A Phase 1 Clinical Trial to Evaluate the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults
作者: 来源: 日期: 2015-03-14
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Protocol Cover Page

Brief Title:

A Phase 1 Clinical Trial to Evaluate the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults

Protocol Title:

A Phase 1 Double-blind, Dose-escalation, Placebo-controlled Clinical Trial to Evaluate the Safety, Tolerability and Immunogenicity of the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults aged between 18 and 60 years in China.

Protocol Number:

JSVCT020

ClinicalTrials.gov

NCT02326194

Protocol Date:

December 29, 2014

Version:

Version 1.2(final)

Phase:

Phase 1

Sponsor:

Beijing Institute of Biotechnology

 

Address: 20 East Street, Fengtai District, Beijing

 

Tianjin CanSino Biotechnology Inc.

Address: 185 South Ave., TEDA West District, Tianjin

Principle Investigator

Feng-Cai Zhu MSc

Leading Authors

Feng-Cai Zhu

Jiangsu Provincial Center For Disease Control and Prevention

 

Wei Chen

Beijing Institute of Biotechnology

Other Authors

Li-Hua Hou

Beijing Institute of Biotechnology

 

Jing-Xin Li

Jiangsu Provincial Center for Disease Control and Prevention

 

Yue-Mei Hu

Jiangsu Provincial Center for Disease Control and Prevention

 

Fan-Yue Meng

Jiangsu Provincial Center for Disease Control and Prevention

 

Gui-Rong Zhang

Beijing Institute for Drug and Instrument Quality Control

 

Pei Liu

Southeast University

 

Rong Tang

Jiangsu Provincial Center for Disease Control and Prevention

This document contains confidential information belonging to Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc.


DOCUMENT HISTORY

Version No.

Version Date

Amendment

1.0

December 15, 2014

N/A

1.1

December 23, 2014

1st Amendment

1.2

December 29, 2014

2nd Amendment

Information of the 1st Amendment

Contents in Original Version (1.0)

Contents in Altered Version (1.1)

Page/Row

Original Contents

Page/Row

Altered Contents

Row 3 of Page 32 and Row 9 of Page 35

experimental vaccine

Row 3 of Page 32 and Row 9 of Page 35

investigational vaccine

Information of the 2nd Amendment

Contents in Original Version (1.1)

Contents in Altered Version (1.2)

Page/Row

Original Contents

Page/Row

Altered Contents

Row 3 of Page 23

A body mass index (BMI) 18.5-30

Row 4 of Page 23

A body mass index (BMI) <35

Row 9 of Page 31

Screening will take place within 3 days before vaccination.

Row 4 of Page 31

Screening will take place within 5 days before vaccination.

Row 4 of Page 33

First 60 participants in the low dose group will be followed for safety up to 7 days. All the adverse reaction during the first 7 days will be referred to the Data and Safety Monitoring Board (DSMB) to assess whether or not the safety data could support the study moving on to the next stage to test the high dose vaccine.

Row 23 of Page 32

For the low dose group, no more than 5 participants per day will be enrolled for the first 2 days, and no more than 10 participants per day will be enrolled for the next 2 days. At least five of them have been followed for a minimum of 7 days before proceeding to immunize 60 participants in the high dose group.

Row 18 of Page 49

Statistics only submit interim statistical report to the investigator and the sponsor, the unblinded data will not be submitted and clinical trial sites still remain in blinded.

Row 13 of Page 49

The un-blinding data will not be confidential and therefore the clinical trial site will not be kept in blind.

           

CLINICAL TRIAL CONTACT LIST

Sponsor

 

Beijing Institute of Biotechnology

No. 20 East Street, Fengtai District, Beijing 100039, People's Republic of China

Tianjin CanSino Biotechnology Inc.

No. 185 South Ave., TEDA West District, Tianjin 300457, People's Republic of China

Tel:

Fax:

E-mail:

010-63818253

010-63818253

cw0226@foxmail.com

Principal Investigator

Feng-Cai Zhu

Jiangsu Provincial Center for Diseases Control and Prevention

No. 172, Jiangsu Road, Nanjing 210009, Jiangsu Province,

People's Republic of China

Tel:

Fax:

E-mail:

+86-25-83759418

+86-25-83759409

jszfc@vip.sina.com

Contract Research Organization

Yuan Yong

Shenzhen Yingheyuan Medical Technology Development Co., Ltd.

6B-01, Tian-xia Industrial Park, Ma Jia Long, Yi-yuan Road, Nan-shan District , Shenzhen 518052,

People's Republic of China

Tel:

Fax:

E-mail:

+86-755-61373860

+86-755-61373008 ext 602

13382030658@189.cn

Serious Adverse Event

Contact (Sponsor)

Wei Chen

Beijing Institute of Biotechnology

No. 20 East Street, Fengtai District, Beijing 100039, People's Republic of China

Tel:

Fax:

Mob. Phone:

E-mail:

+86-10-63818253

+86-10-63818253

+86-13910789661

cw0226@foxmail.com

Clinical Laboratory

Gui-Rong Zhang

Beijing Institute for Drug and Instrument Quality Control

People's Republic of China

Tel:

Fax:

E-mail:

15901233200

010-63896955

guirong_zhang@126.com

Statistical Party

Pei Liu

Public Health Department, Southeast University,

No. 78 Dingjia Bridge, Nanjing 210009, Jiangsu Province,

People's Republic of China

Tel:

Fax:

E-mail:

+86-25-83272584

+86-25-83272584

liupeiseu@126.com

         

 

PROTOCOL SUMMARY

Brief Title

A Phase I Clinical Trial to Evaluate the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults

Official Title

A Phase 1 Double-blind, Dose-escalation, Placebo-controlled Clinical Trial to Evaluate the Safety, Tolerability and Immunogenicity of the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults aged between 18-60 years in China.

Objectives

Preliminarily evaluate the safety, tolerability and immunogenicity of the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV)

Target population

Healthy adults aged between 18 and 60 years

Sample size

120 participants

Rational and background

Ebola viruses (EBOVs) are enveloped, non-segmented, negative-strand RNA viruses belonging to the family Filoviridae. They are known to cause lethal hemorrhagic fever in humans and non-human primates with a mortality rate up to 90%. EBOVs transmit among humans through closely contact with the infected blood, bodily fluids or tissues; moreover, the intentional release of EBOVs would probably result in mucosal infection by small-particle aerosol dispersion.

Five different species of EBOV have been identified: Zaire; Sudan; Ivory Coast; Reston;

and the newly discovered Bundibugyo. Among these species, infections with Zaire is the most serious and has caused the greatest number of deaths.

The EBOV envelope glycoprotein (EBOV-GP) forms spikes on the surface of mature virions, and has been shown to be an effective target for vaccine design. Preclinical studies indicated that both humoral and cellular responses to EBOV are very important for viral control and clearance.

The Ad5-EBOV is a recombinant replication defective human recombinant Ad5 vector based vaccine expressing Ebola virus glycoprotein (GP) (Guinea, 2014). It is manufactured through cell amplification, purification, formulation with stabilizer and lyophilization. The final product is lyophilized whitepowder.

This is a phase 1 clinical trial. We are going to evaluate the safety, tolerability and immunogenicity (including humoral and cellular response) of the Ad5-EBOV in healthy adults in China.

Investigational vaccines

Experimental Vaccine (Ad5-EBOV): lyophilized white powder

Low dose: 4×1010 virus particle/1 ml

High dose: 1.6×1011 virus particle/2 ml

Placebo: lyophilized-dried powder

Same formulation of excipients as vaccine but without any virus particle.

Immunization schedule

Low dose group:

One shot, 1 ml per dose, intramuscular injection at day 0.

High dose group:

Double shots, 1 ml per dose, one shot in each arm, intramuscular injection at day 0.

Trial design

This is a single center, double-blind, placebo control, dose-escalation clinical trial.

According to the Chinese guidelines for vaccine clinical trial, the sample size of a phase 1 clinical trial should be at least 20. In this study, a total of 120 participants will be included. 60 participants will be firstly recruited and randomly assigned to receive the low dose Ad5-EBOV or placebo in a ratio of 2:1. After the safety of the low dose vaccination is confirmed, another 60 participants will be recruited and randomly assigned to receive the high dose Ad5-EBOV or placebo in a ratio of 2:1. Thus, 40 participants will receive the low dose vaccine, 40 will receive the high dose vaccine and 40 will receive the placebo.

Randomization list will be generated by independent statistician in the School of Public Health, Southeast University, using SAS 9.3 software. Blinding will be maintained for all participants and investigators and their study staff participating in this study.

The whole follow-up period for each participant will be 6 months.

Primary Endpoints

Safety:

-     Occurrence of solicited adverse reactions within 7 days after vaccination.

Immunogenicity:

-     ELISA antigen-specific assays for antibody responses on day 0 and 28.

-     Intracellular cytokine staining (ICS) assay for T cell responses on day 0 and 28.

Secondary Endpoints

Safety:

-     Occurrence of unsolicited adverse reactions within 28 days after vaccination.

-     Change from baseline for safety laboratory measures (hemoglobin, white blood cell count, total lymphocyte count, platelets, creatinine, alanine transaminase, prothrombin time, and activated partial thromboplastin time) on day 3, 14, and 28.

-     Occurrence of serious adverse reaction during the whole follow-up period (6 months).

Immunogenicity:

-     ELISA antigen-specific assays for antibody responses on day 168.

-     Neutralizing antibody titers response to human Ad5 on day 0, 28 and 168.

-     Intracellular cytokine staining (ICS) assay for T cell responses on day 168.

Exploratory Endpoints

-     Change from baseline for safety laboratory measures (hemoglobin, white blood cell count, total lymphocyte count, platelets, creatinine, alanine transaminase, prothrombin time, and activated partial thromboplastin time) on day 56.

-     ELISA antigen-specific assays for antibody responses on day 3, 7, 14, 56 and 112.

-     Neutralizing antibody titers response to human Ad5 on day 3, 7, 14, 56 and 112.

-     Intracellular cytokine staining (ICS) assay for T cell responses on day 7, 14, 56 and 112.

-     T-cell responsiveness by an enzyme-linked immunospot (ELISpot) assay on day 0, 28, and 168.

-     vaccine-induced mRNA expression profiles after vaccination on day 3, and 7.

-     HLA type.

Scheduled site visits

Visit 0 (within 5days before vaccination): informed consent, screening, physical examination, blood sample taking for baseline laboratory measures.

Visit 1 (day 0): blood sample taking for baseline antibody response and cellular response, vaccination, and 6 hours safety observation after injection at site.

Visit 2 (day 3): assessment of safety, blood sample taking for post-vaccination antibody response and safety laboratory measures

Visit 3 (day 7, +1 day): assessment of safety, blood sample taking for post-vaccination antibody response and cellular response.

Visit 4 (day 14,±2 days): assessment of safety, blood sample taking for safety laboratory measures, post-vaccination antibody response and cellular response.

Visit 5 (day 28,±3 days): assessment of safety, blood sample taking for safety laboratory measures, post-vaccination antibody response and cellular response.

Visit 6 (day 56,±5 days): assessment of safety, blood sample taking for safety laboratory measures, post-vaccination antibody response and cellular response.

Visit 7 (day 112,±10 days): assessment of safety, blood sample taking for post-vaccination antibody response and cellular response.

Visit 8 (day 168,±14 days): assessment of safety, blood sample taking for post-vaccination antibody response and cellular response.

Inclusion Criteria

-          Aged between 18 and 60 years.

-          Able to understand the content of informed consent and willing to sign the informed consent

-          Able and willing to complete all the secluded study process during the whole study follow-up period (about 6 months).

-          A body mass index (BMI) <35

-          Hemoglobin 110-150g/L for female, and 120-160g/L for male.

-          White blood cells (WBC) 4.0-10.0×109 cells/L

-          Total lymphocyte Count 0.8-4.5×109 cells/L

-          Platelets 100–300×109 cells/L

-          Alanine aminotransferase (ALT) 0-40U/L

-          Serum creatinine 44-106μmol/L

-          Active partial thromboplastin time (aPTT) 20-40 seconds

-          Prothrombin time (PT) 10-14 seconds

-          Negative in HIV diagnostic blood test

-          Axillary temperature ≤37.0°C on the day of enrollment

-          General good health as established by medical history and physical examination.

Exclusion Criteria

-          Family history of seizure, epilepsy, brain or mental disease

-          Participant that has a medical history of any of the following: allergic history of any vaccination or drugs, or allergic to any ingredient of the Ad5-EBOV, such as mannitol

-          Woman who is pregnant, breast-feeding or positive in β-HCG (human chorionic gonadotropin) pregnancy test (urine) on day of enrollment, or become pregnant during the next 6 months

-          Any acute fever disease or infections in last 7 days

-          Major congenital defects or not well-controlled chronic illness, such as asthma, diabetes, or thyroid disease

-          Hereditary angioneurotic edema or acquired angioneurotic edema

-          Urticaria in last one year

-          Asplenia or functional asplenia

-          Platelet disorder or other bleeding disorder may cause injection contraindication

-          Faint at the sight of blood or needles.

-          Prior administration of immunodepressant or corticosteroids, antianaphylaxis treatment, cytotoxic treatment in last 6 months

-          Prior administration of blood products in last 4 months

-          Prior administration of other research medicines in last 1 month

-          Prior administration of attenuated vaccine in last 1 month

-          Prior administration of inactivated vaccine in last 14 days

-          Current anti-tuberculosis prophylaxis or therapy

-          Any condition that in the opinion of the investigators may interfere with the evaluation of study objectives

Role of the sponsor

Sponsors participate in the trial design and the protocol writing, but will not participate in other process of the trial, including data collection, statistical analysis, data interpretation and writing study report.

Dose escalation

For the low dose group, no more than 5 participants per day will be enrolled for the first 2 days, and no more than 10 participants per day will be enrolled for the next 2 days. At least five of them have been followed for a minimum of 7 days before proceeding to immunize 60 participants in the high dose group. All the adverse reaction will be referred to the Data and Safety Monitoring Board (DSMB) to assess whether or not the safety data can support the study moving on to the next stage. For the high dose group, no more than 5 participants per day will be enrolled for the first 2 days, and no more than 10 participants per day will be enrolled for the next 2 days. The adverse reaction within the first 7 days after the vaccination with high dose vaccine will be updated to the DSMB.

