Conditions: Dengue
Keywords: Drug Therapy
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 900
Healthy volunteers: yes

Brief summary:
The main purpose of this study is to assess the immunogenicity and safety of the concomitant and sequential administration of yellow fever (YF) vaccine and tetravalent dengue vaccine (TDV) in healthy participants aged 18 to 60 years living in country non-endemic for both dengue and YF.

Detailed description:
The vaccine tested in this study is TDV also known as TAK-003 (DENVax). TDV with concomitant and sequential administration of yellow fever (YF-17D) vaccine will be tested to assess immunogenicity and safety in healthy adult participants in non-endemic area(s) for both dengue and YF.

The study will enroll 900 healthy participants. Participants will be randomized to 3 groups in 1:1:1 ratio and will be administered concomitantly and sequentially. The 3 groups are:

* Group 1: YF-17D vaccine + placebo concomitantly administered on Day 1, first dose of TDV administered on Day 90 and second dose of TDV administered on Day 180.
* Group 2: first dose of TDV + placebo concomitantly administered on Day 1, second dose of TDV administered on Day 90 and YF-17D vaccine administered on Day 180.
* Group 3: first dose of TDV + YF-17D vaccine concomitantly administered on Day 1, second dose of TDV administered on Day 90 and placebo administered on Day 180.

This multi-center trial will be conducted in the United States. The overall time to participate in this study is 360 days. Participants will make multiple visits to the clinic with a 6-month follow up including a final visit at Day 360 for a follow-up assessment.

Arms:
arm 1: YF-17D vaccine, 0.5 mL injection, subcutaneously (SC) plus YF 17D + TDV placebo-matching 0.5 mL, injection, SC on Day 1, followed by TDV, 0.5 mL, injection, SC on Day 90 (first dose), followed by TDV, 0.5 mL, injection, SC on Day 180 (second dose). arm 2: TDV, 0.5 mL, injection, SC plus TDV placebo-matching, 0.5 mL injection, SC on Day 1 (first dose), followed by TDV, 0.5 mL, injection, SC on Day 90 (second dose), followed by YF-17D vaccine, 0.5 mL, injection, SC on Day 180. arm 3: TDV, 0.5 mL, injection, SC plus YF-17D vaccine, 0.5 mL, injection, SC on Day 1 (first dose), followed by TDV, 0.5 mL, injection, SC on day 90 (second dose), followed by TDV + YF 17D placebo-matching, 0.5 mL, injection, SC on Day 180.

Interventions: intervention 1: YF-17D intervention 2: TDV intervention 3: Placebo
Intervention descriptions: intervention 1: YF-17D SC injection. intervention 2: TDV SC injection. intervention 3: Normal Saline (0.9% NaCl) SC injection.

Protocol document:
Prot_000.pdf: 



Title: A Randomized, Observer-Blind, Placebo-Controlled, Phase 3 Trial to Investigate the 
Immunogenicity and Safety of a Tetravalent Dengue Vaccine Candidate and a Yellow Fever YF-17D 
Vaccine Administered Concomitantly and Sequentially in Healthy Subjects 
Aged 18 to 60 years in Non-Endemic Country(ies) 
NCT Number: NCT03342898
Protocol Approve Date: 28 November 2018
Certain information within this protocol has been redacted (ie, specific content is masked irreversibly from view 
with a black/blue bar) to protect either personally identifiable information (PPD) or company confidential 
information (CCI). 
This may include, but is not limited to, redaction of the following: 
Named persons or organizations associated with the study.
Proprietary information, such as scales or coding systems, which are considered confidential 
information under prior agreements with license holder.
Other information as needed to protect confidentiality of Takeda or partners, personal information, or 
to otherwise protect the integrity of the clinical study.


TITLE PAGE 
PROTOCOL 
A Randomized, Observer-Blind, Placebo-Controlled, Phase 3 Trial to Investigate the 
Immunogenicity and Safety of a Tetravalent Dengue Vaccine Candidate and a Yellow Fever 
YF-17D Vaccine Administered Concomitantly and Sequentially in Healthy Subjects 
Aged 18 to 60 years in Non-Endemic Country(ies) 
Immunogenicity and Safety of TDV Administered with a Yellow Fever Vaccine in Adults 
Sponsor: 
Takeda Vaccines, Inc. 
40 Landsdowne Street, 
Cambridge, MA 02139, 
USA 
Trial Identifier: 
DEN-305 
IND Number: 
014292 
EudraCT Number: 
Not Applicable 
Vaccine Name: 

Investigational vaccine: Takeda’s tetravalent dengue vaccine
candidate (TDV) comprised of a molecularly characterized,
attenuated dengue serotype 2 strain (TDV-2), a dengue serotypes 2/1
chimeric strain (TDV-1), a dengue serotypes 2/3 chimeric strain
(TDV-3), and a dengue serotypes 2/4 chimeric strain (TDV-4)

Licensed live yellow fever (YF-17D) vaccine

Placebo: saline solution
Date: 
28 November 2018 
Version: 
Version 4.0 
Property of Takeda: For non-commercial use only and subject to the applicable Terms of Use


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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 4 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
INVESTIGATOR AGREEMENT 
I confirm that I have read and that I understand this protocol, the Investigator’s Brochure, and any 
other product information provided by the Sponsor. I agree to conduct this trial in accordance with 
the requirements of this protocol and also protect the rights, safety, privacy, and well-being of trial 
subjects in accordance with the following: 
 The ethical principles that have their origin in the Declaration of Helsinki. 
 International Council for Harmonization of Technical Requirements for Pharmaceuticals for 
Human Use, E6 Good Clinical Practice: Consolidated Guideline. 
 All applicable laws and regulations, including, but not limited to those related to data privacy 
and clinical trial disclosure. 
 Regulatory requirements for reporting serious adverse events defined in Section 10.4 of this 
protocol. 
 Terms outlined in the Clinical Trial Site Agreement. 
 Appendix A – Responsibilities of the Investigator. 
I further authorize that my personal information may be processed and transferred in accordance 
with the uses contemplated in Appendix B of this protocol. 
 
Signature of Investigator 
Date 
Investigator Name (print or type) 
Investigator’s Title 
Location of Facility (City, State) 
Location of Facility (Country) 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 5 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
1.3 
Protocol Amendment Summary of Changes 
This document describes the changes in reference to the Protocol incorporating Amendment No. 2. 
The primary purpose of this amendment is to remove the planned Day 120 (Month 4 [M4]) interim 
analysis. Minor grammatical and editorial changes are included for clarification purposes only. 
Full details on changes of text are given in Section 1.3.2. 
1.3.1 Amendment History 
Date 
Amendment Number 
Amendment Type 
Region 
30 May 2017 
Initial Protocol 
Not applicable 
Global 
22 January 2018 
1 
Substantial 
Global 
28 November 2018 
2 
Non-substantial 
Global 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 6 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
1.3.2 Summary of Changes 
Amendment to Protocol Version 3.0, 22 January 2018 
Rationale for the Amendment: 
The rationale for removing the planned Day 120 (M4) interim analysis is that it is no longer 
required. 
The removal of this interim analysis has no impact on the final statistical analyses planned for 
this study, the scientific value of the study, the study conduct, or the safety of subjects enrolled. 
The administrative trial information has been updated and references list has been checked and 
updated as needed. 
Details of the changes are outlined below. In this section only (ie, not in the protocol body) all 
new text is shown in bold italics and any redundant text is marked using strikethrough. 
Property of Takeda: For non-commercial use only and subject to the applicable Terms of Use


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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 8 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
Section Description of change 
 
2.0; 
13.1.5 
An interim analysis on safety and immunogenicity data is planned when all subjects 
have completed Visit 4 (Day 120 [M4]). This analysis will be performed by a separate 
group of unblinded statisticians and programmers at a selected independent contract 
research organization (CRO) who will have access to individual vaccine assignment but 
will not be involved in subsequent trial conduct. The rest of the personnel involved in the 
conduct of the trial, including those at Takeda, the CRO, and the trial sites, will remain 
blinded to individual vaccine assignment until unblinding after trial completion 
(database lock for data through the Day 360 [Month 12] follow-up visit). No 
modification to the trial will be made based on the results of this interim analysis. More 
details regarding the interim analysis will be provided in the SAP. No interim analyses 
are planned. 
6.1 
Data collection will be by electronic Case Report Form (eCRF). Refer to Section 13.1.5 
for the planned interim analysis. 
 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 9 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
TABLE OF CONTENTS 
1.0 
ADMINISTRATIVE INFORMATION ..............................................................................2 
1.1 
Contacts..........................................................................................................................2 
1.2 
Approval ........................................................................................................................3 
1.3 
Protocol Amendment Summary of Changes..................................................................5 
1.3.1 Amendment History .................................................................................................5 
1.3.2 Summary of Changes ...............................................................................................6 
2.0 
TRIAL SUMMARY ..........................................................................................................13 
2.1 
Schedule of Trial Procedures .......................................................................................23 
3.0 
TRIAL REFERENCE INFORMATION ...........................................................................26 
3.1 
Trial-Related Responsibilities ......................................................................................26 
3.2 
Principal Investigator/Coordinating Investigator .........................................................26 
3.3 
List of Abbreviations ...................................................................................................27 
3.4 
Corporate Identification ...............................................................................................29 
4.0 
INTRODUCTION .............................................................................................................30 
4.1 
Background ..................................................................................................................30 
4.2 
Rationale for the Proposed Trial ..................................................................................31 
5.0 
TRIAL OBJECTIVES AND ENDPOINTS ......................................................................32 
5.1 
Objectives ....................................................................................................................32 
5.1.1 Primary Objective ..................................................................................................32 
5.1.2 Secondary Objectives.............................................................................................32 
5.2 
Endpoints .....................................................................................................................32 
5.2.1 Primary Endpoint ...................................................................................................32 
5.2.2 Secondary Endpoints .............................................................................................33 
6.0 
TRIAL DESIGN AND DESCRIPTION ...........................................................................34 
6.1 
Trial Design .................................................................................................................34 
6.2 
Justification for Trial Design, Dose, and Endpoints ....................................................36 
6.3 
Duration of Subject’s Expected Participation in the Entire Trial ................................36 
6.4 
Premature Termination or Suspension of Trial or Investigational Site .......................37 
6.4.1 Criteria for Premature Termination or Suspension of the Trial .............................37 
6.4.2 Criteria for Premature Termination or Suspension of Investigational Sites ..........37 
6.4.3 Procedures for Premature Termination or Suspension of the Trial or the 
Participation of Investigational Site(s)...................................................................37 
7.0 
SELECTION AND DISCONTINUATION/WITHDRAWAL OF SUBJECTS ...............38 
7.1 
Inclusion Criteria .........................................................................................................38 
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7.2 
Exclusion Criteria ........................................................................................................38 
7.3 
Criteria for Delay of Vaccination and/or Blood Sampling ..........................................40 
7.4 
Criteria for Early Trial Termination of a Subject ........................................................41 
7.5 
Criteria for Premature Discontinuation of Trial Vaccine Administration ...................42 
8.0 
CLINICAL TRIAL MATERIAL MANAGEMENT .........................................................44 
8.1 
Trial Vaccines and Materials .......................................................................................44 
8.1.1 Dosage Form, Packaging, and Labeling ................................................................44 
8.1.2 Storage ...................................................................................................................45 
8.1.3 Dose and Regimen .................................................................................................45 
8.2 
Trial Vaccine Assignment and Dispensing Procedures ...............................................46 
8.3 
Randomization Code Creation and Storage .................................................................46 
8.4 
Trial Vaccine Blind Maintenance ................................................................................47 
8.5 
Unblinding Procedure ..................................................................................................47 
8.6 
Accountability and Destruction of Sponsor-Supplied Vaccine(s) ...............................47 
9.0 
TRIAL PLAN ....................................................................................................................49 
9.1 
Trial Procedures ...........................................................................................................49 
9.1.1 Informed Consent Form .........................................................................................49 
9.1.2 Demographics, Medical History and Prior Medications ........................................49 
9.1.3 Documentation of Trial Entrance/Randomization .................................................50 
9.1.4 Physical Examination.............................................................................................50 
9.1.5 Vital Signs ..............................................................................................................51 
9.1.6 Immunogenicity Assessments ................................................................................51 
9.1.7 Safety Assessments ................................................................................................51 
9.1.8 Contraception and Pregnancy Avoidance Procedure .............................................51 
9.1.9 Pregnancy ...............................................................................................................52 
9.1.10 Documentation of Subjects who are not Randomized ...........................................52 
9.2 
Monitoring Subject Trial Vaccine Compliance ...........................................................52 
9.3 
Schedule of Observations and Procedures ...................................................................52 
9.3.1 Pre-Vaccination Procedures (Day 1 [M0], Day 90 [M3], and Day 180 [M6]) ......52 
9.3.2 Vaccination Procedures (Day 1 [M0], Day 90 [M3], and Day 180 [M6]) ............53 
9.3.3 Post-Vaccination Procedures (Day 1 [M0], Day 90 [M3], and Day 180 [M6]) ....53 
9.3.4 Clinic Visits after Vaccination (Day 30 [M1], Day 120 [M4], Day 210 [M7], 
and Day 360 [M12])...............................................................................................55 
9.3.5 Final (End of Trial) Visit .......................................................................................56 
9.3.6 Post-Trial Care .......................................................................................................56 
9.4 
Biological Sample Retention and Destruction .............................................................56 
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10.0 
ADVERSE EVENTS .........................................................................................................57 
10.1 
Definitions....................................................................................................................57 
10.1.1 Adverse Events ......................................................................................................57 
10.1.2 Solicited Adverse Events .......................................................................................57 
10.1.3 Medically Attended Adverse Events .....................................................................58 
10.1.4 Serious Adverse Events .........................................................................................59 
10.2 
Causality of Adverse Events ........................................................................................59 
10.2.1 Relationship to Trial Procedures ............................................................................59 
10.2.2 Outcome of Adverse Events ..................................................................................60 
10.3 
Additional Points to Consider for Adverse Events ......................................................60 
10.4 
Procedures ....................................................................................................................61 
10.4.1 Collection and Reporting of Adverse Events.........................................................61 
10.4.2 Collection and Reporting of Solicited Adverse Events .........................................62 
10.4.3 Collection and Reporting of Medically Attended Adverse Events ........................62 
10.4.4 Collection and Reporting of Serious Adverse Events............................................62 
10.5 
Follow-up Procedures ..................................................................................................63 
10.5.1 Adverse Events ......................................................................................................63 
10.5.2 Serious Adverse Events .........................................................................................63 
10.5.3 Safety Reporting to Investigators, Institutional Review Boards or Independent 
Ethics Committees, and Regulatory Authorities ....................................................63 
10.5.4 Post-Trial Events ....................................................................................................64 
11.0 
TRIAL-SPECIFIC REQUIREMENT(S) ...........................................................................65 
11.1 
Data Monitoring Committee ........................................................................................65 
12.0 
DATA HANDLING AND RECORD KEEPING .............................................................66 
12.1 
Electronic Case Report Forms .....................................................................................66 
12.2 
Record Retention .........................................................................................................66 
13.0 
STATISTICAL METHODS ..............................................................................................68 
13.1 
Statistical and Analytical Plans ....................................................................................68 
13.1.1 Analysis Sets ..........................................................................................................68 
13.1.2 Analysis of Demographics and Other Baseline Characteristics ............................68 
13.1.3 Immunogenicity Analysis ......................................................................................68 
13.1.4 Safety Analysis ......................................................................................................69 
13.1.5 Interim Analysis and Criteria for Early Termination .............................................70 
13.2 
Determination of Sample Size .....................................................................................70 
14.0 
QUALITY CONTROL AND QUALITY ASSURANCE .................................................72 
14.1 
Trial-Site Monitoring Visits .........................................................................................72 
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14.2 
Protocol Deviations ......................................................................................................72 
14.3 
Quality Assurance Audits and Regulatory Agency Inspections ..................................72 
15.0 
ETHICAL ASPECTS OF THE TRIAL .............................................................................73 
15.1 
Institutional Review Board and/or Independent Ethics Committee Approval ............73 
15.2 
Subject Information, Informed Consent, and Subject Authorization ..........................74 
15.3 
Subject Confidentiality ................................................................................................75 
15.4 
Publication, Disclosure, and Clinical Trial Registration Policy ..................................75 
15.4.1 Publication and Disclosure ....................................................................................75 
15.4.2 Clinical Trial Registration......................................................................................75 
15.4.3 Clinical Trial Results Disclosure ...........................................................................75 
15.5 
Insurance and Compensation for Injury .......................................................................76 
16.0 
REFERENCES ..................................................................................................................77 
 
LIST OF IN-TEXT TABLES 
Table 1.a 
Contact Information .................................................................................................2 
Table 8.a 
Sponsor-Supplied Vaccines and Placebo ...............................................................45 
Table 10.a 
Solicited Local and Systemic AEs .........................................................................57 
Table 10.b 
Severity Scales for Solicited Safety Parameters ....................................................58 
Table 16.a 
Serology Plan .........................................................................................................85 
 
LIST OF IN-TEXT FIGURES 
Figure 2.a        Schematic of Trial Design .....................................................................................15 
Figure 6.a 
Schematic of Trial Design .....................................................................................35 
 
LIST OF APPENDICES 
Appendix A 
Responsibilities of the Investigator ........................................................................79 
Appendix B 
Investigator Consent to Use of Personal Information ............................................81 
Appendix C 
Elements of the Subject Informed Consent Form ..................................................82 
Appendix D 
Serology Plan .........................................................................................................85 
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2.0 
TRIAL SUMMARY 
Name of Sponsor: 
Takeda Vaccines, Inc. 
40 Landsdowne Street, 
Cambridge, MA 02139, USA 
Product Name: 
Tetravalent Dengue Vaccine Candidate (TDV) 
Trial Title: A Randomized, Observer-Blind, Placebo-Controlled Phase 3 Trial to Investigate the Immunogenicity and 
Safety of a Tetravalent Dengue Vaccine Candidate and a Yellow Fever YF-17D Vaccine Administered Concomitantly 
and Sequentially in Healthy Subjects Aged 18 to 60 years in Non-Endemic Country(ies) 
IND No.: 014292 
EudraCT No.: Not Applicable 
Trial Identifier: DEN-305 
Phase: 3 
Trial Blinding Schema: Observer-Blind 
Background and Rationale: 
Dengue fever is caused by infection with the wild type dengue virus (DENV), a ribonucleic acid virus that occurs as 
4 recognized serotypes, DENV-1, DENV-2, DENV-3, or DENV-4. These dengue viruses are transmitted from human 
to human by mosquitoes (primarily Aedes aegypti). The 4 dengue viruses are endemic in Asia, Central and South 
America, the Caribbean, the Pacific Islands, and parts of Africa. There are an estimated 390 million dengue infections 
per year worldwide, which is more than 3 times the previous World Health Organization (WHO) estimate of 50 to 
100 million cases. Every year, around 500,000 cases of dengue hemorrhagic fever (DHF) require hospitalization with 
an estimated death rate of 2.5%, primarily in children. It is estimated that 3.9 billion people are at risk of dengue 
infection. 
Dengue fever is clinically defined as an acute febrile illness with 2 or more of the following manifestations: headache, 
retro-orbital pain, myalgia, arthralgia, rash, hemorrhagic manifestations, or leucopenia, and occurrence at the same 
location and time as other confirmed cases of dengue fever. The most severe forms of dengue infection – DHF and 
dengue shock syndrome (DSS) – are life threatening. Primary infection with any one of the 4 dengue serotypes is 
thought to result in life-long protection from re-infection by the same serotype, but does not protect against a 
secondary infection by one of the other 3 dengue serotypes and may lead to an increased risk of severe disease 
(DHF/DSS). 
Treatment of dengue fever is based solely on signs and symptoms, with fluid replacement required for hemorrhagic or 
shock cases. An antiviral therapy for dengue virus infection is not available. Preventive measures that rely on 
mosquito control and individual protection are of limited efficacy, complex to implement and questionable in terms of 
cost-effectiveness. There is a great unmet global public health need for a safe and effective vaccine to reduce the 
morbidity and mortality associated with dengue disease. Vaccine development has focused on tetravalent vaccines 
that provide protection against all 4 dengue serotypes simultaneously since all 4 dengue serotypes commonly 
co-circulate in endemic areas. A first tetravalent dengue vaccine (chimeric yellow fever virus-dengue virus tetravalent 
dengue vaccine [CYD-TDV]) has been recently approved in some countries in Asia and Latin America. Initial 
findings showed that vaccine efficacy was different between serotypes and depended on dengue pre-exposure status. 
Additionally, recent analyses found that people who had not been infected by dengue virus before vaccination had a 
higher risk of getting severe disease when they were infected with dengue virus after vaccination with CYD-TDV. 
Hence, there is a continued unmet public health need for safer and more efficacious dengue vaccines. 
Takeda’s Tetravalent Dengue Vaccine Candidate (TDV) - Background: 
Takeda’s TDV consists of 1 molecularly characterized, attenuated dengue serotype 2 virus strain and 3 chimeric 
recombinant dengue virus strains expressing surface antigens corresponding to dengue serotypes 1–4. The dengue 
serotype 2 strain (TDV-2) is based upon the attenuated laboratory-derived virus, DENV-2 Primary Dog Kidney 
(PDK)-53, originally isolated at Mahidol University, Bangkok, Thailand. The chimeric, attenuated vaccine strains for 
dengue serotypes 1, 3 and 4 were engineered by substituting the structural genes, pre-membrane (prM) and envelope 
(E), of TDV-2 with the prM and E genes of the wild type (DENV) virus strains, DENV-1 16007, DENV-3 16562 or 
DENV-4 1036 virus, respectively. Thus, TDV is comprised of 4 dengue virus strains: a molecularly characterized, 
attenuated dengue serotype 2 strain (TDV-2), a dengue serotypes 2/1 chimeric strain (TDV-1), a dengue serotypes 2/3 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 14 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
chimeric strain (TDV-3), and a dengue serotypes 2/4 chimeric strain (TDV-4). 
Non-clinical studies carried out in mice and non-human primates have demonstrated an acceptable safety, 
immunogenicity, and efficacy profile of TDV. Additionally, data from completed phase 1 and phase 2 trials have 
shown satisfactory reactogenicity, safety and immunogenicity profiles of TDV in adults in non-endemic areas as well 
as in adults and children in endemic areas in Asia and Latin America. Ongoing and completed phase 2 trials have 
enabled the selection of a final TDV dose (lyophilized formulation) and a 2-dose vaccination regimen 3 months apart 
by subcutaneous (SC) injection for use in the pivotal program. 
The current Investigator’s Brochure contains additional product information and a more detailed review of pre-clinical 
and clinical trials. 
Rationale for the Proposed Trial: 
Dengue and yellow fever (YF) viruses belong to the same family of Flaviviridae and share antigenic determinants, 
which may result in cross-reacting antibodies. They are both transmitted between humans by mosquitoes (primarily 
Aedes aegypti), and are endemic in tropical areas of Africa and Latin America with a high public health impact. The 
YF-17D vaccine which is based on a live, attenuated viral strain, is the only commercially available YF vaccine 
administered as a single SC injection. The YF-17D vaccine is highly effective (approaching 100%) and generally safe 
with the exception of very rare cases of vaccine-associated neurotropic and viscerotropic disease. Vaccination against 
YF is required for travelers to certain countries in accordance with the International Health Regulations, and is also 
recommended by the WHO for all subjects travelling to areas where there is evidence of persistent or periodic YF 
virus transmission. 
The main purpose of this trial is to assess the immunogenicity and safety of the concomitant administration of YF-17D 
vaccine and TDV in healthy subjects aged 18 to 60 years living in country(ies) non-endemic for both dengue and YF. 
Because broader and enhanced immune responses following vaccination with a dengue vaccine have been reported in 
YF-immune subjects compared to YF non-immune subjects, a secondary purpose of this trial is to assess the 
immunogenicity and safety of the sequential administration of YF-17D vaccine and the 2-dose regimen of TDV. To 
maintain the trial blind, a placebo (0.9% sodium chloride [NaCl] solution) will be used. Approximately 900 subjects 
are planned to be enrolled in this trial. 
This trial will be conducted in accordance with the protocol, the International Council for Harmonization of Technical 
Requirements for Pharmaceuticals for Human Use and Good Clinical Practice (ICH-GCP) Guidelines and applicable 
regulatory requirements. 
Trial Design: 
This is a phase 3, observer-blind, randomized, multi-center trial in 900 healthy adults aged 18 to 60 years in 
non-endemic areas for dengue and YF to investigate the immunogenicity and safety of the concomitant and sequential 
administration of TDV and YF-17D vaccine. Subjects will be randomized equally (1:1:1 ratio) to one of the following 
3 trial groups (300 subjects per trial group): 
– 
Group 1: YF-17D vaccine+placebo concomitantly administered on Day 1 (Month 0 [M0]), first dose of TDV 
administered on Day 90 (Month 3 [M3]), and second dose of TDV administered on Day 180 (Month 6 [M6]). 
– 
Group 2: first dose of TDV+placebo concomitantly administered on Day 1 (M0), second dose of TDV 
administered on Day 90 (M3), and YF-17D vaccine administered on Day 180 (M6). 
– 
Group 3: first dose of TDV+YF-17D vaccine concomitantly administered on Day 1 (M0), second dose of 
TDV administered on Day 90 (M3), and placebo administered on Day 180 (M6). 
Concomitantly administered vaccines will be injected into opposite arms. All subjects will be followed-up for 
6 months after the third vaccination (administered approximately 6 months after the first vaccination), so the trial 
duration will be approximately 360 days (12 months) for each subject. 
A trial population of 900 subjects (300 subjects per trial group) is considered sufficient for the non-inferiority (NI) 
assessment of the immune response to the YF-17D vaccine when concomitantly administered with TDV compared to 
placebo, and for the NI assessment of the immune response to TDV when TDV is concomitantly administered with 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 15 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
YF-17D vaccine compared to placebo. 
Safety parameters include solicited local (injection site) and solicited systemic adverse events (AE) for 7 days (day of 
vaccination+6 days) and 14 days (day of vaccination+13 days) after each vaccination, respectively, unsolicited AEs 
for 28 days (day of vaccination+27 days) after each vaccination, medically attended AEs (MAAE) and serious adverse 
events (SAE) throughout the trial. 
The schematic of the trial design is included as Figure 2.a. 
Figure 2.a        Schematic of Trial Design 
 
