A Double Blind, Randomized, Placebo-Controlled, Phase 1 Dose Escalation Trial to Evaluate the Safety and Immunogenicity of an Inactivated Yellow Fever Virus Vaccine, HydroVax-002 YFV, in Healthy Adults
Phase: PHASE1
Conditions: Yellow Fever
Sponsor: Najit Technologies, Inc.

 
 
 
 
 
 
 
 
 
 
Complete Clinical Protocol 
 
A Double Blind, Randomized, Placebo-Controlled, Phase 1 
Dose Escalation Trial to Evaluate the Safety and 
Immunogenicity of an Inactivated Yellow Fever Virus Vaccine, 
HydroVax-002 YFV, in Healthy Adults 
 
NCT05172544 
 
24 January, 2023 

NTI Protocol 20-001 
 
 
 
 
 
 
 
 
 
 
Version 6.0 
HydroVax-002 YFV Phase 1 Study  
 
 
 
 
 
 
 
 
24 January 2023 
 
 
 
Complete Clinical Protocol 
Proprietary and Confidential 
 
Page 1 of 84 
 
 
 
 
 
 
 
A Double Blind, Randomized, Placebo-Controlled, Phase 1 Dose Escalation 
Trial to Evaluate the Safety and Immunogenicity of an Inactivated Yellow 
Fever Virus Vaccine, HydroVax-002 YFV, in Healthy Adults 
 
Complete Clinical Protocol 
 
 
 
Protocol Number:  20-001 
IND Sponsor: Najít Technologies, Inc. (NTI) 
Version Number: 6.0 
 
 
 

NTI Protocol 20-001 
 
 
 
 
 
 
 
 
 
 
Version 6.0 
HydroVax-002 YFV Phase 1 Study  
 
 
 
 
 
 
 
 
24 January 2023 
 
 
 
Complete Clinical Protocol 
Proprietary and Confidential 
 
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TABLE OF CONTENTS 
TITLE PAGE 
1 
STATEMENT OF COMPLIANCE 
2 
TABLE OF CONTENTS 
3 
LIST OF FIGURES 
8 
LIST OF TABLES 
8 
 
PROTOCOL SUMMARY 
9 
1.1 
Synopsis 
9 
 Study overview 
9 
 Enrollment Period 
9 
 General 
9 
 Study Population 
10 
 Inclusion Criteria 
10 
 Exclusion Criteria 
11 
 Study Intervention 
13 
 Staged Enrollment 
13 
Table 1. Study design and dose escalation 
14 
1.2 
Schedule of Assessments (SoA) 
15 
Table 2. Schedule of Assessments  (SoA) 
15 
 
INTRODUCTION 
18 
2.1 
Study Rationale 
18 
2.2 
Background 
18 
 Purpose of Study 
18 
2.2.1.1 Yellow Fever Virus 
18 
2.2.1.2 Candidate Study Vaccine – HydroVax-002 YFV 
19 
Figure 1. Convalescent serum from HydroVax-002 YFV vaccinated mice protects against lethal 
challenge. 
20 
Figure 2. HydroVax-002 YFV vaccination induces robust virus-specific neutralizing antibody responses 
and protective immunity against lethal yellow fever. 
22 
2.3 
Risk/Benefit Assessment 
23 
 Known Potential Risks 
23 
 Known Potential Benefits 
24 
 Assessment of Potential Risks and Benefits 
24 
 
STUDY OBJECTIVES AND OUTCOME MEASURES 
25 
 
STUDY DESIGN 
27 
4.1 
Overall Design 
27 
Table 3. Laboratory samples and estimated blood volume (mL) by visit 
28 
4.2 
Scientific Rationale for Study Design 
28 
 Inclusion of Placebo 
28 
4.3 
Justification for Dose 
28 
 
STUDY POPULATION 
30 
5.1 
Inclusion Criteria 
30 

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5.2 
Exclusion Criteria 
32 
5.3 
Exclusion of Specific Populations 
33 
5.4 
Inclusion of Vulnerable Participants 
34 
5.5 
Lifestyle Considerations 
34 
5.6 
Screen Failures 
34 
5.7 
Strategies for Recruitment and Retention 
34 
 Recruitment 
34 
 Retention 
35 
 Compensation Plan for Subjects 
35 
 Costs 
35 
 
STUDY PRODUCT 
36 
6.1 
Study Product and Administration 
36 
 Study Product Description 
36 
6.1.1.1 Vaccine 
36 
6.1.1.2 Placebo and Diluent 
36 
 Dosing and Administration 
36 
 Dose Escalation 
36 
 Dose Modifications 
36 
 Overdosage 
37 
6.2 
6.2 Preparation/Handling/Storage/Accountability 
37 
 Acquisition and Accountability 
37 
 Formulation, Appearance, Packaging, and Labeling 
37 
 Product Storage and Stability 
38 
 Preparation 
38 
6.3 
Measures to Minimize Bias: Randomization and Blinding 
38 
 Treatment Assignment Procedures 
38 
 Randomization 
38 
 Blinding and Masking Procedures 
39 
6.3.3.1 Unblinding 
39 
6.4 
Study Intervention Compliance 
39 
6.5 
Concomitant Therapy 
39 
 Rescue Medicine 
40 
 
STUDY INTERVENTION DISCONTINUATION AND PARTICIPANT 
DISCONTINUATION/WITHDRAWAL 
41 
7.1 
Halting Criteria and Discontinuation of Study Intervention 
41 
 Study Halting Criteria 
41 
 Individual Halting Criteria 
42 
7.2 
Participant Withdrawal from the Study and Replacement 
43 
 Participant Withdrawal 
43 
 Subject Replacement 
43 
7.3 
Lost to Follow-Up 
44 
7.4 
Follow up for subjects that discontinued study intervention 
44 
 
STUDY ASSESSMENTS AND PROCEDURES 
45 

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8.1 
Screening and Immunogenicity Assessments 
45 
 Screening Procedures 
45 
 Immunogenicity Assessments 
46 
8.1.2.1 Humoral Immunogenicity Tests 
46 
8.1.2.2 Specimen Preparation, Handling, and Shipping 
46 
8.2 
Safety and Other Assessments 
46 
 Physical examination 
46 
 Safety Clinical Laboratory Tests 
46 
 Viremia Detection Tests 
47 
 Additional Sera Samples 
47 
8.3 
Procedures to be Followed in the Event of Abnormal Laboratory Test Values or Abnormal Clinical 
Findings 
47 
8.4 
Unscheduled Visit 
47 
8.5 
Adverse Events and Serious Adverse Events 
47 
 Definition of Adverse Events (AE) 
47 
8.5.1.1 Solicited Adverse Events/Reactogenicity 
48 
Table 4. Table of injection site symptom grading 
49 
Table 5. Table of erythema/redness and induration/swelling grading 
49 
Table 6. Table of systemic symptom grading 
49 
Table 7. Table of fever grading 
50 
8.5.1.2 Unsolicited Adverse Events 
50 
8.5.1.3 Unexpected 
50 
 Definition of Serious Adverse Events (SAE) 
51 
8.5.2.1 Suspected Unexpected Serious Adverse Reactions (SUSAR) 
51 
 Classification of an Adverse Events 
51 
8.5.3.1 Seriousness and Severity of Event 
52 
8.5.3.2 Relationship to Study Intervention 
52 
 Time Period and Frequency for Event Assessment and Follow-up 
52 
 Adverse Event Reporting 
53 
8.5.5.1 Investigators Reporting of AEs 
53 
 Serious Adverse Event Reporting 
53 
8.5.6.1 Investigators Reporting of AEs 
53 
8.5.6.2 Regulatory Reporting of AEs 
53 
 Adverse Events of Special Interest 
54 
 Reporting of Pregnancy 
54 
8.6 
Unanticipated Problems 
54 
 Definition of Unanticipated Problems (UP) 
54 
 Unanticipated Problem Reporting 
54 
 Reporting Unanticipated Problems to Participants 
55 
 
STATISTICAL CONSIDERATIONS 
56 
9.1 
Study Hypotheses 
56 
9.2 
Sample Size Considerations 
56 
 Safety 
56 

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Table 8. Probability of observing one or more adverse events in one dosage group (n=10) given 
particular true event rates 
56 
 Immunogenicity 
57 
9.3 
Populations for Analyses 
57 
9.4 
Statistical Analyses 
57 
 General Approach 
57 
 Analysis of Primary Outcome Measures (Safety) 
57 
 Analysis of Secondary and Exploratory Outcome Measures (Immunogenicity) 
58 
 Timing of Analyses 
59 
9.4.4.1 Interim Safety Analyses 
59 
9.4.4.2 Final Analyses 
59 
 Sub-group Analyses 
60 
 Tabulation of Individual participant Data 
60 
 SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS 
61 
10.1 
Regulatory, Ethical and Study Oversight Considerations 
61 
 Informed Consent Process 
61 
10.1.1.1 Requirements for Permission by Parents/Guardians and Assent by Children (in case of a minor) 
62 
10.1.1.2 Other Informed Consent Procedures 
63 
10.1.1.3 Ethical Standard 
63 
10.1.1.4 Institutional Review Board 
63 
 Study Termination and Closure 
63 
 Confidentiality and Privacy 
64 
 Future Use of Stored Specimens for Secondary Research 
65 
 Key Roles and Study Governance 
65 
Table 9. Table of Key Roles 
65 
 Safety Oversight 
66 
10.1.6.1 Safety Monitoring Committee (SMC) 
66 
10.1.6.2 Internal Safety Review Committee (ISRC) 
67 
 Clinical Monitoring/Site Monitoring Plan 
67 
 Quality Control and Quality Assurance 
68 
 Data Handling and Record Keeping 
68 
10.1.9.1 Data Collection and Management Responsibilities 
69 
10.1.9.2 Study Records Retention 
69 
10.1.9.3 Source Records 
69 
 Protocol Deviations 
70 
 Publication and Data Sharing 
70 
10.1.11.1 
Human Data Sharing Plan 
70 
10.1.11.2 
Genomic Data Sharing Plan 
70 
10.1.11.3 
Publication 
71 
 Conflict of Interest Policy 
71 
10.2 
Additional Considerations 
72 
 Research Related Injuries 
72 
10.3 
Abbreviations 
73 
10.4 
Protocol Amendment History 
76 
Table 10. Protocol Amendment History 
76 

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 REFERENCES 
79 
APPENDIX A 
81 
Table 11. Hematology toxicity table 
82 
Table 12. Blood chemistries toxicity table 
82 
Table 13. Liver enzymes toxicity table 
83 
Table 14. Urine dipstick toxicity table 
83 
Table 15. Cardiovascular toxicity table 
84 
Table 16. Respiratory toxicity table 
84 

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LIST OF FIGURES 
 
Figure 1. Convalescent serum from HydroVax-002 YFV vaccinated mice protects against lethal challenge. 20 
Figure 2. HydroVax-002 YFV vaccination induces robust virus-specific neutralizing antibody responses and 
protective immunity against lethal yellow fever. ............................................................................. 22 
 
 
LIST OF TABLES 
 
Table 1. Study design and dose escalation ....................................................................................................... 14 
Table 2. Schedule of Assessments  (SoA) ....................................................................................................... 15 
Table 3. Laboratory samples and estimated blood volume (mL) by visit ........................................................ 28 
Table 4. Table of injection site symptom grading ........................................................................................... 49 
Table 5. Table of erythema/redness and induration/swelling grading ............................................................. 49 
Table 6. Table of systemic symptom grading .................................................................................................. 49 
Table 7. Table of fever grading ........................................................................................................................ 50 
Table 8. Probability of observing one or more adverse events in one dosage group (n=10) given particular 
true event rates .................................................................................................................................. 56 
Table 9. Table of Key Roles ............................................................................................................................ 65 
Table 10. Protocol Amendment History .......................................................................................................... 76 
Table 11. Hematology toxicity table ................................................................................................................ 82 
Table 12. Blood chemistries toxicity table ....................................................................................................... 82 
Table 13. Liver enzymes toxicity table ............................................................................................................ 83 
Table 14. Urine dipstick toxicity table ............................................................................................................. 83 
Table 15. Cardiovascular toxicity table ........................................................................................................... 84 
Table 16. Respiratory toxicity table ................................................................................................................. 84 
 
 
 

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 PROTOCOL SUMMARY 
1.1 
Synopsis 
 
Study overview 
This study will evaluate the safety and immunogenicity of two different dosages of a novel peroxide-inactivated 
whole-virus yellow fever vaccine, HydroVax-002 YFV, in healthy adults.  The currently available yellow fever 
vaccine is the live, attenuated YFV-17D vaccine. This live attenuated vaccine is contraindicated in multiple 
vulnerable populations including infants <9 months of age and immunosuppressed individuals such as those with 
acquired immunodeficiency syndrome, thymic disease, or persons with hypersensitivity to eggs. Special 
precautions are also noted for pregnant women, nursing mothers and adults over the age of 60 [1]. These 
contraindications and precautions stem from known serious adverse events associated with the vaccine including 
a serious nervous system/neurotropic condition termed yellow fever associated neurotropic disease (YEL-AND).  
In addition to YEL-AND, vaccination with live YFV-17D can cause severe multiple organ system failure, 
described as yellow fever vaccine-associated viscerotropic disease (YEL-AVD), a syndrome closely resembling 
wild-type yellow fever virus (YFV).  Because HydroVax-002 YFV is based on inactivated virus that is unable to 
replicate or spread to the brain or nervous system, we anticipate that these vaccine complications will be 
eliminated.  
 
 
Enrollment Period 
It is anticipated that enrollment may be completed in 3.5-4 months. 
 
 
General 
This trial will be a randomized, placebo controlled, double-blind (within dosing group), dose escalation Phase 1 
trial evaluating dosages of 1 mcg and 5 mcg of HydroVax-002 YFV vaccine given intramuscularly on Day 1 (the 
day of first vaccination is defined as Day 1) and Day 29 in healthy adults ≥18 and <50 years of age. The study 
will consist of two dosing groups of HydroVax-002 YFV vaccine to be enrolled sequentially. Each dose group 
will consist of 10 individuals who receive HydroVax-002 YFV and 2 or 3 who receive placebo. The initial dose 
of HydroVax-002 YFV to be evaluated in Group 1 will be 1 mcg and the next dose to be evaluated in Group 2 
will be 5 mcg. Following assessment of safety and reactogenicity data of Group 1 by the Safety Monitoring 
Committee (SMC), the vaccine dose will be increased to 5 mcg for Group 2.  All participants will be monitored 
for local and systemic solicited adverse events for 7 days following each vaccination, unsolicited adverse events 
for 28 days following each vaccination, and for the occurrence of new onset chronic medical conditions and 
serious adverse events from the time of first vaccination until 6 months following the second vaccination.  Safety 
laboratory tests will be performed prior to vaccination and at days 4 and 15 following vaccination.  YFV 
neutralizing assays will be performed on blood samples collected prior to each vaccine, 15 days following each 
vaccine, and at days 29, 57, and 180 following the second dose of vaccine.  
 