Criteria for pausing or early termination

DSMB will review the reported safety data in the participants for the first 7 days after vaccination. During the study period, if an increase of risk for participants is noticed, the DSMB should promptly inform the principle investigator and sponsors. Sponsors, investigators and DSMB will have a panel meeting, and then DSMB will make final decision to pause or all an early termination of the study.

Administration of study injections and new enrollments will be paused, if:

-          One serious adverse event may be associated with vaccination, or

-          Occurrence of grade 3 adverse reaction may be associated with vaccination in 10% of participants or more (including injection-site reaction, systemic reaction, and change of the safety laboratory measures).

The study may come to an early termination, if:

-          One vaccination-associated serious adverse event, or

-          Occurrence of grade 3 adverse events associated with vaccination in 15% of participants or more (including injection-site reaction, systemic reaction, and change of the safety laboratory measures), or

-          Required by sponsor, or

-          Required by regulatory authority, or

-          Required by institutional review board (IRB).

Interim analyses

Interim analyses will be conducted by the independent statistical party after the last participant completes Visit 5 on day 28, and all the safety data and immunogenicity data up to day 28 will becollected.The data will be unblinded by representatives of statistical party, sponsor, clinical research investigator and CRO. The un-blinding data willnot be confidential andtherefore the clinical trial site will not be kept in blind.

The results will not influence the conduct of the trial in terms of early termination or later safety or immunogenicity endpoint assessments.

Final analyses

Final analyses will be conducted by the independent statistical party. After the last participant completes Visit 8 on day 168, and all the safety data and immunogenicity data up to day 168 are collected, all the data will be reviewed and then locked for the final statistical analysis.

 


 

TABLE OF CONTENTS

 

DOCUMENT HISTORY. 1

CLINICAL TRIAL CONTACT LIST. 2

PROTOCOL SUMMARY. 4

ABBREVIATIONS. 14

1.      BACKGROUND AND RATIONALE. 16

1.1             Introduction. 16

1.2             Ebola virus and disease background. 16

1.3             Vaccine development in other countries. 17

1.4             Rationale of the development of the candidate vaccine Ad5-EBOV in China. 18

2.      PRECLINICAL STUDIES WITH CANDIDATE AD5-EBOV. 19

2.1             Preclinical safety evaluation. 19

2.1.1         Acute toxicity test in mice. 19

2.1.2         Repeated intramuscular injection toxicity test in cynomolgus monkeys. 19

2.2             Immunogenicity of Ad5-EBOV.. 20

2.2.1         Immunogenicity in mice. 20

2.2.2         Immunogenicity in Guinea pigs. 20

2.2.3         Immunogenicity in cynomolgus monkeys. 20

2.3             Summary for preclinical studies. 21

3       STUDY OBJECTIVES AND ENDPOINTS. 21

3.1             Objectives. 21

3.2             Primary endpoints. 21

3.3             Secondary endpoints. 22

3.4             Exploratory endpoints. 22

4       PARTICIPANTS SELECTION.. 22

4.1             Inclusion criteria. 22

4.2             Exclusion criteria. 23

4.3             Withdraw from the study. 24

5       STUDY DESIGN.. 24

5.1             Study design and method description. 24

5.2             Duration of study. 24

5.3             Sample size. 25

5.4             Treatment allocation. 25

5.4.1         Treatment allocation. 25

5.4.2         Blinding. 25

5.4.3         Method of unblinding and breaking the study blind. 26

5.5             Study setting. 26

5.6             Study Procedures. 27

5.6.1         Planned visits in this trial 27

5.6.2         Informed consent 29

5.6.3         Eligibility assessment 31

5.6.4         Randomization and injection administration. 31

5.6.5         Safety follow-up. 32

5.6.6         Criteria for dose escalation. 32

5.6.7         Criteria for pausing the study or an early termination. 33

6       INVESTIGATIONAL PRODUCT. 33

6.1             Experimental vaccineAd5-EBOV.. 33

6.2             Placebo. 35

6.3             Package of the investigational vaccine. 35

6.4             Administration. 36

6.5             Transportation and Storage. 36

7       SAFETY ASSESSMENTS AND ADVERSE EVENT REPORTING.. 37

7.1             Adverse event and adverse reaction. 37

7.2             Grading for adverse events. 38

7.2.1         Injection site. 38

7.2.2         Systemic reactions. 39

7.2.3         Safety laboratory measures. 41

7.3             Serious adverse event/reaction (SAE). 42

7.3.1         Reporting SAEs. 43

7.3.2         Expedited reporting of SUSARs to regulatory authorities and investigators. 43

7.4             Safety data monitoring by DSMB.. 44

7.5             Withdrawal due to adverse events (see also section on participant withdrawal). 45

8       LABORATORY ASSAY AND IMMUNOGENICITY ASSESSMENTS. 45

8.1             Antigen-specific antibody responses. 45

8.2             Antigen-specific CD4+, CD8+ T cell responses. 45

8.3             Neutralizing antibody titers response toAd5. 46

9.      DATA COLLECTION AND MANAGEMENT. 46

9.1             Source documents and source data. 46

9.2             Clinical data management 47

9.3             SAE data management 47

9.4             Archiving. 48

9.5             Database creation and data entry. 48

9.6             The database lock. 49

10.             STATISTICS PLAN AND STATISTICAL ANALYSIS. 49

10.1          Interim analysis. 49

10.2          The final analysis. 49

10.3          Analyzed data sets definition. 49

10.3.1       Data set for safety evaluation. 49

10.3.2       Data set for immunogenicity evaluation. 50

10.3.3       Statistical methods. 50

11.             ETHICAL AND LEGAL ISSUES. 51

11.1           Guideline. 51

11.2           Institutional Review Board. 51

11.3           Ethical Conduct of the Study. 51

11.4           Protocol Amendments and Administrative changes. 52

11.5           Confidentiality of Data and Access to Participant Records. 53

11.6           Benefits and risks. 53

12.             FINANCIAL CONTRACT AND INSURANCE COVERAGE. 54

13.             PUBLICATION OF STUDY RESULTS. 54

14.             APPENDIX. 55

Appendix 1 Informed consent form for the participants in low dose group (Version1.2). 55

Appendix 2 Informed consent form for the participants in high dose group (Version1.2). 62

15.             SIGNATURES. 69

16.             REFERENCE. 70

 

 


 

ABBREVIATIONS

AE

Adverse Event

Ad5

Replication Defective Human Adenovirus Serotype 5

ALT

Alanine Aminotransferase

AR

Adverse Reaction

APTT

Active Partial Thromboplastin Time

BMI

Body Mass Index

CDC

Center For Disease Control And Prevention

CFDA

China Food And Drug Administration

CI

Confidence Interval

CRF

Case Report Form

CRO

Contract Research Organization

CTA

Clinical Trial Authorization

DSMB

°C

Data and Safety Monitoring Board

Degrees Celsius

EBOV

Species Zaire Ebolavirus

ELISA

Enzyme Linked Immunosorbent Assay

ELISpot

Enzyme-Linked Immunospot Assay

HLA

Human Leukocyte Antigen

HIV

Human Immunodeficiency Virus

ICS

Intracellular Cytokine Staining Assay

ITT

Intention To Treat

GCP

Good Clinical Practice

GLP

Good Laboratory Practice

GMT

Geometric Mean Titer

GP

Glycoprotein

ICF

Informed Consent Form

ICH

International Conference On Harmonization (Technical

Requirements For Registration Of Pharmaceuticals For Human Use)

IRB

Institutional Review Board

mRNA

Messenger RNA

NT

Neutralization Antibody

QA

Quality Assurance

PBMC

Peripheral Blood Mononuclear Cells

PT

Prothrombin Time

P.R. China

People’s Republic Of China

SAE

Serious Adverse Event

SOP

Vp

Standard Operation Procedure

Viral Particle

WHO

World Health Organization

 


 

1.         BACKGROUND AND RATIONALE

1.1     Introduction

The candidate Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV) against Ebola disease is developed by Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc. This is a phase 1 clinical trial. We are going to evaluate the safety, tolerability and immunogenicity (including humoral and cellular immune response) of the Ad5-EBOV in healthy adults aged between 18 and 60 years in China.

The candidate Ad5-EBOV is a recombinant replication defective recombinant Ad5 vector based vaccine expressing Ebola virus GP (Guinea, 2014). In preclinical studies, the Ad5-EBOV showed a good safety and immunogenicity profile. The Ad5-EBOV has been approved for clinical trials in December 12, 2014 (approval number: 2014JTL005). This protocol has been made according to Good Clinical Practice (GCP), the Declaration of Helsinki, and local rules and regulations of China.

1.2     Ebola virus and disease background

Ebola viruses (EBOVs) are enveloped, non-segmented, negative-strand RNA viruses belonging to the family Filoviridae.1,2 They are known to cause lethal hemorrhagic fever in humans and non-human primates with a mortality rate up to 90%.3,4 Ebola virus disease (EVD), formerly known as Ebola haemorrhagic fever, is a severe, often fatal illness in humans. EBOVs transmit among humans through closely contact with the infected blood, bodily fluids or tissues; moreover, the intentional release of EBOVs would probably result in mucosal infection by small-particle aerosol dispersion.5

The current outbreak in west Africa, (first cases notified in March 2014), is the largest and most complex Ebola outbreak since the Ebola virus was first discovered in 1976.6 There have been more cases and deaths in this outbreak than all others combined. It has also spread between countries starting in Guinea then spreading across land borders to Sierra Leone and Liberia, by air (1 traveller only) to Nigeria, and by land (1 traveller) to Senegal.

The most severely affected countries, Guinea, Sierra Leone and Liberia have very weak health systems, lacking human and infrastructural resources, having only recently emerged from long periods of conflict and instability. On August 8, the WHO Director-General declared this outbreak a Public Health Emergency of International Concern.7

Up to now, five different species of EBOV have been identified: Zaire; Sudan; Ivory Coast; Reston; and the newly discovered Bundibugyo.8 Among these species, infections with Zaire is the most serious and has caused the greatest number of deaths. The virus causing the 2014 West African outbreak belongs to the Zaire species.3

1.3     Vaccine development in other countries

There is as yet no proven treatment available for EVD. However, a range of potential treatments including blood products, immune therapies and drug therapies are currently being evaluated. No licensed vaccines are available yet, but several potential vaccines are undergoing human safety testing.

The EBOV envelope glycoprotein (EBOV-GP) forms spikes on the surface of mature virion, and has been shown to be an effective target for vaccine design.

Table 1 Summary of the registered studies for evaluation of Ebola Vaccines

Study Identifier

(Clinicaltrials.gov)

Status

Design

Vaccine

Age

Sample

Size

NCT00997607

Completed

Phase 1b; double blind

Ebola DNA Plasmid Vaccine

18 to 50 years

30

NCT00605514

Completed

Phase 1; open

Ebola DNA Plasmid Vaccine

18 to 60 years

20

NCT00072605

Completed

Phase 1; randomized

Ebola DNA Plasmid Vaccine

18 to 44 years

27

NCT02240875

Ongoing, but not recruiting

Phase 1; open

Vaccine cAd3-EBOZ

18 to 50 years

60

NCT00374309

Completed

Phase 1; controlled, blinded

Ebola adenovirus serotype 5 vector

18 to 50 years

48

NCT02280408

Recruiting

Phase 1

Prime-Boost VSV Ebola Vaccine

18 to 65 years

120

NCT02296983

Not yet open

Phase 1; open

VSV-ZEBOV vaccine

18 to 55 years

40

According the preliminary report of the clinical trial (NCT02240875) with a Chimpanzee Adenovirus Vector Ebola

Vaccine, no safety concerns were identified. But transient fever developed within 1 day after vaccination in two participants who had received the 2×1011 particle-unit dose. However, data in this trial indicated that the reactogenicity and immune responses to cAd3-EBO vaccine were dose-dependent, and the 2×1011 particle-unit dose was considered to be more immunogenic. In addition, in the clinical research of VSV vector vaccine (NCT02280408), investigators have noticed that vaccination may cause joint pain in the participants. But the symptoms will release several days later.

Replication defective adenovirus vectorsare rapidly cleared from the host. They induce innate immune system responses and through effective protein expression and intracellular production of the vaccine antigen,induce adaptive antibody, CD4+ T cell, and CD8+ T cell responses. There is theoretical concern that pre-existing immunity to viral vectorsmay impact vaccine efficacy which had been shown in the previous clinical trial with a Ad5 Ebola vaccine (NCT00374309). Humoral responses seemed to be negatively impacted by pre-existing Ad5 seropositivity. But the vaccine was well tolerated by participants in both dose groups regardless of pre-existing  Ad5 antibody status.9

Besides, previous adenovirus vaccines in other countries had reported that adenovirus as vectors may cause an increase in bleeding time in a short period of time after vaccinations.10But it won’t last and generally is not life-threatening.Thus, several adenoviral vector vaccines have been approved for market in other countries.

Because there is no other Ebola vaccines are available in China, so no positive comparator could be found for this trial. We will use placebo as the control comparator.

1.4     Rationale of the development of the candidate vaccine Ad5-EBOV in China

Since its first outbreak occurred in 1976, Zaire Ebola virushave been associated with 14 outbreaks reported up to 2014. The Zaire Ebola virus in 2014 causing the most serious outbreak was considered to be a new epidemic strain, with GP homology of the gene was only 97.6%, compared to the GP gene of the strain in 1976.11All the other Ebola Vaccines undergoing the clinical trial were developed based on the Zaire-Mayinga (1976 strain). This experimental Ad5-EBOV is the first Ebola vaccine developed according to the 2014 epidemic strain. Besides, the Ad5-EBOV are lyophilized white products which could be stored at 2-8°C. The lyophilized formulation of the vaccine may be more suitable for use in some area where the cold chain system is incomplete.

2.         PRECLINICAL STUDIES WITH CANDIDATE AD5-EBOV

2.1     Preclinical safety evaluation

2.1.1          Acute toxicity test in mice

The acute effects of the candidate Ad5-EBOV had been examined in Kunming mice. Mice were assigned to a control group or a treatment group, with ten (five female and five male) in each group. Mice in the treatment group were injected intramuscularly a maximum single-dose of Ebola vaccine at 4×1012vp/kg, and the control mice received the same volume of excipient solution without virus particles. The acute toxic reactions were observed and recorded for 14 days after dosing. General clinical observations: no animal death was observed in any group after dosing in the 2 weeks, and there were no abnormalities in animal general clinical signs, including behavior, stool appearance and color, skin, mucous membranes, breathing, heartbeat, fur, eyes, nose and limbs. Animals in the treatment group showed stable growth in body weight, and there was no significant difference as compared to the control group. Mice were sacrificed for gross necropsy at the end of the observation period, and no obvious abnormalities in the main organs and tissues were observed in both groups. In summary, in this experimental condition, the maximum tolerated doseof single intramuscular injection of recombinant Ebola virus vaccine is greater than 4×1012vp/kg for the mice (equivalent to 1500 times of human highest clinical dose in body weight).

