Conditions: Malaria,Falciparum Malaria Malaria Recrudescence Parasitemia Protozoan Infections Parasitic Disease
Keywords: Malaria Malaria,Falciparum Malaria Recrudescence Parasitaemia Pyronaridine
Allocation: RANDOMIZED | Intervention model: SEQUENTIAL
Enrollment: 10
Healthy volunteers: yes

Brief summary:
This is an open-label, adaptive study that will utilise the P. falciparum induced blood stage malaria (IBSM) model to characterise the pharmacokinetic/pharmacodynamic (PK/PD) profile of pyronaridine. Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this study, in cohorts of up to six participants each. Following a screening period of up to 28 days, cohorts of up to 6 healthy participants will be enrolled. Each participant will be inoculated intravenously on Day 0 with P. falciparum infected erythrocytes. Participants will be followed up daily on Days 1 to 3, and will attend the clinical unit once on Days 4, 5, 6 and 7 for clinical evaluation and blood sampling. Participants will be admitted to the clinical trial unit on Day 8 for a single oral dose of pyronaridine.

Different doses of pyronaridine will be administered across and within cohorts. Participants will be randomised to a dose group on the day of dosing. The highest dose of pyronaridine administered will be no more than 720 mg; the lowest dose administered will be no less than 180 mg.

Each subsequent cohort will be composed of up to 3 dose groups. The Safety Data Review Team (SDRT) will review all available safety and tolerability data from the previous cohort/s prior to inoculation of the next cohort.

Participants will be confined in the clinical unit for at least 96 h (Days 8 - 12) to monitor the safety and tolerability of pyronaridine dosing. Upon discharge from the clinical unit participants will be monitored on an outpatient basis up to Day 50±2. Participants will receive compulsory antimalarial rescue treatment with Riamet® (artemether/lumefantrine) on Day 47±2 or earlier.

Detailed description:
This is an open-label, adaptive study that will utilise the P. falciparum induced blood stage malaria (IBSM) model to characterise the pharmacokinetic/pharmacodynamic (PK/PD) profile of pyronaridine.

Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this study, in cohorts of up to six participants each. Following a screening period of up to 28 days, cohorts of up to 6 healthy participants will be enrolled. Each participant will be inoculated intravenously on Day 0 with approximately 2,800 viable P. falciparum infected erythrocytes. Participants will be followed up daily via phone call or text message on Days 1 to 3 post-inoculation to solicit any adverse events. Participants will attend the clinical unit once on Days 4, 5, 6 and 7 for clinical evaluation and blood sampling to monitor the progression of parasitaemia, using quantitative polymerase chain reaction (qPCR) targeting the gene encoding 18S rRNA (referred to hereafter as malaria 18S qPCR). Participants will have a nasopharyngeal aspirate (NPA) for SARS-CoV-2 on Day 6 am in the event that there is community transmission in South East Queensland of SARS-CoV-2. Participants will also have blood collected on Day 7 am to monitor haematology and biochemistry. Participants will be admitted to the clinical trial unit on Day 8 for dosing with the investigational medicinal product (IMP; pyronaridine) when parasitaemia for the majority of participants is expected to be above 5,000 parasites/mL. Pyronaridine will be administered as a single oral dose. Different doses of pyronaridine will be administered across and within cohorts in order to effectively characterise the PK/PD relationship. Each cohort will comprise up to three dose groups, with participants randomised to a dose group on the day of dosing. The highest dose of pyronaridine administered will be no more than 720 mg; the lowest dose administered will be no less than 180 mg. The proposed dosing regimen for the Cohort 1, assuming six participants are enrolled, is as below. If less than six participants are enrolled the Safety Data Review Team (SDRT) will meet between Day 0 and Day 8, to decide on the dose/s to be administered.

Dose group

1A (n=2) 360 mg

1B (n=2) 540 mg

1C (n=2) 720 mg

Each subsequent cohort will be composed of up to 3 dose groups. If more than one dose is tested in a given cohort, subjects will be randomised after inoculation day (Day 0) but prior to Day 8. The dose/s administered to Cohort 2 and Cohort 3 will be selected based on PK/PD and safety data from the preceding cohort/s. The SDRT will review all available safety and tolerability data and PK/PD analysis outcomes from the previous cohort/s prior to inoculation of the next cohort. The study will conclude when sufficient data have been obtained to define the PK/PD parameters for pyronaridine (the primary endpoint).

A maximum sample size of 18 participants administered pyronaridine is expected to be sufficient to achieve the primary endpoint.

Participants will be confined in the clinical unit for at least 96 h (Days 8 - 12) to monitor the safety and tolerability of pyronaridine dosing and to ensure adequate clinical response against P. falciparum. During confinement, regular safety assessments will be performed and blood will be collected to monitor parasite clearance and pyronaridine concentration. If participants are clinically well at the end of the confinement period, they will be discharged and monitored on an outpatient basis up to Day 50±2.

Participants will receive compulsory antimalarial rescue treatment with Riamet® (artemether/lumefantrine) on Day 47±2 or earlier.

Arms:
arm 1: Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this study, in cohorts of up to six participants each. on Day 0 participants will be inoculated intravenously with approximately 2,800 viable P. falciparum-infected erythrocytes.

Day 8 participants will be dosed with pyronaridine when parasitaemia for the majority of participants is expected to be above 5,000 parasites/mL.

Different doses of pyronaridine will be administered across and within cohorts in order to effectively characterise the PK/PD relationship.

Regular safety assessments will be performed and blood will be collected to monitor parasite clearance and pyronaridine concentration.

Participants will receive compulsory antimalarial rescue treatment with Riamet® (artemether/lumefantrine) on Day 47±2, or earlier. arm 2: Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this study, in cohorts of up to six participants each. on Day 0 participants will be inoculated intravenously with approximately 2,800 viable P. falciparum-infected erythrocytes.

Day 8 participants will be dosed with pyronaridine when parasitaemia for the majority of participants is expected to be above 5,000 parasites/mL.

Different doses of pyronaridine will be administered across and within cohorts in order to effectively characterise the PK/PD relationship.

Regular safety assessments will be performed and blood will be collected to monitor parasite clearance and pyronaridine concentration.

Participants will receive compulsory antimalarial rescue treatment with Riamet® (artemether/lumefantrine) on Day 47±2, or earlier. arm 3: Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this study, in cohorts of up to six participants each. on Day 0 participants will be inoculated intravenously with approximately 2,800 viable P. falciparum-infected erythrocytes.

Day 8 participants will be dosed with pyronaridine when parasitaemia for the majority of participants is expected to be above 5,000 parasites/mL.

Different doses of pyronaridine will be administered across and within cohorts in order to effectively characterise the PK/PD relationship.

Regular safety assessments will be performed and blood will be collected to monitor parasite clearance and pyronaridine concentration.

Participants will receive compulsory antimalarial rescue treatment with Riamet® (artemether/lumefantrine) on Day 47±2, or earlier. arm 4: This inoculated participant was treated with rescue medication instead of pyronaridine due to testing positive for SARS-CoV-2 (COVID-19).

Ten (10) participants were inoculated with P. falciparum and were included in the Full Analysis Set (FAS). Nine (9) inoculated participants were subsequently administered pyronaridine, and were included in the IMP Set, PK Analysis Set, PD Analysis Set, and the PK/PD Analysis Set. Some of the 18S qPCR data for the inoculated participant who was not administered pyronaridine was used in the PD model for PK/PD analyses.

Interventions: intervention 1: Pyronaridine intervention 2: P. falciparum IBSM infection
Intervention descriptions: intervention 1: Single dose intervention 2: Induced Blood Stage Malaria

Protocol document:
Prot_000.pdf: 
 
 
 
 
 
 
A phase 1b study to characterise the pharmacokinetic/pharmacodynamic relationship of 
pyronaridine in healthy adult participants experimentally infected with blood stage 
Plasmodium falciparum. 
 
Protocol Identifying Number: MMV_Pyronaridine_21_01 
 
Principal Investigator: Associate Professor Bridget Barber 
  
Sponsor: Medicines for Malaria Venture  
Local Sponsor: Southern Star Research 
 
Version Number: 4.0 
 
27 July 2022 
 
CONFIDENTIALITY STATEMENT 
This document contains information that is privileged or confidential. As such, it may not be 
disclosed unless specific prior permission is granted in writing by Medicines for Malaria 
Venture (MMV) or such disclosure is required by federal or other laws or regulations. Persons 
to whom any of this information is to be disclosed must first be informed that the information 
is confidential. These restrictions on disclosure will apply equally to all future information 
supplied, which is indicated as privileged or confidential. 


  
 
 
 
Table of Contents 
STATEMENT OF COMPLIANCE ............................................................................................................................. 1 
1 
PROTOCOL SUMMARY .......................................................................................................................... 3 
1.1 
SYNOPSIS ......................................................................................................................................... 3 
1.2 
Schema ................................................................................................................................................. 8 
1.3 
SCHEDULE OF ACTIVITIES ...................................................................................................... 9 
2 
INTRODUCTION .......................................................................................................................................... 12 
2.1 
Background and Study Rationale .................................................................................................... 12 
2.2 
Risk/Benefit Assessment .................................................................................................................. 13 
2.2.1 
Known Potential Risks ................................................................................................... 13 
2.2.2 
Known Potential Benefits .............................................................................................. 17 
2.2.3 
Assessment of Potential Risks and Benefits .............................................................. 17 
3 
OBJECTIVES AND ENDPOINTS .............................................................................................................. 18 
4 
STUDY DESIGN ........................................................................................................................................... 19 
4.1 
Overall Design .................................................................................................................................... 19 
4.2 
Scientific Rationale for Study Design.............................................................................................. 20 
4.3 
Justification for Dose ......................................................................................................................... 21 
4.4 
End of Study Definition ..................................................................................................................... 22 
5 
STUDY POPULATION ................................................................................................................................ 23 
5.1 
Inclusion Criteria ................................................................................................................................ 23 
5.2 
Exclusion Criteria ............................................................................................................................... 25 
5.3 
Lifestyle Considerations .................................................................................................................... 28 
5.4 
Screen Failures .................................................................................................................................. 28 
5.5 
Strategies for Recruitment and Retention ...................................................................................... 29 
6 
STUDY INTERVENTIONS .......................................................................................................................... 29 
6.1 
Study Interventions Administration.................................................................................................. 29 
6.1.1 
Study Interventions Description ................................................................................... 29 
6.1.2 
Dosing and Administration ............................................................................................ 30 
6.2 
Preparation/Handling/Storage/Accountability ................................................................................ 32 
6.2.1 
Acquisition and accountability ...................................................................................... 32 
6.2.2 
Formulation, Appearance, Packaging, and Labeling ................................................ 33 
6.2.3 
Product Storage and Stability ....................................................................................... 33 
6.2.4 
Preparation ...................................................................................................................... 34 
6.3 
Measures to Minimize Bias: Randomisation .................................................................................. 34 
6.4 
Study Interventions Compliance ...................................................................................................... 34 
6.5 
Concomitant Therapy ........................................................................................................................ 35 
6.5.1 
Rescue Medicine ............................................................................................................ 36 
7 
STUDY INTERVENTIONS DISCONTINUATION AND PARTICIPANT 
DISCONTINUATION/WITHDRAWAL .................................................................................................................... 36 
7.1 
Discontinuation of Study Interventions ........................................................................................... 36 
7.1.1 
Criteria for Administration of Rescue Medication instead of IMP dosing on Day 8
 
36 
7.2 
Volunteer Discontinuation/Withdrawal from the Study ................................................................. 37 
7.3 
Lost to Follow-Up ............................................................................................................................... 38 
8 
STUDY ASSESSMENTS AND PROCEDURES ..................................................................................... 38 
8.1 
Study Conduct Schedule .................................................................................................................. 38 
8.1.1 
Screening Visit (Day -28 to Day -1) ............................................................................. 38 


  
 
 
 
8.1.2 
Eligibility Confirmation Visit (Day -3 to Day -1) .......................................................... 40 
8.1.3 
Malaria Challenge Agent Administration (Day 0) ...................................................... 40 
8.1.4 
Outpatient Monitoring Pre-IMP Administration (Day 1 to Day 7) ............................ 41 
8.1.5 
Inpatient Observation and IMP Administration (Day 8 to Day 12) .......................... 41 
8.1.6 
Outpatient Monitoring Post-IMP Administration (Day 13 to Day 50±2) .................. 43 
8.1.7 
Antimalarial Rescue Treatments .................................................................................. 44 
8.1.8 
End of Study Visit (Day 50±2) ...................................................................................... 45 
8.2 
Pharmacokinetic and Pharmacodynamic Assessments .............................................................. 45 
8.2.1 
Pharmacokinetics ........................................................................................................... 45 
8.2.2 
Pharmacodynamics ....................................................................................................... 46 
8.3 
Safety and Other Assessments ....................................................................................................... 47 
8.3.1 
Beck Depression Inventory ........................................................................................... 47 
8.3.2 
Physical Examination .................................................................................................... 48 
8.3.3 
Body Measurements ...................................................................................................... 48 
8.3.4 
Vital signs ........................................................................................................................ 48 
8.3.5 
12-Lead Electrocardiogram (ECG) .............................................................................. 49 
8.3.6 
Laboratory Tests ............................................................................................................ 49 
8.3.7 
SARS-CoV-2 Risk Management: ................................................................................. 53 
8.3.8 
Safety Serum Retention Sample ................................................................................. 53 
8.3.9 
Malaria Future Research .............................................................................................. 54 
8.4 
Allowed Time Windows ..................................................................................................................... 54 
8.5 
Adverse Events and Serious Adverse Events ............................................................................... 55 
8.5.1 
Definition of Adverse Events (AE) ............................................................................... 55 
8.5.2 
Definition of Serious Adverse Events (SAE) .............................................................. 55 
8.5.3 
Definition of Events of Special Interest ....................................................................... 56 
8.5.4 
Classification of an Adverse Event .............................................................................. 58 
8.5.5 
Time Period and Frequency for Event Assessment and Follow-Up ....................... 59 
8.5.6 
Adverse Event Reporting .............................................................................................. 60 
8.5.7 
Serious Adverse Event and Event of Special Interest Reporting ............................ 60 
8.5.8 
Reporting Events to Participants ................................................................................. 61 
8.5.9 
Reporting of Pregnancy ................................................................................................ 61 
9 
STATISTICAL CONSIDERATIONS .......................................................................................................... 62 
9.1 
Statistical Hypotheses ....................................................................................................................... 62 
9.2 
Sample Size Determination .............................................................................................................. 62 
9.3 
Populations for Analyses .................................................................................................................. 62 
9.4 
Statistical Analyses............................................................................................................................ 62 
9.4.1 
General Approach .......................................................................................................... 62 
9.4.2 
Analysis of the Primary Endpoint ................................................................................. 63 
9.4.3 
Analysis of the Secondary Endpoints ......................................................................... 64 
9.4.4 
Safety Analyses .............................................................................................................. 65 
9.4.5 
Baseline Descriptive Statistics ..................................................................................... 66 
9.4.6 
Planned Interim Analyses ............................................................................................. 66 
9.4.7 
Sub-Group Analyses ..................................................................................................... 66 
9.4.8 
Tabulation of Individual participant Data .................................................................... 67 
9.4.9 
Exploratory Analyses ..................................................................................................... 67 
10 
SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS .............................. 67 
10.1 
Regulatory, Ethical, and Study Oversight Considerations ........................................................... 67 


  
 
 
 
10.1.1 
ETHICAL STANDARD .............................................................................................. 67 
10.1.2 
ETHICAL REVIEW .................................................................................................... 67 
10.1.3 
Informed Consent Process ........................................................................................... 68 
10.1.4 
Study Discontinuation and Closure ............................................................................. 69 
10.1.5 
Confidentiality and Privacy ........................................................................................... 69 
10.1.6 
Future Use of Stored Specimens and Data ............................................................... 70 
10.1.7 
Key Roles and Study Governance .............................................................................. 70 
10.1.8 
Safety Oversight ............................................................................................................. 72 
10.1.9 
Clinical Monitoring .......................................................................................................... 73 
10.1.10 
Quality Assurance and Quality Control ....................................................................... 74 
10.1.11 
Data Handling and Record Keeping ............................................................................ 74 
10.1.12 
Protocol Deviations ........................................................................................................ 75 
10.1.13 
Publication and Data Sharing Policy ........................................................................... 76 
10.1.14 
Conflict of Interest Policy .............................................................................................. 77 
10.2 
LIABILITY/INDEMNITY/INSURANCE .................................................................................... 77 
10.3 
Abbreviations ...................................................................................................................................... 78 
11 
REFERENCES ............................................................................................................................................. 80 
12 
APPENDIX .................................................................................................................................................... 82 


Pyronaridine VIS Protocol Version 4.0; 27 July 2022 
 
 
 
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STATEMENT OF COMPLIANCE 
Investigator declaration 
I have read the protocol and agree that it contains all necessary details for carrying out the study as 
described. I will conduct this protocol as outlined herein and will make a reasonable effort to complete 
the study within the time designated. 
I agree to personally conduct or supervise the described study. 
The study will be conducted in accordance with the following: 
 
World Medical Association Declaration of Helsinki – Ethical Principles for Medical Research 
Involving Human Participants (Fortaleza, Brazil 2013) 
 
NHMRC National Statement on Ethical Conduct in Human Research (2007, updated 2018)  
 
Integrated addendum to ICH E6(R1): Guideline for Good Clinical Practice ICH E6(R2) (November 
2016) – with introductory comments of the Australian Therapeutic Goods Administration  
 
Current ethics approved Clinical Trial Protocol. 
I agree to inform all participants that the study agents are being used for investigational purposes and I 
will ensure that the requirements related to obtaining informed consent are in accordance with 
International Council of Harmonisation (ICH) Guidelines for Good Clinical Practice (GCP) section 4.8 and 
local requirements. 
I agree to report adverse events that occur in the course of the study to the Sponsor in accordance with 
ICH Guidelines for GCP section 4.11 and local requirements. 
I have read and understand the information in the Investigator’s Brochure(s), including the potential risks 
and side effects of the study drug. 
I agree to promptly report to the Human Research Ethics Committee (HREC) all changes in the research 
activity and all unanticipated problems involving risk to participants. I will not make any changes to the 
conduct of the study without HREC and Sponsor approval, except when necessary to eliminate apparent 
immediate harm to participants. 
I agree to maintain adequate and accurate records and make those records available in accordance with 
ICH Guidelines for GCP section 4.11 and local requirements. 
I agree to ensure that all associates, colleagues, and employees assisting in the conduct of the study are 
informed about their obligations in meeting the above commitments. 
I understand that the study may be terminated or enrolment suspended at any time by the Sponsor, with 
or without cause, or by me if it becomes necessary to protect the best interest of the participants. 
 
______________________________                            Date: _____________________________ 
 Principal Investigator 


Pyronaridine VIS Protocol Version 4.0; 27 July 2022 
 
 
 
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Signatories 
The undersigned parties agree, that the protocol was written in accordance with the World Medical 
Association Declaration of Helsinki – Ethical Principles for Medical Research Involving Human Participants 
(Fortaleza, Brazil 2013), the NHMRC National Statement on Ethical Conduct in Human Research (2007, 
updated 2018), and the Integrated Addendum to ICH E6 (R1): Guideline for Good Clinical Practice E6 (R2) 
(November 2016) – with introductory comments of the Australian Therapeutic Goods Administration. 
This clinical trial protocol has been reviewed and approved by the Sponsor. 
Name  
Signature  
Date  
Medical Director:  
Anne Claire Marrast, MD  
Medicines for Malaria Venture  
 
 
Vice President, Head of Integrated Sciences:  
Jörg Moehrle, Associate Professor  
Medicines for Malaria Venture 
 
 
 
 


Pyronaridine VIS Protocol Version 4.0; 27 July 2022 
 
 
 
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1 
 
PROTOCOL SUMMARY 
1.1 
SYNOPSIS  
 
Title: 
A phase 1b study to characterise the pharmacokinetic/pharmacodynamic relationship of 
pyronaridine in healthy adult participants experimentally infected with blood stage 
Plasmodium falciparum.  
 
Study 
Description: 
This is an open-label, adaptive study that will utilise the P. falciparum induced blood 
stage malaria (IBSM) model to characterise the pharmacokinetic/pharmacodynamic 
(PK/PD) profile of pyronaridine.   
Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this 
study, in cohorts of up to six participants each. Following a screening period of up to 28 
days, cohorts of up to 6 healthy participants will be enrolled. After confirmation of 
eligibility, including a Rapid Antigen Test (RAT) for SARS-CoV-2, each participant will be 
inoculated intravenously on Day 0 with approximately 2,800 viable P. falciparum-
infected erythrocytes. Participants will be followed up daily via phone call or text 
message on Days 1 to 3 post-inoculation to solicit any adverse events. 
 
Participants will attend the clinical unit once on Days 4, 5, 6 and 7 for clinical evaluation 
and blood sampling to monitor the progression of parasitaemia, using quantitative 
polymerase chain reaction (qPCR) targeting the gene encoding 18S rRNA (referred to 
hereafter as malaria 18S qPCR). Participants will have a nasopharyngeal aspirate (NPA) 
for SARS-CoV-2 on Day 6 am. Participants will also have blood collected on Day 7 am to 
monitor haematology and biochemistry.  
 
Participants will be admitted to the clinical trial unit on Day 8 for dosing with the 
investigational medicinal product (IMP; pyronaridine) when parasitaemia for the 
majority of participants is expected to be above 5,000 parasites/mL. Participants will 
need to return a negative (RAT) for SARS-CoV-2 prior to admission. 
Pyronaridine will be administered as a single oral dose. Different doses of pyronaridine 
will be administered across and within cohorts in order to effectively characterise the 
PK/PD relationship. Each cohort will comprise up to three dose groups, with participants 
randomised to a dose group on the day of dosing. The highest dose of pyronaridine 
administered will be no more than 720 mg; the lowest dose administered will be no less 
than 180 mg. The proposed dosing regimen for the Cohort 1, assuming six participants 
are enrolled, is as below. If less than six participants are enrolled the Safety Data Review 


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Team (SDRT) will meet between Day 0 and Day 8, to decide on the dose/s to be 
administered. 
 
Dose group 
 
1A (n=2) 
1B (n=2) 
1C (n=2) 
Pyronaridine 
360 mg  
540 mg 
720 mg  
Each subsequent cohort will be composed of up to 3 dose groups. If more than one dose 
is tested in a given cohort, subjects will be randomised after inoculation day (Day 0) but 
prior to Day 8. The dose/s administered to Cohort 2 and Cohort 3 will be selected based 
on PK/PD and safety data from the preceding cohort/s. The SDRT will review all available 
safety and tolerability data and PK/PD analysis outcomes from the previous cohort/s 
prior to inoculation of the next cohort. The study will conclude when sufficient data have 
been obtained to define the PK/PD parameters for pyronaridine (the primary endpoint).   
A maximum sample size of 18 participants administered pyronaridine is expected to be 
sufficient to achieve the primary endpoint. 
Participants will be confined in the clinical unit for at least 96 h (Days 8 - 12) to monitor 
the safety and tolerability of pyronaridine dosing and to ensure adequate clinical 
response against P. falciparum. During confinement, regular safety assessments will be 
performed and blood will be collected to monitor parasite clearance and pyronaridine 
concentration. If participants are clinically well at the end of the confinement period, 
they will be discharged and monitored on an outpatient basis up to Day 50±2.  
Participants will receive compulsory antimalarial rescue treatment with Riamet® 
(artemether/lumefantrine) on Day 47±2 or earlier in the following cases: 
 
Parasitaemia is not reduced by approximately 10-fold at the end of the 96 
hour confinement period (Day 12) when compared with peak parasitaemia. 
 
Parasite regrowth occurs after an initial reduction in parasitaemia, with 
regrowth defined as ≥5000 parasites/mL and a two-fold parasitaemia 
increase within 48 hours.  
 
Volunteer discontinuation/withdrawal from the study. 
 
Investigator’s discretion in the interest of participant safety. 
Parasite lifecycle stages (including gametocytes) will be monitored at select time points 
during the study by collecting blood samples and using quantitative reverse-
transcriptase polymerase chain reaction (qRT-PCR) targeting parasite lifecycle stage-
specific mRNA transcripts. 
If gametocytes are present as determined by qRT-PCR at any time point after the 
confinement period, a single dose of Primacin® will be administered at the Investigator’s 
discretion. If an allergy or contraindication to Riamet® develops, Malarone® 


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(atovaquone/proguanil) will be administered. If in the rare event participants are unable 
to complete oral antimalarial rescue treatment, they will be admitted to hospital to 
receive intravenous artesunate. 
Objectives: 
 
Primary Objective: 
 
To characterise the PK/PD relationship of pyronaridine in healthy participants 
experimentally infected with blood stage P. falciparum. 
Secondary Objectives: 
 
To evaluate the safety and tolerability of single oral doses of pyronaridine in 
healthy participants experimentally infected with blood stage P. falciparum. 
 
To characterise the parasite clearance kinetics following single doses of 
pyronaridine in healthy participants experimentally infected with blood-stage P. 
falciparum. 
 
To characterise the pharmacokinetics of pyronaridine following single oral dose 
administration in healthy participants experimentally infected with blood stage 
P. falciparum. 
 
To characterise the effect of pyronaridine on P. falciparum 3D7 parasite viability 
using phenotypic and molecular approaches. 
Endpoints: 
Primary Endpoint: 
 
The PK/PD relationship between pyronaridine blood concentrations and blood 
stage asexual parasitaemia. 
  
Secondary Endpoints: 
 
The safety and tolerability of pyronaridine will be determined by recording the 
incidence, severity and relationship to pyronaridine of adverse events (as 
determined by clinical, laboratory and ECG examinations) from Day 8 up to Day 
50±2. 
 
The parasite clearance kinetics following dosing with pyronaridine will be 
determined by calculating the parasite reduction ratio over a 48 hour period 
(PRR48) and corresponding parasite clearance half-life (PCt1/2). 
 
The pharmacokinetics of pyronaridine will be determined by calculating the 
following parameters using non-compartmental methods: Cmax, Tmax, tlag, AUC0-
last, AUC0-∞, CL/F, Vz/F, λz, t1/2.  
 
Parasite growth in ex vivo cultures. 
 Population: 
Participants will be malaria-naïve healthy males and females, aged between 18-55 years 
old, who meet all the inclusion criteria and none of the exclusion criteria. Women of 
childbearing potential (WOCBP) willing to use highly effective double barrier 
contraception will be eligible. Efforts will be made to ensure a reasonable gender 


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balance. Up to 18 participants will be enrolled in cohorts of up to 6 participants, with 
each cohort comprising up to 3 dose groups.  
Phase: 
Phase 1b   
Sites 
enrolling 
participants: 
The trial is planned to be performed at the University of Sunshine Coast Clinical Trials 
Centres in Queensland, Australia.  
Moreton Bay  
Health Hub Morayfield 19-31 Dickson Road Morayfield QLD 4506  
South Bank 
Building A2, SW1 Complex 52 Merivale Street South Brisbane QLD 4101 
Description of 
Study 
Interventions: 
Investigational Medicinal Product: 
Pyronaridine tablets (each containing 180 mg pyronaridine tetraphosphate) will be 
manufactured by Shin Poong Pharmaceutical Co., Ltd (Seoul, South Korea). Each 
participant will be administered the appropriate dose of tablets orally at the clinical trial 
unit under direct staff observation. Pyronaridine will be administered after a ≥ 8 hour 
fast.  
Malaria Challenge Agent: 
The P. falciparum 3D7 master cell bank (MCB) was produced from blood collected from 
a donor with clinical manifestation of malaria. Each challenge dose will be prepared 
aseptically at Q-Gen Cell Therapeutics (QIMR Berghofer) from an aliquot of the P. 
falciparum 3D7 MCB. Each participant will be inoculated intravenously with a dose of 
approximately 2,800 viable P. falciparum 3D7-infected erythrocytes in 2 mL of saline for 
injection. 
Antimalarial Medications: 
Compulsory 
Riamet® (each tablet containing 20 mg artemether and 120 mg lumefantrine) is 
marketed by Novartis Pharmaceutical Australia. A standard treatment course comprises 
6 doses of 4 tablets administered orally over 60 hours (total course of 24 tablets). Each 
dose of tablets should be taken with food or drinks rich in fat (e.g. milk). 
 
If required 
 
Primacin® (each tablet containing 13.2 mg primaquine phosphate equivalent to 7.5 
mg of primaquine) is marketed by Boucher & Muir Pty Ltd. If gametocytes are 
present as determined by qRT-PCR at any time point after the confinement period, 
a single dose of Primacin® will be administered at the Investigator’s discretion. A 
standard treatment course comprises a single dose of 6 tablets administered orally, 
unless G6PD deficient in which case a low dose of 15 mg will be administered. 


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 
Malarone® (each tablet containing 250 mg atovaquone and 100 mg proguanil 
hydrochloride) is marketed by GlaxoSmithKline Australia Pty Ltd. A standard 
treatment course comprises three doses of 4 tablets administered orally (one dose 
daily for 3 days; total course of 12 tablets). 
 
Intravenous artesunate is the recommended parenteral treatment for malaria in 
Australia. The recommended dose regimen is 2.4 mg/kg at approximately 0, 12, 24, 
48 hours and then daily for up to 7 days or until able to take oral drugs. 
Study 
Duration: 
It is estimated that the clinical portion of the study will be completed in 12 months. 
Volunteer 
Duration: 
Approximately 79 days for each participant which includes a screening period (up to 28 
days), a period of observation following malaria challenge (8 days), follow-up after 
administration of the IMP (39 days) and follow up after compulsory antimalarial rescue 
treatment (3 days). 


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1.2 
SCHEMA 
Malaria challenge 
agent administration 
Day -28 
                Day 0                                   Day 8         Day 12                            Day 50±2  
Pyronaridine 
dosing 
Screening 
Outpatient  
malaria  
monitoring 
Clinic 
confinement 
Outpatient 
follow-up 
End of 
study 
     Antimalarial rescue 
treatment (Day 47±2 
or earlier if required) 
   Study 
Timeline  


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9 
 
1.3 
SCHEDULE OF ACTIVITIES 
 
The below table summarises the activities and procedures to be conducted as per this protocol during screening, eligibility, malaria inoculation, 
out patient monitoring, confinement, and at the end of study. This is a summary only, Section 8.1 provides detailed information about the 
assessments required at each time-point. There may be multiple samples/assessments for some activities on a particular day. Sections 8.2-8.3 
provide more information about how to perform the assessments.  
 
Screening 
Eligibilitya 
Malaria 
inoculation 
Phone 
contact 
Malaria 
monitoringb 
IMP administration and 
clinical unit confinementc 
Out-patient 
monitoringd 
EOS 
Day 
-28 to -1 
-3 to -1 
0 
1 to 3 
4 to 7 
8 
9 
10 
11 
12 
13 to 50±2 
50±2 
Eligibility assessments 
Informed consent 
X 
 
 
 
 
 
 
 
 
 
 
 
Beck Depression Inventory-
II 
X 
 
 
 
 
 
 
 
 
 
 
 
Cardiovascular risk 
X 
 
 
 
 
 
 
 
 
 
 
 
Demography 
X 
 
 
 
 
 
 
 
 
 
 
 
Medical and social history, 
inc/exc. criteria, & prior 
medications 
X 
 
X 
 
 
 
 
 
 
 
 
 
Drug & alcohol screen 
X 
 
X 
 
 
X 
 
 
 
 
 
 
Body height 
X 
 
 
 
 
 
 
 
 
 
 
 
Body weight 
X 
 
 
 
 
 
 
 
 
 
 
X 
Serology & RBC 
alloantibody 
X 
 
 
 
 
 
 
 
 
 
 
X 
Coagulation profile 
X 
 
 
 
 
 
 
 
 
 
 
 
COVID-19 testingl 
 
X 
X 
 
X (D 6 am) 
X 
 
 
 
 
 
 
G6PD 
X 
 
 
 
 
 
 
 
 
 
 
 
Serum pregnancy test (all 
females at screening & 
WOCBP at EOS) 
X 
 
 
 
 
 
 
 
 
 
 
X 
Urine pregnancy test 
(WOCBP) 
 
 
X 
 
 
X 
 
 
 
 
 
 


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10 
 
 
Screening 
Eligibilitya 
Malaria 
inoculation 
Phone 
contact 
Malaria 
monitoringb 
IMP administration and 
clinical unit confinementc 
Out-patient 
monitoringd 
EOS 
Day 
-28 to -1 
-3 to -1 
0 
1 to 3 
4 to 7 
8 
9 
10 
11 
12 
13 to 50±2 
50±2 
FSH (post-menopausal 
women)  
X 
 
 
 
 
 
 
 
 
 
 
 
Safety assessments 
Full physical exam 
X 
 
 
 
 
 
 
 
 
 
 
X 
Abbreviated physical exam 
 
 
X 
 
 
X 
 
 
 
 
 
 
Symptom-directed physical 
exame 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
 
Vital signsf 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
ECG 
X 
 
X 
 
 
X 
X 
X 
X 
X 
X 
X 
Urinalysis 
X 
X 
 
 
 
 
 
 
 
 
 
X 
Haem. & Biochem.  
X g 
X 
 
 
X (D 7 am) 
X 
 
 
X 
 
X 
X 
Safety serum retention  
 
 
X 
 
 
 
 
 
 
 
 
X 
Diary card 
 
 
X 
X 
X 
 
 
 
 
 
X 
X 
AEs & Conmeds. 
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
Study interventions 
Challenge agent inoculation 
 
 
X 
 
 
 
 
 
 
 
 
 
Pyronaridine administration 
 
 
 
 
 
X 
 
 
 
 
 
 
Definitive/rescue 
antimalarial  treatment 
 
 
 
 
 
 
 
 
 
 
Xh 
 
Malaria monitoring and PK blood sampling 
Malaria 18S qPCR i 
 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
Parasite lifecycle stage qRT-
PCR blood sampling 
 
 
 
 
 
 
 
 
 
 
X 
 
Pyronaridine concentrationj  
 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
Other 
Malaria future research 
(optional) 
 
 
X 
 
 
X 
 
 
 
 
Xm 
X 
Parasite viability ex vivo 
growthk  
 
 
 
 
 
X 
X 
X 
X 
X 
X (parasite 
regrowth only) 
 
AEs: adverse events; ECG: electrocardiograph; EOS: End of Study; G6PD: glucose-6-phosphate dehydrogenase qPCR: quantitative polymerase chain reaction; 
qRT-PCR: quantitative reverse-transcriptase polymerase chain reaction; RBC: red blood cell; WOCBP: women of childbearing potential. 


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11 
 
 
a This visit is not required in the event that the screening visit is conducted within this period. 
b Daily visits. 
c See Section 8.1.5 for details of assessments to be performed prior to and after pyronaridine administration. Also note that there are multiple 
samples/assessments for some activities on the confinement days.  
d Follow up visits will occur at least three times per week. See Section 8.1.6 for the specific timings of different activities.  
e Symptom-directed physical exam will only be performed if clinically indicated. 
f Supine & standing blood pressure and heart rate required at screening; seated measurements only at other time-points. On Day 0, record vital signs prior to 
inoculation and prior to leaving the clinical trial unit (approximately 60 minutes after inoculation). Record vital signs 3 times a day whilst confined. 
g Haematology and biochemistry at screening includes lipids for cardiovascular risk factor.  
h Riamet® will be administered to all participants on Day 47±2 or earlier as outlined in Section 8.1.7. Participants may be treated with Primacin® if gametocytaemia 
is suspected from parasite lifecycle stage qRT-PCR at the Investigator’s discretion, to ensure clearance of gametocytes. If needed, participants will be administered 
a single oral dose of six Primacin® tablets (total dose 45 mg primaquine) with food, unless G6PD deficient in which case a low dose of 15 mg (two tablets) will be 
administered. 
I Blood samples for malaria 18S qPCR will be collected following pyronaridine treatment at 4, 8, 12, 16, 20, 24, 30, 36, 42, 48, 60, 72, 84 and 96 hours. As 
outpatients, blood will be collected at least three times per week until antimalarial rescue treatment. Allowed time windows for sample collection are specified 
in Section 8.4. 
j Blood samples for drug concentration measurements will be collected following pyronaridine treatment at 1, 2, 3, 4, 6, 8, 12, 16, 20, 24, 30, 36, 42, 48, 60, 72, 
84 and 96 hours. As outpatients, blood will be collected on the following days: 13, 15, 17, 20, 22, 24, 27, 29, 32, 39, 44, 47 and 50. Allowed time windows for 
sample collection are specified in Section 8.4. 
k Blood samples for parasite viability ex vivo growth will be collected following pyronaridine treatment at 4, 8, 12, 16, 20, 24, 48, 72 and 96 hours. 
l Perform SARS-CoV-2 testing at the eligibility visit (NPA PCR), or at the screening visit if this visit is conducted two days prior to inoculation, Day 0 (RAT), Day 
6am (NPA PCR) and Day 8 (RAT). Further details are specified in Section 8.3.7.  
m Blood samples for future malaria research will be collected during the outpatient monitoring phase on Days 15±1 and 29±1 
 
 


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12 
 
2 
 
INTRODUCTION 
 
2.1 
BACKGROUND AND STUDY RATIONALE  
Malaria is a mosquito-borne parasitic disease prevalent in tropical and subtropical regions around the 
world. It is caused by infection with the Plasmodium parasite of which five main species are known to 
infect humans: P. falciparum, P. vivax, P. ovale, P. malariae and P. knowlesi. Of these species, P. falciparum 
is responsible for the majority of the malaria burden, particularly in sub-Saharan Africa. The malaria 
parasite has a complex life cycle in humans consisting of both a liver stage and a blood stage. Clinical 
symptoms of malaria infection are due to the asexual blood stage of the parasite’s life cycle. Malaria is 
responsible for significant global morbidity and mortality, with an estimated 229 million new cases and 
409,000 deaths worldwide in 2019 [1]. The emergence of antimalarial drug resistance has been a major 
setback for malaria control and progress towards the goal of elimination. Resistance has emerged to all 
antimalarials in widespread use, including artemisinin-based combination therapies (ACTs), the current 
first line treatment for uncomplicated P. falciparum malaria. Although resistance to ACTs has not yet been 
reported in Africa, the emergence of such resistance would likely have devastating consequences [2]. 
Thus, the development of novel antimalarial therapies are required to control malaria in regions with high 
prevalence of artemisinin-resistance, and prevent the emergence of resistance in sub-Saharan Africa 
where the world’s malaria morbidity and mortality rates are highest. 
Pyronaridine is an antimalarial compound developed in China in the 1970’s and 1980’s and has been 
shown to have potent activity towards blood-stage P. falciparum parasites, including isolates resistant to 
chloroquine, quinine, amodiaquine, pyrimethamine or mefloquine [3]. The antimalarial activity of 
pyronaridine is thought to occur primarily due to inhibition of hemozoin formation, which results in 
accumulation of toxic hematin in the parasite food vacuole [4]. Pyronaridine has been used as a single 
agent for the treatment of malaria in China for over 30 years. It was developed as a fixed-dose 
combination therapy with artesunate (an artemisinin derivative) for the treatment of acute 
uncomplicated P. falciparum malaria and blood stage P. vivax (Pyramax®, co-developed by Medicines for 
Malaria Venture, Geneva, Switzerland and Shin Poong Pharmaceutical Co., Ltd, Seoul, South Korea). 
Pyramax® received a positive opinion by European Medicines Agency (EMA) under the article 58 process 
[evaluation of medicinal products intended exclusively for markets outside the European Community]) in 
2012 and 2015. Pyramax® was also added to the World Health Organization’s (WHO) Model List of 
Essential Medicines (EML) and Model List of Essential Medicines for Children (EMLc) in 2017 and is now 
registered in 29 countries worldwide. Its procurement and use at a country level is further supported by 
a WHO Information Note of October 2019. 
Despite the fact that pyronaridine has been used to successfully treat malaria for many years, either in 
monotherapy or in combination with other antimalarial drugs [3], its antimalarial activity in humans has 
not been completely characterised. Determining the relationship between pyronaridine pharmacokinetics 
(PK) and pharmacodynamic (PD) antimalarial activity in humans would inform efforts to broaden its use 
in new antimalarial combination therapies. Combination therapies are the focus of antimalarial drug 


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13 
 
development to reduce the risk of selecting for resistant mutants and to target multiple stages of the 
parasite lifecycle including the transmissible gametocytes [5]. Further, using a combination of two or more 
drugs is typically considered a prerequisite for achieving cure without the need for an extended dosing 
regimen, which would pose compliance challenges. In the case of ACTs, the fast acting artemisinin 
component rapidly reduces the parasite burden, while a partner drug with a more moderate rate of action 
but longer elimination half-life is relied upon to clear residual parasites and prevent recrudescence. The 
rate of action of pyronaridine in clearing malaria parasitaemia in humans is unclear; however clinical 
pharmacokinetic studies have demonstrated that it has a relatively long elimination half-life of 
approximately 14 days [3] and thus may be able to offer extended protection from recrudescence.  
Volunteer infection studies (VIS) using the induced blood stage malaria (IBSM) model have previously 
successfully characterised the PK/PD relationship of several experimental antimalarial compounds as well 
as antimalarial drugs currently in use [6-9]. The IBSM model involves intravenous inoculation of healthy 
adult participants with blood-stage parasites and administration of the test antimalarial when 
parasitaemia reaches a predefined threshold. Frequent blood sampling is performed to measure 
parasitaemia and drug concentration kinetics over the course of the study, thus providing data for PK/PD 
modelling analyses. Such VIS have been shown to accurately predict the activity of investigational 
antimalarials in studies in endemic populations [10,11].  
The current study aims to determine the PK/PD relationship of single doses of pyronaridine following 
administration in healthy adult participants using the P. falciparum IBSM model. Data obtained in this 
study will support the use of pyronaridine in new antimalarial combination therapies by informing partner 
drug selection and dosing considerations.  
2.2 
RISK/BENEFIT ASSESSMENT   
2.2.1 KNOWN POTENTIAL RISKS  
Risks associated with the Investigational Medicinal Product (pyronaridine) 
Pyronaridine in a fixed dose combination with artesunate received positive scientific opinion from EMA 
through Article 58 and is registered in many malaria endemic countries in Africa and South-East Asia as 
Pyramax® for treatment of acute uncomplicated malaria for both adults and children.  
Through the extensive use of Pyramax® in many African and South-East Asian countries, the safety and 
tolerability of pyronaridine when combined with artesunate for the treatment of uncomplicated malaria 
is  well established. 
In studies involving patients with malaria or healthy volunteers, pyronaridine and artesunate combination 
treatment was associated with transient elevations in alanine aminotransferase (ALT) in approximately 
6% of subjects, which were considered to be primarily due to pyronaridine [12]. In approximately 1.3% of 
cases these rises were >5 times the upper limit of normal (x ULN) without clinical sequelae. Transaminase 
rises occurred within the first 3-7 days and returned to within normal limits generally by Day 14. In a 


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14 
 
repeat dose study, there was no increased risk of transaminitis when the pyronaridine/artesunate 
combination was administered once, or more than once as a repeat dose [12]. In a study which included 
patients with baseline transaminase levels >2× ULN, there was no clinical evidence of hepatotoxicity. The 
early onset (Day 3-7) and rapid resolution are consistent with a direct, low-level toxicity and do not 
indicate a risk of progressive liver injury with 3-day combination treatment [12].  
Apart from the transient increase in transaminases., the most frequently reported adverse events (AEs; 
>1/100 to < 1/10) were headache, eosinophilia, neutropenia, anaemia, increased platelet count, vomiting 
(2.2 % to 2.5 %), abdominal pain, bradycardia, and hypoglycaemia [13]. Analyses performed in patients 
with malaria suggest that no clinically concerning QTc prolongation is expected at therapeutic 
concentrations of pyronaridine [14].  
Additional information on non-clinical and clinical studies conducted with pyronaridine is provided in the 
IB.  
Risks associated with the P. falciparum induced blood stage malaria model (IBSM) 
Risks pertaining to the IBSM model are development of blood borne infections, reaction to the blood 
sample, severity of malaria infection (including development of haematological adverse events such as 
neutropenia or leucopenia), development of liver function abnormalities, and occurrence of cardiac AE). 
These are each discussed in detail below. 
Risk management of blood borne infections in IBSM 
The P. falciparum 3D7 challenge agent will be used to induce blood stage malaria in this trial. Although 
the challenge agent contains a very small amount of blood, risk of a transfusion-transmissible infection in 
this trial is extremely low. Firstly, donors were screened and tested negative for the presence of active 
blood borne infections. Secondly, the Australian Red Cross Blood Service (Blood Service) removed white 
blood cells from the donor blood to lower the risk of a transfusion-transmissible infection. Thirdly, the 
volume of blood associated with the malaria challenge inoculum is many thousands of times smaller than 
in a transfused unit (i.e., a relatively lower risk of infection). As part of the safety monitoring, all 
participants will be screened for blood borne infections before and after the trial. To date, no blood borne 
infections have been reported in any of the 435 participants who have received this P. falciparum 3D7 
challenge agent in IBSM clinical trials. 
Risk management of reaction to the blood sample in IBSM 
The risk of developing red blood cell (RBC) alloantibodies and/or experiencing an acute haemolytic 
reaction in this trial is considered extremely low because the donor blood used to produce the challenge 
agent was blood group O (RhD) negative. People with this blood group are generally considered “universal 
donors”.  However, it is possible that participants could suffer a transfusion reaction after they receive 
the challenge agent, or could develop alloantibodies to the donor RBCs that may make blood transfusion 
more difficult in the future. To date, one participant has developed an antibody response to a minor Rh 
antigen (anti-E antibody) following inoculation with the P. falciparum 3D7 challenge agent. However, 


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15 
 
there was no laboratory evidence to indicate that the specific Rh phenotype of the donor RBCs in the 
challenge agent stimulated production of this anti-E alloantibody.  
Alloimmunisation has been observed in an IBSM study testing a genetically modified P. falciparum 3D7 
strain. This study investigated the safety, infectivity and immunogenicity of a strain in which the gene 
encoding the knob associated histidine rich protein (kahrp) had been deleted in order to explore its 
potential use as a live-attenuated malaria vaccine. In this study, two participants developed 
alloimmunisation (anti-C and anti-P1 antibodies in one participant and anti-c antibodies in the other 
participant) after administration of the highest dose of parasites (approximately 1000 fold higher dose of 
parasites than routinely administered in IBSM studies using the wild-type P. falciparum 3D7 strain). 
However, the total dose of erythrocytes (including non-parasitized) administered was similar to that 
routinely administered in IBSM studies (approximately 1×108 erythrocytes). Although anti-P1 antibodies 
are not considered clinically important for transfusion reactions, anti-C and anti-c antibodies may result 
in a delayed transfusion reaction characterized by slow drop in hemoglobin over two weeks post-
transfusion. Consultation with transfusion medicine specialists at the completion of the study indicated 
there was no immediate risk if these participants were to require emergency administration of unmatched 
Group O Rh (D) negative blood, and in the setting of routine blood transfusion a full cross-match would 
obviate such a reaction. Additionally, since both antibodies were of low titre, there was a possibility that 
they may diminish over time. None of the other 6 participants enrolled in this study were found to have 
developed alloimmunisation; this includes 4 participants administered a lower dose of parasites and 2 
participants administered the same dose of parasites as the 2 participants who developed 
alloimmunisation.  
Participants will be monitored for signs and symptoms in the period immediately after administration of 
the challenge agent to further assess the risk of the challenge agent causing a transfusion reaction. All 
participants will be tested for RBC alloantibodies at screening and at the end of the trial as part of safety 
monitoring.  
Women of childbearing potential (WOCBP) have a small additional risk if they develop RBC alloantibodies, 
as this could cause problems during pregnancy. WOCBP who have participated in several IBSM trials with 
P. falciparum isolate 3D7 have had no known issues to date. Including WOCBP in the trial enhances the 
generalisability of the trial results. 
Risk management of severity of malaria infection in IBSM 
The number of viable blood stage parasites that will be used to infect the participants in this trial 
(~2,800) is substantially lower than the parasitaemia induced from the bite of a single malaria-infected 
mosquito (~30,000 parasites are released into the blood when they break out of a single infected 
liver cell. In this trial, parasite growth and malaria symptoms will be closely monitored in participants 
following administration of the challenge agent. The threshold for commencement of antimalarial drug 
treatment defined for Day 8 has been selected because it is before the time-point at which clinical 
symptoms of malaria are likely to occur. 