For each dosing group, the first four subjects enrolled (the sentinel subgroup) will include three HydroVax-002 
YFV recipients and one placebo recipient.  After the four subjects in the Group 1 sentinel subgroup are enrolled 
and given their first vaccination, enrollment will then be stopped pending a review of the clinical laboratory, 
reactogenicity, and safety data collected through the post vaccination Day 8 visit for the last of those subjects. 

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This review may be conducted by an internal safety review committee (ISRC), consisting of the Blinded medical 
monitor, clinical program manager, and the PI, or by the SMC, as indicated and detailed in Section 10.1.6. 
Approval by the reviewing group will allow administration of the second vaccination to the sentinel subgroup 
and continued enrollment of the remaining 9 Group 1 subjects (expanded group) to resume to complete enrollment 
of 13 participants (10 vaccine and 3 placebo).  
 
After Group 1 enrollment is completed enrollment will be stopped pending an SMC review of the clinical 
laboratory, reactogenicity, and adverse event information through the post 2nd vaccination Day 15 visit for all 
Group 1 subjects. Approval by the SMC will allow escalation to Group 2, and initiation of enrollment of the 
Group 2 sentinel subgroup.  After the four subjects in the Group 2 sentinel subgroup are enrolled and given their 
first vaccination, enrollment will then be stopped pending a safety review of the sentinel subgroup as specified 
for the low dose cohort. Approval will allow administration of vaccination 2 to the sentinel subgroup and 
continued enrollment of Group 2 subjects (expanded group) to complete study enrollment. 
 
This study will accrue 25 participants: (10 receiving 1 mcg of HydroVax-002 YFV; 10 receiving 5 mcg of 
HydroVax-002 YFV; and 5 receiving placebo). 
 
 
Study Population 
Healthy, male and female adults ≥18 and <50 years of age 
 
 
Inclusion Criteria 
1. Provide written informed consent prior to initiation of any study procedures. 
2. Are able to understand and comply with planned study procedures and be available for all study visits. 
3. Must agree to the collection of venous blood per protocol. 
4. Are males or non-pregnant females, ≥18 and <50 years of age, inclusive at time of enrollment. 
5. Are in good health1. 
1As determined by medical history and physical examination to evaluate acute or currently ongoing chronic medical or 
psychiatric diagnoses or conditions, defined as those that have been present for at least 90 days, which would affect the 
assessment of the safety of subjects or the immunogenicity of study vaccinations. Chronic medical diagnoses or conditions 
should be stable for the last 60 days (no hospitalizations, emergency room or urgent care for condition, or invasive medical 
procedure and no adverse symptoms that need medical intervention such as medication change/supplemental oxygen). This 
includes no change in chronic prescription medication, dose or in the 60 days prior to enrollment. Any prescription change 
that is due to change of health care provider, insurance company, etc., or that is done for financial reasons, as long as in the 
same class of medication, will not be considered a deviation of this inclusion criterion. Subjects may be on chronic or as needed 
(prn) medications if, in the opinion of the site PI or appropriate sub-investigator, they pose no additional risk to subject safety 
or assessment of reactogenicity and immunogenicity and do not indicate a worsening or treatment of continued symptoms of 
medical diagnosis or condition. Note: Low dose topical, corticosteroids as outlined in the Subject Exclusion Criteria (see 
Section 5.2) as well as herbals, vitamins and supplements are permitted. 
6. Oral temperature is less than 100.0℉. 
7. Pulse is 47 to 100 beats per minute, inclusive. 
8. Systolic blood pressure is 85 to 140 mmHg, inclusive. 

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9. Diastolic blood pressure is 55 to 90 mmHg, inclusive. 
10. Screening laboratories (WBC, Hgb, PLTs, Sodium, Potassium, Bicarbonate, Calcium, Cr, non-fasting 
glucose, ALT, AST, TBIL and urine protein and glucose) are within acceptable parameters2. 
          2Hematology, blood chemistry and liver enzymes must be Grade 1 or less at screening (Refer to Table 11, Table 12, and Table 
13, respectively); urine glucose negative and urine protein no greater than trace at screening for subjects to qualify for 
randomization and vaccination. 
11. Negative test for HIV, hepatitis B virus (HBV), and hepatitis C virus (HCV) at screening blood draw 
12. Women of childbearing potential3 must use an acceptable contraception method4 from at least 30 days 
before the first study vaccination until 30 days after the second study vaccination 
3Not sterilized via, bilateral oophorectomy, salpingectomy, hysterectomy, or successful Essure® placement (permanent, non-
surgical, non-hormonal sterilization) with documented radiological confirmation test at least 90 days after the procedure, and 
still menstruating or <1 year has passed since the last menses if menopausal. 
4Includes non-male sexual relationships, full abstinence from sexual intercourse with a male partner, monogamous relationship 
with vasectomized partner who has been vasectomized for 180 days or more and shown to be azoospermic prior to the subject 
receiving the study vaccination, effective intrauterine devices, NuvaRing®, tubal ligation, and licensed hormonal methods such 
as implants, injectables or oral contraceptives (i.e. “the pill”). 
13. Women of childbearing potential must have a negative serum pregnancy test at screening and a negative 
urine pregnancy test within 24 hours prior to each study vaccination. 
14. Sexually active males must agree to use a medically acceptable form of contraception5 in order to be in 
this study and must agree to continue such use until day 30 after the last vaccination.  
5Medically acceptable contraceptives include: (1) surgical sterilization (such as a vasectomy), or (2) a condom used with a 
spermicide.  Contraceptive measures such as Plan B™, sold for emergency use after unprotected sex, are not acceptable 
methods for routine use. 
 
 
Exclusion Criteria 
1. Have an acute illness1 or acute febrile illness (oral temperature  ≥ 38℃ [100.4℉]), as determined by the 
site PI or appropriate sub-investigator, within 72 hours prior to study vaccination. 
1An acute illness which is nearly resolved with only minor residual symptoms remaining is allowable if, in the opinion of the 
site PI or appropriate sub-investigator, the residual symptoms will not interfere with the ability to assess safety parameters as 
required by the protocol. 
2. Have any medical disease or condition that, in the opinion of the site PI or appropriate sub-investigator, 
is a contraindication to study participation2. 
2Including acute, subacute, intermittent or chronic medical disease or condition that would place the subject at an unacceptable 
risk of injury, render the subject unable to meet the requirements of the protocol, or may interfere with the evaluation of 
responses or the subject’s successful completion of this trial. 
3. Have immunosuppression as a result of an underlying illness or treatment, a recent history or current use 
of immunosuppressive or immunomodulating disease therapy. 
4. Use of anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study vaccination. 
5. Have known active or recently active (12 months) neoplastic disease or a history of any hematologic 
malignancy. Non-melanoma, treated, skin cancers are permitted. 
6. Known allergy to components of the study product3. 
3Including the following: aluminum hydroxide, sorbitol, potassium chloride, sodium chloride and polysorbate80 (Tween80) 

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7. Unstable seizure disorder (defined as requiring medication for seizure control or with seizure activity 
within the past 3 years). 
8. Have a history of alcohol or drug abuse within 5 years prior to study vaccination. 
9. Have any diagnosis, current or past, of schizophrenia, bipolar disease or other psychiatric diagnosis that 
may interfere4 with subject compliance or safety evaluations. 
4As determined by the site PI or appropriate sub-investigator. 
10. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement for danger to self 
or others within 5 years prior to study vaccination. 
11. Have a history of asthma, other than mild, well-controlled asthma5 
5Cold or exercise induced asthma controlled with inhaled medications other than inhaled corticosteroids is permissible. 
Participants should be excluded if they require daily bronchodilator use, or have had an asthma exacerbation requiring 
oral/parenteral steroid use or have used theophylline or inhaled corticosteroids in the past year. 
12. Have a history of diabetes mellitus. 
13. Have taken oral or parenteral (including intra-articular) or chronic topical corticosteroids of any dose 
within 30 days prior to study vaccination6. 
6Corticosteroid nasal sprays for allergic rhinitis are permissible. Persons using a topical corticosteroid for a limited duration 
for mild uncomplicated dermatitis such as poison ivy or contact dermatitis may be enrolled the day after their therapy is 
completed. 
14. Have taken high-dose inhaled corticosteroids7 within 30 days prior to study vaccination. 
7High-dose defined as per age as using inhaled high-dose per reference chart in the National Heart, Lung and Blood Institute 
Guidelines for the Diagnosis and Management of Asthma (EPR-3) or other lists published in UPTODATE. 
15. Received or plan to receive a licensed, live vaccine within 30 days before or after each study vaccination. 
16. Received or plan to receive a licensed, inactivated vaccine or allergy desensitization shot within 14 days 
before or after each study vaccination. 
17. Received an experimental agent8 within 30 days prior to the study vaccination or expect to receive another 
experimental agent9 during the trial-reporting period10. 
8Including vaccine, drug, biologic, device, blood product, or medication. 
9Other than from participation in this trial. 
10Approximately 7 months after the first study vaccination. 
18. Are participating or plan to participate in another clinical trial with an interventional agent11 that will be 
received during the trial-reporting period12. 
11Including licensed or unlicensed vaccine, drug, biologic, device, blood product, or medication. 
12Approximately 7 months after the first study vaccination. 
19. Female subjects who are breastfeeding or plan to breastfeed from the time of the first study vaccination 
through 30 days after the last study vaccination. 
20. Receipt of blood products or immunoglobulin within six months prior to enrollment. 
21. Donation of a unit of blood within 60 days prior to enrollment or intends to donate blood during the study 
period. 
22. Body mass index (BMI) ≥ 35. 

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23. History of a visit to South America, sub-Saharan Africa or Southeast Asia lasting one month or more. 
24. Planned travel to areas known to be endemic with Yellow Fever virus during the study period. 
25. History of military service. 
26. History of vaccination against dengue, yellow fever, tick-borne encephalitis, or Japanese encephalitis. 
27. History of vaccination with other flavivirus candidate vaccines. 
28. Attended primary (grade) school in Austria, Germany, Japan, South Korea, India, Thailand, Nepal, 
Vietnam, or Taiwan (locations where the tick-borne encephalitis vaccine is given). 
29. History of yellow fever. 
30. Plan to have a major change in exercise routine or perform strenuous exercise from 72 hours before any 
dose of study vaccine/placebo and for 72 hours before any safety laboratories13.  
13Safety laboratories are obtained on Days 4 and 15 following each dose of study product. 
31. Contraindication to intramuscular vaccination of either upper arm (for example, due to lymphadenectomy 
or obscuring tattoos). 
 
 
 
Study Intervention 
Investigational Vaccine: HydroVax-002 YFV Yellow Fever Virus (YFV), inactivated vaccine, alum-adsorbed, 
administered intramuscularly in a two-dose series on Day 1 and Day 29 at a dose of 1 mcg and at a dose of 5 mcg.   
Controls will receive sterile 0.9% NaCl placebo intramuscularly at days 1 and 29.  
 
 
Staged Enrollment 
For each dosing group, the first four subjects enrolled (the sentinel subgroup) will include three HydroVax-002 
YFV recipients and one placebo recipient (Table 1).  After the four subjects in the Group 1 sentinel subgroup are 
enrolled and given their first vaccination, enrollment will then be stopped pending a review of the clinical 
laboratory, reactogenicity, and safety data collected through the post vaccination Day 8 visit for the last of those 
subjects. This review may be conducted by an internal safety review committee (ISRC), consisting of the Blinded 
medical monitor, clinical program manager, and the PI, or by the SMC, as indicated and detailed in Section 10.1.6. 
Approval by the reviewing group will allow administration of the second vaccination to the sentinel subgroup 
and continued enrollment of the remaining 9 Group 1 subjects (expanded group) to resume to complete enrollment 
of 13 participants (10 vaccine and 3 placebo).  
 
After Group 1 enrollment is completed enrollment will be stopped pending an SMC review of the clinical 
laboratory, reactogenicity, and adverse event information through the post 2nd vaccination Day 8 visit for all 
Group 1 subjects. Approval by the SMC will allow escalation to Group 2, and initiation of enrollment of the 
Group 2 sentinel subgroup.  After the four subjects in the Group 2 sentinel subgroup are enrolled and given their 
first vaccination, enrollment will then be stopped pending a safety review of the sentinel subgroup as specified 
for the low dose cohort. Approval will allow administration of vaccination 2 to the sentinel subgroup and 
continued enrollment of Group 2 subjects (expanded group) to complete study enrollment. 
 

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 INTRODUCTION 
2.1 
Study Rationale 
Yellow fever virus (YFV) is a mosquito-borne pathogen that is endemic in over 40 countries [2] and clinical 
presentation often involves severe acute onset with fever, nausea, vomiting, hepatitis, hemorrhage and renal 
failure.  The case fatality rate for severe cases of yellow fever ranges from 20-60% [3, 4].  Prevention or reduction 
of disease burden is mainly accomplished through immunization with a live-attenuated yellow fever virus (e.g., 
YFV-17D or YFV-17DD) but these live vaccines are specifically contraindicated in young infants, pregnant or 
nursing mothers of young infants, people with egg allergies or people with a number of pre-existing medical 
conditions.  In addition, specific warnings are in place for individuals >60 years of age due to the risk of rare, but 
potentially life-threatening, complications.  This means that there is currently no safe vaccine available to protect 
these vulnerable populations.  To address this unmet clinical need, we have developed an advanced yellow fever 
vaccine formulation in which the virus is inactivated using the proprietary hydrogen peroxide-based HydroVax™ 
technology. 
 
2.2 
Background 
 
Purpose of Study 
The Sponsor proposes a Phase 1 double-blind placebo-controlled clinical trial for a first-in-person evaluation of 
the safety and immunogenicity of a novel peroxide-inactivated whole-virus yellow fever vaccine. 
 
2.2.1.1 
Yellow Fever Virus 
Yellow fever virus (YFV) represents an important mosquito-borne human pathogen that is endemic in more than 
40 countries in sub-Saharan Africa and South America [2].  Although disease may be mild in some instances, 
clinical presentation often involves severe acute onset with fever, nausea, vomiting, hepatitis, hemorrhage and 
renal failure.  The case fatality rate for severe cases of yellow fever ranges from 20-60% [3, 4].  There are no 
licensed antiviral drugs to treat yellow fever and prevention or reduction of disease burden is mainly accomplished 
through vaccination as well as through vector control measures.  Though the requirements for protective vaccine-
mediated immunity are unknown for many viruses, a correlate of immunity exists for YFV.  Protective immunity 
against YFV was originally defined as a log neutralizing index (LNI) of ≥0.7 [5, 6] based on NHP challenge 
studies but more  recent yellow fever vaccine clinical trials have been updated to use more reliable serum-dilution, 
constant-virus plaque-reduction neutralization tests (PRNT) [7, 8]. 
 
