk to the subject or would be likely to confound interpretation 
of the results. 
o Any new or prior AEs that were grade 2, should be either resolved or downgraded to 
Grade 1 and in the opinion of the site principal investigator or appropriate sub-
investigator, the AEs, individually or in cluster do not pose a risk to the subject or 
would be likely to confound interpretation of the results. 
o Any new or prior grade 3 AEs that were judged unrelated to study drug should be 
either resolved or downgraded to Grade 1 and in the opinion of the site principal 
investigator or appropriate sub-investigator, the AEs, individually or in cluster do not 
pose a risk to the subject or would be likely to confound interpretation of the results. 
If a subject has a Grade 3 AE judged related to study drug administration, this subject will be 
withdrawn from the study and will not receive subsequent dosing.  For all other AEs, dosing may 
be delayed up to 14 days in order to follow the AE for downgrading  or resolution according to 
the above.  If the subject does not meet the criteria for dosing specified above in 14 days, this 
subject will be withdrawn from the study and will not receive subsequent dosing. 
After admission to the CTU, the subject will fast at least 8 h before dosing but will have access 
to water during that period. 
7.4 
Inpatient period (Days 1-2 in each dosing period)                             
(Overall, Days 1-2, 9-10, 17-18, and 25-26 for dosing periods 1, 2, 3 and 4,  
respectively) 
7.4.1 
Randomization (Day 1, Dosing Period 1 only) 
A subject who continues to meet eligibility criteria by assessments performed before dosing will 
be enrolled in the trial and randomized on Day 1 of Dosing Period 1 to one of the 12 study 
treatment sequences. 
7.4.2 
Administration of Study Drug (Day 1 of each dosing period)                             
(Overall, Days 1, 9, 17, and 25 for dosing periods 1, 2, 3 and 4, respectively) 
Before Dosing:   
• Withhold breakfast but allow access to water   
• Review inclusion/exclusion criteria to confirm the subject remains eligible for dosing 

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• Update MH (Period 1) / AE review (Periods 2, 3, and 4) 
• Update Prior Medications /Document concomitant medications (ConMeds) (Periods 2, 3, 
and 4) 
• Obtain VS within 60 min before dosing 
• Perform symptom-directed PE, if applicable 
• Obtain 12-lead standard ECG with 10-sec rhythm strip within 60 min before dosing 
• May insert IV catheter for blood collection into a forearm vein 
• Obtain blood (plasma) PK sample within 30 min before dosing 
• Initiate 12-lead Holter ECG at least 1 h before dosing 
o Subjects will remain in a supine position at rest for at least 10 min before and 5 min 
after each of the following timepoints: 45, 30, and 15 min before dosing.  
o If timepoints for ECG extraction coincide with 12-lead standard ECGs, VS 
assessments, and PK blood draws, these procedures will be performed in this order: 
12-lead Holter ECG, 12-lead standard ECG, VS, and PK blood draw.   
Dosing:  
• Administer a single oral dose of the study drug with water, as described in Section 6.2, 
according to the randomization schedule for each treatment sequence and period 
• Withhold food until 4 h after dosing but allow access to water 
After Dosing: 
• Continue 12-lead Holter ECG: 
o Subjects will remain in a supine position at rest for at least 10 min before and 5 min 
after each of the following timepoints: 0.5, 1, 2, 3, 4, 6, 8, and 12 h after dosing.  
o If timepoints for ECG extraction coincide with 12-lead standard ECGs, VS 
assessments, and PK blood draws, these procedures will be performed in this order: 
12-lead Holter ECG, 12-lead standard ECG, VS, and PK blood draw.  
• Obtain 12-lead standard ECG with 10-sec rhythm strip at 1 h (±10 min), 2 h (±10 min), 
and 4 h (±10 min) after dosing 
• Obtain VS at 1 h (±10 min), 2 h (±10 min), and 4 h (±10 min) after dosing; more frequent 
monitoring will be at the PI’s discretion based on subject’s clinical status 
• Obtain blood (plasma) PK samples at 0.5 h (±5 min), 1 h (±5 min), 2 h (±5 min), 3 h (±10 
min), 4 h (±10 min), 6 h (±10 min), 8 h (±15 min), and 12 h (±15 min) after dosing 
• Perform symptom-directed PE, if applicable 
• Perform AE and SAE assessments 
• Document ConMeds 

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7.4.3 
Inpatient Follow-up (Day 2 of each dosing period)                                            
(Overall, Days 2, 10, 18, and 26 for dosing periods 1, 2 3 and 4, respectively) 
• Continue 12-lead Holter ECG: 
o 
Subjects will remain in a supine position at rest for at least 10 min before and 5 min 
after the following timepoint: 24 h (±2 h) after dosing.  
o 
If the timepoint for ECG extraction coincides with 12-lead standard ECG, VS 
assessment, and PK blood draw, these procedures will be performed in this order: 
12-lead Holter ECG, 12-lead standard ECG, VS, and PK blood draw.  
• Obtain VS 
• Perform symptom-directed PE, if applicable 
• Perform AE and SAE assessments 
• Obtain clinical labs before discharge 
• Obtain blood (plasma) PK samples at 24 h (±2 h) after dosing 
• Document ConMeds 
• Obtain 12-lead standard ECG at 24 h (± 2 h) after dosing (before discharge on Day 2) 
• Remove 12-lead Holter recorder on Day 2 at 24 h after dosing after last recording 
• Remind women of childbearing potential on the avoidance of pregnancy 
• Remind male and female subjects on the use of appropriate contraception 
• Remind male subjects to refrain from sperm donation  
• Remind subjects to abstain from prohibited medications, alcohol, marijuana, and illegal 
drugs 
• Remind subjects to abstain from rigorous physical activity 
• Instruct on the next scheduled visit 
• Discharge subject from the CTU after review of clinical laboratory tests, ECGs, and other 
assessments by PI or authorized clinician 
7.5 
Outpatient period (Days 3-7 for each dosing period                           
(Overall, Days 3-7, 11-15, 19-23 and 27-31 for dosing periods 1, 2, 3 and 4, 
respectively) 
This will be a 5-day outpatient period between completion of treatment in the previous period 
and re-admission to the CTU for next dosing period except after Dose 4. Subjects will report to 
the CTU any new AEs/SAEs they have experienced, and medications they have used during the 
outpatient periods.  
7.6 
Final Visit (Day 32, 8 ±2 days after last dose in Dosing Period 4) 
• Obtain VS 
• Perform complete PE  

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• Obtain weight 
• Collect blood for serum β-HCG pregnancy test in all females 
• Perform AE and SAE assessments 
• Update ConMeds 
• Obtain 12-lead standard ECG with 10-sec rhythm strip 
• Obtain blood and urine for clinical laboratory tests  
• Remind male and female subjects on the avoidance of pregnancy 
• Remind female subjects to use appropriate contraception for 30 days after Final Visit 
• Remind male subjects to use appropriate contraception and refrain from sperm donation 
for 90 days after Final Visit 
• Discharge subject from the trial 
7.7 
Early Termination Visit (if needed) 
• Obtain VS 
• Perform complete PE  
• Perform AE and SAE assessments 
• Update ConMeds 
• Obtain 12-lead standard ECG with 10-sec rhythm strip 
• Obtain blood and urine for clinical laboratory tests  
• Collect blood for serum β-HCG pregnancy test in all females  
• Obtain blood PK sample if ET occurs within 24 h of dosing 
• Remind male and female subjects on the avoidance of pregnancy 
• Remind female subjects to use appropriate contraception for 30 days after ET 
• Remind male subjects to use appropriate contraception and refrain from sperm donation 
for 90 days after ET 
7.8 
Unscheduled Visit (if needed) 
A subject may return to the clinic for an unscheduled visit at any time. The following activities at 
a minimum will be performed:  
• Obtain VS 
• Perform symptom-directed PE, if applicable 
• Update ConMeds 
• Perform AE and SAE assessments 
• Obtain blood and/or urine for clinical laboratory tests, if applicable 
• Obtain 12-lead standard ECG with 10-sec rhythm strip as needed for the evaluation of an 
AE 
• Remind male and female subjects on the avoidance of pregnancy 

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8 
STUDY PROCEDURES/EVALUATIONS 
8.1 
Clinical Procedures/Evaluations 
8.1.1 
Informed Consent 
The ICF will be approved by the reviewing IRB/EC and executed before performing any study-
related activities. 
Informed consent will be obtained for all subjects participating in the trial before performing any 
screening assessments. Subjects may withdraw consent at any time. Participation in the trial may 
be terminated at any time without the subject’s consent as determined by the investigator. 
8.1.2 
Demographics 
Demographic information (date of birth, gender, ethnicity, and race) will be recorded on the 
subject’s source documents and eCRF at Screening Visit. Name, address, phone number, and 
emergency contact information will be documented in the source documents only. 
8.1.3 
Inclusion/Exclusion Criteria 
Eligibility screening of healthy subjects will be completed within 21 days before first study drug 
dosing and will be documented on the subject’s source documents and eCRF. Confirmation of 
eligibility will be performed before dosing on Day 1 of each dosing period. 
Screening failures and the reason for failure to meet the study eligibility requirements will be 
documented in the source documents and entered into the study database. 
8.1.4 
Medical History 
For subjects enrolled in the trial, the MH will be obtained by direct interview of the subject and 
recorded on the subject’s source document and eCRF. The MH will capture the subject’s current 
disease processes, past disease processes, history of hospitalization, history of surgery, allergies, 
and prior medications (taken 30 days before dosing). Subjects will be queried regarding a history 
of significant medical disorders of the head, eyes, ears, nose, throat (HEENT), mouth, skin, and 
the cardiovascular, gastrointestinal, renal, urological, nervous, hematological, lymphatic, 
endocrine, musculoskeletal, and genital/reproductive systems. A history of cardiovascular 
disorders, including MI and angina, cardiac arrhythmias including LQTS, syncope due to cardiac 
arrhythmias or unexplained, psychiatric illness, and substance abuse will be specifically 
solicited. Family history especially of sudden death will be obtained. The MH will be obtained at 
Screening Visit and updated upon admission to the CTU and before dosing on Day 1 in Period 1. 
After first study drug dosing, any worsening of pre-dosing MH or new symptoms will be 
evaluated and reported as AEs. 

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8.1.5 
Physical Examination 
A complete PE – except genital, breast, and rectal exams – will be performed at Screening Visit 
and Final Visit (Day 8 ±2 after Dosing Period 4), or ET Visit, and will assess general 
appearance, HEENT, heart, lungs, abdomen, skin, musculoskeletal system, and lymph nodes, 
and include an abbreviated neurological exam. 
An abbreviated PE will be performed in each dosing period on Day -1 in each inpatient period. If 
a complete PE was performed ≤7 days before first dosing, then an abbreviated PE will not be 
performed on Day -1 of the Dosing Period 1. An abbreviated PE differs from a complete PE in 
that the abdomen and neurological system are not evaluated.  
A symptom-directed PE will be performed in each dosing period before dosing on Day 1 and 
after dosing on Days 1 and 2, to evaluate new symptoms or treatment-emergent AEs, 
respectively.  
Height and weight will be measured, and BMI calculated, at Screening Visit; only weight will be 
measured on Day -1 of each dosing period and at Final Visit (Day 8 ±2 after Dose 4) or ET. 
Refer to the protocol-specific MOP for further details. The findings of each examination will be 
recorded on the subject’s source documents and eCRF. Any new findings on examination or 
worsening of existing conditions after dosing are to be reported as AEs.  
8.1.6 
Vital Signs 
VS including resting (measured after supine for at least 5 min) systolic and diastolic BP, HR, 
respiratory rate, and oral temperature will be measured at Screening Visit, on Day -1, before and 
after dosing on Day 1, and on Day 2 of each of the four dosing periods, and at Final Visit after 
Dose 4, or ET. On Day 1 of each dosing period, VS will be measured approximately 1 h before 
dosing, and 1 h (±10 min), 2 h (±10 min), and 4 h (±10 min) after dosing. On Day 2, VS will be 
measured at 24 h (±2 h) after dosing. Acceptable ranges are shown in Appendix B. 
VS that are considered aberrant due to an error in measurement may be repeated. At Screening 
and after dosing, an abnormal VS measurement may be repeated twice more at rest, within 5 min 
of each other. If the second measurement is abnormal, it will be reported at the highest grade of 
the two measurements and the subject will be excluded (if at Screening Visit) or the event 
reported as an AE (if after dosing). If the second measurement is normal, a third measurement 
will be taken at least after 5 min at rest. If the third measurement is still normal, the subject is 
eligible (if at Screening Visit) or there is no AE (if after dosing); if it is abnormal, the subject 
will be excluded (if at Screening Visit) or an AE will be reported at the highest assessed grade 
(between first and third measurements) and graded for severity per Appendix C (if after dosing). 

