Clinical trial NCT04456634
Conditions: Malaria
Keywords: Saftey Tolerability
Sponsor: Medicines for Malaria Venture
Start date: 2020-09-10; Completion date: 2020-11-17
Enrollment: 8 participants; Allocation: RANDOMIZED; Intervention model: SEQUENTIAL

Brief summary:
Phase 1, single -center study in 2 parts. The study designs for each part are well established for first-in-human studies and are appropriate to assess safety, tolerability and preliminary pharmacokinetics\& pharmacodynamics.

Detailed description:
This is a randomised, single-blinded, placebo-controlled, single centre, phase 1 trial.

Eight healthy males or females, aged between 18-55 years old, who meet all of the inclusion criteria and none of the exclusion criteria, will be enrolled.

The study will be composed of 2 groups to be enrolled sequentially.

• Group 1a (sentinel group): two participants will be randomised single-blinded such that one participant will receive AL+Rux and the other participant will receive AL+placebo.

After review of the safety and tolerability data up to and including Day 8 from Group 1a by the Safety Review Committee (SRC), a decision to proceed with Group 1b will be made.

• Group 1b will be composed of 6 participants, to be randomised single-blinded such that five participants will receive AL+Rux, and one participant will receive AL+placebo.

Arms: arm 1: Oral administration of:

• 20 mg/120 mg artemether-lumefantrine (AL) + 20 mg ruxolitinib phosphate (Rux) arm 2: 20 mg/120 mg artemether-lumefantrine (AL) + Placebo
Interventions: intervention 1: 20 mg/120 mg artemether-lumefantrine (AL) + 20 mg ruxolitinib phosphate (Rux) intervention 2: 20 mg/120 mg artemether-lumefantrine (AL) + Placebo
Intervention descriptions: intervention 1: Rux administered 2 hours after AL administration, twice daily (b.i.d) for 3 consecutive days (6 doses in total). intervention 2: Placebo administered 2 hours after AL administration, twice daily (b.i.d) for 3 consecutive days (6 doses in total).

Protocol documents:
Prot_000.pdf: 
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CLINICAL STUDY PROTOCOL  
A randomised, single blind, placebo controlled, phase 1 trial to evaluate the safety, 
tolerability, pharmacokinetic and pharmacodynamic activity of Ruxolitinib when 
co-administered with artemether-lumefantrine in healthy participants  
 
Protocol Number: 
 
MMV_Ruxolitinib_19_01 
Investigational Product/s: 
 
Ruxolitinib Phosphate (Jakavi®)  
Artemether-lumefantrine (Riamet®) 
Principal Investigator: 
Dr Paul Griffin 
Q-Pharm Pty Ltd 
Level 5, 300C Herston Rd 
Herston Qld 4006, Australia 
Tel: +61 (0)7 3845 3636 
 
Local Sponsor/CRO: 
 
Southern Star Research (SSR) 
1 Merriwa St 
Gordon, NSW 2072 
Australia 
 
Global Sponsor: 
Medicines for Malaria Venture (MMV) 
20, Route de Pré-Bois  
1215 Geneva 15  
Switzerland 
 
Date and Version: 
Version 4.0, 07 September 2020 
 
Confidentiality Statement 
This document contains the confidential information of Medicines for Malaria Venture.  It is 
provided to you as an Investigator or consultant for review by you, your personnel, and the 
applicable Human Research Ethics Committee (HREC). Your acceptance of this document 
constitutes an agreement that you will not disclose the information contained herein to others 
without the prior written consent of Medicines for Malaria Venture, unless it is necessary to obtain 
informed consent from potential study participants. No other use or reproduction is authorized by 
Medicines for Malaria Venture, nor does Medicines for Malaria Venture assume any 
responsibility for unauthorized use of this protocol. 
 
 


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1 PROTOCOL SUMMARY 
1.1 
PROTOCOL SYNOPSIS 
Protocol Title 
A randomised, single blind, placebo controlled, phase 1 trial to evaluate the 
safety, tolerability, pharmacokinetic and pharmacodynamic activity of 
Ruxolitinib when co-administered with artemether-lumefantrine in healthy 
participants. 
IND Number 
N/A 
Protocol Number 
MMV_Ruxolitinib_19_01 
Local Sponsor 
Southern Star Research Pty Ltd 
Global Sponsor 
Medicines for Malaria Venture  
Principal Investigator 
Dr Paul Griffin, Q-Pharm Pty Ltd 
Phase of Development 
Phase 1 
Number of Study Sites 
1 
Treatment Groups 
Group 1a (sentinel group):  
Oral administration of:  
• 
20 mg/120 mg artemether-lumefantrine (AL) +  20 mg ruxolitinib 
phosphate (Rux) or  
• 
20 mg/120 mg AL + placebo;  
Rux or placebo administered 2 hours after AL administration, twice daily 
(b.i.d) for 3 consecutive days (6 doses in total). 
The Principal Investigator, Sponsor Medical Director and study Medical 
Monitor (at a minimum) will review safety and tolerability data up to and 
including Day 8 from Group 1a before deciding if Group 1b can proceed. The 
dose of 20 mg Rux will not be altered. 
Group 1b: Oral administration of: 
• 
20 mg/120 mg AL + 20 mg Rux or 20 mg/120 mg AL + placebo; 
• 
Rux or placebo administered 2 hours after AL administration, twice 
daily (b.i.d) for 3 consecutive days (6 doses in total) 
Participant 
Population 
Healthy male and female adult volunteers that meet all eligibility criteria will 
be enrolled: 
2 groups each to be enrolled sequentially. Group 1a will be a sentinel group 
of 2 participants, and Group 1b will be composed of 6 participants;   
randomised (single blinded; treatment allocation concealed to participants but 
not Investigator) to be administered either AL + Ruxolitinib or AL + placebo.  
Investigational 
Medicinal 
Products 
(IMP) 
• Artemether-lumefantrine (AL), 20mg/120mg tablets administered orally 
2 hours prior to:  
• Ruxolitinib phosphate (Rux), 20 mg tablet administered orally or placebo 
capsule administered orally.  
Duration of Study 
Approximately 2 months for each participant - screening period of up to 28 
days and 28 days on-study, with total of up to 56 days. 
Objectives 
Primary Objective: 
To assess the safety and tolerability of 3-day b.i.d dosing of AL+Rux and 
AL+placebo.  
Secondary Objectives: 


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1. To assess the effect of AL+Rux and AL+placebo on pSTAT3 inhibition. 
2. To characterise the PK profiles of artemether and its major metabolite 
dihydroartemisinin [DHA], lumefantrine, and Rux.  
3. To assess additional safety variables (tympanic body temperature and 
respiration rate) 
Endpoints 
Primary Endpoint: 
1. Incidence, severity, and relationship of observed and self-reported AEs 
up to 28 days after AL+Rux and AL+placebo administration in all 
participants by treatment regimen.  
2. Clinical laboratory evaluations including haematology, biochemistry and 
urinalysis.  
3. Vital signs (blood pressure and heart rate).  
4. 12-lead standard ECG: QT, QTcB and QTcF; HR, PR, QRS. 
Secondary Endpoints: 
1. pSTAT3 inhibition ex-vivo on whole blood cells by: 
• 
pSTAT3 levels pre- and post- AL+Rux administration intra participant 
and, 
• 
pSTAT3 levels per treatment regimen post AL+Rux and post 
AL+placebo administration 
2. PK parameters of artemether, DHA, lumefantrine, and Rux using non-
compartmental methods: AUClast, AUC0-, AUC0-8, AUC60-72, Cmax (first 
and last dose), tmax (first and last dose), elimination half-life (t½), tlag, C168h 
(for lumefantrine only), CL/F, Vz/F and λz in all participants.  
3. Safety variables tympanic body temperature (°Celcius) and respiratory 
rate.  
Study Description 
This is a randomised, single-blinded, placebo-controlled, single centre, phase 
1 trial. Eight healthy males or females, aged between 18-55 years old, who 
meet all of the inclusion criteria and none of the exclusion criteria, will be 
enrolled. 
The study will be composed of 2 groups to be enrolled sequentially.  
• Group 1a (sentinel group): two participants will be randomised single-
blinded such that one participant will receive AL+Rux and the other 
participant will receive AL+placebo.  
After review of the safety and tolerability data up to and including Day 8 from 
Group 1a by the Principal Investigator, the Sponsor Medical Director and the 
study Medical Monitor (at a minimum), a decision to proceed with Group 1b 
will be made. 
• Group 1b will be composed of 6 participants, to be randomised single-
blinded such that five participants will receive AL+Rux, and one participant 
will receive AL+placebo.  
Participants will be admitted to the clinical trial unit on Day -1 and will be 
administered an oral dose of AL+Rux or AL+placebo separated by 2 hours 
(with Rux or placebo administered 2 hours after AL) b.i.d commencing on 
Day 1 (AL at t = 0, 8, 24, 36, 48 and 60 hours). Participants will be confined 
to the clinical trial unit for at least 78 hours after first dose IMP administration 
for blood sampling for PK and PD (pSTAT3) and close safety and tolerability 
monitoring. After discharge from the clinical trial unit, participants will be 
followed via outpatient care visits for safety assessments, pSTAT sampling 
(day 8) and PK blood sampling up to day 29.  
Sample Size 
Eight healthy males or females, aged between 18 and 55 years old, who meet 
all of the inclusion criteria and none of the exclusion criteria will be enrolled. 
As the combination of Rux+AL has not been tested in humans before, a first-
in-human approach of eight participants (with ratio active to placebo of 3:1) 


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has been selected. Additional reserve volunteers will be consented and 
screened to be available on Day 1 in the event of withdrawal of participants 
prior to first-dose IMP administration.  
Inclusion Criteria 
Potential participants must fulfil all of the following inclusion criteria to be 
eligible to participate in the study and throughout the study: 
1. Male or female (non-pregnant, non-lactating) aged 18 to 55 years 
inclusive. 
2. Contactable and available for the duration of the trial and for up to two 
weeks following the EOS visit. 
3. 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) at 
Screening and Day -1. 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 
4. Certified as healthy by a comprehensive clinical assessment (detailed 
medical history, full physical examination and special investigations). 
5. Vital signs 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. ECG parameters for both males and females: QT ≤ 500 msec, QTcF  
≤450 msec, QTcB ≤450 msec; PR interval ≤210 msec.  
7. Heterosexual female participants of childbearing potential who have, or 
may have, male sexual partners during the course of the study should be 
using an insertable (implant or IUD), injectable, transdermal or 
combination oral contraceptive approved by the TGA combined with a 
barrier contraceptive from the time of informed consent until 30 days 
after EOS. Abstinent female participants must agree to start a double 
method if they start a sexual relationship with a male during the trial 
until 30 days after EOS. Female participants must not be planning in 
vitro fertilisation within the required contraception period. 
Women of non-childbearing potential who will not require 
contraception during the trial are defined as: surgically sterile (tubal 
ligation is not considered surgically sterile), post-menopausal 
(spontaneous 
amenorrhoea 
for 
≥12 
months, 
or 
spontaneous 
amenorrhoea for 6-12 months and follicle-stimulating hormone (FSH) 
≥40 IU/mL; either should be together with the absence of oral 
contraceptive use for >12 months). 
Male participants who have, or may have female sexual partners during 
the course of the study must agree to use a double method of 
contraception including condom plus diaphragm, or condom plus stable 
insertable (implant or IUD), injectable, transdermal or combination oral 
contraceptive by the female partner, from the time of informed consent 
through to 90 days after EOS. Abstinent male participants must agree to 
start a double method if they begin a sexual relationship with a female 
during the trial, and through to 90 days after EOS. Male participants with 
female partners that are surgically sterile or post-menopausal, or male 
participants 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.  
In addition, all male participants must agree to not donate sperm for at 
least 90 days after EOS.  
Regulations 
8. Completion of the written informed consent process prior to 
undertaking any trial-related procedure.  


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9. Must be willing and able to communicate and participate in the whole 
trial. 
10. Agree to adhere to Lifestyle Considerations (see Section 4.3.3) 
throughout trial duration and be willing to consume 250 mL full-fat 
milk with each dose of AL. 
Exclusion Criteria 
If any of the following exclusion criteria apply, the potential participant will 
not be permitted to participate or remain in the study: 
Medical history and clinical status 
1. Known hypersensitivity to ruxolitinib, artesunate or any of its excipients, 
artemether, 
lumefantrine 
or 
other 
artemisinin 
derivatives, 
proguanil/atovaquone, primaquine, or 4-aminoquinolines. 
2. Haematology, 
biochemistry 
or 
urinalysis 
results 
that 
are 
abnormal/outside of the laboratory normal reference ranges AND are 
either: 
• Considered clinically signficant by the Principal Investigator or 
delegate; OR 
• Considered not clinically significant by the Principal Investigator or 
delegate BUT ARE ALSO outside of Sponsor-approved clinically 
acceptable laboratory ranges in Appendix 1.  
NOTE: Participants are not excluded if abnormal/out of laboratory 
normal reference range results are considered not clinically significant 
by the Principal Investigator or delegate AND are within the ranges 
specified in Appendix 1. 
3. Platelets < 200x109/L at Screening or prior to IMP administration is 
exclusionary. One re-test is permitted if original test result does not 
reflect the assumed medical status of the individual.  
4. Participation in any other investigational product trial within 5 half-lives 
or 12 weeks preceding IMP administration, whichever is longer, or in the 
exclusion period of a previous trial according to applicable regulations. 
5. Symptomatic postural hypotension at screening and pre-first dose of IMP 
on Day 1 (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. History or presence of diagnosed (by an allergist/immunologist) or 
treated (by a physician) food or known drug allergies, or any history of 
anaphylaxis or other severe allergic reactions including face, mouth, or 
throat swelling or any difficulty breathing. 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 is not an 
exclusion criterion. 
8. Presence of current or suspected serious chronic diseases such as cardiac 
or autoimmune disease (HIV or other immuno-deficiencies), insulin-
dependent and non-insulin-dependent diabetes, progressive neurological 
disease, severe malnutrition, acute or progressive hepatic disease, acute 
or progressive renal disease, porphyria, psoriasis, rheumatoid arthritis, 
asthma (excluding childhood asthma), epilepsy, or obsessive-compulsive 
disorder. 
9. History of malignancy of any organ system (other than localised and 
considered cured basal cell carcinoma of the skin or in situ cervical 
cancer), treated or untreated, within five years of screening, regardless of 
whether there is no 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 


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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. 
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 (BDI-II) will be used as a validated tool for the 
assessment of depression at screening. In addition to the conditions listed 
above, individuals 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 individuals will be 
referred to a general practitioner or medical specialist as appropriate. 
Individuals 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 individual 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. Acute illness within the 4 weeks prior to screening and prior to IMP 
administration.   
15. Significant inter-current disease of any type, in particular liver, renal, 
cardiac, pulmonary, neurologic, rheumatologic, or autoimmune disease 
by history, physical examination, and/or laboratory studies including 
urinalysis. 
16. Individual has a clinically significant disease or any condition or disease 
that might affect drug absorption, distribution or excretion (e.g. 
gastrectomy, cholecystectomy, diarrhoea) or known lactose/dairy 
intolerance. 
17. Participation in any research trial involving blood sampling (more than 
300 mL/unit of blood) within one month prior to IMP administration, or 
blood donation to Australian Red Cross Blood Service (Blood Service) 
or other blood bank during the 8 weeks prior to IMP administration. 
18. Medical requirement for intravenous immunoglobulin or blood 
transfusions. 
19. 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. 
20. Any individual who has ever smoked >1 pack of cigarettes per day for 
>10 years, or who currently (within 14 days prior to Screening or prior 
to IMP administration smokes >5 cigarettes/day. 
21. Female who is breastfeeding.   
22. Any vaccination within the last 28 days prior to screening or prior to IMP 
administration. 
23. Prior to screening or IMP administration: any systemic or inhaled 
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 topical, nasal or ophthalmic 
corticosteroids within the past 2 weeks. 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. 
24. Use of antidepressant medication in the past 12 months prior to screening 
or prior to IMP administration. 
25. Use of any other medication except contraceptives (including herbal, 
vitamin supplement, OTC or prescription) within 14 days or five half-


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lives (whichever is longest) prior to IMP administration. Participants are 
requested to refrain from taking non-approved concomitant medications 
from recruitment until the conclusion of the trial. 
26. Cardiac/QT risk: 
• Family history of sudden death or of congenital prolongation of the 
QT, QTcF, or  QTcB interval or known congenital prolongation of the 
QT, QTcF, or  QTcB interval or any clinical condition known to 
prolong the QT, QTcF, or  QTcB interval. 
• History of symptomatic cardiac arrhythmias or with clinically relevant 
bradycardia. 
• Electrolyte disturbances, particularly hypokalaemia, hypocalcaemia, 
or hypomagnesaemia. 
• ECG abnormalities in the standard 12-lead ECG (at screening or prior 
to IMP administration) which in the opinion of an Investigator is 
clinically relevant or will interfere with the ECG analyses.  
General conditions 
27. 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.  
28. Any individual for whom study participation would pose an additional 
safety risk as assessed by the Principal Investigator.  
29. Any individual who is an Investigator, research assistant, pharmacist, 
trial coordinator, or other staff thereof, directly involved in conducting 
the trial. 
30. Any individual without good peripheral venous access. 
Biological status 
31. 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). 
32. Recent herpes zoster infection (within the previous 6 months) as 
determined by clinical history. 
33. Positive result for M. tuberculosis infection by QuantiFERON-TB Gold 
assay. 
34. Positive urine drug test for any drug listed in Section 7.4.5. Any 
individual testing positive for acetaminophen (paracetamol) at screening 
and/or pre-dose  may still be eligible for trial participation at the 
discretion of the Principal Investigator or delegate. 
35. Positive alcohol breath test. 
Statistical Analysis 
• Four analysis datasets will be used for study analyses: Full analysis set 
(FAS), Safety Set, PK set and pSTAT3 set.  
• The FAS will be used to list and summarise subject disposition, 
demographics and baseline characteristics, protocol deviations and 
treatment exposure.  
• Safety and tolerability will be assessed by clinical review of all AEs, vital 
signs, 12-lead ECG, haematology, biochemistry, urinalysis and physical 
examination results. All descriptive statistics will be evaluated using the 
Safety Set.  
• PK parameters of artemether, DHA, lumefantrine and Rux will be 
estimated using non-compartmental methods from plasma concentration-
time data. For calculation of descriptive statistics of plasma 
concentrations, values below the lower limit of quantitation will be set to 
zero. PK analyses will be conducted using the PK Set. 
• Levels of pSTAT3 will be expressed as a percentage and summarised by 
percentage inhibition pre- and post- IMP administration and as a change 
in percentage between pre- and post- IMP administration. The following 
analyses will be conducted: within-participant comparison of each 
participant before AL+Rux or AL+placebo administration, and 12 hours 


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after first administration of AL; between-participants comparison of 
participants who receive AL+Rux and participants who receive 
AL+placebo. Analyses of pSTAT3 levels will be conducted using the 
pSTAT3 Set.  
 
 


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1.2 
SCHEMA 
 
Figure 1: Schema 
  
 


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1.3 
SCHEDULE OF ACTIVITIES 
Table 1: Schedule of Activities - Overview 
Part 1 
Activity/Procedure 
Screening/Eligibility/ Safety 
 
Treatment and 
Confinement  
 
Outpatient 
Monitoring 
 
EOS/ET 
 
 
Day-28 to Day -1 
Day -1a 
Day 1 to Day 4a 
Day 5 to Day 29a 
Day 29a 
Overnight fast of ≥8 h required  
X 
X 
X  
X  Days 8 & 15 
X 
Informed consent, BDI-II & demographics 
X 
 
 
 
 
Medical history, eligibility & prior/concomitant 
medications; recreational drug, alcohol and 
tobacco use (history and current) 
X 
X 
X pre-dose 
 
 
Confinement at clinical unit 
 
X 
X 
 
 
Discharge from clinical unit 
 
 
X 78 h post-first dose IMP 
 
 
Outpatient visit to clinical unit 
X 
X 
 
X 
X 
Urine drug screen & alcohol breath test 
X 
X  
 
 
 
Full physical examination 
X 
 
 
 
X 
Abbreviated physical examination 
 
X  
 
 
 
Symptom-directed physical examination if 
clinically indicated 
 
 
X during & prior to 
discharge 
X 
 
Vital signs  (BP & HR) 
Xb 
Xb  
Xc  
Xd 
Xd 
Tympanic Body Temperature (°C) & RR 
(supine) 
X 
X 
X 4 h & every 24 h post-first 
dose IMP 
X 
X 
Height (cm) and weight (kg) 
X 
X weight only 
 
 
 
12-lead ECG 
X 
 
X pre-dose, 24, 48, 72 h 
post-first dose IMP 
X  Days 8 & 15 
X 
Urinalysis 
X 
Xe 
X 24, 48, 72 h post-first dose 
IMP  
 
X  Days 8 & 15 
X 
Intravenous cannulation 
 
X or Day 1 
X or Day -1 
 
 
Haematology  
X  
 
Xe 
X 24, 48, 72 h post-first dose 
IMP 
 
X  Days 8 & 15 
X 
Fasting Biochemistry (may include serum  
β-hCG, 
FSH 
[post-menopausal 
female 
participants only], and lipids) 
X  incl. β-hCG (all females), 
FSH, & lipids, blood group, 
Rh(D) (fasting) 
Xe  
X 24, 48, 72 h post-first dose 
IMP  
X  Days 8 & 15 
X incl. 
β-hCG (WOCBP only)  


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Part 1 
Activity/Procedure 
Screening/Eligibility/ Safety 
 
Treatment and 
Confinement  
 
Outpatient 
Monitoring 
 
EOS/ET 
 
 
Day-28 to Day -1 
Day -1a 
Day 1 to Day 4a 
Day 5 to Day 29a 
Day 29a 
Coagulation profile 
X 
 
 
 
 
Serology  
 
X  
 
 
 
 
QuantiFERON-TB Gold assay  
X 
 
 
 
 
Urine β-hCG pregnancy test (WOCBP only) 
 
X  
 
 
 
AEs & concomitant medicationsf 
 
 
X f 
X 
X 
AL and Rux PK blood sampling 
 
 
Xg 
Xh 
X 
pSTAT3 blood sampling 
 
 
Xi 
Xi Day 8 (168h) 
 
IMP administration 
 
 
Xj 
 
 
Abbreviations: AE=adverse event; AL=artemether/lumefantrine (Riamet); BDI-II=Beck Depression Inventory; BP = blood pressure;  ECG=electrocardiogram;  
FSH=follicle stimulating hormone; G6PD=glucose-6-phosphate dehydrogenase; hCG=human chorionic gonadotropin; HR = heart rate;  IMP=investigational medical 
product; inc.=including; PK=pharmacokinetics; pSTAT3=phospho-signal transducer and activator of transcription 3; RR = respiratory rate; Rux=ruxolitinib 
(Jakavi®); Safety labs=blood collection for haematology and biochemistry; TB=tuberculosis; WOCBP=women of child bearing potential. 
 
a. Day 1 is defined as the day of first dose of IMP (AL at t = 0). Participants will be admitted to the clinical unit on Day -1 for confinement and administration 
of IMP on Day 1, and will be confined to the unit for at least 78 hours post-first dose IMP. Outpatient visits are scheduled on Days 8 (t = 168h), 11 (t = 
240h), 15 (t = 336h), 21 (t = 480h), 24 (t = 552h) and 27 (t = 624h) with EOS on Day 29 (t = 672h). Visit windows for all outpatient visits are as per the 
PK sampling time windows for those timepoints as described in Section 6.2.  
b. Vital signs (blood pressure (BP; systolic and diastolic) and heart rate (HR) at screening and on Day -1: blood pressure and heart rate measured supine 
after resting in supine position for at least 5 minutes and standing within 2-3 minutes when changing from supine to standing. 
c. On Day 1, BP and HR measured pre-first dose (measured supine and standing as per footnote b), and then after participant supine for at least 5 minutes 
at 4 and 8 hours post-first dose IMP during confinement and then every 8 hours. Pre-first dose results must satisfy relevant inclusion/exclusion criteria.  
d. Vital signs (BP and HR) conducted after participant has rested in supine position for at least 5 minutes. 


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e. Haematology biochemistry and urinalysis results from samples collected on Day -1 must be available for safety review on Day 1 prior to first-dose IMP 
(AL at t = 0) and must satisfy relevant inclusion/exclusion criteria. 
f. Untoward medical events reported between the time of informed consent and first dose of IMP will be recorded as medical history, and must satisfy 
relevant inclusion/exclusion criteria. Adverse events will be recorded from time of first dose of IMP (AL at t = 0).  
g. During confinement, PK blood sampling to be taken at timepoints pre-dose, then at t = 1, 2, 3, 4, 5, 6, 8, 12, 24, 36, 48, 60, 62, 64, 72 and 78 hours post 
commencement of IMP dosing.  When sampling coincides with IMP administration, blood samples must be taken prior to IMP. See Section 6.2 for 
permitted time windows. 
h. During outpatient visits, PK blood sampling to be taken on Days 8, 11, 15, 21, 24, 27 and 29 (EOS) at t = 168, 240, 336, 480, 552, 624 and 672 hours 
post commencement of IMP dosing respectively. See Section 6.2 for permitted time windows. 
i. 
During confinement, pSTAT3 blood sampling to be taken at timepoints pre-dose, then t = 1, 2, 3, 4, 5, 6, 8, and 12 hours post commencement of IMP 
dosing.When sampling coincides with IMP administration, blood samples must be taken prior to IMP. A pSTAT3 blood sample will also be taken at t = 
168 hours post-first dose IMP (AL at t = 0) on Day 8. See Section 6.2 for permitted time windows. 
j. Randomised (on Day -1 or Day 1) to be administered either AL+Rux or AL+placebo from Day 1. AL dosing (with 250 mL full-fat milk)  at t=0, 8, 24, 
36, 48 and 60 hours (see Section 5.1.1 for food and other restrictions). Rux/placebo administered with 250 mL water 2 hours after AL at t = 2, 10, 26, 
38, 50 and 62 hours (see Sections 5.1.2 and 5.1.3 for food and other restrictions). All doses to be administered under supervision and at the scheduled 
timepoints. NOTE: Results of safety laboratory testing (haematology, biochemistry, urinalysis) taken during the Safety/Eligibility visit must be available 
for review by the Principal Investigator or delegate prior to IMP administration on Day 1. All pre-first dose assessment results must satisfy relevant 
inclusion/exclusion criteria.  
 