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16 
 
Participants will be admitted to the clinical trial unit for earlier treatment if a participant experiences an 
serious adverse event (SAE)  or ≥ Common Terminology Criteria for Adverse Events (CTCAE) Grade 3 event 
deemed related to malaria and not self-resolved or relieved with concomitant medications, or the 
Investigator considers it necessary for participant safety.  Blood will be collected at specified time-points 
to monitor for haematological adverse events such as neutropenia or leucopenia that may occur as a 
result of early malaria infection.  
 
Risk management of liver function abnormalities in IBSM 
Transient, asymptomatic liver function test (LFT) abnormalities, including rare cases of alanine 
aminotransferase (ALT) and/or aspartate aminotransferase (AST) elevations >10 fold the upper limit of 
normal (×ULN), have been reported in several participants in IBSM studies. However, changes in bilirubin 
have only been reported in one participant with unappreciated pre-existing liver disease. The LFT 
abnormalities did not require treatment, and resolved by the end of the studies. A few cases of the LFT 
elevations were considered serious AEs (SAEs) by two Pharma sponsors due to internal processes for SAE 
notifications. An independent review involving drug-induced liver injury experts found these liver function 
abnormalities are most likely a direct consequence of the malaria infection rather than a direct drug-
induced liver injury caused by an investigational antimalarial drug. As a precaution, all participants with 
LFTS above ULN will be excluded in this trial. All enrolled participants will undergo regular safety 
monitoring to assess for asymptomatic liver function abnormalities. Participants will be required to 
minimise intake of possibly hepatotoxic substances, such as alcohol and paracetamol, during the trial. 
Drugs of abuse are not permitted under any circumstance.  Given the fact that pyronaridine has also been 
associated with LFT abnormalities, participants with an ALT or AST > 2 x ULN, or a total bilirubin > 1 x ULN, 
will be dosed with compulsory antimalarial treatment rather than pyronaridine (see above, “Risks 
associated with IMP [pyronaridine]”). 
Risk management of cardiac adverse events 
There have been four cardiac SAEs reported in healthy participants in the Netherlands participating in 
malaria challenge studies using sporozoites (i.e., direct feeds by infected mosquitoes rather than IBSM 
infection). These four cardiac SAEs are described in the P. falciparum 3D7 challenge agent IB; two of the 
events have also been published as case reports [20,21]. No cardiac SAEs caused by the challenge agents 
have been reported in IBSM studies. However, in a recent trial, 2 participants (one infected with the P. 
falciparum 3D7 challenge agent and one infected with another malaria challenge agent strain [P. 
falciparum K13]) developed ventricular extra systoles that were classified as moderate AEs possibly 
related to malaria. As a precaution, people at significant risk of cardiovascular disease will be excluded 
from participating in IBSM studies, and regular safety monitoring, including physical examination and ECG 
recordings, will take place for all participants. Follow-up with a cardiologist is also available if any cardiac 
AEs are seen during the IBSM studies.  
 
 


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Risks associated with antimalarial rescue medications  
Risks related to use of artemether-lumefantrine (Riamet®), primaquine phosphate (Primacin®) and 
atovaquone/proguanil (Malarone®) are detailed in the prescribing information provided by 
manufacturers. Primacin® may cause severe haemolytic anaemia in participants with glucose-6-
phosphate dehydrogenase (G6PD) deficiency. Participants will be tested for G6PD deficiency at screening 
to ensure the safety of Primacin®. Participants with a severe G6PD deficiency will not be eligible for the 
study, however those with a mild or moderate G6PD deficiency may still be enrolled. Participant’s G6PD 
status will determine how they are treated with Primacin®. A dosing table based on G6PD status will be 
provided in the Pharmacy Manual. To mitigate any other potential risks, the wellbeing of participants 
during and after administration of rescue treatment will be appropriately monitored. Participants who 
have any known contra-indication to any of the rescue medications according to the applicable labelling 
at screening will be excluded from participating in the study.   
The overall risks to the participants in this study will be managed by frequent safety monitoring, including 
close monitoring during the period of confinement at the clinical unit when a single oral dose of 
pyronaridine will be administered. Safety monitoring will include clinical laboratory safety tests 
(biochemistry and haematology), physical examination, vital signs, ECG analysis and adverse event 
monitoring. Throughout the study, the safety, parasitaemia and PK data will be assessed by the PI, Sponsor 
Medical Director and Medical Monitor and the SDRT prior to commencing subsequent cohorts. 
2.2.2 KNOWN POTENTIAL BENEFITS  
There are no expected clinical benefits for the healthy participants who will participate in this trial. There 
may be benefits to others in the future if the results of this study lead to improved treatment outcomes 
in preventing people from contracting malaria and in treating patients infected with malaria.  
 
2.2.3 ASSESSMENT OF POTENTIAL RISKS AND BENEFITS  
On the basis of the safety provisions and risk minimisation activities outlined in Section 2.2.1, the overall 
risk to the participants in this trial is considered to be minimal and acceptable, and the potential of future 
improved treatment for malaria is considered to outweigh these potential risks.  
 
 


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18 
 
 
3 
OBJECTIVES AND ENDPOINTS 
 
 
OBJECTIVES 
ENDPOINTS 
Primary 
To characterise the PK/PD relationship of 
pyronaridine 
in 
healthy 
participants 
experimentally infected with blood stage P. 
falciparum. 
The PK/PD relationship between pyronaridine 
blood concentrations and blood stage asexual 
parasitaemia.  
Secondary 
To evaluate the safety and tolerability of single 
oral 
doses 
of 
pyronaridine 
in 
healthy 
participants experimentally infected with blood 
stage P. falciparum. 
The safety and tolerability of pyronaridine will be 
determined by recording the incidence, severity 
and relationship to pyronaridine of adverse 
events (as determined by clinical, laboratory and 
ECG examinations) from Day 8 up to Day 50±2. 
To characterise the parasite clearance kinetics 
following single doses of pyronaridine in 
healthy participants experimentally infected 
with blood-stage P. falciparum. 
The parasite clearance kinetics following dosing 
with pyronaridine will be determined by 
calculating the parasite reduction ratio over a 48 
hour period (PRR48) and corresponding parasite 
clearance half-life (PCt1/2). 
To characterise the pharmacokinetics of 
pyronaridine 
following 
single 
oral 
dose 
administration 
in 
healthy 
participants 
experimentally infected with blood stage P. 
falciparum. 
The pharmacokinetics of pyronaridine will be 
determined 
by 
calculating 
the 
following 
parameters using non-compartmental methods: 
Cmax, Tmax, tlag, AUC0-last, AUC0-∞, CL/F, Vz/F, λz, t1/2. 
To characterise the effect of pyronaridine on P. 
falciparum 
3D7 
parasite 
viability 
using 
phenotypic and molecular approaches. 
Parasite growth in ex vivo cultures. 
 
 
 


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19 
 
4 
STUDY DESIGN  
 
4.1 
OVERALL DESIGN 
This is an open-label, adaptive study that will utilise the P. falciparum IBSM model to characterise the 
PK/PD profile of pyronaridine.   
Up to 18 healthy, malaria naïve adult participants are planned to be enrolled into this study, in cohorts of 
up to six participants each. Following a screening period of up to 28 days, cohorts of up to 6 healthy 
participants will be enrolled. After confirmation of eligibility, including a Rapid Antigen Test (RAT) for 
SARS-CoV-2, each participant will be inoculated intravenously on Day 0 with approximately 2,800 viable 
P. falciparum-infected erythrocytes. Participants will be followed up daily via phone call or text message 
on Days 1 to 3 post-inoculation to solicit any adverse events. 
 
Participants will attend the clinical unit once on Days 4, 5, 6 and 7 for clinical evaluation and blood 
sampling to monitor the progression of parasitaemia, using quantitative polymerase chain reaction (qPCR) 
targeting the gene encoding 18S rRNA (referred to hereafter as malaria 18S qPCR). Participants will have 
a nasopharyngeal aspirate (NPA) for SARS-CoV-2 on Day 6 am. Participants will also have blood collected 
on Day 7 am to monitor haematology and biochemistry.  
 
Participants will be admitted to the clinical trial unit on Day 8 for dosing with the investigational medicinal 
product (IMP; pyronaridine) when parasitaemia for the majority of participants is expected to be above 
5,000 parasites/mL. Participants will need to return a negative (RAT) for SARS-CoV-2 prior to admission. 
Pyronaridine will be administered as a single oral dose. Different doses of pyronaridine will be 
administered across and within cohorts in order to effectively characterise the PK/PD relationship. Each 
cohort will comprise up to three dose groups, with participants randomised to a dose group on the day of 
dosing. The highest dose of pyronaridine administered will be no more than 720 mg; the lowest dose 
administered will be no less than 180 mg. The proposed dosing regimen for the Cohort 1, assuming six 
participants are enrolled, is as below. If less than six participants are enrolled the Safety Data Review Team 
(SDRT) will meet between Day 0 and Day 8, to decide on the dose/s to be administered. 
 
Dose group 
 
1A (n=2) 
1B (n=2) 
1C (n=2) 
Pyronaridine 
360 mg  
540 mg 
720 mg  
Each subsequent cohort will be composed of up to 3 dose groups. If more than one dose is tested in a 
given cohort, subjects will be randomised after inoculation day but prior to Day 8. The dose/s 
administered to Cohort 2 and Cohort 3 will be selected based on PK/PD and safety data from the preceding 
cohort/s. The SDRT will review all available safety and tolerability data and PK/PD analysis outcomes from 
the previous cohort/s, prior to inoculation of the next cohort. The study will conclude when sufficient data 
have been obtained to define the PK/PD parameters for pyronaridine (the primary endpoint).    


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20 
 
A maximum sample size of 18 participants administered pyronaridine is expected to be sufficient to 
achieve the primary endpoint. 
Participants will be confined in the clinical unit for at least 96 h (Days 8 - 12) to monitor the safety and 
tolerability of pyronaridine dosing and to ensure adequate clinical response against P. falciparum. During 
confinement, regular safety assessments will be performed and blood will be collected to monitor parasite 
clearance and pyronaridine concentration. If participants are clinically well at the end of the confinement 
period, they will be discharged and monitored on an outpatient basis up to Day 50±2.  
Participants 
will 
receive 
compulsory 
antimalarial 
rescue 
treatment 
with 
Riamet® 
(artemether/lumefantrine) on Day 47±2, or earlier in the following cases: 
 
Parasitaemia is not reduced by approximately 10-fold at the end of the 96 hour confinement 
period (Day 12) when compared with peak parasitaemia. 
 
Parasite regrowth occurs after an initial reduction in parasitaemia, with regrowth defined as 
≥5000 parasites/mL and a two-fold parasitaemia increase within 48 hours.  
 
Volunteer discontinuation/withdrawal from the study. 
 
Investigator’s discretion in the interest of participant safety. 
Parasite lifecycle stages (including gametocytes) will be monitored at select time points during the study 
by collecting blood samples and using quantitative reverse-transcriptase polymerase chain reaction (qRT-
PCR) targeting parasite lifecycle stage-specific mRNA transcripts. 
If gametocytes are present as determined by qRT-PCR any time point after the confinement period, a 
single dose of Primacin® will be administered at the Investigator’s discretion.  
If an allergy or contraindication to Riamet® develops, Malarone® (atovaquone/proguanil) will be 
administered. If in the rare event participants are unable to complete oral antimalarial rescue treatment, 
they will be admitted to hospital to receive intravenous artesunate.  
4.2 
SCIENTIFIC RATIONALE FOR STUDY DESIGN 
 
The study design of this trial using the IBSM model has been used previously to safely and effectively 
determine the PK/PD relationship of several antimalarial agents [6-9]. The P. falciparum 3D7 challenge 
agent used to initiate blood-stage malaria infection in this study has been well characterised, with 
consistent and reproducible parasite growth occurring following intravenous inoculation of healthy 
participants in previous studies. Experience from these studies indicates that parasitaemia will be above 
5,000 parasites/mL on Day 8 following inoculation, but clinical symptoms of malaria will either be absent 
or only mild in severity.  
 
The duration of confinement within the clinic following dosing with pyronaridine on Day 8 (96 hours) is 
considered appropriate to ensure close monitoring of participants for safety purposes and to enable 
frequent blood sampling for parasitaemia and drug concentration measurements. Blood sampling time 
points have been selected by taking into account data from previous clinical PK studies, as well as 


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21 
 
preclinical data on the antimalarial activity of pyronaridine. These time points may be adjusted during the 
study, by means of a protocol amendment, based on the emerging data in order to optimally determine 
the exposure/response relationship of pyronaridine.  
 
The total follow-up period following pyronaridine dosing (42±2 days) is equal to three times the 
elimination half-life of pyronaridine (approximately 14 days). This duration was selected to allow sufficient 
time for pyronaridine blood concentrations to fall to a level amenable to parasite regrowth, which is 
required to calculate the PK/PD parameters. Further, this duration will ensure that participants have a 
negligible blood concentration of the IMP at the completion of the study. 
 
Compulsory treatment with the registered antimalarial Riamet® (artemether/lumefantrine) will occur for 
all participants to ensure all parasites are cleared at the completion of the study. P. falciparum 3D7 
parasites are known to be sensitive to artemether/lumefantrine, and Riamet® has been effective in 
clearing parasitaemia in previous IBSM studies using this challenge agent. A single dose of Primacin® will 
be used to ensure clearance of gametocytes (the life cycle stage responsible for transmission to 
mosquitoes) if these are present at any time point after the confinement period at the Investigator’s 
discretion. An alternate registered antimalarial treatment Malarone® (atovaquone/proguanil) will be used 
in the event of contraindication to Riamet® Malarone has also previously been effective in clearing P. 
falciparum 3D7 parasitaemia [22]. If in the rare event participants are unable to complete oral antimalarial 
rescue treatment, they will admitted to hospital to receive intravenous artesunate, the recommended 
parenteral treatment of malaria in Australia.  
4.3 
JUSTIFICATION FOR DOSE 
 
Pyronaridine will be administered as a single oral dose on Day 8, with different doses to be tested across 
and within cohorts in order to effectively characterise the PK/PD relationship. Each cohort will comprise 
up to three dose groups, with participants randomised to a dose group on the day of dosing. For each 
cohort, the SDRT will meet between Day 0 and Day 8, to decide on the dose/s to be administered. Dose 
selection will be decided based on number of participants inoculated with the malaria challenge agent, 
and when applicable available data from previous cohorts. 
 
The highest dose of pyronaridine administered will be no more than 720 mg. This dose is equal to the 
maximum single daily dose of pyronaridine administered to adult patients ≥65 kg treated with Pyramax® 
(pyronaridine/artesunate combination) in accordance with manufacturer’s product information for the 
treatment of acute uncomplicated malaria. Pyramax® is administered as a multiple dose treatment course 
(daily dosing with 720 mg pyronaridine/ 240 mg artesunate over three days for adults ≥65 kg), thus the 
total exposure to pyronaridine following administration of a single dose of 720 mg in the current study is 
expected to be lower than occurs during a course of Pyramax® treatment. A dose of 720 mg may be 
required to determine the maximum killing rate (Emax) of pyronaridine in the IBSM model.   
 


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22 
 
The lowest dose of pyronaridine administered in this study will be no less than 180 mg. Modelling studies 
using clinical PK data and preclinical PD data (in vitro and murine studies) have indicated that a dose as 
low as 180 mg may be required for parasite regrowth to occur following initial parasite clearance. Parasite 
regrowth is required in this study to calculate all PK/PD parameters (primary endpoint).  
4.4 
END OF STUDY DEFINITION 
 
A participant is considered to have completed the trial if they have completed all phases of the trial 
including the last visit or the last scheduled procedure shown in the Schedule of Activities (SoA), Section 
1.4. For the purposes of clarity, a participant may still be considered to have completed the trial if they 
have not completed all scheduled procedures. The end of the trial globally is defined as the time at which 
all participants have completed the trial


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23 
5 
STUDY POPULATION 
 
5.1 
INCLUSION CRITERIA 
Individuals must fulfil all of the following criteria to be eligible for inclusion in this trial: 
Demography 
1. 
Male or female (non-pregnant, non-lactating) aged 18 to 55 years inclusive who will be 
contactable and available for the duration of the trial and up to two weeks following the EOS visit. 
2. 
Total body weight greater than or equal to 50 kg, and a body mass index (BMI) within the range 
of 18 to 32 kg/m2 (inclusive). BMI is an estimate of body weight adjusted for height. It is calculated 
by dividing the weight in kilograms by the square of the height in metres. 
Health status 
3. 
Certified as healthy by a comprehensive clinical assessment (detailed medical history and full 
physical examination). 
4. 
Fully vaccinated (meaning first, second and booster dose) against COVID-19 within 14 days of 
planned inoculation date. 
5. 
Vital signs at screening (measured after 5 min in the supine position): 
 
Systolic blood pressure (SBP) - 90–140 mmHg, 
 
Diastolic blood pressure (DBP) - 40–90 mmHg, 
 
Heart rate (HR) 40–100 bpm. 
6. 
At Screening and pre-inoculation with the malaria challenge agent: normal standard mean of 
triplicate 12-lead electrocardiogram (ECG) parameters after 5 minutes resting in supine position 
in the following ranges:  
a. QTcF ≤450 msec (male participants); QTcF ≤470 msec (female participants);  
b. QRS  50–120 msec 
c. PR interval ≤ 210 msec for both males and females, and  
d. Normal ECG tracing unless the PI or delegate considers an ECG tracing abnormality to be 
not clinically relevant.  
7. 
Women of childbearing potential (WOCBP) who anticipate being sexually active with a male 
during the trial must agree to the use of a highly effective method of birth control (see below) 
combined with a barrier contraceptive from the screening visit until 100 days after the last dose 
of pyronaridine (covering a full menstrual cycle of 30 days starting after 5 half-lives of last dose 
pyronaridine) and have a negative result on urine pregnancy test performed before inoculation 
with the malaria challenge agent.  
Note:  


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24 
a. Highly effective birth control methods include: combined (oestrogen and progestogen 
containing) oral/intravaginal/transdermal/implantable 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 or same sex relationship. 
b. Female participants who are abstinent (from penile-vaginal intercourse) must agree to start 
a double method if they start a sexual relationship with a male during the study. Female 
participants must not be planning in vitro fertilisation within the required contraception 
period.  
8. 
Women of non-childbearing potential (WONCBP) are defined as: 
a. Natural (spontaneous) post-menopausal defined as being amenorrhoeic for at least 12 months 
without an alternative medical cause with a screening follicle stimulating hormone level (FSH) >25 
IU/L (or at the local laboratory levels for post-menopause) 
b. Premenopausal with irreversible surgical sterilization by hysterectomy and/or bilateral 
oophorectomy or salpingectomy at least 6 months before screening (as determined by participant 
medical history) 
9. 
Males who have, or may have, female sexual partners of child bearing potential during the course 
of the study must agree to use a double method of contraception including condom plus 
diaphragm, 
or 
condom 
plus 
intrauterine 
device, 
or 
condom 
plus 
stable 
oral/transdermal/injectable/implantable hormonal contraceptive by the female partner, from the 
time of informed consent through to 70 days (covering a spermatogenesis cycle) after 
pyronaridine administration. Abstinent males must agree to start a double method if they begin 
sexual relationship with a female during the study and up to 70 days after the last dose of 
pyronaridine. Males with female partners of child-bearing potential that are surgically sterile, or 
males who have undergone sterilisation and have had testing to confirm the success of the 
sterilisation, may also be included and will not be required to use above described methods of 
contraception. 
Regulations 
10. 
Completion of the written informed consent process prior to undertaking any trial-related 
procedure. 
11. 
Must be willing and able to communicate and participate in the whole trial. 
12. 
Agreement to adhere to Lifestyle Considerations (see Section 5.3) throughout trial duration 
13. 
Agreement 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. 
 
 


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25 
5.2 
EXCLUSION CRITERIA 
 
1. Individual who lives alone, OR, does not satisfy the following criteria: Participants who live alone 
may be included on a case-by-case basis, following discussion with the Principal 
Investigator.  Participants who live alone must identify and provide contact details of a support 
person who is aware of the participant’s participation in the study and is available to provide 
assistance if required (for example with contacting the participant in the event that study staff 
are unable to, or with transporting the participant to and from the study site if required). 
2. Known hypersensitivity to pyronaridine, artesunate or any of its derivatives, artemether, 
lumefantrine or other artemisinin derivatives, proguanil/atovaquone, primaquine, or 4-
aminoquinolines. 
3. Haematology, biochemistry or urinalysis results at screening or at the eligibility visit that are 
outside of Sponsor-approved clinically acceptable laboratory ranges (appendix) or are considered 
clinically significant by the PI or their delegate.  
 
4. Participation in any investigational product trial within the 12 weeks preceding pyronaridine 
administration. 
5. Symptomatic postural hypotension at screening (confirmed on two consecutive readings), 
irrespective of the decrease in blood pressure, or asymptomatic postural hypotension defined as 
a decrease in systolic blood pressure ≥20 mmHg within 2–3 min when changing from supine to 
standing position. 
6. Any history of anaphylaxis or other severe allergic reactions including face, mouth, or throat 
swelling or any difficulty breathing, or other food or drug allergy that the Investigator considers 
may impact on participant safety. Participants with seasonal allergies/hay fever or allergy to 
animals or house dust mite that are untreated and asymptomatic at the time of dosing can be 
enrolled in the trial.  
7. History of convulsion (including drug or vaccine-induced episodes). A medical history of a single 
febrile convulsion during childhood (< 5 years) is not an exclusion criterion. 
8. Presence of current or suspected serious chronic diseases such as cardiac or autoimmune disease, 
diabetes, progressive neurological disease, severe malnutrition, hepatic or renal disease. Acute or 
progressive hepatic or renal disease, porphyria, psoriasis, rheumatoid arthritis, asthma (excluding 
childhood asthma, or mild asthma with preventative asthma medication required less than 
monthly), or epilepsy. 
9. History of malignancy of any organ system (other than localised basal cell carcinoma of the skin 
or in situ cervical cancer), treated or untreated, within five years of screening, regardless of 
whether there is evidence of local recurrence or metastases. 
10. Individuals with history of schizophrenia, bipolar disorder psychoses, disorders requiring lithium, 
attempted or planned suicide, or any other severe (disabling) chronic psychiatric diagnosis 
including generalised anxiety disorder. 
11. Individuals who have been hospitalised within five years prior to enrolment for either a psychiatric 
illness or due to danger to self or others. 


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26 
12. History of an episode of mild/moderate depression lasting more than 6 months that required 
pharmacological therapy and/or psychotherapy within the last 5 years; or any episode of major 
depression. 
The Beck Depression Inventory-II (BDI-II) will be used as a validated 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 BDI-II 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. These participants will be referred to a general 
practitioner or medical specialist as appropriate. Participants with a BDI-II score of 17 to 19 may be 
enrolled at the discretion of an 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 participant or to the execution of the trial and interpretation of the data gathered. 
13. History of recurrent headache (e.g. tension-type, cluster, or migraine) with a frequency of ≥2 
episodes per month on average and severe enough to require medical therapy, during the 2 years 
preceding screening. 
14. Presence of clinically significant infectious disease or fever (e.g., sublingual temperature ≥38.5°C) 
within the five days prior to inoculation.  
15. 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 8.  
16. History or presence of alcohol abuse (alcohol consumption more than 40 g/4 units/4 standard 
drinks per day), or drug habituation, or any prior intravenous usage of an illicit substance. 
17. Any individual who currently (within 14 days prior to inoculation) smokes >5 cigarettes/day. 
18. Breastfeeding or lactating; positive serum pregnancy test at screening, positive urine pregnancy 
test upon admission or at other time points as specified by schedule of activities tables.  
19. Any COVID-19 vaccine within 14 days of inoculation, any other vaccination within 28 days of IMP 
intake, and any vaccination planned up to the final follow-up visit. 
20. Any corticosteroids, anti-inflammatory drugs (excluding commonly used over-the-counter anti-
inflammatory drugs such as ibuprofen, acetylsalicylic acid, diclofenac), immunomodulators or 
anticoagulants within the past three months. Any individual currently receiving or having 
previously 
received 
immunosuppressive 
therapy 
(including 
systemic 
steroids, 
adrenocorticotrophic hormone or inhaled steroids) at a dose or duration potentially associated 
with hypothalamic-pituitary-adrenal axis suppression within the past year. 
21. Use of prescription drugs (excluding contraceptives) or non-prescription drugs or herbal 
supplements (such as St John’s Wort), within 14 days or five half-lives (whichever is longer) prior 
to inoculation. Limited use of other non-prescription medications or dietary supplements, not 
believed to affect participant safety or the overall results of the trial, may be permitted on a case-
by-case basis following approval by the Sponsor in consultation with the PI. Participants are 
requested to refrain from taking non-approved concomitant medications from recruitment until 
the conclusion of the trial. 
22. Cardiac/QT risk: 
 
Family history of sudden death or of congenital prolongation of the QTc interval or known 
congenital prolongation of the QTc interval or any clinical condition known to prolong the QTc 
interval. 


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27 
 
History of symptomatic cardiac arrhythmias or with clinically relevant bradycardia. 
23. Any history of malaria or participation in a previous malaria challenge trial or malaria vaccine trial. 
24. Must not have had malaria exposure that is considered by the PI or their delegate to be significant. 
This includes but is not limited to: history of having travelled to or lived (>2 weeks) in a malaria-
endemic region during the past 12 months or planned travel to a malaria-endemic region during the 
course of the trial; history of having lived for >1 year in a malaria-endemic region in the past 10 
years; history of having ever lived in a malaria-endemic region for more than 10 years inclusive. For 
endemic regions see https://malariaatlas.org/explorer/#/, Bali is not considered a malaria-endemic 
region. 
25. Has evidence of increased cardiovascular disease risk (defined as >10%, 5-year risk for those 
greater than 35 years of age, as determined by the Australian Absolute Cardiovascular Disease 
Risk Calculator [http://www.cvdcheck.org.au/]). Risk factors include sex, age, systolic blood 
pressure (mm/Hg), smoking status, total and HDL cholesterol (mmol/L), and reported diabetes 
status. 
26. History of splenectomy. 
27. Individual unwilling to defer blood donations to the Blood Service for at least twelve months after 
the EOS visit. 
28. Individual who has ever received a blood transfusion. 
29. Any recent (<6 weeks) therapy with an antibiotic or drug with potential antimalarial activity (e.g. 
chloroquine, piperaquine phosphate, benzodiazepine, flunarizine, fluoxetine, tetracycline, 
azithromycin, clindamycin, doxycycline etc.). 
General conditions 
30. Any individual who, in the judgement of an Investigator, is likely to be non-compliant during the 
trial, or is unable to cooperate because of a language problem or poor mental development. 
31. Any individual in the exclusion period of a previous trial according to applicable regulations. 
32. Any individual who is an Investigator, research assistant, pharmacist, trial coordinator, or other 
staff thereof, directly involved in conducting the trial. 
33. Any individual without good peripheral venous access. 
 
Biological status 
34. Positive result on any of the following tests: hepatitis B surface antigen (HBs Ag), anti-hepatitis B 
core antibodies (anti-HBc Ab), anti-hepatitis C virus (anti-HCV) antibodies, anti-human 
immunodeficiency virus 1 and 2 antibodies (anti-HIV1 and anti-HIV2 Ab). 
35. Positive urine drug test. Any drug listed in the urine drug screen unless there is an explanation 
acceptable to an Investigator (e.g., the participant has stated in advance that they consumed a 
prescription or over-the-counter product that contained the detected drug) and the participant 
has a negative urine drug screen on retest by the pathology laboratory.  
36. Severe G6PD deficiency. 
37. Positive alcohol breath test. 
38. Positive for SARS-CoV-2 by PCR or RAT.  


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28 
39. Positive for red cell antibodies 
5.3 
LIFESTYLE CONSIDERATIONS 
While participating in this trial, participants are asked to: 
 
Refrain from alcohol consumption of more than 20 g/2 units/2 standard drinks per day from 24 hours 
before inoculation until the EOS. 
 
Abstain from any illicit drug habituation until the EOS. 
 
Abstain from any alcohol and tobacco use for the duration of clinical trial unit confinement. 
 
Refrain from tobacco use of more than five cigarettes or equivalent per day until the EOS.  
 
Refrain from excessive consumption of beverages or food containing xanthine bases including Red 
Bull, chocolate, coffee etc. (more than 400 mg caffeine per day, equivalent to more than 4 cups of 
coffee per day). 
 
Refrain from consumption of Seville oranges, and grapefruit or grapefruit juice from 7 days prior to 
inoculation until EOS.  
 
Refrain from consumption of quinine containing foods/beverages such as tonic water and lemon 
bitter from inoculation until the EOS. 
 
Abstain from strenuous exercise for 24 h before each blood collection for clinical laboratory tests. 
Participants may participate in light recreational activities during studies. 
5.4 
SCREEN FAILURES 
Healthy candidate participants who do not fulfil all the inclusion criteria, and/or fulfil any of the exclusion 
criteria should not be enrolled, however 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. A positive urine drug screen should only be re-tested if there is a strong rationale for doing so 
(for example, false positive). The result of the re-test must be considered for participant eligibility and 
must be available prior to inoculation or pyronaridine administration. In case of doubt, the PI is to confer 
with the Medical Monitor for agreement.   
If a participant does not meet all eligibility 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 one occasion only. 
Participants who are rescreened should be assigned the same screening number as for the initial 
screening. 
A minimal set of screen failure information is required to ensure transparent reporting of screen failure 
participants, to meet the Consolidated Standards of Reporting Trials (CONSORT) publishing requirements 
and to respond to queries from regulatory authorities. Minimal information includes demography, screen 
failure details, eligibility criteria, and any serious adverse event (SAE). Screen failure information will not 
be entered into the eCRF. Participants who fail screening due to an underlying medical condition 
previously unknown to them will be reimbursed for their time, and provided with the appropriate referrals 
for guidance and counselling for their condition. 


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29 
5.5 
STRATEGIES FOR RECRUITMENT AND RETENTION 
It is anticipated that up to 18 healthy, adult, malaria- naïve participants will need to be dosed with 
pyronaridine in this trial. It is anticipated that a total of 6 reserve participants will be required to ensure 
adequate participant numbers.  
Participants will be recruited by general or trial specific advertising via print, radio, social media, or poster 
media, as approved by the HREC(s). No restrictions will apply for ethnic or racial categories.  
Participants who withdraw or are withdrawn from the trial will be compensated on a fractional basis for 
their involvement unless they are withdrawn as a consequence of their misconduct.  
Reserve participants who do not participate in the trial will be compensated for the inconvenience 
associated with their attendance for screening and attendance on Day 0. 
The monetary value of the compensation for each trial part is documented in the respective Participant 
Information Sheet.  
6 
STUDY INTERVENTIONS 
 
6.1 
STUDY INTERVENTIONS ADMINISTRATION 
 
The following sections describe the study interventions. A pharmacy manual with detailed information 
will be finalised and approved prior to study start. 
 
6.1.1 STUDY INTERVENTIONS DESCRIPTION 
 
IMP 
Pyronaridine is a benzonaphthyridine derivative first synthesized in China in 1970. Pyronaridine has been 
used in China for the treatment of malaria as a single agent for more 50 years. The antimalarial activity of 
pyronaridine is thought to occur primarily due to inhibition of hemozoin formation, which results in 
accumulation of toxic hematin in the parasite food vacuole. 
 
It was developed as an oral fixed-dose combination therapy with artesunate (an artemisinin derivative) 
for the treatment of acute uncomplicated P. falciparum malaria and blood stage P. vivax (Pyramax®, co-
developed by Medicines for Malaria Venture, Geneva, Switzerland and Shin Poong Pharmaceutical Co., 
Ltd, Seoul, South Korea). Pyramax® received a positive opinion by European Medicines Agency (EMA) 
under the article 58 process [evaluation of medicinal products intended exclusively for markets outside 
the European Community] in 2012 and 2015. Pyramax® was also added to the World Health 
Organization’s (WHO) Model List of Essential Medicines (EML) and Model List of Essential Medicines for 
Children (EMLc) in 2017. Its procurement and use at a country level is further supported by a WHO 
Information Note of October 2019. Pyramax® is currently approved for use in 29 countries, although it has 
not yet been registered for use in Australia. For the current study, pyronaridine tablets for oral 


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30 
administration will be manufactured by Shin Poong Pharmaceutical Co., Ltd, licence holder and 
manufacturer of Pyramax.  
 
Challenge agent 
A P. falciparum 3D7 master cell bank (MCB) was produced from a participant with type O Rh(D) negative 
blood who was infected with the parasite by mosquito bite. Blood was collected and aliquoted into 
cryovials and stored in liquid nitrogen under controlled conditions. Refer to the P. falciparum 3D7 IB for 
more details. A cryovial will be retrieved from storage, thawed, and used to aseptically prepare the 
intravenous inoculum at Q-Gen Cell Therapeutics (QIMR Berghofer). 
Rescue Antimalarial medications 
Riamet® (Novartis Pharmaceuticals Australia Pty Ltd) is an oral fixed dose combination therapy containing 
artemether and lumefantrine. Riamet® is registered in Australia for the treatment of acute uncomplicated 
malaria.  
Primacin® (Boucher & Muir Pty Ltd) contains the active ingredient primaquine phosphate. Primaquine 
phosphate is an 8-aminoquinoline effective against the sexual forms (gametocytes) of Plasmodium species 
which are responsible for transmission to mosquitoes. Primacin® is registered for use in Australia.  
Malarone® (GlaxoSmithKline Australia Pty Ltd) is an oral fixed dose combination therapy containing 
atovaquone and proguanil hydrochloride. Malarone® is registered in Australia for the treatment of acute 
uncomplicated malaria.  
Artesunate (powder and diluent for reconstitution, imported by Link Pharmaceuticals) is an artemisinin 
derivative; the intravenous formulation is typically used to treat severe malaria. Although parenteral 
artesunate does not currently have marketing approval in Australia, the Therapeutic Goods 
Administration (TGA) allows importation of artesunate for Category A use in patients with severe malaria 
under the Special Access Scheme (SAS). 
6.1.2 DOSING AND ADMINISTRATION 
 
IMP 
Pyronaridine tablets will be administered after an overnight fast of ≥ 8 hours as a single oral dose with 
240 mL of water on Day 8. Pyronaridine will not be administered if any of the criteria described in 7.1.1 
has been met. In this situation participants will be administered antimalarial rescue treatment. Different 
doses of pyronaridine will be tested across and within cohorts, with the doses to be determined by the 
SDRT based on the emerging data during the study. The maximum dose to be tested in the study will be 
no more than 720 mg (4 tablets). The minimum dose to be tested in the study will be no less than 180 mg 
(1 tablet).   
 
Challenge Agent 
The malaria challenge agent, containing an estimated ~2,800 viable P. falciparum 3D7 parasite-infected 
erythrocytes in a volume of 2 mL, will be administrated intravenously on Day 0. The actual number of 


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31 
parasites inoculated will take into account the loss of viability resulting from cryopreservation, storage 
and thawing. Participants will undergo intravenous cannulation with a 20 or 22 gauge cannula. Placement 
and patency will be checked by flushing the vein with 5-10 mL of clinical grade saline. The inoculum will 
be injected, and the cannula again flushed with 5-10 mL of clinical grade saline. The cannula will then be 
removed, and haemostasis ensured by use of an appropriate dressing. An extra syringe will be prepared 
to quantify the parasite count of the challenge agent by malaria 18S qPCR. 
Rescue Antimalarial medications 
Compulsory definitive antimalarial treatment with Riamet® for all participants will be initiated on Day 
47±2. Treatment may be initiated earlier in accordance with the criteria described in Section 4.1. Riamet® 
tablets will be administered as six oral doses of four tablets (total course of 24 tablets) given over a period 
of 60 hours (total dose of 480 mg artemether and 2.88 g lumefantrine). Participants will be reminded of 
the potential side effects of Riamet® and will be given the CMI for Riamet®. 
Participants may be treated with Primacin® if gametocytaemia is suspected from parasite lifecycle stage 
qRT-PCR at the Investigator’s discretion, to ensure clearance of gametocytes. If needed, participants will 
be administered a single oral dose of six Primacin® tablets (total dose 45 mg primaquine) with food, unless 
G6PD deficient in which case a low dose of 15 mg (two tablets) will be administered. 
Participants will be reminded of the potential side effects of Primacin® and will be given the CMI for 
Primacin®. 
If an allergy or contraindication to Riamet® develops, Malarone® may be administered at the 
Investigator’s discretion. A treatment course consists of four tablets administered once daily for three 
days (total dose of 3 g atovaquone and 1.2 g proguanil hydrochloride). Participants will be reminded of 
the potential side effects of Malarone® and will be given the CMI for Malarone®. 
Intravenous artesunate may be administered if the participant vomits or cannot tolerate oral drugs. In 
this rare event, the participant will be admitted to hospital for treatment. Artesunate will be administered 
as 2.4 mg/kg IV bolus on admission, with repeat dosing at 12 hours and 24 hours, then once daily until 
oral therapy is possible. 
 
 


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32 
6.2 
PREPARATION/HANDLING/STORAGE/ACCOUNTABILITY 
6.2.1 ACQUISITION AND ACCOUNTABILITY 
 
The clinical trial unit pharmacist or delegate, as nominated by the PI, is responsible for maintaining 
accurate study intervention accountability records throughout the study. Study interventions include the 
malaria challenge agent, the IMP (pyronaridine), and the antimalarial rescue treatments. Dispensing, 
accountability and documentation will be in accordance with the clinical unit standard procedures.  
All products will be inventoried upon receipt by the clinical trial unit pharmacist. The condition of the 
products at the time of receipt by the pharmacist will be documented, as will the time restrictions of use 
for the syringes containing the challenge agent. The lot numbers and expiry dates of all study interventions 
will be documented. The clinical trial unit pharmacist or delegate will ensure that the received products 
are the specified formulation.  
The storage, handling and the disposal of the challenge agents will be in accordance with approved 
procedures. All dosages prescribed and dispensed to the participants and all dose changes during the 
study must be recorded in the eCRFs and accountability logs. All drug supplies are to be used only in 
accordance with this protocol, and not for any other purpose. All used medications will be fully 
documented. Used and unused drug containers must be destroyed at the unit once drug accountability is 
final and has been checked by the Sponsor or its delegate, and written permission for destruction has 
been obtained from the Sponsor. 
Study products and study accountability logs will be available to the Sponsor or their representative as 
part of the study monitoring procedures. Upon completion of the study, copies of all study drug 
management records will be provided to the Sponsor. Original records will be maintained at the clinical 
unit with the rest of the study records. 
IMP 
Pyronaridine tablets will be manufactured by Shin Poong Pharmaceutical Co., Ltd, Seoul, South Korea 
and acquired by the clinical unit.  
 
Challenge agent 
Aliquots of the P. falciparum 3D7 MCB are stored in liquid nitrogen under controlled conditions. On the 
day of inoculation, cryovials will be retrieved from storage and used to aseptically prepare the challenge 
agent at Q-Gen Cell Therapeutics (QIMR Berghofer). The clinical trial unit pharmacist or designee will be 
responsible for maintaining the accurate Plasmodium falciparum 3D7 Challenge Agent Accountability Log 
as per clinical trial unit standard operating procedures (SOPs). 
Rescue Antimalarial medications 
Riamet® (distributed by Novartis Pharmaceuticals Pty Ltd), Primacin® (distributed by Boucher & Muir Pty 
Limited), and Malarone® (distributed by GlaxoSmithKline Australia Pty Ltd) will be acquired by the clinical 


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33 
trial unit. Artesunate for intravenous administration is available within the Central Pharmacy of 
Queensland Health.  
6.2.2 FORMULATION, APPEARANCE, PACKAGING, AND LABELING 
 
The contents of the dispensing labels for the IMP, the malaria challenge agent, and the antimalarial rescue 
treatments will be in accordance with all applicable regulatory requirements. 
IMP 
Pyronaridine tablets, each containing 180 mg pyronaridine tetraphosphate, are orange, round, film-
coated scored tablets. Excipients include: microcrystalline cellulose, crospovidone, mannitol, magnesium 
stearate, talc, Hypromellose 2910, macrogol 6000 and Opadry OY-S-23068. Tablets are packaged in 
aluminium/PVC/aluminium-oPA foil blisters containing 9 tablets per blister. The blisters are packed into 
cartons. 
Challenge agent 
The malaria challenge agent will contain parasitised and unparasitised erythrocytes, resuspended in 0.9% 
Sodium Chloride Intravenous Infusion, in a total volume of 2 mL in syringes. The syringes will be double 
contained following preparation and labelled in accordance with GCP guidelines. Labelling is in accordance 
with PIC/S Annex 13. 
Rescue Antimalarial medications 
Riamet® tablets (each containing 20 mg artemether/120 mg lumefantrine) are yellow, round, flat tablets 
marked with N/C and a score line on one side and CG on the other side. Each carton contains 24 tablets. 
Primacin® tablets (each containing 13.2 mg primaquine phosphate equivalent to 7.5 mg primaquine) are 
round, flat, orange uncoated tablets available in bottles of 28 or 56 tablets. 
Malarone® tablets are round, pink and film-coated, and are engraved with “GX CM3”. Malarone® tablets 
are supplied in blister packs of 12 or 24 tablets. 
Artesunate powder for reconstitution is supplied in vials containing 60 mg artesunate. Each vial is 
reconstituted with an ampoule (1 mL) of sodium bicarbonate 5%. Sodium chloride 0.9 % (5 mL) is then 
added to reconstituted artesunate vial to create a 10 mg/mL solution (total volume 10 mL).  
6.2.3 PRODUCT STORAGE AND STABILITY 
 
All drugs will be held in appropriate locked storage conditions at the clinical trial unit until required. 
 
IMP 
Pyronaridine tablets are to be stored below 25°C in the original package. The shelf life is 18 months.  
 
Challenge agent 


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34 
The P. falciparum 3D7 challenge agent is prepared at Q-Gen on inoculation day (Day 0). The time between 
preparation of the challenge agent and administration to the participant will be a maximum of 4 hours. 
The syringes containing the challenge agent will be stored in a temperature monitored validated transport 
container at 2-15°C during transportation from Q-Gen to the clinical trial unit and will be immediately 
transferred to a temperature monitored onsite refrigerator (2-15°C). The clinical trial unit pharmacist will 
document receipt conditions and time restrictions of use. The challenge agent will then be dispensed to 
participants as per written prescription. Transporter logs should be provided to the Sponsor. 
 
Rescue Antimalarial medications 
Riamet® tablets are to be stored below 30°C and protected from moisture. Primacin® tablets are to be 
stored below 25°C. Malarone® tablets are to be stored below 30°C. The reconstituted artesunate for 
intravenous administration should be prepared freshly for each administration and should not be stored.  
 
6.2.4 PREPARATION 
 
IMP 
No preparation is required for the pyronaridine tablets for oral administration.  
 
Challenge agent 
The challenge agent will be prepared aseptically at Q-Gen from frozen cryovials of the P. falciparum 3D7 
MCB by nominated QIMR Berghofer staff members. The erythrocytes will be thawed, washed, 
resuspended in saline, diluted in a final volume of 2 mL of clinical grade saline, and dispensed into syringes. 
Any remaining unused erythrocytes will be discarded as per approved SOPs. 
 
Rescue Antimalarial medications 
Riamet®, Primacin® and Malarone® are available as tablets and no preparation is required. Artesunate for 
intravenous administration will be prepared according to manufacturer’s instructions within the hospital 
setting immediately prior to administration.  
6.3 
MEASURES TO MINIMIZE BIAS: RANDOMISATION  
 
All participants will receive an identification number (screening number) as soon as they have signed the 
informed consent form. Participants who meet the eligibility criteria and are enrolled in the study will be 
assigned an enrolment number on Day 0 immediately prior to administration of the malaria challenge 
agent. Participants will be randomised within each cohort to a dose group on Day 8, immediately prior to 
administration of the IMP. This is an open label study and therefore no blinding will be performed. 
6.4 
STUDY INTERVENTIONS COMPLIANCE 
 
The investigator will administer the malaria challenge agent intravenously at the clinical trial unit on Day 
0.  
 


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35 
The IMP (single oral dose of pyronaridine) will be administered at the clinical trial unit on Day 8 under 
direct observation by staff.  
 
The first dose of Riamet® will be administered at the clinical trial unit under direct observation by staff. 
The subsequent five doses may be taken at home. Participants will receive a phone call or text message 
from the clinical trial unit staff to ensure compliance.  
 
If Primacin® is required at any time point after the confinement period, it will be administered at the 
clinical trial unit under direct observation by staff. 
 
If Malarone® is required, the first dose will be administered at the clinical trial unit under direct 
observation by staff. The subsequent two doses may be taken at home. 
 
Participants will receive a phone call or text message from the clinical trial unit staff to ensure compliance. 
If a participant requires intravenous artesunate, the participant will be admitted to hospital for treatment. 
6.5 
CONCOMITANT THERAPY 
 
Concomitant medications, treatments and procedures are those occurring from the time of 
administration of the malaria challenge agent (Day 0), until the final study visit. Those occurring prior to 
Day 0 are classified as prior medications, treatments and procedures. Medications taken within 28 days 
before Day 0 will be recorded as prior medication.  
On Day 0, participants will be questioned in relation to relevant aspects of compliance with the study 
protocol, including drug intake since their screening visit. Details of all drugs taken (prescription and over-
the-counter, systemic and topical administration) will be recorded at this time and appropriate action 
taken.  
From Day 0 until the EOS visit, no medications will be permitted to be taken without the approval of the 
PI (with the exception of contraceptives and ibuprofen). Ibuprofen (preferred) may be administered at 
doses of up to 1.2 g/24 hours, or paracetamol up to 4 g/24 hours with Investigator approval, for treatment 
of headache or other pain if required. Ibuprofen is the preferred treatment for headache or pain. To 
minimise the risk of liver enzyme elevation, paracetamol is to be avoided if possible; however paracetamol 
may be required by some participants and as such is not a prohibited substance. All other medications 
will be assessed by the PI on a case by case basis with regard to the participant’s wellbeing and potential 
for the medication to interfere with study interventions.   
Any medication taken during the study for treatment of a medical condition or AE is to be recorded in the 
concomitant medication pages in the eCRF. The exact dose, route and timing of each dose should be 
recorded. 
 