2.1.2          Repeated intramuscular injection toxicity test in cynomolgus monkeys

The safe dosage of the Ad5-EBOV was examined in the cynomolgus monkey. Monkeys were assigned to three groups: control group, 2×1010vp group, 2×1011vp group, 6 cynomolgus monkeys (three femaleand three male) of each group. Two dosing of the vaccine or excipient was planned to be given to the animals (intramuscular injection each 4 weeks). After dosing, general clinical observations, body weight, food consumption, body temperature, electrocardiogram, haematology, clinical biochemistry, urine chemistry, immunology parameters and histopathology would be studied. The current results were showed as follows. Animals in each group showed no obvious abnormalities in general clinical observations, body weight, food consumption, body temperature, electrocardiogram, haematology, clinical biochemistry and urine chemistry. Two weeks after dosing, specific serum antibody of the vaccine in low and high groups were both significantly elevated. In conclusion, the current results showed that in this experimental condition, intramuscular injection of the low and high doses of Ad5-EBOV in cynomolgus monkeys did not pose a major risk of toxic effects, and that the monkeys were well tolerated to the vaccine.

2.2     Immunogenicity of Ad5-EBOV

2.2.1          Immunogenicity in mice

Dose:

BalB/C mice were inoculated with 106 ifu or 107 ifu of the Ad5-EBOV and their serum were collected on week 2, 3, 4, 6, 8 and 10 after immunization for IgG ELISA assay. A dose-dependent IgG level was observed and the IgG titer was higher from mice inoculated with high dose.

Inoculation times

BalB/C mice were inoculated with 107ifu of the Ad5-EBOV one time or two times after a 4- week interval. The serum were collected on week 2, 4, 8,12, 16, 24 after immunization for IgG ELISA assay. Serum form mice with two inoculation showed higher IgG titer than those from mice with one inoculation. The IgG titer did not decreased significantly on week 24, which meant one immunization could be used.

T cell response:

BALB/c mice were used for evaluating the immunogenicity of the Ad5-EBOV. ELISpot assay and ICS showed that  Ad5-EBOV can induce rapid and robust cellular immune response.12 At the same time, the IFN-γ positive CD8 cells were gated for detecting the expression of CD107 and the secretion of TNF and IL-2. At the same time, the IFN-γ positive CD8 cells were gated for detecting the expression of CD107 and the secretion of TNF and IL-2. The result showed that about 40% of IFN-γ positive CD8 cells were IFN-γ single positive cells; however, more than 5% IFN-γ, TNF, IL-2 and CD107 tetra-positive cells were detected and the percentage of IFN-γ, TNF and IL-2 tri-positive cells was greater than 10%. This result suggested that a large proportion of the vaccine-induced effector T cells are multifunctional T cells.

2.2.2          Immunogenicity in Guinea pigs

36 guinea pigs were grouped for evaluating the immunogenicity of the Ad5-EBOV, including blank group, 107ifu group and 106ifu group. The IgG titer was assayed 4 weeks after inoculation and 107ifu group showed significantly higher IgG tier, three fold titer of 106ifu group.

2.2.3          Immunogenicity in cynomolgus monkeys

18 cynomolgus monkeys were grouped for evaluating the immunogenicity of the Ad5-EBOV on 2 week and 4 week after immunization, including blank group, 2×1010vp group and 2×1011vp group. The IgG titers reached high level on 2 week and there was no significant difference between 2×1010vp group and 2×1011vp group. Further there was no significant difference for IgG titer between on 2 week and 4 week.

2.3     Summary for preclinical studies

The Ad5-EBOV, developed by Beijing Institute of Biotechnology/Tianjin CanSino Biotechnology Inc., is a replication-defective adenovirus type 5 vaccine which expresses Ebola virus Zaire (Guinea, 2014) envelope glycoprotein. Ebola GP could stimulate humoral and cellular responses to resist Ebola virus infection. The Ad5-EBOV was a replication-defective virus, which could not replicate in vivo. Previous study showed humoral and cellular responses played important role in protection from infection.13 Immunological test of the Ad5-EBOV on animals showed good immunogenicity and safety, which meant the Ad5-EBOV was ready for clinical evaluation.

Based on the Drug Administration Law of China, Jiangsu Provincial Center for Disease Control and Prevention was entrusted to conduct a phase 1 clinical trial for the candidate Ad5-EBOV.

3             STUDY OBJECTIVES AND ENDPOINTS

3.1     Objectives

Preliminarily evaluate the safety, tolerability and immunogenicity (humoral and cellular responses) of the Recombinant Human Type 5 Adenovirus Vector Based Ebola Vaccine (Ad5-EBOV).

3.2     Primary endpoints

Safety:

-          Occurrence of solicited adverse reactions within 7 days after vaccination.

Immunogenicity:

-          ELISA antigen-specific assays for antibody responses on day 0 and 28.

-          Intracellular cytokine staining (ICS) assay for T cell responses on day 0 and 28.

3.3     Secondary endpoints

Safety:

-     Occurrence of unsolicited adverse reactions within 28 days after vaccination.

-     Change from baseline for safety laboratory measures (hemoglobin, white blood cell count, total lymphocyte count, platelets, creatinine, alanine transaminase, prothrombin time, and activated partial thromboplastin time) on day 3, 14, and 28.

-     Occurrence of serious adverse reaction during the whole follow-up period (6 months).

Immunogenicity:

-     ELISA antigen-specific assays for antibody responses on day 168.

-     Neutralizing antibody titers response to human Ad5 on day 0, 28 and 168.

-     Intracellular cytokine staining (ICS) assay for T cell responses on day 168.

3.4     Exploratory endpoints

-     Change from baseline for safety laboratory measures (hemoglobin, white blood cell count, total lymphocyte count, platelets, creatinine, alanine transaminase, prothrombin time, and activated partial thromboplastin time) on day 56.

-     ELISA antigen-specific assays for antibody responses on day 3, 7, 14, 56 and 112.

-     Neutralizing antibody titers response to human Ad5 on day 3, 7, 14, 56 and 112.

-     Intracellular cytokine staining (ICS) assay for T cell responses on day 7, 14, 56 and 112.

-     T-cell responsiveness by an enzyme-linked immunospot (ELISpot) assay on day 0, 14, 28, and 168.

-     vaccine-induced mRNA expression profiles after vaccination on day 3, and 7.

-     HLA type.

4             PARTICIPANTS SELECTION

The following inclusion and exclusion criteria will be used to select the eligible participants for this study.

4.1     Inclusion criteria

-     Aged between 18 and 60 years.

-     Able to understand the content of informed consent and willing to sign the informed consent

-     Able and willing to complete all the secluded study process during the whole study follow-up period (about 6 months).

-     A body mass index (BMI) <35

-     Hemoglobin 110-150g/L for female, and 120-160g/L for male.

-     White blood cells (WBC) 4.0-10.0×109 cells/L

-     Total lymphocyte Count 0.8-4.5×109 cells/L

-     Platelets 100–300×109 cells/L

-     Alanine aminotransferase (ALT) 0-40U/L

-     Serum creatinine 44-106μmol/L

-     Active partial thromboplastin time (APTT) 20-40 seconds

-     Prothrombin time (PT) 10-14 seconds

-     Negative in HIV diagnostic blood test

-     Axillary temperature ≤37.0°C on the day of enrollment

-     General good health as established by medical history and physical examination.

4.2     Exclusion criteria

-     Family history of seizure, epilepsy, brain or mental disease

-     Participant that has a medical history of any of the following: allergic history of any vaccination or drugs, or allergic to any ingredient of the Ad5-EBOV vaccine, such as mannitol

-     Woman who is pregnant, breast-feeding or positive in β-HCG (human chorionic gonadotropin) pregnancy test (urine) on day of enrollment, or become pregnant during the next 6 months

-     Any acute fever disease or infections in last 7 days

-     Major congenital defects or not well-controlled chronic illness, such as asthma, diabetes, or thyroid disease

-     Hereditary angioneurotic edema or acquired angioneurotic edema

-     Urticaria in last one year

-     Asplenia or functional asplenia

-     Platelet disorder or other bleeding disorder may cause injection contraindication

-     Faint at the sight of blood or needles.

-     Prior administration of immunodepressant or corticosteroids, antianaphylaxis treatment, cytotoxic treatment in last 6 months

-     Prior administration of blood products in last 4 months

-     Prior administration of other research medicines in last 1 month

-     Prior administration of attenuated vaccine in last 1 month

-     Prior administration of inactivated vaccine in last 14 days

-     Current anti-tuberculosis prophylaxis or therapy

-     Any condition that in the opinion of the investigators may interfere with the evaluation of study objectives.

4.3     Withdraw from the study

-     Severe violation of the protocol.

-     Have some food and/or medicine can interfere with the immune response during the observation period of the primary objectives (day 0 to day 28).

-     Unwilling to continue the study and request to withdraw.

-     Any intolerable adverse events (related to vaccination or not).

-     New diagnosed disorder which makes the participants not suitable to continue the study.

-     The presence of any condition needs to be withdrawn determined by investigators.

5             STUDY DESIGN

5.1     Study design and method description

This is a single-center, double-blind, dose-escalation, placebo-controlled clinical trial to evaluate the safety, tolerability and immunogenicity of the Recombinant Human Type 5 AdenovirusVector Based Ebola Vaccine (Ad5-EBOV) in healthy adults in China.

5.2     Duration of study

The whole follow-up period for each participant will be 6 months.

5.3     Sample size

According to the Chinese guidelines for vaccine clinical trial, the sample size of a phase 1 clinical trial should be at least 20. We did not calculated the sample size specifically according to statistical power. In this trial, a total of 120 participants will be enrolled. Among them, 40 will receive the low dose vaccine, 40 will receive the high dose vaccine, and 40 will receive the placebo.

Table 2 planned sample size in each group.

Dose groups

Trial vaccine group

Placebo-controlled group

Total

The low dose group

40

20

60

The high dose group

40

20

60

5.4     Treatment allocation

5.4.1          Treatment allocation

Low dose group:

Experimental: injection with 4×1010vp/1ml Ad5-EBOV; one vial of vaccine dissolved(4.0×1010vp/vial) in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

Control: injection with 0vp/1ml placebo; dissolved one vial placebo in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

High dose group:

Experimental: 1.6×1011vp/2ml Ad5-EBOV; two vials of vaccine dissolved (4.0×1010vp/vial) in 1ml sterile water for injection. Doubleshots of 0.8×1011vp/dose will be allocated intramuscularly in two arms at day 0 (one shot in each arm).Thus, participants in the high dose group will be vaccinated with a dose of 1.6×1011vp (=4.0×1010vp/vial× 4).

Control: double shots of 0vp/1 ml placebo; two vials of placebo dissolved in 1ml sterile water for injection. Two injections will be allocated intramuscularly in two arms at day 0 (one shot in each arm).

5.4.2          Blinding

This study will be performed in a double-blinded manner. Blinding will be maintained for all participants and investigators and the study staffs participating in this study. Sponsor’s personnel who directly involved in the conduct of this study (e.g. site monitors, medical monitors, laboratory personnel, Data and Safety Monitoring Board(DSMB), etc.) will also be blinded to the participant’s treatment assignments.

The investigational vaccines and placebo will be randomized and coded by independent statistician from the School of Public Health, Southeast University, People's Republic of China, using SAS 9.3 software. All the randomized vaccine and placebo doses will be re-labeled with randomized numbers which are the sole identifier for each dose. Therefore, all the vaccine and placebo doses are blindly coded. The person who participate in blinding shall not participate in any other process of the clinical trial, also shall not disclose the contents of blinding to any personnel to participate in clinical trial work. The independent statistician should produce a copy of emergency blinding code which should note the title of program and the character “To Be Opened Only in Case of Emergency”.

5.4.3          Method of unblinding and breaking the study blind

During the study, the breaking of the treatment code is forbidden, except in the event of a medical emergency where the investigator believes it is necessary to determine the treatment code in order to initiate appropriate treatment. If knowledge of the treatment code is required, the investigator will open only the specific participant’s code envelope. A signature, date, time and reason will be written on the opened code envelope and the participant with this code has to be withdrawn from this study.

The investigator will assess the relationship between the adverse event and the investigational products before the treatment code is unblinded. The investigator will immediately notify the sponsors at the 24 hours emergency call number (Wei Chen +86-13910789661) when the treatment code is broken on any participant for any reason during the study. The reason for the treatment code being broken must be also documented in the participant’s medical records and in the CRF.

At the end of the study, all code envelopes (intact and opened) must be accounted for and are to be collected by the monitor to be destroyed.

5.5     Study setting

This trial will be conducted in Phase 1 vaccine clinical trial center of Jiangsu Provincial Center for Disease Control and Prevention. Address: Vaccine engineering center, Taizhou Medical New & Hi-tech Industrial Development Zone, Taizhou, Jiangsu Province, China.

5.6     Study Procedures

5.6.1          Planned visits in this trial

Participants will attend a total of 9 visits, including the screening visit (V0) before the vaccination and 8 site visits during a follow-up period of 6 months.

