Observer Blinded, Randomized Trial to Evaluate Safety and Immunogenicity of a Novel Vaccine Formulation MV-ZIKA-RSP
Phase: PHASE1
Conditions: Zika Virus Infection
Sponsor: Themis Bioscience GmbH

Official Protocol Title: 
NCT number: 
NCT04033068 
Document Date: 
28-Sep-2020
Observer blinded, randomized trial to evaluate safety and
immunogenicity of a novel vaccine formulation MV-ZIKA-RSP

3DJH
3URWRFRO09=,.$563
(XGUD&7
2EVHUYHUEOLQGHGUDQGRPL]HGWULDOWRHYDOXDWHVDIHW\DQG
LPPXQRJHQLFLW\RIDQRYHOYDFFLQH IRUPXODWLRQ 09=,.$563
6SRQVRU
7KHPLV%LRVFLHQFH*PE+
0XWKJDVVH
9LHQQD$XVWULD
7HO
3URWRFRO9HUVLRQ

9HUVLRQ'DWH
 6HSWHPEHU
&OLQLFDO3KDVH
3KDVH,
&52
1HR[&OLQLFDO5HVHDUFK
&&93
PPD
*)

&&93
PPD
PPD
PPD
PPD
*)

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH
,QYHVWLJDWRU¶V$SSURYDO

,FRQILUPWKDW,KDYHUHDGWKHSURWRFRODQGWKDW,DJUHHWRFRQGXFWWKLVVWXG\LQDFFRUGDQFHZLWK
WKHSURWRFRO,QWHUQDWLRQDO&RQIHUHQFHRQ +DUPRQLVDWLRQDQG*&3JXLGHOLQHVDQGZLWKWKH
DSSOLFDEOHORFDOUHJXODWRU\UHTXLUHPHQWV0RUHRYHUWKHVLWHZLOONHHSDOOLQIRUPDWLRQREWDLQHG
IURPWKHSDUWLFLSDWLRQLQWKLVVWXG\FRQILGHQWLDOXQOHVVRWKHUZLVHDJUHHGLQZULWLQJ


,19(67,*$725











'DWH

6LJQDWXUH













,QVWLWXWH RI 6SHFLILF 3URSK\OD[LV DQG 7URSLFDO
0HGLFLQH0HGLFDO8QLYHUVLW\9LHQQD
.LQGHUVSLWDOJDVVH9LHQQD$XVWULD
3KRQH





8QLY3URILQ'ULQ8UVXOD:LHGHUPDQQ6FKPLGW




$GGUHVV

1DPH


&&93
PPD
*)

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 4/82 
1. 
STUDY SYNOPSIS 
Protocol Number 
MV-ZIKA-RSP-101 
Title of the Study 
Observer blinded, randomized trial to evaluate safety and 
immunogenicity of a novel vaccine formulation MV-ZIKA-RSP  
Study Centres 
One site in Vienna, Austria 
Study Period 
Study start (FSI) August 2019  
Study Phase 
Clinical Phase I 
Objectives 
Primary Objective 
- To investigate safety and tolerability of MV-ZIKA-RSP, a 
novel vaccine formulation, up to day 56 after first 
immunization consisting in one or two vaccinations 
Secondary Objective 
- To investigate long-term safety, up to study day 182, after 
first immunization  
- 
To investigate immunogenicity up to day 56 after first 
immunization consisting in one or two vaccinations with 
different doses 
- To get information about the optimal dose of MV-ZIKA-RSP 
vaccine regarding safety, tolerability, and immunogenicity 
- To investigate specific cell mediated immunity induced by all 
different treatment groups 
Study Design 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Observer blinded, block-randomized, dose finding, single centre, 
phase I trial in 48 participants. 
After completion of screening procedures, participants are 
randomized to one of four treatment groups (A, B, C or D): 
- A: 14 participants will receive two high dose treatments with 
MV-ZIKA-RSP- 1 x105 (0.5 log) TCID50 /dose on day 0 and 
28  
- B: 14 participants will receive two low dose treatment with 
MV-ZIKA-RSP- 2,5 x104 (0.5 log) TCID50 on day 0 and 28  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 5/82 
- C: 12 participants will receive one high dose treatment with 
MV-ZIKA-RSP- 1 x105 (0.5 log) TCID50 /dose on day 0 and 
placebo on day 28 
- D: 8 participants will receive placebo on day 0 and day 28 
After the screening visit (V0), participants will perform three 
additional visits to the site. On study days 0 and 28 participants will 
receive the vaccination, as well as, safety assessments together with 
immunogenicity sample collection. On study day 56 no vaccination 
will be done, but safety assessments and sample collection are 
scheduled. Safety follow-up assessments will be done by a phone 
call (Visit 4) approximately 6 months (day 182) after the first 
vaccination. A preliminary analysis will be performed after all study 
subjects completed Visit 3 (day 56).  
Number of 
Participants 
48 healthy participants 
Main Inclusion Criteria Healthy female or male volunteers, 18 - 55 years old at screening. 
Investigational 
Medicinal Product 
MV-ZIKA RSP, presented as a liquid frozen (lf) formulation, is a live 
recombinant measles virus vaccine expressing ZIKA virus antigens; 
suspension for injection; administered by i.m. injection, used in two 
different dose levels: 
- High dose: 1 x105 (+/- 0.5 log) TCID50  
- Low dose: 2,5 x104 (+/- 0.5 log) TCID50  
Placebo 
Physiological saline solution (0.9% NaCl), administered by i.m. 
injection 
Duration of the Study 
Study duration per subject will be 182 days (6 months), consisting of 
4 weeks treatment period (study day 0 till study day 28) and 22 
weeks follow up period (study day 28 till study day 182). The overall 
study duration is estimated to be approximately 18 months, from 
study approval until reporting. 
Statistical Methods 
The primary analysis will compare adverse events rates up to day 56 
between treatment groups using Fisher exact tests.  
Solicited AEs, unsolicited AEs and SAEs will be summarized by 
severity and term up to day 56 in the preliminary analysis and AE up 
to 182 are summarized in the final analysis. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 6/82 
The immunogenicity analysis will compare the anti-zika-rsp VNT 
antibody geometric mean titre (GMT) between the treatment groups. 
GMTs and GMT ratios will be estimated by applying an analysis of 
variance including the factor treatment group. This will be done using 
log10 transformed data and taking the anti-log of the resulting point 
estimates for the least squares means, least squares means 
differences and the corresponding 2-sided 95% CIs.  
P-values will also be provided to compare GMTs between the 
treatment groups adjusted for multiple comparisons according to 
Tukey-Kramer. Seroconversion rates will be compared between 
groups using Fisher’s exact test. 
 
Criteria for Evaluation 
Primary endpoint  
- Rate of adverse events (AEs) up to study day 56 
Secondary endpoints: 
- Rate of solicited and unsolicited AEs, as well as, reported 
serious adverse events (SAEs) up to study day 182 (long-
term safety) compared between the 4 different treatment 
groups.  
- Immunogenicity on study days 0, 28 and 56 as confirmed by 
the presence of functional anti-zika antibodies as determined 
by VNT and by ELISA 
- Cell-mediated immunity specific for ZIKA-RSP, up to day 56, 
as confirmed by the presence of specific functional CD4+ and 
CD8+ T-cells 
- Safety 
laboratory 
parameters 
(haematology, 
serum 
chemistry, urinalysis) 
Benefit/Risk 
Assessment 
 
The measles virus vector is well known. Possible risks that are 
frequently associated with vaccinations are the occurrence of local 
reactions e.g. oedema, induration and erythema, transient local 
pain or tenderness at the injection site as well as mild to moderate 
headache, myalgia, flu-like symptoms or fatigue. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 7/82 
Like any other vaccine, MV-ZIKA-RSP might induce allergic and 
anaphylactic reactions, apart from the described local reactions at 
the vaccination site and systemic flu-like reactions. In rare cases the 
injection can lead to a vasovagal reaction immediately after injection 
of the vaccine. The needle pricks for blood sampling may also cause 
local reactions such as oedema. The participants cannot expect 
direct benefit from study participation, except for potential boosting 
of measles immunity and increasing immunity to zika. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 8/82 
2. 
LIST OF ABBREVIATIONS 
Ab 
Antibody 
AE 
Adverse Event 
AESI 
Adverse Event of special interest 
AGES 
Österreichische Agentur für Gesundheit und Ernährungssicherheit GmbH 
AMG 
"Arzneimittelgesetz" [Austrian Law on Regulation of Therapeutic Goods] 
ALT 
Alanine Aminotransferase (SGPT) 
aPTT 
Abbreviated Partial Thrombin Time 
AST 
Aspartate Aminotransferase (SGOT) 
CA 
Competent Authority 
CCID50 
Cell Culture Infectious Dose 50% 
CHIKV 
Chikungunya Virus 
CRO 
Clinical Research Organisation 
CV 
Curriculum Vitae 
DSMB 
Data Safety Monitoring Board 
(e)CRF 
(electronic) Case Report Form 
EC 
Ethics Committee 
ELISA 
Enzyme Linked Immunosorbent Assay 
ER 
Emergency Room 
FDA 
(United States) Food and Drug Administration 
FSI 
First Subject In 
GCP 
Good Clinical Practice 
GMO 
Genetically Modified Organism 
GMT 
Geometric Mean Titre 
HSA 
Human Serum Albumin 
HBV 
Hepatitis B Virus 
HCV 
Hepatitis C Virus 
HIV 
Human Immunodeficiency Virus 
HSD 
Honest Significant Difference 
IEC 
Independent Ethics Committee 
i.m. 
Intra Muscular 
IB 
Investigator’s Brochure 
ICH 
International Conference on Harmonisation 
IMP 
Investigational Medicinal Product 
IRB 
Institutional Review Board 
ITT 
Intent-To-Treat 
MAE 
Medically-attended Adverse Event 
MedDRA 
Medical Dictionary for Regulatory Activities 
mITT 
Modified Intent-To-Treat 
Mo 
Month 
MV 
Measles Virus 
NHP 
Non-Human Primate 
PBMC 
Peripheral Blood Mononuclear Cell 
PP 
Per-Protocol 
PRNT50 
Plaque Reduction Neutralisation Test 50% 
PT 
Prothrombin Time 
SAE 
Serious Adverse Event 
SAR 
Serious Adverse Reaction 
SPS 
Stabilising and Protecting Solutions 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TCID50 
Tissue Culture Infective Dose 50% 
VNT 
Virus Neutralization Test 
WHO 
World Health Organisation 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 9/82 
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 10/82 
3. 
TABLE OF CONTENTS 
1. 
STUDY SYNOPSIS ................................................................................................... 4 
2. 
LIST OF ABBREVIATIONS ........................................................................................ 8 
4. 
BACKGROUND INFORMATION AND RATIONAL ...................................................15 
4.1 
Investigational Drug ............................................................................................. 15 
4.2 
Disease Background ............................................................................................ 15 
4.3 
Preclinical Studies ................................................................................................ 17 
4.3.1 
Non-Clinical Pharmacology Studies Performed .............................................17 
4.3.2 
Primary Pharmacodynamics ..........................................................................17 
4.4 
Clinical Studies .................................................................................................... 20 
4.4.1 
Phase I Study ................................................................................................20 
4.5 
Study Rationale .................................................................................................... 25 
4.6 
Benefit/Risk Assessment ..................................................................................... 25 
5. 
TRIAL OBJECTIVES AND MEASUREMENTS .........................................................28 
5.1 
Primary Objective ................................................................................................. 28 
5.2 
Secondary Objectives .......................................................................................... 28 
6. 
TRIAL DESIGN .........................................................................................................29 
6.1 
Overall Study Design ........................................................................................... 29 
6.2 
Study Endpoints ................................................................................................... 29 
6.2.1 
Primary Endpoint ...........................................................................................29 
6.2.2 
Secondary Endpoints ....................................................................................29 
6.3 
Treatment Groups ................................................................................................ 30 
6.4 
Study Flowchart ................................................................................................... 31 
6.5 
Study Procedures ................................................................................................ 31 
7. 
PARTICIPANT IDENTIFICATION .............................................................................33 
7.1 
Trial Participants .................................................................................................. 33 
7.2 
Inclusion Criteria .................................................................................................. 33 
7.3 
Exclusion Criteria ................................................................................................. 33 
8. 
TRIAL PROCEDURES .............................................................................................36 
8.1 
Informed Consent (AMG § 38) ............................................................................. 36 
8.2 
Randomisation ..................................................................................................... 36 
8.3 
Blinding and Code-breaking ................................................................................. 37 
8.4 
Screening Procedures .......................................................................................... 37 
8.4.1 
Screening Visit (VS, 1-14 days prior to Visit 1) ..............................................38 
8.5 
Treatment Period ................................................................................................. 38 
8.5.1 
Visit 1 (day 0) ................................................................................................38 
8.5.2 
Post Dose Safety Follow-up (day 7 1 days) .................................................39 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 11/82 
8.5.3 
Visit 2 (day 28 3 days) .................................................................................39 
8.5.4 
Visit 3 (day 56 3 days) .................................................................................40 
8.6 
Post Treatment Safety Follow up Period .............................................................. 40 
8.6.1 
Visit 4 (day 182 14 days) .............................................................................40 
8.7 
Early Termination Visit (ET) ................................................................................. 40 
8.8 
Unscheduled Visits .............................................................................................. 41 
8.9 
Discontinuation/Withdrawal of Participants ........................................................... 41 
8.9.1 
Safety Stopping Rules ...................................................................................41 
8.9.2 
Replacement of Participants ..........................................................................43 
8.10 
End of Study ........................................................................................................ 43 
8.11 
Handling of Samples ............................................................................................ 43 
8.11.1 
Samples for Safety Laboratory Parameters ...................................................43 
8.11.2 
Immunogenicity Samples ..............................................................................44 
8.11.3 
Pregnancy Test – Contraception ...................................................................45 
8.11.4 
Labelling, Storage and Transport of Samples ................................................47 
9. 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP) ..................................................48 
9.1 
Investigational Medicinal Product Description ...................................................... 48 
9.1.1 
MV-ZIKA-RSP vaccine-test product ..............................................................48 
9.1.2 
Placebo .........................................................................................................48 
9.2 
Packaging and Labelling ...................................................................................... 49 
9.3 
Dose Regimen ..................................................................................................... 49 
9.4 
Justification of Clinical Dose Regime ................................................................... 49 
9.5 
Storage and Transport Conditions........................................................................ 50 
9.6 
IMP Preparation and Distribution ......................................................................... 51 
9.7 
GMO Handling ..................................................................................................... 51 
9.8 
Route and Method of Administration of IMP ......................................................... 51 
9.9 
Treatment Compliance ......................................................................................... 51 
9.10 
Study Drug Accountability .................................................................................... 52 
9.11 
Prior and Concomitant Therapy ........................................................................... 52 
9.11.1 
Permitted Prior and Concomitant Therapy .....................................................52 
9.11.2 
Forbidden Prior and Concomitant Therapy ....................................................52 
10. 
SAFETY REPORTING ..............................................................................................54 
10.1 
Definitions ............................................................................................................ 54 
10.1.1 
Adverse Event (AE) .......................................................................................54 
10.1.2 
Clinical Laboratory Adverse Event .................................................................54 
10.1.3 
Adverse Reaction (AR) ..................................................................................54 
10.1.4 
Serious Adverse Event (SAE)........................................................................54 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 12/82 
10.1.5 
Suspected Unexpected Serious Adverse Reaction (SUSAR) ........................55 
10.1.6 
Causality .......................................................................................................55 
10.1.7 
Severity .........................................................................................................55 
10.1.8 
Expectedness ................................................................................................56 
10.1.9 
Outcome .......................................................................................................56 
10.2 
Recording of Adverse Events ............................................................................... 57 
10.3 
Reporting of Serious Adverse Events ................................................................... 58 
10.4 
Follow-up of Adverse Events ................................................................................ 58 
10.5 
Reporting of SUSARs .......................................................................................... 59 
10.6 
Reporting of Pregnancies ..................................................................................... 59 
10.7 
Development Safety Update Reports ................................................................... 60 
11. 
ASSESSMENT OF SAFETY .....................................................................................61 
11.1 
Safety Laboratory Parameter Assessments ......................................................... 61 
11.2 
Symptom-directed Physical Examination ............................................................. 61 
11.3 
System-based Physical Examination ................................................................... 61 
11.4 
Vital Signs and Body Temperature ....................................................................... 61 
11.5 
Post Vaccination Reactogenicity Assessments .................................................... 61 
11.6 
Local and Systemic Tolerability ............................................................................ 62 
11.7 
Subject Diary ....................................................................................................... 62 
12. 
STATISTICS .............................................................................................................64 
12.1 
Preliminary Analysis ............................................................................................. 64 
12.2 
Methods of Statistical Immunogenicity Analysis ................................................... 64 
12.3 
Methods of Statistical Safety Analysis .................................................................. 64 
12.4 
Determination and Justification of Sample Size.................................................... 65 
12.5 
Analysis Populations ............................................................................................ 65 
12.5.1 
Safety Population ..........................................................................................65 
12.5.2 
Modified Intent-to-Treat (mITT) Population ....................................................65 
12.5.3 
Per-Protocol (PP) Population ........................................................................65 
12.6 
Statistical Analysis Plan ....................................................................................... 66 
12.7 
Missing Data ........................................................................................................ 66 
13. 
QUALITY CONTROL AND QUALITY ASSURANCE .................................................67 
13.1 
Periodic Monitoring .............................................................................................. 67 
13.2 
Audit and Inspection ............................................................................................ 67 
14. 
ETHICS ....................................................................................................................68 
14.1 
Ethical Conduct of the Study ................................................................................ 68 
14.2 
Ethics Committees (EC) (AMG §§ 40,41) ............................................................. 68 
14.3 
Competent Authority (CA) .................................................................................... 68 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 13/82 
15. 
DATA HANDLING AND RECORD KEEPING ...........................................................69 
15.1 
Source Data and Records .................................................................................... 69 
15.2 
Periodic Monitoring .............................................................................................. 69 
15.3 
Access to Data ..................................................................................................... 69 
15.4 
Data Collection..................................................................................................... 70 
15.5 
Electronic Case Report Forms (eCRFs) ............................................................... 70 
15.4.1 
eCRF Entries .................................................................................................70 
15.4.2 
Changes to eCRF Data .................................................................................70 
15.4.3 
eCRF Entry Validation ...................................................................................71 
15.5 
Confidentiality of Subject’s Data ........................................................................... 71 
15.6 
Record Maintenance, Archiving and Destruction .................................................. 71 
16. 
CHANGES IN THE CONDUCT OF THE STUDY ......................................................73 
16.1 
Protocol Amendments .......................................................................................... 73 
16.2 
Study Termination ................................................................................................ 73 
17. 
DEVIATIONS FROM THE PROTOCOL ....................................................................74 
17.1 
Relevant Protocol Deviations ............................................................................... 74 
17.2 
Serious Breaches ................................................................................................. 74 
17.3 
Premature Subject Withdrawal ............................................................................. 74 
18. 
FINANCE AND INSURANCE ....................................................................................75 
18.1 
Contractual and Financial Details ......................................................................... 75 
18.2 
Insurance (AMG §32(2)) ...................................................................................... 75 
19. 
REPORTING AND PUBLICATION ...........................................................................76 
19.1 
Clinical Study Report............................................................................................ 76 
19.1 
Publication Policy ................................................................................................. 76 
20. 
REVISION INDEX .....................................................................................................77 
21. 
REFERENCES .........................................................................................................78 
22. 
APPENDICES ...........................................................................................................82 
 
 
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 14/82 
List of Tables: 
 
Table 1. Treatment groups and vaccination schedule ...........................................................30 
Table 2. Study procedures ...................................................................................................31 
Table 3. Blood sample collection ..........................................................................................45 
Table 4. Dose regime ...........................................................................................................50 
Table 5. Grading of local reactions .......................................................................................62 
Table 6. Grading of Systemic Reactions ...............................................................................63 
 
 
 
 
 
06CCVP
06G6F2

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH

%$&.*5281',1)250$7,21$1'5$7,21$/

,QYHVWLJDWLRQDO'UXJ
7KH09=,.$563YDFFLQHFDQGLGDWHLVDOLYHDWWHQXDWHGUHFRPELQDQWYLUDOYHFWRUHGYDFFLQH
IRUSURSK\OD[LVRI=LNDYLUXVLQIHFWLRQ

FKHPLFDOO\
V\QWKHVL]HGDQGLQVHUWHGLQWRWKH6FKZDU]YDFFLQHVWUDLQRIPHDVOHVYLUXV09WRSURGXFHWKH
FDQGLGDWHYDFFLQH09=,.$563
DQGFROOHDJXHVDWWKH3DVWHXU,QVWLWXWHLQ3DULVUHFHQWO\LQWURGXFHGWKHOLYH
DWWHQXDWHG09YDFFLQH6FKZDU]VWUDLQ±096FKZDVDYHFWRUWRH[SUHVVKHWHURORJRXVYLUDO
DQWLJHQV>GHVFULEHGLQGHWDLOLQ&RPEUHGHW@7KLVWHFKQRORJ\FDQEHXVHGDVDSODWIRUP
IRU WKH JHQHUDWLRQ RI SURSK\ODFWLF YDFFLQHV DJDLQVW GLIIHUHQW LQGLFDWLRQV LH HPHUJLQJ
LQIHFWLRXVGLVHDVHV
7KH09YHFWRUKDVEHHQVKRZQWRVWDEO\H[SUHVVODUJHKHWHURORJRXVDQWLJHQVXSWRNEORQJ
7RGDWHWKHYHFWRUKDVEHHQXVHGWRJHQHUDWHVHYHUDOUHFRPELQDQW09FORQHVH[SUHVVLQJ
KHWHURORJRXVYLUDODQWLJHQVLQFOXGLQJ&KLNXQJXQ\D9LUXV&+,.9%UDQGOHU:HVW1LOH
9LUXV:19>'HVSUqV%UDQGOHU@'HQJXHYLUXV'(19>%UDQGOHU%UDQGOHU
@DQGKXPDQLPPXQRGHILFLHQF\YLUXV+,9>/RULQ@
7KH PHDVOHV YHFWRUEDVHG &KLNXQJXQ\D YDFFLQH LV DGYDQFHG WR ODWH VWDJH FOLQLFDO
GHYHORSPHQW 7KH &KLNXQJXQ\D YDFFLQH ZDV WKH ILUVWPHDVOHVYHFWRUHG LQIHFWLRXV GLVHDVH
FDQGLGDWHWHVWHGLQKHDOWK\DGXOWVXEMHFWV5DPVDXHUHWDO$3KDVHFOLQLFDOWULDOZDV
UHFHQWO\FRPSOHWHGVKRZLQJWKHVDIHW\DQGLPPXQRJHQLFLW\RIWKHYDFFLQHFDQGLGDWHLQ
KHDOWK\DGXOWVXEMHFWV5HLVLQJHUHWDO
$=LNDYDFFLQHFDQGLGDWHZDVSUHYLRXVO\GHYHORSHGE\WKHVWXG\VSRQVRU7KHPLV$3KDVH
 FOLQLFDO WULDO ZDV FRQGXFWHG LQ $XVWULD LQ   1&7
GHWDLOVDUHGHVFULEHGEHORZ
)RUDGHWDLOHGGHVFULSWLRQRIWKHYDFFLQHFRQVWUXFWDQGPHFKDQLVPRIDFWLRQSOHDVHUHIHUWRWKH
FXUUHQWYHUVLRQRIWKH09=,.$563LQYHVWLJDWRUV¶EURFKXUH

'LVHDVH%DFNJURXQG
=LNDYLUXVLVDQHPHUJLQJPRVTXLWRERUQHIODYLYLUXVLQLWLDOO\LVRODWHGIURPDUKHVXVPRQNH\LQ
WKH=LNDIRUHVWLQ8JDQGDLQ*XEOHU'LFN7KHILUVWKXPDQLQIHFWLRQZDV
UHSRUWHGLQ1LJHULDLQ0DFQDPDUD/LNHGHQJXHDQGFKLNXQJXQ\DYLUXVHV=LND
YLUXVDGDSWHGIURPDQDQFHVWUDOWUDQVPLVVLRQF\FOHLQYROYLQJQRQKXPDQSULPDWHVDQGDEURDG
&&93
PPD
CCI
*)