NOTE: This table provides an overview only for activities required from Day 1. See Table 2 for detailed Schedule of Activities including all 
scheduled timepoints for Day 1 to Day 4 Confinement and IMP Treatment; see Table 3 for detailed Schedule of Activities for Outpatient Visits 
after discharge from the clinical unit including EOS/ET.  
 
 
 


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Table 2: Schedule of Activities – Detailed Day 1 to Day 4 Confinement and IMP Treatment 
Daya 
D1a 
D2a 
D3a 
D4a 
Time 
(hour)a 
Pre
-
dos
e 
0H 
1H 
2H 
3H 
4H 
5H 
6H 
8H 
10
H 
12
H 
16
H 
24
H 
26
H 
32
H 
36
H 
38
H 
40
H 
48
H 
50
H 
56
H 
60
H 
62
H 
64
H 
72
H 
78
H 
Confinement 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
 
Discharge 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
X 
Randomizati
on 
Xb 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
IMPc 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
IMP - AL 
 
X 
 
 
 
 
 
 
X 
 
 
 
X 
 
 
X 
 
 
X 
 
 
X 
 
 
 
 
IMP – 
Rux/placebo 
 
 
 
X 
 
 
 
 
 
X 
 
 
 
X 
 
 
X 
 
 
X 
 
 
X 
 
 
 
Safetyd 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Physical 
exam sympt. 
 
Xe 
Xe 
Vital signsf 
(BP & HR)  
Xf 
 
 
 
 
X 
 
 
X 
 
 
X 
X 
 
X 
 
 
X 
X 
 
X 
 
 
X 
X 
 
Tympanic 
body 
temperature 
(°C) and RR 
Xg 
 
 
 
 
Xg 
 
 
 
 
 
 
Xg 
 
 
 
 
 
Xg 
 
 
 
 
 
Xg 
 
Standard 12-
lead ECG 
Xh 
 
 
 
 
 
 
 
 
 
 
 
Xh 
 
 
 
 
 
Xh 
 
 
 
 
 
Xh 
 
Biochemistry 
Haematology 
 
 
 
 
 
 
 
 
 
 
 
 
Xi 
 
 
 
 
 
Xi 
 
 
 
 
 
Xi 
 
Urinalysis 
 
 
 
 
 
 
 
 
 
 
 
 
Xi 
 
 
 
 
 
Xi 
 
 
 
 
 
Xi 
 
Record AE & 
Concomitant 
medicationsj 
 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
X 
PK 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
PK blood  
samplesk 
X 
 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
X 
 
 
X 
 
 
X 
 
 
X 
X 
X 
X 
X 


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Daya 
D1a 
D2a 
D3a 
D4a 
Time 
(hour)a 
Pre
-
dos
e 
0H 
1H 
2H 
3H 
4H 
5H 
6H 
8H 
10
H 
12
H 
16
H 
24
H 
26
H 
32
H 
36
H 
38
H 
40
H 
48
H 
50
H 
56
H 
60
H 
62
H 
64
H 
72
H 
78
H 
PD 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
pSTAT3 
samplel 
X 
 
X 
X 
X 
X 
X 
X 
X 
 
X 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
ABBREVIATIONS: abbrev. = abbreviated; AE = adverse event; AL = artemether/lumefantrine (Riamet®); BP = blood pressure; ECG = electrocardiogram; exam 
= examination; hCG = human chorionic gonadotropin; HR = heart rate; IMP = investigational medicinal product; PD = pharmacodynamics; PK = 
pharmacokinetics; RR = respiratory rate; Rux = ruxolitinib (Jakavi®); sympt. = symptom directed; WOCBP = woman of child bearing potential. 
a. Time (hour) is defined as the time in hours after the first dose of IMP (AL at t = 0 on Day 1). NOTE: Study days are numbered by convention per 24 hour 
period from t = 0. The actual day of some timepoints in practice may vary depending on the actual time of t = 0 on Day 1. Participants will be admitted to 
the clinical unit on Day -1 for treatment and confinement, and will not be discharged from the clinical unit until at least 78 hours post-first dose IMP. 
b. If not performed on Day -1, participants will be randomized (single-blinded – participant is unaware of treatment allocation) to be administered either 
AL+Rux or AL+placebo. 
c. IMPs will be administered under direct observation at the scheduled timepoints (exact timing to be respected): AL (20 mg artemether, 120 mg lumefantrine 
per tablet): 4 tablets administered orally with 250 mL full-fat milk at t = 0, 8, 24, 36, 48 and 60 hours (total 24 tablets). Rux (20 mg ruxolitinib per tablet) or 
placebo capsule: 1 tablet/capsule administered orally with 250 mL water 2 hours after each AL administration at t = 2, 10, 26, 38, 50 and 62 hours (total 6 
tablets/capsules). Participants will be blindfolded for Rux/placebo dosing and prevented from witnessing each other’s Rux/placebo dosing. Participants should 
swallow IMP whole without biting or chewing. No food permitted 30 minutes prior to AL dosing, between AL and Rux dosing or 1 hour after Rux dosing. 
If dosing coincides with other assessments, participant should remain seated for 10 minutes post-dose, then remain semi-recumbent for at least 1 hour. NOTE: 
Results from biochemistry/haematology/urinalysis taken at eligibility/safety visit must be available and reviewed by the Principal Investigator or delegate 
prior to first dosing with IMP (AL at t = 0). On Day 1, participants will have fasted for at least 8 hours overnight before breakfast prior to first-dose IMP at t 
= 0. All pre-first dose assessment results must satisfy relevant inclusion/exclusion criteria. 
d. Refer to Safety Section 7 for detailed safety investigations.  
e. Symptom-directed physical examination when required if clinically indicated during confinement. Any pre-first dose results must satisfy relevant 
inclusion/exclusion criteria. Prior to participant discharge, clinical unit staff must clarify with the Principal Investigator or delegate if a symptom directed 
physical examination is required.  
f. Vital signs during confinement (systolic and diastolic blood pressure [mmHg] and heart rate [beats per minute): NOTE: pre-dose within 60 minutes prior to 
first-dose IMP (AL at t = 0): blood pressure measured after participant supine for at least 5 minutes and then again in standing position within 2-3 minutes 
of standing (pre-first dose results must satisfy relevant inclusion/exclusion criteria). Then vital signs measured within 15 minutes prior to each subsequent 
timepoint during confinement after participant supine for at least 5 minutes at: 4 and 8 hours post-first dose IMP, and then every 8 hours during confinement.  


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g. Respiratory rate (RR: breaths per minute) and tympanic body temperature (ºCelcius) will be measured within 60 minutes prior to first-dose IMP (AL at t = 
0; pre-first dose results must satisfy any relevant inclusion/exclusion criteria), and then 15 minutes prior to the subsequent timepoints during confinement. 
RR will be measured after participant is supine for at least 5 minutes.  
h. Standard single 12-lead ECG recorded after supine for at least 5 minutes within 60 minutes prior to first-dose IMP (AL at t = 0; pre-first dose results must 
satisfy relevant inclusion/exclusion criteria), then within 15 minutes prior to subsequent timepoints at 24, 48 and 72 hours post-first dose IMP (AL at t = 0).  
i. Safety laboratory samples (haematology, biochemistry, urinalysis) will be collected 24, 48 and 72 hours post-first dose IMP (within 60 minutes prior to each 
timepoint – must be collected prior to breakfast). Safety biochemistry laboratory sampling requires fasting for at least 8 hours. 
j. Untoward medical events reported before the first dose of IMP will be recorded as medical history and must satisfy relevant inclusion/exclusion criteria. 
Adverse events and any concomitant medications will be recorded from time of first dose of IMP. AE/concomitant medication timepoint checks: ± 10 minutes 
for Day 1, then ± 20 minutes for remainder of confinement. AEs/concomitant medications may be reported/ recorded at any time during confinement. See 
Section 7.6 for further information including SAE/AESI reporting. See Section 4.3.8 for toxicity/stopping rules.  
k. PK blood samples will be collected at the timepoints indicated. For timepoints coinciding with IMP administration, blood sampling must occur prior to IMP 
administration. The exact timing for IMP administration must be respected. See Section 6.2 for permitted time windows. 
l. pSTAT3 blood samples will be collected at the timepoints indicated. For timepoints coinciding with IMP administration, blood sampling must occur prior to 
IMP administration. See Section 6.2 for permitted time windows. 
NOTE:  
• 
When several procedures are scheduled to take place at the same timepoint, the following order is recommended where applicable: urinalysis, 
ECG, vital signs, blood sampling, drug administration. 
• 
Date/time of all assessments/activities must be recorded in source document.  
• 
Pre-dose assessments will be conducted within 60 minutes prior to the first dose of IMP (AL at t = 0).  
 
 
 
 


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Table 3: Schedule of Activities – Detailed Days 8 to EOS Outpatient Visits 
Day (D) 
D8 
D11 
D15 
D21 
D24 
D27 
EOS D29 
(or ET) 
 
Time 
(hour)a 
168H 
 120 min 
240H 
24h 
336H 
24h 
480H 
24h 
552H 
24h 
624H 
24h 
672H 
 24h 
Outpatient visit at clinical site 
X 
X 
X 
X 
X 
X 
X 
Safetyb 
 
 
 
 
 
 
 
Physical exam: Full 
 
 
 
 
 
 
X 
Physical exam:Symptom-directed 
Xc 
Xc 
Xc 
Xc 
Xc 
Xc 
 
Vital signs (supine)d 
X 
X 
X 
X 
X 
X 
X 
Tympanic body temperature (°C) and RR d 
X 
X 
X 
X 
X 
X 
X 
Standard single 12-lead ECG 
Xe 
 
Xe 
 
 
 
Xe 
Biochemistry, Haematology 
Xf 
 
Xf 
 
 
 
Xf 
Urinalysis 
X 
 
X 
 
 
 
X 
Serum -hCG  (WOCBP only) 
 
 
 
 
 
 
X 
Record AE and concomitant medicationsg 
X 
X 
X 
X 
X 
X 
X 
PK 
 
 
 
 
 
 
 
PK blood  samplesh 
X 
X 
X 
X 
X 
X 
X 
PD 
 
 
 
 
 
 
 
pSTAT3 sample 
Xi 
 
 
 
 
 
 
ABBREVIATIONS: AE = adverse event; ECG = electrocardiogram; exam = examination; ET = early termination visit; hCG = human chorionic gonadotropin; PD 
= pharmacodynamic; PK = pharmacokinetic; WOCBP = woman of child bearing potential. 
a. Time (hour) is defined as the time in hours after the first dose of IMP at t=0 on Day 1.  
b. Refer to Safety Section 7 for detailed safety investigations. All assessments to be conducted within the same time window permitted for PK sampling (Section 
6.2).  
c. Symptom-directed physical examination when required if clinically indicated. Clinical unit staff must clarify with the Principal Investigator or delegate if a 
symptom directed physical examination is required.  
d. Vital signs (systolic and diastolic blood pressure [mmHg] and heart rate [beats per minute]) and respiratory rate (breaths per minutes) to be measured after 
participant supine for at least 5 minutes.  
e. Standard single 12-lead ECG recorded after participant supine for at least 5 minutes.  
f. Safety biochemistry laboratory sampling on Days 8, 15 and EOS require fasting for at least 8 hours.  


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g. All adverse events that occur and/or concomitant medications that are administered after discharge from the clinical unit and until EOS will be recorded. See 
Section 7.6 for further information including SAE/AESI reporting.  
h. The exact timing for PK sampling must be respected. See Section 6.2 for permitted time windows. NOTE: permitted outpatient visit windows are with 
respect to the scheduled PK sampling timepoints in hours post-first dose IMP at t = 0.  
i. pSTAT3 sampling: see Section 6.2 for permitted time windows. 
NOTE:  
• 
When several items take place at the same time, the following order is recommended where applicable: urinalysis, ECG, vital signs, blood 
sampling. 
• 
Date/time of all assessments/activities must be recorded in source document.  
• 
Time windows for all outpatient assessments are as per PK/PD sampling time windows (Section 6.2).  
 
 


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TABLE OF CONTENTS 
1 
PROTOCOL SUMMARY .......................................................................... 2 
1.1 
PROTOCOL SYNOPSIS................................................................................... 2 
1.2 
SCHEMA .............................................................................................................. 9 
1.3 
SCHEDULE OF ACTIVITIES ................................................................................. 10 
2 
INTRODUCTION AND RATIONALE................................................... 29 
2.1 
DISEASE BACKGROUND .................................................................................... 29 
2.2 
INVESTIGATIONAL MEDICINAL PRODUCTS ....................................................... 30 
2.2.1 
Ruxolitinib (Jakavi®) ............................................................................... 30 
2.2.1.1 
Clinical Studies ................................................................................ 30 
2.2.2 
Riamet® - Artemether/lumefantrine (AL) .................................................. 31 
2.2.3 
Clinical Safety and Risk Assessment ........................................................ 31 
2.2.3.1 
Ruxolitinib ....................................................................................... 31 
2.2.3.2 
Riamet® - Artemether/lumefantrine (AL) ........................................ 32 
2.3 
RATIONALE ....................................................................................................... 32 
2.3.1 
Rationale for the Study ............................................................................ 32 
2.3.2 
Rationale for the Dose ............................................................................. 33 
3 
STUDY OBJECTIVES AND ENDPOINTS............................................ 34 
4 
INVESTIGATIONAL PLAN ................................................................... 35 
4.1 
DESCRIPTION OF OVERALL STUDY DESIGN AND PLAN ..................................... 35 
4.1.1 
Group 1a (sentinel group) ....................................................................... 35 
4.1.2 
Group 1b .................................................................................................. 35 
4.2 
DISCUSSION OF STUDY DESIGN INCLUDING CHOICE OF CONTROL GROUPS ...... 35 
4.3 
SELECTION OF STUDY POPULATION .................................................................. 36 
4.3.1 
Inclusion Criteria ..................................................................................... 36 
4.3.2 
Exclusion Criteria .................................................................................... 37 
4.3.3 
Lifestyle Considerations .......................................................................... 41 
4.3.4 
Screen Failures ........................................................................................ 41 
4.3.5 
Withdrawal of Participants ...................................................................... 42 
4.3.5.1 
Replacement of Withdrawn/Discontinued Participants ................... 42 
4.3.6 
Lost to Follow-Up .................................................................................... 43 
4.3.7 
Study Discontinuation .............................................................................. 43 
4.3.8 
Trial Intervention/Treatment Discontinuation – Toxicity Rules .............. 43 
4.3.8.1 
Individual Toxicity Rules ................................................................ 43 
4.3.8.2 
Group Toxicity Rules ...................................................................... 44 
4.3.8.3 
Special Individual Toxicity Rules ................................................... 44 
4.3.8.4 
Special Group Toxicity Rules .......................................................... 44 
4.3.9 
Emergency Unblinding of Study Participants ......................................... 44 
5 
TRIAL INTERVENTIONS/TREATMENTS ......................................... 45 
5.1 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP)............................................... 45 
5.1.1 
Artemether-lumefantrine (Riamet®) ......................................................... 45 
5.1.2 
Ruxolitinib (Jakavi®) ............................................................................... 45 
5.1.3 
Ruxolitinib Placebo ................................................................................. 46 
5.2 
SELECTION OF DOSE IN THE STUDY .................................................................. 46 
5.3 
SELECTION AND TIMING OF DOSE FOR EACH PARTICIPANT ............................... 46 


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5.4 
DOSE INTERRUPTIONS AND REDUCTIONS.......................................................... 46 
5.5 
SUPPLY, PACKAGING AND LABELLING OF STUDY TREATMENTS ....................... 46 
5.6 
STORAGE OF STUDY TREATMENTS ................................................................... 47 
5.7 
ACCOUNTABILITY, RECONCILIATION AND RETURN OF THE STUDY TREATMENTS
 ..................................................................................................................... 47 
5.8 
PROHIBITED CONCOMITANT THERAPY ............................................................. 47 
5.9 
PERMITTED CONCOMITANT MEDICATION ......................................................... 48 
5.10 
TRIAL INTERVENTION/TREATMENT COMPLIANCE ............................................ 48 
5.11 
MEASURES TO MINIMIZE BIAS .......................................................................... 49 
5.11.1 
Randomisation Procedures ...................................................................... 49 
5.11.2 
Blinding and Unblinding ......................................................................... 49 
6 
STUDY PROCEDURES AND ASSESSMENTS .................................... 49 
6.1 
STUDY CONDUCT SCHEDULE ............................................................................ 49 
6.1.1 
Screening Days -28 to Day -1 .................................................................. 49 
6.1.1.1 
Review of screening/eligibility laboratory testing ........................... 50 
6.1.2 
Eligibility/Safety/Admission Day -1 ......................................................... 50 
6.1.3 
Days 1-4 Treatment with IMP and Confinement ..................................... 51 
6.1.3.1 
Day 1 Pre-first dose IMP ................................................................. 51 
6.1.3.2 
IMP dosing ....................................................................................... 52 
6.1.3.3 
Confinement .................................................................................... 52 
6.1.4 
Discharge to EOS Outpatient Visits ........................................................ 52 
6.1.5 
End of Study Visit (EOS) Day 29 or Early Termination Visit ................. 53 
6.2 
PERMITTED TIME WINDOWS FOR SAMPLING .................................................... 54 
6.3 
PHARMACODYNAMIC ASSESSMENTS ................................................................ 54 
6.3.1 
pSTAT3 .................................................................................................... 54 
6.4 
PHARMACOKINETICS......................................................................................... 54 
7 
SAFETY MONITORING ......................................................................... 55 
7.1 
PHYSICAL EXAMINATION .................................................................................. 55 
7.1.1 
Full Physical Examination ...................................................................... 55 
7.1.2 
Abbreviated Physical Examination .......................................................... 55 
7.1.3 
Symptom Directed Physical Examination................................................ 55 
7.2 
VITAL SIGNS AND OTHER BODY MEASUREMENTS............................................ 56 
7.2.1 
Vital signs ................................................................................................ 56 
7.2.2 
Other Body Measurements – Respiratory rate and Temperature ........... 56 
7.2.3 
Other Body Measurements – Weight and Height .................................... 56 
7.3 
12-LEAD ECGS................................................................................................. 56 
7.4 
CLINICAL LABORATORY TESTS ........................................................................ 57 
7.4.1 
Biochemistry and Haematology Safety Laboratory Testing .................... 57 
7.4.1.1 
Haematology Parameters ................................................................. 57 
7.4.1.2 
Biochemistry Parameters ................................................................. 58 
7.4.1.3 
Coagulation ...................................................................................... 58 
7.4.2 
Urinalysis Safety Laboratory Testing ...................................................... 58 
7.4.3 
Pregnancy and Follicle Stimulating Hormone Testing ........................... 59 
7.4.4 
Serology and Tuberculosis Testing .......................................................... 59 
7.4.5 
Alcohol and Drug Screening ................................................................... 59 
7.5 
BECK DEPRESSION INVENTORY ........................................................................ 60 
7.6 
ADVERSE EVENTS (AES) .................................................................................. 60 
7.6.1 
Definitions ................................................................................................ 60 


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7.6.2 
Causal Relationship to Investigational Medicinal Product..................... 61 
7.6.3 
Severity Grading of Adverse Events ........................................................ 62 
7.6.4 
Documentation of Adverse Events ........................................................... 62 
7.6.5 
Adverse Events of Special Interest ........................................................... 63 
7.7 
SERIOUS ADVERSE EVENTS (SAES).................................................................. 64 
7.7.1 
Definitions of Serious Adverse Events (SAEs) ......................................... 64 
7.7.2 
Reporting for Serious Adverse Events (24 hours) ................................... 64 
7.7.3 
Pregnancy ................................................................................................ 65 
7.8 
SAFETY REVIEW COMMITTEE (SRC) ................................................................ 65 
8 
STATISTICAL ANALYSES .................................................................... 66 
8.1 
GENERAL APPROACH ........................................................................................ 66 
8.2 
SAMPLE SIZE .................................................................................................... 66 
8.3 
ANALYSIS SETS AND SUB-SETS ........................................................................ 67 
8.3.1 
Analysis Sets ............................................................................................ 67 
8.3.1.1 
Full Analysis Set .............................................................................. 67 
8.3.1.2 
Safety Set ......................................................................................... 67 
8.3.1.3 
PK Set 67 
8.3.1.4 
pSTAT3 Set ..................................................................................... 67 
8.3.2 
Sub-Sets .................................................................................................... 67 
8.4 
PARTICIPANT DISPOSITION ............................................................................... 67 
8.5 
DEMOGRAPHICS AND BASELINE CHARACTERISTICS ......................................... 68 
8.5.1 
Demographics .......................................................................................... 68 
8.5.2 
Medical History ....................................................................................... 68 
8.5.3 
Prior Medications .................................................................................... 68 
8.5.4 
Eligibility ................................................................................................. 68 
8.6 
PROTOCOL DEVIATIONS .................................................................................... 68 
8.7 
TREATMENT EXPOSURE .................................................................................... 68 
8.8 
SAFETY (PRIMARY ENDPOINT) ......................................................................... 69 
8.8.1 
Safety Endpoints ...................................................................................... 69 
8.8.2 
Adverse Events ......................................................................................... 69 
8.8.3 
Concomitant Medications ........................................................................ 69 
8.8.4 
Laboratory Findings ................................................................................ 70 
8.8.5 
Physical Examination .............................................................................. 70 
8.8.6 
Vital Signs and other Body Measurements. ............................................. 70 
8.8.7 
12-Lead ECG ........................................................................................... 70 
8.8.8 
Beck Depression Inventory ...................................................................... 71 
8.9 
PHARMACOKINETIC ENDPOINTS ....................................................................... 71 
8.9.1 
Pharmacokinetic parameters ................................................................... 71 
8.10 
PHARMACODYNAMIC ENDPOINTS ..................................................................... 71 
8.10.1 
pSTAT3 levels .......................................................................................... 71 
8.11 
INTERIM ANALYSES .......................................................................................... 71 
9 
STUDY ADMINISTRATION .................................................................. 72 
9.1 
ETHICAL CONSIDERATIONS............................................................................... 72 
9.1.1 
Ethical Principles .................................................................................... 72 
9.1.2 
Informed Consent ..................................................................................... 72 
9.1.3 
Investigator and Human Research Ethics Committee (HREC) 
Responsibilities ................................................................................ 72 
9.2 
PROTOCOL ADHERENCE .................................................................................... 72 


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9.3 
PROTOCOL AMENDMENTS ................................................................................ 72 
9.4 
DATA HANDLING AND RECORD KEEPING ......................................................... 73 
9.5 
QUALITY CONTROL AND QUALITY ASSURANCE ............................................... 73 
9.6 
PUBLICATION POLICY ....................................................................................... 73 
10 
REFERENCES........................................................................................... 74 
11 
APPENDICES ............................................................................................ 77 
APPENDIX 1: CLINICALLY ACCEPTABLE RANGES FOR CLINICALLY IMPORTANT STUDY 
INCLUSION/EXCLUSION LABORATORY TESTS ........................................... 77 
 
 


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List of Tables  
Table 1: Schedule of Activities - Overview..................................................................... 10 
Table 2: Schedule of Activities – Detailed Day 1 to Day 4 Confinement and IMP 
Treatment ......................................................................................................................... 13 
Table 3: Schedule of Activities – Detailed Days 8 to EOS Outpatient Visits ................. 16 
 
List of Figures 
Figure 1: Schema ............................................................................................................... 9 
 
 


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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
AE 
Adverse event 
AESI 
Adverse event of special interest 
AL 
Artemether/lumefantrine (riamet®) 
ALP 
Alkaline phosphatase 
ALT 
Alanine aminotransferase  
AMP 
Amphetamines  
ANC 
Absolute neutrophil count 
APAP 
Acetaminophen/paracetamol 
APTT 
Activated partial thromboplastin time 
AST 
Aspartate aminotransferase 
ATC 
Anatomical therapeutic class 
AUClast 
Area under the plasma concentration-time curve from time zero 
to time of last measurable concentration 
AUC 0- 
Area under the plasma concentration-time curve from time zero 
to infinity 
BAR 
Barbiturates  
BDI 
Beck depression inventory 
β-hCG 
Β-human chorionic gonadatropin 
b.i.d 
Twice-daily (medication) 
BMI 
Body mass index 
BZO 
Benzodiazepines 
C 
Drug concentration in the plasma at any given time 
CI 
Confidence interval 
CKD-EPI 
Chronic Kidney Disease Epidemiology collaboration  
CL/F 
Apparent total clearance of the drug from plasma after oral 
administration 
Cmax 
Maximum (peak) plasma drug concentration 
CMI 
Consumer medication information 
COC 
Cocaine  
CRO 
Clinical research organisation 
CSR 
Clinical study report 
CTCAE 
Common terminology criteria for adverse events 
DBP 
Diastolic blood pressure 
DHA 
Dihydroartemisinin 
ECG 
Electrocardiograph  
ECM 
Experimental cerebral malaria 


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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
eCRF 
Electronic case report form 
eGFR 
Estimated glomerular filtration rate 
ELISA 
Enzyme-linked immunosorbent assay 
EOS 
End of study 
FAS 
Full analysis set 
FBC 
Full blood count 
FDA 
(USA) Food & Drug Administration 
FSH 
Follicle stimulating hormone 
GCP 
Good clinical practice 
GGT 
Gamma-glutamyl transferase 
GMP 
Good manufacturing practice 
HBc Ab 
Hepatitis B core antibodies 
HBs Ag 
Hepatitis B surface antigen 
HDL 
High density lipoprotein 
HIV 
Human immunodeficiency virus 
HR 
Heart rate 
HREC 
Human research ethics committee 
IB 
Investigator’s brochure 
ICH 
International conference on harmonisation  
IFN 
Interferon  
IFNAR 
IFNα receptor 
IL-10 
Interleukin 10  
IMP 
Investigational medicine product 
INR 
International normalised ratio 
IUD 
Intrauterine device 
IV 
Intravenous 
JAKs 
Janus kinase family of tyrosine kinases 
λz 
Terminal disposition rate constant/terminal rate constant 
LAM 
List of Approved Medicines 
LC-MS/MS 
Liquid chromatography-mass spectrometry 
LDL 
Low density lipoprotein 
LFT  
Liver function test 
mAMP 
Methamphetamines 
MCC 
Microcrystalline cellulose 
MCV 
Mean corpuscular volume 
MDMA 
3,4-methylenedioxy-methamphetamine 


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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
MedDRA 
Medical dictionary for regulatory activities 
MTD 
Methadone  
NADH 
Nicotinamide adenine dinucleotide 
OPI 
Opiates  
PBMC 
Peripheral blood mononuclear cell 
PBPK 
Physiologically based pharmacokinetic 
PCP 
Phencyclidine  
PD 
Pharmacodynamics 
PI 
Principal investigator 
PICF 
Participant Information Sheet and Informed Consent Form 
PK 
Pharmacokinetics  
PN 
Preferred name  
pSTAT3 
Phospho-signal transducer and activator of transcription 3 
PT 
Prothrombin time  
qd 
Once a day (medication) 
RBC 
Red blood cell 
Rh(D) 
Rhesus D antigen 
rRNA 
Ribosomal ribonucleic acid 
Rux 
Ruxolitinib (jakavi®) 
SAE 
Serious adverse event 
SAP 
Statistical analysis plan 
SBP 
Systolic blood pressure 
SD 
Standard deviation 
SRC 
Safety Review Committee 
SOA 
Schedule of activities 
SOC 
System organ class 
TB 
Tuberculosis 
TCA 
Tricyclic antidepressants 
TEAE 
Treatment emergent adverse event 
Tfh 
T follicular helper (cell) 
TGA 
(Australian) therapeutic goods administration 
THC 
Tetrahydrocannabinol (cannabis) 
TIBC 
Total iron binding capacity 
t½ 
Elimination half-life 
tlag 
Lag time 
tmax 
Time to reach maximum (peak) plasma concentration following 
drug administration 


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LIST OF ABBREVIATIONS AND DEFINITION OF TERMS 
ULN 
Upper limit of normal 
Vz/F 
Apparent volume of distribution of drug during terminal phase 
after non-intravenous administration 
WBC 
White blood cell 
WHO 
World health organization 
WOCBP 
Women of childbearing potential 
 
 


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PROTOCOL APPROVAL/ SIGNATURES 
I herewith approve the following protocol entitled “A randomised, single blind, placebo 
controlled, phase 1 trial to evaluate the safety, tolerability, pharmacokinetic and 
pharmacodynamic activity of Ruxolitinib when co-administered with artemether-
lumefantrine in healthy participants”, Version 4.0 dated 07 September 2020. 
 