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8.1.7 
12-lead Standard Electrocardiogram (ECG) 
A 12-lead standard ECG and 10-sec rhythm strip will be obtained at Screening Visit and on Day 
-1, Day 1 before and after dosing, Day 2 (24 ±2 h after dosing) in each treatment period, and 
Final Visit after Dose 4, or ET. On Day 1 of each dosing period, a 12-lead standard ECG and 10-
sec rhythm strip will be recorded within 1 h before dosing and 1 h (±10 min), 2 h (±10 min), and 
4 h (±10 min) after dosing. The ECGs will be reviewed by the PI or a designated clinician (listed 
on FDA Form 1572). ECGs will be performed after the subject rests quietly in a supine position 
for at least 5 min. To be eligible for participation, the QT/QTcF interval must be normal and 
there must be no clinically significant ECG abnormalities. If a question regarding ECG 
interpretation arises, the study investigators will have the ECG reviewed by a cardiologist. 
8.2 
Laboratory Evaluations 
Venipuncture schedule and blood volumes are shown in Appendix A and Appendix D. The 
blood volume for HEM and CHEM tests will be approximately 9 mL per sample (Appendix D). 
8.2.1 
Clinical Laboratory Evaluations (Hematology, Chemistry, and Urinalysis) 
Blood and urine samples for clinical laboratory tests will be collected at Screening Visit, on Day 
-1 of each dosing period (with the exception in the first dosing period if screening clinical labs 
were collected ≤7 days of Dose 1), on Day 2 of each dosing period, and Final Visit after Dose 4, 
or ET. Subjects must be fasting for > 4 h before any blood draw for clinical laboratory 
assessments. These tests will include: 
• HEM: Hgb, Hct, RBC count, platelet count, and WBC count with absolute differential 
count 
• CHEM: serum creatinine, with estimation of GFR, blood urea nitrogen (BUN), glucose 
(fasting at least 4 h), total bilirubin, direct bilirubin, alanine aminotransferase (ALT), 
aspartate aminotransferase (AST), alkaline phosphatase (AP), total protein, albumin, 
sodium, chloride, potassium, total carbon dioxide (CO2), magnesium 
• UA: Routine dipstick testing of clean-catch urine for blood, protein, and glucose; other 
coincidental tests will be recorded   
Clinical laboratory tests at Screening Visit and Day -1 of Treatment Period 1 should be in the 
normal reference range with acceptable exceptions, as shown below and in Appendix B: 
Elevated bilirubin due to documented Gilbert syndrome that is Grade 1 is allowable, but Grade 2 
or higher is exclusionary. [To document a subject has Gilbert syndrome, a diagnosis from the 
medical record must be provided or the PI may make a ‘presumptive’ diagnosis of Gilbert 
syndrome in subjects with unconjugated hyperbilirubinemia on repeated testing (at least two 
samples separated in time) who have otherwise normal serum ALT, AST, and AP concentrations, 
and a normal CBC.]  

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Elevated serum glucose, sodium, potassium, bicarbonate (CO2), total protein, and AP values that 
reach toxicity Grade 1 values are allowable. 
Serum chloride and albumin above the upper limit of normal (ULN) is allowable. 
If UA by dipstick is not within acceptable ranges per Appendix B for blood, protein, or glucose, a 
complete UA with microscopic evaluation will be performed and the results will supersede those 
of the dipstick. Menstruating females failing inclusion criteria due to a positive urine dipstick or 
microscopic UA may be retested after cessation of menses. Do not exclude subjects with ≤6 
RBC/HPF. 
Laboratory values that are outside the range of eligibility but are thought to be due to an acute 
condition or due to collection or laboratory error may be repeated once.   
Laboratory values will be transferred to AdvantageEDCSM.  
Abnormal safety laboratory values after dosing will be graded per the Toxicity Table, Appendix 
C. Abnormal values within the acceptable range per Appendix B noted at screening or baseline 
will only be considered AEs if they deteriorate after study drug dosing. 
8.2.2 
Viral Serology Testing 
Serological testing for HIV antibody, HBsAg, and HCV antibody will be performed at Screening 
Visit. These tests must be negative for study eligibility. In cases where a false-positive result is 
suspected, confirmatory testing (e.g., polymerase chain reaction) may be performed. 
8.2.3 
Pregnancy Testing 
In all women, a serum β-HCG level will be measured at Screening Visit and on Day -1 of each 
dosing period; results must be negative for entry into each dosing period of the trial. Serum 
pregnancy test will also be performed at Final Visit, or ET. 
8.2.4 
Serum FSH Testing 
A serum FSH level for confirmation of post-menopausal status in female subjects will be 
measured at Screening Visit only.   
8.2.5 
Urine Toxicology Screening 
A urine toxicology screen will be performed at Screening Visit and on Day -1 to detect the 
presence of amphetamines, cocaine (and metabolite), barbiturates, benzodiazepines, opiates, 
MDMA, methadone, tetrahydrocannabinol, methamphetamines, tricyclic antidepressants 
(TCAs), and phencyclidine. Results must be negative for study eligibility. Urine creatinine will 
be measured as part of the profile to assess quality of collected sample.  

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8.2.6 
Urine Alcohol Testing  
Detection of recent alcohol consumption in urine will be performed at Screening Visit and on 
Day -1 in each treatment period. Results must be negative for study eligibility.  
8.3 
Special Procedures and Assays 
8.3.1 
Continuous 24-h ECG Monitoring (Holter ECG Monitoring) 
ECGs for assessment of the primary endpoint will be obtained digitally during all four dosing 
periods using a continuous 12-lead Holter ECG digital recorder system. The continuous ECG 
recording will be started approximately 1 h before dosing on Day 1 of all dosing periods and will 
continue until 24 h after dosing (i.e., in the morning of Day 2). Three replicate ECG recordings 
will be extracted at the following timepoints in each dosing period: 45, 30, and 15 min before 
dosing (baseline) and 0.5, 1, 2, 3, 4, 6, 8, 12, and 24 h after dosing for a total of 12 timepoints per 
subject per dosing period. The subject will be in a supine position for at least 10 min before 
recording an ECG and 5 min after the nominal timepoint for the ECG extractions from the Holter 
monitors. These timepoints will correspond to PK blood sampling for each dosing period, but 
ECGs will be obtained before the measurement of VS and collection of blood samples, to avoid 
the effect of stress on the ECG. The mean of pre-dose ECGs extracted on Day 1 of each dosing 
period will be used as baseline reference for results obtained after dosing. 
ECGs will be extracted from the Holter recordings over a 5-min period and used to measure 
ECG intervals. Digital QT, QTcF, PR, QRS, and RR intervals and HR will be determined by the 
Core ECG Lab. Digital ECGs will be read in accordance with the principles outlined in ICH E14 
Guidance for TQT studies.16  
The 12-lead Holter and ECG equipment will be supplied and supported by the VACR Phase 1 
CTU. All ECG data will be collected using a continuous 12-lead ECG digital recorder. The 
continuous 12-lead digital ECG data will be stored onto secure digital memory cards. ECGs to be 
analyzed will be collected at pre-determined timepoints (as indicated above and in Appendix A: 
Schedule of Study Procedures and Evaluations) and will be read centrally by IQVIA Cardiac 
Safety Services. 
The following principles will be followed in the IQVIA Cardiac Safety Services core laboratory: 
• Each extracted ECG will be analyzed by the cardiologist who will be blinded to subject 
identifiers, time, and treatment.  
• Electrocardiograms will be analyzed for PR, QRS, RR, and QT intervals using semi-
automated measurements and for morphological abnormalities including T and U waves 
manually by experienced cardiologists.  
• A limited number of skilled readers will be employed in the over-read process to control 
for variability in interpretation.  
• All ECGs from a single subject will be read together by the same reader.  

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• Consistent leads will be used for estimation of ECG intervals to the extent possible, with 
lead II as the primary lead. If technical issues prevent measurement in lead II, the QT will 
be measured in an alternative lead as per the core laboratory procedure.  
The following sections provide a brief description of ECG analysis methods utilized by the core 
laboratory. 
8.3.1.1 
TQT and ECG Extraction Technique 
Three replicate ECG recordings will be extracted at the indicated timepoints from each of the 
four dosing periods. For each timepoint, ECG extraction will typically be from the last 5 min of 
the period (at least 10 min) when the subject is maintained in a supine quiet position before the 
nominal timepoint (i.e., 25-30 min for the 0.5-h post-dosing timepoint and 55-60 min for the 1-h 
post-dosing timepoint). 
The ECG Core Lab will use Antares software to extract ECGs from the continuous 24-h Holter 
recordings after evaluating the complexes for sinus origin, heart rate stability criteria and the 
smallest noise score. During protocol-specified ECG extraction windows, three 10-sec digital 12-
lead ECG tracings will be extracted from continuous Holter recordings. The mean QT, QTcF, 
and RR values from each extracted replicate will be calculated, and the average of all mean 
values of primary ECG parameters from all replicate ECGs from a nominal timepoint will be 
used as the subject’s reportable value at that timepoint. 
The IQVIA Cardiac Safety Services will submit extracted ECG files in FDA-compliant HL 7 
XML to the FDA ECG warehouse. 
8.3.1.2 
T-wave Morphology Analysis and Measurement of PR and QRS Intervals 
Categorical T-wave morphology analysis will be performed manually in the three ECG replicates 
at each timepoint. Each fiducial point (onset of P-wave, onset of Q-wave, offset of S-wave, and 
offset of T-wave) will be electronically marked. 
The original ECG waveform (Table 2) and such annotations will be saved separately at the 
conclusion of the trial in Extensible Markup Language (XML) format for independent review 
and upload to the FDA ECG Warehouse. 
 
 

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Table 2: T-wave Morphology Categories (Assessed Manually) 
In addition to the T-wave categorical analysis, the presence of abnormal U waves will be noted. 
However, discrete U waves will be excluded from QT/QTc interval measurements. 
8.3.2 
Pharmacokinetics  
8.3.2.1 
Assay for Zoliflodacin 
Blood (plasma) samples for assay of zoliflodacin PK will be collected in K2EDTA tubes at the 
following study days and timepoints: On Day 1 within 30 min before dosing, and 0.5 h (±5 min), 
1 h (±5 min), 2 h (±5 min), 3 h (±10 min), 4 h (±10 min), 6 h (±10 min), 8 h (±15 min), and 12 h 
(±15 min) after dosing; and on Day 2 at 24 h (± 2 h) after dosing, or ET if occurs within 24 h of 
dosing. 
Blood samples will be obtained promptly after each corresponding Holter recording and 12-lead 
standard ECG timepoint and VS assessment. 
Sample collections will be scheduled for the nominal timepoint and actual collection times 
recorded in source documents.  
Plasma concentrations of zoliflodacin will be determined using a validated HPLC-MS/MS 
method. 
8.3.2.2 
Assays for Placebo and Moxifloxacin Samples  
Blood samples obtained during placebo dosing periods will not be analyzed. Blood samples 
obtained during moxifloxacin dosing periods will be stored and analyzed only for-cause.  
 
Category 
Description 
Normal T-wave  
Any T-wave not meeting any criterion below 
Flat T-waves  
T amplitude <1 mm (either positive or negative) including flat 
isoelectric line 
Notched T-wave (+)  Presence of notch(es) of at least 0.05 millivolt (mV) amplitude on 
ascending or descending arm of the positive T-wave 
 
Biphasic 
 
T-wave that contains a second component with an opposite phase that 
is at least 0.1 mV deep (both positive and negative/positive and 
polyphasic T-waves included) 
Normal T-wave (-) 
T amplitude that is negative, without biphasic T-wave or notches 
Notched T-wave (-)  
Presence of notch(es) of at least 0.05 mV amplitude on descending or 
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8.3.3 
Specimen Preparation, Handling, and Shipping 
8.3.3.1 
Instructions for Specimens Preparation, Handling, and Storage 
Details regarding the specimen preparation, handling, and storage are described in the protocol-
specific MOP. 
Blood samples left after all routine clinical laboratory testing and pharmacokinetic assays are 
completed will not be stored indefinitely or used for purposes other than those described. No 
genetic testing will be done on blood samples collected.  
8.3.3.2 
Specimen Shipment 
Specimen shipment will occur at intervals during the trial following all applicable International 
Air Transport Association (IATA) requirements and according to the specifics for storage 
temperature and documentation as detailed in the central clinical laboratory manual and protocol-
specific MOP, as appropriate. 
All specimens for clinical screening and safety laboratory evaluations will be transported from 
the CTU to the local clinical laboratory. 
Plasma samples for bioanalytical assays will be shipped from the CTU to DMID-CMS at: 
Fisher BioServices 
c/o DMID Clinical Materials Services (CMS) 
20439 Seneca Meadows Parkway 
Germantown, MD 20876 
Phone: 240-477-1350  
Fax: 240-477-1360 
Email: DMID.CMS@thermofisher.com  
Plasma samples will then be provided by DMID-CMS to the bioanalytical lab, KCAS Inc., at:  
KCAS Bioanalytical Services  
c/o Marsha Luna, Senior Manager, PBI 
12400 Shawnee Mission Parkway 
Shawnee, KS 66216 
Office: 913-248-3042  
Email: marsha.luna@kcasbio.com  
 
 
 

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9 
ASSESSMENT OF SAFETY 
Regulatory requirements including FDA regulations and ICH Guidelines for GCP set forth safety 
monitoring and reporting responsibilities of sponsors and investigators to ensure the safety and 
protection of human subjects participating in clinical trials. 
Responsibilities: 
Investigators participating in this clinical trial are responsible for and will: 
• Evaluate subject safety including assessment of AEs for seriousness, severity, and 
causality (relatedness to study drug); 
• Notify DMID of SAEs within 24 h of site awareness; 
• Provide detailed written reports, including necessary documentation requested by DMID 
or IRB/EC, promptly following immediate initial reports; 
• Inform the IRB/EC of SAEs and AEs as required by applicable regulatory requirements. 
9.1 
Specification of Safety Parameters 
Safety will be assessed by the timing, frequency, causality, and severity of: 
1. Treatment-emergent SAEs occurring from time of first dose in dosing period 1 through 
Final Visit (Day 8 ±2 after last dose in dosing period 4), or ET; 
2. Clinical laboratory AEs occurring from time of first dose in dosing period 1 through Final 
Visit (Day 8 ±2 after last dose in dosing period 4), or ET; 
3. Non-serious, unsolicited treatment-emergent AEs occurring from time of first dose in 
dosing period 1 through Final Visit (Day 8 ±2 after last dose in dosing period 4), or ET. 
9.2 
Methods and Timing for Assessing, Recording, and Analyzing Safety 
Parameters 
9.2.1 
Adverse Events 
Definitions 
ICH E6 defines an AE as any untoward medical occurrence in a subject or clinical investigation 
subject who was administered a pharmaceutical product, regardless of its causal relationship to 
the product. An AE can therefore be any unfavorable and unintended sign (including an abnormal 
laboratory finding), symptom, or disease temporally associated with use of the product. The 
occurrence of an AE may come to the attention of study personnel during study visits and 
interviews, or by a subject presenting for medical care. 