Immunogenicity evaluation: 
Blood samples for the measurement of dengue neutralizing antibodies (microneutralization test 50% [MNT50]) will be 
collected at pre-first vaccination (Day 1 [M0]), 1 month post first vaccination (Day 30 [Month 1; M1]), pre-second 
vaccination (Day 90 [M3]), 1 month post second vaccination (Day 120 [Month 4; M4]), pre-third vaccination 
(Day 180 [M6]), and 1 month post third vaccination (Day 210 [Month 7; M7]). 
Blood samples for the measurement of YF neutralizing antibodies (plaque reduction neutralization test [PRNT]) will 
be collected at pre-first vaccination (Day 1 [M0]), 1 month post first vaccination (Day 30 [M1]), pre-third vaccination 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 16 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
(Day 180 [M6]), and 1 month post third vaccination (Day 210 [M7]). 
Safety evaluation: 
 Diary cards (paper or electronic) will be distributed to all subjects for the recording of: 
– 
Solicited local AEs for 7 days following each vaccination (day of vaccination+6 days). These will include: 
injection site pain, injection site erythema, and injection site swelling, and will be collected at each injection 
site. 
– 
Solicited systemic AEs for 14 days following each vaccination (day of vaccination+13 days). These will 
include: fever, headache, asthenia, malaise, and myalgia. 
 Unsolicited AEs for 28 days following each vaccination (day of vaccination+27 days). 
 SAEs and MAAEs will be recorded for the trial duration. MAAEs are defined as AEs leading to an unscheduled 
visit to or by a healthcare professional including visits to an emergency department, but not fulfilling seriousness 
criteria. 
Data collection will be by electronic Case Report Form. 
Primary Objective: 
 To demonstrate NI of the YF seroprotection rate response to 1 dose of YF-17D vaccine, 1 month following 
concomitant administration with 1 dose of TDV compared to placebo. 
Secondary Objectives: 
Immunogenicity 
 To demonstrate NI of the geometric mean titer (GMT) response to TDV for all 4 dengue serotypes, 1 month post 
second dose of TDV following concomitant administration of the first dose of TDV with YF-17D vaccine or 
placebo. 
 To demonstrate NI of the GMT response to 1 dose of YF-17D vaccine, 1 month following concomitant 
administration with 1 dose of TDV compared to placebo. 
 To describe the seropositivity rates for all 4 dengue serotypes, 1 month post second dose of TDV following 
concomitant administration of the first dose of TDV with YF-17D vaccine or placebo. 
 To describe the GMTs of neutralizing antibodies and the seropositivity rates for all 4 dengue serotypes, 1 month 
post first dose of TDV administered concomitantly with YF-17D vaccine or placebo. 
 To describe the GMT and seroprotection rate response to YF-17D vaccine, 1 month after sequential administration 
of a 2-dose regimen of TDV at 0 and 3 months, followed by YF-17D vaccine 3 months later. 
 To describe the GMTs of neutralizing antibodies and the seropositivity rates for all 4 dengue serotypes, 1 month 
after sequential administration of YF-17D vaccine at month 0, followed by a 2-dose regimen of TDV 3 months 
later at 3 and 6 months. 
Safety 
 To assess the safety profile after each vaccine injection in Groups 1, 2 and 3. 
Subject Population: 
Healthy Subjects: Yes 
Planned Minimum Age: 18 years 
Planned Maximum Age: 60 years 
Planned Number of Subjects: 900 
Planned Number of Groups: Three groups in a 1:1:1 ratio (300 subjects in each of Groups 1, 2, and 3) 
Criteria for Inclusion: 
1. The subject is aged 18 to 60 years, inclusive. 
2. Individuals who are in good health at the time of entry into the trial as determined by medical history, physical 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 17 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
examination (including vital signs), and the clinical judgment of the Investigator. 
3. The subject signs and dates a written informed consent form and any required privacy authorization prior to the 
initiation of any trial procedures, and after the nature of the trial has been explained according to local regulatory 
requirements. 
4. Individuals who can comply with trial procedures and are available for the duration of follow-up. 
Criteria for Exclusion: 
1. Individuals with an elevated oral temperature ≥ 38°C (100.4°F) within 3 days of the intended date of vaccination 
(consider whether applicable as an exclusion criterion or criterion for delay, see below). 
2. Known hypersensitivity or allergy to any of the trial vaccine components (including excipients of the 
investigational vaccines and placebo). 
3. Individuals with contraindications, warnings and/or precautions to vaccination with the YF-17D vaccine as 
specified within the product information (especially history of thymus dysfunction). 
4. Individuals with behavioral or cognitive impairment or psychiatric disease that, in the opinion of the Investigator, 
may interfere with the subject’s ability to participate in the trial. 
5. Individuals with any history of progressive or severe neurologic disorder, seizure disorder or neuro-inflammatory 
disease (e.g., Guillain-Barré syndrome). 
6. Individuals with a history of or any current illness that, in the opinion of the Investigator, might interfere with the 
results of the trial or pose additional risk to the subjects due to participation in the trial. 
7. Known or suspected impairment/alteration of immune function, including: 
a) Chronic use of oral steroids (equivalent to 20 mg/day prednisone ≥12 weeks/≥2 mg/kg body weight/day 
prednisone ≥2 weeks) within 60 days prior to Day 1 (M0) (use of inhaled, intranasal, or topical 
corticosteroids is allowed). 
b) Receipt of parenteral steroids (equivalent to 20 mg/day prednisone ≥12 weeks/≥ 2 mg/kg body weight/day 
prednisone ≥2 weeks) within 60 days prior to Day 1 (M0). 
c) Administration of immunoglobulins and/or any blood products within the 3 months prior to Day 1 (M0) or 
planned administration during the trial. 
d) Receipt of immunostimulants within 60 days prior to Day 1 (M0). 
e) Immunosuppressive therapy such as anti-cancer chemotherapy or radiation therapy within 6 months prior to 
Day 1 (M0). 
f) 
Human immunodeficiency virus (HIV) infection or HIV-related disease. 
g) Hepatitis C virus infection. 
h) Genetic immunodeficiency. 
8. Abnormalities of splenic or thymic function. 
9. Individuals with a known bleeding diathesis, or any condition that may be associated with a prolonged bleeding 
time. 
10. Individuals with any serious chronic or progressive disease according to the judgment of the Investigator (e.g., 
neoplasm, insulin dependent diabetes, cardiac, renal or hepatic disease). 
11. Individuals with body mass index (BMI) greater than or equal to 35 kg/m2 (=weight in kg/[height in meters2]). 
12. Individuals or their first degree relatives involved in the trial conduct. 
13. Intent to travel to dengue or YF endemic countries during the trial period. 
14. Individuals with history of substance or alcohol abuse within the past 2 years. 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 18 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
15. Female subjects who are pregnant or breastfeeding. 
16. Any positive or indeterminate pregnancy test. 
17. Females of childbearing potential1 who are sexually active and who have not used any of the acceptable 
contraceptive methods2 for at least 2 months prior to Day 1 (M0). 
18. Females of childbearing potential who are sexually active and who refuse to use an acceptable contraceptive 
method up to 6 weeks post third vaccination on Day 180 (M6). In addition, they must be advised not to donate ova 
during this period.  
19. Individuals participating in any clinical trial with another investigational product 30 days prior to Day 1 (M0) or 
intent to participate in another clinical trial at any time during the conduct of this trial. 
20. Individuals who received any other vaccines within 14 days (for inactivated vaccines) or 28 days (for live 
vaccines) prior to enrollment in this trial or who are planning to receive any non-trial vaccine within 28 days of 
trial vaccine administration. 
21. Previous and planned vaccination (during the trial conduct), against any flavivirus including dengue, YF, 
Japanese encephalitis (JE) or tick-borne encephalitis viruses. 
22. Previous participation in any clinical trial of a dengue or other flavivirus (e.g., West Nile [WN] virus) candidate 
vaccine, except for subjects who received placebo in those trials. 
23. Subjects with a current or previous infection with a flavivirus such as dengue, Zika, YF, JE, WN fever, or Saint 
Louis encephalitis viruses and subjects with a history of prolonged (≥1 year) habitation in a dengue endemic area. 
There may be instances when individuals meet all entry criteria except one that relates to transient clinical 
circumstances (e.g., body temperature elevation or recent use of excluded medication or vaccine). Under these 
circumstances, eligibility for trial enrollment may be considered if the appropriate window for delay has passed, 
inclusion/exclusion criteria have been rechecked, and if the subject is confirmed to be eligible. 
Criteria for delay of second or third vaccination at Day 90 (M3) and Day 180 (M6): 
After enrollment, subjects may encounter clinical circumstances that warrant a delay in the administration of the trial 
vaccination. These situations are listed below. In the event that a subject meets a criterion for delay of vaccination, the 
subject may receive trial vaccination once the window for delay has passed as long as the subject is otherwise eligible 
for trial participation. The decision to vaccinate in those situations will be made by the Investigator. 
1. Body temperature ≥38.0°C (≥100.4°F) within 3 days of intended trial vaccination, and/or use of antipyretics 
and/or analgesic medication for either reason within 24 hours prior to vaccine administration. 
2. Receipt of any vaccine other than the trial vaccines within 14 days (for inactivated vaccines) or 28 days (for 
live vaccines) of intended trial vaccination. 
3. Known or suspected altered or impaired immune function as specified under the exclusion criteria. 
 
Contraindications to vaccination at Day 90 (M3) and Day 180 (M6): 
                                                 
1 Defined as status post onset of menarche and not meeting any of the following conditions: bilateral tubal ligation (at 
least 1 year previously), bilateral oophorectomy (at least 1 year previously) or hysterectomy. 
2 One or more of the following: hormonal contraceptive (such as oral, injection, transdermal patch, implant, cervical 
ring), barrier method (condom with spermicide or diaphragm with spermicide) each and every time during intercourse; 
intrauterine device, monogamous relationship with vasectomized partner (partner must have been vasectomized for at 
least 6 months prior to Day 1 [M0]). Other contraceptive methods may be considered in agreement with the Sponsor 
and will be approved by the appropriate ethics committee. 
 
Property of Takeda: For non-commercial use only and subject to the applicable Terms of Use


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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 20 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
vaccination (Day 210 [M7]) as measured by PRNT. 
Safety 
 
Frequency and severity of solicited local (injection site[s]) AEs for 7 days (day of vaccination+6 days) and 
solicited systemic AEs for 14 days (day of vaccination+13 days) after each vaccination. 
 
Percentage of subjects with any unsolicited AEs within 28 days (day of vaccination+27 days) after each 
vaccination. 
 
Percentage of subjects with MAAEs and SAEs throughout the trial. 
Statistical Considerations: 
Analysis sets 
Safety set: The safety set will consist of all subjects who received at least 1 dose of trial vaccine. 
Full analysis set (FAS): The FAS will include all randomized subjects who received at least 1 dose of trial vaccine and 
for whom a valid pre-dosing and at least 1 post-dosing blood sample have been received for immunogenicity 
assessments. 
Per–protocol set (PPS): The PPS will exclude all subjects seropositive for dengue virus and seroprotected for YF virus 
at Baseline (seropositivity is defined as a reciprocal neutralizing antibody titer ≥10 for the 4 dengue serotypes; YF 
seroprotection is defined as a reciprocal anti-YF neutralizing antibody titer ≥10) and will include all subjects in the 
FAS who have no major protocol violations. The major protocol violation criteria will be defined as part of the blinded 
data review prior to the unblinding of subject’s trial vaccine assignment. The categories of major protocol violations 
include: (1) not meeting selected entry criteria, (2) not receiving the planned trial vaccinations or receiving a wrong 
trial vaccine, (3) receiving prohibited therapies, and (4) other major protocol violations that may be identified during 
blinded data reviews. 
All summaries and analyses of safety data will be based on the safety set. The primary immunogenicity analyses will 
be based on the PPS; additional immunogenicity analyses may be provided based on the FAS. 
Analysis of demographics and other Baseline characteristics 
Age, gender, race, and other Baseline characteristics will be summarized descriptively by trial group for all 
randomized subjects. Unless specified otherwise, number of subjects with non-missing observations, mean, SD, 
median, minimum and maximum will be presented for continuous data; and frequency and percent will be presented 
for categorical data. 
Immunogenicity analysis 
Descriptive statistics including 95% CI for the primary and secondary endpoints, including seroprotection rates, 
seropositivity rates and GMTs, will be computed by trial group for all available assays at all relevant time points. 
As a primary analysis, the NI of the immune response to the YF-17D vaccine when concomitantly administered with 
TDV (Group 3) compared to concomitant administration of the YF-17D vaccine with placebo (Group 1) will be 
assessed in terms of YF seroprotection rates on Day 30 (M1). NI will be concluded if the upper bound of the 95% CI 
for the seroprotection rate difference (Group 1 minus Group 3) is less than the NI margin of 5%. The Newcombe score 
method will be used to compute the 95% CI of the rate difference. 
Secondary analyses will be performed as follows: 
The NI of the immune response to TDV when concomitantly administered with YF-17D vaccine (Group 3) compared 
to concomitant administration of TDV with placebo (Group 2) will be assessed in terms of the GMTs of neutralizing 
antibodies for all 4 dengue serotypes on Day 120 (M4). NI will be concluded if the upper bound of the 95% CI for the 
GMT ratio (Group 2/Group 3) is less than the NI margin of 2.0. An analysis of variance (ANOVA) model will be used 
for this assessment. 
The NI of the immune response to YF-17D vaccine when concomitantly administered with TDV (Group 3) compared 
to concomitant administration of YF-17D vaccine with placebo (Group 1) will be assessed in terms of the GMTs of 
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anti-YF neutralizing antibodies on Day 30 (M1). NI will be concluded if the upper bound of the 95% CI for the GMT 
ratio (Group 1/Group 3) is less than the NI margin of 2.0. An ANOVA model will be used for this assessment. 
Handling of missing data and of values below the lower limit of quantification will be described in the statistical 
analysis plan (SAP). 
Safety analysis 
Solicited AEs 
In all subjects the presence and severity (Grade) of solicited local (injection site[s]) AEs (pain, erythema and swelling) 
and solicited systemic AEs (fever, asthenia, malaise, headache and myalgia) will be collected for 7 days and 14 days 
respectively, following each vaccination (including the day of vaccination) via collection of diary cards. 
For each solicited AE, the number and percentage of subjects with local (injection site[s]) and systemic AEs will be 
summarized by trial group and event severity for each day after each vaccination (i.e., Day 1 through Day 7 for local 
[injection site(s)] AEs and Day 1 through Day 14 for systemic AEs), and overall. Summaries of first onset of each 
event and the number of days subjects reported experiencing each event will also be provided. For subjects with more 
than 1 episode of the same event, the maximum severity will be used for tabulations. 
Persistent/prolonged solicited local or systemic AEs continuing on Day 8 and Day 15, respectively, following each 
trial vaccination will be assessed separately. Unless otherwise specified these AEs will not be included in the 
analyses/tabulations of unsolicited AEs and will have separate listings. 
Unsolicited AEs 
In all subjects, unsolicited AEs will be assessed for 28 days following each vaccination (day of vaccination+27 days). 
Unsolicited AEs will be coded using the Medical Dictionary for Regulatory Activities (MedDRA), and summarized 
by Preferred Term (PT) and System Organ Class (SOC) for each trial group. 
Unsolicited AEs will be summarized as follows: by PT including events with frequency greater than a pre-defined 
frequency (the percentage will be specified in the SAP); by SOC and PT; by SOC, PT, and severity; and by SOC, PT, 
and relationship to the trial vaccine(s). Subjects reporting more than 1 occurrence for the term (level) being 
summarized will be counted only once. 
AEs leading to trial or trial vaccine withdrawal will be collected and summarized for the entire study period. 
MAAEs 
In all subjects, MAAEs will be collected throughout the trial. MAAEs will be coded using MedDRA, and summarized 
by PT and SOC for each trial group. 
SAEs 
In all subjects, SAEs will be collected throughout the trial. SAEs will be coded using MedDRA, and summarized by 
PT and SOC for each trial group. 
Sample Size Justification: 
The immunogenicity set sample size calculation assumes a significance level of 0.025 (one-sided). 
For the primary objective of showing NI in YF seroprotection rates, the calculation assumes a NI margin of 5%, and a 
YF seroprotection rate of 98% at 1 month after YF-17D vaccination in the two trial groups (Group 1 and Group 3). 
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For the secondary objective of showing NI in GMTs of neutralizing antibodies for all the 4 dengue serotypes, the 
calculation assumes a NI margin of 2.0, that the true GMT ratio for the two trial groups (Group 2 and Group 3) is 1, 
and that the natural logarithm of titers are distributed as normal distributions with SDs of 1.35, 0.86, 1.21, and 1.27. 
For the secondary objective of showing NI in GMTs of anti-YF neutralizing antibodies, the calculation assumes a NI 
margin of 2.0, that the true GMT ratio for the two trial groups (Group 1 and Group 3) is 1, and that the natural 
logarithm of titers are distributed as normal distributions with a SD of 1.31. 
A sample size of 300 subjects per trial group, with approximately 255 evaluable subjects per trial group (assuming 
approximately 15% dropouts and non-evaluable subjects), is sufficient to achieve approximately 90% overall power 
for showing NI for the above primary and secondary objectives. A total sample size of 900 subjects also ensures that a 
sufficient number of healthy YF/DENV-naive adults will be vaccinated to support the safe use of TDV in travelers. 
The power calculations were based on nQuery Advisor® 6.01. 
Interim Analysis: 
No interim analyses are planned. 
Data Monitoring Committee: 
A data monitoring committee (DMC) will have oversight of this trial. The DMC functions at a program level and 
further information is available in the DMC Charter. 
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2.1 
Schedule of Trial Procedures 
Visit number 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
Day/Month 
Day 1 
M0 
Day 30 
M1 
Day 90 
M3 
Day 120 
(V3+30 days) 
M4 
Day 180 
(V3+90 days) 
M6 
Day 210 
(V5+30 days) 
M7 
Day 360 
(V5+180 days) 
M12 (ET) (a) 
Visit window (days) 
±0 
-1/+7 
-4/+7 
-1/+7 
-4/+7 
-1/+7 
-7/+14 
Signed informed consent (b) 
X 
 
 
 
 
 
 
Assessment of eligibility criteria (c) 
X 
 
 
 
 
 
 
Demographics (b) 
X 
 
 
 
 
 
 
Medical history (b) 
X 
 
 
 
 
 
 
Concomitant medications/ 
vaccinations (d) 
X 
X 
X 
X 
X 
X 
X 
Check contraindications to vaccination 
 
 
X 
 
X 
 
 
Check criteria for delay of vaccination 
 
 
X 
 
X 
 
 
Complete physical examination (e) 
X 
 
X 
 
X 
 
 
Targeted physical examination (f) 
 
X 
 
X 
 
X 
X 
Vital signs (g) 
X 
X 
X 
X 
X 
X 
X 
Pregnancy test (h) 
X 
 
X 
 
X 
 
 
Pregnancy avoidance guidance (i) 
X 
X 
X 
X 
X 
X 
 
Randomization 
X 
 
 
 
 
 
 
Blood Collection 
YF neutralizing antibodies 
(10 mL) (j) 
X 
X 
 
 
X 
X 
 
Dengue neutralizing 
antibodies (5 mL) (j) 
X 
X 
X 
X 
X 
X 
 
Vaccine administration 
X 
 
X 
 
X 
 
 
30 min post-vaccination in-clinic observation 
including injection site evaluation and body 
temperature measurement (k) 
X 
 
X 
 
X 
 
 
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Visit number 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
Day/Month 
Day 1 
M0 
Day 30 
M1 
Day 90 
M3 
Day 120 
(V3+30 days) 
M4 
Day 180 
(V3+90 days) 
M6 
Day 210 
(V5+30 days) 
M7 
Day 360 
(V5+180 days) 
M12 (ET) (a) 
Visit window (days) 
±0 
-1/+7 
-4/+7 
-1/+7 
-4/+7 
-1/+7 
-7/+14 
Diary Card (l) 
Distribution 
X 
 