SPONSOR SIGNATURE 
 
 Signature: 
_______________________________________________ 
Name: 
Dr Farouk Chughlay MD 
Role: 
Medical and Project Director, Medicines for Malaria 
Venture 
Date: 
____/____/2020 
 
DOCUMENT HISTORY 
Document 
History 
Amendment 3 
V4.0 07 September 2020 
Amendment 2 
V3.0 07 August 2020 
Amendment 1 
V2.0 16 July 2020 
Original Protocol 
V1.0 18 June 2020 
 
 


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PRINCIPAL INVESTIGATOR SIGNATURE 
 
A randomised, single blind, placebo controlled, phase 1 trial to evaluate the safety, 
tolerability, pharmacokinetic and pharmacodynamic activity of Ruxolitinib when 
co-administered with artemether-lumefantrine in healthy participants  
MMV_Ruxolitinib_19_01 
 
Version: 4.0 
 
Issue Date: 07 September 2020 
 
Principal Investigator Agreement 
 
I have read the above-mentioned protocol and am aware of my responsibilities as Principal 
Investigator for this study. As such, I agree to: 
• Personally supervise the conduct of this trial; 
• Conduct the trial in accordance with International Conference on Harmonization 
(ICH) E6 Good Clinical Practice: Consolidated Guidance (GCP), applicable 
regulatory requirements, and the protocol; 
• Comply with the procedures for data recording and reporting as required by the 
regulatory authorities and the Sponsor; 
• Permit monitoring, auditing, and inspection of study records as required by ICH 
GCP; 
• Retain the essential clinical study documents as required by ICH GCP and the 
Sponsor. 
 
Principal Investigator Signature: 
 
Principal Investigator Name: 
Dr Paul Griffin 
Date: 
____/____/2020 
 
 


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2 INTRODUCTION AND RATIONALE 
2.1 
DISEASE BACKGROUND 
Malaria is a mosquito-borne parasitic infection that is endemic in tropical and sub-tropical 
countries globally.1,2 The infective organism is the protozoan Plasmodia species, with 5 
species known to cause human disease (P. falciparum, P. vivax, P. ovale, P. malariae and 
P. knowlesi).1,3 P. falciparum is the significant sub-species causing the majority of 
mortality and morbidity globally.1 The life cycle of the malaria parasite consists of a liver 
stage and a blood stage within the human host, the latter being the cause of the clinical 
symptoms experienced in malaria infection.4 Malaria is still a major global health issue, 
and remains a significant health problem in tropical and sub-tropical regions of the world. 
As such, the World Health Organisation (WHO) has declared malaria a global health 
priority.1 According to the WHO 2019 Annual Malaria Report, there were an estimated 
228 million cases of malaria globally in 2018 which resulted in 405 000 deaths, and 
malaria remains responsible for more deaths than any other parasitic disease.1  
Substantial progress in disease control has been made through vector management, use of 
insecticide-treated bed nets, and improved diagnosis and drug treatment.1  Critically, there 
is no licensed vaccine, and the emergence of artemisinin-resistant parasites in the Greater 
Mekong Subregion represents a serious threat to malaria treatment and control efforts. A 
major impediment to malaria eradication currently is poor understanding of host 
immunity against the Plasmodium species. There is a lack of consensus on how to 
generate long-lasting immunity in malaria endemic areas, and poor performances to date 
of malaria vaccine candidates in such endemic settings.  
Studies performed more than 50 years ago showed anti-parasitic antibodies can protect 
against clinical malaria and control parasite growth.2 However, these responses take years 
to develop in malaria endemic areas, after repeated exposure to malaria parasites.3 Recent 
reports also highlight a requirement for antibodies with specific functional properties to 
mediate immunity.4 The development of long-lived, anti-parasitic memory B and plasma 
cells that produce these antibodies requires CD4+ T cell help.5,6 Specialised CD4+ T 
follicular helper (Tfh) cells are critical in this process by providing key cognate and 
soluble signals to B cells.7,8 Interferon gamma (IFNγ) production by Tbet+ CD4+ T (Th1) 
cells is also important for generating specific antibody isotypes and activating phagocytic 
cells for increased antibody-mediated and compliment-mediated removal of parasitised 
red blood cells, and generating microbiocidal molecules such as reactive oxygen and 
nitrogen intermediates.9,10  
However, these anti-parasitic responses are often absent, sub-optimal, or dysfunctional in 
individuals living in malaria endemic areas.11-13 The reasons for this are still unclear, but 
emerging evidence indicates parasite-induced development of atypical B cells, sub-
optimal Tfh cells, and Th1 cell responses characterised by autologous interleukin 10 (IL-
10) production (type 1 regulatory [Tr1] cell responses), as well as the development of 
immunoregulatory networks, all contribute to these outcomes.14-19  
Recently, type I interferons (IFNs) have emerged as important regulators of IL-10 
production by Type 1 regulatory T (Tr1) cells. Type I IFNs comprise a large family of 
cytokines that signal through the common IFNα receptor (IFNAR), consisting of IFNAR1 
and IFNAR2 chains. The IFNAR signals via signal transducers and activators of 
transcription 1 and 2 (STAT-1 and STAT-2) to mediate diverse functions during many 
infections.20-22 Polymorphisms in the IFNAR1 gene have been associated with increased 


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risk of severe malaria in The Gambia, while nonclinical murine studies identified a type I 
IFN-dependent transcriptional program associated with the pathogenesis of severe 
malaria experimental cerebral malaria (ECM).23-25 Type I IFNs suppress CD4+ T cell-
dependent parasite control during experimental blood stage malaria by modulating the 
function of dendritic cells following P. berghei ANKA infection, rather than acting 
directly on Th1 cells.26,27 Another trial in mice infected with P. yoelli showed that type I 
IFNs directly promoted the expansion of Tr1 cells.28  
Significantly, in malaria volunteer infection studies (VIS), type I IFNs produced by 
several different cell sources were found to be important regulators of developing anti-
parasitic immunity. Type I IFNs not only suppressed innate immune cell function and 
parasitic-specific CD4+ T cell IFNγ production, but also promoted the development of 
parasitic-specific Tr1 cells.29 Thus, type I IFNs are key immunomodulatory molecules in 
humans infected with P. falciparum, and targeting this pathway for clinical advantage is 
a promising strategy to overcome established or developing immunoregulatory networks 
to improve natural, drug-mediated, or vaccine-induced immunity against malaria.  
Ruxolitinib (Jakavi®) blocks type 1 IFN signaling, and is therefore a candidate for 
investigation in combination with established anti-malarial medications such as Riamet®. 
If the results of this study provide an acceptable safety, tolerability, pharmacokinetic and 
pharmacodynamic profile for the combination of ruxolitinib and Riamet®, a future malaria 
Volunteer Infection Study (VIS) / Controlled Human Malaria Infection (CHMI) study is 
planned to evaluate the combination’s immune enhancing potential in malaria.   
Eradication of malaria will not be possible until continuing research in these areas can 
provide further information. 
2.2 
INVESTIGATIONAL MEDICINAL PRODUCTS  
2.2.1 Ruxolitinib (Jakavi®) 
2.2.1.1 
Clinical Studies 
Ruxolitinib (Rux) is a licensed orally administered small molecule inhibitor of Janus 
kinase JAK1/2, and has an established safety profile that includes Phase 1 study 
participants with a relatively short elimination half-life (~3 hours). Rux is registered for 
the treatment of intermediate or high-risk myelofibrosis in adults, and has also been safely 
and effectively used in children with type I interferonopathy.30  
The Janus kinase family of tyrosine kinases (JAKs) are intimately associated with 
cytokine receptors such as the type I IFN receptor. The JAKs become phosphorylated 
after cytokines bind to these receptors, and in turn create binding sites for STATs. The 
STATs are then phosphorylated by JAKs, dissociate from the cytokine receptor and 
translocate to the cell nucleus to drive transcription of target genes.31 Following the 
binding of IL-6 to the IL-6 receptor, JAK1 and JAK2 are phosphorylated leading to 
phosphorylation of STAT3 (pSTAT3).  This response in monocytes and T cells can be 
used to measure the pharmacodynamic (PD) effect of Rux.32  
Rux is used to treat neoplastic diseases, but has also been used to successfully treat 
children with a type I interferonopathy.33-35 In addition, Rux was found to reduce serum 
type I IFN levels and IFN-inducible gene scores in dermatomyositis patients.30 Thus, there 
is a growing body of evidence that Rux may be used to block type I IFN signaling in 
humans in a range of diseases, including malaria. 30,35 


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Rux is also suitable for application in malaria endemic settings either in combination with 
anti-parasitic drugs (such as Riamet®) and/or malaria vaccines. The short half-life of Rux 
will minimise risk of adverse outcomes in populations where co-infections with 
tuberculosis, human immunodeficiency virus (HIV) and other pathogens is relatively 
common. 
2.2.2 Riamet® - Artemether/lumefantrine (AL)   
Riamet® is a registered, standard drug widely used for the treatment of malaria, and is a 
fixed combination of artemether and lumefantrine (AL). Artemether inhibits an essential 
calcium adenosine triphosphatase, while the exact mechanism of lumefantrine is 
unknown.   
Any impact of AL on Rux activity will be assessed by measuring pSTAT3 inhibition in 
blood samples taken from participants, as previously described in phase 1 safety studies.32  
2.2.3 Clinical Safety and Risk Assessment  
Individual risks pertaining to Riamet® and Jakavi® have been well researched. The 
Sponsor conducted physiologically based pharmacokinetic (PBPK) modelling 
simulations, and concluded that the risk for a clinically relevant drug-drug interaction 
between the two drugs was remote especially when administering Rux 2 hours after AL.36 
Toxicity rules for IMPs and other trial interventions have been specifically developed to 
manage the risk of their use in this study (see Section 4.3.8 below). Risk will be minimised 
through comprehensive screening of participants against robust inclusion and exclusion 
criteria encompassing screening for infectious diseases including tuberculosis. 
Participants will be monitored closely throughout the study with clinical laboratory tests, 
physical examinations, vital signs, and ECG, including a period of confinement at the 
clinical unit at the time of IMP administration.  Adverse Events (AEs) will be recorded 
throughout the study and the safety of the study will be reviewed by the Principal 
Investigator, MMV Medical Director and study Medical Monitor after the sentinel group 
(Group 1a) has completed Day 8. 
2.2.3.1 
Ruxolitinib  
Review of healthy volunteer data emphasised that possible haematological effects 
including reduction in absolute neutrophil count (ANC) are monitorable, low grade, 
predictable in terms of timing and grade, directly linked to the effect of the drug. In 
particular, at the proposed regimen, a decrease in ANC is expected to be transient and due 
to margination rather than a direct toxic effect.  
The safety, tolerability, and pharmacokinetics (PK) of Rux has been studied in healthy 
volunteers including males and females (including women of child-bearing potential 
[WOCBP]).32 The PK, PD, safety, and tolerability of orally administered Rux has been 
evaluated in 2 double-blind, randomised, and placebo-controlled healthy volunteer studies 
conducted in the United States of America. The first study enrolled 23 participants and 
evaluated single ascending doses of 5 to 200 mg and the effect of food. The most frequent 
AEs reported were headache (13.0% overall); diarrhoea (13.0% overall), and blood 
sampling catheter site haemorrhage (13.0% overall). There was no dose dependency for 
the frequency of AEs as a whole or of any particular AE. 
The second study enrolled 71 participants and evaluated multiple ascending doses, 
including both once- and twice-daily dosing for 10 days. Neutropenia of any severity 
grade was observed in 11.1% of participants at 50 mg qd, 66.6% of participants at 100 mg 


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qd, 12.5% of participants at 15 mg b.i.d, 33.3% of participants at 25 mg bid, 66.6% of 
participants at 50 mg b.i.d and 22.2% of placebo participants. These were generally 
transient and rapidly normalised following the last dose of study medication. One 
participant who received Rux 50 mg bid discontinued the study due to grade 4 
neutropenia. In both these single- and multiple-dose studies, AEs were generally mild to 
moderate in intensity and resolved quickly. The 1 episode of grade 4 neutropenia in the 
participant who received 50 mg bid was considered a dose-limiting toxicity. 
The safety, tolerability, and PK of Rux was further evaluated in healthy Japanese 
volunteers.37 Forty study participants were randomised to receive single (10–100 mg) and 
multiple (10 and 25 mg every 12 h) doses of Rux or placebo. AEs occurred in 15% of the 
participants overall; were either grade 1 or 2 in severity and were considered related to 
study drug. The majority of AEs were observed in participants during treatment with 
repeated b.i.d doses, during which the observation period was longer, but there was no 
clear pattern of dose dependency. Grade 2 neutropenia occurred in five (12.5%) Rux 
treated participants, however decreased neutrophils were also observed in all placebo 
treated participants. Neutrophil counts returned to normal within 24 h of the last dose for 
all participants. No dose discontinuations or SAEs occurred, however dose adjustments 
were made in two Rux b.i.d treated participants due to decreased neutrophil counts on day 
5. 
In summary, orally administered Rux was considered safe and well tolerated in healthy 
volunteers, with 25 mg b.i.d and 100 mg qd established as the maximum tolerated doses. 
There were no apparent differences in the safety or PK of Rux between Japanese and non-
Japanese study participants. 
Rux has no known contraindications. Detailed information about Jakavi® is available in 
the Product Information and the Consumer Medical Information (CMI) sheet.   
2.2.3.2 
Riamet® - Artemether/lumefantrine (AL) 
In this study, the dosing regimen for AL will be as recommended for an adult treated for 
acute uncomplicated malaria in the Product Information and/or CMI.  Detailed 
information about the risks of using Riamet® is also available in the Product Information 
and/or CMI sheet.  
Briefly, adverse effects that have been reported from using AL include: stomach pain, 
diarrhoea, dizziness, aching muscles/joints, sore throat/cough, nausea/vomiting, 
headache, difficulty sleeping, tingling, and fever/shivering. Most of these side effects 
have been mild and transient.   
2.3 
RATIONALE 
2.3.1 Rationale for the Study 
The World Health Organization (WHO) estimated 228 million cases of malaria and 
405,000 deaths from malaria occurred worldwide in 2018. The WHO African Region 
carries the highest burden, with 213 million cases (93%) accounting for 94% of all malaria 
deaths in 2018. Nearly all of the malaria cases (99.7%) in sub-Saharan Africa were caused 
by the parasite Plasmodium falciparum.1 Current approaches to malaria control are 
failing, and emerging evidence suggests that both developing and established 
immunoregulatory pathways impede the acquisition of natural, drug-mediated, or 
vaccine-induced immunity against malaria. Type I interferons have been identified as 
important immune regulators in malaria, and are promising targets for improving anti-
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Ruxolitinib (Jakavi®) has shown immune-enhancing ability when combined with a drug 
targeted at the pathogen of interest. The purpose of this Phase 1 study is to investigate if 
co-administration of ruxolitinib and the antimalarial drug Riamet® (artemether-
lumefantrine; AL) can be safely conducted in a larger proof of concept trial against P. 
falciparum malaria, and is safe, tolerable, and has any effect on the pharmacodynamics 
of ruxolitinib.  
The study is designed to assess the safety, tolerability, pharmacokinetics (PK) and 
pharmacodynamics (PD) (pSTAT3) of oral doses of artemether-lumefantrine (AL; 20 
mg/120 mg) and Ruxolitinib (Rux; 20 mg), or oral doses of artemether-lumefantrine (AL; 
20 mg/120 mg) and placebo. Ruxolitinib or placebo will be administered 2 h after AL 
(i.e., AL+Rux or AL+placebo) twice daily (b.i.d) for 3 consecutive days in healthy adult 
volunteers.  
2.3.2 Rationale for the Dose 
The dose of 20 mg Rux b.i.d. to be used in the study is the standard dose for patients with 
platelet count >200 x 109/L in the Australian Product Information (PI) for Jakavi®. A 3-
day 20 mg b.i.d. dosing regimen was considered appropriate for healthy volunteers based 
on the reported safety of a higher dose of 25 mg b.i.d. over a 10-day period in healthy 
volunteers in a phase 1 safety trial.32 No effect of food was found in this trial, and 
consequently Rux/placebo can be administered without regard to meals.  
The co-administration of AL and Rux was chosen to mimic how Rux is likely to be used 
in field settings with patients receiving treatment for an episode of acute uncomplicated 
malaria. The 3-day dosing of Rux in combination with AL was based on an 
immunological rationale for combining the therapies, involving AL-related killing of 
parasites leading to activation of type 1 IFN pathways to be disrupted by Rux.  
The dose regimen for AL to be used in the trial is the standard adult dose as described in 
the Product Information and/or Consumer Medication Information (CMI) sheet.  
 
 


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3 STUDY OBJECTIVES AND ENDPOINTS 
 
Primary objective  
Primary endpoint 
1. To assess the safety and 
tolerability of 3-day b.i.d dosing of 
AL+Rux and AL+placebo.  
1. Incidence, severity, and relationship of 
observed and self-reported AEs up to 28 days 
after 
AL+Rux 
and 
AL+placebo 
administration in all participants by treatment 
regimen.  
2. Clinical laboratory evaluations including 
haematology, biochemistry, and urinalysis. 
3. Vital signs (blood pressure and heart rate) 
4. 12-lead standard ECG: QT, QTcB and QTcF; 
HR, PR, QRS. 
Secondary objectives 
Secondary endpoints 
1. To assess the effect of AL+Rux 
and 
AL+placebo 
on 
pSTAT3 
inhibition.  
1. pSTAT3 inhibition ex-vivo on whole blood 
cells by: 
• pSTAT3 levels pre- and post- AL+Rux 
administration intra participant and, 
• pSTAT3 levels per treatment regimen post 
AL+Rux 
and 
post 
AL+placebo 
administration  
2. To characterise the PK profile of 
artemether and its major metabolite 
dihydroartemisinin 
[DHA], 
lumefantrine, and Rux (part 1 and 2). 
2. PK parameters of artemether, DHA, 
lumefantrine, 
and 
Rux 
using 
non-
compartmental methods: AUClast, AUC0-, 
AUC0-8, AUC60-72, Cmax (first and last dose), tmax 
(first and last dose), elimination half-life (t½), 
tlag, C168h (for lumefantrine only), CL/F, Vz/F 
and λz in all participants. 
3. To assess additional safety 
variables 
(tympanic 
body 
temperature and respiration rate) 
3. Safety variables tympanic body temperature 
(°Celcius) and respiratory rate. 
 
 


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4 INVESTIGATIONAL PLAN 
4.1 
DESCRIPTION OF OVERALL STUDY DESIGN AND PLAN 
This study is a randomised, single-blind, placebo-controlled, single centre phase 1 trial 
designed to assess the safety, tolerability, pharmacokinetics (PK) and pharmacodynamics 
(PD) of combined oral doses of:  
• artemether-lumefantrine (AL; 20 mg/120 mg) and Ruxolitinib (Rux; 20 mg), or  
• artemether-lumefantrine (AL; 20 mg/120 mg) and placebo. 
The study will be composed of 2 groups (Group 1a and Group 1b) to be enrolled 
sequentially. Group 1a will participate first as the sentinel safety group.   
4.1.1 Group 1a (sentinel group) 
Group 1a will be composed of 2 participants.  
After informed consent, screening and confirmation of eligibility: 
• Participants will be admitted to the clinical trial unit on Day -1.  
• One participant will be randomised to receive AL+Rux and the other participant will 
be randomised to receive AL+placebo. Randomisation will be single blinded 
(treatment allocation will be concealed to the participants but not to the Investigator). 
• On Day 1, participants will be administered oral doses of AL, with oral doses of 
Ruxolitinib or placebo administered 2 hours later twice daily (b.i.d) for 3 consecutive 
days (AL at t = 0, 8, 24, 36, 48 and 60 hours, Rux at t = 2, 10, 26, 38, 50 and 62 
hours). 
• Participants will be confined to the clinical trial unit for approximately 4 days 
(4 nights) from admission on Day -1 for blood sampling for PK and PD (pSTAT3) 
and safety monitoring.  
• After discharge from the clinical trial unit no less than 78 hours post-first dose IMP, 
participants will be followed up via outpatient monitoring visits for safety 
assessments, pSTAT3 blood sampling, and PK blood sampling up to the end of study 
visit (EOS) on Day 29.  
• The decision to proceed with Group 1b will be made by the Principal Investigator, 
Sponsor Medical Director, and the study Medical Monitor (at a minimum) after 
review of Group 1a’s safety and tolerability data up to and including Day 8.  
4.1.2 Group 1b 
If the Principal Investigator, Sponsor Medical Director and the study Medical Monitor (at 
a minimum) confirm Group 1b will proceed, it will be composed of 6 participants: 
• 5 participants will be randomized to AL+Rux and 1 participant will be randomized 
to AL+placebo. Randomisation will be single blinded (treatment allocation will be 
concealed to the participants but not to the Investigator). 
• Study participation will otherwise be as described in Section 4.1.1. 
4.2 
DISCUSSION OF STUDY DESIGN INCLUDING CHOICE OF CONTROL 
GROUPS 
The study is designed specifically to monitor safety and tolerability of the combination of 
AL+Rux, and to obtain PK and preliminary PD data. Eight participants will be 
randomised to receive AL+Rux or AL+placebo in a 3:1 ratio overall (1:1 in Group 1a, 


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and 5:1 in Group 1b). This is typical of first-in-human studies, and this approach is used 
in this study as the combination of AL+Rux has not been studied previously.  
4.3 
SELECTION OF STUDY POPULATION 
Individuals must fulfil all of the following inclusion criteria and none of the exclusion 
criteria to be eligible for inclusion in this trial. No exemptions or protocol waivers will be 
granted. 
4.3.1 Inclusion Criteria 
 
INCLUSION CRITERIA 
Demography 
1. Male or female (non-pregnant, non-lactating) aged 18 to 55 years inclusive.  
2. Contactable and available for the duration of the trial and for up to two weeks 
following the EOS visit. 
3. 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) at Screening and Day -1. 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 
4. Certified as healthy by a comprehensive clinical assessment (detailed medical 
history, full physical examination and special investigations). 
5. Vital signs 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. ECG parameters for both males and females: QT ≤ 500 msec, QTcF ≤450 msec, 
QTcB ≤450 msec; PR interval ≤210 msec. 
7. Heterosexual female participants of childbearing potential who have, or may have, 
male sexual partners during the course of the study should be using an insertable 
(implant or IUD), injectable, transdermal or combination oral contraceptive 
approved by the TGA combined with a barrier contraceptive from the time of 
informed consent until 30 days after EOS. Abstinent female participants must agree 
to start a double method if they start a sexual relationship with a male during the 
trial until 30 days after EOS. Female participants must not be planning in vitro 
fertilisation within the required contraception period. 
Women of non-childbearing potential who will not require contraception during the 
trial are defined as: surgically sterile (tubal ligation is not considered surgically 
sterile), post-menopausal (spontaneous amenorrhoea for ≥12 months, or 
spontaneous amenorrhoea for 6-12 months and follicle-stimulating hormone (FSH) 
≥40 IU/mL; either should be together with the absence of oral contraceptive use for 
>12 months). 
Male participants who have, or may have female sexual partners during the course 
of the study must agree to use a double method of contraception including condom 
plus diaphragm, or condom plus stable insertable (implant or IUD), injectable, 
transdermal or combination oral contraceptive by the female partner, from the time 
of informed consent through to 90 days after EOS. Abstinent male participants must 


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INCLUSION CRITERIA 
agree to start a double method if they begin a sexual relationship with a female 
during the trial, and through to 90 days after EOS. Male participants with female 
partners that are surgically sterile or post-menopausal, or male participants 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. 
In addition, all male participants must agree to not donate sperm for at least 90 days 
after EOS.  
Regulations 
8. Completion of the written informed consent process prior to undertaking any trial-
related procedure.  
9. Must be willing and able to communicate and participate in the whole trial. 
10. Agree to adhere to Lifestyle Considerations (see Section 4.3.3) throughout trial 
duration and be willing to consume 250 mL full-fat milk with each dose of AL. 
4.3.2 Exclusion Criteria 
If any of the following exclusion criteria apply, the individual will not be permitted to 
participate or remain in the study.  
EXCLUSION CRITERIA 
Medical history and clinical status 
1. Known hypersensitivity to ruxolitinib, artesunate or any of its excipients, 
artemether, lumefantrine or other artemisinin derivatives, proguanil/atovaquone, 
primaquine, or 4-aminoquinolines. 
2. Haematology, biochemistry or urinalysis results that are abnormal/outside of the 
laboratory normal reference ranges AND are either: 
• considered clinically significant by the Principal Investigator or delegate; OR 
• considered not clinically significant by the Principal Investigator or delegate 
BUT ARE ALSO outside of Sponsor-approved clinically acceptable laboratory 
ranges in Appendix 1.  
NOTE: Participants are not excluded if abnormal/out of laboratory normal reference 
range results are considered not clinically significant by the Principal Investigator 
or delegate AND are within the ranges specified in Appendix 1.  
3. Platelets < 200 x109/L at Screening or prior to IMP administration is exclusionary. 
One re-test is permitted if original test results does not reflect the assumed medical 
status of the individual.  
4. Participation in any other investigational product trial within 5 half-lives or the 12 
weeks preceding IMP administration, whichever is longer, or in the exclusion period 
of a previous trial according to applicable regulations.  
5. Symptomatic postural hypotension at screening and pre-first dose of IMP on Day 1 
(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. 