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The FDA defines an AE as any untoward medical occurrence associated with the use of a drug in 
humans, whether it is considered drug-related or not. 
Any medical condition that is present at the time that the subject is screened will be considered a 
baseline finding and not reported as an AE. However, if the condition increases in severity or 
frequency at any time during the trial, it will be recorded as an AE. 
All AEs will be graded for severity and relationship to the study drug. 
9.2.1.1 
Severity of Events 
Intensity of AEs will be graded as follows, unless otherwise specified in Appendix C: 
Mild: Require minimal or no treatment; do not interfere with the subject’s daily activities. 
Moderate: Result in a low level of inconvenience or concern with therapeutic measures; may 
cause some interference with functioning. 
Severe: Interrupt a subject’s usual daily activity and may require systemic drug therapy or other 
treatment; are usually incapacitating. 
9.2.1.1.1 Relationship to Study Products 
AEs and SAEs will be assessed by the investigator to determine relationship to the study drug, 
using the following two terms. In a clinical trial, the study drug must always be suspect.   
• Related: There is a reasonable possibility that the study drug caused the AE/SAE 
• Not Related: There is not a reasonable possibility that the study drug caused the AE/SAE 
The investigator will provide an assessment of association or relationship of each AE/SAE to the 
study drug based on: 
• Temporal relationship of the AE/SAE to study drug dosing; 
• Whether an alternative etiology has been identified; 
• Biological plausibility; 
• Existing therapy and/or ConMeds. 
9.2.1.2 
Reporting Adverse Events 
AEs will be captured on the appropriate subject’s source document and eCRF. Information 
collected for AEs includes event description, time of onset, investigator assessment of severity 
and relationship to the study drug, date of resolution of the event, seriousness, and outcome. 
All AEs will be documented from the time of study drug dosing through Final Visit. AEs will be 
followed to resolution or until considered stable in the clinical judgment of the study investigator. 
Evaluation of AEs may require unscheduled visits and clinical and laboratory investigations, 
according to the clinical judgment of the Site PI and study physicians. 

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9.2.2 
Serious Adverse Events 
An SAE is any AE that meets at least one of the following criteria: 
• Death; 
• Life-threatening AE*; 
• Inpatient hospitalization or prolongation of existing hospitalization; 
• Persistent or significant disability or incapacity, or substantial disruption of the ability to 
conduct normal life function; 
• Congenital anomaly/birth defect; 
• Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered an SAE when, based upon appropriate medical 
judgment, they may jeopardize the 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. 
*An AE is considered “life-threatening” if, in the view of either the investigator or DMID, 
its occurrence places the subject at immediate risk of death. It does not include an AE that, 
had it occurred in a more severe form, might have caused death. 
SAEs will be: 
• Assessed for severity and relationship to study product and alternate etiology by an 
authorized study physician (listed on FDA Form 1572); 
• Recorded on the appropriate SAE data collection form and eCRF; 
• Followed through resolution; 
• Reviewed and evaluated by an ISM (as deemed necessary), the SMC (periodic review 
unless related), DMID, and the IRB. 
9.2.3 
Procedures to be Followed in the Event of Abnormal Laboratory Test Values or 
Abnormal Clinical Findings 
A licensed study clinician (listed on FDA Form 1572) will determine seriousness, severity, and 
causality; provide a medical evaluation of AEs; and classify AEs based upon medical judgment. 
Abnormal laboratory values or clinical findings for all enrolled subjects after dosing will be 
assessed using the toxicity scales in Appendix C. Abnormal values and findings noted at 
screening or baseline will only be considered AEs if they deteriorate after study drug dosing. For 
abnormalities noted from the time of first study drug dosing, any Grade 1 or higher laboratory 
abnormality listed on the toxicity table in Appendix C will be entered in the database as an AE. 
Safety laboratory results that are abnormal according to the local laboratory reference range, but 
not considered Grade 1 abnormalities, will be evaluated by the CTU clinician and reported as 
Grade 1 abnormalities if clinically significant. If not clinically significant, these will not be 
considered laboratory AEs and thus will not be graded but will be recorded in the source 
document and followed-up clinically at the discretion of the CTU clinician. Abnormal laboratory 

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values, performed as part of HEM, CHEM, or UA but not listed in this toxicity table will be 
evaluated by the study clinicians, recorded in the source document and, if clinically significant, 
considered AEs and graded according to the criteria in Section 9.2.1. 
Protocol-specific laboratory normal range values in effect at the time of protocol submission are 
included in Appendix B. Changes in these ranges during the trial will be handled as 
administrative protocol amendments and toxicity grades will be calculated using the new ranges 
from the effective date of the change. A regular protocol amendment will be submitted for these 
changes if a revised laboratory normal range value meets criteria for Grade 2 toxicity. 
Gross blood in urine that is confirmed due to menses is not an AE (but is for all other reasons). 
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 AE is defined as the return to pretreatment status or stabilization of the condition 
with the expectation that it will remain chronic. 
All SAEs will be: 
• Recorded on the appropriate SAE report form and sent to DMID Pharmacovigilance 
Group (PVG) 
• Entered into the appropriate subject source document and eCRF in AdvantageEDCSM 
• Reported to the Site ISM and the IRB 
• Reviewed and followed to resolution or stability by an authorized study physician (listed 
on FDA Form 1572) 
• Collected on each subject until Final Visit or ET Visit 
Any AE that meets a protocol-defined serious criterion will be submitted immediately (within 
24 h of site awareness) on an SAE report form to DMID PVG: 
 
DMID Pharmacovigilance Group 
Clinical Research Operations and Management Support (CROMS) 
6500 Rock Spring Drive, Suite 650 
Bethesda, MD 20817, USA 
SAE Hot Line: 1-800-537-9979 (US) or 1-301-897-1709 (outside US) 
SAE FAX Number: 1-800-275-7619 (US) or 1-301-897-1710 (outside US) 
SAE Email Address: PVG@dmidcroms.com 
In addition to the SAE report form, selected SAE data fields will also be entered into 
AdvantageEDCSM. Please see the protocol-specific MOP for details regarding this procedure. 

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Other supporting documentation of the SAE may be requested by DMID PVG and will be 
provided as soon as possible. 
The CTU will copy the ISM on SAE reports provided to the DMID PVG. The DMID Medical 
Monitor (MM) and DMID CPM will be notified of the SAE by the DMID PVG. The DMID MM 
will review and assess the SAE for regulatory reporting and potential impact on subject safety and 
protocol conduct. 
At any time after completion of the trial, if the Site PI or appropriate sub-investigator becomes 
aware of an SAE that is suspected to be related to study drug, the Site PI or appropriate sub-
investigator will report the event to the DMID PVG. 
9.3.2 
Regulatory Reporting for Studies Conducted Under DMID-Sponsored IND 
Following notification from the investigator, DMID, the Investigational New Drug (IND) 
sponsor, will report any suspected AE that is both serious and unexpected. DMID will report an 
AE as a suspected AE only if there is evidence to suggest a causal relationship between the drug 
and the AE. DMID will notify FDA and all participating investigators (i.e., all investigators to 
whom DMID is providing drug under its IND(s) or under any PI’s IND(s)) 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 DMID determines that the information qualifies for 
reporting as specified in 21 CFR Part 312.32. DMID will also notify FDA of any unexpected fatal 
AE or suspected life-threatening AE as soon as possible, but in no case later than 7 calendar days 
after DMID’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 the study drug will be reported to the FDA at 
least annually in a summary format. 
9.3.3 
Other Adverse Events (if applicable) 
9.3.3.1 
Reporting of Overdose 
An overdose is defined as a dose greater than the high-dose level evaluated in the trial as 
described in Section 6.3.1 of the protocol. All overdoses will be reported; if the overdose is 
associated with an AE, then the AE will also be reported. In the event of an overdose of study 
drug, the investigator will use clinical judgment in treating the overdose and contact the DMID 
MM. The investigator will refer to the relevant document(s) for detailed information regarding 
warnings, precautions, contraindications, AEs, and other significant data pertaining to 
zoliflodacin. Such documentation may include but not be limited to the IB. 

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9.3.4 
Reporting of Pregnancy 
Pregnancies that occur in female subjects during the trial will be reported via AdvantageEDCSM 
on a pregnancy report form. With the subject’s permission, all protocol-required venous blood 
samples will be obtained, and the subject will continue to be followed for safety until Final Visit. 
Efforts will be made to follow all pregnancies reported during the trial to pregnancy outcome, as 
described in the protocol-specific MOP (e.g., delivery, spontaneous abortion, or therapeutic 
abortion), pending the subject’s permission. 
A female subject who participates in the trial and becomes pregnant will be asked to inform study 
personnel of a pregnancy occurring 30 days after Final Visit. A male subject who participates in 
the trial and whose female partner becomes pregnant from the time of first dosing to 90 days after 
Final Visit will be asked to inform study personnel of the pregnancy. For all reported pregnancies, 
subjects will be asked to provide pregnancy outcome upon delivery or pregnancy termination to 
the CTU.  
9.4 
Type and Duration of Follow-up of Subjects after Adverse Events 
Treatment-emergent SAEs and AEs will be followed until resolution or until considered stable in 
the clinical judgment of the investigator. Resolution of an AE is defined as the return to 
pretreatment status or stabilization of the condition in the clinical judgment of the study 
investigator, with the expectation that it will remain chronic. 
Follow-up procedures, evaluations, and outcomes will be recorded in the subject's source 
document and eCRF. 
9.5 
Halting Rules 
9.5.1 
Study Halting Criteria 
9.5.1.1 
Halting Criteria for Zoliflodacin and Placebo Periods 
Study dosing of zoliflodacin or placebo will be temporary halted, and a review of available safety 
data will be conducted by the SMC if any of the following occur after dosing:  
1. Death of a subject following dosing regardless of relatedness to the study drug; 
2. One subject with a non-fatal SAE that is considered related to the study drug by the study PI 
or authorized physician (listed in FDA Form 1572); 
3. Two or more subjects experience a QTcF >500 msec or ≥60 msec above baseline, based on 
readings from at least two standard 12-lead ECGs after study drug administration; 
4. Two or more subjects with a Grade 3 systemic AE coded in the same organ system that is 
considered related to the study drug by the study PI or authorized clinician; 
Note: VS abnormalities will be considered part of a systemic disorder or an organ-specific condition, as described 
in Appendix C: Adverse Events Toxicity Grading Criteria 

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, in order to be included among the Study Halting Criteria.  
5. One subject experiences a symptomatic ventricular tachyarrhythmia with or without QTcF 
prolongation after study drug administration; 
6. An overall pattern of symptomatic, clinical non-serious events that may appear minor in terms 
of individual events, but that may collectively represent a serious potential safety concern and 
pose a risk to continuing a subject in the trial. 
9.5.1.2 
Halting Criteria for Moxifloxacin Periods 
Study dosing of moxifloxacin will be temporary halted, and a review of available safety data will 
be conducted by the SMC if any of the following occur after dosing: 
1. Death of a subject following dosing regardless of relatedness to moxifloxacin;  
2. One subject with a non-fatal SAE that is considered related to moxifloxacin by the study PI or 
authorized physician (listed in FDA Form 1572); 
3. Three or more subjects with a Grade 3 systemic AE coded in the same organ system that is 
considered related to moxifloxacin by the study PI or authorized clinician.  
Note: VS abnormalities will be considered part of a systemic disorder or an organ specific condition, as described 
in Appendix C: Adverse Events Toxicity Grading Criteria 
, in order to be included among the Study Halting Criteria. 
9.6 
Safety Oversight 
9.6.1 
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. The ISM will review SAEs and other AEs as 
needed and provide an independent assessment to DMID. 
9.6.2 
Safety Monitoring Committee (SMC) 
Safety oversight will be conducted by a SMC, which is an independent group of experts that 
monitors subject safety and advises DMID. SMC members will be separate and independent of 
study personnel participating in the trial and will not have scientific, financial, or other conflicts 
of interest related to the trial. The SMC will consist of members with appropriate expertise to 
contribute to the interpretation of the data from the trial. 
The SMC will meet as follows: 
• Organizational meeting (before study initiation) 
• Ad hoc meeting 
o When study halting criteria are met 
o At the request of DMID to review a potential safety concern identified by either the 
Site PI, ISM, or DMID MM 
• Scheduled meeting 

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o The SMC will review cumulative, unblinded interim safety data when available after 
database lock 
Procedures for SMC reviews/meetings will be defined in the SMC charter. The SMC will review 
safety data and advise on resumption or stopping of a temporary halted trial, and safety 
monitoring in future clinical trials with zoliflodacin.  