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 16/82 
spectrum of forest mosquito species as vectors to an urban cycle involving humans as 
reservoirs and the widely distributed Aedes mosquitoes as vectors. Since the 1950s, Zika virus 
had only been reported as circulating sporadically in Africa and Southeast Asia. In 2007, Zika 
virus was isolated for the first time in the Pacific, on the Micronesian island of Yap (Duffy, 
2009). Between October 2013 and April 2014, French Polynesia experienced the largest Zika 
outbreak ever reported at that time (Cao-Lormeau, 2013). More than 32000 patients were 
suspected for Zika virus infection. Between 2014 and 2015, Zika virus had spread to other 
Pacific islands, notably the Cook Islands and Easter Island (Chile). In March 2015, Brazil 
reported the autochthonous transmission of Zika virus (Zanluca, 2015) and declared an 
unprecedented outbreak 6 months later with preliminary estimates of 440,000 to 1.3 million 
cases of infection through December 2015 (European Centre for Disease Prevention and 
Control, 2015). Since the outbreak in Brazil the virus has been found in over 60 countries and 
territories worldwide (Center for Disease Control and Prevention, Nov 2016). Although WHO 
has declared an end to its global health emergency in Nov 2016, the Zika crisis is not 
considered over. The virus is still spreading, and it remains a global health threat. 
The current Zika epidemic is the largest epidemic ever recorded for this virus. Usually, a Zika 
infection is mild with symptoms such as fever, rash, conjunctivitis and joint pain lasting for 
several days to a week. People often do not realize they have been infected. Symptoms of 
Zika are often like other viruses spread through mosquito bites, like dengue and chikungunya. 
Although infection with Zika virus was usually associated to mild disease, its emergence in the 
Americas has coincided with a steep increase in patients developing Guillain-Barré syndrome. 
Moreover, infection with Zika virus has been linked to the birth of babies with neurological 
complications, in particular congenital microcephaly (WHO, 2016; ECDC 2015; Soares de 
Araújo, 2016), and it was shown that when pregnant women are exposed to Zika virus during 
the first trimester of pregnancy, the risk of microcephaly for the newborn is increased 50 times 
from 2/10 000 to 1/100 (Cauchemez, 2016). In Feb 2016, the WHO declared the suspected 
link between Zika virus and neurological disorders and neonatal malformations a Public Health 
Emergency of International Concern. 
In this context, in March 2016, experts gathered at WHO agreed that the development of a 
preventive vaccine is a major priority to respond to Zika epidemics in the future (WHO, Media 
centre, 2016). A clear target product profile (TPP) for this vaccine is still under work, but 
pragmatic strategies were asked to fast track the development of a safe an effective product. 
Due to the established link between Zika virus infection and the appearance of congenital 
microcephaly in babies born to infected mothers, one could argue that a Zika vaccine has to 
be suitable for use in pregnant women. However, no licensed vaccine is currently 
recommended for the use in pregnant women. Moreover, with the demonstrated association 
of Zika infection with Guillain-Barré syndrome, the observation of possible sexual transmission, 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 17/82 
and the appearance of developmental defects probably appearing very early in pregnancy, it 
is very likely that the TPP will be addressed to the general population. In any case a Zika 
vaccine will have to demonstrate an excellent safety profile, particularly concerning the risk of 
neurotropism. 
4.3 
Preclinical Studies  
Brief Summaries of preclinical studies are given below. These studies are described in more 
detail in the current version of the MV-ZIKA-RSP investigators’ brochure. 
4.3.1 Non-Clinical Pharmacology Studies Performed 
The immunogenicity and efficacy of MV-ZIKA-RSP vaccine candidates were examined in non-
human primates and in transgenic CD46/IFNAR mice (CD46tg; IFNAR -/-), an animal model 
for the MV vector system. The vaccine immunogenicity was determined in mice by Enzyme-
linked immuno-sorbent assay (ELISA) and plaque reduction neutralization assays (PRNT) 
from the sera of immunized mice. In addition, we showed the vaccine efficacy in two mouse 
challenge models, one lethal model and one disease model. In both models, all mice were 
protected against ZIKAV challenge. 
Furthermore, the vaccine immunogenicity and efficacy were confirmed in non-human primates, 
which is a more relevant, immunocompetent animal model. 
4.3.2 Primary Pharmacodynamics  
Analysis of the immunogenicity and protective efficacy of MV-ZIKA_RSP vaccine candidate in 
CD46-IFNAR mice after prime/boost immunization and subsequent Zika virus challenge model 
CD46-IFNAR mice were immunized twice intraperitoneally (ip) with MV-ZIKA RSP (106 TCID50) 
one month apart. Control mice were immunized with the same dose of empty Schwarz vaccine 
strain of MV vector. Subsequently, one month after final immunization the mice were 
challenged by ip injection of 105 TCID50 of an African Zika virus strain (Zika AFR). Blood 
samples were collected 1 month after each immunization. The sera were analyzed for the 
presence of functional, neutralizing antibodies by plaque reduction neutralization test (PRNT). 
In addition, enzyme-linked immunosorbent assays (ELISAs) were performed to detect anti-
ZIKV binding antibodies. All animals that received two doses of the MV-ZIKA_RSP vaccine 
were protected against lethal challenge and did not show any signs of disease. 
An additional animal model was used to show Zika virus viremia in the measles mouse model. 
Animals were immunized with two doses of MV-ZIKA RSP (106 TCID50) at day 0 and day 28, 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 18/82 
followed by challenge with Zika virus (South African Strain). This strain causes viremia but no 
signs of disease in the mouse model. The Zika virus viremia after challenge was analyzed at 
different time points (day 1, 2, 3, 5, 7, 10, and 21) by determination of viral ZIKV RNA in blood 
samples by quantitative PCR. Again, all animals that received the MV-ZIKA_RSP vaccine 
showed very high titers of neutralizing antibodies. Accordingly, were protected against Zika 
virus viremia.   
Protective efficacy of the MV-ZIKA_RSP vaccine in non-human primates. 
The vaccine efficacy and safety were demonstrated in a mouse model that allows measles 
virus replication due to a deficiency in the Interferon system. Thus, the vaccine immunogenicity 
and efficacy were tested in non-human primates (Cynologous Macaques). This animal model 
is frequently used for vaccine studies with measles vectored vaccines as well as for Zika 
disease models. 
Eight animals per groups were immunized with MV-ZIKA_RSP or with the measles vector 
backbone MV-Schwarz. The animals received two MV-ZIKA RSP immunizations at 105 TCID50, 
4 weeks apart. The immunogenicity was demonstrated at 4 weeks after the first and the second 
dose. All animals that received MV-ZIKA_RSP showed seroconversion to anti-ZIKA antibodies 
by ELISA and all animals showed high levels of neutralizing antibodies.  
Seventeen weeks after the first dose all animals were challenged with 1x104 TCID50 Zika virus. 
All animals in the MV-Schwarz groups showed a viremia between days 1 -9 after challenge. 
The viremia in the serum was as high as 1x107 RNA copies per mL serum. In contrast, animals 
that received the MV-ZIKA_RSP vaccine were fully protected against challenge. Three out of 
eight animals showed a small increase in virus copy numbers at peak viremia on day 4, but 
the levels were at or below the limit of quantification.  
In summary, the MV-ZIKA vaccine induced a strong and protective immune response in a 
relevant animal’s model.  
Repeated-dose toxicity study by intramuscular route in cynomolgus monkeys followed by a 3-
week treatment free period 
The objective of this study was to evaluate the potential toxicity of the test item, MV-ZIKA RSP, 
following an immunization schedule corresponding to three intramuscular administrations of 
the Intended Human Dose (IHD), administered every 2 weeks, to cynomolgus monkeys. On 
completion of the treatment period, designated animals were held for a 3-week treatment-free 
period in order to evaluate the reversibility of any findings or potential delayed effects. 
One group of six male and six female cynomolgus monkeys received the test item MV-ZIKA 
RSP, by intramuscular administration on Days 1, 15 and 29 at the maximal intended human 
dose of 2 x 106 TCID50/injection. A control group of three animals per sex received the control 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 19/82 
item (sterile saline), under the same experimental conditions. Two days after the last injection 
(i.e. Day 31), the animals were euthanized (early sacrifice), except for three animals per sex 
in the test item-treated group which were kept for a 3-week treatment-free period and 
euthanized on Day 50 (late sacrifice). 
Measles and Zika serological status of each animal were checked before allocation to the study 
and only seronegative animals were included. The animals were checked daily for mortality 
and clinical signs. Local tolerance at the injection sites was assessed on each day of 
administration, before, 4 and 24 hours after administration. Rectal temperature was recorded 
on Days 1 and 29, before dosing, 4 and 24 hours after administration. Body weight was 
recorded pre-test, on the first day of administration and then daily during the week after each 
administration, and at least twice weekly until the end of the study. Group food consumption 
was checked daily. Ophthalmological examinations were performed before the beginning of 
the treatment period and once in the week before early and late sacrifice. Hematology, 
coagulation, blood biochemistry and urinary investigations were performed once before the 
beginning of the treatment period, on Day 4 and at scheduled euthanasia. Blood samples for 
the determination of C-Reactive Protein (CRP) levels were taken 24 hours and 4 days after 
administration on Days 1 and 29. Blood samples for the determination of serum levels of 
antibodies against Measles antigen and Zika antigen were taken once pre-test, on Days 15 
(before administration), 31 and then 50. Blood samples for the determination of serum levels 
of antibodies against Zika antigen were also taken on Day 29 (before administration). On 
completion of the treatment (early sacrifice) or treatment-free period (late sacrifice), the 
animals were sacrificed, and a full macroscopic post-mortem examination was performed. 
Designated organs were weighed, and selected tissue specimens were preserved. A 
microscopic examination was performed on all animals.   
Results 
No unscheduled deaths occurred during the study and there were no clinical signs or local 
reactions at the injection sites. Rectal temperature and body weight were unaffected. No 
ophthalmological changes were observed at early and late sacrifice. No effects were observed 
among hematology, coagulation, blood biochemistry and urinary parameters during the study.  
CRP levels were increased 24 hours after administration on Days 1 and 29 in all test item-
treated animals and had returned to control values 4 days after the injections.  
All monkeys seroconverted to measles after the first immunization with the test item. In 
addition, it resulted in the generation of functional neutralizing antibodies against Zika virus in 
all test item treated animals and therefore confirmed the immunogenicity of the MV-ZIKA RSP 
vaccine candidate. 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 20/82 
At pathology, there were no systemic findings, but the test item administration induced mild, 
non-adverse, partially reversible inflammatory changes at the injection sites. At early sacrifice 
(Day 31), in test item-treated animals, an increased and more acute inflammation compared 
to controls was present at the injection site that was injected two days before sacrifice. It was 
characterized by infiltrates of granulocytes mainly, with increased incidence/severity of 
lymphoid infiltrates. At the injection site that was injected on Day 1, a mild inflammatory 
(granulomatous) reaction composed of macrophages and multinucleated giant cells was 
present with similar incidence and severity in both controls and test item-treated animals and 
was suggestive of an adjuvant-related reaction. None of the changes were considered to be 
adverse. At late sacrifice (Day 50), there was complete recovery of the granulocytic infiltrates 
at the injection site injected on Day 29. Minimal lymphoid infiltrates and minimal to slight 
inflammatory granulomatous reaction were still observed at the injection sites. 
Conclusion 
The potential toxicity of the test item, MV-ZIKA RSP, was evaluated in cynomolgus monkeys 
following an immunization schedule corresponding to 3 intramuscular administrations of the 
Intended Human Dose (IHD), administered every 2 weeks. Administration of the MV-ZIKA RSP 
vaccine candidate resulted in the generation of functional neutralizing antibodies against Zika 
virus in all immunized cynomolgus macaques and therefore confirmed the immunogenicity of 
the MV-ZIKA RSP vaccine candidate. The test item was well tolerated, inducing only a 
transient increase in CRP levels after administration and mild, non-adverse, partially reversible 
inflammatory changes at the injection sites. 
4.4 
Clinical Studies  
As this is the first clinical study with MV-ZIKA-RSP vaccine, no clinical data are available for 
this IMP. However, a different vaccine (MV-ZIKA) against zika have been evaluated by our 
group in one completed phase I study (MV-ZIKA-101, EudraCT 2016-004212-34). In this case, 
the MV-ZIKA vaccine tested is a recombinant live attenuated measles vaccine, based on the 
backbone of the measles Schwarz virus strain for prophylaxis of Zika infection. The 
heterologous antigens are derived from the Zika virus. 
Safety data from this study was shared in real time with the sponsor and communicated to all 
clinical trial sites in order to ensure that any safety signals that emerge are promptly addressed. 
Based on this previous clinical study, acceptable safety and tolerability as well as 
immunogenicity are expected. 
4.4.1 Phase I Study 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 21/82 
MV-ZIKA-RSP: 
No clinical study with MV-ZIKA-RSP have been performed until now. One phase 1 study with 
a similar Zika-vaccine candidate has been performed and is described below. 
 
MV-ZIKA-101 (EudraCT 2016-004212-34): 
A phase I study was conducted in two sites in Vienna, Austria. Site # 01 at the Department of 
Clinical Pharmacology and Site # 02 the Institute of Specific Prophylaxis and Tropical Medicine 
both from the Medical University Vienna. 
The name of this Phase I clinical study was “Observer blinded, randomized, placebo controlled, 
dose finding trial to evaluate the optimal dose of MV-ZIKA, a new vaccine against Zika virus, 
regarding immunogenicity, safety, and tolerability in healthy volunteers”. 
The purpose of this first-in-man study was to evaluate the optimal dose considering 
immunogenicity, safety and tolerability of the MV-ZIKA vaccine. We further investigate the 
immunogenicity, safety, and tolerability of MV-ZIKA during the treatment period up to 84 days 
after first investigational medicinal product (IMP) administration. 
All participants received study treatment on Days 0, 28, and 56. Participants randomized to 
Group A received a dose of 6x105 TCID50 MV-ZIKA on Day 0 and received placebo on Days 
28 and 56; participants in Group B received a dose of 1 x105 TCID50 MV-ZIKA on Days 0, 28, 
and 56; participants in Group C received a dose of 6x105 TCID50 MV-ZIKA on Days 0, 28, and 
56; participants in Group D (placebo) received 0.6 mL of physiological saline on Days 0, 28, 
and 56. 
Doses (volumes administered): 
Group 
Treatment 
A 
MV-ZIKA 0.6 mL (6x105 TCID50/dose) 
Placebo 0.6 mL 
B 
MV-ZIKA 0.1 mL (1x105 TCID50/dose) 
C 
MV-ZIKA 0.6 mL (6x105 TCID50/dose) 
D 
Placebo 0.6 mL 
 
The phase I clinical batch of this vaccine was present at a ready to use liquid formulation at a 
single concentration.  
The mean (SD) age of participants in the safety/mITT population was 36.8 (10.7) years and 47 
(97.9%) participants were Caucasian/White. The proportion of males (20 [41.7%]) was smaller 
than that of females (28 [58.3%]). Of the 28 females, 24 (85.7%) participants were of 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 22/82 
childbearing potential. Demographic data for the PP population were similar to the safety/mITT 
population.  
Participants were enrolled into the study from May to October 2017.  
Immunogenicity Analysis: 
The primary endpoint was immunogenicity on Study Day 56 confirmed by the presence of 
functional anti-zika antibodies as determined by a neutralizing antibody assay. An increase in 
Zika neutralizing antibody GMT was observed only in Group C (three high doses of MV-ZIKA 
[6x105 TCID50/dose]). The largest increase in neutralizing antibody GMT occurred after the 
second vaccination, increasing from 6.6 to 7.9; no increase occurred after the first vaccination 
and a reduction of 0.7 was observed after the third vaccination. No statistically significant 
difference was detected between Group C and placebo. The Group A regimen of one high 
dose of MV-ZIKA (6x105 TCID50/dose) and two placebos showed no increase in GMT. The 
Group B regimen of three low doses of MV-ZIKA showed no notable differences when 
compared with placebo (Group D). Assessment of the effect of baseline measles titer on GMT 
of neutralizing antibodies at Day 56 showed no effect. 
Seroconversions were primarily observed in participants who received high dose MV-ZIKA 
(6x105 TCID50/dose). Overall, seven participants (one in Group A, one in Group B, and five in 
Group C) showed seroconversion after MV-ZIKA vaccination and one after placebo. Variations 
in seroconversion across all treatment groups during the study did not achieve statistical 
significance. 
Participants who received the triple high-dose regimen in Group C experienced the greatest 
increase in Zika antibody GMT. The increases in GMT for Zika antibodies were notable and 
were sustained throughout the study. An increase of approximately 30% was observed after 
three high doses of MV-ZIKA, although the increase compared with placebo did not achieve 
statistical significance. Group B showed an increase in GMT for Zika antibodies after two 
vaccinations with low dose MV-ZIKA, which was of the same magnitude as the change seen 
in participants who received placebo. 
Notable increases in GMT for measles antibodies were observed in all three MV-ZIKA 
treatment groups compared with placebo but the pattern of change differed. Geometric mean 
titer increased intensely after the first vaccination with high dose MV-ZIKA, in both Group A 
and Group C; the increase in Group C was sustained and finally achieved a 1.3-fold increase 
compared with baseline after the third vaccination. In Group B, GMT for measles antibodies 
did not started to increase until after the first vaccination; the increase was sustained after the 
second vaccination and stayed at the same level after the third vaccination. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 23/82 
Due to the low number of participants showing seroconversion in the ELISA tests, it was 
decided that evaluating the cell mediated immunity would not yield additional insights and 
hence no T-cell assay was performed. 
Safety Analysis: 
MV-ZIKA was well tolerated in this population of healthy men and women. Adverse events 
were predominantly mild or moderate. Only one subject experienced a severe solicited AE 
(fever) and five participants experienced severe unsolicited TEAEs (dysmenorrhea, arthralgia, 
hyperbilirubinemia, hypercholesterolemia, hyperlipidaemia and hypertriglyceridemia); all were 
judged unlikely or not related to IMP. No AE was judged serious.  
Overall, 36 (75.0%) participants reported 200 solicited AEs and 39 (81.3%) participants 
reported 126 unsolicited AEs (TEAEs). The proportion of participants that experienced solicited 
and unsolicited AEs were higher in Group D (placebo) (100% and 87.5%, respectively) than 
Total ZIKA (70.0% and 80.0%, respectively). Only the proportion of participants that 
experienced unsolicited AEs in Group C (three high doses of MV-ZIKA [6x105 TCID50/dose]; 
100%) were higher than Group D (87.5%).  
Solicited local AEs occurred more frequently in participants who received MV-ZIKA than those 
who received placebo; more participants in Group C experienced solicited local AEs than 
Group A and Group B in all categories. The most frequently reported solicited local AE was 
injection site tenderness, followed by injection site pain. Injection site edema occurred only in 
Group C. 
Solicited systemic AEs occurred across all treatment groups. The most frequently reported AE 
was headache; although the proportion of participants who experienced headache in Group D 
(placebo) was notably higher than any MV-ZIKA treatment group (62.5% versus 12.5–43.8%). 
Adverse events of myalgia, limb discomfort, rash and fever were only reported by participants 
who received MV-ZIKA.  
The most frequently reported unsolicited AEs were nasopharyngitis and headache; the 
frequencies of these in participants who received MV-ZIKA were similar or less than those 
observed in participants who received placebo. A number of AEs were reported by participants 
who received MV-ZIKA but were not reported by those who received placebo. Most notably, 
these included general disorders and administration site conditions, and musculoskeletal and 
connective tissue disorders. Diarrhea was reported only by participants who received three 
high doses of MV-ZIKA (Group C).  
Drug-related TEAEs were reported by 18 (37.5%) participants. The proportion of participants 
who experienced drug-related TEAEs was the highest in Group C (10 [62.5%]), although Total 
ZIKA showed the same percentage (37.5%) as placebo (37.5%) and no statistically significant 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 24/82 
differences in AE rate were observed between any MV-ZIKA treatment group and placebo. 
The most frequently reported drug-related AE was diarrhea, which occurred only in Group C; 
five (10.4%) participants experienced six episodes. No subject who received single dose of 
MV-ZIKA or placebo experienced diarrhea. The second most frequently reported drug-related 
AE was vertigo, which was distributed evenly across groups (one each in Groups A, B and D). 
The proportion of participants who experienced medically attended TEAEs was higher in 
Group D (50.0%) than any MV-ZIKA treatment group (25.0–31.3%) but no statistically 
significant differences in AE rate were observed between any MV-ZIKA treatment group and 
Group D. Overall, 15 (31.3%) participants experienced medically attended TEAEs. The most 
frequently reported were arthralgia, dizziness, abdominal pain upper, hepatic steaosis and 
oropharyngeal pain, each reported by two participants. 
The proportion of participants who experienced TEAEs for which action was taken was notably 
higher in those who received placebo (87.5%) than MV-ZIKA (31.3–37.5%). There were 
statistically significant differences in AE rate in Group B versus Group D and Group C versus 
Group D. The most frequently reported AEs with action taken were nasopharyngitis (five 
[10.4%] participants) followed by headache (four [8.3%] participants). All participants who 
reported unsolicited AEs for which action was taken took medicines except one subject who 
had a broken tooth and received an amalgam filling, one subject where action taken was 
unknown and one subject who had increased liver values and was supervised by the general 
practitioner. 
No trends or changes were observed in hematology, coagulation or urinalysis. A small increase 
in mean ALT was observed in Group C. Four participants had abnormal clinical laboratory 
evaluations that were considered to be of clinical relevance but unlikely related to the study 
drug. 
No changes in vital sign parameters were considered clinically significant. No abnormalities in 
vital sign parameters were reported directly as AEs, although one subject experienced an AE 
of fever and one an AE of tachycardia. Nine participants reported 13 events of presented 
symptoms. The most frequently reported symptom was flu-like symptoms (one subject at Visit 
3 and four participants at Visit 5). One subject reported excessive fatigue. None of the 
participants reported any symptom of fever, nausea vomiting. 
More participants who received MV-ZIKA experienced local tolerability symptoms than 
placebo. The main local symptoms experienced were tenderness and injection site pain. No 
pruritus or edema was presented by any subject after injection. 
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 25/82 
4.5 
Study Rationale 
This study is designed to investigate, at first, safety and tolerability of a novel liquid vaccine 
formulation named MV-ZIKA-RSP, in healthy adults aged 18 to 55 years.  
For this purpose, two different doses of MV-ZIKA-RSP (1x105 TCID50 and 2,5 x104 TCID50 per 
dose), and one group receiving only placebo, will be assessed in healthy adults for one or two 
immunizations and at different time points. 
 
The induction of functional anti-zika-rsp antibodies by MV-ZIKA-RSP will be investigated, by 
means of immunoassays like VNT and supported with assays like ELISA and T-cell analysis. 
In our previous clinical studies, it is demonstrated that the timepoints day 28 and day 56, after 
immunisation, are suitable for elicitation of immunogenicity.  
The different vaccination schedules: 
• 
two MV-ZIKA-RSP vaccinations (high dose) on days 0 and 28  
• 
two MV-ZIKA-RSP vaccination (low dose) on day 0 and 28 
• 
 one MV-ZIKA-RSP vaccination (high dose) on day 0 and placebo on day 28 
• 
two placebo injections on day 0 and 28 
 
will allow us a comparison between immunogenicity of these dosing regimens. A single shot 
or two vaccinations within a short period of time are desirable to allow for a quick immunisation 
before travelling to areas with active transmission of the zika virus and at the same time will 
reveal valuable information on future treatment recommendations (single shot treatment vs. 
double immunisation). 
With a design of randomised, observer-blinded study we will be able to record and assess all 
adverse events. 
Since the induction of virus specific T cells is a critical step in the generation of a functional 
immune response, the rate of zika virus specific T cells will be analysed in vitro. 
This study was designed according to the Note for Guidance on Clinical Evaluation of New 
Vaccines (CHMP/VWP/164653/2005), and will be conducted in compliance with the protocol, 
Good Clinical Practice (GCP) as set forth in the International Council on Harmonisation (ICH) 
guidelines on GCP (ICH E6), and applicable local regulatory requirements.  
4.6 
Benefit/Risk Assessment 
General 
06CCVP
06G6F2

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH
7KHPHDVOHVYLUXVYHFWRULVZHOONQRZQ3RVVLEOHULVNVWKDWDUHIUHTXHQWO\DVVRFLDWHGZLWK
YDFFLQDWLRQ DUH WKH RFFXUUHQFH RI ORFDO UHDFWLRQV HJ HGHPD LQGXUDWLRQ DQG HU\WKHPD
WUDQVLHQWORFDOSDLQRUWHQGHUQHVVDWWKHLQMHFWLRQVLWHDVZHOODVPLOGWRPRGHUDWHKHDGDFKH
P\DOJLDIOXOLNHV\PSWRPVRUIDWLJXH
&XUUHQWO\QRYDFFLQHVDJDLQVW=LNDYLUXVDUHFRPPHUFLDOO\DYDLODEOHDQGIHZYDFFLQHVDUHLQ
SUHFOLQLFDORUFOLQLFDOWHVWLQJ
7KH09=,.$563YDFFLQHLVEDVHGRQWKHPHDVOHVYLUXVWHFKQRORJ\


FKHPLFDOO\V\QWKHVL]HGDQGLQVHUWHGLQWRWKH6FKZDU]YDFFLQHVWUDLQRI
PHDVOHV YLUXV 09 WR SURGXFH WKH FDQGLGDWH YDFFLQH 09=,.$563 ,PSRUWDQWO\ QR OLYH
DWWHQXDWHG =LND YLUXV LV XVHG GXULQJ JHQHUDWLRQ RU PDQXIDFWXUH RI 09=,.$563 YDFFLQH
7KXV LQGXFWLRQ RI =LND OLNH V\PSWRPV DUH QRW H[SHFWHG +RZHYHU WKHSDUWLFLSDQWV ZLOO EH
PRQLWRUHGIRURFFXUUHQFHRIDGYHUVHHYHQWVDQGWROHUDELOLW\ILQGLQJV
$VDQ\RWKHUYDFFLQH09=,.$563PLJKWLQGXFHDOOHUJLFDQGDQDSK\ODFWLFUHDFWLRQVDSDUW
IURPWKHGHVFULEHGORFDOUHDFWLRQVDWWKHYDFFLQDWLRQVLWHDQGV\VWHPLFIOXOLNHUHDFWLRQV,QUDUH
FDVHVWKHLQMHFWLRQFDQOHDGWRDYDVRYDJDOUHDFWLRQLPPHGLDWHO\DIWHULQMHFWLRQRIWKHYDFFLQH
7KHQHHGOHSULFNVIRUEORRGVDPSOLQJPD\DOVRFDXVHORFDOUHDFWLRQVVXFKDVHGHPD7KH
SDUWLFLSDQWVFDQQRWH[SHFWGLUHFWEHQHILWIURPVWXG\SDUWLFLSDWLRQH[FHSWIRUSRWHQWLDOERRVWLQJ
RIPHDVOHVLPPXQLW\DQGPRXQWLQJLPPXQLW\WR=LNDYLUXV
3RWHQWLDO%HQHILW
,IWKHFOLQLFDOGHYHORSPHQWRI09=,.$563LVVXFFHVVIXOWKLVYDFFLQHPLJKWKHOSSHRSOHZKR
DUHDWULVNRIDFTXLULQJDQLQIHFWLRQZLWK=LNDYLUXVDQGPLJKWKHOSWRSUHYHQWFRQJHQLWDOHIIHFWV
RFFXUULQJLQLQIDQWVRILQIHFWHGPRWKHUV
$ERRVWLQPHDVOHVLPPXQLW\PD\EHDQDGGLWLRQDOEHQHILW7KHLPSDFWRISUHH[LVWLQJLPPXQLW\
LVDPDMRUSRLQWRILQWHUHVWIRU0HDVOHVYHFWRUHGYDFFLQHVDVODUJHSDUWVRIWKHSRSXODWLRQKDYH
EHHQYDFFLQDWHGDWOHDVWRQFHZKLOHVRPHKDYHXQGHUJRQHQDWXUDOLQIHFWLRQV,QSUHYLRXV
SKDVH,5DPVDXHUHWDODQGSKDVH,,5HLVLQJHUHWDOFOLQLFDOWULDOVZHKDYH
VKRZQWKDWWKHLPPXQRJHQLFLW\RIDQRWKHUYDFFLQHXVLQJWKHVDPHYHFWRU09&+,.LVQRW
LQIOXHQFHGE\WKHOHYHOVRI0HDVOHVVSHFLILFVHUXP,J*EHIRUHYDFFLQDWLRQ,QDGGLWLRQZH
IRXQGWKDW WKH IUHTXHQF\ RI $GYHUVH (YHQWV$(V LQ SDUWLFLSDQWV ZLWK ORZ  L8O DQG
SDUWLFLSDQWV ZLWK KLJK  8O IUHTXHQF\ RI DQWL0HDVOHV ,J* LV VLPLODU S  ZKHQ
FRPSDULQJXQVROLFLWHG$(VS ZKHQFRPSDULQJVROLFLWHG$(V

&RQFOXVLRQ
&&93
CCI
*)

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 27/82 
Considering the small risk of adverse events and the potential benefits if immunogenicity can 
be shown, the benefit risk ratio is considered positive and the clinical study is justified. 
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 28/82 
5. 
TRIAL OBJECTIVES AND MEASUREMENTS 
5.1 
Primary Objective 
• 
To investigate safety and tolerability of MV-ZIKA-RSP, a novel liquid vaccine 
formulation, up to day 56 after first immunization consisting in one or two vaccinations 
Safety and tolerability will be assessed by the rate adverse events (AEs) after the first 
vaccination up to day 56. 
5.2 
Secondary Objectives 
• 
To investigate long-term safety, up to study day 182, after first immunization 
Long term safety and tolerability will be assessed by means of a phone call where participants 
will be asked about any adverse event (solicited, unsolicited or serious adverse event) 
experimented after the Visit 3 (day 56)  
 
• 
To investigate immunogenicity up to day 56 after first immunization consisting in one 
or two vaccinations with different doses 
 
To cover this objective the presence of functional anti-zika antibodies will be determined by 
means VNT and ELISA at the following time points: Visit 1 (day 0), Visit 2 (day 28) and Visit 3 
(day 56) after first vaccination 
• 
To get information about the optimal dose of MV-ZIKA-RSP vaccine regarding safety, 
tolerability, and immunogenicity 
An evaluation of the optimal dose will be determined after analysis of the safety, tolerability, 
and immunogenicity (VNT) 
• 
To investigate specific cell mediated immunity induced by all different treatment 
groups 
Cell mediated immunity up to day 56 will be assessed by T cell analysis of PBMC isolated on 
study days 0, 28 and 56 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 29/82 
6. 
TRIAL DESIGN 
6.1 
Overall Study Design 
This is an observer-blinded, block-randomised, dose finding, phase I trial, comparing different 
dose levels of MV-ZIKA-RSP to evaluate safety, tolerability and immunogenicity, of this novel 
ZIKA-RSP vaccine. Placebo (physiological saline solution) will be applied to blind the different 
treatment schedules. 
The study will be conducted at a single study site, Medical University Vienna and is registered 
online at http://www.ClinicalTrials.gov under EudraCT number 2019-000840-93. 
After the screening procedures, 48 healthy male and female volunteers aged 18-55 years will 
be randomly assigned to one of four treatment groups (A, B, C or D). Participants will be 
assessed for immunogenicity on days 0, 28 and 56 (treatment period), as confirmed by the 
presence of functional anti-zika-rsp antibodies determined by (VNT) and by ELISA, at the same 
time safety will be also assessed. After the treatment period, participants will be call by phone 
(day 182) for evaluation of safety follow-up.  
The investigator and site personnel assessing AEs, all participants, as well as one of the 
sponsor’s representatives involved in the monitoring and conduct of the study will be blinded 
to which vaccine was administered. Only the unblinded monitor, site personnel performing 
randomisation and preparation of IMP will be unblinded. 
Study duration per subject will be 182 days (6 months), consisting of 4 weeks treatment period 
(study day 0 till study day 28) and 22 weeks follow up period (study day 2 till study day 182) 
6.2 
Study Endpoints 
6.2.1 Primary Endpoint 
• 
Rate of adverse events (AEs) up to study day 56 
6.2.2 Secondary Endpoints 
• 
Rate of solicited and unsolicited AEs, as well as, reported serious adverse events 
(SAEs) up to study day 182 (long-term safety) compared between the 4 different 
treatment groups.  
• 
Immunogenicity on study days up to study day 56 as confirmed by the presence of 
functional anti-zika antibodies as determined by VNT and by ELISA 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 30/82 
• 
Cell-mediated immunity specific for ZIKA-RSP, up to study day 56, confirmed by the 
presence of specific functional CD4+ and CD8+ T-cells 
• 
Safety laboratory parameters (haematology, serum chemistry, urinalysis) 
6.3 
Treatment Groups 
After the screening procedure, 48 participants eligible for the study will be randomised to one 
of the following 4 treatment groups: 
Group A:  
two vaccinations with MV-ZIKA-RSP, high dose on day 0 and day 28 
Group B:  
two vaccinations with MV-ZIKA-RSP, low dose on day 0 and day 28 
Group C:  
one vaccination with MV-ZIKA-RSP, high dose, on day 0 and placebo on day 28 
Group D: 
placebo on day 0 and day 28 
 
Table 1. Treatment groups and vaccination schedule 
 
Group 
Treatment 
Dose 
Schedule 
Participants 
A 
MV-ZIKA-RSP high 
dose 
 
1 x105 TCID50 
days 0 & 28 
14 
B 
MV-ZIKA-RSP low 
dose 
2,5 x104 TCID50 
 
 
days 0 & 28 
14 
C 
MV-ZIKA-RSP high 
dose 
+ 
Placebo 
1 x105 TCID50 
+ 
Placebo 
MV-ZIKA-RSP high dose 
day 0  
+ 
Placebo day 28 
12 
D 
Placebo 
 
Placebo 
Placebo day 0  
+ 
Placebo day 28 
8 
 
The actual vaccine dosages are dependent on the manufacturing process, specifying and 
allowing a window of 0.5 log.  
 