X 
 
X 
 
 
Review/collection 
 
X 
 
X 
 
X 
 
Unsolicited AEs (m) 
X 
X 
X 
X 
X 
X 
 
SAEs (n) 
X 
X 
X 
X 
X 
X 
X 
MAAEs (n) 
X 
X 
X 
X 
X 
X 
X 
AEs leading to discontinuation or withdrawal (n) 
X 
X 
X 
X 
X 
X 
X 
Note: AE=adverse event, ET=early termination, M=Month, MAAE=medically attended adverse event, SAE=serious adverse event, V=visit, YF=yellow fever 
Footnotes: 
(a) If the subject terminates early, Day 360 (M12) procedures should be performed. 
(b) Prior to the subject entering into the trial, and before any protocol-directed procedures are performed. 
(c) Review of inclusion/exclusion criteria will be performed prior to the first trial vaccination at Day 1 (M0). After written informed consent has been obtained and eligibility is 
assessed, subjects will be randomized to one of the 3 trial groups. 
(d) Any other vaccination against any flavivirus (licensed or investigational, including any other dengue vaccine) during the entire trial period, all concomitant medications and 
vaccine history from 1 month (minimum 28 days) prior to administration of each dose of trial vaccine(s) up to 1 month (minimum 28 days) thereafter, steroids and 
immunostimulants within 60 days prior to Day 1 (M0), immunoglobulins and blood products within 3 months prior to Day 1 (M0), and immunosuppressive therapy within 6 
months prior to Day 1 (M0). 
(e) Physical examination at Day 1 (M0) including measurement of weight and height; BMI will be calculated automatically. Measurement of height is only required at Day 1 (M0). 
(f) Subjects may undergo a brief symptom-directed physical examination. Clinically significant changes from the Baseline examination should be recorded in the subject’s source 
documents and electronic Case Report Form (eCRF). 
(g) Vital signs including (but not limited to) the measurement of systolic blood pressure/diastolic blood pressure, heart rate, and body temperature. 
(h) Pregnancy testing (serum or urine) for females of childbearing potential. Results must be confirmed and documented as negative prior to each vaccination. Additional pregnancy 
tests may be performed during the trial if deemed necessary by the Investigator. 
(i) Females of childbearing potential who are sexually active will be provided with information on acceptable methods of contraception and will be asked prior to vaccination on 
Day 1 (M0) to sign the informed consent form stating that they understand the requirements for avoidance of pregnancy and donation of ova. Subjects will be reminded during 
trial visits to adhere to acceptable contraceptive methods and not donate ova up to 6 weeks post last trial vaccination at Day 180 (M6). 
(j) The blood sample on Day 1 (M0), Day 90 (M3) and Day 180 (M6) should be taken prior to trial vaccination. The blood sample on Day 120 (M4) and Day 210 (M7) should be 
taken at least 29 days after the vaccination on Day 90 (M3) and Day 180 (M6), respectively. 
(k) After each trial vaccination, the subject will be observed for at least 30 minutes including observation for solicited local (injection site) and systemic AEs, unsolicited AEs 
(non-serious and serious), and body temperature measurement. 
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(l) Diary (paper or electronic) cards will be used for the collection of: 
1) Solicited local (injection site) AEs for 7 days after each vaccination (including the day of vaccination). If solicited local AEs continue on Day 8 following each trial 
vaccination, record the extended information on the Adverse Event eCRF. 
      2) Solicited systemic AEs for 14 days after each vaccination (including the day of vaccination). If solicited systemic AEs continue on Day 15 following each trial vaccination, 
record the extended information on the Adverse Event eCRF. 
The Investigator will categorize events by severity (mild, moderate or severe) and will assess causality to vaccine administration for solicited systemic events (related or not 
related). 
(m) Unsolicited AEs for 28 days (including the day of vaccination) after each vaccination will be collected by interview and recorded for all subjects at Day 30 (M1), Day 120 (M4) 
and Day 210 (M7). The Investigator will categorize events by severity (mild, moderate or severe) and will assess causality to vaccine administration (related or not related). 
(n) AEs leading to discontinuation or withdrawal, MAAEs and SAEs will be collected for the trial duration. The Investigator will categorize events by severity (mild, moderate or 
severe) and will assess causality to vaccine administration (related or not related). 
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3.0 
TRIAL REFERENCE INFORMATION 
3.1 
Trial-Related Responsibilities 
The Sponsor will perform all trial-related activities with the exception of those identified in the 
‘Trial-Related Responsibilities’ template. The identified vendors in the template for specific 
trial-related activities will perform these activities in full or in partnership with the Sponsor. 
3.2 
Principal Investigator/Coordinating Investigator 
The Sponsor will select a Signatory Principal Investigator/Coordinating Investigator from the 
Investigators who participate in the trial. Selection criteria for this Investigator will include 
significant knowledge of the trial protocol, the investigational vaccine, expertise in the therapeutic 
area and the conduct of clinical research as well as trial participation. The Signatory Principal 
Investigator/Coordinating Investigator will be required to review and sign the clinical study report 
and by doing so agrees that it accurately describes the results of the trial. 
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3.3 
List of Abbreviations 
AE 
Adverse Event 
ANOVA 
Analysis Of Variance 
BMI 
Body Mass Index 
CFR 
Code of Federal Regulations 
CYD-TDV 
Chimeric Yellow fever virus-Dengue virus Tetravalent Dengue Vaccine 
DENV 
Wild Type Dengue Virus 
DENV-1, -2, -3, -4 
Wild Type Dengue Virus Serotypes 1, 2, 3, and 4 
DHF 
Dengue Hemorrhagic Fever 
DMC 
Data Monitoring Committee 
DSS 
Dengue Shock Syndrome 
eCRF 
electronic Case Report Form 
EudraCT 
European Clinical Trials Database 
FAS 
Full Analysis Set 
FDA 
Food and Drug Administration 
GCP 
Good Clinical Practice 
GMT 
Geometric Mean Titer 
HIV 
Human Immunodeficiency Virus 
ICH 
International Council for Harmonization of Technical Requirements for 
Pharmaceuticals for Human Use 
IEC 
Independent Ethics Committee 
IND 
Investigational New Drug 
IRB 
Institutional Review Board 
IRT 
Interactive Response Technology 
JE 
Japanese Encephalitis 
M0, 1, 3, 4, 6, 7, 12 
Month 0, 1, 3, 4, 6, 7, 12 
MAAE 
Medically Attended Adverse Event 
MedDRA 
Medical Dictionary for Regulatory Activities 
MNT50 
Microneutralization Test 50% 
NaCl 
Sodium Chloride 
NI 
Non-inferiority 
PPS 
Per–Protocol Set 
PRNT 
Plaque Reduction Neutralization Test 
PT 
Preferred Term 
SAE 
Serious Adverse Event 
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SAP 
Statistical Analysis Plan 
SC 
Subcutaneous 
SOC 
System Organ Class 
SUSAR 
Suspected Unexpected Serious Adverse Reactions 
TDV 
Tetravalent Dengue Vaccine Candidate 
TDV-1 
Dengue serotypes 2/1 chimeric strain 
TDV-2 
Molecularly characterized, attenuated dengue serotype 2 strain 
TDV-3 
Dengue serotypes 2/3 chimeric strain 
TDV-4 
Dengue serotypes 2/4 chimeric strain 
US 
United States 
WHO 
World Health Organization 
WN 
West Nile 
YF 
Yellow Fever 
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3.4 
Corporate Identification 
TV 
Takeda Vaccines, Inc. 
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4.0 
INTRODUCTION 
4.1 
Background 
Dengue fever is caused by infection with the wild type dengue virus (DENV), a ribonucleic acid 
virus that occurs as 4 recognized serotypes, DENV-1, DENV-2, DENV-3, or DENV-4. These 
dengue viruses are transmitted from human to human by mosquitoes (primarily Aedes aegypti). 
The 4 dengue viruses are endemic in Asia, Central and South America, the Caribbean, the Pacific 
Islands, and parts of Africa. There are an estimated 390 million dengue infections per year 
worldwide, which is more than 3 times the previous World Health Organization (WHO) estimate 
of 50 to 100 million cases. Every year, around 500,000 cases of dengue hemorrhagic fever (DHF) 
require hospitalization with an estimated death rate of 2.5%, primarily in children. It is estimated 
that 3.9 billion people are at risk of dengue infection [1-4]. 
Dengue fever is clinically defined as an acute febrile illness with 2 or more of the following 
manifestations: headache, retro-orbital pain, myalgia, arthralgia, rash, hemorrhagic 
manifestations, or leucopenia, and occurrence at the same location and time as other confirmed 
cases of dengue fever. The most severe forms of dengue infection – DHF and dengue shock 
syndrome (DSS) – are life threatening. Primary infection with any one of the 4 dengue serotypes is 
thought to result in life-long protection from re-infection by the same serotype, but does not 
protect against a secondary infection by one of the other 3 dengue serotypes and may lead to an 
increased risk of severe disease (DHF/DSS) [3-6]. 
Treatment of dengue fever is based solely on signs and symptoms, with fluid replacement required 
for hemorrhagic or shock cases. An antiviral therapy for dengue virus infection is not available. 
Preventive measures that rely on mosquito control and individual protection are of limited 
efficacy, complex to implement and questionable in terms of cost-effectiveness. There is a great 
unmet global public health need for a safe and effective vaccine to reduce the morbidity and 
mortality associated with dengue disease [1-7]. Vaccine development has focused on tetravalent 
vaccines that provide protection against all 4 dengue serotypes simultaneously since all 4 dengue 
serotypes commonly co-circulate in endemic areas. A first tetravalent dengue vaccine (chimeric 
yellow fever virus-dengue virus tetravalent dengue vaccine [CYD-TDV]) has been recently 
approved in some countries in Asia and Latin America. [8]. Initial findings showed that vaccine 
efficacy was different between serotypes and depended on dengue pre-exposure status [9]. 
Additionally, recent analyses found that people who had not been infected by dengue virus before 
vaccination had a higher risk of getting severe disease when they were infected with dengue virus 
after vaccination with CYD-TDV [10]. Hence, there is a continued unmet public health need for 
safer and more efficacious dengue vaccines. 
Takeda’s Tetravalent Dengue Vaccine Candidate (TDV) - Background: 
Takeda’s TDV consists of 1 molecularly characterized, attenuated dengue serotype 2 virus strain 
and 3 chimeric recombinant dengue virus strains expressing surface antigens corresponding to 
dengue serotypes 1-4. The dengue serotype 2 strain (TDV-2) is based upon the attenuated 
laboratory-derived virus, DENV-2 Primary Dog Kidney (PDK)-53, originally isolated at Mahidol 
University, Bangkok, Thailand [11]. The chimeric, attenuated vaccine strains for dengue serotypes 
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1, 3 and 4 were engineered by substituting the structural genes, pre-membrane (prM) and envelope 
(E), of TDV-2 with the prM and E genes of the wild type (DENV) virus strains, DENV-1 16007, 
DENV-3 16562 or DENV-4 1036 virus, respectively [12]. Thus TDV is comprised of 4 dengue 
virus strains: a molecularly characterized, attenuated dengue serotype 2 strain (TDV-2), a dengue 
serotypes 2/1 chimeric strain (TDV-1), a dengue serotypes 2/3 chimeric strain (TDV-3), and a 
dengue serotypes 2/4 chimeric strain (TDV- 4). 
Non-clinical studies carried out in mice and non-human primates have demonstrated an acceptable 
safety, immunogenicity, and efficacy profile of TDV. Additionally, data from completed phase 1 
and phase 2 trials have shown satisfactory reactogenicity, safety and immunogenicity profiles of 
TDV in adults in non-endemic areas as well as in adults and children in endemic areas in Asia and 
Latin America. Ongoing and completed phase 2 trials have enabled the selection of a final TDV 
dose (lyophilized formulation) and a 2-dose vaccination regimen 3 months apart by subcutaneous 
(SC) injection for use in the pivotal program. 
The current Investigator’s Brochure for TDV contains additional product information and a more 
detailed review of pre-clinical and clinical trials [13]. 
4.2 
Rationale for the Proposed Trial 
Dengue and yellow fever (YF) viruses belong to the same family of Flaviviridae and share 
antigenic determinants, which may result in cross-reacting antibodies [14]. They are both 
transmitted between humans by mosquitoes (primarily Aedes aegypti), and are endemic in tropical 
areas of Africa and Latin America with a high public health impact. The YF-17D vaccine which is 
based on a live, attenuated viral strain, is the only commercially available YF vaccine administered 
as a single SC injection. The YF-17D vaccine is highly effective (approaching 100%) and 
generally safe with the exception of very rare cases of vaccine-associated neurotropic and 
viscerotropic disease [15]. Vaccination against YF is required for travelers to certain countries in 
accordance with the International Health Regulations [16], and is also recommended by the WHO 
for all subjects travelling to areas where there is evidence of persistent or periodic YF virus 
transmission [15]. 
The main purpose of this trial is to assess the immunogenicity and safety of the concomitant 
administration of YF-17D vaccine and TDV in healthy subjects aged 18 to 60 years living in 
country(ies) non-endemic for both dengue and YF. Because broader and enhanced immune 
responses following vaccination with a dengue vaccine have been reported in YF-immune subjects 
compared to YF non-immune subjects, a secondary purpose of this trial is to assess the 
immunogenicity and safety of the sequential administration of YF-17D vaccine and the 2-dose 
regimen of TDV [17, 18]. To maintain the trial blind, a placebo (0.9% sodium chloride [NaCl] 
solution) will be used. Approximately 900 subjects are planned to be enrolled in the trial. 
This trial will be conducted in accordance with the protocol, the International Council for 
Harmonization of Technical Requirements for Pharmaceuticals for Human Use and Good Clinical 
Practice (ICH-GCP) Guidelines, and applicable regulatory requirements. 
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5.0 
TRIAL OBJECTIVES AND ENDPOINTS 
5.1 
Objectives 
5.1.1 Primary Objective 
 To demonstrate non-inferiority (NI) of the YF seroprotection rate response to 1 dose of 
YF-17D vaccine, 1 month following concomitant administration with 1 dose of TDV 
compared to placebo. 
5.1.2 Secondary Objectives 
Immunogenicity: 
 To demonstrate NI of the geometric mean titer (GMT) response to TDV for all 4 dengue 
serotypes, 1 month post second dose of TDV following concomitant administration of the first 
dose of TDV with YF-17D vaccine or placebo. 
 To demonstrate NI of the GMT response to 1 dose of YF-17D vaccine, 1 month following 
concomitant administration with 1 dose of TDV compared to placebo. 
 To describe the seropositivity rates for all 4 dengue serotypes, 1 month post second dose of 
TDV following concomitant administration of the first dose of TDV with YF-17D vaccine or 
placebo. 
 To describe the GMTs of neutralizing antibodies and the seropositivity rates for all 4 dengue 
serotypes, 1 month post first dose of TDV administered concomitantly with YF-17D vaccine 
or placebo. 
 To describe the GMT and seroprotection rate response to YF-17D vaccine, 1 month after 
sequential administration of a 2-dose regimen of TDV at 0 and 3 months, followed by YF-17D 
vaccine 3 months later. 
 To describe the GMTs of neutralizing antibodies and the seropositivity rates for all 4 dengue 
serotypes, 1 month after sequential administration of YF-17D vaccine at month 0, followed by 
a 2-dose regimen of TDV 3 months later at 3 and 6 months. 
Safety: 
 To assess the safety profile after each vaccine injection in Groups 1, 2 and 3. 
5.2 
Endpoints 
5.2.1 Primary Endpoint 
 Proportion of subjects YF and DENV-naive at Baseline who are seroprotected against YF on 
Day 30 (Month 1 [M1]) as measured by plaque reduction neutralization test (PRNT) (YF 
seroprotection rate). YF seroprotection is defined as reciprocal anti-YF neutralizing antibody 
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titer ≥10. Immunological naivety to YF and DENV is defined as Baseline reciprocal 
neutralizing antibody titers <10 for YF and for the 4 dengue serotypes. 
5.2.2 Secondary Endpoints 
Immunogenicity: 
 GMT of neutralizing antibodies (microneutralization test 50% [MNT50]) for each of the 
4 dengue serotypes at pre-second and -third vaccinations (Day 90 [Month 3; M3] and Day 180 
[Month 6; M6], respectively); and 1 month post first, second, and third vaccinations (Day 30 
[M1], Day 120 [Month 4; M4], and Day 210 [Month 7; M7], respectively) in subjects YF and 
DENV-naive at Baseline. 
 GMTs of anti-YF neutralizing antibodies at 1 month post first and third vaccinations 
(Day 30 [M1] and Day 210 [M7], respectively) in subjects YF and DENV-naive at Baseline. 
 Seropositivity rates (% of subjects seropositive) for each of the 4 dengue serotypes at 
pre-second and -third vaccinations (Day 90 [M3] and Day 180 [M6], respectively); and 
1 month post first, second, and third vaccinations (Day 30 [M1], Day 120 [M4], and Day 210 
[M7], respectively) in subjects YF and DENV-naive at Baseline, where seropositivity is 
defined as a reciprocal neutralizing antibody titer ≥10. 
 Seropositivity rates (% of subjects seropositive) for multiple (2, 3 or 4) dengue serotypes at 
pre-second and -third vaccinations (Day 90 [M3] and Day 180 [M6], respectively); and 
1 month post first, second, and third vaccinations (Day 30 [M1], Day 120 [M4], and Day 210 
[M7], respectively) in subjects YF and DENV-naive at Baseline, where seropositivity is 
defined as a reciprocal neutralizing antibody titer ≥10. 
 Proportion of subjects YF and DENV-naive at Baseline who are seroprotected against YF at 
1 month post third vaccination (Day 210 [M7]) as measured by PRNT. 
Safety: 
 Frequency and severity of solicited local (injection site[s]) Adverse Events (AE) for 7 days 
(day of vaccination+6 days) and solicited systemic AEs for 14 days (day of vaccination+13 
days) after each vaccination. 
 Percentage of subjects with any unsolicited AEs within 28 days (day of vaccination+27 days) 
after each vaccination. 
 Percentage of subjects with Medically Attended AEs (MAAE) and Serious AEs (SAE) 
throughout the trial. 
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6.0 
TRIAL DESIGN AND DESCRIPTION 
6.1 
Trial Design 
This is a phase 3, observer-blind, randomized, multi-center trial in 900 healthy adults aged 18 to 
60 years in non-endemic areas for dengue and YF to investigate the immunogenicity and safety of 
the concomitant and sequential administration of TDV and YF-17D vaccine. Subjects will be 
randomized equally (1:1:1 ratio) to one of the following 3 trial groups (300 subjects per trial 
group): 
 Group 1: YF-17D vaccine+placebo concomitantly administered on Day 1 (Month [M0]), first 
dose of TDV administered on Day 90 (M3) and second dose of TDV administered on 
Day 180 (M6). 
 Group 2: first dose of TDV+placebo concomitantly administered on Day 1 (M0), second dose 
of TDV administered on Day 90 (M3) and YF-17D vaccine administered on Day 180 (M6). 
 Group 3: first dose of TDV+YF-17D vaccine concomitantly administered on Day 1 (M0), 
second dose of TDV administered on Day 90 (M3) and placebo administered on 
Day 180 (M6). 
Concomitantly administered vaccines will be injected into opposite arms. All subjects will be 
followed-up for 6 months after the third vaccination (administered approximately 6 months after 
the first vaccination), so the trial duration will be approximately 360 days (12 months) for each 
subject. 
A trial population of 900 subjects (300 subjects per trial group) is considered sufficient for the NI 
assessment of the immune response to the YF-17D vaccine when concomitantly administered with 
TDV compared to placebo, and for the NI assessment of the immune response to TDV when TDV 
is concomitantly administered with YF-17D vaccine compared to placebo. 
Safety parameters include solicited local (injection site) and solicited systemic AEs for 7 days (day 
of vaccination+6 days) and 14 days (day of vaccination+13 days) after each vaccination, 
respectively, unsolicited AEs for 28 days (day of vaccination+27 days) after each vaccination, 
MAAEs and SAEs throughout the trial. 
A schematic of the trial design is included as Figure 6.a. A schedule of trial procedures is provided 
in Section 2.1. 
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Figure 6.a 
Schematic of Trial Design 
 
Immunogenicity evaluation: 
Blood samples for the measurement of dengue neutralizing antibodies (MNT50) will be collected at 
pre-first vaccination (Day 1 [M0]), 1 month post first vaccination (Day 30 [M1]), pre-second 
vaccination (Day 90 [M3]), 1 month post second vaccination (Day 120 [M4]), pre-third 
vaccination (Day 180 [M6]), and 1 month post third vaccination (Day 210 [M7]). 
Blood samples for the measurement of YF neutralizing antibodies (PRNT) will be collected at 
pre-first vaccination (Day 1 [M0]), 1 month post first vaccination (Day 30 [M1]), pre-third 
vaccination (Day 180 [M6]), and 1 month post third vaccination (Day 210 [M7]). 
Safety evaluation: 
 Diary cards (paper or electronic) will be distributed to all subjects for the recording of: 
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Trial No. DEN-305 
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Protocol Version 4.0 
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 Solicited local AEs for 7 days following each vaccination (day of vaccination+6 days). 
These will include: injection site pain, injection site erythema, and injection site swelling, 
and will be collected at each injection site. 
 Solicited systemic AEs for 14 days following each vaccination (day of 
vaccination+13 days). These will include: fever, headache, asthenia, malaise, and myalgia. 
 Unsolicited AEs for 28 days following each vaccination (day of vaccination+27 days). 
 SAEs and MAAEs will be recorded for the trial duration. MAAEs are defined as AEs leading 
to an unscheduled visit to or by a healthcare professional including visits to an emergency 
department, but not fulfilling seriousness criteria. 
Data collection will be by electronic Case Report Form (eCRF). 
6.2 
Justification for Trial Design, Dose, and Endpoints 
The trial design and the collection of solicited AEs, unsolicited AEs (non-serious AEs and SAEs), 
and MAAEs following vaccination are consistent with vaccine evaluation trials. 
Ongoing and completed phase 2 trials have enabled the selection of a final TDV dose (lyophilized 
formulation) and a 2-dose vaccination regimen 3 months apart by SC injection for use in the 
pivotal program. 
The timing of the primary and secondary endpoints after vaccination is consistent with previous 
trials with TDV. Dengue neutralizing antibodies have been generally accepted as the immune 
response endpoint for dengue vaccine trials. 
The trial is observer-blind. A placebo (0.9% NaCl solution) will be used at the appropriate time 
point to minimize bias. 
As the trial will be conducted in non-endemic areas for dengue, a 6-month follow-up period after 
the third trial vaccination is considered adequate. 
A sample size of 300 subjects per trial group (i.e., 900 subjects in total) is considered sufficient for 
the immunogenicity assessments. The total sample size of 900 subjects is also to ensure that a 
sufficient number of healthy YF/DENV-naive adults will be vaccinated to support the safe use of 
TDV in travelers. Refer to Section 13.2. 
The rationale for the proposed trial is given in Section 4.2. 
6.3 
Duration of Subject’s Expected Participation in the Entire Trial 
The trial duration for each subject will be approximately 360 days (12 months) including 
vaccination (Day 1 [M0], Day 90 [M3] and Day 180 [M6]) and follow-up through 
Day 360 (Month 12 [M12]). 
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Trial No. DEN-305 
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6.4 
Premature Termination or Suspension of Trial or Investigational Site 
6.4.1 Criteria for Premature Termination or Suspension of the Trial 
The trial will be completed as planned unless one or more of the following criteria are satisfied that 
require temporary suspension or early termination of the trial. 
 New information or other evaluation regarding the safety or efficacy of the investigational 
vaccine that indicates a change in the known risk/benefit profile, such that the risk/benefit is no 
longer acceptable for subjects participating in the trial. 
 The data monitoring committee (DMC) recommends that the trial should be suspended or 
terminated. 
 Significant deviation from GCP that compromises the ability to achieve the primary trial 
objectives or compromises subject safety. 
6.4.2 Criteria for Premature Termination or Suspension of Investigational Sites 
A trial site may be terminated prematurely or suspended if the site (including the Investigator) is 
found in significant deviation from GCP, protocol, or contractual agreement, is unable to ensure 
adequate performance of the trial, or as otherwise permitted by the contractual agreement. 
6.4.3 Procedures for Premature Termination or Suspension of the Trial or the 
Participation of Investigational Site(s) 
In the event that the Sponsor, an institutional review board (IRB)/independent ethics committee 
(IEC) or regulatory authority elects to terminate or suspend the trial or the participation of an 
investigational site, a trial-specific procedure for early termination or suspension will be provided 
by the Sponsor; the procedure will be followed by applicable investigational sites during the 
course of termination or trial suspension. 
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Page 38 of 85 
Protocol Version 4.0 
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7.0 
SELECTION AND DISCONTINUATION/WITHDRAWAL OF SUBJECTS 
All entry criteria, including test results, need to be confirmed prior to randomization. 
7.1 
Inclusion Criteria 
Subject eligibility is determined according to the following criteria: 
1. The subject is aged 18 to 60 years, inclusive. 
2. Individuals who are in good health at the time of entry into the trial as determined by medical 
history, physical examination (including vital signs), and the clinical judgment of the 
Investigator. 
3. The subject signs and dates a written informed consent form and any required privacy 
authorization prior to the initiation of any trial procedures, and after the nature of the trial has 
been explained according to local regulatory requirements. 
4. Individuals who can comply with trial procedures and are available for the duration of 
follow-up. 
7.2 
Exclusion Criteria 
Any subject who meets any of the following criteria will not qualify for entry into the trial:  
1. Individuals with an elevated oral temperature ≥ 38°C (100.4°F) within 3 days of the intended 
date of vaccination (consider whether applicable as an exclusion criterion or criterion for 
delay, see below). 
2. Known hypersensitivity or allergy to any of the trial vaccine components (including excipients 
of the investigational vaccines and placebo). 
3. Individuals with contraindications, warnings and/or precautions to vaccination with the 
YF-17D vaccine as specified within the product information (especially history of thymus 
dysfunction). 
4. Individuals with behavioral or cognitive impairment or psychiatric disease that, in the opinion 
of the Investigator, may interfere with the subject’s ability to participate in the trial. 
5. Individuals with any history of progressive or severe neurologic disorder, seizure disorder or 
neuro-inflammatory disease (e.g., Guillain-Barré syndrome). 
6. Individuals with a history of or any current illness that, in the opinion of the Investigator, might 
interfere with the results of the trial or pose additional risk to the subjects due to participation 
in the trial. 
7. Known or suspected impairment/alteration of immune function, including: 
a. Chronic use of oral steroids (equivalent to 20 mg/day prednisone ≥12 weeks/≥2 mg/kg 
body weight/day prednisone ≥2 weeks) within 60 days prior to Day 1 (M0) (use of inhaled, 
intranasal, or topical corticosteroids is allowed). 
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Trial No. DEN-305 
Page 39 of 85 
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b. Receipt of parenteral steroids (equivalent to 20 mg/day prednisone ≥12 weeks/≥2 mg/kg 
body weight/day prednisone ≥2 weeks) within 60 days prior to Day 1 (M0). 
c. Administration of immunoglobulins and/or any blood products within the 3 months prior 
to Day 1 (M0) or planned administration during the trial. 
d. Receipt of immunostimulants within 60 days prior to Day 1 (M0). 
e. Immunosuppressive therapy such as anti-cancer chemotherapy or radiation therapy within 
6 months prior to Day 1 (M0). 
f. Human immunodeficiency virus (HIV) infection or HIV-related disease. 
g. Hepatitis C virus infection. 
h. Genetic immunodeficiency. 
8. Abnormalities of splenic or thymic function. 
9. Individuals with a known bleeding diathesis, or any condition that may be associated with a 
prolonged bleeding time. 
10. Individuals with any serious chronic or progressive disease according to the judgment of the 
Investigator (e.g., neoplasm, insulin dependent diabetes, cardiac, renal or hepatic disease). 
11. Individuals with body mass index (BMI) greater than or equal to 35 kg/m2 (=weight in 
kg/[height in meters2]). 
12. Individuals or their first degree relatives involved in the trial conduct. 
13. Intent to travel to dengue or YF endemic countries during the trial period. 
14. Individuals with history of substance or alcohol abuse within the past 2 years. 
15. Female subjects who are pregnant or breastfeeding. 
16. Any positive or indeterminate pregnancy test. 
17. Females of childbearing potential1 who are sexually active and who have not used any of the 
acceptable contraceptive methods2 for at least 2 months prior to Day 1 (M0). 
18. Females of childbearing potential who are sexually active and who refuse to use an acceptable 
contraceptive method up to 6 weeks post third vaccination on Day 180 (M6). In addition, they 
must be advised not to donate ova during this period. 
                                                 
1 Defined as status post onset of menarche and not meeting any of the following conditions: bilateral tubal ligation (at 
least 1 year previously), bilateral oophorectomy (at least 1 year previously) or hysterectomy. 
2 One or more of the following: hormonal contraceptive (such as oral, injection, transdermal patch, implant, cervical 
ring), barrier method (condom with spermicide or diaphragm with spermicide) each and every time during intercourse; 
intrauterine device, monogamous relationship with vasectomized partner (partner must have been vasectomized for at 
least 6 months prior to Day 1 [M0]). Other contraceptive methods may be considered in agreement with the Sponsor 
and will be approved by the appropriate ethics committee. 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
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Page 40 of 85 
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19. Individuals participating in any clinical trial with another investigational product 30 days prior 
to Day 1 (M0) or intent to participate in another clinical trial at any time during the conduct of 
this trial. 
20. Individuals who received any other vaccines within 14 days (for inactivated vaccines) or 
28 days (for live vaccines) prior to enrollment in this trial or who are planning to receive any 
non-trial vaccine within 28 days of trial vaccine administration. 
21. Previous and planned vaccination (during the trial conduct), against any flavivirus including 
dengue, YF, Japanese encephalitis (JE) or tick-borne encephalitis viruses. 
22. Previous participation in any clinical trial of a dengue or other flavivirus (e.g., West Nile [WN] 
virus) candidate vaccine, except for subjects who received placebo in those trials. 
23. Subjects with a current or previous infection with a flavivirus such as dengue, Zika, YF, JE, 
WN fever, or Saint Louis encephalitis viruses and subjects with a history of prolonged 
(≥1 year) habitation in a dengue endemic area. 
There may be instances when individuals meet all entry criteria except one that relates to transient 
clinical circumstances (e.g., body temperature elevation or recent use of excluded medication or 
vaccine). Under these circumstances, eligibility for trial enrollment may be considered if the 
appropriate window for delay has passed, inclusion/exclusion criteria have been rechecked, and if 
the subject is confirmed to be eligible. 
7.3 
Criteria for Delay of Vaccination and/or Blood Sampling 
After enrollment, subjects may encounter clinical circumstances that warrant a delay in the 
administration of trial vaccination. These situations are listed below. In the event that a subject 
meets a criterion for delay of vaccination, the subject may receive trial vaccination once the 
window for delay has passed as long as the subject is otherwise eligible for trial participation. The 
decision to vaccinate in those situations will be made by the Investigator. 
1. Body temperature ≥38.0°C (≥100.4°F) within 3 days of intended trial vaccination, and/or use 
of antipyretics and/or analgesic medication for either reason within 24 hours prior to vaccine 
administration. 
2. Receipt of any vaccine other than the trial vaccines within 14 days (for inactivated vaccines) or 
28 days (for live vaccines) of intended trial vaccination. 
3. Known or suspected altered or impairment of immune function as specified under the 
exclusion criteria. 
Contraindications to vaccination at Day 90 (M3) and Day 180 (M6): 
There are also circumstances under which receipt of the vaccination at Day 90 (M3) and 
Day 180 (M6) is contraindicated in this trial. These circumstances include but are not limited to 
anaphylaxis or severe hypersensitivity reactions following the administration of first or second 
trial vaccination at Day 1 (M0) or Day 90 (M3), respectively. If these reactions occur, the subject 
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Trial No. DEN-305 
Page 41 of 85 
Protocol Version 4.0 
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must not receive the trial vaccination at Day 90 (M3) or Day 180 (M6), respectively, but is 
encouraged to continue trial participation for safety follow up. 
7.4 
Criteria for Early Trial Termination of a Subject 
Under some circumstances, a subject’s trial participation may be terminated early. This means that 
no further trial procedures (including data collection) will be performed on that subject beyond the 
specific date of early termination. The primary reason for early termination of the subject from the 
trial should be recorded in the eCRF “end of trial visit” page using the following categories. For 
screen failure subjects, refer to Section 9.1.10. 
1. Adverse Event: The subject has experienced an AE (irrespective of being related/unrelated to 
the trial vaccine or trial-related procedures) that requires early termination because continued 
participation imposes an unacceptable risk to the subject’s health and/or the subject is 
unwilling to continue participation because of the AE. If the subject is unwilling to continue 
because of the AE the primary reason for early termination in this case will be ‘withdrawal due 
to AE’ and not ‘withdrawal of consent’, see below. Any ongoing AEs leading to early 
termination should be followed by the Investigator until resolution or stabilization. 
2. Lost to follow-up: The subject did not return to the clinic and at least 3 attempts to contact the 
subject were unsuccessful. Attempts to contact the subject must be documented. 
3. Withdrawal of consent: The subject wishes to withdraw from the trial. The primary reason for 
early termination will be ‘withdrawal of consent” if the subject withdraws from participation 
due to a non-medical reason (i.e., reason other than AE). The reason for withdrawal, if 
provided, should be recorded in the eCRF. 
Note: All attempts should be made to determine the underlying reason for the withdrawal and, 
where possible, the primary underlying reason should be recorded. 
4. Premature trial termination by Sponsor, a regulatory agency, the IEC/IRB, or any other 
authority. 
If the clinical trial is prematurely terminated by the Sponsor, the Investigator is to promptly 
inform the trial subjects and local EC/IRB and should ensure appropriate follow up for the 
subjects. The primary reason for early termination in this case will be ‘trial termination”. 
5. Subject’s death during trial participation. 
6. Other. 
Note: The specific reasons should be recorded in the “specify” field of the eCRF. 
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Trial No. DEN-305 
Page 42 of 85 
Protocol Version 4.0 
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7.5 
Criteria for Premature Discontinuation of Trial Vaccine Administration  
Early trial termination of a subject will by default prevent the subject from continued trial vaccine 
administration, as the subject will no longer be participating in the trial. In addition to early 
termination criteria (see Section 7.4), other situations may apply in which subjects may continue 
participating in the trial (e.g., contributing safety data according to protocol) but trial vaccine 
administration is discontinued selectively. Regardless of the reasons for discontinuation of trial 
vaccine administration, this must be documented as protocol deviation. Even if the subject is 
deemed ineligible to receive trial vaccine, all efforts should be made to continue the collection of 
safety data according to protocol. In addition, the primary reason for premature discontinuation of 
trial vaccine administration should be recorded in the eCRF “end of trial vaccine administration” 
page using the following categories: 
1. Adverse Event: The subject has experienced an AE (irrespective of being related/unrelated to 
the trial vaccine or trial-related procedures) for which subsequent trial vaccine administrations 
impose an unacceptable risk to the subject’s health, but the subject may continue trial 
participation for safety, or a subset of other trial procedures. 
2. Lost to follow-up: The subject did not return to the clinic and at least 3 attempts to contact the 
subject were unsuccessful. Attempts to contact the subject must be documented. 
3. Withdrawal of consent: The subject wishes to withdraw from the trial. The primary reason for 
early termination will be “withdrawal of consent” if the subject withdraws from participation 
due to a non-medical reason (i.e., reason other than AE). The reason for withdrawal, if 
provided, should be recorded in the eCRF. 
4. Premature trial termination by Sponsor, a regulatory agency, the IEC/IRB, or any other 
authority. If the clinical trial is prematurely terminated by the Sponsor, the Investigator is to 
promptly inform the trial subjects and local IEC/IRB and should ensure appropriate follow up 
for the subjects. The primary reason for early termination in this case will be “trial 
termination”. 
5. Subject´s death during trial participation. 
6. Protocol deviation: A protocol deviation is any change, divergence, or departure from the trial 
design or procedures of a trial protocol. The subject may remain in the trial unless continuation 
in the trial jeopardizes the subject’s health, safety or rights (see Section 7.4). 
7. Pregnancy: Any subject who, despite the requirement for adequate contraception, becomes 
pregnant during the trial will not receive further trial vaccine administrations. Pregnant 
subjects should, however, be asked to continue participating in the trial contributing data to the 
safety follow-up according to protocol (i.e., the pregnant subject will not participate in any 
further trial interventions [e.g., blood collection], except for safety follow-up if the subject 
agrees). In addition, the site should maintain contact with the pregnant subject and complete a 
“Pregnancy Form” as soon as possible. The subject should be followed-up until the birth of the 
child, or spontaneous or voluntary termination; when pregnancy outcome information 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
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Protocol Version 4.0 
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becomes available, the information should be captured using the same form. Data obtained 
from the “Pregnancy Form” will be captured in the safety database. 
8. Receipt of any other dengue vaccines (investigational or licensed) during the trial. 
9. Other 
Note: The specific reasons should be recorded in the “specify” field of the eCRF. 
Property of Takeda: For non-commercial use only and subject to the applicable Terms of Use


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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 45 of 85 
Protocol Version 4.0 
28 November 2018 
 