Table 3: 9 scheduled visits for participants in 6-month follow-up

VisitV

Scheduled time point

Visit window

Visit0V0

Within 5days before vaccination

Visit1V1

Vaccination at Day 0

Visit2V2

Day 3 after vaccination

Visit3V3

Day 7 after vaccination

+1 days

Visit4V4

Day 14 after vaccination

±2 days

Visit5V5

Day 28 after vaccination

±3 days

Visit6V6

Day 56 after vaccination

±5 days

Visit7V7

Day 112 after vaccination

±10 days

Visit8V8

Day 168 after vaccination

±14 days

 

Table 4 Detailed information at planned visits in this trial

 

V0

V1

V2

V3

V4

V5

V6

V7

V8

Within 5 days before Day 0

Day0

Day3

Day7

Day 14

Day 28

Day 56

Day

112

Day

168

1

Informed consent

 

 

 

 

 

 

 

 

2

Collecting demographic information

 

 

 

 

 

 

 

 

3

Check inclusion and exclusion criteria

 

 

 

 

 

 

 

4

Physical examination

 

 

Height, weight

 

 

 

 

 

 

 

 

 

Respiratory rate, pulse rate, blood pressure

 

 

 

 

 

 

 

 

 

Urine pregnancy test (female)

 

 

 

 

 

 

 

 

 

Routine blood test

 

 

 

 

 

Blood biochemical test

 

 

 

 

 

aPTT, PT

 

 

 

 

 

HIV test

 

 

 

 

 

 

 

5

Pre-vaccination body temperature

 

 

 

 

 

 

 

 

6

Randomization

 

 

 

 

 

 

 

 

7

Serum analyses

 

 

Antigen-specific antibody response by ELISA

 

 

Neutralizing antibody titers response to human Ad5

 

 

Cellular responses

 

 

ICS

 

 

 

ELISpot

 

 

 

 

 

HLA typing

 

 

 

 

 

 

 

 

mRNA

 

 

 

 

 

 

8

Vaccination

 

 

 

 

 

 

 

 

9

Observation for 30 minutes post-vaccination

 

 

 

 

 

 

 

 

10

Distribution of diary cards

 

 

 

 

 

 

 

11

Post-vaccination recording of solicited / unsolicited symptoms

 

 

 

 

12

Report grade 3 adverse events

 

 

 

 

13

Report serious adverse events

14

Return of diary cards

 

 

 

 

 

 

 

 

15

Record on the “Vaccination and visits record”

 

 

Table 5 Blood taking schedule in this trial(ml)

Visit

 

V0

V1

V2

V3

V4

V5

V6

V7

V8

Time point (Week)

 

Before W0

W0

W1

W1

W2

W4

W8

W16

W24

Time point (Day)

 

Within 5 days before Day 0

Day0

Day3

Day7

Day14

Day28

Day56

Day

112

Day

168

Clinical evaluations

Tube

                 

Routine blood test

 

2

 

2

 

2

2

2

 

 

Blood biochemical test

 

5

 

5

 

5

5

5

 

 

HIV

 

3

 

 

 

   

 

 

3

APTT, PT

 

3

 

3

 

3

3

3

 

 

Antibody assays and serum storage

SST

 

12

12

12

12

12

12

12

12

PBMC and plasma for storage

EDTA

 

40

20

30

30

40

30

30

40

mRNA

PAXgene

 

3

3

3

 

 

 

 

 

Daily Volume

 

13

55

45

45

52

62

52

42

52

Cumulative Volume

 

13

68

113

158

210

272

324

366

418

* Day 0=day of enrollment and vaccine injection. Day 0 evaluations prior to the injection are the baseline for assessing immunogenicity.

5.6.2          Informed consent

In obtaining and documenting informed consent, the investigator should comply with the applicable regulatory requirement(s), and should adhere to GCP and to the ethical principles that have their origin in the Declaration of Helsinki. Prior to the beginning of the trial, the investigator should have the IRB’s written approval/favorable opinion of the written informed consent form and any other written information to be provided to the participants.

Before enrollment and before any study-related procedures are performed, voluntary written study-specific informed consent must be obtained from each participant. A copy of the signed and dated ICF must be given to the participants. The source data must reflect that the informed consent is obtained before participation in the study..

Both the written Informed Consent Form and any other written information to be provided to the participants should include explanations of the following:

-     The purpose of the trial.

-     The trial intervention(s) and the probability for random assignment to each intervention.

-     The trial procedures to be followed, including all invasive procedures.

-     The participant’s responsibilities.

-     All contents related to the nature of the study.

-     The reasonably foreseeable risks or inconveniences to the participants.

-     The reasonable expected benefits. When there is no intended clinical benefit to participants, the participants should be made aware of this.

-     Replacement interventions/vaccines available for selection and its reasonably foreseeable benefits and risks.

-     The compensation and/or treatment available to participants in the event of trial-related injury.

-     That the participation in the trial is voluntary and participants may refuse to participate or withdraw from the trial, at any time, without penalty or loss of benefits to which participants are otherwise entitled.

-     That the monitor(s), the auditor(s), the IRB, and the regulatory authority will be granted direct access to the participants’ original medical records for verification of clinical trial procedures and/or data, without violating the confidentiality of participants, to the extent permitted by the applicable laws and regulations and that, by signing a written informed consent, the participants is authorizing such access.

-     The records for identifying participants will be kept confidential and, to the extent permitted by the applicable laws and/or regulations, will not be made publicly available. If the results of the trial are published, participants’ identity will remain confidential.

-     Offer the contact information of the person(s) to contact for further information regarding the trial and the rights of trial participants, and who to contact in the event of trial-related injury.

-     The expected duration of the trial.

-     The number of participants involved in the trial.

The sponsors will prepare a model Informed Consent Form which will embody all the elements described above. While it is strongly recommended that this model document be followed as closely as possible, the informed consent requirements given in this document are not intended to pre-empt any local regulations which require additional information to be disclosed for informed consent to be legally effective. Clinical judgment, local regulations and requirements should guide the final structure and content of the document.

The investigator has the final responsibility for the final presentation of Informed Consent Form, respecting the mandatory requirements of local regulations. The consent form generated by the investigator with the assistance of the sponsor’s representative, must be approved (along with the protocol, and any other necessary documentation) by the IRB and be acceptable to the sponsors.

5.6.3          Eligibility assessment

Screening will take place within 5 days before vaccination. Prior to inclusion into the study, all participants will be assessed for their eligibility to participate in the study.

The following procedures/assessments will be performed:

-     Compliance with inclusion/exclusion criteria.

-     Assessment of medical history.

-     Recording of demographic data.

-     Baseline laboratory measures (hemoglobin, white blood cell count, total lymphocyte count, platelets, creatinine, alanine transaminase, prothrombin time, and activated partial thromboplastin time).

-     A physical examination including vital signs (height, weight, temperature).

5.6.4          Randomization and injection administration

In this study, a total of 120 participants will be included. 60 participants will be firstly recruited and randomly assigned to receive the low dose Ad5-EBOV or placebo in a ratio of 2:1. After the safety of the low dose vaccination is confirmed, another 60 participants will be recruited and randomly assigned to receive the high dose Ad5-EBOV or placeboin a ratio of 2:1. Thus, 40 participants will receive the low dose vaccine, 40 will receive the high dose vaccine and 40 will receive the placebo.

The injection will be given intramuscularly in the deltoid muscle in participants.

Low dose group:

Experimental: injection with 4×1010vp/1ml Ad5-EBOV; one vial of vaccine dissolved (4.0×1010vp/vial) in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

Control: injection with 0vp/1ml placebo; dissolved one vial placebo in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

High dose group:

Experimental: 1.6×1011vp/2ml Ad5-EBOV; two vials of vaccine dissolved (4.0×1010vp/vial) in 1ml sterile water for injection. Doubleshots of 0.8×1011vp/dose will be allocated intramuscularly in two arms at day 0 (one shot in each arm). Thus, participants in the high dose group will be vaccinated with a dose of 1.6×1011vp (=4.0×1010vp/vial× 4).

Control: double shots of 0vp/1 ml placebo; two vials of placebo dissolved in 1ml sterile water for injection. Two injections will be allocated intramuscularly in two arms at day 0 (one shot in each arm).

5.6.5          Safety follow-up

Safety follow-ups are divided into three stages:

Stage 1: The solicited adverse reaction collecting phase (from day 0 to 7)

Stage 2: The unsolicited adverse reaction collecting phase (from day 8 to day 28)

Stage 3: In the whole study period from day 0 to month 6, serious adverse event (SAE) will be reported and recorded.

After vaccination, each participant will be asked to stay at study site for at least 6 hours safety surveillance. Doctors will monitor the vital signs of the participants and teach them to record any adverse reactions or events on the diary card. If there is no significant adverse reactions, participants will be allowed to go home and record axillary temperature and any adverse events (AEs) for seven consecutive days by their own in a diary card provided by the investigators.

During the first 7 days after vaccination, investigators will collect the safety data from participants daily. Participants returned on day 3 (Visit 2), day 7 (Visit 3) for review of the diary card, concomitant medications, and blood taking for tests. Investigators will give a telephone interview for each participants at least once a day in the rest period (day 1, day 2, day 4, day 5, day 6).

From day 8 to 28 after each vaccination, participants continue to record any unsolicited AEs and concomitant medications into a diary card. Participants are required to return to site at day 14 (Visit 4), day 28 (Visit 5) after vaccination for collection of safety data and blood taking for tests.

5.6.6          Criteria for dose escalation

For the low dose group, no more than 5 participants per day will be enrolled for the first 2 days, and no more than 10 participants per day will be enrolled for the next 2 days. At least five of them have been followed for a minimum of 7 days before proceeding to immunize 60 participants in the high dose group.All the adverse reaction will be referred to the Data and Safety Monitoring Board (DSMB) to assess whether or not the safety data can support the study moving on to the next stage. For the high dose group, no more than 5 participants per day will be enrolled for the first 2 days, and no more than 10 participants per day will be enrolled for the next 2 days. The adverse reaction within the first 7 days after the vaccination with high dose vaccine will be updated to the DSMB.

5.6.7          Criteria for pausing the study or an early termination

DSMB will review the reported safety data in the participants for the first 7 days after vaccination. During the study period, if an increase of risk for participants is noticed, the DSMB should promptly inform the principle investigator and sponsors. Sponsors, investigators and DSMB will have a panel meeting, and then DSMB will make final decision to pause or all an early termination of the study.

Administration of study injections and new enrollments will be paused, if:

-          One serious adverse event may be associated with vaccination, or

-          Occurrence of grade 3 adverse reaction may be associated with vaccination in 10% of participants or more (including injection-site reaction, systemic reaction, and change of the safety laboratory measures).

The study may come to the early termination, if:

-          One vaccination-associated serious adverse event, or

-          Occurrence of grade 3 adverse events associated with vaccination in 15% of participants or more (including injection-site reaction, systemic reaction, and change of the safety laboratory measures), or

-          Required by sponsor, or

-          Required by regulatory authority, or

-          Required by institutional review board (IRB).

6             INVESTIGATIONAL PRODUCT

6.1   Experimental vaccineAd5-EBOV

Experimental vaccine Ad5-EBOV, developed by Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc., is a replication defective Adenovirus Type 5 Vector based vaccine which expresses Ebola virus Zaire (Guinea, 2014) envelope glycoprotein. The final product is lyophilized white powder, with 4.0×1010vp/vial. The antigen contents in the Ad5-EBOV were measured according Ebola vaccine antigen standard by National Institute for Food and Drug Control (NIFDC).

Stability of the vaccine:

The recombinant Ebola vaccine lyophilized formulation should be stored and transported at 2~8°C, away from light. Preliminary stability results in 2 weeks at 37°C showed that virus titer number of virus particles, virus purity, moisture content, pH, bacterial endotoxin test results are all in line with product quality standards, no significant difference was found compared with the initial results at zero time point. This result was also found at 4°C storage conditions for one month. According to the 2010 edition of "Chinese Pharmacopoeia" (Volume III) , the virus titer loss should be less than 1.0 log when stored at 37°C for one week, and the product shelf lifeisgood for18 months. Because there was no significant virus titer loss of this Ebola vaccine under 2 week at 37°C, it is estimated that it can be stored at 2~8°C for at least 12 months.

Vaccine quality research and verification:

We developed quality standards of the EBOV bulk drug substance and the finished product according to the 2010 edition of "Chinese Pharmacopoeia"(Volume III). The clinical trial batch samples 201411001 were tested by the manufacturer Tianjin CanSino Biotechnology and were also confirmed by the NIFDC. All test results meet the quality standards. Table 6 below shows that vaccines self-test results are consistent with the test results of NIFDC.

Table 6 Vaccines quality standard inspection

No.

Test

Specification

201411001

(CanSino)

201411001

(NIFDC)

1

Physical Appearance

White or milky white loose body, after adding the diluents with labeled amount, dissolved as a colorless transparent liquid after shaking , no visible foreign particles.

Conforms with the regulations

Conforms with the regulations

2

Titration on 293 cells(IFU/dose)

≥1×108  IFU/dose

7.2×108

1×109

3

The identification

of  Viral vectors

Amplified fragment size should be consistent with the theory (735bp) by PCR

Conforms with the regulations

Conforms with the regulations

4

pH

7.08.0

7.4

7.2

5

The identification

of target gene

Amplified fragment size should be consistent with the theory (2000bp) by PCR

Conforms with the regulations

Conforms with the regulations

6

Moisture(%)

≤3.0%

2.5

2.4

7

Detection of EBOV  Expression from Infected 293 cells        

The antigen expression should be positive by Western blot method

Conforms with the regulations

Conforms with the regulations

8

Sterility test

Sterility

Conforms with the regulations

Conforms with the regulations

9

Virus particle number (vp/dose)

≥2.0×1010 VP/dose

4.5×1010

4.9×1010

10

Bacterial Endotoxin Detection(EU/dose)

≤50 EU/dose

1.1

3

11

Purity(%)

≥95.0%

98.2%

99.8%

12

Abnormal Toxicity Detection

Animals should be fully healthy within the observation period, and no abnormal reaction, Each animal weight gain maturity.

Conforms with the regulations

Conforms with the regulations

13

Osmolarity(mosm/kg)

400650 mosm/kg

527

529

6.2   Placebo

The placebo is lyophilized-dried powder, contains same formulation of excipients as vaccine, with no virus particle which could express Ebola virus Zaire envelope glycoprotein. The placebo were also measured to be qualified by NIFDC.

In order to maintain blinding, participants in low dose group will be vaccinated with one vial of placebo dissolved in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

For participants in high dose group, each shot contains two vials of placebo dissolved in 1ml sterile water for injection. Double shots will be allocated intramuscularly in two arms at day 0 (one shot in each arm) in order to keep blinding.

6.3   Package of the investigational vaccine

Internal and external packaging of the experimental vaccines is the same as placebos, only with a sequential number as the only identifier. The vaccines will be packed in labeled boxes. Each label will contain the following information: name of vaccine, manufacturing enterprises, vaccine code, expiry date, storage conditions and “FOR CLINICAL TRIAL USE ONLY”. Any flaws in the investigational products will be reported to the sponsors.

Sample label for tested vaccine vials

FOR CLINICAL TRIAL USE ONLY

Recombinant Human Type 5 AdenovirusVector Based Ebola Vaccine(Ad5-EBOV)

Lot Number201411001        Expiration Date2015.10.30

StorageStore at 28°C Away from light. Do not freeze

Beijing Institute of Biotechnology

Tianjin CanSino Biotechnology Inc.

