A Single Centre, Open Label, Pilot Phase Ib Study to Investigate Blood Stage Malaria Infection After Direct Venous Inoculation of Cryopreserved P. Falciparum (NF54 Strain) Sporozoites (PfSPZ-DVI) in Malaria naïve Healthy Adult Volunteers
Phase: PHASE1
Conditions: Malaria,Falciparum, Parasitemia, Protozoan Infections, Parasitic Disease
Sponsor: Medicines for Malaria Venture

Life Sciences, a division 
of SGS Belgium NV
REVISED CLINICAL STUDY PROTOCOL
CONFIDENTIALITY STATEMENT
The information in this document contains trade secrets and commercial information that are privileged 
or confidential and may not be disclosed unless such disclosure is required by applicable law or 
regulations. In any event, persons to whom the information is disclosed must be informed that the 
information is privileged or confidential and may not be further disclosed without
written authorization of MMV.
This study will be conducted in compliance with this protocol, 
the ICH Note for Guidance on Good Clinical Practice (CPMP/ICH/135/95) 
and with the applicable regulatory requirement(s).
A single centre, open label, pilot phase Ib study to investigate blood 
stage malaria infection after Direct Venous Inoculation of 
cryopreserved Plasmodium falciparum (NF54 strain) Sporozoites 
(PfSPZ-DVI) in malaria naïve healthy adult volunteers
Product
Riamet® (artemether-lumefantrine)
Protocol Number
MMV_PfSPZ-DVI Blood Stage_19_01
EudraCT Number
2019-004317-14
Clinical Phase
Ib
Clinical Indication
Acute Uncomplicated Plasmodium falciparum Malaria
in Adults
Issue Date (Version)
24-Jun-2020 (Final 3.0)
(Final Approved Protocol Amendment 1 dated 
27-Jan-2020 [Final 2.0])
Sponsor
Medicines for Malaria Venture (MMV)
Sponsor Representative
Farouk Chughlay, MD
Phone: +41 22 555 0355
Mobile: +41 79 123 4289
Fax: +41 22 555 0369

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SIGNATURES 
Signature of Sponsor Representative
Title:
A single centre, open label, pilot phase Ib study to investigate blood stage 
malaria infection after Direct Venous Inoculation of cryopreserved 
Plasmodium falciparum (NF54 strain) Sporozoites (PfSPZ-DVI) in 
malaria naïve healthy adult volunteers 
Name:
Farouk Chughlay, MD 
‘This Revised Clinical Study Protocol has been reviewed and approved by the Sponsor 
to ensure compliance with Good Clinical Practice.’ 
Signature: 
Date:
24 June 2020

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PROTOCOL HISTORY
Protocol Historya
Medicines for Malaria Venture (MMV) – MMV_PfSPZ-DVI Blood Stage_19_01
Document 
Issue Date
Amendment Type
Comments
Final Approved Protocol 
[Final 1.0]
17-Dec-2019
Not applicable
-
Amendment 1
[Final 2.0]
27-Jan-2020
Substantial
Revised Clinical Study Protocol 2.0
Amendment 2
[Final 3.0]
24-Jun-2020
Substantial
Revised Clinical Study Protocol 3.0
a This overview only lists general amendments to the protocol. Site- and country-specific amendments to 
the protocol are not included.

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TABLE OF CONTENTS
Title Page........................................................................................................................... 1
Signatures.......................................................................................................................... 2 
Protocol History................................................................................................................ 4 
Table of Contents.............................................................................................................. 5 
Protocol Amendment 2: Summary of Changes.............................................................. 9 
Protocol Amendment 1: Summary of Changes............................................................ 11 
Protocol Synopsis............................................................................................................ 15 
Time and Events Schedule............................................................................................. 29 
List of Abbreviations and Definitions of Terms .......................................................... 39 
Study Administrative Structure and Investigators ..................................................... 42 
1. 
Introduction ............................................................................................................ 44 
1.1 
Malaria Background Information....................................................................... 44 
1.2 
Malaria CHMI/VIS Models................................................................................ 44 
1.3 
Plasmodium falciparum Sporozoites Challenge Model (PfSPZ Challenge)...... 44 
1.4 
Antimalarial Therapy.......................................................................................... 46 
1.5 
Overall Rationale for the Study.......................................................................... 46 
1.6 
Risk Benefit Analysis......................................................................................... 46 
1.6.1 
Potential Risks...................................................................................... 46 
1.6.2 
Potential Benefits ................................................................................. 48 
1.6.3 
Conclusion............................................................................................ 48 
2. 
Objectives and Endpoints ...................................................................................... 49 
3. 
Study Design............................................................................................................ 53 
3.1 
Overview of Study Design ................................................................................. 53 
3.2 
Discussion of Study Design and Dose Selection................................................ 55 
3.3 
End of Study Definition...................................................................................... 60 
3.4 
Safety Review Committee (SRT)....................................................................... 60 
3.4.1 
Toxicity Criteria and Progression From Cohort 1 to Cohort 2 ............ 61 
3.4.2 
Final SRT Review at Study End .......................................................... 61 
4. 
Selection of Study Population................................................................................ 62 
4.1 
Inclusion Criteria................................................................................................ 62 
4.2 
Exclusion Criteria............................................................................................... 63 
4.3 
Lifestyle considerations...................................................................................... 66 
4.3.1 
Meals and Dietary Restrictions ............................................................ 66 
4.3.2 
Caffeine, Alcohol, and Tobacco........................................................... 66 
4.3.3 
Activity................................................................................................. 67 
4.3.4 
Other..................................................................................................... 67 
4.4 
Screen Failures ................................................................................................... 67 
5. 
Study Interventions ................................................................................................ 68 
5.1 
Study Interventions Administered...................................................................... 68 

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5.2 
Preparation/Handling/Storage/Accountability ................................................... 69 
5.2.1 
PfSPZ-DVI Challenge.......................................................................... 70 
5.2.2 
Registered Antimalarial Therapy ......................................................... 71 
5.3 
Randomisation and Blinding.............................................................................. 72 
5.4 
Dose Modification.............................................................................................. 72 
5.5 
Treatment Compliance ....................................................................................... 72 
6. 
Prior and Concomitant Therapy........................................................................... 73 
6.1 
Permitted Concomitant Therapies ...................................................................... 73 
6.2 
Prohibited Concomitant Therapies..................................................................... 73 
7. 
Assessments ............................................................................................................. 74 
7.1 
Timing of Assessments....................................................................................... 74 
7.1.1 
Screening Period .................................................................................. 74 
7.1.2 
Treatment Period.................................................................................. 75 
7.1.3 
Follow-up Period.................................................................................. 76 
7.1.4 
Unscheduled Visits............................................................................... 77 
7.2 
Pharmacodynamic Assessments......................................................................... 77 
7.2.1 
Parasitaemia ......................................................................................... 77 
7.3 
Safety Assessments ............................................................................................ 77 
7.3.1 
Adverse Events..................................................................................... 77 
7.3.2 
Malaria Clinical Score.......................................................................... 78 
7.3.3 
Clinical Laboratory Tests..................................................................... 79 
7.3.4 
Vital Signs............................................................................................ 80 
7.3.5 
Electrocardiogram ................................................................................ 80 
7.3.6 
Physical Examination........................................................................... 80 
7.3.7 
Beck Depression Inventory .................................................................. 81 
7.4 
Parasite Transcriptomics .................................................................................... 81 
7.5 
Treatment of Overdose....................................................................................... 81 
7.6 
Total Volume of Blood Sampling ...................................................................... 81 
7.7 
Appropriateness of Measurements ..................................................................... 81 
8. 
Participant Withdrawal from the Study............................................................... 82 
8.1 
Withdrawal of Participants From Study............................................................. 82 
8.2 
Lost to Follow-up ............................................................................................... 83 
9. 
Statistical Methods.................................................................................................. 84 
9.1 
Statistical Analysis ............................................................................................. 84 
9.1.1 
Initial Characteristics Data of the Participant Sample ......................... 84 
9.1.2 
Pharmacodynamic Data........................................................................ 85 
9.1.3 
Safety Data ........................................................................................... 86 
9.2 
Determination of Sample Size............................................................................ 88 
10. Adverse Event Reporting....................................................................................... 89 
10.1 
Definitions.......................................................................................................... 89 
10.2 
Intensity of Adverse Events................................................................................ 94 
10.3 
Causality Assessment......................................................................................... 94 
10.4 
Action Taken Regarding the Investigational products....................................... 95 
10.5 
Outcome ............................................................................................................. 95 
10.6 
Recording of Adverse Events............................................................................. 96 
10.7 
Reporting of Serious Adverse Events to PrimeVigilance Ltd............................ 96 

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10.8 
Pregnancy ........................................................................................................... 96 
10.9 
Reporting of Serious Adverse Events to Competent Authorities/Ethics 
Committees......................................................................................................... 97 
11. Ethical Aspects........................................................................................................ 98 
11.1 
Study-Specific Design Considerations............................................................... 98 
11.2 
Regulatory Ethics Compliance........................................................................... 98 
11.2.1 
Investigator Responsibilities ................................................................ 98 
11.2.2 
Independent Ethics Committee or Institutional Review Board 
(IEC/IRB)............................................................................................. 98 
11.2.3 
Informed Consent............................................................................... 100 
11.2.4 
Privacy of Personal Data.................................................................... 101 
12. Administrative Requirements ............................................................................. 102 
12.1 
Protocol Amendments ...................................................................................... 102 
12.2 
Participant Identification, Enrolment and Screening Logs............................... 102 
12.3 
Source Documentation ..................................................................................... 102 
12.4 
Case Report Form Completion......................................................................... 103 
12.5 
Monitoring........................................................................................................ 103 
12.6 
Data Management............................................................................................. 103 
12.7 
Data Quality Assurance.................................................................................... 104 
12.8 
On-Site Audits.................................................................................................. 104 
12.9 
Study Termination............................................................................................ 105 
12.10 
Record Retention.............................................................................................. 105 
12.11 
Use of Information and Publication ................................................................. 105 
12.12 
Registration of Clinical Studies and Disclosure of Results.............................. 107 
12.13 
Confidentiality.................................................................................................. 107 
13. References.............................................................................................................. 108 
Attachment 1: Clinical Score For Malaria................................................................. 111 
Attachment 2: Beck Depression Inventory................................................................. 112 
Attachment 3: Normal Ranges for Vital Signs and ECG ......................................... 115 
Attachment 4: Clinically Acceptable Ranges for Clinically Important Study 
Inclusion Laboratory Tests............................................................................ 116 

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List of Tables
Table 1: 
Objectives and Endpoints .............................................................................. 49 
Table 2: 
Description of Study Interventions................................................................ 69 
List of Figures
Figure 1: 
Schematic Overview of the Study for Cohort 1............................................. 55 
Figure 2: 
Schematic Overview of the Study for Cohort 2............................................. 55 

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PROTOCOL AMENDMENT 2: SUMMARY OF CHANGES
This amendment is considered to be substantial based on the criteria set forth in 
Article 10(a) of Directive 2001/20/EC of the European Parliament and the Council of the 
European Union.
Overall Reason for Amending the Protocol: This amendment was created to 
implement the Sponsor’s safety measures in the protocol with regard to the coronavirus 
disease 2019 pandemic. 
Changes are summarized below together with a rationale for each change.
1. Change in Exclusion Criteria
Rationale:
Per Sponsor’s decision, the study exclusion criteria have been updated to exclude any subject 
who tests positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2): since 
participants could experience flu-like symptoms following Direct Venous Inoculation of 
cryopreserved Plasmodium falciparum, for the safety of the participants, it is important to 
screen for SARS-CoV-2 infection prior to inoculation. Also before second confinement to the 
study site, participants should be tested for SARS-CoV-2 infection, and must be withdrawn 
from the study if the test result is positive.
Testing before first confinement to the study site (from the participants who already completed 
the other screening tests and are still eligible to be enrolled in the study) and before second 
confinement to the study site will be performed by taking a nasopharyngeal swab to detect 
presence of SARS-CoV-2 ribonucleic acid, using a real-time reverse transcription polymerase 
chain reaction (rRT-PCR) test.
Applicable 
Section(s)
Description of Changes
Synopsis
An additional exclusion criterion 24 was added to exclude any subject 
with SARS-CoV-2 infection. 
Time and Events 
Schedule
The schedule was updated to reflect the additional visits which can take 
place up to Day -2 (before the first confinement to the study site) and on 
Day 7-9 (before the second confinement), on which the nasopharyngeal 
swabs will be taken.
Section 4.2
An additional exclusion criterion 24 was added to exclude any subject 
with SARS-CoV-2 infection.
Section 7.1.1
The text on the screening of participants was updated to reflect the need 
for SARS-CoV-2 testing during screening.
Section 7.1.2
The text was updated to reflect the need for SARS-CoV-2 testing before 
the second confinement to the study site.
Section 8.1
The text was updated to reflect the need for participant withdrawal from 
the study if he or she tests positive for SARS-CoV-2 before confinement 
to the study site.

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2. Minor Corrections
Rationale:
Minor correction
Applicable 
Section(s)
Description of Changes
Synopsis
The text on the relationship of adverse events to malaria was updated.
The text on coagulation testing was updated to align with the text on the 
criterion for modified Hy’s law.
List of 
Abbreviations and 
Definitions of 
Terms
The list of abbreviations was updated.
Time and Events 
Schedule
The top rows of the flowchart were updated to clarify that Day -1 is not 
an actual part of the ‘Challenge, treatment and follow-up’ period.
The footnotes on coagulation testing were updated to align with the text 
on the criterion for modified Hy’s law.
The schedule was updated to include extra International normalized ratio
(INR) tests which will be performed post-baseline at the same time 
points as the post-baseline haematology and liver biochemistry tests.
Section 1.6.1
The text on total blood volume sampling was updated.
Section 5.1
The text on the relationship of adverse events to malaria was updated.
Section 7.1.1 and 
Section 7.1.2
The text on Day -1 was moved from Section 7.1.2 to Section 7.1.1 to 
clarify that Day -1 is not part of the treatment period.
Section 7.3.3
The text on coagulation testing was updated to align with the text on the 
criterion for modified Hy’s law.
Section 7.6
The text on total blood volume sampling was updated.
Section 8.1
The text on the relationship of adverse events to the PfSPZ-DVI 
Challenge agent and/or the antimalarial treatment was updated.
Section 10.1
Minor formatting corrections.
Section 10.3
The text on the relationship of adverse events to the PfSPZ-DVI 
Challenge agent and/or the antimalarial treatment was changed: this 
relationship is now classified as Related or Not related. The term 
‘suspected’ was removed from the category.
Section 10.7
The text was updated to clarify that Adverse Events of Special Interest 
must also be reported on the Serious Adverse Event Form.
Section 10.8
Induced abortion was removed from the list of abnormal pregnancy 
outcomes that are considered serious adverse events.

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PROTOCOL AMENDMENT 1: SUMMARY OF CHANGES
This amendment is considered to be substantial based on the criteria set forth in 
Article 10(a) of Directive 2001/20/EC of the European Parliament and the Council of the 
European Union
Overall Reason for Amending the Protocol: This amendment was created to make 
adaptations following review of the protocol by the Belgian Federal Agency for 
Medicines and Health Products (FAMHP).
Changes are summarized below together with a rationale for each change. 
1. Change in Exclusion Criteria
Rationale:
The study exclusion criteria have been updated based on the Agency’s recommendations to 
exclude any subject with hepatic disorders and in line with the Food and Drug Administration 
(FDA) Guidance pertaining to Drug-Induced Liver Injury (FDA, 20091).
1. The hepatitis blood test panel at study screening has been significantly expanded to ensure a 
comprehensive assessment of the most common causes of infectious hepatitis disorders in 
Phase 1 populations prior to study inclusion. Given that infection with hepatitis D virus only 
occurs in people who are already infected with the hepatitis B virus (HBV), testing for
hepatitis D virus will be performed exclusively in those participants who screen positive for 
HBV.
2. An exclusion criterion has been added to ensure that study participants with documented 
hepatic disorders other than common causes of hepatitis are not included (including but not 
limited to viral hepatitis, auto-immune hepatitis, non-alcoholic steatohepatitis [NASH], 
alpha-1-antitrypsin deficiency, alcoholic liver disease, primary biliary cholangitis [PBC], 
primary sclerosing cholangitis [PSC], hemochromatosis, Wilson disease or suspected 
hepatocellular carcinoma [HCC]).
Applicable 
Section(s)
Description of Changes
Synopsis
Exclusion criterion 6 was updated to expand the hepatitis blood test 
panel.
An additional exclusion criterion 7 was added to exclude any subject 
with hepatic disease. 
Serological testing in the safety assessments was updated to reflect the 
expanded hepatitis blood test panel.
Time and Events 
Schedule
Footnote related to serological testing was updated to reflect the 
expanded hepatitis blood test panel.
1 FDA Guidance for Industry Drug-Induced Liver Injury: Premarketing Clinical Evaluation, U.S. 
Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and 
Research (CDER) Center for Biologics Evaluation and Research (CBER), July 2009

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Applicable 
Section(s)
Description of Changes
Section 4.2
Exclusion criterion 6 was updated to expand the hepatitis blood test 
panel.
An additional exclusion criterion 7 was added to exclude any subject 
with hepatic disease.
Section 7.3.3
Serological testing was updated to reflect the expanded hepatitis blood 
test panel.
2. Change in Inclusion Criteria
Rationale:
The study inclusion criteria have been updated based on the Agency’s recommendation that 
liver transaminases at screening should not exceed the upper limit of normal (ULN) 
(>1x ULN, instead of >1.25x ULN). 
Applicable 
Section(s)
Description of Changes
Synopsis
Section 4.1
Inclusion criterion 4 has been updated to reflect that transaminases 
should not exceed >1x ULN.
3. Addition of Clinically Acceptable Ranges for Clinically Important Study 
Inclusion Laboratory Tests 
Rationale:
Based on the Agency’s recommendation, the Clinically Acceptable Ranges for Clinically 
Important Study Inclusion Laboratory Tests (including total and conjugated bilirubin) have
been added in Attachment 4 of the protocol to guide inclusion/exclusion of study participants.
Applicable 
Section(s)
Description of Changes
Section 4.1
Section 4.2
A reference to Attachment 4 was added.
Attachment 4
Clinically Acceptable Ranges for Clinically Important Study Inclusion 
Laboratory Tests have been added.

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4. Change in Composition of Safety Review Team (SRT)
Rationale:
Based on the Agency’s recommendation, and in line with the CHMP Guideline on data 
monitoring committees, an independent Early Phase and Clinical Pharmacology MD expert 
experienced in malaria volunteer infections studies was included as the chair of the SRT, in 
addition to the Medical Monitor, the Medical/Project Director, the Principal Investigator (PI)
and an independent infectious disease/malaria expert.
Applicable 
Section(s)
Description of Changes
Synopsis 
Section 3.4
Addition of an independent Early Phase and Clinical Pharmacology MD 
expert experienced in malaria volunteer infection studies as SRT chair to 
the corrected SRT composition.
5. Change in Toxicity Criteria
Rationale:
Based on the Agency’s recommendation, and in line with the European Medicines Agency 
(EMA) Guideline on strategies to identify and mitigate risks for first-in-human and early 
clinical trials with investigational medicinal products, the toxicity criteria taken into account to
progress from Cohort 1 to Cohort 2 were updated. The criterion on Riamet®-related AEs was 
changed from “two or more Riamet®-related severe (grade 3 or higher), same organ class AEs”
to “two or more Riamet®-related severe (grade 3 or higher) AEs, independent of within or not 
within the same system organ class”.
Applicable 
Section(s)
Description of Changes
Section 3.4.1
The toxicity criterion on Riamet®-related AEs was updated.

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6. Minor Corrections
Rationale:
Minor corrections
Applicable 
Section(s)
Description of Changes
Study 
Administrative 
Structure and 
Investigators
Section 12.5
Clinical Monitoring details were added.
Section 3.4
Section 3.4.1
The composition of the SRT was moved from Section 3.4.1 to the 
beginning of Section 3.4.
Synopsis
Section 3.4
Section 5.1
‘Malaria expert’ or ‘infectious diseases physician with expertise in 
malaria’ was replaced by ‘independent infectious disease/malaria 
expert’.
Synopsis
Time and Events 
Schedule
Section 7.3.2
Section 7.3.4
Temperature measurements were included into the vital signs part of the 
protocol because:
-
temperature measurement is required to assess the malaria clinical 
score - fever; but
-
the temperature also needs to be viewable as a data point for the 
medics.
Attachment 1
Timing of grading malaria clinical score was adapted.

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PROTOCOL SYNOPSIS
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PROTOCOL SYNOPSIS
Study Title
A single centre, open label, pilot phase Ib study to investigate blood stage 
malaria infection after Direct Venous Inoculation of cryopreserved 
Plasmodium falciparum (NF54 strain) Sporozoites (PfSPZ-DVI) in malaria 
naïve healthy adult volunteers
Product
Riamet®
(artemether-lumefantrine)
Clinical Phase
Ib
Protocol Number
MMV_PfSPZ-DVI Blood 
Stage_19_01
Indication
Acute Uncomplicated 
Plasmodium falciparum
Malaria in Adults
EudraCT Number
2019-004317-14
Sponsor
Medicines for Malaria Venture (MMV)
Sponsor Representative
Farouk Chughlay, MD
Clinical Centre
SGS Life Sciences, Clinical Pharmacology Unit, Lange 
Beeldekensstraat 267, 2060 Antwerpen, Belgium
Objectives and Endpoints:
OBJECTIVES
ENDPOINTS
Primary
x
To evaluate the safety and 
tolerability associated with 
blood-stage malaria 
infection in naïve healthy 
participants after 
inoculation of Plasmodium 
falciparum sporozoites 
through direct venous 
inoculation (PfSPZ-DVI 
Challenge).
x
The incidence and severity of observed (i.e., malaria signs and 
symptoms graded by the malaria clinical score) or self-reported 
adverse events (AEs) considered PfSPZ-DVI Challenge 
inoculum-related until end of study (EOS) visit.
x
The change in malaria clinical score from PfSPZ-DVI Challenge 
until parasite clearance (defined as a quantitative polymerase 
chain reaction [qPCR] value of 0 parasites per mL blood after 
initiating antimalarial therapy).
x
Changes from baseline in haematology, clinical chemistry and 
urinalysis parameters, vital signs and electrocardiogram (ECG) 
parameters.
x
To characterise key stages 
in the blood-stage parasite 
growth profile in malaria 
infection in naïve healthy 
participants after 
PfSPZ-DVI Challenge.
x
The parasite growth profiles of Plasmodium falciparum
(P. falciparum) NF54 after infection via injection of sporozoites 
by direct venous inoculation (DVI) will be characterised by:
o
time to first PCR positivity (i.e., time elapsed between 
PfSPZ-DVI Challenge and first qPCR outcome equal to or 
greater than 250 parasites per mL blood, with a maximum of 
28 days in the absence of parasitaemia);
o
parasitaemia at first PCR positivity;
o
WLPHWRSDUDVLWDHPLDRI parasites per mL blood 
(Cohorts 1 and 2);
o
parasitaePLDDWWKHWLPHSDUDVLWDHPLD parasites per mL 
blood (Cohorts 1 and 2);
o
WLPHWRSDUDVLWDHPLDRI parasites per mL blood 
(Cohort 2); 
o
SDUDVLWDHPLDDWWKHWLPHSDUDVLWDHPLD parasites per 
mL blood (Cohort 2);

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PROTOCOL SYNOPSIS
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o
time to first dose of treatment with artemether-lumefantrine 
(Riamet®) (Cohorts 1 and 2); and
o
parasitaemia at first dose of treatment with Riamet®
(Cohorts 1 and 2).
x
The number and proportion of participants with presence of 
SRVLWLYH3&5DQGSDUDVLWDHPLDRIRU parasites per 
mL blood between inoculation with PfSPZ-DVI Challenge and 
Day 28 (per cohort).
Secondary
x
To determine the safety and 
tolerability of Riamet® in 
blood-stage malaria 
infection in naïve healthy 
participants after 
PfSPZ-DVI Challenge.
x
The incidence, severity and relationship to Riamet® of observed 
or self-reported, treatment-emergent adverse events (TEAEs) until 
EOS visit.
x
To characterise the blood-
stage parasite profile in 
malaria infection in naïve 
healthy participants after 
PfSPZ-DVI Challenge.
x
The blood-stage parasite profile of P. falciparum NF54 after 
infection via injection of sporozoites by DVI will be characterised 
by: 
o
parasite growth rate expressed as the parasite multiplication 
rate (PMR) standardised to 48 h (PMR48) (Cohort 1, 
Cohort 2 and overall);
o
parasite growth rate expressed as the PMR per life cycle (if 
not 48 h) (PMRLC) (Cohort 1, Cohort 2 and overall); and
o
predicted time to reach parasitaemia threshold of first 
SRVLWLYH3&5SDUDVLWHVSHUP/EORRG
 parasites per mL blood for Cohorts 1 and 2.
x
To characterise the 
blood-stage parasite 
clearance profile of 
Riamet® in malaria 
infection in naïve healthy 
participants after 
PfSPZ-DVI Challenge.
x
The effect of Riamet® on clearance of P. falciparum blood-stage 
parasites will be evaluated using:
o
time to parasite clearance, i.e., time between drug 
administration and first PCR below the limit of 
quantification (Cohort 1, Cohort 2 and overall); 
o
log10 parasite reduction ratio per 48 h (log10PRR48), i.e., ratio 
of the parasite density at a specific time point to the parasite 
density 48 h later and expressed in log10 (Cohort 1, Cohort 2 
and overall); 
o
maximum log10PRR48 (log10PRR48,max) (Cohort 1, Cohort 2 
and overall); and
o
parasite clearance half-life (PC½), i.e., time taken for the 
parasite density to be reduced by 50% after the first dose 
administration of Riamet® (Cohort 1, Cohort 2 and overall).
Exploratory
x
To characterise the 
individual variability of 
blood-stage parasite profile.
x
The individual variability of blood-stage parasite profile will be 
characterised by:
o
the inter-individual variability of the parasite growth rate; 
and
o
predicted time to reach parasitaemia threshold of first 
SRVLWLYH3&5SDUDVLWHVSHUP/EORRG
parasites per mL blood including their 95% confidence 
interval (CI).

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PROTOCOL SYNOPSIS
17
Overview of Study Design:
This is a single-centre, open-label, Phase Ib study designed to assess if intravenous bolus injection of 
approximately 3200 P. falciparum (NF54 strain) sporozoites can be safely administered to achieve 
blood-stage parasitaemia with a kinetics/PCR profile that will allow for the future characterisation of 
antimalarial blood-stage activity of new chemical entities in a relatively small number of participants 
during early drug development.
Up to 16 healthy, malaria-naïve males and females, aged 18-55 years, will be enrolled in a maximum of 
2 cohorts (up to 8 participants per cohort; a participant may be enrolled in one cohort only). Enrolment 
into the cohorts will proceed sequentially, with two target levels of parasitaemia previously achieved in 
healthy participants enrolled in malaria Volunteer Infection Studies (VIS) at other study sites, i.e., 
5000 parasites/mL blood in Cohort 1 and 10000 parasites/mL blood in Cohort 2.
Each participant will be admitted to the clinical unit in the morning of Day -1 and inoculated with 
approximately 3200 P. falciparum sporozoites (NF54 strain) by DVI on Day 1. Participants will be 
discharged 2 h post inoculation on Day 1 and will be monitored daily via phone call from Day 2 until 
Day 6 to solicit any AEs. Participants will come to the clinical unit daily from Day 7 until Day 9 and 
together with the malaria clinical score, the presence of parasites will be assessed once daily by a specific 
qPCR targeting the varATS (the acidic terminal segment in Plasmodium falciparum var genes) multigenic 
IDPLO\WKLVWRDFFXUDWHO\GHVFULEHSDUDVLWHJURZWKHYHQLQFDVH3&5SRVLWLYLW\LHDT3&5RXWFRPH
parasites per mL blood, is confirmed this early (very low probability and with low densities). Participants 
will be confined to the clinical unit from Day 10 in the morning. qPCR will be performed and malaria 
clinical score assessed twice daily and participants will be administered registered antimalarial therapy, 
i.e., Riamet®, when the following criteria are met:
1.
&RKRUWSDUDVLtes/mL blood or earlier if a participant has a malaria clinical score >6 or at 
Investigator’s discretion.
2.
&RKRUWSDUDVLWHVP/EORRGRUHDUOLHULIDSDUWLFLSDQWKDVDPDODULDFOLQLFDOVFRUH!RU
at Investigator’s discretion.
The registered 3-day antimalarial therapy regimen will be further administered and monitored. qPCR 
assessments of parasitaemia will be carried out at multiple time points (2, 6, 8, 12, 16, 24, 36, 48 and 72 h) 
following initiation of Riamet® and malaria clinical score will be assessed twice daily during confinement 
in the clinical unit. Safety and tolerability will be monitored during the whole study duration, specific 
assessments will be done at periodic pre-specified time points from Day 10 and for at least 72 h after 
initiating antimalarial therapy, i.e., during confinement in the clinical unit (see below).
Of note, all participants must consent to receiving antimalarial therapy, i.e., the registered 3-day Riamet®
regimen approved for treatment of uncomplicated malaria. Even in the case of withdrawal from the study, 
all participants administered the PfSPZ-DVI Challenge are to receive antimalarial therapy as soon as 
possible, and to have all appropriate visits and assessments as required. If an intolerance or 
contraindication to Riamet® develops, Malarone® will be administered.
Upon parasite clearance (defined as a qPCR value of 0 parasites per mL blood after initiating antimalarial 
therapy) and at least 72 h after initiating antimalarial therapy (estimated to occur on or before Day 19 and 
on or before Day 22 for Cohort 1 and Cohort 2, respectively), and if clinically well, participants will be 
discharged from the clinical unit and will be followed up for safety assessments, clinical evaluation and 
malaria qPCR in the clinical unit at the EOS visit on Day 28. 
All participants who received antimalarial therapy will be asked non-leading questions to determine the 
occurrence of any AEs throughout the study and at the EOS visit.
All participants inoculated with PfSPZ-DVI Challenge will commence antimalarial therapy no later than 
Day 24 for both cohorts, regardless if they reach pre-defined cohort-specific PCR parasitaemia/malaria 
clinical score thresholds (i.e., 5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for 
&RKRUWDQGRUDPDODULDFOLQLFDOVFRUHIRU&RKRUWV 1 and 2). Participants who start antimalarial 
therapy on Day 24 will only be discharged from confinement at the end of the EOS visit on Day 28.
Antimalarial therapy may be initiated whenever deemed necessary by the Investigators, e.g., if there is a 
concern regarding the safety of a study participant. Therapy may be amended according to the treating 
physician if the participant does not respond to treatment or the condition worsens.