 
 
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 31/82 
6.4 
Study Flowchart 
 
 
 
 
 
 
 
 
 
6.5 
Study Procedures 
 
Table 2. Study procedures 
 
Screenin
g 
 
Treatment 
 
Post Treatment 
Safety Follow 
Up 
ET 
Visit 
VS 
V1 
Post 
dose 
safety FU 
V2 
V3 
 Primary 
Endpoint 
V4 
(1) 
Study Day 
-14 to -1 
0 
 
28±
3 
56 ±3 
182 ±14 
 
Informed Consent 
x 
 
 
 
 
 
 
Inclusion/Exclusion Criteria 
x 
x 
 
 
 
 
 
Medical History 
x 
x 
 
 
 
 
 
Vaccination History  
x 
x 
 
 
 
 
 
Vital Signs (2) 
x 
x 
 
x 
x 
 
x 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 32/82 
Symptom-directed Physical Examination 
 
x 
 
x 
x 
 
x 
System-based Physical Examination (6) 
x 
x  
 
x 
x  
 
x  
Adverse Events (3) (7) 
 
x  
x 
x 
x 
x 
x 
Prior/Concomitant Medication (4) 
x 
x 
x 
x 
x 
x 
x 
Clinical Laboratory (5) 
x 
 
 
x 
x 
 
x 
Randomization 
 
x 
 
 
 
 
 
Urine Pregnancy Test (12) 
x 
x 
 
x 
x 
 
x 
Study Treatment  
 
x 
 
x 
 
 
 
Immunogenicity (VNT) (12) 
 
x 
 
x 
x 
 
x 
T-cell immunity (PBMCs) (12) 
 
x 
 
x 
x 
 
x (11) 
Zika antibody (ELISA) (12) 
 
x 
 
x 
x 
 
x 
Measles antibody (ELISA) (12) 
 
x 
 
x 
x 
 
x 
Local Tolerability (7) 
 
x 
 
x 
x 
 
x 
Dispense subject diary 
 
x 
 
x 
 
 
 
Collect and review subject diary 
 
 
 
x 
x 
 
x 
Safety phone call (9) 
 
 
x (9) 
 
 
x (10) 
 
 
(1) 
Early termination visit should – if possible - be conducted 28±3 days after last IMP administration. 
(2) 
Systolic/diastolic blood pressure, pulse rate, body temperature 
(3) 
Conditions/symptoms noted prior to randomization should not be reported as AE, but should be reported as medical 
history 
(4) 
Medications administered up to 30 days prior to screening until end of study participation should be documented 
(5)  
The clinical laboratory comprises: Hematology (hemoglobin, hematocrit, erythrocyte count, differential white blood count, 
platelets), Chemistry (creatinine, potassium, sodium, calcium, AST, ALT, alkaline phosphatase, bilirubin), Coagulation 
(prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen), Urinalysis (standard urine test stick for 
determining pH-value, glucose, protein, bilirubin, urobilinogen, red blood cells, white blood cells, nitrite, ketone and specific 
gravity), Virology (only at screening; HBs-Ag, anti-HBc-Ab, anti-HCV-Ab, HIV 1/2 Ab/HIV-1 Antigens if not done within 30 
days before screening) 
(6) 
To be performed if necessary, according to findings in symptom-directed physical examination 
(7) 
To be performed 1 hour after each IMP administration and additionally at the subsequent visit (prior to IMP administration, 
if applicable) 
(8) 
Participants should be interviewed regarding potential AEs since the last study visit. If indicated, participants should be 
called in for an unscheduled visit. 
(9) 
The post dose safety call follows up should be conducted 7±1 days (one week) after the first vaccination. 
(10) 
The post treatment safety call follows up should be conducted 182 days (6 months) after the first study treatment also in 
case of an early termination. 
(11) 
The amount of blood sample taken during the early termination (ET) visit depends on the time point of the visit. If the ET-
visit replaces the primary endpoint visit on day 56±3, 48 to 60 mL blood should be taken; otherwise 24 to 48 mL are 
enough. 
(12) 
Urine for pregnancy test should be taken before vaccination 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 33/82 
7. 
PARTICIPANT IDENTIFICATION 
7.1 
Trial Participants 
Healthy male and female volunteers will be screened for inclusion and exclusion criteria and 
randomised to one of 4 treatment groups in case of eligibility.  
Forty-eight participants will be enrolled if all the following inclusion criteria and none of the 
following exclusion criteria apply: 
7.2 
Inclusion Criteria 
1. 
Signed informed consent obtained before any trial-related activities 
2. 
Healthy men or women aged 18 to 55 years on the day of consenting 
3. 
Ability to comprehend the full nature and purpose of the study, including possible risks and 
side effects; ability to cooperate with the investigator and to comply with the requirements 
of the entire study 
4. 
All female participants must have a negative urine pregnancy test at screening  
5. 
Willingness not to become pregnant or to father a child during the entire study period by 
practicing reliable methods of contraception as specified in protocol section 8.11.4 
6. 
Availability during the duration of the trial 
7. 
Normal findings in medical history and physical examination or the investigator considers 
all abnormalities to be clinically irrelevant 
8. 
Normal laboratory values or the investigator considers all abnormalities to be clinically 
irrelevant (unless otherwise specified in exclusion criteria) 
7.3 
Exclusion Criteria 
The subject may not be enrolled in the study if any of the following applies: 
1. Participation in another clinical study (including exposure to an investigational medicinal 
product ore device) within one month before the screening visit or planned concurrent 
participation in another clinical study before completion of the treatment period (day 56) 
2. History of immunodeficiency, known human immunodeficiency virus (HIV) infection or 
current hepatitis B/C infection 
3. Strong anamnestic evidence for or confirmed history of or current infection with Zika- or 
Dengue-virus 
4. History of drug addiction including alcohol dependence within the last 2 years  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 34/82 
5. Inability or unwillingness to avoid intake of more than around 20g alcohol per day during 
48 hours after each vaccination (equals roughly 0.5 L beer or 0.25 L of wine) 
6. Vaccination within 4 weeks prior to first vaccination or planning to receive any non-study 
vaccine until end of treatment period (day 56)  
7. Prior receipt of any Zika or Chikungunya vaccine 
8. History of moderate or severe arthritis or arthralgia within the past 3 months prior to 
Screening Visit 
9. Recent infection within 1 week prior to Screening Visit (possibility of deferral) 
10. Blood donations including plasma donations, 90 days prior to Screening Visit and 
anticipated blood, plasma, tissue, sperm, or organ donation, throughout the study until end 
of treatment period (day 56) 
11. Clinically relevant history of renal, hepatic, gastrointestinal, cardiovascular, respiratory, 
skin, haematological, endocrine, inflammatory, autoimmune, or neurological diseases or 
clinically relevant abnormal laboratory values, that in the opinion of the investigator may 
interfere with the aim of the study 
12. History of neoplastic disease (excluding non-melanoma skin cancer that was successfully 
treated) within the past 5 years or a history of any haematological malignancy 
13. Behavioural, cognitive, or psychiatric condition that in the opinion of the investigator affects 
the ability of the participant to understand and cooperate with the study protocol 
14. History of severe adverse reactions to vaccine administration, including anaphylaxis and 
related symptoms, such as urticaria, respiratory difficulty, angioedema and abdominal pain 
to vaccines, or history of allergic reaction likely to be exacerbated by any component of 
the vaccine 
15. History of anaphylaxis to drugs or other allergic reactions, which the investigator considers 
compromising the safety of the volunteer 
16. Abnormal laboratory values which, at the discretion of the investigator should lead to the 
exclusion of the subject 
17. Use of medication during 2 weeks before the first vaccination and throughout the study, 
which the investigator considers affecting the validity of the study, except hormonal 
contraception or hormonal replacement therapy in female participants. (Prior to taking any 
medication within 72 h before study vaccination, the subject should consult the 
investigator) 
18. Use of immunosuppressive drugs like corticosteroids (excluding topical preparations) 
within 30 days prior to first IMP administration, or anticipated use until completion of the 
end of treatment visit Receipt of blood products or immunoglobulins within 120 days prior 
to the Screening Visit or anticipated receipt of any blood product or immunoglobulin before 
completion of the treatment period (day 56) 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 35/82 
19. Pregnancy (positive pregnancy test at screening or during the treatment period) or 
lactation at screening, or planning to become pregnant during the treatment period 
20. Unreliable contraception methods (for details please refer to protocol section 8.11.4) 
21. Persons in a direct relationship with the sponsor, an investigator or other study team 
members. Direct dependent relationships include close relatives (i.e. children, parents, 
partner/spouse, siblings) as well as employees of the study site or the sponsor 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 36/82 
8. 
TRIAL PROCEDURES 
8.1 
Informed Consent (AMG § 38) 
In obtaining and documenting informed consent, the investigator must comply with the 
applicable regulatory requirement(s) and adhere to ICH guideline for GCP, the requirements 
in the Declaration of Helsinki and the EU directive 2005/28/EC. 
Before any trial specific procedures are performed participants will be informed about the exact 
nature of the trial, what it will involve for the participant, all procedures as determined by the 
clinical protocol, the known side effects and any risks involved in taking part. It will be clearly 
stated that the subject is free to withdraw from the trial at any time for any reason without 
prejudice to future care, without affecting their legal rights and with no obligation to give the 
reason for withdrawal. 
The participant will be allowed as much time as needed to consider the information, and the 
opportunity to question the investigator or other independent parties to decide whether they 
will participate in the trial. 
Written informed consent will then be obtained by means of participant’s dated signature and 
dated signature of the qualified and experienced person who presented and obtained the 
informed consent. A copy of the signed informed consent will be provided to the subject and 
the original signed form will be retained at the site.  
If information becomes available that may be relevant to the subject’s willingness to continue 
participating in the trial, the investigator must inform the subject in a timely manner, and a 
revised written informed consent must be obtained. 
8.2 
Randomisation 
At Visit 1 (day 0) 48 eligible participants will be randomly assigned to one of four treatment 
groups (A, B, C or D) by means of randomisation envelopes provided by data management.  
Each subject will be assigned a unique three-digit randomisation number in ascending order, 
by opening the randomisation envelope with the lowest free (unassigned) randomisation 
number available. Randomisation should be performed as late as possible, i.e. when 
knowledge of the assigned treatment group becomes necessary. Only trained members of the 
unblinded study team will be authorised to open one envelope per subject, containing the 
information about the allocated treatment group. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 37/82 
Upon opening, the randomisation envelope shall be signed with date and time of randomisation 
and the corresponding subject number. 
Please note:  
As this is a first in humans’ study, the site should initially enroll a group of six participants and 
then pause with enrollment until these participants completed their Post Dose Safety Follow 
Up one week after the first vaccination. If no events as defined in section 8.9.1 "Safety Stopping 
Rule" are identified, the site should resume enrolling participants. If such events were 
identified, the investigators and the sponsor need to assess the risk of continuing the study 
and based on this assessment to decide if or if not, the study should be continued. 
The 48 participants will be distributed between the 4 groups using a randomisation ratio of 14 
: 14 : 12 : 8 = A : B : C : D, with an adequate representation of high dose, low dose and placebo 
subjects (2 : 2 : 0 : 2 = A : B : C : D) within the first 6 subjects to allow for the safety evaluation 
after the first vaccination. 
A subject will be considered enrolled once the first IMP administration has been performed 
according to randomisation. 
8.3 
Blinding and Code-breaking  
As this study will be conducted in an observer-blinded manner, unblinded study team members 
responsible for randomisation, monitoring (unblinded monitor) and preparation of IMP, will 
otherwise not be involved in the conduct of the study. 
All participants, the investigator and site personnel performing study related assessments, as 
well as the sponsor’s representatives involved in the monitoring (blinded monitor) and conduct 
of the study, will be blinded to treatment assignment. 
Besides one set of randomisation envelopes, the site will receive an additional set of 
emergency envelopes stored in the ISF for unblinding purposes. As the knowledge of the 
subject’s treatment might become mandatory (e.g. in case of emergency) the investigator has 
the possibility to open the emergency envelope identified with the subject’s randomisation 
number and unblind the subject.  
Please note that this code-breaking should only be done by the investigator if the knowledge 
of the subject’s treatment influences the decision on further procedures. If unblinding would 
not make any difference for further treatment, the study team should remain blinded. 
8.4 
Screening Procedures 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 38/82 
8.4.1 Screening Visit (VS, 1-14 days prior to Visit 1)  
After participants have signed the informed consent form, the screening visit will be performed 
within two weeks prior to Visit 1. 
During this visit, all participants will be checked for inclusion and exclusion criteria, undergo a 
full physical examination and evaluation of vital signs (systolic and diastolic blood pressure, 
pulse rate and body temperature). Medical history, vaccination history (covering the last three 
years prior to screening), prior/concomitant medication within 30 days prior to screening and 
demographic data will be recorded. 
Blood and urine samples will be collected for clinical laboratory assays (HIV, hepatitis B and 
C, haematology, coagulation parameters, clinical chemistry and urinalysis). HIV tests with 
available results performed up to 30 days before screening are acceptable. A urine pregnancy 
test will be performed in women of childbearing potential. For details on laboratory sampling 
and testing, see section 8.11.1.  
All participants eligible for the clinical study will be asked to return to the study site within 14 
days after the screening visit for Visit 1. 
8.5 
Treatment Period 
8.5.1 Visit 1 (day 0) 
Participants found to be eligible will return to site for Visit 1 within 14 days after the screening 
visit. Inclusion and exclusion criteria will be checked again and (in case of changes since the 
screening visit) the medical history will be updated. Symptom-directed physical examination, 
system-based assessment (only if necessary, according to findings in symptom-directed 
physical exam) and vital signs (systolic and diastolic blood pressure, pulse and body 
temperature) will be recorded. In addition, the participants will be asked regarding changes in 
concomitant medications since the last visit.  
Blood samples for baseline immunogenicity assessments (Zika antibody ELISA, VNT) will be 
drawn from an appropriate forearm or cubital vein. For T-cell assays blood samples will be 
collected and peripheral blood mononuclear cells (PBMCs) will be prepared. A urine pregnancy 
test will be performed in women of childbearing potential. For details on laboratory sampling 
and testing, see section 8.11. 
Participants will be randomised to treatment group A, B, C or D and receive either the first 
MV-ZIKA-RSP vaccination or placebo, according the assigned randomized treatment group. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 39/82 
The participants’ diaries will be explained and handed over. Participants will be asked to report 
their body temperature and any adverse event in these diaries for 7 days, starting 6 hours after 
the injection. 
After observation of the participants for 1-hour, local reactions and adverse events (if any 
occurred) will be recorded.  
Blood pressure and pulse rate will be checked, and participants will be discharged only if the 
investigating physician considers it to be safe for them to leave the department.  
The participants are invited to return to study site for Visit 2 on study day 28 (± 3 days). 
8.5.2 Post Dose Safety Follow-up (day 7 1 days) 
The investigator will ensure phone contact with the participants one week after first vaccination 
was performed and ask about any new concomitant medications or AEs occurred since the 
last visit. If deemed necessary, the investigator may ask participants to come to study site for 
an unscheduled visit to assess severity of the AEs. 
8.5.3 Visit 2 (day 28 3 days) 
The investigator or designee will examine local tolerability (of previous vaccination site) and 
ask about any new concomitant medications or AEs occurred since the last visit.  
In addition, symptom-directed physical examination, system-based assessment (only if 
necessary, according to findings in symptom-directed physical exam) and vital signs (systolic 
and diastolic blood pressure, pulse rate and body temperature) will be recorded.  
A urine pregnancy test will be performed in women of childbearing potential. 
Blood samples will be taken for clinical chemistry, haematology, and coagulation parameters. 
Additional blood samples for immunogenicity assessments (Zika and Chikungunya ELISA, 
VNT) will be drawn from an appropriate forearm or cubital vein. For T-cell assays blood 
samples will be collected and PBMCs will be prepared. For details on laboratory sampling and 
testing, see section 8.11. 
Participants will receive their second vaccination either with MV-ZIKA-RSP, or placebo, 
according to the assigned randomised treatment group. 
The participants’ diaries will be explained and handed over. Participants will be asked to report 
their body temperature and any adverse event in these diaries for 7 days, starting 6 hours after 
the injection.  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 40/82 
After observation of the participants for 1-hour, local reactions, and adverse events (if any 
occurred) will be recorded.  
Blood pressure and pulse rate will be measured, and participants will be discharged only if the 
investigating physician considers it to be safe for them to leave the department.  
The participants are invited to return to study site for Visit 3 on study day 56 (± 3 days). 
8.5.4 Visit 3 (day 56 3 days) 
The investigator or designee will examine and document local tolerability (of the previous 
vaccination site) and ask about any new concomitant medications or AEs occurred since the 
last visit. 
Symptom-directed physical examination, system-based assessment (only if necessary, 
according to findings in symptom-directed physical exam) and vital signs (systolic and diastolic 
blood pressure, pulse rate and body temperature) will be recorded.  
A urine pregnancy test will be performed in women of childbearing potential. 
Blood samples will be taken for clinical chemistry, haematology, coagulation parameters and 
urine samples will be collected for urinalysis. 
Additionally, blood samples for immunogenicity assessments (Zika and Chikungunya ELISA, 
VNT). For T-cell assays blood samples will be collected and PBMCs will be prepared. For 
details on laboratory sampling and testing, see section 8.11. 
Upon Visit 3, the treatment period of this study will be completed.  
Participants are informed about the approximated date for a safety follow up call for follow up 
Visit 4 on study day 182 (± 14 days).  
8.6 
Post Treatment Safety Follow up Period 
8.6.1 Visit 4 (day 182 14 days) 
Participants will be call and will be ask if they experienced any new or changes in adverse 
events (AE) or concomitant medications since the last visit.  
8.7 
Early Termination Visit (ET) 
Participants who are withdrawn or wish to discontinue the study prematurely for any reason 
shall be asked to attend an early termination visit.  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 41/82 
If the early termination occurs within the treatment period (at or before Visit 4), participants 
shall perform all procedures as defined for Visit 3. If the early termination occurs within the 
follow up period (after Visit 3), participants shall perform all procedures defined for Visit 4. 
8.8 
Unscheduled Visits 
Unscheduled visits can be conducted whenever deemed necessary. The extent of the 
examination performed at an unscheduled visit is at the discretion of the investigator. All 
procedures completed during an unscheduled visit should be documented in the source data 
and should also be entered to the electronic case report form (eCRF). 
8.9 
Discontinuation/Withdrawal of Participants 
Participants may withdraw from participation in the study at any time, without the need to justify.  
If a subject withdraws or is withdrawn prior to completion of the study, the reason for this 
decision should be recorded in the electronic case report forms (eCRFs) although it is 
recognised that the subject may refuse to provide a reason.  
Generally, any enrolled/vaccinated subject may withdraw or be withdrawn from the study for 
the following reasons: 
• 
The subject withdraws consent 
• 
The subject fails to comply with the requirements of the protocol 
• 
At the discretion of the investigator 
• 
For an adverse event, which at the discretion of the investigator requires discontinuation 
of the subject or results in inability to continue to comply with trial procedures 
• 
if the administration of a drug, which is not permitted by the study protocol, becomes 
necessary (this should be discussed and agreed between the investigator and the sponsor) 
• 
violation of exclusion criteria 1, 4, 5, 7, 12, 15, 16, 17, 18, 19 or 20 during the study (this 
should be discussed and agreed between the investigator and the sponsor) 
All participants who withdraw or are withdrawn prematurely from the study will undergo an 
early termination visit (see section 8.7). 
8.9.1 Safety Stopping Rules 
Stopping rules are established at subject level and entire study level. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 42/82 
Individual Participants Stopping Rules 
The following are specific criteria for discontinuing individual participants from further 
vaccination, but not from completing scheduled follow-up assessments, unless the participant 
is explicitly withdrawn from the study: 
- Participants who develop serious adverse reaction (SAR) at the discretion of the 
investigator 
- Participants who develop a potentially life-threatening (grade 4) solicited systemic 
adverse reaction that occurs without alternative etiology within 7 days after the study 
vaccination 
- Participants who develop a medical condition for which continued participation, in the 
opinion of the investigator, would pose a risk to the subject or would be likely to 
confound interpretation of the results 
- Participants who become pregnant during the treatment phase of the study 
- Participants who experience anaphylaxis within 24 hours or generalised urticaria within 
72 hours after administration of the study vaccine  
If any of these occur, appropriate measures to treat the subject will be taken and the sponsor 
or the sponsor designee will be notified immediately. 
Entire Study Stopping Rules 
The study will be stopped (no new enrolments and no additional vaccine administered) pending 
a safety review by the investigator and the sponsor, if any of the following occurs: 
- One or more participants experience a serious adverse reaction (SAR), based on the 
current reference safety information (Investigator’s Brochure) 
- One or more participants’ deaths assessed by the investigator, safety manager or 
sponsor as related to investigational product 
- One or more participants develop laryngospasm, bronchospasm or anaphylaxis 
within 24 hours of vaccine administration 
- Two or more participants develop injection-site ulceration, abscess or necrosis 
- Two or more participants experience generalised urticaria within 72 hours after 
administration of study product 
Upon completion of this review, the sponsor will determine if study enrolment or treatment 
should be discontinued, or if study enrolment and treatment may continue according to the 
protocol.  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 43/82 
Irrespective of the stopping rules the study may be temporarily suspended or prematurely 
terminated at any time if there is sufficient reasonable cause, and if agreed to by both the 
investigator and the sponsor as being in the best interests of participants and justified on either 
medical or ethical grounds. In terminating the study, the sponsor, the CRO, and the investigator 
will ensure that adequate consideration is given to the protection of the participants’ interests. 
Whenever the study is stopped the competent authorities and ethical committees will be 
informed. 
In case the entire study has to be discontinued prematurely, all concerned participants should 
be asked to perform an early termination visit. 
8.9.2 Replacement of Participants 
Participants who are discontinued before randomisation will be regarded as screening failures 
and screening will be continued until 48 participants will be randomised. 
Participants who withdraw or are withdrawn after randomisation will not be replaced. This 
may result in a decreased number of participants in the predefined sample size. 
8.10 End of Study 
The end of study will be defined as the date of the last visit of the last subject. The study will 
then be stopped. For premature termination or suspension of the study, refer to section 8.9.1 
8.11 Handling of Samples 
The procedures of sample collection, preparation and storage will be described in detail in a 
study specific Lab-Manual. Blood sampling kits and labels will be supplied by the supply 
vendor. Timepoints and blood volumes are summarised in Table 3. 
8.11.1 
Samples for Safety Laboratory Parameters 
Safety laboratory parameters will be analysed by local laboratories according to applicable 
laboratory SOP. 
The following parameters will be measured to evaluate inclusion and exclusion criteria only 
once at screening visit (SV).  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 44/82 
- Virology: Hepatitis B surface-Ag, anti-Hepatitis B core-Ab, anti-Hepatitis C-Ab, HIV Ab. HIV 
tests that were performed up to 30 days before screening and where results are available 
are acceptable and do not require a new test. 
The following laboratory parameters will be assessed for subject’s safety at screening (VS), 
Visit 2, Visit 3 and additionally at Early Termination (if applicable) as depicted in Table 2. 
- Haematology: haemoglobin, haematocrit, erythrocyte count, differential and total white 
blood count, platelets 
- Chemistry: creatinine, potassium, sodium, calcium, AST, ALT, alkaline phosphatase, 
bilirubin 
- Coagulation parameter: prothrombin time (PT), activated partial thromboplastin time 
(aPTT), fibrinogen  
- Urinalysis: a standard urine test dipstick for determining pH-value, glucose, protein, 
bilirubin, urobilinogen, blood, leucocytes esterase, nitrite, ketone and specific gravity will 
be used.  
- Laboratory values deviating from the normal ranges defined by the local laboratory will be 
evaluated by the investigator. Clinically relevant deviations have to be reported as adverse 
events and graded according to the FDA Guidance for Industry (Toxicity Grading Scale for 
Healthy Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical Trials, 
Sept. 2007) 
8.11.2 
Immunogenicity Samples 
Virus Neutralization Test (VNT) 
Humoral systemic immune response will be determined by anti-zika-rsp virus neutralisation 
test. The virus neutralisation titre (VNT) will be measured to identify immunogenicity in sera of 
all participants. 
At Visits 1, 2, 3 and ET (if applicable) a volume of 8ml blood will be collected, to isolate serum 
and freeze a minimum of three aliquots (0.5 ml /vial) at -80°C. 
Enzyme linked immunosorbent assay (ELISA) 
In addition, humoral systemic immune response mediated by serum IgG antibodies against 
zika-rsp will be determined by IgG ELISA.  
At Visits 1, 2, 3 and ET (if applicable) a volume of 8 ml blood will be collected, to isolate serum 
and freeze a minimum of three aliquots (1 ml /round bottom cryovial) at -80°C for determination 
of anti-zika antibodies by ELISA. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 45/82 
Anti-measles antibodies will be determined by ELISA out of the same samples collected at 
Visits 1, 2, 3 and ET (if applicable). Therefor no additional blood sapling will be required for 
this assay. 
T cell Analysis 
Cellular immunogenicity will be determined by evaluation of T cell immune response. 
Therefore, PBMCs will be isolated from 24-48 ml of fresh collected blood within 8 hours after 
blood draw, aliquoted in approximately 3-5 cryovials (1 ml /vial), to an approximated 
concentration of 10x 106 cells per vial, frozen at -80°C, and storage in N2, for further analysis. 
The T cells will be re-stimulated in vitro with zika virus structural protein peptides to determine 
the number of zika-virus specific T cells after one or two immunisations. 
At Visits 1, 2, 3 and ET (if applicable) a volume of 24-48 ml blood will be collected for isolation 
of peripheral blood mononuclear cells (PBMCs).  
 