 

Sample label for tested vaccine boxes

FOR CLINICAL TRIAL USE ONLY

Recombinant Human Type 5 AdenovirusVector Based Ebola Vaccine(Ad5-EBOV)

Lot Number201411001        Expiration Date2015.10.30

StorageStore at 28°C Away from light. Do not freeze

Beijing Institute of Biotechnology

Tianjin CanSino Biotechnology Inc.

6.4   Administration

The injection will be given intramuscularly in in the deltoid muscle in participants.

Before injection, 75% alcohol is used for disinfection at the injectionsite, intramuscular vaccination will be administrated several minutes later. Shaking the vaccine before use and make vaccine fully dissolved in sterile water for injection. No intravascular, intradermal or subcutaneous injection is allowed with the investigational vaccine. During the vaccination and the 6-hour safety surveillance after vaccination, appropriate emergency medical equipment and doctors should put on standby in case that possible allergic reaction after injection.

Only the investigator or medically qualified designee will give the injection. For the purpose of vaccine/placebo administration, a medically qualified member of the study staff is a study physician or a study nurse, as applicable to the investigator’s local practice.

6.5   Transportation and Storage

The investigational vaccines should be stored in a safe and locked refrigerator where nobody can get them without authorization. Sponsors should evaluate the storage conditions of the research center before the study to make sure that the vaccines will be stored under appropriate condition during the study.

The vaccine storage temperature should be kept +2°C to +8°C during the transportation from the sponsors to the clinical research center. The temperature should be checked during transportation by suitable temperature monitoringinstrument. When receiving the vaccines at the study center, the receiver should check the vaccines’ number, quality and the maintenance of cold chain during transportation, and fill in the vaccine receipt form.

The temperature should be recorded every day during storage by temperature monitoring instrument and documented each workday manually (once in the morning, once in the afternoon). Frozen indicating instrument should be put near the vaccines to monitor if they are frozen.

7         SAFETY ASSESSMENTS AND ADVERSE EVENT REPORTING

7.1   Adverse event and adverse reaction

An adverse event (AE) is any untoward medical occurrence in a participant administered an investigational product and which does not necessarily have a causal relationship with this treatment. An adverse reaction (AR) is all untoward and unintended responses to a medical product related to any dose administered.

An unexpected adverse reaction is an adverse reaction, the nature or severity of which is not consistent with the applicable product information:

-               Investigator's brochure for an unauthorized experimental product

-               Summary of product characteristics for an authorized product.

For safety assessments, the primary outcome measure is the solicited adverse reactions within 7 days, other outcome measures include all the adverse events within 28 days after vaccination, changes of laboratory measures, and the serious adverse events during the whole study period.

Participants will remain in the clinic to observe for the occurrence of any adverse events for at least 6 hours after receipt of vaccination. For the next 28 days following vaccination, any injection-site adverse events and systematic adverse events will be recorded by the participants on the diary cards. Blood samples collected from participants before the vaccination, on day 3, 14, 28, 56 after vaccination will be tested for significant changes of laboratory testing from baseline.

All adverse events (including laboratory measures) will be graded according to “The standard guidelines for adverse reactions grading of vaccine clinical trials” issued by China state Food and Drug Administration (SFDA).14 The presence of solicited and unsolicited adverse events and any serious adverse events will be described in terms of the incidence, intensity and relation to vaccination. The incidence of adverse events will be based on the most severe response, and expressed in terms of the number and proportion of individuals who had adverse events in each group.

7.2   Grading for adverse events

7.2.1          Injection site

Pain (at the injection site)

Grade 1(mild)= mild pain when the injection site was touched

Grade 2(moderate)= moderate reactions such as withdrawal when the injection site was touched

Grade 3(severe) =crying or refuse to be touched at the injection site

Grade 4(potentially life threatening) = emergency or hospitalization

Mucocutaneous (at the injection site)

Grade 1 (mild) = redness

Grade 2(moderate)= disperse, rash-like tetter, dry, desquamation

Grade 3(severe)= blister-like eruptions, moist, desquamation or ulceration

Grade 4(potentially life threatening)= Peeling dermatitis, or erythema multiforme, or like Stevens-Johnsons-syndrom

Induration (at the injection site)

Grade 1 (mild) = diameter <10 mm

Grade 2 (moderate) = diameter ≥ 10 mm - ≤ 25 mm

Grade 3(severe)= diameter ≥26 mm- ≤ 50 mm

Grade 4(potentially life threatening)= diameter >50 mm

Redness(at the injection site)

Grade 1 (mild) = diameter <10 mm

Grade 2 (moderate) = diameter ≥ 10 mm - ≤ 25 mm

Grade 3(severe)= diameter ≥26 mm- ≤ 50 mm

Grade 4(potentially life threatening)= diameter >50 mm

Swelling (at the injection site)

Grade 1 (mild) = diameter <10 mm and no impairment of movements

Grade 2 (moderate) = diameter ≥ 10 mm - ≤ 25mm or impairment of movements

Grade 3(severe)= diameter ≥26 mm- ≤ 50 mm

Grade 4(potentially life threatening)>50 mm

Exanthema (at the injection site)

Grade 1 (mild) = diameter <10 mm

Grade 2 (moderate) = diameter ≥ 10 mm - ≤ 25 mm

Grade 3(severe)= diameter ≥26 mm- ≤ 50 mm

Grade 4(potentially life threatening)= diameter >50 mm

Itch(at the injection site)

Grade 1 (mild) = slight itch at the injection site

Grade 2 (moderate) = moderate itchon the injection arm

Grade 3(severe)= systemic itch

7.2.2          Systemic reactions

Fever (axillary temperature)

Grade 1(mild) = ≥ 37.1°C - ≤ 37.5°C

Grade 2(moderate) = ≥ 37.6°C - ≤ 39.0°C

Grade 3(severe) =39.0°C

Allergic reactions

Grade 1(mild) = pruritus without rashes

Grade 2(moderate) = localized urticaria

Grade 3(severe) = generalurticaria, angioedema

Grade 4(potentially life threatening)= severe allergic reactions

Headache

Grade 1(mild) = mild symptoms that do not interfere with normal daily activities, need no treatment

Grade 2(moderate) = transient headache, symptoms that have a mild impact on normal daily activities, may need some painkiller

Grade 3(severe) = symptoms that have a significant impact on normal daily activities, and need treatment.

Grade 4(potentially life threatening)= stubborn headache stubborn headache, need hospitalization

Fatigue

Grade 1(mild) =mild symptoms that do not interfere with normal daily activities, last <=48 hours

Grade 2(moderate) = symptoms that have an impact on normal daily activities, last >48 hours

Grade 3 (severe) = symptoms that have an impact on normal daily activities, last >72 hours

Grade 4 (potentially life threatening) =emergency or hospitalization

Vomiting:

Grade 1(mild) = once per 24 hours, do not interfere with normal food intake and no impairment of activities

Grade 2(moderate) = twice to three times per 24 hours, food intake significantly reduced or limitations of activities

Grade 3(severe) = > four to six times per 24 hours, little food intake, necessity of intravenous infusion

Grade 4(potentially life threatening) = >could not eator drink for more than 24hs

Diarrhea:

Grade 1(mild) = slightly or transient, twice to three times of watery stools per day, or continuously slight diarrhea within one week

Grade 2(moderate) = moderate or continuously, four to five times per day or diarrhea, symptom continus more than one week

Grade 3(severe) = > six times of watery stool per day, or bloody stool, postural hypotension, electrolyte imbalance and necessity of intravenous infusion > 2 liter

Grade 4(potentially life threatening) = hospitalization due to hypertensive shock

Muscle pain

Grade 1(mild) = transient, do not interfere with normal daily activities, treatment unnecessary

Grade 2(moderate) = symptoms that have a mild impact on normal daily activities

Grade 3(severe) = serious, symptoms that have a significant impact on normal daily activities, need treatment

Grade 4(potentially life threatening) = muscular necrosis, emergency of hospitalization

Joint pain

Grade 1(mild) = transient, do not interfere with normal daily activities, treatment unnecessary

Grade 2(moderate) = symptoms that have a mild impact on normal daily activities

Grade 3(severe) = serious, symptoms that have a significant impact on normal daily activities, need treatment

Grade 4(potentially life threatening) =emergency of hospitalization

Throat pain

Grade 1(mild) = transient, do not interfere with normal daily activities, treatment unnecessary

Grade 2(moderate) = symptoms that have a mild impact on normal daily activities

Grade 3(severe) = serious, symptoms that have a significant impact on normal daily activities, need treatment

Grade 4(potentially life threatening) =emergency of hospitalization

Cough

Grade 1(mild) = transient, treatment unnecessary

Grade 2(moderate) = continuous coughs, response to treatment

Grade 3(severe) = paroxysmal coughs, treatment uncontrolled

Grade 4(potentially life threatening) = emergency of hospitalization

7.2.3          Safety laboratory measures

Hemoglobin

Grade 1(mild) = 9.5-10.5 gm/dL

Grade 2(moderate) = 8.0-9.4 gm/dL

Grade 3(severe) = 6.5-7.9 gm/dL

Grade 4(potentially life threatening) <6.5 gm/dL

White blood cell count increase

Grade 1(mild) = increase~12.999×109/L

Grade 2(moderate) = 13~14.999×109/L

Grade 3(severe) = 15~30×109/L

Grade 4(potentially life threatening) >30×109/L

White blood cell count decrease

Grade 1(mild) = 2.53.5×109/L

Grade 2(moderate) = 1.52.499×109/L

Grade 3(severe) = 1.01.499×109/L

Grade 4(potentially life threatening)<1.0×109/L

Total lymphocyte count

Grade 1(mild) =0.7~0.799×109/L

Grade 2(moderate) =0.5~0.699×109/L

Grade 3(severe) =0.25~0.499×109/L

Grade 4(potentially life threatening)< 0.25×109/L

Platelets decreased

Grade 1(mild) = 95100×109/L

Grade 2(moderate) =80~ 94.9×109/L

Grade 3(severe) =25.0~79.9×109/L

Grade 4(potentially life threatening)< 25.0×109/L

Prothrombin time

Grade 1(mild) = increase fold 1.01.10 ×14 seconds

Grade 2(moderate) = increase fold 1.111.20×14 seconds

Grade 3(severe) = increase fold 1.211.25×14 seconds

Grade 4(potentially life threatening) >1.25×14 seconds

Activated partial thromboplastin time

Grade 1(mild) = increase fold 1.01.2×40 seconds

Grade 2(moderate) = increase fold 1.211.4×40 seconds

Grade 3(severe)= increase fold 1.411.5×40 seconds

Grade 4(potentially life threatening)= increase fold >1.5×40 seconds

Creatinine

Grade 1(mild) = increase fold 1.11.5×106μmol/L

Grade 2(moderate) = increase fold 1.63.0×106μmol/L

Grade 3(severe) = increase fold 3.16×106μmol/L

Grade 4(potentially life threatening)= increase fold >6×106μmol/L

Alanine transaminase

Grade 1(mild) = increase fold 1.252.5×40U/L

Grade 2(moderate)= increase fold 2.65×40U/L

Grade 3(severe) = increase fold 5.110×40U/L

Grade 4(potentially life threatening)= increase fold>10×40U/L

7.3       Serious adverse event/reaction (SAE)

A serious adverse event/reaction is occurrence of any untoward medical during the whole study period that:

-               Results in death.

-               Is life-threatening (an event in which the participant is at risk of death at the time of the event; it does not refer to an event which hypothetically might have caused death if it was more severe).

-               Results in persistent or significant disability/incapacity.

-               Requires hospitalization or prolongation of an existing hospitalization.

-               Is a congenital anomaly/birth defect.

In addition, medical and scientific judgment will be exercised in deciding whether other conditions will also be considered serious, such as important medical events that may not be immediately life-threatening or result in death or hospitalization but may jeopardize the participant’s safety or may require intervention to prevent one of the other outcomes listed in the definition above. These will also be considered serious. Examples of such events are intensive treatment in an emergency room or at home for allergic bronchospasm; blood dyscrasias or convulsions that do not result in hospitalization; or development of drug dependency or drug abuse.

7.3.1          Reporting SAEs

Any serious adverse event, including death due to any cause, which occurs during this study, whether or not related to the investigational products, must be reported immediately (within 24 hours of the investigator’s knowledge of the event) by telephone or fax to the sponsors, principle investigator, JSCDC IRB, at the following number:

Principal Investigator: Feng-Cai Zhu, Tel: +86-25-83759418, Fax: +86-25-83759409

Representative of sponsor: Wei Chen, Tel: +86-13910789661

JSCDC IRB: Hui-Yuan Cai, Tel: +86-25-83759406; Fax: +86-25-83759406

The unblinding of single cases by investigators in the course of the clinical trial will only be performed if relevant for the safety of the participant.

The report for CRO will containavailable information concerning the serious adverse event to enable CRO to file a report which satisfies regulatory reporting requirements. This will be done on the SFDA SAE-form. This will be a preliminary report. The final report will be provided after evaluation by the sponsor. according to ICH standard timelines. In addition to the initial 24-hour report, a completed, separate SAE report is to be sent to CRO via fax or mail within 48 hours of the event. All SAEs will be recorded on the case report form and source documents.

7.3.2          Expedited reporting of SUSARs to regulatory authorities and investigators

All suspected adverse reactions related to an investigational products, that are both unexpected and serious (SUSARs) are participant to expedited reporting. Also post-study SUSARs that occur after the patient has completed a clinical trial and are reported by an investigator to the sponsor, qualify for expedited reporting.

The sponsor is responsible for the prompt notification to all concerned investigators, the JSCDC IRB and competent authority of all relevant safety information previously described.

Fatal or life-threatening SUSARs will be notified by the sponsor. to the competent authority and the JSCDC IRB as soon as possible but no later than seven calendar days after the sponsor has first knowledge of the minimum criteria for expedited reporting.

In each case relevant follow-up information will be sought and a report completed as soon as possible. It will be communicated to the competent authority and the JSCDC IRB within an additional eight calendar days.

All other SUSARs and safety issues deserving expedited reporting must be reported to the competent authority and dependent on national provisions to the JSCDC IRB in the concerned countries as soon as possible but no later than 15 calendar days after the sponsor has first knowledge of the minimum criteria for expedited reporting.