The live, attenuated YFV-17D vaccine was developed in 1936 and although there are two main substrains that 
are used in commercial manufacturing (17D and 17DD), there appear to be no major differences in safety or 
immunogenicity between YFV-17D vaccines [3, 9-11].  Yellow fever vaccination is recommended for people 
living in endemic areas and for travelers who may visit endemic areas, and for many years, YFV-17D has been 
considered “one of the safest vaccines in the world” [12].  However, this live attenuated vaccine is contraindicated 
in persons with hypersensitivity to eggs as well as several vulnerable populations including infants <9 months of 
age and immunosuppressed individuals such as those with acquired immunodeficiency syndrome, leukemia, 
lymphoma, thymic disease, generalized malignancy, and patients who are undergoing drug therapy (e.g., systemic 
corticosteroids, alkylating drugs, antimetabolites or other immunomodulatory drugs) or radiation therapy. Thymic 

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2.3 
Risk/Benefit Assessment 
 
Known Potential Risks 
This is the first human trial of HydroVax-002 YFV.  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Common potential side effects resulting from intramuscular injections include stinging, discomfort, redness of 
skin, or mild bruising at vaccine injection site.  
 
Uncommon side effects include general signs and symptoms associated with administration of the vaccine 
injection, including fever, chills, rash, aches and pains, nausea, dizziness and fatigue. These side effects will be 
monitored, but are generally short term, mild to moderate severity and usually do not require treatment. 
 
Rare risks with any injection procedure include infection at the site of injection. Signs of infection at the injection 
site include: severe pain, erythema, induration, warmth or drainage. There may be side effects from the study 
products, which may be serious or life threatening that we do not know about yet. 

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Known Potential Benefits 
Participation in this study provides no known direct benefit to the individual subjects.  
 
 
Assessment of Potential Risks and Benefits 
This study may not benefit participants directly. As this is a first-in-person trial, it is unknown whether the study 
product, HydroVax-002 YFV, will help to protect subjects from yellow fever disease or, if it does, how long that 
protection may last.  The study placebo, saline solution, would offer no known benefit to study participants.   As 
detailed in Section 2.3.1, there are potential risks for any intramuscular injection. 
 
Study risk is minimized by using a dose escalation structure, as well as a sentinel group approach within each 
dose level.  Group safety reviews occur between sentinel and expanded groups, and between dose groups.  In 
addition, each individual study subject is followed closely during acute time periods following test article 
administration. 
 
While individual subjects may not benefit from involvement in this study, their participation is crucial to the 
larger societal benefit of improved medical interventions.  As noted in Section 2.1, the current live attenuated 
YFV vaccine is contraindicated in multiple vulnerable populations including infants <9 months of age and 
immunosuppressed individuals such as those with acquired immunodeficiency syndrome, thymic disease, or 
persons with hypersensitivity to eggs.  By participating in this study, subjects are enabling the development of a 
potential advancement to address this current unmet medical need. 
 
 
 

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To assess long-term YFV-specific neutralizing 
antibody responses after a second dose of HydroVax-
002 YFV vaccine given at dose levels of 1 mcg and 5 
mcg 
• Geometric mean neutralizing titers at Day 180 
following second vaccination  
• Reverse 
cumulative 
distribution 
curve 
of 
neutralizing titers on Day 180 after the second 
vaccination for each dose group and for all dose 
groups combined 

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 STUDY DESIGN 
4.1 
Overall Design 
This trial will be a randomized, placebo controlled, double-blind (within dosing group), dose escalation Phase 1 
trial evaluating dosages of 1 mcg and 5 mcg of HydroVax-002 YFV vaccine given intramuscularly on Day 1 (the 
day of first vaccination is defined as Day 1) and Day 29 in healthy adults ≥18 and <50 years of age. The study 
will consist of two dosing groups of HydroVax-002 YFV vaccine to be enrolled sequentially. Each dose group 
will consist of 10 individuals who receive HydroVax-002 YFV, as well as 5 total subjects who receive placebo. 
Each dose-group will include a sentinel subgroup consisting of 3 vaccine and 1 placebo recipient. In each of the 
two (1 mcg and 5 mcg) dose phases, enrollment is halted after the dose 1 vaccination of the sentinel subgroup. 
Following assessment of safety and reactogenicity data of Group 1 by the Safety Monitoring Committee (SMC), 
the vaccine dose will be increased to 5 mcg for Group 2.  
 
For each dosing group, the first 4 subjects enrolled (the sentinel subgroup) will include 3 HydroVax-002 YFV 
recipients and 1 placebo recipient.  After the 4 subjects in the Group 1 sentinel subgroup are enrolled and given 
their first vaccination, enrollment will then be stopped pending a review of the clinical laboratory, reactogenicity, 
and safety data collected through the post vaccination one Day 8 visit for the last of those subjects. This review 
will be conducted by an internal safety review committee (ISRC), consisting of the Blinded medical monitor, 
clinical program manager and the PI.  If deemed necessary by the ISRC, a further review will be conducted by 
the SMC, as detailed in Section 10.1.6.  Approval by the reviewing group will allow administration of the second 
vaccination to the sentinel subgroup and continued enrollment of the remaining 9 Group 1 subjects (expanded 
group) to resume to complete enrollment of 13 participants (10 vaccine and 3 placebo).  
 
After Group 1 enrollment is completed enrollment will be stopped pending an SMC review of the clinical 
laboratory, reactogenicity, and adverse event information through the post 2nd  vaccination Day 15 visit for all 
Group 1 subjects. Approval by the SMC will allow escalation to Group 2, and initiation of enrollment of the 
Group 2 sentinel subgroup.  After the 4 subjects in the Group 2 sentinel subgroup are enrolled and given their 
first vaccination, enrollment will then be stopped pending a safety review of the sentinel subgroup as specified 
for the low dose cohort. Approval will allow administration of vaccination 2 to the sentinel subgroup and 
continued enrollment of Group 2 subjects (expanded group) to complete study enrollment.  
 
All subjects will complete a subject memory aid for 7 days after each study vaccination. Unsolicited, non-serious 
adverse events and serious adverse events will be collected through 29 days after the second vaccination. Serious 
adverse events and new onset of chronic medical conditions will be collected from the time of vaccination 
throughout the study period. Subjects’ participation will be approximately 8 months. Table 2 provides a detailed 
outline of the study schedule. 
 
Blood samples will be obtained at specified time points following each vaccination for safety laboratory 
assessments and assessment of humoral immune responses.  
 
 
Table 3 provides the estimated blood volume of collection at each visit and cumulatively throughout the study. 
 

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formulated with 0.1% aluminum hydroxide (Al (OH)3)]. The 3-dose vaccination schedule represents n+1 for the 
proposed 2-dose schedule to be administered during this Phase 1 clinical trial. This dose of vaccine was well-
tolerated in the study animals. These results indicate that the vaccine test article demonstrates an acceptable pre-
clinical safety profile at the planned maximum dose level of 5 mcg. 
 
This route and vaccination schedule is commonly used for other inactivated whole-virus vaccines formulated with 
alum.  Analysis of antigen dose used in other inactivated alum-adsorbed, whole-virus vaccines consisting of 
closely related flaviviruses produced in Vero cells provides information of potential relevance for this dose 
ranging study of HydroVax-002 YFV.  The commercial tick-borne encephalitis virus vaccine, TicoVac, contains 
5 mcg/dose, and the commercial Japanese encephalitis virus vaccine, IXIARO, contains 6 mcg/dose.  The 
experimental yellow fever virus vaccine, XRX-001, was tested in Phase I trials at 4.8 mcg/dose and at 0.48 
mcg/dose.[8]  Antiviral immunity following a single dose of inactivated Japanese encephalitis virus vaccine is 
low after a single 6 mcg dose,[35] whereas two or three intramuscular injections of a 12 mcg/dose vaccine were 
tested and found to be safe, well tolerated, and immunogenic [36]. 
 
Based on prior results with a closely related inactivated flavivirus vaccine (HydroVax-001 WNV, [37]) we do not 
anticipate significant safety signals.  Based on the animal model evaluations of HydroVax-002 YFV and studies 
of other inactivated alum-absorbed flavivirus vaccines, the antigen doses to be evaluated in this Phase 1 trial of 
HydroVax are 1 mcg and 5 mcg. 
 
 
 

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 STUDY POPULATION 
Only subjects who meet the inclusion criteria and do not meet the exclusion criteria will be eligible for enrollment. 
Inclusion and exclusion criteria must be assessed by a study clinician licensed to make medical diagnoses and 
listed on the Form FDA 1572 as the site principal investigator or sub-investigator. No exemptions are granted to 
inclusion and exclusion criteria in studies sponsored by NTI.  
 
5.1 
Inclusion Criteria 
1. Provide written informed consent prior to initiation of any study procedures. 
2. Are able to understand and comply with planned study procedures and be available for all study visits. 
3. Must agree to the collection of venous blood per protocol. 
4. Are males or non-pregnant females, ≥18 and <50 years of age, inclusive at time of enrollment. 
5. Are in good health1. 
1As determined by medical history and physical examination to evaluate acute or currently ongoing chronic medical or 
psychiatric diagnoses or conditions, defined as those that have been present for at least 90 days, which would affect the 
assessment of the safety of subjects or the immunogenicity of study vaccinations. Chronic medical diagnoses or conditions 
should be stable for the last 60 days (no hospitalizations, emergency room or urgent care for condition, or invasive medical 
procedure and no adverse symptoms that need medical intervention such as medication change/supplemental oxygen). This 
includes no change in chronic prescription medication, dose or in the 60 days prior to enrollment. Any prescription change 
that is due to change of health care provider, insurance company, etc., or that is done for financial reasons, as long as in the 
same class of medication, will not be considered a deviation of this inclusion criterion. Subjects may be on chronic or as needed 
(prn) medications if, in the opinion of the site PI or appropriate sub-investigator, they pose no additional risk to subject safety 
or assessment of reactogenicity and immunogenicity and do not indicate a worsening or treatment of continued symptoms of 
medical diagnosis or condition. Note: Low dose topical, corticosteroids as outlined in the Subject Exclusion Criteria (see 
Section 5.2) as well as herbals, vitamins and supplements are permitted. 
6. Oral temperature is less than 100.0℉. 
7. Pulse is 47 to 100 beats per minute, inclusive. 
8. Systolic blood pressure is 85 to 140 mmHg, inclusive. 
9. Diastolic blood pressure is 55 to 90 mmHg, inclusive. 
10. Screening laboratories (WBC, Hgb, PLTs, Sodium, Potassium, Bicarbonate, Calcium, Cr, non-fasting 
glucose, potassium, ALT, AST, TBIL, and urine protein and glucose) are within acceptable parameters2. 
2Hematology, blood chemistry and liver enzymes must be Grade 1 or less at screening (Refer to Table 11, Table 12, and Table 
13, respectively); urine glucose negative and urine protein no greater than trace at screening for subjects to qualify for 
randomization and vaccination. 
11. Negative test for HIV, hepatitis B virus (HBV), and hepatitis C virus (HCV) at screening blood draw. 
12. Women of childbearing potential3 must use an acceptable contraception method4 from at least 30 days 
before the first study vaccination until 30 days after the second study vaccination. 
3Not sterilized via, bilateral oophorectomy, salpingectomy, hysterectomy, or successful Essure® placement (permanent, non-
surgical, non-hormonal sterilization) with documented radiological confirmation test at least 90 days after the procedure, and 
still menstruating or <1 year has passed since the last menses if menopausal. 
4Includes non-male sexual relationships, full abstinence from sexual intercourse with a male partner, monogamous relationship 
with vasectomized partner who has been vasectomized for 180 days or more and shown to be azoospermic prior to the subject 

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receiving the study vaccination, effective intrauterine devices, NuvaRing®, tubal ligation, and licensed hormonal methods such 
as implants, injectables or oral contraceptives (i.e. “the pill”). 
13. Women of childbearing potential must have a negative serum pregnancy test at screening and a negative 
urine pregnancy test within 24 hours prior to each study vaccination. 
14. Sexually active males must agree to use a medically acceptable form of contraception5 in order to be in 
this study and must agree to continue such use until day 30 after the last vaccination.  
5Medically acceptable contraceptives include: (1) surgical sterilization (such as a vasectomy), or (2) a condom used with a 
spermicide.  Contraceptive measures such as Plan B™, sold for emergency use after unprotected sex, are not acceptable 
methods for routine use. 
 
 
 

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5.2 
Exclusion Criteria 
1. Have an acute illness1 or acute febrile illness (oral temperature  ≥ 38℃ [100.4℉]), as determined by the 
site PI or appropriate sub-investigator, within 72 hours prior to study vaccination. 
1An acute illness which is nearly resolved with only minor residual symptoms remaining is allowable if, in the opinion of the 
site PI or appropriate sub-investigator, the residual symptoms will not interfere with the ability to assess safety parameters as 
required by the protocol. 
2. Have any medical disease or condition that, in the opinion of the site PI or appropriate sub-investigator, 
is a contraindication to study participation2. 
2Including acute, subacute, intermittent or chronic medical disease or condition that would place the subject at an unacceptable 
risk of injury, render the subject unable to meet the requirements of the protocol, or may interfere with the evaluation of 
responses or the subject’s successful completion of this trial. 
3. Have immunosuppression as a result of an underlying illness or treatment, a recent history or current use 
of immunosuppressive or immunomodulating disease therapy. 
4. Use of anticancer chemotherapy or radiation therapy (cytotoxic) within 3 years prior to study vaccination. 
5. Have known active or recently active (12 months) neoplastic disease or a history of any hematologic 
malignancy. Non-melanoma, treated, skin cancers are permitted. 
6. Known allergy to components of the study product3. 
3Including the following: aluminum hydroxide, sorbitol, potassium chloride, sodium chloride and polysorbate80 (Tween80). 
7. Unstable seizure disorder (defined as requiring medication for seizure control or with seizure activity 
within the past 3 years). 
8. Have a history of alcohol or drug abuse within 5 years prior to study vaccination. 
9. Have any diagnosis, current or past, of schizophrenia, bipolar disease or other psychiatric diagnosis that 
may interfere4 with subject compliance or safety evaluations. 
4As determined by the site PI or appropriate sub-investigator. 
10. Have been hospitalized for psychiatric illness, history of suicide attempt, or confinement for danger to self 
or others within 5 years prior to study vaccination. 
11. Have a history of asthma, other than mild, well-controlled asthma5. 
5Cold or exercise induced asthma controlled with inhaled medications other than inhaled corticosteroids is permissible. 
Participants should be excluded if they require daily bronchodilator use, or have had an asthma exacerbation requiring 
oral/parenteral steroid use or have used theophylline or inhaled corticosteroids in the past year. 
12. Have a history of diabetes mellitus. 
13. Have taken oral or parenteral (including intra-articular) or chronic topical corticosteroids of any dose 
within 30 days prior to study vaccination6. 
6Corticosteroid nasal sprays for allergic rhinitis are permissible. Persons using a topical corticosteroid for a limited duration 
for mild uncomplicated dermatitis such as poison ivy or contact dermatitis may be enrolled the day after their therapy is 
completed. 
14. Have taken high-dose inhaled corticosteroids7 within 30 days prior to study vaccination. 
7High-dose defined as per age as using inhaled high-dose per reference chart in the National Heart, Lung and Blood Institute 
Guidelines for the Diagnosis and Management of Asthma (EPR-3) or other lists published in UPTODATE. 
15. Received or plan to receive a licensed, live vaccine within 30 days before or after each study vaccination. 