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PROTOCOL SYNOPSIS
18
A sentinel strategy will be employed in Cohorts 1 and 2. Each cohort will consist of 2 subgroups of 
participants, to be enrolled sequentially: Subgroup 1 will be composed of 2 participants; Subgroup 2 will 
be composed of 6 participants. Participants in Subgroup 2 will not be treated until the last participant in 
Subgroup 1 has completed antimalarial therapy and only upon decision of the Principal Investigator (PI), 
medical director and medical monitor after review of available safety and tolerability data (safety 
assessments, AEs and malaria clinical scores) and parasitaemia data, including clearance of parasitaemia, 
of Subgroup 1.
Review of available Cohort 1 safety/tolerability and parasitaemia data up to the last dose of Riamet® for 
all inoculated participants in Cohort 1 will be performed by a Safety Review Team (SRT) before 
enrolment of Cohort 2. For progression from Cohort 1 to Cohort 2, the SRT members will include at least 
an independent Early Phase and Clinical Pharmacology MD expert experienced in malaria volunteer 
infection studies (SRT chair), the Medical Monitor, the Medical/Project Director, the PI and an 
independent infectious disease/malaria expert (or their delegates).
Study Population:
Approximately 16 participants are planned to be enrolled sequentially in 2 cohorts of up to 8 participants 
per cohort. A participant may be enrolled in one cohort only.
Eligibility Criteria:
Inclusion Criteria:
1.
Informed Consent Form signed voluntarily before any study-related procedure is performed, 
indicating that the participant understands the purpose of and procedures required for the study 
and is willing to participate in the study, including administration of registered antimalarial 
therapy;
2.
Male or female, between 18 and 55 years old (extremes included) at screening;
3.
Body weight of at least 50 kg and a body mass index (BMI) of 19.0 to 30.0 kg/m2 (extremes 
included);
4.
Good general health without clinically relevant medical illness, physical exam findings including 
vital signs, and laboratory abnormalities (e.g., without liver transaminases >1x the upper limit of 
normal [ULN] and as defined in the protocol) as determined by the Investigator;
5.
Willing to adhere to the prohibitions and restrictions specified in this protocol, including 
willingness to stay confined to the inpatient unit for the required duration and willingness to 
avoid travelling outside of Benelux during the study period;
6.
Female participants should fulfil one of the following criteria:
a.
At least 1 year postmenopausal (amenorrhea >12 months and follicle stimulating hormone 
[FSH] >30 mIU/mL) prior to screening;
b.
Surgically sterile (bilateral oophorectomy, hysterectomy or bilateral salpingectomy);
c.
Will use contraceptives as outlined in inclusion criterion 7;
7.
Female participants of childbearing potential (excluding females with female partners) must 
agree to the use of a highly effective method of birth control from the screening visit until 
40 days after the EOS visit at Day 28 (covering a full menstrual cycle of 30 days starting after 
5 half-lives of last dose of Riamet®);
Note: Highly effective birth control methods include: combined (oestrogen- and 
progestogen-containing) oral/intravaginal/transdermal hormonal contraception associated with 
inhibition of ovulation, progestogen-only oral/injectable/implantable hormonal contraception 
associated with inhibition of ovulation, intrauterine device, intrauterine hormone-releasing 
system, bilateral tubal occlusion, vasectomised partner or sexual abstinence from heterosexual 
intercourse.
8.
Female participant has a negative pregnancy test at screening and upon admission in the clinical 
unit;
Note: Pregnancy testing will consist of VHUXPȕ-KXPDQFKRULRQLFJRQDGRWURSLQȕ-HCG) tests at 
VFUHHQLQJDQGDWWKH(26YLVLWDQGDXULQHȕ-HCG tests on Day -1, in all women.
9.
Different ways of being reachable 24/7 (e.g., by mobile phone, regular phone or electronic mail) 
during the whole study period.

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19
Exclusion Criteria:
1.
Nursing (lactating) women;
2.
Participation in any other clinical drug or vaccine study within 30 days (or 5 half-lives for drugs) 
preceding the day of PfSPZ-DVI Challenge (whichever is longer), or plans to participate in other 
investigational drug or vaccine research during the study period;
3.
Participants who took standard vaccinations within 3 months before the start of the study or are 
planning to take standard vaccinations during the study period up to 8 weeks after PfSPZ-DVI 
Challenge;
4.
Blood product donation to any blood bank during the 8 weeks (whole blood) or 4 weeks (plasma 
and platelets) prior to admission in the clinical unit on Day -1;
5.
Mean ECG outside normal range and deemed clinically relevant by the Investigator. Examples of 
clinically significant ECG abnormalities for this study include:
-
PR-interval >220 ms;
-
QRS-complex >120 ms;
-
Absolute QT greater than >500 ms;
-
QT interval corrected according to Bazett’s formula (QTcB) or QT interval corrected 
according to Fridericia’s formula (QTcF >450 ms for male participants, >470 ms for female 
participants;
-
Pathologic Q wave;
-
Significant ST-T wave changes;
-
Left or right ventricular hypertrophy;
-
Non-sinus rhythm except isolated premature atrial contractions and ventricular extrasystole 
<2 per 10 s ECG lead;
-
Incomplete left bundle branch block, or complete or intermittent right or left bundle branch 
block;
-
Second or third degree A-V heart block.
6.
Seropositive human immunodeficiency virus (HIV), hepatitis A immunoglobulin M (IgM)
antibody, hepatitis B virus (HBV) (hepatitis B surface antigen [HBsAg]), hepatitis C virus 
(HCV) (antibody), hepatitis D antibody, hepatitis E IgM antibody, cytomegalovirus (CMV) IgM 
antibody or Epstein Barr Virus (EBV) IgM antibody;
7.
Previous or current diagnosis of hepatitis including but not limited to viral hepatitis, 
auto-immune hepatitis, non-alcoholic steatohepatitis (NASH), alpha-1-antitrypsin deficiency, 
alcoholic liver disease, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC),
hemochromatosis, Wilson disease or suspected hepatocellular carcinoma (HCC).
8.
History or presence of diagnosed food or known drug allergies (including but not limited to 
allergy to any of the antimalarial medications to be used in the study), or history of anaphylaxis 
or other severe allergic reactions;
Note: Participants with seasonal allergies/hay fever, house dust mite or allergy to animals that are 
untreated and asymptomatic at the time of dosing can be enrolled in the study.
9.
History of convulsion or severe head trauma, excluding fever convulsion under 5 years of age;
Note: A medical history of a single febrile convulsion during childhood is not an exclusion 
criterion.
10. History of serious psychiatric condition that may affect participation in the study or preclude 
compliance with the protocol, including but not limited to past or present psychoses, disorders 
requiring lithium, a history of attempted or planned suicide, more than one previous episode of 
major depression, any previous single episode of major depression lasting for or requiring 
treatment for more than 6 months, or any episode of major depression during the 5 years 
preceding screening;
Note: The Beck Depression Inventory will be used as an objective tool for the assessment of 
depression at screening. In addition to the conditions listed above, participants with a score of 20 
or more on the Beck Depression Inventory and/or a response of 1, 2 or 3 for item 9 of this 
inventory (related to suicidal ideation) will not be eligible for participation. Participants with a 
Beck score of 17 to 19 may be enrolled at the discretion of the Investigator if they do not have a 
history of the psychiatric conditions mentioned in this criterion and their mental state is not 

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PROTOCOL SYNOPSIS
20
considered to pose additional risk to the health of the volunteer or to the execution of the study 
and interpretation of the data gathered.
11. A medical, occupational or family problem as a result of alcohol or illicit drug abuse during the 
past 12 months or current alcohol or illicit drug abuse or addiction (positive alcohol breath test or 
positive drug screen for amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine or 
opiates at screening or upon check-in at the clinical unit);
Note: Excessive use of alcohol is defined as an intake of >21 units per week for males and 
>14 units per week for females where one alcohol unit is defined as 10 mL or 8 g of pure alcohol. 
A single unit is equal to one 25-mL (single) measure of whisky (alcohol by volume [ABV] 40%), 
or a third of a pint of beer (190 mL; ABV 5-6%) or half a standard (175 mL) glass of wine (ABV 
12%).
12. Participants are non-smokers or ex-smokers for more than 90 days prior to screening, or smoke 
no more than 5 cigarettes per day. If users of nicotine products (i.e., spray, patch, e-cigarette, 
etc.), they should use the equivalent of no more than 5 cigarettes per day. Participants must agree 
to abstain from smoking while in the unit;
13. Use of any prescription drugs, herbal supplements (e.g., St John’s Wort) or over-the-counter 
medication within 7 days or 5 half-lives (whichever is longer) prior to the PfSPZ-DVI Challenge, 
or an anticipated requirement for the use of these during the course of the study;
Note: If necessary, the incidental use of non-steroidal anti-inflammatory drugs (NSAIDs) and 
paracetamol (2 g/day, 10 g/week) may be acceptable at the Investigator’s discretion and will be 
documented in the eSource system. The use of nutritional supplements during this time that are 
not believed to have the potential to affect participant safety nor the overall results of the study, 
may be permitted on a case-by-case basis by the Investigator.
14. Any surgical or medical condition possibly affecting drug absorption (e.g., cholecystectomy, 
gastrectomy, bowel disease), distribution, metabolism or excretion;
15. Personnel (e.g., Investigator, sub-investigator, research assistant, pharmacist, study coordinator 
or anyone mentioned in the delegation log) directly involved in the conduct of the study;
16. Any condition that in the opinion of the Investigator would jeopardise the safety or rights of a 
person participating in the study or would render the person unable to comply with the protocol;
17. Personal history of malaria;
18. Volunteer has travelled to or lived in a malaria-endemic area within 6 months prior to planned 
study enrolment;
19. Plans to travel to malaria-endemic region during the study period up to last follow-up visit;
20. Previous participation in any malaria vaccine or Controlled Human Malaria Infection (CHMI) 
study/VIS;
21. Falling in moderate or higher risk category for a fatal or non-fatal cardiovascular event within 
5 years (> 5%) determined by a validated risk estimation system, e.g., SCORE;
22. Use of systemic antibiotics with known antimalarial activity within 5 half-lives of PfSPZ-DVI 
Challenge (e.g., trimethoprim-sulfamethoxazole, doxycycline, tetracycline, clindamycin, 
erythromycin, fluoroquinolones or azithromycin) or an anticipated requirement for the use of 
these during the study period;
23. Receipt of blood or blood-derived products (including immunoglobulin) within 3 months prior to 
screening. Receipt of packed red blood cells (RBCs) given for an emergent indication in an 
otherwise healthy person, and not required as ongoing treatment is not exclusionary (for example 
packed RBCs emergently given during an elective surgery).
24. Participants who test positive for severe acute respiratory syndrome coronavirus 2 
(SARS-CoV-2).
Note: In case of an out-of-range clinical laboratory test (as defined in the protocol), vital sign or 
ECG value that will determine a participant’s eligibility, or in case of a positive drug screen, a 
retest or expert evaluation can be requested. Results of any retest must be available prior to 
inoculation. The result of the retest will be considered for participant eligibility at the 
Investigator’s discretion. Participants can be rescreened at the discretion of the Investigator.

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PROTOCOL SYNOPSIS
21
PfSPZ Challenge, Riamet® and Malarone®; Dose, Mode of Administration:
Manufacturing, packaging, and labelling of PfSPZ-DVI Challenge inoculum will be done under the 
responsibility of the biotechnology company Sanaria (USA).
The clinical unit will be responsible for acquiring the registered antimalarial drugs artemether 
lumefantrine (Riamet®) and atovaquone-proguanil (Malarone®):
-
Participants will be prescribed with Riamet® according to the pre-defined cohort-specific PCR 
parasitaemia/malaria clinical score thresholds (i.e., 5000 parasites/mL blood for Cohort 1 and 
SDUDVLWHVP/EORRGIRU&RKRUWDQGRUDPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQGWR
ensure parasite clearance prior to the end-of-study evaluation. In addition, participants will also 
be administered with Riamet®:
o
LIWKH\H[SHULHQFHLQRFXOXP-related serious adverse event (SAE), irrespective of severity; 
or
o
LIWKH\H[SHULHQFH&RPPRQ7HUPLQRORJ\&ULWHULDIRU$GYHUVH(YHQWV&7&$(
grade  (severe) AE deemed related to malaria and not self-resolved or relieved with 
concomitant medications; or 
o
if the PI or delegate considers it necessary for participant safety. In this situation, the PI or 
delegate will consult with the independent infectious disease/malaria expert (Prof. 
Jean-Pierre Van geertruyden). However, antimalarial medication may be administered prior 
to consultation if immediate treatment is deemed necessary for participant safety.
-
If an intolerance or contraindication to Riamet® develops, Malarone® will be administered.
All participants inoculated with PfSPZ-DVI Challenge will commence antimalarial therapy no later than 
Day 24 for both cohorts, regardless if they reach pre-defined cohort-specific PCR parasitaemia/malaria 
clinical score thresholds (i.e., 5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for 
&RKRUWDQGRUDPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQG
All participants must consent to receiving antimalarial therapy. Even in the case of withdrawal from the 
study, all participants administered the PfSPZ-DVI Challenge are to receive antimalarial therapy as soon 
as possible, and to have all appropriate visits and assessments as required.
The PfSPZ-DVI Challenge agent consists of a strain of P. falciparum NF54 sporozoites used for 
CHMI/VIS studies that is known to be sensitive to the registered antimalarial therapy treatments described 
above.

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01 24-Jun-2020 (Final 3.0)
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PROTOCOL SYNOPSIS
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Intervention Name
PfSPZ-DVI Challenge
artemether-lumefantrine 
(Riamet®)
atovaquone-proguanil 
(Malarone®); only to be 
used if an intolerance or 
contraindication to 
Riamet® develops
Intervention Type
inoculum
drug
Dose Formulation
cryovial
tablet
Unit Dose Strength(s)
15000 or 50000 aseptic, 
cryopreserved 
P. falciparum 
sporozoites
20 mg artemether and 
120 mg lumefantrine
250 mg atovaquone and 
100 mg proguanil 
hydrochloride
Dosage Level
3200 P. falciparum
sporozoites once
6 doses of 4 tablets over 
a period of 3 days at 
approximately 0, 8, 24, 
36, 48 and 60 h
4 tablets as a single 
daily dose for 
3 consecutive days
Route of 
Administration
intravenously by DVI
oral
Meals in Relation to 
Dosing
Not applicable
The first dose of antimalarial drug should be taken 
as soon as possible according to the pre-defined 
cohort-specific PCR parasitaemia/malaria clinical 
score thresholds (i.e., 5000 parasites/mL blood for 
Cohort 1 and 10000 parasites/mL blood for Cohort 
DQGRUDPDODULDFOLQLFDOVFRUHIRU&RKRUWV
and 2) or on reaching Day 24. Antimalarial drug 
administration should be immediately followed by 
a meal or drinks rich in fat (e.g., milk).
Use
challenge agent
antimalarial medication
Study/Treatment Duration:
It is estimated that each participant will be in the study for a maximum of 56 days, including a screening 
period of up to 28 days and a study period of 28 days, including PfSPZ-DVI Challenge on Day 1 and a 
follow-up period of 27 days afterwards.
Assessments:
Pharmacodynamics
Parasitaemia
A blood sample will be collected via direct venepuncture from each participant at the scheduled time points 
where parasitological assessments are scheduled. The assessment of malaria parasitology by qPCR will be 
as follows: varATS multigenic family targeted qPCR assay of parasite load will be performed in accordance 
with the centre of excellence for tropical medicine (ITM) standard operating procedure and the Laboratory 
Manual. Given the high sensitivity of qPCR, this method will be used to confirm parasite clearance after 
definitive antimalarial therapy for all participants. The results of the PCR at the ITM will be available in 
approximately 6 h from arrival of samples at the ITM. A participant will be considered cured following 
completion of the course of antimalarial therapy and after qPCR indicates 0 parasites per mL blood.
Safety
Adverse Events
Adverse events will be monitored continuously from informed consent until the last study-related activity. 
At regular intervals during the study, participants will be asked non-leading questions to determine the 
occurrence of any AEs. All AEs reported spontaneously during the course of the study will be recorded as 
well.
Close monitoring of expected signs and symptoms associated with malaria infection form part of the safety 
evaluation of the study and will be used as part of the decision criteria to administer registered antimalarial 

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PROTOCOL SYNOPSIS
23
therapy. These events will be classified as AEs, and may also be further sub-classified as inoculum-related 
AEs:
An inoculum-related AE is a sign or symptom associated with malaria infection, i.e., confirmed by a 
protocol-GHILQHG3&5SRVLWLYLW\GHILQHGIRUWKHSXUSRVHRIWKLVVWXG\DVDT3&5RXWFRPHSDUDVLWHV
per mL blood) at
-
the onset of the event; or
-
if the event started within 1 day of parasite clearance (defined as a qPCR value of 0 parasites per 
mL blood after initiating antimalarial therapy) or laboratory-defined PCR positivity (a qPCR 
outcome of 50-249 parasites per mL blood).
If the presence of P. falciparum malaria is confirmed (see above), then these AEs will be further 
sub-classified as inoculum-related AEs, following the guidelines on AE reporting, i.e., followed-up until 
resolution with the same causality assessment, and reported as such in the final clinical study report.
If the PCR results for P. falciparum are between 0 and <250 parasites per mL blood at the time of the onset 
of the event and the day before, usual AE/SAE reporting procedures and criteria will apply, but the event 
will not be classified as an inoculum-related AE.
Final classification of signs and symptoms as AE and inoculum-related AE, or as AE only, will occur after 
PCR results are available.
Malaria Clinical Score
The malaria clinical score, which assesses malaria signs and symptoms at the scheduled time points, will be 
part of the close monitoring for known/identified risks related to PfSPZ-DVI Challenge/malaria. The 
malaria clinical score has been widely used in the conduct of induced blood-stage malaria (IBSM)
CHMI/VIS studies in Australia and also implemented in the recently conducted PfSPZ-DVI 
Chemoprotection Challenge study with P218 (MMV_P218_17_01 study [EudraCT number: 
2018-003004-39]) at the SGS Clinical Pharmacology Unit in Antwerp, Belgium. This quantitative tool will
be used as part of the decision criteria to administer registered antimalarial therapy along with the level of 
parasitaemia.
The following 14 signs/symptoms frequently associated with malaria will be graded using a 4-point scale 
(absent: 0; mild: 1; moderate: 2; severe: 3) and summed to generate a total malaria clinical score (maximum 
score possible is 42): headache, myalgia (muscle ache), arthralgia (joint ache), fatigue/lethargy, malaise 
(general discomfort/uneasiness), chills/shivering/rigors, sweating/hot spells, anorexia, nausea, vomiting, 
abdominal discomfort, fever, tachycardia and hypotension. To determine severity of the 14 signs/symptoms 
we use the CTCAE grading scale grade 1 - 5. Mild (1) equates to CTCAE grade 1, Moderate (2) equates to 
CTCAE grade 2 and Severe (3) equates to CTCAE grade 3 or above. Individual scores for each symptom 
as well as the total score will be recorded.
For the grading of fever, the vital sign parameter body temperature (sublingual) will be assessed (see below).
Clinical Laboratory Tests
Blood samples will be collected by venepuncture or via indwelling cannula at the scheduled time points. 
Biochemistry and haematology testing will be performed on these samples, as well as viral serology testing 
(hepatitis A IgM antibody, HbsAg, anti-HCV antibody, hepatitis D antibody [only in subjects positive for 
HbsAg], hepatitis E IgM antibody, CMV IgM antibody, EBV IgM antibody and HIV) on the sample from 
VFUHHQLQJ,QDOOIHPDOHSDUWLFLSDQWVDOVRVHUXPȕ-HCG assessments at screening and at the EOS visit and 
DXULQHȕ-HCG assessment on Day-1 will be performed. FSH will be measured at screening in all women.
All blood samples for safety assessments should be taken in a fasted state (after overnight fast for at least 
8 h for unbiased glucose determination), except for the ones taken during antimalarial therapy for troponin T
PHDVXUHPHQWV DQG H[FHSW IRU WKH RQH RQ 'D\ RI ILUVW 3&5  SDUDVLWHVP/ LQ &RKRUW  RU
 parasites/mL in Cohort 2 and prior to initiating antimalarial therapy, where a non-fasted state is 
allowed.
Standard laboratory tests will be performed by ZNA Middelheim.

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The following biochemistry and haematology tests will be performed on the safety blood samples: 
-
Liver biochemistry: albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT),
alkaline phosphatase (ALP), gamma glutamylaminotransferase (GGT), total and direct bilirubin 
and total serum proteins;
-
Biochemistry other than liver: sodium, potassium, chloride, bicarbonate, urate, inorganic 
phosphate, creatinine, estimated glomerular filtration rate (eGFR), glucose, lactate 
dehydrogenase (LDH), blood urea nitrogen (BUN) and creatine phosphokinase (CPK); 
-
High sensitive troponin T and;
-
C-reactive protein (CRP);
-
Haematology: haemoglobin, haematocrit, RBC count, mean corpuscular haemoglobin (MCH), 
mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), white 
blood cell (WBC) count, platelet count, reticulocytes, neutrophils, eosinophils, basophils, 
lymphocytes and monocytes;
-
Coagulation: prothrombin time (PT) and activated partial thromboplastin time (aPTT) (at 
screening and on Day -1 only), and international normalized ratio (INR).
A midstream urine sample will be collected for urinalysis by dipstick for glucose, protein, nitrite, pH and 
occult blood at the scheduled time points for biochemistry other than liver. Microscopic examination for 
WBC, RBC and casts will be performed.
A urine drug screen (amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine and opiates) and 
an alcohol breath test will be performed at the scheduled time points.
The Investigator must review the laboratory report, document this review and record any change occurring 
during the study he/she considers to be clinically relevant in the eSource system. Laboratory values outside 
the normal range will be flagged and their clinical relevance will be assessed by the Investigator.
Vital Signs
Vital sign parameters will be assessed after at least 10 min in supine position at the scheduled time points. 
The vital sign parameters that will be assessed are supine systolic and diastolic blood pressure (SBP and 
DBP, respectively) and pulse rate. Body temperature (sublingual) will also be assessed, and this whenever 
the malaria clinical score will be evaluated (see above).Orthostatic changes to BP and pulse rate will also 
be assessed at screening: participants will be requested to stand after completion of the supine measurements 
and blood pressure and pulse rate will be recorded after at least 2 min in the standing position.
These parameters will be measured using a completely automated device consisting of an inflatable cuff 
and an oscillatory detection system. All values will be registered on a built-in recorder so that measurements 
are observer-independent.
Any change from baseline in vital sign values occurring during the study that is considered to be clinically 
relevant by the Investigator should be recorded in the eSource system.
Electrocardiogram
Twelve-lead ECG recordings will be recorded in triplicate after at least 10 min in supine position at the 
scheduled time points. 
Paper speed will be 25 mm/s, so that the different ECG intervals can be measured manually.
The interpretations of the ECGs will be performed by the Investigator or his/her designee at the clinical 
unit. Any change from baseline ECG occurring during the study that is considered to be clinically relevant 
by the Investigator should be recorded in the eSource system.
Physical Examination
Physical examination will be performed at the scheduled time points.
Height is to be measured barefoot and at screening only. Body weight is to be measured at the scheduled 
time points. To obtain the actual body weight, participants must be weighed lightly clothed. 
Any change in physical examination occurring during the study that is considered to be clinically relevant 
by the Investigator should be recorded in the eSource system.
Full physical examination will be conducted at screening, on Day -1, within ±24 h after first PCR 
 parasites/mL blood and at the EOS visit on Day 28. Targeted, symptom-driven physical examination 

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01 24-Jun-2020 (Final 3.0)
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25
will be performed in case malaria clinical score >6 on Day 10 (upon confinement to the clinical unit), within 
KDIWHUILUVW3&5&RKRUWRU&RKRUWSDUDVLWHVP/EORRGDQGDWSUH-discharge from 
clinical unit; and will be focused on changes since the previous examination, but will always include at 
least: chest/respiratory, general appearance, heart/cardiovascular, abdomen, neurological and skin/mucous 
membranes examination.
Beck Depression Inventory
The Beck Depression Inventory is performed at screening only.
The questionnaire is scored by the participant. The inventory completed by the participant will be 
reviewed/checked for completeness and the total score calculated by the study personnel.
Parasite Transcriptomics
Different environmental conditions that occur during a human infection (such as the presence of drugs) 
can select for parasites with different expression patterns. We aim to evaluate the effect of in-host factors 
on parasite transcriptional/epigenetic responses during experimental infections. 
To this aim, blood samples will be collected via direct venepuncture at the scheduled time points. Sample 
aliquots will be stored in Trizol for RNA preservation and cryopreserved for parasite culture in case 
higher parasite densities are needed to increase starting material for transcriptomic analysis of parasite 
isolates.
Protocols for transcriptomic analysis of parasite isolates have been validated and are currently in use at the 
Malariology Unit at the ITM.
Statistical Methods:
All statistical analysis will be performed by SGS Life Sciences, under the supervision and responsibility 
of the Sponsor, using SAS® (SAS Institute Inc., Cary, NC, USA; version 9.4 or higher) software for 
statistical computations. 
All statistical methods shall be detailed in a Statistical Analysis Plan (SAP) that will be finalised before 
database lock.
The final analysis will be performed once all participants have completed the follow-up visit or have 
discontinued earlier. 
Data from all participants enrolled and inoculated with PfSPZ-DVI Challenge will be included in the data 
analysis. The following analysis populations will be defined:
-
Inoculation Set: all participants who were inoculated with PfSPZ-DVI Challenge;
-
Pharmacodynamic (PD) Analysis Set: all inoculated participants with at least one available PD 
data who received all Riamet® doses and who experienced no protocol deviations with relevant 
impact on PD data;
-
Safety Analysis Set: all inoculated participants who received any treatment.
Unless specified otherwise, the Inoculation Set will be used for analysis of demographics, the PD analysis 
set will be used for pharmacodynamic statistical analysis and the Safety Analysis Set will be used for 
safety/tolerability analysis.
Sample Size
Up to 16 participants will be enrolled in 2 cohorts of 8 participants per cohort. In agreement with the 
Sponsor, additional participants may be recruited in each cohort, to replace discontinuations for non-safety 
reasons and achieve the required cohort size.
This is an exploratory study focusing on the methodology of malaria inoculation in healthy participants, 
thus no formal sample size calculation is performed.
Initial Characteristics Data of the Participant Sample
For all participants who receive PfSPZ-DVI Challenge, descriptive statistics will be provided for 
demographic (e.g., age, height, weight, BMI, race, gender) and other initial participant characteristics 
(alcohol and drug screening tests, pregnancy test, orthostatic changes to blood pressure and pulse rate, 
serology, medical and social history, concomitant diseases, Beck Depression Inventory). Standard 
descriptive statistics for continuous variables are the number of participants (N), mean, standard deviation 
(SD), median, minimum and maximum values. The standard descriptive statistics for categorical variables 
are the number of participants in the category and the proportion expressed as a percentage.
Prior and concomitant medications will be coded using the WHO_DRUG Dictionary.

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01 24-Jun-2020 (Final 3.0)
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26
Pharmacodynamic Data
Parasitaemia
Primary endpoints:
To characterise key stages in the parasite growth:
-
first PCR positivity;
-
threshold of 5000 parasites/mL for Cohort 1 and threshold of 10000 parasites/mL for Cohort 2;
and
-
parasitaemia levels at drug administration.
This will be done by calculating:
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of time to first PCR positivity. In the 
absence of positive PCR, the duration will be set to a maximum of 28 days;
-
Descriptive statistics (N, geometric mean, SD, 95% CI, range) of parasitaemia at first PCR 
positivity;
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of time to parasitaemia of 
 parasites per mL blood (Cohorts 1 and 2);
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of parasitaemia at the time 
SDUDVLWDHPLDSDUDVLWHVSHUP/EORRG&RKRUWVDQG
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of time to parasitaemia of 
 parasites per mL blood (Cohort 2); 
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of parasitaemia at the time 
SDUDVLWDHPLDSDUDVLWHVSHUP/EORRG&RKRUW
-
Descriptive statistics per cohort (geometric mean, SD, 95% CI, range) of time to first dose of 
treatment with Riamet® (Cohorts 1 and 2); and
-
Descriptive statistics per cohort (geometric mean, SD, 95% CI, range) of parasitaemia at first 
dose of treatment with Riamet® (Cohorts 1 and 2).
7KHQXPEHUDQGSURSRUWLRQRISDUWLFLSDQWVZLWKSUHVHQFHRISRVLWLYH3&5DQGSDUDVLWDHPLDRI or 
SDUDVLWHVSHUP/EORRGEHWZHHQLQRFXODWLRQZLWKPfSPZ-DVI Challenge and Day 28 will be 
summarised per cohort. Corresponding two-sided 90% Exact Clopper-Pearson confidence limits will be 
presented as well.
Secondary endpoints:
To characterise the blood-stage parasite profile, a model will be fitted to the measured parasitaemia data 
prior to Riamet® administration to be able to predict the growth of parasites as a function of time. This 
model may be a log-linear model or, if the parasitaemia profile shows a cyclic behaviour due to 
synchronicity of the parasites and their sequestration during the late stages of their lifecycle, it may 
include a sinus function. The latter will allow the estimation of the period of the observed waves in
addition to the parasite growth. The following parameters will be calculated from the individual fits of the 
data:
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) of parasite 
growth rate expressed as the PMR48;
-
If cycles are observed and if their estimated period is not 48 h, descriptive statistics per cohort 
and overall (geometric mean, SD, 95% CI, range) of parasite growth rate expressed as the 
PMRLC; and
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of predicted time to reach 
SDUDVLWDHPLDWKUHVKROGRIILUVWSRVLWLYH3&5SDUDVLWHVSHUP/EORRGSDUDVLWHVSHU
mL blood for Cohorts 1 and 2.
To characterise the blood-stage parasite clearance profile of Riamet® in malaria infection in naïve healthy 
participants after PfSPZ-DVI Challenge; by calculating:
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) of time to 
parasite clearance, i.e., time between drug administration and first PCR below the limit of 
quantification;
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) of log10PRR48;
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) of 
log10PRR48,max; and
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI range) of PC½.