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36 
6.5.1 RESCUE MEDICINE 
   
Antimalarial rescue treatments used in this study are Riamet®, Primacin® (if required), Malarone® (if 
required), and IV artesunate (if required). Details are provided in Sections 6.1 and 6.2. 
 
7 
STUDY INTERVENTIONS DISCONTINUATION AND PARTICIPANT 
DISCONTINUATION/WITHDRAWAL 
7.1 
DISCONTINUATION OF STUDY INTERVENTIONS 
The Sponsor, SDRT, PI, approving HREC(s), and the TGA independently reserve the right to discontinue the 
trial at any time for safety or other reasons. This will be done in consultation with the Sponsor where 
practical. In the event of premature trial termination or suspension, the above-mentioned parties will be 
notified in writing by the terminator/suspender stating the reasons for early termination or suspension 
(with the exception of the TGA as it is the Sponsor’s responsibility to notify regulatory authorities). After 
a decision to prematurely terminate the trial or to suspend the trial, the Sponsor and the PI will ensure 
that adequate consideration is given to protecting the participants’ interest and safety. The PI must review 
all participants as soon as practicable and complete all required records.  
7.1.1 CRITERIA FOR ADMINISTRATION OF RESCUE MEDICATION INSTEAD OF IMP DOSING 
ON DAY 8 
 
Once inoculated, it is expected that some study participants will experience malaria symptoms and 
biological changes (i.e. mild ALT/AST elevation, mild decrease in neutrophil count). Therefore, the 
following criteria should be considered as guidance for the decision not to proceed with pyronaridine 
dosing on Day 8 and preference for rescue medication instead:  
 
1. A participant experiences an SAE or ≥ CTCAE Grade 3 event (e.g. inoculum-related events such as 
lymphopenia or neutropenia). 
2. A participant has an ALT or AST > 2 x ULN or total bilirubin > 1 x ULN. 
3. A participant with positive alcohol or urine drug screen. 
4. A participant who tests positive for SARS-CoV-2 (section 8.3.7).  
5. A participant with one of the following ECG abnormalities: 
 
QTcF at any time >480 msec, 
 
Bundle branch block (except right bundle branch block that was present prior to pyronaridine 
administration), 
 
Any arrhythmia, except: 
a. Sinus bradycardia that is clinically asymptomatic, and not associated with any other 
relevant ECG abnormalities, 
b. sinus tachycardia that is clinically asymptomatic, and not associated with any other 
relevant ECG abnormalities, 
c. Respiratory sinus arrhythmia, 
d. Wandering atrial pacemaker, 


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37 
e. 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. 
6. The Investigator and delegates may also decide not to proceed with pyronaridine dosing based 
on other safety signals (clinical, ECG, vitals and laboratory tests) not included in the above criteria 
if there is a risk perceived for the study participants.   
In the event that participants are dosed with antimalarial rescue treatment instead of pyronaridine, 
participants will be administered their first dose of antimalarial rescue treatment at the clinic, but will 
then be able to return home. The participant will be contacted daily via phone to check that each dose of 
antimalarial treatment is being taken, and to ascertain any adverse events. Following antimalarial rescue 
treatment, blood will be taken for 18S qPCR until at least one negative result is obtained. No further bloods 
will be collected for drug concentration. Blood sampling for haematology and biochemistry and for future 
malaria research (if the participant has consented) will occur as scheduled until day 29±1. The EOS visit 
may be brought forward to day 29±1, at the discretion of the PI.   
If more than two discontinuations pre-IMP administration occur, additional participants may be recruited 
to subsequent cohorts to replace the discontinued participants on agreement with the study Sponsor. 
7.2 
VOLUNTEER DISCONTINUATION/WITHDRAWAL FROM THE STUDY 
Participants are free to withdraw from participation in the trial at any time upon request.  
An Investigator may discontinue or withdraw a participant from the trial for the following reasons: 
 
Pregnancy 
 
Significant trial intervention non-compliance  
 
If any clinical AE, laboratory abnormality, or other medical condition or situation occurs (or an 
exclusion criteria is revealed that was not previously recognised) such that continued participation 
in the trial would not be in the best interest of the participant 
The reason for participant discontinuation or withdrawal from the trial will be recorded on the eCRF. 
Participants who sign the informed consent form and are randomised but do not receive malaria 
inoculation may be replaced. Participants who sign the informed consent form, and are randomised and 
receive malaria inoculation, and subsequently withdraw, or are withdrawn or discontinued from the trial, 
may be replaced after mutual agreement between the Sponsor and the PI. The decision regarding the 
replacement of participants will be documented. 
Participants who have been inoculated and indicate they wish to withdraw from the trial must complete 
the full course of definitive antimalarial treatment. 
Participants will be reminded of the importance of completing their antimalarial treatment. Additionally, 
the PI will demonstrate due diligence in following up with the participant to ensure antimalarial therapy 
has been completed successfully. Participants may also be treated with Primacin™ if gametocytaemia is 
suspected from parasite lifecycle qRT-PCR or by the presence of stable low-level parasitaemia to ensure 
complete clearance of gametocytes. The decision to administer Primacin™ will be made by the PI in 
consultation with the medical monitor.  


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38 
If a participant is withdrawn from the trial, the Sponsor will be informed immediately. If there is a medical 
reason for withdrawal, the participant will remain under the supervision of the PI until satisfactory health 
has returned, or medical/clinical care has been transferred to the participant’s general practitioner or to 
a hospital consultant. 
The PI will make every effort to determine the primary reason for a participant’s withdrawal from the trial 
and record this information in the eCRF. If the participant is withdrawn from the trial procedures or follow-
up for any reason, with the participant’s permission, medical care will be provided for any SAEs that 
occurred during participation in the trial until the symptoms of any SAEs are resolved and the participant’s 
condition becomes stable. 
7.3 
LOST TO FOLLOW-UP 
If a participant does not attend a scheduled visit, the Investigator will apply due diligence by documenting 
all steps taken to contact the participant (e.g., dates of phone calls, registered letter, home visit, etc.) in 
the source documents. 
It will be explained to the participants during the consenting process and throughout the trial that they 
must be readily contactable. The period from inoculation to definitive antimalarial treatment is the critical 
period for participant safety because they may have blood stage parasites. A participant will be deemed 
“missing” if they fail to reply to communication to their personal mobile phone and nominated contact’s 
number after 24 h. If after 36 h the participant fails to respond then an Investigator will organise a home 
visit. Subsequently, if the participant is still absent, an Investigator will request assistance from the local 
police to locate the missing participant. Once the participant is found, the participant will be administered 
Riamet if this has not already been administered®, and Primacin® if required. 
A participant will be deemed lost to follow-up in the unlikely event that the participant is unable to be 
contacted or located despite all of the above measures. This is not anticipated to occur prior to 
administration of antimalarial rescue treatment, as in this situation, all efforts to locate the participant, 
including with assistance from local police, would continue until the participant is located and treated. 
8 
STUDY ASSESSMENTS AND PROCEDURES 
8.1 
STUDY CONDUCT SCHEDULE 
8.1.1 SCREENING VISIT (DAY -28 TO DAY -1)  
A screening visit will be scheduled after an initial phone interview conducted by clinical trial unit staff has 
occurred to review background information. For the screening visit, potential participants will be asked to 
come to the clinical trial unit after an overnight fast of ≥8 hours. During this initial screening visit, an 
Investigator will discuss the details of the trial with the potential participant, and the participant will read 
the Participant Information Sheet and be encouraged to ask questions. The potential participant will be 
fully informed of the nature of the trial at this time. Individuals willing to be considered for inclusion may 


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39 
sign the Informed Consent Form during the screening visit or may return to the clinical trial unit after 
further consideration of the trial and the Informed Consent Form. The participant will be given a copy of 
the Participant Information Sheet and signed Informed Consent Form for their records. The consenting 
process will be documented in the source notes. 
The signed and dated originals will be held on file by the clinical trial unit. Participation consent must be 
obtained from all individuals prior to screening tests. Once the potential participant has provided written 
consent to participate, the pre-trial screening may be initiated. 
Participants who complete all screening procedures and satisfy all entry criteria will be considered eligible 
to participate in this trial. For eligibility parameters, 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. In case of doubt, the PI is to confer with the Medical Monitor for agreement.   
If screening laboratory results are abnormal (e.g. HIV testing), the participant will be referred for 
appropriate counselling. If any clinically significant abnormalities are detected during screening, the 
participant will be referred to a general practitioner or medical specialist for follow-up tests as 
appropriate. 
Participants will be advised of the requirement to repeat some screening tests during the Day -3 to Day -
1 eligibility confirmation visit (unless screening occurs within eligibility Day -3 to -1 window) and/or on 
inoculation day to determine their continuing eligibility.  
The screening will be conducted within 28 days prior to the inoculation day and will include all activities 
listed below: 
 
A screening number will be assigned to each participant (as per clinical trial unit standard format). 
 
Elicit a complete medical history and use of medications.  
 
Elicit a social history including recreational drug, alcohol and tobacco use. 
 
Elicit demographic data.  
 
Record body height and weight. 
 
Perform alcohol breath test. 
 
Perform urine drug screen.  
 
Perform complete physical examination. 
 
Ask participant to complete the Beck Depression Inventory. 
 
Assess the cardiovascular disease risk (as per the Australian Absolute Cardiovascular Disease Risk 
Calculator). 
 
Record vital signs (supine and standing blood pressure and heart rate). 
 
Obtain a 12-lead ECG.  
 
Collect urine for urinalysis. 
 
Collect NPA for SARS-CoV-2 testing (if screening visit is conducted two days prior to inoculation). 
 
Collect blood samples for haematology and biochemistry (including lipids for cardiovascular risk 
factor), red cell antibodies, quantitative G6PD, serology and coagulation profile. 
 
Perform serum β-hCG pregnancy test for all females, and FSH test for post-menopausal females. 


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40 
 
Verify participant meets inclusion/exclusion criteria. 
8.1.2 ELIGIBILITY CONFIRMATION VISIT (DAY -3 TO DAY -1)  
The participants (including reserve participants) will report to the clinical trial unit between Day -3 to Day 
-1 for the following baseline assessments, unless screening laboratory assessments were conducted 
within this period, in which case repeat sampling will not be required.  
This visit will include: 
 
Collect blood samples for haematology and biochemistry. 
 
Collect urine for urinalysis. 
 
Collect NPA for SARS-CoV-2 testing. 
The timing of these assessments is to ensure that results are available for review by the Investigator prior 
to inoculation on Day 0.  
8.1.3 MALARIA CHALLENGE AGENT ADMINISTRATION (DAY 0)  
Participants, including up to three reserve participants per inoculation day, will report to the clinical trial 
unit on the morning of the day of inoculation. The Investigator will review the participants’ screening and 
eligibility confirmation visit results prior to their inoculation.  
On admission to the clinical trial unit, participants will be required to undertake further procedures to 
determine whether they remain eligible to be enrolled. A reserve participant may be asked to replace a 
participant who does not continue to meet eligibility. The reserve participants will be compensated for 
the study visit even if not inoculated, as described in the Participant Information Sheet and Consent Form.  
The procedures that will be undertaken prior to inoculation include: 
 
Verify that all applicable eligibility criteria have been met.  
 
Elicit information regarding any new medical conditions, illnesses and medication use since screening 
visit. 
 
Perform RAT for SARS-CoV-2 testing  
 
Perform alcohol breath test. 
 
Perform urine drug screen.  
 
Perform urine β-hCG pregnancy test for WOCBP. 
 
Perform abbreviated physical examination.  
 
Record vital signs  
 
Obtain a 12-lead ECG.  
 
Cannulate participants with an indwelling intravenous cannula (20 or 22 gauge) for the malaria 
parasite challenge agent, and record which arm is utilised.  
 
Collect blood samples for;  
o malaria 18S qPCR (baseline sample),  
o safety serum retention sample, 


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41 
o malaria future research sample (if participant has consented) 
Administration of the malaria challenge agent: 
 
Administer the malaria challenge agent of ~2,800 viable P. falciparum 3D7 infected human 
erythrocytes intravenously.   
 
Observe for a minimum of 60 minutes after inoculation to evaluate for immediate adverse reactions.  
 
Educate participants on signs and symptoms of malaria.  
a. Signs of malaria include fever (oral temperature above 38°C), chills/shivering/rigors, 
tachycardia, hypotension 
b. Symptoms of malaria include headache, myalgia, arthralgia, fatigue/lethargy, malaise, 
sweating/hot spells, loss of appetite, nausea, vomiting and abdominal discomfort. 
 
Provide participants with diary cards and thermometers to record any temperature readings during 
the study in the event of symptoms of fever. Participants will also record symptoms, details of self-
administered take home antimalarial rescue medication and concomitant medications on the diary 
cards during the study.  
 
Record AEs and concomitant medications.  
 
Record vital signs prior to leaving the clinical trial unit (approximately 60 minutes after inoculation). 
8.1.4 OUTPATIENT MONITORING PRE-IMP ADMINISTRATION (DAY 1 TO DAY 7)  
Monitoring via phone (Days 1 to 3): 
During this period, the participants are expected to be asymptomatic. A daily phone call or text message 
will be made to the participant by clinical trial unit staff to monitor participant well-being and to solicit 
any AEs and concomitant medications. 
Daily visits to the clinical trial unit for monitoring (Days 4 to 7):  
Follow-up from Day 4 to Day 7 will be undertaken through daily morning visits to the clinical trial unit. 
The following procedures will occur during these visits:  
 
Perform symptom-directed physical examination if clinically indicated.  
 
Record vital signs.  
 
Collect blood sample for malaria 18S qPCR. 
 
Collect NPA for SARS-CoV-2 testing (Day 6 am) 
 
Collect blood samples for haematology and biochemistry (Day 7 am)  
 
Record AEs and use of concomitant medications.  
 
Check participant diary cards. 
8.1.5 INPATIENT OBSERVATION AND IMP ADMINISTRATION (DAY 8 TO DAY 12)  
Participants will be admitted to the clinical trial unit for single-dose administration of pyronaridine on Day 
8 when parasitaemia for the majority of participants is expected to be above 5,000 parasites/mL.  


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42 
The following procedures will occur at admission to the clinical trial unit (before pyronaridine 
administration): 
 
Perform RAT for SARS-CoV-2 testing  
 
Perform abbreviated physical examination.  
 
Obtain and review a 12-lead ECG. 
 
Record vital signs. 
 
Perform and review urine β-hCG pregnancy test for WOCBP. 
 
Record AEs and use of concomitant medications. 
 
Perform and review alcohol breath test and urine drug screen. 
 
Review haematology and biochemistry results from Day 7 am collection.  
 
Cannulate participants with an indwelling intravenous (18 or 20 gauge) cannula.  
 
Collect blood samples for  
o haematology and biochemistry,  
o malaria 18S qPCR,  
o parasite viability ex vivo growth, 
o drug concentration (baseline sample), 
o malaria future research sample (if participant consented)  
The following procedures will occur after pyronaridine administration: 
 
Administer the relevant pyronaridine dose under direct observation. 
 
Follow up participants as in-patients for up to 96 hours to ensure tolerance of pyronaridine and 
adequate clinical response. 
 
Perform symptom-directed physical examination if clinically indicated. 
 
Record vital signs three times a day (approximately 6 – 8 hours apart) whilst confined.  
 
Obtain 12-lead ECG once daily while confined. 
 
Collect blood for haematology and biochemistry at 72 hours post-pyronaridine administration. 
 
Collect blood for malaria 18S qPCR following pyronaridine administration at 4, 8, 12, 16, 20, 24, 30, 
36, 42, 48, 60, 72, 84 and 96 hours (see section 8.4 for time allowed time windows). 
 
Collect blood samples for drug concentration at 1, 2, 3, 4, 6, 8, 12, 16, 20, 24, 30, 36, 42, 48, 60, 72, 
84 and 96 hours (see section 8.4 for time allowed time windows).  
 
Collect blood for parasite viability ex vivo growth at 4, 8, 12, 16, 20, 24, 48, 72 and 96 hours (see 
section 8.4 for time allowed time windows). 
 
Record AEs and use of concomitant medications. 
Participants will be confined in the clinical unit for up to 96 hours to ensure tolerance of pyronaridine and 
adequate clinical response. In the event that parasitaemia is not reduced from peak parasitaemia value 
by approximately 10-fold during the confinement period, and/or parasite regrowth occurs (≥5000 
parasites/mL and a 2-fold parasitaemia increase within 48 hours), rescue treatment with Riamet® will be 
initiated.  
If participants are clinically well at the end of the 96 hour confinement, they will be discharged and 
monitored on an outpatient basis up to the EOS visit. Before exit from confinement, clinical trial unit 


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nursing staff must confirm with the Investigator if any participant requires symptom-directed physical 
examination prior to discharge. 
The following procedures will occur prior to discharge from the clinical trial unit (96 hours): 
 
Perform symptom-directed physical examination if clinically indicated. 
 
Obtain a 12-lead ECG. 
 
Record vital signs.  
 
Collect blood samples for  
o malaria 18S qPCR, 
o drug concentration, 
o parasite viability ex vivo growth 
 
Record AEs and use of concomitant medications. 
8.1.6 OUTPATIENT MONITORING POST-IMP ADMINISTRATION (DAY 13 TO DAY 50±2)  
Follow up visits will be undertaken on an out-patient basis for clinical evaluation and blood sampling. 
Follow up visits will occur at least three times per week (may occur more frequently if clinically indicated) 
until Riamet® treatment. 
The following procedures will take place during these visits: 
 
Obtain 12-lead ECG.  
 
Record vital signs.  
 
Record AEs and use of concomitant medications. 
 
Check participant diary cards. 
 
Perform symptom-directed physical examination if clinically indicated. 
 
Collect blood samples for drug concentration at 120 (Day 13±1), 168 (Day 15±1), 216 (Day 17±1), 288 
(Day 20±1), 336 (Day 22±1), 384 (Day 24±1), 456 (Day 27±1), 504 (Day 29±1), 576 (Day 32±2), 744 (Day 
39±2) and 864 (Day 44±2) hours (Days) post-dose 
 
Collect blood samples for malaria 18S qPCR at least 3 times per week until Riamet® treatment, timing 
will be at PI discretion based on previous 18S qPCR results and clinical symptoms. Where possible 18S 
qPCR blood sampling should coincide with drug concentration sampling time-points to avoid bringing 
participants back for outpatient visits unnecessarily.  
 
Collect blood samples for haematology and biochemistry on Days 15±1, 22±1, 29±1, 37±2 and 44±2.  
 
Collect blood samples for parasite lifecycle stage qRT-PCR at Investigator’s discretion (if 
gametocytaemia is suspected based on low and stable parasitaemia). These samples should be aligned 
with the 18S qPCR to avoid bringing participants back for outpatient visits unnecessarily.    
 
Collect blood samples for malaria future research (if participant has consented) on Days 15±1 and 
29±1.  
 
 


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8.1.7 ANTIMALARIAL RESCUE TREATMENTS 
All participants will receive compulsory antimalarial rescue treatment with Riamet® on Day 47±2, or earlier 
in the following cases: 
 
Parasitaemia is not reduced by approximately 10-fold at the end of the 96 hour confinement 
period (Day 12) when compared with peak parasitaemia. 
 
Parasite regrowth occurs after an initial reduction in parasitaemia, with regrowth defined as 
≥5000 parasites/mL and a 2-fold parasitaemia increase within 48 hours.  
 
Volunteer discontinuation/withdrawal from the study (see Section 7.2). 
 
Investigator’s discretion in the interest of participant safety. 
As qPCR results are not received in real-time, rescue may occur when these parasitaemia thresholds are 
presumed to have occurred. 
If gametocytes are present as determined by qRT-PCR at any time point after the confinement period, a 
single dose of Primacin® will be administered at the Investigator’s discretion.  A participant’s G6PD status 
will determine the dose of primaquine administered. If an allergy or contraindication to Riamet® or 
Primacin® develops, Malarone® will be administered. In the rare event that a participant cannot tolerate 
oral drugs, participants will be admitted to hospital and I.V. artesunate will be administered.  
The following procedures will be performed prior to antimalarial rescue treatment: 
 
Perform symptom-directed physical examination if clinically indicated. 
 
Obtain a 12-lead ECG. 
 
Record vital signs. 
 
Record AEs and use of concomitant medications. 
 
Check participant diary cards. 
 
Collect blood samples for  
o haematology and biochemistry,  
o malaria 18S qPCR, 
o drug concentration 
 
Collect sample for parasite viability ex vivo growth. NOTE only if antimalarial rescue treatment is 
administered prior to Day 47±2.   
Please note if a participant receives antimalarial rescue treatment after administration of pyronaridine 
but prior to Day 47±2, drug concentration, haematology, biochemistry and future malaria research blood 
collection will still be required for the full duration of the trial. Following antimalarial rescue treatment 
blood will be taken for 18S qPCR at the completion of Riamet and until at least one negative result is 
obtained.  
 
 


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45 
8.1.8 END OF STUDY VISIT (DAY 50±2) 
The EOS visit must occur at a minimum 3-days after commencement of compulsory antimalarial 
treatment. 
The following procedures will be performed at the EOS visit: 
 
Perform a complete physical examination. 
 
Obtain a 12-lead ECG. 
 
Record vital signs. 
 
Record body weight. 
 
Record AEs and use of concomitant medications. 
 
Collect urine for urinalysis.  
 
Collect blood samples for 
o haematology and biochemistry,  
o serum β-hCG pregnancy test for WOCBP, 
o RBC alloantibodies,  
o serology, 
o malaria 18S qPCR, 
o drug concentration, 
o safety serum retention sample, 
o malaria future research sample (if participant consented) 
 
Collect diary card. 
If a participant is unwell with detectable parasitaemia or at the Investigator’s discretion, the participant 
may be asked to return to the clinical trial unit after the EOS visit for safety follow up. 
8.2 
PHARMACOKINETIC AND PHARMACODYNAMIC ASSESSMENTS  
8.2.1 PHARMACOKINETICS 
Blood will be collected to quantify whole blood pyronaridine concentrations for PK analysis. PK data will 
be used to calculate the PK/PD parameters of pyronaridine using a population PK model (the primary 
endpoint). PK parameters of pyronaridine will also be calculated using non-compartmental methods 
(secondary endpoint). 
Samples will be analysed for pyronaridine blood concentrations using a validated analytical method in 
compliance with the SOPs of the laboratory performing the analyses. Assays will be performed in a 
Sponsor approved laboratory. 
Blood for pyronaridine concentrations will be collected pre-pyronaridine administration on Day 8, then at 
the following hours post-pyronaridine administration while confined in the clinical unit: 1, 2, 3, 4, 6, 8, 12, 
16, 20, 24, 30, 36, 42, 48, 60, 72, 84 and 96 hours. As outpatients, participants will have blood collected 


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46 
on the following days: 13, 15, 17, 20, 22, 24, 27, 29, 32, 39, 44, 47 and 50. Allowed time windows for 
sample collection are specified in Section 8.4.  
8.2.2 PHARMACODYNAMICS 
Malaria 18S qPCR  
Blood will be collected to quantify total malaria parasitaemia by quantitative polymerase chain reaction 
targeting the gene encoding P. falciparum 18S rRNA (malaria 18S qPCR; reported as parasites/500uL 
packed RBCs). Malaria 18S qPCR data will be used to calculate the PK/PD parameters of pyronaridine (the 
primary endpoint) and to determine the kinetics of parasite clearance following pyronaridine dosing 
(secondary endpoint). Malaria 18S qPCR will also be used to monitor the parasitaemia of participants in 
real time in order to guide the timing of antimalarial rescue treatment initiation.  
Samples for malaria 18S qPCR will be analysed using a validated analytical method in compliance with the 
SOPs of the laboratory performing the analyses. Assays will be performed in a Sponsor approved 
laboratory. 
Blood for malaria 18S qPCR will be collected on Day 0 prior to inoculation with the malaria challenge agent, 
then once daily on Days 4, 5, 6 and 7. On Day 8, blood will be collected pre-pyronaridine administration, 
then at the following hours post-pyronaridine administration while confined in the clinical unit: 4, 8, 12, 
16, 20, 24, 30, 36, 42, 48, 60, 72, 84 and 96 hours.  
As outpatients, participants will have blood collected at least three times per week until antimalarial 
rescue treatment. Timing will be at PI discretion based on previous malaria 18S qPCR results and clinical 
symptoms. Where possible, malaria 18S qPCR blood sampling should coincide with drug concentration 
sampling time-points to avoid unnecessary outpatient visits. Following antimalarial rescue treatment, no 
further blood collection for malaria 18S qPCR is required for a participant once a minimum of two negative 
results are recorded.  
Allowed time windows for sample collection are specified in Section 8.4.  
Parasite lifecycle stage qRT-PCR 
Additional blood samples may be collected to evaluate the presence of sexual parasite stages 
(gametocytes) and other parasite lifecycle stages using quantitative reverse-transcriptase polymerase 
chain reaction (qRT-PCR). The qRT-PCR assays may target parasite transcripts including but not limited to: 
the female gametocyte-specific transcript pfs25, the male gametocyte-specific transcript PfMGET, and the 
ring-stage transcript pfSBP-1 as appropriate. The samples may be taken at time points indicated by malaria 
18S qPCR, or at the Investigator’s discretion. The parasite lifecycle stage qRT-PCR data may be used to 
facilitate the calculation of the PK/PD parameters of pyronaridine (the primary endpoint). Further, the 
results may guide the requirement for Primacin® administration to participants prior to the end of study.  
 


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47 
Parasite viability in ex vivo culture 
Ex vivo growth is an indirect measure of parasite viability. While 18S qPCR quantifies all parasites 
(including those non-viable or dead in circulation), only viable parasites are able to replicate in ex vivo 
culture. Modelling of parasite growth observed ex vivo enables the fraction of viable parasites present in 
blood samples to be determined. The results of parasite ex vivo growth analyses will be supportive to 
malaria 18S qPCR data in determining the kinetics of parasite clearance following pyronaridine dosing 
(secondary endpoint).  
Short term ex vivo cultures will be established from blood collected pre-pyronaridine administration (Day 
8), then at the following hours post-pyronaridine administration while confined in the clinical unit: 4, 8, 
12, 16, 20, 24, 48, 72 and 96. An additional blood sample will be collected prior to the initiation of Riamet® 
treatment in the event that treatment is initiated prior to Day 47±2.  
 
Microscopic examination for evidence of parasitaemia or gametocytaemia may also be conducted at the 
Investigator’s discretion. 
Refer to the Laboratory Manual for sample preparation procedures and sample volumes. 
8.3 
SAFETY AND OTHER ASSESSMENTS 
The schedule for all safety assessments is summarised in the SOA table (section 1.3) and in more detail in 
the study conduct schedule (Section 8.1). All safety assessments may be conducted at unscheduled visits 
or time points if required for the participant’s safety at the discretion of the Investigator. In the case of 
elevated AST, ALT or total bilirubin, safety assessments will occur twice a week until resolution.  
When several assessments are scheduled to take place at the same time point, the preferred order is: 
ECG, vital signs, blood sampling, urine sampling. When the malaria challenge agent, pyronaridine, and 
rescue medication are administered, procedures that should occur pre and post administration are 
detailed in Section 8.1.   
Exact timing of blood sampling to measure pyronaridine blood concentrations and to monitor malaria 
parasitaemia is to be maintained as closely as possible and therefore safety assessments scheduled for 
these time-points should be conducted ahead of the time-point. Please refer to the allowed time windows 
(Section 8.4).  
 
8.3.1 BECK DEPRESSION INVENTORY 
Originally described by Beck et al (1961), the Beck Depression Inventory (BDI) is a validated objective tool 
for the assessment of depression. Updated in 1996, the BDI-II is a 21-item, self-report rating inventory 
that measures characteristic attitudes and symptoms of depression, and participants will be required to 
complete the BDI-II at Screening for eligibility. The BDI-II takes approximately 10 minutes to complete, 
although participants require a fifth – sixth grade reading level to adequately understand the questions. 


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48 
A score of >20 at screening and/or a response of 1, 2 or 3 for item 9 indicating current suicidal ideation is 
exclusionary. A participant with a BDI-II score of 17 to 19 may be enrolled at the discretion an Investigator 
if they do not have a history of the psychiatric conditions mentioned in exclusion criterion 10-12 and their 
mental state is not considered to pose additional risk to the health of the participant or to the execution 
of the trial and interpretation of the data gathered. The results will be entered into the eCRF.  
8.3.2 PHYSICAL EXAMINATION  
A full physical examination includes the following: 
Weight  
Height (Screening only)  
Review of systems excluding genitourinary examination and including the following:  
Head, neck (including thyroid), ears, eyes, nose and throat  
Heart/circulation  
Chest  
Lungs  
Abdomen  
Skin  
Neurological exam  
A Full physical examination will be performed at Screening and at EOS.  
An abbreviated physical examination will include heart/circulation, chest, lungs, skin, and abdomen. An 
abbreviated physical examination will performed prior to inoculation with the malaria challenge agent 
and upon admission to the clinical trial unit on Day 8.  
When the symptom-directed physical examination is performed, body systems will be reviewed only if 
clinically indicated. 
All physical examinations after inoculation with the malaria challenge agent on Day 0, the following signs 
and symptoms of malaria infection should be considered: 
 
Signs of malaria include fever (oral temperature above 38°C), chills/shivering/rigors, tachycardia, 
hypotension 
 
Symptoms of malaria include headache, myalgia, arthralgia, fatigue/lethargy, malaise, sweating/hot 
spells, loss of appetite, nausea, vomiting and abdominal discomfort. 
8.3.3 BODY MEASUREMENTS 
Body weight (kg) and height (cm) will be measured at screening. Body weight (kg) will also be measured 
at the EOS visit.  
8.3.4 VITAL SIGNS 


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49 
Vital signs (temperature, heart rate, respiratory rate, and blood pressure) will be measured after the 
participant has rested in the supine position for at least 5 min and in the standing position within 3 min 
when changing from the supine to standing position (blood pressure and heart rate only) at Screening.  
At all other time points, vital signs will be measured after the participant has rested in the seated position 
for at least 5 min.  
The normal ranges for vital signs once on trial are: 
Parameter 
Range 
Systolic blood pressure             
90-140 mmHg 
Diastolic blood pressure           
50-90 mmHg 
Heart rate                                 
50-100 bpm 
Temperature                             
35.0-37.5°C 
Respiratory rate                        
10-25 breaths/min 
8.3.5 12-LEAD ELECTROCARDIOGRAM (ECG) 
A 12-lead ECG will be recorded after the participant has rested supine for at least 5 min. ECG tracings will 
be retained and labelled as per standard procedures at the clinical trial unit and will be recorded in the 
eCRF. Any clinically significant findings will be discussed with the Medical Monitor and Sponsor and 
documented as AEs. An Investigator will sign and date each ECG as evidence of their review. 
The normal ECG ranges once on trial are: 
Parameter 
Range 
PR interval             
≤210 msec 
QRS           
50–120 msec 
QTcF                            
Males: ≤450 msec 
Females: ≤470 msec 
8.3.6 LABORATORY TESTS 
The required pathology tube types and blood and urine volumes required for each procedure and 
laboratory processing procedures will be described in the Laboratory Manual.  
Any significant deviations from results obtained during screening will be followed until resolution or 
investigated fully, or until the participant is referred to a general practitioner or medical specialist as 
appropriate. An Investigator will document the clinical significance of all results falling outside of the 
normal reference ranges.  
Blood and urine samples for clinical laboratory assessments will be collected according to clinical trial unit 
SOPs. The parameters that will be measured are listed below. Additional reflex testing may be conducted 
by the local laboratory (as per their SOPs) if safety laboratory values for a participant fall outside of the 
normal range/parameters. Unscheduled testing may be performed at Investigator’s discretion. 


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Blood will be collected for clinical laboratory evaluations including haematology, biochemistry, 
coagulation profile, serology, G6PD, pregnancy testing or FSH testing, and for red cell antibodies. All safety 
samples will be sent to national laboratories as named in this protocol (section 10.1.5). 
8.3.6.1 HAEMATOLOGY 
 
Full blood count (FBC) with differential  
White blood cell count (WBC)  
WBC differential (diff)  
A manual blood smear should be reviewed if there are immature/abnormal cells detected on the 
automated differential or if an automated differential was not able to be performed.  
Neutrophils (NEUT) 
Lymphocytes (LYM) 
Monocytes (MON) 
Eosinophils (EOS) 
Basophils (BAS) 
Mean corpuscular volume (MCV) 
Red blood cell count (RBC) 
Haemoglobin (HGB)  
Haematocrit (HCT)  
Platelet count (PLAT)  
Red blood cell distribution width (RDW) 
Reticulocyte count (RETI)  
Blood Group and Rh(D) tests (Screening only) 
8.3.6.2 BIOCHEMISTRY 
Sodium (NA) 
Potassium (K) 
Chloride (CL) 
Bicarbonate (BICARB) 
Calcium (CA) 
Corrected calcium (CCA) 
Magnesium (MAG) 
Glucose (GLUC) 
Urea 
Creatinine (CREAT) 
Estimated glomerular filtration rate (eGFR) – CKD Epi calculation 
Albumin (ALB) 
Globulin 


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Total protein 
Total bilirubin (BILI) 
Direct (conjugated) bilirubin (BILDIR) 
Alkaline phosphatase (ALP) 
Alanine aminotransferase (ALT) 
Aspartate aminotransferase (AST) 
Gamma-glutamyl transferase (GGT) 
Lactate dehydrogenase (LDH) 
Phosphate (PHOS) 
C-Reactive Protein (CRP)  
Cholesterol (Chol) 
Triglycerides (Trig) 
8.3.6.3 LIPIDS  
Total cholesterol 
High density lipoprotein (HDL) 
Low density lipoprotein (LDL) 
Triglycerides 
8.3.6.4 SEROLOGY 
HIV 1/2 (anti-HIV1 and anti-HIV2 Ab) 
Hepatitis B (HBsAg, anti-HBc [IgG + IgM if IgG is positive]) 
Hepatitis C (anti-HCV) 
8.3.6.5 COAGULATION 
Blood for Prothrombin Time (PT), Activated Partial Thromboplastin Time (APTT) and International 
Normalised Ratio (INR) testing will be collected at screening. 
8.3.6.6 RBC ALLOANTIBODIES AND G6PD TESTING 
Blood for red cell antibody testing will be collected for participants at screening and at the EOS visit.  
Blood for G6PD testing will be collected at screening only.  
8.3.6.7 PREGNANCY TESTING 
At screening and at the EOS visit a serum β-hCG pregnancy test will be conducted for all female 
participants, and FSH levels will be tested in post-menopausal females (at least one year post-
menopausal). On Days 0 and 8, a urine β-hCG pregnancy tests will be conducted in WOCBP. If a participant 


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52 
is found to have a positive urine pregnancy test at any stage during the study, a confirmatory serum β-
hCG will be performed. If confirmed positive, the participant will be discontinued from the study and 
rescued with Riamet®. Please refer to Section 8.5.9 for pregnancy-specific reporting procedures and 
follow-up.  
8.3.6.8 URINALYSIS 
Urine will be tested by dipstick at the clinical trial unit. If there are any abnormalities considered clinically 
significant in blood, leukocytes, or protein, the urine will be sent for formal laboratory urinalysis per the 
clinical trial unit standard procedure for microscopy, culture and sensitivity. 
Glucose (GLUC) 
Bilirubin (BILI) 
Ketone (KETONES) 
Specific gravity (SPGRAV) 
Blood  
pH  
Protein (PROT) 
Urobilinogen (UROBIL) 
Nitrite  
Leukocytes (WBC)  
Microscopy, culture and sensitivity (if required)  
8.3.6.9 URINE DRUG SCREEN AND ALCOHOL BREATH TESTS 
All participants will be questioned about concomitant medications and use of recreational drugs. The 
urine drug screen may be repeated if the potential participant denies usage of any of these agents and 
the test result is believed to be a false positive. 
Urine drug screen: 
Amphetamines 
Opiates 
Methamphetamines 
Phencyclidine 
Barbiturates 
Tetrahydrocannabinol (cannabis) 
Benzodiazepines 
Tricyclic antidepressants 
Cocaine  
 
Methadone 
 
Alcohol breath test 
If the results of the urine drug screens or alcohol breath tests are positive, participants will not be enrolled 
or administered pyronaridine on Day 8 unless there is an explanation acceptable to an Investigator (e.g., 
the participant has stated in advance that they consumed a prescription or over-the-counter product that 
contained the detected drug) and the participant has a negative urine drug screen on retest by the 
pathology laboratory. 


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8.3.7 SARS-COV-2 RISK MANAGEMENT: 
 
A NPA will be conducted at the eligibility visit or screening visit if it falls within the eligibility window (SoA). 
A RAT may be conducted in place of PCR, if PCR is not available or if results are not expected back within 
the required timeframe. Participants who test positive for SARS-CoV-2 at this time-point will be excluded 
from the study, and will be managed as per Queensland Health guidelines for management of COVID-19 
positive individuals. 
All eligible participants will be required to have had a negative Rapid Antigen Test (RAT) immediately prior 
to (i.e., that morning) being admitted to the clinical trial site for inoculation with the malaria challenge 
agent (Day 0). If any participant tests positive for SARS-CoV-2 at this time-point they will be excluded from 
the study, and will be managed as per Queensland Health guidelines for management of COVID-19 
positive individuals.  
For participants who are inoculated with the malaria challenge agent, a repeat NPA for SARS-CoV-2 by 
PCR will be conducted on Day 6 (a RAT may be conducted in place of PCR, if PCR is not available or if results 
not expected back within the required timeframe).  In addition, a RAT will be conducted prior to 
confinement on the morning of Day 8.  If any participant tests positive at any time point before or after 
confinement they will be managed as per Queensland Health guidelines for management of COVID-19 
positive individuals. Definitive antimalarial treatment will be delivered to the participants’ home address, 
and a phone call will be made to ensure that the first dose is taken; additional daily phone contact will be 
made to ascertain adverse events and to ensure that all subsequent doses are taken. No further clinic 
visits will be conducted during the time that the participant is required to isolate; however, where 
possible an EOS visit will be conducted following release from isolation.  
In the event that a participant tests negative by RAT on the morning of confinement (Day 8), but then 
tests positive by RAT during confinement, the participant will be released from the clinical trial site to 
isolate at home. Participants will leave the clinical site with the full course of definitive antimalarial 
medication. Daily phone contact will be made to ascertain adverse events and to ensure that all doses of 
antimalarial medication are taken. If a participant’s condition was to deteriorate while isolating at home, 
the participant will be assessed over the phone by an Investigator and may be advised to present to a 
hospital Emergency Department if required, or clinical site staff may call 000 if appropriate. In the event 
that a participant is admitted to hospital, the PI will continue to liaise with treating clinicians. Study visits 
will resume where possible following the participant's release from isolation. All other participants still 
confined at the clinical trial site will undergo daily RAT testing for the duration of their confinement. 
 
8.3.8 SAFETY SERUM RETENTION SAMPLE 
A 5 mL blood sample will be collected on Day 0 prior to inoculation with the malaria challenge agent and 
at EOS.  
Participants consent to this mandatory collection and storage and the use of the sample for safety 
assessments when they sign the Informed Consent Form for the study. Safety serum samples will be 
retained for at least 15 years from the completion of the study at the QIMR Berghofer Institute. 


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8.3.9 MALARIA FUTURE RESEARCH 
Blood samples may be collected for storage and use in future malaria research under a separate protocol 
and consenting process. The future research will be under the local sponsorship of QIMR Berghofer 
Medical Research Institute and will involve a protocol and consent document approved by the QIMR 
Berghofer HREC.  
If a participant consents to the collection of blood samples for future malaria research, up to six blood 
samples will be collected at time points that coincide with existing visits, including time points prior to 
and after administration of the malaria challenge agent. 
Blood volume of each sample may be up to and not exceeding 30 mL, and will be appropriate for the 
day/time point. The total volume of blood taken including the main study samples will not exceed 470 mL 
per month. 
A participant may change their decision to provide samples for future malaria research at any time prior 
to the EOS by notifying QIMR Berghofer, as local Sponsor, in writing. However, if a participant initially 
consents to future malaria research some of their blood has already been used for research purposes, the 
information from that research may still be used. 
8.4 
ALLOWED TIME WINDOWS 
The following time windows will be allowed for vital signs recording, ECGs and blood sampling for clinical 
laboratory evaluations: 
Time point 
Allowed time window 
Malaria inoculation (Day 0) 
± 4 hours 
Outpatient monitoring pre-IMP (Days 4-7) 
± 12 hours 
Inpatient observation pre-IMP (Day 8) 
± 2 hours 
Inpatient observation ≤24 hours post-IMP (Day 8-9) 
± 30 minutes 
Inpatient observation >24 hours post-IMP until discharge (Day 9-12) 
± 60 minutes 
Outpatient monitoring from discharge up to and including Day 30 
± 24 hours 
Outpatient monitoring from Day 31 until EOS visit 
± 2 days 
The following time windows will be allowed for blood sampling to quantify pyronaridine blood 
concentrations and to monitor malaria parasitaemia: 
Time point 
Allowed time window 
Malaria inoculation (Day 0) 
± 4 hours 
Outpatient monitoring pre-IMP (Days 4-7) 
± 12 hours 
Inpatient observation pre-IMP (Day 8) 
± 60 minutes 
Inpatient observation ≤24 hours post-IMP (Day 8-9) 
± 30 minutes 
Inpatient observation >24 hours post-IMP until discharge (Day 9-12) 
± 60 minutes 
Outpatient monitoring from discharge up to and including Day 30 
± 24 hours 
Outpatient monitoring from Day 31 until EOS visit 
± 2 days 


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8.5 
ADVERSE EVENTS AND SERIOUS ADVERSE EVENTS 
8.5.1 DEFINITION OF ADVERSE EVENTS (AE) 
An AE is any untoward medical occurrence, i.e., unfavourable and unintended sign (including an abnormal 
laboratory finding), symptom or disease that occurs in a participant during the course of the trial. An AE 
does not necessarily have a causal relationship with trial treatments or procedures.  
AEs include, but are not limited to: 
 
A new symptom, sign or medical condition. 
 
A disease or medical condition detected or diagnosed during the course of the trial even though it 
may have been present prior to the start of the trial. 
 
An exacerbation of a pre-existing medical condition/disease. 
 
An increase in frequency or intensity of a pre-existing episodic disease or medical condition. 
 
Continuous persistent disease or symptoms present at trial start that worsen following the start of 
the trial. 
 
An abnormal assessment (e.g., change on physical examination, ECG finding) if it represents a clinically 
significant finding that was not present at trial start or worsened during the course of the trial. 
 
An abnormal laboratory test result if it represents a clinically significant finding (e.g., CTCAE grade 2 
or above), symptomatic or not, which was not present at trial start or worsened during the course of 
the trial or led to dose reduction, interruption or permanent discontinuation of trial treatment. 
Borderline abnormal laboratory findings and other objective assessments should NOT be routinely 
captured and reported as AEs. However, abnormal laboratory findings or other objective measurements 
that meet the following criteria should be captured and reported in the AE section of the eCRF: 
 
The result meets the criteria for reporting as an SAE. 
 
The test result is associated with accompanying symptoms, and/or 
 
It requires additional diagnostic testing or medical/surgical intervention, and/or 
 
It leads to a change in trial dosing, or discontinuation from the trial, significant additional concomitant 
drug treatment, or other therapy, and/or 
 
It is considered by an Investigator to be clinically significant or represent a clinically significant change 
from baseline. 
Merely repeating an abnormal test, in the absence of any of the above conditions, does not constitute an 
AE. Any abnormal test result that is determined to be an error does not require reporting as an AE. Surgical 
procedures themselves are not AEs; they are therapeutic measures for conditions that may, or may not, 
be AEs.  Malaria in itself is not considered an AE. 
8.5.2 DEFINITION OF SERIOUS ADVERSE EVENTS (SAE)  
An SAE is defined as an event that fulfils at least one of the following criteria: 
 
Results in death. 
 
Is life-threatening  


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o The term "life-threatening" in the definition of "serious" refers to an event in which the participant 
was at immediate risk of death at the time of the event; it does not refer to an event which 
hypothetically might have caused death if it was more severe. 
 
Requires inpatient hospitalisation or prolongs existing hospitalisation, unless this is for: 
o 
Elective or pre-planned treatment for a pre-existing condition that is unrelated to the trial and 
has not worsened since the start of the trial. 
o 
Cosmetic surgery, or for social reasons, or respite care in the absence of any deterioration in the 
participant’s general condition. 
 
Results in persistent or significant disability/incapacity. 
 
Is a congenital abnormality/birth defect. 
 
Is considered medically important  
o 
Medical and scientific judgement should be exercised in deciding whether other AEs are to be 
considered serious, such as important medical events that may not be immediately life-
threatening but may jeopardise the participant or may require intervention to prevent one of the 
other outcomes listed in the definition above. Examples of such events are: intensive treatment 
in an emergency room or at home for allergic bronchospasm; blood dyscrasias; convulsions that 
do not result in hospitalisation; development of drug dependency or drug abuse. 
 
A Suspected Unexpected Serious Adverse Reaction (SUSAR) is any SAE where a causal relationship with 
a trial intervention (malaria challenge agent, pyronaridine, antimalarial rescue medication) is at least a 
reasonable possibility, and the event is not listed in the IB. 
8.5.3 DEFINITION OF EVENTS OF SPECIAL INTEREST  
An AE of special interest (AESI), whether serious or non-serious, is one of scientific and medical concern 
specific to the Sponsor’s product or program, for which ongoing monitoring and rapid communication by 
an Investigator to the Sponsor and medical monitor 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 trial Sponsor to other parties (e.g., regulators) might also be warranted 
(CIOMS VI, ICH E2F, 2010). Any abnormalities listed below should be reported as an AESI. 
Hepatic: 
 
Any ALT or AST value above 5×ULN. 
 
An elevation in bilirubin 2×ULN. 
 
Any AST or ALT value above 2×ULN and (total bilirubin >1.5×ULN). 
 
Any AST or ALT value above 2×ULN with the appearance of fatigue, nausea, vomiting, right upper 
quadrant pain or tenderness, fever, rash and/or eosinophilia (eosinophil percent or count above the 
ULN). 
 
Constitutes a possible Hy’s Law case 
o 
Hy’s Law case is defined as a participant with any value of alanine or aspartate aminotransferase 
greater than 3×ULN together with an increase in total bilirubin to a value greater than 2×ULN and 


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not associated to an alkaline phosphatase value greater than 2×ULN (FDA Guidance on Drug 
Induced Liver Injury: Premarketing Clinical Evaluation [2009]). 
 