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EXCLUSION CRITERIA 
6. History or presence of diagnosed (by an allergist/immunologist) or treated (by a 
physician) food or known drug allergies, or any history of anaphylaxis or other 
severe allergic reactions including face, mouth, or throat swelling or any difficulty 
breathing. 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 is not an exclusion criterion. 
8. Presence of current or suspected serious chronic diseases such as cardiac or 
autoimmune disease (HIV or other immuno-deficiencies), insulin-dependent and 
non-insulin-dependent 
diabetes, 
progressive 
neurological 
disease, 
severe 
malnutrition, acute or progressive hepatic disease, acute or progressive renal 
disease, porphyria, psoriasis, rheumatoid arthritis, asthma (excluding childhood 
asthma), epilepsy, or obsessive-compulsive disorder. 
9. History of malignancy of any organ system (other than localised and considered 
cured basal cell carcinoma of the skin or in situ cervical cancer), treated or untreated, 
within five years of screening, regardless of whether there is no 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. 
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 (BDI-II) will be used as a validated tool for the 
assessment of depression at screening. In addition to the conditions listed above, 
individuals 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 individuals will be referred to a general practitioner or medical 
specialist as appropriate. Individuals 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 individual 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. Acute illness within the 4 weeks prior to screening and prior to IMP administration.  
15. Significant inter-current disease of any type, in particular liver, renal, cardiac, 
pulmonary, neurologic, rheumatologic, or autoimmune disease by history, physical 
examination, and/or laboratory studies including urinalysis. 


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EXCLUSION CRITERIA 
16. Individual has a clinically significant disease or any condition or disease that might 
affect drug absorption, distribution or excretion (e.g. gastrectomy, cholecystectomy, 
diarrhoea) or known lactose/dairy intolerance. 
17. Participation in any research trial involving blood sampling (more than 300 mL/unit 
of blood) within one month prior to IMP administration, or blood donation to 
Australian Red Cross Blood Service (Blood Service) or other blood bank during the 
8 weeks prior to IMP administration. 
18. Medical requirement for intravenous immunoglobulin or blood transfusions. 
19. 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. 
20. Any individual who has ever smoked >1 pack of cigarettes per day for >10 years, 
or who currently (within 14 days prior to Screening or prior to IMP administration 
smokes >5 cigarettes/day. 
21. Female who is breastfeeding.   
22. Any vaccination within the last 28 days prior to screening or prior to IMP 
administration. 
23. Prior to screening or IMP administration: any systemic or inhaled 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 topical, 
nasal or ophthalmic corticosteroids within the past 2 weeks. 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. 
24. Use of antidepressant medication in the past 12 months prior to screening or prior 
to IMP administration. 
25. Use of any other medication except contraceptives (including herbal, vitamin 
supplement, OTC or prescription) within 14 days or five half-lives (whichever is 
longest) prior to IMP administration. Participants are requested to refrain from 
taking non-approved concomitant medications from recruitment until the 
conclusion of the trial. 
26. Cardiac/QT risk: 
• Family history of sudden death or of congenital prolongation of the QT, QTcF, 
or QTcB interval or known congenital prolongation of the QT, QTcF, or QTcB 
interval or any clinical condition known to prolong the QT, QTcF, or QTcB 
interval. 
• History of symptomatic cardiac arrhythmias or with clinically relevant 
bradycardia. 
• Electrolyte disturbances, particularly hypokalaemia, hypocalcaemia, or 
hypomagnesaemia. 
• ECG abnormalities in the standard 12-lead ECG (at screening or prior to IMP 
administration) which in the opinion of an Investigator is clinically relevant or 
will interfere with the ECG analyses.  


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EXCLUSION CRITERIA 
General conditions 
27. 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. 
28. Any individual for whom study participation would pose an additional safety risk 
as assessed by the Principal Investigator.  
29. Any individual who is an Investigator, research assistant, pharmacist, trial 
coordinator, or other staff thereof, directly involved in conducting the trial. 
30. Any individual without good peripheral venous access. 
Biological status 
31. 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). 
32. Recent herpes zoster infection (within the previous 6 months) as determined by 
clinical history. 
33. Positive result for M. tuberculosis infection by QuantiFERON-TB Gold assay. 
34. Positive urine drug test for any drug listed in Section 7.4.5. Any individual testing 
positive for acetaminophen (paracetamol) at screening and/or pre-dose  may still be 
eligible for trial participation at the discretion of the Principal Investigator or 
delegate. 
35. Positive alcohol breath test. 
 
 


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4.3.3 Lifestyle Considerations 
While participating in this trial, participants are asked to: 
• Refrain from alcohol consumption of more than 40 g/4 units/4 standard drinks per 
day, and/or drug habituation until the conclusion of the trial, and also: 
• Abstain from any alcohol use for the duration of the clinical unit confinement.  
• Refrain from tobacco use of more than five cigarettes or equivalent per day until the 
conclusion of the trial, and also: 
• Abstain from any tobacco use for the duration of clinical trial unit confinement. 
• Refrain from excessive consumption of beverages or food containing xanthine bases 
including Red Bull, chocolate, coffee, black tea etc. (more than 400 mg caffeine per 
day, equivalent to more than 4 cups of coffee per day), and also: 
• Abstain from any consumption of beverages or food containing xanthine bases for 
the duration of clinical trial unit confinement. 
• Refrain from consumption of Seville oranges, and grapefruit or grapefruit juice from 
7 days prior to IMP dose until the EOS.  
• Refrain from consuming poppy seeds one week prior to Screening and for the 
duration of the study.  
• Abstain from strenuous exercise for 24 h before each blood collection for clinical 
laboratory tests. Participants may participate in light recreational activities during 
studies. 
Clinical study staff will routinely remind participants of these requirements.   
4.3.4 Screen Failures 
Healthy candidate volunteers who do not fulfil all the inclusion criteria, and/or fulfil any 
of the exclusion criteria should not be enrolled into the trial without exception. In case of 
doubt, the Principal Investigator or delegate is to confer with the Medical Monitor for 
agreement. Waivers for inclusion of volunteers who are not meeting all eligibility criteria 
will not be granted. If a participant does not meet all selection criteria (is a screen failure) 
but at some point in the future is expected to meet the eligibility criteria, the participant 
may be rescreened on 1 occasion only. Participants who are rescreened will undergo the 
informed consent process, be assigned a new participant number, and then restart a new 
screening phase. 
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. 
Re-tests are limited to one per parameter (except for urine drug screen), and only if the 
Principal Investigator considers that the existing test result may not correctly reflect the 
participant’s medical status. The result of the retest must be considered for participant 
eligibility and must be available prior to Day 1. Reasons for re-testing must be clearly 
documented. A positive urine drug screen is exclusionary (except if positive for 
paracetamol only at the discretion of the Principal Investigator or delegate) and no re-
testing is permitted. Findings made during unscheduled visits will be documented. 
Minimum information to be collected on screen failures include demography, details of 
the screen failure, eligibility criteria and any SAE, and will NOT be entered into the eCRF.   
Participants who fail screening due to an underlying medical condition (acute self-limiting 
viral infections do not apply) previously unknown to them will be reimbursed for their 


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time, and provided with the appropriate referrals for guidance and counselling for their 
condition. 
4.3.5 Withdrawal of Participants 
Participants are free to withdraw from participation in the trial at any time upon request.  
The Sponsor and/or Principal Investigator or delegate may discontinue or withdraw a 
participant from the trial for the following reasons: 
• Pregnancy (participant must be withdrawn from the study) 
• Significant trial intervention non-compliance or protocol deviation (to be decided 
on a case-by-case basis) 
• If any clinical AE, laboratory abnormality, or other medical condition or situation 
occurs such that continued participation in the trial would not be in the best interest 
of the participant (see also Section 4.3.8 regarding toxicity rules) 
• If the participant meets an exclusion criterion (either newly developed or was not 
previously recognised) that precludes further trial participation 
• Participation in any investigational product trial whilst enrolled in this trial (duration 
up to 12 months)  
• If participant was unblinded (on a case by case basis) 
The reason for participant discontinuation or withdrawal from the trial will be recorded 
on the electronic case report form (eCRF) and withdrawn participants will be asked to 
complete an early termination visit (ET). ET procedures are the same as for EOS (see 
Section 6.1.5). 
If a participant withdraws or is withdrawn from the trial, the Principal Investigator or 
delegate will inform the Sponsor immediately. If there is a medical reason for withdrawal, 
the participant will remain under the supervision of the Principal Investigator or delegate 
until satisfactory health has returned, or medical/clinical care has been transferred to the 
participant’s general practitioner or appropriate hospital consultant. 
The Principal Investigator or delegate 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.  
4.3.5.1 
Replacement of Withdrawn/Discontinued Participants 
• Participants who sign the informed consent form and are randomised but do not 
receive IMP may be replaced. Replacement volunteers are permitted as back-up.  
• Participants who sign the informed consent form, and are randomised and receive at 
least one dose of IMP, and subsequently withdraw, or are withdrawn or discontinued 
from the trial, may only be replaced after mutual agreement between the Sponsor and 
the Principal Investigator or delegate. The decision regarding the replacement of 
participants will be documented. 


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4.3.6 Lost to Follow-Up 
For participants who are lost to follow-up, the Principal Investigator or delegate will 
demonstrate due diligence by documenting all steps taken to contact the participant (e.g., 
dates of phone contacts, registered letter, home visit, etc.) in the source documents. 
It will be explained to the participants during the consenting process and throughout the 
duration of the study that they must be readily contactable to the study and clinical trial 
unit team.   
4.3.7 Study Discontinuation 
The Sponsor, Principal Investigator, SRC, approving HREC, and regulatory authorities 
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 (the Sponsor will notify regulatory authorities as appropriate). After a decision 
to prematurely terminate the trial or to suspend the trial, the Sponsor and the Principal 
Investigator will ensure that adequate consideration is given to protecting the participants’ 
interest and safety. The Principal Investigator or delegate must review all participants as 
soon as practicable and complete all required records.  
4.3.8 Trial Intervention/Treatment Discontinuation – Toxicity Rules 
In addition to the classic assessment of SAEs (see Section 7.7) and consideration of 
adverse events of special interest (AESIs – see Section 7.6.5), specific toxicity rules apply 
for this trial and must be applied when early termination or suspension is considered.  
Toxicity rules 
When applying toxicity rules, AEs will be graded using the Common Terminology 
Criteria for Adverse Events (CTCAE) grading system (Version 5.0 Published: November 
27, 2017) for standardised recording and reporting. For the purpose of this protocol, 
“toxicity” refers to the occurrence of any grade 3 or higher grade AE that is related to an 
IMP (AL, Rux, and Placebo).  
General toxicity rules 
General toxicity rules refer to all AEs, except for AEs relating to the liver and 
haematology. General toxicity rules also apply to neutropenia, as it is an identified liability 
of Rux.  
Trial-specific toxicity rules applying to AEs relating to the liver and haematology have 
also been created for this trial, as the CTCAE grading system may not give clear enough 
guidance.  
4.3.8.1 
Individual Toxicity Rules 
IMP administration for an individual participant will be stopped if: 
• the participant experiences any CTCAE grade 3 (or higher) AE, or an SAE 
irrespective of severity/CTCAE grade deemed related to the IMP. 
• the participant experiences any CTCAE grade 1 or 2 AE that raises a safety 
concern and is deemed related to IMP. 


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4.3.8.2 
Group Toxicity Rules 
IMP administration for a group will be stopped if the following occurs in more than one 
participant: 
• any CTCAE grade 3 (or higher) AE, or an SAE deemed related to the IMP 
irrespective of severity/CTCAE grade. 
• any CTCAE grade 1 or 2 AE that raises a safety concern and is deemed related to 
IMP. 
The resumption of the trial can only occur after an amendment to the protocol has been 
approved by the HREC.  
4.3.8.3 
Special Individual Toxicity Rules 
Dosing of IMP in a participant will be suspended if they experience any of the following: 
• LFT/Transaminase elevation: 
o ALT or AST value >3× ULN together with: 
o Total bilirubin increase >2× ULN OR  
o INR >1.5 (see Sections 7.4.1.2 and 7.4.1.3). 
o ALT or AST value >8× ULN. 
o ALT or AST value >3× ULN, and with the appearance of fatigue, nausea, 
vomiting, right upper quadrant pain or tenderness, fever, rash, and/or 
eosinophilia > ULN. 
• Haematology: 
o Haemoglobin decrease from baseline (Day -1) of 4g/dL (40g/L) or lower. 
o Clinically significant decrease from Day -1 in neutrophil count (grade 3 [i.e. 
Absolute count < 1 x 109/L (1000/mm3)] or higher). 
o Platelet count drop of >50% from Day -1, or absolute value < 125 x109/L. No 
dose reduction of IMP permitted.  
4.3.8.4 
Special Group Toxicity Rules 
• If one participant fulfils the special individual toxicity rules, and this could be 
reasonably attributed to IMP, dosing for all participants in the trial will be 
temporarily suspended and a meeting arranged with the Principal Investigator, 
Sponsor Medical Director and study Medical Monitor (at a minimum) to decide if  
dosing will continue for the remaining participants. 
• If two or more participants fulfil any of the other special individual toxicity rules, 
then dosing will be suspended in all participants and can only continue if a  meeting 
with the Principal Investigator, Sponsor Medical Director and study Medical 
Monitor (at a minimum) is arranged (and a decision made that dosing can continue), 
and a substantial amendment is made to the protocol which has been 
reviewed/approved by the HREC. 
4.3.9 Emergency Unblinding of Study Participants 
• Emergency unblinding is not required for this study, as the Principal Investigator or 
delegate and Medical Monitor will be aware of each participant’s treatment. The study 
is single-blinded, so that only the participant is unaware of their treatment.  


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5 TRIAL INTERVENTIONS/TREATMENTS 
5.1 
INVESTIGATIONAL MEDICINAL PRODUCT (IMP) 
Participants will be admitted to the clinical unit for oral dosing of the combination of AL 
and Rux or AL and placebo as applicable.  AL will be administered first, and Rux or 
placebo (where applicable) administered 2 hours later, b.i.d. for 3 consecutive days.  
NOTE:  
• See Section 6.2 for permitted time windows when IMP administration coincides 
with PK or PD sampling. Scheduled times of IMP administration should be strictly 
adhered to and actual times will be documented and entered into the eCRF.  
• Participants will be confined to the clinical unit for all doses for a period of at least 
78 hours post-first administration of AL.  
• Participants will be administered all IMP doses under direct observation of clinical 
unit staff at the scheduled timepoints.  
• Participants will be blindfolded for administration of Rux/placebo, and suitable 
arrangements made to prevent other participants from witnessing each other’s 
Rux/placebo dosing. 
• For the first dose of IMP, and then for all doses that coincide with other assessments: 
participants will remain seated for 10 minutes, then stay in a semi-recumbent 
position for the next hour (except during assessments that require lying positions). 
5.1.1 Artemether-lumefantrine (Riamet®) 
• A course of Riamet® will be administered to all participants.  
• Each AL tablet contains 20 mg artemether and 120 mg lumefantrine.  
• The standard adult dosing regimen according to the Product Information and/or CMI 
will be used: 4 tablets administered orally b.i.d over 3 consecutive days (6 doses at 
t = 0, 8, 24, 36, 48 and 60 hours, total course of 24 tablets).  
• Each AL dose will be taken with 250 mL full-fat milk.  
• The first AL tablet will be administered under direct observation at time (t)  = 0  on 
Day 1 to all Part 1 participants and thereafter under direct observation at t = 8, 24, 
36, 48 and 60 hours.  
• No food permitted within 30 minutes prior to AL dosing, or between AL and Rux 
dosing. Meals are not required to be standardized. Times of meals and completion 
times will be documented.   
5.1.2 Ruxolitinib (Jakavi®) 
• A course of Rux (Jakavi®) in the form of 20 mg tablets will be administered only to 
the 6 participants randomised to the active group: 1 participant in Group 1a and 5 
participants in Group 1b.   
• Participants must be blindfolded for Rux/placebo administration; other participants 
will be prevented from witnessing dosing.  
• The equivalent of the standard adult dosing regimen according to the PI and CMI 
will be used: 1 tablet (1× 20 mg) administered orally with 250 mL water b.i.d over 
3 consecutive days (6 doses, total course of 6 tablets) and will be taken 2 hours after 
administration of AL. Participants will be instructed to swallow the IMP whole 
without biting or chewing.  
• The first 20 mg dose of Rux will be administered under direct observation at t = 2 
hours, and thereafter under direct observation at t = 10, 26, 38, 50 and 62 hours. All 
doses must be two hours after the corresponding dose of AL.  


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• No food permitted between AL and Rux, and for at least 1 hour post-Rux. Meals are 
not required to be standardized. Times of meals and completion times will be 
documented. 
5.1.3 Ruxolitinib Placebo 
• A placebo in capsule form will be administered only to the 2 participants randomised 
to placebo: 1 participant in Group 1a and 1 participant in Group 1b.  
• Participants must be blindfolded for Rux/placebo administration; other participants 
will be prevented from witnessing dosing.  
• A course of placebo treatment will be the same dosing regimen and food restrictions 
as described for Rux in Section 5.1.2. Participants will be instructed to swallow the 
IMP whole without biting or chewing. 
• Refer to the Pharmacy Manual for further information. 
5.2 
SELECTION OF DOSE IN THE STUDY  
See Section 2.3.2.  
5.3 
SELECTION AND TIMING OF DOSE FOR EACH PARTICIPANT 
See Section 5.1.  
5.4 
DOSE INTERRUPTIONS AND REDUCTIONS  
IMP administration may be interrupted/suspended under the conditions outlined above in 
Section 4.3.8. Otherwise, dose reductions are not permitted for AL or Rux during 
treatment.  
5.5 
SUPPLY, PACKAGING AND LABELLING OF STUDY TREATMENTS 
The IMPs will be manufactured and packaged according to Good Manufacturing Practice 
(GMP) and all local regulations, and will be labelled for clinical trial use in accordance 
with Australian requirements and Annex 13 GMP. Rux (Jakavi®) tablets, placebo capsules 
and AL (Riamet®) tablets will be supplied to the clinical unit with an acknowledgement 
of receipt form.  
Investigational Medicinal Products 
Riamet® (artemether-lumefantrine [AL]) 
Riamet® tablets (each containing 20 mg artemether/120 mg lumefantrine) will be supplied 
as  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. 
Jakavi® (ruxolitinib [Rux]) 
Jakavi® tablets (containing 20 mg ruxolitinib) will be supplied as elongated, curved white 
to almost-white tablets with ‘L20’ on one side and ‘NVR’ on the other. Each pack contains 
56 tablets.   
Ruxolitinib (Rux) placebo 
Placebo capsules will be prepared by PCI Melbourne and filled with microcrystalline 
cellulose (MCC) only.   
PCI Melbourne will fill all placebo capsules into one bulk container. A single panel 
label will be applied. The Principal Investigator, pharmacist or delegate will dispense 
the placebo capsules.  


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5.6 
STORAGE OF STUDY TREATMENTS 
Prior to dispensing, IMPs must be stored in a secure and locked storage area with limited 
access, and under monitored, temperature controlled conditions as appropriate.  
The Pharmacist or Designee will be responsible for the correct storage and handling of 
IMPs. Deviations from the storage requirements, including corrective action, must be 
documented.   
All storage requirements are detailed in the product information/CMI.  
5.7 
ACCOUNTABILITY, RECONCILIATION AND RETURN OF THE STUDY 
TREATMENTS 
The Principal Investigator or delegate will only dispense the IMPs to eligible participants 
enrolled in this study and must maintain complete and current dispensing and inventory 
records. All dispensing episodes for IMPs will be documented on the site’s Dispensing 
Logs. 
The logs must contain the following information: 
• Date of receipt. 
• Number of capsules/ tablets / containers received. 
• Batch number(s). 
• The identification of the participant to whom the capsules / tablets was dispensed. 
• The date(s), time and quantity dispensed to the participant. 
• The cumulative total of IMPs at site. 
• Capsules /tablets / containers damaged, destroyed, or returned. 
Supplies of IMPs will be shipped to the clinical unit prior to study start. The Monitor will 
complete drug accountability during routine monitoring visits.  
At the completion or termination of the study, final accountability and reconciliation will 
be completed and any discrepancies must be investigated and their resolution 
documented.  All full, partially full, and empty containers of IMPs must be returned to the 
Pharmacy, Manufacturer or Sponsor for destruction, and the appropriate form sent to the 
Sponsor.  
Please refer to the Pharmacy Manual for further details on the storage, handling, and 
dispensing of IMPs. 
5.8 
PROHIBITED CONCOMITANT THERAPY 
Permitted and prohibited prior and concomitant prescribed medications, over-the-counter 
medications and supplements are outlined in the inclusion and exclusion criteria in 
Section 4.3.1. Before use of any non-study medication during the study, the participant 
should discuss with the Principal Investigator or delegate.  
The following medications are prohibited during the study: 
• Psychiatric 
medications 
including 
antipsychotics 
(typical 
and 
atypical), 
antidepressants, mood stabilisers, and anticonvulsants. 
• Illicit drugs (amphetamine, cocaine, cannabis etc.). 
• Systemic or inhaled corticosteroids. 
• Anti-inflammatory drugs (excluding ibuprofen as indicated below) 
• Immunomodulators  


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• Immunosuppressive therapy (including systemic steroids, adrenocorticotrophic 
hormone or inhaled steroids) at a dose or duration potentially associated with 
hypothalamic-pituitary-adrenal axis suppression. 
• Anticoagulants. 
• Any vaccination (live or attenuated). 
• In consideration of specific ruxolitinib and Riamet® interactions:  
o CYP3A4 inhibitors or inducers, such as, but not limited to: boceprevir, 
clarithromycin, indinavir, itraconazole, ketoconazole, lopinavir/ritonavir, 
ritonavir, mibefradil, nefazodone, nelfinavir, posaconazole, saquinavir, 
telaprevir, telithromycin, voriconazole, avasimibe, carbamazepine, 
phenobarbital, phenytoin, rifabutin, rifampin (rifampicin), St.John's wort 
(Hypericum perforatum), ciprofloxacin, erythromycin, amprenavir, 
atazanavir, diltiazem, cimetidine, oxcarbazepine, barbiturates, efavirenz, 
nevirapine, pioglitazone, troglitazone, corticosteroids by the systemic 
route, grapefruit or grapefruit-containing products;  
o Dual inhibitors of CYP2C9 and CYP3A4 enzymes (e.g. fluconazole).  
• In consideration of specific Riamet® interactions:  
o Interaction with drugs that are known to prolong the QT, QTcF and/or 
QTcB interval: antiarrhythmics of classes IA and III, neuroleptics and 
antidepressant agents, certain antibiotics including some agents of the 
following classes: macrolides, fluoroquinolones, imidazole, and triazole 
antifungal agents, certain non-sedating antihistamines (terfenadine, 
astemizole), cisapride, flecainide;  
o Interaction with drugs metabolized by CYP2D6 since lumefantrine was 
found to inhibit CYP2D6 in vitro: (e.g. neuroleptics, metoprolol, and 
tricyclic antidepressants such as imipramine, amitriptyline, clomipramine) 
is contraindicated;  
o Antimalarials such as mefloquine and quinine;  
o Drugs that have a mixed inhibitory/induction effect on CYP3A4, 
especially anti-retroviral drugs such as HIV protease inhibitors (Lopinavir/ 
ritonavir) and non-nucleoside reverse transcriptase inhibitors (Efavirenz 
and Nevirapine).  
If a medication or treatment is administered that is in breach of these restrictions, the study 
Medical Monitor must be promptly notified in order to assess the participant’s eligibility 
for continued study participation. 
5.9 
PERMITTED CONCOMITANT MEDICATION 
The participant should discuss use of any medication during the study with the Principal 
Investigator or delegate.  
• The Principal Investigator or delegate may permit the use of ibuprofen up to 1.8 
g/day or paracetamol up to 1.2 g/day for the treatment of headache or other pain if 
required. Ibuprofen is the preferred treatment for headache or pain.  
5.10 TRIAL INTERVENTION/TREATMENT COMPLIANCE  
Participants will be confined to the clinical unit to be administered all doses of IMP under 
direct observation, and compliance will be confirmed and documented by clinical unit 
staff.   