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16. Received or plan to receive a licensed, inactivated vaccine or allergy desensitization shot within 14 days 
before or after each study vaccination. 
17. Received an experimental agent8 within 30 days prior to the study vaccination or expect to receive another 
experimental agent9 during the trial-reporting period10. 
8Including vaccine, drug, biologic, device, blood product, or medication. 
9Other than from participation in this trial. 
10Approximately 7 months after the first study vaccination. 
18. Are participating or plan to participate in another clinical trial with an interventional agent11 that will be 
received during the trial-reporting period12. 
11Including licensed or unlicensed vaccine, drug, biologic, device, blood product, or medication. 
12Approximately 7 months after the first study vaccination. 
19. Female subjects who are breastfeeding or plan to breastfeed from the time of the first study vaccination 
through 30 days after the last study vaccination. 
20. Receipt of blood products or immunoglobulin within six months prior to enrollment. 
21. Donation of a unit of blood within 60 days prior to enrollment or intends to donate blood during the study 
period. 
22. Body mass index (BMI) ≥ 35. 
23. History of a visit to South America, sub-Saharan Africa or Southeast Asia lasting one month or more. 
24. Planned travel to areas known to be endemic with Yellow Fever virus during the study period. 
25. History of military service. 
26. History of vaccination against dengue, yellow fever, tick-borne encephalitis, or Japanese encephalitis. 
27. History of vaccination with other flavivirus candidate vaccines. 
28. Attended primary (grade) school in Austria, Germany, Japan, South Korea, India, Thailand, Nepal, 
Vietnam, or Taiwan (locations where the tick-borne encephalitis vaccine is given). 
29. History of yellow fever. 
30. Plan to have a major change in exercise routine or perform strenuous exercise from 72 hours before any 
dose of study vaccine/placebo and for 72 hours before any safety laboratories13. 
13Safety laboratories are obtained on Days 4 and 15 following each dose of study product. 
31. Contraindication to intramuscular vaccination of either upper arm (for example, due to lymphadenectomy 
or obscuring tattoos). 
 
5.3 
Exclusion of Specific Populations 
Children and pregnant women are excluded from this first-in-human Phase 1 study because insufficient 
information is available on the safety of HydroVax-002 YFV to judge potential risks for children and pregnant 
women. 
 

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5.4 
Inclusion of Vulnerable Participants 
Not applicable. 
 
5.5 
Lifestyle Considerations 
During this study, participants are asked to: 
• 
Avoid getting pregnant during the study from the time of screening until 30 days after the second vaccination 
• 
Use an acceptable contraception method, if a woman of childbearing potential, from 30 days before the first 
study vaccination until 30 days after the second study vaccination 
• 
Use a medically acceptable form of contraception, if a sexually active male,  until day 30 after the last 
vaccination 
• 
Refrain from receiving another experimental agent or participating in another clinical trial with an 
interventional agent 
• 
Refrain from donating blood during the study period 
• 
Refrain from a major change in exercise routine or strenuous exercise from 72 hours before any dose of study 
vaccine/placebo  and for 72 hours before each blood collection for clinical laboratory tests (Vaccination Days 
and Days 4 and 15 following each dose) 
• 
Abstain from travel to areas known to be endemic with Yellow Fever virus until 6 months after vaccination 
 
5.6 
Screen Failures 
Following consent, after the screening evaluations have been completed, the investigator or designee is to review 
the inclusion/exclusion criteria and determine the subject’s eligibility for the study.  If there is any uncertainty, 
the PI should make the decision on whether a potential subject is eligible for enrollment. 
 
Only the following information will be collected on screen failures: demographics (age, screen number, sex, 
ethnicity, and race) and reason for ineligibility. Subjects who are found to be ineligible will be told the reason for 
ineligibility. 
 
Individuals who do not meet the criteria for participation in this study (screen failure) because of a laboratory 
value that is outside the range of eligibility, but is thought to be due to an acute condition or due to laboratory 
error, may be repeated once  
 
5.7 
Strategies for Recruitment and Retention 
 
Recruitment 
Potential participants will be selected from existing participant registries, through advertising or by word of mouth 
in the Duke University, Durham, NC area and surrounding communities. Recruitment staff will contact interested 
individuals to assess their eligibility for this study. It is anticipated that the demographic composition of subjects 
for this study will be representative of the respective region. 
 

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The IRB will approve the recruitment process and all materials provided prior to any recruitment to prospective 
subjects directly. 
 
Screening will begin with a brief discussion with study staff. Some people will be excluded based on demographic 
data and medical history (i.e., pregnant, prior travel, prior vaccination, etc.). Information about the study will be 
presented to potential subjects and questions will be asked to determine potential eligibility. Screening procedures 
can begin only after informed consent is obtained. 
 
 
Retention 
Retention of subjects in this trial is especially important for determining the primary and secondary endpoints. As 
such, after vaccination, participating subjects will be reminded of subsequent study visits and every effort will be 
made to accommodate the subject’s schedule to facilitate follow-up within the specified visit window. 
Additionally, there are many circumstances that influence the ability to obtain outcome information after 
vaccination.  Follow-up visits may be conducted by phone if in person visits are not feasible. Lastly, compensation 
strategies will be implemented to encourage subjects to return for follow-up assessments and sample collections.  
 
 
Compensation Plan for Subjects 
Compensation will be determined locally at the study site and in accordance with local IRB regulations and 
approval. 
 
 
Costs 
There is no cost to subjects for the research tests, procedures/evaluations, and study product while taking part in 
this trial.  Procedures and treatment for clinical care may be billed to the subject, subject’s insurance or third 
party.   
 
 
 

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 STUDY PRODUCT 
6.1 
Study Product and Administration 
In this dose escalation study, the first 13 subjects will be randomized (10:3) to receive two doses of either 1 mcg 
HydroVax-002 YFV IM injection or placebo IM injection 28 days apart.  The second 12 subjects will be 
randomized (5:1) to receive two doses of 5 mcg HydroVax-002 YFV IM injection or placebo IM injection 28 
days apart. 
 
 
Study Product Description 
6.1.1.1 
Vaccine 
The HydroVax-002 YFV vaccine is a yellow fever virus, Vero cell tissue culture-derived vaccine inactivated with 
hydrogen peroxide, formulated with 0.1% aluminum hydroxide. HydroVax-002 YFV drug product contains 5 
mcg purified whole virus YFV formulated in a volume of 0.5 mL/dose with 0.1% aluminum hydroxide and 10% 
sorbitol, 0.001% polysorbate 80 in phosphate-buffer [pH = 7.5] with 350 mM sodium chloride. A single 0.5 mL 
dose of HydroVax-002 YFV may also contain residual amounts of Vero host cell DNA (<1 ng/0.5 mL) or Vero 
host cell protein (<300 ng/0.5).  HydroVax-002 YFV does not contain antibiotics or preservatives.  The stoppers 
used for single-dose vials do not contain latex. 
 
6.1.1.2 
Placebo and Diluent 
Sodium Chloride Injection USP, 0.9% (NaCl 0.9%, Normal Saline) will be used as the placebo and diluent. 
 
 
Dosing and Administration 
Based on dosing group assignment, subjects will receive either 1 mcg, 5 mcg, or placebo per vaccination dose 
administered in the non-dominant arm as a 0.5 mL intramuscular (IM) injection on days 1 and 29.  Following 
vaccination, subjects will be observed for a minimum of 30 minutes.  
 
 
Dose Escalation 
For this Phase 1 first-in-human trial, a conservative dose escalation design will be used, in which each dose group 
of 10 vaccine recipients, and a total of 5 placebo recipients, is split into two subgroups. The first subgroup, termed 
the sentinel subgroup, includes 3 vaccine and 1 placebo recipient. In each of the two (1 mcg and 5 mcg) dose 
phases, enrollment is halted after vaccination of the sentinel subgroup. This allows the opportunity to identify 
potential early vaccine related adverse events in this subgroup prior to exposure of additional subjects, a design 
which decreases the risk of multiple serious vaccine related adverse events compared to sequential vaccination of 
the entire dose group without halting [34]. After enrollment of the expanded subgroups, escalation to the next 
dose group will be delayed until 14 days of safety data following the second vaccination (corresponding to study 
Day 15) is available from all subjects in the preceding dose group. 
 
 
Dose Modifications 
Not applicable. 
 

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Overdosage 
Overdosage is not anticipated in the context of a clinical trial.   Subjects will be given intramuscular injections on 
Days 1 and 29.  No study product will be administered outside the clinical setting.  
 
6.2 
6.2 Preparation/Handling/Storage/Accountability 
 
Acquisition and Accountability 
 
 
 
 
 
 
 
 
  
 
 
Sodium Chloride Injection USP 0.9% for placebo and diluent will be sourced by the Duke Investigational Drug 
Service. 
 
After receipt of the study product, the principal investigator is responsible for distribution and disposition of these 
study products and has ultimate responsibility for drug accountability. The principal investigator may delegate to 
the research pharmacist responsibility for study vaccine accountability.  The research pharmacist must maintain 
and document logs of receipt, accountability, vaccine dilution and dispensation, storage conditions, and disposal.  
These study product accountability and dispensing logs must be maintained in the study file.  
 
Upon completion of the study and the final monitoring visit, used and unused vials of HydroVax-002 YFV and 
Placebo/diluent (sterile 0.9% NaCl) will be retained until monitored and released for disposition. For detailed 
information regarding final disposition of study vaccine and placebo/diluent see the protocol-specific MOP. 
 
 
Formulation, Appearance, Packaging, and Labeling 
HydroVax-002 YFV vaccine will be supplied to the investigative site in crystal zenith 2 mL vials containing 5 
mcg of purified, inactivated whole YFV virus formulated with alum per 0.5 mL with stopper, crimp seals, and 
flip-off cap. Each vial will contain a fill volume of 0.6 mL. The study vaccine is a white, opaque, non-uniform 
suspension that becomes homogeneous upon shaking.  
 
Sodium Chloride Injection USP 0.9% is a sterile, nonpyrogenic, isotonic solution of sodium chloride and WFI.  
It will be used to dilute the vaccine and will be used as the placebo.  
 
Further details regarding formulation, packaging and labeling are included in the protocol-specific Manual of 
Procedures (MOP) and the Investigator’s Brochure for the HydroVax-002 YFV vaccine. 

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Product Storage and Stability 
Study vaccine and the normal saline placebo/diluent will be shipped and stored refrigerated at 2℃ to 8℃. Study 
vaccine and the placebo/diluent should not be frozen.  
 
During storage, HydroVax-002 YFV vials should be kept in a lightproof container. During storage, a clear liquid 
with a white precipitate will be observed; this is the alum adjuvant and this appearance is to be expected following 
refrigerated storage. 
 
 
Preparation 
Study vaccine preparation, including vaccine dilutions for the various dosing groups, will be performed by the 
site research pharmacist on the same day of study vaccine administration.  Detailed information regarding the 
handling of vaccine, including preparation of the dilutions, labeling, storage, and administration of study vaccine 
will be provided in the protocol-specific MOP. 
 
Aseptic technique will be used for the administration of each dose of study product using a sterile needle 
appropriate in length for each participant, as described in the protocol-specific MOP. 
 
6.3 
Measures to Minimize Bias: Randomization and Blinding 
 
Treatment Assignment Procedures 
Per International Conference on Harmonisation (ICH) guideline E6: Good Clinical Practice (GCP), screening 
records will be kept at the clinical site to document the reason why an individual was screened, but failed trial 
entry criteria. The reasons why individuals failed screening will be recorded in the Coordinating Center’s (CC) 
Advantage eClinical® (Electronic Data Capture System). 
 
Once consented and upon entry of demographic data and confirmation of eligibility for the trial, the subject will 
be assigned to a treatment arm within each dose group (low dose and high dose) subjects will be randomly 
assigned to receive the study test product or placebo. Subjects will be block randomized so that 1 of the first 4 
sentinel subjects will receive placebo and the other 3 active vaccine. In the expanded groups, 7 of the 8 or 9 
subjects will be randomized to vaccine and 1 or 2 to placebo. Subjects will receive the same treatment for both 
vaccinations. 
 
 
Randomization 
Enrollment of subjects will be done online using the enrollment module of Advantage eClinical®. The 
randomization code will be prepared by statisticians at the CC and included in the enrollment module for the trial. 
Advantage eClinical® will generate a treatment assignment for each subject after the demographic and eligibility 
data have been entered into the system. A designated individual at each site will be provided with a code list for 
emergency unblinding purposes, which will be kept in a secure place. 
 

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Instructions for use of the enrollment module are included in the Advantage eClinical® User’s Guide. Manual 
back-up procedures and instructions are provided for use in the event that the site temporarily loses access to the 
Internet or the online enrollment system is unavailable. 
 
 
Blinding and Masking Procedures 
This is a partially-blinded clinical trial. 
 
Study staff and investigators will not be blinded to dose group but, within dose groups, subjects, investigators, 
and study staff other than unblinded site research pharmacist (or other designated unblinded personnel 
administering study product) will be blinded as to the subject's treatment assignment (vaccine vs. placebo). 
Laboratory personnel performing antibody assays will be blinded to dose group and treatment assignment. 
 
The randomization scheme will be generated by the CC and provided to unblinded study personnel (i.e., research 
pharmacists performing study vaccination preparations and unblinded study vaccine administrators) at the clinical 
site. 
 
An unblinded site research pharmacist will prepare the study product and an unblinded research nurse will inject 
the study product per the randomization assignment. The pharmacist will conceal the contents of the syringe by 
wrapping the syringe barrel with an overlay, opaque tape, or other equivalent material. In addition, the subject 
will be asked to look away when the vaccine is being administered. 
 
The unblinded site research pharmacist and unblinded research nurse will not be involved in study-related 
assessments or have subject contact for data collection following study vaccine administration. 
 
6.3.3.1 
Unblinding 
A copy of subject treatment assignments will be retained at the site in a secure file by the unblinded staff. In the 
event of a medical emergency when knowledge of the treatment assignment will influence subject’s care, the 
Principal Investigator may be provided with the treatment assignment. The Principal Investigator must contact 
NTI and document the event with information regarding the reasons for unblinding. After discussion with NTI, 
the Principal Investigator may receive the treatment assignment from the data coordinating center or if necessary, 
from the on-site secure file by the unblinded staff. 
 
6.4 
Study Intervention Compliance 
Each dose of study product will be administered by a member of the clinical research team who is qualified and 
licensed to administer study product. Administration site, date, and time will be documented on the case report 
form (CRF) and entered into the eCRF. 
 
6.5 
Concomitant Therapy 
All concomitant medications taken within 60 days prior to signing the ICF will be reviewed with subjects to 
determine stability of chronic diseases and eligibility. Medications reported in the eCRF are limited to those taken 

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within 30 days prior to the first dose of study vaccine. Concomitant medications will be reviewed at every study 
visit through Visit 07 (29 days following receipt of the second dose of study product).  
 
Women of childbearing potential in a heterosexual relationship must agree to use true abstinence or use at least 
one acceptable primary form of contraception through 30 days following the second dose of study vaccine. 
 