Cardiac: 
 
QTcF at any time >480 msec. 
 
Bundle branch block (except right bundle branch block that was present prior to pyronaridine 
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, is 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, or 
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 
Haematological: 
 
Haemoglobin drop >20.0 g/L from baseline (at eligibility visit) 
 
Absolute neutrophil count (ANC) <0.5 × 109/L. 
 
Platelet count <75 × 109/L. 
Dermatological: *† 
Clinical signs of possible cutaneous adverse reactions (clearly related to malaria challenge agent 
inoculation and/or IMP administration; this does not include irritation from dressings and/or ECG dots) 
such as: 
 
Dermatitis 
 
Rash including 
o erythematous 
o macular 
o papular 
o maculopapular 
o pruritic 
o pustular 
o vesicular 
*If one of these cutaneous reactions is observed, pictures of the lesions should be obtained when feasible. 
†Dermatological AEs need not be reported as AESIs if clearly unrelated to the malaria challenge agent or 
IMPs (e.g. rash from cannula dressing or ECG dots). 
All AESIs, including those that do not meet the definition of an SAE, must be reported as per section 8.5.7 
 
 


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8.5.4 CLASSIFICATION OF AN ADVERSE EVENT 
8.5.4.1 SEVERITY OF EVENT 
In addition to determining whether an AE fulfils the criteria for an SAE or not, the severity of AEs 
experienced by participants will be recorded in accordance with the Common Terminology Criteria for 
Adverse Events (CTCAE) v5.0, published 27 November 2017. 
The severity of AEs will be graded as follows:  
• 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 living. 
• Grade 3: Severe or medically significant but not immediately life-threatening; hospitalisation or 
prolongation of hospitalisation indicated; disabling; limiting self-care activities of daily living. 
• Grade 4: Life-threatening consequences; urgent intervention indicated. 
• Grade 5: Death related to AE. 
A mild, moderate, or severe AE may or may not be serious. These terms are used to describe the intensity 
of a specific event. Medical judgement should be used on a case-by-case basis.  
Seriousness, rather than severity assessment, determines the regulatory reporting obligations. 
8.5.4.2 RELATIONSHIP TO STUDY INTERVENTION 
The PI or delegate must assess the relationship of each event to the malaria challenge agent, pyronaridine, 
and the antimalarial rescue medications (separately) and decide whether, in his or her medical judgement, 
there is a reasonable possibility that the event may have been caused by any of the trial interventions. 
Where possible, a distinction should be made between events considered related to the malaria challenge 
agent, pyronaridine, and the antimalarial rescue medications.  
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 cause-and-effect relationship between the 
malaria challenge agent, pyronaridine, or the antimalarial rescue medications and the occurrence of the 
AE (even if undetermined or untested), then the AE should be considered as “related” to whichever 
product is relevant. This should be documented in the participant’s clinical file (source data) and eCRF.  
The following may guide this assessment: 
Related/suspected:  
The temporal relationship between the event and the administration of the malaria challenge agent 
and/or pyronaridine and/or the antimalarial rescue medications 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.  


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Not related/not suspected:  
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 
challenge agent and/or pyronaridine and/or the antimalarial rescue medications and the event.  
In addition to the assessments of relationship to the trial intervention/s, the Investigator should comment 
on the AE record in the eCRF whether an AE is related to the trial participation of the participant (trial 
procedures etc.).  
Of note, a sign or symptom associated with malaria infection (confirmed by a positive malaria 18S qPCR 
at the onset of the event) that is of expected intensity, frequency and duration for the individual 
participant in the context of this trial is considered to be a malaria challenge agent-related event.  
8.5.4.3 EXPECTEDNESS  
A ‘Suspected Unexpected Serious Adverse Reaction (SUSAR)’ is a Serious AR that is unexpected. The 
‘expectedness’ of a serious AR is assessed in the light of the Reference Safety Information (RSI) of the 
investigator brochures. 
8.5.5 TIME PERIOD AND FREQUENCY FOR EVENT ASSESSMENT AND FOLLOW-UP 
All AEs must be documented and followed up by an Investigator until: 
 
the event is resolved, or 
 
no further medically relevant information in relation to the event can be expected, and 
 
an Investigator considers it justifiable to terminate the follow-up 
Events that are unresolved at the time of the participant’s last follow-up visit should continue to be 
followed up by an Investigator for as long as medically indicated or until the participant is referred to a 
general practitioner or medical specialist as appropriate. The Sponsor retains the right to request 
additional information for any participant with ongoing AE(s)/SAE(s) at the end of the trial, if judged 
necessary. 
All AEs should be treated appropriately. An Investigator will decide upon the appropriate action to be 
taken in response to an AE, which may include one or more of the following: 
 
no action taken (i.e., further observation only), or 
 
administration of a concomitant medication, or 
 
hospitalisation or prolongation of current hospitalisation (event to be reported as an SAE), or 
 
other 
In a case of occurrence of an SAE, regardless of whether or not it is judged to be related to any of the trial 
treatments or procedures, the participant will receive appropriate care under clinical supervision until all 
the symptoms of the SAE have diminished or resolved and the participant’s condition has improved. 


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60 
For ongoing AEs, care will be provided for a period of time as specified in the clinical trial unit work 
instruction protocols. However, if the nature of the ongoing AE is determined by an Investigator as not 
being associated with any of the trial treatments or procedures, the participant will be advised to visit 
his/her general practitioner for further clinical care that might be required. 
8.5.6 ADVERSE EVENT REPORTING  
It is Investigator’s responsibility to document and report all AEs occurring in the clinical trial whether 
spontaneously reported by the participant, observed by an Investigator (either directly or by laboratory 
or other assessments), or elicited by general questioning. The period of observation for collection of AEs 
extends from the time of inoculation with the malaria challenge agent to EOS. These AEs must be recorded 
on specific AE pages of the eCRF. Events reported prior to this will be recorded as medical history, unless 
the symptoms worsen during the trial. 
The following information should be recorded for all AEs: 
 
Description 
 
Dates and times of onset and resolution 
 
Duration in hours 
 
Time of onset relative to IMP administration, inoculation with the malaria challenge agent, and/or 
administration of antimalarial rescue medications 
 
Seriousness (SAE or not) 
 
Severity 
o If severity of an AE changes, only one AE will be reported, with the highest severity 
recorded in the eCRF and listings and tables. In the clinical file (source data), the 
description of the AE will report the various severities observed over time. 
o If the AE resolves and then reoccurs, then two AEs will be reported. 
 
Action taken in response to the AE (including treatment given). 
 
Outcome. 
 
Relationship to the trial treatments or procedures (causality assessment), including malaria challenge 
agent, antimalarial medication, or any other treatment or procedure conducted during the trial.  
8.5.7 SERIOUS ADVERSE EVENT AND EVENT OF SPECIAL INTEREST REPORTING  
An Investigator will take immediate appropriate action in response to SAEs and AESIs to ensure participant 
safety and in an attempt to identify the cause of the event. The Investigator must notify the SAE or AESI 
to the pharmacovigilance provider (Prime Vigilance) by email within 24 hours of becoming aware of the 
event.  
All reports must be signed by the Principal Investigator or delegate and notified to Prime Vigilance 
preferably by email or fax to: 
Email: MMV@primevigilance.com 
Back-up fax number: +44 800 471 5694 


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61 
Prime Vigilance Contact:  
 
Head Office: +44 1483 307920 
Any copies of participant’s medical records provided for SAE or AESI reporting must have all participant 
identifiers redacted before submission.   
The report forms will always be completed as thoroughly as possible with all available details of the event 
and signed by the Principal Investigator or delegate. If the Principal Investigator or delegate does not have 
all information regarding an event, he/she will not wait to receive additional information before reporting 
the event. A follow-up SAE or AESI report should be completed within 14 days, or if there is no new 
information the report form should be updated when additional information is received. 
The Principal Investigator or delegate will always provide an assessment of causality at the time of the 
initial report. 
Email transmission of the report forms is the preferred method to transmit this information to Prime 
Vigilance. In rare circumstances notification by telephone is acceptable, with a copy of the report form 
sent by overnight mail.  
The Principal Investigator or delegate, or responsible person according to local requirements, will comply 
with the applicable local regulatory requirements related to the reporting of SAEs to regulatory authorities 
and the HREC. Any SAE that meets the criteria of a SUSAR will be reported to the TGA by the Sponsor in 
accordance with the Sponsor’s reporting procedures. 
8.5.8 REPORTING EVENTS TO PARTICIPANTS    
 
Not applicable 
 
8.5.9 REPORTING OF PREGNANCY  
Pregnancy itself is not defined as an AE/SAE. Any complication or termination of pregnancy for medical 
reasons are to be reported as an AE/SAE. Spontaneous abortion, still birth or congenital anomaly must be 
reported 
as 
an 
SAE. 
Any  
WOCBP 
enrolled  
in  
the  
study 
who  becomes  pregnant  during  the  study  and  the  following 100 days after pyronaridine administration 
should be followed through delivery or termination of the pregnancy. The Investigator will collect 
pregnancy information and report to Primevigilance within 24 hours of becoming aware of a participant’s 
pregnancy using the pregnancy form and following the same process as described in section 8.5.7 for SAEs 
and AESIs. Pregnancy follow-up should be recorded on the same form including pregnancy outcomes. 
Once pregnancy is confirmed, pregnant female participants will be immediately withdrawn from the study 
and will be managed as per section 7.2. Where possible, the Investigator will also attempt to collect and 
report information regarding pregnancy outcomes of female partners of any male participants who were 
administered pyronaridine in this study and the following 70 days after pyronaridine administration. 
Appropriate 
signed 
informed 
consent 
will 
be 
required  directly  from  the  pregnant  female  partner  to  obtain  and  report  this  information.    Any 


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62 
participant’s female partner who becomes pregnant during the study should be followed through delivery 
or termination of the pregnancy.  
9 
STATISTICAL CONSIDERATIONS  
The following sections describe the statistical analysis as it is foreseen when the study is being planned. A 
detailed Statistical Analysis Plan (SAP) and Modelling Analysis Plan (MAP) will be finalised and approved 
prior to database lock. The SAP and MAP will provide details of all analyses to be performed as well as the 
format of listings and tables to be provided for completion of the Clinical Study Report (CSR). Any 
deviations from the SAP will be described and justified in the final CSR. 
9.1 
STATISTICAL HYPOTHESES 
The primary objective of this study is to characterise the PK/PD relationship of pyronaridine in healthy 
participants experimentally infected with blood stage P. falciparum. There is no pre-specified statistical 
hypothesis for testing this objective. 
9.2 
SAMPLE SIZE DETERMINATION 
A maximum sample size of 18 participants is expected to be sufficient to achieve the primary endpoint of 
the study, which is to determine the PK/PD parameters of pyronaridine. This sample size is not based on 
formal statistical calculations but rather is based on previous experience of characterising the PK/PD 
relationship of antimalarials using the IBSM study. The specific number of participants required will be 
dependent on the emerging results during the study; it is possible that less than 18 participants will be 
required. The SDRT will review the data after the completion of each cohort and decide when sufficient 
data has been obtained to complete the primary endpoint. No more than 18 participants will be enrolled 
in this study. 
9.3 
POPULATIONS FOR ANALYSES 
In the first instance, two (2) analysis datasets will be used for study analyses: Full Analysis Set (FAS) and 
Safety Set. Additional analysis populations like the ones to be used for calculation of PK/PD parameters, 
will be defined in the SAP. The number of participants in each analysis set will be summarised, with a 
corresponding listing.  
The FAS will consist of all enrolled participants. The FAS will be used to assess all participant disposition, 
baseline, demographic and protocol deviation data. The safety analysis dataset will include all participants 
who receive the malaria challenge agent on Day 0.  
9.4 
STATISTICAL ANALYSES 
9.4.1 GENERAL APPROACH 


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Continuous variables will be summarised with the number of observations (non-missing values), mean, 
standard deviation, median, quartiles, minimum and maximum. The minimum and maximum values will 
be presented to the same number of decimal places as recorded in the raw data. The mean, median and 
SD will be presented to one more decimal place than the raw data. Categorical variables will be 
summarised with the number of observations and the numbers and percent from each category. 
Percentages will be rounded to one decimal place, with the denominator being the number of participants 
in the relevant population with non-missing data. No missing data will be imputed. If required, CIs will be 
two sided and will use 95% confidence levels. Any analyses requiring significance testing will use a two-
sided test at the 5% significance level. 
The inoculation baseline will be defined as the last available assessment prior to the inoculation. The IMP 
baseline will be defined as the last available assessment prior to the administration of the IMP. 
Unscheduled visits will be excluded from summary tables. Assessments conducted at Early Termination 
will be excluded from summary tables. 
9.4.2 ANALYSIS OF THE PRIMARY ENDPOINT 
The primary endpoint is the PK/PD relationship of pyronaridine. The basic PK/PD relationship will link the 
drug killing rate to its blood concentration with a sigmoidal Emax model characterized by the Emax, EC50 and 
Hill coefficient parameters. If needed alternative models will be applied, for instance to account for a lag 
in the drug response, which will contain additional parameters. The parameters will be estimated using a 
non-linear mixed-effect modelling of pyronaridine concentration-time and parasitaemia-time profiles. 
First, a population PK model will be constructed with the measured concentrations; it will allow to obtain 
the individual PK parameters and calculate the full PK profile for each individual. Second, the PD 
parameters will be estimated with the measured parasitaemia and the predicted individual PK profiles. 
As a general concept the changes in living parasite 𝑃 are modelled as the effect of a net exponential 
growth rate 𝐺𝑅 and a killing or clearance rate 𝐾𝑖𝑙𝑙 due to pyronaridine with an initial parasitaemia 𝑃𝑏𝑎𝑠𝑒 
at time 𝑡0 (time of first observation). The equations are expressed in the log-scale such as:  
𝑑𝑃𝐿
𝑑𝑡
=
𝐺𝑅−𝐾𝑖𝑙𝑙
𝑃𝐿(𝑡0)
=
𝑃𝐿𝑏𝑎𝑠𝑒
 
 where 𝑃𝐿= ln(𝑃) is the log-transformed parasite counts. 
The 
effect 
of 
concentrations 
on 
parasite 
killing/clearance 
will 
be 
described 
as: 
𝐾𝑖𝑙𝑙= 𝐸𝑚𝑎𝑥⋅
𝐶𝑐𝐻𝑖𝑙𝑙
𝐶𝑐
𝐻𝑖𝑙𝑙+ 𝐸𝐶50
𝐻𝑖𝑙𝑙 
where 𝐶𝑐 is the concentration in the central compartment, 𝐸𝑚𝑎𝑥 is the maximum effect of the drug, 𝐸𝐶50 
is the concentration that results in 50% of the maximum effect and 𝐻𝑖𝑙𝑙 is the Hill coefficient. 
Alternative models may be tested if the exploration of the data indicate a different behaviour, e.g. if a lag 
time is observed before the parasites are cleared.  


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Other key parameters such as the minimum inhibitory concentration (MIC), the minimal parasiticidal 
concentration (MPC90), and the parasite reduction rate in 48 h (log10PRR48) will be derived from the PD 
model. The MIC is defined as the concentration when parasite clearance by the drug equals the parasite 
growth, i.e., the time at which the minimum parasite concentration is observed. It is calculated with the 
following equation in case of the Emax model. 
𝑀𝐼𝐶= 𝐸𝐶50 (
𝐺𝑅
𝐸𝑚𝑎𝑥+ 𝐺𝑅)
1/𝐻𝑖𝑙𝑙
 
The MPC90 is defined as the concentration at which the clearance effect is at 90% of the maximum. It is 
calculated as follows: 
𝑀𝑃𝐶90 = 𝐸𝐶50 ⋅91/𝐻𝑖𝑙𝑙 
The PRR48 is defined as the parasite clearance achieved within 48 hours, usually given as the reduction of 
values on log10 transformed scale. The maximum capacity of parasite clearance in 48 hours is determined 
as follows assuming that concentrations are maintained well above the MPC90 for this time span. 
𝑃𝑅𝑅48 = 48ℎ𝐸𝑚𝑎𝑥−𝐺𝑅
ln(10)
 
Full details of the modelling analysis will be described in the SAP. 
PK and PD parameters will be tabulated as population estimates and relative standard error (RSE).  
9.4.3 ANALYSIS OF THE SECONDARY ENDPOINTS 
This study has three secondary endpoints.   
Safety parameters: 
The safety and tolerability of pyronaridine will be assessed by clinical review of the following parameters: 
       AEs (including SAEs and AESIs) 
       Vital signs 
       12-lead ECG 
       Haematology, Biochemistry, urinalysis 
       Physical examination 
The analysis of this endpoint is discussed in Section 9.4.4 below.  
Parasite clearance kinetics: 
The parasite clearance kinetics following dosing with of pyronaridine will be determined by calculating 
the parasite reduction ratio over a 48 hour period (PRR48) and corresponding parasite clearance half-life 
(PCt1/2); the incidence of parasite regrowth (n and % per dose group) will be recorded. 


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The PRR48 will be estimated using the slope of the optimal fit of the log-linear relationship of the 
parasitaemia decay. The optimal fit will be derived using summarised replicate parasitaemia data, which 
have been cleaned by dealing with potential outliers, values below the limit of detection and non-
detectable values (ND). The optimal fit of the log-linear parasitaemia-by-time relationship is determined 
by using left and right censoring to systematically remove the potential lag phase and tail phase of the 
parasitaemia decay. The decay rate, estimated by the slope coefficient from the log-linear decay 
regression, will be calculated for each participant. The overall dose-specific PRR48 will be estimated with 
its 95% CI by calculating the weighted average slope estimate and corresponding standard error (SE) using 
an inverse-variance method. Only participants who have optimal regression models with appropriate fit 
contribute toward the dose-specific PRR48. Details will be presented in the SAP. 
The parasite clearance half-life (Pt ½) will be derived from the optimal decay rate. PCt1/2 is a 
transformation of PRR48 into a per hour clearance rate. Details regarding the calculation of this parameter 
will be presented in the SAP.    
PK parameters: 
The following PK parameters will be determined using non-compartmental methods from whole blood 
pyronaridine concentration-time profiles: AUClast, AUCinf, Cmax, tmax, t1/2, CL/F, Vd/F and λ. All PK variables 
(normally and log-normally distributed) will be summarized using arithmetic and geometric means, 
minimum, median, maximum, SD and coefficient of variation (CV %) of arithmetic and geometric means. 
Summary tables showing the median and range for tmax and geometric mean and CV% for all other PK 
variables will be provided by day and dose. 
 Whole blood concentration, values below the lower limit of quantification (LLOQ) will be set to zero. All 
mentioned parameters will be expressed as mean ± SD, unless otherwise stated. Details regarding the 
calculation of this parameter will be presented in the SAP.    
9.4.4 SAFETY ANALYSES 
 
All descriptive statistics for safety parameters will be evaluated using the Safety Set.  
All AE data will be listed for each volunteer, including severity, relationship to IMP (pyronaridine and 
malaria challenge agent), relationship to non-IMP protocol-specific treatments, outcome and actions 
taken. In addition, listings of AEs leading to discontinuation of the study, AESIs, SAEs and deaths, will be 
provided as applicable. Adverse events (AE) will be coded according to MedDRA and grouped by SOC and 
PT. 
The main AE summaries will be restricted to Treatment Emergent Adverse Events (TEAEs) only, where a 
TEAE is defined as an AE that commences on, or after, the first administration of IMP up to the end-of-
study (EOS) visit (inclusive). TEAEs without an onset date or time will be defined as treatment emergent 
except if an incomplete date (e.g., month and year) clearly indicates that the event started prior to first 
administration of IMP, or if the AE stop date indicates that the event stopped prior to the first 
administration of IMP. 


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At a minimum, the following AE summary tables will be provided:  
• Overall summary of AEs  
• All TEAEs  
• TEAEs by severity  
• TEAEs by relationship to the IMP  
• TEAEs by relationship to rescue medications  
• Serious TEAEs  
• TEAEs leading to study withdrawal  
• AESIs  
• All Inoculation Period AEs  
• Inoculation Period AEs by relationship to the malaria challenge agent  
Lab parameters, physical examination, Body measurements, vital signs, Beck Depression inventory score 
and 12-lead ECG will be listed for all participants and visits. Summary tables for each scheduled visit, 
including observed values, change from each baseline and clinical significance of each abnormal value, 
will be presented.  
9.4.5 BASELINE DESCRIPTIVE STATISTICS 
 
Demographic data will be summarised by descriptive statistics and listed for all enrolled participants.  This 
will include total number of observations, mean, standard deviation, and range for continuous variables 
that are normally distributed for medians and interquartile ranges for non-parametric continuous 
variables, and number and percentages for categorical variables.  
Volunteer disposition will be summarised and presented in a flow diagram. Trial completion, trial 
withdrawals, exclusions, and violations will be summarised and the reasons for withdrawal, exclusions, 
and violations will be listed.  
Medical history, current medical conditions, prior and concomitant medications, results of laboratory 
screening tests, drug and alcohol screening tests, and any other relevant baseline information will be listed 
by participant and treatment group.  Medical history will be coded using MedDRA and summarised by 
SOC and PT. 
9.4.6 PLANNED INTERIM ANALYSES  
 
This study has no formal interim analyses other than review of safety, tolerability, parasitaemia and PK 
parameters by the SDRT between cohorts. 
 
9.4.7 SUB-GROUP ANALYSES 


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Not applicable. 
 
9.4.8 TABULATION OF INDIVIDUAL PARTICIPANT DATA 
 
All individual participant data will be listed by measure and time-point. 
 
9.4.9 EXPLORATORY ANALYSES 
 
Not applicable. 
 
10 SUPPORTING DOCUMENTATION AND OPERATIONAL CONSIDERATIONS 
 
10.1 REGULATORY, ETHICAL, AND STUDY OVERSIGHT CONSIDERATIONS 
10.1.1 ETHICAL STANDARD 
The trial will be conducted in accordance with the protocol approved by the HREC(s), the principles of the 
Declaration of Helsinki (Recommendations guiding Medical Doctors in Biomedical Research Involving 
Human Participants, Fortaleza, Brazil 2013), the NHMRC National Statement on Ethical Conduct in Human 
Research (2007, updated 2018) and the Integrated Addendum to ICH E6 (R1): Guideline for Good Clinical 
Practice E6 (R2) (November 2016) — with introductory comments of the Australian TGA. 
The PI will minimise any discomfort experienced by participants during the trial. The only invasive 
procedures will be the IV inoculation of the malaria challenge agent and the blood collection by 
cannulation/venepuncture. The total volume of blood drawn from each participant will not exceed 470 
mL in any 30-day period during trial participation. 
10.1.2 ETHICAL REVIEW 
The protocol, Participant Information Sheets, and Informed Consent Forms will be reviewed by the 
approving HREC(s) and no trial activities will be initiated prior to approval. All amendments and addenda 
to the protocol and consent forms will similarly be submitted to the approving HREC(s) for approval prior 
to their implementation. 
Changes to the final trial protocol can only be made with the prior consent of the PI, the Sponsor and the 
approving HREC(s). All such changes must be attached to (or incorporated into) the final protocol and 
communicated to all relevant members of the clinical trial unit staff and, if appropriate, to trial 
participants. All deviations from this trial protocol will be included in the trial master file and included in 
the CSR. The types of amendments are discussed below. 
 
 


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Administrative or minor changes  
Administrative or minor changes include but are not limited to changes in trial staff or contact details (e.g., 
Sponsor instead of contract research organisation monitors). Amendments for administrative or minor 
changes may be suitable for executive review (expedited) by the approving HREC(s). 
Substantial amendment 
Significant changes require a substantial amendment. Significant changes include but are not limited to: 
new data affecting the safety of participants, change of the objectives/endpoints of the trial, eligibility 
criteria, dose regimen, trial assessments or procedures, treatment or trial duration with or without the 
need to modify the Participant Information Sheet and Informed Consent Form. Substantial amendments 
are to be approved by the HREC(s). The implementation of a substantial amendment can only occur after 
formal approval from the Sponsor, HREC(s), and PI. 
Urgent amendment 
An urgent amendment might become necessary to preserve the safety of the participants included in the 
trial. The requirements for approval should in no way prevent any immediate action being taken by the PI 
or the Sponsor in the best interests of the participants. Therefore, if deemed necessary, the Investigator 
can implement an immediate change to the protocol for safety reasons with notification to the medical 
monitor as soon as practicably possible. This means that, exceptionally, the implementation of urgent 
amendments will occur before submission to, and approval by, the HREC(s). In such cases, the PI must 
notify the Sponsor within 24 hours. A related substantial amendment will be written and submitted to 
HREC(s) as soon as practicable but no later than 7 working days, together with a description of the steps 
that have already been taken in regard to implementation of this amendment. 
HREC approval of future research 
In the event that the PI or the Sponsor want to perform testing on the samples that is not described in the 
protocol, additional approval will be sought from the approving HREC(s). This may be done if a participant 
consents to blood storage for use in future research.  
10.1.3 INFORMED CONSENT PROCESS 
 
10.1.3.1 CONSENT/ASSENT AND OTHER INFORMATIONAL DOCUMENTS PROVIDED TO 
PARTICIPANTS 
Each potential participant will be given the trial Participant Information Sheet and Informed Consent Form 
that describes in detail the trial interventions, trial procedures, and risks.  
All participants will receive an Informed Consent for Malaria Future Research and an option to grant 
permission to be contacted about involvement in future trials. If a potential participant chooses not to 
consent to these options this will not impact their eligibility for the trial.   
 


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10.1.3.2 CONSENT PROCEDURES AND DOCUMENTATION 
During the initial screening visit/recruitment, potential participants will read the Participant Information 
Sheet. An Investigator will explain the trial via the Participant Information Sheet and the candidate 
participants will be encouraged to ask questions. Potential participants will be informed that participation 
is voluntary and that they may withdraw from the trial at any time, without prejudice. Potential 
participants will have the opportunity to discuss the trial with their family or think about it prior to 
agreeing to participate.  
Individuals willing to be considered for inclusion in the trial will sign and date the Informed Consent Form 
in the presence of an Investigator. Participants will be given a copy of their signed Informed Consent Form. 
The conduct of the informed consent process will be documented in the source document (including the 
date) and the form will be signed before the participant undergoes any trial-specific procedures; only once 
the participant has consented to the trial may trial-specific screening activities commence. 
 
10.1.4 STUDY DISCONTINUATION AND CLOSURE 
The Sponsor, PI, approving HREC(s), and regulatory authorities independently reserve the right to 
discontinue the trial at any time for safety or other reasons. Circumstances that may warrant termination 
or suspension include, but are not limited to: 
 
determination of unexpected, significant, or unacceptable risk to participants 
 
insufficient compliance with protocol requirements 
 
data that are not sufficiently complete and/or evaluable 
 
determination that the primary endpoint has been met 
This will be done in consultation with the Sponsor where practical. In the event of premature trial 
termination or suspension, the above-mentioned parties will be notified in writing by the 
terminator/suspender stating the reasons for early termination or suspension (with the exception of the 
Sponsor’s responsibility for notifying the regulatory authorities). After such a decision, the Sponsor and 
the Investigator will ensure that adequate consideration is given to the protection of the participants’ 
interest and safety. The Investigator must review all participants as soon as practical and complete all 
required records. 
10.1.5 CONFIDENTIALITY AND PRIVACY  
Participants will be informed that their data will be held on file by the clinical trial unit and that these data 
may be viewed by staff of the clinical trial unit (including, where necessary, staff of the clinical trial unit 
other than the named Investigators).  
In the event of a notifiable disease being discovered during the trial, the appropriate Public Health 
authorities will be notified in accordance with the Queensland Public Health Regulation 2018.  


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Upon request, the Investigator(s)/institution(s) will permit direct access to source data and documents 
for trial-related monitoring, audits, HREC review, and regulatory inspection(s) by the Sponsor (or their 
appropriately qualified delegates) and regulatory authorities.  
Participants will also be informed that a report (CSR) of the trial will be submitted to the Sponsor and may 
also be submitted to regulatory authorities and perhaps for publication, but that they will only be 
identified in such reports by their trial identification number, their sex and age. The Investigators will 
undertake to hold all personal information in confidence. 
Participants will be informed that samples collected for the purposes described in the protocol will be 
sent to the Sponsor’s nominated national or international laboratory for assessment. 
10.1.6 FUTURE USE OF STORED SPECIMENS AND DATA  
As part of the trial, safety serum retention samples will be stored for 15 years at QIMR Berghofer for 
retrospective safety assessments that may later be indicated. Participants consent to this storage and the 
use of the sample for safety assessments when they sign the Informed Consent Form for the trial. 
For all other samples, consent must be obtained from the participants to store and use their samples for 
malaria future research.  
Under a separate protocol and consenting process, blood samples may be collected for storage and use 
in future malaria research. The separate study will be under the local sponsorship of QIMR Berghofer 
Institute and will involve a protocol and consent document approved by the QIMR HREC. All samples will 
be stored at QIMR Berghofer in accordance with the laboratory SOPs. The Investigator will ensure that 
confidentiality will be maintained continuously in all future research that involves use of these samples. 
No genetic testing will be performed on the stored samples without obtaining consent from the 
participants. The stored samples will not be sold or used directly for production of any commercial 
product. There are no benefits to participants in the collection, storage, and subsequent research use of 
their samples. Reports about future research done with participant samples will NOT be kept in participant 
health records, but a participant’s samples may be kept with the trial records or in other secure areas.  
 
10.1.7 KEY ROLES AND STUDY GOVERNANCE 
 
Principal Investigator 
A/Prof. Bridget Barber MBBS, DTM&H, MPH, FRACP, PhD 
QIMR Berghofer Medical Research Institute 
Level 5, 300C Herston Road 
Herston QLD 4006, Australia 
Tel: +61 424 737 153 
Email: Bridget.Barber@qimrberghofer.edu.au 
Sponsor Medical Director 
Anne Claire Marrast, MD  
Medicines for Malaria Venture  
Route de Pré-Bois 20  
1215 Geneva 15  


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71 
Switzerland  
Email: marrasta@mmv.org 
Sponsor Project Director  
 
 
 
 
 
Jörg Möhrle, PhD  
Medicines for Malaria Venture  
Route de Prés-Bois 20  
1215 Geneva 15  
Switzerland  
Tel: +41 22 555 0330  
Email: moehrlej@mmv.org  
Local Study Sponsor  
Southern Star Research Pty Ltd (SSR) 
1 Merriwa St 
Gordon, NSW 2072 
Australia 
Tel: +61 (0)2 9011 6266 
Study Monitor   
Southern Star Research Pty Ltd (SSR) 
1 Merriwa St 
Gordon, NSW 2072 
Australia 
Tel: +61 (0)2 9011 6266 
Institutional Ethics Committee 
QIMR Berghofer Medical Research Institute Human Research Ethics 
Committee (QIMR Berghofer HREC; EC00278) 
Locked Bag 2000, Royal Brisbane and Women’s Hospital, Brisbane 
QLD 4029, Australia 
Tel: +61 (0)7 3362 0117 
Biostatistician (parasitaemia) 
Stacey Llewellyn 
QIMR Berghofer Medical Research Institute 
300C Herston Rd 
Herston QLD 4006, Australia 
Tel: +61 (0)7 3845 3579 
Email: Stacey.Llewellyn@qimrberghofer.edu.au 
Biostatistician 
Southern Star Research Pty Ltd (SSR) 
1 Merriwa St 
Gordon, NSW 2072 
Australia 
Tel: +61 (0)2 9011 6266 
Pharmacometrician 
Nathalie Gobeau 
Medicines for Malaria Venture  
Route de Pré-Bois 20  
1215 Geneva 15  
Switzerland  
Email: gobeaun@mmv.org 
Independent Medical Monitor  
Dr Joanne Marjason 
Southern Star Research Pty Ltd 
1 Merriwa St 
Gordon, NSW 2072, Australia 
Tel: +61 (0)427 959 156 
Email: jmarjason@southernstarresearch.com 


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Clinical Study Centre 
USC Clinical Trials Unit 
 
Moreton Bay  
Health Hub Morayfield  
19-31 Dickson Road Morayfield QLD 4506 Australia 
Tel: +61 (0)7 54563965 
Email: ctchhm@usc.edu.au 
 
South Bank 
Building A2, SW1 Complex 
52 Merivale Street South Brisbane QLD 4101 
Tel: +61 (0)7 5409 8630 
Email: ctcsouthbank@usc.edu.au 
Laboratories 
Clinical laboratory measurements: 
Sullivan Nicolaides Pathology Central Laboratory (SNP) 
24 Hurworth Street  
Bowen Hills, QLD  4006, Australia 
Tel: +61 (0)7 3377 8782 
 
Parasite quantification in blood samples: 
Ms. Claire Wang 
Queensland Paediatric Infectious Diseases Laboratory  
(Q-PID), SASVRC, Level 8, Centre for Children’s Health Research 
62 Graham Street 
South Brisbane, QLD 4101, Australia 
Tel: +61 (0)7 3069 7462 
Email: claire.wang@uq.edu.au 
 
Investigational drug quantification in blood samples: 
Dr. Christoph Siethoff CEO 
Swiss BioQuant AG,  
Kägenstrasse 18 
4153 Reinach, Switzerland 
Tel:  +41 (0) 617 1698 12         
Email: Christoph.Siethoff@swissbioquant.com, 
mail@swissbioquant.com 
Serious AE and AESI reporting 
Prime vigilance 
Tel: +44 (0) 800 471 5694  
Email: mmv@primevigilance.com 
 
10.1.8 SAFETY OVERSIGHT 
The SDRT will be responsible for decisions related to the safety of participants and the continuation of the 
trial. The SDRT will meet to assess any event/s that trigger the discontinuation rules or as needed to 
provide a recommendation and findings to the approving HREC(s) and the PI. 


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The SDRT will meet at the end of each cohort to review safety data (AEs, AESIs, SAEs, and laboratory data), 
parasitaemia data, and the available PK and PK/PD modelling data.  Based on this review, the SDRT will 
decide on whether to progress to the next cohort and will decide on the doses of pyronaridine to be 
administered.  
The SDRT may also decide to modify the frequency and timing of blood sample collection for malaria 18S 
PCR and/or pyronaridine blood concentration measurements as long as blood volume is not greater than 
470 mL in any 30-day period  
The role and composition of the SDRT is outlined in the study specific SDRT Charter, but at a minimum the 
SDRT will be comprised of; 
 
MMV Medical Director or delegate  
 
MMV Vice President, Head of Translational Medicine or delegate 
 
Principal Investigator (PI) or delegate or delegate 
 
Local Sponsor Medical Monitor or delegate 
 
MMV Pharmacometrician or delegate (ad hoc as required) 
 
10.1.9 CLINICAL MONITORING 
It will be the Sponsor’s responsibility to ensure that the trial is monitored in accordance with the 
requirements of GCP. The conduct of the trial will be reviewed internally by the clinical trial unit in 
accordance with their SOPs and work instructions, and GCP guidelines. The trial will be monitored 
according to the SOPs of the Sponsor and all serious breaches, suspected breaches and protocol deviations 
will be reported to the Sponsor. Serious breaches that impact participant safety or data integrity will also 
be reported to the approving HREC. 
During the trial, trial monitor(s) (on behalf of the Sponsor) will visit the clinical trial unit regularly (or, 
under exceptional circumstances, access remotely the clinical unit files) to check the completeness of 
participant records, accuracy of eCRF entries, adherence to the protocol and to GCP, the progress of 
enrolment, and to ensure that trial interventions are being stored, dispensed, and accounted for according 
to specifications. Key trial personnel will be required to be available to assist the monitor during these 
visits (or provide the Monitor with an access to the required documents, if the visit is performed 
remotely).  
The Investigator will be required to give the monitor access to all relevant source documents to confirm 
their consistency with the eCRF entries. At a minimum, the Sponsor will require full verification for the 
presence of informed consent, adherence to the inclusion/exclusion criteria, and documentation of SAEs, 
AESI and AEs, and the recording of data that was used for all primary and safety variables. Additional 
checks of the consistency of the source data with the eCRFs will be performed according to the trial-
specific monitoring plan. No information captured in the source documents about the identity of the 
participants will be disclosed. 
 


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10.1.10 
QUALITY ASSURANCE AND QUALITY CONTROL 
The clinical trial unit will perform internal quality management of trial conduct, data and biological 
specimen collection, documentation and completion.   
Quality control procedures will be implemented beginning with the data entry system and data quality 
control checks that will be run on the database that will be generated. Any missing data or data anomalies 
will be communicated to the site for clarification and resolution. 
The clinical trial unit will provide direct access to source data and documents, and reports for the purpose 
of monitoring and auditing by the Sponsor, and inspection by regulatory authorities. 
Data management, including the development and management of a secure database, will be performed 
in accordance with regulatory requirements. The designated data management vendor will review the 
data entered into the eCRFs by clinical trial unit staff for completeness and accuracy. A formal querying 
process will be followed whereby the data management team will request the site personnel to clarify 
any apparent erroneous entries or inconsistencies and will request additional information from the site 
as required. 
Medical history/current medical conditions and AEs will be coded using the Medical Dictionary for 
Regulatory Activities (MedDRA) terminology (version 20.1 or higher). Prior and concomitant medications 
will be coded using the WHO Drug Dictionary Enhanced (WHO Global; March 2014 or later). 
 
After all data have been captured and reviewed, all queries have been resolved with the site, and any 
protocol deviations that were identified during the data management processes have been confirmed by 
the site, the database will be declared to be complete and accurate. The database will be locked and made 
available for data analysis. Any changes to the database after that time may only be made, in consultation 
with the Sponsor and in accordance with documented database unlock and relock procedures. 
 
10.1.11 
DATA HANDLING AND RECORD KEEPING  
 
10.1.11.1 
DATA COLLECTION AND MANAGEMENT RESPONSIBILITIES  
Participants will be assigned a unique identifier when participating in the study, and any participant 
datasets or records that are transferred to the Sponsor or CRO will only contain this identifier. Any other 
identifiable information about the participant will not be transferred.  The PI will ensure procedures are 
in place to appropriately protect the confidentiality of the participant records and data, including 
adequate safe guards for digital/computer access. The participants will be informed that their personal 
study-related data will be used by the Sponsor and that their medical records may be examined by 
auditors and regulatory agencies.   
Each participant will have a clinical file (source data) and eCRF (for protocol specific data) into which 
relevant data will be recorded. All recording of source data and documents will be done in RealTime, the 


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75 
clinical trial unit’s eSource system. Pathology reports, ECGs and any other paper source will be uploaded 
to RealTime.  
Guidelines for eCRF completion including correcting data and responding to data queries will be provided 
by the Sponsor or CRO. A log of names, signatures, and initials of all staff authorised to enter data into a 
participant’s clinical file and eCRF will be kept. Corrections to source document (whether paper or 
electronic) must have an audit trail (i.e., must not obscure or delete the original entry, and the date/time 
of correction and identity of the person making the correction must be clearly indicated).  
10.1.11.2 
STUDY RECORDS 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. 
10.1.12 
PROTOCOL DEVIATIONS  
Protocol deviation: any departure, change, and/or addition from the trial design or procedures defined in 
the protocol that has received approval by the competent authorities and favourable opinion from the 
approving HREC.  
Suspected serious breach: a report that is judged by the reporter as a possible serious breach, but has yet 
to be formally confirmed as a serious breach by the Sponsor. 
Serious breach: a breach of GCP or the protocol that is likely to affect to a significant degree: a) the safety 
or rights of a trial participant, and/or b) the reliability and robustness of the data generated in the clinical 
trial. 
Note to File (NTF): a record that documents in detail actions taken, important decisions made, or explains 
a sequence of events where no other detailed record exists to enable the conduct of the trial to be 
reconstructed. 


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Reporting requirements: 
 
All protocol deviations, suspected serious breaches, and serious breaches will be documented in 
the trial master file and included in the CSR. 
 
All NTFs, protocol deviations, suspected serious breaches, and serious breaches will be viewed by 
the PI’s delegate and signed by the PI. 
 
All protocol deviations, suspected serious breaches and serious breaches will be reported and 
assessed at each SDRT meeting. 
 
All serious breaches will be reported by the clinical trial unit to the Sponsor and the approving 
HREC(s) as early as possible, but within 7 days.  
 
All participant-specific protocol deviations will be captured in a log by the clinical site and reported 
by the clinical trial unit to the PI’s delegate and signed by the PI at the end of each cohort and 
shared with the Sponsor for review and sign off before database lock. 
 
Protocol deviation logs will be submitted by the clinical trial unit to the Sponsor and approving 
HREC via inclusion with the annual report.  
10.1.13 
PUBLICATION AND DATA SHARING POLICY 
The data management, statistical, and medical writing team appointed by the Sponsor will collaborate to 
provide a detailed CSR upon conclusion of the trial. The CSR will include appendices of all tables and 
listings generated during the analyses of data. The tables, figures and listings will be provided by the 
Sponsor. The Sponsor will undertake to ensure that all safety observations made during the conduct of 
the trial are documented in the CSR. 
Publication and reporting of results and outcomes of this trial will be accurate and honest, and undertaken 
with integrity and transparency. The Sponsor recognises that the PI has a responsibility to ensure that 
results of scientific interest arising from the trial are appropriately published and disseminated. 
Publication of results will be volunteered to fair peer-review. Authorship will be given to all persons 
providing significant input into the conception, design, and execution or reporting of the research. No 
person who is an author, consistent with this definition, will be excluded as an author without his/her 
permission in writing. Authorship will be discussed between researchers prior to trial commencement (or 
as soon as possible thereafter) and reviewed whenever there are changes in participation. 
Acknowledgement will be given to collaborating institutions and hospitals and other individuals and 
organisations providing finance or facilities.  
Neither the complete nor partial results of the study achieved under this protocol, nor any of the 
information provided by the Sponsor for the purposes of performing the study, will be published or passed 
on to any third party without the consent of the study Sponsor.  Any Investigator involved with this study 
is obligated to provide the Sponsor with complete study results and all data derived from the study. 
The Sponsor will ensure that the key design elements of this protocol are posted in a publicly accessible 
database such as Australian New Zealand Clinical Trials Registry (ANZCTR) or Clinicaltrials.gov. In addition, 
upon trial completion and finalisation of the CSR, the results of this trial will be submitted for publication 
in an open access journal and/or posted in a publicly accessible database of clinical trial results.  


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10.1.14 
CONFLICT OF INTEREST POLICY 
Potential conflicts of interest will be identified, assessed, managed and documented, with any action 
taken and reasons for taking action recorded during the ongoing risk management process. 
 
10.2 LIABILITY/INDEMNITY/INSURANCE 
  
The Local Sponsor will ensure sufficient insurance is available to enable it to indemnify and hold the 
Investigators and relevant staff as well as any hospital, institution, ethics committee or the like, harmless 
from any claims for damages for unexpected injuries, including death, that may be caused by the 
participant’s participation in the trial but only to the extent that the claim is not caused by the fault or 
negligence of the participant(s) or Investigators. The Sponsor adheres to the guidelines of Medicines 
Australia for injury resulting from participation in a company sponsored trial, including the provision of 
‘No-fault clinical trial insurance’. 
 