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5.11 MEASURES TO MINIMIZE BIAS 
5.11.1 Randomisation Procedures 
Randomisation procedures will be as described in the Randomisation Plan and 
Randomisation Procedure.  
A randomisation number will be allocated to each participant as per the randomisation 
schedule before the administration of IMPs (AL+Rux or AL+placebo).  
Overall, randomisation will be in a ration of 3:1 (active:placebo): 
• Participants in Group 1a will be randomised to a treatment allocation (AL+Rux or 
AL+placebo) in a ratio of 1:1. The randomisation will be single blinded (treatment 
known to Investigator but not participant). 
• Participants in Group 1b will be randomised to a treatment allocation (AL+Rux or 
AL+placebo) in a ratio of 5:1. The randomisation will be single blinded (treatment 
known to Investigator but not participant).  
5.11.2 Blinding and Unblinding 
The study will be single-blinded, with only the participant unaware of their treatment 
allocation. The single-blind design is aimed at optimising participant safety by allowing 
the Principal Investigator or delegate to rapidly detect and manage potential safety 
concerns that may disproportionately arise in the combination AL+Rux arm.  
See Section 4.3.9. To maintain the blind for the participant, all source documentation will 
be prepared as blinded, participants will be blind-folded for administration of Rux/placebo 
and arrangements made to ensure participants cannot witness each other’s Rux/placebo 
dosing. A participant’s treatment allocation should not be unblinded to them except if 
required in a medical emergency.  
6 STUDY PROCEDURES AND ASSESSMENTS  
6.1 
STUDY CONDUCT SCHEDULE 
The study will be conducted according to the Schedule of Activities (see Section 1.3: 
Table 1, Table 2 and Table 3) and as outlined in this section. See Section 6.2 for permitted 
time windows for pharmacodynamic and PK sampling and Sections 6.3 and 6.4 for details 
on PD and PK sampling respectively. See Section 7 for details of safety assessments.  
6.1.1 Screening Days -28 to Day -1  
See Schedule of Activities Table 1. The potential participant will be asked to attend a 
screening visit having fasted for at least 8 hours for screening biochemistry testing. The 
initial Screening visit will be prior to and separate from the safety/eligibility visit on Day 
-1.  
Procedures at Screening: 
• Obtain signed informed consent prior to any study-related procedures or 
assessments.  
• Medical history 
• Current and prior medications, including details of recreational drug, alcohol and 
tobacco use 
• Demographics 
• Body measurements (weight and height) 


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• Alcohol breath test 
• Collect urine for: 
o Urine drug screen 
o Urinalysis  
• Full physical examination 
• Beck Depression Inventory (BDI-II) 
• Vital signs (supine and standing blood pressure and heart rate)  
• Tympanic body temperature and respiratory rate 
• 12-lead ECG  
• Collect blood samples for: 
o Haematology  
o Biochemistry (including lipids for Screening visit only) 
o Serology  
o Coagulation  
o QuantiFERON-TB Gold assay (tuberculosis testing) 
o Serum β-hCG pregnancy test for all females. 
o Follicle stimulating hormone (FSH) for postmenopausal females. 
• Verify eligibility.  
6.1.1.1 
Review of screening/eligibility laboratory testing  
• If results are normal/within the laboratory normal reference ranges, the participant 
may be included (if all inclusion criteria fulfilled and no other exclusion criteria are 
met).  
• If results are abnormal or outside of the laboratory normal reference ranges, then 
clinical significance must be assigned by the Principal Investigator or delegate:  
o If clinically significant, the participant is excluded as per Exclusion 
criterion #2.  
o If not clinically significant, the parameter(s) must be checked against 
Appendix 1: Clinically Acceptable Ranges for Clinically Important Study 
Inclusion/EXCLUSION Laboratory Tests:  
▪ If outside the ranges in Appendix 1, the participant is excluded as 
per Exclusion criterion #2.  
▪ If within the ranges in Appendix 1, the participant may be included 
(if all inclusion criteria fulfilled and no other exclusion criteria are 
met).  
• Retesting of any screening parameters (except urine drug screen) is permitted once 
only if the original result did not reflect the expected medical condition of the 
participant.  
o If a participant’s urine drug screen returns a positive result (except for 
paracetamol as per Exclusion criterion #34), no retest is permitted and the 
participant is excluded.  
• The results of any retests must be considered for participant eligibility and must be 
available prior to first-dose of IMP. Findings made during unscheduled visits or 
testing should be reported in the source document and eCRF.  
6.1.2 Eligibility/Safety/Admission Day -1  
See Schedule of Activities Table 1. Participants will be requested to attend the clinical 
unit to confirm eligibility and to be admitted to the clinical unit on Day -1 after fasting for 


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at least 8 hours. Reserve participants will also attend. This will be in addition to the 
Screening visit and should not be combined.  
Procedures on Day -1 for safety, to confirm eligibility and for admission: 
• Review medical history and prior/concomitant medications since screening visit. 
Record any medically untoward events between Screening and this visit as medical 
history.  
• Conduct alcohol breath test.  
• Collect urine for: 
o Urinalysis. Results must be available and reviewed as described in Section 
6.1.1.1 prior to IMP administration on Day 1.  
o Urine drug screen  
o Urine β-hCG pregnancy test for WOCBP 
• Participants may be cannulated with indwelling intravenous (IV) cannula either on 
Day -1 or pre-dose on Day 1.  
• Collect blood samples for: 
o Haematology 
o Biochemistry.   
o NOTE: Results must be available and reviewed prior to IMP 
administration on Day 1 as described in Section 6.1.1.1.  
• Abbreviated physical examiniation including weight (kg).  
• Vital signs (supine and standing blood pressure and heart rate)  
• Tympanic body temperature and respiratory rate 
• Randomisation may be conducted on Day -1 or Day 1 pre-first dose.  
 
6.1.3 Days 1-4 Treatment with IMP and Confinement 
See Schedule of Activities Table 2. Participants will fast overnight at the clinical unit for 
at least 8 hours. Eligibility must be confirmed prior to IMP dosing.  
NOTE: when several procedures are scheduled to take place at the same timepoint, the 
following order is recommended where applicable: urine collection, ECG, vital signs, 
blood sampling, IMP administration.  
6.1.3.1 
Day 1 Pre-first dose IMP 
Procedures prior to first dose of IMP (AL at t = 0): 
• Confirm eligibility (including review of all Day -1 safety laboratory testing and 
available Day 1 pre-dose testing) 
• Medical history (medically untoward events prior to first dose IMP will be recorded 
as medical history and must satisfy relevant inclusion/exclusion criteria) 
• Concomitant/prior medications 
• Vital signs (supine and standing blood pressure and heart rate) within 60 minutes 
prior to IMP dosing. Pre-dose results must satisfy relevant inclusion/exclusion 
criteria. 
• Tympanic body temperature and respiratory rate within 60 minutes prior to IMP 
dosing. Pre-dose results must satisfy relevant inclusion/exclusion criteria. 
• 12-lead standard ECG. Pre-dose results must satisfy relevant inclusion/exclusion 
criteria. 
• Cannulate with indwelling intravenous (IV) cannula (if not done on Day -1) 


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o Collect blood for pre-dose PK sample within 60 minutes prior to IMP 
dosing (see Section 6.4).  
o Collect blood for pre-dose pSTAT3 sample within 60 minutes prior to IMP 
dosing 
• Randomisation (if not done on Day -1). 
6.1.3.2 
IMP dosing 
Commencing Day 1, AL+Rux or AL+placebo will be administered under observation 
according to randomisation schedule. See Section 5.1 for details including timing and 
food/liquids/positioning requirements.  
Rux/placebo must be administered 2 hours after the corresponding AL dose.  
6.1.3.3 
Confinement 
Participants will be confined for at least 78 hours post-first dose IMP for safety and 
tolerability monitoring.  
Confinement Procedures: 
• Record AEs 
• Record concomitant medications 
• Collect blood samples for: 
o PK – for timepoints coinciding with IMP administration, PK sampling 
must occur prior to IMP administration - see Section 6.4 below.  
o pSTAT3 assay. For timepoints, see Section 6.3.1 below.  
o Safety biochemistry and haematology blood sampling within 60 minutes 
prior to 24, 48 and 72 hours after first dose of IMP (must be before 
breakfast). Participants will have fasted for at least 8 hours prior to blood 
sampling.  
• Collect urine for urinalysis within 60 minutes prior to 24, 48 and 72 hours after first 
dose of IMP.    
• Vital signs (supine blood pressure and heart rate)  at 4 and 8 hours after first dose of 
IMP and then every 8 hours (within 15 minutes prior to each timepoint).  
• Tympanic body temperature and respiratory rate (supine) at 4 hours post-first dose 
of IMP and then 24, 48 and 72 hours post-first dose IMP (within 15 minutes prior to 
each timepoint).   
• Standard single 12-lead ECG at 24, 48 and 72 hours after first dose of IMP (within 
15 minutes prior to each timepoint).  
• Symptom-directed physical examination at any time if clinically indicated at the 
discretion of the Principal Investigator or delegate. 
Prior to discharge 78 hours post-first dose IMP:  
• Collect blood sample for PK at t = 78 hours timepoint as per Section 6.4 
• Record AEs and concomitant medications 
• Clinical unit staff will check with the Principal Investigator or delegate to clarify if 
symptom-directed physical examination required prior to discharge.  
6.1.4 Discharge to EOS Outpatient Visits  
After discharge from the clinical unit 78 hours post-first dose IMP, participants will attend 
outpatient visits at the clinical unit scheduled for safety assessments and to ensure blood 
sampling for PK and pSTAT3 assay will be taken at the required timepoints (see Schedule 
of Activities Table 3).  


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Outpatient Visit procedures: 
Collect blood for: 
• PK samples– see Section 6.4. 
• pSTAT3 samples – see Section 6.3.1. 
At all visits (time windows for assessments are as permitted for PK sampling in Section 
6.2): 
• Record AEs 
• Record concomitant medications 
• Vital signs (supine blood pressure and heart rate only)  
• Tympanic body temperature and respiratory rate  
• Symptom-directed physical examination if clinically indicated 
On Day 8 and Day 15 only (participants should attend having fasted for at least 8 hours): 
• Collect blood for: 
o Haematology 
o Biochemistry  
• Collect urine for urinalysis  
• Standard single 12-lead ECG 
 
6.1.5 End of Study Visit (EOS) Day 29 or Early Termination Visit 
Participants will be requested to attend the EOS (or early termination visit if applicable) 
having fasted for at least 8 hours. EOS assessments must be conducted within the 
permitted time window for the applicable PK sample (see Section 6.2).  
EOS (or Early Termination) procedures: 
• Record AEs 
• Record concomitant medications 
• Full physical examination 
• Vital signs (supine blood pressure and heart rate only)  
• Tympanic body temperature and respiratory rate 
• 12-lead ECG  
• Collect blood samples for: 
o PK (if applicable; see Section 6.4) 
o Haematology 
o Biochemistry  
o serum β-hCG pregnancy test for WOCBP 
• Collect urine for urinalysis 
 
 


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6.2 
PERMITTED TIME WINDOWS FOR SAMPLING 
The following time windows will be permitted for blood sampling for PK and pSTAT3 
assays: 
Time point  
Tolerance window 
Pharmacokinetic/Pharmacodynamic (pSTAT3) 
In confinement 
Pre-first dose IMP  
- 60 min to 0 h 
1-4 
hours 
inclusive 
after 
initial 
AL 
administration  
(T 2 h must be within 2 min pre Rux/placebo 
dosing) 
± 2 min 
5-12 
hours 
inclusive 
after 
initial 
AL 
administration (T 8h must be within 5 min pre 
AL dosing) 
± 5 min 
13-78 
hours 
inclusive 
after 
initial 
AL 
administration (T 24, 36, 48 & 60 h must be 
within 15 min pre AL dosing) (T 62 h must be 
within 15 min pre Rux/placebo dosing) 
± 15 min 
Outpatient Visits 
79-168 hours inclusive (pSTAT3 assay T 168 h) ± 120 min 
169-EOS  
± 24 h 
 
6.3 
PHARMACODYNAMIC ASSESSMENTS 
6.3.1 pSTAT3  
Blood samples will be collected as described in the Laboratory Manual to determine the 
levels of pSTAT3 using a validated whole blood enzyme-linked immunosorbent assay 
(ELISA). See Section 6.2 for permitted time windows.  
For timepoints coinciding with IMP administration, blood sampling MUST occur prior 
to IMP administration. Exact times of pSTAT3 sampling will be documented and entered 
into the eCRF.  
Blood samples for pSTAT3 assay will be collected at the following times: 
• Day 1  prior to commencement of IMP dosing (pre-dose). 
• t = 1, 2, 3, 4, 5, 6, 8, 12 hours post commencement of IMP dosing. 
• Day 8 at 168 hours post commencement of IMP dosing. 
6.4 
PHARMACOKINETICS 
Plasma concentrations of Rux, AL and artemether metabolite dihydroartemisinin (DHA) 
will be quantified using a validated method by liquid chromatography-mass spectrometry 
(LC MS/MS). 
Blood sample handling for PK analysis is described in the Laboratory Manual. Blood 
samples will be collected either by direct venepuncture or via an indwelling cannula 
inserted in a forearm vein. See Section 6.2 for permitted time windows. Exact times of 
PK sampling will be documented and entered into the eCRF. 
Blood samples for PK will be collected at the following times: 


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• Day 1 prior to commencement of IMP dosing (pre-dose). 
• t = 1, 2, 3, 4, 5, 6, 8, 12, 24, 36, 48, 60, 62, 64, 72 and 78 hours post commencement 
of IMP dosing. 
o For timepoints coinciding with IMP administration, blood sampling 
MUST occur prior to IMP administration. 
• Days 8, 11, 15, 21, 24 and 27 at t = 168, 240, 336, 480, 552 and 624 hours post 
commencement of IMP dosing (±120 min for t = 168 hours, ± 24 hours for later 
timepoints). 
• Day 29 at t = 672 hours (± 24 hours).  
• Unscheduled PK sample will be taken if ALT or AST value >2x ULN as described 
in Section 7.4.1.2. 
7 SAFETY MONITORING 
• Safety will be assessed during the study through physical examination, urinalysis, 
vital signs, ECG, clinical laboratory tests, and continuous AE reporting, as outlined 
in the Schedules of Activities (Section 1.3). 
• Protocol waivers or exemptions are not permitted.  
• Clinical significance of any out-of-range or abnormal result or finding must be 
recorded by the Principal Investigator or delegate. After first-dose of IMP, any 
clinically significant observation for safety monitoring (physical examination, vital 
signs, ECG, clinical laboratory tests) as determined by the Principal Investigator or 
delegate must be documented as an AE (see Section 7.6.1).   
• See Section 6.1.1.1 for review of results for Screening/eligibility purposes.  
• All assessments may be performed at extra unscheduled times if deemed necessary 
by the Principal Investigator or delegate to ensure participant safety. The Medical 
Monitor may be contacted if required.  
• NOTE: when several procedures are scheduled to take place at the same timepoint, 
the following order is recommended where applicable: urine collection, ECG, vital 
signs, blood sampling, IMP administration. 
7.1 
PHYSICAL EXAMINATION 
7.1.1 Full Physical Examination 
A full physical examination will include assessments of general appearance; head, neck 
(including thyroid), ears, nose and throat; heart/circulation; chest; lungs; abdomen; skin, 
and a neurological examination. 
A full physical examination will be performed at Screening and at the EOS.  
7.1.2 Abbreviated Physical Examination 
An abbreviated physical examination will include heart/circulation, chest, lungs, skin, and 
abdomen, and will be performed on Day -1 on admission to the clinical unit. This will 
include re-checking the weight (kg) of participants.  
7.1.3 Symptom Directed Physical Examination 
A symptom-directed physical examination may be performed at any time during the study 
if clinically indicated, and body systems to be reviewed will be only as clinically 
indicated. Clinical unit study staff must ensure a study doctor is contacted if there are 
symptoms that require physical examination, including prior to discharge from the clinical 
unit after confinement. 


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7.2 
VITAL SIGNS AND OTHER BODY MEASUREMENTS 
The normal ranges for vital signs, body temperature and respiratory rate once on trial are: 
Parameter 
Range 
Systolic blood pressure             
90-140 mmHg 
Diastolic blood pressure           
50-90 mmHg 
Heart rate                                 
50-100 bpm 
Tympanic body temperature                             
35.0-37.5°C 
Respiratory rate                        
10-25 breaths/min 
If results are out of these ranges, the Principal Investigator or delegate must assess, assign 
and document clinical significance (see Section 7.6.1).  
7.2.1 Vital signs 
Vital signs include systolic and diastolic blood pressure and heart rate. Systolic blood 
pressure (SBP) and diastolic blood pressure (DBP) will be measured in millimeters of 
mercury (mmHg) and heart rate in beats per minute.  
Blood pressure and heart rate will be measured after the participant has rested in the supine 
position for at least 5 minutes and then in the standing position within 2–3 minutes when 
changing from the supine to standing position at Screening, Day -1 and Day 1 within 60 
minutes of the first dose of IMP. All pre-first dose results including Day 1 pre-first dose 
results must satisfy relevant inclusion/exclusion criteria. 
At all other timepoints, vital signs will be measured after the participant has rested in a 
supine position for at least 5 minutes: during confinement at 4 and 8 hours post-first dose 
of IMP and then every 8 hours (within 15 minutes prior to each timepoint), and at every 
outpatient visit after discharge from the clinical unit and EOS (time windows as per PK 
sampling in Section 6.2).   
7.2.2 Other Body Measurements – Respiratory rate and Temperature 
Respiratory rate (breaths per minute) and tympanic body temperature (°Celcius) will be 
measured at Screening, Day -1, Day 1 pre-dose (within 60 minutes), at 4, 24, 48 and 72 
hours post-first dose of IMP (within 15 minutes prior to timepoint), then at every 
outpatient visit after discharge from the clinical unit and EOS (time windows as per PK 
sampling in Section 6.2).  Day 1 pre-first dose results must satisfy any relevant 
inclusion/exclusion criteria. 
7.2.3 Other Body Measurements – Weight and Height 
Weight (kg) and height (cm) will be measured at Screening, and weight (kg) will be re-
checked on Day -1. 
7.3 
12-LEAD ECGS 
Single, standard 12-lead ECGs will be recorded after the participant has rested supine for 
at least 5 minutes. ECG tracings will be retained and labelled as per standard procedures 
at the clinical unit and recorded in the eCRF. The Principal Investigator or delegate will 
sign and date each ECG as evidence of their review. 
ECGs will be taken at Screening,  within 60 minutes of the first dose of IMP on Day 1, 
then 24, 48 and 72 hours post-first dose IMP (within 15 minutes prior to each confinement 
timepoint), Day 8, Day 15 and at EOS (outpatient time windows as per PK sampling). 
Day 1 pre-first dose results must satisfy relevant inclusion/exclusion criteria. 


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Normal ECG ranges once on trial are: 
Parameter 
Range 
PR interval             
≤210 msec 
QRS           
50–120 msec 
QTcB/QTcF                            
≤450 msec 
If results are out of these ranges, the Principal Investigator or delegate must assess and 
assign clinical significance (see Section 7.6.1).  
7.4 
CLINICAL LABORATORY TESTS 
Handling of blood and urine sampling for laboratory safety assessments is described in 
the Laboratory Manual and/or the preferred Vendor Pathology Laboratory Manual. 
Biochemistry, haematology and urinalysis will be analysed by the clinical unit’s preferred 
Vendor pathology laboratory. Urine pregnancy testing, urine drug screening and alcohol 
breath testing will be conducted at the clinical unit. 
See Section 6.1.1.1 for review of results for Screening/eligibility purposes.  
7.4.1 Biochemistry and Haematology Safety Laboratory Testing 
• When biochemistry and haematology safety laboratory tests are scheduled, 
participants should attend the clinical unit having fasted for at least 8 hours. During 
confinement, clinical unit staff should ensure participants have fasted for at least 8 
hours prior to all scheduled safety laboratory sampling.   
• The Principal Investigator or delegate must review all results and assign clinical 
significance to any results that are abnormal and/or outside the laboratory normal 
reference ranges:  
o For Screening and eligibility, participants will be excluded if they return 
safety laboratory test results outside of the laboratory normal reference 
ranges AND the results are: 
▪ considered not clinically significant by the Principal Investigator 
but are also outside the ranges specified in Appendix 1: Clinically 
Acceptable 
Ranges 
for 
Clinically 
Important 
Study 
Inclusion/EXCLUSION Laboratory Tests, OR  
▪ considered clinically significant by the Principal Investigator.   
o After IMP is commenced, results must also be reviewed with reference to 
the toxicity rules for this study (see Section 4.3.8) and AESIs (see Section 
7.6.5). See also Section 7.6.1 for clinically significant results. 
• Results of safety laboratory tests conducted on Day -1 must be available and 
reviewed prior to first administration of IMP on Day 1, and must satisfy relevant 
inclusion/exclusion criteria.  
• Blood samples for haematology and biochemistry laboratory safety assessments  
will be collected at Screening (including blood group and Rh(D), lipids), Day -1, at 
24, 48 and 72 hours after first-dose IMP (within 60 minutes prior to each 
confinement timepoint), Day 8, Day 15 and EOS (outpatient time windows as per 
PK sampling in Section 6.2).  
7.4.1.1 
Haematology Parameters 
Haematology: Full blood count (FBC) with differential, white blood cell count (WBC) 
with differential (neutrophils, lymphocytes, monocytes, eosinophils, basophils), Mean 


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corpuscular volume (MCV), red blood cell (RBC) count, haemoglobin, haematocrit, 
platelet count, red blood cell distribution width,  and reticulocyte count. Blood group and 
Rh(D) tests to be included at Screening only.  
• 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.  
• Blood will be also be collected for Haematology if ALT or AST >2x ULN as 
described in Section 7.4.1.2. 
7.4.1.2 
Biochemistry Parameters 
Biochemistry: Albumin, alkaline phosphatase (ALP), alanine aminotransferase (ALT), 
aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), Lactate 
dehydrogenase (LDH), bicarbonate, total and direct bilirubin, urea, calcium, corrected 
calcium, phosphate, sodium, creatinine, estimated glomerular filtration rate (eGFR; 
Chronic Kidney Disease Epidemiology collaboration [CKD-EPI equation]), glucose 
(fasted), potassium, chloride, magnesium, uric acid, total protein, creatine kinase (CK);  
• Lipid profile (at Screening only): total cholesterol, triglycerides, high density 
lipoprotein (HDL), low density lipoprotein (LDL) [participants must fast for at least 
8 hours prior] 
• If ALT or AST value >2 ULN post-first dose IMP, then: 
o  blood samples for the following tests should be collected within 24-48 
hours:  
▪ Coagulation (see Section 7.4.1.3; also Section 7.6.5 and Section 
4.3.8.3)  
▪ Liver panel (ALT, AST, LDH, ALP, GGT, total and direct 
bilirubin)  
▪ CK 
▪ Haematology (see Section 7.4.1.1)  
▪ Unscheduled PK sample.  
o Collect additional history of potentially hepatoxic concomitant 
medications/substances (including paracetamol and alcohol) and exercise. 
7.4.1.3 
Coagulation 
Coagulation at Screening only; may also be requested at any time during the study for 
safety at the discretion of the Principal Investigator or delegate, and must be requested if 
ALT or AST > 2x ULN as described in Section 7.6.5 and Section 4.3.8.3: 
• prothrombin time (PT), activated partial thromboplastin time (APTT), international 
normalised ratio (INR)  
7.4.2 Urinalysis Safety Laboratory Testing 
• Urine will be collected to conduct urinalysis at Screening, Day -1 Eligibility, at 24, 
48 and 72 hours after first-dose IMP (within 60 minutes prior to each confinement 
timepoint), Day 8, Day 15 and EOS (with biochemistry and haematology safety 
blood testing; outpatient time windows as per PK sampling).  
o Dipstick testing on a freshly voided urine sample including tests for 
proteins, glucose, blood (erythrocytes), leukocytes, ketone bodies, nitrite, 
bilirubin, urobilinogen, and pH.  
o Quantitative 
assessment 
of 
glucose, 
protein, 
creatinine 
and 
protein:creatinine ratio for each sample;  


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o Any urine dipstick test result of more than traces (pH will not trigger 
quantitative analysis) will trigger a quantitative analysis for all of the 
following parameters: red blood cells, white blood cells, and bacteria 
(bacteria not quantitative).  
o If available, the quantitative results will be the leading one over the 
dipstick results for decision making eg, for study inclusion/exclusion or 
clinical assessment.  
o See Section 7.4.1 for information on screening/eligibility review of results.  
Urinalysis will be conducted at the clinical unit’s preferred Vendor pathology laboratory. 
7.4.3 Pregnancy and Follicle Stimulating Hormone Testing 
• Follicle Stimulating Hormone (FSH): blood will be collected from postmenopausal 
females only at Screening. 
• Serum β-hCG pregnancy test: blood will be collected from all women during 
Screening and from WOCBP only at EOS. 
• Urine β-hCG pregnancy test: urine will be collected from all WOCBP on Day -1. 
o A positive urine pregnancy test must be confirmed by a serum β-hCG 
pregnancy test.  
7.4.4 Serology and Tuberculosis Testing 
• Serology: blood will be collected at screening  
o testing for hepatitis B (HBsAg, anti-HBc [IgG + IgM if IgG is positive]), 
hepatitis C (anti-HCV), human immunodeficiency virus (HIV) HIV 1/2 
(anti-HIV1 and anti-HIV2 Ab) 
• Latent M. tuberculosis infection testing using Quantiferon-TB gold assay: 
o  blood will be collected at Screening; 
o One repeat can be performed for indeterminate result. If repeat result is 
again indeterminate the individual will not be eligible.  
7.4.5 Alcohol and Drug Screening 
• Urine will be collected for drug screening at Screening and Day -1, testing for: 
o Acetaminophen/paracetamol (APAP) (not exclusionary) 
o Amphetamines (AMP) 
o Methamphetamines (mAMP) 
o Barbiturates (BAR) 
o Benzodiazepines (BZO) 
o Cocaine (COC) 
o Methadone (MTD) 
o Opiates (OPI) 
o Phencyclidine (PCP) 
o Cannabis (THC) 
o Tricyclic antidepressants (TCA)  
o 3,4-methylenedioxy-methamphetamine (MDMA) 
• Alcohol will be tested for by breath test at Screening and on Day -1.  
If considered necessary to confirm lifestyle considerations are being adhered to during the 
study (Section 4.3.3), alcohol and drug screening tests may also be conducted at other 


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unscheduled timepoints at the discretion of the Principal Investigator or delegate. 
However, re-tests for screening/eligibility purposes are not permitted.  
7.5 
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.38 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 for eligibility.39 The BDI-II takes approximately 10 minutes to complete, 
although participants require a fifth – sixth grade reading level to adequately understand 
the questions. A score of >20 at screening and/or a response of 1, 2 or 3 for item 9 
indicating current suicidal ideation is exclusionary (see exclusion criterion 12 Section 
4.3.2). A BDI score of 17 to 19 may be enrolled at the discretion of the Principal 
Investigator if they do not have a history of the psychiatric conditions mentioned in 
exclusion criterion 12 (Section 4.3.2) and their mental state is not considered to pose 
additional risk to the health of the volunteer or the execution of the study and 
interpretation of the data gathered. 
7.6 
ADVERSE EVENTS (AES) 
The Principal Investigator or delegate and clinical facility staff are responsible for 
detection, recording, and reporting of events that meet the criteria and definition of various 
adverse events (AEs) as listed below. Treatment emergent adverse events (TEAEs)  will 
be recorded from the administration of the first dose of IMP on Day 1 (see Schedule of 
Assessments Section 1.3).     
All individual events will be followed until: 
• The event is resolved, or 
• No further medically relevant information in relation to the event can be expected, 
and 
• the Principal 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 the Principal 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 study, if judged necessary. 
Any medically untoward event occurring in participants from the time of consent to the 
time of administration of IMPs will be considered medical history. 
7.6.1 Definitions 
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 study, and which does not necessarily have a causal 
relationship with study 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 study 
even though it may have been present prior to the start of the study. 
• An exacerbation of a pre-existing medical condition/disease. 