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10 
CLINICAL MONITORING 
10.1 
Site Monitoring Plan 
Site monitoring will be conducted to ensure that human subject protections, study and laboratory 
procedures, study intervention administration, and data collection processes are of high quality 
and meet DMID, GCP/ICH, and regulatory guidelines, when appropriate. Site visits may be 
conducted by an authorized representative of DMID or other regulatory agencies to inspect study 
data, subjects’ medical records, and eCRFs in accordance with ICH guidelines, GCP, and the 
respective local and national government regulations and guidelines. 
The investigator will permit authorized representatives of DMID and the respective local and 
national health authorities to inspect facilities and records relevant to the trial, if needed. 
A separate monitoring plan developed by DMID will describe protocol-specific items to be 
monitored. 
Site visits will be made at standard intervals defined by DMID and may be made more frequently 
as directed by DMID. Monitoring visits will include, but not be limited to, review of regulatory 
files, accountability records, subjects’ source documents, eCRFs, ICFs, clinical and laboratory 
reports, and protocol and GCP compliance. Site monitors will have access to the CTU, study 
personnel, and all study documentation according to the DMID-approved site monitoring plan. 
Study monitors will meet with the Site PI 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 
The trial will evaluate the following hypotheses: 
a. Zoliflodacin administered at 2-g or 4-g doses causes a prolongation of the QTcF interval 
that exceeds the criteria in the ICH Guidance E14.16  
(A negative effect is defined in ICH Guidance E14 as “one in which the upper bound of the 
one-sided 95% confidence interval (CI) for the largest time-matched mean effect of the drug 
on the QTcF interval is <10 msec.”) 
b. The trial has the sensitivity to detect an effect on QTcF of regulatory concern, as produced 
by moxifloxacin. 
 
(Assay sensitivity will be demonstrated if, for at least one individual timepoint between 1h 
and 4h after administration of moxifloxacin, the lower bound of the one-sided 95% CI for 
ΔΔQTcF, is >5 msec.) 
11.2 
Sample Size Considerations 
Enrollment of 72 subjects would provide at least 84% power to conclude a negative effect, given 
that up to 16 subjects may withdraw early prior to beginning to replace subjects (at least 56 
subjects evaluable), and assuming a standard deviation of ΔΔQTcF of 7 msec and an underlying 
effect of 5 msec. 
11.3 
Safety Review 
An SMC will be appointed to oversee the safe conduct of the trial. A scheduled SMC meeting 
will be held after all subjects complete the trial to review safety data and advise on safety 
monitoring in future clinical trials of zoliflodacin. If criteria for halting the trial (as listed in 
Section 9.5.1) are met, an ad hoc SMC meeting will be held to review all available safety data and 
to make recommendations about the dosing of all further subjects in the trial. An independent 
unblinded statistician may present data in a closed session of an SMC meeting. 
11.4 
Final Analysis Plan 
The ICH Guidance E9 (Statistical Principles for Clinical Trials) will be followed for all statistical 
content.24 Summaries of frequencies and percentages will be presented for categorical data, and 
summary statistics such as minimum, median, mean, SD, and maximum values will be presented 
for continuous data. Details of safety, QTcF, other ECG interval measurements (QT, PR, QRS, 
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PK/PD data analyses and presentations will be described in the Statistical Analysis Plan (SAP) 
and accompanying Tables, Listings, and Figures (TLF) templates. The SAP for safety and PK 
data will be prepared and finalized by the DCC before final data lock. The SAP for Holter ECG 
data will be prepared by IQVIA Cardiac Safety Services (CSS) and appended to the SAP. 
Analyses included in the clinical study report (CSR) will be performed by the DCC and CSS after 
final data lock. Any change from originally planned statistical analyses will be reported in the 
CSR.   
11.4.1 
Analysis Populations 
All subjects who received at least one dose of study drug (zoliflodacin, placebo, or moxifloxacin) 
will be included in the safety population and analyzed as treated.  
The PK analysis population will consist of all subjects who received zoliflodacin and have at least 
one quantifiable post-dosing drug concentration measured. The PK analysis subset will be based 
on the PK population, which includes all subjects who completed the PK part of the trial without 
any protocol violations that would likely affect the PK results and who have evaluable plasma 
concentration data for zoliflodacin from whom at least a subset of the designated PK parameters 
can be determined. If any subjects are found to be noncompliant with respect to dosing or have 
incomplete data, a decision to include them in the analysis will be made on a case-by-case basis.  
The PD analysis population will include all subjects who provide evaluable Holter ECG data from 
at least one study treatment, without protocol deviations potentially impacting the ECG intervals. 
Exclusions from the PD analysis population due to protocol deviations will be decided before 
unblinding. Subjects must have at least one ∆∆QTcF value defined for inclusion in the PD 
analysis population.   
The PD and PK analysis population will include all subjects who provide both evaluable Holter 
ECG and zoliflodacin PK data for the relevant study treatment. The PK/QTcF population will be 
the subset of subjects included in both the PD analysis population and the PK analysis population. 
Additionally, for the concentration-QTcF analysis, time-matched zoliflodacin concentrations 
corresponding to the ECG assessments will also be necessary. 
11.4.2 
Demographics and Baseline Characteristics 
Subject demographics and baseline characteristics will be summarized. The number of subjects 
who enroll in the trial, and the number and percentage of subjects who complete each assessment, 
will be presented. The percentage of subjects who withdraw from the trial or discontinue the 
study drug, and reasons for withdrawal or discontinuation, will be summarized. 

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11.4.3 
Safety Analysis 
11.4.3.1 
Adverse and Serious Adverse Events 
AEs will be coded using the latest version of the Medical Dictionary for Regulatory Activities® 
(MedDRA). All AEs occurring after study drug dosing will be summarized using frequency 
counts and percentages. The following summaries will be presented for AEs and SAEs:  
• Overall (i.e., regardless of severity or relationship to treatment); 
• By severity grade (mild, moderate, or severe); 
• By relationship to study drug; 
• By MedDRA level hierarchy (system organ class [SOC], higher level group term [HLGT] 
and preferred term [PT]). 
Unless otherwise specified, at each level of subject summarization in reporting the incidence of 
AEs, a subject will be counted once if the subject reported one or more AEs. If more than one 
occurrence of an AE is reported, the AE of the worst severity or the worst-case relationship 
assessment will be summarized.  
11.4.3.2 
Additional Safety Analyses 
Descriptive summary statistics (mean, SD, median, minimum, and maximum) for clinical 
laboratory data, 12-lead standard ECG and Holter ECG parameters, and VS at admission and each 
applicable post-dosing visit, including changes from baseline values, will be calculated. If 
multiple baseline values are obtained, only the most recent value will be analyzed (with the 
exception of Holter ECG, see below). For change-from-screening summaries, subjects with an 
undefined change from screening, due to missing data, will be excluded. Clinical significance of 
abnormalities will be indicated in the listings. 
11.4.4 
PK Analysis  
PK parameters will be estimated for zoliflodacin by noncompartmental methods using version 8.0 
or higher of Phoenix® WinNonlin®. When evaluable, estimated PK parameters will include: 
• AUC(0-last): Area under the concentration time-curve from time zero to the last 
concentration above the lower limit of quantitation 
• AUC(0-∞): Area under the concentration time-curve from time zero to infinity  
• AUC(0-t): Area under the concentration time-curve from time zero to time t 
• Cmax: Maximum observed concentration 
• Tmax: Time of maximum observed concentration 
• Ke: Elimination rate constant  
• t1/2: Terminal elimination half-life 
• CL/F: Apparent oral clearance 
• Vz/F: Apparent volume of distribution 
Other PK parameters may be calculated, as appropriate. 

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The results will be listed by subject, and summarized with descriptive statistics including: n, 
mean, SD, coefficient of variation, median, minimum, maximum, geometric mean, and geometric 
SD.  
Graphical presentations of concentration vs. time profiles will be provided for zoliflodacin and 
will include individual subject and mean concentration profiles. Semi-log concentration profiles 
will be provided for individual subjects. 
11.4.5 
Missing Values and Outliers 
All attempts will be made to collect all data per protocol. No imputation will be performed for 
missing values. Outliers will not be excluded from the primary analyses. Outliers identified 
during the PK analysis will be discussed in the analysis report. 
11.4.6 
Analysis of Continuous ECG Parameters (Pharmacodynamics) 
11.4.6.1 
ECG Analysis and Interpretation 
The ∆∆QTcF, calculated as differences between time-matched mean change-from-baseline QTcF 
values for active treatment relative to placebo, will be evaluated to quantify the QTcF 
prolongation of the active treatments. The median of the three Holter ECG QTcF interval 
measurements at 45, 30, and 15 min before each dosing will be regarded as baseline for the 
respective period. 
QTcF corrections will be calculated using Fridericia’s formula: 
• QTcF=QT/RR0.33 
The primary variable will be the change-from-baseline QTcF (∆QTcF). Other endpoints to be 
evaluated include HR (bpm; calculated from RR interval measurements), PR (msec), QRS (msec), 
RR (msec), and cardiac abnormalities (morphologies, rhythm, and conduction).   
All statistical analyses and reporting of digital ECG data will be conducted on the mean of the 
replicate intervals within a timepoint. Descriptive statistics including two-sided 90% CI will be 
generated for observed, change-from-baseline, and time-matched placebo-corrected change-from-
baseline ECG intervals. 
11.4.6.2 
Primary Analysis: QTcF Effect 
The primary hypothesis to be tested is that following administration of zoliflodacin 2 g and 4 g, 
the upper bound of the one-sided 95% CI of treatment effect on ∆∆QTcF is ≥10 msec for at least 
one of the ECG assessments, against the alternative hypothesis that all mean effects are <10 msec. 
The primary analyses will be conducted to evaluate the effects of zoliflodacin 2 g and 4 g on 
cardiac repolarization as measured by ∆QTcF relative to placebo. Analysis will be performed 
using a linear mixed model with ∆∆QTcF as the dependent variable, with baseline QTcF, 
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effects, and subject nested within sequence as random effects. At each of the 9 post-baseline 
timepoints and for each zoliflodacin dose, a one-sided 95% CI for the treatment effect on 
∆∆QTcF will be estimated using least square means. There will be 18 CIs, one for each 
combination of zoliflodacin dose and timepoint. A negative effect of 2 g or 4 g zoliflodacin will 
be concluded if the maximum of the 18 one-sided 95% CI upper bound is <10 msec. No 
adjustment will be made for multiplicity, since it is not required under intersection-union testing, 
as planned for the trial. No subgroup analyses are planned. 
11.4.6.3 
Secondary Analyses: Non-QTc Intervals 
Non-QTc intervals (PR, QRS, and RR intervals) and HR (estimated from measurement of RR 
intervals) will be tabulated at each timepoint for each treatment group and the mean ± SD, 
median, 95% CI, minimum, and maximum values will be shown. Comparisons among treatment 
groups will be done using linear mixed effects models.  
Categorical analysis of PR, QRS, and RR intervals will also be performed. The following criteria 
will be used to specify noteworthy non-QTc intervals (PR, QRS, and RR intervals) and HR 
(derived from RR intervals) and the number of subjects and number of timepoints will be 
tabulated for each treatment group and may be compared statistically: 
• 
PR change-from-baseline >25% resulting in PR >200 msec  
• 
QRS change-from-baseline >25% resulting in QRS >120 msec 
• 
HR change-from-baseline >25% decrease resulting in HR <50 bpm 
• 
HR change-from-baseline >25% increase resulting in HR >100 bpm 
11.4.6.4 
Assay Sensitivity 
Another secondary hypothesis is that a moderate increase in QTcF with administration of the 
positive control moxifloxacin 400 mg is detectable in this trial.   
The model as described in the primary analyses will be repeated with the moxifloxacin treatment 
for assay sensitivity evaluation. The 1, 2, 3, and 4 h timepoints will be evaluated for assay 
sensitivity. Type I error (familywise error rate) will be controlled at an overall level of 5% by 
using a prespecified multiplicity adjustment. Assay sensitivity will be demonstrated if the lower 
bound of one-sided 95% CI (bounds will vary based on multiplicity adjustment) for treatment 
effect on ∆∆QTcF in at least one timepoint is >5 msec. In addition, two-sided 90% CIs will be 
obtained for the treatment effect on ∆∆QTcF at every timepoint and displayed graphically. 
11.4.6.5 
Categorical Analyses 
QTcF intervals will be categorized and tabulated for each treatment. The following criteria will be 
used to specify noteworthy QTcF intervals:  
• QTcF >450 msec  
• QTcF >480 msec  

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• QTcF >500 msec  
• QTcF increase ≥30 msec from baseline 
• QTcF increase ≥60 msec from baseline 
The number and percent of subjects in each category will be calculated. Comparisons across 
treatments may be conducted if sufficient counts are observed.  
11.4.6.6 
Cardiac Abnormalities: 
The morphological waveform analysis of the ECG data for TQT evaluations at each timepoint 
will be utilized for the summary of new onset cardiac abnormalities for each treatment. Results 
will be summarized in frequency tables with counts and percentages for both number of subjects 
and number of timepoints. 
11.4.7 
Concentration-QTcF Modeling 
Concentration-QTcF modeling will be utilized to characterize the exposure-QTcF relationship for 
zoliflodacin.25,26 Initially, linear or non-linear mixed-effect models will be fit to ∆∆QTcF values 
for only the zoliflodacin arms. More complex models will be employed if simpler models provide 
inadequate model fit. Hysteresis effects will be examined and may be utilized to characterize the 
delayed effects if observed. Estimates of the mean prolongation at the geometric mean 
zoliflodacin Cmax for the zoliflodacin 2-g and 4-g doses will be made from the final exposure 
response model with corresponding two-sided 90% CIs.   
Detailed analysis plans will be documented in the SAP. Results of the concentration-QTcF 
evaluations will be reported in the CSR. 