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01 24-Jun-2020 (Final 3.0)
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27
The parasite reduction ratios (PRRs) and the clearance half-life will be estimated from a log-linear model 
that will be fitted to the parasitaemia data observed from the administration of the treatment onwards. The 
PRR48, PRR48,max and PC½ will be calculated from the individual parameters and summarised by cohort.
Exploratory endpoints:
To characterise the individual variability of blood-stage parasite profile: 
-
Obtain the inter-individual variability of the parasite growth rate; and
-
5th, 50th and 95th percentiles of predicted time to reach parasitaemia threshold of first positive 
3&5SDUDVLWHVSHUP/EORRGSDUDVLWHVSHUP/EORRGLQFOXGLQJWKHLU&,
Both these estimates will be based on the mixed model on parasite growth data described above.
Safety
Safety parameters will be tabulated and analysed descriptively.
Adverse Events
The original terms entered in the eSource system by Investigators to identify AEs will be fully described 
and coded according to the Medical Dictionary for Regulatory Activities (MedDRA).
The reported AEs will be allocated to phases based on their start date. All AEs will be listed. All AEs with 
onset after PfSPZ-DVI Challenge will be summarised by cohort. Summaries will be made per MedDRA 
primary system organ class, MedDRA preferred term, severity, with the number and percentage of 
participants and the number of events. Similar summaries will be prepared for AEs considered to be 
related to Riamet® and AEs considered to be related to the PfSPZ-DVI Challenge, for serious AEs and 
AEs of special interest.
Special attention will be paid to those participants who died, discontinued the investigational products due 
to an AE or experienced a severe or serious AE. Summaries, listings and narratives may be provided, as 
appropriate.
Malaria Clinical Score 
For the malaria clinical score that will be administered by the PI or his/her trained delegate, actual values 
and changes from baseline will be evaluated by means of descriptive statistics. Additionally, expected 
signs and symptoms will be summarised by score.
Clinical Laboratory Tests
Each continuous biochemistry and haematology laboratory test will be evaluated by means of descriptive 
statistics on the actual values, at each assessment time point and by cohort. Changes from baseline will 
also be summarised using descriptive statistics by assessment time point and by cohort.
Relative changes in clinical laboratory test values compared to values at baseline will be evaluated in 
accordance with the normal ranges of the clinical laboratory (below, within or above normal range). The 
percentage of participants with clinical laboratory test abnormalities will be summarised by cohort.
The number and percentage of participants with liver enzyme elevations after inoculation with the 
PfSPZ-DVI Challenge as defined below will be summarised:
-
ALT or AST >3 x ULN;
-
ALT or AST >5 x ULN;
-
ALT or AST >8 x ULN;
-
ALT or AST >3 x ULN and bilirubin >2 x ULN at the same time point, together with a 
conjugated bilirubin fraction> 35% (Potential Hy’s law cases).

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01 24-Jun-2020 (Final 3.0)
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28
A listing of participants with any clinical laboratory test result outside the reference ranges will be 
provided.
Vital Signs
Vital signs parameters will be assessed after at least 10 min in supine position at the scheduled time 
points. Pulse rate, SBP and DBP will be evaluated by means of descriptive statistics (actual values and 
changes from baseline). 
The percentage of participants with vital signs abnormalities will be summarised by cohort in a 
cross-tabulation of post-baseline versus baseline abnormalities to the normal ranges.
Electrocardiogram
Twelve-lead ECG recordings will be performed in triplicate after participants remained in a supine 
position for at least 10 min. 
All ECG data automatically measured by ECG devices (PR, QRS, QT, QTcB, QTcF and HR) and overall 
ECG evaluation will be listed. The ECG data, along with changes from baseline will be summarised by 
means of descriptive statistics at each assessment time point and by cohort. 
The percentage of participants with ECG abnormalities will be summarised by cohort in a cross-tabulation 
of post-baseline versus baseline abnormalities to the normal ranges. This cross-tabulation will include 
categorical assessment on actual values and changes from baseline of QTcB and QTcF prolongation.
Physical Examination
Abnormal findings in physical examination will be listed.

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01
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TIME AND EVENTS SCHEDULE
Cohort 1
Assessments
Screening Visit
Day -28 to Day -2
Challenge, treatment and follow-up
EOS Visit
Day 28v
Study Day/Event
Day -1a
Day 1
Day 2-6
Day 7-9
Day 10: Start 
of 
Confinement 
in Clinical 
Unit
Day of First PCR 
 Parasites/mL Until 
Day of First PCR 
 Parasites/mL
Day of First PCR 
 Parasites/mL Until 
 h After Initiating 
Antimalarial Therapy
Day of Discharge From 
Confinement: Day of 
Parasite Clearance 
(0 Parasites/mL) AND 
K$IWHU,QLWLDWLQJ
Antimalarial Therapy
Time point (h) in 
relation to 
PfSPZ-DVI 
Challenge
0b
2
Estimated On or Before 
Day 19
Ambulatory visit
X
X
X
Ambulatory visit 
(SARS-CoV-2 test)
X
X
Confinement in 
clinical unitc
X
X
X
X
X
(X)
Discharge from 
clinical unit
X
X
(X)
Telephonic 
Monitoring for AEs
X
Eligibility criteria
X
X
Informed consentd
X
Demographics
X
Medical and social 
history
X
Beck Depression 
Inventory
X
Alcohol & drug 
screene
X
X
Height & weightf
X
X
X
Physical 
examinationg
X
X
X
X
X
X
X
Vital signsh
X
X
X
X
X
X
X
X
X
12-lead ECGi
X
X
Viral serologyj
X
Pregnancy testk
X
X
X

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01
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30
Assessments
Screening Visit
Day -28 to Day -2
Challenge, treatment and follow-up
EOS Visit
Day 28v
Study Day/Event
Day -1a
Day 1
Day 2-6
Day 7-9
Day 10: Start 
of 
Confinement 
in Clinical 
Unit
Day of First PCR 
 Parasites/mL Until 
Day of First PCR 
 Parasites/mL
Day of First PCR 
 Parasites/mL Until 
 h After Initiating 
Antimalarial Therapy
Day of Discharge From 
Confinement: Day of 
Parasite Clearance 
(0 Parasites/mL) AND 
K$IWHU,QLWLDWLQJ
Antimalarial Therapy
Time point (h) in 
relation to 
PfSPZ-DVI 
Challenge
0b
2
Estimated On or Before 
Day 19
DVI of PfSPZ 
Challenge
X
Riamet®l
Xl
Malaria Clinical 
Scorem
X
X
X
X
X
X
X
Haematology &
liver biochemistryn
X
X
X
X
X
X
X
X
Biochemistry other 
than livero
X
X
X
X
X
CRP analysisp
X
X
X
X
X
X
X
Coagulation 
parametersq
X
X
X
X
X
X
X
X
rRT-PCR for 
SARS-CoV-2
Xr
Xs
qPCR for parasitest
X
X
X
X
X
X
X
Parasite 
Transcriptomicsu
X
X
X
X
X
X
Previous 
medications
X
Concomitant 
medications
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X
AEs
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X
SAE reporting
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X
ȕ-HCG: ȕ-human chorionic gonadotropin, AE: adverse event, aPTT: activated partial thromboplastin time, CMV: cytomegalovirus, CRP: C-reactive protein, DVI: direct venous 
inoculation, EBV: Epstein Barr Virus, ECG: electrocardiogram, EOS: end of study, FSH: follicle stimulating hormone, HbsAg: hepatitis B surface antigen , HCV: hepatitis C virus,
HIV: human immunodeficiency virus, IgM: immunoglobulin M, INR: international normalized ratio, PfSPZ: Plasmodium falciparum sporozoites, PT: prothrombin time,
qPCR: quantitative polymerase chain reaction, rRT-PCR: real-time reverse transcription polymerase chain reaction, SAE: serious adverse event, SARS-CoV-2: severe acute respiratory 
syndrome coronavirus 2

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01
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a. Assessments begin as of participant confinement in the clinical unit and must be completed and safety assessment outcomes must be available before PfSPZ-DVI Challenge on 
Day 1.
b. On Day 1, assessments begin as of 3 h before challenge (except for urinalysis assessments, morning void allowed). The assessments indicated must be completed and safety 
assessment outcomes must be available before PfSPZ-DVI Challenge inoculation.
c. Confinement to the clinical unit will occur as follows:
x
Participants will be admitted to the clinical unit in the morning of Day -1 and discharged from confinement 2 h post PfSPZ-DVI Challenge on Day 1.
x
Participants will be admitted to the clinical unit in the morning of Day 10 and discharged on Day of discharge from confinement (Day of parasite clearance $1'KDIWHU
initiating antimalarial therapy), estimated to occur on or before Day 19. Of note, participants who commence antimalarial therapy on Day 24 (see l) will only be discharged from 
confinement at the end of the EOS visit on Day 28.
d. No study-related procedure is to be performed before voluntarily signing of the informed consent form.
e. Alcohol breath test and urine dipstick screening for drug abuse.
f. Height to be measured at screening only; body weight to be measured at screening, at admission on Day -1 and at the EOS visit.
g. Physical examination will be conducted:
x
once at screening and once on Day -1;
x
once on Day 10 (upon admission to clinical unit for confinement); 
x
once within 24 h after ILUVW3&5SDUDVLWHVP/
x
once within 24 h after first 3&5SDUDVLWHVP/
x
once at pre-discharge from clinical unit on Day of discharge from confinement (Day of parasite FOHDUDQFH$1'KDIWHULQLWLDWLQJDQWLPDODULDOWKHUDS\); and
x
once at the EOS.
Full physical examination will be conducted at screening, on Day -1, within ±24 h after ILUVW3&5SDUDVLWHVP/ and at the EOS visit on Day 28. Targeted (symptom-driven) 
physical examination will be performed on the other time points in case malaria clinical score >6.
h. Vital signs (blood pressure and pulse) will be measured after remaining in a supine position for at least 10 min. All measurements will be performed as follows:
x
once at screening and once on Day -1;
x
on Day 1 (pre and post PfSPZ-DVI Challenge);
x
twice daily from Day 10 (admission to clinical unit for confinement) onwards until Day of discharge from confinement (Day of SDUDVLWHFOHDUDQFH$1' h after initiating 
antimalarial therapy); and
x
once at the EOS.
Body temperature (sublingual) will also be assessed, and this whenever the malaria clinical score will be evaluated (see m).
At screening, orthostatic changes to blood pressure and pulse rate will also be assessed: participants will be requested to stand after completion of the supine measurements and 
blood pressure and pulse rate will be recorded after at least 2 min in the standing position.
i. 12-lead ECGs recordings will be performed in triplicate after participants have remained in a supine position for at least 10 min. All recordings will be performed as follows:
x
once at screening; and
x
once on Days 2 and 3 of antimalarial therapy.
j. Serological testing for HIV, hepatitis A IgM antibody, HbsAg, anti-HCV antibody, hepatitis D antibody (only in subjects positive for HbsAg), hepatitis E IgM antibody, CMV IgM 
antibody and EBV IgM antibody, to determine eligibility for the study.
k. 3UHJQDQF\WHVWLQJFRQVLVWVRIVHUXPȕ-HCG assessments at screening and at the EOS visit and a XULQHȕ-HCG assessment on Day-1.
l. Every participant will be prescribed a 3-day oral course of Riamet® (20 mg artemether and 120 mg lumefantrine tablets) (see Section 5.1). The first dose should be taken as soon as 
possible when a PCR threshold of 5000 parasites/mL is achieved or earlier if a participant has a malaria clinical score >6 or at Investigator’s discretion. Of note, all participants 
inoculated with PfSPZ-DVI Challenge will commence antimalarial therapy no later than Day 24, regardless if they reach the pre-defined cohort-specific PCR parasitaemia/malaria 
clinical score thresholds (i.e., 5000 parasites/mL blood and/or DPDODULDFOLQLFDOVFRUH (see Section 4.3).
If an intolerance or contraindication to Riamet® develops since inclusion, Malarone® will be administered (see Section 5.1).

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01
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32
m.Malaria clinical score for malaria signs and symptoms and the vital sign parameter body temperature (sublingual) (see h) are assessed:
x
on Day 1 (pre-PfSPZ-DVI Challenge);
x
once daily during ambulatory visits on Day 7, 8 and 9;
x
twice daily from Day 10 (admission to clinical unit for confinement) onwards until Day of discharge from confinement (Day of parasite clearance AND KDIWHULQLWLDWLQJ
antimalarial therapy); and
x
once at the EOS.
n. Haematology and liver biochemistry laboratory tests will be performed after overnight fast for at least 8 h (see Section 4.3):
x
once at screening and once on Day -1;
x
once during ambulatory visit on Day 7;
x
once on Day 10 (upon admission to clinical unit for confinement);
x
once within 24 h after ILUVW3&5SDUDVLWHVP/;
x
once on Day of first 3&5SDUDVLWHVP/$1'SULRUWRLQLWLDWLQJDQWLPDODULDOWKHUDS\ (non-fasted state allowed);
x
once at pre-discharge from clinical unit on Day of discharge from confinement (Day of parasite clearance $1'KDIWHULQLWLDWLQJDQWLPDODULDOWKHUDS\); and
x
once at the EOS.
Troponin T will be measured at screening (fasted state), at baseline prior to inoculation on Day -1 (fasted state), and on Days 1, 2 and 3 of antimalarial therapy (non-fasted state).
FSH will be measured at screening in all women.
o. Biochemistry laboratory tests other than liver (including urinalysis; morning void allowed) will be performed after overnight fast for at least 8 h (see Section 4.3):
x
once at screening and once on Day -1;
x
once on Day 10 (upon admission to clinical unit for confinement);
x
RQFHRQ'D\RIILUVW3&5SDUDVLWHVP/$1'SULRUWRLQLWLDWLQJDQWLPDODULDOWKHUDS\QRQ-fasted state allowed); and
x
once at the EOS.
p. CRP laboratory tests will be performed at the following time points:
x
on Day -1;
x
once during ambulatory visit on Day 7; 
x
once on Day 10 (upon admission to clinical unit for confinement);
x
RQFHZLWKLQKDIWHUILUVW3&5SDUDVLWHVP/
x
RQFHRQ'D\RIILUVW3&5SDUDVLWHVP/$1'SULRUWRLQLWLDWLQJDQWLPDODULDOWKHUDS\
x
on Days 2 and 3 of antimalarial therapy;
x
once at pre-GLVFKDUJHIURPFOLQLFDOXQLWRQ'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'KDIWHULQLWLDWLQJDQWLPalarial therapy); and
x
once at the EOS.
q. INR, PT and aPTT laboratory tests will be performed once at screening and once on Day -1. On the other time points thereafter, only the INR test will be performed.
r. A nasopharyngeal swab will be taken at an ambulatory visit during screening up to Day -2 (preferably as close as possible to the day of admission, based on test capacity and 
according to national laws) from participants who are still eligible to be enrolled in the study after completing all other screening tests. An rRT-PCR test will be performed on these 
swabs to screen for infection with SARS-CoV-2.
s. A nasopharyngeal swab will be taken prior to confinement to the study site on Day 10, based on test capacity and according to national laws.
t. Blood samples for the assessment of parasitaemia by qPCR will be drawn at the following time points:
x
on Day 1 (pre-PfSPZ-DVI Challenge);
x
once daily during ambulatory visits on Day 7, 8 and 9;
x
twice daily from Day 10 (admission to clinical unit for confinement) onwards until Day of ILUVW3&5SDUDVLWHVP/$1'SULRUWR initiating antimalarial therapy;
x
at multiple time points (2, 6, 8, 12, 16, 24, 36, 48 and 72 h) following initiation of Riamet®;
x
RQFHRQ'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'KDIWHULQLWLDWLQJDQWLPDODULDOWKHUDS\; and
x
once at the EOS.

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01
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33
All qPCR blood samples will be drawn for immediate analysis, except for the ones drawn at the multiple time points during Riamet® therapy.
u. Blood samples for parasite transcriptomics will be drawn at the following time points:
x
once daily during ambulatory visits on Day 7, 8 and 9;
x
once daily from Day 10 (admission to clinical unit for confinement) onwards until Day of discharge from confinement (Day of pDUDVLWHFOHDUDQFH$1'KDIWHULQLWLDWLQJ
antimalarial therapy); and
x
once at the EOS.
v. All participants will be asked non-leading questions to determine the occurrence of any AEs at the EOS visit.
Of note, for participants who commence antimalarial therapy on Day 24 (l), the Day of discharge from confinement will coincide with the EOS. In this case, applicable assessments will 
not be performed in duplicate.

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Cohort 2
Assessments
Screening Visit
Day -28 to Day -2
Challenge, treatment and follow-up
EOS Visit
Day 28v
Study Day/Event
Day -1a
Day 1
Day 2-6
Day 7-9
Day 10: Start 
of 
Confinement 
in Clinical 
Unit
Day of First PCR 
 Parasites/mL Until 
Day of First PCR 
 Parasites/mL
Day of First PCR 
 Parasites/mL Until 
 h After Initiating 
Antimalarial Therapy
Day of Discharge From 
Confinement: Day of 
Parasite Clearance 
(0 Parasites/mL) AND 
K$IWHU,QLWLDWLQJ
Antimalarial Therapy
Time point (h) in 
relation to 
PfSPZ-DVI 
Challenge
0b
2
Estimated On or Before 
Day 22
Ambulatory visit
X
X
X
Ambulatory visit 
(SARS-CoV-2 test)
X
X
Confinement in 
clinical unitc
X
X
X
X
X
(X)
Discharge from 
clinical unit
X
X
(X)
Telephonic 
Monitoring for AEs
X
Eligibility criteria
X
X
Informed consentd
X
Demographics
X
Medical and social 
history
X
Beck Depression 
Inventory
X
Alcohol & drug 
screene
X
X
Height & weightf
X
X
X
Physical 
examinationg
X
X
X
X
X
X
X
Vital signsh
X
X
X
X
X
X
X
X
X
12-lead ECGi
X
X
Viral serologyj
X
Pregnancy testk
X
X
X
DVI of PfSPZ 
Challenge
X
Riamet®l
Xl

REVISED CLINICAL STUDY PROTOCOL - MMV_PfSPZ-DVI Blood Stage_19_01
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Assessments
Screening Visit
Day -28 to Day -2
Challenge, treatment and follow-up
EOS Visit
Day 28v
Study Day/Event
Day -1a
Day 1
Day 2-6
Day 7-9
Day 10: Start 
of 
Confinement 
in Clinical 
Unit
Day of First PCR 
 Parasites/mL Until 
Day of First PCR 
 Parasites/mL
Day of First PCR 
 Parasites/mL Until 
 h After Initiating 
Antimalarial Therapy
Day of Discharge From 
Confinement: Day of 
Parasite Clearance 
(0 Parasites/mL) AND 
K$IWHU,QLWLDWLQJ
Antimalarial Therapy
Time point (h) in 
relation to 
PfSPZ-DVI 
Challenge
0b
2
Estimated On or Before 
Day 22
Malaria Clinical 
Scorem
X
X
X
X
X
X
X
Haematology &
liver biochemistryn
X
X
X
X
X
X
X
X
Biochemistry other 
than livero
X
X
X
X
X
CRP analysisp
X
X
X
X
X
X
X
Coagulation 
parametersq
X
X
X
X
X
X
X
X
rRT-PCR for 
SARS-CoV-2
Xr
Xs
qPCR for parasitest
X
X
X
X
X
X
X
Parasite 
Transcriptomicsu
X
X
X
X
X
X
Previous 
medications
X
Concomitant 
medications
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X
AEs
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X
SAE reporting
X---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------X
ȕ-+&*ȕ-human chorionic gonadotropin, AE: adverse event, aPTT: activated partial thromboplastin time, CMV: cytomegalovirus, CRP: C-reactive protein, DVI: direct venous 
inoculation, EBV: Epstein Barr Virus, ECG: electrocardiogram, EOS: end of study, FSH: follicle stimulating hormone, HbsAg: hepatitis B surface antigen , HCV: hepatitis C virus, 
HIV: human immunodeficiency virus, IgM: immunoglobulin M, INR: international normalized ratio, PfSPZ: Plasmodium falciparum sporozoites, PT: prothrombin time, 
qPCR: quantitative polymerase chain reaction, rRT-PCR: real-time reverse transcription polymerase chain reaction,  SAE: serious adverse event, SARS-CoV-2: severe acute respiratory 
syndrome coronavirus 2
a. Assessments begin as of participant confinement in the clinical unit and must be completed and safety assessment outcomes must be available before PfSPZ-DVI Challenge on
Day 1.
b. On Day 1, assessments begin as of 3 h before challenge (except for urinalysis assessments, morning void allowed). The assessments indicated must be completed and safety 
assessment outcomes must be available before PfSPZ-DVI Challenge inoculation.

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c. Confinement to the clinical unit will occur as follows:
x
Participants will be admitted to the clinical unit in the morning of Day -1 and discharged from confinement 2 h post PfSPZ-DVI Challenge on Day 1.
x
Participants will be admitted to the clinical unit in the morning of Day 10 and discharged on Day of discharge from FRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'KDIWHU
initiating antimalarial therapy), estimated to occur on or before Day 22. Of note, participants who commence antimalarial therapy on Day 24 (see l) will only be discharged from 
confinement at the end of the EOS visit on Day 28.
d. No study-related procedure is to be performed before voluntarily signing of the informed consent form.
e. Alcohol breath test and urine dipstick screening for drug abuse.
f. Height to be measured at screening only; body weight to be measured at screening, at admission on Day -1 and at the EOS visit.
g. Physical examination will be conducted:
x
once at screening and once on Day -1;
x
once on Day 10 (upon admission to clinical unit for confinement); 
x
RQFHZLWKLQKDIWHUILUVW3&5SDUDVLWHVP/
x
once ZLWKLQKDIWHUILUVW3&5SDUDVLWHVP/
x
once at pre-discharge from clinical unit on Day of discharge from confinement (Day of parasite clearance $1'KDIWHULQLWLDWLQJDQWLPDODULDOWKHUDS\); and
x
once at the EOS. 
Full physical examination will be conducted at screening, on Day -ZLWKLQKDIWHUILUVW3&5SDUDVLWHVP/DQGDWWKH(26YLVLWRQ'D\7DUJHWHGV\PSWRP-driven) 
physical examination will be performed on the other time points in case malaria clinical score >6.
h. Vital signs (blood pressure and pulse) will be measured after remaining in a supine position for at least 10 min. All measurements will be performed as follows:
x
once at screening and once on Day -1;
x
on Day 1 (pre and post PfSPZ-DVI Challenge);
x
twice daily from Day 10 (DGPLVVLRQWRFOLQLFDOXQLWIRUFRQILQHPHQWRQZDUGVXQWLO'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'2 h after initiating 
antimalarial therapy); and
x
once at the EOS.
Body temperature (sublingual) will also be assessed, and this whenever the malaria clinical score will be evaluated (see m).
At screening, orthostatic changes to blood pressure and pulse rate will also be assessed: participants will be requested to stand after completion of the supine measurements and 
blood pressure and pulse rate will be recorded after at least 2 min in the standing position.
i. 12-lead ECGs recordings will be performed in triplicate after participants have remained in a supine position for at least 10 min. All recordings will be performed as follows:
x
once at screening; and
x
once on Days 2 and 3 of antimalarial therapy.
j. Serological testing for HIV, hepatitis A IgM antibody, HbsAg, anti-HCV antibody, hepatitis D antibody (only in subjects positive for HbsAg), hepatitis E IgM antibody, CMV IgM 
antibody and EBV IgM antibody, to determine eligibility for the study.
k. 3UHJQDQF\WHVWLQJFRQVLVWVRIVHUXPȕ-HCG assessments at screening and at the EOS visit and a XULQHȕ-HCG assessment on Day-1.
l. Every participant will be prescribed a 3-day oral course of Riamet® (20 mg artemether and 120 mg lumefantrine tablets) (see Section 5.1). The first dose should be taken as soon as 
possible when a PCR threshold of 10000 parasites/mL is achieved or earlier if a participant has a malaria clinical score >6 or at Investigator’s discretion. Of note, all participants 
inoculated with PfSPZ-DVI Challenge will commence antimalarial therapy no later than Day 24, regardless if they reach the pre-defined cohort-specific PCR parasitaemia/malaria 
FOLQLFDOVFRUHWKUHVKROGVLHSDUDVLWHVP/EORRGDQGRUDPDODULDFOLQLFDOVFRUHVHH6HFWLRQ 4.3).
If an intolerance or contraindication to Riamet® develops since inclusion, Malarone® will be administered (see Section 5.1).
m.Malaria clinical score for malaria signs and symptoms and the vital sign parameter body temperature (sublingual) (see h) are assessed:
x
on Day 1 (pre-PfSPZ-DVI Challenge); 
x
once daily during ambulatory visits on Day 7, 8 and 9;
x
twice daily from Day 10 (admission to clinical unit for confinement) onwards until Day of discharge from confinement (Day of parasiWHFOHDUDQFH$1'2 h after initiating 
antimalarial therapy); and
x
once at the EOS.

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n. Haematology and liver biochemistry laboratory tests will be performed after overnight fast for at least 8 h (see Section 4.3):
x
once at screening and once on Day -1;
x
once during ambulatory visit on Day 7; 
x
once on Day 10 (upon admission to clinical unit for confinement);
x
RQFHZLWKLQKDIWHUILUVW3&5SDUDVLWHVP/
x
once on Day of first 3&510000 parasites/mL AND prior to initiating antimalarial therapy (non-fasted state allowed);
x
once at pre-discharge from clinical unit RQ'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'KDIWHULQLWLDWLQJDQWLPDODULDOtherapy); and
x
once at the EOS.
Troponin T will be measured at screening (fasted state), at baseline prior to inoculation on Day -1 (fasted state), and on Days 1, 2 and 3 of antimalarial therapy (non-fasted state).
FSH will be measured at screening in all women.
o. Biochemistry laboratory tests other than liver (including urinalysis; morning void allowed) will be performed after overnight fast for at least 8 h (see Section 4.3):
x
once at screening and once on Day -1;
x
once on Day 10 (upon admission to clinical unit for confinement);
x
RQFHRQ'D\RIILUVW3&5SDUDVLWHVP/$1'SULRUWRLQLWLDWLQJDQWLPDODULDOWKHUDS\QRQ-fasted state allowed); and
x
once at the EOS.
p. CRP laboratory tests will be performed at the following time points:
x
on Day -1;
x
once during ambulatory visit on Day 7; 
x
once on Day 10 (upon admission to clinical unit for confinement);
x
RQFHZLWKLQKDIWHUILUVW3&5SDUDVLWHVP/
x
oncHRQ'D\RIILUVW3&5SDUDVLWHVP/$1'SULRUWRLQLWLDWLQJDQWLPDODULDOWKHUDS\
x
on Days 2 and 3 of antimalarial therapy;
x
once at pre-GLVFKDUJHIURPFOLQLFDOXQLWRQ'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'KDIWHULQLWLDting antimalarial therapy); and
x
once at the EOS.
q. INR, PT and aPTT laboratory tests will be performed once at screening and once on Day -1. On the other time points thereafter, only the INR test will be performed.
r. A nasopharyngeal swab will be taken at an ambulatory visit during screening up to Day -2 (preferably as close as possible to the day of admission, based on test capacity and 
according to national laws) from participants who are still eligible to be enrolled in the study after completing all other screening tests. An rRT-PCR test will be performed on these 
swabs to screen for infection with SARS-CoV-2.
s. A nasopharyngeal swab will be taken prior to confinement to the study site on Day 10, based on test capacity and according to national laws.
t. Blood samples for the assessment of parasitaemia by qPCR will be drawn at the following time points:
x
on Day 1 (pre-PfSPZ-DVI Challenge);
x
once daily during ambulatory visits on Day 7, 8 and 9;
x
twice daily from Day 10 (admission to clinical unit for confinement) onwards XQWLO'D\RIILUVW3&5SDUDVLWHVP/$1'SULRUWRLQLWLDWLQJDQWLPDODULDOWKHUDS\
x
at multiple time points (2, 6, 8, 12, 16, 24, 36, 48 and 72 h) following initiation of Riamet®;
x
RQFHRQ'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'KDIWHULQLWLDWLQJDQWLPDODULDOWKHUDS\DQG
x
once at the EOS.
All qPCR blood samples will be drawn for immediate analysis, except for the ones drawn at the multiple time points during Riamet® therapy.
u. Blood samples for parasite transcriptomics will be drawn at the following time points:
x
once daily during ambulatory visits on Day 7, 8 and 9;
x
once daily from Day 10 (adPLVVLRQWRFOLQLFDOXQLWIRUFRQILQHPHQWRQZDUGVXQWLO'D\RIGLVFKDUJHIURPFRQILQHPHQW'D\RISDUDVLWHFOHDUDQFH$1'h after initiating 
antimalarial therapy); and
x
once at the EOS.

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v. All participants will be asked non-leading questions to determine the occurrence of any AEs at the EOS visit.
Of note, for participants who commence antimalarial therapy on Day 24 (l), the Day of discharge from confinement will coincide with the EOS. In this case, applicable assessments will 
not be performed in duplicate.