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• An increase in frequency or intensity of a pre-existing episodic disease or medical 
condition. 
• Continuous persistent disease or symptoms present at study start that worsen 
following the start of the study. 
• An abnormal assessment (e.g., change on physical examination, ECG finding) if it 
represents a clinically significant finding that was not present at study start or 
worsened during the course of the study. 
• 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 study 
start or worsened during the course of the study or led to dose reduction, interruption 
or permanent discontinuation of study treatment. 
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 (Section 7.7) or AESI (Section 
7.6.5); 
• 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 IMP dosing, or discontinuation from the study, significant 
additional concomitant drug treatment, or other therapy, and/or 
• It is considered by the Principal Investigator (or delegate) or Sponsor to be clinically 
significant or represent a clinically significant change from baseline. 
AEs will not include: 
• A medical or surgical procedure such as surgery, endoscopy, tooth extraction, or 
transfusion (although the condition that leads to the procedure may be an AE).  
• A pre-existing disease or condition present at the start of the study that does not 
worsen during the study. 
• Any situation where an untoward medical occurrence has not occurred (for example, 
hospitalizations for cosmetic elective surgery or social admissions). 
7.6.2 Causal Relationship to Investigational Medicinal Product  
A causal relationship of any adverse events experienced by participants to either of the 
IMPs will be determined by reference to the current product information and/or CMI.  
If considered related: 
• The temporal relationship between the event and the administration of IMPs 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.  
If considered not related: 
• The event can be readily explained by other factors such as the participant’s 
underlying medical condition, concomitant therapy or accident and no plausible 
temporal or biologic relationship exists between the challenge agent and/or IMPs 
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7.6.3 Severity Grading of Adverse Events 
The severity of AEs will be recorded in accordance with the CTCAE v5.0, published 27 
November 2017. This guidance provides a common language to describe levels of 
severity, to analyse and interpret data, to scale the aggregate AE score, and to articulate 
the clinical significance of all AEs. 
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. 
An AE that is assessed as severe should not be confused with a SAE. Severity is a category 
utilized for rating the intensity of an event; and both AEs and SAEs can be assessed as 
severe. An AE is defined as ‘serious’ when it meets one of the pre-defined serious 
outcomes as described in Section 7.7.1. 
7.6.4 Documentation of Adverse Events 
When an AE occurs, it is the responsibility of the Principal Investigator or delegate to 
review all documentation (e.g. progress notes, laboratory, and diagnostics reports) relative 
to the event. The Principal Investigator or delegate will then record the AE on the AE 
eCRF. Additional reporting requirements for an AE meeting serious criteria are discussed 
in Section 7.7.2 below. The Principal Investigator or delegate will attempt to establish a 
diagnosis of the event based on signs, symptoms, and/or other clinical information. In all 
cases, when available, the diagnosis should be reported as the event and not the individual 
signs/symptoms. 
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 
• Seriousness  
• Severity 
o In the source data, the description of the AE will report the various 
severities observed over time. If the severity of an AE increases, separate 
AEs per severity grading will be recorded into the eCRF. 
o If the AE resolves and then reoccurs, then two AEs will be reported. 
• Action taken in response to the AE regarding IMP: 
o Dose not changed  
o Drug interrupted  
o Drug withdrawn  
o Not applicable  
o Unknown 
• Outcome of AE: 


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o Recovered/resolved, or 
o Recovering/resolving, or 
o Not recovered/not resolved, or 
o Recovered with sequelae/resolved with sequelae 
o Fatal 
o Unknown  
• Relationship to the trial treatments or procedures (causality assessment of related or 
not related), or any other treatment or procedure conducted during the trial.  
7.6.5 Adverse Events of Special Interest 
An adverse event of special interest (AESI) (serious or non-serious) is one of scientific 
and medical concern specific to the Sponsor’s product or programme, for which ongoing 
monitoring and rapid communication by the PI to the Sponsor is appropriate. Such an 
event may require further investigation in order to characterise and understand it.  
The pharmacovigilance provider (Prime Vigilance) must be notified of all AESIs (serious 
and non-serious) within 24 hours of the site becoming aware of the event. The notification 
should be via a Serious Adverse Event Report Form (marked as ‘AE of special interest’). 
Follow-up information will be submitted in a timely fashion as further information 
become available. 
Hepatic AEs of Special Interest 
• Any ALT or AST above 5×ULN, 
• An elevation in bilirubin 2×ULN, 
• Any AST or ALT above 2×ULN and: 
o Total Bilirubin Level (TBL) >1.5×ULN OR  
o INR >1.4 (see Section 7.4.1.2), 
• Any AST or ALT 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). 
Cardiac AEs of Special Interest 
• QTcB or QTcF at any time >480 msec, 
• Bundle branch block (except right bundle branch block that was present prior to IMP 
administration), 
• Any arrhythmia, except: 
o Sinus bradycardia that is clinically asymptomatic, and not associated with 
any other relevant ECG abnormalities, 
o sinus tachycardia that is clinically asymptomatic, and associated with a 
body temperature >38.0°C, and not associated with any other relevant 
ECG abnormalities, 
o Respiratory sinus arrhythmia, 
o Wandering atrial pacemaker, 
o Isolated, single premature atrial/ventricular complex (i.e., no bigeminy, 
trigeminy, couplets, triplets or salvos) that does not occur more than once 
in a particular ECG tracing. 
Haematological AEs of Special Interest 
• Absolute neutrophil count <0.5 × 109/L, 
• Platelet count <125 × 109/L. 


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Dermatological AEs of Special Interest 
Clinical signs of possible cutaneous adverse reactions such as:  
• Dermatitis, 
• Rash, including erythematous, macular, papular, maculopapular, pruritic, pustular, 
and vesicular. 
If one of these cutaneous reaction is observed and when feasible, pictures of the lesions 
should be obtained. 
Dermatological AEs need not be reported as AESIs if clearly unrelated to IMP (e.g. rash 
from cannula dressing or ECG dots). 
7.7 
SERIOUS ADVERSE EVENTS (SAES) 
7.7.1 Definitions of Serious Adverse Events (SAEs) 
An SAE is any AE that: 
• Results in death, or 
• Is life-threatening, or 
• Requires inpatient hospitalization or prolongation of existing hospitalization, or 
• Results in persistent or significant disability/incapacity, or 
• Is a congenital anomaly/birth defect 
• Constitutes a possible Hy’s Law case: 
o Hy’s Law case is defined as a subject 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 not associated to an alkaline 
phosphatase value greater than 2×ULN (FDA Guidance on Drug Induced 
Liver Injury: Premarketing Clinical Evaluation [2009]). 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered SAEs when, based upon appropriate medical judgment, 
they may jeopardize the subject and may require medical or surgical intervention to 
prevent one of the outcomes listed in this definition. 
The term ‘life-threatening’ in the definition of ‘serious’ refers to an event/reaction in 
which the subject was at immediate risk of death at the time of the event/reaction; it does 
not refer to an event/reaction, which hypothetically might have caused death, if it were 
more severe.  
Any planned procedures that require admission to hospital for compliance with this 
protocol as well as any planned elective procedures are not considered SAEs (unless the 
underlying condition has worsened or the procedure results in a worsening of the subject’s 
condition).   
7.7.2 Reporting for Serious Adverse Events (24 hours) 
SAEs must be followed until their signs and symptoms have remitted or stabilized, until 
the participant is lost to follow-up, or until the End of Study Visit, whichever occurs last. 
AEs meeting the serious criteria MUST be reported promptly to the pharmacovigilance 
provider (Prime Vigilance) and as per the Safety Management Plan within 24 hours of the 
site becoming aware of the event:  
SAE Reporting: 
PrimeVigilance  


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Email: MMV@primevigilance.com 
Fax: +44 (0) 800 471 5694 
Any copies of participant’s medical records provided for SAE reporting must have all 
participant identifiers redacted before submission. 
The SAE report form 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 SAE, he/she 
will not wait to receive additional information before reporting the event. A follow-up 
SAE report should be completed within 14 days, or if there is no new information the 
SAE 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 as described in Section 7.6.2. 
Email transmission of the SAE report form is the preferred method to transmit this 
information to Prime Vigilance. In rare circumstances notification by telephone is 
acceptable, with a copy of the SAE report form sent by overnight mail.  
Initial notification via the telephone does not replace the need for the Principal 
Investigator or delegate to complete and sign the SAE report form within the outlined 
time frames. The Sponsor will provide a list of project contacts for SAE receipt, telephone 
numbers, and mailing addresses 
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. 
7.7.3 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 (Woman of Child-Bearing Potential) enrolled in the study who becomes 
pregnant during the study or within 30 days after EOS should be followed through 
delivery or termination of the pregnancy. The Investigator will collect pregnancy 
information and report to the Sponsor within 24 hours of becoming aware of a 
participant’s pregnancy. Follow-up will generally not be required for longer than 6 to 8 
weeks beyond the estimated delivery date. Once pregnancy is confirmed, pregnant female 
participants will be immediately withdrawn from the study as outlined in Section 4.3.5.   
Where possible, the Investigator will also attempt to collect and report information on 
pregnancy outcomes of female partners of any male participants who were administered 
IMP in this study, if the female partner became pregnant during the study or within 90 
days after EOS. Appropriate signed informed consent will be required directly from the 
pregnant female partner to obtain and report this information. Any participant’s female 
partner who becomes pregnant during the study should be followed through delivery or 
termination of the pregnancy.  
7.8 
SAFETY REVIEW COMMITTEE (SRC)  
The Principal Investigator, Sponsor Medical Director and study Medical Monitor (at a 
minimum) will review the sentinel group (Group 1a) safety information up to and 
including Day 8 before deciding if the remainder of the cohort (Group 1b) should proceed.  


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8 STATISTICAL ANALYSES 
8.1 
GENERAL APPROACH 
The following sections describe the statistical analysis as it is foreseen during the planning 
phase of trial. A detailed Statistical Analysis Plan (SAP) will be finalised and approved 
prior to database lock and will provide details of the safety 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. Additional analytical plans will be prepared to detail the PK and PD analyses.  
Statistical analysis will be performed using SAS® software version 9.4 (or higher) (SAS 
Institute Inc., Cary, NC, USA).  
The general analytical approach for all safety endpoints will be descriptive in nature. 
Unless otherwise stated, the following statistical approaches will be taken: 
Continuous variables:   
Descriptive statistics will include the number of 
non-missing values, mean, standard deviation (SD), 
median, minimum, and maximum.  The minimum and 
maximum values will be presented to the same number of 
decimal places as recorded in the raw data; mean, median 
and SD will be presented to one more decimal place than 
the raw data. 
Categorical variables:   
Descriptive statistics will include frequency counts and 
percentages per 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. 
Imputation:   
No missing data will be imputed. 
Confidence intervals (CIs):  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. 
 
Baseline:   
Baseline will be defined as the last available assessment 
prior to the first IMP administration.  
 
Unscheduled assessments: 
Unscheduled visits will be excluded from visit-based 
summary tables. 
 
Early termination visits: 
Assessments conducted at Early Termination will be 
excluded from visit-based summary tables. 
8.2 
SAMPLE SIZE 
Eight healthy males or females, aged between 18 and 55 years old, who meet all of the 
inclusion criteria and none of the exclusion criteria will be enrolled in two sequential 
groups of 2 participants (Group 1a) and 6 participants (Group 1b).  
The 2 participants in Group 1a will serve as sentinels for Group 1b.  


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As the combination of Rux+AL has not been tested previously, a first-in-human approach 
of a sample size of 8 participants has been selected.  
8.3 
ANALYSIS SETS AND SUB-SETS 
8.3.1 Analysis Sets 
In the first instance, four (4) analysis datasets will be used for study analyses: Full 
Analysis Set (FAS), Safety Set, PK Set, and pSTAT3 Set. 
Additional analysis populations may be defined in the SAP. 
The number of participants in each analysis set will be summarised, with a corresponding 
listing. 
8.3.1.1 
Full Analysis Set 
The Full Analysis Set (FAS) will consist of all enrolled participants.  The FAS will be 
used to assess all participant disposition, baseline, demographic, treatment exposure and 
protocol deviation data. 
8.3.1.2 
Safety Set 
The Safety Set will include all enrolled participants who received at least one dose (full 
or partial) of IMP. The Safety Set will be used to assess all safety data.  
8.3.1.3 
PK Set 
The PK Set will include all enrolled participants who received at least one dose AL, Rux, 
and/or placebo and have sufficient samples for analysis. The PK Set will be used to assess 
all PK data. 
8.3.1.4 
pSTAT3 Set 
The pSTAT3 Set will include all enrolled participants who received at least one dose (full 
or partial) of IMP, a valid baseline pSTAT3 result and at least one valid post-baseline 
pSTAT3 result. The pSTAT3 Set will be used to assess all PD data.  
8.3.2 Sub-Sets 
Participants will be analysed in the following treatment groups:  
• AL+Rux (n=6; 1 participant from Group 1a + 5 participants from Group 1b) 
• AL+placebo (n=2; 1 participant from Group 1a and 1 participant from Group 1b) 
Unless otherwise stated, summary tables will be presented by treatment group and overall. 
8.4 
PARTICIPANT DISPOSITION 
A listing of participant disposition will present participant dates of informed consent, 
randomisation, key visits as well as study completion details. Early termination data, 
including the reason for early termination will be listed in an additional listing.  
A participant disposition summary table will present, at a minimum, the number of 
participants who completed the study per protocol and the number of participants who 
discontinued classified by reason for early termination.   


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8.5 
DEMOGRAPHICS AND BASELINE CHARACTERISTICS 
All descriptive statistics for demographic and baseline characteristic parameters will be 
evaluated using the Full Analysis Set.  
8.5.1 Demographics 
Demographic data will be listed for all enrolled participants and summarised by treatment 
arm and overall.  
8.5.2 Medical History 
All medical history data will be listed, grouped by participant. 
Medical history will be coded using the Medical Dictionary for Regulatory Activities 
(MedRA) and summarised by system organ class (SOC) and preferred term (PT).  
8.5.3 Prior Medications 
Prior medications will be listed for all enrolled participants. Prior medications are defined 
as any medication that is started before administration of IMP, regardless of when it 
ended. If a medication has a missing or partial missing start/end date or time and it cannot 
be determined whether it was taken before IMP or concomitantly, it will be considered as 
prior and concomitant. 
8.5.4 Eligibility 
Eligibility will be listed for all enrolled participants, including findings for drug and 
alcohol screening.  
8.6 
PROTOCOL DEVIATIONS 
All protocol deviations will be listed. A protocol deviation summary table will present the 
total number of protocol deviations as well as the number of participants who reported at 
least one protocol deviation, broken down by deviation type including (but not limited 
to): 
• Informed consent 
• Eligibility 
• Visit not done 
• Visit performed out of window 
• Study procedure not done 
• Study procedure done out of window 
• Safety reporting 
• Investigational Product 
• Privacy and Data Protection 
• Concomitant Medication 
• Other 
8.7 
TREATMENT EXPOSURE 
Participant exposure to all protocol-specified treatments will be listed and summarised.   


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8.8 
SAFETY (PRIMARY ENDPOINT) 
8.8.1 Safety Endpoints 
Safety and tolerability will be assessed by clinical review of the following parameters:  
• AEs (including SAEs and AESIs) 
• Vital signs and other body measurements including respiratory rate and body 
temperature 
• 12-lead ECG 
• Haematology, chemistry, coagulation and urinalysis 
• Physical examination 
All descriptive statistics for safety parameters will be evaluated using the Safety Set.    
8.8.2 Adverse Events  
All AE data will be listed for each participant, including severity, relationship to IMPs, 
relationship to non-IMP protocol-specific treatments, outcome and actions taken. In 
addition, listings of AEs leading to discontinuation of the study, SAEs and deaths, will be 
provided as applicable.  
All AE summaries will be restricted to TEAEs only, where a TEAE is defined as an AE 
that commences on, or after, the first administration of IMP. 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 started and/or stopped prior to the first 
administration of IMP.  
All reported TEAEs, including SAEs and AESIs, will be mapped to standard MedDRA 
coding terms and grouped by SOC and PT.   
At a minimum, the following AE summary tables will be provided: 
• Overall summary of TEAEs 
• All TEAEs 
• TEAEs by severity 
• TEAEs by relationship (separate tables for each relationship type assessed) 
• Serious TEAEs  
• TEAEs leading to study withdrawal  
• TEAEs of Special Interest (AESIs) 
For the summaries of TEAEs, participants who experience the same AE (in terms of the 
MedDRA preferred term) more than once will only be counted once.  
8.8.3 Concomitant Medications 
Medications used in this study will be coded using the World Health Organisation Drug 
Dictionary Enhanced (WHO-DDE) and/or the World Health Organisation Drug Global 
(WHODrug Global). 
Concomitant medications are defined as medications continued or newly received at or 
after administration of IMP, through to the End of Study visit. 
If a medication has a missing or partial missing start/end date or time and it cannot be 
determined whether it was taken before initial treatment or concomitantly, it will be 
considered as prior and concomitant. 


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Concomitant medications will be summarised by Anatomical Therapeutic Class (ATC) 
and preferred name (PN). The summary tables will show the number and percentage of 
participants taking each medication by ATC and PN. 
Participants who take the same medication (in terms of the ATC and PN) more than once 
will only be counted once for that medication. 
8.8.4 Laboratory Findings 
Laboratory parameters will be listed by participant and visit, including: 
• haematology, 
• biochemistry, 
• coagulation, 
• urinalysis, 
• serology, 
• FSH, and 
• pregnancy test. 
All laboratory findings will be listed with flags for values outside the laboratory normal 
reference ranges and clinical significance status. 
Haematology, biochemistry and continuous urinalysis laboratory data will be summarised 
for each scheduled visit, including observed values and absolute change from each 
baseline. Categorical urinalysis results will be summarised for each scheduled visit using 
frequency tabulations. 
Any available microscopic urinalysis will be listed. 
8.8.5 Physical Examination 
Physical examination parameters will be listed for all participants and visits. 
8.8.6 Vital Signs and other Body Measurements.  
Vital sign parameters and other body measurements will be listed for all participants and 
visits, with three separate listings:  
• Vital signs 
• Respiratory rate and temperature 
• Height and weight 
Observed values, as well as absolute changes from each baseline, will be summarised 
descriptively vital signs, respiratory rate and temperature by scheduled visit, with two 
separate tables: 
• Vital signs 
• Respiratory rate and temperature 
8.8.7 12-Lead ECG 
ECG parameters will be listed by participants and visits.  
Observed values, as well as absolute changes from each baseline, will be summarised 
descriptively for all ECG parameters by visit.  
Safety analysis of ECGs will include in particular the number of participants during the 
study with: 
•  QTcF and/or QTcB prolongation of more than 30 msec and 60 msec; and/or  


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• QTcF and/or QTcB > 450 msec.  
8.8.8 Beck Depression Inventory 
Beck Depression Inventory (BDI-II) data will be listed for all participants. 
8.9 
PHARMACOKINETIC ENDPOINTS 
8.9.1 Pharmacokinetic parameters 
Secondary endpoint PK parameters of artemether, DHA, lumefantrine, and Rux will be 
estimated using non-compartmental methods from plasma concentration-time data:  
• AUClast 
• AUC0-  
• AUC0-8  
• AUC60-72 
• Cmax (first and last dose) 
• tmax (first and last dose) 
• Elimination t½ 
• tlag 
• C168h (for lumefantrine only) 
• CL/F 
• Vz/F 
• λz 
For calculation of descriptive statistics of plasma concentrations, values below the lower 
limit of quantitation will be set to zero.  
PK parameters will be determined using STATA® (version 14.0 or higher). 
8.10 PHARMACODYNAMIC ENDPOINTS 
8.10.1 pSTAT3 levels 
The secondary endpoint pSTAT3 inhibition ex-vivo on whole blood cells will be assessed 
using a validated ELISA. pSTAT3 inhibition will be expressed as a percentage and 
summarised by percentage inhibition pre and post IMP administration and as change in 
percentage between pre and post IMP administration. The following analyses will be 
conducted: 
• Within-participant comparison of each participant before AL+Rux or AL+placebo 
administration, and 12 hours after first administration of AL.  
• Between-participants comparison of participants who receive AL+Rux and 
participants who receive AL+placebo 
8.11 INTERIM ANALYSES 
This trial has no formal interim analyses other than review of safety and tolerability as 
described in Section 7.8.  


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9 STUDY ADMINISTRATION 
9.1 
ETHICAL CONSIDERATIONS 
9.1.1 Ethical Principles 
This clinical study was designed and shall be implemented and reported in accordance 
with the Declaration of Helsinki, the Integrated Addendum to ICH E6(R1): Guideline for 
Good Clinical Practice E6(R2) dated 09 November 2016 and the National Statement on 
Ethical Conduct in Human Research, (2007 – updated 2018). 
9.1.2 Informed Consent 
Eligible participants may only be included in the study after providing written (witnessed, 
where required by law or regulation), IRB/IEC-approved informed consent. Informed 
consent must be obtained before conducting any study-specific procedures (i.e. all of the 
procedures described in the protocol). The process of obtaining informed consent should 
be documented in the participant source documents and a copy of the signed patient 
information and consent form provided to the participant. 
9.1.3 Investigator 
and 
Human 
Research 
Ethics 
Committee 
(HREC) 
Responsibilities  
Before initiating a trial, the investigator/institution should obtain approval/favorable 
opinion from the Human Research Ethics Committee (HREC) for the trial protocol, 
written informed consent form, consent form updates, participant recruitment procedures 
(e.g., advertisements) and any other written information to be provided to participants.  
Prior to study start, the investigator is required to sign a protocol signature page 
confirming his/her agreement to conduct the study in accordance with these documents 
and all of the instructions and procedures found in this protocol and to give access to all 
relevant data and records to the Sponsor appointed monitors, auditors, Quality Assurance 
representatives, HREC representatives, and regulatory authorities as required. If an 
inspection of the clinical site is requested by a regulatory authority, the investigator must 
inform the Sponsor immediately that this request has been made. 
9.2 
PROTOCOL ADHERENCE 
This protocol defines the study objectives, the study procedures and the data to be 
collected on study participants. Additional assessments required to ensure safety of 
participants should be administered as deemed necessary on a case by case basis. In this 
instance the Sponsor medical monitor must be advised before or as soon as possible after 
such assessments are conducted.   
Under no circumstances should an investigator collect additional data or conduct any 
additional procedures for any research related purpose involving any investigational 
drugs. Investigators ascertain they will apply due diligence to avoid protocol deviations. 
If an investigator feels a protocol deviation would improve the conduct of the study this 
must be considered a protocol amendment, and unless such an amendment is agreed upon 
by the Sponsor and approved by the HREC it cannot be implemented. All significant 
protocol deviations will be recorded and reported in the CSR. 
9.3 
PROTOCOL AMENDMENTS 
Any change or addition to the protocol can only be made in a written protocol amendment 
that must be approved by the Sponsor and the HREC prior to implementation. Only 
amendments that are intended to eliminate an apparent immediate hazard to participants 


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may be implemented immediately provided the Sponsors medical monitor is notified as 
soon as possible after the event and the reviewing HREC is subsequently notified. 
Notwithstanding the need for approval of formal protocol amendments, the investigator 
is expected to take any immediate action required for the safety of any participant included 
in this study, even if this action represents a deviation from the protocol. In such cases, 
the Medical Monitor and Sponsor must be notified immediately. 
9.4 
DATA HANDLING AND RECORD KEEPING 
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 Principal Investigator 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.   
Study-related participant data will be entered into electronic case report forms (CRFs), 
except for data that may be transmitted to the Sponsor or CRO electronically (such as 
laboratory data). The Principal Investigator is responsible for ensuring that accurate 
source documents for all data entered into the CRF are maintained at the study site.  
All study documents including source documents and signed PICFs must be retained by 
the Principal Investigator for at least 15 years and according to local regulatory 
requirements. No study documents may be destroyed or transferred during the retention 
period without the Sponsor being directly notified in writing. 
9.5 
QUALITY CONTROL AND QUALITY ASSURANCE 
The Sponsor or CRO maintains responsibility for quality assurance of the data and for 
data management, and retains accountability for actions delegated to other parties 
(including CRO). Study monitors appointed by the Sponsor or CRO will conduct ongoing 
visits to the study sites (and/or remote monitoring according to Sponsor or CRO standard 
operating procedures (SOPs) if required due to COVID-19 restrictions) to confirm the 
CRF data is accurate according to source documents and complete, and that the study is 
being appropriately conducted according to the protocol, ICH GCP and local regulatory 
requirements. Details may also be provided in the Monitoring Plan.  
9.6 
PUBLICATION POLICY 
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. 
Results of this trial will be either submitted for publication and/or posted in a publicly 
accessible database of clinical trial results.   
 