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12 
SOURCE DOCUMENTS AND ACCESS TO SOURCE 
DATA/DOCUMENTS 
The CTU will maintain appropriate medical and/or research records for the trial, in compliance 
with ICH E6 GCP, Section 4.9, and regulatory and institutional requirements for the protection of 
confidentiality of subjects. As part of participating in a DMID-sponsored, DMID-affiliated, or 
manufacturer-sponsored trial, the CTU will permit authorized representatives of DMID, to 
include Emmes (the DCC), DynPort Vaccine Company, LLC (DVC), and regulatory agencies to 
review (and, when required by applicable law, to copy) clinical records for the purposes of quality 
assurance reviews, audits, and evaluation of the study safety and progress. 
Forms for use as source documents will be derived from eCRFs and will be provided by the DCC 
and CTU. Additional 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. Examples of these original documents and data records include, but are not limited to, 
hospital records, clinical and office charts, laboratory notes, memoranda, 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, ECG print-outs, and subject files and records kept at the pharmacy, 
laboratories, and medico-technical departments involved in the trial. 
 

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13 
QUALITY CONTROL AND QUALITY ASSURANCE 
Following a written DMID-accepted site quality management plan, the CTU 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 the CTU, source 
data/documents, and reports for monitoring and auditing by DMID, and inspection by local and 
regulatory authorities. The PI will ensure all study personnel are appropriately trained and 
applicable documentations are maintained on site.  
The DCC 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 CTU for clarification and resolution. 

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14 
ETHICS/PROTECTION OF HUMAN SUBJECTS 
14.1 
Ethical Standard 
The PI will ensure that the 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 25692 (1997), if applicable. The PI’s institution will hold a current Federal Wide 
Assurance (FWA) issued by the Office of Human Research Protection for federally-funded 
research. 
14.2 
Institutional Review Board 
The CTU will provide for the review and approval of this protocol and associated ICFs by an 
appropriate IRB/EC listed on the FWA. Any amendments to the protocol or consent materials will 
also be approved before they are used, unless change is for the safety of the subject. Only those 
IRB members who are independent of the investigators and DMID will provide an opinion on 
study-related matters. Verification of IRB approval of the protocol and the written ICF will be 
transmitted by the investigator or designee before shipment of the study drug. No deviations from 
or changes to the protocol will be initiated without prior approval of an appropriate amendment 
unless change is for the safety of the subject. 
14.3 
Informed Consent Process 
The written consent document will embody the elements of informed consent as described in the 
Declaration of Helsinki and will adhere to the ICH Harmonized Tripartite Guideline for GCP. 
Informed consent will be obtained before any protocol-specified procedures or interventions are 
carried out, and in accordance with 21 CFR 50.25 and 45 CFR 46. Information will be presented 
both orally and in written form. 
An investigator or designee will describe the protocol to potential subjects face-to-face. The ICF 
may be read to the subjects, but, in any event, the investigator shall give the subjects ample 
opportunity to inquire about details of the trial and ask any questions before signing the ICF. 
Study staff will inform subjects that the trial involves research, and explain the purpose of the 
trial, those aspects of the trial that are experimental, any expected benefits, all possible risks 
(including a statement that the particular treatment or procedure may involve risks to the subject 
or to the embryo or fetus, if the subject is or may become pregnant, that are currently 
unforeseeable), the expected duration of the subject’s participation in the trial, the procedures of 
the research study, including all invasive procedures, and the probability for random assignment 

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to treatment groups. Subjects will be informed that they will be notified in a timely manner if 
information becomes available that may be relevant to their willingness to continue participation 
in the trial. They will also be informed of alternative procedures that may be available, and the 
important potential benefits and risks of these available alternative procedures. Subjects will 
receive an explanation as to whether any compensation and any medical treatments are available 
if injury occurs, and, if so, what they consist of, or where further information may be obtained. 
Subjects will be informed of the anticipated financial expenses, if any, to the subject for 
participating in the trial, as well as any anticipated prorated payments, if any, to the subject for 
participating in the trial. They will be informed of whom to contact (e.g., the investigator) for 
answers to any questions relating to the research project. Information will also include the 
foreseeable circumstances and/or reasons under which the subject’s participation in the trial may 
be terminated. Subjects will be informed that participation is voluntary and that they are free to 
withdraw from the trial for any reason at any time without penalty or loss of benefits to which the 
subject is otherwise entitled. 
Neither the investigator, nor the trial staff, will coerce or unduly influence a subject to participate 
or continue to participate in the trial. The extent of the confidentiality of the subjects’ records will 
be defined, and subjects will be informed that applicable data protection legislation will be 
followed. Subjects will be informed that the monitor(s), auditor(s), IRB, DMID, and regulatory 
authority(ies) will be granted direct access to the subject’s original medical records for 
verification of clinical trial procedures and/or data without violating the subject’s confidentiality, 
to the extent permitted by applicable laws and regulations, and that, by signing a written ICF, the 
subject is authorizing such access. Subjects will be informed that records identifying the subject 
will be kept confidential, and, to the extent permitted by the applicable laws and/or regulations, 
will not be made publicly available and, if the results of the trial are published, the subject’s 
identity will remain confidential. 
ICFs will be in a language fully comprehensible to the prospective subjects. Informed consent 
shall be documented using a written consent form approved by the IRB and signed and dated by 
the subject and the person who conducted the informed consent discussion. The signature 
confirms that the consent is based on information that has been provided and all questions have 
been answered to the prospective participant’s satisfaction. Each subject’s signed ICF will be kept 
on file by the investigator for possible inspection by regulatory authorities and/or DMID and 
regulatory compliance persons. The subject will receive a copy of the signed and dated written 
ICF and any other written information provided to the subjects and will receive copies of any 
signed and dated ICF updates and any amendments to the written information provided to 
subjects. 

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14.4 
Exclusion of Women, Minorities, and Children (Special Populations) 
Children aged <18 years will be excluded from participation because insufficient data are 
available in adults to judge potential risk in children, and as a Phase 1 trial, there is no known 
benefit.   
Neither women nor minorities will be routinely excluded from participation in the trial. Subjects 
will be recruited without regard to gender or race. It is expected that race and gender distributions 
in the trial will approximate the proportion to their numbers within the community.   
Women of childbearing potential will be included but will be repeatedly counseled to use 
effective measures (Section 5.1) to avoid becoming pregnant from the time of screening until 30 
days after the last dose of study drug is received because the effects of the study drug on the 
unborn fetus are not known.    
14.5 
Subject Confidentiality 
Subject confidentiality is held strictly in trust by the participating investigators, their staff, and 
DMID and its agents. This confidentiality is extended to cover testing of biological samples, and 
also clinical information related to participating subjects. 
The study protocol, documentation, data, and all other information generated will be held in strict 
confidence. No information concerning the trial or data will be released to any unauthorized third 
party without prior written approval from DMID. This information and data will not be used by 
the Site PI or other study personnel for any purpose other than conducting the trial. These 
restrictions do not apply to: (1) information which becomes publicly available through no fault of 
the Site PI or other study personnel; (2) information which is necessary to disclose in confidence 
to an IRB solely for the evaluation of the trial; (3) information which is necessary to disclose in 
order to provide appropriate medical care to a subject; or (4) study results which may be 
published as described in Section 16. 
The study monitors or other authorized representatives of DMID may inspect all documents and 
records required to be maintained by the Site Investigator, including, but not limited to, medical 
records (office, clinic, or hospital) and pharmacy records for the subjects in the trial. The CTU 
will permit access to such records. 
14.6 
Certificate of Confidentiality 
To protect privacy, we have received a Certificate of Confidentiality.  With this Certificate, the 
researchers cannot be forced to release information that may identify the research subject, even by 
a court subpoena, in any federal, state, or local civil, criminal, administrative, legislative, or other 
proceedings.  The researchers will use the Certificate to resist any demands for information that 
would identify the subject, except as explained below. 

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The Certificate cannot be used to resist a demand for information from personnel of the United 
States Government that is used for auditing or evaluation of federally funded projects, like this 
study, or for information that must be released in order to meet the requirements of the Federal 
Food and Drug Administration (FDA). 
A Certificate of Confidentiality does not prevent the subject from voluntarily releasing 
information about themselves or their involvement in this research.  If any person or agency 
obtains a written consent to receive research information, then the researchers may not use the 
Certificate to withhold that information.   
The Certificate of Confidentiality does not prevent the researchers from reporting without the 
subject’s consent, information that would identify the subject as a participant in the research 
project regarding matters that must be legally reported including: child and elder abuse, sexual 
abuse, or wanting to harm themselves or others.   
The release of individual private information or specimens for other research will only occur if 
consent was obtained from the individual to whom the information, document, or biospecimen 
pertains, or for the purposes of other research that is in compliance with applicable Federal 
regulations governing the protection of human subjects in research. 
14.7 
Study Discontinuation 
DMID has the right to terminate the trial or the CTU’s participation at any time. Reasons for 
terminating the trial may include, but are not limited to: 
• Incidence or severity of AEs indicates a potential health hazard 
• Data recording is inaccurate or incomplete 
• Investigator does not adhere to the protocol or applicable regulatory guidelines in 
conducting the trial 
If the trial is discontinued, subjects who have signed the ICF and received the study drug will 
continue to be followed for safety for the duration of the trial. No further study treatments will be 
administered to other subjects. 

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15 
DATA HANDLING AND RECORD KEEPING 
The Site PI is responsible to ensure the accuracy, completeness, legibility, and timeliness of 
reported data. All data collection forms will be completed legibly to ensure accurate interpretation 
of data. Black or blue 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. 
Data collection forms will be provided by the DCC and the CTU and will be used by the CTU to 
record and maintain data for each subject enrolled in the trial. Data reported in the eCRF derived 
from source documents will be consistent with the source documents or the discrepancies will be 
explained. 
DMID and/or its designee will provide guidance to investigators and other study personnel on 
making corrections to the data collection forms, source documents, and eCRFs. 
15.1 
Data Management Responsibilities 
All source documents and laboratory reports will be reviewed by the clinical team and data entry 
staff, who will ensure that they are accurate and complete. AEs will be graded, assessed for 
severity and causality, and reviewed by the Site PI or designee. 
Data collection is the responsibility of the clinical trial staff at the CTU under the supervision of 
the Site PI. During the trial, the investigator will maintain complete and accurate documentation 
for the trial. 
Emmes will serve as the DCC for the trial, and will be responsible for data management, quality 
review, analysis, and reporting of the study data. 
15.2 
Data Capture Methods 
Clinical data (including, but not limited to AE/SAEs, ConMeds, MH, and PE) and clinical 
laboratory data will be entered into a 21CFR11-compliant Internet Data Entry System provided 
by the DCC. The data system includes password protection and internal quality checks (e.g., 
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 study 
personnel.  
ECG data captured by 12-lead Holter monitors will be transmitted to the ECG Core Lab. QT, 
QTcF, and other ECG interval measurements and ECG interpretations will be performed at the 
ECG Core Lab using validated protocols. ECG data management is provided by IQVIA Cardiac 
Safety Services. 

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15.3 
Types of Data 
Data for the trial will include clinical safety assessments, laboratory safety assessments, 12-lead 
standard ECGs, Holter ECG interval measurements and waveform assessments, and PK 
parameters. 
15.4 
Timing/Reports 
A final CSR will be prepared after all safety, 12-lead standard ECG, 12-lead Holter ECG, and PK 
data are available. See Section 9.6.2 for additional reporting requirements.  
15.5 
Study Records Retention 
Study files and ICFs will be maintained for at least 2 years after the last approval of a marketing 
application in an ICH region and until there are no pending or contemplated marketing 
applications in an ICH region, or at least 2 years have elapsed since the formal discontinuation of 
clinical development of the study drug. These documents will be retained for a longer period, 
however, if required by local regulations. No records will be destroyed without the written 
consent of DMID, if applicable. It is DMID’s responsibility to inform the investigator when these 
documents no longer need to be retained. 
15.6 
Protocol Deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, GCP, or protocol-
specific MOP requirements. The noncompliance may be either on the part of the subject, the 
investigator, or the CTU staff. Corrective actions for protocol deviations are to be developed by 
the CTU and implemented promptly. 
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 PI/study staff to use continuous vigilance to identify and report 
deviations within 5 working days of identification of the protocol deviation, or within 5 working 
days of the scheduled protocol-required activity. All deviations will be promptly reported to 
DMID via DCC’s AdvantageEDCSM. 
All protocol deviations, as defined above, will be addressed in subject source documents. A 
completed copy of the DMID Protocol Deviation Form will be maintained in the regulatory file, 
as well as in the subject's source document. Protocol deviations will be sent to the local IRB/EC 

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per their guidelines. The Site PI/study staff is responsible for knowing and adhering to their IRB 
requirements. 