 
The diluent (0.5 mL) is provided in a single-use pre-filled syringe. 
Placebo: 
Commercially available normal saline solution for injection (0.9% NaCl) will be used as placebo. 
The placebo is presented in single dose units for 0.5 mL dosing. 
The Sponsor will supply study sites with packaged and labeled TDV, TDV diluent, YF-17D 
vaccine, YF-17D vaccine diluent and placebo. The labels will contain pertinent trial information in 
local languages. Further details can be found in the Pharmacy Manual. 
8.1.2 Storage 
TDV, YF-17D vaccine, TDV diluent, and YF-17D vaccine diluent will be shipped in refrigerated 
containers at 2°C to 8°C. From receipt and prior to use, TDV, YF-17D vaccine,TDV diluent, and 
YF-17D vaccine diluent must be stored at 2°C to 8°C in a refrigerator, do not freeze. TDV and 
YF-17D vaccine must also be protected from light. 
The placebo will be shipped in ambient conditions and should be stored per the manufacturer’s 
label. 
All clinical trial material must be kept in an appropriate, limited-access, secure place until it is used 
or returned to the Sponsor or designee for destruction. All Sponsor-supplied vaccines must be 
stored under the conditions specified on the label, and remain in the original container until 
dispensed. A daily temperature log of the vaccine storage area must be maintained every working 
day. Temperature excursions must be reported to the Sponsor as soon as possible and use of these 
trial vaccines and diluents requires Sponsor approval. 
8.1.3 Dose and Regimen 
Trial vaccine doses that will be provided to each trial group are presented in Table 8.a. 
The 0.5 mL trial vaccine doses will be prepared and administered by an unblinded pharmacist or 
vaccine administrator according to the instructions in the Pharmacy Manual or per Sponsor 
instructions. 
TDV, YF-17D vaccine, and placebo will be administered by the SC route. 
Table 8.a 
Sponsor-Supplied Vaccines and Placebo 
Trial Group 
Number of subjects 
Day 1 (M0) 
Day 90 (M3) 
Day 180 (M6) 
Group 1 
300 
YF-17D vaccine+placebo 
TDV 
TDV 
Group 2 
300 
TDV+placebo 
TDV 
YF-17D vaccine 
Group 3 
300 
TDV+YF-17D vaccine 
TDV 
Placebo 
Note: Concomitantly administered trial vaccines will be injected in opposite arms. 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
 