Expedited reporting is not usually required for reactions which are serious but expected, or for non-serious adverse reactions whether expected or not.

7.4       Safety data monitoring by DSMB

A five-member external DSMB has been built for the safety data monitoring of this trial. The composition of DSMB including a biostatistician, an epidemiologist, an expert in anti-infection, an expert in haematology, and an ethicist (detail information about the DSMB see Constitution of Data and Safety Monitoring Board).

The DSMB will blindly review safety continuously during the trial and will advise on progression of the trial. It will review the safety data from the trial as generated by the investigators, in order to advise whether or not the study integrity remains intact and whether or not there are any safety concerns worth early notice. DSMB will review the reported safety data in the participants for the first 7 days after vaccination. During the study period, if an increase of risk for participants is noticed, the DSMB should promptly inform the principle investigator and sponsors. Sponsors, investigators and DSMB will have a panel meeting, and then DSMB will make final decision to pause or all an early termination of the study.

The following information should be provided to the DSMB:

-      All subsequent protocol amendments, informed screening or study consent form changes or revisions of other documents originally submitted for review.

-      All adverse events reported during the first 7 days after vaccination

-      Grade 3 adverse experiences occurring during the vaccination period (from day 0 to day 28), regardless of relationship to the study vaccine.

-      Serious adverse events (SAEs) occurring during the study, regardless of relationship to the study vaccine.

-      New information that may affect adversely the safety of the participants or the conduct of the study.

-      All the information related to SAEs will be reported to DSMB, and the DSMB are responsible to give a final decision of causality for SAEs.

7.5       Withdrawal due to adverse events (see also section on participant withdrawal)

Withdrawal due to AE should be distinguished from withdrawal due to other reasons unrelated to safety issues, according to the definition of AE noted earlier and recorded on the appropriate AE CRF page.

When a participant withdraws due to an SAE, the SAE must be reported in accordance with the reporting requirements defined above.

8         LABORATORY ASSAY AND IMMUNOGENICITY ASSESSMENTS

8.1       Antigen-specific antibody responses

Serum antibody titers against the Zaire Ebola virus GP will be determined at day 0 (immediately before vaccination), day 3, day 7, day 14, day 28, day 56, day 112, and day 168. The primary time point of antibody measurement is day 28. The antibody responses in different time points will be compared between vaccine group and placebo group. ELISA will be applied for the detection of serum antibody against Zaire Ebola virus GP.11,15

8.2       Antigen-specific CD4+, CD8+ T cell responses

Peripheral blood mononuclear cells (PBMC) will be isolated from venous blood and stimulated withoverlapping peptide pools of GP protein.The antigen-specific CD4+ and CD8+ T cell responses will be tested at day 0, day 7, day 14, day 28, day 56, day 112, day 168. Intracellular cytokine staining (ICS) assay will be applied to detect the IL-2,INF-γ, and TNF express by T cells after the specific peptides stimulation.16,17

Enzyme-linked immunospot (ELISpot) assay18 will be used to detection in human T cell immune response who injected Ebola virus vaccine base on the Ebola virus envelope glycoprotein (EBOV-GP) at day 0, 14 and 28 (primary time point of antibody measurement) and day 168.

8.3       Neutralizing antibody titers response toAd5

Neutralizing antibody titers response to human Ad5 will be detected at day 0, day 3, day 7, day 14, day 28, day 56, day 112, day 168. Ad5-specific neutralizing antibody testing will be detected by using the replication defective human type 5 adenovirus expressing luciferase (Ad5-luc).9 Ad5-specific neutralizing antibody in serum could bind to Ad5-luc to inhibit the expression of luciferase in cells. So, the monolayer A549 cells, which have been infected by the mixture of quantitative Ad5-luc and equivalent serum with different dilution, are cultured for 24 hours, and the neutralizing ability of different serums could be estimated by testing the activity of luciferase in cells.19

The neutralizing antibody titers against human Ad5 pre-vaccination and the post-vaccination will be compared to reveal the difference. The relationship between the pre-existing antibody titers against Ad5 and the specific immune response against Zaire Ebola virus (including both humoral and cellular responses) will be explored.

9.         DATA COLLECTION AND MANAGEMENT

9.1       Source documents and source data

The purpose of source documents is to document the existence of the participant and substantiate the integrity of the trial data collected. The Investigator must maintain the trial source documents accurate, complete, legible and up to date.

Examples of source documents are: participant screening, laboratory measure reports, enrolment log, participant’s diary cards, hospital records, informed consent forms, investigational dispensing and reconciliation forms, participant’s file and records kept at the pharmacy or at the laboratories, mail, certified letters.

Source data are the data contained in source documents (originals or certified copies). The investigator is responsible for the accuracy and completeness of the data reported in source documents. Data reported in the CRFs that are derived from source documents should be consistent with source documents and any discrepancies should be explained.

All CRFs must be signed by the Investigator. Incorrect data must be crossed-out with a single line, then initialed and dated. Correction fluid or similar corrective methods that mask the original data will not to be used. These rules also apply to the completion of SAE Reporting Forms, Data Correction Forms, and ICFs.

9.2       Clinical data management

The investigators should fill in all case report forms (CRF). CRF is used to record data in clinical trials, is an important part of clinical trials and research reports, filling shall be clear and intact, and also should be completed with Chinese language and black pen. Only authorized investigators could correct the errors in the CRFs, The original record can’t be obliterated or overwrite. Investigator should draw a horizontal line across the original data which should be corrected, and specify the corrected data in the space next to them, and noted the signature and date.

According to the project requirements, the data collection, biological sample collection and examination should be done in the visit window, the original documents and records shall be complete, result of the examination also should be timely entered into case report form(CRF).

Auditors should conduct regular and irregular audits of data records until CRF are completed, auditors should carefully verify CRF number of the participants, the number of pages in each CRF and necessary signatures of researchers. The main contents of audits should be focused on signed informed consent, volunteer screening into the group, vaccination, management of the investigational vaccine, safety observation and immunogenicity of specimen collection and preservation, Consistency between research data and the original data is the emphasis of audits. Manual verification results shall be recorded. Transfer of CRF research data should be documented. For each batch of data, double entry, quality control and triggers to computerized logic and/or consistency checks will be systematically applied in order to detect errors or omissions. Queries will be generated and submitted through Data Clarification Forms to the Investigator for resolution.

9.3       SAE data management

During the trial, all the SAEs will be reported on SAE Reporting Forms required by SFDA and integrated into the Sponsor’s SAE database. Upon receipt of an SAE Reporting Form, the data will be entered into the database after a duplicate check. Each SAE is assigned a case identification number. Entered data will be independently verified against the original SAE forms. All SAEs are then reviewed by the DSMB. Each SAE is reviewed, locked and approved in the database. Any follow-up information concerning a locked and approved SAE will be incorporated and a new version of the SAE will be created.

9.4       Archiving

Following closure of the study, the investigator must maintain all site study records in a safe and secure location. The records must be maintained to allow easy and timely retrieval, when needed (e.g., audit or inspection), and, whenever feasible, to allow any subsequent review of data in conjunction with assessment of the facility, supporting systems, and staff. Where permitted by applicable laws/regulations or institutional policy, some or all of these records can be maintained in a validated format other than hard copy; however, caution needs to be exercised before such action is taken. The investigator must assure that all reproductions are legible and are a true and accurate copy of the original ones, and meet accessibility and retrieval standards, including re-generating a hard copy, if required. Furthermore, the investigator must ensure there is an acceptable back-up of these reproductions and that an acceptable quality control process exists for making these reproductions.

Trial-related documents will be maintained for a period of 10 years after final marketing approval of the vaccine, or 10 years after the formal discontinuation of clinical development of the product per the requirements by SFDA and IRB/IEC. The site investigators must be aware of all requirements and retain protocol records in accordance with the longest requirement that pertains to the study. No study document should be destroyed without prior written agreement between the sponsors and the principal investigator. Storage of all trial-related documents will be such that confidentiality will be strictly maintained.

9.5       Database creation and data entry

The person, who is responsible for data management, establish the database structure and inspection procedures to ensure that the database can be correctly converted to SAS file format, and the database structure can also be modified and confirmed by tentative data entry.

Before CRF entry, data management staff will carry out the CRF verification again, mainly to see if there are obvious errors and omissions. Data entry staff will start data entry after several training, double entry will be accepted.

The consistency check of the database that be independently completed by two persons will be executed. Inconsistent values and information should be reported. After that, the raw data will be checked one by one and the mistake will be corrected until database is left in a consistent state. The computer program that has been prepared and confirmed will be applied to do logical consistency check. Before modifying the database, Query table should be shown and confirmed by researchers until there is no question. A certain proportion of CRF will be randomly selected to finish quality control, and to compare with the data in the database, to ensure that the data in the database and the CRF content are consistent.

9.6       The database lock

Blind review of the database is required before Statistical analysis. The aim of blind review is to determine the population that will be analyzed according the evaluation criteria, including full analysis set (FAS) under the Principle of ITT (intention to treat analysis) , per-protocol set (PPS) and safety analyzes data set, confirmation of the deviation from the project and other influences on database. Database will be locked after blind review be confirmed.

10.     STATISTICS PLAN AND STATISTICAL ANALYSIS

10.1    Interim analysis

Interim analyses will be conducted by the independent statistical party after the last participant completes Visit 5 on day 28, and all the safety data and immunogenicity data up to day 28 will be collected. The data will be unblinded by representatives of statistical party, sponsor, clinical research investigator and CRO. The un-blinding data will not be confidential and therefore the clinical trial site will not be kept in blind. The results will not influence the conduct of the trial in terms of early termination or later safety or immunogenicity endpoint assessments. After the review of DSMB, the interim analysis report could be submitted to the investigator and the sponsor.

10.2    The final analysis

After the last participant complete the Visit 8 (168 days after vaccination), database entry is completed, blind review is finished and database is locked, the final analysis will be done by the independent statistical party.

10.3    Analyzed data sets definition

10.3.1      Data set for safety evaluation

All randomized participants who received vaccination should be included in the safety evaluation. Thus, the safety analysis will be performed on the basis of (Intention-To-Treat) ITT cohort.

Events will be reported on per-individual basis, i.e. counting individuals rather than events. This means that even if a participant suffered a same event repeatedly during the follow-up, the event will be counted only once, except for SAEs. Repeated same adverse events in participant will be summarized according to the following rule: if a participant suffered the same adverse event more than once, the event will be assigned the worst severity, the closest relationship to the vaccination and the earliest starting date. In the listings, however, all occurrences of the adverse events will be shown.

10.3.2      Data set for immunogenicity evaluation

Full analysis set (FAS) for immunogenicity analysis: FAS is based on ITT (intention to treat analysis) principle to determine the participants, All of the participants that meet the inclusion/exclusion criteria, randomization, receiving vaccination, and have at least one blood testing result after vaccination, were included in the FAS set for immunogenicity.

Per-protocol set (PPS): It is a subset of FAS. In this set, all participants that meet the inclusion / exclusion criteria and complete the vaccine inoculation within visit window according to the protocol, and complete the blood collection on day 0 and 28, with no significant deviation or violation of protocol.

In this study, the FAS are the primary analysis set for immunogenicity evaluation, but the PPS will also be analyzed at the same time. Any difference of analysis results existed between PPS and FAS, will be discussed in the report.

10.3.3      Statistical methods

Safety analysis of this experiment is mainly descriptive analysis of incidence rate of adverse reaction or adverse events. A chi-square test can be used to compare the proportion of groups, Fisher's exact test will be used when it is necessary. Analysis of immunogenicity indicators on antibody levels need to do logarithmic transformation, the results of analysis should be shown in GMT, standard deviation, median, minimum and maximum values and 95% confidence intervals, chi-square test can be used to compare categorical indicators between groups such as positive conversion rate of immune response, if it is necessary, Fisher's exact test will be used. Statistical analysis method of repeated measures data can be used to analyze experimental data at different time points in this study.

SAS (version 9.3) was used for all analyses, test statistics and the corresponding p values are given. all statistical tests were two-sided and significance was set at P≤0.05 (more detailed information, please read the reference, the interim-term statistical analysis plan and the final statistical analysis plan).

11.    ETHICAL AND LEGAL ISSUES

11.1    Guideline

The study will be conducted according to Good Clinical Practice (GCP), the Declaration of Helsinki, and local rules and regulations of China. Submission of the protocol and any protocol amendments to regulatory agencies will occur in accordance with local regulatory requirements. When submission to the local regulatory authority is required, the timing of the submission relative to IRB submission or approval and whether or not the authority will provide their approval of or favorable opinion on the protocol or amendment before it can be implemented will depend on local regulatory requirements.

11.2    Institutional Review Board

The investigator is responsible for obtaining written approval for the clinical study protocol (including all substantial protocol amendments), the written participant informed consent form, informed consent updates, participant recruitment procedures (e.g. advertisements) and any other written information to be provided to participants from an IRB which complies with local regulatory requirements. Any amendments will require approval by the IRB.

The only circumstance in which an amendment may be initiated prior to JSCDC IRB approval is where the change is necessary to eliminate apparent immediate hazards to the participants. In that event, the investigator must notify the Institutional Review Board and the sponsor in writing immediately after the implementation.

A final study notification will be forwarded by the investigator to the JSCDC IRB within 90 days after the study has been completed or in the event of premature termination of the study within 15 days. Copies of all clinical study status reports (including termination) will be provided by an investigator to CRO.

11.3    Ethical Conduct of the Study

- To ensure that he/she has sufficient time to conduct and complete the study and has adequate staff and appropriate facilities and equipment which are available for the duration of the study and to ensure that other studies do not divert essential participants or facilities away from the study at hand.

- To submit an up-to-date curriculum vitae or Investigator Biography and other credentials to the sponsors and where required to relevant authorities. It is recommended that this documentation indicates any previous clinical research experience and history of training in GCP.

- To acquire the reference ranges for laboratory tests performed locally and, if required by local regulations, obtain the laboratory’s current certification or Quality Assurance procedure manual.

- To ensure that all back-up clinical samples (including serum samples) are retained onsite according to the approval of the sponsors.

- To perform no other biological assays on the clinical samples except those described in the protocol or its amendment(s).

- To prepare and maintain adequate source data or raw data designed to record observations, and other data pertinent to the study.

- To conduct the study in compliance with the protocol any amendment and “Good Clinical Practice” (GCP) and all applicable regulatory requirements.

- To cooperate with a representative of the sponsors in the monitoring process of the study and in resolution of queries about the data.