Prohibited medications: 
• 
Medications that may be associated with impaired immune responsiveness including immunosuppressive or 
immunomodulating therapies. 
• 
Anticancer chemotherapy within 3 years prior to vaccination. 
• 
Medications used to control seizures within the past 3 years. 
• 
Daily bronchodilators, oral or parental steroids, theophylline, or inhaled corticosteroids to treat an asthma 
exacerbation within the past year. 
• 
Oral, parenteral, intra-articular, chronic topical steroids or high-dose inhaled steroids within 30 days. 
• 
Investigational drug, vaccine, biologic, blood product, or medication within 30 days prior to receipt of the 
first dose of study product or during the study period. 
• 
A live vaccine within 30 days of receipt of study product. 
• 
An inactivated vaccine or allergy desensitization shot within 14 days of receipt of study product. 
• 
Blood products or immunoglobulin during the six months prior to enrollment. 
 
 
Rescue Medicine 
Not applicable. 
 
 
 

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 Study Intervention Discontinuation and Participant 
Discontinuation/Withdrawal 
7.1 
Halting Criteria and Discontinuation of Study Intervention 
 
Study Halting Criteria 
Additional enrollment and study interventions/administration of study products in this trial will be halted for SMC 
review/recommendation if any of the following are reported: 
• 
Any subject experiences ulceration, abscess, or necrosis at the injection site related to study product 
administration. 
• 
Any subject presents with laryngospasm, bronchospasm, hypotension or anaphylaxis occurring within 24 
hours following product administration that is considered related to study product. 
• 
Two or more subjects experience generalized urticaria (defined as occurring at more than two body parts) 
within 72 hours after administration of study product that is considered related to study product. 
• 
Any subject experiences an SAE after administration of study product and prior to the subject’s last visit that 
is considered related to study product. 
• 
Two or more subjects who received at least one dose of study vaccine to date, cumulative to all study vaccine 
administrations, across all treatment arms, experience the same Grade 3 (unsolicited AE or safety laboratory), 
in the same HLT by MedDRA coding, considered related to study product. 
 
This trial will also be halted for SMC review/recommendation if, within 7 days after administration of either study 
vaccination, any of the following occurs: 
• 
Three or more subjects who received at least one dose of study vaccine to date, cumulative to all study vaccine 
administrations, across all treatment arms, experience the same severe (Grade 3) study vaccine-related 
injection site reaction. Erythema and induration (hardness)/edema (swelling) will also be measured in mm but 
size will not be used as halting criteria. 
• 
Three or more subjects who received at least one dose of study vaccine to date, cumulative to all study vaccine 
administrations, across all treatment arms, experience the same severe (Grade 3) study vaccine-related 
subjective systemic reaction, for which the severity (grade) is corroborated by study personnel. 
• 
Three or more subjects who received at least one dose of study vaccine to date, cumulative to all study vaccine 
administrations, across all treatment arms, experience the same severe (Grade 3) study vaccine-related 
quantitative systemic reaction. 
Grading scales for solicited injection site and systemic (subjective and quantitative) reactions are included in 
Section 8.5.1.1. 
 
Grading scales for clinical safety laboratory AEs are included Appendix A. 
 

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If any of the halting rules are met following any subject receipt of any study vaccination, then this trial will not 
continue with the remaining enrollments or study vaccinations without a review by and recommendation from 
the SMC to proceed. 
 
NTI retains the authority to suspend additional enrollment and study interventions/administration of study 
products during this trial, as applicable. 
 
NTI is empowered to stop enrollment and study vaccinations if AEs that meet the halting criteria are reported. 
 
 
Individual Halting Criteria 
Individual Halting Criteria must be reviewed by the site PI prior to administration of the second study vaccination 
for each subject. It is the responsibility of the site to ensure that any subject meeting the below criteria are 
discontinued from receiving study product; however, the Emmes BMM or safety monitor may be consulted as 
needed. 
The second study vaccination will not be administered to a subject if any of the following criteria are met, as 
determined by the site PI: 
• 
Serious adverse event associated with study vaccination. 
• 
Pregnancy in a female subject. 
• 
Type 1 hypersensitivity associated with study product. 
• 
Grade 3 systemic reaction associated with study product. 
• 
Grade 3 local reaction (with the exception of erythema/induration) associated with study product (e.g., pain). 
• 
Grade 2 or higher fever within the first 24 hours following the initial vaccination. 
• 
Medical condition for which continued participation, in the opinion of the site principal investigator or 
appropriate sub-investigator, would pose a risk to the subject or would be likely to confound interpretation of 
the results. 
• 
Presence of signs or symptoms that could confound or confuse assessment of study vaccine reactogenicity. 
For subjects with injection site or systemic signs or symptoms, or with an acute illness, including but not 
limited to an oral temperature greater than or equal to 100.4℉, the second study vaccination should be 
postponed/deferred until signs, symptoms, or acute illness have resolved and still within the acceptable 
protocol-specified window for that visit. If outside this window, the Blinded Medical Monitor must first 
approve the second study vaccination and the documentation of approval should be filed in the subject’s chart. 
• 
Any unresolved or continuing solicited or unsolicited Grade 3 adverse event after first vaccination. An 
unresolved or continuing Grade 1 or Grade 2 adverse event is permissible unless, in the opinion of the site 
principal investigator or appropriate subinvestigator, it would render study vaccination unsafe or interfere 
with the evaluation of responses. 
• 
Grade 2 or higher clinical safety laboratory result that does not decrease to Grade 1 or below prior to the 
second study vaccination. For a subject to receive the second study vaccination, the most recently evaluated 
clinical safety laboratory values obtained prior to the second study vaccination must be Grade 1 or less. The 

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second study vaccination should be scheduled to occur within the acceptable protocol-specified window for 
that visit. If outside this window, the Blinded Medical Monitor must first approve the second study vaccination 
and the documentation of approval should be filed in the subject’s chart. 
• 
Grade 3 adverse event that occurs without alternative etiology in the 7 days following the first study 
vaccination. 
• 
Severe or sustained reaction or disability related to the first study vaccination. 
• 
New information becomes available that makes further participation unsafe. 
 
7.2 
Participant Withdrawal from the Study and Replacement 
 
Participant Withdrawal 
Subjects may voluntarily withdraw their consent for trial participation at any time and for any reason, without 
penalty or loss of benefits to which they are otherwise entitled. 
 
The site PI or appropriate sub-investigator may also withdraw a subject from receiving the study vaccine for any 
reason. 
 
A subject may withdraw or be withdrawn from the study for the following reasons:  
• 
Medical disease or condition, or any new clinical finding for which continued participation, in the opinion of 
the site PI or appropriate sub-investigator, would compromise the safety of the subject, or would interfere 
with the subject’s successful completion of this trial, or would interfere with the evaluation of responses (for 
example, has baseline significant laboratory abnormalities). 
• 
The subject withdraws consent. 
• 
Subject no longer meets eligibility criteria (see Sections 5.1 and 5.2). Note: Medication changes in the 60 days 
prior to enrollment, as specified in Subject Inclusion Criterion (See Section 5.1, Point Number 5), are 
exclusionary for receipt of the first study vaccination only. 
• 
As deemed necessary by the site PI or appropriate sub-investigator for noncompliance or other reasons. 
• 
The subject is lost to follow-up. 
• 
The study is terminated. 
• 
Any reason that, in the opinion of the investigator, precludes the subject's participation in the study. 
• 
New information becomes available that makes further participation unsafe. 
 
 
Subject Replacement 
Subjects who withdraw, or are withdrawn or terminated from the study, or are lost to follow-up after signing the 
informed consent form, randomization, and receipt of study vaccine will not be replaced. Subjects who withdraw, 
or are withdrawn or terminated from the study, or are lost to follow-up after signing the informed consent form 
and randomization but before receipt of study vaccine may be replaced. 
 

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7.3 
Lost to Follow-Up 
A participant will be considered lost to follow-up if he or she fails to return for scheduled visits and is unable to 
be contacted by the study site staff. 
 
7.4 
Follow up for subjects that discontinued study intervention 
The primary reason for withdrawal from this trial will be recorded on the Study Status data collection form (DCF). 
Subjects will be encouraged to complete the Early Termination Visit. The Early Termination Visit procedures are 
listed in Section 1.2. 
 
Although subjects are free to withdraw at any time or may be withdrawn by the site PI or appropriate sub-
investigator at any time (see Section 7.2.1), those subjects who receive only one dose of study vaccine will be 
encouraged to remain in this trial for follow-up safety assessments (may be conducted by phone call rather than 
in person) continuing through approximately 6 months after their last study vaccination. These subjects will also 
be encouraged to provide a venous blood sample for serological assays after their last study vaccination. See the 
protocol-specific Manual of Procedures (MOP) for alternate follow-up requirements. 
 
Every attempt will be made to follow all AEs, including solicited injection site and systemic reactions, unsolicited 
non-serious AEs, SAEs, and NOCMCs ongoing at the time of early withdrawal through resolution as per 
applicable collection times defined for the specific type of AE. 
 
In the case of subjects who fail to appear for a follow-up safety assessment, extensive effort (i.e., three 
documented contact attempts via phone calls 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 subject’s study records. 
 
The site PI or appropriate sub-investigator will inform the subject that already collected data will be retained and 
analyzed even if the subject withdraws or is withdrawn from this study. 
 
 

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 STUDY ASSESSMENTS AND PROCEDURES 
8.1 
Screening and Immunogenicity Assessments 
 
Screening Procedures 
Screening procedures may be done up to 28 days prior to enrollment (Day-28 to Day -1). After informed consent 
the following assessments are performed to determine eligibility and obtain baseline data: 
1. Take a focused medical history, including the following information: 
• 
History of any chronic medical conditions related to inclusion and exclusion criteria. 
• 
History of any medication allergies including history of hypersensitivity to any components of the study 
product. 
• 
Concomitant medications taken within 60 days prior to signing the ICF will be reviewed with subjects to 
determine stability of chronic diseases and eligibility. 
• 
Ask if they are participating in another clinical trial or plan to enroll in another clinical trial during the study 
period. 
2. Women of childbearing potentials should be counseled to either practice abstinence or use at least one 
primary form of contraception from 30 days prior to receipt of study product until 30 days after receipt of 
the last dose of study vaccine. 
3. Sexually active men should be counseled to use contraception from the date of receipt of the first dose of 
study vaccine until one month after receipt of the second dose of study vaccine. 
4. Women of childbearing potential must have a negative serum HCG pregnancy test at screening and a 
negative urine HCG pregnancy test prior to each dose of study product. 
5. Physical exam (standard exam at screening and targeted exam at enrollment). 
6. Height and weight measurement (Calculation of Body Mass Index). 
7. Vital signs measurement (including HR, BP, oral temperature). 
8. Blood for laboratory evaluations. 
• 
Serology at screening: HIV, Hepatitis B surface antigen, Hepatitis C antibody. 
• 
Chemistry at screening: sodium, potassium, bicarbonate, calcium, creatinine, glucose [non-fasting], ALT, 
AST, and total bilirubin. 
• 
Hematology at screening: white blood cells (WBCs), hemoglobin, and platelet count. 
• 
Urine for glucose and protein. 
 
Clinical screening laboratory evaluations will be performed locally by the Duke University Health System 
Clinical Laboratories.  Urine pregnancy tests will be performed by trained research staff.  The volume of venous 
blood to be collected is presented in Table 3. 
 

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The overall eligibility of the subject to participate in the study will be assessed once all screening values are 
available. Study subjects who qualify will be randomized into the Advantage eClinical system and all others will 
be registered as screen failures.  Results of screening testing will be discussed with the subject. If a positive result 
for HIV, hepatitis C or hepatitis B occurs or there is a laboratory result deemed to be clinically significant by the 
PI, the subject will be referred for appropriate follow-up.   Laboratory values that are outside the range of 
eligibility but are thought to be due to an acute condition or due to laboratory error may be repeated once. 
 
 
Immunogenicity Assessments 
8.1.2.1 
Humoral Immunogenicity Tests 
Neutralizing YFV-specific antibody responses following vaccination will be assessed by PRNT50 assays 
performed on blood samples obtained at Baseline (Day 1) and Days 15 and 29 after the first vaccination and at 
Days 15, 29, 57, and 180 following the second vaccination. 
 
8.1.2.2 
Specimen Preparation, Handling, and Shipping 
Blood specimens for YFV neutralizing assays will be processed as specified in the MOP and shipped monthly to 
Fisher BioServices. They will then be sent to OHSU (Beaverton, OR) for testing.  
 
8.2 
Safety and Other Assessments 
Study procedures are specified in the SoA.  A study physician licensed to make medical diagnoses and listed on 
Form 1572 will be responsible for all trial-related medical decisions. 
 
 
Physical examination 
A targeted physical exam will be performed at baseline and at subsequent in-person visits as indicated by changes 
in medical history.   
 
 
Safety Clinical Laboratory Tests 
Safety labs will be conducted on blood samples obtained on Days 4 and 15 after each vaccination and on the day 
of, but prior to, administration of the first and second vaccination (day 29 after first vaccination).  Safety labs 
include hematology tests (hemoglobin, WBC, and platelet count) and blood chemistry tests (sodium, potassium, 
bicarbonate, calcium, creatinine, glucose [non-fasting], ALT, AST, and total bilirubin).  
 
Urine will be tested by dipstick to evaluate glucose, protein, blood, and leukocyte esterase on days 1, 15  and  29 
after first vaccination and day 15 after second vaccination.  
 
Safety labs will be tested locally by Duke University Health System Clinical Laboratories.   
For laboratory values outside the site’s reference ranges, the site PI will document clinical significance in the 
eCRF.  Any result that is deemed clinically significant but does not meet the protocol’s grading criteria (Appendix 
A) should either be repeated by the study site, or the subject referred for appropriate clinical follow-up. 
 

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For any result that meets the protocol’s grading criteria (whether or not it is deemed clinically significant), the PI 
will also assess relationship to study product, action taken with product, and whether the subject was discontinued 
due to the event.  Graded laboratory abnormalities should be followed until either a return to normal or are judged 
to be stable, as determined by the PI. 
 
In the absence of a diagnosis, any clinical safety laboratory test result that meets the definition of an SAE as 
determined by the investigator (e.g., life-threatening) must be reported as an SAE using the AE/SAE form set. 
 
 
Viremia Detection Tests 
Blood samples will be obtained on Day 4 after each vaccination for YFV viremia testing. Viremia testing will 
include plaque assay and/or RT-PCR.  Positive results may be followed by additional testing (e.g., blind 
passage/amplification).  Frozen serum specimens for viremia testing will be shipped to Fisher BioServices. They 
will then be sent to OHSU (Beaverton, OR) for testing as needed. 
 
 
Additional Sera Samples 
Additional sera samples may be shipped to Fisher BioServices for storage. 
 
8.3 
Procedures to be Followed in the Event of Abnormal Laboratory Test Values or Abnormal 
Clinical Findings 
If a physiologic parameter, (e.g., vital signs, or laboratory value) is outside of the protocol-specified range, then 
the measurement may be repeated once if, in the judgment of the investigator, the abnormality is the result of an 
acute, short-term, rapidly reversible condition (e.g., stress, anxiety, or “white coat syndrome”). A physiologic 
parameter may also be repeated if there is a technical problem with the measurement caused by malfunctioning, 
or an inappropriate measuring device (i.e., inappropriate-sized blood pressure cuff). 
 