 


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10.3 ABBREVIATIONS 
 
λ 
Elimination Rate Constant  
ACT 
 
Artemisinin Combination Therapy 
AE 
Adverse Event 
AESI 
Adverse Event of Special Interest 
ALP 
Alkaline Phosphatase 
ALT  
Alanine Aminotransferase 
AST 
Aspartate Aminotransferase 
ANZCTR 
Australia New Zealand Clinical Trial Registry 
anti-HBc Ab 
Anti-Hepatitis B Core Antibodies 
anti-HCV Ab 
Anti-Hepatitis C Virus Antibodies 
anti-HIV1 Ab 
Anti-Human Immunodeficiency Virus 1Antibodies 
anti-HIV2 Ab 
Anti-Human Immunodeficiency Virus 2 Antibodies 
AR 
Adverse Reaction 
AUC 
Area Under the Curve 
AUC0-∞ 
AUC to Infinite Time 
AUC0-last 
AUC to Last Quantifiable Concentration 
AUC0-t 
AUC to Last Measurable at t time 
Β-hCG 
Beta Human Chorionic Gonadotropin 
Blood Service  
Australian Red Cross Blood Service 
BMI 
Body Mass Index 
Cmax 
Maximum Plasma Concentration 
CL/F 
Apparent Oral Clearance 
CMI 
Consumer Medicine Information  
CRO 
Clinical Research Organisation 
CRU 
Clinical Research Unit 
CSR 
Clinical Study Report 
CTCAE 
Common Terminology Criteria for AEs 
DBP 
Diastolic Blood Pressure 
eCRF 
Electronic Case Report Form 
ECG 
Electrocardiographs 
EC50 
Half Maximal Effective Concentration 
EOS 
End of Study 
Emax 
Maximum effective concentration 
FBC 
Full Blood Count 
FDA 
Food and Drug Administration 
FSH 
Follicle-Stimulating Hormone 
G6PD 
Glucose-6-Phosphate Dehydrogenase  
GCP 
Good Clinical Practice 
HBs Ag 
Hepatitis B Surface Antigen 
HIV 
Human Immunodeficiency Virus 
HR 
Heart Rate 
HREC 
Human Research Ethics Committee  
IB 
Investigator’s Brochure 
IBSM 
Induced Blood Stage Malaria 


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79 
ICH 
International Conference on Harmonisation 
IMP 
Investigational Medicinal Product 
IV 
Intravenous  
LFT 
Liver Function Test 
MAP 
Modelling Analysis Plan 
MCB 
Master Cell Bank 
MedDRA 
Medical Dictionary for Regulatory Activities 
MIC 
Minimum Inhibitory Concentration 
MMV 
Medicines for Malaria Venture  
MPC90 
Minimal Parasiticidal Concentration 
NHMRC 
National Health and Medical Research Council 
NPA 
Nasopharyngeal Aspirate  
NTF 
Note To File 
PD 
Pharmacodynamics 
PICF 
Patient Information Consent Form 
PK 
Pharmacokinetics 
PK/PD 
Pharmacokinetics/Pharmacodynamics 
PRR 
Parasite Reduction Ratio 
PRR48 
Parasite Reduction Ratio in 48 hours 
PT 
Preferred Term 
Pt½ 
Parasite Clearance Half-Life 
QIMR Berghofer 
Queensland Institute of Medical Research Berghofer 
qPCR 
Quantitative Polymerase Chain Reaction 
qRT-PCR 
Quantitative Reverse-Transcriptase Polymerase Chain Reaction 
RAT 
Rapid Antigen Test 
RBCs 
Red Blood Cells 
RDW 
Red Blood Cell Distribution Width 
Rh 
Rhesus Antibody 
RSI 
Reference Study Information 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SARS-CoV-2 
Severe acute respiratory syndrome coronavirus 2  
SBP 
Systolic Blood Pressure 
SDRT 
Safety and Data Review Team 
SE 
Standard Error 
SOC 
System Organ Class 
SOP 
Standard Operating Procedure(s) 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
t½ 
Terminal Half-Life 
TGA 
Therapeutic Goods Administration 
Tmax 
Time taken to reach Cmax 
ULN 
Upper Limit of Normal 
Vd/F 
Apparent Volume Distribution 
WBC 
White Blood Cell 
WOCBP 
Women of Childbearing Potential  
WHO 
World Health Organisation 


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11 REFERENCES  
1. 
World Health Organization. World Malaria Report 2020. Geneva. 
2. 
Conrad MD, Rosenthal PJ. Antimalarial drug resistance in Africa: the calm before the storm? Lancet Infect 
Dis. 2019;19:e338-e51. 
3. 
Croft SL, Duparc S, Arbe-Barnes SJ, Craft JC, Shin CS, Fleckenstein L, et al. Review of pyronaridine anti-
malarial properties and product characteristics. Malar J. 2012;11:270. 
4. 
Bailly C. Pyronaridine: An update of its pharmacological activities and mechanisms of action. Biopolymers. 
2021;112:e23398. 
5. 
Burrows JN, Duparc S, Gutteridge WE, Hooft van Huijsduijnen R, Kaszubska W, Macintyre F, et al. New 
developments in anti-malarial target candidate and product profiles. Malar J. 2017;16:26. 
6. 
McCarthy JS, Baker M, O'Rourke P, Marquart L, Griffin P, Hooft van Huijsduijnen R, et al. Efficacy of OZ439 
(artefenomel) against early Plasmodium falciparum blood-stage malaria infection in healthy volunteers. J 
Antimicrob Chemother. 2016;71:2620-7. 
7. 
McCarthy JS, Lotharius J, Ruckle T, Chalon S, Phillips MA, Elliott S, et al. Safety, tolerability, 
pharmacokinetics, and activity of the novel long-acting antimalarial DSM265: a two-part first-in-human phase 
1a/1b randomised study. Lancet Infect Dis. 2017;17:626-35. 
8. 
McCarthy JS, Marquart L, Sekuloski S, Trenholme K, Elliott S, Griffin P, et al. Linking murine and human 
Plasmodium falciparum challenge models in a translational path for antimalarial drug development. Antimicrob 
Agents Chemother. 2016;60:3669-75. 
9. 
McCarthy JS, Ruckle T, Djeriou E, Cantalloube C, Ter-Minassian D, Baker M, et al. A Phase II pilot trial to 
evaluate safety and efficacy of ferroquine against early Plasmodium falciparum in an induced blood-stage malaria 
infection study. Malar J. 2016;15:469. 
10. 
Llanos-Cuentas A, Casapia M, Chuquiyauri R, Hinojosa JC, Kerr N, Rosario M, et al. Antimalarial activity of 
single-dose DSM265, a novel plasmodium dihydroorotate dehydrogenase inhibitor, in patients with uncomplicated 
Plasmodium falciparum or Plasmodium vivax malaria infection: a proof-of-concept, open-label, phase 2a study. 
Lancet Infect Dis. 2018;18:874-83. 
11. 
Phyo AP, Jittamala P, Nosten FH, Pukrittayakamee S, Imwong M, White NJ, et al. Antimalarial activity of 
artefenomel (OZ439), a novel synthetic antimalarial endoperoxide, in patients with Plasmodium falciparum and 
Plasmodium vivax malaria: an open-label phase 2 trial. Lancet Infect Dis. 2016;16:61-9. 
12. 
Investigator's Brochure: Pyronaridine Tetraphosphate Tablets. 2.0 ed: Shin Poong Pharm Co., Ltd. 
13. 
Pyramax 180 mg/60mg fim-coasted tablet: Summary of Product Characteristics. Seoul, South Korea: Shin 
Poong Pharmacweutical Co., Ltd. 
14. 
Ratcliff A, Siswantoro H, Kenangalem E, Maristela R, Wuwung RM, Laihad F, et al. Two fixed-dose 
artemisinin combinations for drug-resistant falciparum and vivax malaria in Papua, Indonesia: an open-label 
randomised comparison. Lancet. 2007;369:757-65. 
15. 
Fu S, Xiao SH. Pyronaridine: A new antimalarial drug. Parasitol Today. 1991;7:310-3. 
16. 
Chang C, Lin-Hua T, Jantanavivat C. Studies on a new antimalarial compound: pyronaridine. Trans R Soc 
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17. 
Shao BR. A review of antimalarial drug pyronaridine. Chin Med J (Engl). 1990;103:428-34. 
18. 
Ringwald P, Bickii J, Basco L. Randomised trial of pyronaridine versus chloroquine for acute uncomplicated 
falciparum malaria in Africa. Lancet. 1996;347:24-8. 
19. 
Looareesuwan S, Kyle DE, Viravan C, Vanijanonta S, Wilairatana P, Wernsdorfer WH. Clinical study of 
pyronaridine for the treatment of acute uncomplicated falciparum malaria in Thailand. Am J Trop Med Hyg. 
1996;54:205-9. 
20. 
Nieman AE, de Mast Q, Roestenberg M, Wiersma J, Pop G, Stalenhoef A, et al. Cardiac complication after 
experimental human malaria infection: a case report. Malar J. 2009;8:277. 
21. 
van Meer MP, Bastiaens GJ, Boulaksil M, de Mast Q, Gunasekera A, Hoffman SL, et al. Idiopathic acute 
myocarditis during treatment for controlled human malaria infection: a case report. Malar J. 2014;13:38. 
22. 
Watts RE, Odedra A, Marquart L, Webb L, Abd-Rahman AN, Cascales L, et al. Safety and parasite clearance 
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Australia. PLoS Med. 2020;17:e1003203. 
 
 
 


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12 APPENDIX 
Sponsor approved inclusion ranges  
Test 
Unit 
Low  
High 
Sodium 
mmol/L 
130 
150 
Potassium 
mmol/L 
3.0 
5.5 
Chloride 
mmol/L 
0.90 x LLN  
1.10 x ULN  
Calcium (Corr) 
mmol/L 
0.90 x LLN  
1.10 x ULN  
Urea 
mmol/L 
N/A 
1.75 x ULN 
Urate 
mmol/L 
N/A 
1.75 x ULN 
Creatinine 
umol/L 
N/A 
1.0 x ULN  
Creatine kinase 
U/L 
N/A 
< 2.5 x ULN 
eGFR 
mL/min/1.7 3m2 60  
N/A 
Glucose Fasted 
mmol/L 
N/A 
1.0 x ULN 
Total Bilirubin 
umol/L 
N/A 
1.0 x ULN 
ALP 
U/L 
N/A  
1.5 x ULN 
AST 
U/L 
N/A 
1.0 x ULN 
ALT 
U/L 
N/A 
1.0 x ULN 
GGT 
U/L 
N/A 
1.0 x ULN 
Cholesterol 
mmol/L 
N/A 
1.2 x ULN  
HDL Cholesterol 
mmol/L 
0.9x LLN  
N/A 
LDL Cholesterol 
mmol/L 
N/A 
1.25 x ULN  
Hb 
g/L 
0.9x LLN 
 
1.1.x ULN  
Plats  
x109/L 
0.9x LLN  
1.1 x ULN  
WCC 
x109/L 
0.9x LLN  
1.1 x ULN  
Neuts 
x109/L 
1.0 x LLN  
1.1 x ULN  
Lymphs 
x109/L 
1.0 x LLN  
1.1 x ULN  
Protein (dipstick) 
  
N/A 
1+ or 30mg/dL 
Ketones (dipstick) 
  
N/A 
<3+ or <80mg/dL 
Red Blood Cells (MCS) 
  
N/A 
<20* 
*A result ≥ 20 is acceptable 
for female participants 
currently menstruating. 
White Blood Cells 
(MCS) 
  
N/A 
<10 
Casts (MCS) 
  
N/A 
<2/high power field 


Pyronaridine VIS Protocol Version 4.0; 27 July 2022 
 
 
83 
 

SAP_001.pdf: 
Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 1 of 4 9  
S T A TI S TI C A L A N A L Y SI S P L A N  
A p h a s e 1 b st u d y t o c h ar a ct eri s e t h e p h ar m a c o ki n eti c/ p h ar m a c o d y n a mi c 
r el ati o n s hi p of p yr o n ari di n e i n h e alt h y a d ult V ol u nt e er s e x p eri m e nt all y i nf e ct e d 
wit h bl o o d st a g e Pl a s m o di u m f al ci p ar u m . 
Pr ot o c ol N o.: M M V _ P yr o n ari di n e _ 2 1 _ 0 1  
Pr o d u ct C o d e: P yr o n ari di n e  
P R E P A R E D F O R:    M e di ci n e s f or M al ari a V e nt ur e ( M M V)  
   I C C- Bl o c k G 3r d Fl o or 
   2 0, R o ut e d e Pr é -B oi s  
   1 2 1 5 G e n e v a 1 5  
   S wit z erl a n d 
   P h o n e: + 4 1 2 2 5 5 5 0 3 0 0  
P R E P A R E D B Y:  
 S o ut h er n St ar R e s e ar c h 
L e v el 1, 1 M erri w a St 
G or d o n, N S W, 2 0 7 2   
A u str ali a   
D A T E O F I S S U E:     0 1 D E C 2 0 2 2  (V 1. 0 ); 0 6 F E B 2 0 2 3 ( V 1. 1) 
V E R SI O N/ S T A T U S:  V er si o n 1. 1  
      A U T H O R: 
L o c a di a h K u w a n d a  


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M yri a m El G a al o ul
R e a s o n: I a p pr o v e t hi s
d o c u m e nt.
D at e: F e b 1 6, 2 0 2 3 0 8: 5 9
G M T + 1
1 6- F e b- 2 0 2 3
El e ctr o ni c all y si g n e d b y:
L o c a di a h K u w a n d a
R e a s o n: I h a v e a ut h or e d
t hi s d o c u m e nt.
D at e: F e b 1 7, 2 0 2 3
1 5: 1 4 G M T + 1 1
1 7- F e b- 2 0 2 3


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 of 4 9  
 
T a bl e of c o nt e nts  
 
1.  
A B B R E VI A TI O NS  .............................................................................................................. 5 
2.  
I N T R O D U C TI O N ............................................................................................................... 7 
3.  
P R OJ E C T O V E R VI E W  ........................................................................................................ 8 
3. 1  
St u dy D esi g n  ........................................................................................................................ 8 
3. 2  
Sc h e m a  .............................................................................................................................. 1 0  
3. 3  
Sc h e d ul e of Acti viti es  ......................................................................................................... 1 1  
3. 4  
O bj ecti v es  .......................................................................................................................... 1 4  
3. 5  
E n d p oi nts  ........................................................................................................................... 1 4  
3. 6  
S a m pl e Siz e  ........................................................................................................................ 1 5  
3. 7  
Tr e at m e n t Assi g n m e nt a n d R a n d o mis ati o n ........................................................................ 1 5  
4.  
S T A TI S TI C AL C O NSI D E R A TI O NS ...................................................................................... 1 6  
4. 1  
D at a C a pt ur e  ...................................................................................................................... 1 6  
4. 2  
St atisti c al Pr o gr a m mi n g ..................................................................................................... 1 7  
5.  
A N AL YSI S S E TS ............................................................................................................... 1 9  
5. 1   
A n al ysis S et D escri pti o ns .................................................................................................... 1 9  
6.  
P R O T O C OL D E VI A TI O NS ................................................................................................ 1 9  
7.  
P artici p a nt DIS P OSI TI O N  ............................................................................................... 2 0  
Dis p ositi o n  .................................................................................................................................... 2 0  
8.  
D E M O G R A P HI C A N D B A S ELI N E I N F O R M A TI O N .............................................................. 2 0  
8. 1   
 D e m o gr a p hics  ................................................................................................................... 2 0  
8. 2  
M e dic al Hist or y  .................................................................................................................. 2 0  
8. 3  
Pri o r M e dic ati o ns ............................................................................................................... 2 0  
8. 4  
Dr u g Scr e e n  ....................................................................................................................... 2 1  
8. 5  
Alc o h ol Scr e e n  ................................................................................................................... 2 1  
8. 6  
S A R S -C o V -2 Scr e e ni n g  ........................................................................................................ 2 1  
8. 7  
F ollicl e Sti m ul ati ng H or m o n e ( F S H)  .................................................................................... 2 1  
8. 8  
B eck  D e pr essi o n I nv e nt or y  ................................................................................................. 2 1  
9.  
T R E A T M E N T E X P O S U R E  ................................................................................................. 2 2  
1 0.  
P RI M A R Y E N D P OI N T A N AL YSI S ( P K/ P D A n al ysis) ........................................................ 2 2  
1 0. 1  
Ov er vi e w of m et h o d ol o g y f or P K/ P D M o d elli n g ................................................................. 2 2  
1 0. 2  
S o urc es of D at a  .................................................................................................................. 2 2  
1 0. 3  
P h a r m ac oki ni etics .............................................................................................................. 2 2  
1 0. 4  
P h ar m ac o dy n a mics  ............................................................................................................ 2 2  


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 4 of 4 9  
 
1 0. 5  
D at a Ass e m bl y  ................................................................................................................... 2 3  
1 0. 6.  
M o d el b uil di n g  ............................................................................................................... 2 4  
1 0. 7.   
 D eri v ati o n of s ec o n d ar y effic acy p ar a m et ers ................................................................. 2 7  
1 0. 8.   
 P ost -h oc A n al ysis ........................................................................................................... 2 7  
Si m ul ati o ns of cli nic al effic acy p ar a m et ers a n d e n d p oi nts i n Afric a n c hil dr e n  ................................ 2 7  
1 0. 9.   
 Pr es e nt ati o n of R es ults  .................................................................................................. 2 8  
1 1.  
S E C O N D A R Y E N D P OI N TS A N AL YSI S ............................................................................ 2 8  
1 1. 1  
P D A n al ysis ......................................................................................................................... 2 8  
1 1. 1. 2   
 P ar asit e cl e ar a nc e ki n etics  ............................................................................................. 2 9  
1 1. 2  ............................................................................................................................................... 3 2  
P K A n al ysis  .................................................................................................................................... 3 2  
1 1. 3  ............................................................................................................................................... 3 5  
S af ety  ............................................................................................................................................ 3 5  
1 1. 3. 1  ............................................................................................................................................ 3 5  
A dv ers e Ev e nts .............................................................................................................................. 3 5  
1 1. 3. 2  
C o nc o mit a nt M e dic ati o n  ................................................................................................ 3 7  
1 1. 3. 3  
L a b or at or y  ..................................................................................................................... 3 8  
1 1. 4  
Vit al Si g ns  .......................................................................................................................... 4 1  
1 1. 5  
B o dy M e as ur e m e nts  .......................................................................................................... 4 1  
P hysic al Ex a mi n ati o n  ..................................................................................................................... 4 2  
1 1. 6  
1 2 -l e a d E C G ....................................................................................................................... 4 2  
1 1. 7  
Pr es e nt ati o n of S af ety R es ults  ............................................................................................ 4 3  
1 2.  
Ex pl or at ory P ar a m et ers  ............................................................................................. 4 3  
1 3.  
C H A N G ES T O T H E PL A N N E D A N AL YSI S ....................................................................... 4 4  
1 4.  
I N T E RI M A N D FI N AL A N AL YSI S ................................................................................... 4 4  
1 5.  
S O F T W A R E  ................................................................................................................ 4 4  
1 6.  
R E F E R E N C ES  .............................................................................................................. 4 4  
1 7.  
A P P E N DI X  .................................................................................................................. 4 6  
T a bl e A. P K/ P D P l a n n e d Fi g ur es a n d T a bl es ........................................ Err or! B o o k m ar k n ot d efi n e d.  
T a bl e B. P D Pl a n n e d T a bl es, Fi g ur es a n d Listi n gs  ............................................................................ 4 6  
T a bl e C. P h ar m ac oki n etic P ar a m et ers a n d R e p orti n g S p ecific ati o ns  .... Err or! B o o k m ar k n ot d efi n e d.  
T a bl e D. B as eli n e C h ar act eristics a n d S af ety A n al ysis T a bl es a n d Listi n gs  ....................................... 4 6  
 
 
 
 
D el
D el


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 5 of 4 9  
 
1.  A B B R E VI A TI O N S  
A D a M  
A n al y si s D at a M o d el  
A E  
A d v er s e E v e nt  
A E SI  
A d v er s e E v e nt of S p e ci al I nt er e st  
A L P  
Al k ali n e p h o s p h at a s e  
A L T  
Al a ni n e A mi n otr a n sf er a s e  
A P R 2 8  
A d e q u at e P ar a sit ol o gi c al R e s p o n s e R at e at D a y 2 8  
A P T T  
A cti v at e d p arti al t hr o m b o pl a sti n ti m e  
A S T  
A s p art at e A mi n otr a n sf er a s e  
A T C  
A n at o mi c al T h er a p e uti c C h e mi c al  
A U C  
Ar e a U n d er t h e Bl o o d  C o n c e ntr ati o n Ti m e C ur v e 
A U C l a st 
Ar e a u n d er t h e c ur v e fr o m ti m e 0 t o l a st m e a s ur a bl e c o n c e ntr ati o n  
A U C i nf 
Ar e a u n d er t h e c ur v e fr o m ti m e 0 t o i nfi nit y  
β- H C G  
β- H u m a n C h ori o ni c G o n a d otr o pi n  
B DI  
B e c k D e pr e s si o n I n v e nt or y  
BI C  
B a y e si a n I nf or m ati o n Crit eri a 
B L Q  
B el o w t h e Li mit of Q u a ntifi c ati o n 
B MI  
B o d y M a s s I n d e x  
C DI S C  
Cli ni c al D at a I nt er c h a n g e St a n d ar d s C o n s orti u m  
CI  
C o nfi d e n c e I nt er v al  
C K  
Cr e ati n e Ki n a s e  
C L/ F  
A p p ar e nt Or al Cl e ar a n c e  
C m a x  
M a xi m u m bl o o d  c o n c e ntr ati o n o b s er v e d 
C O VI D -1 9  
C or o n a vir u s di s e a s e 2 0 1 9  
C R A  
Cli ni c al R e s e ar c h A s s o ci at e  
C R F  
C a s e R e p ort F or m  
C S R  
Cli ni c al St u d y R e p ort  
C T C A E  
C o m m o n T er mi n ol o g y Crit eri a f or A d v er s e E v e nt s  
D B P  
Di a st oli c Bl o o d Pr e s s ur e  
D T S  
D at a Tr a n sf er S p e cifi c ati o n  
E C 5 0  
C o n c e ntr ati o n at 5 0 % of M a xi m u m Eff e ct 
E C G  
El e ctr o c ar di o gr a p h y  
e C R F  
El e ctr o ni c C a s e R e p ort F or m  
E D C  
El e ctr o ni c D at a c a pt ur e  
E M A X  
M a xi m u m Eff e ct 
E O S  
E n d of St u d y  
F A S  
F ull A n al y si s S et  
F S H  
F olli c ul ar Sti m ul ati n g H or m o n e  
G 6 P D  
Gl u c o s e- 6- p h o s p h at a s e  
G G T  
G a m m a Gl ut a m yl Tr a n s p e pti d a s e  
H B s A g  
H e p atiti s B S urf a c e A nti g e n  
H E E N T  
H e a d, E y e s, E ar s, N o s e, T hr o at  
HI V  
H u m a n I m m u n o d efi ci e n c y Vir u s  
H R  
H e art R at e  
I B S M 
I n d u c e d Bl o o d St a g e M al ari a 
I C 5 0 
I n v e sti g ati o n al M e di ci n al Pr o d u ct 
II V 
I nt er I n di vi d u al V ari a bilit y 
I M P 
I n v e sti g ati o n al M e di ci n al Pr o d u ct 
I N R 
I nt er n ati o n al N or m ali z e d R ati o 
L D H  
L a ct at e D e h y dr o g e n a s e  
L L O Q  
L o w er Li mit of Q u a ntifi c ati o n 
L O Q  
Li mit of Q u a ntifi c ati o n  
M e d D R A  
M e di c al Di cti o n ar y f or R e g ul at or y A cti viti e s  
MI C  
Mi ni m u m I n hi bit or y C o n c e ntr ati o n  
M M V  
M e di ci n e f or M al ari a V e nt ur e  
M P C 9 0  
Mi ni m al P ar a siti ci d al C o n c e ntr ati o n  
N C A  
N o n - C o m p art m e nt al A n al y si s 
N D  
N o n D et e ct 
N L M E  
N o nli n e ar Mi x e d Eff e ct s 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 6 of 4 9  
 
P C  
P h ar m a c o ki n eti c s C o n c e ntr ati o n  
P C S A  
P ot e nti al Cli ni c all y Si g nifi c a nt A b n or m alit y  
P D  
P h ar m a c o d y n a mi c s  
Pf i R B C  
P.f al ci p ar u m I nf e ct e d R e d Bl o o d C ell 
P K  
P h ar m a c o ki n eti c s  
P K P D  
P h ar m a c o ki n eti c- p h ar m a c o d y n a mi c  
P N  
Pr ef err e d N a m e  
P R R 4 8  
P ar a sit e R e d u cti o n R ati o O v er 4 8 h o ur s  
P T  
Pr ef err e d T er m  
P Ct 1/ 2 
P ar a sit e Cl e ar a n c e H alf- Lif e  
QI M R B  
QI M R B er g h of er M e di c al R e s e ar c h I n stit ut e  
q P C R  
Q u a ntit ati v e P ol y m er a s e C h ai n R e a cti o n  
q R T -P C R  
Q u a ntit ati v e R e v er s e Tr a n s cri pt a s e P ol y m er a s e C h ai n R e a cti o n 
Q T c B  
C orr e ct e d Q T i nt er v al wit h B a z ett’ s F or m ul a  
Q T c F  
C orr e ct e d Q T i nt er v al wit h Fri d eri ci a’ s F or m ul a  
R A T  
R a pi d A nti g e n T e st  
S A E  
S eri o u s A d v er s e E v e nt  
S A E M  
St o c h a sti c A p pr o xi m ati o n E x p e ct ati o n M a xi mi z ati o n 
S A P  
St ati sti c al A n al y si s Pl a n  
S A S  
St ati sti c al A n al y si s S y st e m  
S B P  
S y st oli c Bl o o d Pr e s s ur e  
S B Q  
S wi s s Bi o Q u a nt C e ntr al  
S D  
St a n d ar d D e vi ati o n  
S D T M  
St u d y D at a T a b ul ati o n M o d el  
S E  
St a n d ar d Err or  
S O C  
S y st e m Or g a n Cl a s s  
S O P  
St a n d ar d O p er ati n g Pr o c e d ur e  
S R C  
S af et y R e vi e w C o m mitt e e  
S S R  
S o ut h er n St ar R e s e ar c h  
t 1/ 2 
H alf -Lif e  
T E A E  
Tr e at m e nt E m er g e nt A d v er s e E v e nt  
T m a x  
Ti m e t o R e a c h M a xi m u m Bl o o d  C o n c e ntr ati o n 
VI S  
V ol u nt e er I nf e cti o n St u d y  
V s s/ F  
A p p ar e nt V ol u m e of Di stri b uti o n at t h e St e a d y St at e Aft er Si n gl e D o s e  
V P C  
Vi s u al Pr e di cti v e C h e c k 
V z/ F  
A p p ar e nt V ol u m e of Di stri b uti o n aft er Si n gl e D o s e  
W O C B P  
W o m e n of C hil d B e ari n g P ot e nti al    
W H O Dr u g  
W orl d H e alt h Or g a ni z ati o n Dr u g Di cti o n ar y  
 
 
 
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 7 of 4 9  
 
2.  I N T R O D U C TI O N 
T h e f oll o wi n g St ati sti c al A n al y si s Pl a n ( S A P) pr o vi d e s t h e o utli n e f or t h e st ati sti c al a n al y si s of 
t h e d at a fr o m t h e M M V _ P yr o n ari di n e _ 2 1 _ 0 1  st u d y . T h e S A P d et ail s  t h e m et h o d s f or st ati sti c al 
a n al y si s of o ut c o m e s r el ati n g t o s af et y, p h ar m a c o ki n eti c s ( P K), p h ar m a c o d y n a mi c s ( P D), a n d 
p o p ul ati o n  m o d elli n g of p h ar m a c o ki n eti c  / p h ar m a c o d y n a mi c  ( P K/ P D) d at a ari si n g  fr o m  t hi s 
cli ni c al  tri al.  T hi s  S A P  h a s  b e e n  d e v el o p e d  a n d  fi n ali s e d  pri or  t o  d at a b a s e  l o c k  a n d  fi n al 
a n al y si s.  
 
Pl a n n e d  a n al y s e s  i d e ntifi e d  i n  t hi s  S A P  will  b e  i n cl u d e d  i n t h e cli ni c al  st u d y  r e p ort  ( C S R), 
r e g ul at or y s u b mi s si o n s, a n d f ut ur e m a n u s cri pt s. I n a d diti o n, p o st h o c e x pl or at or y a n al y s e s n ot 
n e c e s s aril y  i d e ntifi e d  i n  t hi s  S A P  m a y  b e  p erf or m e d  t o  f urt h er  e x a mi n e  st u d y  d at a.    A n y 
p o st h o c, or u n pl a n n e d, e x pl or at or y a n al y s e s p erf or m e d will b e cl e arl y i d e ntifi e d a s s u c h i n t h e 
fi n al C S R. 
 
A n y si g nifi c a nt c h a n g e s fr o m pl a n n e d a n al y s e s will al s o b e d e s cri b e d i n t h e fi n al C S R.    
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 8 of 4 9  
 
3.  P R O J E C T O V E R VI E W  
3. 1  St u d y D e si g n  
T hi s  w a s  a  si n gl e- c e ntr e,  o p e n-l a b el  p h a s e 1 b  st u d y u si n g  t h e P.  f al ci p ar u m  i n d u c e d bl o o d 
st a g e m al ari a (I B S M) m o d el t o c h ar a ct eri s e t h e P K/ P D pr ofil e of p yr o n ari di n e.  
 
U p t o 1 8 h e alt h y, m al ari a n aï v e a d ult p arti ci p a nt s ar e pl a n n e d t o b e e nr oll e d i nt o t hi s st u d y, i n 
c o h ort s of u p t o si x p arti ci p a nt s e a c h. F oll o wi n g a s cr e e ni n g p eri o d of u p t o 2 8 d a y s, c o h ort s 
of u p t o 6 h e alt h y p arti ci p a nt s will b e e nr oll e d. Aft er c o nfir m ati o n of eli gi bilit y, i n cl u di n g a 
R a pi d A nti g e n T e st ( R A T) f or S A R S- C o V - 2, e a c h p arti ci p a nt will b e i n o c ul at e d i ntr a v e n o u sl y 
o n D a y 0 wit h a p pr o xi m at el y 2, 8 0 0 vi a bl e P. f al ci p ar u m -i nf e ct e d er yt hr o c yt e s. P arti ci p a nt s will 
b e f oll o w e d u p d ail y vi a p h o n e c all or t e xt m e s s a g e o n D a y s 1 t o 3 p o st-i n o c ul ati o n t o s oli cit 
a n y a d v er s e e v e nt s. 
 
P arti ci p a nt s will att e n d t h e cli ni c al u nit o n c e o n D a y s 4, 5, 6 a n d 7 f or cli ni c al e v al u ati o n a n d 
bl o o d s a m pli n g t o m o nit or t h e pr o gr e s si o n of p ar a sit a e mi a, u si n g q u a ntit ati v e p ol y m er a s e 
c h ai n r e a cti o n ( q P C R) t ar g eti n g t h e g e n e e n c o di n g 1 8 S r R N A (r ef err e d t o h er e aft er a s 
m al ari a 1 8 S q P C R).  
 
P arti ci p a nt s will h a v e a n a s o p h ar y n g e al a s pir at e ( N P A) f or S A R- C o V -2 o n D a y 6 a m , a n d 
bl o o d c oll e ct e d o n D a y 7 a m t o m o nit or h a e m at ol o g y a n d bi o c h e mi str y.  
 
P arti ci p a nt s will b e a d mitt e d t o t h e cli ni c al tri al u nit o n D a y 8 f or d o si n g wit h t h e 
i n v e sti g ati o n al m e di ci n al pr o d u ct (I M P; p yr o n ari di n e) w h e n p ar a sit a e mi a f or t h e m aj orit y of 
p arti ci p a nt s i s e x p e ct e d t o b e a b o v e 5, 0 0 0 p ar a sit e s/ m L. P arti ci p a nt s will n e e d t o h a v e a 
n e g ati v e S A R S -C o V -2 R A T pri or t o a d mi s si o n. 
 
P yr o n ari di n e will b e a d mi ni st er e d a s a si n gl e or al d o s e. Diff er e nt d o s e s of p yr o n ari di n e will b e 
a d mi ni st er e d a cr o s s a n d wit hi n c o h ort s i n or d er t o eff e cti v el y c h ar a ct eri s e t h e P K/ P D 
r el ati o n s hi p. E a c h c o h ort will c o m pri s e u p t o t hr e e d o s e gr o u p s, wit h p arti ci p a nt s r a n d o mi s e d 
t o a d o s e gr o u p o n t h e d a y of d o si n g. T h e hi g h e st d o s e of p yr o n ari di n e a d mi ni st er e d will b e 
n o m or e t h a n 7 2 0 m g; t h e l o w e st d o s e a d mi ni st er e d will b e n o l e s s t h a n 1 8 0 m g.  
 
T h e pr o p o s e d d o si n g r e gi m e n f or t h e C o h ort 1 a s s u mi n g si x p arti ci p a nt s ar e e nr oll e d, i s a s 
b el o w. If l e s s t h a n si x P arti ci p a nt s ar e e nr oll e d t h e S af et y D at a R e vi e w T e a m ( S D R T) will 
m e et b et w e e n D a y 0 a n d D a y 8, t o d e ci d e o n t h e d o s e/ s t o b e a d mi ni st er e d. 
 
 
D o s e gr o u p  
 
1 A ( n = 2)  
1 B ( n = 2)  
1 C ( n = 2)  
P yr o n ari di n e  
3 6 0 m g  
5 4 0 m g  
7 2 0 m g  
 
 
 
 
 
T h e  S D R T  will  r e vi e w s af et y, t ol er a bilit y, p ar a sit a e mi a a n d P K/ P D d at a t o d et er mi n e t h e d o s e 
t o b e a d mi ni st er e d t o C o h ort 2.  
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 9 of 4 9  
 
T h e  st u d y  will  c o n cl u d e  w h e n  s uffi ci e nt  d at a  h a v e  b e e n  o bt ai n e d  t o  d efi n e  t h e  P K/ P D 
p ar a m et er s f or p yr o n ari di n e (t h e pri m ar y e n d p oi nt).    
A  m a xi m u m  s a m pl e  si z e  of  1 8  P arti ci p a nt s  a d mi ni st er e d  p yr o n ari di n e  i s  e x p e ct e d  t o  b e 
s uffi ci e nt t o a c hi e v e t h e pri m ar y e n d p oi nt. 
 
P arti ci p a nt s will b e c o nfi n e d i n t h e cli ni c al u nit f or at l e a st 9 6 h ( D a y s 8 - 1 2) t o m o nit or t h e 
s af et y a n d t ol er a bilit y of p yr o n ari di n e d o si n g a n d t o e n s ur e a d e q u at e cli ni c al r e s p o n s e 
a g ai n st P. f al ci p ar u m . D uri n g c o nfi n e m e nt, r e g ul ar s af et y a s s e s s m e nt s will b e p erf or m e d, a n d 
bl o o d will b e c oll e ct e d t o m o nit or p ar a sit e cl e ar a n c e a n d p yr o n ari di n e c o n c e ntr ati o n.  
 
All p arti ci p a nt s will r e c ei v e c o m p ul s or y a nti m al ari al r e s c u e tr e at m e nt wit h Ri a m et ® o n D a y 
4 7 ± 2 or e arli er i n t h e f oll o wi n g c a s e s: 
 
•   
P ar a sit a e mi a i s n ot r e d u c e d b y a p pr o xi m at el y 1 0-f ol d at t h e e n d of t h e 9 6-h o ur 
c o nfi n e m e nt p eri o d ( D a y 1 2) w h e n c o m p ar e d wit h p e a k p ar a sit a e mi a. 
•   
P ar a sit e r e gr o wt h o c c ur s aft er a n i niti al r e d u cti o n i n p ar a sit a e mi a, wit h r e gr o wt h 
d efi n e d a s ≥5 0 0 0 p ar a sit e s/ m L a n d a t w o -f ol d p ar a sit a e mi a i n cr e a s e wit hi n 4 8 h o ur s.  
•   
P arti ci p a nt di s c o nti n u ati o n/ wit h dr a w al fr o m t h e st u d y. 
•  
Ri a m et ®  tr e at m e nt m a y al s o b e  i niti at e d at a n y ti m e at t h e I n v e sti g at or’ s di s cr eti o n i n 
t h e i nt er e st of p arti ci p a nt s af et y. 
 
P ar a sit e lif e c y cl e st a g e s (i n cl u di n g g a m et o c yt e s) will b e m o nit or e d at s el e ct ti m e p oi nt s 
d uri n g t h e st u d y b y c oll e cti n g bl o o d s a m pl e s a n d u si n g q u a ntit ati v e r e v er s e-tr a n s cri pt a s e 
p ol y m er a s e c h ai n r e a cti o n ( q R T- P C R) t ar g eti n g p ar a sit e lif e c y cl e st a g e- s p e cifi c m R N A 
tr a n s cri pt s. 
 
F or  C o h ort  1,  if  g a m et o c yt e s ar e   pr e s e nt at  or  aft er  t h e  ti m e  of  tr e at m e nt  wit h  Ri a m et ®, 
Pri m a ci n ®  ( pri m a q ui n e) w ill  b e  a d mi ni st er e d  a s  a  si n gl e  or al  d o s e  at  t h e  I n v e sti g at or’ s 
di s cr eti o n.  Fr o m C o h ort 2 o n w ar d s , a si n gl e d o s e of Pri m a ci n ® will b e a d mi ni st er e d at a n y 
ti m e p oi nt aft er t h e c o nfi n e m e nt p eri o d w h e n g a m et o c yt e s ar e  d et e ct e d a s d et er mi n e d b y q R T -
P C R, at t h e I n v e sti g at or’ s di s cr eti o n.  
 
If a n all er g y or c o ntr ai n di c ati o n t o Ri a m et ® d e v el o p s, M al ar o n e ® ( at o v a q u o n e/ pr o g u a nil) will 
b e a d mi ni s t er e d. If i n t h e r ar e e v e nt p arti ci p a nt s ar e u n a bl e t o c o m pl et e or al a nti m al ari al r e s c u e 
tr e at m e nt, t h e y will b e a d mitt e d t o h o s pit al t o r e c ei v e i ntr a v e n o u s Art e s u n at e . 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 0  of 4 9  
 
 
 
 
3. 2  
S c h e m a  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
M al ari a c h all e ng e 
a g e nt a d mi nistr ati o n  
D a y -2 8                  D a y 0                                   D a y 8         D a y 1 2                            D a y 5 0 ± 2  
P yr o n ari di n e 
d osi n g  
Scr e e ni n g 
O ut p ati e nt  
m al ari a  
Cli nic 
c o nfi n e m e nt 
O ut p ati e nt 
f oll o w-u p  
E n d of 
st u d y 
     A nti m al ari al r esc u e 
tr e at m e nt ( D ay 4 7± 2 or 
e arli er if r e q uir e d)  
St u d y 
Ti m eli n e  


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 1  of 4 9  
 
3. 3  S c h e d ul e of A cti viti e s  
T h e b el o w t a bl e s u m m ari s e s t h e a cti viti e s a n d pr o c e d ur e s t o b e c o n d u ct e d a s p er t hi s pr ot o c ol d uri n g s cr e e ni n g, eli gi bilit y, m al ari a i n o c ul ati o n, o ut 
p ati e nt m o nit ori n g, c o nfi n e m e nt, a n d at t h e e n d of st u d y. 
 
 
Scr e e ni n g 
Eli gi bility a 
M al ari a 
i n ocul ati o n  
P h o n e 
c o nt act 
M al ari a 
m o nit ori ng
b 
I M P a d mi nistr ati o n a n d cli nic al 
u nit c o nfi n e m e nt c 
O ut -p ati e nt 
m o nit ori ng
d 
E O S  
D ay  
-2 8 t o - 1 
-3 t o - 1 
0 
1 t o 3  
4 t o 7  
8 
9 
1 0  
1 1  
1 2  
1 3 t o 5 0 ± 2  
5 0 ± 2  
Eli gi bility ass ess m e nts 
I nf or m e d c o ns e nt 
X 
 
 
 
 
 
 
 
 
 
 
 
B ec k D e pr essi o n I nv e nt ory- II 
X 
 
 
 
 
 
 
 
 
 
 
 
C ar di ov asc ul ar risk  
X 
 
 
 
 
 
 
 
 
 
 
 
D e m ogr a p h y  
X 
 
 
 
 
 
 
 
 
 
 
 
M e dic al a n d s oci al hist or y, 
i nc/ exc. crit eri a, & pri or 
m e dic ati o ns  
X 
 
X 
 
 
 
 
 
 
 
 
 
Dr u g & alc o h ol scr e e n  
X 
 
X 
 
 
X 
 
 
 
 
 
 
B o d y h ei g ht  
X 
 
 
 
 
 
 
 
 
 
 
 
B o d y w ei g ht  
X 
 
 
 
 
 
 
 
 
 
 
X 
S er ol o gy & R B C all o a nti b o dy  
X 
 
 
 
 
 
 
 
 
 
 
X 
C o a g ul ati o n pr ofil e  
X 
 
 
 
 
 
 
 
 
 
 
 
C O VI D -1 9 t esti n g l 
 
X 
X 
 
X ( D 6 a m) 
X 
 
 
 
 
 
 
G 6 P D  
X 
 
 
 
 
 
 
 
 
 
 
 
S er u m pr e g n a nc y t est ( all 
f e m al es at scr e e ni n g & W O C B P 
at E O S)  
X 
 
 
 
 
 
 
 
 
 
 
X 
Uri n e pr e g n a nc y t est ( W O C B P) 
 
 
X 
 
 
X 
 
 
 
 
 
 
F S H ( p ost -m e n o p a us al w o m e n)  X 
 
 
 
 
 
 
 
 
 
 
 
S af ety ass ess m e nts  
F ull p h ysic al ex a m  
X 
 
 
 
 
 
 
 
 
 
 
X 
A b br e vi at e d p hysic al ex a m  
 
 
X 
 
 
X 
 
 
 
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 2  of 4 9  
 
 
Scr e e ni n g 
Eli gi bility a 
M al ari a 
i n ocul ati o n  
P h o n e 
c o nt act 
M al ari a 
m o nit ori ng
b 
I M P a d mi nistr ati o n a n d cli nic al 
u nit c o nfi n e m e nt c 
O ut -p ati e nt 
m o nit ori ng
d 
E O S  
D ay  
-2 8 t o - 1 
-3 t o - 1 
0 
1 t o 3  
4 t o 7  
8 
9 
1 0  
1 1  
1 2  
1 3 t o 5 0 ± 2  
5 0 ± 2  
Sy m pt o m -dir ect e d p h ysic al 
ex a m e 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
 
Vit al si g ns f 
X 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
E C G  
X 
 
X 
 
 
X 
X 
X 
X 
X 
X 
X 
Uri n al ysis  
X 
X 
 
 
 
 
 
 
 
 
 
X 
H a e m. & Bi oc h e m.  
X  g 
X 
 
 
X ( D 7 a m) 
X 
 
 
X 
 
X 
X 
S af et y s er u m r et e nti o n  
 
 
X 
 
 
 
 
 
 
 
 
X 
Di ar y c ar d  
 
 
X 
X 
X 
 
 
 
 
 
X 
X 
A Es & C o n m e ds.  
 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
St u d y i nt er v e nti o ns 
C h all e ng e ag e nt i n oc ul ati o n  
 
 
X 
 
 
 
 
 
 
 
 
 
P yr o n ari di n e a d mi nistr ati o n  
 
 
 
 
 
X 
 
 
 
 
 
 
D efi nitiv e/r esc u e a nti m al ari al  
tr e at m e nt 
 
 
 
 
 
 
 
 
 
 
X h 
 
M al ari a m o nit ori n g a n d P K bl o o d s a m pli n g  
M al ari a 1 8 S q P C R
 i 
 
 
X 
 
X 
X 
X 
X 
X 
X 
X 
X 
P ar asit e lif ec ycl e st a g e q R T -
P C R bl o o d s a m pli n g  
 
 
 
 
 
 
 
 
 
 
X 
 
P yr o n ari di n e c o nc e ntr ati o n j  
 
 
 
 
 
X 
X 
X 
X 
X 
X 
X 
Ot h er  
M al ari a f ut ur e r es e arc h 
( o pti o n al) 
 
 
X 
 
 
X 
 
 
 
 
X m  
X 
P ar asit e vi a bility ex viv o  
gr o wt h k  
 
 
 
 
 
X 
X 
X 
X 
X 
X ( p ar asit e 
r e gr o wt h o nl y) 
 
A E s:  a d v er s e  e v e nt s;  E C G:  el e ctr o c ar di o gr a p h;  E O S:  E n d  of  St u d y;  G 6 P D:  gl u c o s e- 6- p h o s p h at e  d e h y dr o g e n a s e  q P C R:  q u a ntit ati v e  p ol y m er a s e  c h ai n  r e a cti o n;  q R T- P C R: 
q u a ntit ati v e r e v er s e-tr a n s cri pt a s e p ol y m er a s e c h ai n r e a cti o n; R B C: r e d bl o o d c ell; W O C B P: w o m e n of c hil d b e ari n g p ot e nti al. 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 3  of 4 9  
 
a  T hi s vi sit i s n ot r e q uir e d i n t h e e v e nt t h at t h e s cr e e ni n g vi sit i s c o n d u ct e d wit hi n t hi s p eri o d. 
b  D ail y vi sit s. 
c S e e S e cti o n 8. 1. 5 f or d et ail s of a s s e s s m e nt s t o b e p erf or m e d pri or t o a n d aft er p yr o n ari di n e a d mi ni str ati o n. Al s o n ot e t h at t h er e ar e m ulti pl e s a m pl e s/ a s s e s s m e nt s f or s o m e a cti viti e s 
o n t h e c o nfi n e m e nt d a y s.  
d F oll o w u p vi sit s will o c c ur at l e a st t hr e e ti m e s p er w e e k. S e e S e cti o n 8. 1. 6 f or t h e s p e cifi c ti mi n g s of diff er e nt a cti viti e s.  
e S y m pt o m- dir e ct e d p h y si c al e x a m will o nl y b e p erf or m e d if cli ni c all y i n di c at e d. 
f S u pi n e & st a n di n g bl o o d pr e s s ur e a n d h e art r at e r e q uir e d at s cr e e ni n g; s e at e d m e a s ur e m e nt s o nl y at ot h er ti m e - p oi nt s. O n D a y 0, r e c or d vit al si g n s pri or t o i n o c ul ati o n a n d pri or t o 
l e a vi n g t h e cli ni c al tri al u nit ( a p pr o xi m at el y 6 0 mi n ut e s aft er i n o c ul ati o n). R e c or d vit al si g n s 3 ti m e s a d a y w hil st c o nfi n ed.  
g  H a e m at ol o g y a n d bi o c h e mi str y at s cr e e ni n g i n cl u d e s li pi d s f or c ar di o v a s c ul ar ri s k f a ct or.  
h  Ri a m et ®  will b e a d mi ni st er e d t o all p arti ci p a nt s o n D a y 4 7 ± 2 or e arli er a s o utli n e d i n S e cti o n 8. 1. 7. P arti ci p a nt s m a y b e tr e at e d wit h Pri m a ci n ® if g a m et o c yt a e mi a i s s u s p e ct e d fr o m 
p ar a sit e lif e c y c l e st a g e q R T- P C R at t h e I n v e sti g at or’ s di s cr eti o n, t o e n s ur e cl e ar a n c e of g a m et o c yt e s. If n e e d e d, p arti ci p a nt s will b e a d mi ni st er e d a si n gl e or al d o s e of si x Pri m a ci n ® 
t a bl et s (t ot al d o s e 4 5 m g pri m a q ui n e) wit h f o o d, u nl e s s G 6 P D d efi ci e nt i n w hi c h c a s e a l o w d o s e of 1 5 m g (t w o t a bl et s) will b e a d mi ni st er e d. 
I Bl o o d s a m pl e s f or m al ari a 1 8 S q P C R will b e c oll e ct e d f oll o wi n g p yr o n ari di n e tr e at m e nt at 4, 8, 1 2, 1 6, 2 0, 2 4, 3 0, 3 6, 4 2, 4 8, 6 0, 7 2, 8 4 a n d 9 6 h o ur s. A s o ut p ati e nt s, bl o o d will b e 
c oll e ct e d at l e a st t hr e e ti m e s p er w e e k u ntil a nti m al ari al r e s c u e tr e at m e nt. All o w e d ti m e wi n d o w s f or s a m pl e c oll e cti o n ar e s p e cifi e d i n S e cti o n 8. 4.  
j Bl o o d s a m pl e s f or dr u g c o n c e ntr ati o n m e a s ur e m e nt s will b e c oll e ct e d f oll o wi n g p yr o n ari di n e tr e at m e nt at 1, 2, 3, 4, 6, 8, 1 2, 1 6, 2 0, 2 4, 3 0, 3 6, 4 2, 4 8, 6 0, 7 2, 8 4 a n d 9 6 h o ur s. A s 
o ut p ati e nt s, bl o o d will b e c oll e ct e d o n t h e f oll o wi n g d a y s: 1 3, 1 5, 1 7, 2 0, 2 2, 2 4, 2 7, 2 9, 3 2, 3 9, 4 4, 4 7 a n d 5 0. All o w e d ti m e wi n d o w s f or s a m pl e c oll e cti o n ar e s p e cifi e d i n S e cti o n 
8. 4.  
k Bl o o d s a m pl e s f or p ar a sit e vi a bilit y e x vi v o gr o wt h will b e c oll e ct e d f oll o wi n g p yr o n ari di n e tr e at m e nt at 4, 8, 1 2, 1 6, 2 0, 2 4, 4 8, 7 2 a n d 9 6 h o ur s.  
l P erf or m S A R S -C o V - 2 t e sti n g at t h e eli gi bilit y vi sit ( N P A P C R), or at t h e s cr e e ni n g vi sit if t hi s vi sit i s c o n d u ct e d t w o d a y s pri or t o i n o c ul ati o n, D a y 0 ( R A T), D a y 6 a m ( N P A P C R) 
a n d D a y 8 ( R A T). F urt h er d et ail s ar e s p e cifi e d i n S e cti o n 8. 3. 7.  
m Bl o o d s a m pl e s f or f ut ur e m al ari a r e s e ar c h will b e c oll e ct e d d uri n g t h e o ut p ati e nt m o nit ori n g p h a s e o n D a y s 1 5 ± 1 a n d 2 9 ± 1 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 4  of 4 9  
 
3. 4  O bj e cti v e s  
3. 4. 1  Pri m ar y O bj e cti v e  
•  
T o c h ar a ct eri z e t h e P K/ P D r el ati o n s hi p of p yr o n ari di n e i n h e alt h y p arti ci p a nt s 
e x p eri m e nt all y i nf e ct e d wit h bl o o d st a g e P. f al ci p ar u m.  
3. 4. 2  S e c o n d ar y O bj e cti v e( s)  
•   T o e v al u at e t h e s af et y a n d t ol er a bilit y of si n gl e or al d o s e s of p yr o n ari di n e i n h e alt h y 
p arti ci p a nt s e x p eri m e nt all y i nf e ct e d wit h bl o o d st a g e P. f al ci p ar u m . 
•   T o c h ar a ct eri z e t h e p ar a sit e cl e ar a n c e ki n eti c s f oll o wi n g si n gl e d o s e s of p yr o n ari di n e 
i n h e alt h y p arti ci p a nt s e x p eri m e nt all y i nf e ct e d wit h bl o o d- st a g e P. f al ci p ar u m . 
•   
T o  c h ar a ct eri z e  t h e  p h ar m a c o ki n eti c s  of  p yr o n ari di n e  f oll o wi n g  si n gl e  or al  d o s e 
a d mi ni str ati o n  i n  h e alt h y  p arti ci p a nt s  e x p eri m e nt all y  i nf e ct e d  wit h  bl o o d  st a g e P. 
f al ci p ar u m. 
•   T o c h ar a ct eri z e t h e eff e ct of p yr o n ari di n e o n P. f al ci p ar u m  3 D 7 p ar a sit e vi a bilit y u si n g 
p h e n ot y pi c a n d m ol e c ul ar a p pr o a c h e s. 
 