- To permit drug regulatory agencies and sponsors’ audits.

11.4    Protocol Amendments and Administrative changes

No changes to the study protocol will be allowed unless approved by sponsors. This does not apply to changes made to reduce discomfort or avert risk to study participants. Furthermore, in the event of a medical emergency, the investigators shall perform any medical procedures that are deemed medically appropriate. The principle investigator must notify the sponsor of all such occurrences.

Written IRB approval of protocol amendments is required prior to implementation, except where permitted by all applicable regulatory requirements; administrative changes and amendments not submitted for approval are submitted to IRB for information only. Any amendment/administrative change to the protocol will be adhered to by the participating centre(s) and will apply to all participants. Submission of protocol amendments to regulatory agencies will occur in accordance with local regulatory requirements. When submission to the local regulatory authority is required, the timing of the submission relative to IRB submission or approval and whether or not the authority will provide their approval of or favorable opinion on the amendment before it can be implemented will depend on local regulatory requirements.

11.5    Confidentiality of Data and Access to Participant Records

Prior to initiation of the trial, the investigators will sign a fully executed confidentiality agreement with the sponsor. All study-related information will be stored securely at the study sites. All participant information will be stored in locked file cabinets in areas with access limited to study staff. All laboratory specimens, reports, study data collection, process, and administrative forms will be identified by coded number only to maintain participant confidentiality. All computer entries will be done by coded numbers only, and all local databases will be secured with password-protected access systems. Forms, lists, logbooks, appointment books, and any other listings that link participant ID numbers to other identifying information will be stored in a separate, locked file in an area with limited access.

Sponsor personnel, the IRB and the regulatory authorities will have direct access to source data/documents.

11.6    Benefits and risks

Participants will not pay for vaccination or physical examination in this clinical trial. Meanwhile, they can get reasonable compensation for transportation costs, time costs, blood donation and nutritional costs. Participants will receive vaccination of the Ad5-EBOV or placebo. Immunization with the Ad5-EBOV may protect participants from the threat of Ebola virus, but we can’t guarantee that. In this study, participants may suffer some adverse reactions, such as fever, pain at injection-site, swelling, etc. Common adverse reactions will generally resolve in 3-5 days. Previous adenovirus vaccines in other countries had reported that adenovirus as vectors may cause an increase in bleeding time in a short period of time. But it won’t last and generally is not life-threatening. Thus, several adenoviral vector vaccines have been approved for market in foreign countries. In addition, clinical research of VSV vector vaccine in Canada has found that vaccination may cause joint pain. So we add joint pain as a solicited adverse reaction in this study. Previous studies have indicated that immunization with Ad5 vector based or other viral vector based vaccines may increase the risk of HIV-1 acquisition,20 so we suggest all vaccine recipients avoid HIV associated high-risk behavior such as dangerous sexual behavior, intravenous drug use, or illegal blood transfusion.

If participants do not want to get inoculated against the Ad5-EBOV, there is no alternative because no Ebola vaccine is available in China currently.

12.    FINANCIAL CONTRACT AND INSURANCE COVERAGE

An agreement will be signed by all the parties involved in the trial’s performance, if relevant. Adequate insurance coverage for all participants to be included in the trial is supplied by the Sponsor.

13.    PUBLICATION OF STUDY RESULTS

Sponsors have no objection to publication by Investigator of any information collected or generated by Investigator, whether or not the results are favorable to the Investigational Product. However, to ensure against inadvertent disclosure of Confidential Information or unprotected Inventions, Investigator will provide sponsors an opportunity to review any proposed publication or other type of disclosure before it is submitted or otherwise disclosed.


 

14.    APPENDIX

Appendix 1 Informed consent form for the participants in low dose group (Version1.2)

Informed Consent Form (ICF) for the Participants in the Low Dose Group

 

A Phase 1 Double-blind, Dose-escalation, Placebo-controlled Clinical Trial to Evaluate the Safety, Tolerability and Immunogenicity of the Recombinant Adenovirus Type 5 Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults aged between 18 and 60 years in China.

The clinical trial has been approved for clinical trials in December 12, 2014 (approval number: 2014JTL005).

Principal Investigator: Feng-Cai ZhuTel: +86-25-83759418

Organization: Jiangsu Provincial Center for Disease Control and Prevention

Sponsor: Beijing Institute of Biotechnology

Tianjin CanSino Biotechnology Inc.

Protocol Number& Version: JSVCT020 (Version 1.2)

 

This Informed Consent Form has two parts:

·           Information Sheet (to share information about the study with you)

·           Certificate of Consent (for signatures if you agree to participate)

You will be given a copy of the full Informed Consent Form

 

1. Brief introduction

I am going to give you information and invite you participate in this study.

Before you make a decision whether or not to take part in this study, this Informed Consent Form will help you understand the details of the entire study, the risks and benefits of participants as well as what needs to be done. There may be some words that you do not understand. Please ask me to stop as we go through the information and I will take time to explain. Please carefully read the following information. If you have any question later, you can consult me or the study doctor any time.

 

2. Objectives

Ebola viruses (EBOVs) are enveloped, non-segmented, negative-strand RNA viruses belonging to the family Filoviridae. They are known to cause lethal hemorrhagic fever in humans and non-human primates with a mortality rate up to 90%. The current outbreak in West Africa, (first cases notified in March 2014), is the largest and most complex Ebola outbreak since the Ebola virus was first discovered in 1976. On August 8, the WHO Director-General declared this outbreak a Public Health Emergency of International Concern.

This is a phase 1 clinical trial. We are going to evaluate the safety, tolerability and immunogenicity (including humoral and cellular response) of the Ad5-EBOV in healthy adults.

 

3. Intervention

The candidate Recombinant Adenovirus Type 5 Vector Based Ebola Vaccine (Ad5-EBOV) against Ebola disease is developed by Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc. The Zaire Ebola virus in 2014 causing the most serious outbreak was considered to be a new epidemic strain, with GP homology of the gene was only 97.6%, compared to the GP gene of the strain in 1976. All the other Ebola Vaccines undergoing the clinical trial were developed based on the Zaire-Mayinga (1976 strain). This experimental Ad5-EBOV is the first Ebola vaccine developed according to the 2014 epidemic strain.

 

4. Participants

A total of 120 participants will be enrolled in this study. Of them, first 60 participants will be allocated to the low dose group, and randomly allocated to receive the low dose vaccine or placebo in a ratio of 2:1. After we confirm that the safety of the low dose group during the first seven days is acceptable, then another 60 participants will be recruited for the high dose group and randomly allocated to receive high dose vaccine or placebo in a ratio of 2:1.

We are now recruiting the participants for the low dose group.

 

5. Information on the Investigational Products

Experimental vaccine Ad5-EBOV, developed by Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc., is a replication defective Adenovirus Type 5 Vector based vaccine which expresses Ebola virus Zaire (Guinea, 2014) envelope glycoprotein. The final product is lyophilized white powder, with 4.0×1010vp/vial. The antigen contents in the Ad5-EBOV were measured according Ebola vaccine antigen standard by National Institute for Food and Drug Control (NIFDC).

The placebo is lyophilized-dried powder, contains same formulation of excipients as vaccine, with no virus particle which could express Ebola virus Zaire envelope glycoprotein. The placebo wasalso measured to be qualified by NIFDC.

Low dose group:

Experimental: injection with 4×1010vp/1ml Ad5-EBOV; one vial of vaccine dissolved (4.0×1010vp/vial) in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

Control: one shot of 0vp/1ml placebo; dissolved one vial placebo in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

High dose group:

Experimental: 1.6×1011vp/2ml Ad5-EBOV; two vials of vaccine dissolved (4.0×1010vp/vial) in 1ml sterile water for injection. Double shots of 0.8×1011vp/dose will be allocated intramuscularly in two arms at day 0 (one shot in each arm).Thus, participants in the high dose group will be vaccinated with a dose of 1.6×1011vp (=4.0×1010vp/vial× 4).

Control: double shots of 0vp/1 ml placebo; two vials of placebo dissolved in 1ml sterile water for injection. Two injections will be allocated intramuscularly in two arms at day 0 (one shot in each arm).

 

6. Content and procedures of the study

The study included three phases: Screening, vaccination, and follow-up. If you agree to participate in this study, the doctor will give you a medical examination and you need to donate about 13ml blood for laboratory tests (including routine blood test, blood biochemical test, active partial thromboplastin time, prothrombin time and HIV test) to confirm that you are suitable to enter this study according to the inclusion criteria and the exclusion criteria. If you are not eligible for this study you will be excluded from this study before the treatment allocation.

We are now recruiting the participants for the low dose group.If you meet all the inclusion criteria and did not meet any of the exclusion criteria, you will be allocated randomly to receive the low dose vaccine or placebo. Youwill have 2/3 chance of receiving the low dose vaccine. The immunization schedule is one shot at day 0. You will be asked to stay in the clinic for at least 6 hours for safety observation. While you are in the clinic, the doctor will show you how to record temperature and adverse events on diary card.

You will getadiary card, athermometer, and a measuring device to take home. The doctor will also show you how to measure your temperature at home. You will be asked to record any injection-site adverse events and systematic adverse events on the diary cards for the following 28 days.

You are required to return to clinic at day 3, day 7, day 14, day 28, day 56, day 112, day 168 after vaccination for the collection of safety data and blood taking for the tests of humoral and cellular immune responses. The laboratory tests will be retested between regular visits to evaluate a change in your health. Besides, your blood will also be used for some genetic testing such as HLA typing, but all the results will be kept confidential and only used for research purposes only. You are asked to donate about 50 ml blood at each visit. The detailed information about the volume of blood taking is listed as follows.

Visit

V0

V1

V2

V3

V4

V5

V6

V7

V8

Daily Volume (ml)

13

55

45

45

52

62

52

42

52

Cumulative Volume (ml)

13

68

113

158

210

272

324

366

418

You will be asked to donate a maximum of 418 mlof blood during the whole study. The volume of blood taking will not do harm to a healthy adults.

 

7. Duration

You will attend a total of 9 visits during the follow-up period of 6 months, including the screening visit before vaccination and 8 site visits after vaccination. Additional visits maybe required if you have any worries or concerns about your health.

 

8. Discomfort, adverse reaction and possible risk

By participating in this study it is possible that you may experience some discomfort. The vaccination may cause mild/moderate fever, injection-site pain, redness, soreness, itchiness, or other discomfort. Allergic reactions may be observed occasionally. Generally, these reactions can disappear within 24 hours without treatment. There is a very small chance of infection.

The blood taking may also be accompanied by some risk. Pain, bleeding, bruising, or feeling lightheaded or mild pain may occurred. Syncope or infection at the blood drawing site israre.

Unintended release of some test results such as HLA type could be used by insurers or employers, which may suggest risk of disease for you or your family. Therefore the genetic test results are confidential, we are not going to release the results to you or any unauthorized person.

While the possibility of serious adverse reaction happening due to participating is very low, you should still be aware of the possibility. If something unexpected happens and harm does occur, we will provide youappropriate treatment and corresponding economic compensation.

Previous studies have indicated that immunization with Ad5 vector based or other viral vector based vaccines may increase the risk of HIV-1 acquisition, so we suggest you avoid HIV associated high-risk behavior such as dangerous sexual behavior, intravenous drug use, or illegal blood transfusion.

9. Possible benefits for taking part in this study

All the laboratory examination and vaccinations in study are free. By taking part in this trial, you may develop the immune response to the Ebola virus due to the vaccinations. But the vaccination cannot provide 100% guarantee for resistance against Ebola virus disease. You may not get other healthy benefit for taking part in this study, but your contribution may benefit other people and the whole nation.

10. Reimbursements

In order to compensate your travel expenses, blood denotation and the time lost for participating in this study, you will get a reimbursements of 500 RMB after each visit, and after complete all the 8 visits you will get a total of 4000 RMB as reimbursements. Relevant medical expenses for you to take part in this study are free, such as expense of vaccine, laboratory examination, and so on. You will not get other payments unrelated to the trial.

 

11. Confidentiality of the study data

The results of this study will be given to the sponsor. Your personal information will be kept confidentially. Your name will not appear in any published information on the study or report. In addition, the representative of the China State Food and Drug Administration (SFDA) may examine the medical history record related to this study, so as to verify the accuracy of the data collected in this study, but it will not involve your detailed personal information.

 

12. Sharing of the results

Confidential information will not be shared.The test result in this study that we get will not be shared with you, unless the results indicatethere is a potential safety risk and you need to know. But if you want, you can require the doctors to give you the results.We will publish the results in order that other interested people may learn from our research.The identity of subjects remains confidentialto protect your privacy.

 

13. Right to Refuse or Withdraw

You may choose not to participate.You can change your mind and withdraw (drop out) later.There will be no penalty, and you will not lose any benefits they receive now or have a right to receive proper treatment.

We will tell you if we learn new information that could change your mind about taking part or continuing in this research study. If you want to drop out, you should tell us. We will make sure you can end the study in the safest way.

The study doctor or the study sponsor may decide to let you out of the study without your agreement if, for example:

You do not follow the directions of the study team, or;

The study doctor decides that the study is not in your best interest, or;

The study is stopped by the study sponsor, the institutional review board (IRB), or by a regulatory agency.

The decision of the participation in the study is entirely voluntary. You may withdraw from the study without giving any reason at any time before the study or after it starts. If you withdraw the study for any reason, biological samples (for example, blood samples) that have been collected from you (but not yet fully analyzed) can be destroyed by making a request to the study doctor. However, any data already generated from your samples will be kept to preserve the value of the study. If you leave the study for any reason, you will not need to donate blood or receive any further tests.

 

14. Alternatives to participating

Because there is no Ebola vaccines available in the market at present. If you do not wish you to take part in this study, no other intervention can provide to you. Since there is no Ebola infected patient has been found in China, you may not have the risk of exposure if you stay in China. You can also take some nonspecific precautionary measures such as hand-washing to protect yourself.

 

15. Contact

If you have any questions you may ask the investigator or the study staff now or later, even after the study has started. If you wish to ask questions later, you may contact any of the following:

Feng-Cai Zhu Tel: +86-25-83759418

 

This proposal has been reviewed and approved by the IRB of Jiangsu Provincial Center for Disease Prevention and Control, which is a committee whose task is to make sure that research participants are protected from harm. If you wish to find about more about the IRB, contact:

Hui-Yuan Cai Tel: +86-25-83759406

 

16. Certificate of Consent

By signing this Informed Consent Form, you confirm that you have read this information and have sufficiently considered it, that the doctor has explained this study to you, that your questions have been answered, that you have had enough time to make the decision on the participation, and that you agree to take part in this study. You have learned that you may withdraw from this study at any time without any reason, and the withdrawal from the study will not affect the present or future treatment of you.