 
 
 
Table 3. Blood sample collection 
Test 
Type of Vials 
Volume 
ml 
Visits 
Total 
Volume ml 
Virology (HIV, HBV, HCV) 
serum 
* 0  
VS, V2, V3 
* 0 
Haematology 
EDTA 
4 - 5  
VS, V2, V3 
12-15 
Coagulation 
citrate tubes 
4 - 5  
VS, V2, V3 
12-15 
Clinical Chemistry 
serum 
8  
VS, V2, V3 
24 
VNT 
serum 
8  
V1, V2, V3 
24 
ELISA ZIKA 
serum 
8  
V1, V2, V3 
24 
ELISA measles 
serum 
** 0 
V1, V2, V3 
** 0 
PBMCs for T-cell Analysis  
citrate tubes 
24-48 
V1, V2, V3 
96 
 
 
 
 
192-198 
 
 
* 0 ml: Virology testing at screening, V2 and V3 will be performed from the same sample as Clinical Chemistry, no 
additional blood draw will be necessary 
** 0 ml: ELISA for determination of measles antibodies will be performed from the same sample as the ELISA for 
ZIKA-Ab, no additional blood draw will be necessary 
 
Total blood volume collected during the study duration per subject (182 days/6 months) will 
be approximately –192-198 ml. 
8.11.3 
Pregnancy Test – Contraception 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 46/82 
For females of childbearing potential, urine pregnancy tests will be performed at all scheduled 
visits and during the V4-phone call, participants will be ask whether they are pregnant. Only 
females with a negative urine test will be allowed to receive study vaccinations. 
At visits where IMP is to be administered, a negative urine test outcome must be available prior 
to administration of study treatment. 
Participants with a positive urine pregnancy test at Screening Visit cannot be enrolled into the 
study. In case of pregnancy during the study the investigator will inform the sponsor 
immediately by completing and sending the applicable form (Section 10.6). The participant 
should attend planned visits without further vaccinations for follow-up. If at least one dose was 
received, participants will be monitored until pregnancy outcome. 
 
Contraception 
Birth control methods which may be considered as highly effective are those that can achieve 
a failure rate of less than 1% per year when used consistently and correctly are considered as 
highly effective birth control methods. Such methods include: 
_ combined (estrogen and progestogen containing) hormonal contraception associated with 
inhibition of ovulation: 
• 
Oral 
• 
Intravaginal 
• 
transdermal 
_ progestogen-only hormonal contraception associated with inhibition of ovulation: 
• 
oral 
• 
injectable 
• 
implantable  
_ intrauterine device (IUD) 
_ intrauterine hormone-releasing system (IUS) 
_ bilateral tubal occlusion  
_ vasectomised partner 
_ sexual abstinence  
 
Acceptable birth control methods which may not be considered as highly effective are those 
that result in a failure rate of more than 1% per year include: 
_ progestogen-only oral hormonal contraception, where inhibition of ovulation is not the 
primary mode of action 
_ male or female condom with or without spermicide  
_ cap, diaphragm, or sponge with spermicide 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 47/82 
 
Birth control methods which are considered unacceptable include:  
Periodic abstinence (calendar, symptothermal, post-ovulation methods), withdrawal (coitus 
interruptus), spermicides only, and lactational amenorrhoea method (LAM) are not acceptable 
methods of contraception. Female condom and male condom should not be used together 
8.11.4 
Labelling, Storage and Transport of Samples  
Please refer to the study specific Lab-Manual for detailed information about labelling, storage 
and transport of study serum samples vials and blood for PBMC isolation. 
Labelling of Samples 
Each sample tube will be clearly and unequivocally identified by a label resistant to the storage 
temperature which contains at least the following information:  
study code, subject number, visit number, test allocation (e.g. ELISA-ZIKA, ELISA-measles, 
VNT) 
Samples Storage  
Aliquots of samples will be stored at -80°C in a temperature monitored freezer. The 
temperature will be controlled either by a connected central monitoring system or a min/max 
thermometer, which will be controlled and documented each working day.  
Handling and storage of blood for PMBC preparation will be explained in a corresponding 
section in the above-mentioned Lab-Manual. 
 Transport of Samples  
Study samples packed in sufficient dry ice will be shipped by an authorised courier from the 
study site either to the sponsor, a dedicated analytical laboratory or a sample storage facility. 
Pick up of serum samples will be organised together with the site personnel at the end of the 
study or during the study as necessary. 
 
06CCVP
06G6F2

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH

,19(67,*$7,21$/0(',&,1$/352'8&7,03

,QYHVWLJDWLRQDO0HGLFLQDO3URGXFW'HVFULSWLRQ
 09=,.$563YDFFLQHWHVWSURGXFW
7KH09=,.$563YDFFLQHFDQGLGDWHLVDOLYHDWWHQXDWHGUHFRPELQDQWYLUDOYHFWRUHGYDFFLQH
IRUSURSK\OD[LVRI=LNDYLUXVLQIHFWLRQ

FKHPLFDOO\
V\QWKHVL]HGDQGLQVHUWHGLQWRWKH6FKZDU]YDFFLQHVWUDLQRIPHDVOHVYLUXV09WRSURGXFHWKH
FDQGLGDWHYDFFLQH09=,.$563IXUWKHUGHWDLOVXQGHUVHFWLRQDOVRUHIHUWR,%DQG,03'
7KH09=,.$563YDFFLQHLVSURYLGHGE\WKHVSRQVRULQJODVVYLDOV
7KHVWXG\GUXJXVHGLQWKHVWXG\LVDOLTXLGIUR]HQVXVSHQVLRQIRULQMHFWLRQZLWKDQH[WUDFWDEOH
GRVHRI
+LJKGRVH&RQFHQWUDWLRQ[7&,'SHUYLDO
/RZGRVH&RQFHQWUDWLRQ[7&,'SHUYLDO
7ZRGRVDJHVZLOOEHDGPLQLVWHUHGWRWKHSDUWLFLSDQWV
+LJKGRVH&RQFHQWUDWLRQP/[7&,'GRVH
/RZGRVH&RQFHQWUDWLRQP/[7&,'GRVH
'RVDJHVWUHQJWKVKDYHEHHQVHOHFWHGEDVHGRQH[SHULHQFHZLWKWKLVW\SHRIYDFFLQH6FKZDU]
VWUDLQPHDVOHVYLUXVYHFWRUHGYDFFLQH
/LTXLGIUR]HQ09=,.$563YDFFLQHZLOOEHWKDZHGDWURRPWHPSHUDWXUHIRUDERXWPLQ
P/ZLOOEHGUDZQXSLQWRDP/V\ULQJH7KHSUHSDUHUPD\LQMHFWWKHVXEMHFWRUFDQKDQG
RYHUWKHV\ULQJHWRDQLQMHFWRU7KHLQMHFWLQJSUHSDUHUPXVWQRWSDUWLFLSDWHLQWKHVWXG\DV
REVHUYHUWKHLQMHFWRULISDUWLFLSDWLQJLQODWHUVWXG\FRQGXFWPXVWEHEOLQGHGHJE\EOLQGLQJ
WKHFRQWHQWRIWKHV\ULQJHZLWKWKHSURYLGHGV\ULQJHODEHO
 3ODFHER
$ VWHULOH SK\VLRORJLFDO VDOLQH VROXWLRQ ZLOO EH XVHG DV SODFHER WR HQVXUH EOLQGLQJ RI WKH
WUHDWPHQWVFKHGXOHLQFDVHRIVLQJOHYDFFLQDWLRQUHJLPHQWUHDWPHQWJURXS&DQG'7KH
SK\VLRORJLFDOVDOLQHVROXWLRQ1D&OLVDFOHDUFRORXUOHVVVROXWLRQUHDG\WRXVHVWRUHGDW
URRPWHPSHUDWXUHDQGSURYLGHGE\WKHVWXG\VLWH

&&93
CCI
*)

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 49/82 
9.2 
Packaging and Labelling 
Individual cardboard boxes will contain single 2 ml transparent glass vials, with rubber stopper 
and aluminium seal, containing frozen liquid. The labels on vials and boxes will be in English 
language.  
For further details of IMP labelling, please refer to the study specific IMP-Manual. 
9.3 
Dose Regimen 
All 48 eligible participants will receive at their first treatment MV-ZIKA-RSP, at Visit 1 (day 0), 
and 4 weeks later the second treatment either with MV-ZIKA-RSP or placebo, at Visit 2 (day 
28 ±3)  
 
Visit 
VS 
Visit 1 
Visit 2 
Visit 3 
Visit 4 
Study Day 
 
0 
28 ±3 
56 ±3 
182 ±14 
Study Treatment 
 
 
 
 
 
 
The 4 treatments groups (A, B, C and D) differ in the concentration of MV-ZIKA-RSP, as well 
as, in single or double shot treatment regimen. 
 
Group A: 
MV-ZIKA-RSP 1 x105 (0.5 log) TCID50 /dose 
day 0 and day 28  
 
Group B: 
MV-ZIKA-RSP 2,5 x104 (0.5 log) TCID50 /dose 
day 0 and day 28  
 
 
 
 
Group C:  
MV-ZIKA-RSP 1 x105 (0.5 log) TCID50 /dose 
day 0 and placebo day 28 
 
 
 
 
Group D: 
Placebo  
day 0 and day 28 
9.4 
Justification of Clinical Dose Regime 
Two different doses of MV-ZIKA: 2,5x104 (± 0.5 log) - low dose- and 1x105 (± 0.5 log) -high 
dose- 50% tissue culture infective dose (TCID50) will be assessed in healthy adults for safety 
and immunogenicity as determined by the induction of functional, neutralizing antibodies 
(Table 4). A dose of 2x106 TCID50 was assessed for toxicity in a GLP compliant repeated dose 
toxicity study. The 20-fold higher dose compared to the clinical dose showed no adverse local 
or systemic effects. 
The clinical dose was selected based on findings in non-clinical pharmacology studies, clinical 
dose finding studies of comparable vaccine candidates and on assessment for cost-effective 
manufacturing of a vaccine for use in low-and middle-income countries. A dose of 1x105 TCID50 
06CCVP
06G6F2

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH
LQ1+3FRQIHUUHGSURWHFWLRQDJDLQVW=LNDYLUXVFKDOOHQJH,QDGGLWLRQFOLQLFDOWULDORQD
FRPSDUDEOH PHDVOHV YHFWRUEDVHG YDFFLQH FDQGLGDWH DJDLQVW &KLNXQJXQ\D YLUXV &+,.9
ZDVHIIHFWLYHDQGVDIHLQWKHKHUHSURSRVHGGRVHUDQJH$GRVHRI[7&,'LQGXFHGXS
WRVHURFRQYHUVLRQLQKHDOWK\DGXOWVXEMHFWVDIWHUWZRLPPXQL]DWLRQV

+RZHYHUDUHGXFWLRQWRDSRWHQWLDOO\
KLJKO\ HIIHFWLYH ORZHU GRVH ZLOO VXEVWDQWLDOO\ DIIHFW PDQXIDFWXULQJ FRVWV =LND YLUXV LV
SUHGRPLQDQWO\HQGHPLFLQORZDQGPLGGOHLQFRPHFRXQWULHV7KXVDYDFFLQHZLOOQHHGWRPHHW
WKHTXDOLWDWLYHDQGHFRQRPLFDOUHTXLUHPHQWV



7DEOH'RVHUHJLPH
7UHDWPHQW
*URXS
1
9DFFLQH
)RUPXODWLRQ
9DFFLQHRQ
GD\
3ODFHER
RQGD\
$

09=,.$563[PO
OLTXLGIUR]HQ


%

09=,.$563[PO
OLTXLGIUR]HQ


&

09=,.$563[PO
OLTXLG
IUR]HQSODFHER
OLTXLG

3ODFHERRQ
GD\
'

3ODFHERPO
OLTXLG
3ODFHERRQ
GD\
3ODFHERRQ
GD\

6WRUDJHDQG7UDQVSRUW&RQGLWLRQV
7KH,03PXVWEHVWRUHGXQGHUFRQWLQXRXVO\WHPSHUDWXUHFRQWUROOHGFRQGLWLRQLQDORFNDEOH
URRPRUORFNDEOHIUHH]HUZLWKOLPLWHGDFFHVV
7KH PLQLPXP DQG PD[LPXP WHPSHUDWXUH ZLOO EH UHFRUGHG GDLO\ LQ WKH WHPSHUDWXUH ORJV
WKURXJKRXWWKHVWXG\H[FHSWRQZHHNHQGVDQGKROLGD\VLIPLQPD[WKHUPRPHWHUZLOOEHXVHG
,QFDVHWKHIUHH]HUIULGJHLVFRQQHFWHGWRDQDODUPV\VWHPDQGFRUUHVSRQGLQJFRSLHVRIWKH
UHDGRXW FDQ EH ILOHG D PLQPD[ WHPSHUDWXUH GRHV QRW QHHG WR EH UHFRUGHG GDLO\ ,W LV
DFFHSWDEOHWRXVHDVLWHVSHFLILFWHPSHUDWXUHORJZKLFKFRYHUVDOOWKHUHTXLUHGLQIRUPDWLRQ
09=,.$563KDYHWREHNHSWLQWKHRXWHUSDFNDJHWREHSURWHFWHGIURPOLJKW
7KHOLTXLGIUR]HQIRUPXODWLRQRI09=,.$563ZLOOEHVWRUHGDQGWUDQVSRUWHGDW&DQG
VKDOOEHDGPLQLVWHUHGZLWKLQPLQDIWHUSUHSDUDWLRQWKHUHDG\WRXVHIRUPXODWLRQFDQEH
VWRUHGDW&IRUXSWRKRXU
7KHVDOLQHVROXWLRQDQGVWHULOHZDWHUGLOXHQWZLOOEHVWRUHGDFFRUGLQJWRLQVWUXFWLRQOHDIOHW
,WLVXQGHUWKHVLWH¶VUHVSRQVLELOLW\WRHQVXUHWKHVWRUDJHFRQGLWLRQVDFFRUGLQJWRWKHVXPPDU\
SURGXFWFKDUDFWHULVWLFV
$IWHUDGPLQLVWUDWLRQDOOXVHGFRQWDLQHUVSURSHUO\ODEHOOHGZLWKVXEMHFW,'DQGDSSURSULDWHYLVLW
LGHQWLILHU HJ YLVLW QXPEHU GDWH RI DGPLQLVWUDWLRQ ZLOO EH VWRUHG DW VLWH XQWLO FKHFNHG IRU
DFFRXQWDELOLW\E\WKHXQEOLQGHGPRQLWRU
&&93
CCI
*)

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 51/82 
9.6 
IMP Preparation and Distribution 
MV-ZIKA-RSP and placebo will be prepared and administered by unblinded, authorised and 
qualified staff members (investigator, pharmacist, or nurse) otherwise not involved in the 
conduct of the trial after randomisation. Preparation of MV-ZIKA-RSP for injection will be 
performed in a dedicated lab appropriate for handling of GMOs applying biosafety level 2 
(BSL2) standards although MV-ZIKA-RSP are assessed as a biosafety level 1 product. BSL2 
is applied because there is a negative system biological safety level 2 cabinet available within 
a containment room, which usually is used for preparing IMPs. 
MV-ZIKA-RSP and placebo will be exclusively used for the present clinical trial and will only 
be administered to participants enrolled in the study.  
For detailed description of receipt, storage, preparation, accountability, and destruction of IMP, 
please refer to the study specific IMP-Manual. 
9.7 
GMO Handling 
MV-ZIKA-RSP vaccine are a genetically modified organism requiring biosafety level 1 
standards. Nevertheless, preparation and disposal of IMP will be performed in a dedicated lab 
according to applicable hygiene standards of biosafety level 2. Needles and syringes that have 
been in contact with MV-ZIKA-RSP, as well as all other potentially contaminated materials, will 
be collected in dedicated containers and will be destroyed in a safe manner.  
This study will be conducted under regulations for contained use of a GMO.  
9.8 
Route and Method of Administration of IMP 
All participants will receive intramuscular (i.m.) injections of MV-ZIKA-RSP vaccine or placebo 
in the deltoid region of the non-dominant arm (preferred) or the dominant arm. In case of 
ongoing local AEs from previous vaccinations at the respective injection site, vaccination in the 
contra-lateral arm should be performed. 
 
9.9 
Treatment Compliance 
The study medication will be prepared at site and administered during the study Visit 1 and 2. 
Each treatment will be documented in the subject’s charts, the eCRF, an IMP Preparation Log 
and an IMP Inventory Log. Participants must receive all assigned vaccinations to be 
considered compliant. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 52/82 
 
9.10 Study Drug Accountability 
IMP accountability will be performed throughout the entire study, starting with the initial receipt 
of medication. 
The authorised, unblinded study staff members will confirm the number and condition of 
received vials, by signing and dating an IMP receipt form.  
Upon receipt an IMP inventory log will be kept current by the site, detailing the batch numbers, 
dates and quantity of IMP obtained, used for administration on a per subject basis and 
destroyed or returned to the sponsor. This documentation will only be available to an unblinded 
monitor verifying drug accountability during the study.  
At the end of the study, all unused and used MV-ZIKA-RSP vials (marked with subject ID and 
visit number) will be returned to the sponsor or destroyed by the investigator in a confidential 
manner, after final reconciliation and confirmation of correctness of drug accountability. 
9.11 Prior and Concomitant Therapy 
9.11.1 
Permitted Prior and Concomitant Therapy 
Any prior vaccination within the last three years prior screening and any medication within 30 
days prior screening has to be documented.  
Any medication taken during the study up to Visit 4 (study day 182) has to be reported to the 
investigator and will be documented.  
Additionally, any treatment that will be considered necessary for the subject’s welfare may be 
given at the discretion of the Investigator. All concomitant medications must be reported in the 
appropriate section of the eCRF along with dosage information, dates of administration, and 
reasons for use. Generic names for concomitant medication should be preferred, if possible. 
All concomitant medications will be coded using the WHO Drug Dictionary. 
9.11.2 
Forbidden Prior and Concomitant Therapy 
Treatments with non-study licensed vaccines within 4 weeks prior to first study vaccination 
until the end of the study (day 182) are not allowed. 
Use of immunosuppressive drugs like corticosteroids (excluding topical preparations) within 
30 days prior to the first vaccination or anticipated use before completion of the study (day 
182) is not allowed. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 53/82 
Receipt of blood products or immunoglobulins within 120 days prior to the Screening Visit or 
anticipated receipt of any blood product or immunoglobulin before completion of the study (day 
182) is not allowed.  
Participants must be asked about concomitant medication and vaccinations at each visit. Any 
concomitant medication or vaccination must be documented. 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 54/82 
10. SAFETY REPORTING 
10.1 Definitions 
10.1.1 
Adverse Event (AE) 
Any untoward medical occurrence in a participant to whom an investigational medicinal product 
(IMP) has been administered, not necessarily caused by or related to that product. 
An AE can therefore be any unfavourable or unintended sign, abnormal laboratory finding, 
symptom or disease temporally associated with the use of an IMP whether or not considered 
related to the IMP. 
10.1.2 
Clinical Laboratory Adverse Event 
A clinical laboratory AE is any clinical laboratory abnormality that suggests a disease and/or 
organ toxicity and is of a severity, which requires active management (i.e., changes of dose, 
discontinuation of drug, more frequent follow-up or diagnostic investigation). 
10.1.3 
Adverse Reaction (AR) 
An untoward and unintended response in a participant to an investigational medicinal product 
which is at least possibly related to any dose administered to that participant. 
All cases judged by either the reporting medically qualified professional or the sponsor as 
having a reasonable suspected causal relationship to the trial medication qualify as adverse 
reactions. 
10.1.4 
Serious Adverse Event (SAE) 
A serious adverse event is any untoward medical occurrence that: 
• 
results in death 
• 
is life-threatening 
• 
requires inpatient hospitalisation or prolongation of existing hospitalisation 
• 
results in persistent or significant disability/incapacity 
• 
consists of a congenital anomaly or birth defect 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 55/82 
Other ‘important medical events’ may also be considered serious if they jeopardise the 
participant or require an intervention to prevent one of the above consequences. 
The term "life-threatening" in the definition of "serious" refers to an event in which the 
participant was 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 were more severe. 
Any pregnancy occurring during the clinical trial and the outcome of the pregnancy will be 
recorded and followed up for congenital abnormality or birth defect, at which point it would fall 
within the definition of “serious”. 
NOTE: to avoid confusion or misunderstanding of the difference between the terms “serious” 
and “severe”, the following note of clarification is provided: “Severe” is used to describe the 
intensity of a specific adverse event, while “seriousness” is used for defining regulatory 
reporting obligations as supplied above. 
10.1.5 
Suspected Unexpected Serious Adverse Reaction (SUSAR) 
A serious adverse reaction, the nature, severity, or outcome of which is not consistent with the 
applicable product information.  
10.1.6 
Causality 
The relationship of each AE to the IMP must be determined by a medically qualified person 
according to the following definitions: 
Definitely: Temporal relationship to the administration of the study drug and course following a 
known reaction pattern 
Probably: Good reasons and sufficient documentation to assume a causal relationship 
Possibly: A causal relationship is conceivable and cannot be dismissed 
Unlikely: The event is most likely related to an etiology other than the trial treatment 
Not Related: No temporal relationship to the administration of the drug or other factors have 
caused the event 
10.1.7 
Severity 
AEs must be graded by a medically qualified person as being mild, moderate, severe, or life-
threatening and their approximate duration given. Definitions of severity are as follows: 
Mild: an AE that requires minimal or no treatment and does not interfere with daily activities. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 56/82 
Moderate: an AE that is sufficiently discomforting to interfere with normal activities. 
Severe: an AE that is incapacitating or prevents normal activities and may require systemic 
drug therapy or other treatment. 
Potentially life threatening:  an AE that requires immediate intervention to prevent death.  
"life-threatening" refers to an AE in which the subject was at risk of death at the time of the 
event, it does not refer to an AE which hypothetically might have caused death if it were more 
severe.  
Please see Appendix 1 for stencil to be used for grading local injection site reactions and Table 
5 and 6 for grading of systemic and local reactions. 
 