3. 5  E n d p oi nt s  
3. 5. 1  Pri m ar y E n d p oi nt( s)  
•   
T h e  P K/ P D  r el ati o n s hi p b et w e e n  p yr o n ari di n e  bl o o d  c o n c e ntr ati o n s  a n d bl o o d  st a g e 
a s e x u al p ar a sit a e mi a. 
3. 5. 2  S e c o n d ar y E n d p oi nt ( s)  
3. 5. 2. 1  S af et y P ar a m et er s 
T h e  s af et y  a n d  t ol er a bilit y  of  p yr o n ari di n e  will  b e  d et er mi n e d  b y  r e c or di n g  t h e  i n ci d e n c e, 
s e v erit y  a n d  r el ati o n s hi p  t o  p yr o n ari di n e  of  a d v er s e  e v e nt s  ( a s  d et er mi n e d  b y  cli ni c al, 
l a b or at or y a n d E C G e x a mi n ati o n s) fr o m D a y 8 u p t o D a y 5 0 ± 2.:  
•   
A E s (i n cl u di n g S A E s a n d A E SI s) 
•   
Vit al si g n s 
•  
1 2 -l e a d E C G 
•  
H a e m at ol o g y, Bi o c h e mi str y a n d Uri n al y si s 
•   
P h y si c al e x a mi n ati o n 
 
3. 5. 2. 2  P ar a sit e Cl e ar a n c e Ki n eti c s 
•   
T h e p ar a sit e cl e ar a n c e ki n eti c s f oll o wi n g d o si n g wit h p yr o n ari di n e will b e d et er mi n e d b y 
c al c ul ati n g t h e p ar a sit e r e d u cti o n r ati o o v er a 4 8- h o ur p eri o d ( P R R 4 8) a n d c orr e s p o n di n g 
p ar a sit e cl e ar a n c e h alf- lif e ( P Ct1/ 2 ) 
 
3. 5. 2. 3  P K P ar a m et er s  
•  
T h e p h ar m a c o ki n eti c s of p yr o n ari di n e will b e d et er mi n e d b y c al c ul ati n g t h e f oll o wi n g 
p ar a m et er s u si n g n o n- c o m p art m e nt al m et h o d s:  


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 5  of 4 9  
 
P ar a m et er s: 
C m a x : M a xi m u m bl o o d  c o n c e ntr ati o n o b s er v e d 
tm a x : Ti m e t o r e a c h m a xi m u m bl o o d  c o n c e ntr ati o n 
A U C l a st: Ar e a u n d er t h e c ur v e fr o m ti m e 0 t o l a st m e a s ur a bl e c o n c e ntr ati o n 
A U C i nf: Ar e a u n d er t h e c ur v e fr o m ti m e 0 t o i nfi nit y 
t½ : Half -lif e 
C L/ F : A p p ar e nt Or al Cl e ar a n c e 
V d / F: Ap p ar e nt v ol u m e of di stri b uti o n  
λ z : T er mi n al r at e c o n st a nt 
 
3. 6  S a m pl e Si z e  
A m a xi m u m s a m pl e si z e of 1 8 p arti ci p a nt s i s e x p e ct e d t o b e s uffi ci e nt t o a c hi e v e t h e pri m ar y 
e n d p oi nt of t h e st u d y, w hi c h i s t o d et er mi n e t h e P K/ P D p ar a m et er s of p yr o n ari di n e. T hi s s a m pl e 
si z e i s n ot b a s e d o n f or m al st ati sti c al c al c ul ati o n s b ut r at h er i s b a s e d o n pr e vi o u s e x p eri e n c e 
of c h ar a ct eri zi n g t h e P K/ P D r el ati o n s hi p of a nti m al ari al s u si n g t h e I B S M st u d y tri al s (r ef er t o 
t h e  Bl o o d  st a g e  Pl a s m o di u m  f al ci p ar u m  c h all e n g e  a g e nt  P.  f al ci p ar u m  3 D 7  I n v e sti g at or’ s 
Br o c h ur e).  T h e  s p e cifi c  n u m b er  of  p arti ci p a nt s r e q uir e d  will  b e  d e pe n d e nt  o n  t h e  e m er gi n g 
r e s ult s d uri n g t h e st u d y; it i s p o s si bl e t h at l e s s t h a n 1 8 p arti ci p a nt s will b e r e q uir e d.  
 
3. 7  Tr e at m e nt A s si g n m e nt a n d R a n d o mi s ati o n  
All p arti ci p a nt s will r e c ei v e a n i d e ntifi c ati o n n u m b er ( s cr e e ni n g n u m b er) a s s o o n a s t h e y h a v e 
si g n e d t h e i nf or m e d c o n s e nt f or m. P arti ci p a nt s w h o m e et t h e eli gi bilit y crit eri a a n d ar e 
e nr oll e d i n t h e st u d y will b e a s si g n e d a n e nr ol m e nt n u m b er o n D a y 0 i m m e di at el y pri or t o 
a d mi ni str ati o n of t h e m al ari a c h all e n g e a g e nt. P arti ci p a nt s will b e r a n d o mi s e d wit hi n e a c h 
c o h ort t o a d o s e gr o u p o n D a y 8, i m m e di at el y pri or t o a d mi ni str ati o n of t h e I M P. T hi s i s a n 
o p e n l a b el st u d y a n d t h er ef or e n o bli n di n g will b e p erf o r m e d.  
R e pl a c e m e nt of Wit h dr a w n/ Di s c o nti n u e d P arti ci p a nt s  
P arti ci p a nt s w h o si g n t h e i nf or m e d c o n s e nt f or m, a n d ar e r a n d o mi s e d a n d r e c ei v e m al ari a 
i n o c ul ati o n, a n d s u b s e q u e ntl y wit h dr a w, or ar e wit h dr a w n or di s c o nti n u e d fr o m t h e tri al, m a y 
b e r e pl a c e d aft er  m ut u al a gr e e m e nt b et w e e n t h e S p o n s or a n d t h e PI. T h e d e ci si o n r e g ar di n g 
t h e r e pl a c e m e nt of p arti ci p a nt s will b e d o c u m e nt e d.  
 
P arti ci p a nt s w h o h a v e b e e n i n o c ul at e d a n d i n di c at e t h e y wi s h t o wit h dr a w fr o m t h e tri al m u st 
c o m pl et e t h e f ull c o ur s e of d efi niti v e a nti m al ari al tr e at m e nt. 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 6  of 4 9  
 
4.  S T A TI S TI C A L C O N SI D E R A TI O N S  
D at a a n al y si s will b e p erf or m e d a c c or di n g t o t h e S p o n s or’ s r e pr e s e nt ati v e St a n d ar d 
O p er ati n g Pr o c e d ur e s ( S O P s). 
 
T h e g e n er al a n al yti c al a p pr o a c h f or all e n d p oi nt s will b e d e s cri pti v e i n n at ur e.  All s u m m ari e s 
will pr e s e nt t h e d at a b y d o s e gr o u p a s w ell a s b y all p arti ci p a nt s c o m bi n e d.  
 
U nl e s s ot h er wi s e st at e d, t h e f oll o wi n g st ati sti c al a p pr o a c h e s will b e t a k e n: 
 
C o nti n u o u s v ari a bl e s:     
D e s cri pti v e st ati sti c s will i n cl u d e t h e n u m b er of n o n- mi s si n g 
v al u e s, m e a n, st a n d ar d d e vi ati o n ( S D), C V %, m e di a n, 
i nt er q u artil e r a n g e, mi ni m u m, a n d m a xi m u m. T h e mi ni m u m a n d 
m a xi m u m v al u e s will b e pr e s e nt e d t o t h e s a m e n u m b er of 
d e ci m al pl a c e s a s r e c or d e d i n t h e r a w d at a; m e a n a n d m e di a n 
will b e pr e s e nt e d t o 1 d e ci m al pl a c e m or e t h a n r a w d at a; a n d 
S D will b e pr e s e nt e d t o t w o m or e d e ci m al pl a c e s t h a n t h e r a w 
d at a.  T h e n u m b er of d e ci m al s f or l a b p ar a m et er s m a y b e 
r e d u c e d if t h e f ull pr e s e nt ati o n of t h e d at a i n cl u d e s m or e t h a n 4 
si g nifi c a nt fi g ur e s.  I n g e n er al, t h e m e a n a n d m e di a n will b e 
r e p ort e d wit h o n e d e ci m al pl a c e m or e t h a n t h e d at a, a n d t h e 
st a n d ar d d e vi ati o n will b e r e p ort e d wit h t w o d e ci m al pl a c e s 
m or e t h a n t h e d at a . 
 
C at e g ori c al v ari a bl e s:     
D e s cri pti v e st ati sti c s will i n cl u d e fr e q u e n c y c o u nt s a n d 
p er c e nt a g e s p er c at e g or y.  P er c e nt a g e s will b e r o u n d e d t o 
o n e  d e ci m al pl a c e, wit h t h e d e n o mi n at or b ei n g t h e n u m b er of 
p ati e nt s i n t h e r el e v a nt p o p ul ati o n. 
 
I m p ut ati o n:   
N o i m p ut ati o n will b e p erf or m e d f or mi s si n g d at a.   
 
C o nfi d e n c e i nt er v al s ( CI s):     CI s will b e t w o- si d e d a n d will u s e 9 5 % c o nfi d e n c e l e v el s.  A n y 
a n al y s e s r e q uiri n g si g nifi c a n c e t e sti n g will u s e a t w o- si d e d t e st 
at t h e 5 % si g nifi c a n c e l e v el. 
 
U n s c h e d ul e d Vi sit s  
U n s c h e d ul e d vi sit s t h at d o n ot h a v e l a b or at or y d at a o n t h e s a m e 
vi sit d at e will b e e x cl u d e d fr o m s u m m ar y t a bl e s. 
 
E arl y t er mi n ati o n vi sit  
A s s e s s m e nt s c o n d u ct e d at E arl y T er mi n ati o n will b e e x cl u d e d 
fr o m vi sit- b a s e d s u m m ar y t a bl e s. 
 
 
4. 1  D at a C a pt ur e  
4. 1. 1  D at a b a s e  
T h e pri m ar y m et h o d of d at a c oll e cti o n i s vi a t h e st u d y d at a b a s e, d e v el o p e d wit hi n t h e c h o s e n 
El e ctr o ni c D at a C a pt ur e ( E D C) pl atf or m, I B M Cli ni c al D e v el o p m e nt. T h e d at a b a s e h a s b e e n 
d e si g n e d b a s e d o n t h e fi n al pr ot o c ol, t h e s y st e m/ c or e c o nfi g ur ati o n, el e ctr o ni c C a s e R e p ort 
F or m ( e C R F) s p e cifi c ati o n s a n d/ or m o c k e C R F a n d c o n si st e n c y c h e c k s p e cifi c ati o n s. 
 
D at a will b e e nt er e d dir e ctl y i nt o t h e E D C s y st e m. Sit e - c oll e ct e d d at a will b e e nt er e d dir e ctl y 
fr o m s o ur c e n ot e s at t h e sit e a n d will b e v erifi e d b y Cli ni c al R e s e ar c h A s s o ci at e s ( C R A s) t o 
e n s ur e d at a i nt e grit y.  


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 7  of 4 9  
 
 
R ef er t o t h e D at a M a n a g e m e nt Pl a n f or f urt h er d et ail s.  
4. 1. 2.  T hir d P art y D at a  
4. 1. 2. 1  S af et y L a b or at or y 
C e ntr al s af et y l a b or at or y d at a will b e r e c ei v e d fr o m S ulli v a n Ni c ol ai d e s ( S N P) P at h ol o g y 
C e ntr al L a b or at or y a s s p e cifi e d i n t h e D at a Tr a n sf er S p e cifi c ati o n. At l e a st o n e tr a n sf er will 
b e d eli v er e d pri or t o d at a b a s e l o c k a n d r e c o n cil e d a g ai n st C R F d at a. F oll o wi n g s u c c e s sf ul 
r e c o n cili ati o n a n d r e s ol uti o n of a n y d at a i s s u e s, t h e d at a will b e i n c or p or at e d i nt o t h e E n d of 
St u d y a n al y si s. 
 
N o u nit c o n v er si o n of l a b or at or y d at a will b e p erf or m e d.  
 
4. 1. 2. 2  P K L a b or at or y  
P K s a m pl e s will b e a n al y s e d b y S wi s s Bi o Q u a nt C e ntr al ( S B Q) a n d P K p ar a m et er s will b e 
d eri v e d b y P h ar m a Ki n eti c.  Fi n al P K a s s a y d at a will b e tr a n sf err e d t o S S R, a s s p e cifi e d i n t h e 
S B Q D T S, f or i n c or p or ati o n i nt o t h e P C S D T M. 
 
4. 1. 2. 3  P D L a b or at or y  
All p ar a sit a e mi a d at a will b e g e n er at e d b y t h e Q u e e n sl a n d P a e di atri c I nf e cti o u s Di s e a s e s 
( Q PI D) l a b or at or y a n d c a pt ur e d el e ctr o ni c all y u si n g M A R S ( 1 8 S q P C R p ar a sit a e mi a d at a). All 
p ar a sit e lif e- st a g e d at a (i. e. g a m et o c y t e mi a a n d ri n g- st a g e p ar a sit e s) will b e g e n er at e d b y t h e 
Q PI D l a b or at or y a n d tr a n sf err e d el e ctr o ni c all y t o t h e Q u e e n sl a n d I n stit ut e of M e di c al 
R e s e ar c h ( QI M R) B er g h of er a n d  t hi s d at a will b e u s e d f or P K/ P D m o d elli n g. QI M R will 
pr o vi d e t h e ‘r a w’ P C R d at a  1 8 s a n d R T - P C R t o S S R a s p er D T S, a n d fi n al P D p ar a m et er s 
will b e i n t h e P R R r e p ort.  
 
4. 1. 2. 4  P K/ P D L a b or at or y  
P K/ P D a n al y si s will b e p erf or m e d b y M M V, P h ar m a c o m etri c s. S S R will b e r e q uir e d t o pr o vi d e 
t h e P K c o n c e ntr ati o n a n d  p ar a sit e mi a d at a , a s w ell a s a n y C R F d at a s et s r e q uir e d t o p erf or m 
t h e P K/ P D a n al y s e s ( e. g. d e m o gr a p hi c s, dr u g a d mi ni str ati o n d at a).   
 
 
 
 
 
 
 
4. 2  St ati sti c al Pr o gr a m mi n g  
4. 2. 1  Pr o gr a m mi n g S p e cifi c ati o n s  
4. 2. 1. 1    
S af et y A n al y si s 
Pr o gr a m mi n g s p e cifi c ati o n s will b e pr e p ar e d t o d et ail t h e S A S pr o gr a m mi n g of C DI S C 
( S D T M a n d A D a M) d at a s et s a n d li sti n g s, t a bl e s a n d fi g ur e s. 
 
4. 2. 1. 2    
P D A n al y si s 
D at a m a ni p ul ati o n a n d d at a a n al y s e s f or all P D  d at a will b e p erf or m e d u si n g R ( hi g h e st 
v er si o n a v ail a bl e). 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 8  of 4 9  
 
 
4. 2. 1. 3    
P K A n al y si s 
T h e e sti m ati o n of p h ar m a c o ki n eti c p ar a m et er s b y n o n- c o m p art m e nt al a n al y si s ( N C A) 
m et h o d s will b e p erf or m e d u si n g P h o e ni x Wi n N o nli n s oft w ar e ( v 8. 3 or a m or e r e c e nt v er si o n, 
C ert ar a U S A, I n c., U S A). 
 
4. 2. 1. 4    
P K P D A n al y si s 
All d at a pr o c e s si n g, a n al y si s, m o d el s et u p a n d m o d eli n g r e s ult a n al y si s i n cl u di n g g o o d n e s s-
of -fit pl ot s will b e p erf or m e d i n R 4. 1. 3 a n d I Q Rt o ol p a c k a g e 1. 1 0. 0 [I Q Rt o ol s]. 
N L M E m o d elli n g will b e p erf or m e d wit h M o n oli x 2 0 1 9 R 1 [ M L X 1 9] u si n g St o c h a sti c 
A p pr o xi m ati o n E x p e ct ati o n M a xi mi z ati o n ( S A E M) f or p ar a m et er e sti m ati o n. 
4. 2. 2  B a s eli n e  
B a s eli n e will b e d efi n e d a s :  
 
S af et y  
•   T h e l a st s c h e d ul e d o b s er v ati o n pri or t o t h e a d mi ni str ati o n. of t h e m al ari a c h all e n g e 
i n o c ul u m [I n o c ul ati o n B a s eli n e], 
•  
T h e  l a st s c h e d ul e d o b s er v ati o n pri or t o t h e fir st a d mi ni str ati o n of I M P [ Tr e at m e nt 
B a s eli n e].  
 
P D  
•   T h e l a st o b s er v ati o n pri or t o t h e a d mi ni str ati o n of t h e I M P f or P D d at a.  
 
P K  
•   T h e l a st o b s er v ati o n pri or t o t h e a d mi ni str ati o n of t h e I M P f or P K d at a. 
 
4. 2. 3   Ch a n g e fr o m B a s eli n e  
C h a n g e fr o m B a s eli n e will b e c al c ul at e d a s: 
 
! ℎ# $ % &  ( ) * +  , # - &./ $ & = (2 * -3 , # - &./ $ &  4 #. 5 & ) −, # - &./ $ &  4 #. 5 &  
4. 2. 3. 1    
 D a y s r el ati v e t o I n o c ul ati o n  
 
  8 # 9 - = (: - - & - - + & $3  8 #3 & −; $ * < 5. #3/ * $ : = +/ $/ -3 ) #3/ * $  8 #3 & ) + 1  
 
4. 2. 3. 2   
D a y s r el ati v e t o I M P a d mi ni str ati o n  
 
 8 # 9 - = (: - - & - - + & $3  8 #3 & −@/ ) -3  A3 5 = 9  8 ) 5 %  : = +/ $/ -3 ) #3/ * $  8 #3 & ) + 1  
4. 2. 4  I M P Tr e at m e nt Gr o u p s  
T a b ul ati o n s will s u m m ari s e d at a b y t h e f oll o wi n g I M P gr o u p s a n d t h e s e m a y v ar y f or e a c h 
c o h ort:  
P yr 3 6 0 m g 
P yr 5 4 0 m g 
P yr 7 2 0 m g 
O v er all 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 1 9  of 4 9  
 
5.  A N A L Y SI S S E T S  
 
T h e a n al y si s s et s t o b e u s e d f or t h e a n al y s e s will b e : F ull A n al y s e s set ( F A S),  I M P s et , P K s et , 
P D s et a n d P K/ P D s et .   
T h e  n u m b er  of p arti ci p a nt s  i n  e a c h  a n al y si s  s et  will  b e  s u m m ari s e d,  wit h  a  c orr e s p o n di n g 
li sti n g. 
5. 1  A n al y si s S et D e s cri pti o n s  
5. 1. 1  F ull A n al y si s S et  
T h e F ull A n al y si s S et ( F A S) will c o n si st of all e nr oll e d p arti ci p a nt s, i. e. p arti ci p a nt s t h at r e c ei v e d 
t h e m al ari a c h all e n g e a g e nt.  
5. 1. 2  I M P S et 
T h e I M P S et will i n cl u d e all e nr oll e d p arti ci p a nt s a d mi ni st er e d wit h t h e I M P. 
 
5. 1. 3  P K A n al y si s S et  
T h e P K a n al y si s s et will i n cl u d e all p arti ci p a nt s wit h at l e a st o n e a v ail a bl e v ali d (i. e. n ot fl a g g e d 
f or  e x cl u si o n)  P K  c o n c e ntr ati o n  m e a s ur e m e nt,  w h o  r e c ei v e d  a n y  st u d y  tr e at m e nt  a n d 
e x p eri e n c e d n o pr ot o c ol d e vi ati o n s wit h r el e v a nt i m p a ct o n P K d at a. 
 
5. 1. 4  P D A n al y si s S et  
T h e P D a n al y si s s e t will i n cl u d e all p arti ci p a nt s i n o c ul at e d wit h t h e m al ari a c h all e n g e a g e nt a n d 
w h o d e v el o p p ar a sit a e mi a d et e ct a bl e b y q P C R, r e c ei v e d a d o s e of p yr o n ari di n e, a n d h a v e n o 
pr ot o c ol d e vi ati o n s or ot h er cir c u m st a n c e s t h at w o ul d e x cl u d e t h e m fr o m t h e P D a n al y si s.   
5. 1. 5  P K/ P D A n al y si s S et  
T h e P K/ P D a n al y si s s et will i n cl u d e all p arti ci p a nt s fr o m t h e P K s et a n d all p arti ci p a nt s fr o m t h e 
P D s et. 
 
6.  P R O T O C O L  D E VI A TI O N S  
A n al y si s S et : F A S 
 
All  pr ot o c ol d e vi ati o n s will  b e  li st e d, gr o u p e d b y  p arti ci p a nt a n d  d o s e gr o u p 
T h e pr ot o c ol d e vi ati o n s u m m ar y t a bl e will c a pt ur e all d e vi ati o n s a s mi n or d e vi ati o n s a n d all 
vi ol ati o n s a s i m p ort a nt d e vi ati o n s. A f o ot n ot e will b e a d d e d t o t h e s u m m ar y t a bl e d o c u m e nti n g 
t h e c h a n g e. 
T h e  pr ot o c ol d e vi ati o n s u m m ar y t a bl e will  i n cl u d e: 
•  
T h e  t ot al n u m b er  of  mi n or pr ot o c ol  d e vi ati o n s 
•  
T h e  t ot al n u m b er  of i m p ort a nt  pr ot o c ol  d e vi ati o n s 
•  
T h e  n u m b er  of P arti ci p a nt s w h o  r e p ort e d at  l e a st o n e  mi n or pr ot o c ol d e vi ati o n  
•  
T h e  n u m b er  of P arti ci p a nt s w h o  r e p ort e d at  l e a st o n e  i m p ort a nt pr ot o c ol  d e vi ati o n s 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 0  of 4 9  
 
7.  P A R TI CI P A N T  DI S P O SI TI O N  
A n al y si s S et : F A S 
Di s p o siti o n  
A li sti n g of p arti ci p a nt di s p o siti o n will pr e s e nt:  
•   
D at e of i nf or m e d c o n s e nt 
•  
D at e of i n o c ul ati o n  
•   
D at e of I M P a d mi ni str ati o n 
•   
D at e of r e s c u e m e di c ati o n a d mi ni str ati o n i n cl u di n g Pri m a ci n 
•   
Di d t h e p arti ci p a nt c o m pl et e t h e st u d y ? 
•   
D at e of c o m pl eti o n / e arl y wit h dr a w al 
•   
Pri m ar y r e a s o n f or e arl y wit h dr a w al (i n cl u di n g i n st a n c e s w h er e e arl y t er mi n ati o n w a s 
r el at e d t o C O VI D-1 9)  
D et ail e d e arl y  wit h dr a w al i nf or m ati o n  will al s o b e li st e d i n a s e p ar at e li sti n g. 
If t h er e ar e a n y d e at h s r e p ort e d, a s e p ar at e d e at h li sti n g will b e pr e p ar e d. 
T h e  n u m b er  a n d  p er c e nt a g e  of  P arti ci p a nt s  e nt eri n g  a n d  di s c o nti n ui n g  t h e  st u d y  will  b e 
s u m m ari s e d  b y d o s e  gr o u p a n d  o v er all  al o n g  wit h  t h e  r e a s o n  f or  di s c o nti n u ati o n.  T h e 
p arti ci p a nt di s p o siti o n s u m m ar y t a bl e will i n cl u d e: 
 
•  
N u m b er of p arti ci p a nt s si g n e d i nf or m e d c o n s e nt 
•  
N u m b er of p arti ci p a nt s w h o r e c ei v e d I M P  
•  
N u m b er of p arti ci p a nt s w h o c o m pl et e d t h e f ull st u d y 
•  
N u m b er of p arti ci p a nt s wit h dr a w n fr o m t h e st u d y e arl y 
•   
R e a s o n f or e arl y wit h dr a w al 
8.  D E M O G R A P HI C  A N D B A S E LI N E I N F O R M A TI O N  
A n al y si s S et : F A S  
8. 1   
D e m o gr a p hi c s  
D e m o gr a p hi c d at a will b e li st e d f or all e nr oll e d p arti ci p a nt s a n d s u m m ari s e d b y d o s e gr o u p 
a n d o v er all.  D at a i n cl u d e s: 
A g e  
S e x  
W o m e n of C hil d -b e ari n g p ot e nti al  
P o st- m e n o p a u s al   
R a c e 
Et h ni cit y 
W ei g ht ( k g) 
H ei g ht ( m)  
B MI ( k g/ m 2) 
8. 2  M e di c al Hi st or y  
All m e di c al hi st or y d at a will b e li st e d, gr o u p e d b y p arti ci p a nt. 
 
M e di c al hi st or y will b e c o d e d u si n g t h e M e di c al Di cti o n ar y f or R e g ul at or y A cti viti e s ( M e d D R A   
v er si o n 2 4. 1) a n d s u m m ari s e d b y s y st e m or g a n cl a s s ( S O C) a n d pr ef err e d t er m ( P T).  
8. 3  Pri or M e di c ati o n s  
Pri or m e di c ati o n s ar e d efi n e d a s a n y m e di c ati o n t h at i s st art e d b ef or e fir st i n o c ul ati o n wit h t h e 
m al ari a c h all e n g e a g e nt , r e g ar dl e s s of w h e n it e n d e d. If a m e di c ati o n h a s a mi s si n g or p arti al 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 1  of 4 9  
 
mi s si n g st art/ e n d d at e or ti m e a n d it c a n n ot b e d et er mi n e d w h et h er it w a s t a k e n b ef or e I M P 
or c o n c o mit a ntl y, it will b e c o n si d er e d a s pri or a n d c o n c o mit a nt. M e di c ati o n s will b e c o d e d 
u si n g W H O Dr u g Di cti o n ar y S e pt e m b er 2 0 2 1 r el e a s e. 
 
Pri or m e di c ati o n s will b e s u m m ari s e d b y A n at o mi c al T h er a p e uti c Cl a s s ( A T C) a n d pr ef err e d 
n a m e ( P N) a n d will al s o b e li st e d. T h e s u m m ar y t a bl e s will s h o w t h e n u m b er a n d p er c e nt a g e 
of p arti ci p a nt s t a ki n g e a c h m e di c ati o n b y A T C a n d P N. 
8. 4  Dr u g S cr e e n  
Dr u g s cr e e n d at a will b e li st e d f or all p arti ci p a nt s. 
8. 5  Al c o h ol S cr e e n  
Al c o h ol s cr e e ni n g d at a will b e li st e d f or all p arti ci p a nt s. 
8. 6  S A R S -C o V -2 S cr e e ni n g  
A n y p o siti v e S A R S- C o V 2 s cr e e ni n g r e s ult s will b e li st e d. 
8. 7  F olli cl e Sti m ul ati n g H or m o n e ( F S H)  
F S H r e s ult s will b e li st e d f or P o st- m e n o p a u s al w o m e n o nl y. 
8. 8  B e c k D e pr e s si o n I n v e nt or y  
B e c k D e pr e s si o n I n v e nt or y ( B DI-II) d at a will b e li st e d f or all p arti ci p a nt s, i n cl u di n g a n s w er s t o 
t h e i n di vi d u al q u e sti o n s a s w ell a s t h e t ot al s c or e s, w hi c h ar e a ut o- c al c ul at e d wit hi n t h e C R F. 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 2  of 4 9  
 
9.  T R E A T M E N T  E X P O S U R E  
A n al y si s S et : F A S S et  
 
P arti ci p a nt e x p o s ur e t o t h e pr ot o c ol- s p e cifi e d tr e at m e nt s (I M P, m al ari a c h all e n g e a g e nt a n d 
r e s c u e m e di c ati o n s Ri a m et, M al ar o n e , I V Art e s u n at e a n d Pri m a ci n) will b e li st e d a n d 
s u m m ari s e d b y  d o s e of I M P a n d o v er all.   
 
1 0.   PRI M A R Y E N D P OI N T A N A L Y SI S  (P K/ P D A N A L Y SI S)  
 
A n al y si s S et: P K/ P D A n al y si s S et  
 
1 0. 1  
O v er vi e w of m et h o d ol o g y f or P K/ P D M o d elli n g  
A p o p ul ati o n P K P D m o d el will b e d eri v e d fr o m t h e c o n c e ntr ati o n s a n d p ar a sit e mi a o b s er v e d i n 
t h e c h all e n g e p arti ci p a nt s. It i s a c hi e v e d b y s e q u e nti all y d e v el o pi n g a p o p ul ati o n P K m o d el t o 
d e s cri b e  t h e o b s er v e d  i n di vi d u al  P K  pr ofil e s a n d  a  p o p ul ati o n P D  m o d el  f or  q u a ntif yi n g  t h e 
r el ati o n s hi p of bl o o d c o n c e ntr ati o n s a n d t h e p ar a sit e cl e ar a n c e. 
T h e n, t h e P D m o d el i s c o m bi n e d wit h a P K m o d el i n m al ari a p ati e nt s t o pr e di ct t h e p ar a sit e mi a 
pr ofil e s t hi s p o p ul ati o n, a s s u mi n g t h e P K P D r el ati o n s hi p will b e t h e s a m e i n p ati e nt s a s i n t h e 
c h all e n g e p arti ci p a nt s . 
1 0. 2  
S o ur c e s of D at a  
D at a fr o m st u d y M M V _ P yr o n ari di n e _ 2 1 _ 0 1 will c o nt ai n i nf or m ati o n o n t h e i n di vi d u al 
P yr o n ari di n e t etr a p h o s p h at e d o s e s, P yr o n ari di n e* P K c o n c e ntr ati o n s, t h e tri pli c at e d 
p ar a sit e mi a l e v el s, g a m et o c yt e d at a a n d t y pi c al c o v ari at e s s u c h a s a g e, h ei g ht, w ei g ht, s e x 
a n d r a c e  
 
1 0. 3  
P h ar m a c o ki ni eti c s  
Bl o o d f or P yr o n ari di n e c o n c e ntr ati o n s will b e c oll e ct e d pr e- P yr o n ari di n e t etr a p h o s p h at e 
a d mi ni str ati o n o n D a y 8, t h e n at t h e f oll o wi n g h o ur s p o st- P yr o n ari di n e t etr a p h o s p h at e 
a d mi ni str ati o n w hil e c o nfi n e d i n t h e cli ni c al u nit: 1, 2, 3, 4, 6, 8, 1 2, 1 6, 2 0, 2 4, 3 0, 3 6, 4 2, 4 8, 
6 0, 7 2, 8 4 a n d 9 6 h o ur s. A s o ut p ati e nt s, p arti ci p a nt s will h a v e bl o o d c oll e ct e d o n t h e f oll o wi n g 
d a y s: 1 3, 1 5, 1 7, 2 0, 2 2, 2 4, 2 7, 2 9, 3 2, 3 9, 4 4, 4 7 a n d 5 0. 
1 0. 4  
P h ar m a c o d y n a mi c s  
1 0. 4 . 1 M al ari a 1 8 S q P C R  
Bl o o d will b e c oll e ct e d t o q u a ntif y t ot al m al ari a p ar a sit e mi a b y q u a ntit ati v e p ol y m er a s e c h ai n 
r e a cti o n t ar g eti n g t h e g e n e e n c o di n g P. f al ci p ar u m 1 8 S r R N A ( m al ari a 1 8 S q P C R; r e p ort e d 
a s p ar a sit e s/ 5 0 0 u L p a c k e d R B C s). M al ari a 1 8 S q P C R d at a will b e u s e d t o c al c ul at e t h e t ot al 
p ar a sit e c o u nt s. Bl o o d f or m al ari a 1 8 S q P C R will b e c oll e ct e d o n D a y 0 pri or t o i n o c ul ati o n 
wit h t h e m al ari a c h all e n g e a g e nt, t h e n o n c e d ail y o n D a y s 4, 5, 6 a n d 7. O n D a y 8, bl o o d will 
b e c oll e ct e d pr e- P yr o n ari di n e t etr a p h o s p h at e a d mi ni str ati o n, t h e n at t h e f oll o wi n g h o ur s p o st-
P yr o n ari di n e t etr a p h o s p h at e a d mi ni str ati o n w hil e c o nfi n e d i n t h e cli ni c al u nit: 4, 8, 1 2, 1 6, 2 0, 
2 4, 3 0, 3 6, 4 2, 4 8, 6 0, 7 2, 8 4 a n d 9 6 h o ur s. A s o ut p ati e nt s, p arti ci p a nt s will h a v e bl o o d 
c oll e ct e d at l e a st t hr e e ti m e s p er w e e k u ntil a nti m al ari al r e s c u e tr e at m e nt. 
1 0. 4. 2  P ar a sit e lif e c y cl e st a g e q R T -P C R  


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 3  of 4 9  
 
A d diti o n al bl o o d s a m pl e s m a y b e c oll e ct e d t o e v al u at e t h e pr e s e n c e of s e x u al p ar a sit e 
st a g e s ( g a m et o c yt e s) a n d ot h er p ar a sit e lif e c y cl e st a g e s u si n g q u a ntit ati v e r e v er s e-
tr a n s cri pt a s e p ol y m er a s e c h ai n r e a cti o n ( q R T-P C R). T h e q R T - P C R a s s a y s will b e d o n e f or 
t h e f oll o wi n g t ar g et s: t h e f e m al e g a m et o c yt e- s p e cifi c tr a n s cri pt pf s 2 5, t h e m al e g a m et o c yt e-
s p e cifi c tr a n s cri pt Pf M G E T, a n d t h e ri n g- st a g e tr a n s cri pt pf S B P- 1.  
 
1 0. 5 
D at a A s s e m bl y  
Fir st a m a st er d at a s et will b e a s s e m bl e d f or m att e d a c c or di n g t o t h e M M V d at a s et st a n d ar d. 
( cf., " M S _I Q d at a s et _ V 4 _ 2 0 2 1 0 2 1 9. d o c x"). Bl o o d  c o n c e ntr ati o n will b e i n u g/ m L, t ot al p ar a sit e 
l e v el s i n c o u nt s/ m L, a n d d o s e s i n m g. G a m et o c yt e d at a i s i n c o pi e s/ u L.  
P ar a sit e d at a aft er t h e fir st a d mi ni str ati o n of r e s c u e m e di c ati o n i s fl a g g e d b y s etti n g t h e 
I G N O R E c ol u m n t o " R e s c u e d". T h e ti m e i n h o ur s i s r el ati v e t o t h e (fir st) P yr o n ari di n e 
t etr a p h o s p h at e a d mi ni str ati o n ti m e. 
A g e n er al a n al y si s d at a s et i s d eri v e d fr o m t h e m a st er d at a s et. G a m et o c yt e d at a will b e 
c o n v ert e d t o p ar a sit e c o u nt s f oll o wi n g t h e i n str u cti o n s i n " A n al y zi n g R N A d at a b y K. C olli n s 
N o v 2 0 1 7. d o c x". T ot al c o u nt s of p ar a sit e s w er e fl a g g e d t o b e i g n or e d ( b y s etti n g t h e c ol u m n 
I G N O R E t o " g a m et o c yt e s") if c o u nt s of g a m et o c yt e s e x c e e d e d 1 0 % of t ot al c o u nt s. 
T h e vi a bl e p ar a sit e s c o u nt s will b e d eri v e d b y m ulti pli c ati o n of t h e vi a bl e fr a cti o n v al u e s wit h 
t h e m e a n p ar a sit e c o u nt at tr e at m e nt a d mi ni str ati o n. 
A g e n er al a n al y si s d at a s et i s d eri v e d fr o m t h e m a st er d at a s et. T h e g e n er al a n al y si s d at a s et 
will c o nt ai n t h e f oll o wi n g o b s er v ati o n s: P yr o n ari di n e bl o o d c o n c e ntr ati o n s, t ot al c o u nt s of 
p ar a sit e s, a s w ell a s c o u nt s of g a m et o c yt e s a n d vi a bl e p ar a sit e c o u nt s. I n a d diti o n, d o si n g 
r e c or d s f or P. f al ci p ar u m i n o c ul ati o n, P yr o n ari di n e t etr a p h o s p h at e a d mi ni str ati o n a n d r e s c u e 
m e di c ati o n a d mi ni str ati o n will b e d efi n e d. It i s a d vi s e d t o d efi n e a d diti o n al c ol u m n s 
D O S E L E V E L a n d D O S E M U L T t o i n di c at e t h e d o s e a m o u nt a n d n u m b er of d o s e s a 
p arti ci p a nt r e c ei v e d a s c o v ari at e c ol u m n s. 
N L M E d at a s et s f or t h e P K a n d P D m o d eli n g will b e d eri v e d fr o m t h e g e n er al d at a s et. 
 
1 0. 5. 1  P K m o d eli n g d at a s et  
O b s er v ati o n s b el o w t h e l o w er li mit of q u a ntifi c ati o n ( L L O Q) will b e c e n s or e d b y s etti n g t h e 
C E N S c o l u m n f or t h e s e o b s er v ati o n r e c or d s t o 1 a n d s etti n g t h e c orr e s p o n di n g d e p e n d e nt 
v ari a bl e ( D V) c ol u m n t o t h e L L O Q ( → M 3 m et h o d). O b s er v ati o n r e c or d s wit h mi s si n g v al u e s 
a n d r e c or d s wit h mi s si n g ti m e s will b e i g n or e d. R e c or d s fl a g g e d t o b e i g n or e d will b e 
r em o v e d fr o m t h e d at a s et. O b s er v ati o n s n ot a n al y s e d i n t h e P K m o d elli n g (i. e., all p ar a sit e 
c o u nt r e c or d s) will b e r e m o v e d. A s a r e s ult, t h e P K m o d elli n g d at a s et o nl y c o nt ai n s 
P yr o n ari di n e t etr a p h o s p h at e d o si n g r e c or d s a n d P yr o n ari di n e bl o o d c o n c e ntr ati o n r e c or d s. 
 
 
1 0. 5. 2  P D m o d eli n g d at a s et  
O b s er v ati o n s b el o w t h e L L O Q will b e c e n s or e d b y s etti n g t h e C E N S c ol u m n f or t h e s e 
o b s er v ati o n r e c or d s t o 1 a n d t h e c orr e s p o n di n g D V v al u e t o t h e L L O Q, i. e., 1 0 p/ m L ( → M 3 
m et h o d). T ot al c o u nt s of p ar a sit e s f or w hi c h c o u nt s of g a m et o c yt e s e x c e e d e d 1 0 % of t ot al 
c o u nt s ar e fl a g g e d t o b e i g n or e d ( b y s etti n g t h e c ol u m n I G N O R E t o " g a m et o c yt e s"). 
O b s er v ati o n r e c or d s wit h mi s si n g v al u e s a n d r e c or d s wit h mi s si n g ti m e s will b e i g n or e d. 
R e c or d s fl a g g e d t o b e i g n or e d will b e r e m o v e d fr o m t h e d at a s et. O b s er v ati o n s n ot a n al y s e d 
i n t h e P D m o d elli n g a n al y si s (i. e., c o n c e ntr ati o n r e c or d s) will b e r e m o v e d. A s a r e s ult, t h e P D 
m o d elli n g d at a s et o nl y c o nt ai n s P yr o n ari di n e t etr a p h o s p h at e d o si n g r e c or d s, t ot al p ar a sit e 
c o u nt s (i. e., tri pli c at e s). 
T h e m o d el b a s eli n e p ar a sit e mi a v al u e i s a d d e d t o t h e d at a s et a s g e o m etri c m e a n of p ar a sit e 
c o u nt s at ti m e of fir st P yr o n ari di n e t etr a p h o s p h at e d o s e. 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 4  of 4 9  
 
T h e t ot al p ar a sit e c o u nt s will b e l o g-tr a n sf or m e d f or t h e P D m o d elli n g a n d e sti m at e d 
i n di vi d u al P K m o d el p ar a m et er s will b e a d d e d t o t h e d at a s et a s r e gr e s s or s. 
 
 
1 0. 6.  M o d el b uil di n g  
1 0. 6 . 1.  
G e n er al P K P D m o d el b uil di n g a p pr o a c h  
P o p ul ati o n m o d el s of t h e P K a n d P D will b e d e v el o p e d i n st e p- wi s e m a n n er. 
1.  
A  p o p ul ati o n  P K  m o d el  will  b e  d e v el o p e d  t o  o bt ai n i n di vi d u al  P K  p ar a m et er  e sti m at e s 
b a s e d o n w hi c h t h e i n di vi d u al P K pr ofil e s ar e d e s cri b e d w ell. 
2.   
A p ar a sit e gr o wt h m o d el ( p ar a sit e gr o wt h r at e a n d d e vi ati o n of p ar a sit e mi a at i n o c ul ati o n 
fr o m  p ar a sit e mi a  at  tr e at m e nt  a d mi ni str ati o n)  will  b e  e sti m at e d  fr o m  t h e  t ot al  p ar a sit e 
c o u nt s b ef or e tr e at m e nt a d mi ni str ati o n. E sti m ati o n of t y pi c al a n d i n di vi d u al P D p ar a m et er s 
will  b e  p erf or m e d  wit h  I Q Rt o ol s  S y s Fit.  Ot h er  a p pr o pri at e  gr o wt h  m o d el s,  e. g.  a n 
o s cill at or y m o d el, will b e t e st e d if a d vi s a bl e. 
3.  
T h e P K P D m o d el wil l b e b uil d u si n g t h e i n di vi d u al P K p ar a m et er e sti m at e s a n d i n di vi d u al 
gr o wt h m o d el p ar a m et er e sti m at e s a s r e gr e s si o n p ar a m et er s. E sti m ati o n of t y pi c al a n d 
i n di vi d u al P D p ar a m et er s will fir st b e p erf or m e d wit h I Q Rt o ol s S y s Fit t o i n v e sti g at e m o d el 
i d e ntifi a bilit y a n d s e c o n d wit h M o n oli x t o a d diti o n all y e sti m at e II V. 
E sti m ati o n s will b e d o n e u si n g gr a di e nt- b a s e d S y s Fit al g orit h m i n I Q Rt o ol s or S A E M 
al g orit h m i n M o n oli x. Ot h er a p pr o pri at e m et h o d s m a y b e u s e d if a d vi s a bl e. 
I nt er-i n di vi d u al v ari a bilit y (II V) i s i m pl e m e nt e d f or n or m all y di stri b ut e d p ar a m et er s u si n g t h e 
f oll o wi n g e q u ati o n wit h B C  a s p o p ul ati o n a v er a g e p ar a m et er, B D a s i n di vi d u al p ar a m et er, a n d 
E D a s r a n d o m eff e ct t h at i s di stri b ut e d n or m all y ar o u n d z er o (E D ∼G (0 ,J K )). 
B D = B C + E D 
T h e f oll o wi n g e q u ati o n i s u s e d f or l o g- n or m all y di stri b ut e d p ar a m et er s. 
l n(B D) = l n(B C ) + E D 
C o nti n u o u s c o v ari at e s o n l o g- n or m all y di stri b ut e d p ar a m et er s will b e i m pl e m e nt e d a s f oll o w s. 
B D = B C ⋅
! O P D
! O P Q R SD T U
V
⋅E D 
B D r e pr e s e nt s t h e i n di vi d u al p ara m et er, B C  t h e p o p ul ati o n m e a n v al u e, ! O P D t h e i n di vi d u al 
c o v ari at e v al u e, ! O P Q R SD T U , t h e p o p ul ati o n m e di a n c o v ari at e v al u e, W  t h e c o v ari at e 
c o effi ci e nt, a n d E D t h e i n di vi d u al r a n d o m eff e ct. 
C at e g ori c al c o v ari at e s o n l o g- n or m all y di stri b ut e d p ar a m et er s will b e i m pl e m e nt e d a s f oll o w s. 
B D = B C ⋅X Y Z & V [ ⋅\ ] ^ _ [` ⋅E D 
T h e i n di vi d u al c o v ari at e v al u e s ! O P Da ar e 1 if p arti ci p a nt i b el o n g s t o t h e jt h c at e g or y of t h e 
c o v ati at e a n d 0 ot h er wi s e. , &3 # Y  i s t h e c o v ari at e v al u e f or t h e jt h c at e g or y. F or o n e c at e g or y, 
i. e. t h e r ef er e n c e, , &3 # Y  i s 0. 
F or t h e N L M E m o d eli n g wit h M o n oli x, t h e l o g-li k eli h o o d a n d t h e Fi s h er i nf or m ati o n m atri x will 
b e a p pr o xi m at e d b y li n e ari z ati o n. T h e n u m b er of it er ati o n s i n t h e b ur n- i n a n d t h e 
a c c u m ul ati o n p h a s e will b e 5 0 0 a n d 2 0 0 r e s p e cti v el y, b ut mi g ht b e a dj u st e d if r e q uir e d b a s e d 
o n i n s p e cti o n of p ar a m et er e sti m at e a n d o bj e cti v e f u n cti o n tr a c e s al o n g e sti m ati o n it er ati o n s. 
I n di vi d u al p ar a m et er s will b e d et er mi n e d a s c o n diti o n al m o d e s. 
F or P D m o d elli n g wit h I Q Rt o ol s S y s Fit i m pl e m e nt ati o n a n o n- li n e ar fi x e d- eff e ct s m o d elli n g 
a p pr o a c h i s e m pl o y e d. T y pi c al a n d i n di vi d u al p ar a m et er s ar e b ot h tr e at e d a s fi x e d eff e ct s. 
T h e p ar a m et eri z ati o n of i n di vi d u al p ar a m et er s a n d c o v ari at e s i s f ull y c o n si st e nt wit h t h e 
N L M E p ar a m et eri z ati o n a b o v e. T h e v ari a n c e s J K  of N L M E r a n d o m eff e ct s ar e a c c o u nt e d f or 
b y q u a dr ati c pri or s, .b cD d c (E ) =
e f g
K h g , a d d e d t o t h e l o g-li k eli h o o d f u n cti o n, w h er e t h e v al u e s f or 
J K  ar e fi x e d a n d t h e E  p ar a m et er s ar e e sti m at e d. T h e pr ofil e-li keli h o o d m et h o d [ R A U 0 9 , 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 5  of 4 9  
 
K A S 1 9 ] i s u s e d t o c o m p ut e c o nfi d e n c e i nt er v al s of t h e p ar a m et er s e sti m at e d wit h t h e S y s Fit 
a p pr o a c h.  
 