 

Name of participant: ____( completed by doctor)

Signature of participant:__ ______       Date:_____ _____ _____( completed by participant)

                                        year/month/day

 

17. Statement of investigator

I confirm that I have explained the content, procedures, possible risks and benefits of this study to the participant, and have given him or her adequate answers about any question, and the family members of the participant have got satisfactory answers.

 

Signature of doctor:_____                                 Date:____ ____ ____

year/month/day

 


 

Appendix 2 Informed consent form for the participants in high dose group (Version1.2)

Informed Consent Form (ICF) for the Participants in the High Dose Group

 

A Phase 1 Double-blind, Dose-escalation, Placebo-controlled Clinical Trial to Evaluate the Safety, Tolerability and Immunogenicity of the Recombinant Adenovirus Type 5 Vector Based Ebola Vaccine (Ad5-EBOV) in Healthy Adults aged between 18 and 60 years in China.

The clinical trial has been approved for clinical trials in December 12, 2014 (approval number: 2014JTL005).

Principal Investigator: Feng-Cai ZhuTel: +86-25-83759418

Organization: Jiangsu Provincial Center for Disease Control and Prevention

Sponsor: Beijing Institute of Biotechnology

Tianjin CanSino Biotechnology Inc.

Protocol Number& Version: JSVCT020 (Version 1.2)

 

This Informed Consent Form has two parts:

·           Information Sheet (to share information about the study with you)

·           Certificate of Consent (for signatures if you agree to participate)

You will be given a copy of the full Informed Consent Form

 

1. Brief introduction

I am going to give you information and invite you participate in this study.

Before you make a decision whether or not to take part in this study, this Informed Consent Form will help you understand the details of the entire study, the risks and benefits of participants as well as what needs to be done. Theremay be some words that you do not understand. Please ask me to stop as we go through the information and I will take time to explain.Please carefully read the following information. If you have any question later, you can consult me or the study doctor any time.

 

2. Objectives

Ebola viruses (EBOVs) are enveloped, non-segmented, negative-strand RNA viruses belonging to the family Filoviridae. They are known to cause lethal hemorrhagic fever in humans and non-human primates with a mortality rate up to 90%. The current outbreak in west Africa, (first cases notified in March 2014), is the largest and most complex Ebola outbreak since the Ebola virus was first discovered in 1976. On August 8, the WHO Director-General declared this outbreak a Public Health Emergency of International Concern.

This is a phase 1 clinical trial.We are going to evaluate the safety, tolerability and immunogenicity (including humoral and cellular response) of the Ad5-EBOV in healthy adults.

 

3. Intervention

The candidate Recombinant Adenovirus Type 5 Vector Based Ebola Vaccine (Ad5-EBOV) against Ebola disease is developed by Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc. The Zaire Ebola virus in 2014 causing the most serious outbreak was considered to be a new epidemic strain, with GP homology of the gene was only 97.6%, compared to the GP gene of the strain in 1976. All the other Ebola Vaccines undergoing the clinical trial were developed based on the Zaire-Mayinga (1976 strain). This experimental Ad5-EBOV is the first Ebola vaccine developed according to the 2014 epidemic strain.

 

4. Participants

A total of 120 participants will be enrolled in this study. Of them, first 60 participants will be allocated to the low dose group, and randomly allocated to receive the low dose vaccine or placebo in a ratio of 2:1. After we confirm that the safety of the low dose group during the first seven days is acceptable, then another 60 participants will be recruited for the high dose group and randomly allocated to receive high dose vaccine or placebo in a ratio of 2:1.

We are now recruiting the participants for the high dose group.

 

5. Information on the Investigational Products

Experimental vaccine Ad5-EBOV, developed by Beijing Institute of Biotechnology and Tianjin CanSino Biotechnology Inc., is a replication defective Adenovirus Type 5 Vector based vaccine which expresses Ebola virus Zaire (Guinea, 2014) envelope glycoprotein. The final product is lyophilized white powder, with 4.0×1010vp/vial. The antigen contents in the Ad5-EBOV were measured according Ebola vaccine antigen standard by National Institute for Food and Drug Control (NIFDC).

The placebo is lyophilized-dried powder, contains same formulation of excipients as vaccine, with no virus particle which could express Ebola virus Zaire envelope glycoprotein. The placebo wasalso measured to be qualified by NIFDC.

Low dose group:

Experimental: injection with 4×1010vp/1ml Ad5-EBOV; one vial ofvaccinedissolved(4.0×1010vp/vial) in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

Control: one shot of 0vp/1ml placebo; dissolved one vial placebo in 1ml sterile water for injection. One injection will be allocated intramuscularly in the arm at day 0.

High dose group:

Experimental: 1.6×1011vp/2ml Ad5-EBOV; two vials of vaccine dissolved (4.0×1010vp/vial) in 1ml sterile water for injection. Double shots of 0.8×1011vp/dose will be allocated intramuscularly in two arms at day 0 (one shot in each arm). Thus, participants in the high dose group will be vaccinated with a dose of 1.6×1011vp (=4.0×1010vp/vial× 4).

Control: double shots of 0vp/1 ml placebo; two vials of placebo dissolved in 1ml sterile water for injection. Two injections will be allocated intramuscularly in two arms at day 0 (one shot in each arm).

 

6. Content and procedures of the study

The study included three phases: Screening, vaccination, and follow-up. If you agree to participate in this study, the doctor will give you a medical examination and you need to donate about 13ml blood for laboratory tests (including routine blood test, blood biochemical test, active partial thromboplastin time, prothrombin time and HIV test) to confirm that you are suitable to enter this study according to the inclusion criteria and the exclusion criteria. If you are not eligible for this study you will be excluded from this study before the treatment allocation.

We are now recruiting the participants for the high dose group. If you meet all the inclusion criteria and did not meet any of the exclusion criteria, you will be allocated randomly to receive the high dose vaccine or placebo. You will have 2/3 chance of receiving the high dose vaccine. The immunization schedule is double shots at day 0. You will be asked to stay in the clinic for at least 6 hours for safety observation. While you are in the clinic, the doctor will show you how to record temperature and adverse events on diary card.

You will get a diary card, a thermometer, and a measuring device to take home. The doctor will also show you how to measure your temperature at home.You will be asked to record any injection-site adverse events and systematic adverse eventson the diary cards for the following 28 days.

You are required to return to clinic at day 3, day 7, day 14, day 28, day 56, day 112, day 168 after vaccination for the collection of safety data and blood taking for the tests of humoral and cellular immune responses.The laboratory tests will be retested between regular visits toevaluate a change in your health. Besides, your blood will also be used for some genetic testing such as HLA typing, but all the results will be kept confidential and only used for research purposes only. You are asked to donate about 50 ml blood at each visit. The detailed information about the volume of blood taking is listed as follows.

Visit

V0

V1

V2

V3

V4

V5

V6

V7

V8

Daily Volume (ml)

13

55

45

45

52

62

52

42

52

Cumulative Volume (ml)

13

68

113

158

210

272

324

366

418

You will be asked to donate a maximum of 418 ml of blood during the whole study. The volume of blood taking will not do harm to a healthy adults.

 

7. Duration

You will attend a total of 9 visits during the follow-up period of 6 months, including the screening visit before the vaccination and 8 site visits after vaccination. Additional visits maybe required if you have any worries or concerns about your health.

 

8. Discomfort, adverse reaction and possible risk

By participating in this study it is possible that you may experience some discomfort. The vaccination may cause mild/moderate fever, injection-site pain, redness, soreness, itchiness,or other discomfort. Allergic reactions may be observed occasionally. Generally, these reactions can disappear within 24 hours without treatment. There is a very small chance of infection.

The blood taking may also be accompanied by some risk. Pain, bleeding, bruising, or feeling lightheaded or mild pain may occurred. Syncope or infection at the blood drawing site israre.

Unintended release of some test results such as HLA type could be used by insurers or employers, which may suggest risk of disease for you or your family. Therefore the genetic test results are kept confidential, we are not going to release the results to you or any unauthorized person.

While the possibility of serious adverse reaction happening due to participating is very low, you should still be aware of the possibility. If something unexpected happens and harm does occur, we will provide you appropriate treatment and corresponding economic compensation.

Previous studies have indicated that immunization with Ad5 vector based or other viral vector based vaccines may increase the risk of HIV-1 acquisition, so we suggest you avoid HIV associated high-risk behavior such as dangerous sexual behavior, intravenous drug use, or illegal blood transfusion.

9. Possible benefits for taking part in this study

All the laboratory examination and vaccinations in study are free. By taking part in this trial, you may develop the immune response to the Ebola virus due to the vaccinations. But the vaccination cannot provide 100% guarantee for resistance against Ebola virus disease. You may not get other healthy benefit for taking part in this study, but your contribution may benefit other people and the whole nation.

10. Reimbursements

In order to compensate your travel expenses, blood denotation and the time lost for participating in this study, you will get a reimbursements of 500 RMB after each visit, and after complete all the 8 visits you will get a total of 4000 RMB as reimbursements. Relevant medical expenses for youto take part in this study are free, such as expense of vaccine, laboratory examination, and so on. You will not get other payments unrelated to the trial.

 

11. Confidentiality of the study data

The results of this study will be given to the sponsor. Your personal information will be kept confidentially. Your name will not appear in any published information on the study or report. In addition, the representative of the China State Food and Drug Administration (SFDA) may examine the medical history record related to this study, so as to verify the accuracy of the data collected in this study, but it will not involve your detailed personal information.

 

12. Sharing of the results

Confidential information will not be shared. The test result in this study that we get will not be shared with you, unless the results indicate there is a potential safety risk and you need to know. But if you want, you can require the doctors to give you the results.We will publish the results in order that other interested people may learn from our research. The identity of subjects remains confidential to protect your privacy.

 

13. Right to Refuse or Withdraw

You may choose not to take part. You can change your mind and withdraw (drop out) later. There will be no penalty, and you will not lose any benefits they receive now or have a right to receive proper treatment.

We will tell you if we learn new information that could change your mind about taking part or continuing in this research study. If you want to drop out, you should tell us. We will make sure you can end the study in the safest way.

The study doctor or the study sponsor may decide to let you out of the study without your agreement if, for example:

You do not follow the directions of the study team, or;

The study doctor decides that the study is not in your best interest, or;

The study is stopped by the study sponsor, the institutional review board (IRB), or by a regulatory agency.

The decision of the participation in the study is entirely voluntary. You may withdraw from the study without giving any reason at any time before the study or after it starts. If you withdraw the study for any reason, biological samples (for example, blood samples) that have been collected from you (but not yet fully analyzed) can be destroyed by making a request to the study doctor. However, any data already generated from your samples will be kept to preserve the value of the study. If you leave the study for any reason, you will not need to donate blood or receive any further tests.

 

14. Alternatives to participating

Because there is no Ebola vaccines available in the market at present. If you do not wish you to take part in this study, no other intervention can provide to you.Since there is no Ebola infected patient has been found in China, you may not have the risk of exposure if you stay in China. You can also take some nonspecific precautionary measures such as hand-washing to protect yourself.

 

15. Contact

If you have any questions you may ask the investigator or the study staff now or later, even after the study has started. If you wish to ask questions later, you may contact any of the following:

Feng-Cai ZhuTel: +86-25-83759418

 

This proposal has been reviewed and approved by the IRB of Jiangsu Provincial Center for Disease Prevention and Control, which is a committee whose task is to make sure that research participants are protected from harm. If you wish to find about more about the IRB, contact:

Hui-Yuan Cai Tel: +86-25-83759406

 

16. Certificate of Consent

By signing this Informed Consent Form, you confirm that you have read this information and have sufficiently considered it, that the doctor has explained this study to you, that your questions have been answered, that you have had enough time to make the decision on the participation, and that you agree to take part in this study. You have learned that you may withdraw from this study at any time without any reason, and the withdrawal from the study will not affect the present or future treatment of you.

 

Name of participant: ____( completed by doctor)

Signature of participant:__ ______       Date:_____ _____ _____( completed by participant)

                                        year/month/day

 

17. Statement of investigator

I confirm that I have explained the content, procedures, possible risks and benefits of this study to the participant, and have given him or her adequate answers about any question, and the family members of the participant have got satisfactory answers.

 

Signature of doctor:_____                                 Date:____ ____ ____

year/month/day


 

15.    SIGNATURES

SPONSOR: Beijing Institute of Biotechnology, Tianjin CanSino Biotechnology Inc.

This Study Protocol was subjected to critical review and has been approved.

Clinical Development Manager:  [Signature]

Wei Chen

Beijing Institute of Biotechnology.

Date

LABORATORY TEST: 

Responsible Officer:           [Signature]

Gui-Rong Zhang

Beijing Institute for Drug and Instrument Quality Control

Date

 

PRINCIPLE INVESTIGATOR:

I have reviewed this Study Protocol, including Appendices. I will conduct the clinical study asdescribed and will adhere to GCP and all the ethical and regulatory requirements stated.

I have read and understood the contents of the Investigator's Brochure.

 

Principal Investigator:    [Signature]                 

Professor Feng-Cai Zhu

Jiangsu Provincial Center for Disease Control and Prevention

Date


 

16.    REFERENCE

1.      Feldmann H, Klenk HD. Marburg and Ebola viruses. Adv Virus Res 1996; 47: 1-52.

2.      Balter M. Emerging diseases. On the trail of Ebola and Marburg viruses. Science 2000; 290(5493): 923-5.

3.      Baize S, Pannetier D, Oestereich L, et al. Emergence of Zaire Ebola virus disease in Guinea. N Engl J Med 2014; 371(15): 1418-25.

4.      Kucharski AJ, Edmunds WJ. Case fatality rate for Ebola virus disease in west Africa. Lancet 2014; 384(9950): 1260.

5.      Smither SJ, Eastaugh LS, Steward JA, Nelson M, Lenk RP, Lever MS. Post-exposure efficacy of oral T-705 (Favipiravir) against inhalational Ebola virus infection in a mouse model. Antiviral Res 2014; 104: 153-5.

6.      Organization WH. What this – the largest Ebola outbreak in history – tells the world. 2014. http://www.who.int/csr/disease/ebola/ebola-6-months/lessons/en/ (accessed December 23 2014).

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