Trial No. DEN-305 
Page 46 of 85 
Protocol Version 4.0 
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8.2 
Trial Vaccine Assignment and Dispensing Procedures 
The vaccine to be used will be identifiable by a unique identification number and managed by 
Interactive Response Technology (IRT). Refer to Section 8.6 for accountability of 
Sponsor-supplied vaccines. 
The Investigator or designee will use IRT at subject enrollment to obtain the subject number. This 
number will be used throughout the trial. 
The Investigator or designee will use IRT on the day of first dosing (Day 1 [M0]) to provide the 
necessary subject identifying information. 
The Investigator or designee will use IRT at each dispensing visit to obtain the vaccination 
identification number for the vaccine dose. 
The Investigator’s designee(s) will be responsible for overseeing the administration of vaccine to 
subjects enrolled in the trial according to the procedures stipulated in this trial protocol. The 
vaccine will be administered only by an unblinded pharmacist or vaccine administrator who is 
qualified to perform that function under applicable laws and regulations for that specific trial. 
If Sponsor-supplied vaccine is lost or damaged, the site can request a replacement. Expired 
vaccines must not be administered. 
Prior to vaccination, a subject must be determined to be eligible for trial vaccination and it must be 
clinically appropriate in the judgment of the Investigator to vaccinate. Eligibility for vaccination 
prior to first trial vaccine administration is determined by evaluating the entry criteria outlined in 
this protocol (Section 7.1 and Section 7.2). 
Eligibility for subsequent trial vaccination is determined by the criteria outlined in Section 7.3. 
Trial vaccines should not be administered to individuals with known hypersensitivity to any 
component of the vaccines. 
Standard immunization practices are to be observed and care should be taken to administer the 
injection subcutaneously. In addition, the WHO recommendations to reduce anxiety and pain at 
the time of vaccination should be followed [19]. Before administering the vaccine, the vaccination 
site is to be disinfected with a skin disinfectant (e.g., 70% alcohol). Allow the skin to dry. DO NOT 
inject intravascularly. 
As with all injectable vaccines, trained medical personnel and appropriate medical treatment 
should be readily available in case of anaphylactic reactions following vaccination. For example, 
epinephrine 1:1000, diphenhydramine, and/or other medications for treating anaphylaxis should 
be available. These rescue medications will not be supplied by the Sponsor. 
8.3 
Randomization Code Creation and Storage 
The Sponsor or designee’s randomization personnel will generate the randomization schedule. 
Randomization information will be stored in a secured area, accessible only by authorized 
personnel. 
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8.4 
Trial Vaccine Blind Maintenance 
The trial will be conducted in an observer-blind manner; i.e., during the course of the trial, the 
subject and those responsible for the evaluation of any trial endpoint will all be unaware of which 
trial vaccine(s) was administered. 
The Investigator is responsible for the conduct of the trial according to the protocol, however, they 
will be unaware of which trial vaccine(s) was administered. The unblinded Investigator 
designee(s) will be responsible for preparation, administration, and accountability of trial vaccines 
as specified within the Site Responsibility Delegation Log. The trial vaccines will be prepared and 
administered by an unblinded pharmacist or vaccine administrator according to the instructions in 
the Pharmacy Manual or per Sponsor instructions (see also Section 8.2). The pharmacist (or 
designated individual) at each site will be responsible for vaccine accountability (see also 
Section 8.6). These unblinded designees will maintain the trial vaccine blind and will have no role 
in the assessment of subject safety. 
8.5 
Unblinding Procedure 
The trial vaccine blind shall not be broken by the Investigator unless information concerning the 
trial vaccine(s) is necessary for the medical treatment of the subject or in case of pregnancy if 
requested by the subject. In the event of a medical emergency or pregnancy, if possible, the 
medical monitor should be contacted before the trial vaccine blind is broken to discuss the need for 
unblinding. 
For unblinding a subject, the trial vaccine blind can be obtained by the Investigator, by accessing 
IRT. 
The Sponsor’s Pharmacovigilance Department must be notified as soon as possible if the trial 
vaccine blind is broken by the Investigator and the completed SAE or pregnancy form must be sent 
within 24 hours (Section 10.4.4 and Section 9.1.9, respectively). The date, time, and reason the 
blind is broken must be recorded in the source document and the same information (except the 
time) must be recorded on the eCRF. 
If any subject is unblinded, no further dose(s) of trial vaccine are to be administered and the subject 
must be withdrawn from the trial. 
8.6 
Accountability and Destruction of Sponsor-Supplied Vaccine(s) 
Vaccine supplies will be counted and reconciled at the site before being locally destroyed or 
returned to the Sponsor or designee as noted below. Sites will maintain source documents in 
addition to entering data in IRT. 
The Investigator or designee must ensure that the Sponsor-supplied vaccines (TDV, YF-17D 
vaccine, and placebo) are used in accordance with the approved protocol and are administered only 
to subjects enrolled in the trial. To document appropriate use of Sponsor-supplied vaccines (TDV, 
YF-17D vaccine, and placebo), the Investigator or designee must maintain records of all 
Sponsor-supplied vaccine delivery to the site, site inventory, administration and use by each 
subject, and return to the Sponsor or designee. 
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Upon receipt of Sponsor-supplied vaccines the Investigator or designee must verify the contents of 
the shipments against the packing list. The verifier should ensure that the quantity is correct, the 
vaccines are received within the labeled storage conditions (i.e., no cold chain break has occurred 
during transit), and are in good condition. If quantity and conditions are acceptable, the 
Investigator or designees will record receipt of the shipment in the IRT. 
If there are any discrepancies between the packing list and the actual product received, the Sponsor 
or designee must be contacted to resolve the issue. The packing list should be filed in the 
Investigator’s essential document file by a qualified Investigator designee. 
The pharmacist (or designated individual) at each site must maintain 100% accountability for all 
Sponsor-supplied vaccines received and administered during their entire participation in the trial. 
Proper vaccine accountability includes, but is not limited to: 
 Verifying that the actual inventory matches the documented inventory. 
 Verifying that the log is completed for the vaccine lot number used to prepare each dose. 
 Verifying that all containers used are documented accurately on the log. 
 Verifying that required fields are completed accurately and legibly. 
If any dispensing errors or discrepancies are discovered, the Sponsor must be notified 
immediately. 
The pharmacist (or designated individual) at each site must record the current inventory of all 
Sponsor-supplied vaccines (TDV, YF-17D vaccine, and placebo) including TDV diluent and 
YF-17D vaccine diluent on a Sponsor-approved vaccine accountability log. The following 
information will be recorded at a minimum: protocol number and title, name of Investigator, site 
identifier and number, description of Sponsor-supplied vaccines, expiry date and/or retest date, 
and amount. The log (IRT) should include all required information as a separate entry for each 
subject to whom Sponsor-supplied vaccine is administered. 
Prior to site closure or at appropriate intervals throughout the trial, before any clinical trial 
materials are destroyed locally or returned to the Sponsor or its designee for destruction, a 
representative from the Sponsor or its designee will perform clinical trial material accountability 
and reconciliation. The Investigator will retain a copy of the documentation regarding clinical trial 
material accountability, return, and/or destruction, and originals will be sent to the Sponsor or 
designee. 
The pharmacist (or designated individual) at each site will be responsible for vaccine 
accountability and will document receipt, use, return, or destruction of trial vaccines (TDV, 
YF-17D vaccine, and placebo) including TDV diluent and YF-17D vaccine diluent. 
Vaccine accountability documentation will be reviewed by the unblinded monitor during clinical 
monitoring visits. 
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9.0 
TRIAL PLAN 
9.1 
Trial Procedures 
The following sections describe the trial procedures and data to be collected. For each procedure, 
subjects are to be assessed by the same Investigator or site personnel whenever possible. The 
Schedule of Trial Procedures is presented in Section 2.1. 
9.1.1 Informed Consent Form 
The requirements of the informed consent form are described in Section 15.2. 
Informed consent must be obtained prior to the subject entering into the trial, and before any 
protocol-directed procedures are performed. 
A unique subject number from IRT will be assigned to each subject after informed consent is 
obtained. If all eligibility criteria are fulfilled, this subject number will be used throughout the trial. 
Subject numbers assigned to subjects who fail screening should not be reused (see Section 9.1.10). 
9.1.2 Demographics, Medical History and Prior Medications 
Demographic information to be obtained will include date of birth, sex, race and ethnicity as 
described by the subject. 
Medical history will also be collected, including but not limited to any medical history that may be 
relevant to subject eligibility for trial participation, such as prior medications/vaccinations, 
concomitant medications/vaccinations, and previous and ongoing illnesses or injuries. Relevant 
medical history can also include any medical history that contributes to the understanding of an AE 
that occurs during trial participation, if it represents an exacerbation of an underlying 
disease/pre-existing problem. 
Any other vaccination against any flavivirus (licensed or investigational, including any other 
dengue vaccine) during the entire trial period will be recorded in the eCRF irrespective of time of 
administration, and including the vaccine type. 
All concomitant medications and vaccine history from 1 month (minimum 28 days) prior to 
administration of each dose of trial vaccine(s) up to 1 month (minimum 28 days) thereafter, 
steroids and immunostimulants within 60 days prior to Day 1 (M0), immunoglobulins and blood 
products within 3 months prior to Day 1 (M0), and immunosuppressive therapy within 6 months 
prior to Day 1 (M0) are to be recorded on the “Prior and Concomitant Medications” eCRF page 
and in the subject’s source document. The use of antipyretics and/or analgesic medications within 
24 hours prior to vaccination must be identified and the reason for their use (prophylaxis versus 
treatment) must be described in the source documents or the eCRF. Trial vaccination should be 
delayed if subjects have used antipyretics and/or analgesic medication for either reason within 
24 hours prior to vaccine administration. 
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Medications taken for prophylaxis are those intended to prevent the onset of AEs following 
vaccination. Medications taken for treatment are intended to reduce or eliminate the presence of 
symptoms that are present. 
Prohibited Therapies (see also Section 7.2): 
 Parenteral immunoglobulin preparation, blood products, and/or blood-derived products within 
the 3 months prior to Day 1 (M0). 
 Immunosuppressive therapy within 6 months or systemic (e.g., oral or parenteral) 
corticosteroid treatment within 60 days prior to Day 1 (M0) or immunostimulants within 
60 days prior to Day 1 (M0). 
 Any other vaccines within 14 days (for inactivated vaccines) or 28 days (for live vaccines) 
prior to Day 1 (M0), Day 90 (M3) and Day 180 (M6), and 28 days after each trial vaccination. 
 Any other dengue vaccines (investigational or licensed) for the entire trial period. 
 Receipt of any other clinical trial product for the entire trial period. 
These data must be written in the source documents. 
Medical history (including corresponding medication) to be obtained will include any significant 
conditions or diseases that have disappeared or resolved at or prior to signing of the informed 
consent form. 
9.1.3 Documentation of Trial Entrance/Randomization 
Only subjects who have a signed informed consent form, meet all of the inclusion criteria and none 
of the exclusion criteria are eligible for randomization into the vaccination phase. The 
randomization schedule will be created and controlled by the IRT provider. The randomization 
specification will be approved by the Sponsor’s trial statistician or designee. 
If the subject is found to be ineligible for the randomization/vaccination phase, the Investigator 
should record the primary reason for failure on the subject enrollment log. 
9.1.4 Physical Examination 
Physical examinations must be performed by a qualified health professional in accordance with 
local regulations and licensing requirements designated within the Site Responsibility Delegation 
Log. Complete physical examination will be performed prior to vaccination on Day 1 (M0), 
Day 90 (M3), and Day 180 (M6). A complete physical examination includes but is not limited to: 
auscultation of heart and lungs, palpation of the abdomen, inspection of extremities (including skin 
over intended vaccination site[s]), review of systems, a check of general appearance and the 
measurement of weight and height (measurement of height is only required at Day 1 [M0]); BMI 
will be calculated automatically. Additional physical examinations may be performed if indicated 
by review of the subject’s medical history. The findings should be documented in the subject’s 
source document. 
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Targeted physical examination including but not limited to measurement of vital signs (see 
Section 9.1.5) will be performed at Day 30 (M1), Day 120 (M4), Day 210 (M7) and 
Day 360 (M12). Clinical significant changes from the Baseline assessment must be recorded in the 
subject’s source documents and the eCRF. 
9.1.5 Vital Signs 
These will include (but are not limited to) the measurement of systolic blood pressure, diastolic 
blood pressure, heart rate, and body temperature at all scheduled visits (Day 1 [M0], Day 30[M1], 
Day 90 [M3], Day 120 [M4], Day180 [M6], Day 210 [M7] and Day 360 [M12]). 
9.1.6 Immunogenicity Assessments 
Blood samples for the measurement of dengue neutralizing antibodies (MNT50) will be collected at 
pre-first vaccination (Day 1 [M0]), 1 month post first vaccination (Day 30 [M1]), pre-second 
vaccination (Day 90 [M3]), 1 month post second vaccination (Day 120 [M4]), pre-third 
vaccination (Day 180 [M6]), and 1 month post third vaccination (Day 210 [M7]). Blood samples 
for the measurement of YF neutralizing antibodies (PRNT) will be collected at pre-first 
vaccination (Day 1 [M0]), 1 month post first vaccination (Day 30 [M1]), pre-third vaccination 
(Day 180 [M6]), and 1 month post third vaccination (Day 210 [M7]). The maximum volume of 
blood taken at any single visit is approximately 15 mL, and the approximate total volume of blood 
for the trial is maximum 70 ml. Refer also to Appendix D. 
All samples will be collected in accordance with acceptable laboratory procedures. Blood samples 
will be processed and stored at the trial site according to the Laboratory Guidelines as provided in 
the Laboratory Manual. 
9.1.7 Safety Assessments 
Safety assessments will include collection and recording of solicited local (injection site) and 
systemic AEs, unsolicited AEs (non-serious AEs and SAEs), and MAAEs. Refer to Section 10.1 
for safety definitions. Details on collection and reporting of AEs are in Section 10.4. 
9.1.8 Contraception and Pregnancy Avoidance Procedure 
For female subjects of childbearing potential, serum or urine pregnancy testing will be performed 
prior to vaccination at Day 1 (M0), Day 90 (M3), and Day 180 (M6). Additional pregnancy tests 
may be performed during the trial if deemed necessary by the Investigator. Females of 
childbearing potential who are sexually active will be reminded during trial visits to adhere to 
acceptable contraceptive methods up to 6 weeks post third vaccination at Day 180 (M6). Subjects 
will be provided with information on acceptable methods of contraception and will be asked prior 
to vaccination on Day 1 (M0) to sign the informed consent form stating that they understand the 
requirements for avoidance of pregnancy and donation of ova. During the course of the trial, 
subjects of childbearing potential will receive continued guidance with respect to the avoidance of 
pregnancy and donation of ova as part of the trial procedures (Section 2.1); refer also to 
Section 7.2. 
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9.1.9 Pregnancy 
To ensure subject safety and the safety of the unborn child, each pregnancy in a subject having 
received a trial vaccine must be reported to the Sponsor within 24 hours of the site learning of its 
occurrence. If the subject becomes pregnant during the trial, she will not receive any further doses 
of any Sponsor-supplied trial vaccine. The pregnancy must be followed to determine outcome, 
including spontaneous or voluntary termination, details of birth, and the presence or absence of 
any birth defects, congenital abnormalities, or maternal and/or newborn complications. This 
follow-up should occur even if the intended duration of safety follow-up for the trial has ended. If 
the trial is still ongoing at the time of birth or termination, the women should be further followed 
up for safety until the end of the trial. 
Any pregnancy occurring following trial vaccine administration should be reported immediately, 
using the pregnancy form, to the contact listed in the Investigator Site File. 
The Investigator must inform the subject of their right to receive trial vaccine information. If the 
subject chooses to receive unblinded trial vaccine information, the individual blind should be 
broken by the Investigator and procedures must be followed as described in Section 8.5. 
9.1.10 Documentation of Subjects who are not Randomized 
Investigators must account for all subjects who sign an informed consent form. If the subject is 
found to be not eligible at Day 1 (M0), the Investigator should complete the eCRF. The IRT 
supplier should be contacted as a notification of non-randomization. 
The primary reason for non-randomization is to be recorded in the eCRF using the following 
categories: 
 Screen failure (did not meet one or more inclusion criteria or met one or more exclusion 
criteria). 
 Withdrawal by subject. 
 Trial terminated by Sponsor. 
Subject numbers assigned to subjects who fail screening should not be reused. 
9.2 
Monitoring Subject Trial Vaccine Compliance 
The Investigator records all injections of trial vaccine given to the subject in the eCRF. 
9.3 
Schedule of Observations and Procedures 
The schedule for all trial-related procedures for all evaluations is shown in Section 2.1. 
Assessments should be completed at the designated visit/time point(s). 
9.3.1 Pre-Vaccination Procedures (Day 1 [M0], Day 90 [M3], and Day 180 [M6]) 
1. Before performing any other trial procedure, the signed informed consent needs to be obtained 
(Day 1 [M0]). Refer to Section 9.1.1. 
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2. Check inclusion and exclusion criteria (Day 1 [M0]). Refer to Section 7.1 and Section 7.2. 
3. Collect demographic data (Day 1 [M0]). Refer to Section 9.1.2. 
4. Collect medical history (Day 1 [M0]). Refer to Section 9.1.2. 
5. Collect concomitant medications/vaccinations. Refer to Section 9.1.2. 
6. Perform a complete physical examination. Refer to Section 9.1.4. 
7. Check vital signs. Refer to Section 9.1.5. 
8. Review of systems: Review of systems is a structured interview that queries the subject as to 
any complaints the subject has experienced across each organ system. Refer to Section 9.1.4. 
9. Perform pregnancy testing (serum or urine) for female subjects of childbearing potential. Refer 
to Section 9.1.8 and Section 9.1.9. 
10. Randomize subject (Day 1 [M0]). Refer to Section 9.1.3. 
11. Collect pre-vaccination blood sample(s) from all subjects. Refer to Section 9.1.6. 
Blood should be taken from the subject using an aseptic venipuncture technique for serological 
immunogenicity testing. Refer to the detailed collection and handling procedures outlined in 
the Laboratory Manual. 
9.3.2 Vaccination Procedures (Day 1 [M0], Day 90 [M3], and Day 180 [M6]) 
1. Check criteria for delay of trial vaccination (Day 90 [M3] and Day 180 [M6]). Refer to 
Section 7.3. 
2. Check contraindications to trial vaccination (Day 90 [M3] and Day 180 [M6]). Refer to 
Section 7.3. 
3. Vaccinate the subject. Refer to Section 8.1.3. 
9.3.3 Post-Vaccination Procedures (Day 1 [M0], Day 90 [M3], and Day 180 [M6]) 
1. Careful training of the subject on how to measure solicited local (injection site) AEs and body 
temperature, how to complete the diary card and how often to complete the diary card. 
Training should be directed at the individual(s) who will perform the measurements of local 
AEs and those who will enter the information into the diary card. This individual may or may 
not be the subject, but if a person other than the subject enters information into the diary card, 
this person’s identity must be documented in the trial file and this person must receive training 
on the diary card. Training of the subject on how to measure an injection site AE should be 
performed while the subject is under 30-minute observation after vaccination. 
Diary card instructions must include the following: 
 The subject must understand that timely completion of the diary card on a daily basis is a 
critical component of trial participation. In case a paper diary card is used, the subject 
should also be instructed to write clearly and to complete the diary card in pen. Any 
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corrections to the paper diary card that are performed by the person completing the diary 
card should include a single strikethrough line with a brief explanation for any change and 
be initialed and dated. 
Please note: 
Diary cards will be the only source document allowed for remote collection of solicited local 
(injection site) AEs and systemic AEs (including body temperature measurements). The 
following additional rules apply to the documentation of safety information collected by diary 
card: 
 The diary card should be reviewed with the subject. 
 No corrections or additions to the diary card will be allowed after it is reviewed with the 
Investigator/designee. 
 Any data that are identified as implausible or incorrect, and confirmed by the subject to be 
a transcription error should be corrected by the subject on the diary card (for paper diary 
cards, the correction should include a single strikethrough line and should be initialed and 
dated by the subject). 
 Any blank or illegible fields on the diary card not otherwise corrected as above will be 
missing in the eCRF. 
 The site must enter all readable entries on the paper diary card into the eCRF. 
 Any newly described solicited safety information should be added to the diary card by the 
subject (for paper diary, the new information should be initialed and dated). Any new 
unsolicited safety information would be recorded in the subject’s source document as a 
verbally reported event and therefore captured as an AE and recorded in the Adverse Event 
eCRF. 
 Starting on the day of vaccination, the subject will record specific types of events at the 
injection site (solicited local AEs), any specific generalized symptoms (solicited systemic 
AEs), daily body temperature (any method), any other symptoms or change in the subject’s 
health status, and any medications taken (excluding vitamins and minerals). These 
solicited AEs and body temperature will be recorded on the diary card. Assessments should 
preferably take place in the evening at day’s end. 
 Body temperature measurement is to be performed using the thermometer provided by the 
site. If the subject feels unusually hot or cold during the day, the subject should check their 
temperature. The highest body temperature observed that day should be recorded on the 
diary card. 
 The measurement of solicited local (injection site) AEs (pain, erythema, and swelling) is to 
be performed using the ruler provided by the site. 
2. The collection on the diary card of solicited local (injection site) AEs, and solicited systemic 
AEs (including body temperature) will continue for a total of 7 days and 14 days, respectively, 
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following each trial vaccination. Persistent/prolonged solicited local or systemic AEs 
continuing on Day 8 and Day 15 respectively, following each trial vaccination will be recorded 
in the subject source document as a verbally reported event and therefore captured as an AE 
and recorded in the Adverse Event eCRF. Refer to Section 10.4.2. 
3. Provide pregnancy avoidance counseling. Refer to Section 9.1.8. 
4. Collect and record solicited AEs. Refer to Section 10.1.2. 
5. Collect and record unsolicited AEs. Refer to Section 10.4.1. 
6. Collect and record MAAEs. Refer to Section 10.4.3. 
7. Collect and record SAEs. Refer to Section 10.4.4. 
8. Collect and record AEs leading to subject discontinuation or withdrawal. Refer to 
Section 10.4.1. 
After each trial vaccination, the subject will be observed for at least 30 minutes including 
observation for unsolicited AEs (non-serious and serious), solicited local (injection site) and 
systemic AEs, and body temperature measurement. Information should be recorded in the 
electronic data capture system. The Investigator or delegate will take the opportunity to remind the 
subject how to measure solicited AEs and body temperature as part of this observation period. All 
safety data will be collected in the subject’s source documents. 
The site should schedule the next trial activity with the subject. 
The subject will receive a written reminder of the next planned trial activity. The subject will be 
reminded to complete the diary card daily and to contact the site if there are any questions and to 
contact the site immediately (or as soon as the subject is medically stable) if the subject becomes 
pregnant or has a medical condition that leads to a hospitalization or an emergency room visit or is 
otherwise perceived as serious. All contact details will be provided to the subject. 
9.3.4 Clinic Visits after Vaccination (Day 30 [M1], Day 120 [M4], Day 210 [M7], and 
Day 360 [M12]) 
1. Collect and review the diary card (Day 30 [M1], Day 120 [M4], and Day 210 [M7]). 
2. Collect concomitant medications/vaccinations. Refer to Section 9.1.2. 
3. Perform a targeted physical examination. Refer to Section 9.1.4. 
4. Check vital signs. Refer to Section 9.1.5. 
5. Provide pregnancy avoidance counseling (Day 30 [M1], Day 120 [M4], and Day 210 [M7]). 
Refer to Section 9.1.8. 
6. Record persistent/prolonged solicited AEs (Day 30 [M1], Day 120 [M4], and Day 210 [M7]). 
Refer to Sections 9.3.3 and 10.4.2. 
7. Collect and record unsolicited AEs (Day 30 [M1], Day 120 [M4], and Day 210 [M7]). Refer to 
Section 10.4.1. 
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8. Collect and record MAAEs. Refer to Section 10.4.3. 
9. Collect and record SAEs. Refer to Section 10.4.4. 
10. Collect and record AEs leading to subject discontinuation or withdrawal. Refer to 
Section 10.4.1. 
11. Collect blood sample(s) from all subjects (Day 30 [M1], Day 120 [M4], and Day 210 [M7]). 
Refer to Section 9.1.6. 
Blood should be taken from the subject using an aseptic venipuncture technique for serological 
immunogenicity testing. Refer to the detailed collection and handling procedures outlined in the 
Laboratory Manual. 
The site should schedule the next trial activity with the subject (Day 30 [M1], Day 120 [M4], and 
Day 210 [M7]). 
The subject will receive a written reminder of the next planned trial activity, as applicable. The 
subject will be reminded to complete the diary card daily and to contact the site if there are any 
questions and to contact the site immediately (or as soon as the subject is medically stable) if the 
subject becomes pregnant or has a medical condition that leads to a hospitalization or an 
emergency room visit or is otherwise perceived as serious. All contact details will be provided to 
the subject. 
9.3.5 Final (End of Trial) Visit 
The Final Visit will be performed at Day 360 (M12). If a subject terminates earlier, end of trial 
visit procedures should be performed if possible. For all subjects receiving trial vaccine or 
placebo, the Investigator must complete the End of Trial eCRF page. 
9.3.6 Post-Trial Care 
No post-trial care will be provided. 
9.4 
Biological Sample Retention and Destruction 
In this trial, specimens for immune response testing will be collected as described in Section 9.1.6. 
After blood draw and serum processing, the serum samples will be preserved and retained at a 
central laboratory contracted by the Sponsor for this purpose for up to but no longer than 20 years 
or as required by applicable law. The Sponsor has put into place a system to protect the subject’s 
personal information to ensure optimal confidentiality and defined standard processes for sample 
and data collection, storage, analysis, and destruction. 
Serum samples will be used for the analyses defined in this protocol, but could also, with 
permission from the subject, be used to assess, improve or develop tests related to dengue and 
other disease(s) or the vaccine(s) under trial that will allow more reliable measurement of the 
response to the vaccine(s). 
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10.0 
ADVERSE EVENTS 
10.1 
Definitions 
10.1.1 Adverse Events 
An AE is defined as any untoward medical occurrence in a clinical investigation subject 
administered a trial vaccine; it does not necessarily have to have a causal relationship with trial 
vaccine administration. 
An AE can therefore be any unfavorable and unintended sign (e.g., a clinically significant 
abnormal laboratory finding), symptom, or disease temporally associated with the administration 
of a trial vaccine whether or not it is considered related to the trial vaccine. 
AEs will be graded by the Investigator in the following manner: 
Mild 
Grade 1 
Awareness of symptoms that are easily tolerated, causing minimal discomfort and not 
interfering with everyday activities. Relieved with or without symptomatic treatment. 
Moderate 
Grade 2 
Sufficient discomfort is present to cause interference with normal activity. Only partially 
relieved with symptomatic treatment. 
Severe 
Grade 3 
Extreme distress, causing significant impairment of functioning or incapacitation. Prevents 
normal everyday activities. Not relieved with symptomatic treatment. 
10.1.2 Solicited Adverse Events 
The occurrence of selected indicators of safety as listed in Table 10.a will be measured/collected 
for 7 days (solicited local [injection site] AEs) and 14 days (solicited systemic AEs) following 
each trial vaccination (including the day of vaccination) and will be recorded on the eCRF, as 
applicable. These will be summarized in the final report under the category “solicited AEs” to 
differentiate them from other AEs which were not solicited. Any solicited local or systemic AE 
observed as continuing on Day 8 and Day 15, respectively, following each trial vaccination will be 
recorded as an AE on the Adverse Event eCRF for follow-up. For these persistent/prolonged 
solicited AEs the end date will be captured on the Adverse Event eCRF to permit a separate 
analysis from the unsolicited AEs (see Section 10.4.2). 
Table 10.a 
Solicited Local and Systemic AEs 
Solicited local (injection site) AEs: 
Pain 
Erythema 
Swelling 
Solicited systemic AEs: 
Fever (a) 
Headache 
Asthenia 
Malaise 
Myalgia 
(a) Fever is defined as greater than or equal to 38°C (100.4°F) regardless of method taken [20]. 
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The severity of solicited safety parameters will be assessed as described in Table 10.b. 
Table 10.b 
Severity Scales for Solicited Safety Parameters 
Adverse Event 
Severity grade 
Severity 
Pain at injection site 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity with or without treatment 
3 
Severe: Prevents daily activity with or without treatment 
Erythema at injection 
site (a) 
0 
<25 mm 
1 
Mild: >25 – ≤50 mm 
2 
Moderate: >50–≤100 mm 
3 
Severe: >100 mm 
Swelling at injection 
site (a) 
0 
<25 mm 
1 
Mild: >25 – ≤50 mm  
2 
Moderate: >50-≤100 mm 
3 
Severe: >100 mm 
Headache 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity with or without treatment 
3 
Severe: Prevents normal activity with or without treatment 
Asthenia 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Malaise  
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Myalgia 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Fever (b) 
Record body temperature in °C/°F 
(a) Subjects are to record greatest surface diameter in mm on the diary card. 
(b) Fever is defined as greater than or equal to 38°C (100.4°F) regardless of method taken [20]. 
10.1.3 Medically Attended Adverse Events  
MAAEs are defined as AEs leading to an unscheduled visit to or by a healthcare professional 
including visits to an emergency department, but not fulfilling seriousness criteria. 
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10.1.4 Serious Adverse Events  
An SAE is defined as any untoward medical occurrence that: 
1. Results in DEATH. 
2. Is LIFE THREATENING. 
 The term “life threatening” refers to an event in which the subject was at risk of death at the 
time of the event; it does not refer to an event that hypothetically might have caused death 
if it were more severe. 
3. Requires inpatient HOSPITALIZATION or prolongation of existing hospitalization. 
4. Results in persistent or significant DISABILITY/INCAPACITY. 
5. Leads to a CONGENITAL ANOMALY/BIRTH DEFECT in the offspring of a subject. 
6. Is an IMPORTANT MEDICAL EVENT that satisfies any of the following: 
 May require intervention to prevent items 1 through 5 above. 
 May expose the subject to danger, even though the event is not immediately life 
threatening or fatal or does not result in hospitalization. 
10.2 
Causality of Adverse Events  
Relatedness (causality) to vaccine will also be assessed by the Investigator. The relationship of 
each AE, including solicited systemic AEs (all solicited local AEs are considered as related) to 
trial vaccine(s) will be assessed using the following categories: 
Related: 
There is suspicion that there is a relationship between the trial vaccine(s) and the 
AE (without determining the extent of probability); there is a reasonable 
possibility that the trial vaccine(s) contributed to the AE. 
Not Related: 
There is no suspicion that there is a relationship between the trial vaccine(s) and 
the AE; there are other more likely causes and administration of the trial 
vaccine(s) is not suspected to have contributed to the AE. 
10.2.1 Relationship to Trial Procedures 
Relationship (causality) to trial procedures should be determined for all AEs. 
The relationship should be assessed as “Yes” if the Investigator considers that there is reasonable 
possibility that an event is due to a trial procedure. Otherwise, the relationship should be assessed 
as “No”. 
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10.2.2 Outcome of Adverse Events  
Resolved: 
The subject has fully recovered from the event or the condition has 
returned to the level observed at Baseline. 
Resolving: 
The event is improving but the subject is still not fully recovered. 
Not resolved: 
The event is ongoing at the time of reporting and the subject has still not 
recovered. 
Resolved with 
sequelae: 
As a result of the AE, the subject suffered persistent and significant 
disability/incapacity (e.g., became blind, deaf or paralysed). 
Fatal: 
The subject died due to the event. If the subject died due to other 
circumstances than the event, the outcome of the event per se should be 
stated otherwise (e.g., not resolved or resolving). 
Unknown: 
If outcome is not known or not reported. 
10.3 
Additional Points to Consider for Adverse Events 
An untoward occurrence generally may: 
 Indicate a new diagnosis or unexpected worsening of a pre-existing condition. Intermittent 
events for pre-existing conditions or underlying disease should not be considered as AEs. 
 Necessitate therapeutic intervention. 
 Require an invasive diagnostic procedure. 
 Require vaccine discontinuation or a change in concomitant medication. 
 Be considered unfavorable by the Investigator for any reason. 
Diagnoses vs. signs and symptoms: 
 Each event should be recorded to represent a single diagnosis. Accompanying signs (including 
abnormal laboratory values) or symptoms should NOT be recorded as additional AEs. If a 
diagnosis is unknown, signs or symptoms should be recorded appropriately as AEs. 
Worsening of AEs: 
 If the subject experiences a worsening or complication of an AE after starting administration of 
the trial vaccine(s), the worsening or complication should be recorded as a new AE. 
Investigators should ensure that the AE term recorded captures the change in the condition 
(e.g., “worsening of…”). 
 If the subject experiences a worsening or complication of an AE after any change in trial 
vaccine(s), the worsening or complication should be recorded as a new AE. Investigators 
should ensure that the AE term recorded captures the change in the condition (e.g., “worsening 
of…”). 
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Changes in severity of AEs: 
 If the subject experiences changes in severity of an AE, the event should be captured once with 
the maximum severity recorded. 
Preplanned surgeries or procedures: 
 Preplanned procedures (surgeries or therapies) that were scheduled prior to signing of the 
informed consent form are not considered AEs. Complications resulting from any planned 
surgery should be reported as AEs. 
Elective surgeries or procedures: 
 Elective procedures performed where there is no change in the subject’s medical condition 
should not be recorded as AEs, but should be documented in the subject’s source documents. 
Complications resulting from an elective surgery should be reported as AEs. 
10.4 
Procedures 
10.4.1 Collection and Reporting of Adverse Events  
All AEs, whether considered related with the use of the trial vaccine(s) or not, must be monitored 
until symptoms subside and any abnormal laboratory values have returned to Baseline, or until 
there is a satisfactory explanation for the changes observed, or until death, in which case a full 
pathologist’s report should be supplied, if possible. All findings must be reported on an Adverse 
Event eCRF and on the SAE eCRF form, if necessary (see Section 10.4.2). All findings in subjects 
experiencing AEs must also be reported in the subject’s source documents. Any unsolicited AEs 
for 28 days (day of vaccination+27 days) following each trial vaccination will be collected during 
site visits by interview. AEs leading to withdrawal from the study or discontinuation of trial 
vaccination have to be reported throughout the trial period. 
The following information will be documented for each event: 
 Reported term for the AE. 
 Start and end date. 
 Serious (Y/N). 
 Severity. 
 Investigator’s opinion of the causality (relationship) between the event and administration of 
trial vaccine(s) (“related” or “not related”). 
 Investigator’s opinion of the causality (relationship) to trial procedure(s), including the details 
of the suspected procedure. 
 Action taken with the trial vaccine(s). 
 Outcome of event. 
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10.4.2 Collection and Reporting of Solicited Adverse Events 
The occurrence of selected indicators of safety will be collected using diary cards completed by the 
subjects for 7 days (solicited local [injection site] AEs) and 14 days (solicited systemic AEs) 
following each trial vaccination (including the day of vaccination) and will be recorded on the 
“Local and Systemic AE” eCRF, as applicable. These will be summarized in the final report under 
the category “solicited adverse events” to differentiate them from unsolicited AEs. Any solicited 
local (injection site) or systemic AE observed as continuing on Day 8 and Day 15, respectively, 
following each trial vaccination will be additionally recorded as an AE on the Adverse Event 
eCRF for follow-up. For these persistent/prolonged solicited AEs the end date will be captured on 
the Adverse Event eCRF to permit a separate analysis from the unsolicited AEs. Any solicited 
local (injection site) or systemic AE that resolved before 8 days and 15 days, respectively, 
following each trial vaccination, but which recurs at a later time (i.e., if discontinued), will be 
recorded as an unsolicited AE on the Adverse Event eCRF. 
Any solicited AE that meets any of the following criteria must be entered as an AE on the Adverse 
Event eCRF: 
 Solicited local (injection site) or solicited systemic AEs that lead the subject to withdraw from 
the trial. 
 Solicited local (injection site) or solicited systemic AEs that lead to the subject being 
withdrawn from the trial by the Investigator. 
 Solicited local (injection site) or solicited systemic AEs that otherwise meet the definition of 
an SAE (see Section 10.1.2). 
10.4.3 Collection and Reporting of Medically Attended Adverse Events 
MAAEs occurring from first trial vaccination (Day 1 [M0]) until the end of the trial 
(Day 360 [M12]) will be collected during site visits by interview and must be recorded as an AE 
on the Adverse Event eCRF. 
10.4.4 Collection and Reporting of Serious Adverse Events 
Collection of SAEs will commence from the time that the subject is first administered the trial 
vaccines (Day 1 [M0]). Routine collection of SAEs will continue until the end of the trial 
(Day 360 [M12]). 
SAEs should be reported according to the following procedure: 
A Sponsor SAE form must be completed, in English, and signed by the Investigator immediately 
or within 24 hours of first onset or notification of the event. The information should be completed 
as fully as possible but contain, at a minimum: 
 A short description of the event and the reason why the event is categorized as serious. 
 Subject identification number. 
 Investigator’s name. 
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 Name of the trial vaccine(s) – if no unblinding is necessary, in a blinded way. 
 Causality assessment. 
The SAE form should be transmitted within 24 hours to the attention of the contact(s) in the list 
provided to each site. 
Note: For this study, SAE reporting will be performed via eCRF. Only if the electronic data 
capture system is unavailable, a paper Sponsor SAE form should be completed, signed by the 
investigator and transmitted within 24 hours. The SAE must be entered into the eCRF once access 
to the electronic data capture system is restored. 
10.5 
Follow-up Procedures 
10.5.1 Adverse Events 
All AEs will be monitored until resolution or a stable status is reached or until a formal diagnosis 
can be made or until the end of the trial, whichever occurs first. This could potentially be followed 
outside of this trial or in a planned extension trial. 
10.5.2 Serious Adverse Events 
If information not available at the time of the first report becomes available at a later date, the 
Investigator should complete a follow-up SAE form or provide other written documentation and 
fax it immediately within 24 hours of receipt. Copies of any relevant data from the hospital notes 
(e.g., laboratory tests, discharge summary, post-mortem results) should be sent to the Sponsor. 
All SAEs should be followed up until resolution or permanent outcome of the event or is otherwise 
explained. The timelines and procedure for follow-up reports are the same as those for the initial 
report. 
10.5.3 Safety Reporting to Investigators, Institutional Review Boards or Independent 
Ethics Committees, and Regulatory Authorities 
The Sponsor or designee will be responsible for the reporting of all suspected unexpected serious 
adverse reactions (SUSAR) and any other applicable SAEs to regulatory authorities, investigators 
and IRBs or IECs, as applicable, in accordance with national regulations in the countries where the 
trial is conducted. Relative to the first awareness of the event by/or further provision to the 
Sponsor or Sponsor’s designee, SUSARs will be submitted within 7 days for fatal and 
life-threatening events and 15 days for other SUSARs, unless otherwise required by national 
regulations. The Sponsor will also prepare an expedited report for other safety issues where these 
might materially alter the current benefit-risk assessment of an investigational medicinal product 
or that would be sufficient to consider changes in the trial vaccine administration or in the overall 
conduct of the trial. The investigational site also will forward a copy of all expedited reports to 
their IRB or IEC in accordance with national regulations. 
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10.5.4 Post-Trial Events 
Any SAE that occurs outside of the protocol-specified observation period or after the end of the 
trial but considered to be caused by the trial vaccine(s) must be reported to the Sponsor. These 
SAEs will be processed by the Sponsor’s Pharmacovigilance Department. Instructions for how to 
submit these SAEs will be provided in a handout in the Investigator Site File. 
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11.0 
TRIAL-SPECIFIC REQUIREMENT(S) 
11.1 
Data Monitoring Committee 
A DMC will have oversight of this trial. The DMC functions at a program level and further 
information is available in the DMC Charter. 
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12.0 
DATA HANDLING AND RECORD KEEPING 
The full details of procedures for data handling will be documented in the Data Management Plan. 
Adverse events, medical history, and concurrent conditions will be coded using the Medical 
Dictionary for Regulatory Activities (MedDRA, System Organ Class [SOC], High Level Group 
Term, High Level Term, Low Level Term, Preferred Term [PT], and their corresponding 
descriptive terms. Drugs will be coded using the WHO Drug Dictionary. 
12.1 
Electronic Case Report Forms 
Completed eCRFs are required for each subject who provides a signed informed consent form. 
The Sponsor or its designee will supply investigative sites with access to eCRFs. The Sponsor will 
make arrangements to train appropriate site staff in the use of the eCRF. Electronic CRFs must be 
completed in English. 
After completion of the entry process, computer logic checks will be run to identify items, such as 
inconsistent dates, missing data, and questionable values. Queries may be issued by Sponsor 
personnel (or designees) and will be answered by the site. 
Corrections to eCRFs are recorded in an audit trail that captures the old information, the new 
information, identification of the person making the correction, the date the correction was made, 
and the reason for change. Reasons for significant corrections should additionally be included. 
The Principal Investigator or designee must review the eCRFs for completeness and accuracy and 
must sign and date the appropriate eCRFs as indicated. Furthermore, the Investigator must retain 
full responsibility for the accuracy and authenticity of all data entered on the eCRFs. 
Electronic CRFs will be reviewed for completeness and acceptability at the trial site during 
periodic visits by trial monitors. The Sponsor or its designee will be permitted to review the 
subject’s medical and hospital records pertinent to the trial to ensure accuracy of the eCRFs. The 
completed eCRFs are the sole property of the Sponsor and should not be made available in any 
form to third parties, except for authorized representatives of appropriate governmental health or 
regulatory authorities, without written permission of the Sponsor. 
12.2 
Record Retention 
The Investigator agrees to keep the records stipulated in Section 12.0 and those documents that 
include (but are not limited to) the trial-specific documents, the identification log of all 
participating subjects, medical records. Temporary media such as thermal sensitive paper should 
be copied and certified, source worksheets, all original signed and dated informed consent forms, 
subject authorization forms regarding the use of personal health information (if separate from the 
informed consent form), electronic copy of eCRFs, including the audit trail, and detailed records of 
vaccine disposition to enable evaluations or audits from regulatory authorities, the Sponsor or its 
designees. Furthermore, ICH E6 Section 4.9.5 requires the Investigator to retain essential 
documents specified in ICH E6 (Section 8) until at least 2 years after the last approval of a 
marketing application for a specified vaccine indication being investigated or, if an application is 
not approved, until at least 2 years after the investigation is discontinued and regulatory authorities 
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are notified. In addition, ICH E6 Section 4.9.5 states that the trial records should be retained until 
an amount of time specified by applicable regulatory requirements or for a time specified in the 
Clinical Trial Site Agreement between the Investigator and Sponsor. 
Refer to the Clinical Trial Site Agreement for the Sponsor’s requirements on record retention. The 
Investigator should contact and receive written approval from the Sponsor before disposing of any 
such documents. 
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13.0 
STATISTICAL METHODS 
13.1 
Statistical and Analytical Plans 
A statistical analysis plan (SAP) will be prepared and finalized prior to unblinding of subject’s trial 
vaccine assignment. This document will provide further details regarding the definition of analysis 
variables and analysis methodology to address all trial objectives. 
A blinded data review will be conducted prior to unblinding of subject’s trial vaccine assignment. 
This review will assess the accuracy and completeness of the trial database, subject evaluability, 
and appropriateness of the planned statistical methods. 
13.1.1 Analysis Sets 
Safety set: The safety set will consist of all subjects who received at least 1 dose of trial vaccine. 
Full analysis set (FAS): The FAS will include all randomized subjects who received at least 
1 dose of trial vaccine and for whom a valid pre-dosing and at least 1 post-dosing blood sample 
have been received for immunogenicity assessments. 
Per–protocol set (PPS): The PPS will exclude all subjects seropositive for dengue virus and 
seroprotected for YF virus at Baseline (seropositivity is defined as a reciprocal neutralizing 
antibody titer ≥10 for the 4 dengue serotypes; seroprotection is defined as a reciprocal anti-YF 
neutralizing antibody titer ≥10) and will include all subjects in the FAS who have no major 
protocol violations. The major protocol violation criteria will be defined as part of the blinded data 
review prior to the unblinding of subject’s trial vaccine assignment. The categories of major 
protocol violations include: (1) not meeting selected entry criteria, (2) not receiving the planned 
trial vaccinations or receiving a wrong trial vaccine, (3) receiving prohibited therapies, and 
(4) other major protocol violations that may be identified during blinded data reviews. 
All summaries and analyses of safety data will be based on the safety set. The primary 
immunogenicity analyses will be based on the PPS; additional immunogenicity analyses may be 
provided based on the FAS. 
13.1.2 Analysis of Demographics and Other Baseline Characteristics 
Age, gender, race, and other Baseline characteristics will be summarized descriptively by trial 
group for all randomized subjects. Unless specified otherwise, number of subjects with 
non-missing observations, mean, SD, median, minimum and maximum will be presented for 
continuous data; and frequency and percent will be presented for categorical data. 
13.1.3 Immunogenicity Analysis 
Descriptive statistics including 95% CI for the primary and secondary endpoints, including 
seroprotection rates, seropositivity rates and GMTs, will be computed by trial group for all 
available assays at all relevant time points. 
As a primary analysis, the NI of the immune response to the YF-17D vaccine when concomitantly 
administered with TDV (Group 3) compared to concomitant administration of the YF-17D 
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vaccine with placebo (Group 1) will be assessed in terms of YF seroprotection rates on 
Day 30 (M1). NI will be concluded if the upper bound of the 95% CI for the seroprotection rate 
difference (Group 1 minus Group 3) is less than the NI margin of 5%. The Newcombe score 
method [21] will be used to compute the 95% CI of the rate difference. 
Secondary analyses will be performed as follows: 
The NI of the immune response to TDV when concomitantly administered with YF-17D vaccine 
(Group 3) compared to concomitant administration of TDV with placebo (Group 2) will be 
assessed in terms of the GMTs of neutralizing antibodies for all 4 dengue serotypes on 
Day 120 (M4). NI will be concluded if the upper bound of the 95% CI for the GMT ratio 
(Group 2/Group 3) is less than the NI margin of 2.0. An analysis of variance (ANOVA) model will 
be used for this assessment. 
The NI of the immune response to YF-17D vaccine when concomitantly administered with TDV 
(Group 3) compared to concomitant administration of YF-17D vaccine with placebo (Group 1) 
will be assessed in terms of the GMTs of anti-YF neutralizing antibodies on Day 30 (M1). NI will 
be concluded if the upper bound of the 95% CI for the GMT ratio (Group 1/Group 3) is less than 
the NI margin of 2.0. An ANOVA model will be used for this assessment. 
Handling of missing data and of values below the lower limit of quantification will be described in 
the SAP. 
13.1.4 Safety Analysis 
Solicited AEs 
In all subjects, the presence and severity (Grade) of solicited local (injection site[s]) AEs (pain, 
erythema and swelling) and solicited systemic AEs (fever, asthenia, malaise, headache and 
myalgia) will be collected for 7 days and 14 days respectively, following each vaccination 
(including the day of vaccination) via collection of diary cards. 
For each solicited AE, the number and percentage of subjects with local (injection site[s]) and 
systemic AEs will be summarized by trial group and event severity for each day after each 
vaccination (i.e., Day 1 through Day 7 for local [injection site(s)] AEs and Day 1 through Day 14 
for systemic AEs), and overall. Summaries of first onset of each event and the number of days 
subjects reported experiencing each event will also be provided. For subjects with more than 
1 episode of the same event, the maximum severity will be used for tabulations. 
Persistent/prolonged solicited local or systemic AEs continuing on Day 8 and Day 15, 
respectively, following each trial vaccination will be assessed separately. Unless otherwise 
specified these AEs will not be included in the analyses/tabulations of unsolicited AEs and will 
have separate listings.  
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Unsolicited AEs 
In all subjects, unsolicited AEs will be assessed for 28 days following each vaccination (day of 
vaccination+27 days). 
Unsolicited AEs will be coded using the MedDRA, and summarized by PT and SOC for each trial 
group. 
Unsolicited AEs will be summarized as follows: by PT including events with frequency greater 
than a pre-defined frequency (the percentage will be specified in the SAP); by SOC and PT; by 
SOC, PT, and severity; and by SOC, PT, and relationship to the trial vaccine(s). Subjects reporting 
more than 1 occurrence for the term (level) being summarized will be counted only once. 
AEs leading to trial or trial vaccine withdrawal will be collected and summarized for the entire 
study period. 
MAAEs 
In all subjects, MAAEs will be collected throughout the trial. MAAEs will be coded using 
MedDRA, and summarized by PT and SOC for each trial group. 
SAEs 
In all subjects, SAEs will be collected throughout the trial. SAEs will be coded using MedDRA, 
and summarized by PT and SOC for each trial group. 
13.1.5 Interim Analysis and Criteria for Early Termination 
No interim analyses are planned. 
13.2 
Determination of Sample Size 
The immunogenicity set sample size calculation assumes a significance level of 0.025 (one-sided). 
For the primary objective of showing NI in YF seroprotection rates, the calculation assumes a NI 
margin of 5%, and a YF seroprotection rate of 98% at 1 month after YF-17D vaccination in the two 
trial groups (Group 1 and Group 3). 
For the secondary objective of showing NI in GMTs of neutralizing antibodies for all the 4 dengue 
serotypes, the calculation assumes a NI margin of 2.0, that the true GMT ratio for the two trial 
groups (Group 2 and Group 3) is 1, and that the natural logarithm of titers are distributed as normal 
distributions with SDs of 1.35, 0.86, 1.21, and 1.27. 
For the secondary objective of showing NI in GMTs of anti-YF neutralizing antibodies, the 
calculation assumes a NI margin of 2.0, that the true GMT ratio for the two trial groups (Group 1 
and Group 3) is 1, and that the natural logarithm of titers are distributed as normal distributions 
with a SD of 1.31 [22]. 
A sample size of 300 subjects per trial group, with approximately 255 evaluable subjects per trial 
group (assuming approximately 15% dropouts and non-evaluable subjects), is sufficient to achieve 
approximately 90% overall power for showing NI for the above primary and secondary objectives. 
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A total sample size of 900 subjects also ensures that a sufficient number of healthy 
YF/DENV-naive adults will be vaccinated to support the safe use of TDV in travelers. 
The power calculations were based on nQuery Advisor® 6.01. 
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14.0 
QUALITY CONTROL AND QUALITY ASSURANCE 
14.1 
Trial-Site Monitoring Visits 
Monitoring visits to the trial site will be made periodically during the trial to ensure that all aspects 
of the protocol are followed. Source documents will be reviewed for verification of data recorded 
on the eCRFs. Source documents are defined as original documents, data, and records. The 
Investigator and institution guarantee access to source documents by the Sponsor or its designee 
(Contract Research Organization) and by the IRB or IEC. 
All aspects of the trial and its documentation will be subject to review by the Sponsor or designee 
(as long as blinding is not jeopardized), including but not limited to the Investigator’s Binder, trial 
vaccine, subject medical records, informed consent form documentation, documentation of subject 
authorization to use personal health information (if separate from the informed consent forms), 
and review of eCRFs and associated source documents. It is important that the Investigator and 
other trial personnel are available during the monitoring visits and that sufficient time is devoted to 
the process. 
14.2 
Protocol Deviations 
The Investigator should not deviate from the protocol, except where necessary to eliminate an 
immediate hazard to trial subjects. Should other unexpected circumstances arise that will require 
deviation from protocol-specified procedures, the Investigator should consult with the medical 
monitor (and IRB or IEC, as required) to determine the appropriate course of action. There will be 
no exemptions (a prospective approved deviation) from the inclusion or exclusion criteria. 
14.3 
Quality Assurance Audits and Regulatory Agency Inspections 
The trial site also may be subject to quality assurance audits by the Sponsor or designees. In this 
circumstance, the Sponsor-designated auditor will contact the site in advance to arrange an 
auditing visit. The auditor may ask to visit the facilities where laboratory samples are collected, 
where the vaccine is stored and prepared, and any other facility used during the trial. In addition, 
there is the possibility that this trial may be inspected by regulatory agencies, including those of 
foreign governments (e.g., the Food and Drug Administration [FDA], the United Kingdom 
Medicines and Healthcare Products Regulatory Agency, the Pharmaceuticals and Medical Devices 
Agency of Japan). If the trial site is contacted for an inspection by a regulatory body, the Sponsor 
should be notified immediately. The Investigator and institution guarantee access for quality 
assurance auditors to all trial documents as described in Section 14.1. 
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15.0 
ETHICAL ASPECTS OF THE TRIAL 
This trial will be conducted with the highest respect for the individual participants (i.e., subjects) 
according to the protocol, the ethical principles that have their origin in the Declaration of 
Helsinki, and the ICH Harmonized Tripartite Guideline for GCP. Each Investigator will conduct 
the trial according to applicable local or regional regulatory requirements and align his or her 
conduct in accordance with the “Responsibilities of the Investigator” that are listed in Appendix A. 
The principles of Helsinki are addressed through the protocol and through appendices containing 
requirements for informed consent and Investigator responsibilities. 
15.1 
Institutional Review Board and/or Independent Ethics Committee Approval 
IRBs and IECs must be constituted according to the applicable federal/local requirements of each 
participating region. The Sponsor or designee will require documentation noting all names and 
titles of members who make up the respective IRB or IEC. If any member of the IRB or IEC has 
direct participation in this trial, written notification regarding his or her abstinence from voting 
must also be obtained. Those United States (US) sites unwilling to provide names and titles of all 
members due to privacy and conflict of interest concerns should instead provide a Federal Wide 
Assurance Number or comparable number assigned by the Department of Health and Human 
Services. 
The Sponsor or designee will supply relevant documents for submission to the respective IRB or 
IEC for the protocol’s review and approval. This protocol, the Investigator’s Brochure, a copy of 
the informed consent form, and, if applicable, subject recruitment materials and/or advertisements 
and other documents required by all applicable laws and regulations, must be submitted to a 
central or local IRB or IEC for approval. The IRB’s or IEC’s written approval of the protocol and 
subject informed consent form must be obtained and submitted to the Sponsor or designee before 
commencement of the trial (i.e., before shipment of the Sponsor-supplied vaccine or trial-specific 
screening activity). The IRB or IEC approval must refer to the trial by exact protocol title, number, 
and version date; identify versions of other documents (e.g., informed consent form) reviewed; 
and state the approval date. The Sponsor will notify the site once the Sponsor has confirmed the 
adequacy of site regulatory documentation and, when applicable, the Sponsor has received 
permission from the competent authority to begin the trial. Until the site receives notification no 
protocol activities, including screening may occur. 
Sites must adhere to all requirements stipulated by their respective IRB or IEC. This may include 
notification to the IRB or IEC regarding protocol amendments, updates to the informed consent 
form, recruitment materials intended for viewing by subjects, local safety reporting requirements, 
reports and updates regarding the ongoing review of the trial at intervals specified by the 
respective IRB or IEC, and submission of the Investigator’s final status report to IRB or IEC. All 
IRB and IEC approvals and relevant documentation for these items must be provided to the 
Sponsor or its designee. 
Subject incentives should not exert undue influence for participation. Payments to subjects must 
be approved by the IRB or IEC and Sponsor. 
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15.2 
Subject Information, Informed Consent, and Subject Authorization 
Written consent documents will embody the elements of informed consent as described in the 
Declaration of Helsinki and the ICH Guidelines for GCP and will be in accordance with all 
applicable laws and regulations. The informed consent form, subject authorization form (if 
applicable), and subject information sheet (if applicable) describe the planned and permitted uses, 
transfers, and disclosures of the subject’s personal and personal health information for purposes of 
conducting the trial. The informed consent form and the subject information sheet (if applicable) 
further explain the nature of the trial, its objectives, and potential risks and benefits, as well as the 
date informed consent is given. The informed consent form will detail the requirements of the 
participant and the fact that he or she is free to withdraw at any time without giving a reason and 
without prejudice to his or her further medical care. 
The Investigator is responsible for the preparation, content, and IRB or IEC approval of the 
informed consent form and if applicable, the subject authorization form. The informed consent 
form, subject authorization form (if applicable), and subject information sheet (if applicable) must 
be approved by both the IRB or IEC and the Sponsor prior to use. 
The informed consent form, subject authorization form (if applicable), and subject information 
sheet (if applicable) must be written in a language fully comprehensible to the prospective subject. 
It is the responsibility of the Investigator to explain the detailed elements of the informed consent 
form, subject authorization form (if applicable), and subject information sheet (if applicable) to the 
subject. Information should be given in both oral and written form whenever possible and in the 
manner deemed appropriate by the IRB or IEC. 
The subject must be given ample opportunity to: (1) inquire about details of the trial and (2) decide 
whether or not to participate in the trial. If the subject, determines he or she will participate in the 
trial, then the informed consent form and subject authorization form (if applicable) must be signed 
and dated by the subject at the time of consent and prior to the subject entering into the trial. The 
subject should be instructed to sign using their legal names, not nicknames, using blue or black 
ballpoint ink. The Investigator must also sign and date the informed consent form and subject 
authorization (if applicable) at the time of consent and prior to subject entering into the trial; 
however, the Sponsor may allow a designee of the Investigator to sign to the extent permitted by 
applicable law. 
Once signed, the original informed consent form, subject authorization form (if applicable), and 
subject information sheet (if applicable) will be stored in the Investigator’s site file. The 
Investigator must document the date the subject signs the informed consent form in the subject’s 
medical record and eCRF. Copies of the signed informed consent form, the signed subject 
authorization form (if applicable), and subject information sheet (if applicable) shall be given to 
the subject. 
All revised informed consent forms must be reviewed and signed by the relevant subject in the 
same manner as the original informed consent form. The date the revised consent was obtained 
should be recorded in the subject’s medical record and eCRF, and the subject should receive a 
copy of the revised informed consent form. 
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15.3 
Subject Confidentiality 
The Sponsor and designees affirm and uphold the principle of the subject’s right to protection 
against invasion of privacy. Throughout this trial, a subject’s source data will only be linked to the 
Sponsor’s clinical trial database or documentation via a unique identification number. As 
permitted by all applicable laws and regulations, limited subject attributes, such as sex, age, or date 
of birth, and subject initials may be used to verify the subject and accuracy of the subject’s unique 
identification number. 
To comply with ICH Guidelines for GCP and to verify compliance with this protocol, the Sponsor 
requires the Investigator to permit its monitor or designee’s monitor, representatives from any 
regulatory authority (e.g., FDA, Medicines and Healthcare products Regulatory Agency, 
Pharmaceuticals and Medical Devices Agency), the Sponsor’s designated auditors, and the 
appropriate IRBs and IECs to review the subject’s original medical records (source data or 
documents), including, but not limited to, laboratory test result reports, electrocardiogram (ECG) 
reports, admission and discharge summaries for hospital admissions occurring during a subject’s 
trial participation, and autopsy reports. Access to a subject’s original medical records requires the 
specific authorization of the subject as part of the informed consent form process (see 
Section 15.2). 
Copies of any subject source documents that are provided to the Sponsor must have certain 
personally identifiable information removed (i.e., subject name, address, and other identifier fields 
not collected on the subject’s eCRF). 
15.4 
Publication, Disclosure, and Clinical Trial Registration Policy 
15.4.1 Publication and Disclosure 
The results of this trial are expected to be published in a scientific journal. It is anticipated that 
clinical and laboratory co-investigators will participate in authorship. The order of authorship and 
choice of journal will be proposed by the Sponsor to the Principal Investigator(s), to be eventually 
agreed upon by all authors. The data analysis center for this trial will provide the analyses needed 
for publication. Information regarding this trial will be posted on ClinicalTrials.gov. 
15.4.2 Clinical Trial Registration 
In order to ensure that information on clinical trials reaches the public in a timely manner and to 
comply with applicable law, regulation and guidance, the Sponsor will, at a minimum register all 
clinical trials conducted in subjects that it sponsors anywhere in the world on ClinicalTrials.gov or 
other publicly accessible websites before trial initiation. The Sponsor contact information, along 
with Investigator’s city, country, and recruiting status will be registered and available for public 
viewing. 
15.4.3 Clinical Trial Results Disclosure 
The Sponsor will post the results of this clinical trial, regardless of outcome, on ClinicalTrials.gov 
or other publicly accessible websites, as required by applicable laws and/or regulations. 
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Trial completion corresponds to the date on which the final subject was examined or received an 
intervention for the purposes of final collection of data for the primary outcome, whether the 
clinical trial concluded according to the pre-specified protocol or was terminated. 
15.5 
Insurance and Compensation for Injury 
Each subject in the trial must be insured in accordance with the regulations applicable to the site 
where the subject is participating. If a local underwriter is required, then the Sponsor or Sponsor’s 
designee will obtain clinical trial insurance against the risk of injury to clinical trial subjects. Refer 
to the Clinical Trial Site Agreement regarding the Sponsor’s policy on subject compensation and 
treatment for injury. If the Investigator has questions regarding this policy, he or she should 
contact the Sponsor or Sponsor’s designee. 
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16.0 
REFERENCES 
1. Simmons CP, Farrar JJ, Nguyen VVC, Wills B. Dengue. N Engl J Med 2012;366(15):1423-32. 
2. Gubler DJ. Epidemic dengue/dengue hemorrhagic fever as a public health, social and 
economic problem for the 21st century. Trends Microbiol 2002;10(2):100-3. 
3. Guzman GM, Kouri G. Dengue: an update. Lancet Infect Dis 2002;2(1):33-42. 
4. World Health Organization, 2018. Dengue and severe dengue. Fact Sheet. Available at: 
http://www.who.int/mediacentre/factsheets/fs117/en/ [Accessed 28 November 2018]. 
5. World Health Organization, 1997. Dengue hemorrhagic fever: diagnosis, treatment, 
prevention and control, 2nd Edition. Geneva. Available at: 
http://www.who.int/csr/resources/publications/dengue/Denguepublication/en/ [Accessed 
28 November 2018]. 
6. World Health Organization, 2009. Dengue guidelines for diagnosis, treatment, prevention and 
control. Available at: http://www.who.int/tdr/publications/documents/dengue-diagnosis.pdf 
[Accessed 28 November 2018]. 
7. Halstead S, Deen J. The future of dengue vaccines. Lancet 2002;360(9341):1243-5. 
8. Sanofi Pasteur, 2018. An informational website about dengue and dengue vaccination for 
media. Available from http://www.dengue.info [Accessed 28 November 2018].  
9. Hadinegoro SR, Arredondo-García JL, Capeding MR, Deseda C, Chotpitayasunondh T, 
Dietze R, et al. Efficacy and long-term safety of a dengue vaccine in regions of endemic 
disease. N Engl J Med 2015;373(13):1195-206. 
10. Simmons CP. A candidate dengue vaccine walks a tightrope. N Engl J Med. 2015. 
373(13):1263-4. 
11. Yoksan S, Bhamarapravati N, Halstead SB. Dengue virus vaccine development: study on 
biological markers of uncloned dengue 1-4 viruses serially passaged in primary kidney cells. 
In: TD St. George, BH Kay and J Blok (ed.), Arbovirus research in Australia. Proceedings of 
the 4th Symposium. CSIRO/QIMR, Brisbane, Australia, 1986: 35-8. 
12. Huang CY, Butrapet S, Tsuchiya KR, Bhamarapravati N, Gubler DJ, Kinney RM. Dengue 2 
PDK-53 virus as a chimeric carrier for tetravalent dengue vaccine development. J. Virol 
2003;77(21):11436-47. 
13. Tetravalent Dengue Vaccine Candidate (TDV) Investigator’s Brochure (IND 14292), 
Edition 9, 20 July 2018. 
14. Calisher CH, Karabatsos N, Dalrymple JM, Shope RE, Porterfield JS, Westaway EG, et al. 
Antigenic relationships between flaviviruses as determined by cross-neutralization tests with 
polyclonal antisera. J Gen Virol 1989;70 (Pt 1):37-43. 
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15. World Health Organization. Vaccines and vaccination against yellow fever: WHO Position 
Paper, June 2013-recommendations. Vaccine 2015;33(1):76-7. 
16. Wilder-Smith A, Hill DR, Freedman DO. The revised international health regulations (2005): 
impact on yellow fever vaccination in clinical practice. Am J Trop Med Hyg 
2008;78(3):359-60. 
17. Dorrance WR, Frankel JW, Gordon I, Patterson PR, Schlesinger RW, Winter JW. Clinical and 
serologic response of man to immunization with attenuated dengue and yellow fever viruses. J 
Immunol 1956;77(5): 352–64. 
18. Scott RM, Eckels KH, Bancroft WH, Summers PL, McCown JM, Anderson JH, et al. Dengue 
2 vaccine: dose response in volunteers in relation to yellow fever immune status. J Infect Dis 
1983;148 (6):1055–60. 
19. World Health Organization. Reducing pain at the time of vaccination: WHO position paper–
September 2015. Wkly Epidemiol Rec. 2015;90(39):505–10. 
20. Kohl KS, Marcy M, Blum M, Jones MC, Dagan R, Hansen J, et al. Fever after immunization: 
current concepts and improved scientific understanding. Clin Infect Dis 2004;39:389-94. 
21. Newcombe RG. Improved confidence intervals for the difference between binomial 
proportions based on paired data. Stat Med 1998;17(22):2635-50.  
22. Nasveld PE, Marjason J, Bennett S, Aaskov J, Elliott S, McCarthy K, et al. Concomitant or 
sequential administration of live attenuated Japanese encephalitis chimeric virus vaccine and 
yellow fever 17D vaccine: randomized double-blind phase II evaluation of safety and 
immunogenicity. Hum Vaccin 2010;6(11):906-14. 
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APPENDIX A 
RESPONSIBILITIES OF THE INVESTIGATOR 
Clinical research trials sponsored by the Sponsor are subject to ICH GCP and all the applicable 
local laws and regulations. 
The Investigator agrees to assume the following responsibilities: 
1. Conduct the trial in accordance with the protocol. 
2. Personally conduct or supervise the staff who will assist in the protocol. 
3. Ensure that trial-related procedures, including trial-specific (non-routine/non-standard panel) 
screening assessments are NOT performed on potential subjects, prior to the receipt of written 
approval from relevant governing bodies/authorities. 
4. Ensure that all colleagues and employees assisting in the conduct of the trial are informed of 
these obligations. 
5. Secure prior approval of the trial and any changes by an appropriate IRB/IEC that conform to 
21 Code of Federal Regulations (CFR) Part 56, ICH, and local regulatory requirements. 
6. Ensure that the IRB/IEC will be responsible for initial review, continuing review, and approval 
of the protocol. Promptly report to the IRB/IEC all changes in research activity and all 
anticipated risks to subjects. Make at least yearly reports on the progress of the trial to the 
IRB/IEC, and issue a final report within 3 months of trial completion. 
7. Ensure that requirements for informed consent, as outlined in 21 CFR Part 56, ICH and local 
regulations, are met. 
8. Obtain valid informed consent from each subject who participates in the trial, and document 
the date of consent in the subject’s medical chart. Valid informed consent form is the most 
current version approved by the IRB/IEC. Each informed consent form should contain a 
subject authorization Section that describes the uses and disclosures of a subject’s personal 
information (including personal health information) that will take place in connection with the 
trial. If an informed consent form does not include such a subject authorization, then the 
Investigator must obtain a separate subject authorization form from each subject. 
9. Prepare and maintain adequate case histories of all persons entered into the trial, including 
eCRFs, hospital records, laboratory results, etc, and maintain these data for a minimum of 
2 years following notification by the Sponsor that all investigations have been discontinued or 
that the regulatory authority has approved the marketing application. The Investigator should 
contact and receive written approval from the Sponsor before disposing of any such 
documents. 
10. Allow possible inspection and copying by the regulatory authority of GCP-specified essential 
documents. 
11. Maintain current records of the receipt, administration, and disposition of Sponsor-supplied 
vaccines, and return all unused Sponsor-supplied vaccines to the Sponsor. 
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12. Report AEs to the Sponsor promptly. In the event of an SAE, notify the Sponsor within 
24 hours. 
13. Review and provide a signature as approval of the content of the clinical study report. 
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APPENDIX B 
INVESTIGATOR CONSENT TO USE OF PERSONAL 
INFORMATION 
Takeda will collect and retain personal information of Investigator, including his or her name, 
address, and other personally identifiable information. In addition, Investigator’s personal 
information may be transferred to other parties located in countries throughout the world (e.g., the 
United Kingdom, US, and Japan), including the following: 
 Takeda, its affiliates, and licensing partners. 
 Business partners assisting Takeda, its affiliates, and licensing partners. 
 Regulatory agencies and other health authorities. 
 IRBs and IECs. 
Investigator’s personal information may be retained, processed, and transferred by Takeda and 
these other parties for research purposes including the following: 
 Assessment of the suitability of Investigator for the trial and/or other clinical trials. 
 Management, monitoring, inspection, and audit of the trial. 
 Analysis, review, and verification of the trial results. 
 Safety reporting and pharmacovigilance relating to the trial. 
 Preparation and submission of regulatory filings, correspondence, and communications to 
regulatory agencies relating to the trial. 
 Preparation and submission of regulatory filings, correspondence, and communications to 
regulatory agencies relating to other vaccines used in other clinical trials that may contain the 
same chemical compound present in the investigational vaccine. 
 Inspections and investigations by regulatory authorities relating to the trial. 
 Self-inspection and internal audit within Takeda, its affiliates, and licensing partners. 
 Archiving and audit of trial records. 
 Posting Investigator site contact information, trial details and results on publicly accessible 
clinical trial registries, databases, and websites. 
Investigator’s personal information may be transferred to other countries that do not have data 
protection laws that offer the same level of protection as data protection laws in Investigator’s own 
country. Investigator acknowledges and consents to the use of his or her personal information by 
Takeda and other parties for the purposes described above. 
Please note that this consent will not cover Investigator personnel. 
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APPENDIX C 
ELEMENTS OF THE SUBJECT INFORMED CONSENT FORM 
In seeking informed consent, the following information shall be provided to each subject: 
1. A statement that the trial involves research. 
2. An explanation of the purposes of the research. 
3. The expected duration of the subject’s participation. 
4. A description of the procedures to be followed, including invasive procedures. 
5. The identification of any procedures that are experimental. 
6. The estimated number of subjects involved in the trial. 
7. A description of the subject’s responsibilities. 
8. A description of the conduct of the trial. 
9. A statement describing the vaccination(s) and the probability for random assignment to each 
trial group. 
10. A description of the possible side effects following vaccine administration that the subject may 
receive. 
11. A description of any reasonably foreseeable risks or discomforts to the subject and, when 
applicable, to an embryo, fetus, or nursing infant. 
12. A statement that the vaccinations may involve risks to the subject (or to the embryo or fetus, if 
the subject is or may become pregnant) which are currently unforeseeable. 
13. A description of any benefits to the subject or to others that reasonably may be expected from 
the research. When there is no intended clinical benefit to the subject, the subject should be 
made aware of this. 
14. Disclosures of appropriate alternative procedures or courses of treatment, if any, that might be 
advantageous to the subject and their important potential risks and benefits. 
15. A statement describing the extent to which confidentiality of records identifying the subject 
will be maintained, and a note of the possibility that regulatory agencies, auditor(s), IRB/IEC, 
and the monitor may inspect the records. By signing a written informed consent form, the 
subject is authorizing such access. 
16. For research involving more than minimal risk, an explanation as to whether any compensation 
and an explanation as to whether any medical treatments are available if injury occurs and, if 
so, what they consist of or where further information may be obtained. 
17. The anticipated prorated payment(s), if any, to the subject for participating in the trial. 
18. The anticipated expenses, if any, to the subject for participating in the trial. 
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19. An explanation of whom to contact for answers to pertinent questions about the research 
(Investigator), subject’s rights, and IRB/IEC and whom to contact in the event of a 
research-related injury to the subject. 
20. A statement that participation is voluntary, that refusal to participate will involve no penalty or 
loss of benefits to which the subject otherwise is entitled, and that the subject may discontinue 
participation at any time without penalty or loss of benefits to which the subject is otherwise 
entitled. 
21. The consequences of a subject’s decision to withdraw from the research and procedures for 
orderly termination of participation by the subject. 
22. A statement that the subject will be informed in a timely manner if information becomes 
available that may be relevant to the subject’s willingness to continue participation in the trial. 
23. A statement that results of pharmacogenomic analysis will not be disclosed to an individual, 
unless prevailing laws require the Sponsor to do so. 
24. The foreseeable circumstances or reasons under which the subject’s participation in the trial 
may be terminated. 
25. A written subject authorization (either contained within the informed consent form or provided 
as a separate document) describing to the subject the contemplated and permissible uses and 
disclosures of the subject’s personal information (including personal health information) for 
purposes of conducting the trial. The subject authorization must contain the following 
statements regarding the uses and disclosures of the subject’s personal information: 
a) that personal information (including personal health information) may be processed by or 
transferred to other parties in other countries for clinical research and safety reporting 
purposes, including, without limitation, to the following: (1) Takeda, its affiliates, and 
licensing partners; (2) business partners assisting Takeda, its affiliates, and licensing 
partners; (3) regulatory agencies and other health authorities; and (4) IRBs/IECs; 
b) it is possible that personal information (including personal health information) may be 
processed and transferred to countries that do not have data protection laws that offer 
subjects the same level of protection as the data protection laws within this country; 
however, Takeda will make every effort to keep your personal information confidential, 
and your name will not be disclosed outside the clinic unless required by law; 
c) that personal information (including personal health information) may be added to 
Takeda’s research databases for purposes of developing a better understanding of the 
safety and effectiveness of the investigational vaccine(s), studying other therapies for 
patients, developing a better understanding of disease, and improving the efficiency of 
future clinical trials; 
d) that subjects agree not to restrict the use and disclosure of their personal information 
(including personal health information) upon withdrawal from the trial to the extent that 
the restricted use or disclosure of such information may impact the scientific integrity of 
the research; and 
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e) that the subject’s identity will remain confidential in the event that trial results are 
published. 
26. Female subjects of childbearing potential (e.g., non-sterilized, premenopausal female subjects) 
who are sexually active must use adequate contraception (as defined in the informed consent 
form) from at least 2 months prior to Day 1 (M0) up to 6 weeks post third vaccination on 
Day 180 (M6). Pregnancy tests will be performed prior to vaccination at Day 1 (M0), Day 90 
(M3), and Day 180 (M6). Additional pregnancy tests may be performed during the trial if 
deemed necessary by the Investigator. Females of childbearing potential who are sexually 
active will be reminded during trial visits to adhere to acceptable contraceptive methods up to 
6 weeks post third vaccination on Day 180 (M6). In addition, they must be advised not to 
donate ova during this period. If a subject is found to be pregnant during the trial, no further 
vaccine doses will be administered. 
27. A statement that clinical trial information from this trial will be publicly disclosed in a publicly 
accessible website, such as ClinicalTrials.gov. 
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APPENDIX D 
SEROLOGY PLAN 
Table 16.a 
Serology Plan 
Timing 
Blood volume 
Assessments 
Day 1 (M0), Day 30 (M1), Day180 (M6), 
Day 210 days (M7) 
10 mL 
YF neutralizing antibodies (PRNT) 
Day 1 (M0), Day 30 (M1), Day 90 (M3), 
Day 120 (M4), Day180 (M6) and Day 210 (M7) 
5 mL 
Dengue neutralizing antibodies (MNT50) 
M=month, MNT50=microneutralization test 50%, PRNT=plaque reduction neutralization test. 
Note: Blood sampling at Day 1 (M0), Day 90 (M3) and Day 180 (M6) should be performed prior to vaccination. 
Note: The blood sample on Day 120 (M4) and Day 210 (M7) should be taken at least 29 days after the vaccination on 
Day 90 (M3) and Day 180 (M6), respectively. 
Property of Takeda: For non-commercial use only and subject to the applicable Terms of Use