8.4 
Unscheduled Visit 
If the investigator deems the reaction warrants further evaluation or intervention, the investigator will give further 
instructions on the proper course of action, including a return to the clinic for immediate evaluation at an 
unscheduled visit if appropriate. 
 
8.5 
Adverse Events and Serious Adverse Events 
 
Definition of Adverse Events (AE) 
Adverse event means any untoward medical occurrence associated with the use of an intervention in humans, 
whether or not considered intervention-related (21 CFR 312.32 (a)). An AE can therefore be any unfavorable and 
unintended sign (including an abnormal laboratory finding), symptom, or disease temporally associated with the 
use of medicinal (investigational) product. 
 
Any medical condition that is present at the time that the subject is screened will be considered as baseline and 
not reported as an AE.  However, if the severity of any pre-existing medical condition increases, it should be 
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Adverse events can be further divided into solicited adverse events and unsolicited adverse events. Solicited 
adverse events are those for which the study team will specifically query the participant whether they occurred. 
Unsolicited adverse events are those events that the subject report occurring without being queried about the 
specific event.  
 
All AEs will be assessed for severity and relationship to study intervention (see Section 8.5.3.2). 
 
All AEs, solicited and unsolicited, will be captured on the appropriate data collection form.  Information to be 
collected for AEs includes event description, date of onset, assessment of severity, relationship to study product 
and alternate etiology (assessed only by those with the training and authority to make a diagnosis and listed on 
the FDA Form 1572 as an investigator), date of resolution, seriousness and outcome.  AEs occurring during the 
trial collection and reporting period will be documented appropriately regardless of relationship. In addition, all 
medically-attended adverse events (MAAEs) (i.e., events for which medical care from a healthcare provider is 
sought) will be collected through Study Visit 09. 
 
8.5.1.1 
Solicited Adverse Events/Reactogenicity 
Solicited adverse events are anticipated local and systemic adverse events for which consistent collection of 
information is desired. This may include: 
• 
Common and expected events according to the available knowledge about the product, or 
• 
Specific events of concern that should be ascertained on every participant. 
 
All subjects will be observed for at least 30 minutes after each vaccination to detect and treat any immediate 
adverse reactions. Subjects will be provided with a memory aid, digital, oral thermometer, and ruler for recording 
daily maximum oral temperature and systemic and local AEs beginning with the day of vaccination and 
continuing for the next 7 days. Subjects will be encouraged to take their temperature around the same time each 
day and also if they feel feverish, beginning with the day of vaccination. Subjects will be instructed on how to 
use the memory aid and how to rate any adverse events.  The subject will record temperature and the presence 
and intensity of post vaccination local reactogenicity events, including signs of redness and swelling and 
symptoms of pain and tenderness. The subjects will measure redness and swelling at the vaccination site with the 
measuring device provided. Systemic solicited symptoms will include feverishness, fatigue, headache, chills, 
nausea, new muscle pain (exclusive of the injection sites), aggravated muscle pain (increase of existing pain, 
exclusive of the injection site), new joint pain, and aggravated joint pain (increase of existing pain). The subjects 
will measure temperature with the thermometer provided.  Subjects will be instructed to notify the study center if 
they develop any severe reactions and/or fever equal to or exceeding 101.3°F. 
 
Subjects will also be asked to record any medications taken and any emergency room or physician visits (other 
than routine check-ups). The subject memory aid will be reviewed with the subject at subsequent clinic visits. 
 
After vaccination, antipyretics or analgesics should not be given routinely. Administration of an antipyretic or 
analgesic is to be recorded in the memory aid and entered into the CRF. 
 

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Definition of Serious Adverse Events (SAE) 
An adverse event or suspected adverse reaction is considered "serious" if, in the view of either the investigator or 
sponsor, it results in any of the following outcomes (21 CFR 312.32 (a)):  
• 
Death. 
• 
A life-threatening adverse event. 
• 
Inpatient hospitalization or prolongation of existing hospitalization. 
• 
A persistent or significant incapacity or substantial disruption of the ability to conduct normal life functions, 
or  
• 
A congenital anomaly/birth defect.  
 
Important medical events that may not result in death, be life-threatening, or require hospitalization may be 
considered serious when, based upon appropriate medical judgment, they may jeopardize the patient or subject 
and may require medical or surgical intervention to prevent one of the outcomes listed in this definition. Examples 
of such medical events include allergic bronchospasm requiring intensive treatment in an emergency room or at 
home, convulsions that do not result in inpatient hospitalization, etc. 
 
SAEs will be:  
• 
Assessed for severity and relationship to study product and alternate etiology (if not related to study product) 
by a licensed study physician listed on the FDA Form 1572 or by the Institution as the site Principal 
Investigator or Sub-Investigator.  
• 
Recorded on the appropriate SAE data collection form and eCRF.  
• 
Followed through resolution by a licensed study physician (for investigational new drug application [IND] 
studies, a physician listed on the FDA Form 1572 as the site Principal Investigator or Sub-Investigator). 
 
Reviewed and evaluated by the sponsor, the Safety Monitoring Committee (SMC) (periodic review unless 
related), and the IRB/IEC. 
 
8.5.2.1 
Suspected Unexpected Serious Adverse Reactions (SUSAR) 
A SUSAR is any SAE where a causal relationship with the study product is at least reasonably possible but is 
not listed in the Investigator Brochure, Package Insert, and/or Summary of Product Characteristics. 
 
 
Classification of an Adverse Events 
The determination of seriousness, severity, and causality will be made by an on-site investigator who is qualified 
(licensed) to diagnose, provide a medical evaluation of adverse events, and classify adverse events based upon 
medical judgment. This includes but is not limited to physicians, physician assistants, and nurse practitioners. 
 

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8.5.3.1 
Seriousness and Severity of Event 
Event seriousness will be determined according to the protocol definition of an SAE (Section 8.5.1).    All AEs 
or SAEs will be assessed for severity, according to the toxicity grading scales in Appendix A.  
 
For adverse events (AEs) not included in the protocol defined grading system, the following guidelines will be 
used to describe severity.  
• 
Mild (Grade 1):  Events that are usually transient and may require only minimal or no treatment or therapeutic 
intervention and generally do not interfere with the subject’s usual activities of daily living. 
• 
Moderate (Grade 2):  Events that are usually alleviated with additional specific therapeutic intervention.  The 
event interferes with usual activities of daily living, causing discomfort but poses no significant or permanent 
risk of harm to the research participant. 
• 
Severe (Grade 3):  Events interrupt usual activities of daily living, or significantly affects clinical status, or 
may require intensive therapeutic intervention.  Severe events are usually incapacitating. 
 
8.5.3.2 
Relationship to Study Intervention 
The licensed study physician’s assessment of an AE’s relationship to study product is part of the documentation 
process, but it is not a factor in determining what is or is not reported in this study. If there is any doubt as to 
whether a clinical observation is an AE, the event should be reported. The relationship to study product must be 
assessed for AEs using the terms: related or not related. In general, the study product must always be suspect. To 
help assess, the following guidelines are used:   
• 
Related – The AE is known to occur with the study intervention, there is a reasonable possibility that the study 
intervention caused the AE, or there is a temporal relationship between the study intervention and event. 
Reasonable possibility means that there is evidence to suggest a causal relationship between the study 
intervention and the AE. 
• 
Not Related – There is not a reasonable possibility that the administration of the study intervention caused the 
event, there is no temporal relationship between the study intervention and event onset, or an alternate etiology 
has been established. 
 
 
Time Period and Frequency for Event Assessment and Follow-up 
For this study all solicited AEs will be documented for the 8 days following study product administration. 
 
For this study, all unsolicited, non-serious AEs will be documented from Study Visit 01 (Day 1) through Study 
Visit 07 (Day 29 post second vaccination).  
 
All MAAEs, SAEs and new onset chronic medical conditions (NOCMC) will be documented from Study Visit 
01 (Day 1) through Study Visit 09 (final visit).  
 
Resolution of an AE is defined as the return to pre-treatment status or as stabilization of the condition, with the 
expectation that it will remain chronic.  Site PIs are required to follow all reported AEs until either resolution of 
the AE or until the event is stable, as determined by the site PI; all SAEs will be followed until either resolution 

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of the SAE or until the SAE is stable, as determined by the PI.  Subjects who have an unresolved SAE at either 
the final study visit or at discontinuation of study participation will continue to be followed, with the subject’s 
permission, until the SAE is stable/deemed chronic, as determined by the site PI.  
 
 
Adverse Event Reporting 
8.5.5.1 
Investigators Reporting of AEs 
Adverse Events, including local and systemic reactions not meeting the criteria for “serious adverse events,” will 
be captured on the appropriate case report form. Information to be collected for unsolicited adverse events 
includes event description, date of onset, investigator assessment of severity, relationship to study product, date 
of resolution of the event, seriousness, and outcome. 
 
 
Serious Adverse Event Reporting 
8.5.6.1 
Investigators Reporting of AEs 
Serious adverse events will be collected throughout the study until the final visit is completed. 
 
The following procedures will apply to all serious adverse events: 
• 
Assessed for severity and relationship to study product and alternate etiology (if not related to study product) 
by a licensed physician listed on the FDA Form 1572 as the principal investigator or subinvestigator. 
• 
Recorded on the appropriate serious adverse event report form and electronic case report form (eCRF). 
• 
Reviewed and evaluated by an Independent Safety Monitor, the SMC (periodic review unless related), NTI 
and the Institutional Review Board (IRB) (as indicated). 
• 
Reviewed and followed to resolution or stabilization by a licensed physician listed on the FDA Form 1572 as 
the principal investigator or subinvestigator. 
 
The Blinded Medical Monitor will review and assess the SAE for regulatory reporting and potential impact on 
study subject safety and protocol conduct. 
 
8.5.6.2 
Regulatory Reporting of AEs 
Following notification from the site Principal Investigator or appropriate sub-investigator, NTI, as the IND 
sponsor, will review the suspected unexpected serious adverse event (SUSAR) and ensure it is reported as an IND 
safety report to the FDA. Submissions of IND safety reports will be performed by Emmes on behalf of NTI. 
Emmes will notify all participating site Principal Investigators (i.e., all Principal Investigators to whom the 
sponsor is providing drug under its IND(s)) of potential serious risks from clinical studies or any other source, as 
soon as possible.  Emmes, on behalf of NTI, will report to the FDA any unexpected fatal or life-threatening 
suspected adverse reaction as soon as possible, but in no case later than 7 calendar days after the sponsor’s initial 
receipt of the information.  If the event is not fatal or life-threatening the IND safety report will be submitted 
within 15 calendar days after the sponsor determines that the information qualifies for reporting as specified in 
21 CFR Part 312.32.  Relevant follow up information to an IND safety report will be submitted as soon as the 
information is available.  Upon request from FDA, NTI (or Emmes, on NTI’s behalf) will submit to the FDA any 

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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.   
 
SAEs that are not SUSARs will be reported to the FDA at least annually in a summary format which includes all 
SAEs.   
 
 
Adverse Events of Special Interest 
This study will also assess New-Onset Chronic Medical Conditions (NOCMCs) from the day of receipt of the 
first dose of study product until study completion. NOCMCs are defined as any new ICD-10 diagnosis (10th 
revision of the International Statistical Classification of Diseases and Related Health Problems) that is applied to 
the subject during the course of the study, after receipt of the study agent, that is expected to continue for at least 
3 months and requires continued health care intervention.  NOCMCs will be assessed and recorded as unsolicited 
AEs.   
 
 
Reporting of Pregnancy 
Pregnancy is not an adverse event. Pregnancies occurring in study subjects will be reported via Advantage 
eClinical® on the Pregnancy Report form. No further study vaccinations will be administered to pregnant 
subjects, but if the subject has received a study vaccination and with the subject’s permission all study mandated 
blood samples will be obtained and the subject will continue in follow-up for safety events. Efforts will be made 
to follow all pregnancies reported during the course of the study to pregnancy outcome pending the subject’s 
permission. 
 
8.6 
Unanticipated Problems 
 
Definition of Unanticipated Problems (UP) 
The Department of Health and Human Services Office for Human Research Protections (OHRP) considers 
unanticipated problems involving risks to participants or others to include, in general, any incident, experience, 
or outcome that meets all of the following criteria: 
• 
Unexpected in terms of nature, severity, or frequency given (a) the research procedures that are described in 
the protocol-related documents, such as the Institutional Review Board (IRB)-approved research protocol and 
informed consent document; and (b) the characteristics of the participant population being studied; 
• 
Related or possibly related to participation in the research (“possibly related” means there is a reasonable 
possibility that the incident, experience, or outcome may have been caused by the procedures involved in the 
research); and 
• 
Suggests that the research places participants or others at a greater risk of harm (including physical, 
psychological, economic, or social harm) than was previously known or recognized. 
 
 
Unanticipated Problem Reporting 
The investigator will report unanticipated problems (UPs) to the reviewing Institutional Review Board (IRB) and 
to the Coordinating Center (CC). The UP report will include the following information: 

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• 
Protocol identifying information: protocol title and number, PI’s name, and the IRB project number. 
• 
A detailed description of the event, incident, experience, or outcome. 
• 
An explanation of the basis for determining that the event, incident, experience, or outcome represents an UP. 
• 
A description of any changes to the protocol or other corrective actions that have been taken or are proposed 
in response to the UP. 
 
To satisfy the requirement for prompt reporting, UPs will be reported using the following timeline:   
• 
UPs that are serious adverse events (SAEs) will be reported to the IRB and to the CC/study sponsor within 24 
hours of the investigator becoming aware of the event.  
• 
Any other UP will be reported to the IRB and to the CC/study sponsor within 5 days of the investigator 
becoming aware of the problem.  
 
 
Reporting Unanticipated Problems to Participants 
Not applicable. 
 
 
 

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Unsolicited AEs will be coded by the Medical Dictionary for Regulatory Activities (MedDRA) for preferred term 
and system organ class.  The rate and exact 95% confidence intervals of AEs in aggregate, and by MedDRA 
categories, will be computed.  SAEs will be reported by a detailed listing showing the type, MedDRA coding, 
relevant dates (vaccination and adverse event), severity, relatedness, and outcome for each event.  Primary 
outcome measures include: 
• 
Occurrence of all serious adverse events (SAEs) at any time during the study. 
• 
Occurrence of all Grade 3 unsolicited adverse events (AEs) from first vaccination through day 29 after the 
second vaccination. 
• 
Occurrence of all Grade 3 laboratory toxicities from first vaccination through day 15 after the second 
vaccination. 
• 
Occurrence of solicited local AE and reactogenicity signs and symptoms in the 7 days after each vaccination. 
• 
Occurrence of solicited systemic AE and reactogenicity signs and symptoms in the 7 days after each 
vaccination. 
• 
Occurrence of any AE through day 29 after the second vaccination. 
 