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16 
PUBLICATION POLICY 
Following completion of the study, the lead Principal Investigator is expected to publish the 
results of this research in a scientific journal. 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 Office of Extramural Research (OER) Grants and Funding, 
http://grants.nih.gov/grants/oer.htm  
 
As of January 2018, all clinical trials supported by the NIH must be registered on 
ClinicalTrials.gov, no later than 21 days after the enrollment of the first subject. Results of all 
clinical trials supported by the NIH, generally, need to be submitted no later than 12 months 
following the primary completion date. A delay of up to 2 years is available for trials that meet 
certain criteria and have applied for certification of delayed posting. As part of the result posting, 
a copy of this protocol (and its amendments) and a copy of the Statistical Analysis Plan will be 
posted on ClincialTrials.gov. For this trial the responsible party is DMID/NIAID/NIH, which will 
register the trial and post results.   
 
Refer to: 
• Public Law 110-85, Section 801, Clinical Trial Databases 
• 42CFR11 
• NIH NOT-OD-16-149 
 
 

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17 
LITERATURE REFERENCES 
1. Unemo M, Nicholas RA. Emergence of multidrug–resistant, extensively drug resistant and 
untreatable gonorrhea. Future Microbiol 2012; 7(12):1401-22.   
   
2. Barbee LA, Dombrowski JC. Control of Neisseria gonorrhoeae in the era of evolving 
antimicrobial resistance. Infect Dis Clin North Am 2013; 27(4):723-37. 
 
3. World Health Organization, Department of Reproductive Health and Research. Global incidence 
and prevalence of selected curable sexually transmitted infections - 2008. Available at: 
http://www.who.int/reproductivehealth/publications/rtis/stisestimates/en/index.html   
(accessed 2 Jan 2018) 
 
4. Centers for Disease Control and Prevention, (CDC). CDC Fact Sheet: STD Trends in the United 
States: 2011 National Data for Chlamydia, Gonorrhea, and Syphilis, 2013. Available at: 
http://www.cdc.gov/std/stats11/trends-2011.pdf (accessed 2 Jan 2018) 
 
5. Centers for Disease Control and Prevention (CDC). Antibiotic Resistance Threats in the United 
States, 2013. Available at:  
https://www.cdc.gov/drugresistance/threat-report-2013/pdf/ar-threats-2013-508.pdf  
(accessed 2 Jan 2018) 
 
6. Unemo M, Shafer WM. Antibiotic resistance in Neisseria gonorrhoeae: origin, evolution, and 
lessons learned for the future. Ann N Y Acad Sci 2011; 1230:E19-28. 
 
7. World Health Organization. Antimicrobial resistance: global report of surveillance - 2014. 
Available at: 
http://apps.who.int/iris/bitstream/10665/112642/1/9789241564748_eng.pdf 
 (accessed 2 Jan 2018)  
 
8. Investigator’s Brochure. Product Name, Zoliflodacin (Research number 
 
. 
 
9. Basarab GS, Kern GH, McNulty J, et al. Responding to the challenge of untreatable gonorrhea: 
ETX0914, a first-in-class agent with a distinct mechanism-of-action against bacterial Type II 
topoisomerases. Sci Rep 2015; 5:11827. 
 
10. Huband MD, Bradford PA, Otterson LG, et al. In vitro antibacterial activity of AZD0914, a new 
spiropyrimidinetrione DNA gyrase/topoisomerase inhibitor with potent activity against Gram-
positive, fastidious Gram-Negative, and atypical bacteria. Antimicrob Agents Chemother 2015; 
59(1):467-74. 
  
11. Su X-H, Wang B-X, Le W-J, et al. Multidrug-Resistant Neisseria gonorrhoeae Isolates from 
Nanjing, China, Are Sensitive to Killing by a Novel DNA Gyrase Inhibitor, ETX0914 
(AZD0914). Antimicrob Agents Chemother 2016; 60(1):621-623. 
 

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12. A Study to Assess the Safety, Tolerability and Pharmacokinetics of AZD0914 (Phase 1 study). 
ClinicalTrials.gov Identifier: NCT01929629. 
https://clinicaltrials.gov/ct2/show/NCT01929629?term=AZD0914&rank=3  
(accessed 27 July 2017). 
 
13. Randomized, Open-label Phase 2 Study of Oral AZD0914 in the Treatment of Gonorrhea. 
ClinicalTrials.gov Identifier: NCT02257918. 
https://clinicaltrials.gov/ct2/show/NCT02257918?term=AZD0914&rank=1  
(accessed 27 July 2017) 
 
14. Coumel P, Maison-Blanche P. Heart rate variability and QT interval: their relationships with the 
cardiac frequency. In Aubert AE, Ector H, Stroobandt R (eds.), 1994. Cardiac Pacing and 
Electrophysiology. Dordrecht: Kluwer Academic Publishers, pp. 63-71. 
 
15. Lecocq B, Lecocq V, Jaillon P. Physiologic relation between cardiac cycle and QT duration in 
healthy volunteers. Am J Cardiol 1989; 64(8):481-6. 
 
16. Guidance for Industry. E14:  Clinical Evaluation of QT/QTc Interval Prolongation and 
Proarrhythmic Potential for Non-Antiarrhythmic Drugs. FDA, October 2005. 
https://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/U
CM073153.pdf (accessed 27 July 2017) 
 
17. Woosley RL, Heise CW and Romero KA. www.Crediblemds.org, QTDrug Lists, Accessions Date 
19 December 2017, AZCERT, Inc., 1822 Innovation Park Drive, Oro Valley, AZ 85755. 
 
18. Moxifloxacin hydrochloride [package insert]. Available at:  
https://dailymed.nlm.nih.gov/dailymed/fda/fdaDrugXsl.cfm?setid=71b02da1-3175-1db8-
192a-c0a8a6cd98a5&type=display   
(Accessed 25 May 2018) 
 
19. Florian et al. Population Pharmacokinetic and Concentration-QTc Models for 
Moxifloxacin: Pooled Analysis of 20 TQT studies. J Clin Pharmacol 2011; 51:1152. 
 
20. Bloomfield DM, et al (2008).  The Effect of Moxifloxacin on QTc and Implications for the Design 
of Thorough QT Studies. Clin Pharmacol Therapeut 2008; 84:475-80. 
 
21. Darpo B. The thorough QT/QTc study 4 years after the implementation of the ICH E14 guidance. 
Br J Pharmacol. 2010; 159(1):49-57. 
 
22. Bolon MK. The newer fluoroquinolones. Infect Dis Clin North Am 2009; 23(4):1027-51. 
 
23. U.S. Medical Eligibility Criteria for Contraceptive Use, 2016. CDC MMWR 2016; 65(3):1-108. 
 
24. Guidance for Industry. E9 Statistical Principles for Clinical Trials. FDA, September 1998. 
https://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm0
73137.pdf (accessed 27 July 2017) 
 
25.  Garnett, CE, et al. Concentration-QT Relationships Play a Key Role in the Evaluation of 
Proarrhythmic Risk during Regulatory Review. J Clin Pharmacol 2008; 48:13-18. 

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26. Salvy, V. et al. Update on the Evaluation of New Drugs for the Effect of Cardiac Repolarization in 
Humans in Early Drug Development. British J Pharmacol 2010; 159:34-48. 
 

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18 
APPENDICES

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Appendix A: Schedule of Study Procedures and Evaluations 
Table 3: Schedule of Events 
Study Visit 
Screening1 
Inpatient Periods 1-4 
Outpatient 
Periods 1-4 
Final Visit 
(after Dose 4) 
Unscheduled 
Early 
Termination 
Check-
in 
Dosing 
Follow-up 
Study Day / Assessments 
-21 to -2 
-1 
1 
2 
3 - 7 
8 (± 2) 
 
 
Informed consent 
X 
 
 
 
 
 
 
 
Inclusion/Exclusion criteria2 
X 
X 
X 
 
 
 
 
 
Demographics 
X 
 
 
 
 
 
 
 
Medical history 
X 
 
 
 
 
 
 
 
Medical history update 
 
X 
 
 
 
 
 
 
Prior and ConMeds 3 
X 
X 
X 
X 
X 
X 
X 
X 
Complete PE4 
X 
 
 
 
 
X 
 
X 
Abbreviated PE5 
 
X 
 
 
 
 
 
 
Symptom-directed PE6 
 
 
X 
X 
 
 
X 
 
Height, weight, BMI7 
X 
 
 
 
 
 
 
 
Weight 
 
X 
 
 
 
X 
 
X 
Vital signs8 
X 
X 
X 
X 
 
X 
X 
X 
Clinical labs (HEM, CHEM, 
UA)9 
X 
X 
 
X 
 
X 
X 
X 
Viral serology10 
X 
 
 
 
 
 
 
 
Serum pregnancy test11 
X 
X 
 
 
 
X 
 
X 
Serum FSH level11 
X 
 
 
 
 
 
 
 
Urine toxicology 
X 
X 
 
 
 
 
 
 
Alcohol urine test 
X 
X 
 
 
 
 
 
 
12-lead standard ECG12 
X 
X 
X 
X 
 
X 
X 
X 
Continuous Holter ECG13  
 
 
X 
X 
 
 
 
 
Randomization14 
 
 
X 
 
 
 
 
 
Study drug dosing 
 
 
X 
 
 
 
 
 

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Study Visit 
Screening1 
Inpatient Periods 1-4 
Outpatient 
Periods 1-4 
Final Visit 
(after Dose 4) 
Unscheduled 
Early 
Termination 
Check-
in 
Dosing 
Follow-up 
Study Day / Assessments 
-21 to -2 
-1 
1 
2 
3 - 7 
8 (± 2) 
 
 
PK samples15 
 
 
X 
X 
 
 
 
X 
Counsel on use of 
appropriate contraception 
and avoidance of pregnancy16 
X 
 
 
X 
 
X 
X 
X 
Counsel to avoid use of 
prohibited medications, 
alcohol, marijuana and illicit 
drugs 
X 
 
 
X 
 
 
 
 
Counsel to avoid rigorous 
physical activity 
X 
X 
 
X 
 
 
 
 
AE and SAE review17 
 
 
X 
X 
X 
X 
X 
X 
Admit to CTU 
 
X 
 
 
 
 
 
 
Discharge from CTU 
 
 
 
X 
 
 
 
 
Discharge from trial 
 
 
 
 
 
X 
 
 
 
1. 
Screening is completed within 21 days before study drug dosing and may require more than one visit. 
2. 
Inclusion/Exclusion criteria at Screening and on Day -1 and Day 1 before dosing in Dosing Period 1 to determine eligibility for enrollment and 
randomization. I/E criteria will be confirmed before each subsequent dosing period.   
3. 
Prior Medications include prescription during the period 30 days before first dosing and non-prescription drugs, herbs, vitamins, and nutritional 
supplements taken during the period 15 days before first dosing. ConMeds include those taken after dosing. (See Section 6.6) for prohibited medications 
and exclusions). 
4. 
Complete PE (except genital, breast, and rectal exams): at Screening, and Final Visit, or ET. 
5. 
Abbreviated PE: on Day -1 (not performed if the complete PE was performed ≤7 days from this visit) in each inpatient period. 
6. 
Symptom-directed PE: on Day 1 (pre-dose and post-dose) and Day 2 in each dosing period to evaluate new symptoms before dosing and AEs after 
dosing. 
7. 
BMI is calculated as wt (kg) / ht (m2). 
8. 
VS (BP, HR, RR, T): at Screening; on Days -1, 1, and 2 in each dosing period; and at Final Visit after Dose 4, or ET. On Day 1 of each dosing period, VS at 
baseline (within 1 h before dosing) and at 1 h (±10 min), 2 h (±10 min), and 4 h (±10 min) after dosing. 
9. 
Clinical laboratory testing with a minimum 4-h fast: HEM (Hgb, Hct, RBC, WBC with differential absolute count, platelet count); CHEM (creatinine with 
estimation of GFR, BUN, glucose, total bilirubin, direct bilirubin, AST, ALT, AP, total protein, albumin, electrolytes (sodium, potassium, chloride, 

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bicarbonate (CO2), magnesium); and dipstick UA (blood, protein, glucose): at Screening, Day -1, and Day 2, or ET. Clinical labs may not be repeated before 
Dose 1 if screening labs were completed ≤7 days before dosing. Clinical labs may be obtained at an unscheduled visit as indicated for work up of an AE. 
10. Viral serology (HIV1/2 test, HBsAg, HCV antibody): at Screening. 
11. Serum pregnancy test in all women at Screening and within 24 h before each dosing period, and at the Final Visit, or ET. FSH only at Screening in post-
menopausal women. 
12. 12-lead standard ECG with 10-sec rhythm strip: at Screening, Day -1, Day 1 [within 1 h before dosing, and at 1 h (±10 min), 2 h (±10 min), and 4 h (±10 min) 
after dosing], Day 2 (24 h ±2 h) or ET. At an Unscheduled Visit, a 12-lead standard ECG will be done only if needed for the evaluation of an AE. 
13. Continuous Holter ECG recorded from 1 h before dosing to 24 h after dosing in each dosing period. ECGs are extracted (three replicates per timepoint) before 
dosing (at 45, 30, and 15 min before dosing) and at 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 12 h, and 24 h after dosing in each dosing period.  
14. Eligible subjects are randomized to a treatment sequence on Day 1 before receiving Dose 1. Randomization confirmed at each dosing period.  
15. Blood (plasma) PK samples: within 30 min before dosing, and at 0.5 h (±5 min), 1 h (±5 min), 2 h (±5 min), 3 h (±10 min), 4 h (±10 min), 6 h (±10 min), 8 h 
(±15 min), 12 h (±15 min), and 24 h (±2 h) after dosing in each dosing period, or ET if it occurs within 24 h of dosing. 
16. Female subjects to use appropriate contraception and avoid pregnancy for 30 days after Final Visit or ET. Male subjects to use appropriate contraception and 
avoid donating sperm for 90 days after Final Visit or ET. 
17. Collect all AEs from the time of dosing in Dosing Period 1 through Final Visit after Dose 4. Follow up AEs and SAEs to resolution or stabilization in 
the clinical judgment of the study investigator.  