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SAP_001.pdf: 


Other information as needed to protect confidentiality of Takeda or partners, personal
information, or to otherwise protect the integrity of the clinical study.
Title: A Randomized, Observer-Blind, Placebo-Controlled, Phase 3 Trial to Investigate the 
Immunogenicity and Safety of a Tetravalent Dengue Vaccine Candidate and a Yellow Fever 
YF-17D Vaccine Administered Concomitantly and Sequentially in Healthy Subjects 
Aged 18 to 60 years in Non-Endemic Country(ies) 
NCT Number: NCT03342898
SAP Approve Date: 
Certain information within this Statistical Analysis Plan has been redacted (ie, specific content is masked 
irreversibly from view with a black/blue bar) to protect either personally identifiable (PPD) information 
or company confidential information (CCI). 
This may include, but is not limited to, redaction of the following: 
Named persons or organizations associated with the study.
Proprietary information, such as scales or coding systems, which are considered confidential 
information under prior agreements with license holder.


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Takeda’s Tetravalent Dengue Vaccine Candidate 
Trial No. DEN-305 
Page 2 of 34
Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
1.1 
Approval Signatures 
Electronic signatures can be found on the last page of this document. 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
2.0 
TABLE OF CONTENTS 
1.0 
TITLE PAGE .......................................................................................................................1 
1.1 
Approval Signatures.......................................................................................................2 
2.0 
TABLE OF CONTENTS .....................................................................................................3 
3.0 
LIST OF ABBREVIATIONS ..............................................................................................5 
4.0 
OBJECTIVES ......................................................................................................................6 
4.1 
Primary Objective ..........................................................................................................6 
4.2 
Secondary Objectives.....................................................................................................6 
4.3 
Additional Objectives ....................................................................................................6 
4.4 
Study Design ..................................................................................................................6 
5.0 
ANALYSIS ENDPOINTS ................................................................................................10 
5.1 
Primary Endpoints .......................................................................................................10 
5.2 
Secondary Endpoints ...................................................................................................10 
6.0 
DETERMINATION OF SAMPLE SIZE ..........................................................................11 
7.0 
METHODS OF ANALYSIS AND PRESENTATION .....................................................12 
7.1 
General Principles ........................................................................................................12 
7.1.1 Data Presentation ...................................................................................................12 
7.1.2 Study Day, Baseline and Analysis Window Definitions .......................................12 
7.1.3 Handling of Missing Data ......................................................................................14 
7.1.4 Implausible Values.................................................................................................15 
7.2 
Analysis Sets ................................................................................................................15 
7.3 
Disposition of Subjects ................................................................................................17 
7.4 
Demographic and Other Baseline Characteristics .......................................................18 
7.5 
Medical History and Concurrent Medical Conditions .................................................18 
7.6 
Medication History and Concomitant Medications .....................................................19 
7.7 
Investigational Product Exposure and Compliance .....................................................19 
7.8 
Efficacy Analysis .........................................................................................................20 
7.9 
Pharmacokinetic/Pharmacodynamic Analysis .............................................................20 
7.10 
Other Outcomes ...........................................................................................................20 
7.10.1 Primary Immunogenicity Endpoint........................................................................23 
7.10.2 Secondary Immunogenicity Endpoints ..................................................................23 
7.11 
Safety Analysis ............................................................................................................24 
7.11.1 Adverse Events ......................................................................................................24 
7.11.2 Clinical Laboratory Evaluations ............................................................................28 
7.11.3 Vital Signs ..............................................................................................................28 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
7.11.4 12-Lead ECGs ........................................................................................................28 
7.11.5 Other Observations Related to Safety ....................................................................28 
7.12 
Changes in the Statistical Analysis Plan ......................................................................28 
8.0 
REFERENCES ..................................................................................................................29 
 
LIST OF IN-TEXT TABLES 
Table 7.a 
Analysis Visit Windows ........................................................................................13 
Table 7.b 
Plausible Data Ranges............................................................................................15 
Table 7.c 
Criteria for Exclusion from the PPS ......................................................................17 
Table 7.d 
Planned Analyses and Populations for Immunogenicity Endpoints ......................22 
Table 7.e 
Summaries of Solicited Local AEs following Vaccination 1 ................................25 
Table 7.f 
Overview of Unsolicited Adverse Events ..............................................................28 
Table 8.a 
Schedule of Trial Procedures .................................................................................30 
Table 8.b 
Solicited Local (Injection Site) and Systemic AEs ................................................33 
Table 8.c 
Severity of Solicited Safety Parameters .................................................................34 
 
LIST OF IN-TEXT FIGURES 
Figure 4.a 
Schematic of Trial Design .......................................................................................8 
LIST OF APPENDICES 
Appendix A 
Schedule of Trial Procedures .................................................................................30 
Appendix B 
Solicited Local (Injection Site) and Systemic Adverse Events and Severity ........33 
 
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Trial No. DEN-305 
Page 5 of 34
Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
3.0 
LIST OF ABBREVIATIONS 
AE 
Adverse event 
ANOVA 
Analysis of variance 
CRF 
Case Report Form 
FAS 
Full Analysis Set 
GMT 
Geometric mean titer 
ICH 
International Conference on Harmonization 
IP 
Investigational product 
LLOQ 
Lower limit of quantification 
LS means 
Least squares means 
M0, 1, 3, 4, 6, 7, 12 
Month 0, 1, 3, 4, 6, 7, 12 
MAAE 
Medically attended adverse event 
MedDRA 
Medical Dictionary for Regulatory Activities 
MNT50 
Microneutralization test 50% 
NI 
Non-Inferiority 
PPS 
Per-Protocol Set 
PRNT 
Plaque reduction neutralization test 
PT 
Preferred term 
SAE 
Serious adverse event 
SAP 
Statistical analysis plan 
SAS 
Statistical analysis system 
SOC 
System organ class 
TDV 
Tetravalent dengue vaccine candidate 
WHO Drug 
World Health Organization Drug Dictionary 
YF 
Yellow Fever 
 
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Trial No. DEN-305 
Page 6 of 34
Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
4.0 
OBJECTIVES 
4.1 
Primary Objective 
 To demonstrate non-inferiority (NI) of the Yellow Fever (YF) seroprotection rate response to 
1 dose of YF-17D vaccine, 1 month following concomitant administration with 1 dose of a 
tetravalent dengue vaccine candidate (TDV) compared to placebo. 
4.2 
Secondary Objectives 
Immunogenicity: 
 To demonstrate NI of the geometric mean titer (GMT) response to TDV for all 4 dengue 
serotypes, 1-month post second dose of TDV following concomitant administration of the 
first dose of TDV with YF-17D vaccine or placebo. 
 To demonstrate NI of the GMT response to 1-dose of YF-17D vaccine, 1 month following 
concomitant administration with 1 dose of TDV compared to placebo. 
 To describe the seropositivity rates for all 4 dengue serotypes, 1-month post second dose of 
TDV following concomitant administration of the first dose of TDV with YF-17D vaccine or 
placebo. 
 To describe the GMTs of neutralizing antibodies and the seropositivity rates for all 4 dengue 
serotypes, 1-month post first dose of TDV administered concomitantly with YF-17D vaccine 
or placebo. 
 To describe the GMT and seroprotection rate response to YF-17D vaccine, 1 month after 
sequential administration of a 2-dose regimen of TDV at 0 and 3 months, followed by YF-
17D vaccine 3 months later. 
 To describe the GMTs of neutralizing antibodies and the seropositivity rates for all 4 dengue 
serotypes, 1 month after sequential administration of YF-17D vaccine at month 0, followed 
by a 2-dose regimen of TDV 3 months later at 3 and 6 months. 
Safety: 
 To assess the safety profile after each vaccine injection in Groups 1, 2 and 3. 
4.3 
Additional Objectives 
Not applicable. 
4.4 
Study Design 
This is a phase 3, observer-blind, randomized, multi-center trial in 900 healthy adults aged 18 to 
60 years in non-endemic areas for dengue and YF to investigate the immunogenicity and safety 
of the concomitant and sequential administration of TDV and YF-17D vaccine [1]. Subjects will 
be randomized equally (1:1:1 ratio) to one of the following 3 trial groups (300 subjects per trial 
group) using an interactive response technology: 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
 Group 1: YF-17D vaccine + placebo concomitantly administered on Day 1 (Month [M0]), 
first dose of TDV administered on Day 90 (M3) and second dose of TDV administered on 
Day 180 (M6). 
 Group 2: first dose of TDV + placebo concomitantly administered on Day 1 (M0), second 
dose of TDV administered on Day 90 (M3) and YF-17D vaccine administered on 
Day 180 (M6). 
 Group 3: first dose of TDV + YF-17D vaccine concomitantly administered on Day 1 (M0), 
second dose of TDV administered on Day 90 (M3) and placebo administered on 
Day 180 (M6). 
Concomitantly administered vaccines will be injected into opposite arms. All subjects will be 
followed-up for 6 months after the third vaccination (administered approximately 6 months after 
the first vaccination), so the trial duration will be approximately 360 days (12 months) for each 
subject. 
The total trial population of 900 subjects (300 subjects per trial group) is considered sufficient 
for the NI assessment of the immune response to the YF-17D vaccine when concomitantly 
administered with TDV compared to placebo, and for the NI assessment of the immune response 
to TDV when TDV is concomitantly administered with YF-17D vaccine compared to placebo. 
Safety parameters include solicited local (injection site) and solicited systemic adverse events 
(AE) for 7 days (day of vaccination + 6 days) and 14 days (day of vaccination + 13 days) after 
each vaccination, respectively, unsolicited AEs for 28 days (day of vaccination + 27 days) after 
each vaccination, medically attended adverse events (MAAE) and serious adverse events (SAE) 
throughout the trial. 
A schematic of the trial design is included as Figure 4.a. The schedule of trial procedures is 
provided in Appendix A. 
Immunogenicity evaluation: 
Blood samples for the measurement of dengue neutralizing antibodies (microneutralization test 
50% [MNT50]) will be collected at pre-first vaccination (Day 1 [M0]), 1-month post first 
vaccination (Day 30 [M1]), pre-second vaccination (Day 90 [M3]), 1-month post second 
vaccination (Day 120 [M4]), pre-third vaccination (Day 180 [M6]), and 1-month post third 
vaccination (Day 210 [M7]). 
Blood samples for the measurement of YF neutralizing antibodies (plaque reduction 
neutralization test [PRNT]) will be collected at pre-first vaccination (Day 1 [M0]), 1-month post 
first vaccination (Day 30 [M1]), pre-third vaccination (Day 180 [M6]), and 1-month post third 
vaccination (Day 210 [M7]). 
 