 
Analysis of Secondary and Exploratory Outcome Measures (Immunogenicity) 
Basic descriptive statistics, such as point estimates and 95% confidence intervals, and graphical summaries, such 
as the reverse cumulative distribution, of seroconversion rates and geometric mean titer (GMT) will be computed 
by intervention at baseline, 15 and 29 days after the first vaccination, and 15, 29, 57, and 180 days after the second 
vaccination. YFV-specific neutralizing antibody titers may be measured through a standard plaque reduction 
neutralizing titer [PRNT50] assay, or utilizing a Log10 Neutralizing Index (LNI) assay [5, 38].  For the purposes 
of this study, and in line with recent yellow fever vaccine clinical trials [7, 8], antibody titers will be assessed 
using the PRNT50 assay, and seroconversion will be defined as a PRNT50 neutralizing antibody titer of ≥1:10. The 
limit of detection in the neutralizing assay will be a titer of <10.  For the purposes of determining seroconversion 
rates and GMT, antibody titers of <10 will be assumed to be 5 (one dilution step below the assay limit value). 
 
Rates of seroconversion will be summarized by tabulating the frequency of positive responses by treatment group 
at Day 29 after first vaccination and Day 57 after second vaccination.  Response rates for each treatment group 
will be presented with their corresponding 95% confidence interval estimates at each time point. The relationship 
between the proportion of responders and dosage will be examined using logistic regression.  The distribution of 
antibody titers will also be graphically summarized using the reverse cumulative frequency distribution of titers 
for each treatment group.   
 
The PRNT50 titers at days 15 and 29 after first vaccination and at days 15, 29, and 57 following second vaccination 
will be summarized graphically for each treatment group.  The geometric mean titer (GMT) and pair-wise 
differences of each treatment group will also be presented with 95% confidence intervals.  The relationship 
between GMT and dosage will be examined using standard linear regression methods.  Secondary outcome 
measures include: 
• 
Percentage of subjects achieving seroconversion (1:10 in plaque reduction neutralizing titer [PRNT50] titer) 
at day 29 after first vaccination and at day 29 after second vaccination. 

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• 
Geometric mean neutralizing titers at days 15 and 29 after first vaccination and at days 15, 29, and 57 
following second vaccination. 
• 
Reverse cumulative distribution curve of neutralizing titers on Days 15 and 29 after first vaccination and at 
days 15, 29, and 57 after the second vaccination for each dose group and for all dose groups combined. 
 
Exploratory outcome measures include: 
• 
Geometric mean neutralizing titers at Day 180 following second vaccination. 
• 
Reverse cumulative distribution curve of neutralizing titers on Day 180 after the second vaccination for each 
dose group and for all dose groups combined. 
• 
Additional exploratory immunogenicity analyses utilizing the LNI assay may be conducted for comparison to 
historical datasets 
 
 
Timing of Analyses 
9.4.4.1 
Interim Safety Analyses 
The blinded medical monitor, clinical program manager, unblinded medical monitor and PI of the clinical 
laboratory will review data through the Day 15 visit after vaccination 1 for the 4 subjects in the sentinel groups.  
After the completion of Group 1 enrollment, the SMC will then review the data on all Group 1 subjects through 
the post 2nd vaccination Day 15 visit.  These reviews, however, may not involve any hypothesis testing and will 
not be considered in estimating the precision of any estimates made at the conclusion of the study. 
9.4.4.2 
 Final Analyses 
Clinical, safety, and reactogenicity data through approximately Day 57 after last study vaccination will represent 
the primary clinical database for the trial. Once all subjects have completed the Day 57 post dose 2 visit and the 
data are entered in the database, validated and monitored according to the clinical monitoring plan, the primary 
clinical database will be locked. Unblinded analyses of the primary and secondary safety, reactogenicity, and 
immunogenicity outcomes (Neutralization and viremia) and any supporting analyses of these outcomes described 
in the SAP will be performed by the SDCC and provided to the sponsor and PI.  Individual subject listings will 
not be generated at this point. The laboratory staff that run the assays for the Day 180 post dose 2 visit will remain 
blinded, and the assessment of relationship to study product of any SAEs that may be reported at the Day 180 
post dose 2 visit will be delegated to blinded sub-investigators at the clinical site.  While the results will not be 
used to make any decisions concerning the conduct of this trial, they may be used to make decisions on activities 
external to this trial such as the design of future trials of this vaccine.  Since this early analysis of the data is not 
intended to impact the conduct of the trial, it has no impact on Type I error and adjustments are not planned.  
 
The remaining analyses, including generation of all individual subject listings, will be completed and the CSR 
generated after the last subject’s last visit is completed, and the final clinical database, including all long-term 
safety follow-up data, is cleaned, monitored and locked.  
 
A formal statistical analysis plan (SAP) will be developed and finalized prior to unblinding for any analysis, 
which defines the analyses to be included in the CSR. 
 

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Sub-group Analyses 
Sub-group analysis (e.g, analysis based on age, sex, race/ethnicity or other demographic characteristics) is not 
planned due to the limited size of this Phase 1 clinical study. 
 
 
Tabulation of Individual participant Data 
Individual participant data will be listed by measure and time point after the last subject’s last visit is completed 
and the final clinical database, including all long-term safety follow-up data, is cleaned, monitored and locked. 
 
 

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 SUPPORTING 
DOCUMENTATION 
AND 
OPERATIONAL 
CONSIDERATIONS 
10.1 
Regulatory, Ethical and Study Oversight Considerations 
Each site principal investigator will obtain IRB approval for this protocol and any amendments to be conducted 
at his/her research site(s).   
 
NTI must receive the documentation that verifies IRB/IEC-approval for this protocol, informed consent, and 
associated documents prior to the recruitment, screening, and enrollment of subjects. 
 
 
Informed Consent Process 
Informed consent is a process that is initiated prior to an individual agreeing to participate in a trial and continuing 
throughout the individual’s trial participation. Before any study procedures are performed, informed consent will 
be obtained and documented. Subjects will receive a concise and focused presentation of key information about 
the trial, verbally and with a written ICF. The explanation will be organized and presented in lay terminology and 
language that facilitates understanding why one might or might not want to participate. The ICF must not include 
any exculpatory statements. 
 
The site PI or their designee will describe the protocol to potential subjects face-to-face. The key information 
about the purpose of the trial, the procedures and experimental aspects of the trial, risks and discomforts, any 
expected benefits to the subject, and alternative treatment will be presented first to the subject. 
 
Subjects will also receive an explanation that the trial involves research and a detailed summary of the proposed 
study procedures and study interventions/study products. This will include aspects of the trial that are 
experimental, the probability for random assignment to treatment arms, 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, alternative treatment/procedures that may be available, and the important 
potential benefits and risks of these available alternative treatment/procedures. 
 
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. 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 site PI) 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. The subjects will be informed that participation is voluntary and that they are free 
to withdraw from the study for any reason at any time without penalty or loss of benefits to which the subject is 
otherwise entitled. 

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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 monitors, auditors, IRB, 
NTI, and regulatory authorities will be granted direct access to the subject’s original medical records for 
verification of trial procedures and/or data without violating the confidentiality of the subject, to the extent 
permitted by the 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. Subjects will be informed whether private information 
collected from this research and/or samples/specimens will be used for additional research, even if identifiers are 
removed. 
 
Subjects will be allowed sufficient time to consider participation in the trial and have the opportunity to discuss 
the trial with their family, friends or legally authorized representative, or think about it prior to agreeing to 
participate. 
 
ICFs will be IRB-approved and subjects will be asked to read and review the ICF. Subjects must sign the ICF 
prior to starting any study procedures being done specifically for the trial. 
 
Once signed, a copy of the ICF will be given to the subjects for their records. The subject(s) may withdraw consent 
at any time throughout the course of the trial. The rights and welfare of the subjects will be protected by 
emphasizing to them that the quality of their medical care will not be adversely affected if they decline to 
participate in or withdraw from the trial. 
 
New information that significantly impacts the subject’s risk of receiving the study interventions/study products 
will be communicated by the site PIs or their designees to the subjects who consent to participate in the trial in 
accordance with IRB requirements. The ICF will be updated and subjects will be re-consented in accordance with 
IRB requirements, if necessary. Subjects will be given a copy of all ICFs that they sign. 
 
Study personnel may employ IRB-approved recruitment efforts prior to obtaining study consent if a patient-
specific screening consent is on record or if the IRB has agreed that chart review is allowed without a fully 
executed screening consent. In cases where there is not a patient-specific screening consent on record, Clinical 
Staff may pre-screen via chart review and refer potential subjects to the Research staff. Research staff would 
obtain written consent per the standard informed consent process before conducting protocol-specific screening 
activities. 
 
10.1.1.1 Requirements for Permission by Parents/Guardians and Assent by Children (in case of a 
minor)  
Not applicable. 
 

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10.1.1.2 
Other Informed Consent Procedures 
Not applicable. 
 
10.1.1.3 
Ethical Standard 
The PI will ensure that this trial is conducted in full conformity with principles of the Belmont Report: Ethical 
Principles and Guidelines for the Protection of Human Subjects of Research (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 21 CFR 56, as applicable. The site PIs will also ensure conformity with ICH E6 GCP and 
applicable federal regulations, guidance and guidelines for GCP and Clinical Trials with humans. 
 
10.1.1.4 
Institutional Review Board 
The institution engaged in this research will hold a current Federalwide Assurance (FWA) issued by the Office 
for Human Research Protections (OHRP) for federally funded research. The IRB must be registered with OHRP 
[OHRP-only or OHRP/FDA] as applicable to the research. The IRB FWA number will be provided to NTI. 
 
The site PI will obtain IRB approval for this protocol to be conducted at his/her research site(s) and send 
supporting documentation to NTI before initiating recruitment of subjects. The site PI will submit applicable 
information to the IRB on which it relies for the review, to conduct the review in accordance with 45 CFR 46, 
ICH E6 GCP guidelines, and as applicable, 21 CFR 56 (Institutional Review Boards), 21 CFR 50 (Protection of 
Human Subjects), and other federal, state and local regulations and guidance. NTI must receive the documentation 
that verifies IRB approval for this protocol, associated informed consent documents, and upon request, any 
recruitment material and handouts or surveys intended for the subjects, prior to the recruitment and enrollment of 
subjects. 
 
Any amendments to the protocol or consent materials will be approved by the IRB before they are implemented. 
IRB review and approval will occur at least annually throughout the enrollment and follow-up of subjects and 
may cease if annual review is no longer required by applicable regulations. The site PI will notify the IRB of 
protocol deviations and reportable SAEs in accordance with IRB requirements. 
 
 
Study Termination and Closure 
This study may be prematurely terminated if there is sufficient reasonable cause, including but not limited to: 
• 
Determination of unexpected, significant, or unacceptable risk to participants 
• 
Insufficient compliance to protocol requirements 
• 
Data that are not sufficiently complete and/or evaluable 
• 
Regulatory authorities 
 
If the study is prematurely terminated, the Principal Investigator (PI) will promptly inform study participants and 
the Institutional Review Board (IRB) and regulatory authorities as applicable. Study participants will be 
contacted, as applicable, and be informed of changes to study visit schedule. The PI will assure appropriate follow-
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The sponsor will notify regulatory authorities as applicable.   
 
 
Confidentiality and Privacy 
Subject confidentiality is strictly held in trust by the site PIs, other study personnel, the sponsor, and their agents. 
This confidentiality includes documentation, investigation data, subject’s clinical information, and all other 
information generated during participation in this trial. No information concerning this trial, or the data generated 
from this trial, will be released to any unauthorized third party without prior written approval of the subject and 
NTI. 
 
Subject confidentiality will be maintained when trial results are published or discussed in conferences and is 
extended to cover testing of samples/specimens. The study monitor or other authorized representatives of NTI as 
well as governmental regulatory agencies, such as the FDA, may inspect all documents and records required to 
be maintained by the site PI. This includes, but is not limited to, medical records (office, clinic or hospital) and 
pharmacy records for the subjects in this trial. The participating site will permit access to such records. 
 
All records will be kept locked and all computer entry and networking programs will be carried out with coded 
numbers only and with password-protected systems. All non-clinical samples/specimens, evaluation forms, 
reports, and other records that leave the site will be identified only by a coded number and will not be identified 
by the subject’s name. 
 
As this research is funded by the NIH, it is covered by NIH policy which effectively issues the research a 
Certificate of Confidentiality.  By this policy, researchers cannot be forced to disclose or provide, in any Federal, 
State, or local civil, criminal, administrative, legislative, or other proceeding, the name of such individual or any 
such information, document, or biospecimen that contains identifiable, sensitive information about the individual 
and that was created or compiled for purposes of the research, unless such disclosure or use is made with the 
consent of the individual to whom the information, document, or biospecimen pertains. 
 
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 

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research that the release is in compliance with applicable Federal regulations governing the protection of human 
subjects in research.  
 
 
Future Use of Stored Specimens for Secondary Research 
Subjects will be asked to consent to use of any remaining specimens for possible use in future secondary research 
studies as a condition of their study participation. A subject’s decision can be changed at any time prior to the end 
of the study by notifying the study doctors or nurses in writing. However, if a subject consents to future use and 
some of their blood has already been used for research purposes, the information from that research may still be 
used.  
 
Some samples may be stored at the local site and some at a central clinical storage facility. Samples may be shared 
with other investigators at other institutions. The samples will not be sold or used directly for production of any 
commercial product. No genetic testing will be performed on stored specimens. 
 
Each sample will be encoded (labeled) only with a barcode and a unique tracking number to protect subject’s 
confidentiality. There are no benefits to subjects in the collection, storage and subsequent research use of 
specimens. Reports about future research done with subject’s samples will not be kept in their health records. 
 
 
Key Roles and Study Governance 
Table 9. Table of Key Roles 
Principal Investigator 
 
 
 
 
 
 
 
 
Blinded Medical Monitor 
 
 
 
 
  
 
 
Unblinded Medical 
Monitor 
 
 
 
 
 
 

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Scientific Lead 
 
 
 
 
 
Clinical Program Manager 
 
 
 
 
 
 
Coordinating Center 
 
 
 
 
 
 
 
 
 
Safety Oversight 
10.1.6.1 
Safety Monitoring Committee (SMC) 
The study oversight will be conducted by a SMC which is an independent group of experts that will be established 
by NTI to monitor subject safety and advise NTI.  SMC members will be separate and independent of study 
personnel participating in this study and should not have scientific, financial or other conflicts of interest related 
to the study.  The SMC will consist of members with appropriate expertise to contribute to the interpretation of 
the data from this trial.   
 
The primary responsibilities of the SMC are to: 
 
• 
Review and evaluate the accumulated study safety data in a blinded fashion for participant safety after the 
low dose cohort prior to proceeding to the high dose cohort, study conduct and progress, and  
• 
Make recommendations to the sponsors concerning the continuation, modification, or termination of the trial.  
 
The operating rules of the SMC will be established in conjunction with NTI guidelines.  
 

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The SMC will review the safety laboratory results, reactogenicity data, and adverse event information collected 
through day 15 following Dose 2 for the Low Dose Cohort. SMC approval is required prior to initiation of 
enrollment in the High Dose Cohort.  
 