10.1.8 
Expectedness 
Expectedness will be determined considering the current Investigators’ Brochure. 
Expected: an AE that is listed in the current Investigator's Brochure of MV-CHIK. 
Unexpected: an AE that is not listed in the current Investigator's Brochure or differs due to 
greater severity or greater specificity. 
All SAEs assessed as unexpected and suspected to be related to the IMP qualify for a SUSAR 
(suspected unexpected serious adverse reaction) and require expedited reporting. 
10.1.9 
Outcome  
Recovered: A subject has recovered from an AE,when all signs or symptoms returned to 
normal. 
Stabilised: An AE is stabilised when, according to the investigator, the subject is in a clinically 
stable condition. This term should only be used for chronic conditions and for a given subject 
only when he/she has completed the study. 
Recovered with sequelae: As a result of the SAE, the subject is suffering from persistent or 
significant disability/incapacity (e.g. became blind, deaf, paralysed). Any AE recovered with 
sequelae should be rated as an SAE since an SAE criterion is fulfilled. 
Not Recovered: An AE currently ongoing. 
Ongoing at final examination: An AE ongoing at the subject’s last visit. 
Died: An AE that caused death. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 57/82 
 
10.2 Recording of Adverse Events 
All AEs occurring during the treatment period starting after the first vaccination at Visit 1 until 
termination of Visit 4 (day 182) observed by the investigator or reported by the subject (verbally 
or in the subject diary), will be recorded, whether or not related to the study treatment. 
AEs will be requested at each visit during the treatment period up to Visit 3 (day 56) and 
additionally solicited by a subject’s diary for 7 days after each vaccination. (see section Subject 
Diary) 
The participants will be asked generally (e.g. "How are you?") as well as specifically (e.g. “Have 
you experienced any problems since the last visit?”).  
The investigator must record all adverse events in the eCRF. One single adverse event page 
must be used per adverse event, from start to resolution, stating the following information:  
AE description, date of onset and end date, severity, relatedness, seriousness, and action 
taken. Please consider definitions of section 10.1. Follow-up information should be provided, 
as necessary.  
If possible, the investigator should record the diagnosis. If a diagnosis is not available, the 
investigator should record each sign and symptom as individual adverse events. However, as 
soon as a diagnosis becomes available, all signs and symptoms should be summarised and 
replaced by the underlying adverse event term (e.g. common cold instead of separate AEs for 
headache, rhinorrhoea, arthralgia, fever etc.) 
The AE should be documented stating the highest severity (i.e. if an AE goes from mild severity 
to moderate severity or vice versa, the whole episode of the AE should be reported with the 
moderate severity). Please consider Table 5 and 6 for grading local injection site reactions and 
systemic reactions. 
For causality reporting purposes, the categories "definitely", "probably" and "possibly" qualify 
an event as "suspected" adverse reaction. AEs with missing causality assessment will be 
regarded as possibly related unless further specified. 
Any medical condition that is present at screening will be considered as medical history and 
not reported as an AE. However, if the subject’s condition deteriorates at any time during the 
study, it will be recorded as an AE. 
It will be left to the investigator’s clinical judgment to decide whether an AE requires the 
subject’s removal from the study. In case of discontinuation, the subject will undergo an early 
termination visit. 
06CCVP
06G6F2

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH
 5HSRUWLQJRI6HULRXV$GYHUVH(YHQWV
7KHLQYHVWLJDWRUVKRXOGUHSRUWDOO6$(VRFFXUULQJGXULQJWKHWUHDWPHQWSHULRGZKHWKHURUQRW
UHODWHGWRWKH,03ZLWKLQKRXUVRIWKHVLWHVWXG\WHDPEHFRPLQJDZDUHRIWKHHYHQWWRWKH
$VVLJQ6DIHW\'HVNE\FRPSOHWLQJDQ6$(5HSRUW)RUP
7KHFRPSOHWHG6$(5HSRUW)RUPVKRXOGEHVXEPLWWHGWRWKH$VVLJQ6DIHW\'HVNHLWKHUE\ID[
RUDVSGISHUHPDLO
)D[
(PDLO

)RUXUJHQWTXHVWLRQVSOHDVHFDOOWKHKRXUV6DIHW\+RWOLQH
8QGHU FHUWDLQ FLUFXPVWDQFHV WKH ILUVW QRWLILFDWLRQ FDQ EH GRQH E\ SKRQH QHYHUWKHOHVV D
ZULWWHQ6HULRXV$GYHUVH(YHQW5HSRUW)RUPVKRXOGEHVXEPLWWHGIRUFRQILUPDWLRQWR$VVLJQ
6DIHW\'HVN
$IWHUWKHLQLWLDO6$(UHSRUWWKHLQYHVWLJDWRUZLOOIROORZXSSURDFWLYHO\HDFKVXEMHFWDQGSURYLGH
IXUWKHULQIRUPDWLRQOLNHFRSLHVRIKRVSLWDOFDVHUHSRUWVODERUDWRU\UHSRUWVRUDXWRSV\UHSRUWV
WRWKH$VVLJQ6DIHW\'HVNZLWKLQKDIWHUUHFHLSW
$VVLJQ6DIHW\'HVNZLOODVVHVVLQLWLDODQGIROORZXS6$(UHSRUWVIURPWKHVLWHIRUH[SHFWHGQHVV
ZKLFKZLOOEHGHWHUPLQHGEDVHGRQWKHPRVWUHFHQWHGLWLRQRIWKH,QYHVWLJDWRU¶V%URFKXUH
5HSRUWVRIVXVSHFWHGXQH[SHFWHGVHULRXVDGYHUVHUHDFWLRQV686$5VZLOOEHUHSRUWHGLQDQ
H[SHGLWHGPDQQHUWRWKH&$DVUHTXLUHGSHUORFDOUHJXODWLRQVZLWKLQWKHUHTXLUHGWLPHIUDPH
7KHRULJLQDOFRS\RIWKH6$(5HSRUW)RUPDQGWKHID[FRQILUPDWLRQVKHHWPXVWEHNHSWZLWK
WKHGRFXPHQWDWLRQDWWKHVWXG\VLWH(DFK6$(KDVDOVRWREHGRFXPHQWHGLQWKHH&5)DVD
VHULRXVDGYHUVHHYHQWLQWKHFRUUHVSRQGLQJ$(VHFWLRQ

 )ROORZXSRI$GYHUVH(YHQWV
'XULQJDQGIROORZLQJDVXEMHFW¶VSDUWLFLSDWLRQLQDFOLQLFDOWULDOWKHLQYHVWLJDWRUZLOOHQVXUHWKDW
DGHTXDWHPHGLFDOFDUHLVSURYLGHGWRWKHVXEMHFWIRUDQ\DGYHUVHHYHQWVLQFOXGLQJFOLQLFDOO\
VLJQLILFDQWODERUDWRU\YDOXHVUHODWHGWRWKHWULDO
$OOQRQVHULRXVDQGVHULRXVDGYHUVHHYHQWVZLOOEHIROORZHGXQWLOWKHVXEMHFWKDVUHFRYHUHG
VWDELOLVHGUHFRYHUHGZLWKVHTXHODHRUGLHGXQWLOHQGRIWKHVWXG\
)ROORZXSLQIRUPDWLRQDERXWDSUHYLRXVO\UHSRUWHG6$(PXVWDOVREHUHSRUWHGZLWKLQKRXUV
RIWKHLQYHVWLJDWRUUHFHLYLQJLW7KHLQLWLDO6$(UHSRUWIRUPVKRXOGEHXVHGVWDWLQJWKDWWKLVLVD
&&93
PPD
PPD
*)

09=,.$563


(XGUD&73URWRFRO
9HUVLRQ6HSWHPEHU





3DJH
IROORZXSUHSRUWWRWKHSUHYLRXVO\UHSRUWHG6$(DQGJLYLQJWKHQXPEHUDQGGDWHRIWKHIROORZ
XS UHSRUW 7KH LQIRUPDWLRQ SURYLGHG VKRXOG GHVFULEH ZKHWKHU WKH HYHQW KDV UHVROYHG RU
FRQWLQXHVLIDQGKRZLWZDVWUHDWHGDQGZKHWKHUWKHVXEMHFWFRQWLQXHGRUGLVFRQWLQXHGVWXG\
SDUWLFLSDWLRQLIQRWDOUHDG\VWDWHGLQWKHLQLWLDOUHSRUW 
 
 5HSRUWLQJRI686$5V
$ VHULRXV DGYHUVH UHDFWLRQ WKDW LV QRW H[SHFWHG DFFRUGLQJ WR WKH FXUUHQW ,% DQG ZHUH WKH
FDXVDOLW\ LV DVVHVVHG DV GHILQLWHO\ SUREDEO\ RU SRVVLEO\ TXDOLILHV DV VXVSHFWHG
XQH[SHFWHGVHULRXVDGYHUVHUHDFWLRQ
2QEHKDOIRIWKHVSRQVRU$VVLJQ6DIHW\'HVNZLOOEHUHVSRQVLEOHWRIXOILOWKHUHTXLUHPHQWVRI
H[SHGLWHGUHSRUWLQJRIDOO686$5VVXVSHFWHGXQH[SHFWHGVHULRXVDGYHUVHUHDFWLRQVWRWKH
&RPSHWHQW$XWKRULWLHV(WKLFV&RPPLWWHHVDQGSULQFLSDOLQYHVWLJDWRUVFRQFHUQHGDFFRUGLQJWR
QDWLRQDODQG(XURSHDQODZDQGUHJXODWLRQV
)RUIDWDODQGOLIHWKUHDWHQLQJ686$5VWKLVZLOOEHGRQHQRODWHUWKDQFDOHQGDUGD\VDIWHUWKH
VSRQVRURUGHOHJDWHLVILUVWDZDUHRIWKHUHDFWLRQ$Q\DGGLWLRQDOUHOHYDQWLQIRUPDWLRQWRWKH
LQLWLDOUHSRUWWKDWZDVQRWDYDLODEOHZLWKLQWKHFDOHQGDUGD\VZLOOEHUHSRUWHGZLWKLQDGGLWLRQDO
FDOHQGDUGD\V$OORWKHU686$5VZLOOEHUHSRUWHGZLWKLQFDOHQGDUGD\V
7UHDWPHQWFRGHVZLOOEHXQEOLQGHGIRUVSHFLILFSDUWLFLSDQWV
3ULQFLSDOLQYHVWLJDWRUVZLOOEHLQIRUPHGRIDOO686$5VUHODWHGWRWKHUHOHYDQW,03IRUDOOVWXGLHV
FRQGXFWHGZLWKWKHVDPH,03ZKHWKHURUQRWWKHHYHQWRFFXUUHGLQWKHFXUUHQWWULDO

 5HSRUWLQJRI3UHJQDQFLHV
:RPHQVKRXOGQRWEHFRPHSUHJQDQWGXULQJWKHWUHDWPHQWSHULRGRIWKHVWXG\,IDVXEMHFW
EHFRPHVSUHJQDQWZLWKLQWKHWUHDWPHQWSHULRGXSWRGD\VKHVKRXOGLPPHGLDWHO\LQIRUP
WKHLQYHVWLJDWRUDERXWWKHSUHJQDQF\
7KHLQYHVWLJDWRUZLOOFRPSOHWHDFRUUHVSRQGLQJ3UHJQDQF\5HSRUW)RUPDQGVHQGWKHIRUP
ZLWKLQKRXUVDIWHUEHFRPLQJDZDUHWR$VVLJQ6DIHW\'HVNYLDID[RUDVSGISHUHPDLO
)D[
(PDLO

7KHILUVWQRWLILFDWLRQFDQEHGRQHYLDSKRQHKRXUV6DIHW\+RWOLQH
&&93
PPD
PPD
*)

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 60/82 
Nevertheless, a written Pregnancy Report Form must be submitted for confirmation. The 
subject should attend follow-up visits as planned. In addition, participants who become 
pregnant within the treatment period up to day 56 (and who received at least one dose) will be 
followed until the pregnancy outcome, even if this occurs after the study is completed. 
Pregnancy itself is not regarded as an AE unless there is a suspicion that the IMP may have 
interfered with the effectiveness of a contraceptive medication. However, the outcome of all 
pregnancies that begin before day 56 (spontaneous miscarriage, elective termination, normal 
birth or congenital abnormality) will be followed up and documented even if the subject was 
discontinued from the study. 
A pregnancy outcome of a congenital abnormality/birth defect would qualify as an SAE and 
will be reported to Assign Safety Desk for processing to the regulatory authority. A pregnancy 
outcome of an elective abortion without evidence of complications would not be processed as 
an AE. 
 
10.7 Development Safety Update Reports 
Assign Data Management and Biostatistics GmbH will submit a DSUR once a year throughout 
the clinical trial to the competent authority (AGES), ethics committee, according to applicable 
regulations and requirements. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 61/82 
11. ASSESSMENT OF SAFETY  
Adverse event rates compared between 4 different treatment groups will be used to evaluate 
the secondary study objective. Please also refer to Table 2, summarising study procedures 
including safety assessments.  
11.1 Safety Laboratory Parameter Assessments 
Haematology, blood chemistry, coagulation parameters and urinalysis will be assessed for 
subject’s safety at screening, Visit 2, Visit 4 and additionally at Early Termination (if applicable). 
Please refer to section 10. 
11.2 Symptom-directed Physical Examination  
A full physical exam will be performed at screening and a system based physical examination 
will be done, if necessary, according to findings in a symptom-directed physical examination 
at Visit 1,2 and 3. This means if a symptom is reported by the subject, a system-based 
assessment will be performed, if needed, for a detailed check of the affected body system.  
Any symptom reported after first vaccination, including worsening of pre-existing conditions 
(i.e. medical history/concomitant diseases), will be recorded as AE. 
11.3 System-based Physical Examination 
All participants will undergo a system-based physical examination, including but not limited to 
assessment of general appearance and skin, head/eyes/ears/nose/throat, respiratory system, 
cardiovascular system, abdominal and gastrointestinal system, musculoskeletal system, 
neurological system and lymph nodes. 
This system-based physical examination will only be performed if necessary, according to 
findings of the symptom-directed physical exam at Visit screening as well as Visits 1,2 and 3. 
11.4 Vital Signs and Body Temperature 
Systolic and diastolic blood pressure and pulse rate as well as body temperature will be 
recorded at screening and Visits 1,2 and 3 with the subject at rest in a sitting position. 
11.5 Post Vaccination Reactogenicity Assessments 
All participants enrolled in the study and who receive at least one study treatment (MV-ZIKA-
RSP or placebo) will be followed up 28 days after each study treatment. Solicited and 
unsolicited AEs will be recorded at each visit until the end of the study at Visit 4. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 62/82 
11.6 Local and Systemic Tolerability 
The local tolerability (i.e. injection site reactions) will be inspected and evaluated by the 
investigator one hour after each IMP administration.  
Additionally, local tolerability (i.e. inspection of the previous injection site) will be assessed at 
the following visits and prior to application of the subsequent vaccination at Visit 2 (day 28). 
Furthermore, local tolerability will be evaluated by the participants in the subject diaries for 7 
days starting 6 hours after each treatment.  
The grading of local reaction will be performed by means of a stencil (provided to the subject 
as shown in Appendix 1) and per "Guidance for Industry: Toxicity Grading Scale for Healthy 
Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical Trials", modified, 
reflecting the guidance of the Brighton Collaboration (Table 5). 
Findings in the local tolerability examination by the investigator as well as local tolerability 
findings recorded in the subject diary will be covered as adverse events in the eCRF.  
 
11.7 Subject Diary 
The participants will be instructed to complete a diary for 7 days after each vaccination to 
document local and systemic tolerability as well as body temperature. 
Solicited local and solicited systemic AEs will be assessed by the participants themselves by 
checking for presence of the listed symptoms and measuring the size of the affected area 
where appropriate (a template will be provided for local reaction grading, as shown in Appendix 
1). The diary will also provide space for recording unsolicited AEs and concomitant medication.  
Recording should be done approximately at the same time each day, starting on the day of 
vaccination. The first entry should be made 6 hours 1h after vaccination at Visit 1 and Visit 2.  
The participants’ diaries will be collected and verified for completeness by the investigator at 
Visit 2 and Visit 3. Any AE recorded in the subject diaries will be entered in the eCRF by the 
investigator or authorised delegates. The investigator will re-evaluate the severity of the 
reported local and systemic AEs according to the below Tables 5 and 6. 
Diary solicited systemic AEs include nausea, vomiting, headache, and fatigue, also to be 
documented in the eCRF.  
In addition, body temperature will be recorded daily for 7 days and fever will be graded by the 
investigator according to the FDA Guidance for Industry. For details see below Table 6.Table 
5. Grading of local reactions 
Local reaction 
to injectable 
product 
Mild  
(grade 1) 
Moderate  
(grade 2) 
Severe  
(grade 3) 
Potentially life 
threatening 
(grade 4) 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 63/82 
Pain 
Does not 
interfere with 
activity 
Interfere with activity 
or repeated use of 
non-narcotic pain 
reliever 
Prevents daily activity 
or repeated use of non-
narcotic pain reliever 
Emergency room 
(ER) visit or 
hospitalisation 
Tenderness 
Mild pain to touch 
Pain with movement 
Significant pain at rest 
ER visit or 
hospitalisation 
Erythema/ 
Redness1 
≤ 5 cm 
5.1–10 cm 
> 10 cm 
Necrosis or exfoliate 
dermatitis 
Induration/ 
Swelling2 
≤ 5 cm and does 
not interfere with 
activity  
5.1–10 cm or 
interfere with activity 
> 10 cm or prevents 
daily activity 
Necrosis 
Itching 
Does not 
interfere with 
activity 
Interferes with activity 
or repeated use of 
non-narcotic pain 
reliever 
Prevents daily activity 
or repeated use of anti-
inflammation and pain-
relieving ointment 
Emergency room 
(ER) visit or 
hospitalisation 
1 In addition to grading the measured local reaction at the greatest single diameter, the measurement should be recorded as a 
 continuous variable. 
2  Swelling should be evaluated and graded using the functional scale as well as the actual measurement. 
 
Table 6. Grading of systemic reactions 
Systemic 
Reaction 
(General) 
Mild  
(grade 1) 
Moderate  
(grade 2) 
Severe  
(grade 3) 
Potentially life 
threatening 
(grade 4) 
Nausea/ 
vomiting 
No interference with 
activity or 1-2 
episodes/ 24 hours 
Some interference 
with activity or >2 
episodes/ 24 hours 
Prevents daily activity, 
requires outpatient IV 
hydration 
Emergency room 
(ER) visit or 
hospitalisation for 
hypotensive shock 
Diarrhoea 
2-3 loose stools or < 
400 gms/ 24hours 
4-5 stools or < 400-
800 gms/ 24hours 
6 or more watery 
stools or >800 gms/ 
24 hours or requires 
outpatient IV 
hydration 
ER visit or 
hospitalisation 
Headache 
No interference with 
activity 
Repeated use of 
non-narcotic pain 
reliever >24 hours or 
some interference 
with activity 
Significant; any use 
of narcotic pain 
reliever or prevents 
daily activity 
ER visit or 
hospitalisation 
Fatigue 
No interference with 
activity 
Some interference 
with activity 
Significant; prevents 
daily activity 
ER visit or 
hospitalisation 
Myalgia 
No interference with 
activity 
Some interference 
with activity 
Significant; prevents 
daily activity 
ER visit or 
hospitalisation 
Illness or 
clinical AE 
(as defined 
according to 
applicable 
regulations) 
No interference with 
activity 
Some interference 
with activity not 
requiring medical 
intervention 
Prevents daily 
activity and requires 
medical intervention 
ER visit or 
hospitalisation 
Fever (°C) 
38.0 - 38.4 
38.5 - 38.9 
39.0 - 40.0 
>40.0 
 
 
 
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 64/82 
12. STATISTICS 
The data will be analysed by Assign Data Management and Biostatistics GmbH. A Statistical 
Analysis Plan (SAP) will be finalised before database snapshot for the preliminary analysis 
providing a detailed description on the statistical methods and evaluation of study results. 
A preliminary analysis will be performed after all participants completed the treatment period 
and the final analysis will be conducted after the last subject completed the Post Treatment 
Safety Follow-Up. 
 
12.1 Preliminary Analysis 
A preliminary analysis including safety data will be performed after all participants completed 
Visit 3 (day 56) and therefore terminated the treatment period of the study. This analysis will 
cover the primary safety endpoint (adverse events up to day 56. The purpose of this analysis 
is to provide highly valuable data for further zika vaccine development. 
Results from this preliminary analysis will not have any impact on the study design or the 
participants during the follow up period of the study.  
12.2 Methods of Statistical Immunogenicity Analysis 
The immunogenicity analysis will compare the anti-zika-rsp VNT antibody geometric mean titre 
(GMT) in the Per Protocol (PP) analysis population between the treatment groups. GMTs and 
GMT ratios will be estimated by applying an analysis of variance including the factor treatment 
group. This will be done using log10 transformed data and taking the anti-log of the resulting 
point estimates for the least squares means, least squares means differences and the 
corresponding 2-sided 95% CIs.  
P-values will also be provided to compare GMTs between the treatment groups adjusted for 
multiple comparisons according to Tukey-Kramer. Seroconversion rates will be compared 
between groups using Fisher’s exact test.  
Seroconversion will be defined as anti-zika-rsp VNT titres ≥20. 
ZIKA-RSP- and measles- ELISA titres will be analysed as described above.  
12.3 Methods of Statistical Safety Analysis 
The analysis of safety will be performed in the Safety Population.  
AEs and concomitant diseases will be coded using the MedDRA coding dictionary.  
Solicited AEs (reactogenicity):  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 65/82 
Solicited local and systemic AEs gathered by the participants’ diaries will be summarised for 
the 7-day period, by AE grading (mild, moderate, severe) and by treatment group. The number 
and percentage of participants with AEs will be presented. Fisher exact tests will be provided 
for a comparison of AE rates between treatment groups in summary tables. 
Unsolicited AEs:  
Separately, the number and percentage of participants with unsolicited/spontaneous adverse 
events (AEs)and serious adverse events (SAEs) will be presented for each treatment group 
overall, and by system organ class/preferred term, by AE grade, and relatedness. Fisher exact 
tests will be provided for a comparison of AE rates between treatment groups. 
Laboratory values and vital signs as well as changes in laboratory values from baseline will be 
analysed descriptively by time point and treatment group.  
 
12.4  Determination and Justification of Sample Size 
A formal sample size calculation was not conducted. The sample size of 48 participants has 
been determined based on prior experience in evaluating the safety and immunogenicity of 
vaccines and is typical for early phase clinical studies. Sample size for this study was 
determined on grounds of feasibility and common practice in similar trials. 
12.5  Analysis Populations 
12.5.1 
Safety Population 
All safety analyses will be based on the safety population, defined as participants who entered 
into the study and received at least one IMP administration. All analysis based on the Safety 
Population will be carried out using the actual treatment received.  
12.5.2 
Modified Intent-to-Treat (mITT) Population 
The secondary immunogenicity analyses will be based on the modified ITT population. The 
modified intent-to-treat (mITT) analysis population is defined to include all participants 
randomised who receive at least one IMP administration. Participants will be analysed 
according to the treatment group they had been allocated to, rather than by the actual 
treatment they received. 
12.5.3 
Per-Protocol (PP) Population 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 66/82 
The immunogenicity analysis will be assessed primarily on the per-protocol (PP) population. 
The PP population includes participants without protocol deviations that could impact immune 
response. Examples that may lead to exclusion from the PP population are provided here 
(further criteria may be defined in the SAP): 
- Immunosuppressive drugs: Use of corticosteroids (excluding topical preparations) or 
immunosuppressive drugs within 30 days prior to first IMP administration, or anticipated 
use during the trial. 
- Participants with any confirmed immunosuppressive or immunodeficient condition, 
including human immunodeficiency virus (HIV), hepatitis A, B or C infection or a family 
history of congenital or hereditary immunodeficiency 
- Participants who received the wrong or no IMP 
These criteria for potential protocol violation are identified at the time of planning the study. 
However, during the course of the trial unforeseen events may occur or new scientific 
knowledge may become available, therefore final decisions on whether any protocol violation 
could impact immune response and thus lead to exclusion from the PP population will be made 
by the sponsor on a case by case basis in a blinded manner (and prior to study unblinding at 
the preliminary analysis). Sample testing issues may also lead to exclusion from the PP 
population for particular time points.  
 
12.6 Statistical Analysis Plan 
A Statistical Analysis Plan (SAP) will be written and finalised prior to the preliminary analysis. 
The SAP will provide a detailed description of the statistical methods and expand on the details 
provided in the protocol. Additional analyses may be added.  
 
12.7  Missing Data 
All attempts will be made to prevent missing data and generally, missing data will not be 
imputed, and analysis will be limited to observed values. However, for missing data in AE 
evaluation (e.g. missing information about severity or causality) a worst-case approach will be 
applied.  
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 67/82 
13. QUALITY CONTROL AND QUALITY ASSURANCE 
The trial will be conducted in accordance with the current approved protocol, ICH GCP 
guidelines, relevant regulations and standard operating procedures. 
 
13.1 Periodic Monitoring 
A designated Clinical Research Associate (CRA) will monitor study progress by scheduling 
and performing on-site study visits throughout the study including site initiation visit, several 
site monitoring visits and site close out visit. At regular intervals throughout the study 
monitoring visits will be performed to verify completeness, accuracy and consistency of data 
in the eCRFs, protocol adherence, adherence to GCP and the applicable regulatory 
requirements. Frequency and scope of source data verification will be determined before study 
start and detailed in the Monitoring Plan. 
Therefore, the monitor must have access to all source records needed to verify the entries on 
the eCRFs. The investigator will cooperate with the monitor to ensure that any discrepancies 
identified are resolved. 
The periodic monitoring will be performed by Neox Clinical Research. 
 
13.2 Audit and Inspection 
Upon request, the investigator will make all study-related source data and records available to 
a qualified quality assurance auditor mandated by the sponsor or to regulatory inspectors.  
The investigator or designee should contact the sponsor/CRO immediately upon 
announcement of an audit by the regulatory authority. He/she further agrees to fully cooperate 
with competent authorities and participate with audits conducted at a convenient time in a 
reasonable manner. 
The main purposes of an audit or inspection are to confirm that the rights and welfare of the 
participants have been adequately protected, to assess whether ethics, regulatory and quality 
requirements are met and to verify that all data relevant for the assessment of safety and 
immunogenicity of the investigational product have appropriately been reported to the sponsor. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 68/82 
14. ETHICS 
14.1 Ethical Conduct of the Study 
This study will be conducted in compliance with relevant regulatory requirements, ICH GCP 
guidelines of the Declaration of Helsinki for biomedical research involving human participants 
and the EU directive 2005/28/EC. 
 
14.2 Ethics Committees (EC) (AMG §§ 40,41) 
Prior to initiation of the study, the protocol, subject informed consent form, any other written 
information to be provided to the subject, any proposed advertising material, investigator’s 
brochure (IB), information about payments and compensation available to participants if not 
mentioned in the subject information, investigator’s current CV and other documentation 
evidencing qualifications, and other documents as required by the ethics committee should be 
submitted.  
Written approval/favourable opinion must be obtained from the IEC prior to commencement of 
the study. This approval should include a statement that these documents comply with GCP 
requirements and must identify the documents and versions reviewed. 
During the trial, the investigator must promptly report the following to the IEC: amendments to 
the protocol, updates to IB, unexpected SAEs where a causal relationship cannot be ruled out, 
notes of administrative changes, deviations to the protocol implemented to eliminate 
immediate hazards to the trial participants, new information that may affect adversely the safety 
of the participants or the conduct of the trial, annually written summaries of the trial status, and 
other documents as required by the local IEC. 
Amendments must not be implemented before approval/favourable opinion, unless necessary 
to eliminate immediate hazards to the participants. 
 
14.3 Competent Authority (CA) 
The CA will receive the protocol, amendments, reports on SAEs, and related relevant safety 
information, including the final report according to EU Directive and national regulations. 
Written approval/favourable opinion must be obtained from the CA prior to commencement of 
the study and for all substantial amendments to the original documents before implementation. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 69/82 
15. DATA HANDLING AND RECORD KEEPING 
15.1 Source Data and Records 
Source documents are where data are first recorded and from which participants’ eCRF data 
are obtained. These include, but are not limited to, original charts (participants’ records), 
laboratory and pharmacy printouts, diaries, and data from automated instruments. On all trial-
specific documents, other than the signed consent, the participant will be referred to by subject 
number, not by name. 
At least the following data will be documented in the source records: 
- Study identification, date of subject’s study entry and termination  
- Subject number 
- Documentation of informed consent procedure 
- Date of each study visit  
- Medical history, demographic data 
- Any examination findings, including local injection site reactions 
- Adverse events 
- Concomitant medication intake 
- Early withdrawal date and withdrawal reason, if applicable 
- Treatment dates  
- Completed subject diaries 
 
Source data entries must be made in accordance with GCP and local requirements.  
The investigator will permit study-related monitoring, audits, IRB/IEC review and regulatory 
inspections, by providing direct access to source data. Source records should be preserved 
for the maximum period of time required by local regulations. 
15.2 Periodic Monitoring 
A designated monitor will inspect the eCRFs at regular intervals throughout the study to verify 
completeness, accuracy and consistency of the data, protocol adherence, and adherence to 
Good Clinical Practice guidelines. The monitor should have access to all source records 
needed to verify the entries on the eCRFs. The investigator will cooperate with the monitor to 
ensure that any discrepancies identified are resolved 
15.3 Access to Data 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 70/82 
Direct access to source data and documents will be granted to authorised representatives from 
the sponsor, host institution and the competent authorities to permit trial-related monitoring, 
IRB/IEC review, audits and inspections. 
 