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LIST OF ABBREVIATIONS AND DEFINITIONS OF TERMS
List of Abbreviations
ABV
Alcohol by volume
AE
Adverse event
AESI
Adverse event of special interest
ALP
Alkaline phosphatase
ALT
Alanine aminotransferase
aPTT
Activated partial thromboplastin time
AST
Aspartate aminotransferase
ȕ-HCG
ȕ-human chorionic gonadotropin
BMI
Body mass index
BSC
Biosafety cabinet
CHMI
Controlled Human Malaria Infection
CI
Confidence interval
CMV
Cytomegalovirus
CRP
C-reactive protein
CTCAE
Common Terminology Criteria for Adverse Events
DBP
Diastolic blood pressure
DVI
Direct venous inoculation
EBV
Epstein Barr Virus
ECG
Electrocardiogram
EOS
End of study
FSH
Follicle stimulating hormone
GCP
Good Clinical Practice
HBsAg
Hepatitis B surface antigen
HBV
Hepatitis B virus
HCV
Hepatitis C virus
HIV
Human immunodeficiency virus
HSA
Human serum albumin
IBSM
Induced blood-stage malaria
ICF
Informed Consent Form
ICH
International Council for Harmonization of Technical Requirements 
for Pharmaceuticals for Human Use
IEC
Independent Ethics Committee
IgM
Immunoglobulin M
IMP
Investigational medicinal product
INR
International normalized ratio
IRB
Institutional Review Board
ITM
Centre of excellence for tropical medicine
Log10PRR48
Log10 parasite reduction ratio per 48 h 
Log10PRR48,max
Maximum log10 parasite reduction ratio per 48 h
LPLV
Last Patient Last Visit
MedDRA
Medical Dictionary for Regulatory Activities
NCE
New chemical entities
NSAID
Non-steroidal anti-inflammatory drug
PBS
Phosphate buffered saline

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PC½
Parasite clearance half-life
PCR
Polymerase chain reaction
PD
Pharmacodynamics
Pf
P. falciparum 
P. falciparum
Plasmodium falciparum
PfSPZ
P. falciparum sporozoites
PfSPZ-DVI
Challenge
Inoculation of P. falciparum sporozoites through direct venous 
inoculation
PI
Principal Investigator
PMR
Parasite multiplication rate
PMR48
Parasite multiplication rate standardised to 48 h
PMRLC
Parasite multiplication rate per life cycle (if not 48 h)
PRR
Parasite reduction ratio
PT
Prothrombin time
qPCR
Quantitative polymerase chain reaction
QTc
Corrected QT interval 
QTcB
QT interval corrected according to Bazett’s formula
QTcF
QT interval corrected according to Fridericia’s formula
RBC
Red blood cell
rRT-PCR
Real-time reverse transcription polymerase chain reaction
SAE
Serious adverse event
SARS-CoV-2
Severe acute respiratory syndrome coronavirus 2
SBP
Systolic blood pressure
SD
Standard deviation
SmPC
Summary of Product Characteristics
SPZ
Sporozoites
SRT
Safety Review Team
TBS
Thick Blood Smear
ULN
Upper Limit of Normal
varATS
The acidic terminal segment in Plasmodium falciparum var genes
VIS
Volunteer Infection Study
WBC
White blood cell
WHO
World Health Organization

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Definitions of Terms
BMI
Weight in kilogram divided by the square of height in meters
QTcB
QT interval corrected according to Bazett’s formula: 
QTcB (ms)=QT (ms)/RR1/2 where RR=(60/heart rate)*1000
QTcF
QT interval corrected according to Fridericia’s formula: 
QTcF (ms)=QT (ms)/RR1/3 where RR=(60/heart rate)*1000
Laboratory-defined 
PCR positivity
qPCR outcome 50-249 parasites per mL blood
Protocol-defined PCR 
positivity
qPCR outcome 250 parasites per mL blood
PCR negative
Applies to all qPCR assessments undertaken prior to initiating 
antimalarial therapy, with a value of 1-49 parasites per mL 
blood
Parasite clearance
Defined as a qPCR value of 0 parasites per mL blood after 
initiating antimalarial therapy
Time to first PCR
positivity
Time elapsed between PfSPZ-DVI Challenge and first qPCR 
outcome equal to or greater than 250 parasites per mL blood, 
with a maximum of 28 days in the absence of parasitaemia
Time to parasitaemia 
of 5000 parasites per 
mL blood
Time elapsed between PfSPZ-DVI Challenge and first qPCR 
equal to or greater than 5000 parasites per mL blood, with a 
maximum of 28 days in the absence of parasitaemia
Time to parasitaemia 
of 10000 parasites 
per mL blood
Time elapsed between PfSPZ-DVI Challenge and first qPCR 
equal to or greater than 10000 parasites per mL blood, with a 
maximum of 28 days in the absence of parasitaemia
Parasite growth rate
Expressed as the PMR48 or as the PMRLC
Time to parasite 
clearance
Time between drug administration and first PCR below the 
limit of quantification
Log10PRR48
Ratio of the parasite density at a specific time point to the 
parasite density 48 h later and expressed in log10
Log10PRR48,max
Maximum log10PRR48
PC½
Time taken for the parasite density to be reduced by 50% 
after the first dose administration of Riamet®

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STUDY ADMINISTRATIVE STRUCTURE AND 
INVESTIGATORS
SPONSOR
Medicines for Malaria Venture (MMV)
ICC – Building G
20, route de Pré-Bois
P O Box 1826
1215 Geneva 15
Switzerland
Medical and Project Director:
Farouk Chughlay
Clinical Trial Manager:
Julia Flynn
Clinical Scientist:
Amina Haouala
CLINICAL CENTRE
SGS Life Sciences, Clinical Pharmacology 
Unit
Lange Beeldekensstraat 267
2060 Antwerpen
Belgium
Tel: +32 (0)3 217 25 60
Fax: +32 (0)3 217 25 81
Principal Investigator:
Pieter-Jan Berghmans
CLINICAL MONITORING
IQVIA RDS & Integrated Services Belgium 
NV/SA
Corporate Village, Davos Building
Leonardo Da Vincilaan 7 
1930 Zaventem
Belgium
MEDICAL MONITORING
ICON Clinical Research
Heinrich-Hertz Str. 26 
63225 Langen
Germany
CLINICAL LABORATORY
ZNA Klinisch Laboratorium
Campus Middelheim
Lindendreef 1
2020 Antwerpen
Belgium
BIOANALYTICAL LABORATORY
Institute of Tropical Medicine Antwerp
Nationalestraat 155
2000 Antwerpen
Belgium

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PHARMACOVIGILANCE
PrimeVigilance Ltd. (Head office)
1 Occam Court
Surrey Research Park
Guildford, Surrey GU2 7HJ
United Kingdom

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1. INTRODUCTION
1.1
MALARIA BACKGROUND INFORMATION
Malaria is an infectious disease that threatens half of the world’s population and is 
caused mainly by two protozoan parasites: Plasmodium falciparum (P. falciparum) and 
Plasmodium vivax. The World Health Organization (WHO) reported 219 million cases of 
the disease in 2017. In the same year, malaria caused 435000 deaths; 93% of the 
mortality was in Africa, with children <5 years of age attributing 61.1% of deaths [29].
Around the world, effective disease control programs relying on artemisinin-containing 
combination therapies (ACT) such as Coartem®/Riamet® have contributed to a global 
reduction in the mortality rate of P. falciparum malaria. However, recent reports suggest 
that decades of continuous use of artemisinins as monotherapies may have fostered drug 
resistance against artemisinin-derivatives, the last widely effective antimalarial drugs 
[28].
Malaria is a serious and life-threatening disease and due to increasing resistance of the 
current antimalarial therapies, new antimalarial therapies are required for both treatment 
and prevention of the disease to address a growing unmet medical need.
1.2
MALARIA CHMI/VIS MODELS
Volunteer Infection Studies (VIS) in malaria (previously referred to as Controlled 
Human Malaria Infection (CHMI) studies) are a valuable tool in antimalarial drug 
development, providing an opportunity to rapidly assess drug efficacy and obtain 
pharmacokinetic and pharmacodynamic (PD) data in healthy, non-immune volunteers 
and earlier entry into patients. VIS entail deliberate infection with malaria parasites, 
either by inoculation of sporozoites through mosquito bite or direct injection of 
sporozoites (i.e., inoculation of P. falciparum sporozoites through direct venous 
inoculation, PfSPZ-DVI Challenge) or by direct venous injection of parasitised 
erythrocytes (i.e., Induced Blood-Stage Malaria, IBSM). VIS has been applied in malaria 
vaccine and drug development, where it is used to evaluate products in well-controlled 
early-phase proof-of-concept clinical studies, thus facilitating progression of only the 
most promising candidates for further evaluation in areas where malaria is endemic. 
1.3
PLASMODIUM FALCIPARUM SPOROZOITES CHALLENGE MODEL 
(PFSPZ CHALLENGE)
Recent advances to support vaccine and drug development have included 
cryopreservation of sporozoites (PfSPZ-DVI Challenge model). PfSPZ-DVI Challenge 
consists of aseptic, purified, live, fully infectious, cryopreserved (to maintain potency for 
an extended period) Plasmodium falciparum (Pf) sporozoites (SPZ) (PfSPZ) obtained 
from the salivary glands of Anopheles stephensi mosquitoes reared under aseptic 
conditions and infected with PfSPZ of either the NF54 strain or 7G8 clone. This advance 
will help to accelerate malaria vaccine and drug development by making the reagents for 
VIS more widely accessible and enabling a more rigorous evaluation with multiple 
parasite strains and species.
The PfSPZ-DVI Challenge agent was developed to be used to infect Phase I volunteers in 
VIS to assess the protective efficacy or efficacy of antimalarial drugs and vaccines, and 

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the effects of naturally acquired immunity and innate resistance. By facilitating the 
development of liver stage infection, PfSPZ-DVI Challenge can be utilized in the 
assessment of causal prophylactic activity of drugs, in contrast with IBSM model which 
results in blood-stage infection only, and consequently, for only assessing drug activity 
against the blood stage of malaria. As is the case with IBSM, the PfSPZ-DVI Challenge 
model, which has been extensively used for testing vaccine and drug prevention 
strategies, has the potential to also be utilised in the assessment of blood-stage activity of 
new antimalarial drugs, and thus increasing the existing MMV and MMV partners’ 
capability for conducting blood-stage malaria VIS at various study centres during early 
drug development of new chemical entities (NCEs).
Conducting VIS in antimalarial vaccine and drug studies with PfSPZ-DVI Challenge as 
compared to P. falciparum sporozoite-infected mosquitoes or parasitised red blood cells
(RBCs) (IBSM model) provides the following potential advantages: 
1. Capacity to conduct clinical studies at multiple sites that do not have access to 
PfSPZ-infected mosquitoes or a Master Cell Bank with parasitised erythrocytes;
2. Increased capacity to conduct VIS critical to the development of NCEs/drugs for 
acute treatment of blood-stage malaria;
3. Capacity to determine and control the specific numbers of PfSPZ with which 
volunteers are inoculated;
4. Capacity to eliminate the potential impact of batch to batch variation on 
infectivity of sporozoites when performing parallel clinical studies at multiple 
sites or sequential clinical studies at the same site
5. Capacity to conduct VIS in volunteers without the study schedule being 
contingent upon a time-consuming and financially expensive process for 
producing infected mosquitoes that must be used within a one to two-week 
window. Benefits include the ability to perform repeated low-dose VIS for time-
course studies with vaccines and drugs, to stagger the VIS for different groups, to 
“catch-up” volunteers that miss the scheduled VIS day, and to use adaptive study
designs where the date and nature of VIS can be made dependent on outcome 
variables measured during the study;
6. Potentially significantly reduced costs.
PfSPZ-DVI Challenge has been safely used in multiple clinical studies in the United 
States, Europe and Africa via the intradermal (ID), intravenous (IV) and intramuscular 
(IM) routes [1,2,7,8,10,11,18,19,23,24,25,26]. As of 17 April 2018, 913 volunteers have 
received 1439 doses of PfSPZ Challenge (NF54) and 34 volunteers have received 
34 doses of PfSPZ Challenge (7G8). Extensive safety and efficacy data are available [9]. 
In addition, PfSPZ-DVI Challenge has been safely administered to volunteers in 
combination with antimalarial drugs and to assess the effects of naturally acquired 
immunity and innate resistance (e.g., sickle cell trait). Currently, the validated 
inoculation method for drug and vaccine evaluation consists of direct venous inoculation 
(DVI) with an inoculum size of 3200 sporozoites which has been shown to induce 
subclinical malaria in 100% of inoculated healthy volunteers [18].
For more detailed information on the PfSPZ-DVI Challenge (NF54 strain) used in this 
study, refer to the current IB [9]. 

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1.4
ANTIMALARIAL THERAPY
The registered antimalarial therapy Riamet® (tablet containing 20 mg artemether and 
120 mg lumefantrine) has been approved for the treatment of acute uncomplicated 
P. falciparum PDODULDLQIHFWLRQLQDGXOWVFKLOGUHQDQGQHZERUQVZHLJKLQJNJ)RU
adult patients, the treatment regimen consists of 6 doses of 4 tablets administered over a 
period of 60 h (1 intake at diagnosis and the 5 following doses at 8, 24, 36, 48 and 60 h
after the initial dose). This Riamet® therapeutic regimen was tested in the IBSM model in 
Australia [15] and will also be used in this study.
In adults, the most frequently reported adverse reactions are decreased appetite, sleep 
disorders, headache, dizziness, palpitations, vomiting, abdominal pain, nausea, arthralgia 
DQGP\DOJLDUHSRUWHGLQSDWLHQWV>YHU\FRPPRQ@DQGLQVRPQLDSDUHVWKHVLD
clonus electrocardiogram (ECG) QT prolonged, diarrhoea, rash and pruritus (reported in 
WR patients [common]). 
For more information on Riamet®, refer to the Summary of Product Characteristics 
(SmPC) [22].
1.5
OVERALL RATIONALE FOR THE STUDY
The intent of this study is to evaluate the feasibility of PfSPZ-DVI Challenge in the 
assessment of NCEs and drugs for the acute treatment of blood-stage malaria. Unlike 
malaria chemoprotection studies where study participants are treated early with rescue 
medication, the intention of utilising the PfSPZ-DVI Challenge for the assessment of 
NCEs/drugs for the acute treatment of blood-stage malaria, is to safely and reliably 
generate blood-stage malaria with appropriate/acceptable parasitaemia densities in 
healthy participants to generate key data supporting further dose selection for patient 
studies in the field. To our knowledge the PfSPZ-DVI Challenge model has not been 
used for assessing antimalarial blood-stage activity in an early-drug development 
context, and a study in healthy participants is needed to determine its safety/tolerability 
and feasibility in assessing antimalarial blood-stage activity.
Existing evidence in support of this methodology in achieving blood-stage parasitaemia
in healthy participants, whilst demonstrating acceptable safety and tolerability is based 
on published literature [7,18,10,2].
1.6
RISK BENEFIT ANALYSIS
1.6.1
Potential Risks
Participants receiving the PfSPZ-DVI Challenge will be exposed to potential risks, 
including the development of malaria symptoms as well as adverse effects of the 
registered antimalarial therapy (Riamet® or Malarone®; see SmPCs [22] and [12]) to be 
prescribed as curative therapy for parasite clearance prior to the end of the study.
The VIS PfSPZ-DVI Challenge model with the DVI technique is a well-validated 
method for obtaining informative data on the clinical activity of novel antimalarial 
compounds. It has replaced the traditional method of CHMI by the bite of mosquitos to 
measure vaccine and drug efficacy. In a series of clinical studies, inoculation of 
PfSPZ-DVI Challenge was found to be safe and well tolerated with the optimal dose 
established as 3200 PfSPZ administered by DVI [18].

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Overall, the PCR (Polymerase chain reaction) monitoring used in these studies ensure 
early treatment preventing high levels or prolonged duration of parasitaemia that would 
put the participant at undue risk. Severe/complicated malaria has never been reported in a 
VIS. Mild malaria symptoms are expected in some study participants and include 
headache, myalgia, fever, chills, sweats, nausea, vomiting and diarrhoea. Rapid diagnosis 
by quantitative PCR (qPCR) and subsequent treatment by Riamet® in the study is 
expected to quickly attenuate the illness so that the infection does not place the 
participant at undue risk. 
Risks to participants in this Phase Ib study will be minimised in the following ways:
-
Sentinel strategy for both Cohort 1 and 2 (please refer to Section 3.1);
-
Progression to consecutive study Cohort 2 only after evaluation of data from 
previous Cohort 1 and approval from Safety Review Team (SRT) (please refer 
to Section 3.4);
-
Adherence to the inclusion/exclusion criteria: only participants considered 
suitable according to these criteria and who are not at any perceived risk will be 
included;
-
The dose of PfSPZ-DVI Challenge and route of administration has previously 
been shown to be safe and well-tolerated in healthy human participants
[2,7,10,18];
-
Routine monitoring to ensure the safety and well-being of the study participants
will include the following:
o
Admission to the clinical unit in the morning of Day -1, i.e. before 
PfSPZ-DVI Challenge, until 2 h post PfSPZ-DVI Challenge and again 
from Day 10 in the morning until parasite clearance (defined as a qPCR 
value of 0 parasites per mL blood after initiating antimalarial therapy) and 
at least 72 h after initiating antimalarial therapy or until Day 28 when 
antimalarial therapy initiated on Day 24;
o
Vital signs (blood pressure, heart rate);
o
12-lead ECGs;
o
Safety laboratory assessments of biochemistry and haematological blood 
parameters performed at scheduled time points and repeated, if necessary
(including repeat of any alanine aminotransferase [ALT] or aspartate
aminotransferase [AST] >2x ULN), to ensure appropriate follow-up of 
any clinically relevant abnormality;
o
Daily parasitaemia assessments by qPCR during daily visits to the clinical 
unit from Day 7 until Day 9 and twice daily from Day 10 post inoculation 
until parasite clearance and at the EOS visit at Day 28;
o
Objective/directed evaluation of Malaria signs and symptoms through a 
clinical score.
-
All participants will be prescribed curative therapy for malaria for radical 
clearance during the study;
-
The total volume of blood drawn per the Time and Events Schedule from each 
participant will not exceed the maximum allowable volume of approximately 
460 mL;
-
In addition to the Principal Investigator (PI) or his representative, the site has 
access to an expert medical malariologist to help in making clinical decisions as 

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necessary, including in the rare event that hospitalisation is required, which will 
be done at the Antwerp University Hospital (UZA). Serious adverse event 
(SAEs) will be treated according to their nature at a specialist inpatient facility. 
1.6.2
Potential Benefits
Benefits of the study are society-based and related to contributing to the future 
development of new antimalarial therapies. Taking into consideration the proposed risk-
management plan as described, the risk to participants participating in this study is 
considered to be minimal and acceptable.
1.6.3
Conclusion
No benefit is expected for individual participants participating in this study. Benefits of 
the study are society-based and related to possible future antimalarial therapies that 
would be developed using this VIS model. Taking into consideration the proposed 
risk-management plan as described, the risk to participants participating in this study is 
considered to be minimal and acceptable.

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2. OBJECTIVES AND ENDPOINTS
Table 1:
Objectives and Endpoints
OBJECTIVES
ENDPOINTS
For definitions of terms, see List of 
Abbreviations and Definitions of Terms
Primary
x
To evaluate the safety and 
tolerability associated with 
blood-stage malaria infection in 
naïve healthy participants after 
PfSPZ-DVI Challenge.
x
The incidence and severity of observed 
(i.e., malaria signs and symptoms graded 
by the malaria clinical score; see 
Attachment 1) or self-reported adverse 
events (AEs) considered PfSPZ-DVI 
Challenge inoculum-related until end of 
study (EOS) visit.
x
The change in malaria clinical score from 
PfSPZ-DVI Challenge until parasite 
clearance.
x
Changes from baseline in haematology, 
clinical chemistry and urinalysis 
parameters, vital signs and ECG 
parameters.

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OBJECTIVES
ENDPOINTS
For definitions of terms, see List of 
Abbreviations and Definitions of Terms
Primary
x
To characterise key stages in the 
blood-stage parasite growth profile 
in malaria infection in naïve 
healthy participants after 
PfSPZ-DVI Challenge.
x
The parasite growth profiles of 
P. falciparum NF54 after infection via 
injection of sporozoites by DVI will be 
characterised by: 
o time to first PCR positivity (for the 
purpose of this study, ‘PCR positivity’ 
is used for the ‘protocol-defined PCR 
positivity’, unless mentioned 
otherwise);
o parasitaemia at first PCR positivity;
o time to parasitaemia of 
5000 parasites per mL blood 
(Cohorts 1 and 2);
o parasitaemia at the time parasitaemia 
SDUDVLWHVSHUP/EORRG
(Cohorts 1 and 2);
o time to parasitaemia of 
10000 parasites per mL blood 
(Cohort 2);
o parasitaemia at the time parasitaemia 
SDUDVLWHVSHUP/EORRG
(Cohort 2);
o time to first dose of treatment with 
artemether-lumefantrine (Riamet®)
(Cohorts 1 and 2); and
o parasitaemia at first dose of treatment 
with Riamet® (Cohorts 1 and 2).
x
The number and proportion of participants 
with presence of positive PCR and 
SDUDVLWDHPLDRIRUSDUDVLWHV
per mL blood between inoculation with 
PfSPZ-DVI Challenge and Day 28 (per 
cohort).

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OBJECTIVES
ENDPOINTS
For definitions of terms, see List of 
Abbreviations and Definitions of Terms
Secondary
x
To determine the safety and 
tolerability of Riamet® in 
blood-stage malaria infection in 
naïve healthy participants after 
PfSPZ-DVI Challenge.
x
The incidence, severity and relationship to 
Riamet® of observed or self-reported, 
treatment-emergent adverse events 
(TEAEs) until EOS visit.
x
To characterise the blood-stage 
parasite profile in malaria infection 
in naïve healthy participants after 
PfSPZ-DVI Challenge.
x
The blood-stage parasite profile of 
P. falciparum NF54 after infection via 
injection of sporozoites by DVI will be 
characterised by: 
o parasite growth rate expressed as the 
parasite multiplication rate (PMR) 
standardised to 48 h (PMR48)
(Cohort 1, Cohort 2 and overall);
o parasite growth rate expressed as the 
PMR per life cycle (if not 48 h) 
(PMRLC) (Cohort 1, Cohort 2 and 
overall); and
o predicted time to reach parasitaemia 
threshold of first positive PCR, 
 parasites per mL blood, 
 parasites per mL blood for 
Cohorts 1 and 2.
x
To characterise the blood-stage 
parasite clearance profile of 
Riamet® in malaria infection in 
naïve healthy participants after 
PfSPZ-DVI Challenge.
x
The effect of Riamet® on clearance of 
P. falciparum blood-stage parasites will 
be evaluated using:
o time to parasite clearance (Cohort 1, 
Cohort 2 and overall);
o log10 parasite reduction ratio per 48 h 
(log10PRR48) (Cohort 1, Cohort 2 and 
overall);
o maximum log10PRR48 (log10PRR48,max)
(Cohort 1, Cohort 2 and overall); and
o parasite clearance half-life (PC½)
(Cohort 1, Cohort 2 and overall).

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OBJECTIVES
ENDPOINTS
For definitions of terms, see List of 
Abbreviations and Definitions of Terms
Exploratory
x
To characterise the individual 
variability of blood-stage parasite 
profile.
x
The individual variability of blood-stage 
parasite profile will be characterised by:
o the inter-individual variability of the 
parasite growth rate; and
o predicted time to reach parasitaemia 
threshold of first positive PCR, 
 parasites per mL blood, 
 parasites per mL blood 
including their 95% confidence 
interval (CI).
HYPOTHESIS: This study is exploratory in nature and therefore no hypothesis will be 
tested.

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3. STUDY DESIGN
3.1
OVERVIEW OF STUDY DESIGN
This is a single-centre, open-label, Phase Ib study designed to assess if intravenous bolus 
injection of approximately 3200 P. falciparum (NF54 strain) sporozoites can be safely 
administered to achieve blood-stage parasitaemia with a kinetics/PCR profile that will 
allow for the future characterisation of antimalarial blood-stage activity of NCEs in a 
relatively small number of participants during early drug development.
Up to 16 healthy, malaria-naïve males and females, aged 18-55 years, will be enrolled in 
a maximum of 2 cohorts (up to 8 participants per cohort; a participant may be enrolled in 
one cohort only). Enrolment into the cohorts will proceed sequentially, with two target 
levels of parasitaemia previously achieved in healthy participants enrolled in malaria VIS 
at other study sites (see Section 3.2), i.e., 5000 parasites/mL blood in Cohort 1 and 
10000 parasites/mL blood in Cohort 2.
Each participant will be admitted to the clinical unit in the morning of Day -1 and
inoculated with approximately 3200 P. falciparum sporozoites (NF54 strain) by DVI on 
Day 1. Participants will be discharged 2 h post inoculation on Day 1 and will be 
monitored daily via phone call from Day 2 until Day 6 to solicit any AEs. Participants
will come to the clinical unit daily from Day 7 until Day 9 and together with the malaria 
clinical score (see Attachment 1), the presence of parasites will be assessed once daily by 
a specific qPCR targeting the varATS (the acidic terminal segment in Plasmodium 
falciparum var genes) multigenic family; this to accurately describe parasite growth even 
in case PCR positivity, i.e., a qPCR RXWFRPHSDUDVLWHVSHUP/EORRG is confirmed 
this early (very low probability and with low densities). Participants will be confined to
the clinical unit from Day 10 in the morning. qPCR will be performed and malaria 
clinical score assessed twice daily and participants will be administered registered 
antimalarial therapy, i.e., Riamet® (see Section 5.1), when the following criteria are met:
1. &RKRUWSDUDVLWHVmL blood or earlier if a participant has a malaria 
clinical score >6 or at Investigator’s discretion.
2. &RKRUWSDUDVLWHVP/blood or earlier if a participant has a malaria 
clinical score >6 or at Investigator’s discretion.
The registered 3-day antimalarial therapy regimen will be further administered and 
monitored. qPCR assessments of parasitaemia will be carried out at multiple time points 
(2, 6, 8, 12, 16, 24, 36, 48 and 72 h) following initiation of Riamet® and malaria clinical 
score will be assessed twice daily during confinement in the clinical unit. Safety and 
tolerability will be monitored during the whole study duration, specific assessments will 
be done at periodic pre-specified time points from Day 10 and for at least 72 h after
initiating antimalarial therapy, i.e., during confinement in the clinical unit (see below).
Of note, all participants must consent to receiving antimalarial therapy, i.e., the registered 
3-day Riamet® regimen approved for treatment of uncomplicated malaria. Even in the 
case of withdrawal from the study, all participants administered the PfSPZ-DVI 
Challenge are to receive antimalarial therapy as soon as possible, and to have all 
appropriate visits and assessments as required. If an intolerance or contraindication to 
Riamet® develops, Malarone® will be administered (see Section 5.1).

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Upon parasite clearance (defined as a qPCR value of 0 parasites per mL blood after 
initiating antimalarial therapy) and at least 72 h after initiating antimalarial therapy
(estimated to occur on or before Day 19 and on or before Day 22 for Cohort 1 and 
Cohort 2, respectively), and if clinically well, participants will be discharged from the 
clinical unit and will be followed up for safety assessments, clinical evaluation and 
malaria qPCR in the clinical unit at the EOS visit on Day 28.
All participants who received antimalarial therapy will be asked non-leading questions to 
determine the occurrence of any AEs throughout the study and at the EOS visit.
All participants inoculated with PfSPZ-DVI Challenge will commence antimalarial 
therapy no later than Day 24 for both cohorts, regardless if they reach pre-defined 
cohort-specific PCR parasitaemia/malaria clinical score thresholds (i.e., 
5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for Cohort 2 and/or 
DPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQGParticipants who start antimalarial 
therapy on Day 24 will only be discharged from confinement at the end of the EOS visit 
on Day 28.
Antimalarial therapy may be initiated whenever deemed necessary by the Investigators, 
e.g., if there is a concern regarding the safety of a study participant (see Section 5.1).
Therapy may be amended according to the treating physician if the participant does not 
respond to treatment or the condition worsens.
A sentinel strategy will be employed in Cohorts 1 and 2. Each cohort will consist of 
2 subgroups of participants, to be enrolled sequentially: Subgroup 1 will be composed of 
2 participants; Subgroup 2 will be composed of 6 participants. Participants in Subgroup 2 
will not be treated until the last participant in Subgroup 1 has completed antimalarial 
therapy and only upon decision of the PI, medical director and medical monitor after 
review of available safety and tolerability data (safety assessments, AEs and malaria 
clinical scores) and parasitaemia data, including clearance of parasitaemia, of 
Subgroup 1.
Review of available Cohort 1 safety/tolerability and parasitaemia data up to the last dose 
of Riamet® for all inoculated participants in Cohort 1 will be performed by an SRT 
before enrolment of Cohort 2 (see Section 3.4). 
A schematic overview of the study design is shown in Figure 1 for Cohort 1 and in 
Figure 2 for Cohort 2. The assessments to be performed are summarised per visit in the 
Time and Events Schedule for Cohort 1 and Cohort 2.

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Figure 1:
Schematic Overview of the Study for Cohort 1
Figure 2:
Schematic Overview of the Study for Cohort 2
3.2
DISCUSSION OF STUDY DESIGN AND DOSE SELECTION
Dose Selection
Currently, the validated inoculation method for drug and vaccine evaluation consists of 
direct venous inoculation (DVI) with an inoculum size of 3200 sporozoites which has 
been shown to induce subclinical malaria in 100% of inoculated healthy volunteers [18].
Based on published data in Phase I participants inoculated with malaria, the registered 
regimen of artemether-lumefantrine (Riamet®) will clear asexual parasitaemia in less 
than 48 h [15].

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Target PCR-Based Parasitaemia Thresholds Informing the Initiation of Antimalarial 
Therapy
The proposed PCR-based parasitaemia thresholds to initiate antimalarial therapy have 
been selected to maximise participant safety whilst ensuring study objectives are met 
through efficient conduct of the study as follows:
1. &RKRUWSDUDVLWHVP/EORRGRUHDUOLHULIDparticipant has a malaria 
clinical score >6 or at Investigator’s discretion.
2. &RKRUWSDUDVLWHVP/EORRGRUHDUOLHULIDparticipant has a malaria 
clinical score >6 or at Investigator’s discretion.
The aim of the proposed clinical study is the evaluation of the feasibility of PfSPZ-DVI 
Challenge to safely generate adequate levels of asexual blood-stage malaria parasitaemia 
for the evaluation of NCEs/drugs for acute treatment of blood-stage malaria. There is 
available evidence from published Phase I validation studies supporting the feasibility of 
this model for this purpose:
1. A Phase I PfSPZ-DVI Challenge study [7,18] was conducted in healthy 
volunteers, including inoculation of 3200 PfSPZ-DVI Challenge in 6 participants
resulting in:
-
Infectivity rate of 100%;
-
Geometric mean time to Thick Blood Smear (TBS)-positive asexual 
parasitaemia of 11.4 days (11.0, 10.4, 12.3, 10.9, 11.9, 12.2 days);
-
Geometric mean time to qPCR-positive asexual parasitaemia of 8.3 days (6.9, 
9.1, 8.0, 8.0, 9.0, 8.9 days);
-
Geometric mean parasite density at time of TBS-positivity (parasites/mL 
blood) of 6600 (5000, 1300, 10000, 8000, 12000, 14000 parasites/mL blood); 
and
-
PfSPZ-DVI Challenge was well tolerated with AEs occurring at the time of 
parasitaemia detection. AEs were mostly grade 1 (51/85) or grade 2 (28/85) in 
severity, with only a few grade 3 AEs reported (6/85). No SAEs occurred.
2. A Phase I study was conducted which assessed standardised VIS by DVI of 
3200 PfSPZ in healthy, adult, lifelong malaria–exposed volunteers [10]. The 
study included 25 participants in total, of which 5 were healthy and 
non-immune/malaria-naïve. Findings in these 5 participants are as follows:
-
Infectivity rate of 100% confirmed by TBS and qPCR;
-
Geometric mean time to TBS-positive parasitemia of 12.6 days 
(range:12-14 days);
-
Geometric mean time to qPCR-positive parasitemia of 7.9 days (range: 
7-9 days);
-
Geometric mean parasite density at first parasitemia of 5000 parasites/mL
blood (range: 2000-9000 mL); and 
-
PfSPZ-DVI Challenge was well tolerated and no serious AE occurred during 
the study. In the 5/5 healthy, malaria-naïve participants, majority of AEs were 
grade 1 (27/50) in severity. A total of 13/50 AEs were grade 2 and a further 
10/50 AEs were grade 3 in severity.