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Appendix B: Acceptable Ranges of Screening Laboratory and Vital 
Sign Measurements  
Table 4: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements – 
Hematology and Chemistry 
HEMATOLOGY AND CHEMISTRY  
 
Lab test name 
Reference 
range 
Acceptable 
lower limit 
Acceptable 
upper limit 
Lab unit 
Hemoglobin (Hgb), male 
 13.2 - 17.1 
13.0 
18.5 
g/dL 
Hemoglobin (Hgb), female 
11.7 – 15.5 
11.3 
16.5 
g/dL 
Hematocrit (Hct), male 
 38.5 – 50.0 
38.5 
54 
% 
Hematocrit (Hct), female 
 35 - 45 
35 
49 
% 
White blood cell count (WBC) 
 3,800 – 10,800  
3,500 
11,500 
cells/µL 
Neutrophil count 
 1,500 – 7,800 
1,200 
8,500 
cells/µL 
Lymphocyte count 
 850 – 3,900 
800 
5,000 
cells/µL 
Monocyte count 
200 - 950 
0 
1,100 
cells/µL 
Eosinophil count 
150 - 500 
0 
600 
cells/µL 
Basophil count 
0-200 
0 
300 
cells/µL 
Platelet count 
 140 - 400 
125 
475 
103/µL 
Sodium 
 135 - 146 
132 
148 
mmol/L 
Potassium 
3.5 - 5.3  
3.3 
5.5 
mmol/L 
CO2 (Bicarbonate) 
 20 - 32 
19 
33 
mmol/L 
Chloride 
98 -110  
94 
115 
mmol/L 
Calcium  
 8.6 - 10.3 
8.0 
11.0 
mg/dL 
Magnesium 
 1.5 - 2.5 
1.1 
2.8 
mg/dL 
Blood urea nitrogen (BUN) 
7 – 25  
0 
28 
mg/dL 
Glucose, fasting  
 65 - 99 
50 
115 
mg/dL 
Serum creatinine, male  
0.6 – 1.35 
0.5 
1.45 
mg/dL 
Serum creatinine, female 
0.5 – 1.10 
0.4 
1.2 
mg/dL 
Direct bilirubin 
0.0-0.2  
0 
0.4 
mg/dL 
Total bilirubin  
0.2 – 1.2  
0 
1.4 
mg/dL 
Total protein 
6.1 - 8.1 
5.8 
8.6 
g/dL 
Albumin 
 3.6 – 5.1 
3.2 
6.0 
g/dL 
Aspartate transferase (AST), male 
 10 - 40 
0 
45 
U/L 
Aspartate transferase (AST), female 
10 - 30 
0 
35 
U/L 
Alanine transferase (ALT), male 
9 - 46 
0 
52 
U/L 
Alanine transferase (ALT), female 
5 - 32 
0 
38 
U/L 
Alkaline phosphatase, males 
 40 - 115 
0 
150 
U/L 
Alkaline phosphatase ,females 
33 – 115 
0 
150 
U/L 
Hepatitis B surface antigen (HBsAg) 
non-reactive 
non-reactive 
non-reactive 
 n/a 
Hepatitis C antibody (HCVAb) 
non-reactive 
non-reactive 
non-reactive 
 n/as 
HIV-1/ HIV-2 antibody 
non-reactive 
non-reactive 
non-reactive 
 n/a 

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Serum β-HCG (females only) 
negative 
negative 
negative 
 n/a 
FSH (post-menopausal females 
only) 
23 – 116.3 
23 
116.3 
mIU/mL 
Footnotes: Exceptions to screening laboratory tests’ normal reference ranges are: 
a. 
Racially-based low total WBC or neutrophil counts up to toxicity Grade 1 in Appendix C Toxicity Tables are 
allowed, but toxicity Grades 2 or 3 are exclusionary. 
b. 
Abnormalities in mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular 
hemoglobin concentration (MCHC), red cell distribution width (RDW), mean platelet volume (MPV), and 
nucleated red blood cell count (NRBC CT), which are included in a CBC with differential, will not be 
exclusionary. These results will be included in the database for comparison with results obtained as a part of 
safety laboratory tests.  
c. 
Abnormalities in serum creatinine, BUN, total bilirubin, AST, ALT, and chloride below the lower limit of 
normal (LLN) are allowable. 
d. 
Elevated bilirubin due to documented Gilbert syndrome that is Grade 1 is allowable, but Grade 2 or higher is 
exclusionary. [To document a subject has Gilbert syndrome, a diagnosis from the medical record must will 
be provided or the PI may make a ‘presumptive’ diagnosis of Gilbert syndrome in subjects with 
unconjugated hyperbilirubinemia on repeated testing (at least two samples separated in time) who have 
otherwise normal serum ALT, AST, and AP concentrations, and a normal CBC].  
e. 
Elevated serum glucose, sodium, potassium, bicarbonate (CO2), total protein, and AP values that reach 
toxicity Grade 1 values are allowable. 
f. 
Serum chloride and albumin values above the ULN are allowable. 
g. 
Other laboratory values that are outside the range of eligibility but are thought to be due to an acute condition 
or due to collection or laboratory error may be repeated once.  
 
 
 
 
 
 

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URINALYSIS a 
Table 5: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements – 
Urinalysis 
Lab test name 
Reference 
range limit 
Acceptable 
lower limit 
Acceptable 
upper limit 
Lab unit 
Urine dipstick 
Protein 
negative 
negative 
1+ b 
 
Blood 
negative 
negative 
trace b 
 
Glucose 
negative 
negative 
trace b 
 
Urine Microscopy 
WBCs in urine 
(sediment) 
0-5 
0 
6 
/field 
RBCs in urine 
(sediment) 
0-2 
0 
6 
/field 
Bacteria 
none 
few 
b 
/field 
Hyaline casts 
none 
none 
trace b 
/field 
Crystals 
none 
0 
b 
 
Footnotes:   
a (1) If UA by dipstick is abnormal, a complete UA with microscopic evaluation will be performed and the results will 
supersede the results of the dipstick.  
(2) Menstruating females failing inclusion criteria due to a positive blood on a urine test (dipstick or microscopic UA) may      
be retested following cessation of menses. Do not exclude female subjects with ≤ 6 RBC/HPF. 
b Per investigator judgment, based on medical history. 
 
 
 

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TOXICOLOGY  
 
Urine must be negative for all tested substances [amphetamines, cocaine (and metabolite), 
barbiturates, benzodiazepines, opiates, tetrahydrocannabinol, methamphetamines, 
methadone, TCA, MDMA, and phencyclidine]. Urine creatinine will be measured as part of 
the profile to assess quality of collected sample.  
 
VITAL SIGNS 
Table 6: Acceptable Ranges of Screening Laboratory and Vital Sign Measurements – 
Vital Signs 
 
Measurement 
Lower limit 
Upper limit 
Lab unit 
Systolic BP 
90 
150 
mmHg 
Diastolic BP 
40 
90 
mmHg 
Pulse rate 
45 
100 
bpm 
Respiratory rate 
8 
22 
breaths/min 
Oral temperature 
95.9 
(35.5) 
<100.4 
(<38.0) 
°F 
(°C) 
 
 
 

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Appendix C: Adverse Events Toxicity Grading Criteria 
ABBREVIATIONS: Abbreviations utilized in the Table: 
LLN = Lower Limit of Normal 
Req = Required 
IV = Intravenous 
Dec = Decreased 
ESTIMATING SEVERITY GRADE 
For abnormalities NOT found elsewhere in the Toxicity Tables, use the scale below to 
estimate grade of severity: 
GRADE 1 
Mild 
Events require minimal or no treatment; do not interfere with the 
subject’s daily activities.  
GRADE 2 
Moderate 
Events result in a low level of inconvenience or concern with 
therapeutic measures; may cause some interference with 
functioning.  
GRADE 3 
Severe 
Events interrupt a subject’s usual daily activity and may require 
systemic drug therapy or other treatment; are usually 
incapacitating.  
SERIOUS OR LIFE-THREATENING AEs 
Clinical events considered to be serious or life-threatening include, but are not limited to:  
seizures, coma, tetany, diabetic ketoacidosis, disseminated intravascular coagulation, diffuse 
petechiae, paralysis, acute psychosis, and severe depression. 
COMMENTS REGARDING THE USE OF THESE TABLES 
• Standardized and commonly used toxicity tables (Division of AIDS, National Cancer 
Institute [NCI] Common Toxicity Criteria [CTC], and World Health Organization 
[WHO]) have been adapted for use by the Division of Microbiology and Infectious 
Diseases (DMID) and modified to better meet the needs of subjects in DMID trials. 
• For parameters not included in the following Toxicity Tables, the CTU will refer to the 
“Guide for Estimating Severity Grade” located above. 
• Criteria are generally grouped by body system. 
Some protocols may have additional protocol specific grading criteria, which will supersede 
the use of these tables for specified criteria. 
 
 

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Table 7: Toxicity Grading Tables – Clinical AEs 
Note: Isolated/individual abnormalities of VS would not be considered toward halting criteria. Abnormalities of VS will be 
described as “increased X” or “decreased X” (X = HR, BP, RR, temperature) if asymptomatic, transient and not associated with 
a systemic or organ-specific disorder and coded by MedDRA within the System Organ Class (SOC) “Investigations.” These 
abnormalities will be graded per criteria in Appendix C, but not considered in determining whether study stopping criteria have 
been met. On the other hand, abnormalities of VS that are either secondary to systemic or organ-specific clinical syndrome or 
primary disorders will be coded in the appropriate SOC (e.g., "cardiac disorders", respiratory disorders", "immunological 
disorders", etc.). These abnormalities will be considered in determining whether stopping criteria have been met. 
 
Toxicity Grading Tables 
Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Comments 
VITAL SIGNS 
Fever - °C 
 
Fever - °F 
38.0-38.4 
 
100.4-101.1 
38.5-38.9 
 
101.2-102.0 
>38.9 
 
>102.0 
No recent hot or cold 
beverages or smoking.  
A protocol should select 
either 0C or 0F for 
inclusion 
Tachycardia - bpm 
101-115 
116-130 
>130 or ventricular 
dysrhythmias 
 
Assume awake and in 
supine position for 5 
min at rest; for AE, 
measurements at least 3 
times with 2 concordant 
results (Section 8.1.6) 
Bradycardia – bpm 
• 
Baseline >60 
OR, if 
• 
Baseline <60 
 
• 50-54  
OR, if Baseline <60,  
• 45-50  
 
• 45-49  
OR,  if baseline <60,  
• 
40-44  
 
• <45  
OR,  if baseline <60 
• <40  
As above 
Hypertension (systolic) 
- mmHg 
141-150 
151-160 
>160 
Assume awake, and in 
supine position for 5 
min at rest; for AE, 
measurements on same 
arm at least 3 times 
with 2 concordant 
results (Section 8.1.6) 
Hypertension 
(diastolic) - mmHg 
91-95 
96-100 
>100 
As above 
 Hypotension (systolic) 
- mmHg  
85-89 
80-84 
<80 
As above 
Tachypnea – breaths 
per min 
23-25 
26-30 
>30 
Assume awake and in 
supine position for 5 
min at rest; for AE, 
measurements at least 3 
times with 2 concordant 
results (Section 8.1.6) 

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Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Comments 
CARDIOVASCULAR 
Arrhythmia 
Asymptomatic or 
transient signs; no 
medical intervention 
required 
Recurrent and/or 
persistent signs; 
symptomatic 
medical 
intervention 
required 
Hemorrhage 
Estimated blood loss 
<100 mL 
Estimated blood loss 
>100 mL; no 
transfusion required 
Blood transfusion 
required 
RESPIRATORY 
Cough  
Transient cough; no 
treatment required 
 
Persistent cough; 
treatment required 
Interferes with 
daily activities 
Bronchospasm, Acute 
Transient 
bronchospasm; no 
treatment required; 
FEV1 71-80% of 
predicted peak flow 
Requires treatment; 
normalizes with 
bronchodilator; FEV1 
60-70% of predicted 
peak flow 
  