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Trial No. DEN-305 
Page 8 of 34
Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
Figure 4.a 
Schematic of Trial Design 
 
Safety evaluation: 
 Diary cards (paper or electronic) will be distributed to all subjects for the recording of: 
 Solicited local AEs for 7 days following each vaccination (day of vaccination + 6 days). 
These will include: injection site pain, injection site erythema, and injection site swelling, 
and will be collected at each injection site. 
 Solicited systemic AEs for 14 days following each vaccination (day of vaccination + 
13 days). These will include: fever, headache, asthenia, malaise, and myalgia. 
 Unsolicited AEs for 28 days following each vaccination (day of vaccination + 27 days). 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
 SAEs and MAAEs will be recorded for the trial duration. MAAEs are defined as AEs leading 
to an unscheduled visit to or by a healthcare professional including visits to an emergency 
department, but not fulfilling seriousness criteria. 
Data collection will be by electronic Case Report Form (CRF). 
 
 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
5.0 
ANALYSIS ENDPOINTS 
5.1 
Primary Endpoints 
 Proportion of subjects YF and DENV-naive at Baseline who are seroprotected against YF on 
Day 30 (Month-1 [M1]) as measured by PRNT (YF seroprotection rate). YF seroprotection is 
defined as reciprocal anti-YF neutralizing antibody titer ≥10. Immunological naivety to YF 
and DENV is defined as Baseline reciprocal neutralizing antibody titers <10 for YF and for 
the 4 dengue serotypes. 
5.2 
Secondary Endpoints 
Immunogenicity: 
 GMT of neutralizing antibodies (as measured by MNT50) for each of the 4 dengue serotypes 
at pre-second and -third vaccinations (Day 90 [Month 3; M3] and Day 180 [Month 6; M6], 
respectively); and 1 month post first, second, and third vaccinations (Day 30 [M1], Day 120 
[Month 4; M4], and Day 210 [Month 7; M7], respectively) in subjects YF and DENV-naive 
at Baseline. 
 GMTs of anti-YF neutralizing antibodies at 1-month post first and third vaccinations 
(Day 30 [M1] and Day 210 [M7], respectively) in subjects YF and DENV-naive at Baseline. 
 Seropositivity rates (% of subjects seropositive) for each of the 4 dengue serotypes at 
pre-second and -third vaccinations (Day 90 [M3] and Day 180 [M6], respectively); and 
1 month post first, second, and third vaccinations (Day 30 [M1], Day 120 [M4], and Day 210 
[M7], respectively) in subjects YF and DENV-naive at Baseline, where seropositivity is 
defined as a reciprocal neutralizing antibody titer ≥10. 
 Seropositivity rates (% of subjects seropositive) for multiple (2, 3 or 4) dengue serotypes at 
pre-second and -third vaccinations (Day 90 [M3] and Day 180 [M6], respectively); and 1 
month post first, second, and third vaccinations (Day 30 [M1], Day 120 [M4], and Day 210 
[M7], respectively) in subjects YF and DENV-naive at Baseline, where seropositivity is 
defined as a reciprocal neutralizing antibody titer ≥10. 
 Proportion of subjects YF and DENV-naive at Baseline who are seroprotected against YF at 
1-month post third vaccination (Day 210 [M7]) as measured by PRNT. 
Safety: 
 Frequency and severity of solicited local (injection site[s]) AEs for 7 days (day of 
vaccination + 6 days) and solicited systemic AEs for 14 days (day of vaccination + 13 days) 
after each vaccination. 
 Percentage of subjects with any unsolicited AEs within 28 days (day of vaccination + 
27 days) after each vaccination. 
 Percentage of subjects with MAAEs and SAEs throughout the trial. 
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Takeda’s Tetravalent Dengue Vaccine Candidate 
Trial No. DEN-305 
Page 11 of 34
Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
6.0 
DETERMINATION OF SAMPLE SIZE 
The sample size calculation assumes a significance level of 0.025 (one-sided). 
For the primary objective of showing NI in YF seroprotection rates, the calculation assumes a NI 
margin of 5%, and a YF seroprotection rate of 98% at 1 month after YF-17D vaccination in the 
two trial groups (Group 1 and Group 3). 
For the secondary objective of showing NI in GMTs of neutralizing antibodies for all the 4 
dengue serotypes, the calculation assumes a NI margin of 2.0, that the true GMT ratio for the two 
trial groups (Group 2 and Group 3) is 1, and that the natural logarithm of titers are distributed as 
normal distributions with SDs of 1.35, 0.86, 1.21, and 1.27. 
For the secondary objective of showing NI in GMTs of anti-YF neutralizing antibodies, the 
calculation assumes a NI margin of 2.0, that the true GMT ratio for the two trial groups (Group 1 
and Group 3) is 1, and that the natural logarithm of titers are distributed as normal distributions 
with a SD of 1.31 [1]. 
A sample size of 300 subjects per trial group, with approximately 255 evaluable subjects per trial 
group (assuming approximately 15% dropouts and non-evaluable subjects), is sufficient to 
achieve approximately 90% overall power for showing NI for the above primary and secondary 
objectives. 
A total sample size of 900 subjects also ensures that a sufficient number of healthy YF/DENV-
naive adults will be vaccinated to support the safe use of TDV in travelers. 
The power calculations were based on nQuery Advisor® 6.01. 
 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
7.0 
METHODS OF ANALYSIS AND PRESENTATION 
7.1 
General Principles 
This statistical analysis plan (SAP) was developed based on the information provided in Protocol 
DEN-305, Version 4.0 dated 28 November 2018 [2] and on International Conference on 
Harmonization (ICH) E3 [3] and E9 [4] Guidelines. 
All statistical analyses will be generated using the statistical analysis system SAS Version 9.2 or 
higher. 
A blinded data review will be conducted prior to unblinding of subject’s trial group assignment. 
This review will assess the accuracy and completeness of the trial database, subject evaluability, 
and appropriateness of the planned statistical methods. 
7.1.1 Data Presentation 
In general, descriptive summaries will be provided by trial group. 
Unless specified otherwise, number of subjects with non-missing observations, mean or 
geometric mean, SD or geometric SD, median, minimum, and maximum will be presented for 
continuous data. Frequency and percent will be presented for categorical data. In summary tables 
for categorical data for which categories are defined on the CRF, all categories will be presented 
as specified, even if the subject count within that category is zero. For other categorical data (eg, 
AEs and medications/vaccinations), only categories with at least one subject will be presented. 
Minimum and maximum values will be presented using the same number of decimal places as 
the recorded data. Means, geometric means, and medians will be presented to 1 more decimal 
place than the recorded data. SD and geometric SD will be presented to 2 more decimal places 
than the recorded data, with possible exceptions made for derived data. The CI about a parameter 
estimate will be presented using the same number of decimal places as the parameter estimate 
(ie, 1 more decimal place than the recorded data). Percentages will be presented to 1 decimal 
place (eg, 80.3%). 
All data collected will be presented in listings, sorted by trial group, site number, subject 
number, and date/time of the finding, if applicable. If not stated otherwise, screen failure subjects 
will be grouped and listed at the end. 
7.1.2 Study Day, Baseline and Analysis Window Definitions 
Study Day 1 is defined as the date of the first vaccination, as recorded in the CRF vaccination 
page. Other study days are defined relative to Study Day 1, with Day -1 being the day prior to 
Day 1. 
Baseline is defined as the last non-missing measurement taken before the first trial vaccination. 
Where time is available, the time of the measurement must be prior to the trial vaccination time. 
Day 1 observations taken after the vaccination are considered post-Baseline values. 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
A windowing convention for immunogenicity and safety (vital signs) data will be used to 
determine the analysis value of a variable at a given trial visit. Following the schedule of trial 
procedures (Appendix A), analysis windows for each visit will be calculated relative to the day 
on which each trial dose was administered (Day 1 [M0], Day 90 [M3], and Day 180 [M6]).  
The analysis visit windows for each trial visit are displayed in Table 7.a. 
Table 7.a 
Analysis Visit Windows 
Visit 
Scheduled 
Visit Day 
(Month)  
Scheduled 
Vaccination 
Analysis Visit Window 
Safety Set  
(Vital Signs) 
Full Analysis Set 
Per-Protocol Set 
V1 
Day 1 
(M0) 
Dose 1 
Prior [≤ 1 day] (a) to 
Dose 1 
Prior [≤ 1 day] (a) to 
Dose 1 
Prior [≤ 1 day] (a) to  
Dose 1 
V2 
Day 30 
(M1) 
2 – 60 (b) days after 
Dose 1 
2 – 60 (b) days after 
Dose 1 
29 – 37 (b) days after  
Dose 1 
V3 
Day 90 
(M3) 
Dose 2 
61 – 115 (b) days after 
Dose 1; and 
Prior [≤ 1 day] (a) to 
Dose 2 
61 – 115 (b) days after 
Dose 1; and 
Prior [≤ 1 day] (a) to 
Dose 2 
75 – 115 (b) after Dose 1; 
and  
Prior [≤ 1 day] (a) to  
Dose 2 
V4 
Day 120 
(M4) 
 
2 – 60 (b) days after 
Dose 2; or 
116 – 150 (b) days 
after Dose 1 (c) 
2 – 60 (b) days after 
Dose 2; or 
116 – 150 (b) days  
after Dose 1 (c) 
29 – 37 (b) days after  
Dose 2 
V5 
Day 180 
(M6) 
Dose 3 
61 – 115 (b) days after 
Dose 2; and 
Prior [≤ 1 day] (a) to 
Dose 3; or 
151 – 195 (b) days 
after Dose 1 (c) 
61 – 115 (b) days after 
Dose 2; and 
Prior [≤ 1 day] (a) to 
Dose 3; or 
151 – 195 (b) days  
after Dose 1 (c) 
75 – 115 (b) after Dose 2 
and  
Prior [≤ 1 day] (a) to  
Dose 3 
V6 
Day 210 
(M7) 
 
2 – 105 (b) days after 
Dose 3; or 
116 – 195 (b) days 
after Dose 2 (d); or 
196 – 285 (b) days 
after Dose 1 (c) 
2 – 105 (b) days after 
Dose 3; or 
≥116 (b) days after  
Dose 2 (d); or 
≥196 (b) days after  
Dose 1 (c) 
29 – 37 (b) days after  
Dose 3 
V7 
Day 360 
(M12) 
 
≥106 (b) days after 
Dose 3; or 
≥196 (b) days after  
Dose 2 (d); or 
≥286 (b) days after  
Dose 1 (c) 
Not applicable (no 
blood sample is taken 
at V7) 
Not applicable (no  
blood sample is taken  
at V7) 
(a) Blood draw for immunogenicity assessments and assessments of vital signs must be prior to the vaccination scheduled for 
the same visit, and where time is available, the time of the blood/vital signs collection must be prior to the vaccination time. 
Day 1 (M0) observations taken after the first trial vaccination are considered post-Baseline values. 
(b) Number of days after the visit is calculated with 1 day increment. For example, for V2 number of days after V1 is calculated 
as [Date of V2] – [Date of V1] + 1 (day). 
(c) Applies to subjects who missed Dose 2, or both Dose 2 and Dose 3. 
(d) Applies to subjects who missed Dose 3. 
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Statistical Analysis Plan (Final, Version 1.0) 
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If several measurements of a variable are obtained for a subject within the same visit window, 
the measurement with the date closest to the scheduled visit date will be used. In the event that 2 
measurements within a given visit window are equidistant to the scheduled visit date, the later 
observation will be used. Both scheduled and unscheduled visits will be considered equally. 
7.1.3 Handling of Missing Data 
Data will be presented in the listings as reported. For the summaries and analysis, the following 
conventions apply. 
Immunogenicity data 
Dengue neutralizing antibody titers (MNT50) which are below the lower limit of detection 
(LLOD, 10) will be imputed with a value of 5 (half of the LLOD). If a reported value is between 
the LLOD and the lower limit of quantification (LLOQ, which differs among serotypes), this 
value will be imputed with the mid-point between the LLOD and LLOQ. For example, given a 
LLOQ of 18 for a serotype, all values between 10 and 18 will be imputed with 14 for this 
serotype. 
YF antibody titers (PRNT) which are below the LLOQ (ie, <10) will be imputed with a value of 
5 (half of the LLOQ). 
No imputation method will be used for missing immunogenicity data and analyses will be based 
on complete records only. 
Adverse event data 
Missing information regarding ‘relationship to investigational product (IP)’ (related/not related) 
for solicited systemic and unsolicited AEs, and ‘severity’ (mild/moderate/severe) for unsolicited 
AEs, will be handled using the worst-case approach. Thus, unsolicited AEs with missing severity 
will be considered as ‘severe’ and solicited systemic and unsolicited AEs with missing 
relationship will be considered as ‘related’. 
Missing and partial unsolicited AE start dates may be imputed only to determine the temporal 
relationship between the start date of the event and the dose date of the most appropriate 
vaccination that the AE should be allocated with (i.e. Vaccination 1, 2, or 3). An AE should be 
temporally allocated with the correct dose using the following rules: 
 If the AE start and end dates are both completely missing, the AE will be allocated with 
the first trial vaccination; 
 If at least month and/or year of AE start is/are available, the AE will be allocated with the 
latest vaccination prior to the AE start date; 
 If the AE start date is completely missing, or the available start date information is 
insufficient to distinguish between the 3 trial vaccinations, but an AE end date or a partial 
AE end date (ie, month and/or year) is available, the AE end date will be assessed, and 
the AE will be allocated with the vaccination after which the event ends. This is based on 
the assumption that any AE starting after Vaccination 1 or 2 and ongoing on the day of 
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Statistical Analysis Plan (Final, Version 1.0) 
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Vaccination 2 or 3 would be identified during the clinical assessments that are performed 
before administration of the second or third trial vaccination. If partial end date 
information indicates possible association with multiple vaccinations, the AE will be 
allocated with the earliest of these. 
Prior/concomitant medication/vaccination data 
Missing and partial medication/vaccination dates may be assessed only to distinguish between a 
prior or a concomitant medication/vaccine. A medication will be considered prior only if the 
partial end date indicates that it was stopped before first trial vaccination. A vaccine will be 
considered prior only if the partial vaccination date indicates that it was given before the first 
trial vaccination. In all other cases the medication or vaccine will be considered concomitant. 
Medical history/concurrent medical conditions 
In case the “End Date” or “End Date Unknown” fields are missing on the medical history/ 
concurrent medical conditions form of the CRF and from the partial date it can’t be concluded 
that the event is clearly a medical history, the event will be considered concurrent medical 
condition. 
7.1.4 Implausible Values 
Data outside the plausible ranges as defined in Table 7.b will be excluded from respective 
analyses, but presented as recorded in data listings including a flag that highlights implausible 
values. 
Table 7.b 
Plausible Data Ranges 
 
Parameter 
Plausible range 
Demographic/ 
Physical examination 
Height 
110 – 210 cm 
Weight 
20 – 200 kg 
Solicited AEs 
Erythema 
≤ 500 mm 
Swelling 
≤ 500 mm 
Body Temperature (a) 
32 – 43ºC 
Vital Signs 
Heart Rate 
40 – 200 beats/min 
Systolic Blood Pressure 
70 – 180 mmHg 
Diastolic Blood Pressure 
30 – 120 mmHg 
Respiratory Rate 
5 – 80 breaths/min 
(a) Also applicable to body temperature measurements collected as vital signs. 
7.2 
Analysis Sets 
All Screened: All subjects who signed the informed consent, regardless of whether subjects 
were screen failures. 
Randomized Set: All randomized subjects, regardless of whether any dose was received. 
Safety Set: All randomized subjects who received at least 1 dose of IP. 
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Full Analysis Set (FAS): All randomized subjects who received at least 1 dose of IP and for 
whom a valid pre-dose (Baseline) and at least 1 post-dose measurement are available for 
immunogenicity assessments. 
YF FAS: All randomized subjects who received at least 1 dose of IP and for whom a valid pre-
dose (Baseline) and at least 1 post-dose measurement (Day 30) are available for YF 
immunogenicity assessments. 
Per-Protocol Set (PPS): All subjects from the FAS who have no major protocol violations, 
excluding subjects who are seroprotected for YF virus and/or who are seropositive for any 
serotype of dengue virus at Baseline. 
YF PPS: All subjects from the YF FAS who have no major protocol violations, excluding 
subjects who are seroprotected for YF virus and/or who are seropositive for any serotype of 
dengue virus at Baseline. 
The criteria described in Table 7.c will be used to identify subjects who will be excluded from 
the PPS and YF PPS during the blinded data review prior to unblinding of the IP assignment. 
These criteria are considered to have a potentially significant impact on the immunogenicity 
results of the subject. Subjects excluded from PPS and YF PPS due to receiving incorrect IP will 
be identified after unblinding. 
Other major protocol violations may be identified during the data review and deviation logs 
throughout the trial, subject to medical review. Any changes to these criteria after approval of the 
SAP will be documented and approved prior to database lock. 
Analyses using the Randomized Set, FAS sets, and PPS sets will be based on randomized 
treatment. 
Analyses using the Safety Set will be based on the actual treatment. For example, a subject 
randomized to Group 1 (YF+P/TDV/TDV) but vaccinated with the planned vaccinations of 
Group 2 (TDV + P/TDV/YF) will be analyzed in Group 2 (TDV + P/TDV/YF). Subjects who 
received vaccination (or combination of vaccinations) that is not planned for any trial group will 
be considered together in a separate group. Data for this group will be displayed in selected 
summaries and all listings generated for the Safety Set. 
Analyses based on the Safety Set (except AEs), FAS and PPS sets will only include 
measurements obtained following the analysis visit windows defined in Table 7.a. 
 
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Statistical Analysis Plan (Final, Version 1.0) 
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Table 7.c 
Criteria for Exclusion from the PPS 
Criteria for Exclusion 
Probable Method of Identification 
YF PPS 
PPS 
Not receiving at least one dose of the IP 
(a) 
Identified programmatically using 
dosing data 
X 
X 
Not having a valid pre-dose (Baseline) 
and at least 1 valid post-dose 
measurement for YF immunogenicity 
assessments at Day 30 (b) 
Identified programmatically using 
immunogenicity data 
X  
NA 
Not having a valid pre-dose (Baseline) 
and at least 1 valid post-dose 
measurement for immunogenicity 
assessments (b) 
Identified programmatically using 
immunogenicity data 
NA  
X 
Subjects seroprotected for YF virus 
and/or seropositive to any serotype of 
dengue neutralizing antibody titers at 
Baseline (Day 1 [M0]) 
Identified programmatically using 
immunogenicity data 
X 
X 
Not receiving all three doses of the IP 
Identified programmatically using 
dosing data 
NA 
X 
Receiving the second trial and/or third 
vaccination inadmissibly outside of the 
scheduled visit window (ie, outside Day 
90 [-15/+25 days]) 
Identified programmatically using 
dosing data 
NA 
X 
Receiving incorrect IP(s) for Vaccination 
1, Vaccination 2, and/or Vaccination 3 
Identified after unblinding (eg, a subject 
who was randomized to TDV but 
received YF or a subject who was 
randomized to Placebo but received 
TDV) 
X (c) 
X 
Receiving IP using incorrect route of 
administration  
Identified through protocol deviation 
log 
X (c) 
X 
Product preparation error 
Identified through protocol deviation 
log 
X 
X 
Subject meets any of the following 
exclusion criteria: 5-8, 19-23 
Identified programmatically using CRF-
recorded data; Identified through 
protocol deviation review/medical 
review 
X 
X 
Use of prohibited medications/vaccines 
Identified through medical review based 
on CRF-recorded data 
X (c) 
X 
(a) Subjects with this protocol violation will be excluded from the Safety Set, and thus also from the FAS sets and 
the PPS sets. 
(b) Subjects with this protocol violation will be excluded from the YF FAS/FAS, and thus also from the PPS sets. 
(c) Applies to Vaccination 1 only. 
7.3 
Disposition of Subjects 
Trial information will be presented for all screened subjects, including date first subject signed 
the informed consent form, the date of first subject’s first vaccination, the date of last subject’s 
first vaccination, the date of first subject’s second vaccination, the date of last subject’s second 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
vaccination, the date of first subject’s third vaccination, the date of last subject’s third 
vaccination, the date of the first subject’s first visit, the date of the last subject’s last visit, the 
Medical Dictionary for Regulatory Activities (MedDRA) version, the World Health 
Organization Drug Dictionary (WHO-Drug) version, and the SAS version used for analysis. 
Disposition of all screened subjects will be summarized descriptively, including a summary of 
the number of screened subjects, the number of randomized subjects and the primary reason for 
not being eligible for randomization. The number of screen failures and their characteristics will 
also be summarized. 
Disposition for all randomized subjects will be summarized by trial group. Disposition categories 
include: 
 Number of subjects randomized by trial site. 
 Number of subjects randomized, but not vaccinated (including primary reason for being 
randomized, but not vaccinated). 
 Number of subjects who completed the vaccine regimen/trial visits. 
 Number of subjects who prematurely discontinued the vaccine regimen/trial visits. 
 Primary reason for premature discontinuation of the vaccine regimen/trial visits. 
Number of subjects in the analysis sets will be summarized by trial group. Reasons for exclusion 
of subjects from the analysis sets will be summarized by trial group based on all randomized 
subjects. In addition, significant protocol deviations will be summarized by trial group for the 
Randomized Set. 
7.4 
Demographic and Other Baseline Characteristics 
Age, gender, race, and other Baseline characteristics will be summarized descriptively based on 
the Safety Set, YF FAS, FAS, YF PPS, and PPS. Baseline characteristics for the FAS sets will 
also include baseline seropositivity status for dengue (seropositive for at least 1 dengue serotype 
/seronegative for all dengue serotypes), baseline seropositivity status for each and multiple 
dengue serotype, and baseline seroprotection status against YF (yes/no). 
7.5 
Medical History and Concurrent Medical Conditions 
Medical history and concurrent medical conditions will be coded according to the MedDRA 
coding system. 
A medical history is defined as any significant condition/disease that stopped at or prior to first 
dose of IP. A concurrent medical condition is defined as any significant condition/disease that is 
ongoing at the time the first dose of IP is administered. 
Summary tables will be provided by SOC and PT based on the Safety Set. 
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Statistical Analysis Plan (Final, Version 1.0) 
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7.6 
Medication History and Concomitant Medications 
Medication history, vaccination history, concomitant medications, and concomitant vaccinations 
will be coded according to WHO Drug. 
A prior medication/vaccination (history) is any medication/vaccination which intake was stopped 
before first dose of IP. A concomitant medication/vaccination is any medication/ vaccination 
ongoing at the time the first dose of IP is administered, or taken/administered on/after the first 
dose of IP. 
Summary tables for medication history and concomitant medications will be provided by 
Anatomical Therapeutic Chemical class level 2 and preferred medication name. Vaccination 
history and concomitant vaccination will be summarized by vaccination type and name as 
recorded in the CRF. Summaries and listings will be provided based on the Safety Set. 
7.7 
Investigational Product Exposure and Compliance 
The Investigator records all injections of the IP given to the subject on the CRF. 
Investigational product compliance will be summarized for the Safety Set presenting the number 
and percentage of subjects receiving: 
1) First vaccination only; 
2) First and second vaccinations only; 
3) All three vaccinations. 
This summary will be prepared by trial group, including a separate group of subjects who 
received different IPs (or combination of different IPs) that is not planned for any trial group (if 
any). 
The duration of follow-up [days] after each dose will be summarized for the Safety Set as a 
continuous variable (number of subjects [n], mean, SD, median, minimum, and maximum), and 
as categorical variable (frequency and percentage of subjects) for the following categories: 
After first vaccination: 
 1 – 30 days, 
 31 – 90 days, 
 91 – 120 days, 
 121 – 180 days, 
 181 – 360 days, 
 ≥ 361 days.  
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Statistical Analysis Plan (Final, Version 1.0) 
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After second vaccination: 
 1 – 30 days, 
 31 – 90 days, 
 91 – 120 days, 
 121 – 180 days, 
 181 – 270 days, 
 ≥ 271 days. 
After third vaccination: 
 1 – 30 days, 
 31 – 90 days, 
 91 – 120 days, 
 121 – 180 days, 
 ≥ 181 days. 
The duration of follow-up will be calculated as end of trial date – vaccination date (first, second 
or third, respectively) + 1 day. 
7.8 
Efficacy Analysis 
Not applicable. 
7.9 
Pharmacokinetic/Pharmacodynamic Analysis 
Not applicable. 
7.10 
Other Outcomes 
The primary and secondary immunogenicity endpoints are listed in Table 7.d, together with the 
analyses (descriptive summaries, graphical presentations, or pairwise comparisons) planned for 
each endpoint/population. The primary population for all immunogenicity summaries and analyses 
will be the YF-PPS/PPS. Supportive immunogenicity summaries will be provided based on the 
YF-FAS/FAS. 
Descriptive Summaries 
Descriptive statistics for the primary and secondary immunogenicity endpoints will be provided 
by trial group and for each vaccine/serotype, and will include the following: 
 Seroprotection/Seropositivity Rate: number and percentage of subjects with 
seroprotection / seropositivity and corresponding 95% CIs calculated with the exact 
(Clopper-Pearson) method [5]. 
 GMT: geometric mean, GSD, median, 95% CIs, minimum, and maximum. The 
geometric mean, GSD, and the 95% CIs of the geometric mean will be calculated as the 
anti-logarithm transformation of the mean, standard deviation, and 95% CIs of the log-
transformed titers. 
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Statistical Analysis Plan (Final, Version 1.0) 
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Graphical Presentations 
Graphical presentations for selected immunogenicity endpoints will be provided by trial group 
and for each vaccine/serotype, and will include the following: 
 Bar graphs presenting the percentage of subjects with seroprotected (YF)/seropositive 
(for each of the 4 dengue serotypes and for multiple serotypes [at least trivalent and 
tetravalent only], including error bars for the 95% CIs, for all visits (except baseline). 
 Line plots of GMTs, including error bars for the 95% CIs, plotted over time. 
 Reverse cumulative distribution curves of antibody titers 
o For YF at Day 30 (Group 1 and Group 3) and Day 210 (Group 2); 
o For Dengue at Day 120 (Group 2 and Group 3) and Day 210 (Group 1). 
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Statistical Analysis Plan (Final, Version 1.0) 
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Table 7.d 
Planned Analyses and Populations for Immunogenicity Endpoints 
Endpoint 
Planned Analyses and Populations 
Vaccine 
Variable 
Time 
Point 
Endpoint 
Type 
Descriptive 
Summaries 
Graphical 
Presentations 
Pairwise 
Comparisons(a) 
YF 
Seroprotection 
Rate 
Day 30 
Primary 
YF PPS,  
YF FAS 
 YF PPS 
YF PPS,  
YF FAS 
Day 180 
Secondary 
PPS, FAS 
 PPS 
Day 210 
GMT 
Day 30 
Secondary 
YF PPS,  
YF FAS 
YF PPS 
YF PPS,  
YF FAS 
Day 180 
Secondary 
PPS, FAS 
PPS 
 