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3. In addition, the chemoprotection MMV_P218_17_01 study conducted in Belgium 
(EudraCT number: 2018-003004-39) [2] data indicate:
-
An infectivity rate of 100%, with asexual parasitaemia detected in 6 out of the 
6 inoculated participants administered placebo. Although the study protocol 
required administration of rescue treatment at the threshold of 
250 parasites/mL blood, 2/6 participants achieved parasite densities of 
1583 and 4432 parasites/mL blood at the time of rescue treatment 
administration; 
-
In Cohort 1 (2 x 1000 mg doses of P218 administered 48 h apart, no 
sporozoite inoculation): No SAEs or severe AEs were reported. A total of 
6 AEs occurred in 3/8 participants, all 6 were grade 1 AEs (mild). Only 
1/6 AEs (diarrhoea, grade 1) was assessed as being related/suspected to be 
due to P218 with the remaining 5/6 deemed not related to P218. In addition, 
no clinically significant findings were noted on vital signs, ECG (no clinically 
concerning prolongation in QT interval corrected according to Fridericia’s 
formula [QTcF] [5]) and laboratory investigations. These findings supported 
the Safety Review Committees’ decision after Cohort 1, to proceed with 
Cohort 2;
-
In Cohort 2 (sporozoite inoculation followed by 2 x 1000 mg doses of P218 
administered 48 h apart): No SAEs or severe AEs were reported. A total of 
26 AEs occurred in 11/12 participants, with 21/26 grade 1 AEs (mild) and 
5/26 grade 2 AEs (moderate). A total of 2/26 AEs in 1 participant (2 headache 
events, grade 1) was assessed as being related to P218 with the remaining 
24/26 AEs deemed not related to P218. A total of 4/26 AEs in 2 participants
(fatigue, malaise, flu-like symptoms and transaminase elevation) was assessed 
as being related to PfSPZ-DVI Challenge. In addition, no clinically significant 
findings were noted on vital signs, ECG (no clinically concerning QTc 
prolongation) and laboratory investigations. These findings supported the 
Safety Review Committees’ decision after Cohort 2, to proceed with 
Cohort 3;
-
In Cohort 3 (sporozoite inoculation followed by 2 x 100 mg doses of P218 
administered 48 h apart): No SAEs or severe AEs were reported. A total of 
7 AEs occurred in 4/12 participants, all were grade 1 AEs (mild). All AEs 
were considered not related to P218. A total of 2/7 AEs in two participants
(both flu-like symptoms) was assessed as being related to PfSPZ-DVI 
Challenge. In addition, no clinically significant findings were noted on vital 
signs, ECG (no clinically concerning QTc prolongation) and laboratory 
investigations; and
-
Of note was the occurrence of a transient, asymptomatic transaminase 
elevation (ALT) >3x the upper limit of normal (ULN) (with LQFUHDVHV[
ULN in AST and no effect on bilirubin) in the participant who achieved a 
peak parasitaemia of 4432 parasites/mL. ALT/AST values were back to 
normal range before study end without specific intervention.
Further indirect evidence in support of the safety rationale for the proposed 
PCR-thresholds is derived from completed MMV-sponsored IBSM studies conducted in 
Australia with the NCEs DSM265 [3] and MMV390048 [14]. The level of parasitaemia 

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to be observed in our proposed pilot study for both Cohorts 1 (5000 parasites/mL) and 2 
(10000 parasites/mL) is expected to be covered by the range of parasitaemia documented 
in these Australian Phase I IBSM studies:
1. The IBSM study with DSM265 [3] comprised of 8 healthy participants who 
developed asexual blood stage parasitaemia following inoculation with 
P. falciparum parasitised erythrocytes (IBSM model). Among the 7 participants 
who became parasitaemic, the geometric mean parasitaemia upon initial dosing 
with 400 mg DSM265 was 7,851 parasites/mL blood (95% CI, 2,245 to 
27,462 parasites/mL blood). No SAEs were reported. A total of 88 AEs which 
were mostly mild in severity (87.5%) were reported, and the majority were 
deemed related to malaria parasite infection (77.3%). Eleven AEs were moderate 
in severity, while none were classified as severe. The severity of the malaria 
symptoms and signs experienced by the participants during the study was 
generally mild, as assessed by use of a malaria clinical scoring tool. At the time 
the first dose of DSM265 was administered, 4 of the 7 participants who were 
parasitaemic had no malaria symptoms or signs (clinical score, 0), while the 
remaining 3 participants each had a score of 3. The peak malaria clinical score 
recorded during the study was 8; this was recorded for a single participant 
approximately 36 h after administration of the first dose of DSM265.
2. The IBSM study with MMV390048 [13] comprised of 15 healthy volunteers, 
who developed asexual blood stage parasitaemia following inoculation with 
P. falciparum parasitised erythrocytes (IBSM model). As the protocol was calling 
for the full cohort to be inoculated on the same study day, asexual blood stage 
parasitaemia densities prior to receiving any investigational medicinal product 
(IMP) was variable between participants and ranged from 1508 parasites/mL
blood to 95851 parasites/mL blood. Additionally, the malaria clinical score 
associated with the timing of this parasite density in each of these 15 participants, 
included a score of 0/42 (absence of malaria symptoms/signs) in 
10/15 participants, a score of 1/42 in 2/15 participants, a score of 2/42 in 
2/15 participants and a score of 3/42 in 1/15 participants.
3. Due to the mechanism of action of the test drug and evaluation of doses 
associated with suboptimal efficacy, peak asexual blood stage parasitaemia 
densities for these participants was observed after administration of the drug for 
the majority of the participants and ranged from 7946 parasites/mL to 
281387 parasites/mL. Additionally, the malaria clinical score associated with the 
timing of this parasite density in each of these 15 participants, included a score of 
1/42 in 1/15 participants, 2/42 in 6/15 participants, 4/42 in 2/15 participants, 6/42 
in 3/15 participants, 7/42 in 1/15 participants and 8/42 in 2/15 participants.
4. A total of 202 AEs occurred with 176 of these being assessed as 
inoculum-related. Most common AEs reflecting manifestations of blood stage 
malaria infection included: headache (57 cases in 15 participants), myalgia 
(25 cases in 12 participants), malaise (18 cases in 9 participants), reduction in 
lymphocyte count (11 cases in 8 participants), abdominal discomfort (7 cases in 
6 participants), fatigue (10 cases in 5 participants) and arthralgia (7 cases in 
5 participants). Most AEs were of mild severity (145/202). There were 52 AEs of 
moderate severity, which were predominantly signs and symptoms associated 
with malaria infection. There were 5 severe AEs recorded in 3 participants and all 

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considered to be inoculum related (1 participant with severe headache, 
1 participant with transient asymptomatic elevated ALT; elevated AST; and 
decreased lymphocyte count, 1 participant with decreased lymphocyte count). No 
SAEs were reported.
A review of data from 5 completed IBSM-Pf studies that collected both parasitaemia and 
malaria clinical score on first antimalarial treatment day, showed that only a small 
percentage of participants experienced malaria sign/symptoms. The majority had a low 
malaria clinical score of below 3 out of a maximum score of 42 (data on file at MMV and 
QIMR). For the total of 54 participants who were inoculated in these studies, the range of 
parasitaemia at first antimalarial treatment day was 634 to 107981 parasites/mL. At 
treatment initiation, 17 of 54 participants (31.5%) were symptomatic. Of those 
17 participants, 16 (94.1%) had a malaria clinical score ranging from 1-3 and the 
remaining participant had a score of 7. The parasitaemia associated with the clinical 
score of 7 was 31666 parasites/mL whereas the highest parasitaemia observed in this 
analysis (107981 parasites/mL) was observed in an asymptomatic participant
[3,4,6,13,14].
Lastly, it should be noted that from a general safety standpoint, the VIS studies in healthy 
volunteers achieve blood-stage parasitaemia levels that are approximately 1000-fold 
below those observed in patients with acute malaria infection.
Period for In-House Stays in the Clinical Unit
The period of in-house stays proposed for the current study is aimed at maximising 
participant safety and convenience, and ensuring efficient clinical study conduct. 
Participants will be discharged 2 h after inoculation. Experience from previous VIS 
Phase Ib studies with the DVI technique has shown that inoculation is not expected to 
induce inoculum-related AEs or laboratory tests changes (due to incubation period) 
[2,20,27]. Potential tolerability issue is limited to a possible haematoma or vasovagal 
reaction in relation with venepuncture and can be covered by the observation following 
injection.
In line with experience from previous VIS IBSM Phase Ib studies conducted by MMV at 
a Phase I Unit in Australia and the chemoprotection MMV_P218_17_01 study conducted 
in Belgium (EudraCT number: 2018-003004-39), safety will be assessed for both cohorts 
by daily monitoring via phone call from Day 2 until Day 6 and safety, parasitaemia and 
malaria signs and symptoms by daily visits to the clinical unit from Day 7 until Day 9, 
until start of second confinement in the unit on Day 10:
1. Experience with the MMV_P218_17_01 study:
-
Placebo participants developed protocol-defined PCR positivity 
 250 parasites/mL blood) on Day 10 (1/6) or Day 12 (5/6), aligned with a 
geometric mean time to positive parasitaemia of 10.615 (9.918; 11.360) days.
-
Malaria signs and symptoms were reported in 4 (66.7%) participants overall 
and there was a delay between detection of positive parasitaemia and malaria 
sign or symptom onset, with symptoms occurring after PCR positivity and 
starting on Day 12 in 1 out of the 4 participants and on Day 13 for each of the 
remaining 3 participants.

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-
For each of the 4/6 participants, a maximum individual symptom score of one 
(mild) was observed and no moderate or severe sign/symptom was reported. 
-
Total malaria clinical scores for the 4 participants were 2/42, 2/42, 3/42 and 
5/42.
2. Although occurring earlier than Day 10, previous studies [7,10,18] defined PCR 
positivity with a limit of detection between 20 and 30 parasites per mL blood, 
which is lower than our threshold of 250 parasites per mL blood.
Participants will be confined and closely monitored in the unit starting from Day 10 until 
the cohort-specific qPCR parasitaemia and/or malaria clinical scores thresholds for
initiation of registered antimalarial therapy (Riamet®/Malarone®) are reached.
At the time cohort-specific qPCR thresholds (i.e. 5000 parasites/mL blood for Cohort 1 
and 10000 parasites/mL blood for Cohort 2) and/or malaria clinical score (6/42 or more) 
are met, participants will be administered the registered antimalarial therapy. Participants 
will continue to be monitored at the clinical until complete parasite clearance and for a 
minimum of 72 h from initiating antimalarial therapy. Then, if clinically well, 
participants will be discharged and return for an ambulatory EOS visit.
All participants inoculated with PfSPZ-DVI Challenge will commence antimalarial 
therapy no later than Day 24 for both cohorts, regardless if they reach pre-defined 
cohort-specific PCR parasitaemia/malaria clinical score thresholds (i.e., 
5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for Cohort 2 and/or 
DPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQG3DUWLFLSDQWVZKRVWDUWDQWLPDODULDO
therapy on Day 24 will only be discharged from confinement at the end of the EOS visit 
on Day 28.
The Applicant believes there are no further specific measures needed in this proposed 
study due to the use of the registered 3-day regimen of Riamet®/Malarone®.
3.3
END OF STUDY DEFINITION
A participant will be considered to have completed the study if he or she has completed 
all phases of the study including the required EOS visit.
The end of the study is defined as the date of the last EOS visit of the last participant in 
the study.
3.4
SAFETY REVIEW COMMITTEE (SRT)
The study will be overseen by an SRT.
The SRT members will include at least an independent Early Phase and Clinical 
Pharmacology MD expert experienced in malaria volunteer infection studies (SRT chair), 
the Medical Monitor, the Medical/Project Director, the PI and an independent infectious 
disease/malaria expert (or their delegates). The MMV Head of Experimental Medicine 
and Clinical Pharmacology or further internal or external experts such as a clinical lead 
and/or a statistician may be consulted by the SRT as necessary.
The SRT will review safety/tolerability, parasitaemia and malaria signs and symptoms 
data during the study. SRT meetings will be scheduled at one prespecified time (see 
Section 3.4.1) and ad-hoc if deemed necessary. The required data for SRT review will be 

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detailed in a separate study specific SRT Charter and will be sent to the SRT at least 24 h
prior to an SRT meeting. 
The decision of the SRT will be taken in consensus between the members of the SRT. If
consensus cannot be reached, then the most cautious approach will proceed. The SRT 
decision and decision-making will be documented in the SRT meeting minutes, which 
will be provided to the attendees after the meeting.
Safety will be continuously monitored by the Investigators, the Sponsor’s medical 
monitor, and study personnel.
Criteria for discontinuation of dosing in individual participants are listed in Section 8.
The SRT will not review the data from the sentinel groups in Cohort 1 and Cohort 2.
Progression from Subgroup 1 to Subgroup 2 in both Cohorts 1 and 2 will occur upon 
decision of the PI, medical director and medical monitor only (see Section 3.1).
3.4.1
Toxicity Criteria and Progression From Cohort 1 to Cohort 2
An SRT meeting is scheduled to occur at least a week after the conclusion of malaria 
therapy of Cohort 1, prior to commencing Cohort 2. The SRT will review available 
Cohort 1 safety/tolerability and parasitaemia data up to the last dose of Riamet® for all 
inoculated participants in Cohort 1 before enrolment of Cohort 2 and determine whether 
progression to Cohort 2 is indicated. If no safety concerns are identified during SRT 
review, progression to Cohort 2 will take place. Initiation of Cohort 2 will be put on hold 
and further review will be conducted by the SRT, if any of the following toxicity criteria 
are observed in Cohort 1:
-
PfSPZ-DVI Challenge-related or Riamet®-related SAE; or
-
any other critical PfSPZ-DVI Challenge-related finding that may place 
participants at risk within the same cohort or in the next cohort; or
-
two or more Riamet®-related severe (grade 3 or higher) AEs, independent of 
within or not within the same system organ class.
If after data review the SRT confirms that any of the above has been met, including a 
relationship with the PfSPZ and/or Riamet®, progression to Cohort 2 with higher target 
levels of parasitaemia might not be conducted.
3.4.2
Final SRT Review at Study End
The SRT will also meet at the end of the study when data from Cohort 2 are available for 
a final assessment in order to review the study results and share feedback on possible 
safety signals/events. 
Throughout the study, the SRT can be convened at the request of any member should 
they have cause for concern regarding participant safety in relation to the challenge 
agent, where no other cause can be attributed. In the event that it is not possible to 
quickly convene the SRT for a review of the data, or not all the data are available, the 
study will be interrupted or temporarily halted, at the discretion of the PI or Sponsor, at 
any time.

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4. SELECTION OF STUDY POPULATION
Screening for eligible participants will be performed within approximately 4 weeks prior 
to first confinement and up to Day -2.
Approximately 16 participants are planned to be enrolled sequentially in 2 cohorts of up
to 8 participants per cohort. A participant may be enrolled in one cohort only. 
For details on the sample size calculation, please refer to Section 9.2.
4.1
INCLUSION CRITERIA
Participants meeting all of the following criteria are eligible to participate in this study:
1. Informed Consent Form signed voluntarily before any study-related procedure is 
performed, indicating that the participant understands the purpose of and 
procedures required for the study and is willing to participate in the study, 
including administration of registered antimalarial therapy;
2. Male or female, between 18 and 55 years old (extremes included) at screening;
3. Body weight of at least 50 kg and a body mass index (BMI) of 19.0 to 30.0 kg/m2
(extremes included);
4. Good general health without clinically relevant medical illness, physical exam 
findings including vital signs, and laboratory abnormalities (e.g., without liver 
transaminases >1x ULN and according to the clinically acceptable ranges for 
study inclusion laboratory tests in Attachment 4) as determined by the 
Investigator;
5. Willing to adhere to the prohibitions and restrictions specified in this protocol
(see Section 4.3), including willingness to stay confined to the inpatient unit for 
the required duration and willingness to avoid travelling outside of Benelux 
during the study period;
6. Female participants should fulfil one of the following criteria:
a. At least 1 year postmenopausal (amenorrhea >12 months and 
follicle-stimulating hormone [FSH] >30 mIU/mL) prior to screening;
b. Surgically sterile (bilateral oophorectomy, hysterectomy or bilateral 
salpingectomy);
c.
Will use contraceptives as outlined in inclusion criterion 7;
7. Female participants of childbearing potential (excluding females with female 
partners) must agree to the use of a highly effective method of birth control from 
the screening visit until 40 days after the EOS visit at Day 28 (covering a full 
menstrual cycle of 30 days starting after 5 half-lives of last dose of Riamet®);
Note: Highly effective birth control methods include: combined (oestrogen- and 
progestogen-containing) oral/intravaginal/transdermal hormonal contraception associated with 
inhibition of ovulation, progestogen-only oral/injectable/implantable hormonal contraception 
associated with inhibition of ovulation, intrauterine device, intrauterine hormone-releasing 
system, bilateral tubal occlusion, vasectomised partner or sexual abstinence from heterosexual 
intercourse.

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8. Female participant has a negative pregnancy test at screening and upon admission 
in the clinical unit;
1RWH3UHJQDQF\WHVWLQJZLOOFRQVLVWRIVHUXPȕ-KXPDQFKRULRQLFJRQDGRWURSLQȕ-HCG) tests at 
screening and at the EOS visit and a XULQHȕ-HCG tests on Day -1, in all women.
9. Different ways of being reachable 24/7 (e.g., by mobile phone, regular phone or 
electronic mail) during the whole study period.
4.2
EXCLUSION CRITERIA
Participants meeting any of the following criteria are excluded from participation in this
study:
1. Nursing (lactating) women;
2. Participation in any other clinical drug or vaccine study within 30 days (or 
5 half-lives for drugs) preceding the day of PfSPZ-DVI Challenge (whichever is 
longer), or plans to participate in other investigational drug or vaccine research
during the study period;
3. Participants who took standard vaccinations within 3 months before the start of 
the study or are planning to take standard vaccinations during the study period up 
to 8 weeks after PfSPZ-DVI Challenge;
4. Blood product donation to any blood bank during the 8 weeks (whole blood) or 
4 weeks (plasma and platelets) prior to admission in the clinical unit on Day -1;
5. Mean ECG outside normal range and deemed clinically relevant by the 
Investigator. Examples of clinically significant ECG abnormalities for this study 
include:
-
PR-interval >220 ms;
-
QRS-complex >120 ms;
-
Absolute QT greater than >500 ms;
-
QT interval corrected according to Bazett’s formula (QTcB) or QTcF 
>450 ms for male participants, >470 ms for female participants;
-
Pathologic Q wave;
-
Significant ST-T wave changes;
-
Left or right ventricular hypertrophy;
-
Non-sinus rhythm except isolated premature atrial contractions and 
ventricular extrasystole <2 per 10 s ECG lead;
-
Incomplete left bundle branch block, or complete or intermittent right or left 
bundle branch block;
-
Second or third degree A-V heart block.
6. Seropositive human immunodeficiency virus (HIV), hepatitis A 
immunoglobulin M (IgM) antibody, hepatitis B virus (HBV) (hepatitis B surface 
antigen [HBsAg]), hepatitis C virus (HCV) (antibody), hepatitis D antibody, 
hepatitis E IgM antibody, cytomegalovirus (CMV) IgM antibody or Epstein Barr 
Virus (EBV) IgM antibody;

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7. Previous or current diagnosis of hepatitis including but not limited to viral 
hepatitis, auto-immune hepatitis, non-alcoholic steatohepatitis (NASH), 
alpha-1-antitrypsin deficiency, alcoholic liver disease, primary biliary cholangitis 
(PBC), primary sclerosing cholangitis (PSC), hemochromatosis, Wilson disease 
or suspected hepatocellular carcinoma (HCC).
8. History or presence of diagnosed food or known drug allergies (including but not 
limited to allergy to any of the antimalarial medications to be used in the study, 
see Section 5.1), or history of anaphylaxis or other severe allergic reactions;
Note: Participants with seasonal allergies/hay fever, house dust mite or allergy to animals that are 
untreated and asymptomatic at the time of dosing can be enrolled in the study.
9. History of convulsion or severe head trauma, excluding fever convulsion under 
5 years of age;
Note: A medical history of a single febrile convulsion during childhood is not an exclusion criterion.
10. History of serious psychiatric condition that may affect participation in the study 
or preclude compliance with the protocol, including but not limited to past or 
present psychoses, disorders requiring lithium, a history of attempted or planned 
suicide, more than one previous episode of major depression, any previous single 
episode of major depression lasting for or requiring treatment for more than 
6 months, or any episode of major depression during the 5 years preceding 
screening;
Note: The Beck Depression Inventory (Attachment 2) will be used as an objective tool for the 
assessment of depression at screening. In addition to the conditions listed above, participants with 
a score of 20 or more on the Beck Depression Inventory and/or a response of 1, 2 or 3 for item 9 
of this inventory (related to suicidal ideation) will not be eligible for participation. Participants 
with a Beck score of 17 to 19 may be enrolled at the discretion of the Investigator if they do not 
have a history of the psychiatric conditions mentioned in this criterion and their mental state is not 
considered to pose additional risk to the health of the volunteer or to the execution of the study 
and interpretation of the data gathered.
11. A medical, occupational or family problem as a result of alcohol or illicit drug 
abuse during the past 12 months or current alcohol or illicit drug abuse or 
addiction (positive alcohol breath test or positive drug screen for amphetamines, 
barbiturates, benzodiazepines, cannabinoids, cocaine or opiates at screening or 
upon check-in at the clinical unit);
Note: Excessive use of alcohol is defined as an intake of >21 units per week for males and 
>14 units per week for females where one alcohol unit is defined as 10 mL or 8 g of pure alcohol. 
A single unit is equal to one 25-mL (single) measure of whisky (alcohol by volume [ABV] 40%), 
or a third of a pint of beer (190 mL; ABV 5-6%) or half a standard (175 mL) glass of wine (ABV 
12%).
12. Participants are non-smokers or ex-smokers for more than 90 days prior to 
screening, or smoke no more than 5 cigarettes per day. If users of nicotine 
products (i.e., spray, patch, e-cigarette, etc.), they should use the equivalent of no 
more than 5 cigarettes per day. Participants must agree to abstain from smoking 
while in the unit;

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13. Use of any prescription drugs, herbal supplements (e.g., St John’s Wort) or 
over-the-counter medication within 7 days or 5 half-lives (whichever is longer) 
prior to the PfSPZ-DVI Challenge, or an anticipated requirement for the use of 
these during the course of the study (see Section 6.2);
Note: If necessary, the incidental use of non-steroidal anti-inflammatory drugs (NSAIDs) and
paracetamol (2 g/day, 10 g/week) may be acceptable at the Investigator’s discretion and will be 
documented in the eSource system. The use of nutritional supplements during this time that are 
not believed to have the potential to affect participant safety nor the overall results of the study, 
may be permitted on a case-by-case basis by the Investigator.
14. Any surgical or medical condition possibly affecting drug absorption (e.g., 
cholecystectomy, gastrectomy, bowel disease), distribution, metabolism or 
excretion;
15. Personnel (e.g., Investigator, sub-investigator, research assistant, pharmacist, 
study coordinator or anyone mentioned in the delegation log) directly involved in 
the conduct of the study;
16. Any condition that in the opinion of the Investigator would jeopardise the safety 
or rights of a person participating in the study or would render the person unable 
to comply with the protocol;
17. Personal history of malaria;
18. Volunteer has travelled to or lived in a malaria-endemic area within 6 months 
prior to planned study enrolment;
19. Plans to travel to malaria-endemic region during the study period up to last 
follow-up visit;
20. Previous participation in any malaria vaccine or CHMI study/VIS;
21. Falling in moderate or higher risk category for a fatal or non-fatal cardiovascular 
event within 5 years (> 5%) determined by a validated risk estimation system,
e.g., SCORE [21];
22. Use of systemic antibiotics with known antimalarial activity within 5 half-lives of 
PfSPZ-DVI Challenge (e.g., trimethoprim-sulfamethoxazole, doxycycline, 
tetracycline, clindamycin, erythromycin, fluoroquinolones or azithromycin) or an 
anticipated requirement for the use of these during the study period (see 
Section 6.2);
23. Receipt of blood or blood-derived products (including immunoglobulin) within 
3 months prior to screening. Receipt of packed RBCs given for an emergent 
indication in an otherwise healthy person, and not required as ongoing treatment 
is not exclusionary (for example packed RBCs emergently given during an 
elective surgery).
24. Participants who test positive for severe acute respiratory syndrome coronavirus 2 
(SARS-CoV-2).
Note: In case of an out-of-range clinical laboratory test (according to the clinically acceptable 
ranges for study inclusion laboratory tests in Attachment 4), vital sign or ECG value that will
determine a participant’s eligibility, or in case of a positive drug screen, a retest or expert 
evaluation can be requested. Results of any retest must be available prior to inoculation. The result 
of the retest will be considered for participant eligibility at the Investigator’s discretion. 
Participants can be rescreened at the discretion of the Investigator.

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4.3
LIFESTYLE CONSIDERATIONS
No blood donation will be allowed until 24 weeks after the end of the study.
Female participants of childbearing potential (excluding females with female partners) 
must agree to the use of an effective method of contraception from the screening visit 
until 40 days after the EOS visit at Day 28, as outlined in Section 4.1. For details on the 
existing data regarding the reproductive toxicity of the PfSPZ-DVI Challenge, Riamet®
and Malarone®, please see the current Investigator’s Brochure [9] and SmPCs [22] and 
[12].
Information on prohibited therapies can be found in Section 6.
Participants will be confined from Day -1 in the morning until 2 h post inoculation on 
Day 1 and from Day 10 in the morning until the day of parasite clearance and at least 
72 h after initiating antimalarial therapy (estimated to occur on or before Day 19 and on 
or before Day 22 for Cohort 1 and Cohort 2, respectively) or from Day 10 in the morning 
until Day 28 for participants that start antimalarial therapy on Day 24, unless they are 
withdrawn from the study.
Authorisation to leave the clinical unit will be given by the Investigator.
4.3.1
Meals and Dietary Restrictions
1. Safety clinical laboratory blood tests will be performed in a fasted state, i.e., after 
overnight fast for at least 8 h, except for the ones taken during antimalarial 
therapy for troponin T measurements, and except for the ones on Day of first PCR 
SDUDVLWHVP/LQ&RKRUWRUSDUDVLWHVP/LQ&RKRUWDQGSULRUWR
initiating antimalarial therapy, where a non-fasted state is allowed.
2. The first dose of antimalarial drug should be taken as soon as possible according 
to the pre-defined cohort-specific PCR parasitemia/malaria clinical score 
thresholds (i.e., 5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL
EORRGIRU&RKRUWDQGRUDPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQGRURQ
reaching Day 24. Antimalarial drug administration should be immediately 
followed by a meal or drinks rich in fat (e.g., milk). 
3. Participants will abstain from eating or drinking grapefruit or grapefruit-related 
citrus fruits (e.g., Seville oranges, pomelos) from 2 weeks prior to the first 
Riamet® administration and for the whole duration of the study.
4. Participants should not consume poppy-seeds within 24 h before screening and 
before each urine drug screening because this can falsify the results of the opiate 
urine drug test.
4.3.2
Caffeine, Alcohol, and Tobacco
5. Participants will abstain from smoking a tobacco product or marijuana in the unit.
Participants are non-smokers or ex-smokers for more than 90 days prior to 
screening, or smoke no more than 5 cigarettes per day (see Section 4.2).
6. Participants will not be allowed to take any alcohol-containing food or beverages 
and energy drinks containing taurine or glucuronolactone (such as Red Bull) from 

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24 h prior to screening and from 24 h prior to admission to the clinical unit on 
Day 10 until discharge.
4.3.3
Activity
7. Participants will abstain from any strenuous activity (e.g., weight lifting, aerobics, 
football, endurance training sessions) or unaccustomed physical exercise during 
the whole study (from 48 h prior to screening and from 48 h prior to admission to 
the unit on Day -1 until EOS).
4.3.4
Other
8. Transplantation is not permitted until successful antimalarial treatment will be 
given.
4.4
SCREEN FAILURES
Unscheduled visits may be planned to assess, confirm, and follow-up on out-of-range
clinical laboratory test, vital sign, or ECG values that determine a participant’s eligibility, 
or in case of a positive urine drug screen. The result of the retest must be considered for 
participant eligibility and must be available prior to inoculation (Day 1). Findings made 
during unscheduled visits should be reported in the eSource system.
If a participant does not meet all selection criteria (is a screen failure) but at some point 
in the future is expected to meet the eligibility criteria, the participant may be rescreened 
on 1 occasion only. Participants who are rescreened will undergo the informed consent 
process, be assigned a new participant number, and then restart a new screening phase.

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5. STUDY INTERVENTIONS
5.1
STUDY INTERVENTIONS ADMINISTERED
Manufacturing, packaging, and labelling of PfSPZ-DVI Challenge inoculum will be done 
under the responsibility of the biotechnology company Sanaria (USA) [9].
The clinical unit will be responsible for acquiring the registered antimalarial drugs 
artemether-lumefantrine (Riamet®) and atovaquone-proguanil (Malarone®):
-
Participants will be prescribed with Riamet® according to the pre-defined 
cohort-specific PCR parasitaemia/malaria clinical score thresholds (i.e., 
5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for Cohort 2 
and/or a malaria clinical sFRUHIRU&RKRUWs 1 and 2) to ensure parasite 
clearance prior to the end-of-study evaluation. In addition, participants will also 
be administered with Riamet®:
o
if they experience  inoculum-related SAE, irrespective of severity; or
o
if they experience  Common Terminology Criteria for Adverse Events 
(CTCAE) JUDGH3 (severe) AE deemed related to malaria and not 
self-resolved or relieved with concomitant medications; or
o
if the PI or delegate considers it necessary for participant safety. In this 
situation, the PI or delegate will consult with the independent infectious 
disease/malaria expert (Prof. Jean-Pierre Van geertruyden). However, 
antimalarial medication may be administered prior to consultation if
immediate treatment is deemed necessary for participant safety.
-
If an intolerance or contraindication to Riamet® develops, Malarone® will be 
administered. 
All participants inoculated with PfSPZ-DVI Challenge will commence antimalarial 
therapy no later than Day 24 for both cohorts, regardless if they reach pre-defined 
cohort-specific PCR parasitaemia/malaria clinical score thresholds (i.e., 
5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for Cohort 2 and/or 
DPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQG
All participants must consent to receiving antimalarial therapy. Even in the case of 
withdrawal from the study, all participants administered the PfSPZ-DVI Challenge are to 
receive antimalarial therapy as soon as possible, and to have all appropriate visits and 
assessments as required.
The PfSPZ-DVI Challenge agent consists of a strain of P. falciparum NF54 sporozoites 
used for CHMI/VIS studies that is known to be sensitive to the registered antimalarial 
therapy treatments described above.