 


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10 REFERENCES 
1. World 
Health 
Organization 
(WHO), 
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malaria 
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2019. 
https://www.who.int/publications-detail/world-malaria-report-2019, 
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January 2020. 
2. Cohen S, Mc Gregor I, Carrington S. Gamma-globulin and acquired immunity to 
human malaria. Nature. 1961;192:733-7. 
3. Stanisic DI, Fowkes FJ, Koinari M, Javati S, Lin E, Kiniboro B, et al. Acquisition of 
antibodies against Plasmodium falciparum merozoites and malaria immunity in young 
children and the influence of age, force of infection, and magnitude of response. Infect 
Immun. 2015;83(2):646-60. 
4. Teo A, Feng G, Brown GV, Beeson JG, Rogerson SJ. Functional antibodies and 
protection against blood-stage malaria. Trends in Parasitology. 2016;32(11):887-98. 
5. Stephens R, Albano FR, Quin S, Pascal BJ, Harrison V, Stockinger B, et al. Malaria-
specific transgenic CD4+ T cells protect immunodeficient mice from lethal infection 
and demonstrate requirement for a protective threshold of antibody production for 
parasite clearance. Blood. 2005;106(5):1676. 
6. Zander RA, Vijay R, Pack AD, Guthmiller JJ, Graham AC, Lindner SE, et al. Th1-
like plasmodium-specific memory CD4+ T cells support humoral immunity. Cell Rep. 
2017;21(7):1839-52. 
7. Figueiredo MM, Costa PAC, Diniz SQ, Henriques PM, Kano FS, Tada MS, et al. T 
follicular helper cells regulate the activation of B lymphocytes and antibody 
production during Plasmodium vivax infection. PLoS Pathog. 2017;13(7):e1006484. 
8. MacLeod MKL, David A, McKee AS, Crawford F, Kappler JW, Marrack P. Memory 
CD4 T cells that express CXCR5 provide accelerated help to B cells. J Immunol. 
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9. Su Z, Stevenson MM. Central role of endogenous gamma interferon in protective 
immunity against blood-stage Plasmodium chabaudi AS infection. Infect Immun. 
2000 Aug;68(8):4399-406. 
10. Su Z, Fortin A, Gros P, Stevenson MM. Opsonin-independent phagocytosis: an 
effector mechanism against acute blood-stage Plasmodium chabaudi AS infection. J 
Infect Dis. 2002 Nov 1;186(9):1321-9. 
11. Haque A, Best SE, Amante FH, Mustafah S, Desbarrieres L, de Labastida F, et al. 
CD4+ natural regulatory T cells prevent experimental cerebral malaria via CTLA-4 
when expanded in vivo. PLoS Pathog. 2010 Dec 9;6(12):e1001221. 
12. Sun T, Holowka T, Song Y, Zierow S, Leng L, Chen Y, et al. A Plasmodium-encoded 
cytokine suppresses T-cell immunity during malaria. Proc Natl Acad Sci U S A. 
2012;109(31):E2117-26. 
13. Xu H, Wipasa J, Yan H, Zeng M, Makobongo MO, Finkelman FD, et al. The 
mechanism and significance of deletion of parasite-specific CD4(+) T cells in malaria 
infection. J Exp Med. 2002;195(7):881-92. 
14. Costa PA, Leoratti FM, Figueiredo MM, Tada MS, Pereira DB, Junqueira C, et al. 
Induction of inhibitory receptors on T cells during Plasmodium vivax malaria impairs 
cytokine production. J Infect Dis. 2015;212(12):1999-2010. 


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15. Illingworth J, Butler NS, Roetynck S, Mwacharo J, Pierce SK, Bejon P, et al. Chronic 
exposure to Plasmodium falciparum is associated with phenotypic evidence of B and 
T cell exhaustion. J Immunol. 2013 Feb 1;190(3):1038-47. 
16. Jagannathan P, Eccles-James I, Bowen K, Nankya F, Auma A, Wamala S, et al. 
IFNgamma/IL-10 co-producing cells dominate the CD4 response to malaria in highly 
exposed children. PLoS Pathog. 2014 Jan;10(1):e1003864. 
17. Jankovic D, Kullberg MC, Feng CG, Goldszmid RS, Collazo CM, Wilson M, et al. 
Conventional T-bet(+)Foxp3(-) Th1 cells are the major source of host-protective 
regulatory IL-10 during intracellular protozoan infection. J Exp Med. 2007 Feb 
19;204(2):273-83. 
18. Obeng-Adjei N, Portugal S, Tran TM, Yazew TB, Skinner J, Li S, et al. Circulating 
Th1 cell-type Tfh cells that exhibit impaired B cell help are preferentially activated 
during acute malaria in children. Cell Rep. 2015;13(2):425-39. 
19. Portugal S, Tipton CM, Sohn H, Kone Y, Wang J, Li S, et al. Malaria-associated 
atypical memory B cells exhibit markedly reduced B cell receptor signaling and 
effector function. eLife. 2015;4:e07218. 
20. Ivashkiv LB, Donlin LT. Regulation of type I interferon responses. Nature reviews 
Immunology. 2014;14(1):36. 
21. Pestka S, Krause CD, Walter MR. Interferons, interferon‐like cytokines, and their 
receptors. Immunol Rev. 2004;202(1):8-32. 
22. McNab F, Mayer-Barber K, Sher A, Wack A, O'garra A. Type I interferons in 
infectious disease. Nat Rev Immunol. 2015;15(2):87. 
23. Aucan C, Walley A, Hennig B, Fitness J, Frodsham A, Zhang L, et al. Interferon-alpha 
receptor-1 (IFNAR1) variants are associated with protection against cerebral malaria 
in the Gambia. Genes Immun. 2003;4(4):275. 
24. Khor CC, Vannberg FO, Chapman SJ, Walley A, Aucan C, Loke H, et al. Positive 
replication and linkage disequilibrium mapping of the chromosome 21q22. 1 malaria 
susceptibility locus. Genes Immun. 2007;8(7):570. 
25. Lovegrove FE, Gharib SA, Patel SN, Hawkes CA, Kain KC, Liles WC. Expression 
microarray analysis implicates apoptosis and interferon-responsive mechanisms in 
susceptibility to experimental cerebral malaria. Am J Pathol. 2007;171(6):1894-903. 
26. Haque A, Best SE, Ammerdorffer A, Desbarrieres L, de Oca MM, Amante FH, et al. 
Type I interferons suppress CD4+ T‐cell‐dependent parasite control during blood‐
stage Plasmodium infection. Eur J Immunol. 2011;41(9):2688-98. 
27. Haque A, Best SE, De Oca MM, James KR, Ammerdorffer A, Edwards CL, et al. 
Type I IFN signaling in CD8–DCs impairs Th1-dependent malaria immunity. The 
Journal of clinical investigation. 2014;124(6):2483-96. 
28. Zander RA, Guthmiller JJ, Graham AC, Pope RL, Burke BE, Carr DJ, et al. Type I 
interferons induce T regulatory 1 responses and restrict humoral immunity during 
experimental malaria. PLoS Pathog. 2016;12(10):e1005945. 
29. Montes de Oca M, Good MF, McCarthy JS, Engwerda CR. The impact of established 
immunoregulatory networks on vaccine efficacy and the development of immunity to 
malaria. J Immunol. 2016 Dec 15;197(12):4518-26. 


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30. Ladislau L, Suarez-Calvet X, Toquet S, Landon-Cardinal O, Amelin D, Depp M, et 
al. JAK inhibitor improves type I interferon induced damage: proof of concept in 
dermatomyositis. Brain. 2018;141(6):1609-21. 
31. Villarino AV, Kanno Y & O’Shea JJ. Mechanisms and consequences of Jak-STAT 
signaling in the immune system. Nat. Immunol. 2017(18):374-384. 
32. Shi JG, Chen X, McGee RF, Landman RR, Emm T, Lo Y, et al. The pharmacokinetics, 
pharmacodynamics, and safety of orally dosed INCB018424 phosphate in healthy 
volunteers. J Clin Pharmacol. 2011 Dec;51(12):1644-54. 
33. Harrison C, Kiladjian JJ, Al-Ali HK, Gisslinger H, Waltzman R, Stalbovskaya V, et 
al. JAK inhibition with ruxolitinib versus best available therapy for myelofibrosis. N 
Engl J Med. 2012;366(9):787-98. 
34. Verstovsek S, Mesa RA, Gotlib J, Levy RS, Gupta V, DiPersio JF, et al. A double-
blind, placebo-controlled trial of ruxolitinib for myelofibrosis. N Engl J Med. 
2012;366(9):799-807. 
35. Fremond ML, Rodero MP, Jeremiah N, Belot A, Jeziorski E, Duffy D, et al. Efficacy 
of the Janus kinase 1/2 inhibitor ruxolitinib in the treatment of vasculopathy associated 
with TMEM173-activating mutations in 3 children. J Allergy Clin Immunol. 
2016;138(6):1752-5.  
36. Medicines for Malaria Venture (MMV) Internal Communication, May 2020.  
37. Ogama Y, Mineyama T, Yamamoto A, Woo M, Shimada N, Amagasaki T, et al. A 
randomized dose-escalation study to assess the safety, tolerability, and 
pharmacokinetics of ruxolitinib (INC424) in healthy Japanese volunteers. Int J 
Hematol. 2013 Mar;97(3):351-9.  
38. Beck AT, Ward CH, Mendelson M, Mock J & Erbaugh J (1961). An Inventory of 
Measuring Depression. Archives of General Psychiatry. 4(6):561-571.  
39. Beck A T, Steer R A, Ball R & Ranieri W (1996). Comparison of Beck depression 
inventories – IA and II in psychiatric outpatients. Journal of personality assessment 
67(3): 588 – 597. 
 
 
 


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11 APPENDICES 
APPENDIX 1: CLINICALLY ACCEPTABLE RANGES FOR CLINICALLY 
IMPORTANT STUDY INCLUSION/EXCLUSION LABORATORY TESTS 
V2.0 16 JULY 2020 
 
Test 
Acceptable Inclusion Range 
Low 
High 
Sodium 
130 mmol/L  
150 mmol/L  
Potassium 
3.0 mmol/L  
5.5 mmol/L  
Chloride 
0.95 x LLN  
1.05 x ULN   
Magnesium  
0.95 x LLN  
1.05 x ULN   
Calcium (Corrected) 
0.95 x LLN  
1.05 x ULN  
Phosphate  
0.95 x LLN  
1.05 x ULN  
Glucose Fasted 
N/A 
1.0 x ULN  
Urea 
N/A 
1.75 x ULN 
Creatinine 
N/A 
1.0 x ULN  
eGFR (CKD-EPI equation) 
> 59 ml/min/1.73m2  
N/A  
Creatine kinase 
N/A 
< 2 x ULN 
Uric acid  
N/A  
1.1 x ULN  
Total Protein 
≥ 0.85 x LLN  
≤ 1.25 x ULN  
Albumin 
≥ 0.85 x LLN 
≤ 1.25 x ULN 
Total Bilirubin 
N/A 
1.25 x ULN 
Direct Bilirubin 
N/A 
1.25 x ULN 
ALP 
N/A  
1.5 x ULN 
AST 
N/A 
1 x ULN 
ALT 
N/A 
1 x ULN 
GGT 
N/A 
1.5 x ULN 


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Test 
Acceptable Inclusion Range 
Low 
High 
Lactate Dehydrogenase 
0.9 x LLN 
1.1 x ULN 
Total cholesterol 
N/A 
1.2 x ULN  
HDL  
> 1.1 mmol/L  
N/A  
LDL  
N/A  
< 3.34 mmol/L  
Triglycerides  
NA  
1.2 x ULN  
Prothrombin time INR and 
APTT 
1.0 x LLN  
1.0 x ULN  
Haemoglobin 
0.9 x LLN  
1.1.x ULN 
Red blood count  
0.9 x LLN 
1.1 x ULN  
Reticulocytes  
0.9 x LLN  
1.1 x ULN  
MCV  
0.9 x LLN  
1.1 x ULN  
Platelets  
> 200 x109/L  
1.1 x ULN  
White Blood Cells 
0.9 x LLN  
1.1 x ULN  
Neutrophils  
1.0 x LLN  
1.0x ULN  
Lymphocytes 
1.0 x LLN  
1.0 x ULN  
Monocytes 
N/A 
1.2 x ULN  
Eosinophils 
N/A 
1.0 x ULN  
Basophils 
N/A 
2.0 x ULN  
Urinalysis  
Glucose  
(quantitative)  
N/A  
≤ 1.9 mmol/L 
Protein  
(quantitative) 
N/A 
< 1+ 
Red Blood Cells (quantitative) N/A 
<10* x 106/L  
White Blood Cells 
(quantitative) 
N/A 
<10 x 106/L  
Bacteria  (any analysis) 
N/A  
< 1+  
Ketones (dipstick) 
N/A 
< 1+ 


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Test 
Acceptable Inclusion Range 
Low 
High 
Nitrate  (dipstick) 
N/A  
Negative    
Urobilinogen (dipstick) 
N/A 
< 1+ 
Bilirubin (dipstick) 
N/A 
< 1+ 
ABBREVIATIONS: ALP = alkaline phosphatase; ALT = alanine aminotransferase; APTT = 
activated partial thromboplastin time; AST = aspartate aminotransferase; CKD-EPI = Chronic 
Kidney Disease Epidemiology collaboration; eGFR = estimated glomerular filtration rate; GGT 
= gamma-glutamyl transferase; HDL = high density lipoprotein; INR = international normalised 
ratio; LDL = low density lipoprotein; LLN = lower limit of normal; MCV = mean corpuscular 
volume; N/A = not applicable; ULN = upper limit of normal.  
*A result ≥ 10 x 106/L is acceptable for female participants currently menstruating. 
   
 
 
 
 

SAP_001.pdf: 
Protocol Number: MMV_Ruxolitinib_19_01  
16 December 2020 
Document status: SAP Version 1.0 (FINAL) 
BM S01-A, v2.0 
Statistical Analysis Plan 
Template Effective: 18/JAN/2018 
CONFIDENTIAL 
Page 1 of 22 
 
 
 
STATISTICAL ANALYSIS PLAN 
A randomised, single blind, placebo controlled, phase 1 trial to evaluate the safety, 
tolerability, pharmacokinetic and pharmacodynamic activity of Ruxolitinib when 
co-administered with artemether-lumefantrine in healthy participants 
Protocol No.: MMV_Ruxolitinib_19_01 
Product Codes: Ruxolitinib Phosphate (Jakavi®), Artemether-lumefantrine (Riamet®) 
 
PREPARED FOR: 
 
Medicines for Malaria Venture (MMV) 
20, Route de Pré-Bois 
1215 Geneva 15 
Switzerland 
 
PREPARED BY: 
 
Southern Star Research 
Level 1, 1 Merriwa St 
Gordon, NSW, 2072  
Australia  
 
DATE OF ISSUE: 
 
16 December 2020  
 
VERSION/STATUS: 
 
Version 1.0 (FINAL) 
 
AUTHOR: 
 
Dr Elisa Young 
 
 
 




Protocol Number: MMV_Ruxolitinib_19_01  
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TABLE OF CONTENTS 
 
1 
INTRODUCTION ............................................................................................................................... 5 
2 
PROJECT OVERVIEW ....................................................................................................................... 5 
2.1 
Study Design ......................................................................................................................................... 5 
2.2 
Sample Size ........................................................................................................................................... 7 
2.3 
Treatment Assignment and Randomization ......................................................................................... 7 
2.4 
Study Objectives and Endpoints ........................................................................................................... 8 
3 
STATISTICAL CONSIDERATIONS ....................................................................................................... 9 
3.1 
Data Capture ....................................................................................................................................... 10 
3.2 
Statistical Programming...................................................................................................................... 11 
4 
ANALYSIS SETS .............................................................................................................................. 11 
4.1 
Analysis Set Descriptions .................................................................................................................... 11 
5 
PARTICIPANT DISPOSITION ........................................................................................................... 12 
5.1 
Disposition .......................................................................................................................................... 12 
5.2 
Clinical Trial Unit Admission, Confinement and Discharge ................................................................ 12 
6 
PROTOCOL DEVIATIONS ................................................................................................................ 13 
7 
DEMOGRAPHIC AND BASELINE CHARACTERISTICS ...................................................................... 13 
7.1 
Demographics ..................................................................................................................................... 13 
7.2 
Medical history ................................................................................................................................... 14 
7.3 
Prior Medications ............................................................................................................................... 14 
7.4 
Body Measurements .......................................................................................................................... 14 
7.5 
Eligibility ............................................................................................................................................. 14 
7.6 
Drug Screen ........................................................................................................................................ 14 
8 
TREATMENT EXPOSURE ................................................................................................................ 14 
9 
PHARMACOKINETICS .................................................................................................................... 15 
10 
PHARMACODYNAMICS.............................................................................................................. 15 
11 
SAFETY ....................................................................................................................................... 16 
11.1 
Safety Endpoints ................................................................................................................................. 16 
11.2 
Adverse Events ................................................................................................................................... 16 
11.3 
Concomitant Medications .................................................................................................................. 18 
11.4 
Laboratory .......................................................................................................................................... 18 
11.5 
Pregnancy Test and FSH ..................................................................................................................... 19 
11.6 
Vital Signs............................................................................................................................................ 20 
11.7 
Physical Examination .......................................................................................................................... 20 
11.8 
12-Lead ECG ........................................................................................................................................ 21 
11.9 
Beck Depression Inventory ................................................................................................................. 21 
12 
HANDLING OF MISSING DATA ................................................................................................... 22 
13 
CHANGES AND CLARIFICATIONS TO THE PLANNED ANALYSIS .................................................. 22 
14 
INTERIM ANALYSES ................................................................................................................... 22 
15 
SOFTWARE ................................................................................................................................ 22 
16 
REFERENCES .............................................................................................................................. 22 
 
 
 
 


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ABBREVIATIONS 
 
ADaM 
Analysis Data Model 
AE 
Adverse Event 
AESI 
Adverse Event of Special Interest 
AL 
Artemether-Lumefantrine 
ALP 
Alkaline phosphatase 
ALT 
Alanine Aminotransferase 
APTT 
Activated partial thromboplastin time 
AST 
Aspartate Aminotransferase 
ATC 
Anatomical Therapeutic Chemical 
β-HCG 
β-Human Chorionic Gonadotropin 
BDI 
Beck Depression Inventory 
CDISC 
Clinical Data Interchange Standards Consortium 
CI 
Confidence Interval 
CK 
Creatine Kinase 
CRA 
Clinical Research Associate 
CSR 
Clinical Study Report 
CTCAE 
Common Terminology Criteria for Adverse Events 
DBP 
Diastolic Blood Pressure 
DHA 
Dihydroartemisinin 
DTS 
Data Transfer Specification 
ECG 
Electrocardiography 
eCRF 
Electronic Case Report Form 
EDC 
Electronic Data capture 
eGFR 
Estimated Glomerular Filtration Rate 
FAS 
Full Analysis Set 
FSH 
Follicular Stimulating Hormone 
GGT 
Gamma Glutamyl Transpeptidase 
HBsAg 
Hepatitis B Surface Antigen 
HDL 
High Density Lipoprotein 
HEENT 
Head, Eyes, Ears, Nose, Throat 
HIV 
Human Immunodefiniciency Virus 
LDH 
Lactate Dehydrogenase 
LDL 
Low Density Lipoprotein 
MCV 
Mean Cell Volume 
MedDRA 
Medical Dictionary for Regulatory Activities 
PD 
Pharmacodynamics 
PK 
Pharmacokinetics 
PN 
Preferred Name 
PT 
Preferred Term 
QTcB 
Corrected QT interval with Bazett’s Formula 
QTcF 
Corrected QT interval with Fridericia’s Formula 
RCC 
Red Cell Count 
RDW 
Red Cell Distribution Width 
RUX 
Ruxolitinib 
SAE 
Serious Adverse Event 
SAP 
Statistical Analysis Plan 
SAS 
Statistical Analysis System 
SBP 
Systolic Blood Pressure 
SBQ 
Swiss BioQuant Central 
SD 
Standard Deviation 
SDTM 
Study Data Tabulation Model 
SOC 
System Organ Class 
SOP 
Standard Operating Procedure 
SRC 
Safety Review Committee 
SSR 
Southern Star Research 
TEAE 
Treatment Emergent Adverse Event 
WCC 
White Cell Count 
WHODRL 
World Health Organization-Drug Reference List 
 


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1 
INTRODUCTION 
This Statistical Analysis Plan (SAP) provides the outline for the statistical analysis deliverables 
required for the MMV_Ruxolitinib_19_01 study. 
The planned analyses identified in this SAP may be included in clinical study reports (CSRs), 
regulatory submissions, or future manuscripts. In addition, post hoc exploratory analyses not 
necessarily identified in this SAP may be performed to further examine study data.  Any post hoc, or 
unplanned, exploratory analyses performed will be clearly identified as such in the final CSR. 
Any significant changes from planned analyses will also be described in the final CSR. 
A separate pharmacokinetic (PK) analysis plan will be prepared to detail the PK related analyses. 
2 
PROJECT OVERVIEW 
2.1 
Study Design 
This  Phase1 study intends to evaluate whether the JAK1 inhibitor ruxolitinib (RUX) could be safely 
co-administered with the registered antimalarial (artemether-lumefantrine/AL), for potential use in 
the future as an “immune booster” in patients treated for an acute episode of . Along with standard 
Phase1 objectives (safety, tolerability and PK), the study also aims to evaluate whether the 
pharmacodynamic activity of RUX (pSTAT3 inhibition) could be impacted in the presence of AL.  
This study is a randomised, single-blind, placebo-controlled, single centre phase 1 trial designed to 
assess the safety, tolerability, PK and pharmacodynamics (PD) of combined oral doses of:  
• 
artemether-lumefantrine (AL; 20 mg/120 mg) and Ruxolitinib (Rux; 20 mg), or  
• 
artemether-lumefantrine (AL; 20 mg/120 mg) and placebo. 
The study will be composed of 2 groups (Group 1a and Group 1b) to be enrolled sequentially. Group 
1a will participate first as the sentinel safety group.  
The study is designed specifically to monitor safety and tolerability of the combination of AL+Rux, 
and to obtain PK and preliminary PD data. Eight participants will be randomised to receive AL+Rux 
or AL+placebo in a 3:1 ratio overall (1:1 in Group 1a, and 5:1 in Group 1b). This is typical of 
first-in-human studies, and this approach is used in this study as the combination of AL+Rux has not 
been studied previously. 
The study schema is presented in Figure 1. 


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Figure 1: Study Schema 
 
 


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2.2 
Sample Size 
Eight healthy males or females, aged between 18 and 55 years old, who meet all of the inclusion 
criteria and none of the exclusion criteria will be enrolled in two sequential groups of 2 participants 
(Group 1a) and 6 participants (Group 1b).  
The 2 participants in Group 1a will serve as sentinels for Group 1b.  
As the combination of Rux+AL has not been tested previously, a first-in-human approach of a 
sample size of 8 participants has been selected.  
2.3 
Treatment Assignment and Randomization 
Randomisation procedures are described in the Randomisation Plan and Randomisation Procedure.  
A randomisation number will be allocated to each participant as per the randomisation schedule 
before the administration of IMPs (AL+Rux or AL+placebo).  
Overall, randomisation will be in a ration of 3:1 (active:placebo): 
• 
Participants in Group 1a will be randomised to a treatment allocation (AL+Rux or 
AL+placebo) in a ratio of 1:1. The randomisation will be single blinded (treatment known to 
Investigator but not participant). 
• 
Participants in Group 1b will be randomised to a treatment allocation (AL+Rux or 
AL+placebo) in a ratio of 5:1. The randomisation will be single blinded (treatment known to 
Investigator but not participant). 
2.3.1 Replacement of Withdrawn/Discontinued Participants 
• Participants who sign the informed consent form and are randomised but do not receive IMP may 
be replaced.  Replacement volunteers are permitted as back-up.  
• Participants who sign the informed consent form, and are randomised and receive at least one 
dose of IMP, and subsequently withdraw, or are withdrawn or discontinued from the trial, may only 
be replaced after mutual agreement between the Sponsor and the Principal Investigator or 
delegate. The decision regarding the replacement of participants will be documented. 
 
 


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2.4 
Study Objectives and Endpoints 
Primary Objective  
Primary Endpoints 
1. To assess the safety and 
tolerability of 3-day b.i.d dosing of 
AL+Rux and AL+placebo.  
1. Incidence, severity, and relationship of observed 
and self-reported adverse events (AEs) up to 28 
days 
after 
AL+Rux 
and 
AL+placebo 
administration in all participants by treatment 
regimen.  
2. Clinical 
laboratory 
evaluations 
including 
haematology, biochemistry, and urinalysis. 
3. Vital signs (blood pressure and heart rate) 
4. 12-lead standard ECG: QT, QTcB and QTcF; 
HR, PR, QRS. 
Secondary Objectives 
Secondary Endpoints 
1. To assess the effect of AL+Rux 
and AL+placebo on pSTAT3 
inhibition.  
1. pSTAT3 inhibition ex-vivo on whole blood cells 
by: 
• pSTAT3 levels pre- and post- AL+Rux 
administration intra participant and, 
• pSTAT3 levels per treatment regimen post 
AL+Rux and post AL+placebo administration  
2. To characterise the PK profile of 
artemether and its major metabolite 
dihydroartemisinin [DHA], 
lumefantrine, and Rux (part 1 and 2). 
2. PK parameters of artemether, DHA, 
lumefantrine, and Rux using non-compartmental 
methods: AUClast, AUC0-∞, AUC0-8, AUC60-72, Cmax 
(first and last dose), tmax (first and last dose), 
elimination half-life (t½), tlag, C168h (for lumefantrine 
only), CL/F, Vz/F and λz in all participants. 
3. To assess additional safety 
variables (tympanic body temperature 
and respiration rate) 
3. Safety variables tympanic body temperature 
(°Celcius) and respiratory rate. 
 