Day 210 
Dengue 
Seropositivity 
Rate  
(Each serotype(a)) 
Day 30 
Secondary 
PPS, FAS 
PPS 
Day 90 
Day 120 
Day 180 
Day 210 
GMT  
(Each serotype(b)) 
Day 30 
Secondary 
PPS, FAS 
PPS 
PPS, FAS 
(Day 120) 
Day 90 
Day 120 
Day 180 
Day 210 
Seropositivity 
Rate  
(Multiple 
serotypes(c)) 
Day 30 
Secondary 
PPS, FAS 
PPS 
Day 90 
Day 120 
Day 180 
Day 210 
(a) Pre-specified comparisons of interest are defined in the following text. 
(b) Immunogenicity for each dengue serotype will be summarized /analyzed for: DENV-1, DENV-2, DENV-3, and DENV-4. 
(c) Immunogenicity for multiple dengue serotypes will be summarized in the following categories: Monovalent, Bivalent, 
Trivalent, Tetravalent, at least Bivalent, and at least Trivalent. 
Pairwise Comparisons 
Pairwise comparison will be performed for the primary endpoint and key secondary endpoints 
(with the later group as the reference group): 
 YF Seroprotection Rate: Group 1 vs Group 3 at Day 30. 
 YF GMT: Group 1 vs Group 3 at Day 30. 
 TDV GMT: Group 2 vs Group 3 at Day 120. 
The following statistics will be provided for the comparison: 
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Statistical Analysis Plan (Final, Version 1.0) 
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 Seroprotection Rate: rate difference and corresponding 95% CIs based on the Newcombe 
score method [6]; risk ratio and corresponding 95% CIs based on Fisher’s exact test. 
 GMT: geometric mean ratio and 95% CIs based on an analysis of variance (ANOVA) 
model, which includes the log-transformed value of titer as the dependent variable, and 
the trial group as a factor. The geometric mean, geometric mean ratio and 95% CIs are 
presented in anti-log values of least squares means (LS means) estimated from the 
ANOVA model. 
7.10.1 Primary Immunogenicity Endpoint 
The primary endpoint of this trial is the proportion of subjects (YF and DENV-naive at Baseline) 
who are seroprotected against YF on Day 30 (M1) as measured by PRNT (YF seroprotection 
rate). YF seroprotection is defined as reciprocal anti-YF neutralizing antibody titer ≥10. 
Immunological naivety to YF and DENV is defined as Baseline reciprocal neutralizing antibody 
titers <10 for YF and for the 4 dengue serotypes. 
Descriptive summary and graphical presentation will be provided for the primary 
immunogenicity endpoint based on the YF PPS. Sensitivity analyses will be provided based on 
the YF FAS. The primary pairwise comparison, i.e. Group 1 vs Group 3 in terms of YF 
seroprotection rates on Day 30, will be carried out based on the YF PPS, with a sensitivity 
analysis performed based on the YF FAS. 
The NI of the immune response to YF-17D vaccine when concomitantly administered with TDV 
(Group 3) compared to concomitant administration of YF-17D vaccine with placebo (Group 1) 
will be assessed in terms of seroprotection rates on Day 30 (M1) based on the YF PPS. NI will 
be concluded if the upper bound of the 95% CI for the seroprotection rate difference (Group 1 
minus Group 3) is less than the NI margin of 5%. 
Sensitivity analyses for the primary endpoint based on the YF FAS will include all subjects 
regardless of their DENV/YF protection status at Baseline. 
7.10.2 Secondary Immunogenicity Endpoints 
The secondary immunogenicity endpoints of this trial are: 
 GMT of neutralizing antibodies (as measured by MNT50) for each of the 4 dengue serotypes 
at pre-second and -third vaccinations (Day 90 [Month 3; M3] and Day 180 [Month 6; M6], 
respectively); and 1 month post first, second, and third vaccinations (Day 30 [M1], Day 120 
[Month 4; M4], and Day 210 [Month 7; M7], respectively) in subjects YF and DENV-naive 
at Baseline. 
 GMTs of anti-YF neutralizing antibodies at 1-month post first and third vaccinations (Day 
30 [M1] and Day 210 [M7], respectively) in subjects YF and DENV-naive at Baseline. 
 Seropositivity rates (% of subjects seropositive) for each of the 4 dengue serotypes at pre-
second and -third vaccinations (Day 90 [M3] and Day 180 [M6], respectively); and 1 month 
post first, second, and third vaccinations (Day 30 [M1], Day 120 [M4], and Day 210 [M7], 
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respectively) in subjects YF and DENV-naive at Baseline, where seropositivity is defined as 
a reciprocal neutralizing antibody titer ≥10. 
 Seropositivity rates (% of subjects seropositive) for multiple (2, 3 or 4) dengue serotypes at 
pre-second and -third vaccinations (Day 90 [M3] and Day 180 [M6], respectively); and 1 
month post first, second, and third vaccinations (Day 30 [M1], Day 120 [M4], and Day 210 
[M7], respectively) in subjects YF and DENV-naive at Baseline, where seropositivity is 
defined as a reciprocal neutralizing antibody titer ≥10. 
 Proportion of subjects YF and DENV-naive at Baseline who are seroprotected against YF at 
1-month post third vaccination (Day 210 [M7]) as measured by PRNT. 
As specified in Table 7.d, descriptive summaries, graphical presentations, and pairwise 
comparisons will be based on the YF PPS and PPS. Sensitivity analyses will be based on the YF 
FAS and FAS, respectively. 
The NI of the immune response to TDV when concomitantly administered with YF-17D vaccine 
(Group 3) compared to concomitant administration of TDV with placebo (Group 2) will be 
assessed in terms of the GMTs of neutralizing antibodies for all 4 dengue serotypes on Day 120 
(M4). NI will be concluded if the upper bound of the 95% CI for the GMT ratio (Group 2/Group 
3) is less than the NI margin of 2.0. An ANOVA model will be used for this assessment. 
The NI of the immune response to YF-17D vaccine when concomitantly administered with TDV 
(Group 3) compared to concomitant administration of YF-17D with placebo (Group 1) will be 
assessed in terms of the GMTs of anti-YF neutralizing antibodies on Day 30 (M1). NI will be 
concluded if the upper bound of the 95% CI for the GMT ratio (Group 1/Group 3) is less than the 
NI margin of 2.0. An ANOVA model will be used for this assessment. 
Sensitivity analyses for secondary endpoints based on the YF FAS or FAS will include all 
subjects regardless of their DENV/YF protection status at Baseline. 
7.11 
Safety Analysis 
All summaries and analyses of safety data will be based on subjects in the Safety Set. 
7.11.1 Adverse Events 
Unless otherwise specified, AEs will be summarized by trial group after first trial vaccination, 
second trial vaccination, third trial vaccination, and any trial vaccination. 
Reactogenicity (Solicited AEs) 
Solicited AEs are collected for at least 30 min after each vaccination at the site (in-clinic 
assessment). In addition, subjects are provided with a diary card for the recording of solicited 
local (injection site) AEs, including injection site pain, injection site erythema, and injection site 
swelling, for 7 days following vaccination (day of vaccination + 6 days). Subjects are also 
provided with a diary card for the recording of solicited systemic AEs (fever, headache, asthenia, 
malaise, and myalgia) for 14 days following vaccination (day of vaccination + 13 days). For the 
local (injection site) AEs erythema and swelling, the subject/the subject’s representative will 
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Statistical Analysis Plan (Final, Version 1.0) 
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record the length of the longest diameter in mm. However, for the analysis these data will be 
displayed in cm. For the systemic AE fever, the subject/the subject’s representative will record 
the body temperature in either °F or °C. For the analysis, all data will be displayed in °C. 
Severity grades for erythema and swelling will be derived from the recorded diameters, and fever 
will be presented using the proposed temperature increments published by the Brighton 
Collaboration Fever Working Group [7]. Details of solicited local (injection site) and systemic 
AEs, and severity of solicited safety parameters are given in Appendix B. 
Missing data for solicited AEs will not be imputed unless otherwise specified in Section 7.1.3. 
For each trial group and solicited AE, the denominator for the percentage will exclude subjects 
with completely missing data (i.e. subject does not have at least 1 recorded results [ie, none, 
mild, moderate, or severe]) for the solicited AE in the period being summarized. 
For each solicited AE, the number and percentage of subjects reporting an event will be 
summarized by event severity at the following intervals post-vaccination: 
 30 minutes (in-clinic assessment of solicited local [injection site] and systemic AEs – 
analyzed separately from diary-recorded solicited AEs); 
 Within 7 days (solicited local [injection site] AEs); 
 Within 14 days (solicited systemic AEs); 
 Days 1 – 7 (daily, solicited local [injection site] AEs); 
 Days 1 – 14 (daily, solicited systemic AEs); 
 Days 1 – 3 and Days 4 – 7 (solicited local [injection site] AEs); 
 Days 1 – 7 and Days 8 – 14 (solicited systemic AEs). 
For subjects with more than 1 episode of the same event, the maximum severity will be used for 
tabulations. 
Concomitantly administered vaccines (e.g., YF + Placebo) will be injected into opposite arms at 
Vaccination 1. Solicited local events following Vaccination 1 will be summarized for the two 
arms respectively, and presented as YF Arm, TDV Arm, or Placebo Arm, by linking left/right 
arm with kit numbers received after unblinding. Summaries of these solicited local events will be 
tabulated by each arm and for any arm as describe in Table 7.e. 
Table 7.e 
Summaries of Solicited Local AEs following Vaccination 1 
 
Vaccination 1 
Summary #1 
Arm (1) 
Summary #2 
Arm (2) 
Summary #3 
Any Arm 
Group 1 
YF + Placebo 
YF arm 
Placebo arm 
Any 
Group 2 
TDV + Placebo 
Placebo arm 
TDV arm 
Any 
Group 3 
TDV + YF 
YF arm 
TDV arm 
Any 
 
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For solicited systemic AEs, the number and percentage of subjects will also be summarized by 
relationship to IP (assessed by the Investigator) for the following intervals post-vaccination: 
 30 minutes; 
 Within 14 days. 
Subjects will only be counted once if the subject has more than 1 episode of the same event. In 
the case where the subject has both related and unrelated solicited systemic AEs, the subject will 
be counted under the related category. All solicited local (injection site) AEs are considered as 
related to IP. 
A summary of the day of first onset of each event and the number of days subjects reported each 
event will be presented post-vaccination. The number of days a subject reported each event is 
calculated as the total of all days the subject reported the event, regardless of whether the event 
was reported on consecutive days. 
Prolonged solicited AEs that continued beyond Day 7 (for local [injection site] AEs) or Day 14 
(for systemic AEs) will be captured in the AE CRF page, indicated by the checkbox “Is this 
event a continuation of a solicited event”. These prolonged solicited AEs will be presented in a 
separate listing and will not be included in any unsolicited AE summary or listing. 
An overview table for solicited AEs post-vaccination will be provided including: 
 30 minutes in-clinic assessment (solicited local [injection site] and systemic AEs 
combined). 
 Solicited AEs (solicited local [injection site] and systemic AEs combined). 
 Solicited local [injection site] AEs. 
 Solicited systemic AEs (overall and by relationship to IP). 
 Prolonged solicited AEs (overall and for solicited local [injection site] and systemic AEs 
separately). 
Unsolicited AEs 
Unsolicited AEs will be assessed for 28 days following administration of each trial dose (day of 
vaccination + 27 days). MAAEs, SAEs and AEs leading to IP withdrawal and/or trial 
discontinuation will be collected from first trial dose until the end of the trial. Unsolicited AEs, 
MAAEs, SAEs, and AEs leading to IP withdrawal and/or trial discontinuation will be coded 
according to the current version of MedDRA. 
In general, the number of events, number of subjects, and the percentage of subjects will be 
tabulated by trial group at each of the following levels: overall (any AEs/subjects with any AEs) 
and by SOC and PT. Subjects reporting more than 1 occurrence for the term (level) being 
summarized will be counted only once in number/percentage of subjects. Percentages will be 
based on the number of subjects in the Safety Set who received the respective trial dose. When 
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relationship or severity is concerned, the AE with the most closely related occurrence or the 
highest known severity will be counted. 
Unsolicited AEs up to 28 days post-vaccination will be summarized as follows: 
 By SOC and PT. This summary will also include a separate group of subjects who 
received different IPs (or combination of different IPs) that is not planned for any trial 
group (if any); 
 By SOC and PT including events with frequency greater than 2% in any trial group; 
 By SOC and PT for IP related events; 
 By SOC and PT for IP related events with frequency greater than 2% in any trial 
group; 
 By SOC, PT, and severity (mild, moderate, severe). 
MAAEs post-vaccination throughout the trial duration will be summarized as follows: 
 By SOC and PT; 
 By SOC and PT for IP related events; 
 By SOC, PT, and severity (mild, moderate, severe). 
SAEs post-vaccination throughout the trial duration will be summarized as follows: 
 By SOC and PT. This summary will also include a separate group of subjects who 
received different IPs (or combination of different IPs) that is not planned for any trial 
group (if any). 
 By SOC and PT for IP related events. 
AEs leading to IP withdrawal and/or trial discontinuation post-vaccination throughout the trial 
duration will be summarized as follows: 
 By SOC and PT. This summary will also include a separate group of subjects who 
received different IPs (or combination of different IPs) that is not planned for any trial 
group (if any). 
 By SOC and PT for IP related events. 
Subject mappings (ie, list of subject numbers in each category of SOC and PT, and each trial 
group) will be provided for unsolicited AEs, SAEs, MAAEs and AEs leading to IP withdrawal 
and/or trial discontinuation. 
Based on clinicaltrials.gov results posting requirements another summary table by SOC and PT 
for AEs after any vaccination is needed including all non-serious events (ie, all non-serious AEs 
up to 28 days post-vaccination, all MAAEs post-vaccination throughout the trial duration, and all 
non-serious AEs leading to IP withdrawal and/or trial discontinuation post-vaccination 
throughout the trial duration) with frequency greater than 2% in any trial group. 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
In addition, overview tables by trial group will be generated for unsolicited AEs (collected up to 
28 days post-vaccination), SAEs, MAAEs, and AEs leading to IP withdrawal and/or trial 
discontinuation including the variables as outlined in Table 7.f. 
Table 7.f 
Overview of Unsolicited Adverse Events 
 
All AEs 
(28 days post-
vaccination) 
SAEs 
MAAEs 
AEs leading to  
IP withdrawal 
and/or trial 
discontinuation 
Relationship to IP 
 
 
 
 
Relationship to trial procedure 
 
 
 
 
Severity 
 
 
 
 
AEs leading to IP withdrawal and/or 
trial discontinuation 
 
 
 
 
AEs leading to IP withdrawal 
 
 
 
 
AEs leading to trial discontinuation 
 
 
 
 
MAAEs 
 
 
 
 
SAEs and Non-serious AEs 
 
 
 
 
Deaths 
 
 
 
 
7.11.2 Clinical Laboratory Evaluations 
Not applicable. 
7.11.3 Vital Signs 
Vital sign will be summarized descriptively (number of subjects [n], mean, SD, median, 
minimum, and maximum) at each scheduled visit. The change from Baseline to each scheduled 
post-Baseline visit will be presented as applicable. 
7.11.4 12-Lead ECGs 
Not applicable. 
7.11.5 Other Observations Related to Safety 
Not applicable. 
7.12 
Changes in the Statistical Analysis Plan 
 A separate set of FAS and PPS was defined for the endpoints related to YF at Day 30. 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
8.0 
REFERENCES 
1. Nasveld PE, Marjason J, Bennett S, Aaskov J, Elliott S, McCarthy K, et al. Concomitant or 
sequential administration of live attenuated Japanese encephalitis chimeric virus vaccine and 
yellow fever 17D vaccine: randomized double-blind phase II evaluation of safety and 
immunogenicity. Hum Vaccin 2010;6(11):906-14. 
2. A Randomized, Observer-Blind, Placebo-Controlled, Phase 3 Trial to Investigate the 
Immunogenicity and Safety of a Tetravalent Dengue Vaccine Candidate and a Yellow Fever 
YF-17D Vaccine Administered Concomitantly and Sequentially in Healthy Subjects Aged 
18 to 60 years in Non-Endemic Country(ies), Takeda Vaccines, Inc., Protocol No. DEN-305, 
Version 4.0, dated 28 November 2018. 
3. ICH Harmonized Tripartite Guideline – Clinical Trial Reports: Structure and Content, E3 
(http://www.ich.org/products/guidelines/efficacy/efficacy-single/article/structure-and-
content-of-clinical-study-reports.html). 
4. ICH Harmonized Tripartite Guideline – Statistical Principles for Clinical Trials, E9 
(http://www.ich.org/products/guidelines/efficacy/efficacy-single/article/statistical-principles-
for-clinical-trials.html). 
5. Clopper CJ and Pearson ES. The use of confidence or fiducial limits illustrated in the case of 
the binomial. Biometrika. 1934;26:404-13. 
6. Newcombe RG. Two‐sided confidence intervals for the single proportion: comparison of 
seven methods. Statistics in Medicine. 1998;17.8:857-72. 
7. Marcy SM, Kohl KS, Dagan R, Nalin D, Blum M, Jones MC, et al. Fever as an adverse 
event following immunization: case definition and guidelines of data collection, analysis and 
presentation. Vaccine. 2004;22(5-6):551-6. 
 
 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
CONFIDENTIAL 
Appendix A Schedule of Trial Procedures 
Table 8.a 
Schedule of Trial Procedures 
Visit number 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
Day/Month 
Day 1 
 
M0 
Day 30 
 
M1 
Day 90 
 
M3 
Day 120 
(V3+30 days) 
M4 
Day 180 
(V3+90 days) 
M6 
Day 210 
(V5+30 days) 
M7 
Day 360 
(V5+180 days) 
M12 (ET) (a) 
Visit window (days) 
±0 
-1/+7 
-4/+7 
-1/+7 
-4/+7 
-1/+7 
-7/+14 
Signed informed consent (b) 
X 
 
 
 
 
 
 
Assessment of eligibility criteria (c) 
X 
 
 
 
 
 
 
Demographics (b) 
X 
 
 
 
 
 
 
Medical history (b) 
X 
 
 
 
 
 
 
Concomitant medications/ 
vaccinations (d) 
X 
X 
X 
X 
X 
X 
X 
Check contraindications to vaccination 
 
 
X 
 
X 
 
 
Check criteria for delay of vaccination 
 
 
X 
 
X 
 
 
Complete physical examination (e) 
X 
 
X 
 
X 
 
 
Targeted physical examination (f) 
 
X 
 
X 
 
X 
X 
Vital signs (g) 
X 
X 
X 
X 
X 
X 
X 
Pregnancy test (h) 
X 
 
X 
 
X 
 
 
Pregnancy avoidance guidance (i) 
X 
X 
X 
X 
X 
X 
 
Randomization 
X 
 
 
 
 
 
 
Blood Collection 
YF neutralizing 
antibodies (10 mL) (j) 
X 
X 
 
 
X 
X 
 
Dengue neutralizing 
antibodies (5 mL) (j) 
X 
X 
X 
X 
X 
X 
 
Vaccine administration 
X 
 
X 
 
X 
 
 
30 min post-vaccination in-clinic observation 
X 
 
X 
 
X 
 
 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
CONFIDENTIAL 
Visit number 
V1 
V2 
V3 
V4 
V5 
V6 
V7 
Day/Month 
Day 1 
 
M0 
Day 30 
 
M1 
Day 90 
 
M3 
Day 120 
(V3+30 days) 
M4 
Day 180 
(V3+90 days) 
M6 
Day 210 
(V5+30 days) 
M7 
Day 360 
(V5+180 days) 
M12 (ET) (a) 
Visit window (days) 
±0 
-1/+7 
-4/+7 
-1/+7 
-4/+7 
-1/+7 
-7/+14 
Including injection site evaluation and body 
temperature measurement (k) 
Diary Card (l) 
Distribution 
X 
 
X 
 
X 
 
 
Review/collection 
 
X 
 
X 
 
X 
 
Unsolicited AEs (m) 
X 
X 
X 
X 
X 
X 
 
SAEs (n) 
X 
X 
X 
X 
X 
X 
X 
MAAEs (n) 
X 
X 
X 
X 
X 
X 
X 
AEs leading to discontinuation or withdrawal 
(n) 
X 
X 
X 
X 
X 
X 
X 
Note: AE=adverse event, ET=early termination, M=Month, MAAE=medically attended adverse event, SAE=serious adverse event, V=visit, YF=yellow fever 
Footnotes: 
(a) If the subject terminates early, Day 360 (M12) procedures should be performed. 
(b) Prior to the subject entering into the trial, and before any protocol-directed procedures are performed. 
(c) Review of inclusion/exclusion criteria will be performed prior to the first trial vaccination at Day 1 (M0). After written informed consent has been obtained and 
eligibility is assessed, subjects will be randomized to one of the 3 trial groups. 
(d) Any other vaccination against any flavivirus (licensed or investigational, including any other dengue vaccine) during the entire trial period, all concomitant 
medications and vaccine history from 1 month (minimum 28 days) prior to administration of each dose of trial vaccine(s) up to 1 month (minimum 28 days) 
thereafter, steroids and immunostimulants within 60 days prior to Day 1 (M0), immunoglobulins and blood products within 3 months prior to Day 1 (M0), and 
immunosuppressive therapy within 6 months prior to Day 1 (M0). 
(e) Physical examination at Day 1 (M0) including measurement of weight and height; BMI will be calculated automatically. Measurement of height is only required at 
Day 1 (M0). 
(f) Subjects may undergo a brief symptom-directed physical examination. Clinically significant changes from the Baseline examination should be recorded in the 
subject’s source documents and electronic Case Report Form (CRF). 
(g) Vital signs including (but not limited to) the measurement of systolic blood pressure/diastolic blood pressure, heart rate, and body temperature. 
(h) Pregnancy testing (serum or urine) for females of childbearing potential. Results must be confirmed and documented as negative prior to each vaccination. 
Additional pregnancy tests may be performed during the trial if deemed necessary by the Investigator. 
(i) Females of childbearing potential who are sexually active will be provided with information on acceptable methods of contraception and will be asked prior to 
vaccination on Day 1 (M0) to sign the informed consent form stating that they understand the requirements for avoidance of pregnancy and donation of ova. 
Subjects will be reminded during trial visits to adhere to acceptable contraceptive methods and not donate ova up to 6 weeks post last trial vaccination at Day 180 
(M6). 
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Statistical Analysis Plan (Final, Version 1.0) 
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CONFIDENTIAL 
(j) The blood sample on Day 1 (M0), Day 90 (M3) and Day 180 (M6) should be taken prior to trial vaccination. The blood sample on Day 120 (M4) and Day 210 
(M7) should be taken at least 29 days after the vaccination on Day 90 (M3) and Day 180 (M6), respectively. 
(k) After each trial vaccination, the subject will be observed for at least 30 minutes including observation for solicited local (injection site) and systemic AEs, 
unsolicited AEs (non-serious and serious), and body temperature measurement. 
(l) Diary (paper or electronic) cards will be used for the collection of:  
1) Solicited local (injection site) AEs for 7 days after each vaccination (including the day of vaccination). If solicited local AEs continue on Day 8 following each 
trial vaccination, record the extended information on the Adverse Event CRF. 
2) Solicited systemic AEs for 14 days after each vaccination (including the day of vaccination). If solicited systemic AEs continue on Day 15 following each trial 
vaccination, record the extended information on the Adverse Event CRF. 
The Investigator will categorize events by severity (mild, moderate or severe) and will assess causality to vaccine administration for solicited systemic events 
(related or not related). 
(m) Unsolicited AEs for 28 days (including the day of vaccination) after each vaccination will be collected by interview and recorded for all subjects at Day 30 (M1), 
Day 120 (M4) and Day 210 (M7). The Investigator will categorize events by severity (mild, moderate or severe) and will assess causality to vaccine administration 
(related or not related). 
(n) AEs leading to discontinuation or withdrawal, MAAEs and SAEs will be collected for the trial duration. The Investigator will categorize events by severity (mild, 
moderate or severe) and will assess causality to vaccine administration (related or not related). 
 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
Appendix B Solicited Local (Injection Site) and Systemic Adverse Events and Severity 
Table 8.b 
Solicited Local (Injection Site) and Systemic AEs 
Solicited local (injection site) AEs: 
Pain 
Erythema 
Swelling 
Solicited systemic AEs: 
Fever (a) 
Headache 
Asthenia 
Malaise 
Myalgia 
(a)  Fever is defined as a body temperature ≥38°C (100.4°F) regardless of the method used [7]. 
 
 
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Trial No. DEN-305 
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Statistical Analysis Plan (Final, Version 1.0) 
21 October 2019
 
 
Table 8.c 
Severity of Solicited Safety Parameters 
Adverse Event 
Severity grade 
Severity 
Pain at injection site 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity with or without treatment 
3 
Severe: Prevents daily activity with or without treatment 
Erythema at injection 
site (a) 
0 
<25 mm 
1 
Mild: >25 – ≤50 mm 
2 
Moderate: >50–≤100 mm 
3 
Severe: >100 mm 
Swelling at injection 
site (a) 
0 
<25 mm 
1 
Mild: >25 – ≤50 mm  
2 
Moderate: >50-≤100 mm 
3 
Severe: >100 mm 
Headache 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity with or without treatment 
3 
Severe: Prevents normal activity with or without treatment 
Asthenia 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Malaise  
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Myalgia 
0 
None 
1 
Mild: No interference with daily activity 
2 
Moderate: Interference with daily activity 
3 
Severe: Prevents daily activity 
Fever (b) 
 
NA 
None 
NA 
38.0-<38.5°C 
NA 
38.5-<39.0°C 
NA 
39.0-<39.5°C 
NA 
39.5-<40.0°C 
NA 
40.0-<40.5°C 
NA 
40.5-<41.0°C 
NA 
≥41.0°C 
NA = not applicable 
(a) Subjects are to record greatest surface diameter in mm on the diary card. 
(b) Fever is defined as a body temperature ≥38°C (100.4°F) regardless of the method used [7]. 
 
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