The SMC may also be asked to review safety information on the low dose or high dose sentinel groups as 
documented in Section 10.1.6.2. 
 
NTI or the SMC may convene the SMC on an ad hoc basis according to protocol criteria or if there are immediate 
concerns regarding observations during the course of the study.  Any AE/SAE that triggers a study halting rule 
will be reviewed by the Emmes BMM within 24 hours of its being reported, and, after confirming that a study 
halting rule was met, Emmes will alert the SMC chair to the situation and will provide a written narrative of the 
safety event. In addition, Emmes will notify the site PI that a halting rule has been triggered, and, upon approval 
by the SMC chair, will instruct the study site to suspend both study enrollment and study interventions (i.e., 
administering vaccine to subjects) until NTI provides explicit directions for resuming the suspended study 
activities. Concurrent with notification to the study site, Emmes will inform NTI that a halting rule has been 
triggered and that study activities, excepting follow-up with subjects for safety monitoring, have been paused 
pending the SMC’s review of the inciting event. The SMC will conduct an expeditious review of all relevant 
information, and a written summary of the committee’s discussion, to include any abiding concerns for subject 
safety, as well as any recommendations by the SMC regarding study continuation, will be submitted by Emmes 
to NTI. Any decision to suspend study enrollment and/or study interventions, either temporarily or permanently, 
is exclusively NTI’s. 
 
The SMC will review SAEs on a regular basis and ad hoc during the study. 
 
10.1.6.2 
Internal Safety Review Committee (ISRC) 
An internal safety review committee (ISRC), consisting of the blinded medical monitor, clinical program manager 
and the PI may be convened to provide an option for a rapid review of safety data collected through the day 8 
visit following dose one administered to the sentinel cohort of the low dose group and the sentinel cohort of the 
high dose group.   If within the sentinel group during the period from first vaccination through the day 8 visit, no 
individual halting rule was met, and no more than one unique Grade 3 adverse event (i.e., no same adverse event 
in more than one subject) was observed then the study will continue with administration of the second vaccination 
to the sentinel group and initiation of enrollment in the expanded dose cohort. The ISRC may be convened to 
assist with this objective determination.  If the criteria above will not be met, or at the request of the PI and/or 
NTI, the SMC will be convened to review the sentinel safety data.  If the SMC is convened for any reason for 
review of the sentinel data, the ISRC will not be convened for the cohort and the SMC will determine whether 
administration of the second vaccination to the sentinel cohort and initiation of enrollment in the expanded dose 
cohort in order for additional study vaccinations is to be given.  
 
 
Clinical Monitoring/Site Monitoring Plan 
Site monitoring is conducted to ensure that the human subjects’ protections, study and laboratory procedures, 
study interventions/administration of study products, and data collection processes are of high quality and meet 
sponsor and ICH E6 GCP guidelines and applicable federal regulations, and that this trial is conducted in 

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accordance with the protocol, protocol-specific MOP and applicable sponsor SOPs. NTI or its designee will 
conduct site-monitoring visits as detailed in the clinical monitoring plan. NTI-designated clinical monitors will 
verify that this trial is conducted, and data are generated, documented (recorded) and reported in compliance with 
the protocol, ICH E6 GCP guidelines and applicable regulatory requirements. Clinical monitoring reports will be 
submitted to NTI. 
 
Site visits will be made at standard intervals as defined by NTI and may be made more frequently as directed by 
NTI. Monitoring visits will include, but are not limited to, review of regulatory files, accountability records, 
eCRFs, ICFs, medical and laboratory reports, and protocol and GCP compliance. Site monitors will have access 
to each participating clinical site, study personnel and all study documentation according to the NTI-approved 
site monitoring plan. Study monitors will meet with site PIs to discuss any problems and actions to be taken and 
document visit findings and discussions. 
 
 
Quality Control and Quality Assurance 
Following a written NTI-accepted site quality management plan, the site is responsible for conducting routine 
quality assurance (QA) and quality control (QC) activities to internally monitor study progress and protocol 
compliance. The site PI will provide direct access to all study-related sites, source data/DCFs and reports for the 
purpose of monitoring and auditing by the sponsor, and inspection by local and regulatory authorities. The site PI 
will ensure all study personnel are appropriately trained and applicable documentation is current and maintained 
on site. 
 
The CC will implement quality control procedures beginning with the data entry system and generate data quality 
control checks that will be run on the database.  Any missing data or data anomalies will be communicated to the 
site(s) for clarification and resolution. 
 
 
Data Handling and Record Keeping 
The site principal investigator is responsible to ensure the accuracy, completeness, legibility, and timeliness of 
the data reported. 
 
Data collection forms will be derived from the electronic case report form and provided by the CC to record and 
maintain data for each subject enrolled in the study.  All data collection forms should be completed in a neat, 
legible manner to ensure accurate interpretation of data.  Black or blue ink is required to ensure clarity of 
reproduced copies.  When making a change or correction, 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 reported in the eCRF should be consistent with all source documents or the discrepancies should be 
explained. 
 
The sponsor will provide guidance to investigators on making corrections to the data collection forms and eCRFs. 
 

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10.1.9.1 
Data Collection and Management Responsibilities 
Data for this study will include safety, laboratory (safety and immunologic) and outcome measures (e.g., 
reactogenicity and immunogenicity). 
 
Clinical data (including AE/SAEs, concomitant medications, physical assessments, and reactogenicity data) will 
be entered into the CC’s Advantage eClinical® web-based 21 CFR Part 11-compliant Internet Data Entry System 
(IDES). The data system includes password protection and internal quality checks, such as automatic range checks 
to identify data that appear inconsistent, incomplete, or inaccurate.  Clinical data will primarily be entered directly 
from the data collection forms. 
 
All data collection forms must be reviewed by the clinical team and data entry staff, who will ensure that they are 
accurate and complete.  Adverse events must be graded, assessed for severity and causality, and reviewed by the 
site principal investigator or designee.  Data collection is the responsibility of the clinical trial staff at the site 
under the supervision of the site principal investigator.  During the study, the investigator must maintain complete 
and accurate documentation for the study. 
 
The CC will be responsible for data management, quality review, analysis, and reporting of the study data. 
 
A final clinical study report will be prepared following the last subject visit and upon completion of assays related 
to the immunogenicity endpoints. 
 
10.1.9.2 
Study Records Retention 
Study records and reports, including, but not limited to, eCRFs, source documents, ICFs, and study drug 
disposition records shall be maintained for 2 years after a marketing application is approved for the drug for the 
indication for which it is being investigated; or, if no application is to be filed or if the application is not approved 
for the drug, until 2 years after the investigation is discontinued and the FDA has been notified. These documents 
will be retained for a longer period, however, if required by local regulations. ICFs for future research use will be 
maintained as long as the sample/specimen exists. 
 
No records will be destroyed without the written consent of the sponsor. It is the responsibility of the sponsor to 
inform the site PI when these documents no longer need to be retained. The participating site must contact NTI 
for authorization prior to the destruction of any study records. 
 
10.1.9.3 
Source Records 
The  site will maintain appropriate medical and research records for this clinical trial, in compliance with ICH E6 
GCP Section 4.9 and regulatory and institutional requirements for the protection of confidentiality of subjects. 
The site will permit the study monitor or other authorized representatives of NTI or its representative as well as 
governmental regulatory agencies, such as the FDA, to examine (and when required by applicable law, to copy) 
clinical trial records for the purposes of quality assurance reviews, audits, monitoring and evaluation of the study 
safety and progress. These representatives will be permitted access to all source data, which include, but are not 
limited to, hospital records, clinical and office charts, laboratory notes, memoranda, evaluation checklists, 
pharmacy dispensing records, recorded data from automated instruments, copies or transcriptions certified after 

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verification as being accurate and complete, microfiches, photographic negatives, microfilm or magnetic media, 
x-rays, and subject files, and records kept at the pharmacy, at the laboratories and medico-technical departments 
involved in this clinical trial. Source data are all information, original records of clinical findings, observations, 
or other activities in a clinical trial necessary for the reconstruction and evaluation of the clinical trial. 
 
Interview of subjects is sufficient for obtaining medical history. Solicitation of medical records from the subject’s 
primary care provider is not required. 
 
 
Protocol Deviations 
A protocol deviation is any noncompliance with the study protocol, GCP or protocol-specific MOP requirements. 
The noncompliance may be either on the part of the subject, the site PI or other study personnel. As a result of 
protocol deviations, corrective actions are to be developed by the site and implemented promptly. 
These practices are consistent with ICH E6 GCP guidelines: 
 
• 
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 and other study personnel to use continuous vigilance to identify and report 
protocol deviations within five working days of identification of the protocol deviation, or within five working 
days of the scheduled protocol-required activity. Protocol deviations must be promptly reported to NTI, via the 
CC’s Advantage eClinical® system. 
 
Protocol deviations, as defined above, must be addressed on the appropriate DCF. A completed copy of the 
Protocol Deviation DCF must be maintained in the regulatory file as well as in the subject’s chart. Protocol 
deviations must be sent to the IRB in accordance with IRB requirements. The site PI and other study personnel 
are responsible for knowing and adhering to IRB requirements. 
 
 
Publication and Data Sharing 
 
10.1.11.1 Human Data Sharing Plan 
This study will be conducted in accordance with the following publication and data sharing policies and 
regulations: 
• 
National Institutes of Health (NIH) Public Access Policy, which ensures that the public has access to the 
published results of NIH funded research. It requires scientists to submit final peer-reviewed journal 
manuscripts that arise from NIH funds to the digital archive PubMed Central upon acceptance for publication. 
 
10.1.11.2 Genomic Data Sharing Plan 
Not applicable. 
 

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10.1.11.3 Publication 
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 all 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 ClinicalTrials.gov. 
 
For this clinical trial, the responsible party is NTI which will register this trial and post results. 
 
The responsible party does not plan to request certification of delayed posting. 
 
Refer to: 
• 
Public Law 110-85, Section 801, Clinical Trial Databases 
• 
42CFR11 
• 
NIH NOT-OD-16-149 
 
 
Conflict of Interest Policy 
The independence of this study from any actual or perceived influence, such as by the pharmaceutical industry, 
is critical.  Therefore, any actual conflict of interest of persons who have a role in the design, conduct, analysis, 
publication, or any aspect of this trial will be disclosed and managed. Furthermore, persons who have a perceived 
conflict of interest will be required to have such conflicts managed in a way that is appropriate to their 
participation in the design and conduct of this trial.  NTI has established policies and procedures for all study 
group members to disclose all conflicts of interest and will establish a mechanism for the management of all 
reported dualities of interest. 
 

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10.2 
Additional Considerations 
 
Research Related Injuries 
Immediate necessary medical care is available at Duke University Medical Center in the event that a subject is 
injured as a result of participation in this research study. However, there is no commitment by Duke University, 
Duke University Health System, Inc., or  Duke physicians to provide monetary compensation or free medical care 
in the event of a study–related injury. In addition, no long term medical care or financial compensation for 
research-related injuries will be provided by Najít Technologies, Inc. or the Federal government. 
 
 

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• Section 5.1, Inclusion Criteria, Point 10 – 
Removed BUN (blood urine nitrogen) from the 
list of screening laboratories to harmonize with 
the Schedule of Assessments, Table 2. 
• Section 6.1.2, Dosing and Administration – 
Clarified that the non-dominant arm will be the 
injection site for all administrations. 
• Section 7.1.2, Individual Halting Criteria – 
Included a preface statement indicating that the 
site PI must review Individual Halting Criteria for 
each subject prior to the administration of the 
second study vaccination. 
• Section 8.1.1, Screening Procedures – Removed 
SpO2 from the screening procedure list to 
harmonize with the Schedule of Assessments, 
Table 2. 
• Section 8.1.1, Screening Procedures – Added 
sodium, potassium, bicarbonate and calcium to 
the list of enrollment chemistry screens to 
harmonize with the Schedule of Assessments, 
Table 2. 
• Section 8.2.2, Safety Clinical Laboratory Tests – 
Added sodium, potassium, bicarbonate and 
calcium to the list of safety chemistry screens to 
harmonize with the Schedule of Assessments, 
Table 2.   Also clarified procedure for safety 
laboratory retests. 
• Section 8.5.1.1, Solicited Adverse 
Events/Reactogenicity – Updated temperature for 
instructing subjects to notify the study center 
from 101.1°F to 101.3°F to harmonize with Table 
7, Table of Fever Grading, and updated 
Fahrenheit temperature from 101.1°F to 101.3°F 
in Table 7 to match associated Celsius reading of 
38.5°C.  Also  Also updated the grading scale 
language in Table 4, Table of injection site 
symptom grading. 
• Sections 8.5.1.1 and 8.5.4 – Clarified  process for 
following an AE that is still present at EOS visit. 
this protocol several 
inconsistencies were 
observed between the 
Schedule of 
Assessments (SoA) 
shown in Table 2, and 
the narrative description 
of screening and safety 
procedures in Section 8.  
The indicated 
subsections in Section 8 
were updated to 
harmonize with the SoA 
Table. 
 
In addition, 
inconsistencies between 
Celsius and Fahrenheit 
values were discovered 
in Table 7.  Fahrenheit 
values were updated to 
match their Celsius 
counterparts. 
 
Further review by 
Emmes led to additional 
safety monitoring 
suggestions as indicates. 

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• 8.5.6.2, Regulatory Reporting of AEs – Provided 
clarifying language. 
• Table 9, Key Roles – Updated the Medical 
Monitor to Blinded Medical Monitor (BMM) and 
Medical Officer to Unblinded Medical Monitor 
(UMM). Clarified the specific roles for each 
throughout the protocol. 
• 10.1.6.1, Safety Monitoring Committee –  
Updated language on halting rule notification 
process. 
5.0 
27May2022 
• Section 1.1.5, Inclusion Criteria, Point 10 – 
Updated urine screening to specify that trace 
protein is not exclusionary at screening. 
• Section 5.1, Inclusion Criteria, Point 10 – 
Updated urine screening to specify that trace 
protein is not exclusionary at screening. 
• Appendix A, Table 14. Urine dipstick toxicity 
table. Updated table to indicate that trace urine 
protein is not exclusionary at screening 
 
Trace protein on urine 
screening may be 
commonly found and 
not be indicative of any 
underlying pathology.  
Exclusion modified to 
exclude only those with 
1+ or greater amounts of 
protein on urine 
screening. 
6.0 
12Jan2023 
• Immunogenicity analysis of the day 180 serum 
sample (Visit 09) was converted to exploratory 
analysis.  Changes were made in Sections 1.2, 
3.0, 9.4.3, 9.4.4 and 9.4.6. 
This change streamlined 
the analysis and 
reporting timeline for 
the clinical study 
without impacting any 
planned safety 
assessments. 

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APPENDIX A 
Toxicity Tables 
 
ABBREVIATIONS: Abbreviations utilized in the Table:  
ULN = Upper Limit of Normal  
LLN = Lower Limit of Normal  
NOTE: For any of the lab values listed below if the Grade 1 value conflicts with the ULN or LLN value, the ULN 
or LLN value takes precedence and the event will not be considered a Grade 1 Adverse Event.