 
 
1 0. 6 . 1. 1 
M o d el e v al u ati o n  
A s s e s s m e nt of m o d el a d e q u a c y a n d d e ci si o n s a b o ut i n cr e a si n g m o d el c o m pl e xit y will b e 
dri v e n b y t h e d at a a n d g ui d e d b y g o o d n e s s- of -fit crit eri a, i n cl u di n g 
1.   
vi s u al  i n s p e cti o n  of  di a g n o sti c  s c att er  pl ot s  ( o b s er v e d  v s.  pr e di ct e d  c o n c e ntr ati o n, 
r e si d u al/ w ei g ht e d r e si d u al v s. pr e di ct e d c o n c e ntr ati o n or ti m e a n d hi st o gr a m s of i n di vi d u al 
r a n d o m eff e ct s, f or e x a m pl e), 
2.   
s u c c e s sf ul  c o n v er g e n c e  of  t h e  mi ni mi z ati o n  r o uti n e  wit h  at  l e a st  2  si g nifi c a nt  di git s  i n 
p ar a m et er e sti m at e s, 
3.   
pl a u si bilit y of p ar a m et er e sti m at e s, 
4.   
pr e ci si o n of p ar a m et er e sti m at e s, 
5.   
c orr el ati o n b et w e e n m o d el p ar a m et er e sti m ati o n err or s < 0. 9 5, a n d 
6.   
t h e B a y e si a n i nf or m ati o n crit eri o n ( BI C), gi v e n t h e mi ni m u m o bj e cti v e f u n cti o n v al u e a n d 
n u m b er of e sti m at e d p ar a m et er s. 
F or t h e n o nli n e ar fi x e d eff e ct s m o d el s, a d diti o n al di a g n o sti c pl ot s will b e pr o d u c e d:  
7.   
o bj e cti v e f u n cti o n v al u e s a cr o s s m ulti pl e fit s fr o m r a n d o mi s e d i niti al g u e s s e s, a n d 
8.  
pr ofil e li k eli h o o d pl ot s  
All p ar a m et er e sti m at e s will b e r e p ort e d wit h a m e a s ur e of e sti m ati o n u n c ert ai nt y, s u c h a s t h e 
r el ati v e st a n d ar d err or of t h e e sti m at e s. T h e i n di vi d u al P K fit s will b e u s e d t o e v al u at e t h e 
a d e q u a c y t o u s e t h e i n di vi d u al P K p ar a m et er e sti m at e s a s r e gr e s si o n p ar a m et er s f or P D 
m o d eli n g.  
T h e a d e q u a c y of si m ul ati o n wit h t h e s el e ct e d m o d el s will b e e v al u at e d b y vi s u al pr e di cti v e 
c h e c k s ( V P C s). V P C s f or e a c h d o s e l e v el will b e p erf or m e d b y si m ul ati o n of t h e st u d y 2 0 0 
ti m e s t a ki n g p ar a m et er e sti m ati o n u n c ert ai nt y i nt o a c c o u nt. 9 5 %- c o nfi d e n c e i nt er v al s f or t h e 
5t h p er c e ntil e, t h e m e di a n, a n d t h e 9 5t h p er c e ntil e will b e d eri v e d a n d c o m p ar e d t o t h e 
c orr e s p o n di n g v al u e s b a s e d o n t h e d at a. 
1 0. 6 . 2.  
P K m o d eli n g  
T h e P K of P yr o n ari di n e will b e m o d el e d u si n g c o m p art m e nt al m o d el s d e s cri bi n g t h e 
a b s or pti o n aft er or al a d mi ni str ati o n, di stri b uti o n b et w e e n c e ntr al a n d p eri p h er al 
c o m p art m e nt s, a n d t h e eli mi n ati o n fr o m t h e c e ntr al c o m p art m e nt. T h e vi s u al d at a a n al y si s 
will g ui d e t h e s el e cti o n of m o d el s t h at will b e t e st e d ( e. g., wit h r e s p e ct t o n u m b er of 
c o m p art m e nt s, li n e ar or s at ur a bl e eli mi n ati o n, a b s or pti o n ki n eti c s a n d err or m o d el). 
C o v ari at e s will b e i n cl u d e d if a p pr o pri at e. E sti m ati o n s will b e d o n e u si n g S A E M al g orit h m i n 
M o n oli x. Ot h er a p pr o pri at e m et h o d s m a y b e u s e d if a d vi s a bl e. 
1 0. 6 . 3.  
P D m o d eli n g  
P D m o d el d e v el o p m e nt will b e d o n e u si n g gr a di e nt b a s e d al g orit h m s i n I Q R T o ol s S y s Fit. T h e 
s el e ct e d m o d el will t h e n b e e sti m at e d wit h S A E M i n M o n oli x t o all o w f or e sti m ati o n of II V. 
Ot h er a p pr o pri at e m et h o d s m a y b e u s e d if a d vi s a bl e. A s a g e n er al c o n c e pt t h e c h a n g e s i n 
li vi n g p ar a sit e i  ar e m o d el e d a s t h e eff e ct of a n et e x p o n e nti al gr o wt h r at e j k  a n d a killi n g or 
cl e ar a n c e r at e l/..  d u e t o P yr o n ari di n e. T h e i niti al p ar a sit e mi a i m T n R  at ti m e 3C  will b e d eri v e d 
fr o m t h e m e a n p ar a sit e mi a w hi c h w a s o b s er v e d at tr e at m e nt a d mi ni str ati o n a n d a p ar a m et er 
i o & ) )  t o all o w f or p o p ul ati o n a n d i n di vi d u al d e vi ati o n s. T h e e q u ati o n s ar e e x pr e s s e d i n t h e 
l o g- s c al e s u c h a s: 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 6  of 4 9  
 
= i o
=3
=
j k −l/..
i o (3C )
=
i o m T n R + i o & ) )
 
w h er e i o = l n(i ) i s t h e l o g-tr a n sf or m e d p ar a sit e c o u nt s. 
  
T hr e e diff er e nt m o d el s d e s cri bi n g t h e r el ati o n s hi p of P yr o n ari di n e bl o o d c o n c e ntr ati o n o n t h e 
killi n g r at e will b e t e st e d: 
  
( 1) T h e " p Q T q - m o d el" a s s u m e s a dir e ct eff e ct of c o n c e ntr ati o n s o n p ar a sit e killi n g/ cl e ar a n c e, 
s u c h 
a s: 
l/.. = p Q T q ⋅
! rsDtt
! r
sDtt + p ! u C
sDtt  
w h er e ! r  i s t h e c o n c e ntr ati o n i n t h e c e ntr al c o m p art m e nt, p Q T q  i s t h e m a xi m u m eff e ct of 
t h e dr u g, p ! u C  i s t h e c o n c e ntr ati o n t h at r e s ult s i n 5 0 % of t h e m a xi m u m eff e ct a n d v/..  i s 
t h e Hill c o effi ci e nt. 
( 2)  T h e " T ur n o v er m o d el" i m pl e m e nt s a n i n dir e ct r e s p o n s e m o d el r efl e cti n g t h e i d e a t h at t h e 
ki n eti c s of dr u g-r e c e pt or bi n di n g l e a d t o a l a g g e d c o n c e ntr ati o n- eff e ct r el ati o n s hi p, s u c h 
a s: 
l/..
=
p Q T q ⋅k
= k
=3
=
w D U x
! rsDtt
! r
sDtt + p ! u C
sDtt −k y  
w h er e w D U r e pr e s e nt s t h e bi n di n g r at e of t h e dr u g t o a "r e c e pt or", w hi c h r e s ult s i n a l a g-
ti m e i n t h e r e s p o n s e. F urt h er m or e, p ! u C  r e pr e s e nt s t h e c o n c e ntr ati o n t h at r e s ult s i n a 5 0 % 
o c c u p a n c y of t h e r e c e pt or w hi c h l e a d s t o 5 0 % of t h e m a xi m u m eff e ct gi v e n e n o u g h ti m e 
t o r e a c h it. 
( 3) T h e " Cl e ar a n c e m o d el" a s s u m e s t h at t h e o b s er v e d p ar a sit e s ar e a mi xt ur e of li vi n g a n d 
d e a d p ar a sit e s. T h e dr u g h a s a dir e ct eff e ct o n t h e killi n g r at e of t h e p ar a sit e s, a n d t h e n 
t h e p ar a sit e s ar e cl e ar e d, b ut still o b s er v e d. T h er ef or e, t h e e q u ati o n s, i n t h e n or m al s c al e, 
ar e:  
= i ztD { R
=3
=
i ztD { R ⋅(j k −l/.. )
= i | R T S
=3
=
i ztD { R ⋅l/.. −! o 2 # ) # ⋅i | R T S
l/..
=
p Q T q ⋅
! rsDtt
! r
sDtt + p ! u C
sDtt
i ] m n
=
i ztD { R + i | R T S
i | R T S (3C )
=
0
 
 
w h er e ! o 2 # ) #  i s t h e cl e ar a n c e r at e of d e a d p ar a sit e s, a n d i d m n  i s t h e c o u nt of m e a s ur e d 
p ar a sit e. If vi a bl e p ar a sit e mi a i s a v ail a bl e t h e n it i s p o s si bilit y t o diff er e nti at e b et w e e n t h e 
killi n g  r at e  a n d  t h e  cl e ar a n c e  r at e.  Ot h er wi s e,  b ot h  p o p ul ati o n  ar e  m e a s ur e d 
i n di sti n g ui s h a bl y, a nd p Q T q  i s fi x e d t o t h e i n vitr o p Q T q  v al u e f or t hi s m o d el. 
  
D uri n g b a s e m o d el b uil di n g, t h e Hill c o effi ci e nt m a y n ot b e i d e ntifi a bl e. T h e n t h e b e st v al u e 
f or t h e Hill c o effi ci e nt will b e d et er mi n e d b y e sti m ati n g m o d el s wit h fi x e d Hill c o effi ci e nt s a n d 
s el e cti n g t h e b e st m o d el b a s e d o n t h e B a y e si a n i nf or m ati o n crit eri o n ( BI C). C o n si d er e d Hill 
c o effi ci e nt s ar e 1, 2, 3, 5, 7, a n d 1 0. If c o v ari at e m o d eli n g i s p erf or m e d, t h e Hill c o effi ci e nt i s 
fi x e d i n t h e v al u e d et er mi n e i n t h e b a s e m o d el b uil di n g. 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 7  of 4 9  
 
IIV will b e a s s u m e d o n all p ar a m et er s b ut t h e Hill c o effi ci e nt. P ar a m et er s ar e a s s u m e d t o b e 
l o g- n or m all y di stri b ut e d b e si d e s t h e l o g-tr a n sf or m e d d e vi ati o n fr o m t h e b a s eli n e p ar a sit e mi a 
v al u e i o & ) )  f or w hi c h n or m al di stri b uti o n will b e a s s u m e d. T h e st a n d ar d d e vi ati o n of t h e II V 
will b e fi x e d t o 0. 2 f or t h e S y s Fit a p pr o a c h. W h e n p ar a m et er s ar e e sti m at e d wit h M o n oli x, 
st a n d ar d d e vi ati o n s of t h e II V will b e e sti m at e d. 
U si n g t h e S y s Fit a p pr o a c h f or P D m o d elli n g, p ar a m et er e sti m ati o n will b e st art e d fr o m 2 4 
i niti al g u e s s e s. I d e ntifi a bilit y of t h e b e st fit will b e a n al y s e d u si n g t h e pr ofil e li k eli h o o d m et h o d. 
1 0. 7.    
D eri v ati o n of s e c o n d ar y effi c a c y p ar a m et er s  
T h e mi ni m u m i n hi bit or y c o n c e ntr ati o n ( MI C), t h e mi ni m al p ar a siti ci d al c o n c e ntr ati o n 
( M P C 9 0), a n d t h e P RR 4 8 ar e d eri v e d fr o m t h e P D m o d el s. T h e MI C i s d efi n e d a s t h e 
c o n c e ntr ati o n w h e n p ar a sit e cl e ar a n c e b y t h e dr u g e q u al s t h e p ar a sit e gr o wt h, i. e., t h e ti m e 
at w hi c h t h e mi ni m u m p ar a sit e c o n c e ntr ati o n i s o b s er v e d. It i s c al c ul at e d wit h t h e f oll o wi n g 
e q u ati o n  i n c a s e of t h e E m a x m o d el. 
}; !
= p ! u C ~
j k
p Q T q + j k 
e / sDtt
 
F or t h e ot h er m o d el s, t h e c al c ul ati o n i s o nl y v ali d at st e a d y- st at e. H o w e v er, a n a p p ar e nt MI C 
i s d et er mi n e d b y si m ul ati o n s a s t h e c o n c e ntr ati o n ! r  at t h e ti m e w h e n t h e pr e di ct e d p ar a site 
c o u nt s ar e at t h e mi ni m u m. 
T h e M P C 9 0 i s d efi n e d a s t h e c o n c e ntr ati o n at w hi c h t h e cl e ar a n c e eff e ct i s at 9 0 % of t h e 
m a xi m u m. It i s c al c ul at e d a s f oll o w s f or t h e E m a x m o d el. 
} i ! Å C = p ! u C ⋅9 e / sDtt  
F or t h e ot h er m o d el s, t h e c al c ul ati o n i s o nl y v ali d at st e a d y- st at e. H o w e v er, a n a p p ar e nt 
M P C 9 0 i s d et er mi n e d b y si m ul ati o n s a s t h e bl o o d c o n c e ntr ati o n ! r  at t h e ti m e w h e n t h e 
pr e di ct e d cl e ar a n c e eff e ct i s at 9 0 % of t h e m a xi m u m d uri n g t h e dr u g eli mi n ati o n p h a s e. 
T h e P R R 4 8 i s d efi n e d a s t h e p ar a sit e cl e ar a n c e a c hi e v e d wit hi n 4 8 h o ur s, u s u all y gi v e n a s 
t h e r e d u cti o n of v al u e s o n l o g 1 0 tr a n sf or m e d s c al e. T h e m a xi m u m c a p a cit y of p ar a sit e 
cl e ar a n c e i n 4 8 h o ur s i s d et er mi n e d a s f oll o w s a s s u mi n g t h at c o n c e ntr ati o n s ar e m ai nt ai n e d 
w ell a b o v e t h e M P C 9 0 f or t hi s ti m e s p a n.  
i k k É Ñ = 4 8 ℎp Q T q −j k
l n(1 0 )
 
T h e a ct u al P R R 4 8 f or a gi v e n d o s e i s d et er mi n e d a s t h e diff er e n c e of pr e di ct e d p ar a sit e 
l e v el s b et w e e n t h e ti m e w h e n t h e d o s e i s gi v e n a n d 4 8 h o ur s l at er. 
K e y effi c a c y p ar a m et er s will b e d eri v e d u si n g t h e si m ul ati o n s of vi a bl e p ar a sit e s i n c a s e t h e 
cl e ar a n c e m o d el i s s el e ct e d a s fi n al m o d el. 
1 0. 8.    
P o st -h o c A n al y si s  
Si m ul ati o n s of cli ni c al effi c a c y p ar a m et er s a n d e n d p oi nt s i n Afri c a n c hil dr e n  
Si m ul ati o n s will b e p erf or m e d u si n g t h e alr e a d y a v ail a bl e  P K m o d el e st a bli s he d a n d p ati e nt s  
Si m ul ati o n s will b e p erf or m e d f or 1, 2 a n d 3 d ail y d o s e r e gi m e n s at t h e a d ult e q ui v al e nt 
7 2 0 m g P yr o n ari di n e t etr a p h o s p h at e. F or e a c h d o s e l e v el, 2 0 0 tri al s wit h 6 2 5 p arti ci p a nt s will 
b e p erf or m e d. D o s e l e v el s, n u m b er of tri al s a n d n u m b er of p arti ci p a nt s m a y b e a dj u st e d if 
a d vi s a bl e. F or e a c h tri al a n e w s et of p o p ul ati o n p ar a m et er s will b e s a m pl e d fr o m t h e 
u n c ert ai nt y di stri b uti o n a n d i n di vi d u al p ar a m et er s w er e s a m pl e d fr o m t h e II V. T h e c o v ari at e s 
a g e, b o d y w ei g ht a n d b a s eli n e p ar a sit e mi a will b e s a m pl e d fr o m a cli ni c al d at a s et of r e al 
p ati e nt s.  
T h e b a s eli n e p ar a sit e mi a i n p ati e nt s ar e t y pi c al 1 0 0 0 f ol d hi g h er t h a n i n c h all e n g e 
p arti ci p a nt s a n d i n t h e r a n g e of 1 0 4  p/ u L  
T h e f oll o wi n g p ar a m et er s will b e c al c ul at e d ( m e di a n a n d 9 5 % CI): 
•  
C m a x: m a xi m u m P yr o n ari di n e bl o o d c o n c e ntr ati o n 
•  
A U C: Ar e a u n d er t h e P yr o n ari di n e bl o o d c o n c e ntr ati o n c ur v e 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 8  of 4 9  
 
•  
P R R 4 8: P ar a sit e r e d u cti o n r ati o aft er 4 8 h 
•  
P ar a sit e cl e ar a n c e ti m e: ti m e t o r e a c h t h e mi cr o s c o pi c L O D ( 1 0 p/ u L) 
•  
E T F C S off: E arl y tr e at m e nt f ail ur e cli ni c al s y m pt o m s off; P ar a sit e mi a o n d a y 2 hi g h er t h a n 
o n d a y 0, irr e s p e cti v e of a xill ar y t e m p er at ur e, or, p ar a sit e mi a o n d a y 3, or, p ar a sit e mi a o n 
d a y 3 > = 2 5 % of c o u n d o n d a y 0. 
•  
E T F C S o n: E arl y tr e at m e nt f ail ur e cli ni c al s y m pt o m s o n; P ar a sit e mi a o n d a y 2 hi g h er t h a n 
o n d a y 0, irr e s p e cti v e of a xill ar y t e m p er at ur e, or, p ar a sit e mi a o n d a y 3 > = 2 5 % of c o u n d 
o n d a y 0.  
•  
L C F 2 8: L at e cli ni c al f ail ur e at d a y 2 8: P ar a sit e mi a a b o v e L L O Q o n a n y d a y b et w e e n d a y 
4 a n d d a y 2 8 i n p ati e nt s w h o di d n ot pr e vi o u sl y m e et a n y of t h e cr it eri a of E T C C S o n. 
•  
L P F 2 8  C S off:  L at e  p ar a sit ol o gi c al  f ail ur e  at  d a y  2 8  cli ni c al  s y m pt o m s  off:  P ar a sit e mi a 
a b o v e L L O Q o n a n y d a y b et w e e n d a y 7 a n d d a y 2 8 wit h a xill ar y t e m p er at ur e < 3 7. 5° C i n 
p ati e nt s w h o di d n ot pr e vi o u sl y m e et a n y of t h e crit eri a of E T F C S off.  
•  
L P F 2 8  C S o n:  L at e  p ar a sit ol o gi c al  f ail ur e  at  d a y  2 8  cli ni c al  s y m pt o m s  o n:  P ar a sit e mi a 
a b o v e L L O Q o n a n y d a y b et w e e n d a y 7 a n d d a y 2 8 wit h a xill ar y t e m p er at ur e < 3 7. 5° C i n 
p ati e nt s w h o di d n ot pr e vi o u sl y m e et a n y of t h e crit eri a of E T F C S o n or L C F 2 8. 
•  
A P R 2 8  C S off:  A d e q u at e  p ar a sit ol o gi c al  r e s p o n s e  at  d a y  2 8  cli ni c al  s y m pt o m s  off; 
A b s e n c e of p ar a sit e mi a o n d a y 2 8 irr e s p e cti v e of a xill ar y t e m p er at ur e, i n p ati e nt s w h o di d 
n ot pr e vi o u sl y m e et a n y of t h e crit eri a of E T F C S off or L P F 2 8 C S off. 
• 
A P R 2 8  C S o n:  A d e q u at e  p ar a sit ol o gi c al  r e s p o n s e  at  d a y  2 8  cli ni c al  s y m pt o m s  o n; 
A b s e n c e of p ar a sit e mi a o n d a y 2 8 irr e s p e cti v e of a xill ar y t e m p er at ur e, i n p ati e nt s w h o di d 
n ot pr e vi o u sl y m e et a n y of t h e crit eri a of E T F C S o n or L C F 2 8 or L P F 2 8 C S o n. 
1 0. 9.    
Pr e s e nt ati o n of R e s u lt s 
A st ati sti c al a n d gr a p hi c al a n al y si s of t h e d at a ( b a s e d o n t h e g e n er al a n al y si s d at a s et will 
i n cl u d e t a bl e s a n d fi g ur e s li st e d i n A p p e n di x, T a bl e A. 
 
T h e a n al y si s r e s ult s will b e r e p ort e d i n t h e m ai n b o d y of t h e C S R a n d t e c h ni c al d et ail s will b e 
r e p orte d a s a n a p p e n di x (r ef err e d t o h er e a s t h e ‘t e c h ni c al a p p e n di x’). T h e t e c h ni c al 
a p p e n di x will i n cl u d e t h e cli ni c al d at a u s e d i n t h e a n al y si s, d at a a s s e m bl y, m o d elli n g 
m et h o d s, all r e s ult s, di s c u s si o n s, a n d c o n cl u si o n s. 
 
K e y r e s ult s will i n cl u d e d e s cri pti o n a n d i nt er pr et ati o n of t h e s el e ct e d m o d el, p ar a m et er 
e sti m at e s, a n d di a g n o sti c pl ot s of g o o d n e s s- of -fit a n d si m ul ati o n s a n d will b e i n cl u d e d i n t h e 
m ai n t e xt. Li mit ati o n s of t h e m o d el pr e di cti o n will b e di s c u s s e d i n t h e Di s c u s si o n s e cti o n of 
t h e t e c h ni c al a p p e n di x a n d m e nti o n e d i n t h e m ai n t e xt of t h e C S R. 
 
1 1.  S E C O N D A R Y E N D P OI N T S A N A L Y SI S  
1 1. 1  
P D A n al y si s  
A n al y si s S et: P D A n al y si s S et 
1 1. 1 . 1   
D at a H a n dli n g  
H a n dli n g of r e pli c at e s 
T h e d at a will b e r e c or d e d a s tri pli c at e p ar a sit a e mi a a n d d u pli c at e g a m et o c yt a e mi a v al u e s f or 
e a c h s u bj e ct at e a c h ti m e p oi nt. T h e r e pli c at e d at a will b e s u m m ari s e d b y c al c ul ati n g t h e 
g e o m etri c m e a n of t h e p ar a sit a e mi a a n d p ar a sit e lif e- st a g e d at a (i. e. g a m et o c yt e mi a a n d 
ri n g- st a g e p ar a sit e s) v al u e s p er s u bj e ct a n d ti m e p oi nt, a n d will b e l o g 1 0 tr a n sf or m e d f or 
st ati sti c al a n al y s e s.    


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 2 9  of 4 9  
 
F or a n y r e pli c at e s t h at w er e n o n- d et e ct s ( N D), t h e v al u e will b e s u b stit ut e d wit h 1 p ar a sit e/ m L 
pri or t o c al c ul ati n g t h e g e o m etri c m e a n f or p ar a sit e mi a q u a ntifi e d fr o m q P C R 1 8 s, or a s t h e 
L O D/ 2 of t h e a s s a y f or p ar a sit e lif e- st a g e d at a (i. e. g a m et o c yt e mi a a n d ri n g- st a g e p ar a sit e s). 
1 1. 1. 2    
P ar a sit e cl e ar a n c e ki n eti c s  
 
T h e P ar a sit e R e d u cti o n R ati o ( P R R) a n d c orr e s p o n di n g p ar a sit e cl e ar a n c e h alf- lif e ( P Ct1/ 2 ) of 
a s e x u al p ar a sit e s i s d eri v e d fr o m t h e cl e ar a n c e r at e of p ar a sit a e mi a aft er a d mi ni str ati o n of 
p yr o n ari di n e tr e at m e nt. T h e a n al y si s of P D r e s p o n s e t o i n v e sti g ati o n al a nti m al ari al t h er a p y 
c o n si st s of: 
•   c al c ul ati n g t h e o pti m al p ar a sit e cl e ar a n c e r at e ( sl o p e c o effi ci e nt fr o m t h e l o g-li n e ar 
d e c a y r e gr e s si o n of q P C R d at a) f or e a c h i n di vi d u al, t h e n,  
•   e sti m ati n g d o s e s p e cifi c p ar a sit e cl e ar a n c e r at e a n d 9 5 % c o nfi d e n c e i nt er v al ( CI) b y 
c al c ul ati n g t h e w ei g ht e d a v er a g e sl o p e e sti m at e a n d c orr e s p o n di n g st a n d ar d err or 
( S E) u si n g a n i n v er s e- v ari a n c e m et h o d. 
 
T h e d at a u s e d f or all m o d el s el e cti o n a n d fitti n g c al c ul ati o n s i s c o m pri s e d of t h e l o g 1 0 
tr a n sf or m e d g e o m etri c m e a n of p ar a sit a e mi a p er ti m e p oi nt p er p arti ci p a nt u p t o t h e fir st 
ti m e p oi nt all p ar a sit a e mi a r e pli c at e s ar e N D. If d uri n g i niti al p ar a sit e cl e ar a n c e t h er e ar e n o 
s a m pl e s N D f or a p arti ci p a nt, t h e l a st ti m e p oi nt i n cl u d e d i n t h e a n al y s e s will b e t hr e e 
ti m e p oi nt s aft er t h e mi ni m u m o b s er v e d p ar a sit e mi a v al u e. All s u b s e q u e nt ti m e p oi nt s ar e s et 
t o ‘ mi s si n g’ r e g ar dl e s s of w h et h er p ar a sit a e mi a v al u e s i n cr e a s e d aft er w ar d s d u e t o p ot e nti al 
r e cr u d e s c e n c e. S e n siti vit y a n al y s e s m a y b e p erf or m e d u si n g ot h er t hr e s h ol d s t o d et er mi n e 
p ar a sit e cl e ar a n c e, f or e x a m pl e, b ut n ot li mit e d t o, t h e l o w er li mit of q u a ntifi c ati o n ( L O Q) (i. e. 
t h e l o w er li mit of t h e r e p ort a bl e r a n g e) of 3 2 p ar a sit e s/ m L or li mit of d et e cti o n of 1 1 1 
p ar a sit e s/ m L [ 1], t o a s s e s s r o b u st n e s s of P R R e sti m at e s. 
 
T h e P R R p er 4 8 h o ur s, P R R 4 8 , f or a s e x u al p ar a sit e will b e e sti m at e d u si n g t h e sl o p e of t h e 
o pti m al fit of t h e l o g -li n e ar r el ati o n s hi p of t h e p ar a sit a e mi a d e c a y o v er ti m e fr o m p yr o n ari di n e 
a d mi ni str ati o n f or e a c h i n di vi d u al, a s d et ail e d i n C T M QI M R S O P 4 1 a n d M ar q u art et al. [ 8].  
 
 
T a bl e  1: S y m b ol s a n d d efi niti o n s of t er m s u s e d i n p ar a sit e cl e ar a n c e a n al y si s ( P art 1) 
 
S y m b ol 
D efi niti o n  
P R R 4 8,i  
I n di vi d u al s p e cifi c P ar a sit e R e d u cti o n R ati o p er 4 8 h 
P R R 4 8, D  
D o s e s p e cifi c P ar a sit e R e d u cti o n R ati o p er 4 8 h 
W e ,D  
Sl o p e c o effi ci e nt of l o g 1 0 p ar a sit a e mi a v s. ti m e pr ofil e f or i n di vi d u al / 
A p V á ,_  
S E c orr e s p o n di n g t o W e ,D f or i n di vi d u al / 
W ̅e ,|   
D o s e s p e cifi c a v er a g e sl o p e e sti m at e 
A p Z W ̅e ,| `  
W ei g ht e d S E of d o s e s p e cifi c a v er a g e sl o p e e sti m at e 
P Ct ½  
P ar a sit e cl e ar a n c e h alf- lif e 
 
R e gr e s si o n M o d elli n g t o D et er mi n e O pti m al Fit  
A r e gr e s si o n m o d elli n g a p pr o a c h t o r e m o v e p ot e nti al l a g a n d/ or t ail p h a s e s of t h e 
p ar a sit a e mi a d e c a y pr ofil e will b e u s e d t o d et er mi n e t h e o pti m al l o g-li n e ar d e c a y r e gr e s si o n. 
T h e al g orit h m c o n si d er s r e m o vi n g p ar a sit a e mi a d at a p oi nt s i n a n it er ati v e pr o c e s s fr o m b ot h 
e n d s of t h e p ar a sit a e mi a c ur v e, i. e. a c o m bi n ati o n of ri g ht c e n s ori n g (r e m o vi n g v al u e s fr o m 
t h e t ail p h a s e) a n d l eft c e n s ori n g (r e m o vi n g v al u e s fr o m t h e l a g p h a s e), a n d u s e s m o d el 
s el e cti o n t e c h ni q u e s t o fi n d t h e o pti m al l o g-li n e ar r e gr e s si o n.  
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 0  of 4 9  
 
T h e al g orit h m t o o bt ai n t h e l o g -li n e ar d e c a y f or e a c h p arti ci p a nt i s b a s e d o n t h e l o g-li n e ar 
r e gr e s si o n d et ail e d i n E q u ati o n ( 1), w h er e Ti m e i s t h e n u m b er of h o ur s si n c e a d mi ni str ati o n 
of a nti m al ari al tr e at m e nt (â/ + & = 1 ,… ,+ ), a n d W C  a n d W e  ar e t h e i nt er c e pt a n d sl o p e 
e sti m at e s, r e s p e cti v el y.   
l o ge C i # ) # -/3 & +/ # = W C + W e â/ + &    ( 1) 
B a s e d o n t h e p ar a sit a e mi a d at a f or e a c h p arti ci p a nt, t h e it er ati v e al g orit h m t o d et er mi n e t h e 
o pti m al l o g -li n e ar d e c a y f or e a c h p arti ci p a nt i s s u m m ari s e d i n T a bl e 2. T h e it er ati v e al g orit h m 
i s c o nti n u e d u ntil a mi ni m u m of f o ur o b s er v ati o n s ar e a v ail a bl e.  
T h e o pti m al l o g -li n e ar r e gr e s si o n m o d el f or a p arti ci p a nt i s d e e m e d a n a p pr o pri at e fit if t h e 
o v er all m o d el p- v al u e < 0. 0 0 1.  
 
  
 
T a bl e 1:  It er ati o n pr o c e s s t o d et er mi n e t h e o pti m al l o g-li n e ar d e c a y c ur v e. 
 
St e p 1: F or e a c h p arti ci p a nt , fit t h e f ull m o d el - fit a li n e ar r e gr e s si o n ( a s d efi n e d b y 
E q u ati o n 1) t o all +  p ar a sit a e mi a  v al u e s of p arti ci p a nt /  
St e p 2: Fit t w o m o d el s: 
( a)  
Fit li n e ar r e gr e s si o n m o d el t o + −1  p ar a sit a e mi a  v al u e s, r e m o vi n g t h e fir st 
o b s er v ati o n. 
( b)  
Fit li n e ar r e gr e s si o n m o d el t o + −1  p ar a sit a e mi a  v al u e s, r e m o vi n g t h e l a st 
o b s er v ati o n. 
St e p 3: D et er mi n e t h e b e st m o d el of St e p 2( a) a n d St e p 2( b), d efi n e d a s t h e m o d el 
c orr e s p o n di n g t o t h e mi ni m u m o v er all m o d el p- v al u e.  
St e p 4: Of t h e b e st m o d el d efi n e d i n St e p 3, r e p e at St e p 2 a n d St e p 3 i n a n it er ati v e 
pr o c e s s u ntil a mi ni m u m of f o ur o b s er v ati o n s. 
St e p 5: Of t h e + −3  b e st m o d el s s el e ct e d p er it er ati o n (i n cl u di n g t h e f ull m o d el ( St e p 
1)), t h e o pti m al m o d el w a s d efi n e d b y t h e mi ni m u m o v er all m o d el p- v al u e.  
 
 
E sti m a ti n g P arti ci p a nt S p e cifi c P R R  
 
T h e sl o p e a n d c orr e s p o n di n g S E e sti m at e of t h e o pti m al li n e ar r e gr e s si o n m o d el i s u s e d t o 
c al c ul at e t h e p arti ci p a nt s p e cifi c P R R 4 8  e sti m at e a n d c orr e s p o n di n g 9 5 % c o nfi d e n c e i nt er v al 
( 9 5 % CI), a s s h o w n i n E q u ati o n ( 2) a n d ( 3), r e s p e cti v el y.  
 
i k k É Ñ ,D = 1 0 é É Ñ × V á ,_                   ( 2) 
9 5 % CI: 1 0 é É Ñ ê V á ,_± e .Å ì × î ï Z V á ,_` ñ   ( 3) 
 
w h er e W e ,D a n d A p V (e ,D) ar e t h e sl o p e a n d c orr e s p o n di n g st a n d ar d err or of t h e sl o p e p ar a m et er 
of t h e o pti m al li n e ar r e gr e s si o n m o d el, r e s p e cti v el y. 
 
E sti m ati n g D o s e S p e cifi c P R R ( P R R 4 8, D ) 
 
Of t h e s p arti ci p a nt s wit h a p pr o pri at e o v er all fit ( p < 0. 0 0 1), t h e a v er a g e P R R
4 8  a n d 
c orr e s p o n di n g 9 5 % CI f or e a c h c o h ort i s e sti m at e d b y u si n g t h e i n v er s e v ari a n c e m et h o d t o 
c al c ul at e t h e w ei g ht e d a v er a g e li n e ar r e gr e s si o n sl o p e (W eóóó) a n d c orr e s p o n di n g S E. T h e 
w ei g ht e d a v er a g e sl o p e f or  -  p arti ci p a nt s i n t h e d o s e wit h a p pr o pri at e o v er all fit i s gi v e n b y 
E q u ati o n ( 4):  
W eóóó =
∑
Z ô _× V á ,_`
ö
_õ á
∑
ô _
ú
_õ á  
, / = 1 ,… ,-      ( 4) 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 1  of 4 9  
 
w h er e t h e w ei g ht i s t h e i n v er s e of t h e s q u ar e d st a n d ar d err or, ù D =
e
î ï Z V á ,_` g  . T h e st a n d ar d 
err or of W eóóó i s e sti m at e d a s, A p Z W eóóó` = û
e
∑
ô _
ö
_õ á  
 .  
 
T h er ef or e t h e d o s e s p e cifi c P R R ( P R R 4 8, D ) a n d c orr e s p o n di n g 9 5 % c o nfi d e n c e i nt er v al i s 
e sti m at e d a s s h o w n i n E q u ati o n ( 5) a n d ( 6), r e s p e cti v el y: 
 
 
i k k É Ñ ,| = 1 0 é É Ñ × V á
óóóó                         ( 5) 
9 5 %  !; :  1 0 é É Ñ × ê V á
óóóó  ± e .Å ì × î ï Z V á
óóóó ` ñ       ( 6) 
 
P ar a sit e Cl e ar a n c e H alf Lif e 
T h e p ar a sit e cl e ar a n c e h alf –lif e ( P Ct ½) will b e d eri v e d fr o m t h e o pti m al d e c a y r at e. T h e 
r el ati o n s hi p b et w e e n P R R4 8  a n d p ar a sit e cl e ar a n c e h alf-lif e (P C 3½) i s a si m pl e tr a n sf or m ati o n 
of t h e P R R 4 8  a s s h o w n i n E q u ati o n ( 7):  
 
i !3 ½ = . * %e C (2 ) × ~
É Ñ  § d • c n
td ¶ á ß Z ® © © ™´ ,_` =
¨ ≠Æá ß (K )
é V á ,_        ( 7) 
 
w h er e P R R 4 8,i  i s t h e p ar a sit a e mi a  r ati o e sti m at e d o v er a 4 8- h o ur i nt er v al t h at i s s u b s e q u e ntl y 
tr a n sf or m e d i nt o a p er h o ur cl e ar a n c e r at e.  
 
C o m p ari s o n of D o s e S p e cifi c P R R 4 8  
T o d et er mi n e w h et h er t h er e ar e diff er e n c e s b et w e e n d o s e s p e cifi c P R R s, a n o m ni b u s t e st f or 
b et w e e n gr o u p diff er e n c e s i s u s e d. T h e t e st i s u s e d t o a s s e s s w h et h er t h er e ar e diff er e n c e s 
i n t h e w ei g ht e d m e a n sl o p e of t h e Ø d o s e s, u si n g t h e t e st st ati sti c s h o wn i n E q u ati o n ( 8),  
 
∞± = ∑
ù Y .Z W ̅Y .−W ̅..`
K
≤
Y ≥e
~ µ ≤é e
K
                 a = 1 ,… ,Ø       ( 8) 
 
T h e w ei g ht f or t h e  a∂§  d o s e i s d e n ot e d b y ù Y .= ∑
ù D Y
n [
D≥e
 f or p arti ci p a nt / wit h a p pr o pri at e 
o v er all fit i n d o s e a. T h e W ̅Y . i s t h e w ei g ht e d a v er a g e sl o p e f or d o s e a a s d efi n e d i n E q u ati o n 
( 9), a n d W ̅.. i s t h e w ei g ht e d gr a n d m e a n gi v e n b y: 
 
W ̅..=
∑
ô [ .V∑[ .  
∏
[ õ á
∑
ô [ .
∏
[ õ á
         ( 9) 
P o st- h o c p air- wi s e c o m p ari s o n s c a n b e c al c ul at e d u si n g t h e t e st st ati sti c π ∫=
∫
ª { º  , w h er e j  
i s t h e c o ntr a st (j = < e W ̅e .+ ⋯+ <≤W ̅≤) a n d 4 ∫ i s t h e v ari a n c e of t h e c o ntr a st (4 ∫=
r ág
ô á . +  … +
r ∏
g
ô ∏.). T h e p- v al u e of t h e o  p air - wi s e c o m p ari s o n s c a n b e c al c ul at e d u si n g t h e S c h eff e m et h o d, 
b y c o m p ari n g π ∫
K  t o a c hi- s q u ar e d di stri b uti o n wit h o −1  d e gr e e s of fr e e d o m. 
 
P ar a sit e R e gr o wt h 
 
I n ci d e n c e of p ar a sit e r e gr o wt h will b e r e p ort e d a s t h e n u m b er a n d p er c e nt a g e of p arti ci p a nt s 
i n e a c h d o s e gr o u p, wit h r e gr o wt h d efi n e d a s ≥5 0 0 0 p ar a sit e s/ m L a n d a t w o-f ol d 
p ar a sit a e mi a i n cr e a s e wit hi n 4 8 h o ur s aft er i niti al r e d u cti o n i n p ar a sit a e mi a. 
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 2  of 4 9  
 
1 1. 1. 3    
Pr e s e nt ati o n of R e s ult s  
P ar a sit a e mi a d at a will b e li st e d f or all e nr oll e d s u bj e ct s a n d s u m m ari s e d f or t h e P D 
p o p ul ati o n. P D p ar a m et er s will b e pr e s e nt e d f or all s u bj e ct s i n cl u d e d i n t h e P D p o p ul ati o n.  
 
P ar a sit e lif e- st a g e d at a ( pf s 2 5, pf M G E T a n d s b p 1 q R T- P C R) will b e li st e d b y p arti ci p a nt a n d 
ti m e p oi nt. 
 
A f ull r e p ort o n t h e p h ar m a c o d y n a mi c d at a a n al y si s will b e i n cl u d e d i n t h e a p p e n di c e s of t h e 
C S R.  T hi s r e p ort will i n cl u d e a li sti n g of P D e n d p oi nt s ( P R R 4 8 , P Ct 1/ 2), a t a bl e f or Su m m ar y of 
P D e n d p oi nt s b y d o s e gr o u p a n d a fi g ur e f or i n di vi d u al l o g- p ar a sit a e mi a r e gr e s si o n fit 
 
Li st of pl a n n e d t a bl e s , li sti n g s a n d fi g ur e s  ar e i n A p p e n di x, T a bl e A. 
 
1 1. 2  
P K A n al y si s  
A n al y si s S et: P K A n al y si s S et 
 
1 1. 2. 1  S o ur c e of d at a  
Bl o o d c o n c e ntr ati o n of P yr o n ari di n e, tr e at m e nt a n d bl o o d s a m pli n g i nf or m ati o n will b e r e c or d e d 
a n d m er g e d t o pr o d u c e P K c o n c e ntr ati o n d at a f or a n al y si s. 
1 1. 2. 2  I m p ut ati o n of N o n-N u m eri c al or N e g ati v e V al u e s  
T h e  i m p ut ati o n  of  n o n - n u m eri c al  or  n e g ati v e  v al u e s  r e p ort e d  i n  t h e  i n p ut  d at a  s et  will  b e 
p erf or m e d a s f oll o w s: 
•  
Pr e - d o s e s a m pl e ti m e s will b e e nt er e d a s z er o 
•   
V al u e s t h at ar e b el o w t h e li mit of q u a ntifi c ati o n ( B L Q) o bt ai n e d pri or t o t h e Cm a x  will b e 
e nt er e d a s z er o 
•   
V al u e s t h at ar e B L Q aft er t h e Cm a x  will b e tr e at e d a s mi s si n g 
•   
P h ar m a c o ki n eti c all y pl a u si bl e c o n c e ntr ati o n v al u e( s) b el o w L L O Q at ti m e p oi nt s 
b et w e e n t w o m e a s ur a bl e c o n c e ntr ati o n v al u e s ar e r e pl a c e d b y t h e L L O Q/ 2 v al u e, 
fl a g g e d a n d i n cl u d e d i n t h e P K e v al u ati o n. I n t h e e v e nt of a n a n o m al o u s b el o w L L O Q 
r e s ult mi d pr ofil e, t h e d at a will b e i nt err o g at e d b y t h e p h ar m a c o ki n eti ci st a n d r ai s e d 
wit h t h e cli e nt b ef or e b ei n g alt er e d. 
•   
S h o ul d p arti al A U C s b e r e q uir e d t h e n v al u e s t h at ar e B L Q aft er Cm a x  m a y b e i m p ut e d 
a s z er o f or t h e s e p arti al ar e a s if l a m b d a- z c a n n ot b e d et er mi n e d 
•   
V al u e s t h at ar e q u a ntifi a bl e aft er at l e a st 2 c o n s e c uti v e B L Q v al u e s aft er Cm a x  will b e 
tr e at e d a s mi s si n g f or t h e c al c ul ati o n of P K p ar a m et er s 
•   
A ct u al ti m e fr o m d o s e a d mi ni str ati o n will b e u s e d f or p ar a m et er e sti m ati o n e x c e pt f or 
t h e pr e-(fir st) d o s e d at a p oi nt w hi c h will b e s et t o t h e n o mi n al ti m e of 0 h o ur. D at a wit h 
mi s si n g ti m e i nf or m ati o n will b e di s c ar d e d. 
•   
V al u e s t h at ar e r e p ort e d a s “ N o R e s ult” or “ N o S a m pl e” et c. will b e tr e at e d a s mi s si n g 
 
1 1. 2. 3  R ul e s f or P h ar m a c o ki n e ti c P ar a m et er E sti m ati o n u si n g Wi n N o nli n 
Bl o o d  c o n c e ntr ati o n  v s  ti m e  pr ofil e s  of  p yr o n ari di n e  will  b e  g e n er at e d  f or  e a c h  s u bj e ct. 
P h ar m a c o ki n eti c  p ar a m et er s  will  b e  e sti m at e d  u si n g  st a n d ar d  P h o e ni x  Wi n N o nli n  m et h o d s, 
d et ail s  of  w hi c h  m a y  b e  f o u n d  i n  t h e  d o c u m e nt ati o n  a c c o m p a n yi n g t h e  Wi n N o nli n  s oft w ar e 
p a c k a g e. T h e f oll o wi n g c o n str ai nt s will a p pl y: 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 3  of 4 9  
 
P ar a m et er E sti m ati o n  
C o n str ai nt  
Tr a p e z oi d al M et h o d  
Li n e ar tr a p e z oi d al li n e ar/l o g i nt er p ol ati o n m et h o d  
N u m b er of P oi nt s u s e d f or L a m b d a- z  
At l e a st 3, n ot i n cl u di n g C m a x  
Mi ni m u m R e q uir e m e nt s f or A U C  
At l e a st 3 c o n s e c uti v e q u a ntifi a bl e c o n c e ntr ati o n s  
D o s e  
A s s u p pli e d  
S a m pli n g Ti m e s  
A ct u al s a m pli n g ti m e s (f or fi n al a n al y si s)  
W h er e p o s si bl e, t h e eli mi n ati o n r at e c o n st a nt (l a m b d a- z) will b e c al c ul at e d f or all s u bj e ct s. T h e 
v al u e  of  l a m b d a- z  will  b e  d et er mi n e d  b y  t h e  sl o p e  of  t h e  r e gr e s si o n  li n e  of  t h e  n at ur al  l o g 
tr a n sf or m e d c o n c e ntr ati o n s v s ti m e.  
T h e c h oi c e of d at a p oi nt s f or d et er mi n ati o n of l a m b d a - z will b e a p pli e d b y t h e P h o e ni x s oft w ar e 
a s a d ef a ult m et h o d, t h e p h ar m a c o ki n eti ci st w h o m a y a dj u st t h e s el e cti o n t o pr o vi d e a m or e 
a p pr o pri at e fit a n d r e c or d s of t hi s will b e d o c u m e nt e d i n t h e s oft w ar e d at a.  
 