15.4 Data Collection 
During each study visit, the investigator will collect and maintain notes in the subject’s study 
records to document all procedures, significant observations and assessments, which are 
regarded as source data. Additionally, diaries and laboratory result reports signed and dated 
by the investigator, have to be kept within the subject’s records. Changes to information in the 
study record and other source documents will be initiated and dated on the day the change is 
made. All documents will be stored safely under confidential conditions. 
15.5 Electronic Case Report Forms (eCRFs) 
15.4.1 
eCRF Entries 
Information from the participants’ study records and other source documents will be entered 
into a 21 CFR Part 11-compliant validated electronic Case Report Form (eCRF) as soon as 
possible after each visit. The participants will be identified by a unique trial specific subject 
number. The name or any other identifying detail will NOT be included in the eCRF. 
Entries and corrections in the eCRF may only be performed by the investigator or authorised 
study site staff via personal user account and password for access. This password must be 
kept confidential and must only be used by the person to whom it was assigned to ensure that 
each entry/change can be allocated to the person who performed the entry/change. 
An automatic audit trail will log each data entry/change performed in the eCRF. 
The eCRF system includes internal quality checks, to identify data that appear inconsistent, 
incomplete or inaccurate. Maintenance of the eCRF and the study database will be performed 
by the data management centre at Assign Data Management and Biostatistics GmbH. 
15.4.2 
Changes to eCRF Data 
Necessary data changes of the eCRF data may be identified as follows: 
• 
Entries are checked by the eCRF during data entry or when the eCRF page has been 
submitted/saved. If the data does not fulfil particular quality criteria, a message will 
specify the type of problem and assist in its correction. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 71/82 
• 
Monitors may ask for correction of data during monitoring (e.g., if the eCRF entry does 
not match the source data). 
• 
Computerised data-check programs and/or manual checks will identify clinical data 
discrepancies. Corresponding queries aiming at the resolution of these discrepancies 
will be created within the eCRF system and the study site will be informed about new 
issues to be resolved on-line. 
All discrepancies will be resolved on-line directly by the investigator or by authorised staff. 
As long as an eCRF page is not locked, required data changes can be conducted by the 
investigator or authorised site staff at any time. 
15.4.3 
eCRF Entry Validation 
The principal investigator or the authorised delegate will thoroughly review the eCRF data and 
will finally certify the contents of the eCRF by electronically signing the eCRFs within the data 
capturing system directly. If a correction was made to the eCRF data after the investigator’s 
approval, the certification must be repeated after the changes were performed. 
15.5 Confidentiality of Subject’s Data 
The investigator will ensure that all subject’s information obtained during the conduct of the 
study will be kept confidential maintaining subject’s anonymity. 
On eCRFs or any other documents submitted to the sponsor, participants will only be identified 
by subject number and therefore remain confidential in all reports or publications related to the 
study. Documents revealing subject’s identification, e.g. subject identification log and original 
informed consent forms, will be maintained by the investigator in strict confidence, except to 
the extent necessary to allow checking by CRAs, the Sponsor or competent authorities. 
 
15.6 Record Maintenance, Archiving and Destruction 
The investigator is responsible for compiling and keeping all essential documents in the 
Investigator Site File (ISF) during the study and for the maximum period of time in accordance 
with ICH GCP guidelines and local regulatory requirements.  
The ISF has to be stored in a safe and secure location with restricted access to staff personnel 
only and will include all essential documents like the clinical protocol, competent authorities’ 
approvals, original informed consent forms, source data, IMP dispensing and accountability 
logs, participants’ logs and all correspondence pertaining to the study.  
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 72/82 
The investigator will agree to archive the study documentation and will not dispose of any 
records relevant to this study without prior written permission from the sponsor. If an 
investigator moves, withdraws, or retires, the responsibility for maintaining the records may be 
transferred to another person upon sponsor’s agreement. 
The sponsor or their representative will notify the investigator when study documents need no 
longer be retained. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 73/82 
16. CHANGES IN THE CONDUCT OF THE STUDY 
16.1 Protocol Amendments 
Any proposed changes to the protocol must be covered in a written amendment to the protocol 
and submitted to the appropriate ethics committees or competent authorities. Amendments 
may only be implemented after approval of the appropriate ethics committees or competent 
authorities have been obtained except where necessary to eliminate apparent immediate 
hazard to participants. 
 
16.2 Study Termination 
If the sponsor or the investigator decides to terminate the study before it is completed, they will 
notify each other in writing stating the reasons for early termination. In terminating the study, 
the sponsor and the investigator will ensure that adequate consideration is given to the 
protection of the participants’ interest. The investigator, sponsor or CRO will notify the relevant 
ethics committee and regulatory authority in writing in accordance with local requirements. 
Documentation will be submitted for filing in Central File and Investigator Site File. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 74/82 
17. DEVIATIONS FROM THE PROTOCOL  
17.1 Relevant Protocol Deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, ICH GCP, manuals or 
local requirements. The noncompliance may be either on the part of the subject, the 
Investigator, or the study site staff. Consequently, corrective actions are to be developed and 
implemented promptly.  
Important protocol deviations are deviations that might significantly affect the completeness, 
accuracy and/or reliability of the study data or that might significantly affect a subject’s rights, 
safety, or well-being. 
All protocol deviations will be listed in the study report and assessed as to their influence on 
the quality of the study analysis. No deviations from the protocol of any type will be made 
without complying with all the ethics committee’s or regulatory authority’s established 
procedures in accordance with applicable regulations. 
 
17.2 Serious Breaches 
The Medicines for Human Use (Clinical Trials) Regulations contain a requirement for the 
notification of "serious breaches" to the AGES within 7 days of the sponsor becoming aware 
of the breach. 
A serious breach is defined as “A breach of GCP or the trial protocol which is likely to affect to 
a significant degree:  
- the safety or physical or mental integrity of the participants of the trial, or 
- the scientific value of the trial”. 
If a serious breach is suspected the sponsor must be contacted within 1 working day. In 
collaboration with the principal investigator, the serious breach will be reviewed by the sponsor 
and, if appropriate, the sponsor will report it to the Regulatory Authority within 7 calendar days. 
 
17.3 Premature Subject Withdrawal 
Participants have the right to withdraw from the study at any time for any reason, without the 
need to justify. The investigator also has the right to withdraw participants in case of AEs, 
protocol violations or administrative reasons. Please refer to section 8.9 for details. 
Since an excessive rate of withdrawal can render the study inconclusively, the unnecessary 
withdrawal of participants must be avoided. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 75/82 
18. FINANCE AND INSURANCE 
18.1 Contractual and Financial Details 
The investigator, the sponsor and CRO will sign a clinical study agreement prior to the start of 
the study outlining overall sponsor and investigator responsibilities in relation to the study. The 
contract will describe costs for pharmacy, laboratory and other protocol-required services.  
Financial Disclosure Statements will be completed, as required by 21 CFR part 54. 
 
18.2 Insurance (AMG §32(2)) 
All participants participating in this clinical study will be insured through Themis Biosciences 
GmbH.  
 
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 76/82 
19. REPORTING AND PUBLICATION 
19.1 Clinical Study Report 
After all participants terminated their study participation (either by completion of Visit 4 (day 
182) or prematurely by early termination or are confirmed as lost to follow up), a clinical study 
report will be prepared by the sponsor or delegate in accordance with relevant guidelines. 
 
19.1 Publication Policy 
All results generated in this study will be considered to be strictly confidential. The investigators 
may not submit the results for publication or presentation without prior written permission of 
the sponsor. Authorship for any publication will be determined in mutual agreement. 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 77/82 
20. REVISION INDEX 
Section 
Modification 
Reason 
Version 1.7 (22 Apr 2019) 
N/A 
First version 
New Document 
Version 1.8 (13 Jun 2019) 
 
The Clinical Study Protocol was amended to comply with the 
requirements and feedback received from the Ethics 
Committee. Following changes have been performed:  
8.2 
Statistics: Request from the 
Ethic Committee: For safety 
reasons, only 6 patients 
should be included for the 
time being, and if no serious 
AE occur, the study will be 
continued. However, due to 
the randomization, it cannot 
be ruled out that all or most 
of these 6 patients will be 
assigned to the placebo 
arm. This appears 
problematic, so make sure 
that enough patients from 
the highest dose group are 
included in these 6 patients. 
Request implemented in the 
Protocol Section 8.2. 
Randomisation 
8.11.2 
Blood volumes drawn for the 
Immunogenicity testing 
updated. 
To specify more preciously 
the volumes. 
8.11.3 
Pregnancy test specified. 
To clarify the sample for 
pregnancy test. 
9.4 
Request of an explicit 
justification for selecting for 
the clinical trial a lower dose 
than used for animal 
experiments.  
Justification was included in 
the protocol (Section 9.4) 
Version 1.9 (26 Jun 2019) 
8.2 
Specification of the 
adequate representation of 
the randomization was 
added. 
Request from Ethic 
Committee 
Version 2.0 (25 Mar 2020) 
11.7 
Grading of local reactions 
was updated. 
The update was done to be 
in accordance with definition 
in the referenced FDA 
Guidance for Industry 
“Toxicity Grading Scale for 
Healthy Adult and 
Adolescent Volunteers 
Enrolled in Preventive 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 78/82 
Vaccine Clinical Trials”, 
September 2007. 
12.1 
Preliminary Analysis of 
Immunogenicity data will not 
be performed after all 
participants completed Visit 
3 (day 56). 
 
The reason is that the labs 
don’t have capacity for those 
tests due to the current 
Covid-19 pandemic 
situation. 
17.1 
Paragraph removed from 
this Section since do not 
proceed here. 
Paragraph removed. 
Version 2.1 (28 Sep 2020) 
1. 
PRNT50 was replaced with 
VNT. 
A Plaque Reduction 
Neutralization Test method 
was replaced with another, 
highly similar serum sample 
test termed Virus 
Neutralization Test. 
4.5 
5.2 
6.1 
6.2.2 
6.5 
8.5.1 
8.5.3 
8.5.4 
8.11.2 
Description of usage of a 
Plaque reduction 
neutralization assay was 
replaced with description of 
usage of a Virus 
Neutralization Test. 
8.11.4 
PRNT50 was replaced with 
VNT. 
12.2 
12.2 
Seroconversion 
will 
be 
defined as anti-zika-rsp VNT 
titres ≥20 10. 
Neutralizing antibodies were 
quantified with a slightly 
different assay than 
originally planned. This VNT 
has a Lower Limit of 
Quantification (LLOQ) of 20, 
requiring us to adapt the 
protocol to accommodate 
the more stringent cut-off. 
 
 
21. REFERENCES 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 79/82 
CDC/PAHO. Preparedness and Response for Chikungunya virus introduction in the Americas. 
www.paho.org/hq/index.php?option=com_docman&task=doc_download&gid=16984&Itemid=
&lang=en 
Brandler S, Lucas-Hourani M, Moris A, Frenkiel MP, Combredet C, Février M, Bedouelle H, 
Schwartz O, Desprès P, Tangy F. Pediatric measles vaccine expressing a dengue antigen 
induces durable serotype-specific neutralizing antibodies to dengue virus. PLoS Negl Trop Dis. 
2007;1(3):e96. 
Brandler S, Marianneau P, Loth P, Lacôte S, Combredet C, Frenkiel MP, Desprès P, Contamin 
H, Tangy F. Measles vaccine expressing the secreted form of West Nile virus envelope 
glycoprotein induces protective immunity in squirrel monkeys, a new model of West Nile virus 
infection. J Infect Dis. 2012;206:212-9.  
Brandler S, Ruffie C, Najburg V, Frenkiel MP, Bedouelle H, Desprès P, Tangy F. Pediatric 
measles vaccine expressing a dengue tetravalent antigen elicits neutralizing antibodies 
against all four dengue viruses. Vaccine. 2010;28:6730-9.  
Brandler S, Ruffié C, Combredet C, Brault JB, Najburg V, Prevost MC, Habel A, Tauber E, 
Desprès P, Tangy F. A recombinant measles vaccine expressing chikungunya virus-like 
particles is strongly immunogenic and protects mice from lethal challenge with chikungunya 
virus. Vaccine. 2013;31(36):3718-25.  
Cao-Lormeau VM et al, Emerg Infect Dis 2013; 20: 1085–86.  
Cauchemez S et al, The Lancet 2016  
Combredet C, Labrousse V, Mollet L, Lorin C, Delebecque F, Hurtrel B, McClure H, Feinberg 
MB, Brahic M, Tangy F. A molecularly cloned Schwarz strain of measles virus vaccine induces 
strong immune responses in macaques and transgenic mice. J Virol. 2003;77(21):11546-54.  
Commission Directive 2005/28/EC laying  down  principles  and  detailed  guidelines  for  good  
clinical  practice  as  regards  investigational medicinal   products   for   human   use,   as   well   
as   the   requirements   for   authorisation   of   the manufacturing  or importation  of such 
products; see 
https://ec.europa.eu/health/files/eudralex/vol-1/dir_2005_28/dir_2005_28_en.pdf 
Committee for Human Medicinal Products (CHMP): Note for Guidance on the Clinical 
Evaluation 
of 
Vaccines 
(CHMP/VWP/164653/2005); 
see: 
http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC50
0003875.pdf 
Desprès P, Combredet C, Frenkiel MP, Lorin C, Brahic M, Tangy F. Live measles vaccine 
expressing the secreted form of the West Nile virus envelope glycoprotein protects against 
West Nile virus encephalitis. J Infect Dis. 2005;191(2):207-14.  
Dick GWA et al, Trans R Soc Trop Med Hyg 1952; 46: 509–20 
Duffy MR et al, N Engl J Med 2009; 360: 2536–43.  
ECDC. Rapid risk assessment. Zika virus epidemic in the Americas: potential association with 
microcephaly 
and 
Guillain-Barré 
syndrome. 
Dec 
10, 
2015. 
http://ecdc.europa.eu/en/publications/ 
Publications/zika-virus-americas-association-with-
microcephaly- rapid-risk-assessment.pdf (accessed Feb 5, 2016).  
Enserik M (2008) Entomology. A mosquito goes global. Science 320: 864-866 
European Centre for Disease Prevention and Control (ECDC), December 10, 2015 
(http://ecdc.europa.eu/en/publications/Publications/zika-virus-americas-association-with-
microcephaly-rapid-risk-assessment.pdf) 
FDA Guidance for Industry: Toxicity Grading Scale for Healthy Adult and Adolescent 
Volunteers 
Enrolled 
in 
Preventive 
Vaccine 
Clinical 
Trials 
(Sept. 
2007) 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 80/82 
http://www.fda.gov/BiologicsBloodVaccines/GuidanceComplianceRegulatoryInformation/Guid
ances/Vaccines/ucm074775.htm 
Gubler D et al. Flaviviruses. In: Knipe DM, Howley PM, Griffin DE, Lamb RA, Martin MA, et al, 
eds. Fields Virology, 5th edn. Philadelphia, PA: Lippincott Williams & Wilkins Publishers, 
2007: 1155–227. 
Gerardin P, Barau G, Michault A, Bintner M, Randrianaivo H, et al. (2008) Multidisciplinary 
prospective study of mother-to-child chikungunya virus infections on the island of La Reunion. 
PLoS Med 5: e60. doi:10.1371/journal.pmed.0050060 
International Conference on Harmonisation of Technical Requirements for Registration of 
Pharmaceuticals for Human Use: ICH Harmonised Tripartite Guideline: Guideline for Good 
Clinical 
Practice 
E6 
(R1); https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products 
/Guidelines /Efficacy/E6/E6_R1_Guideline.pdf 
Jain M, Rai S, Chakravarti A (2008) Chikungunya: a review. Trop Doct 38: 70-72 
Lorin C, Mollet L, Delebecque F, Combredet C, Hurtrel B, Charneau P, Brahic M, Tangy F. A 
single injection of recombinant measles virus vaccines expressing human immunodeficiency 
virus (HIV) type 1 clade B envelope glycoproteins induces neutralizing antibodies and cellular 
immune responses to HIV. J Virol. 2004;78(1):146-57.  
Macnamara FN, Trans R Soc Trop Med Hyg 1954; 48: 139–45 
Msaouel P, Opyrchal M, Domingo Musibay E, Galantis E. Expert Oncolytic measles virus 
strains as novel anticancer agents Opin Biol Ther. 2013 Apr;13(4):483-502. 
Paul BJ, Pannarkady G, Moni SP, Thachil EJ. Clinical profile and long-term sequelae of 
Chikungunya fever. Indian Journal of Rheumatology. Volume 6, Issue 1, March 2011, 12–19. 
Ramsauer K, Schwameis M, Firbas C, Müllner M, Putnak RJ, Thomas SJ, Desprès P, Tauber 
E, Jilma B, Tangy F. Immunogenicity, safety, and tolerability of a recombinant measles-virus-
based chikungunya vaccine: a randomised, double-blind, placebo-controlled, active-
comparator, first-in-man trial. Lancet Infect Dis. 2015 May;15(5):519-27.  
Immunogenicity, safety, and tolerability of the measles-vectored chikungunya virus vaccine 
MV-CHIK: a double-blind, randomised, placebo-controlled and active-controlled phase 2 trial  
EC Reisinger, R Tschismarov, E Beubler, U Wiedermann, C Firbas, M Löbermann, A Pfeiffer, 
M Muellner, E Tauber, K Ramsauer. Immunogenicity, safety, and tolerability of the measles-
vectored chikungunya virus vaccine MV-CHIK: a double-blind, randomised, placebo-controlled 
and active-controlled phase 2 trial. Published online www.thelancet.com, November 5, 2018; 
http://dx.doi.org/10.1016/S0140-6736(18)32488-7 
 
Rezza G, Nicoletti L, Angelini R, Romi R, Finarelli AC, et al. (2007) Infection with chikungunya 
virus in Italy: an outbreak in a temperate region. Lancet 370:1840-1846 
Schilte C, Staikowsky F, Couderc T, Madec Y, Carpentier F, Kassab S, Albert ML, Lecuit M, 
Michault A. Chikungunya virus-associated long-term arthralgia: a 36-month prospective 
longitudinal study. PLoS Negl Trop Dis. 2013;7(3):e2137. 
Simon F, Parola P, Grandadam M, Fourcade S, Oliver M, Brouqui P, Hance P, Kraemer P, Ali 
Mohamed A, de Lamballerie X, Charrel R, Tolou H. Chikungunya infection: an emerging 
rheumatism among travelers returned from Indian Ocean islands. Report of 47 cases. 
Medicine (Baltimore). 2007 May;86(3):123-37. 
Soares de Araújo J, Regis CT, Silva Gomes RG, et al. Microcephaly in northeast Brazil: a 
review of 16 208 births between 2012 and 2015. http://who.int/bulletin/online_first/16-
170639.pdf?ua=1 (accessed Feb 8, 2016).  
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 81/82 
Staples JE, Breiman RF, Powers AM. Chikungunya fever: an epidemiological review of a re-
emerging infectious disease. Clin Infect Dis. 2009 Sep 15;49(6):942-8. 
Stebbings R, Février M, Li B, Lorin C, Koutsoukos M, Mee E, et al. Immunogenicity of a 
recombinant measles-HIV-1 clade B candidate vaccine. PLoS One. 2012;7:e50397.  
Tsetsarkin KA, Vanlandingham DL, McGee CE, Higgs S (2007) A single mutation in 
chikungunya virus affects vector specifity and epidemic potential. PLoS Pathog 3: e201. 
doi:10.1371/journal.ppat.0030201 
U.S. Department of Health and Human Services, Food and Drug Administration, Center for 
Biologics Evaluation and Research, September 2007: Guidance for Industry: Toxicity Grading 
Scale for Healthy Adult and Adolescent Volunteers Enrolled in Preventive Vaccine Clinical 
Trials; 
see 
http://www.fda.gov/BiologicsBloodVaccines/GuidanceComplianceRegulatoryInformation/Guid
ances/Vaccines/ucm074775.htm 
Vazeille M, Moutailler S, Coudrier D, Rousseaux C, Khun H, et al. (2007) Two Chikungunya 
isolates from the outbreak of La Reunion (Indian Ocean) exhibit different patterns of infection 
in the mosquito, Aedes albopictus. PLoS ONE 2: e1168. doi:10.1371/journal.pone.0001168 
Wang E, Volkova E, Adams AP, Forrester N, Xiao SY, et al. (2008) Chimeric alphavirus 
vaccine candidates for chikungunya. Vaccine 26: 5030-5039 
WMA Declaration of Helsinki - Ethical Principles for Medical Research Involving Human 
Participants; JAMA. 2013;310(20):2191-2194.  
WHO. Guillain-Barré syndrome – El Salvador. Jan 21, 2016. http://www.who.int/csr/don/21-
january-2016-gbs-el-salvador/en/ (accessed Feb 5, 2016).  
WHO, Media centre, 2016: See: http://www.who.int/mediacentre/news/notes/2016/research-
development-zika/en/ 
Zanluca C et al, . Mem Inst Oswaldo Cruz 2015; 110: 569–72.  
 
06CCVP
06G6F2

MV-ZIKA-RSP-101 
    
 
EudraCT 2019-000840-93                       Protocol 
Version 2.1, 28 September 2020 
 
 
 
 
 
Page 82/82 
22. APPENDICES 
 
Appendix 1: Template distributed to participants to assess local reaction 
 
 
 
06CCVP
06G6F2

&&93
PPD
PPD
*)

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 1 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
 
 
Statistical Analysis Plan 
Preliminary and Final Analysis 
 
 Observer blinded, randomized trial to evaluate safety and 
immunogenicity of a novel vaccine formulation MV-ZIKA-RSP 
 
Protocol: MV-ZIKA-RSP-101 / EudraCT 2019-000840-93 
 
 
Confidential 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Sponsor: 
Themis Bioscience GmbH 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 2 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
Table of Contents 
 
1. 
OVERVIEW 
6 
 
Study Objectives 
6 
 
Primary Objective 
6 
 
Secondary Objectives 
6 
 
Study Design 
6 
 
Endpoints 
7 
 
Primary Endpoint 
7 
 
Secondary Endpoints 
7 
 
Sample Size Calculation 
7 
 
Study Flowchart and Study Procedures 
8 
2. 
GENERAL CONSIDERATIONS 
10 
 
Conduct of Analysis 
10 
 
Statistical Software and Quality Control 
10 
 
Applicable Standard Operation Procedures 
10 
 
Blinding and Randomization 
11 
 
Descriptive Analyses 
12 
 
Inferential Analyses 
12 
 
Center and Country Effect 
13 
 
Handling Missing Data 
13 
 
Protocol Deviations 
13 
 
Medical Coding 
14 
 
Analysis Populations 
14 
 
Safety Population 
14 
 
Modified Intent-to-Treat (mITT) Population 
14 
 
Per-Protocol (PP) Population 
15 
 
Subject Data Listings 
15 
 
Columns in Tables 
15 
 
Changes in the Conduct of the Study or Planned Analysis 
15 
3. 
OVERALL STUDY INFORMATION 
16 
 
Data points 
16 
 
Derivations and Definitions 
16 
4. 
BASELINE EVALUATION 
16 
 
Data points 
16 
 
Derivations and Definitions 
17 
5. 
IMMUNOGENICITY ANALYSIS 
17 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 3 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Data points 
17 
 
Derivations and Definitions 
17 
6. 
SAFETY ANALYSIS 
18 
 
Extent of Exposure 
18 
 
Data points 
18 
 
Derivations and Definitions 
19 
 
Adverse Events 
19 
 
Data points 
19 
 
Derivations and Definitions 
19 
 
Laboratory Parameters 
20 
 
Data points 
20 
 
Derivations and Definitions 
21 
 
Other Safety Parameters 
21 
 
Data points 
21 
7. 
LIST OF TABLES, DATA LISTINGS AND FIGURES 
23 
 
List of Tables 
23 
 
List of Data Listings 
29 
8. 
REFERENCES 
33 
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 4 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
Document 
Study 
MV-ZIKA-RSP-101 
Document 
Statistical Analysis Plan (SAP) 
Version 
Final 3.0 
Date 
24-Sep-2020 
Revision History 
Version 
Date 
Reason for Revision 
Final 1.0 
24-Mar-2020 
First version; 
Final 2.0 
10-Sep-2020 
Immunogenicity analysis performed at final analysis 
instead of preliminary analysis since laboratory results 
are not available for preliminary analysis; 
Final 3.0 
24-Sep-2020 
Replaced PRNT50 with VNT to be consistent with updated 
CSP (Version 2.1);  
Approval 
Name 
Role 
Date 
Signature 
Author 
Biostatistician 
 
 
Assign DMB 
Senior Biostatistician 
 
 
Themis Bioscience GmbH 
Clinical Project Manager 
 
 
 
 
 
 
 
 
 
 
 
Any photocopies taken of this document are not authorized or version controlled. 
*1
PPD
PPD
PPD
::.

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 5 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
List of Abbreviations 
 
AE 
Adverse Event 
ALT 
Alanine Aminotransferase  
ANOVA 
Analysis of Variance 
aPTT 
Activated Partial Thromboplastin Time 
AST 
Aspartate Aminotransferase  
ATC 
Anatomical Therapeutic Chemical 
BDRM 
Blind Data Review Meeting 
CI 
Confidence Interval 
CSP 
Clinical Study Protocol 
CSR 
Clinical Study Report 
DRM 
Data Review Meeting 
eCRF 
Electronic Case Report Form 
ELISA 
Enzyme Linked Immunosorbent Assay  
GMFI 
Geometric Mean Fold Increase 
GMT 
Geometric Mean Titer 
HIV 
Human Immunodeficiency Virus 
IMP 
Investigational Medicinal Product  
IMP 
Investigational Medical Product 
LLOQ 
Lower Limit of Quantification 
mITT 
Modified Intent-to-Treat 
PP 
Per-Protocol 
PRNT50 
Plaque Reduction Neutralization Test 50 % 
PT 
Prothrombin Time 
SOP 
Standard Operating Procedure 
SPS® 
Stabilizing and Protecting Solutions  
TLF 
Tables Listings and Figures 
ULOQ 
Upper Limit of Quantification 
VNT 
Virus Neutralization Test 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 6 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
1. 
OVERVIEW 
 
Study Objectives 
 
Primary Objective 
 
To investigate safety and tolerability of MV-ZIKA-RSP, a novel liquid vaccine formulation, up to day 56 after 
first immunization consisting in one or two vaccinations 
Safety and tolerability will be assessed by the rate adverse events (AEs) after the first vaccination up to day 56. 
 
Secondary Objectives 
 
To investigate long-term safety, up to study day 182, after first immunization 
Long term safety and tolerability will be assessed by means of a phone call where participants will be asked about 
any adverse event (solicited, unsolicited or serious adverse event) experimented after the Visit 3 (day 56)  
 
 
To investigate immunogenicity up to day 56 after first immunization consisting in one or two vaccinations 
with different doses 
To cover this objective the presence of functional anti-zika antibodies will be determined by means VNT and ELISA 
at the following time points: Visit 1 (day 0), Visit 2 (day 28) and Visit 3 (day 56) after first vaccination 
 
To get information about the optimal dose of MV-ZIKA-RSP vaccine regarding safety, tolerability and 
immunogenicity 
An evaluation of the optimal dose will be determined after analysis of the safety, tolerability and immunogenicity 
(VNT) 
 
To investigate specific cell mediated immunity induced by all different treatment groups 
Cell mediated immunity up to day 56 will be assessed by T cell analysis of PBMC isolated on study days 0, 28 and 
56 
 
Study Design 
This is an observer-blinded, block-randomised, dose finding, phase I trial, comparing different dose levels of MV-
ZIKA-RSP to evaluate safety, tolerability and immunogenicity, of this novel ZIKA-RSP vaccine. Placebo (physiological 
saline solution) will be applied to blind the different treatment schedules. 
The study will be conducted at a single study site, Medical University Vienna. 
After the screening procedures, 48 healthy male and female volunteers aged 18-55 years will be randomly assigned 
to one of four treatment groups (A, B, C or D). Participants will be assessed for immunogenicity on days 0, 28 and 
56 (treatment period), as confirmed by the presence of functional anti-zika-rsp antibodies determined by (VNT) and 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 7 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
by ELISA, at the same time safety will be also assessed. After the treatment period, participants will be call by phone 
(day 182) for evaluation of safety follow-up.  
Study duration per subject will be 182 days (6 months), consisting of 4 weeks treatment period (study day 0 till 
study day 28) and 22 weeks follow up period (study day 2 till study day 182) 
 
Endpoints 
 
Primary Endpoint 
 
Rate of adverse events (AEs) up to study day 56 
 
Secondary Endpoints 
 
Rate of solicited and unsolicited AEs, as well as, reported serious adverse events (SAEs) up to study day 
182 (long-term safety) compared between the 4 different treatment groups.  
 