No normalization 
with 
bronchodilator; 
FEV1 <60% of 
predicted peak flow 
 
Dyspnea   
Does not interfere 
with usual and social 
activities 
Interferes with usual 
and social activities; no 
treatment 
Prevents usual and 
social activities OR 
requires treatment 
GASTROINTESTINAL   
Nausea  
No interference with 
normal activity 
Some interference with 
normal activity 
Prevents daily 
activities 
 
Vomiting  
No interference with 
activity OR 1-2 
episodes in a 24-h 
period 
Some interference with 
activity OR >2 
episodes in a 24-h 
period 
Prevents daily 
activity OR 
requires medical 
intervention 
 
Diarrhea  
2-3 loose or watery 
stools in a 24-h period 
4-5 loose OR watery 
stools in a 24-h period 
6 or more loose or 
watery stools in a 
24-h period OR 
requires IV 
hydration OR 
requires medical 
intervention 

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Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Comments 
Oral Discomfort / 
Dysphagia 
Mild discomfort; no 
difficulty swallowing 
Some limits on eating 
/drinking 
Eating/talking very 
limited; unable to 
swallow solid 
foods 
 
LOCAL IV CATHETER REACTION 
IV site reaction 
Not applicable 
Erythema with 
associated symptoms 
(e.g., edema, pain, 
induration, phlebitis) 
Ulceration or 
necrosis; severe 
tissue damage; 
operative 
intervention 
indicated 
 
SYSTEMIC REACTIONS 
Anaphylaxis ** 
 
 
-- 
-- 
Symptomatic 
bronchospasm, with 
or without urticaria; 
parenteral 
intervention 
indicated; allergy-
related edema or 
angioedema; 
hypotension 
 
**Definition: A disorder characterized by an acute inflammatory reaction resulting from the release of histamine and 
histamine-like substances from mast cells, causing a hypersensitivity immune response. Clinically, it presents with breathing 
difficulty, dizziness, hypotension, cyanosis, and loss of consciousness, and may lead to death. 
Allergic Reaction  
Pruritus without rash 
Localized urticaria OR 
requires oral therapy 
Generalized 
urticaria OR 
angioedema OR 
anaphylaxis OR 
requires 
epinephrine 
 
Hypersensitivity 
(including drug fever) 
Transient flushing or 
rash; temperature 
38.0-38.4°C (100.4-
101.1°F) 
Rash; flushing; 
urticaria; dyspnea; 
temperature 38.5 - 
38.9°C (101.2 – 
102.0°F) 
Symptomatic 
bronchospasm 
with or without 
urticaria; 
parenteral 
medication 
indicated; 
allergy-related 
edema or 
angioedema; 
hypotension; 
temperature 
>38.9°C 
(>102.0°F)  
 

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Clinical AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Comments 
Headache  
No interference with 
activity 
Repeated use of non-
narcotic pain reliever 
for more than 24 h OR 
some interference with 
activity 
Significant; any use 
of narcotic pain 
reliever OR 
prevents daily 
activity OR 
requires triptans 
Fatigue  
No interference with 
activity 
Some interference with 
activity 
Significant; 
prevents daily 
activity 
Myalgia 
No interference with 
activity 
Some interference with 
activity 
Significant; 
prevents daily 
activity 
SKIN 
Mucocutaneous 
Erythema, pruritus 
Diffuse, maculo-
papular rash, dry 
desquamation 
Vesiculation OR 
moist 
desquamation OR 
ulceration 
 
Pruritus 
No or minimal 
interference with usual 
social and functional 
activities  
Greater than minimal 
interference with usual 
social and functional 
activities  
Inability to perform 
usual social and 
functional daily 
activities 
 
ALL OTHER CONDITIONS 
Illness or clinical AE 
(as defined according 
to applicable 
regulations) 
Require minimal or no 
treatment; does not 
interfere with the 
subject’s daily 
activities. 
 
Result in a low level of 
inconvenience or 
concern with 
therapeutic measures; 
may cause some 
interference with 
functioning. 
 
Interrupt a 
subject’s usual 
daily activity and 
may require 
systemic drug 
therapy or other 
treatment; are 
usually 
incapacitating 
 
 
 
 
 
 
 
 
 

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Table 8: Toxicity Grading Tables – Laboratory AEs 
Laboratory AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Blood, serum, or plasma * 
Sodium decrease – mmol/L 
130 - 131 
125 –129 
<125 
Sodium increase – mmol/L 
149 - 150 
 151 – 153 
>153 
Potassium increase – mmol/L 
5.6  – 6.0 
6.1 – 6.4 
≥6.5 
Potassium decrease – mmol/L 
3.0 – 3.2  
2.5 – 2.9 
<2.5 
Bicarbonate (CO2) increase – mmol/L 
34 - 35 
36 - 37 
>37 
Bicarbonate (CO2) decrease – mmol/L 
17 - 18 
14 - 16 
<14 
Glucose decrease, fasting – mg/dL 
47 - 49 
45 –46 
<45 
Glucose increase, fasting – mg/dL 
116 – 125 
126 – 249 
≥250 
Glucose increase, non-fasting – mg/dL 
116 – 160 
161 – 249 
≥250 
Blood urea nitrogen – mg/dL 
25 - 27 
28 – 32 
>32 
Creatinine increase, male – mg/dL 
1.46 – 1.7 
1.71 – 2.3 
>2.3 
Creatinine increase, female – mg/dL 
1.21 – 1.5 
1.51 – 2.0 
>2.0 
Calcium decrease – mg/dL 
7.8 – 8.0 
7.0 – 7.7 
<7.0 
Calcium increase – mg/dL 
11.0 – 11.4 
11.5 – 12.4 
≥12.5 
Total protein decrease – g/dL 
5.1 – 5.7 
4.6 – 5.0 
<4.6 
Albumin decrease – g/dL 
2.8 – 3.4 
2.5 – 2.7 
<2.5 
AST increase, male – U/L 
46 – 115 
116 – 220 
>220 
AST increase, female – U/L 
36 - 90 
91 - 175 
>175 
ALT increase, male – U/L 
53 – 115 
116 - 230 
>230 
ALT increase, female – U/L 
39 – 95 
96 – 190 
>190 
Alkaline phosphatase increase – U/L 
151 – 240 
241 – 360 
>360 
Total bilirubin (serum) increase – mg/dL 
(with other LFTs in the normal range) 
1.5 – 2.0 
2.1 – 2.5 
>2.5 
Total bilirubin (serum) increase – mg/dL 
(accompanied by a >3 x ULN increase in ALT or 
AST)** 
1.5 – 1.7 
1.8 – 2.1 
>2.1 
Hemoglobin decrease, female – g/dL 
10.8 – 11.2 
9.4 – 10.7 
<9.4 
Hemoglobin decrease, male – g/dL 
11.8 – 12.9 
10.0 – 11.7 
<10.0 
WBC increase – cells/µL 
11,000 – 15,000 
15,001 – 20,000 
>20,000 
WBC decrease – cells/µL 
2,500 – 3,500 
1,500 – 2,499 
<1,500 
Neutrophils decrease – cells/µL 
1,200 – 1,499 
1,000 – 1,199 
<1,000 
Lymphocytes decrease – cells/µL 
750 – 849 
500 – 749 
<500 

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Laboratory AEs 
Mild 
(Grade 1) 
Moderate 
(Grade 2) 
Severe 
(Grade 3) 
Monocytes increase – cells/µL 
1,101 – 2,000 
2,001 – 3000 
>3,000 
Eosinophils increase – cells/µL 
500 – 750 
751 – 1,500 
>1,500 
Basophils increase – cells/µL 
201 – 500 
501 – 800 
>800 
Platelets decrease – cells/µL 
120,000 – 124,000 
100,000 – 119,000 
<100,000 
Urine* 
Protein 
1+ 
2+ 
>2+ 
Glucose 
1+ 
2+ 
>2+ 
Blood (dipstick) 
1+ 
2+ 
>2+ 
Blood (microscopic) – RBCs per HPF 
6-10 
11-50 
>50 and/or gross 
blood 
WBC (microscopic) – WBC per HPF 
6-10 
11-50 
>50 
Bacteria (microscopic) 
few 
moderate 
many 
* Institutional normal reference ranges and allowable ranges at screening are provided in Appendix B. 
**ALT, males: >216 U/mL; ALT, females: >132 U/mL; AST, male or female: >126 U/mL) 
Note 1: If a subject was accepted into the trial with a laboratory value of an analyte that overlaps with values used 
for grading Grade 1 laboratory abnormalities, an AE will be reported if the on-study value of the same analyte is 
different (worse) from the baseline. 
Note 2: Safety laboratory results that are abnormal according to the local laboratory reference range, but not 
considered a Grade 1 abnormality, will be evaluated by the CTU clinician and reported as Grade 1 abnormalities if 
clinically significant. If not clinically significant, these will not be considered laboratory AEs and will thus not be 
graded but will be recorded in the source document and followed-up clinically at the discretion of the CTU 
clinician.  
Note 3: Other laboratory parameters performed and reported as parted of the CBC, metabolic panel, and UA will 
be evaluated by the study clinician, recorded in the source document, reported as laboratory AEs if clinically 
significant, and graded according to the criteria in Section 9.2.1. 
Note 4: If the dipstick UA is abnormal, a microscopic UA will be performed, and the results will supersede the 
results of the dipstick UA. 
Note 5: Menstruating females with a positive dipstick UA or microscopic UA may be retested following cessation of 
menses.  
 

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Table 9: Toxicity Grading Tables – ECG 
 
ECG interval 
abnormality  
Grade 1 
Grade 2 
Grade 3 
QTcF interval 
prolonged (msec) 
Asymptomatic, QTcF 
450-479 msec  
Asymptomatic, QTcF 480-500 msec 
OR increase in interval 30-59 msec 
above baseline 
Asymptomatic, QTcF >500 msec 
OR increase in interval ≥60 msec 
above baseline 
PR interval 
prolonged (msec) 
210-250 msec 
>250 msec 
Type II 2nd degree AV block OR 
ventricular pause >3.0 sec 
 
Note: Events will be coded as SAEs if there are life-threatening associated symptoms or signs (arrhythmia, CHF, 
hypotension, syncope, TdP, etc.) 
Note: If a male subject was accepted into the trial with a QTcF value that overlaps with values used for grading 
Grade 1 QTcF prolongation, an AE will be reported if the QTcF value is higher than the baseline value 
 

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Appendix D: Blood Volume Withdrawn During the Trial 
Table 10: Laboratory Samples and Estimated Total Blood Volume (mL)  
Study Periods 
 
Inpatient Periods 1 to 4 
Outpatient Periods 
1 - 4 
Dosing    Period 4 
ET 
 
Study Visit 
Screen 
Check-in 
 Dosing 
Follow-up 
Wash-out 
 
Final 
 
+Study Daya 
-21 to -2 
-1 
1 
2 
3-7 
 
8 (±2) 
 
+ 
 
 
 
 
 
 
 
HEMATOLOGY1 
4 
4 
 
4 
 
4 
4 
CHEMISTRY and serum β-HCG and FSH1 
5 
5 
 
5 
 
5 
5 
Viral Serology (HIV, HBsAg, HCV)1 
8.5 
 
 
 
 
 
 
PK3 
 
 
54 
6 
 
 
6 
Total volume/visit 
17.5 
9 
54 
15 
0 
9 
15 
Dosing periods multiplier 
1 
4 
4 
4 
 
1 
1 
Total volume (all dosing periods) 
17.5 
36 
216 
60 
0 
9 
15 
Cumulative total volume 
17.5 
53.5 
269.5 
329.5 
329.5 
338.5 
 
a Study Days shown correspond to days in each treatment period. For a view of the cumulative numbering of study days, please refer to Section 7. 
  
1 Clinical blood tests (HEM, CHEM) are drawn at Screening Visit, on Day -1, and on Day 2 in each treatment period, or ET.  Serum pregnancy test in all 
women at Screening and within 24 h before each dosing period, and at the Final Visit, or ET. FSH only at Screening in post-menopausal women. 
2 Viral serology tests are drawn at Screening.  

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3 PK serum samples are drawn on Day 1: 30 min before dosing and at 0.5, 1, 2, 3, 4, 6, 8, 12, and 24 h after dosing in each dosing period, or ET if occurs within 
24 h of dosing. 

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Appendix E: A Partial List of Drugs Causing QTc Prolongation and 
Torsade de Pointes 
Table 11: A Partial List of Drugs Causing QTc Prolongation and Torsade de Pointes 
CATEGORY 
DRUG 
Anti-arrhythmic agents 
Amiodarone 
Disopyramide 
Dofetilide 
Dronedarone 
Flecainide 
Ibutilide 
Procainamide 
Quinidine 
Sotalol 
 
Anti-infective agents 
Azithromycin 
Ciprofloxacin 
Clarithromycin 
Erythromycin 
Levofloxacin 
Moxifloxacin 
 
Anti-malarial agents 
Chloroquine 
 
Anti-psychotic agents 
Chlorpromazine 
Droperidol 
Haloperidol 
Pimozide 
Thioridazine 
 
Anesthetics, General 
Propofol 
Sevoflurane 
 
Others 
Methadone 
Papaverine HCl 
 
Source: https://crediblemeds.org/ (accessed 19 December 2017) and Reference 17