1 1. 2. 4  D at a Q u alit y  
T h e f oll o wi n g fl a g s/f o ot n ot e s m a y b e a p pli e d t o t h e p h ar m a c o ki n eti c p ar a m et er s: 
Fl a g  
F o ot n ot e  
a 
R s q of r e gr e s si o n w a s < 0. 8 
b 
P eri o d u s e d f or r e gr e s si o n a n al y si s w a s l e s s t h a n 2-f ol d t h e c al c ul at e d h alf- lif e 
c 
E xtr a p ol at e d p orti o n of A U C 0- i nf > 2 0 %  
d 
I n s uffi ci e nt p o st- Cm a x  d at a p oi nt s f or e sti m ati o n of l a m b d a- z 
e 
E ntir e pr ofil e B L Q, n o p h ar m a c o ki n eti c p ar a m et er s c o ul d b e c al c ul at e d 
f 
R e gr e s si o n c o ul d n ot b e fitt e d 
 
1 1. 2. 5  P K p ar a m et er s  
T h e f oll o wi n g p ar a m et er s will b e e sti m at e d: 
 
C DI S C 
Ter m  
P ara meter  
Defi niti o n 
D P or S F 
N o. of 
D P/ S F  
T M A X  
T ma x  
Ti me of ma xi m u m o bser ve d 
c o nce ntrati o n  
D P  
2 
T L A G  
T la g 
T he dela y bet wee n ti me of d osi n g a n d 
t he a p peara nce of c o nce ntr ati o n  
D P  
2 
C M A X  
C ma x  
Ma xi m u m o bser ve d c o nce ntrati o n  
S F  
3  
A U C L S T  
A U C last 
Area u n der t he c ur ve fr o m 0 ti me t o 
t he last meas ura ble c o nce ntrati o n 
S F  
3 
A U CI F O  
 
A U C 0- i nf 
Area u n der t he c ur ve fr o m 0 ti me 
e xtra p olate d t o i nfi nit y 
S F  
3 
L A M Z H L  
t1/ 2 
A p pare nt eli mi nati o n half -life 
D P  
2  
L A M Z  
La m b da-z 
Sl o pe of t he a p pare nt eli mi nati o n 
p hase  
D P  
4 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 4  of 4 9  
 
C DI S C 
Ter m  
P ara meter  
Defi niti o n 
D P or S F 
N o. of 
D P/ S F  
C L F O  
C L/ F  
T otal b o d y cleara nce after 
e xtra vasc ular a d mi nistrati o n  
S F  
3  
V Z F O  
V z/ F 
A p pare nt v ol u me of distri b uti o n 
base d o n t he ter mi nal p hase after 
e xtra vasc ular a d mi nistrati o n 
S F  
3  
D P = deci mal places 
S F =si g nifica nt fi g ures  
 
1 1. 2. 6  Pr e s e nt ati o n of R e s ult s  
 
All  bi o a n al yti c al  d at a  will  b e  li st e d  a n d  s u m m ari s e d  a c c or di n g  t o  t h e n o mi n al  s a m pli n g 
ti m e p oi nt.  
 
All  p h ar m a c o ki n eti c  p ar a m et er s  g e n er at e d  will  b e  li st e d  a n d  s u m m ari s e d. 
W h er e 
p h ar m a c o ki n eti c  d at a  f ail s  t o  m e et  t h e  d efi n e d  crit eri a  i n  S e cti o n  1 1. 2. 4.  ( D at a  Q u alit y)  t h e 
aff e ct e d r e s ult s will b e e x cl u d e d fr o m t h e d e s cri pti v e st ati sti c s will b e fl a g g e d a s p er e x cl u si o n 
r e a s o n. 
Bi o a n al yti c al a n d P K P ar a m et er s Li sti n g s  
All  bi o a n al yti c al  a n d  p h ar m a c o ki n eti c  N C A  p ar a m et er s  g e n er at e d  will  b e  li st e d  i n  i n di vi d u al 
t a bl e s a c c or di n g t o t h e a n al yt e a n d tr e at m e nt. T h e d at a li sti n g s will b e g e n er at e d a s d efi n e d 
b y t h e P K C o n c e ntr ati o n p o p ul ati o n. 
Bi o a n al yti c al a n d P h ar m a c o ki n eti c S u m m ar y T a bl e s  
Bi o a n al yti c al a n d P K p ar a m et er s s u m m ar y t a bl e s will b e p erf or m e d b y S S R. 
 
S u m m ar y st ati sti c s (i. e., m e a n, m e di a n, S D, C V %, mi ni m u m, m a xi m u m, n, g e o m etri c m e a n, 
g e o m etri c  S D  a n d  g e o m etri c  C V %)  will  b e  c al c ul at e d  f or  P K  r e s ult s  f or  e a c h  ti m e  p oi nt, 
p ar a m et er a n d tr e at m e nt.  
 
All  s u m m ar y  st ati sti c s  (i. e.,  m e a n,  m e di a n,  S D,  C V %,  mi ni m u m,  m a xi m u m  a n d  n)  will  b e 
pr e s e nt e d f or all P K p ar a m et er s f or bl o o d  b y tr e at m e nt. Al s o, g e o m etri c m e a n, g e o m etri c S D 
a n d g e o m etri c C V % will b e pr e s e nt e d f or all P K p ar a m et er s ( e x c e pt T m a x ) b y tr e at m e nt. T h e 
T m a x  s u m m ar y st ati sti c s will b e pr o vi d e d a s n, mi ni m u m, m e di a n, a n d m a xi m u m o nl y. 
 
All t h e d at a s u m m ar y t a bl e s will b e g e n er at e d a s d efi n e d b y t h e P K A n al y si s S et. A d diti o n al 
s u m m ar y t a bl e s mi g ht b e pr o d u c e d if r e q uir e d a n d r e q u e st e d. 
Bi o a n al yti c al a n d P h ar m a c o ki n eti c Fi g ur e s  
All arit h m eti c al m e a n bl o o d  c o n c e ntr ati o n v s. ti m e c ur v e s will b e pr o d u c e d b y tr e at m e nt o n b ot h 
li n e ar/li n e ar a n d l o g1 0 /li n e ar s c al e s. 
 
All  s p a g h etti  pl ot s  of  i n di vi d u al bl o o d   c o n c e ntr ati o n s  a g ai n st n o mi n al   s a m pli n g  ti m e s  aft er 
d o si n g f or e a c h tr e at m e nt will b e pr o d u c e d o n b ot h a li n e ar/li n e ar a n d l o g 1 0 /li n e ar s c al e. E a c h 
v ol u nt e er’ s c o n c e ntr ati o n pr ofil e will b e r e pr e s e nt e d o n t h e s e pl ot s wit h a diff er e nt s y m b ol a n d 
a l e g e n d will b e i n cl u d e d o n t h e pl ot s t o d efi n e t h e s y m b ol s u s e d. 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 5  of 4 9  
 
Li st of s u m m ar y  t a ble s, li sti n g s a n d fi g ur e s ar e i n A p p e n di x, T a bl e A.  
 
 
1 1. 3  
S af et y  
A n al y si s S et: S af et y d at a will u s e b ot h F A S a n d I M P S et s  
 
S af et y a n d t ol er a bilit y will b e a s s e s s e d b y cli ni c al r e vi e w of t h e f oll o wi n g p ar a m et er s:  
•   A d v er s e e v e nt s (A E s )  
•   Vit al si g n s  
•  1 2 -l e a d E C G 
•   H a e m at ol o g y, bi o c h e mi str y, uri n al y si s 
•   P h y si c al e x a mi n ati o n 
1 1. 3. 1  A d v er s e E v e nt s  
A E s will b e s e p ar at e d i nt o t w o c at e g ori e s. 
 
•  
I n o c ul u m- e m er g e nt A E s (I E A E s) = A E s b e gi n ni n g a n y ti m e fr o m i n o c ul ati o n wit h t h e 
c h all e n g e a g e nt u ntil t h e e n d of st u d y. 
 
•  
Tr e at m e nt - e m er g e nt A E s ( T E A E s) = A E s b e gi n ni n g a n y ti m e fr o m a d mi ni str ati o n of 
t h e I M P u ntil t h e e n d of st u d y. 
 
1 1. 3. 1. 1  
A E  Li sti n g s 
All A E d at a will b e li st e d f or e a c h p arti ci p a nt a n d will i n cl u d e: 
•  
I E A E s 
•  
T E A E s  
•   I E A E s r el at e d t o m al ari a 
•  
T E A E s r el at e d t o I M P  
•   I E S A E s ( S eri o u s a d v er s e e v e nt s) 
•  
T E S A E s  
•   I E A E s l e a di n g t o di s c o nti n u ati o n or wit h dr a w al 
•   
T E A E s l e a di n g t o di s c o nti n u ati o n or wit h dr a w al 
•   I E A E SI s ( A d v er s e E v e nt s of S p e ci al I nt er e st) 
•  
T E A E SI s  
 
 
P ar a m et er s t o b e i n cl u d e d i n t h e li sti n g s will b e:  
•   E v e nt T er m 
•  D at e s , ti m e s a n d st u d y d a y s of o n s et a n d r e s ol uti o n 
•   S e v erit y ( C T C A E Gr a d e s 1 t o 5) 
•   O ut c o m e ( N ot R e c o v er e d / N ot R e s ol v e d; R e c o v er e d / R e s ol v e d; R e c o v er e d / R e s ol v e d 
wit h S e q u el a e; R e c o v eri n g / R e s ol vi n g; F at al; U n k n o w n) 
•   R el ati o n s hi p of A E t o I M P ( N ot R el at e d, R el at e d) 
•   R el ati o n s hi p of A E t o r e s c u e m e di c ati o n ( N ot R el at e d, R el at e d) 
•   R el ati o n s hi p of A E t o m al ari a c h all e n g e a g e nt ( N ot R el at e d, R el at e d) 
•   R el ati o n s hi p of A E t o st u d y pr o c e d ur e s ( N ot R el at e d, R el at e d) 
•   A cti o n t a k e n wit h I M P ( N o a cti o n t a k e n; R e s c u e m e di c ati o n a d mi ni st er e d i n st e a d of I M P) 
•   Ot h er a cti o n s t a k e n (i n cl u di n g wit h dr a w al fr o m t h e st u d y) 
•   S eri o u s n e s s ( a n d S eri o u s A d v er s e E v e nt ( S A E) crit eri a) 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 6  of 4 9  
 
•   A E SI st at u s ( Y e s/ N o) 
•   R el at e d n e s s t o C O VI D-1 9 ( Y e s/ N o)  
D eri v e d p ar a m et er s: 
•   D ur ati o n i n h o ur s  
•   Ti m e i n d a y s of o n s et r el ati v e t o fir st I M P a d mi ni str ati o n 
 
 
1 1. 3. 1. 2  
A E T a bl e s  
T h e n u m b er of e v e nt s, a s w ell a s t h e n u m b er a n d p er c e n t a g e of p arti ci p a nt s e x p eri e n ci n g a n 
A E , will b e s u m m ari s e d b y, d o s e gr o u p a n d o v er all a s f oll o w s: 
 
O v er all s u m m ar y of I E A E s ( F A s et)  
O v er all s u m m ar y of T E A E s (I M P s et) 
 
S u m m ar y of I E A E s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m ( F A s et) 
S u m m ar y of T E A E s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m (I M P s et) 
 
S u m m ar y of I E A E s b y S y st e m Or g a n Cl a s s, Pr ef err e d T er m a n d M a xi m u m S e v erit y ( F A s et) 
S u m m ar y of T E A E s b y S y st e m Or g a n Cl a s s, Pr ef err e d T er m a n d M a xi m u m S e v erit y (I M P 
s et) 
 
S u m m ar y of M al ari a- R el at e d I E A E s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m ( F A s et) 
S u m m ar y of I M P- R el at e d T E A E s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m (I M P s et) 
 
S u m m ar y of M al ari a- R el at e d I E A E s b y S y st e m Or g a n Cl a s s, Pr ef err e d T er m a n d M a xi m u m 
S e v erit y ( F A s et) 
S u m m ar y of I M P- R el at e d T E A E s b y S y st e m Or g a n Cl a s s, Pr ef err e d T er m a n d M a xi m u m 
S e v erit y (I M P s et) 
 
S u m m ar y of I E S A E s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m ( F A s et) 
S u m m ar y of T E S A E s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m (I M P s et) 
 
O v er all S u m m ar y of I E A E SI s ( F A s et) 
O v er all S u m m ar y of T E A E SI s (I M P s et) 
 
S u m m ar y of I E A E SI s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m ( F A s et) 
S u m m ar y of T E A E SI s b y S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m (I M P s et 
 
  
F or t h e s u m m ar y t a bl e s , p arti ci p a nt s w h o e x p eri e n c e t h e s a m e A E (i n t er m s of t h e M e d D R A 
pr ef err e d t er m) m or e t h a n o n c e will o nl y b e c o u nt e d o n c e. 
A d diti o n al s u m m ar y t a bl e s of A E s r el at e d t o r e s c u e m e di c ati o n s will b e i n cl u d e d if r e q u e st e d 
b y M M V.  
 
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 7  of 4 9  
 
1 1. 3. 2  C o n c o mit a nt M e di c ati o n  
M e di c ati o n s u s e d i n t hi s st u d y will b e c o d e d u si n g W H O Dr u g Gl o b al v er si o n B 3 S e p 2 0 2 1 – 
A d d e d C o nt e xt. 
 
C o n c o mit a nt m e di c ati o n s ar e d efi n e d a s m e di c ati o n s c o nti n u e d or n e wl y r e c ei v e d at or aft er 
a d mi ni str ati o n of t h e m al ari a c h all e n g e a g e nt t hr o u g h t o t h e E n d of St u d y vi sit. 
 
If a m e di c ati o n h a s a mi s si n g or p arti al mi s si n g st art/ e n d d at e or ti m e a n d it c a n n ot b e 
d et er mi n e d w h et h er it w a s t a k e n b ef or e i niti al tr e at m e nt or c o n c o mit a ntl y, it will b e c o n si d er e d 
a s pri or a n d c o n c o mit a nt. 
 
C o n c o mit a nt m e di c ati o n s will b e s u m m ari s e d b y A T C a n d P N. T h e s u m m ar y t a bl e s will s h o w 
t h e n u m b er a n d p er c e nt a g e of p arti ci p a nt s t a ki n g e a c h m e di c ati o n b y A T C a n d P N. 
 
F or t h e s u m m ari e s of  c o n c o mit a nt m e di c ati o n s, p arti ci p a nt s w h o t a k e t h e s a m e m e di c ati o n 
(i n t er m s of t h e A T C a n d P N) m or e t h a n o n c e will o nl y b e c o u nt e d o n c e f or t h at m e di c ati o n. 
 
U s e of r e s c u e m e di c ati o n s will b e a n al y s e d a s p art of t h e tr e at m e nt e x p o s ur e a n al y si s 
( S e cti o n 8).  
 
 
 


Pr ot o c ol N u m b er: M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y  2 0 2 3 
D o c u m e nt st at u s: S A P V er s i o n 1. 1 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7 /S E P / 2 02 0  
C O N FI D E N TI AL  
 
P a g e 3 8  of 4 9  
 
1 1. 3. 3  L a b or at or y  
P ar a m et er s  
H e m at ol o g y
•   
B a s o p hil s ( a b s ol ut e) 
•  
M o n o c yt e s ( a b s ol ut e) 
•   
E o si n o p hil s ( a b s ol ut e) 
•   
N e utr o p hil s ( a b s ol ut e) 
•   
H a e m at o crit 
•   
Pl at el et c o u nt 
•  
H a e m o gl o bi n  
•   
R e d bl o o d c ell c o u nt 
•   L y m p h o c yt e s ( a b s ol ut e) 
•   
W hit e bl o o d c ell c o u nt 
Bi o c h e mi str y 
•  
Al a ni n e A mi n otr a n sf er a s e ( A L T) 
•   
G a m m a Gl ut a m yl Tr a n s p e pti d a s e 
( G G T) 
•  
Al b u mi n  
•   
Gl u c o s e (r a n d o m a n d f a sti n g) 
•   
Al k ali n e p h o s p h at a s e ( A L P) 
•   
Gl u c o s e- 6- p h o s p h at a s e ( G 6 P D) 
•   
A s p art at e A mi n otr a n sf er a s e ( A S T) 
•   L a ct at e d e h y dr o g e n a s e ( L D H) 
•   
Bi c ar b o n at e 
•  
L D L  
•  
H D L  
•   
Bilir u bi n ( dir e ct) 
•   
P h o s p h at e 
•  
Bilir u bi n (t ot al)  
•   
P ot a s si u m 
•   
C al ci u m 
•  
Pr ot ei n T ot al  
•   
C al c ul at e d e G F R ( C K D-E PI)  
•  
S o di u m  
•   
C- R e a cti v e pr ot ei n  
•   
Tri gl y c eri d e 
•  
C hl ori d e  
•   
Tr o p o ni n T/ hi g h s e n siti vit y 
•  
C h ol e st er ol T ot al  
•  
Ur e a  
•  
Cr e ati ni n e  
•   
Uri c A ci d 
•   
Cr e ati n e ki n a s e 
 
 
Pr e g n a n c y T e st a n d F S H  
 
  
β-H u m a n C h ori o ni c G o n a d otr o pi n ( β - H C G) ( w o m e n of c hil d b e ari n g p ot e nti al o nl y) 
  
F olli c ul ar Sti m ul ati n g H or m o n e ( F S H) ( p o st- m e n o p a u s al w o m e n o nl y) 
   
Uri n al y si s  
Di p sti c k T e sti n g  
Q u a ntit ati v e A s s e s s m e nt s  
Uri n e C h e mi str y  
 
R e d bl o o d  c ell s  
 
R e d bl o o d c ell s  
  
Gl u c o s e 
  
Gl u c o s e 
 
W hit e bl o o d c ell s  
 
Pr ot ei n  
 
K et o n e  
 
 
 
W hit e bl o o d c ell s  
 
 
 
Pr ot ei n   
 
 
 
p H   
 
 
 
R e d bl o o d c ell s  
 
 
  
Gl u c o s e 
 
 
 
K et o n e  
 
 
C o a g ul ati o n  
•   I nt er n ati o n al N or m ali z e d R ati o (I N R) •   
A cti v at e d P arti al T hr o m b o pl a sti n 
Ti m e ( A P T T)  


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 3 9  of 4 9  
 
S er ol o g y  
•   
H e p atiti s B s urf a c e a nti g e n ( H B 
s A G) 
•  
A nti - h e p atiti s B c or e a nti b o di e s 
( a nti- H B c A b) 
•   
H e p atiti s C a nti b o di e s 
•  
R e d bl o o d c ell a nti b o di e s  
•  
H u m a n I m m u n o d efi ci e n c y Vir u s 
( HI V) A g/ A b 1/ 2 ( a nti -HI V 1 a n d 
a nti -HI V 2 A b ) 


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 0  of 4 9  
 
Bi o st ati sti c al m et h o d s  
  
Li sti n g s 
All l a b or at or y p ar a m et er s a n d c h a n g e fr o m tr e at m e nt b a s eli n e v al u e s will b e pr e s e nt e d i n d at a li sti n g s. V al u e s o ut si d e t h e l a b or at or y r ef er e n c e 
r a n g e will b e li st e d wit h fl a g s if c o n si d er e d l o w or hi g h f or v al u e s o ut si d e t h e r ef er e n c e r a n g e s ( w h er e a p pli c a bl e) a n d if v al u e s c o n si d er e d t o b e 
cli ni c all y si g nifi c a nt b y t h e i n v e sti g at or. 
 
Pr e g n a n c y t e st d at a will b e li st e d o nl y , f or all w o m e n.  F S H d at a will b e li st e d o nl y, f or all p o st- m e n o p a u s al w o m e n.  
 
C o a g ul ati o n d at a a s w ell a s q u alit ati v e a n d q u a ntit ati v e uri n al y si s d at a will al s o b e li st e d o nl y. 
 
T a bl e s  
H a e m at ol o g y, bi o c h e mi str y a n d c o nti n u o u s di p sti c k uri n al y si s l a b or at or y d at a will b e s u m m ari s e d f or e a c h s c h e d ul e d vi sit, i n cl u di n g o b s er v e d v al u e s, 
a b s ol ut e c h a n g e fr o m tr e at m e nt b a s eli n e   
 
C at e g ori c al di p sti c k uri n al y si s r e s ult s will b e s u m m ari s e d f or e a c h s c h e d ul e d vi sit u si n g fr e q u e n c y t a b ul ati o n s.  
 
L a b or at or y a b n or m aliti e s will b e pr e s e nt e d a s cr o s s- t a b ul ati o n s of t h e a b n or m alit y at e a c h p o st-tr e at m e nt b a s eli n e a n al y si s vi sit v er s u s t h e tr e at m e nt 
b a s eli n e. N u m b er s of p arti ci p a nt s wit h tr e at m e nt- e m er g e nt a b n or m aliti e s will al s o b e s h o w n.  
 
I n a d diti o n, th e li v er f u n cti o n t e st s, n a m el y A S T, A L T, a n d t ot al bilir u bi n, ar e u s e d t o a s s e s s p o s si bl e dr u g-i n d u c e d li v er t o xi cit y . T h e pr o p orti o n of 
p ati e nt s wit h P ot e nti al Cli ni c all y Si g nifi c a nt A b n or m alit y ( P C S A) v al u e s at a n y p o st b a s e  li n e vi sit will s u m m ari z e d. T h e hi g h e st v al u e of P C S A a n d 
ti m e t o t h e hi g h e st P C S A r e a di n g will al s o b e s u m m ari z e d. A n y of t h e f oll o wi n g e v e nt will b e c o n si d er e d a s a P C S A: 
 
•   
A n y A L T or A S T v al u e a b o v e 5 × U L N. 
•  
A n el e v ati o n i n bilir u b i n 2 × U L N. 
•   
A n y A S T or A L T v al u e a b o v e 2 × U L N a n d (t ot al bilir u bi n > 1. 5 × U L N). 
•   
A n y A S T or A L T v al u e a b o v e 2 × U L N wit h t h e a p p e ar a n c e of f ati g u e, n a u s e a, v o miti n g, ri g ht u p p er q u a dr a nt p ai n or t e n d er n e s s, fe v er, r a s h 
a n d/ or e o si n o p hili a ( e o si n o p hil p er c e nt or c o u nt a b o v e t h e U L N).  
•   
C o n stit ut e s a p o s si bl e H y’ s L a w c a s e 
o   H y’ s L a w c a s e i s d efi n e d a s a P arti ci p a nt wit h a n y v al u e of al a ni n e or a s p art at e a mi n otr a n sf er a s e gr e at er t h a n 3 × U L N t o g et h er wit h a n 
i n cr e a s e i n t ot al bilir u bi n t o a v al u e gr e at er t h a n 2 × U L N a n d n ot a s s o ci at e d t o a n al k ali n e p h o s p h at a s e v al u e gr e at er t h a n 2 × U L N ( F D A 
G ui d a n c e o n Dr u g I n d u c e d Li v er I nj ur y: Pr e m ar k eti n g Cli ni c al E v al u ati o n [ 2 0 0 9]). 


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 1  of 4 9  
 
 
1 1. 4  
Vit al Si g n s  
P ar a m et er s  
•   
S y st oli c Bl o o d Pr e s s ur e ( S B P) ( m m H g) 
•   
Di a st oli c Bl o o d Pr e s s ur e ( D B P) ( m m H g) 
•  
H e art R at e ( b e at s/ mi n ut e)  
•  
T e m p er at ur e (° C)  
•   
R e s pir at or y r at e ( b e at s/ mi n) 
Bi o st ati sti c al m et h o d s  
All v it al si g n s d at a  will b e li st e d f or all p arti ci p a nt s.  A n y v al u e s o ut si d e of t h e pr ot o c ol d efi n e d n or m al r a n g e s will b e fl a g g e d, wit h cli ni c al 
si g nifi c a n c e st at u s pr e s e nt e d f or o ut- of -r a n g e r e s ult s. 
 
T a bl e 2 : Vit al Si g n s N or m al R a n g e s 
P ar a m et er  
R a n g e  
S y st oli c bl o o d pr e s s ur e              
9 0 -1 4 0 m m H g  
Di a st oli c bl o o d pr e s s ur e            
5 0 -9 0 m m H g  
H e art r at e                                  
5 0 -1 0 0 b p m  
T e m p er at ur e                              
3 5. 0 -3 7. 5° C  
R e s pir at or y r at e                         
1 0 - 2 5 br e at h s/ mi n 
 
Vit al si g n p ar a m et er s  will b e s u m m ari s e d b y pr e s e nti n g s u m m ar y st ati sti c s f or o b s er v e d v al u e s a n d c h a n g e fr o m tr e at m e nt b a s eli n e v a l u e s f or 
e a c h s c h e d ul e d vi sit. 
1 1. 5  
B o d y M e a s ur e m e nt s  
D efi niti o n of v ari a bl e s  
•   
H ei g ht ( S cr e e ni n g O nl y) 
•  
W ei g ht  


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 2  of 4 9  
 
•   
B o d y m a s s i n d e x ( B MI) 
Bi o st ati sti c al m et h o d s  
B o d y m e a s ur e m e nt d at a will b e li st e d f or all p arti ci p a nt s a n d vi sit s i n cl u di n g h ei g ht, w ei g ht a n d B o d y M a s s I n d e x ( B MI) w hi c h i s a ut o- c al c ul at e d 
wit hi n t h e C R F.  
 
O b s er v e d v al u e s, a s w ell a s c h a n g e s fr o m tr e at m e nt b a s eli n e, will b e s u m m ari s e d d e s cri pti v el y f or w ei g ht a n d B MI b y vi sit.  
P h y si c al E x a mi n ati o n  
P ar a m et er s  
•  
W ei g ht  
•   
S ki n 
•   
H E E N T ( H e a d, E y e s, E ar s, N o s e, T hr o at) •   
N e ur ol o gi c al / R efl e x e s  
•   
H ei g ht ( s cr e e ni n g o nl y) 
•   
H e art / cir c ul ati o n 
•   
C h e st / L u n g s 
•   
N e c k 
•  
A b d o m e n  
 
Bi o st ati sti c al m et h o d s  
P h y si c al e x a mi n ati o n fi n di n g s will b e li st e d f or all p arti ci p a nt s a n d vi sit s 
1 1. 6  
1 2 -l e a d E C G 
P ar a m et er s  
•    
•   
P R i nt er v al ( m s e c) 
•   
Q R S i nt er v al ( m s e c) 
•   
Q T c F i nt er v al ( m s e c) 
•   
O v er all E C G a s s e s s m e nt ( N or m al, A b n or m al, N ot E v al u a bl e) 
•   
E C G a b n or m alit y ( a s a p pr o pri at e) 
•   
Cli ni c al Si g nifi c a n c e 


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 3  of 4 9  
 
Bi o st ati sti c al m et h o d s  
E C G p ar a m et er s will b e li st e d f or all p arti ci p a nt s a n d vi sit s. Tri pli c at e E C G s will b e pr e s e nt e d i n li sti n g s f or i n di vi d u al r e a di n g s a s w ell a s t h e m e a n 
f or e a c h tri pli c at e. T h e tri pli c at e m e a n s will b e u s e d f or t h e s u m m ar y t a bl e. A n y v al u e s o ut si d e of t h e pr ot o c ol d efi n e d n or m al r a n g e s ( T a bl e X X) 
will b e fl a g g e d.   
 
 
T a bl e 3 : E C G N or m al R a n g e s 
P ar a m et er  
R a n g e  
P R i nt er v al              
≤2 1 0 ms ec  
Q R S            
5 0 – 1 2 0 ms ec  
Q Tc F                            
M al es: ≤4 5 0 ms ec  
F e m al es: ≤4 7 0 ms ec  
 
 
O b s er v e d v al u e s, a s w ell a s c h a n g e s fr o m tr e at m e nt b a s eli n e, will b e s u m m ari s e d d e s cri pti v el y f or all E C G p ar a m et er s b y vi sit. A n a d diti o n t a bl e 
will pr e s e nt t h e fr e q u e n ci e s of p arti ci p a nt s w h o f ulfill t h e f oll o wi n g pr ol o n g ati o n ( c h a n g e fr o m b a s eli n e) crit eri a, c o n si d eri n g all s c h e d ul e d a n d n o n-
s c h e d ul e d vi sit d at a: 
•   
Q T c F pr ol o n g ati o n > 3 0 
•   
Q T c F pr ol o n g ati o n > 6 0 
•   
Q T c F > 4 5 0 m s e c f or m al e s 
•   
Q T c F > 4 7 0 m s e c   f or f em al e s  
•    
 
1 1. 7  
Pr e s e nt ati o n of S af et y R e s ult s  
T h e t a bl e s u m m ar y a n d li sti n g s of A E s a n d l a b p ar a m et er s will b e pr e s e nt e d f or t h e C S R, a n d ar e pr o vi d e d i n A p p e n di x, T a bl e B. 
 
1 2.  E X P L O R A T O R Y P A R A M E T E R S  
N ot A p pli c a bl e  


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 4  of 4 9  
 
1 3.  C H A N G E S T O T H E P L A N N E D A N A L Y SI S  
T h er e i s n o c h a n g e t o t h e pl a n n e d a n al y si s b y t h e ti m e w h e n t hi s S A P w a s writt e n. 
1 4.  I N T E RI M A N D FI N A L A NA L Y SI S  
T hi s st u d y h a s n o f or m al i nt eri m a n al y s e s ot h er t h a n r e vi e w of s af et y, t ol er a bilit y, p ar a sit a e mi a a n d P K p ar a m et er s b y t h e S D R T b et w e e n c o h ort s. 
 
1 5.  S O F T W A R E  
S A S ®  V er si o n 9. 4 or hi g h er ( S A S I n s tit ut e, C ar y, N ort h C ar oli n a, U S A) f or S af et y, d e m o gr a p hi c a n d b a s eli n e c h ar a ct eri sti c s, vol u nt e er 
di s p o siti o n, pr ot o c ol d e vi ati o n s, p ri or m e di c ati o n s a n d t r e at m e nt e x p o s ur e a n al y s e s 
All P K/ P D a n al y s e s will b e p erf or m e d i n R 4. 1. 3 a n d I Q Rt o ol p a c k a g e 1. 1 0. 0 [I Q Rt o ol s]. N L M E m o d elli n g will b e p erf or m e d wit h M o n oli x 
2 0 1 9 R 1 [ M L X 1 9] u si n g St o c h a sti c A p pr o xi m ati o n E x p e ct ati o n M a xi mi z ati o n ( S A E M) f or p ar a m et er e sti m ati o n. F or P D m o d elli n g w h er e 
t y pi c all y t h e v ari a n c e of r a n d o m- eff e ct s c a n n ot b e e sti m at e d fr o m t h e d at a, t h e I Q Rt o ol s S y s Fit a p pr o a c h [ K A S 1 9] will b e u s e d t o e sti m at e 
t y pi c al a n d i n di vi d u al P D p ar a m et er s. 
D at a m a ni p ul ati o n a n d d at a a n al y s e s f or all p h ar m a c o d y n a mi c d at a will b e p erf or m e d u si n g R ( hi g h e st v er si o n a v ail a bl e).  
1 6.  R E F E R E N C E S  
1)   W a n g C Y T, B all ar d E L, P a v a Z M ar q u art L, G a y d o n J, M ur p h y S C, W hil e y D, O’ R o ur k e P, M c C art h y J S ( 2 0 2 1) A n al yti c al v ali d ati o n of a r e al -
ti m e h y dr ol y si s pr o b e P C R a s s a y f or q u a ntif yi n g Pl a s m o di u m f al ci p ar u m p ar a sit e s i n e x p eri m e nt all y i nf e ct e d h u m a n a d ult s, M al ari a J o ur n al , 
2 0( 1 8 1).  
2)   M ar q u art  L,  B a k er  M,  O’ R o ur k e  P,  M c C art h y J S ( 2 0 1 5)  E v al u ati n g  t h e p h ar m a c o d y n a mi c  eff e ct  of  a nti m al ari al dr u g s  i n  cli ni c al  tri al s  b y 
q u a ntit ati v e P C R, A nti mi cr o bri al A g e nt s a n d C h e m ot h er a p y, 5 9, 4 2 4 9 – 5 9.  
3)  I Q Rt o ol s htt p s:// w w w.i nti q u a n. c o m/i qr-t o ol s/  
 
4)  K A S 1 9  K a s c h e k D, M a d er W, F e hli n g- K a s c h e k M, R o s e n bl att M, Ti m m er J. D y n a mi c M o d eli n g, P ar a m et er E sti m ati o n, a n d U n c ert ai nt y 
A n al y si s i n R. J. St at. S oft. 2 0 1 9 A pr 3 0; 8 8( 1 0): 1- 3 2.  
 
5)  M E T 1 1  M et h a n e et h or n J, J u n g D, Fl e c k e nt st ei n L. P o p ul ati o n P h ar m a c o ki n eti c s of P yr o n ari di n e f oll o wi n g Or al P yr o n ari di n e/ Art e s u n at e 
Tr e at m e nt i n H e alt h y a n d M al ari a I nf e ct e d P arti ci p a nt s ( V er si o n 1. 1). 2 0 1 1 A pr 8. 
 
6)  M L X 1 9  htt p://li x oft. c o m/  


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 5  of 4 9  
 
 
7)  R A U 0 9  R a u e A, Kr e ut z C, M ai w al d T, B a c h m a n n J, S c hilli n g M, Kli n g m üll er U, Ti m m er J. Str u ct ur al a n d pr a cti c al i d e ntifi a bilit y a n al y si s of 
p arti all y o b s er v e d d y n a mi c al m o d el s b y e x pl oiti n g t h e pr ofil e li k eli h o o d. Bi oi nf or m ati c s. 2 0 0 9 J u n 8; 2 5( 1 5): 1 9 2 3- 9.  
 
8)  M M V _ P yr o n ari di n e _ VI S _ V er si o n 4. 0 _ 2 7 J ul y 2 0 2 2 _ SI G N E D 
 
9)   e C O S _ U ni q u e C R F s _ U ni q u e P a g e s _M M V _ P yr o n ari di n e _ 2 1 _ 0 1 _ V 1. 0 _ 1 1 M ar 2 0 2 2  
 
 
 
1 7.    
1 8.    


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 6  of 4 9  
 
1 9.  A P P E N DI X  
T a bl e A . P D a n d P K Pl a n n e d T a bl e s , Li sti n g s  a n d Fi g ur e s  
 
Titl e  
A n al y si s S et  
 
 
P D Li sti n g s  
 
P h ar m a c o d y n a mi c – P ar a sit e mi a 
P D  
P h ar m a c o d y n a mi c – P ar a sit e Lif e C y cl e St a g e s  
P D  
P K Li sti n g s  
 
P h ar m a c o ki n eti c C o n c e ntr ati o n s  
P K  
P h ar m a c o ki n eti c P ar a m et er s  
P K  
P D  T a bl e s  
 
S u m m ar y of P h ar m a c o d y n a mi c i n q P C R 1 8 s  
P D  
S u m m ar y of P h ar m a c o d y n a mi c i n P ar a sit e Lif e C y cl e St a g e s   
P D  
P K T a bl e s  
 
S u m m ar y of P h ar m a c o ki n eti c C o n c e ntr ati o n  
P K  
S u m m ar y of P h ar m a c o ki n eti c P ar a m et er s  
P K  
P D Fi g ur e s  
 
I n di vi d u al q P C R 1 8 s-ti m e Pr ofil e s 
P D  
I n di vi d u al R T- P C R pf s 2 5-ti m e Pr ofil e s 
P D  
I n di vi d u al R T-P C R pf M G E T -ti m e Pr ofil e s 
P D  
I n di vi d u al R T-P C R S B P - 1-ti m e Pr ofil e s 
 
P K Fi g ur e s  
 
Arit h m eti c M e a n  ( + S D) P h ar m a c o ki n eti c C o n c e ntr ati o n-ti m e Pr ofil e s 
P K  
I n di vi d u al P h ar m a c o ki n eti c C o n c e ntr ati o n-ti m e Pr ofil e s 
P K  
 
 
T a bl e B . B a s eli n e C h ar a ct eri sti c s a n d S af et y A n al y si s T a bl e s a n d Li sti n g s  


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 7  of 4 9  
 
 
Titl e  
A n al y si s 
S et  
 
 
Li sti n g s  
 
P arti ci p a nt Di s p o siti o n 
F A S  
Tr e at m e nt Gr o u p A s si g n m e nt 
F A S  
Pr ot o c ol D e vi ati o n s 
F A S  
A n al y si s P o p ul ati o n A s si g n m e nt 
F A S  
D e m o gr a p hi c s 
F A S  
M e di c al Hi st or y 
F A S  
B e c k D e pr e s si o n I n d e x ( B DI- 2)  
F A S  
Pr e g n a n c y T e st 
F A S  
F olli cil e Sti m ul ati n g H or m o n e ( F S H) 
F A S  
S er ol o g y S cr e e ni n g 
F A S  
Uri n e Dr u g S cr e e ni n g 
F A S  
Al c o h ol Br e at h T e st  
F A S  
Pri or a n d C o n c o mit a nt M e di c ati o n s 
F A S  
M al ari a C h all e n g e A g e nt A d mi ni str ati o n 
F A S  
St u d y Dr u g A d mi ni str ati o n 
F A S  
R e s c u e M e di c ati o n A d mi ni str ati o n 
F A S  
I n o c ul u m- E m er g e nt A d v er s e E v e nt s 
F A S  
Tr e at m e nt - E m er g e nt A d v er s e E v e nt s 
I M P 
M al ari a C h all e n g e A g e nt r el at e d I n o c ul u m- E m er g e nt A d v er s e E v e nt s 
F A S  
I M P r el at e d Tr e at m e nt- E m er g e nt A d v er s e E v e nt s 
I M P 
I n o c ul u m E m er g e nt S eri o u s A d v er s e E v e nt s 
F A S  
Tr e at m e nt E m er g e nt S eri o u s A d v er s e E v e nt s 
I M P 
I n o c ul u m- E m er g e nt A d v er s e E v e nt s L e a di n g t o Di s c o nti n u ati o n 
F A S  
Tr e at m e nt - E m er g e nt A d v er s e E v e nt s L e a di n g t o Di s c o nti n u ati o n 
I M P 


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 8  of 4 9  
 
I n o c ul u m- E m er g e nt A d v er s e E v e nt s of S p e ci al I nt er e st 
F A S  
Tr e at m e nt - E m er g e nt A d v er s e E v e nt s of S p e ci al I nt er e st 
I M P 
D e at h s  
F A S  
I n di vi d u al H a e m at ol o g y R e s ult s 
F A S  
I n di vi d u al Bi o c h e mi str y R e s ult s 
F A S  
C o a g ul ati o n S cr e e ni n g 
F A S  
I n di vi d u al A b n or m al C o a g ul ati o n R e s ult s 
F A S  
I n di vi d u al Uri n al y si s R e s ult s ( di p sti c k) 
F A S  
I n di vi d u al Uri n e Mi cr o s c o p y R e s ult s 
F A S  
I n di vi d u al Vit al Si g n s R e s ult s 
F A S  
I n di vi d u al E C G P ar a m et er s R e s ult s 
F A S  
P h y si c al E x a mi n ati o n 
F A S  
T a bl e s  
 
 A n al y si s P o p ul ati o n s 
F A S  
 P arti ci p a nt Di s p o siti o n 
F A S  
 D e m o gr a p hi c s a n d B a s eli n e C h ar a ct eri sti c s 
F A S  
 S u m m ar y of Pri or M e di c ati o n s 
F A S  
 Pr ot o c ol D e vi ati o n s 
F A S  
 Tr e at m e nt E x p o s ur e 
F A S  
 O v er all S u m m ar y of I n o c ul u m E m er g e nt A d v er s e E v e nt s 
F A S  
 O v er all S u m m ar y of Tr e at m e nt E m er g e nt A d v er s e E v e nt s 
I M P 
 S u m m ar y of I n o c ul u m E m er g e nt A d v er s e E v e nt s b y M e d D R A S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m 
F A S  
 S u m m ar y of Tr e at m e nt E m er g e nt A d v er s e E v e nt s b y M e d D R A S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m 
I M P 
 S u m m ar y of I n o c ul u m E m er g e nt A d v er s e E v e nt s b y M a xi m u m S e verit y  
F A S  
 S u m m ar y of Tr e at m e nt E m er g e nt A d v er s e E v e nt s b y M a xi m u m S e v erit y 
I M P 
 S u m m ar y of I n o c ul u m r el at e d Tr e at m e nt E m er g e nt A d v er s e E v e nt s b y M e d D R A S y st e m Or g a n Cl a s s, Pr ef err e d T er m 
F A S  
 S u m m ar y of I M P R el at e d Tr e at m e nt E m er g e nt A d v er s e E v e nt s b y M e d D R A S y st e m Or g a n Cl a s s, Pr ef err e d T er m 
I M P 
 S u m m ar y of I n o c ul u m R el at e d I E A E s b y S y st e m Or g a n Cl a s s, Pr ef err e d T er m a n d M a xi m u m S e v erit y 
F A S  
 S u m m ar y of I M P R el at e d T E A E s b y S y st e m Or g a n Cl a s s, Pr ef err e d T er m a n d M a xi m u m S e v erit y 
I M P 


Pr ot o c ol N u m b er : M M V _ P yr o n ari di n e _ 2 1 _ 0 1 , 0 6  F e br u ar y 2 0 2 3 
D o c u m e nt st at u s: S A P V er si o n 1. 0  
 
B M S 0 1 -A, v 3 . 0 
St atistic al A n al ysis Pl a n  
Eff ecti v e:  1 7/ S E P / 2 02 0  
C O N FI D E N TI AL  
P a g e 4 9  of 4 9  
 
 I n o c ul u m E m er g e nt S eri o u s A d v er s e E v e nt s b y M e d D R A S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m 
F A S  
 Tr e at m e nt E m er g e nt S eri o u s A d v er s e E v e nt s b y M e d D R A S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m 
I M P 
 O v er all S u m m ar y of I n o c ul u m E m er g e nt A d v er s e E v e nt s of S p e ci al I nt er e st ( A E SI) 
F A S  
 O v er all S u m m ar y of Tr e at m e nt E m er g e nt A d v er s e E v e nt s of S p e ci al I nt er e st ( A E SI) 
I M P 
 I n o c ul u m E m er g e nt A d v er s e E v e nt s of S p e ci al I nt er e st ( A E SI) b y M e d D R A S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m 
F A S  
 Tr e at m e nt E m er g e nt A d v er s e E v e nt s of S p e ci al I nt er e st ( A E SI) b y M e d D R A S y st e m Or g a n Cl a s s a n d Pr ef err e d T er m 
I M P 
 S u m m ar y of C o n c o mit a nt M e di c ati o n s ( F A S) 
F A S  
 S u m m ar y of C o n c o mit a nt M e di c ati o n s (I M P) 
I M P 
 H a e m at ol o g y ( S u m m ar y a n d C h a n g e fr o m I M P B a s eli n e) 
F A S  
 H a e m at ol o g y ( C at e g ori c al) 
F A S  
 Cr o s s - T a b ul ati o n of H a e m at ol o g y A b n or m aliti e s V er s u s I M P B a s eli n e 
F A S  
 Bi o c h e mi str y ( S u m m ar y a n d C h a n g e fr o m I M P B a s eli n e) 
F A S  
 Bi o c h e mi str y ( C at e g ori c al) 
F A S  
 Cr o s s - T a b ul ati o n of Bi o c h e mi str y A b n or m aliti e s V er s u s I M P B a s eli n e 
F A S  
 Li v er F u n cti o n: S u m m ar y of P arti ci p a nt P ot e nti all y Cli ni c all y Si g nifi c a nt A b n or m alit y 
F A S  
 Li v er F u n cti o n: S u m m ar y of P e a k R e a di n g s Si n c e I M P A d mi ni str ati o n 
F A S  
 Li v er F u n cti o n: S u m m ar y of Ti m e t o P e a k R e a di n g s Si n c e I M P A d mi ni str ati o n i n D a y s 
F A S  
 Uri n al y si s ( C at e g ori c al) 
F A S  
 Vit al Si g n s R e s ult s a n d C h a n g e fr o m I M P B a s eli n e 
F A S  
 A b n or m al Vit al Si g n s R e s ult s 
F A S  
 E C G R e s ult s a n d C h a n g e fr o m I M P B a s eli n e 
F A S  
 E C G Q T c C at e g ori c al A n al y s e s 
F A S  
 E C G Cli ni c al A s s e s s m e nt 
F A S  
 
 
 
 


M M V _ P Y R _ 2 1 _ 0 1 S A P _ V 1. 1 _ 0 6 F e b 2 3
Fi n al A u dit R e p ort
2 0 2 3- 0 2- 1 7
Cr e at e d:
2 0 2 3- 0 2- 1 6 ( A u str ali a n C e ntr al St a n d ar d Ti m e ( N ort h er n T errit or y))
B y:
I n g e P at er s o n (i p at er s o n @ s o ut h er n st arr e s e ar c h. c o m)
St at u s:
Si g n e d
Tr a n s a cti o n I D:
C B J C H B C A A B A A k V 7 v A p b v R o R St F h B 2 Q H J 3 Br A w 8 S U 8f h C
" M M V _ P Y R _ 2 1 _ 0 1 S A P _ V 1. 1 _ 0 6 F e b 2 3" Hi st or y
D o c u m e nt cr e at e d b y I n g e P at er s o n (i p at er s o n @ s o ut h er n st arr e s e ar c h. c o m)
2 0 2 3- 0 2- 1 6 - 1 6: 5 0: 5 9 G M T + 9. 5
D o c u m e nt e m ail e d t o M yri a m El G a al o ul ( el g a al o ul m @ m m v. or g) f or si g n at ur e
2 0 2 3- 0 2- 1 6 - 1 6: 5 2: 5 7 G M T + 9. 5
D o c u m e nt e m ail e d t o L o c a di a h K u w a n d a (l k u w a n d a @ s o ut h er n st arr e s e ar c h. c o m) f or si g n at ur e
2 0 2 3- 0 2- 1 6 - 1 6: 5 2: 5 7 G M T + 9. 5
E m ail vi e w e d b y M yri a m El G a al o ul ( el g a al o ul m @ m m v. or g)
2 0 2 3- 0 2- 1 6 - 1 7: 2 9: 1 2 G M T + 9. 5
D o c u m e nt e- si g n e d b y M yri a m El G a al o ul ( el g a al o ul m @ m m v. or g)
Si g ni n g r e a s o n: I a p pr o v e t hi s d o c u m e nt.
Si g n at ur e D at e: 2 0 2 3- 0 2- 1 6 - 1 7: 2 9: 5 1 G M T + 9. 5 - Ti m e S o ur c e: s e r v er
E m ail vi e w e d b y L o c a di a h K u w a n d a (l k u w a n d a @ s o ut h er n st arr e s e ar c h . c o m)
2 0 2 3- 0 2- 1 7 - 1 3: 4 3: 2 2 G M T + 9. 5
D o c u m e nt e- si g n e d b y L o c a di a h K u w a n d a (l k u w a n d a @ s o ut h er n st arr e s e ar c h. c o m)
Si g ni n g r e a s o n: I h a v e a ut h or e d t hi s d o c u m e nt.
Si g n at ur e D at e: 2 0 2 3- 0 2- 1 7 - 1 3: 4 4: 4 5 G M T + 9. 5 - Ti m e S o ur c e: s e r v er
A gr e e m e nt c o m pl et e d.
2 0 2 3- 0 2- 1 7 - 1 3: 4 4: 4 5 G M T + 9. 5