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Table 2:
Description of Study Interventions
Intervention Name
PfSPZ-DVI
Challenge
artemether-lumefantrine 
(Riamet®)
atovaquone-proguanil 
(Malarone®); only to be 
used if an intolerance or 
contraindication to 
Riamet® develops
Intervention Type
inoculum
drug
Dose Formulation
cryovial
tablet
Unit Dose 
Strength(s)
15000 or 
50000 aseptic, 
cryopreserved 
P. falciparum 
sporozoites
20 mg artemether and 
120 mg lumefantrine
250 mg atovaquone and 
100 mg proguanil 
hydrochloride
Dosage Level
3200 P. falciparum
sporozoites once
6 doses of 4 tablets over 
a period of 3 days at 
approximately 0, 8, 24, 
36, 48 and 60 h
4 tablets as a single daily 
dose for 3 consecutive 
days
Route of 
Administration
intravenously by 
DVI
oral
Meals in Relation 
to Dosing
Not applicable
see Section 4.3.1
Use
challenge agent
antimalarial medication
Sourcing
provided by 
Sanaria (USA)
provided locally by the clinical unit
Packaging and 
Labeling
dispensed into 
screw-cap vials 
containing 
15000 or 
50000 sporozoites 
in a 20 μL-aliquot
light yellow, round 
tablet with ‘NC’
debossed on one side 
and ‘CG’ on the other
round, biconvex, pink 
film-coated tablets 
engraved 'GX CM3' on 
one side
Any deviation from the treatment regimen defined in the protocol must be documented in 
the eSource system.
5.2
PREPARATION/HANDLING/STORAGE/ACCOUNTABILITY
The Investigator (or his/her designee) is responsible for the safe storage of PfSPZ-DVI 
Challenge and all antimalarial drugs assigned to/acquired by the clinical site, in a locked, 
secure storage facility with access limited to those individuals authorized to dispense the 
PfSPZ-DVI Challenge and antimalarial drugs, and maintained within the appropriate 
ranges of temperature. PfSPZ-DVI Challenge and all antimalarial drugs must be stored as 
specified at delivery and in the original packaging. 
Regular temperature logging of the PfSPZ-DVI Challenge and antimalarial drug storage 
room at the clinical site should be performed. In case a deviation in storage conditions 
should occur, the clinical site must not further dispense the affected PfSPZ-DVI
Challenge and antimalarial drugs and notify the Sponsor.

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The Investigator (or his/her designee) is responsible for ensuring that PfSPZ-DVI
Challenge and all antimalarial drugs received at the clinical site are inventoried and 
accounted for throughout the study.
PfSPZ-DVI Challenge and antimalarial drugs should be dispensed under the supervision 
of the Investigator, a qualified member of the clinical staff, or by a hospital/clinic 
pharmacist. The Investigator must maintain accurate records demonstrating date and 
amount of PfSPZ-DVI Challenge and drugs supplied to whom and by who. PfSPZ-DVI
Challenge and antimalarial drugs will be supplied only to participants participating in the 
study.
The Sponsor’s designated site monitor will periodically check the supplies of PfSPZ-DVI
Challenge and antimalarial drugs held by the Investigator or pharmacist to ensure 
accountability and appropriate storage conditions of all products used.
Unused products must be available for verification by the site monitor during on-site 
monitoring visits. 
After the last visit of the last participant in the study (LPLV), any unused PfSPZ-DVI 
Challenge will be returned to Sanaria. Any unused antimalarial medication will be
destroyed at the clinical site (via Emergency waste and chemical services [EWACS])
after Sponsor’s approval (in this case a certificate of destruction will be provided and 
filed in the Trial Master File [TMF])
Hazardous materials such as used ampules, needles, syringes and vials containing 
hazardous liquids, should be disposed of immediately in a safe manner and therefore will 
not be retained for drug accountability purposes.
5.2.1
PfSPZ-DVI Challenge
In brief, the manufacturing process includes the production, under traditional 
environmental condition, of eggs from a colony of A. stephensi mosquitos housed in a 
controlled environmental chamber. Surface disinfection of the eggs is performed by 
exposure to chemical agents in a Class II biosafety cabinet (BSC). Thereafter, all 
materials and product are handled using aseptic methods to ensure that contaminating 
microorganisms are not introduced to, and carried through, the process. 
Surface-disinfected eggs are inoculated into sterile, vented flasks containing aseptic 
growth medium. The eggs hatch and develop into pupae, which are transferred to an 
adult mosquito container from which the adult mosquitoes emerge. 
These adult mosquitos, which have been raised under aseptic conditions, are fed 
P. falciparum gametocyte-infected blood in a BSC in a high-security insectary in 
Rockville, Maryland, USA. The P. falciparum gametocyte-infected blood is produced 
from cultures of the P. falciparum strain NF54, derived from a master cell bank of the 
well characterised P. falciparum strain NF 54. Infected adult mosquitos are maintained 
under aseptic conditions until P. falciparum sporozoites migrate to the salivary glands. 
The salivary glands from the P. falciparum sporozoite infected mosquitos are removed 
by hand dissection and then triturated to release the P. falciparum sporozoites. 
The sporozoites are purified, counted, and at a specified concentration, cryopreserved. 
Cryopreservation commences with the addition of cryoprotective additives to the purified 
sporozoites to produce the PfSPZ-DVI Challenge agent.

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The diluent for PfSPZ-DVI Challenge is composed of phosphate buffered saline (PBS) 
and human serum albumin (HSA). Vials of PBS and HSA will be shipped to the clinical 
unit, where diluent composed of PBS and HSA is prepared according to a local Standard 
Operational Procedure, as described in the Laboratory Manual of Sanaria (Storage, 
Preparation and Administration of PfSPZ-DVI Challenge). 
PBS that is manufactured in compliance with Good Manufacturing Practice (GMP) and
according to upstream processing specifications is purchased by Sanaria. Every lot of 
PBS is supplied with a batch certification that is reviewed and approved upon receipt of 
Sanaria. The PBS is stored at ambient temperature in a controlled room. 
HSA (25%), approved for parenteral, intravenous administration to humans, is purchased 
by Sanaria. Every lot of HSA is supplied with a Certificate of Analysis that is reviewed 
and approved upon receipt of Sanaria. HSA vials are stored at ambient temperature in a
controlled room.
The PfSPZ-DVI Challenge agent is dispensed into screw-cap vials containing 15000 or 
50000 sporozoites in a 20 μL aliquot. The PfSPZ-DVI Challenge is stored in liquid 
nitrogen vapour phase at -140°C to -196°C until it is shipped to the clinical unit. 
Shipment is in compliance with U.S. Food and Drug Administration (FDA), U.S. 
Department of Transportation and United Nations transport guidelines for shipping bio-
hazardous materials on dry ice and liquid or vapour phase nitrogen.
Transfer of PfSPZ-DVI Challenge from its storage site to the clinical unit will follow 
local Standard Operational Procedure, as described in the Laboratory Manual of Sanaria. 
At the clinical unit, the liquid nitrogen vapour phase container will be monitored. Receipt 
of the PfSPZ-DVI Challenge will be documented on a tracking log by study staff.
Immediately prior to use, the PfSPZ-DVI Challenge in cryovials will be thawed 
individually by partial submersion of the vials for 30 s in a 37°C ± 1°C water bath. 
Designated, trained study staff will then prepare, dilute (if necessary) and dispense the 
PfSPZ-DVI Challenge to clinical staff at the clinical unit according to local standard 
Operational Procedures and as described in the Laboratory Manual of Sanaria.
The PfSPZ-DVI Challenge (3200 P. falciparum sporozoites per participant) will be 
administered intravenously by DVI. The study staff administering the PfSPZ-DVI
Challenge will wear gloves and eye protection. Advanced life support drugs and 
resuscitation equipment will be immediately available in the event of any participants
experiencing an anaphylactic reaction to the challenge.
5.2.2
Registered Antimalarial Therapy
Riamet® and Malarone® must be stored at ambient conditions (15-30°C or 59-86°F),
should not be exposed to freezing temperatures, and should be protected from light
during storage at the clinical site.
Procedures for handling and storage are detailed on the study-specific labels [22,12].

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5.3
RANDOMISATION AND BLINDING
Participants are not randomised. As participants are confirmed to be eligible for the 
study, they will be assigned a single unique identifier across the study and will be 
enrolled sequentially in 1 of the 2 cohorts.
All participants will be inoculated with PfSPZ-DVI Challenge and receive antimalarial 
therapy. PfSPZ-DVI Challenge will be packaged and labelled by Sanaria in line with 
local labelling regulations. The antimalarial drugs (Riamet® and Malarone®) will be 
labelled by the clinical unit pharmacy with an appropriate study label in line with local 
labelling regulations. The pharmacist or appropriate qualified member of the study staff, 
assigned by the PI, will prepare the PfSPZ-DVI Challenge and antimalarial drugs 
identifying the products with the correct participant unique identifier according to the 
applicable procedures.
As this is an open-label study, blinding is not applicable.
5.4
DOSE MODIFICATION
No dose modification is allowed.
5.5
TREATMENT COMPLIANCE
To ensure treatment compliance, PfSPZ-DVI Challenge and Riamet®/Malarone® will be 
administered/intakes will be supervised by the Investigator or his/her designee.
Any deviation from the treatment regimen defined in the protocol must be documented in 
the eSource system.

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6. PRIOR AND CONCOMITANT THERAPY
All therapies (prescriptions and over-the-counter medications, including herbal 
preparations/treatments) other than the antimalarial medication administered from the 
time of informed consent until the last study visit must be recorded in the eSource system 
(name of the drug, dosage, route and dates of administration).
Medications taken during the 28 days prior to PfSPZ-DVI Challenge will be recorded in 
the eSource system as previous medications. Medications taken after this time will be 
recorded as concomitant medications.
Female participants of childbearing potential (excluding females with female partners) 
must agree to the use of an effective method of contraception throughout the study, as 
outlined in Section 4.1. The use of oral, injectable and implantable hormonal 
contraceptives is to be recorded in the eSource system.
6.1
PERMITTED CONCOMITANT THERAPIES
Participants will abstain from using any medications (prescription or over-the-counter) or 
herbal remedies from 7 days or 5 half-lives (whichever is longer) prior to PfSPZ-DVI 
Challenge, as described in the exclusion criteria of this protocol (see Section 4.2). If 
necessary, the incidental use of NSAIDs and paracetamol (2 g/day, 10 g/week), and the 
use of nutritional supplements may be acceptable at the Investigator’s discretion.
Except for the antimalarial medication and medication considered essential to treat AEs, 
all medications (prescription and non-prescription), herbal remedies and nutritional 
supplements should be avoided within 7 days or 5 half-lives (whichever is longer) prior 
to the PfSPZ-DVI Challenge until after the EOS visit on Day 28. Incidental and limited 
use of medications not believed to affect participant safety, nor the overall results of the 
study may be permitted on a case-by-case basis, following approval by the Sponsor in 
consultation with the Investigator, as described in the exclusion criteria of this protocol 
(see Section 4.2).
Due to possible transient asymptomatic ALT and AST elevations after malaria 
inoculation, ibuprofen (at doses of up to 1.8 g/day) is the preferred treatment over 
paracetamol for possible emergent malaria symptoms. In the case paracetamol is used, 
the recommended dose should not exceed 2 g/day.
6.2
PROHIBITED CONCOMITANT THERAPIES
The following medications are not allowed during the study and should be washed-out 
within 5 half-lives of PfSPZ-DVI Challenge:
-
Drugs with known antimalarial activity (trimethoprim-sulfamethoxazole, 
tetracycline, doxycycline, erythromycin, clarithromycin, azithromycin, 
clindamycin, rifampicin, or newer quinolones, benzodiazepines, flunarizine, 
fluoxetine, methotrexate, chloroquine and hydroxychloroquine);
-
Any drug susceptible to interaction with breast cancer resistance proteins (BCRP)
(cyclosporin, elacridar, eltrombopag, gefitinib).

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7.
ASSESSMENTS
7.1
TIMING OF ASSESSMENTS
An overview of the timing of PfSPZ-DVI Challenge, treatment and assessments is given 
in the Time and Events Schedule.
If assessments are planned at the same time point in the study, the order of assessments 
should be as follows:
1. 12-lead ECG recordings;
2. vital signs measurements;
3. blood sampling;
4. other assessments (except for urinalysis assessments, morning void allowed).
The following time-related windows for parasitaemia assessments will be acceptable 
based on logistical and operational considerations:
-
± 60 min on Days 7, 8 and 9;
-
± 10 min during confinement;
-
± 1 day at the EOS visit.
The following time-related windows for blood pressure, pulse rate, ECG, haematology, 
blood chemistry and urinalysis assessments will be acceptable based on logistical and 
operational considerations:
-
pre-inoculation observations within 3 h prior to inoculation;
-
a 5% deviation from theoretical post inoculation times will be allowed for all 
other post inoculation assessments.
All assessments scheduled for a particular time point should be performed within these 
windows.
Any deviations from the above-mentioned window periods will be documented as 
protocol non-compliances and will be evaluated case-by-case for significance at the data 
review meeting prior to database lock.
Adverse events and the intake of concomitant medication(s) will be monitored 
continuously from informed consent until the last study-related activity.
7.1.1
Screening Period
Screening Visit
Screening for eligible and consenting participants will be performed within 
approximately 4 weeks prior to first confinement.
Participants will be given a full explanation of the nature of the study and written 
informed consent (approved by the local ethics committee) will be obtained according to 
local requirements before any study-related assessment will be carried out.

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At screening, participants will be asked to attend the clinical unit to have assessments 
performed as indicated in the Time and Events Schedule.
All results from the screening procedure needed to evaluate eligibility, including the 
clinical laboratory results, must be available before PfSPZ-DVI Challenge on Day 1. Any 
abnormal assessment at the screening visit will be assessed according to its clinical 
relevance, and if found relevant, the participant will not be included in the study (see also 
Section 4.4).
Up until Day -2, testing for SARS-CoV-2 using real-time reverse transcription PCR 
(rRT-PCR) on nasopharyngeal swabs will also be performed for participants who 
completed the rest of the screening procedure and are still eligible to be enrolled in the 
study. The rRT-PCR test will be performed by ZNA Middelheim or by SGS Clinical 
Pharmacology Unit, depending on the availability of appropriate equipment.
Unscheduled visits may be planned to assess, confirm and follow-up on out-of-range 
clinical laboratory test, vital sign or ECG values that determine a participant’s eligibility, 
or in case of a positive urine drug screen. The result of the retest will be considered for 
participant eligibility. Findings made during unscheduled visits should be reported in the 
eSource system.
Pre-treatment Day -1
Participants will be admitted to the clinical unit in the morning of Day -1 (the day before 
PfSPZ-DVI Challenge) and participants will remain at the clinical unit until 2 h post 
inoculation on Day 1.
Eligibility of the participants will be confirmed and assessments will be performed as 
indicated in the Time and Events Schedule. All assessments must be completed and 
safety assessment outcomes must be available before PfSPZ-DVI Challenge on Day 1.
7.1.2
Treatment Period
Day 1
On Day 1, assessments begin as of 3 h before PfSPZ-DVI Challenge (except for 
urinalysis assessments, morning void allowed) and will be performed as indicated in the 
Time and Events Schedule. All assessments must be completed and safety assessment 
outcomes must be available before inoculation with approximately 3200 P. falciparum
sporozoites (NF54 strain) by DVI. After assessments are performed as indicated in the 
Time and Events Schedule, participants will be discharged 2 h post inoculation.
Day 2 Until Day 6
Participants will be monitored daily via phone call from Day 2 until Day 6 to solicit any 
AEs.
Day 7 Until Day 9
Participants will come to the clinical unit daily from Day 7 until Day 9 and the presence 
of parasites will be assessed once daily by qPCR.
Before confinement to the study site (on Day 10), participants will be tested again for 
SARS-CoV-2 using rRT-PCR on nasopharyngeal swabs. If a participant tests positive for 
SARS-CoV-2, he or she will be withdrawn from the study and will be administered 

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registered antimalarial therapy (i.e., Riamet®) immediately. The SARS-CoV-2 infection 
will be managed further according to national laws.
Day 10 Until Day of Pre-Defined Cohort-Specific Thresholds (Including Day of First 
3&53DUDVLWHVP/)
Participants will be confined to the clinical unit from Day 10 in the morning and the 
presence of parasites will be assessed twice daily by qPCR until Day of first PCR 
 parasites/mL blood IRU&RKRUWRUXQWLO'D\RIILUVW3&5 parasites/mL 
blood for Cohort 2.
Assessments will be performed as indicated in the Time and Events Schedule.
Day of Pre-Defined Cohort-Specific Thresholds Until K$IWHU,QLWLDWLQJ
Antimalarial Therapy
Participants will be administered registered antimalarial therapy, i.e., Riamet®, as soon as 
possible when the pre-defined cohort-specific PCR parasitaemia/malaria clinical score 
thresholds (i.e., 5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for 
&RKRUWDQGRUDPDODULDFOLQLFDOVFRUHIRU&RKRUWV 1 and 2) are reached or at the 
Investigator’s discretion (see Section 5.1).
All participants inoculated with PfSPZ-DVI Challenge will commence antimalarial 
therapy no later than Day 24 for both cohorts, regardless if they reach pre-defined 
cohort-specific PCR parasitaemia/malaria clinical score thresholds (i.e., 
5000 parasites/mL blood for Cohort 1 and 10000 parasites/mL blood for Cohort 2 and/or 
DPDODULDFOLQLFDOVFRUHIRU&RKRUWVDQG
Antimalarial therapy will be further administered and monitored. qPCR assessments of 
parasitaemia will be carried out at multiple time points (2, 6, 8, 12, 16, 24, 36, 48 and 
72 h) following initiation of Riamet®.
Assessments will be performed as indicated in the Time and Events Schedule.
'D\RI3DUDVLWH&OHDUDQFH$QGK$IWHU,QLWLDWLQJ$QWLPDODULDO7KHUDS\
Upon parasite clearance and at least 72 h after initiating antimalarial therapy (estimated 
to occur on or before Day 19 and on or before Day 22 for Cohort 1 and Cohort 2,
respectively), and if clinically well, participants will be discharged from the clinical unit.
Participants who start antimalarial therapy on Day 24 will only be discharged from 
confinement at the end of the EOS visit on Day 28.
Assessments will be performed as indicated in the Time and Events Schedule.
7.1.3
Follow-up Period
After discharge from the clinical unit, or after confinement until the EOS on Day 28 in 
case antimalarial therapy was initiated on Day 24, all participants will be followed up for 
safety assessments, clinical evaluation and malaria qPCR in the clinical unit at the EOS 
visit on Day 28.
All participants will be assessed for parasitaemia and will be asked non-leading questions 
to determine the occurrence of any AEs at the EOS visit on Day 28. In order to provide 
some flexibility for the participants regarding the site visit and to maintain the integrity 

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of the study design, a time window of ±1 day is permitted, in case of time conflict or 
unforeseen circumstances. 
Assessments will be performed as indicated in the Time and Events Schedule.
7.1.4
Unscheduled Visits
Unscheduled visits can be planned for instance:
-
to obtain additional information to ensure safety to the participant. Additional 
blood and urine samples may be taken at the discretion of the Investigator;
-
to assess, confirm and follow-up on out-of-range clinical laboratory test, vital sign 
or ECG values that will determine a participant’s eligibility, or in case of a 
positive drug screen. The result of the retest will be considered for participant
eligibility.
Findings made during unscheduled visits should be reported in the eSource system.
7.2
PHARMACODYNAMIC ASSESSMENTS
For an overview of pharmacodynamic endpoints, see Section 2.
7.2.1
Parasitaemia
A blood sample will be collected via direct venepuncture from each participant at time 
points where parasitological assessments are scheduled (Time and Events Schedule). The 
assessment of malaria parasitology by qPCR will be as follows: varATS multigenic 
family targeted qPCR assay of parasite load will be performed in accordance with the 
centre of excellence for tropical medicine (ITM) standard operating procedure and the 
Laboratory Manual. Given the high sensitivity of qPCR, this method will be used to 
confirm parasite clearance after definitive antimalarial therapy for all participants. The 
results of the PCR at the ITM will be available in approximately 6 h from arrival of 
samples at the ITM. A participant will be considered cured following completion of the 
course of antimalarial therapy and after qPCR indicates 0 parasites per mL blood.
7.3
SAFETY ASSESSMENTS
The safety assessment in this study will be based on AEs, the malaria clinical score, 
clinical laboratory tests, vital signs, ECG, physical examination and Beck Depression 
Inventory (screening only) as described in the following sections.
7.3.1
Adverse Events
Adverse events will be monitored continuously from informed consent until the last 
study-related activity. At regular intervals during the study, participants will be asked 
non-leading questions to determine the occurrence of any AEs. All AEs reported 
spontaneously during the course of the study will be recorded as well.
For detailed definitions and reporting procedures of AEs, see Section 10.
Close monitoring of expected signs and symptoms associated with malaria infection 
(listed in Attachment 1) form part of the safety evaluation of the study and will be used 
as part of the decision criteria to administer registered antimalarial therapy (see 

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Section 5.1 and Section 7.3.2). These events will be classified as AEs, and may also be 
further sub-classified as inoculum-related AEs:
An inoculum-related AE is a sign or symptom associated with malaria infection, i.e., 
confirmed by a protocol-defined PCR positivity (defined for the purpose of this study as 
a qPCR outcome  parasites per mL blood) at
-
the onset of the event; or
-
if the event started within 1 day of parasite clearance (defined as a qPCR value of 
0 parasites per mL blood after initiating antimalarial therapy) or 
laboratory-defined PCR positivity (a qPCR outcome of 50-249 parasites per mL 
blood).
If the presence of P. falciparum malaria is confirmed (see above), then these AEs will be 
further sub-classified as inoculum-related AEs, following the guidelines on AE reporting, 
i.e., followed-up until resolution with the same causality assessment (see Section 10), and 
reported as such in the final clinical study report.
If the PCR results for P. falciparum are between 0 and <250 parasites per mL blood at 
the time of the onset of the event and the day before, usual AE/SAE reporting procedures 
and criteria will apply (see Section 10), but the event will not be classified as an 
inoculum-related AE.
Final classification of signs and symptoms as AE and inoculum-related AE, or as AE 
only, will occur after PCR results are available.
7.3.2
Malaria Clinical Score
The malaria clinical score, which assesses malaria signs and symptoms (see 
Attachment 1) at the time points indicated in the Time and Events Schedule, will be part 
of the close monitoring for known/identified risks related to PfSPZ-DVI 
Challenge/malaria. The malaria clinical score has been widely used in the conduct of 
IBSM CHMI/VIS studies in Australia and also implemented in the recently conducted 
PfSPZ-DVI Chemoprotection Challenge study with P218 (MMV_P218_17_01 study 
[EudraCT number: 2018-003004-39]) at the SGS Clinical Pharmacology Unit in 
Antwerp, Belgium [2]. This quantitative tool will be used as part of the decision criteria 
to administer registered antimalarial therapy along with the level of parasitaemia (see 
Section 5.1).
The following 14 signs/symptoms frequently associated with malaria will be graded 
using a 4-point scale (absent: 0; mild: 1; moderate: 2; severe: 3) and summed to generate 
a total malaria clinical score (maximum score possible is 42): headache, myalgia (muscle 
ache), arthralgia (joint ache), fatigue/lethargy, malaise (general discomfort/uneasiness), 
chills/shivering/rigors, sweating/hot spells, anorexia, nausea, vomiting, abdominal 
discomfort, fever, tachycardia and hypotension. To determine severity of the 
14 signs/symptoms we use the CTCAE grading scale grade 1 - 5. Mild (1) equates to 
CTCAE grade 1, Moderate (2) equates to CTCAE grade 2 and Severe (3) equates to 
CTCAE grade 3 or above. Individual scores for each symptom as well as the total score 
will be recorded.
For the grading of fever, the vital sign parameter body temperature (sublingual) will be 
assessed (see Section 7.3.4).

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7.3.3
Clinical Laboratory Tests
Blood samples will be collected by venepuncture or via indwelling cannula at the time 
points indicated in the Time and Events Schedule. Biochemistry and haematology testing
will be performed on these samples, as well as viral serology testing (hepatitis A IgM 
antibody, HbsAg, anti-HCV antibody, hepatitis D antibody [only in subjects positive for 
HbsAg], hepatitis E IgM antibody, CMV IgM antibody, EBV IgM antibody and HIV) on 
the sample from screening. In all female participantVDOVRVHUXPȕ-HCG assessments at 
screening and at the EOS visit and a XULQHȕ-HCG assessment on Day-1 will be 
performed. FSH will be measured at screening in all women.
All blood samples for safety assessments should be taken in a fasted state (after overnight 
fast for at least 8 h for unbiased glucose determination), except for the ones taken during 
antimalarial therapy for troponin T measurements, and except for the one on Day of first 
PCR SDUDVLWHVP/LQ&RKRUWRUSDUDVLWHVP/LQ&RKRUWDQGSULRUWR
initiating antimalarial therapy, where a non-fasted state is allowed.
Standard laboratory tests will be performed by ZNA Middelheim.
The following biochemistry and haematology tests will be performed on the safety blood 
samples: 
-
Liver biochemistry: albumin, AST, ALT, alkaline phosphatase (ALP), gamma 
glutamylaminotransferase (GGT), total and direct bilirubin and total serum 
proteins;
-
Biochemistry other than liver: sodium, potassium, chloride, bicarbonate, urate, 
inorganic phosphate, creatinine, estimated glomerular filtration rate (eGFR), 
glucose, lactate dehydrogenase (LDH), blood urea nitrogen (BUN) and creatine 
phosphokinase (CPK);
-
High sensitive troponin T and;
-
C-reactive protein (CRP);
-
Haematology: haemoglobin, haematocrit, RBC count, mean corpuscular 
haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), 
mean corpuscular volume (MCV), white blood cell (WBC) count, platelet count, 
reticulocytes, neutrophils, eosinophils, basophils, lymphocytes and monocytes;
-
Coagulation: prothrombin time (PT) and activated partial thromboplastin time 
(aPTT) (at screening and on Day -1 only), and international normalized ratio 
(INR).
A midstream urine sample will be collected for urinalysis by dipstick for glucose, 
protein, nitrite, pH and occult blood at the time points for biochemistry other than liver 
indicated in the Time and Events Schedule. Microscopic examination for WBC, RBC 
and casts will be performed.
A urine drug screen (amphetamines, barbiturates, benzodiazepines, cannabinoids, 
cocaine and opiates) and an alcohol breath test will be performed at the time points 
indicated in the Time and Events Schedule.
The Investigator must review the laboratory report, document this review and record any 
change occurring during the study he/she considers to be clinically relevant in the 

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eSource system. Laboratory values outside the normal range will be flagged and their 
clinical relevance will be assessed by the Investigator.
7.3.4
Vital Signs
Vital sign parameters will be assessed after at least 10 min in supine position at the time 
points indicated in the Time and Events Schedule. The vital sign parameters that will be 
assessed are supine systolic and diastolic blood pressure (SBP and DBP, respectively)
and pulse rate. Body temperature (sublingual) will also be assessed, and this whenever 
the malaria clinical score will be evaluated (see Section 7.3.2). Orthostatic changes to BP 
and pulse rate will also be assessed at screening: participants will be requested to stand 
after completion of the supine measurements and blood pressure and pulse rate will be 
recorded after at least 2 min in the standing position.
These parameters will be measured using a completely automated device consisting of an 
inflatable cuff and an oscillatory detection system. All values will be registered on a 
built-in recorder so that measurements are observer-independent.
Any change from baseline in vital sign values occurring during the study that is 
considered to be clinically relevant by the Investigator should be recorded in the eSource 
system.
7.3.5
Electrocardiogram
Twelve-lead ECG recordings will be recorded in triplicate after at least 10 min in supine 
position at the time points indicated in the Time and Events Schedule.
Paper speed will be 25 mm/s, so that the different ECG intervals can be measured 
manually.
The interpretations of the ECGs will be performed by the Investigator or his/her designee 
at the clinical unit. Any change from baseline ECG occurring during the study that is 
considered to be clinically relevant by the Investigator should be recorded in the eSource 
system.
7.3.6
Physical Examination
Physical examination will be performed at the time points indicated in the Time and 
Events Schedule.
Height is to be measured barefoot and at screening only. Body weight to be measured as 
indicated in the Time and Events Schedule. To obtain the actual body weight, 
participants must be weighed lightly clothed.
Any change in physical examination occurring during the study that is considered to be 
clinically relevant by the Investigator should be recorded in the eSource system.
Full physical examination will be conducted at screening, on Day -1, within ±24 h after 
ILUVW3&5SDUDVLWHVP/EORRGDQGDWWKHEOS visit on Day 28. Targeted, 
symptom-driven physical examination will be performed in case malaria clinical 
score >6 on Day 10 (upon confinement to the clinical unit), within 24 h after first PCR 
&RKRUWRU&RKRUW parasites/mL blood and at pre-discharge from 
clinical unit; and will be focused on changes since the previous examination, but will 

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always include at least: chest/respiratory, general appearance, heart/cardiovascular, 
abdomen, neurological and skin/mucous membranes examination.
7.3.7
Beck Depression Inventory
The Beck Depression Inventory is performed at screening only (see Attachment 2).
The questionnaire is scored by the participant. The inventory completed by the 
participant will be reviewed/checked for completeness and the total score calculated by 
the study personnel.
7.4
PARASITE TRANSCRIPTOMICS
Different environmental conditions that occur during a human infection (such as the 
presence of drugs) can select for parasites with different expression patterns [16,17]. We 
aim to evaluate the effect of in-host factors on parasite transcriptional/epigenetic 
responses during experimental infections. 
To this aim, blood samples will be collected via direct venepuncture at the time points 
indicated in the Time and Events Schedule. Sample aliquots will be stored in Trizol for 
RNA preservation and cryopreserved for parasite culture in case higher parasite densities 
are needed to increase starting material for transcriptomic analysis of parasite isolates.
Protocols for transcriptomic analysis of parasite isolates have been validated and are 
currently in use at the Malariology Unit at the ITM.
7.5
TREATMENT OF OVERDOSE
In this study, all dosing will be supervised by study personnel and consequently the risk 
of overdose will be minimal.
7.6
TOTAL VOLUME OF BLOOD SAMPLING
The total volume of blood drawn per the Time and Events Schedule from each 
participant will not exceed the maximum allowable volume of approximately 460 mL.
7.7
APPROPRIATENESS OF MEASUREMENTS
The assessments which will be made in this study are standard, and are generally 
recognized as reliable, accurate and relevant.