Immunogenicity on study days up to study day 56 as confirmed by the presence of functional anti-zika 
antibodies as determined by VNT and by ELISA 
 
Cell-mediated immunity specific for ZIKA-RSP, up to study day 56, confirmed by the presence of specific 
functional CD4+ and CD8+ T-cells 
 
Safety laboratory parameters (haematology, serum chemistry, urinalysis) 
 
Sample Size Calculation 
A formal sample size calculation was not conducted. The sample size of 48 participants has been determined based 
on prior experience in evaluating the safety and immunogenicity of vaccines and is typical for early phase clinical 
studies. Sample size for this study was determined on grounds of feasibility and common practice in similar trials. 
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 8 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Study Flowchart and Study Procedures 
Study flowchart: 
 
Study procedures:  
 
 
Screenin
g 
 
Treatment 
 
Post Treatment 
Safety Follow 
Up 
ET 
Visit 
VS 
V1 
Post 
dose 
safety FU 
V2 
V3 
 Primary 
Endpoint 
V4 
(1) 
Study Day 
-14 to -1 
0 
 
28±
3 
56 ±3 
182 ±14 
 
Informed Consent 
x 
 
 
 
 
 
 
Inclusion/Exclusion Criteria 
x 
x 
 
 
 
 
 
Medical History 
x 
x 
 
 
 
 
 
Vaccination History  
x 
x 
 
 
 
 
 
Vital Signs (2) 
x 
x 
 
x 
x 
 
x 
Symptom-directed Physical Examination 
 
x 
 
x 
x 
 
x 
System-based Physical Examination (6) 
x 
x  
 
x 
x  
 
x  
Adverse Events (3) (7) 
 
x  
x 
x 
x 
x 
x 
Prior/Concomitant Medication (4) 
x 
x 
x 
x 
x 
x 
x 
Clinical Laboratory (5) 
x 
 
 
x 
x 
 
x 
Randomization 
 
x 
 
 
 
 
 
Urine Pregnancy Test (12) 
x 
x 
 
x 
x 
 
x 
Study Treatment  
 
x 
 
x 
 
 
 
Immunogenicity (VNT) (12) 
 
x 
 
x 
x 
 
x 
T-cell immunity (PBMCs) (12) 
 
x 
 
x 
x 
 
x (11) 
Zika antibody (ELISA) (12) 
 
x 
 
x 
x 
 
x 
Measles antibody (ELISA) (12) 
 
x 
 
x 
x 
 
x 
Local Tolerability (7) 
 
x 
 
x 
x 
 
x 
Dispense subject diary 
 
x 
 
x 
 
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 9 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
Collect and review subject diary 
 
 
 
x 
x 
 
x 
Safety phone call (9) 
 
 
x (9) 
 
 
x (10) 
 
 
(1) 
Early termination visit should – if possible - be conducted 28±3 days after last IMP administration. 
(2) 
Systolic/diastolic blood pressure, pulse rate, body temperature 
(3) 
Conditions/symptoms noted prior to randomization should not be reported as AE, but should be reported as medical history 
(4) 
Medications administered up to 30 days prior to screening until end of study participation should be documented 
(5)  
The clinical laboratory comprises: Hematology (hemoglobin, hematocrit, erythrocyte count, differential white blood count, 
platelets), Chemistry (creatinine, potassium, sodium, calcium, AST, ALT, alkaline phosphatase, bilirubin), Coagulation 
(prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen), Urinalysis (standard urine test stick for 
determining pH-value, glucose, protein, bilirubin, urobilinogen, red blood cells, white blood cells, nitrite, ketone and specific 
gravity), Virology (only at screening; HBs-Ag, anti-HBc-Ab, anti-HCV-Ab, HIV 1/2 Ab/HIV-1 Antigens if not done within 30 days 
before screening) 
(6) 
To be performed if necessary, according to findings in symptom-directed physical examination 
(7) 
To be performed 1 hour after each IMP administration and additionally at the subsequent visit (prior to IMP administration, if 
applicable) 
(8) 
Participants should be interviewed regarding potential AEs since the last study visit. If indicated, participants should be called in 
for an unscheduled visit. 
(9) 
The post dose safety call follows up should be conducted 7±1 days (one week) after the first vaccination. 
(10) 
The post treatment safety call follows up should be conducted 182 days (6 months) after the first study treatment also in case of 
an early termination. 
(11) 
The amount of blood sample taken during the early termination (ET) visit depends on the time point of the visit. If the ET-visit 
replaces the primary endpoint visit on day 56±3, 48 to 60 mL blood should be taken; otherwise 24 to 48 mL are enough. 
(12) 
Urine for pregnancy test should be taken before vaccination 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 10 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
2. 
GENERAL CONSIDERATIONS 
 
Conduct of Analysis 
A preliminary analysis including safety and immunogenicity data will be performed after all participants completed 
Visit 3 (day 56) and therefore terminated the treatment period of the study. This analysis will cover the primary 
safety endpoint (adverse events up to day 56) and some immunogenicity endpoints. The purpose of this analysis is 
to provide highly valuable data for further zika vaccine development.  
Results from this preliminary analysis will not have any impact on the study design or the participants during the 
follow up period of the study. 
 
The final analysis will be conducted after the last subject completed the Post Treatment Safety Follow-Up and the 
database was closed. All data of the study will already be available at the preliminary analysis except the Adverse 
Events and Prior/Concomitant Medication data from Visit 4 (Day 182). Thus, only Adverse Events and 
Prior/Concomitant Medication data will be analyzed at the final analysis. If unexpected changes occur e.g. in the 
immunogenicity data, analyses may also be repeated at the final analysis. 
 
SAP update (Final 2.0): Due to the covid pandemic, at the time of analysis of the samples, laboratory capacities 
were limited, and no results were available. For this reason, immunogenicity analysis was not conducted for the 
preliminary analysis. Instead, the immunogenicity analysis will be done as described in Section 5 for the final 
analysis only. 
 
Statistical Software and Quality Control 
All statistical analyses will be performed using SAS® version 9.3 or higher. Tables, figures and data listings will be 
generated in Microsoft® Word® as well as PDF® format. 
Quality control of SAS® programs will include a review of the whole process of result generation: 
 
Review of all analysis SAS® programs 
 
Review of SAS® log for errors, warnings and other notes that could indicate mistakes in the programs 
 
Review of all tables, listings and figures for completeness and correctness 
 
Applicable Standard Operation Procedures 
The applicable Standard Operating Procedures (SOPs) of Assign DMB for this study are: 
STAT01 
Statistical Analysis File 
STAT03 
Statistical Analysis Plan 
STAT04 
Interim Analysis 
STAT05 
Randomization and Unblinding 
STAT06 
Data Review Meeting 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 11 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
STAT07 
Report Writing 
SAS01 
SAS General Principles 
SAS04 
Handling of Statistical Analyses 
 
Blinding and Randomization 
Blinding: 
As this study will be conducted in an observer-blinded manner, unblinded study team members responsible for 
randomisation, monitoring (unblinded monitor) and preparation of IMP, will otherwise not be involved in the conduct 
of the study. 
All participants, the investigator and site personnel performing study related assessments, as well as the sponsor’s 
representatives involved in the monitoring (blinded monitor) and conduct of the study, will be blinded to treatment 
assignment. 
Besides one set of randomisation envelopes, the site will receive an additional set of emergency envelopes stored 
in the ISF for unblinding purposes. As the knowledge of the subject’s treatment might become mandatory (e.g. in 
case of emergency) the investigator has the possibility to open the emergency envelope, identified with the subject’s 
randomisation number, and unblind the subject.  
The investigator and site personnel assessing AEs, all participants, as well as one of the sponsor’s representatives 
involved in the monitoring and conduct of the study will be blinded to which vaccine was administered. Only the 
unblinded monitor, site personnel performing randomisation and preparation of IMP will be unblinded. 
The primary endpoint as well as most of the secondary endpoints will be covered in the preliminary analysis after 
Day 56 and only a safety follow-up phone call will be performed after Day 56. The sponsor personnel and ADMB 
team, involved in generation and review of the preliminary analysis and underlying data management activities, will 
be unblinded after the Blind Data Review Meeting (Section 2.9) and database snapshot for preliminary analysis. 
 
Randomization:  
At Visit 1 (day 0) 48 eligible participants will be randomly assigned to one of four treatment groups (A, B, C or D) by 
means of randomisation envelopes provided by data management.  
Each subject will be assigned a unique three-digit randomisation number in ascending order, by opening the 
randomisation envelope with the lowest free (unassigned) randomisation number available. Randomisation should 
be performed as late as possible, i.e. when knowledge of the assigned treatment group becomes necessary. Only 
trained members of the unblinded study team will be authorised to open one envelope per subject, containing the 
information about the allocated treatment group. 
 
The 48 participants will be distributed between the 4 groups using a randomisation ratio of 14 : 14 : 12 : 8 = A : B : 
C : D, with an adequate representation of high dose, low dose and placebo subjects (2 : 2 : 0 : 2 = A : B : C : D) within 
the first 6 subjects to allow for the safety evaluation after the first vaccination. 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 12 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
A subject will be considered enrolled once the first IMP administration has been performed according to 
randomisation. 
 
Descriptive Analyses 
Descriptive analyses of continuous variables (summary statistics) will be described with the number of non-missing 
observations, arithmetic mean, standard deviation (±SD), median, quartiles (Q1 and Q3) and range (minimum and 
maximum). 
Descriptive analysis of continuous immunogenicity variables (i.e. tables for the GMT and GMFI) will be described 
with the number of non-missing observations, geometric mean, confidence intervals for the geometric mean, 
standard deviation of logarithmic values, median, quartiles (Q1 and Q3) and range (minimum and maximum). 
Categorical variables (frequency statistics) will be described with the number of non-missing observations and 
percentages (%). Percentages will be calculated within each stratum on the total number of non-missing 
observations, if not stated otherwise. 
 
Inferential Analyses 
The analysis for primary endpoint will compare adverse events rates up to day 56 between treatment groups using 
Fisher-Freeman-Halton test.  The same overall test between treatment groups will be performed for all adverse event 
tables in the Day 56 analysis as well as the final analysis. 
 
The immunogenicity analysis will compare the anti-zika-rsp VNT antibody geometric mean titer (GMT) in the Per 
Protocol (PP) analysis population between the treatment groups. GMTs and GMT ratios will be estimated by applying 
an analysis of variance including the factor treatment group. This will be done using log10 transformed data and 
taking the anti-log of the resulting point estimates for the least squares means, least squares means differences 
and the corresponding 2-sided 95% CIs.  
 
P-values will also be provided to compare GMTs between the treatment groups adjusted for multiple comparisons 
according to Tukey-Kramer.  
 
For the primary immunogenicity analysis, values below the lower limit of detection are replaced by half of the limit 
and values above the upper limit are replaced by the upper limit. 
As sensitivity analysis, an analysis of immunogenicity will be applied, that does not use such imputation. Therefore, 
observations will be interval-censored and GMTs will be estimated by assuming normal distribution for the log-
transformed values (intervals). Values below the lower limit of detection will be left-censored at the lower limit and 
values above the upper limit (if applicable) will be right-censored at the upper limit.  
 
Seroconversion rates and positivity rates will be compared between groups using Fisher-Freeman-Halton test. 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 13 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Center and Country Effect 
Not applicable since this is a single-center study. 
 
Handling Missing Data 
Generally, missing values will not be imputed, and the analysis will be limited to observed values. But for missing 
data in the AE (e.g. missing information about seriousness, severity or causality) a worst case approach will be 
applied. For example, adverse events with missing severity assessments will be considered as “Potentially life 
threatening” and events with missing relationship to study treatment are counted as “Definitely” related to study 
treatment. In case of missing assignment to solicited or unsolicited, this AE will neither be counted in tables for 
solicited AEs nor in tables for unsolicited AEs but in tables for all AEs. 
 
 Protocol Deviations 
Protocol deviations will be collected only for the preliminary analysis. A listing of the potential protocol deviations 
will be prepared by Assign Data Management and Biostatistics GmbH using information from the eCRF 
documentation and monitoring reports. These protocol deviations will be classified into major or minor protocol 
deviations based on their possible impact on the study results in a Blind Data Review Meeting (BDRM) prior to the 
preliminary analysis attended by Assign Data Management GmbH and Themis Bioscience GmbH (sponsor). 
Major protocol deviations will include but are not limited to the following: 
• 
Subjects with any confirmed immunosuppressive or immunodeficient condition, including human 
immunodeficiency virus (HIV), hepatitis A, B or C infection or a family history of congenital or 
hereditary immunodeficiency 
• 
Subjects who received the wrong or no IMP  
• 
Subjects who received less than the protocol-defined number of vaccinations 
• 
Subjects who received a wrong dose other than they were randomized to 
• 
Subjects who entered the study even though they did not satisfy the entry criteria 
• 
Subjects with substantial time window violations on vaccination visits 
• 
Subjects who received a forbidden prior or concomitant medication 
o 
Immunosuppressive drugs: Use of corticosteroids (excluding topical preparations) or 
immunosuppressive drugs within 30 days prior to first IMP administration, or anticipated use 
during the trial 
o 
Treatments with non-study licensed vaccines within 4 weeks prior to first study vaccination until 
the end of the treatment period (day 56) 
o 
Receipt of blood products or immunoglobulins within 120 days prior to the Screening Visit or 
anticipated receipt of any blood product or immunoglobulin before completion of the treatment 
period (day 56) 
• 
Further protocol deviations that could impact immune response that have not been described above  
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 14 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
These criteria for potential protocol violation are identified at the time of planning the study. However, during the 
course of the trial unforeseen events may occur or new scientific knowledge may become available, therefore final 
decisions on whether any protocol violation could impact immune response and thus lead to exclusion from the per-
protocol (PP) population will be made by the sponsor on a case by case basis in a blinded manner (and prior to study 
unblinding). 
All protocol deviations will be listed in the analysis and described in the Clinical Study Report (CSR). 
In case of unexpected changes or further protocol deviations documented after the BDRM, it may be discussed in 
a second DRM prior to database closure which analyses may be repeated for the final analysis.  
 Medical Coding 
Adverse events, medical history and concomitant procedures will be coded using MedDRA in the most current 
version. Concomitant medications and vaccination history will be coded using WHO Drug Reference List and 
Anatomical Therapeutic Chemical (ATC) Classification System in the most current version. The used dictionary 
versions will be indicated in the respective tables and listings. 
 Analysis Populations 
 Safety Population 
All safety analyses will be based on the safety population, defined as participants who entered into the study and 
received at least one IMP administration. All analysis based on the Safety Population will be carried out using the 
actual treatment received. 
Possible issues with wrong treatment and assignment of subjects to actual treatment group will be discussed in the 
DRM. In general, this assignment will be done in a conservative way to not bias the safety results towards MV-ZIKA-
RSP. If a subject in the high dose group receives a low dose, the subject will be analysed in the low dose group, for 
example.  
 Modified Intent-to-Treat (mITT) Population 
The secondary immunogenicity analyses will be based on the modified ITT population. The modified intent-to-treat 
(mITT) analysis population is defined to include all participants randomised who receive at least one IMP 
administration. Participants will be analysed according to the treatment group they had been allocated to, rather 
than by the actual treatment they received. 
When no subjects received wrong treatment, mITT and Safety population do not differ and tabulations will only be 
done for safety population.  
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 15 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 Per-Protocol (PP) Population 
The immunogenicity analysis will be assessed primarily on the per-protocol (PP) population. The PP population 
includes participants without protocol deviations that could impact immune response. Examples that may lead to 
exclusion from the PP population are provided here: 
- 
Immunosuppressive drugs: Use of corticosteroids (excluding topical preparations) or immunosuppressive 
drugs within 30 days prior to first IMP administration, or anticipated use during the trial. 
- 
Participants with any confirmed immunosuppressive or immunodeficient condition, including human 
immunodeficiency virus (HIV), hepatitis A, B or C infection or a family history of congenital or hereditary 
immunodeficiency 
- 
Participants who received the wrong or no IMP 
These criteria for potential protocol violation are identified at the time of planning the study. However, during the 
course of the trial unforeseen events may occur or new scientific knowledge may become available, therefore final 
decisions on whether any protocol violation could impact immune response and thus lead to exclusion from the PP 
population will be made by the sponsor on a case by case basis in a blinded manner (and prior to study unblinding 
at the preliminary analysis). Sample testing issues may also lead to exclusion from the PP population for particular 
time points.  
 Subject Data Listings 
All treated subjects will be included in listings if not stated otherwise (e.g. subject overview listings, screening failure 
listing). Data listings will include the subject number as identifier (and parameter and/or visit if available) and will 
be sorted by treatment group, subject number and parameter and/or visit if available (in this order). Columns that 
indicate the treatment group will be shown in all listings. 
 Columns in Tables 
Every treatment group has its own column in the analysis and an additional total column will be added. For OV 
tables, where all subjects are shown, there is an additional column with subject not randomized.  
 Changes in the Conduct of the Study or Planned Analysis 
No changes to the statistical analysis as compared to CSP are planned. 
 
 
 
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 16 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
3. 
OVERALL STUDY INFORMATION 
For visit attendance and study status the safety population is used. For the other OV tables all subjects are used. 
 
Data points 
The following information will be analyzed descriptively and corresponding details on the subject level will be 
provided in data listings: 
 
Subject overview (screened, randomized, 1st/2nd IMP administered, study populations…) 
 
Visit attendance 
 
Protocol deviations 
 
Study status (incl. drop-outs and reason) 
 
Screening failures 
For details on table and listing content, see Section 7. 
 
Derivations and Definitions 
Details about study population can be found in Section 2.11. 
4. 
BASELINE EVALUATION 
Baseline analysis will be presented for the Safety, mITT population and for the PP population. In case populations 
do not differ, tabulations will not be repeated. 
 
Data points 
The following information will be analyzed descriptively and corresponding details on the subject level will be 
provided in data listings: 
 
Demographic information (age, sex, race) 
 
System-based Physical examination (will be listed only) 
 
Vaccination history (by ATC level 3) 
 
Virology (will be listed only) 
 
Medical history 
 
Prior and concomitant medications 
 
Pregnancy Test (will only be listed) 
 
Concomitant Procedure (will only be listed) 
 
Vital Signs at screening 
For details on table and listing content, see Section 7. 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 17 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Derivations and Definitions 
Medications stopped clearly prior (<) to Day 0 (Visit 1) will be considered prior medications, all other medications 
are considered to be concomitant. Medications with a missing or incomplete stop date where it cannot clearly be 
decided if the stop date was before or after Day 0 will be considered concomitant. 
5. 
IMMUNOGENICITY ANALYSIS 
The immunogenicity analysis will be performed for the mITT and PP population. All tables will be presented by 
treatment group and overall. Data from the early termination visit or unscheduled visits will be listed only and will 
not be shown in summary tables by time point. 
 
Data points 
The following information will be analyzed by visit descriptively in tables and corresponding details on the subject 
level will be provided in data listings:  
 
GMT of Zika-RSP VNT antibody (incl. ANOVA with fixed treatment group as described in Section 2.6) 
 
Seroconversion rate for Zika-RSP VNT antibody (incl. test as described in Section 2.6) 
 
Positive/Negative results for NT 
 
GMT of Zika-RSP-ELISA antibodies (incl. ANOVA with fixed treatment group) 
 
Seroconversion rate for Zika-RSP-ELISA antibodies  
 
Positive/Negative results for ZIKA ELISA 
 
GMT of Measles-ELISA antibodies  (incl. ANOVA with fixed treatment group) 
 
Seroconversion rate for Measles-ELISA antibodies 
 
Positive/Negative results for MV ELISA 
 
specific functional CD4+ T-cells by visit (without geometric mean): 
o 
E+prM positive/negative 
o 
E values 
o 
prM values 
Sampling details for VNT, ELISA Zika-RSP, ELISA measles and T-cells immunity (will be listed only) 
Specifications on TLFs are provided in Section 7. 
 
Derivations and Definitions 
 
For final analysis all time points, including Visit 1 (Day 0), Visit 2 (Day 28) and Visit 3 (Day 56) will be 
analyzed. 
 
Seroconversion rate for anti-zika-rsp VNT titers is defined as proportion of subjects achieving anti-zika-rsp 
VNT titers ≥ 20 at a certain time point. 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020)
Adapted from:
Page 18 of 33
STAT03_A Statistical Analysis Plan
Version 6.0, Effective Date 16-Feb-2018
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB.
x
Seroconversion rate for anti-zika-rsp ELISA titers is defined as 4-fold increase over baseline (Visit 1 –Day 
0). If no Visit 1 (Day 0) sample is available, no seroconversion will be determined. 
x
For MV ELISA, Zika ELISA, NT and T-Cell an analysis of positive versus negative values is planned. Since 
such a definition could not be discussed and communicated with the central laboratory at the time of 
generation of this SAP, details about used limits will be shown as footnotes in the corresponding tables and 
explained in the Clinical Study Report.
x
For the different immunogenicity parameter there are different lower limit of quantification (LLOQ).
o
In data listings, the original reported value “<LLOQ” will be displayed.
o
For calculations (GMT, median…) the half of the true LLOQ value will be used for such values. 
o
If another unexpected value is reported as LLOQ in the data than mentioned here, the half value 
of the reported LLOQ will be used for the calculations. 
x
For the different immunogenicity parameters there are different upper limits of quantification (ULOQ)
o
In data listings, the original reported value “>ULOQ” will be displayed.
o
For calculations (GMT, median…) the ULOQ value will be used for such values.
o
If another unexpected value is reported as ULOQ in the data than mentioned here, this ULOQ value
will be used for calculations. 
x
For analysis using interval-censored data an interval will be defined for every observation. Observations 
between LLOQ and ULOQ will be handled as interval with left limit and right limit being the observed 
value. For values <LLOQ the interval will be left-censored with the LLOQ as right limit.  For values >ULOQ 
the interval will be right-censored with the ULOQ as left limit.  
6.
SAFETY ANALYSIS 
All safety analyses will be performed for the safety population. 
Extent of Exposure
Data points
The following information will be analyzed descriptively and corresponding details on the subject level will be 
provided in data listings:
x
Treatment duration
x
Study duration
x
Study vaccine administration (will be listed only)
For details on table and listing content, see Section 7.
*1
CCI
CCI
CCI
CCI
::.

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 19 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Derivations and Definitions 
 
Treatment duration [days] = date of last administered vaccine – date of first vaccination +1 (only if more 
than one vaccine administered) 
 
Study duration [days] = date of last attended visit (or date of study discontinuation, if applicable) – date 
of signed informed consent + 1 
 
Adverse Events 
 
Data points 
The following analyses will be provided for all Adverse Events: 
 
Adverse event overview for solicited and unsolicited events (e.g. any AE, any SAE, any related AE).  
 
Solicited Adverse Events by symptom 
 
Solicited Adverse Events by symptom and severity 
 
Unsolicited Adverse Events by SOC and PT, overall and for specific types of Adverse Events (e.g. SAEs, 
related AEs) 
 
Unsolicited Adverse Events by severity and for specific types of Adverse Events 
 
Unsolicited Adverse Events by causality and for specific types of Adverse Events 
For details on table and listing content, see Section 7. 
 
Derivations and Definitions 
 
All solicited and unsolicited Adverse Events up to Day 56 (Visit 3) will be analyzed for the preliminary 
analysis and all AE in the database for the Final Analysis. The following rules will be applied to identify AEs 
up to Day 56 for preliminary analysis: In general, all AE with start date prior to or at Visit 3 are considered. 
If a subject has no Day 56 visit (Visit 3) then all AEs within 56 days from Visit 1 (Day 0) (Visit 1 + 56 days) 
are included. If start date of AE is incomplete (e.g. missing start day) and the AE could theoretically by within 
Visit 3, or if Visit 3 is not available, within 56 days from Visit 1 (Visit 1 + 56 days), it will be handled as AE 
up to Day 56 (Visit 3) and thus be included in the preliminary analysis. If start date of AE is unknown it will 
be handled as AE up to Day 56 visit (Visit 3) and thus be included in the preliminary analysis.  
 
Percentages for AE rates are based on N (treatment group totals). 
 
An Adverse Events is considered to be related to study treatment if “Relationship to Study Treatment” is 
answered with “definitely”, “probably” or “possibly” or the causality is missing. 
 
Tables showing “severe” events will include events with grade 3 or 4 or missing grade. 
 
In tables showing subjects with AEs by maximum severity, subjects will be counted only in the highest 
grading category (=”potentially life threatening”), but events will be counted in each reported grading 
category. 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 20 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
In tables showing AEs by causality, subjects will be counted only in the strongest relationship category 
(=”definitely”), but events will be counted in each category. 
 
Laboratory Parameters 
Data from the early termination visit or unscheduled visits will be listed only and will not be shown in summary tables 
by time point. Laboratory data from scheduled visits (VS, Visit 2 and Visit 3) will be tabulated by time point. 
Specifications on TLFs are provided in Section 7: 
For details on table and listing content, see Section 7. 
 
Data points 
 
Hematology 
o 
Hemoglobin 
o 
Hematocrit 
o 
Erythrocytes 
o 
Leukocytes 
o 
Neutrophils 
o 
Lymphocytes 
o 
Monocytes 
o 
Eosinophils 
o 
Basophils 
o 
Platelets 
 
Clinical Chemistry 
o 
Creatinine 
o 
Potassium 
o 
Sodium 
o 
Calcium 
o 
Aspartate Aminotransferase 
o 
Alanine Aminotransferase 
o 
Alkaline Phosphatase 
o 
Bilirubin 
 
Coagulation 
o 
Prothrombin Time 
o 
Activated Partial Thromboplastin Time 
o 
Fibrinogen 
 
Urinalysis 
o 
Specific gravity 
o 
pH 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 21 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
o 
Leukocytes 
o 
Nitrite 
o 
Protein 
o 
Glucose 
o 
Ketone 
o 
Urobilinogen 
o 
Bilirubin 
o 
Erythrocytes 
The following parameters will be analyzed descriptively by time point: 
 
Absolute values (summary statistics for hematology, clinical chemistry and coagulation) 
 
Absolute change from baseline (Screening) (summary statistics for hematology, clinical chemistry and 
coagulation) 
 
Number of subjects with values above/below normal range (for hematology, clinical chemistry and 
coagulation) 
 
Urinalysis results (summary statistics for quantitative parameters, frequency statistics for qualitative 
parameters) 
 
Derivations and Definitions 
 
No conversion of laboratory parameters will be performed. The parameters listed above will be tabulated 
and listed in the units reported by the study site.  
 