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8. PARTICIPANT WITHDRAWAL FROM THE STUDY
8.1
WITHDRAWAL OF PARTICIPANTS FROM STUDY
Participants may be withdrawn from the study following inoculation with PfSPZ-DVI 
Challenge in the event of:
-
a severe AE or a SAE considered related to the PfSPZ-DVI Challenge.
Participants may be withdrawn from the study following inoculation with PfSPZ-DVI 
Challenge and administration of Riamet® in the event of:
-
a severe AE or a SAE considered related to either Riamet® or the PfSPZ-DVI 
Challenge.
Participants must be withdrawn from the study in the event of:
-
withdrawal of consent;
-
safety reasons, it being in the best interest of the participant that he/she be 
withdrawn, in the Investigator’s opinion or in the opinion of the SRT (see 
Section 3.4);
-
a positive pregnancy test (the participant or, in case of a male participant, his 
female partner), or if the participant/partner is non-compliant with the 
contraception requirements (see Section 4.1);
-
a positive SARS-CoV-2 test (see Section 7.1.2);
-
development of a medical condition that requires concomitant treatment with a 
prohibited therapy (see Section 6.2);
-
failure of the participant to comply with the protocol requirements or to cooperate 
with the Investigator resulting in a significant risk to the participant’s safety.
In the event a participant has to discontinue Riamet®, the monitor and Sponsor should be 
informed: in case of withdrawal due to an SAE (for details on AE reporting see 
Section 10), the Sponsor should be notified within 24 h; in case of withdrawal for other 
reasons, the Sponsor should be notified within 2 days from the event.
If there is a medical reason for withdrawal, the participant will remain under the 
supervision of the Investigator until satisfactory health has returned.
All participants must consent to receiving antimalarial therapy if inoculated with the 
PfSPZ-DVI Challenge agent, i.e., the registered 3-day Riamet® regimen approved for 
treatment of uncomplicated malaria. Even in the case of withdrawal from the study, all 
participants administered the PfSPZ-DVI Challenge are to receive antimalarial therapy as 
soon as possible (see Section 5.1), and to have all appropriate visits and assessments as 
required (see Time and Events Schedule). Participants will be followed up for safety 
assessments, clinical evaluation and malaria qPCR in the clinical unit at the EOS visit on 
Day 28. Participants who fail to return for visits will be traced as described in 
Section 8.2.
Participants who have to discontinue Riamet® prior to completion of the scheduled study 
procedures for safety reasons/due to intolerance must complete antimalarial therapy with 
Malarone® and should be invited to complete the assessments as much as possible: as 

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long as the participant consents, all relevant assessments of the day on which the 
participant discontinued should be completed, at least those related to safety, and the 
participant should come for a safety follow-up visit 4 weeks after the last administration 
of antimalarial drug. In the event of discontinuation due to an AE, the appropriate 
follow-up will be done.
Participants have the right to withdraw from the study at any time for any reason, 
including personal reasons. A participant can withdraw without giving a reason. This will 
not affect his/her future care. The Investigator should however try to find out why a 
participant withdraws from the study and document the reason for withdrawal in the 
source documents and in the eSource system.
If the participant withdraws consent for disclosure of future information, the Sponsor 
may retain and continue to use any data collected before such a withdrawal of consent.
Participants who are withdrawn from the study for reasons other than safety may be 
replaced. This decision will be made based upon discussion and mutual agreement 
PfSPZ-DVI Challenge and antimalarial drugs assigned to a participant who withdraws 
must not be assigned to another participant.
8.2
LOST TO FOLLOW-UP
Before a participant is considered lost to follow-up, every reasonable effort must be made 
by the study site personnel to contact the participant, if only to determine the reason for 
discontinuation. The measures taken to follow up must be documented.
The following measures will be taken to assure traceability of the participants:
-
Following daily contact from Day 2 to 9 and scheduled, in-house confinement 
within the clinical unit, participants will be followed up for safety assessments, 
clinical evaluation and malaria qPCR in the clinical unit at the EOS visit on 
Day 28.
-
All participants will commit to remaining contactable 24/7 throughout the study 
and this will be recorded on the Informed Consent Form (ICF).
-
Participants will be required to provide current contact details (two telephone 
numbers including a mobile number and a responsible adult as an emergency 
contact) and a relevant email address.
-
Should a participant not be contactable by the unit, all alternative methods 
possible (e.g., sending someone to home address of participant) will be used to 
locate and communicate with the participant.
-
Should a participant still be uncontactable the situation will be discussed with the 
Sponsor and escalated as appropriate including alerting the regulatory authorities 
to a deviation from protocol.
Should the participant remain unreachable, he/she will be considered to have 
discontinued the study.

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9. STATISTICAL METHODS
9.1
STATISTICAL ANALYSIS
All statistical analysis will be performed by SGS Life Sciences, under the supervision 
and responsibility of the Sponsor, using SAS® (SAS Institute Inc., Cary, NC, USA; 
version 9.4 or higher) software for statistical computations. 
All statistical methods shall be detailed in a Statistical Analysis Plan (SAP) that will be 
finalised before database lock.
The final analysis will be performed once all participants have completed the follow-up
visit or have discontinued earlier. 
Data from all participants enrolled and inoculated with PfSPZ-DVI Challenge will be 
included in the data analysis. The following analysis populations will be defined:
-
Inoculation Set: all participants who were inoculated with PfSPZ-DVI Challenge;
-
PD Analysis Set: all inoculated participants with at least one available PD data 
who received all Riamet® doses and who experienced no protocol deviations with 
relevant impact on PD data;
-
Safety Analysis Set: all inoculated participants who received any treatment.
Unless specified otherwise, the Inoculation Set will be used for analysis of 
demographics, the PD analysis set will be used for pharmacodynamic statistical analysis 
and the Safety Analysis Set will be used for safety/tolerability analysis.
9.1.1
Initial Characteristics Data of the Participant Sample
For all participants who receive PfSPZ-DVI Challenge, descriptive statistics will be 
provided for demographic (e.g., age, height, weight, BMI, race, gender) and other initial 
participant characteristics (alcohol and drug screening tests, pregnancy test, orthostatic 
changes to blood pressure and pulse rate, serology, medical and social history, 
concomitant diseases, Beck Depression Inventory). Standard descriptive statistics for 
continuous variables are the number of participants (N), mean, standard deviation (SD), 
median, minimum and maximum values. The standard descriptive statistics for 
categorical variables are the number of participants in the category and the proportion 
expressed as a percentage.
Prior and concomitant medications will be coded using the WHO_DRUG Dictionary.

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9.1.2
Pharmacodynamic Data
Parasitaemia
Primary endpoints:
To characterise key stages in the parasite growth:
-
first PCR positivity;
-
threshold of 5000 parasites/mL for Cohort 1 and threshold of 10000 parasites/mL 
for Cohort 2; and
-
parasitaemia levels at drug administration.
This will be done by calculating:
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of time to first PCR 
positivity. In the absence of positive PCR, the duration will be set to a maximum 
of 28 days;
-
Descriptive statistics (N, geometric mean, SD, 95% CI, range) of parasitaemia at 
first PCR positivity;
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of time to 
SDUDVLWDHPLDRISDUDVLWHVSHUP/EORRG&RKRUWs 1 and 2);
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of parasitaemia at the 
time parasitaHPLDSDUDVLWHVSHUP/EORRG&RKRUWs 1 and 2);
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of time to 
SDUDVLWDHPLDRISDUDVLWHVSHUP/EORRG&RKRUW
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of parasitaemia at the 
time parasitaHPLDSDUDVLWHVSHUP/EORRG&RKRUW
-
Descriptive statistics per cohort (geometric mean, SD, 95% CI, range) of time to 
first dose of treatment with Riamet® (Cohorts 1 and 2); and
-
Descriptive statistics per cohort (geometric mean, SD, 95% CI, range) of 
parasitaemia at first dose of treatment with Riamet® (Cohorts 1 and 2).
The number and proportion of participants with presence of positive PCR and 
SDUDVLWDHPLDRIRUSDUDVLWHVSHUP/EORRGEHWZHHQLQRFXODWLRQZLWK
PfSPZ-DVI Challenge and Day 28 will be summarised per cohort. Corresponding 
two-sided 90% Exact Clopper-Pearson confidence limits will be presented as well.
Secondary endpoints:
To characterise the blood-stage parasite profile, a model will be fitted to the measured 
parasitaemia data prior to Riamet® administration to be able to predict the growth of 
parasites as a function of time. This model may be a log-linear model or, if the 
parasitaemia profile shows a cyclic behaviour due to synchronicity of the parasites and 
their sequestration during the late stages of their lifecycle, it may include a sinus 
function. The latter will allow the estimation of the period of the observed waves in 

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addition to the parasite growth. The following parameters will be calculated from the 
individual fits of the data:
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) 
of parasite growth rate expressed as the PMR48;
-
If cycles are observed and if their estimated period is not 48 h, descriptive 
statistics per cohort and overall (geometric mean, SD, 95% CI, range) of parasite 
growth rate expressed as the PMRLC; and
-
Descriptive statistics (geometric mean, SD, 95% CI, range) of predicted time to 
UHDFKSDUDVLWDHPLDWKUHVKROGRIILUVWSRVLWLYH3&5SDUDVLWHVSHUP/EORRG
SDUDVLWHVSHUP/EORRGIRUCohorts 1 and 2.
To characterise the blood-stage parasite clearance profile of Riamet® in malaria infection 
in naïve healthy participants after PfSPZ-DVI Challenge; by calculating:
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) 
of time to parasite clearance, i.e., time between drug administration and first PCR 
below the limit of quantification;
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) 
of log10PRR48;
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI, range) 
of log10PRR48,max; and
-
Descriptive statistics per cohort and overall (geometric mean, SD, 95% CI range) 
of PC½.
The parasite reduction ratios (PRRs) and the clearance half-life will be estimated from a 
log-linear model that will be fitted to the parasitaemia data observed from the 
administration of the treatment onwards. The PRR48, PRR48,max and PC½ will be 
calculated from the individual parameters and summarised by cohort.
Exploratory endpoints:
To characterise the individual variability of blood-stage parasite profile: 
-
Obtain the inter-individual variability of the parasite growth rate; and
-
5th, 50th and 95th percentiles of predicted time to reach parasitaemia threshold of 
ILUVWSRVLWLYH3&5SDUDVLWHVSHUP/EORRGSDUDVLWHVSHUP/EORRG
including their 95% CI.
Both these estimates will be based on the mixed model on parasite growth data described 
above.
9.1.3
Safety Data
Safety parameters will be tabulated and analysed descriptively.
Adverse Events
The original terms entered in the eSource system by Investigators to identify AEs will be 
fully described and coded according to the Medical Dictionary for Regulatory Activities 
(MedDRA).

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The reported AEs will be allocated to phases based on their start date. All AEs will be 
listed. All AEs with onset after PfSPZ-DVI Challenge will be summarised by cohort.
Summaries will be made per MedDRA primary system organ class, MedDRA preferred 
term, severity, with the number and percentage of participants and the number of events. 
Similar summaries will be prepared for AEs considered to be related to Riamet® and AEs 
considered to be related to the PfSPZ-DVI Challenge, for serious AEs and AEs of special 
interest.
Special attention will be paid to those participants who died, discontinued the 
investigational products due to an AE or experienced a severe or serious AE. Summaries, 
listings and narratives (also see Section 12.11) may be provided, as appropriate.
Malaria Clinical Score 
For the malaria clinical score that will be administered by the PI or his/her trained 
delegate, actual values and changes from baseline will be evaluated by means of 
descriptive statistics. Additionally, expected signs and symptoms (see Attachment 1) will 
be summarised by score.
Clinical Laboratory Tests
Each continuous biochemistry and haematology laboratory test will be evaluated by 
means of descriptive statistics on the actual values, at each assessment time point and by 
cohort. Changes from baseline will also be summarised using descriptive statistics by 
assessment time point and by cohort.
Relative changes in clinical laboratory test values compared to values at baseline will be 
evaluated in accordance with the normal ranges of the clinical laboratory (below, within 
or above normal range). The percentage of participants with clinical laboratory test 
abnormalities will be summarised by cohort.
The number and percentage of participants with liver enzyme elevations after inoculation 
with the PfSPZ-DVI Challenge as defined below will be summarised:
-
ALT or AST >3 x Upper Limit of Normal (ULN);
-
ALT or AST >5 x ULN;
-
ALT or AST >8 x ULN;
-
ALT or AST >3 x ULN and bilirubin >2 x ULN at the same time point, together 
with a conjugated bilirubin fraction> 35% (Potential Hy’s law cases).
A listing of participants with any clinical laboratory test result outside the reference 
ranges will be provided.
Vital Signs
Vital signs parameters will be assessed after at least 10 min in supine position at the time 
points indicated in the Time and Events Schedule. Pulse rate, SBP and DBP will be 
evaluated by means of descriptive statistics (actual values and changes from baseline). 
The percentage of participants with vital signs abnormalities will be summarised by 
cohort in a cross-tabulation of post-baseline versus baseline abnormalities to the normal 
ranges (as defined in Attachment 3).

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Electrocardiogram
Twelve-lead ECG recordings will be performed in triplicate after participants remained 
in a supine position for at least 10 min.
All ECG data automatically measured by ECG devices (PR, QRS, QT, QTcB, QTcF and 
HR) and overall ECG evaluation will be listed. The ECG data, along with changes from 
baseline will be summarised by means of descriptive statistics at each assessment time 
point and by cohort.
The percentage of participants with ECG abnormalities will be summarised by cohort in
a cross-tabulation of post-baseline versus baseline abnormalities to the normal ranges (as 
defined in Attachment 3). This cross-tabulation will include categorical assessment on 
actual values and changes from baseline of QTcB and QTcF prolongation.
Physical Examination
Abnormal findings in physical examination will be listed.
9.2
DETERMINATION OF SAMPLE SIZE
Up to 16 participants will be enrolled in 2 cohorts of 8 participants per cohort. In 
agreement with the Sponsor, additional participants may be recruited in each cohort, to 
replace discontinuations for non-safety reasons and achieve the required cohort size.
This is an exploratory study focusing on the methodology of malaria inoculation in 
healthy participants, thus no formal sample size calculation is performed.

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10.
ADVERSE EVENT REPORTING
10.1
DEFINITIONS
Adverse Event
An AE is any untoward medical occurrence in a clinical study participant administered a 
medicinal (investigational or non-investigational) product. An AE does not necessarily 
have a causal relationship with the treatment. An AE can therefore be any unfavourable 
and unintended sign (including an abnormal finding), symptom or disease temporally 
associated with the use of a medicinal (investigational or non-investigational) product, 
whether or not related to that medicinal (investigational or non-investigational) product.
This includes any occurrence that is new in onset or aggravated in severity or frequency 
from the baseline condition, or abnormal result of diagnostic procedures, including 
clinical laboratory test abnormalities.
Serious Adverse Event
An SAE is any untoward medical occurrence that at any dose meets any of the following 
conditions:
-
results in death;
-
is life-threatening, i.e., the participant was at risk of death at the time of the event
(e.g., ventricular fibrillation and anaphylaxis). The term does not refer to an event 
which hypothetically might have caused death if it were more severe.
-
requires inpatient hospitalization or prolongation of existing inpatient 
hospitalisation:
Hospitalisation refers to an overnight admission into hospital for the purpose of 
investigating and/or treating the AE. Hospitalisation for an elective procedure, or 
routinely scheduled treatment for a pre-existing condition that is unrelated to the 
study and has not worsened, is not an SAE. Treatment on an emergency 
outpatient basis for an event not fulfilling any of the definitions of an SAE given 
above and not resulting in hospital admission, is not an SAE. Cosmetic surgery or 
for social reasons or respite care in the absence of any deterioration in the 
participant’s general condition, is not an SAE; 
-
results in persistent or significant disability/incapacity, i.e., causing substantial 
disruption of the participant’s ability to conduct normal life;
-
is a congenital anomaly/birth defect;
-
is medically significant, i.e., may not be immediately life-threatening or result in 
death or hospitalisation but may jeopardise the participant’s health or may require 
intervention to prevent one of the above outcomes. Examples of such events are 
intensive treatment in an emergency room or at home for allergic bronchospasm,
blood dyscrasias, or convulsions that do not result in hospitalisation or 
development of drug dependency or drug abuse.
-
Constitutes a possible Hy’s Law case (defined as a participant with any value of 
ALT or AST greater than or equal to 3x the Upper Limit of Normal (ULN)

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together with an increase in bilirubin to a value greater than 2xULN [>35% 
direct] and NOT associated with an ALP value greater than 2xULN).
Adverse Events of Special Interest (AESIs)
An AESI (serious or non-serious) is of scientific and medical concern specific to the 
Sponsor’s product or programme, for which ongoing monitoring and rapid 
communication by the Investigator to the Sponsor could be appropriate. Such an event 
might require further investigation in order to characterise and understand it. Depending 
on the nature of the event, rapid communication by the study Sponsor to other parties 
(e.g., regulators) might also be warranted (CIOMS VI, ICH E2F, 2010). For the purpose 
of this study, any abnormalities listed below should be reported as AESI:
Hepatic
-
Any ALT or AST above 3x ULN;
-
Any elevation in bilirubin 2x ULN;
-
Any AST or ALT above 2x ULN AND
o
Total bilirubin level (TBL) >1.5x ULN OR
o
INR >1.4
-
Any AST or ALT above 2x ULN with the appearance of fatigue, nausea, 
vomiting, right upper quadrant pain or tenderness, fever, rash and/or eosinophilia 
(eosinophil count above the ULN).
Cardiac
-
Chest pain
-
High sensitive troponin T 12 ng/L;
-
QTcB or QTcF at any time >480 ms;
-
Bundle branch block (except right bundle branch block that was present prior to 
Riamet® administration);
-
Any arrhythmia, except:
o
sinus bradycardia that is clinically asymptomatic, and not associated with 
any other relevant ECG abnormalities;
o
sinus tachycardia that is clinically asymptomatic, and associated with a 
body temperature >38.0 °C, and not associated with any other relevant 
ECG abnormalities;
o
respiratory sinus arrhythmia;
o
wandering atrial pacemaker; 
o
isolated, single premature atrial/ventricular complex (i.e., no bigeminy, 
trigeminy, couplets, triplets or salvos) that does not occur more than once 
in a particular ECG tracing. 

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Haematological
-
Haemoglobin drop >2 g/dL and under lower limit of normal from baseline prior 
to inoculation; 
-
Absolute neutrophil count <1000/μL;
-
Platelet count <100000/mm3.
Dermatological:*
-
Any suspected cutaneous AE, e.g., rash
* if one of these cutaneous reactions is observed and when feasible, pictures of the 
lesions should be obtained.
Unlisted (Unexpected) Adverse Event
An AE is considered unlisted if the nature or severity is not consistent with the applicable 
product reference safety information (Investigator’s Brochure for an unapproved IMP or 
SmPC or approved Package Insert for an authorized product).
For this protocol, the reference documents for the assessment of expectedness for 
PfSPZ-DVI Challenge is the Investigator’s Brochure [9]. Expected reactions to the 
antimalarial therapies are described in the product’s prescribing information [22,12]:
-
PfSPZ-DVI Challenge can be associated with itching, pain and swelling at the site 
of inoculation. This usually resolves within 24-48 hours.
-
Riamet® (artemether and lumefantrine) may be associated with the following 
AEs; these may occur with a varying periodicity:

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Frequency of Undesirable Effects of Riamet®
Adults and adolescents above 12 years of age
Immune system disorders
Hypersensitivity
Not known
Metabolism and nutrition disorders
Decreased appetite
Very common
Psychiatric disorders
Sleep disorders
Very common
Insomnia
Common
Nervous system disorders
Headache
Very common
Dizziness
Very common
Paraesthesia
Common
Ataxia, hypoaesthesia
Uncommon
Somnolence
Uncommon
Clonus
Common
Cardiac disorders
Palpitations
Very common
Electrocardiogram QT prolonged
Common
Respiratory, thoracic and mediastinal disorders
Cough
Common
Gastrointestinal disorders
Vomiting
Very common
Abdominal pain
Very common
Nausea
Very common
Diarrhoea
Common
Hepatobiliary disorders
Liver function tests increased
Uncommon
Skin and subcutaneous tissue disorders
Rash
Common
Pruritus
Common
Urticaria
Uncommon
Angioedema*
Not known
Musculoskeletal and connective tissue disorders
Arthralgia
Very common
Myalgia
Very common
General disorders and administration site conditions
Asthenia
Very common
Fatigue
Very common
Gait disturbance
Common

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-
Malarone® (antimalarial therapy if an intolerance or contraindication to Riamet®
develops: atovaquone and proguanil hydrochloride) may be associated with the 
following AEs; these may occur with a varying periodicity:
System Organ 
Class
Very 
Common
Common
Uncommon
Rare
Not known2
Blood and 
lymphatic 
disorders 
Anaemia 
Neutropenia 1
Pancytopenia 
Immune system 
disorders 
Allergic reactions 
Angioedema3
Anaphylaxis (see 
section 4.4)
Vasculitis3
Metabolism and 
nutrition disorders 
Hyponatraemia1
Anorexia 
Elevated 
amylase levels1
Psychiatric 
disorders 
Abnormal dreams 
Depression 
Anxiety 
Hallucinations 
Panic attack 
Crying 
Nightmares 
Psychotic disorder 
Nervous system 
disorders 
Headache 
Insomnia
Dizziness 
Seizure 
Cardiac disorders 
Palpitations 
Tachycardia 
Gastrointestinal 
disorders 
Nausea1
Vomiting 
Diarrhoea 
Abdominal 
pain 
Stomatitis 
Gastric 
intolerance3
Oral ulceration3
Hepatobiliary 
disorders 
Elevated liver 
enzymes1
Hepatitis 
Cholestasis3
Skin and 
subcutaneous 
tissue disorders 
Pruritus 
Rash 
Hair loss 
Urticaria 
Stevens-Johnson 
Syndrome 
Erythema 
multiforme 
Blister 
Skin exfoliation 
Photosensitivity 
reactions 
General disorders 
and administration 
site conditions 
Fever 
Respiratory, 
thoracic and 
mediastinal 
disorders 
Cough 

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An Inoculum-Related AE is a sign or symptom associated with malaria infection, i.e., 
confirmed by a protocol-defined PCR positivity (defined for the purpose of this study as 
DT3&5RXWFRPH parasites per mL blood) at
-
the onset of the event; or
-
if the event started within 1 day of parasite clearance (defined as a qPCR value of 
0 parasites per mL blood after initiating antimalarial therapy) or 
laboratory-defined PCR positivity (a qPCR outcome of 50-249 parasites per mL 
blood).
Close monitoring of expected signs and symptoms associated with malaria infection 
(listed in Attachment 1) form part of the safety evaluation of the study and will be used 
as part of the decision criteria to administer registered antimalarial therapy. These events 
will be classified as AEs, and may also be further sub-classified as inoculum-related AEs 
(see Section 5.1, Section 7.3.1, Section 7.3.2 and Section 9.1.3).
An Antimalarial Therapy-Related AE is a sign or symptom associated with the 
antimalarial drugs. Expected reactions to the antimalarial drugs are described in the 
product’s prescribing information [22,12].
10.2
INTENSITY OF ADVERSE EVENTS
Each AE must be rated on a 5-point scale of increasing intensity according to the 
Common Terminology Criteria for Adverse Events (CTCAE) version 5.0:
Note: the semi-colon within the description of the grade indicates ‘or’.
Grade 1:
Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; 
intervention not indicated.
Grade 2:
Moderate; minimal, local or non-invasive intervention indicated; limiting age-appropriate
instrumental activities of daily life (preparing meals, shopping for groceries or clothes, 
using the telephone, managing money, etc.).
Grade 3:
Severe or medically significant but not immediately life-threatening; hospitalisation or 
prolongation of hospitalisation indicated, disabling; limiting self-care activities of daily 
life (bathing, dressing and undressing, feeding self, using the toilet, taking medications 
and not bedridden).
Grade 4:
Life-threatening consequences; urgent intervention indicated.
Grade 5:
Death related to AE.
10.3
CAUSALITY ASSESSMENT
The Investigator must assess the relationship of each event to the PfSPZ-DVI Challenge
agent and the antimalarial treatments (separately) and decide whether, in his or her 

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medical judgement, there is a reasonable possibility that the event may have been caused 
by any of the study agents. Where possible, a distinction should be made between events 
considered related to the PfSPZ-DVI Challenge agent and the antimalarial treatments. If 
there is no valid reason for suggesting a relationship, then the AE should be classified as 
“not related”. Alternatively, if there is any valid reason for suspecting a possible 
cause-and-effect relationship between the investigational product(s) and the occurrence 
of the AE (even if undetermined or untested), then the AE should be considered as 
“related” to whichever product as relevant. This should be documented in the 
participant’s source document and eSource system.
The following may guide this assessment:
-
Related – the temporal relationship between the event and the administration of 
the PfSPZ-DVI Challenge agent and/or antimalarial treatment is compelling 
and/or follows a known or suspected response pattern to that product, and the 
event cannot be explained by the participant’s medical condition, other therapies 
or accident;
-
Not related – the event can be readily explained by other factors such as the 
participant’s underlying medical condition, concomitant therapy or accident and 
no plausible temporal or biologic relationship exists between the PfSPZ-DVI 
Challenge agent and/or antimalarial treatment and the event.
In addition to the assessments of relationship to the investigational products, the 
Investigator should comment on the adverse event record in the eSource system whether 
an AE is related to the study participation of the participant (study procedures etc.).
10.4
ACTION TAKEN REGARDING THE INVESTIGATIONAL PRODUCTS
The action taken towards the investigational products must be described as follows:
-
Permanently discontinued;
-
No action taken;
-
Unknown/Not applicable.
10.5
OUTCOME
The outcome of each AE must be rated as follows:
-
Recovered/resolved;
-
Recovering/resolving;
-
Not recovered/not resolved;
-
Recovered with sequelae/resolved with sequelae;
-
Fatal;
-
Unknown.

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10.6
RECORDING OF ADVERSE EVENTS
All (S)AEs occurring during the clinical investigation (i.e., monitored continuously from 
informed consent until the last study-related activity at the EOS on Day 28) must be 
documented in the eSource System.
Whenever possible, diagnoses should be given when signs and symptoms are due to a 
common etiology (e.g., cough, runny nose, sneezing, sore throat, and head congestion 
should be reported as “upper respiratory infection”). Investigators must record their 
opinion concerning the relationship of the (S)AE to the investigational products in the 
eSource System. All measures required for (S)AE management must be recorded in the 
source documents and reported according to Sponsor’s instructions.
All AEs occurring at any time during the study (including the follow-up period) will be 
followed by the Investigator until satisfactory resolution (e.g., value back to baseline 
value) or stabilisation or until final database lock. If necessary, in order to obtain 
additional information to ensure safety to the participant, additional blood and urine 
samples may be taken at the discretion of the Investigator. Certain long-term AEs related 
to therapy cannot be followed until resolution within the setting of this study. In these 
cases, follow-up will be the responsibility of the treating physician.
10.7
REPORTING OF SERIOUS ADVERSE EVENTS TO PRIMEVIGILANCE 
LTD.
All SAEs and AESIs, independent of the circumstances or suspected cause, must be 
reported on a Serious Adverse Event Form by the investigator to PrimeVigilance Ltd.
within 24 h of their knowledge of the event, preferably by fax (+44 800 471 5694) or by 
e-mail (MMV@primevigilance.com).
The SAE form should include a clearly written narrative describing signs, symptoms, and 
treatment of the event, diagnostic procedures, as well as any relevant laboratory data and 
any sequelae, in order to allow a complete medical assessment of the case and 
independent determination of the possible causality.
Follow-up and outcomes should be reported for all participants who experience an SAE.
It is critical that the information provided on the Serious Adverse Event Form matches 
the information recorded in the source documents and in the eSource system for the same 
event.
Copies of additional laboratory tests, consultation reports, postmortem reports, hospital 
case reports, autopsy reports, and other documents should be sent when requested and 
applicable. Follow-up reports relative to the participant’s subsequent course must be 
submitted to PrimeVigilance Ltd. until the event has subsided or, in case of permanent 
impairment, until the condition stabilises.
10.8
PREGNANCY
All initial reports of pregnancy in participants or in partners of male participants must be 
reported by the Investigator to PrimeVigilance Ltd. within 24 h of his/her knowledge of 
the event using a Pregnancy Form. Any participant who becomes pregnant during the 
study must be promptly withdrawn from the study (cfr. Section 8).

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The Investigator will contact the participant at the expected time of delivery for follow-
up. Abnormal pregnancy outcomes (e.g., spontaneous abortion, stillbirth, neonatal death, 
congenital abnormality, birth defect) are considered SAEs and must be reported using the 
Serious Adverse Event Form.
10.9
REPORTING OF SERIOUS ADVERSE EVENTS TO COMPETENT 
AUTHORITIES/ETHICS COMMITTEES
PrimeVigilance Ltd. assumes responsibility for appropriate reporting of AEs to the 
regulatory authorities. PrimeVigilance Ltd. will also report to the Investigator all SAEs 
that are unlisted (unexpected) and associated with the use of the drug. The Investigator
(or PrimeVigilance Ltd. where required) must report these events to the appropriate 
Independent Ethics Committee/Institutional Review Board (IEC/IRB) that approved the 
protocol, unless otherwise required and documented by the IEC/IRB.
Adverse events reporting, including suspected unexpected serious adverse reactions
(SUSARs), will be carried out in accordance with applicable local regulations.
After termination of the clinical study (determined as LPLV), any unexpected safety 
issue that changes the risk-benefit analysis and is likely to have an impact on the 
participants who have participated in the study, together with proposed actions, will be 
reported by the Sponsor/PrimeVigilance Ltd. to the competent authority(ies) concerned
as soon as possible.

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11.
ETHICAL ASPECTS
11.1
STUDY-SPECIFIC DESIGN CONSIDERATIONS
Potential participants will be fully informed of the nature of the study and of the risks and 
requirements of the study before any study-related assessment will be carried out. During 
the study, participants will be given any new information that may affect their decision to 
continue participation. They will be informed that their participation in the study is 
voluntary and that they may withdraw from the study at any time with no reason given 
and without penalty or loss of benefits to which they would otherwise be entitled. Only 
participants who are fully able to understand the risks, benefits, and potential AEs of the 
study, and who provide their consent voluntarily will be enrolled in the study.
11.2
REGULATORY ETHICS COMPLIANCE
11.2.1
Investigator Responsibilities
The Investigator(s) should be qualified by education, training, and experience to assume 
responsibility for the proper conduct of the study, should meet all the qualifications 
specified by the applicable regulatory requirement(s), and should provide evidence of 
such qualifications through up-to-date curriculum vitae or other relevant documentation 
requested by the Sponsor, the IRB/IEC or the regulatory authority(ies).
The Investigator is responsible for ensuring that the clinical study is performed in 
accordance with the protocol, current International Council for Harmonization of 
Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines on Good 
Clinical Practice (GCP), and applicable regulatory and country-specific requirements.
Good Clinical Practice is an international ethical and scientific quality standard for 
designing, conducting, recording, and reporting studies that involve the participation of 
human participants. Compliance with this standard provides public assurance that the 
rights, safety, and well-being of study participants are protected, consistent with the 
principles originating from the Declaration of Helsinki (1964 and revisions), and that the 
clinical study data are credible.
11.2.2
Independent Ethics Committee or Institutional Review Board 
(IEC/IRB)
An IRB/IEC should safeguard the rights, safety, and well-being of all study participants. 
Special attention should be paid to studies that may include vulnerable participants.
Before the start of the study, the Investigator (or Sponsor where required) will provide 
the IEC/IRB with current and complete copies of the following documents:
-
final protocol and, if applicable, amendments;
-
Sponsor-approved ICF (and any updates or any other written materials to be 
provided to the participants);
-
Investigator’s Brochure (or equivalent information) and addenda;
-
available safety information;

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-
information on compensation for study-related injuries or payment to participants
for participation in the study, if applicable;
-
Investigator’s current curriculum vitae or other documentation evidencing 
qualifications (unless not required, as documented by the IEC/IRB);
-
information regarding funding, name of the Sponsor, institutional affiliations, 
other potential conflicts of interest and incentives for participants;
-
any other documents that the IEC/IRB may require to fulfill its obligation.
This study will be undertaken only after the IEC/IRB has given full written approval of 
the final protocol and amendments (if any), the ICF(s) and updates (if any), applicable 
recruiting materials and any other written information to be provided to the participants,
and the Sponsor has received a copy of this approval. This approval letter must be dated 
and must clearly identify the IEC/IRB and the documents being approved.
During the study, the Investigator (or Sponsor where required) will send the following 
documents and updates to the IEC/IRB for its review and approval, where appropriate: 
-
protocol amendments;
-
revision(s) to the ICF and any other written materials to be provided to the 
participants;
-
revisions to compensation for study-related injuries or payment to participants for 
participation in the study;
-
Investigator’s Brochure addenda or new edition(s);
-
summaries of the status of the study at intervals stipulated in guidelines of the 
IEC/IRB (at least annually);
-
reports of AEs that are serious, unlisted, and associated with the IMP;
-
new information that may adversely affect the safety of the participants or the 
conduct of the study;
-
deviations from or changes to the protocol to eliminate immediate hazards to the 
participants;
-
report of death of any participants under the Investigator’s care;
-
notification if a new Investigator is responsible for the study at the clinical site;
-
Development Safety Update Report, Short-Term Study Specific Safety Summary 
and Line Listings, where applicable;
-
any other requirements of the IEC/IRB.
For all protocol amendments (excluding the ones that are purely administrative, with no 
consequences for participants, data or study conduct), the amendment and applicable ICF
revisions must be submitted promptly to the IEC/IRB for review and approval before 
implementation of the change(s), except when necessary to eliminate immediate hazard 
to the study participants. If a deviation from or a change to the protocol was implemented 
to eliminate an immediate hazard to study participants, then the implemented deviation 
or change, the reasons for it, and, if appropriate, the protocol amendment should be 
submitted to the IEC/IRB as soon as possible.