Absolute change from baseline will be calculated as difference from value of the current visit and 
baseline value (screening). 
 
Other Safety Parameters 
 
Data points 
Data from the early termination visit or unscheduled visits will be listed only and will not be shown in summary tables 
by time point. The following vital sign parameters will be summarized descriptively:  
 
Diastolic blood pressure 
 
Systolic blood pressure 
 
Pulse rate 
 
Temperature 
The following data will be listed only:  
 
Pregnancy test 
 
Local tolerability 
 
Local tolerability – post vaccination 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 22 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Symptom-directed physical examination 
 
System-based physical examination 
For details on table and listing content, see Section 7. 
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 23 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
7. 
LIST OF TABLES, DATA LISTINGS AND FIGURES 
 
List of Tables 
No. for prel. 
Analysis 
No. for FA 
Legend 
Content 
Overall Subject Information 
1.1.1.1 
2.1.1.1 
Subject Overview (All subjects) 
 
Randomized 
 
Safety Population 
 
mITT Population 
 
PP Population 
 
Number of subjects that received first 
vaccination 
 
Number of subjects that received 
second vaccination 
 
Number of subjects that missed any 
vaccination 
1.1.1.2 
2.1.1.2 
Protocol Deviations by Deviation 
Type (All subjects) 
 
Any major PD 
 
Any major PD by category 
 
Any minor PD 
Any minor PD by category 
1.1.1.3 
2.1.1.3 
Number of Screening Failures 
and Reasons 
 
Number of screening failures 
 
Reason for screening failure 
1.1.1.4 
2.1.1.4 
Subjects by Visit (Safety 
Population) 
 
1.1.1.5 
2.1.1.5 
Study Status (Safety Population) 
 
Visit attendance status 
 
Primary reason for early termination 
1.1.2.4-
1.1.2.5 
2.1.2.4-
2.1.2.5 
Repeat tables x.1.1.4-x.1.1.5 for 
mITT population (if not the same 
as Safety Population) 
 
1.1.3.4- 
1.1.3.5 
2.1.3.4- 
2.1.3.5 
Repeat tables x.1.1.4-x.1.1.5 for 
PP population 
 
NOTE: Overall Subject Information tables will also be provided for final analysis including all information reported 
during the study 
Baseline Evaluation 
1.2.1.1 
n/a 
Summary Table of Demographic 
Information (Safety Population) 
 
Age 
 
Gender 
 
Race 
 
Height [cm] 
 
Weight [kg] 
 
 
1.2.1.2 
n/a 
Vaccination History by and ATC 
Level 3 (Safety Population) 
 
1.2.1.3 
n/a 
Medical History Entry by SOC and 
PT (Safety Population) 
 
1.2.1.4 
2.2.1.4 
Prior Medications by ATC Level 2 
and 3 (Safety Population) 
 
1.2.1.5 
2.2.1.5 
Concomitant Medications by ATC 
Level 2 and 3 (Safety Population) 
 
1.2.1.6 
n/a 
Vital Signs at Baseline (Safety 
Population) 
 
Systolic blood pressure [mmHg] 
 
Diastolic blood pressure [mmHg] 
 
Pulse rate [beats/min] 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 24 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Temperature [C] 
1.2.2.1-
1.2.2.6 
2.2.2.4-
2.2.2.5 
For preliminary analysis repeat 
tables 1.2.1.1-1.2.1.6 and for 
final analysis tables 2.2.1.4-
2.2.1.5 for mITT population (if 
not the same as Safety 
Population). 
 
NOTE: Prior/Concomitant Medication tables will also be provided for final analysis but including all 
Prior/Concomitant Medications reported during the study 
Immunogenicity Analysis 
Zika-RSP VNT Antibody 
n/a 
2.3.1.1 
GMT of Zika-RSP VNT Antibody by 
Visit (mITT Population) 
 
n/a 
2.3.1.2 
GMT of Zika-RSP VNT Antibody 
based on interval-censored data 
by Visit (mITT Population) 
 
n/a 
2.3.1.3 
ANOVA for GMT of Zika-RSP VNT 
Antibody between treatment 
groups by Visit (mITT Population) 
 
n/a 
2.3.1.4 
Seroconversion rate for of Zika-
RSP VNT Antibody by Visit (mITT 
Population) 
 
n/a 
2.3.1.5 
Positivity of Zika-RSP VNT 
Antibody by time point 
 
Zika-RSP ELISA Antibody 
n/a 
2.3.1.6 
GMT of Zika-RSP-ELISA 
Antibodies by Visit (mITT 
Population) 
 
n/a 
2.3.1.7 
GMT of Zika-RSP-ELISA 
Antibodies based on interval-
censored data by Visit (mITT 
Population) 
 
n/a 
2.3.1.8 
ANOVA for GMT of Zika-RSP-
ELISA Antibodies between 
treatment groups by Visit (mITT 
Population) 
 
n/a 
2.3.1.9 
Seroconversion rate of Zika-RSP-
ELISA Antibodies by Visit (mITT 
Population) 
 
n/a 
2.3.1.10 
Positivity of Zika-RSP-ELISA 
Antibodies by time point 
 
Measles-ELISA antibodies 
n/a 
2.3.1.11 
GMT of Measles-ELISA Antibodies 
by Visit (mITT Population) 
 
n/a 
2.3.1.12 
GMT of Measles-ELISA Antibodies 
based on interval-censored data 
by Visit (mITT Population) 
 
n/a 
2.3.1.13 
ANOVA for GMT Measles-ELISA 
Antibodies between treatment 
groups by Visit (mITT Population) 
 
n/a 
2.3.1.14 
Seroconversion rate of Measles-
ELISA Antibodies by Visit (mITT 
Population) 
 
n/a 
2.3.1.15 
Positivity of Measles-ELISA 
Antibodies by time point 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 25 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
T cell immunity  
n/a 
2.3.1.16 
Median of SFC of T cell immunity-
PBMCs by Visit (mITT Population) 
 
 
n/a 
2.3.1.17 
Positivity of SFC of T cell 
immunity-PBMCs by time point 
 
n/a 
2.3.2.1-
2.3.2.17 
Repeat Immunogenicity tables 
2.3.1.1-2.3.1.17 for PP 
Population. 
 
Safety Results 
Extent of Exposure 
1.4.1.1 
2.4.1.1 
Extent of Exposure (Safety 
Population) 
 
Study duration 
 
Treatment duration 
Adverse Events 
1.4.1.2 
2.4.1.2 
Summary Table of solicited and 
unsolicited Adverse Events 
(Safety Population) 
Number of subjects with: 
 
Any AEs 
 
Any related AEs 
 
Any severe AEs 
 
Any related severe AEs 
 
Any SAEs 
 
Any related SAEs 
 
Any medically attended AEs 
 
Any related medically attended AEs 
 
Any AEs leading to second vaccination 
not administered 
1.4.1.3 
2.4.1.3 
Summary Table of solicited 
Adverse Events (Safety 
Population) 
 
Any solicited AEs 
 
Any related solicited AEs 
 
Any severe solicited AEs 
 
Any related severe solicited AEs 
 
Any solicited SAEs 
 
Any related solicited SAEs 
 
Any medically attended solicited AEs 
 
Any related medically attended 
solicited AEs 
 
Any solicited AEs leading to second 
vaccination not administered 
1.4.1.4 
2.4.1.4 
Summary Table of solicited local 
Adverse Events (Safety 
Population) 
 
Any solicited local AEs 
 
Any related solicited local AEs 
 
Any severe solicited local AEs 
 
Any related severe solicited local AEs 
 
Any solicited local SAEs 
 
Any related solicited local SAEs 
 
Any medically attended solicited local 
AEs 
 
Any related medically attended 
solicited local AEs 
 
Any solicited local AEs leading to 
second vaccination not administered 
1.4.1.5 
2.4.1.5 
Summary Table of solicited 
systemic Adverse Events (Safety 
Population) 
 
Any solicited systemic AEs 
 
Any related solicited AEs 
 
Any severe solicited AEs 
 
Any related severe solicited systemic 
AEs 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 26 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
Any solicited systemic SAEs 
 
Any related solicited systemic SAEs 
 
Any medically attended solicited 
systemic AEs 
 
Any related medically attended 
solicited systemic AEs 
 
Any solicited systemic AEs leading to 
second vaccination not administered 
1.4.1.6 
2.4.1.6 
Summary Table of unsolicited 
Adverse Events  (Safety 
Population) 
 
Any unsolicited AEs 
 
Any related unsolicited AEs 
 
Any severe unsolicited AEs 
 
Any serious unsolicited AEs 
 
Any medically attended unsolicited 
AEs 
 
Any related severe unsolicited AEs 
 
Any related serious unsolicited AEs 
 
Any related medically attended 
unsolicited AEs 
 
Any unsolicited AEs leading to second 
vaccination not administered 
1.4.1.7 
2.4.1.7 
Subjects with at least one 
Adverse Event classified by 
Maximum Severity (Safety 
Population) 
 
Solicited Adverse Events 
1.4.1.8 
2.4.1.8 
Subjects with solicited local 
Adverse Events by Symptom 
(Safety Population) 
 
1.4.1.9 
2.4.1.9 
Subjects with solicited systemic 
Adverse Events by Symptom 
(Safety Population) 
 
1.4.1.10 
2.4.1.10 
Subjects with solicited Adverse 
Events classified by Maximum 
Severity (Safety Population) 
 
1.4.1.11 
2.4.1.11 
Subjects with solicited local 
Adverse Events by Symptom 
classified by Maximum Severity 
(Safety Population) 
 
1.4.1.12 
2.4.1.12 
Subjects with solicited systemic 
Adverse Events by Symptom 
classified by Maximum Severity 
(Safety Population) 
 
Unsolicited Adverse Events 
1.4.1.13 
2.4.1.13 
Subjects with unsolicited Adverse 
Events by PT (Safety Population) 
 
1.4.1.14 
2.4.1.14 
Subjects with unsolicited Adverse 
Events by SOC and PT (Safety 
Population) 
 
1.4.1.15 
2.4.1.15 
Subjects with related unsolicited 
Adverse Events by SOC and PT 
(Safety Population) 
 
1.4.1.16 
2.4.1.16 
Subjects with severe unsolicited 
Adverse Events by SOC and PT 
(Safety Population) 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 27 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
1.4.1.17 
2.4.1.17 
Subjects with serious unsolicited 
Adverse Events by SOC and PT 
(Safety Population) 
 
1.4.1.18 
2.4.1.18 
Subjects with medically attended 
unsolicited Adverse Events by 
SOC and PT (Safety Population) 
 
1.4.1.19 
2.4.1.19 
Subjects with related severe 
unsolicited Adverse Events by 
SOC and PT (Safety Population) 
 
1.4.1.20 
2.4.1.20 
Subjects with related serious 
unsolicited Adverse Events by 
SOC and PT (Safety Population) 
 
1.4.1.21 
2.4.1.21 
Subjects with related medically 
attended unsolicited Adverse 
Events by SOC and PT (Safety 
Population) 
 
1.4.1.22 
2.4.1.22 
Subjects with unsolicited Adverse 
Events leading to second 
Vaccination not administered 
(Safety Population) 
 
1.4.1.23 
2.4.1.23 
Subjects with unsolicited Adverse 
Events caused study 
discontinuation (Safety 
Population) 
 
1.4.1.24 
2.4.1.24 
Subjects with at least one 
unsolicited 
Adverse Event by Maximum 
Severity (Safety Population) 
 
1.4.1.25 
2.4.1.25 
Subjects with at least one 
medically attended unsolicited 
Adverse Event by Maximum 
Severity (Safety Population) 
 
1.4.1.26 
2.4.1.26 
Subjects with at least one 
unsolicited 
Adverse Event by Causality 
(Safety Population) 
 
1.4.1.27 
2.4.1.27 
Subjects with at least one 
medically attended unsolicited 
Adverse Event by Causality 
(Safety Population) 
 
1.4.1.28 
2.4.1.28 
Subjects with at least one serious 
unsolicited Adverse Event by 
Causality (Safety Population) 
 
NOTE: Adverse Event tables will also be provided for final analysis but including all AEs reported during the 
study. For the preliminary analysis the wording “up to Day 56” and for the final analysis “up to study end” will be 
added in the title of AE Table x.4.1.2-x.4.1.28. 
Laboratory Parameters 
1.4.1.29 
n/a 
Absolute Values for Hematology 
Parameters by Parameter and 
Visit (Safety Population) 
 
1.4.1.30 
n/a 
Absolute Values for Clinical 
Chemistry Parameters by 
Parameter and Visit (Safety 
Population) 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 28 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
1.4.1.31 
n/a 
Absolute Values for Coagulation 
Parameters by Parameter and 
Visit (Safety Population) 
 
1.4.1.32 
n/a 
Absolute Changes from Baseline 
for Hematology Parameters by 
Parameter and Visit (Safety 
Population) 
 
1.4.1.33 
n/a 
Absolute Changes from Baseline 
for Clinical Chemistry Parameters 
by Parameter and Visit (Safety 
Population) 
 
1.4.1.34 
n/a 
Absolute Changes from Baseline 
for Coagulation Parameters by 
Parameter and Visit (Safety 
Population) 
 
1.4.1.35 
n/a 
Subjects with Hematology 
Parameters Outside Normal 
Range by Parameter and Visit 
(Safety Population) 
 
1.4.1.36 
n/a 
Subjects with Clinical Chemistry 
Parameters Outside Normal 
Range by Parameter and Visit 
(Safety Population) 
 
1.4.1.37 
n/a 
Subjects with Coagulation 
Parameters Outside Normal 
Range by Parameter and Visit 
(Safety Population) 
 
1.4.1.38 
n/a 
Urinalysis Results by Parameter 
and Visit (Safety Population) 
 
1.4.1.39 
n/a 
Absolute Changes from Baseline 
for pH and Specific Gravity by 
Visit (Safety Population) 
 
Other Safety Parameters 
1.4.1.40 
n/a 
Systolic Blood Pressure [mmHg] 
by Visit (Safety Population) 
 
1.4.1.41 
n/a 
Diastolic Blood Pressure [mmHg] 
by Visit (Safety Population) 
 
1.4.1.42 
n/a 
Pulse Rate [beats/min] by Visit 
(Safety Population) 
 
1.4.1.43 
n/a 
Body Temperature [°C] by Visit 
(Safety Population) 
 
 
 
 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 29 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
 
List of Data Listings 
No. for prel.  
Analysis 
No. for FA 
Legend 
Content 
Overall Subject Information 
1.1.1 
2.1.1 
Analysis Population Details 
Subject number, Treatment, received IMP, 
Safety Population,  mITT Population, PP 
Population, Reason not in PP Population, 
Randomization performed, Reason 
randomization not performed, randomization 
date/time, randomization number 
1.1.2 
2.1.2 
Screening Failures with Reason 
Subject number, Date of informed consent, 
Reason for screening failure (Subject’s 
withdrawal of consent, In/Ex criterion not 
fulfilled / exclusion criterion fulfilled, Other), 
Specification other reason 
1.1.3 
2.1.3 
Violated Entry Criteria 
Subject number, Visit, Criterion ID not met, 
Criterion description 
1.1.4 
2.1.4 
Attended Visits with Dates 
(Safety Population) 
Subject number, Treatment, Visit, date, age, 
Reason for visit not performed, reason for 
outside time window, reason for unscheduled 
visit 
1.1.5 
2.1.5 
Protocol Deviations (Safety 
Population) 
Subject number, Treatment, PD Category, 
Classification, Reason for classification, PD 
Description 
1.1.6 
2.1.6 
Study Status (Safety Population) 
Subject number, Treatment, Visit attendance 
status, Primary reason early 
termination/treatment discontinuation, Other 
reason specification, AE Term, Specify 
individual stopping rules, AE term (Stopping 
rules), Primary cause of death, Death date, 
Date of study discontinuation 
Baseline Evaluation  
1.2.1 
n/a 
Demographic Information (Safety 
Population) 
Subject number, Treatment, Date of birth, Age 
[years], Gender, Childbearing potential, 
reason, specification other,  Race, 
Specification other race 
1.2.2 
n/a 
Full Physical Examination at Visit 
0 (Safety Population) 
Subject number, Treatment, Examination 
performed, Reason not performed, Exam Date 
1.2.3 
n/a 
Medical History (Safety 
Population) 
Subject number, Treatment, Condition, 
Preferred Term (MedDRA [Version]), SOC 
(MedDRA [Version]), Start date, End date, 
Ongoing at study end 
1.2.4 
2.2.4 
Prior Medication (Safety 
Population) 
Subject number, Treatment, Medication or 
therapy, ATC Level 2 (WHODD [Version]), ATC 
Level 3 (WHODD [Version]), Indication 
category, Indication, Dose, Dose Unit, Dose 
Unit other, Frequency, Frequency other, Route, 
Route other, Start Date, Ongoing at study end, 
End Date 
1.2.5 
2.2.5 
Concomitant Medication (Safety 
Population) 
Subject number, Treatment, Medication or 
therapy, ATC Level 2 (WHODD [Version]), ATC 
Level 3 (WHODD [Version]), Indication 
category, Indication, Dose, Dose Unit, Dose 
Unit other, Frequency, Frequency other, Route, 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 30 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
Route other, Start Date, Ongoing at study end, 
End Date 
1.2.6 
n/a 
Vaccination History (Safety 
Population) 
Subject number, Treatment, Visit, Any 
vaccination history within the last three years 
prior to screening, Any new vaccination history 
since screening, Date, Vaccination/Indication, 
ATC Level 2 (WHODD [Version]), ATC Level 3 
(WHODD [Version]), 
1.2.7 
n/a 
Concomitant Procedure (Safety 
Population) 
Subject number, Treatment, Procedure, 
Start/End date, Ongoing at study end, 
Indication category, Indication 
1.2.8 
n/a 
Virology 
Subject number, Treatment, Virology test, Test 
performed, Reason, Result, Date 
Immunogenicity Analysis 
n/a 
2.3.1 
Functional Antibodies 
Assessment (VNT) (mITT 
Population) 
Subject number, Treatment, 
Visit, Sample performed, Reason not 
performed, Sample ID, Date/Time of sample, 
Titer [IU/ml], Serconversion, positive/negative 
value 
n/a 
2.3.2 
Functional Antibodies 
Assessment (ELISA) m(ITT 
Population) 
Subject number, Treatment, 
Visit, Sample performed, Reason not 
performed, Sample ID, Date/Time of sample, 
Titer [IU/ml] , Serconversion, positive/negative 
value 
n/a 
2.3.3 
Measles Antibodies Assessment 
(mITT Population) 
Subject number, Treatment, 
Visit, Sample performed, Reason not 
performed, Sample ID, Date/Time of sample, 
Titer [IU/ml] , positive/negative value 
n/a 
2.3.4 
T-cell details (mITT Population) 
Subject number, Treatment, 
Visit, Sample performed, Reason not 
performed, Sample ID, Date/Time of sample, 
Titer [IU/ml] , positive/negative value 
Safety Analysis 
Extent of Exposure 
1.4.1 
2.4.1 
Extent of Exposure (Safety 
Population) 
Subject number, Treatment, Date first 
vaccination, Date second vaccination, 
Treatment duration [days], Date informed 
consent, Date of last attended visit, Study 
duration [days] 
1.4.2 
2.4.2 
Study Medication Details (Safety 
Population) 
Subject number, Treatment, Visit, Vaccination 
performed, Reason, Date/time 
Adverse Events 
1.4.3 
2.4.3 
Adverse Events Part 1 (Safety 
Population) 
Subject number, Treatment, AE Term, MedDRA 
PT, MedDRA SOC, AE type, Start date/time, 
End date/time, Medically attended, Severity, 
Serious AE, SAE Criteria, Relationship to Study 
Treatment 
1.4.4 
2.4.4 
Adverse Events Part 2 (Safety 
Population) 
Subject number, Treatment, AE Term, Action 
taken with Study Treatment, Other Action 
taken, specification other action taken, 
outcome, Ongoing at final examination 
1.4.5 
2.4.5 
Unsolicited Adverse Events Part 1 
(Safety Population) 
Subject number, Treatment, AE Term, MedDRA 
PT, MedDRA SOC, AE type, Start date/time, 
End date/time, Medically attended, Severity, 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 31 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
Serious AE, SAE Criteria, Relationship to Study 
Treatment 
1.4.6 
2.4.6 
Unsolicited Adverse Events Part 2 
(Safety Population) 
Subject number, Treatment, AE Term, Action 
taken with Study Treatment, Other Action 
taken, specification other action taken, 
outcome, Ongoing at final examination 
1.4.7 
2.4.7 
Related Unsolicited Adverse 
Events Part 1 (Safety Population) 
See Listing x.4.5 
1.4.8 
2.4.8 
Related Unsolicited Adverse 
Events Part 2 (Safety Population) 
See Listing x.4.6 
1.4.9 
2.4.9 
Severe Unsolicited Adverse 
Events Part 1 (Safety Population) 
See Listing x.4.5 
1.4.10 
2.4.10 
Severe Unsolicited Adverse 
Events Part 2 (Safety Population) 
See Listing x.4.6 
1.4.11 
2.4.11 
Serious Unsolicited Adverse 
Events Part 1 (Safety Population) 
See Listing x.4.5 
1.4.12 
2.4.12 
Serious Unsolicited Adverse 
Events Part 2 (Safety Population) 
See Listing x.4.6 
1.4.13 
2.4.13 
Medically attended unsolicited 
Adverse Events Part 1 (Safety 
Population) 
See Listing x.4.5 
1.4.14 
2.4.14 
Medically attended unsolicited 
Adverse Events Part 2 (Safety 
Population) 
See Listing x.4.6 
1.4.15 
2.4.15 
Unsolicited Adverse Events 
caused Study Discontinuation 
Part 1 (Safety Population) 
See Listing x.4.5 
1.4.16 
2.4.16 
Unsolicited Adverse Events 
caused Study Discontinuation 
Part 2 (Safety Population) 
See Listing x.4.6 
1.4.17 
2.4.17 
Unsolicited Adverse Events 
leading to second Vaccination 
not administered Part 1 (Safety 
Population) 
See Listing x.4.5 
1.4.18 
2.4.18 
Unsolicited Adverse Events 
leading to second Vaccination 
not administered Part 2 (Safety 
Population) 
See Listing x.4.6 
1.4.19 
2.4.19 
Solicited Adverse Events Part 1 
(Safety Population) 
Subject number, Treatment (Safety), Record 
ID, AE Term, Start date/time, End date/time, 
Medically attended, Severity, Serious AE, SAE 
Criteria, Causality 
1.4.20 
2.4.20 
Solicited Adverse Events Part 2 
(Safety Population) 
Subject number, Treatment (Safety), Record 
ID, AE Term, Action taken with Study 
Treatment, Other Action taken, Outcome, 
Ongoing at final examination, Caused study 
discontinuation 
1.4.21 
2.4.21 
Related Solicited Adverse Events 
Part 1 (Safety Population) 
See Listing x.4.19 
1.4.22 
2.4.22 
Related Solicited Adverse Events 
Part 2 (Safety Population) 
See Listing x.4.20 
1.4.23 
2.4.23 
Severe Solicited Adverse Events 
Part 1 (Safety Population) 
See Listing x.4.19 
1.4.24 
2.4.24 
Severe Solicited Adverse Events 
Part 2 (Safety Population) 
See Listing x.4.20 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 32 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
1.4.25 
2.4.25 
Serious Solicited Adverse Events 
Part 1 (Safety Population) 
See Listing x.4.19 
1.4.26 
2.4.26 
Serious Solicited Adverse Events 
Part 2 (Safety Population) 
See Listing x.4.20 
1.4.27 
2.4.27 
Medically attended Solicited 
Adverse Events Part 1 (Safety 
Population) 
See Listing x.4.19 
1.4.28 
2.4.28 
Medically attended Solicited 
Adverse Events Part 2 (Safety 
Population) 
See Listing x.4.20 
1.4.29 
2.4.29 
Solicited Adverse Events caused 
Study Discontinuation Part 1 
(Safety Population) 
See Listing x.4.19 
1.4.30 
2.4.30 
Solicited Adverse Events caused 
Study Discontinuation Part 2 
(Safety Population) 
See Listing x.4.20 
1.4.31 
2.4.31 
Solicited Adverse Events leading 
to second Vaccination not 
administered Part 1 (Safety 
Population) 
See Listing x.4.19 
1.4.32 
2.4.32 
Solicited Adverse Events leading 
to second Vaccination not 
administered Part 2 (Safety 
Population) 
See Listing x.4.20 
Laboratory Parameters 
1.4.33 
n/a 
Hematology Values outside 
Normal Range (Safety 
Population) 
Subject number, Treatment, Visit, Date/Time, 
Parameter, Value, Unit, Not done, Reason not 
done, Clinically significant, Likely cause, 
Change from baseline 
1.4.34 
n/a 
Clinical Chemistry Values outside 
Normal Range (Safety 
Population) 
Subject number, Treatment, Visit, Date/Time, 
Parameter, Value, Unit, Not done, Reason not 
done, Clinically significant, Likely cause, 
Change from baseline 
1.4.35 
n/a 
Coagulation Values outside 
Normal Range (Safety 
Population) 
Subject number, Treatment, Visit, Date/Time, 
Parameter, Value, Unit, Not done, Reason not 
done, Clinically significant, Likely cause, 
Change from baseline 
1.4.36 
n/a 
Abnormal and Positive Urinalysis 
Values (Safety Population) 
Subject number, Treatment, Visit, Date/Time, 
Parameter, Value, Unit, Not done, Reason not 
done, Clinically significant, Abnormal finding, 
Likely cause 
Other Safety Parameters 
1.4.37 
n/a 
Vital Signs (Safety Population) 
Subject number, Treatment, Visit, visit 
specifier, visit specifier, Vital signs collected, 
Reason not collected, Date/Time, Test, Result, 
Unit, Clinically significant, not done 
1.4.38 
n/a 
Symptom-directed Physical Exam 
(Safety Population) 
Subject number, Treatment, Exam performed, 
Reason not performed, Exam date/Time 
1.4.39 
n/a 
System Based Physical Exam 
(Safety Population) 
Subject number, Treatment, Exam performed, 
Reason not performed, other reason, Exam 
date/time 
1.4.40 
n/a 
Local Tolerability (Safety 
Population) 
Subject number, Treatment, visit, visit 
specifier, Assessment performed, Reason not 
performed, Exam Date/Time 
06G93N
07W9WK

SAP (MV-ZIKA-RSP-101, Final 3.0, 24-Sep-2020) 
Adapted from: 
 
Page 33 of 33 
STAT03_A Statistical Analysis Plan 
Version 6.0, Effective Date 16-Feb-2018 
Confidential and intellectual property of Assign DMB. No part of this document or its contents may be duplicated, referenced, published or 
otherwise disclosed in any form or by any means without prior written consent of Assign DMB. 
1.4.41 
n/a 
Pregnancy test (Safety 
Population) 
Subject number, Treatment, visit, Pregnancy 
test performed, Reason not performed, 
Date/time, Result 
 
8. 
REFERENCES 
Not applicable. 
06G93N
07W9WK