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The Investigator (or Sponsor where required) will notify the IEC/IRB about the study 
completion within 90 days after the end of the study (defined as LPLV).
11.2.3
Informed Consent
Each participant must give written consent according to local requirements after the 
nature of the study has been fully explained. The consent form must be signed before 
performance of any study-related activity. The consent form that is used must be 
approved by both the Sponsor and the reviewing IEC/IRB. The informed consent should 
be in accordance with the principles that originated in the Declaration of Helsinki, 
current ICH and GCP guidelines, applicable regulatory requirements, and Sponsor
policy.
Before enrolment in the study, the Investigator or an authorised member of the clinical 
staff must explain to potential participants the aims, methods, reasonably anticipated 
benefits and potential hazards of the study, and any discomfort participation in the study 
may entail. Participants will be informed that their participation is voluntary and that they 
may refuse to participate or withdraw consent to participate at any time, without penalty 
or loss of benefits to which the participant was entitled. Finally, they will be told that the 
Investigator will maintain a participant identification register for the purposes of long-
term follow-up if needed and that their records may be accessed by health authorities and 
authorised Sponsor staff without violating the confidentiality of the participant, to the 
extent permitted by the applicable law(s) or regulations. By signing the ICF the 
participant is authorising such access, and agrees to allow his or her study physician to 
recontact the participant for the purpose of obtaining consent for additional safety 
evaluations, if needed.
The language about the study used in the oral and written information, including the ICF, 
should be non-technical and practical and should be understandable to the participant (or 
the participant’s legally acceptable representative). The participant will be given 
sufficient time to read the ICF and the opportunity to ask questions. After this 
explanation and before entry into the study, consent should be appropriately recorded by 
means of the participant's personally dated signature. After having obtained consent, a 
copy of the ICF must be given to the participant.
If a participant (or legally acceptable representative) is unable to read or write, an 
impartial witness should be present during the entire informed consent discussion. After 
the written ICF and any other written information to be provided to the participants, is 
read and explained to the participant (or legally acceptable representative), and after the 
participant (or legally acceptable representative) has orally consented to the participant’s 
participation in the study and, if capable of doing so, has personally dated and signed the 
ICF, the witness should personally date and sign the consent form. By signing the ICF, 
the witness attests that the information in the ICF and any other written information was 
accurately explained to, and apparently understood by, the participant (or legally 
acceptable representative), and that informed consent was freely given by the participant
(or legally acceptable representative).

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11.2.4
Privacy of Personal Data
The collection and processing of personal data from participants enrolled in the study 
will be limited to those data that are necessary to investigate the safety, quality and utility 
of the IMP used in the study.
These data must be collected and processed with adequate precautions to ensure 
confidentiality and compliance with applicable data privacy protection laws and 
regulations. Appropriate technical and organizational measures to protect the personal 
data against unauthorized disclosures or access, accidental or unlawful destruction, or 
accidental loss or alteration must be put in place. Sponsor personnel whose 
responsibilities require access to personal data need to agree to keep the identity of the 
study participants confidential.
During the informed consent process the participants will be informed that, if they 
participate in this study, the Investigator may be required to allow direct access to 
participants’ original medical records for study-related monitoring, audit, IEC/IRB 
review, and regulatory inspection. The participants will also be informed of possible 
transfer of the collected personal data to other entities and to other countries and that 
should such transfer take place, the Sponsor will ensure that technical and organisational 
measures are put in place to protect the data.

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12.
ADMINISTRATIVE REQUIREMENTS
12.1
PROTOCOL AMENDMENTS
Neither the Investigator nor the Sponsor will modify this protocol without a formal 
amendment. All protocol amendments must be issued by the Sponsor and signed and 
dated by the Investigator. Protocol amendments must not be implemented without prior 
IEC/IRB approval nor when the relevant competent authority has raised any grounds for 
non-acceptance, except when necessary to eliminate immediate hazard to the 
participants, in which case an amendment must be promptly submitted to the IEC/IRB 
and relevant competent authority. Documentation of amendment approval by the 
Investigator and IEC/IRB must be provided to the Sponsor or his designee. When the 
change(s) involves only logistic or administrative aspects of the study, the IRB (and IEC 
where required) only needs to be notified.
12.2
PARTICIPANT IDENTIFICATION, ENROLMENT AND SCREENING 
LOGS
The Investigator agrees to complete a participant identification and enrolment log to 
permit easy identification of each participant during and after the study. This document 
will be reviewed by the Sponsor site contact for completeness.
The participant identification and enrolment log will be treated as confidential and will 
be filed by the Investigator in the study file. To ensure participant confidentiality, no 
copies will be made. All reports and communications related to the study will identify 
participants by initials and/or assigned number only.
The Investigator must also complete a participant screening log which reports on all 
participants who were seen to determine eligibility for inclusion in the study.
12.3
SOURCE DOCUMENTATION
The Clinspark system is an electronic data capturing and information management 
system that will also serve as an eSource system for this study. The system combines all 
aspects of source data capturing with process control and clinical study management. All 
clinical and laboratory data, except those that are paper-based, will be collected directly 
in Clinspark. The Source Document Identification Overview will specify which 
information will be eSource and which will be paper-based. The monitor will check data 
at the monitoring visits to the clinical site. The Investigator will ensure that the data 
collected are accurate, complete and legible. Data will be monitored within Clinspark by
the study monitor who has only reading rights. Any changes required following 
monitoring will be made by site personnel or the Investigator and will be documented 
with a full audit trail within Clinspark.
At a minimum, source documentation must be available for the following: participant
identification, eligibility and study identification; date of informed consent, dates of 
visits, results of safety and efficacy parameters as required by the protocol, record of all 
AEs, follow-up of AEs, concomitant medication, drug receipt/dispensing/return records, 
investigational product administration information, laboratory and ECG printouts (if not 
available digitally), date of study completion and reason for early discontinuation of 
investigational products or withdrawal from the study, if applicable.

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It is recommended that the author of an entry in the (e)Source documents be identifiable.
Source data may be directly captured from devices, transferred from third parties (e.g., 
laboratory data), or entered manually into the eSource system in use at the clinical site. In 
such case, the majority of the source data will only be available electronically. The 
remainder of the data, captured initially on paper, may be entered retrospectively into the 
eSource system.
Following the ICH-GCP guidelines, direct access to (e)Source documentation (medical 
records) must be allowed.
12.4
CASE REPORT FORM COMPLETION
All source data, except those that are paper-based, will be collected directly into the 
eSource system. Data must be entered in English. Paper-based source data will be 
manually transcribed to the eSource system. Only the data required for the clinical 
database will be transferred electronically from the eSource system to the clinical 
database. The Investigator must verify that all data entries in the eSource system are 
accurate and correct.
12.5
MONITORING
Medical and clinical monitoring of the study will be done under the responsibility of the 
Sponsor by ICON Clinical Research and IQVIA RDS & Integrated Services Belgium 
NV/SA, respectively.
The monitor will perform on-site monitoring visits as frequently as necessary. The 
monitor will record the dates of the visits in a study site visit log that will be kept at the 
clinical site. The first post initiation visit will be made as soon as possible after enrolment 
has begun. At these visits, the monitor will compare the data captured in the eSource
system for completeness and accuracy and perform data source verification to any data 
that has been captured as paper source or entered in the system later on. The nature and 
location of all source documents will be identified to ensure that all sources of original 
data required to complete the eSource system are known to the Sponsor and clinical staff 
and are accessible for verification by the Sponsor site contact. If electronic records are 
maintained at the investigational site, the method of verification must be discussed with 
the clinical staff.
Direct access to eSource documentation (medical records) must be allowed at all times 
for the purpose of verifying that the data recorded in the eSource system are consistent 
with the source documentation data. Findings from this review of captured data will be 
discussed with the clinical staff. During on-site monitoring visits (notified and agreed in 
advance with the clinical staff), the relevant clinical staff will be available, the (e)Source 
documentation will be accessible, and a suitable environment for review of study-related 
documents will be provided. The monitor will meet with the Investigator on a regular 
basis during the study to provide feedback on the study conduct.
12.6
DATA MANAGEMENT
Data management of the study will be performed under Sponsor delegation by SGS Life 
Sciences.

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After the data entered in the eSource system are released by the Investigator, the data will 
be uploaded into the clinical database to perform cleaning activities. Computerised data 
cleaning rules will be used in addition to manual review to check for discrepancies and to 
ensure consistency and completeness of the data. Queries emerging during data cleaning 
will be generated by the clinical data manager in the eSource system. The Investigator or 
his designee will answer the queries and update the source data, if needed.
The clinical database will be locked as soon as it is considered clean. Before the clinical 
database will be locked, the study eSource system will be locked by the clinical staff.
Only authorised and well-documented updates to the study data are possible after 
database lock. The locked database is used in the final statistical analysis for study 
reporting. Measures will be undertaken to protect participant data handed over by the 
Investigator to the data management department and during inspections against 
disclosure to unauthorised third parties. Participant confidentiality will be maintained at 
all times.
12.7
DATA QUALITY ASSURANCE
The accuracy and reliability of the study data will be assured by the selection of qualified 
Investigators and appropriate study sites, review of protocol procedures with the 
Investigator and associated personnel prior to the study, and by periodic monitoring visits 
by the Sponsor or designate.
The Sponsor or his designee will review the eSource system for accuracy and 
completeness during (on-site) monitoring visits and after transmission to the Sponsor;
any discrepancies will be resolved with the Investigator or designee, as appropriate. After 
upload of the data into the clinical study database, their accuracy is verified using 
appropriate validation programs.
In accordance with Good Clinical Research Practice Guidelines and Recommendations, 
the Sponsor will be entitled to audit the facilities used in the clinical and laboratory parts 
of the study, as well as to access all the data files pertaining to the study. Similar 
procedures may also be conducted by agents of any regulatory body, either as part of a 
national GCP compliance program or to review the results of this study in support of a 
regulatory submission. The Investigator should immediately notify the Sponsor if they 
have been contacted by a regulatory agency concerning an upcoming inspection.
12.8
ON-SITE AUDITS
Representatives of the Sponsor’s quality assurance department may visit the clinical site 
at any time during or after completion of the study to conduct an audit of the study in 
compliance with regulatory guidelines and company policy. These audits will require 
access to all study records, including source documents, for inspection and comparison 
with the eSource system. Participant privacy must, however, be respected. The 
Investigator and clinical staff are to be present and available for consultation during 
routinely scheduled site audit visits conducted by the Sponsor or his designee.
Similar procedures may also be conducted by agents of any regulatory body, either as 
part of a national GCP compliance program or to review the results of this study in 
support of a regulatory submission. The Investigator should immediately notify the 

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Sponsor if they have been contacted by a regulatory agency concerning an upcoming 
inspection.
12.9
STUDY TERMINATION
The Sponsor has the right to terminate the study at any time. In case of an early 
termination of the study for safety reasons, or temporary halt by the Sponsor, the 
IEC/IRB and competent authorities should be notified within 15 calendar days and 
should be provided with a detailed written explanation for the termination/halt.
An end-of-study declaration will be submitted to the regulatory authorities and IEC/IRB
after the complete study has ended. This notification will be submitted within 90 days 
after the end of the study.
12.10 RECORD RETENTION
In compliance with the ICH/GCP guidelines, the Investigator/Institution will maintain all 
eSource and all paper source documents that support the data collected from each 
participant, as well as all study documents as specified in ICH/GCP Section 8, Essential 
Documents for the Conduct of a Clinical Trial, and all study documents as specified by 
the applicable regulatory requirement(s). The Investigator/Institution will take measures 
to prevent accidental or premature destruction of these documents.
Essential documents must be retained until at least 2 years after the last approval of a 
marketing application in an ICH region and until there are no pending or contemplated 
marketing applications in an ICH region or until at least 2 years have elapsed since the 
formal discontinuation of clinical development of the IMP. These documents will be 
retained for a longer period if required according to the applicable regulatory 
requirements or per agreement with the Sponsor. It is the responsibility of the Sponsor to 
inform the Investigator/Institution as to when these documents no longer need to be 
retained.
If the responsible Investigator retires, relocates, or for any other reasons withdraws from 
his responsibility of keeping the study records, custody must be transferred to a person 
who will accept the responsibility. The Sponsor must be notified in writing of the name 
and address of the new custodian. Under no circumstance shall the Investigator relocate 
or dispose of any study documents without having obtained written approval from the 
Sponsor.
If it becomes necessary for the Sponsor or the appropriate regulatory authority to review 
any documentation related to the study, the Investigator must permit access to such 
reports.
12.11 USE OF INFORMATION AND PUBLICATION
All information, including but not limited to, information regarding investigational 
product or the Sponsor’s operations (e.g., patent application, formulas, manufacturing 
processes, basic scientific data, prior clinical data, formulation information) supplied by 
the Sponsor to the Investigator and not previously published, and any data generated as a 
result of this study are considered confidential and remain the sole property of the 
Sponsor. The Investigator agrees to maintain this information in confidence, to use this 

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information only to accomplish this study, and not to use it for other purposes without the 
Sponsor’s prior written consent.
The Investigator understands that the information generated in this clinical study will be 
used by the Sponsor in connection with the continued development of the investigational 
product, and thus may be disclosed as required to other clinical Investigators or 
regulatory agencies. To permit information derived from the clinical studies to be used, 
the Investigator is obliged to provide the Sponsor with all data obtained in the study.
The results of the study will be reported in a Clinical Study Report generated under the 
responsibility of the Sponsor and will contain eSource system data from all clinical sites 
that participated in the study. 
Clinical narratives may be written for the following events (for example):
-
All deaths (irrespective of drug relationship);
-
All other SAEs and AESIs during treatment with the investigational products;
-
All discontinuations of the investigational products due to AEs (irrespective of 
drug relationship);
-
At the discretion of the team and after statistical analysis of the data, certain 
discontinuations not related to AEs or treatment failure, i.e., related to lost to 
follow-up or withdrawal of consent (irrespective of treatment group);
-
Any events of special interest explicitly requested by the regulatory agencies.
The PI will sign off the final version of the Clinical Study Report. A summary of this 
final version will be provided to the Investigators, the applicable regulatory authorities, 
and the IECs/IRBs, if required by the applicable regulatory requirements, within 1 year 
after the end of the study (LPLV).
The Sponsor shall have the right to publish study data and information without approval 
from the Investigator. If an Investigator wishes to publish information from the study, a 
copy of the manuscript must be provided to the Sponsor for review at least 60 days 
before submission for publication or presentation. Expedited reviews will be arranged for 
abstracts, poster presentations or other materials. If requested by the Sponsor in writing, 
the Investigator will withhold such publication for up to an additional 60 days to allow 
for filing of a patent application. In the event that issues arise regarding scientific 
integrity or regulatory compliance, the Sponsor will review these issues with the 
Investigator. The Sponsor will not mandate modifications to scientific content and does 
not have the right to suppress information. For multicentre study designs and substudy 
approaches, results may need to be published in a given sequence (e.g., substudies should 
generally not be published before the primary endpoints of a study have been published). 
Similarly, Investigators will recognise the integrity of a multicentre study by not 
publishing data derived from an individual clinical site until the combined results from 
the completed study have been published in full, within 12 months after conclusion, 
abandonment or termination of the study at all clinical sites, or the Sponsor confirms 
there will be no multicentre study publication. Authorship of publications resulting from 
this study will be based on the guidelines on authorship, such as those described in the 
Uniform Requirements for Manuscripts Submitted to Biomedical Journals, which state 
that the named authors must have made a significant contribution to the design of the 

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study or analysis and interpretation of the data, provided critical review of the paper, and 
given final approval of the final version.
12.12 REGISTRATION OF CLINICAL STUDIES AND DISCLOSURE OF 
RESULTS
Public disclosure of the study is under the responsibility of the Sponsor. The study will 
be registered on the ClinicalTrials gov site.
12.13 CONFIDENTIALITY
All study documents are provided by the Sponsor to the Investigator and appointed 
clinical staff in confidence. None of this material may be disclosed to any party not 
directly involved in the study without the Sponsor’s written permission.
The Investigator must assure that participants’ anonymity will be maintained. The 
Investigator will keep a separate list with at least the initials, the participants’ study 
numbers, names, addresses, and telephone numbers. The Investigator will maintain this 
for the longest period of time allowed by his/her own institution and, in any case, until 
further communication from the Sponsor.

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13.
REFERENCES
1. Bastiaens GJH, van Meer MPA, Scholzen A, et al. Safety, Immunogenicity, and 
Protective Efficacy of Intradermal Immunization with Aseptic, Purified, 
Cryopreserved Plasmodium falciparum Sporozoites in Volunteers Under 
Chloroquine Prophylaxis: A Randomized Controlled Trial. Am J Trop Med Hyg. 
2016 Mar;94(3):663-73.
2. Chughlay MF, El Gaaloul M, Donini C, et al. A randomised, double-blind, 
placebo-controlled, Phase Ib study to evaluate the safety, tolerability and 
chemoprotective antimalarial activity of P218 against controlled human malaria 
infection by direct venous inoculation (DVI) of Plasmodium falciparum 
sporozoite (PfSPZ-DVI) in non-immune healthy adult volunteers. 68th Meeting, 
American Society for Tropical Medicine and Hygiene. November 20-24th, 2019. 
Gaylord National Resort and Convention Center National Harbor, MD, USA.
3. Collins KA, Rückle T, Elliott S, et al. DSM265 400 mg clears asexual stage 
parasites but not mature gametocytes from the blood of healthy participants
experimentally infected with Plasmodium falciparum. Antimicrobial Agents and 
Chemotherapy 2019;63:1-11.
4. Collins KA, Wang C, Adams M, et al. Human to mosquito transmission during 
controlled human malaria infection: a model to evaluate transmission blocking 
interventions. J Clin Invest 2018;128:1551-1562.
5. Fridericia LS. The duration of systole in the electrocardiogram of normal 
participants and of patients with heart disease. Acta Med Scand. 1920;53:469–
486.
6. Gaur A, McCarthy JS, Panetta JC, et al. Safety, tolerability, pharmacokinetics, 
and antimalarial efficacy of a novel Plasmodium falciparum ATP4 inhibitor 
(SJ733): a first-in-human and induced blood stage malaria phase 1a/ 1b study. 
Lancet ID 2019; accepted for publication.
7. Gómez-Pérez GP, Legarda A, Muñoz J, et al. Controlled human malaria infection 
by intramuscular and direct venous inoculation of cryopreserved Plasmodium 
falciparum sporozoites in malaria-naive volunteers: effect of injection volume 
and dose on infectivity rates. Malar J. 2015 Aug 7;14:306. 
8. Hodgson SH, Juma E, Salim A, et al. Evaluating controlled human malaria 
infection in Kenyan adults with varying degrees of prior exposure to Plasmodium 
falciparum using sporozoites administered by intramuscular injection. Front 
Microbiol. 2014 Dec 12;5:686. 
9. Investigational Brochure PfSPZ Challenge (NF54). Sanaria Incorporated, 
Edition 13, 7 March 2019. 
10. Lell B, Mordmüller B, Dejon Agobe JC, et al. Impact of Sickle Cell Trait and 
Naturally Acquired Immunity on Uncomplicated Malaria after Controlled Human 
Malaria Infection in Adults in Gabon. Am J Trop Med Hyg. 2018 
Feb;98(2):508-515.

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11. Lyke K, Laurens M, Strauss K, et al. Optimizing Intradermal Administration of 
Cryopreserved Plasmodium falciparum Sporozoites in Controlled Human Malaria 
Infection. Am J Trop Med Hyg. 2015 Dec;93(6):1274-84.
12. Malarone Summary of Product Characteristics (May 2017). GlaxoSmithKline 
Pharmaceuticals SA/NV. Wavre, Belgium. (bijsluiters.fagg-
afmps.be/registrationSearchServlet?key=BE186821&leafletType=skp). Accessed 
November 2019.
13. McCarthy JS, Donini C, Chalon S, et al. Defining the antimalarial activity of the 
novel Plasmodium phosphatidylinositol 4-kinase inhibitor MMV390048 in 
healthy volunteers. In preparation.
14. McCarthy J, Donini C, Chalon S, et al. Antimalarial activity of the Plasmodium 
phosphatidylinositol 4-kinase inhibitor MMV048 in the human Induced Blood 
Stage Malaria model. 28th European Congress of Clinical Microbiology and 
Infectious Diseases. April 21-24, 2018. Madrid, Spain.
15. McCarthy JS, Sekuloski S, Griffin PM, et al. A Pilot Randomised Trial of 
Induced Blood-Stage Plasmodium falciparum Infections in Healthy Volunteers 
for Testing Efficacy of New Antimalarial Drugs. PlOS One. 2011,6(8):e21914.
16. Mira-Martinez S, van Schuppen E, Amambua-Ngwa A, et al.Expression of the 
Plasmodium falciparum Clonally Variant clag3 Genes in Human Infections. J 
Infect Dis. 2017,215(6):938-945.
17. Mira-Martinez S, Pickford AK, Rovira-Graells N, et al. Identification of 
Antimalarial Compounds That Require CLAG3 for Their Uptake by Plasmodium 
falciparum-Infected Erythrocytes. Antimicrob Agents Chemother. 2019,63(5).
18. Mordmüller B, Supan C, Sim K L, et al. Direct venous inoculation of
Plasmodium falciparum sporozoites for controlled human malaria infection: a 
dose-finding trial in two centres Malar J. 2015 Mar 18; 14:117.
19. Mordmüller B, Surat G, Lagler H, et al. Sterile protection against human malaria 
by chemoattenuated PfSPZ vaccine. Nature. 2017 Feb 23;542(7642):445-449. 
20. Murphy SC, Duke E, Shipman KJ, et al. A randomized trial evaluating the 
prophylactic activity of DSM265 against pre-erythrocytic Plasmodium falciparum 
controlled human malaria infection by mosquito bites and direct venous 
inoculation. J Inf Dis 2018; 217: 693-702.
21. Piepoli MF, Hoes AW, Agewall S, et al. European Guidelines on cardiovascular 
disease prevention in clinical practice: The Sixth Joint Task Force of the 
European Society of Cardiology and Other Societies on Cardiovascular Disease 
Prevention in Clinical Practice (constituted by representatives of 10 societies and 
by invited experts). Developed with the special contribution of the European 
Association for Cardiovascular Prevention & Rehabilitation (EACPR). Eur Heart 
J. 2016;37(29):2315–2381.
22. Riamet Summary of Product Characteristics (May 2017). Novartis Pharma 
NV/SA. Vilvoorde, Belgium (bijsluiters.fagg-
afmps.be/registrationSearchServlet?key=BE223562&leafletType=skp). Accessed 
November 2019. 

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23. Roestenberg M, Bijker EM, Sim BK, et al. Controlled human malaria infections 
by intradermal injection of cryopreserved Plasmodium falciparum sporozoites. 
Am J Trop Med Hyg 2013 Jan;88(1): 5-13.
24. Sheehy SH, Spencer AJ, Douglas AD, et al. Optimising Controlled Human 
Malaria Infection Studies Using Cryopreserved P. falciparum Parasites 
Administered by Needle and Syringe. PLoS One. 2013 Jun 18;8(6):e65960. 
25. Shekalaghe S, Rutaihwa M, Billingsley PF, et al. Controlled human malaria 
infection of Tanzanians by intradermal injection of aseptic, purified, 
cryopreserved Plasmodium falciparum sporozoites. Am J Trop Med Hyg. 2014 
Sep;91(3):471-80.
26. Sissoko MS, Healy SA, Katile A, et al. Safety and efficacy of PfSPZ Vaccine 
against Plasmodium falciparum via direct venous inoculation in healthy malaria-
exposed adults in Mali: a randomised, double-blind phase 1 trial. Lancet Infect 
Dis. 2017 May;17(5):498-509.
27. Sulyok M, Rückle T, Roth A, et al. DSM265 for Plasmodium falciparum 
chemoprophylaxis in adults: a randomised, double blinded, Proof of concept 
Phase 1b trial with controlled human malaria infection. Lancet Infect Dis 2017; 
17: 636-644.
28. Woodrow CJ, White NJ. The clinical impact of artemisinin resistance in 
Southeast Asia and the potential for future spread. FEMS Microbiol Rev. 2017
Jan;41(1):34-48.
29. World Malaria Report 2018. World Health Organization. Geneva, Switzerland 
(http://www.who.int/malaria/publications/world-malaria-report-2018/en/). 
Accessed July 2019.

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ATTACHMENT 1: CLINICAL SCORE FOR MALARIA
Grading of the signs and symptoms specified in the table beneath according to the 
CTCAE grading scale will be performed at all relevant protocol-specified time points,
during the previous 24 h (continuously) if once daily assessments are required or during 
the previous 12 h (continuously) if assessments are performed twice daily, in accordance 
with the following:
-
Absent = 0
-
Mild = 1; equates to CTCAE grade 1
-
Moderate = 2; equates to CTCAE grade 2
-
Severe = 3; equates to CTCAE grade 3 or above
Participant ID: 
Date: 
Time of 
Onset:
Stop:
Symptom/Sign
Score (0 to 3)
qPCR
Headache
܆positive
܆negative
Myalgia (muscle ache)
Arthralgia (joint ache)
Fatigue/lethargy
Malaise (general 
discomfort/uneasiness)
Chills/Shivering/Rigors
Sweating/hot spells
Anorexia
Nausea
Vomiting
Abdominal discomfort
Fever
Tachycardia
Hypotension
TOTAL SCORE
…/42

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ATTACHMENT 2: BECK DEPRESSION INVENTORY

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ATTACHMENT 3: NORMAL RANGES FOR VITAL SIGNS 
AND ECG
NORMAL RANGES FOR VITAL SIGNS
Systolic blood 
pressure (mmHg)
Diastolic blood 
pressure (mmHg)
Pulse rate
(bpm)
Oral temperature 
(°C)
6%3
'%3
SXOVH
W
These normal ranges are applicable in supine and sitting position (after at least 10 min) 
and in the standing position (after at least 2 min in the standing position).
NORMAL RANGES FOR ECG PARAMETERS
PR
(ms)
QRS
(ms)
QTc F
(ms)
Heart rate
(bpm)
120d PR d220
QRS d120
QTc d450 for males, d470
for females.
+5
These normal ranges are applicable in supine and sitting position (after at least 10 min).

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ATTACHMENT 4: CLINICALLY ACCEPTABLE RANGES 
FOR CLINICALLY IMPORTANT STUDY INCLUSION
LABORATORY TESTS
Test
Unit
Acceptable Inclusion Range
Low
High
Sodium
mmol/L
130
150
Potassium
mmol/L
3.0
5.5
Chloride
mmol/L
85
120
Calcium (Corrected)
mg/dL
0.95 x LLN 
1.05 x ULN 
Phosphate
mmol/l
0.95 x LLN 
1.05 x ULN 
Bicarbonate
mmol/l
0.95 x LLN
1.05 x ULN
Glucose Fasted
mg/dL
N/A
1.0 x ULN 
Urea
mg/dL
N/A
1.75 x ULN
Uric acid
mg/dL
N/A
1.75 x ULN
Creatinine
mg/dL
N/A
1.0 x ULN 
Creatine kinase
U/L
N/A
< 2.5 x ULN
*eGFR
mL/min/1.73m2
60
N/A
Total Protein
g/L
[//1
[8/1
Albumin
g/L
[//1
[8/1
Total Bilirubin
mg/dL
N/A
1.25 x ULN
Direct Bilirubin
mg/dL
N/A
1.25x ULN
ALP
U/L
N/A 
1.5 x ULN
AST
U/L
N/A
1 x ULN
ALT
U/L
N/A
1 x ULN
GGT
U/L
N/A
1.5 x ULN
Lactate Dehydrogenase
U/L
0.9x LLN
1.1 x ULN
Prothrombin time INR
INR
1.0 x LLN 
1.0 x ULN 
Cholesterol
mg/dL
N/A
1.2 x ULN 

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Test
Unit
Acceptable Inclusion Range
Low
High
HDL Cholesterol
mg/dL
0.9x LLN 
N/A
LDL Cholesterol
mg/dL
N/A
1.25 x ULN 
Haemoglobin
g/dL
0.9x LLN
1.1.x ULN
Platelets 
10E9/L
0.9x LLN 
1.1 x ULN 
White Blood Cells
10E9/L
0.9x LLN 
1.1 x ULN 
Neutrophils 
10E9/L
1.0 x LLN 
1.0x ULN 
Lymphocytes
10E9/L
1.1 x LLN 
1.1 x ULN 
Monocytes
10E9/L
N/A
1.2 x ULN 
Eosinophils
10E9/L
N/A
1.0 x ULN 
Basophils
10E9/L
N/A
2.0 x ULN 
C-reactive protein (CRP)
mg/L
N/A
1.0 x ULN
Troponin-T 
(high 
sensitivity)
ng/L
N/A
< 12
Protein (dipstick)
N/A
Trace
Ketones (dipstick)
N/A
Positive
Red Blood Cells (MCS)
μL
N/A
<19
White Blood Cells (MCS)
μL
N/A
<15
Hyaline Casts (MCS)
N/A
1+