 
 


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3 
STATISTICAL CONSIDERATIONS 
Data analysis will be performed according to Southern Star Research’s Standard Operating 
Procedures (SOPs). 
The general analytical approach for all safety endpoints will be descriptive in nature. Unless 
otherwise stated, the following statistical approaches will be taken: 
Continuous variables:   
Descriptive statistics will include the number of non-missing 
values, mean, standard deviation (SD), median, minimum, and 
maximum.  The minimum and maximum values will be presented 
to the same number of decimal places as recorded in the raw data; 
mean, median and SD will be presented to one more decimal place 
than the raw data. 
Categorical variables:   
Descriptive 
statistics 
will 
include 
frequency 
counts 
and 
percentages per 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. 
Imputation:   
No missing data will be imputed. 
Confidence intervals (CIs):  
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. 
Unscheduled assessments:  
Unscheduled visits will be excluded from visit-based summary 
tables. 
Early termination visits:  
Assessments conducted at Early Termination will be excluded 
from visit-based summary tables. 
 
 


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3.1 
Data Capture 
3.1.1 Database 
The primary method of data collection is via the study database, developed within the chosen 
Electronic Data Capture (EDC) platform, IBM Clinical Development. The database has been 
designed based on the final protocol, the system/core configuration, electronic Case Report Form 
(eCRF) specifications and/or mock eCRF and consistency check specifications. 
Data will  be entered directly into the EDC system. Site-collected data will be entered directly from 
source notes at the site and will be verified by Clinical Research Associates (CRAs) to ensure data 
integrity.  
Refer to the Data Management Plan for further details. 
3.1.2 Third Party Data 
3.1.2.1 Safety Laboratory (SydPath) 
Central safety laboratory data will be received from SydPath as specified in the SydPath Data 
Transfer Specification (DTS).  An initial transfer will be delivered prior to database lock and 
reconciled against CRF data.  Following successful reconciliation and resolution of any data issues, 
the data will be incorporated into the End of Study analysis. 
No unit conversion of laboratory data will be performed. 
3.1.2.2 PK Laboratory (SBQ) 
PK samples will be analysed by Swiss BioQuant Central (SBQ).  Final PK assay data will be 
transferred to SSR, as specified in the SBQ DTS, for incorporation into the PC SDTM. 
3.1.2.3 PK  Analysis and Modelling 
PK assay results will be transferred from SBQ to Paul van Giersbergen for PK analysis and 
modelling. PK parameter data will be transferred to SSR, as specified in a DTS, for incorporation 
into the PP SDTM. A PK consultant (Paul van Giersbergen) will perform the pk analysis according to 
the attached plan (Appendix 1). 
3.1.2.4 Pharmacodynamics - pSTAT (TetraQ) 
pSTAT laboratory results will be received from TetraQ as specified in the TetraQ DTS.  An initial 
transfer will be delivered prior to database lock and reconciled against CRF data.  Following 
successful reconciliation and resolution of any data issues, a Tetra Q consultant (Stephanie Reuter 
Lange) will perform the pharmacodynamic analysis according to the attached plan (Appendix 2).  
 
 


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3.2 
Statistical Programming 
3.2.1 Programming Specifications 
A programming specifications document will be prepared to detail the SAS programming of CDISC 
(SDTM and ADaM) datasets and listings, tables and figures. 
3.2.2 Baseline 
Baseline will be defined as the last available assessment prior to the first IMP administration.  
3.2.3 Change from Baseline 
Change from Baseline will be calculated as: 
𝑐𝑐ℎ𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎 𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓 𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏= (𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝𝑝 𝑣𝑣𝑣𝑣𝑣𝑣𝑣𝑣𝑣𝑣) −𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏𝑏 𝑣𝑣𝑣𝑣𝑣𝑣𝑣𝑣𝑣𝑣 
3.2.4 Study Day 
Study Day will be derived as the number of days relative to date of first administration of AL, where 
the day of first administration = 1. 
3.2.5 Listings, Tables and Figures 
Listings, tables and figures will be delivered as individual .rtf files in accordance with the mock 
listings, tables and figures, with separate sets of outputs for each study part. 
3.2.6 Treatment Groups 
Participants will be analysed in the following treatment groups:  
• 
AL+Rux (n=6; 1 participant from Group 1a + 5 participants from Group 1b) 
• 
AL+placebo (n=2; 1 participant from Group 1a and 1 participant from Group 1b) 
 
Unless otherwise stated, summary tables will be presented by treatment group and overall. 
4 
ANALYSIS SETS 
In the first instance, four (4) analysis sets will be used for the analyses: Full Analysis Set (FAS), 
Safety Set, PK Set and Pharmacodynamics (PD) Set.   
The number of participants in each analysis set will be summarised, with a corresponding listing. 
4.1 
Analysis Set Descriptions 
4.1.1 Full Analysis Set 
The FAS will consist of all enrolled participants.  The FAS will be used to assess all participant 
disposition, baseline, demographic, treatment exposure and protocol deviation data. 
4.1.2 Safety Set 
The Safety Set will include all enrolled participants who received at least one dose (full or partial) of 
AL. The Safety Set will be used to assess all safety data.  
4.1.3 PK Set 
The PK Set will include all enrolled participants who received at least one dose AL, Rux, and/or 
placebo and have sufficient samples for analysis. The PK Set will be used to assess all PK data. 
 Protocol deviations will be reviewed and exclusion of subjects from the PK analysis set will be decided 
during final data review prior to database lock. 


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4.1.4 Pharmacodynamics (PD) Set 
The PD Set will include all enrolled participants who received at least one dose (full or partial) of AL 
and Ruxolitinib or AL and Placebo, a valid baseline pSTAT3 result and at least one valid post-
baseline pSTAT3 result. The pSTAT3 Set will be used to assess all PD data.  
Protocol deviations will be reviewed and exclusion of subjects from the PD analysis set will be  
decided during final data review prior to database lock. 
5 
PARTICIPANT DISPOSITION 
5.1 
Disposition 
Analysis Set: FAS 
A listing of participant disposition will present: 
• 
Date of informed consent 
• 
Date of randomisation 
• 
Date of first dose of AL administration 
• 
Did the participant complete the study? 
• 
Date of completion / early withdrawal 
• 
Primary reason for early withdrawal (including instances where early termination was related 
to COVID-19) 
Detailed early withdrawal information will also be listed in a separate listing. 
If there are any deaths reported, a separate death listing will be prepared. 
The number and percentage of participants entering and discontinuing the study will be summarised 
along with the reason for discontinuation.  The participant disposition summary table will include: 
• 
Number of participants randomised 
• 
Number of participants who received at least one dose of study medication 
• 
Number of participants who completed the full study 
• 
Number of participants withdrawn from the study early 
• 
Reason for early withdrawal 
5.2 
Clinical Trial Unit Admission, Confinement and Discharge 
All  admission, confinement and discharge data will be listed. 
 


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6 
PROTOCOL DEVIATIONS 
Analysis Set: FAS 
All protocol deviations will be listed. A protocol deviation summary table will present the total number 
of protocol deviations as well as the number of participants who reported at least one protocol 
deviation, broken down by deviation category(minor/major) 
6.1.1 Definition of variables 
• 
Date deviation detected 
• 
Date of deviation 
• 
Deviation category (to be categorised by the Sponsor prior to Database Lock) 
o Minor 
o Major 
• 
Deviation type 
o Informed consent 
o Eligibility 
o Visit not done 
o Visit performed out of window 
o Study procedure not done 
o Study procedure done out of window 
o Safety reporting 
o Investigational Product 
o Privacy and Data Protection 
o Concomitant Medication 
o Other 
• 
Deviation description 
7 
DEMOGRAPHIC AND BASELINE CHARACTERISTICS 
Analysis Set: FAS 
7.1 
Demographics 
Demographic data will be listed for all enrolled participants and summarised by treatment arm and 
overall.  Data includes: 
• 
Date of birth 
• 
Age 
• 
Sex 
• 
Race 
• 
Women of Child-bearing potential 
 
 


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7.2 
Medical history 
All medical history data will be listed, grouped by participant. 
Medical history will be coded using the Medical Dictionary for Regulatory Activities (MedRA) and 
summarised by system organ class (SOC) and preferred term (PT).  
7.3 
Prior Medications 
Prior medications will be coded using the World Health Organization-Drug Reference List 
(WHODRL), the version of which will be included in the footnote of the listing. 
Prior medications will be listed for all enrolled participants. Prior medications are defined as any 
medication that is started before administration of IMP, regardless of when it ended. If a medication 
has a missing or partial missing start/end date or time and it cannot be determined whether it was 
taken before IMP or concomitantly, it will be considered as prior and concomitant. 
7.4 
Body Measurements 
Height and weight data at Screening will be listed for all participants. 
7.5 
Eligibility 
Eligibility will be listed for all enrolled participants. 
7.6 
Drug Screen 
Drug screen data will be listed for all participants. 
8 
TREATMENT EXPOSURE 
Analysis Set: Safety Set 
8.1.1 Parameters 
• 
Date/time of administration 
• 
Number of tablets administered 
8.1.2 Biostatistical methods 
Participant exposure to all protocol-specified treatments will be listed and summarised.  Exposure 
will be summarised as: 
• 
AL 
o Total dose (mg) 
o Total number of tablets 
o Compliance* 
• 
Ruxolitinib 
o Total dose (mg) 
o Total number of tablets 
o Compliance* 
 
* Compliance, derived as: 
𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶𝐶 (%) = 
𝑇𝑇𝑇𝑇𝑇𝑇𝑇𝑇𝑇𝑇 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑜𝑜𝑜𝑜 𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡 𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎
𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸𝐸 𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛𝑛 𝑜𝑜𝑜𝑜 𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡𝑡 𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑎𝑥𝑥100 
 
 


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9 
PHARMACOKINETICS 
The secondary endpoint PK parameters will be estimated using non compartmental methods from 
plasma concentration-time data. Refer to the PK Plan, Appendix 1, as prepared by SBQ.  
A formal analysis report will be prepared at the completion of the work and included in the CSR 
appendices. 
10 PHARMACODYNAMICS 
The secondary endpoint pSTAT3 inhibition ex-vivo on whole blood cells will be assessed using a 
validated ELISA. Refer to thePD analysis plan, Appendix 2, as prepared by TetraQ.   
A formal analysis report will be prepared at the completion of the work and included in the CSR 
appendices. 
 
 


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11 SAFETY  
11.1 Safety Endpoints 
Safety and tolerability will be assessed by clinical review of the following parameters:  
• 
AEs (including SAEs and AESIs) 
• 
Vital signs and other body measurements including respiratory rate and body temperature 
• 
12-lead ECG 
• 
Haematology, chemistry, coagulation and urinalysis 
• 
Physical examination 
All descriptive statistics for safety parameters will be evaluated using the Safety Set.    
11.2 Adverse Events 
11.2.1 Definitions 
• 
Treatment Emergent Adverse Event (TEAE) 
o A TEAE is defined as an AE that commences on, or after, the first administration of 
IMP.  AEs 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 
started and/or stopped prior to the first administration of IMP. 
• 
Adverse Events of Special Interest (AESI) 
o An adverse event of special interest (AESI) (serious or non-serious) is one of 
scientific and medical concern specific to the Sponsor’s product or programme, for 
which ongoing monitoring and rapid communication by the PI to the Sponsor is 
appropriate. 
11.2.2 Parameters 
• 
Event Term 
• 
Dates and times of onset and resolution 
• 
Severity (CTCAE Grades 1 to 5) 
• 
Outcome (Recovered/resolved, Recovering/resolving, Not recovered/not resolved, 
Recovered with sequelae/resolved with sequelae, Fatal, Unknown) 
• 
Relationship of AE to Ruxolitinib (Not Related, Related) 
• 
Relationship of AE to Artemether-lumefantrine (Not Related, Related) 
• 
Relationship of AE to Placebo (Not Related, Related) 
• 
Relationship of AE to Non-IMP Treatments / Study Procedures (Not Related, Related) 
• 
Action taken with Ruxolitinib, Artemether-lumefantrine and Placebo  (Dose not changed, 
Drug interrupted, Drug withdrawn, Not applicable, Unknown) 
• 
Other actions taken (including withdrawal from the study) 
• 
Seriousness (and SAE category) 
• 
AESI status (Yes/No) 
• 
Relatedness to COVID-19  
Derived parameters: 
• 
Duration in hours  
• 
Time of onset relative to first AL administration 
 
 


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11.2.3 Biostatistical methods 
Adverse events will be coded using MedDRA, and the MedDRA version will be included in the 
footnote of all AE listings and tables.  
11.2.3.1 Listings 
All AE data will be listed for each participant, including severity, relationship to IMPs, relationship to 
non-IMP protocol-specific treatments, outcome and actions taken. In addition, listings of AEs leading 
to discontinuation of the study, SAEs and deaths, will be provided as applicable. 
11.2.3.2 Tables 
AE summary tabulations will be restricted to TEAEs only.  
All reported TEAEs, including SAEs and AESIs, will be mapped to standard MedDRA coding terms 
and grouped by SOC and PT.   
An overview summary table of AEs will be provided by treatment arm including: 
− 
Number of events reported 
o Total TEAEs 
o Severe TEAEs 
o Artemether-lumefantrine Related TEAEs 
o Ruxolitinib Related TEAEs 
o Placebo Related TEAEs 
o Serious TEAEs 
- 
Number of participants reporting at least one of the following: 
o TEAE 
o Severe TEAE 
o Artemether-lumefantrine Related TEAEs 
o Ruxolitinib Related TEAEs 
o Placebo Related TEAEs 
o Serious TEAE 
− 
Number of participants withdrawn from the study due to a TEAE 
− 
Number of deaths 
The number of events, as well as the number and percentage of participants experiencing a TEAE, 
will be summarised for each SOC and PT by treatment arm and overall for the following categories 
of events: 
• 
All TEAEs 
• 
TEAEs by severity 
• 
TEAEs by relationship (separate tables for each relationship type assessed) 
• 
Serious TEAEs  
• 
TEAEs leading to study withdrawal  
• 
TEAEs of Special Interest (AESIs) 
For the summaries of TEAEs, participants who experience the same AE (in terms of the MedDRA 
preferred term) more than once will only be counted once. 
 
 


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11.3 Concomitant Medications 
Concomitant medications will be coded using the WHODRL, the version of which will be included in 
the footnote of all relevant listings and tables  
Concomitant medications are defined as medications continued or newly received at or after 
administration of AL, through to the End of Study visit. If a medication has a missing or partial 
missing start/end date or time and it cannot be determined whether it was taken before initial 
treatment or concomitantly, it will be considered as prior and concomitant. 
Concomitant medications will be summarised by Anatomical Therapeutic Chemical (ATC) (class 
level 3) and Preferred Name (PN). The summary tables will show the number and percentage of 
participants taking each medication by ATC and PN. 
Participants who take the same medication (in terms of the ATC and PN) more than once will only 
be counted once for that medication.  
11.4 Laboratory 
11.4.1 Parameters 
11.4.1.1 Hematology 
• Basophils (absolute) 
• Neutrophils (absolute) 
• Eosinophils (absolute) 
• Platelet count 
• Hematocrit 
• Red cell count (RCC) 
• Hemoglobin 
• Red cell distribution width (RDW) 
• Lymphocytes (absolute) 
• Reticulocyte count 
• Mean Cell Volume (MCV) 
• White cell count (WCC) 
• Monocytes (absolute) 
 
 
 
• Blood group and Rh(D) (Screening only) 
 Chemistry 
• Alanine Aminotransferase (ALT) 
• Estimated Glomerular Filtration Rate 
(eGFR, CKD-EPI equation) 
• Albumin 
• Gamma Glutamyl Transpeptidase (GGT) 
• Alkaline phosphatase (ALP) 
• Glucose (Fasted) 
• Aspartate Aminotransferase (AST) 
• Lactate dehydrogenase (LDH) 
• Bicarbonate 
• Magnesium 
• Bilirubin (direct) 
• Phosphate 
• Bilirubin (total) 
• Potassium 
• Calcium 
• Protein Total 
• Chloride 
• Sodium 
• Correct calcium 
• Urea 
• Creatinine 
• Uric acid 
• Creatine Kinase (CK) 
 
 
 
Lipids at Screening only 
 
• Cholesterol Total 
• Low Density Lipoprotein (LDL) 
• High Density Lipoprotein (HDL) 
• Triglyceride 
 
 
Pregnancy Test and FSH 
 
• β-Human Chorionic Gonadotropin (β-HCG) (women of child bearing potential only) 
• Follicular Stimulating Hormone (FSH) (post-menopausal women only) 


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 Urinalysis  
Dipstick Testing 
Quantiative Assessments 
Additional Assessments 
where urine dipstick result 
more than traces 
• Bilirubin 
• Creatinine 
• Bacteria 
• Blood (erythrocytes) 
• Glucose 
• Red blood cells 
• Glucose 
• Protein 
• White blood cells 
• Ketone bodies 
• Protein:Creatinine Ratio 
 
• Leucocytes 
 
 
• Nitrites 
 
 
• Proteins 
 
 
• pH 
 
 
• Specific gravity 
 
 
11.4.1.2 Coagulation 
• INR 
• Prothrombin Time 
• Activated partial thromboplastin time (APTT) 
11.4.1.3 Serology 
• Hepatitis B Surface Antigen (HBsAg) 
• Anti-HBc: IgG plus IgM if IgG is positive 
• Hepatitis C Antibody (anti-HCV) 
• Human Immunodefiniciency Virus (HIV) 1/2 (anti-HIV1 and anti-HIV2 Ab) 
11.4.1.4 Tuberculosis Testing 
• Latent M. tuberculosis infection testing using Quantiferon-TB gold assay 
11.4.2 Biostatistical methods 
All laboratory parameters will be listed with flags for values outside the reference ranges and values 
considered to be clinically significant by the investigator. 
Haematology, biochemistry (excluding β-HCG and FSH) and continuous urinalysis laboratory data 
will be summarised for each scheduled visit, including observed values and change from each 
baseline. Categorical urinalysis results will be summarised for each scheduled visit using frequency 
tabulations. 
Any available microscopic urinalysis will be listed.  
Cross-tabulations (shift tables) may be presented. 
11.5 Pregnancy Test and FSH 
Pregnancy test data will be listed for all women of child bearing potential. 
FSH data will be listed for all post-menopausal women.    
 
 


Protocol Number: MMV_Ruxolitinib_19_01  
16 December 2020 
Document status: SAP Version 1.0 (FINAL) 
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CONFIDENTIAL 
Page 20 of 22 
 
11.6 Vital Signs 
11.6.1 Parameters 
• 
Systolic Blood Pressure (SBP) (mmHg) 
• 
Diastolic Blood Pressure (DBP) (mmHg) 
• 
Heart Rate (beats/minute) 
• 
Tympanic Body Temperature (°C) 
• 
Respiratory Rate (breaths/minute) 
11.6.2 Biostatistical methods 
All vital signs data will be listed for all participants.  Any values outside of the protocol defined 
normal ranges (Table 1) will be flagged, with clinical significance status presented for out of range 
results. 
Table 1: Vital Signs Normal Ranges 
Parameter 
Range 
Systolic blood pressure             
90-140 mmHg 
Diastolic blood pressure           
50-90 mmHg 
Heart rate                                 
50-100 bpm 
Tympanic body temperature   
35.0-37.5°C 
Respiratory rate                        
10-25 breaths/min 
 
Vital sign parameters will be summarised by presenting summary statistics for observed values and  
change from baseline values for each scheduled visit and timepoint, with two separate tables for: 
• 
Vital Signs (SBP, DBP, heart rate) 
• 
Respiratory Rate and Temperature 
11.7 Physical Examination 
11.7.1 Parameters 
• Abdomen 
• Lungs (Respiratory) 
• Chest 
• Neck (including Thyroid) 
• General appearance 
• Neurological 
• Heart/Circulation 
• Skin 
• HEENT (Head, Eyes, Ears, Nose, Throat) 
• Other (free text) 
11.7.2 Biostatistical methods 
Physical examination parameters will be listed for all participants and visits. 
 
 


Protocol Number: MMV_Ruxolitinib_19_01  
16 December 2020 
Document status: SAP Version 1.0 (FINAL) 
BM S01-A, v2.0 
Statistical Analysis Plan 
Template Effective: 18/JAN/2018 
CONFIDENTIAL 
Page 21 of 22 
 
11.8 12-Lead ECG 
11.8.1 Parameters 
• 
The following ECG parameters are collected in triplicate for each timepoint: 
o PR interval (msec) 
o QRS interval (msec) 
o QT interval (msec) 
o QTcB interval (msec) 
o QTcF interval (msec) 
o Heart rate (beats/minute) 
o RR interval (msec) 
• 
ECG result (Normal, Abnormal – Not clinically significant, Abnormal – Clinically significant) 
• 
ECG abnormality (as appropriate) 
11.8.2 Biostatistical methods 
ECG parameters will be listed for all participants and time points. 
Observed values, as well as changes from each baseline, will be summarised descriptively for all 
ECG parameters by visit.   
An addition table will present the frequencies of participants who fulfill the following criteria, 
considering all scheduled and non-scheduled visit data: 
• 
QTcF prolongation >30 
• 
QTcB prolongation >30 
• 
QTcF prolongation >60 
• 
QTcB prolongation >60 
• 
QTcF >450 msec 
• 
QTcB >450 msec 
 
All ECG data will be listed for all participants.  Any values outside of the protocol defined normal 
ranges (Table 1) will be flagged. 
Table 2: ECG Normal Ranges 
Parameter 
Range 
PR interval 
≤210 msec 
QRS interval 
50–120 msec 
QTcB/QTcF       
≤450 msec 
11.9 Beck Depression Inventory 
Beck Depression Inventory (BDI-II) data will be listed for all participants, including answers to the 
individual questions as well as the total scores. 
 
 


Protocol Number: MMV_Ruxolitinib_19_01  
16 December 2020 
Document status: SAP Version 1.0 (FINAL) 
BM S01-A, v2.0 
Statistical Analysis Plan 
Template Effective: 18/JAN/2018 
CONFIDENTIAL 
Page 22 of 22 
 
12 HANDLING OF MISSING DATA 
Only recorded data will be analysed/presented and any participants who have missing data will only 
have observed data reported, with no imputation for missing data. 
13 CHANGES AND CLARIFICATIONS TO THE PLANNED ANALYSIS 
Not applicable. 
14 INTERIM ANALYSES 
This trial has no formal interim analyses other than data reviews of safety and tolerability as 
described in the clinical study protocol. 
15 SOFTWARE 
The following software will be used to perform the statistical analyses: Statistical Analysis System 
(SAS®) Version 9.4 (SAS Institute, Cary, North Carolina, USA). 
16 REFERENCES 
1) MMV_Ruxolitinib_19_01  Clinical Study Protocol, v4.0, 7 September 2020 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
1 of 13 
Confidential 
 
 
PHARMACOKINETIC ANALYSIS PLAN STUDY 
MMV_RUXOLITINIB_19_01 
 
 A randomised, single blind, placebo controlled, phase 1 trial to 
evaluate the safety, tolerability, pharmacokinetic and 
pharmacodynamic activity of ruxolitinib when co-administered 
with artemether-lumefantrine in healthy participants 
 
 
Investigational drugs: 
Ruxolitinib phosphate (Jakavi®) 
Artemether-lumefantrine (Riamet®) 
 
 
Clinical study number: 
MMV_RUXOLITINIB_19_01  
 
 
 
SBQ study number: 
SBQ-20251 
 
 
 
 
Study director: 
Paul van Giersbergen, PhD 
 
 
 
 
Test Facility: 
Swiss BioQuant AG 
Kägenstrasse 18, 4153 Reinach, Switzerland 
 
 
Sponsor: 
Medicines for Malaria Venture 
ICC, Route de Pre-Bois 20, 1215 Geneva 15, Switzerland 
 
 
 
Issue date: 
December 7, 2020 
 
 
Confidential 
The information contained in this document is the property of the sponsor of this study, 
Medicines for Malaria Venture (MMV). It may not be produced, published, or disclosed 
to others without written authorization of MMV. 


PK analysis plan
Protocol MMV_RUXOLITINIB 19 01
1. 
SIGNATURE PAGES
Study Director
Date: o? - Dec - 2.o2 o
-
Signature:W
* Paul van Giersbergen is a consultant for Swiss Bioeuant AG
Paul LM van Giersbergen, PhD*
Van Giersbergen Consulting
88 rue Principale
68500 Wuenheim
France
E-mail : paul.vangiersbergen@wanadoo.fr
Version status: final
Version date: December 7, 2020
2of13
Confidential




PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
4 of 13 
Confidential 
 
Sponsor 
 
 
Date:   
_______________________       
 
 
Signature:   _______________________ 
 
Farouk Chughlay, MD  
Medicines for Malaria Venture 
ICC, Route de Pre-Bois 20 
1215 Geneva 15 
Switzerland 
 
E-mail:chughlayf@mmv.org 
 
 
The Sponsor's responsible person confirms by his signature on the pharmacokinetic 
analysis plan that all subjects signed the informed consent prior to the start of analysis. 
Any withdrawal of the informed consent by the subject to analyze his/her samples should 
be communicated to Swiss BioQuant. 
 
 
 
09 Dec 2020
p.o.  Stephan Chalon, MD, PhD - MMV


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
5 of 13 
Confidential 
 
2. 
TABLE OF CONTENTS 
1. 
SIGNATURE PAGES .............................................................................................. 2 
2. 
TABLE OF CONTENTS ......................................................................................... 5 
3. 
ABBREVIATIONS AND DEFINITIONS .............................................................. 6 
4. 
INTRODUCTION ..................................................................................................... 7 
5. 
PERSONNEL / RESPONSIBILITY / COMMUNICATION ............................... 7 
5.1. 
SPONSOR .............................................................................................................. 7 
5.2. 
CLINICAL TEST FACILITY ..................................................................................... 7 
5.3. 
PHARMACOKINETIC TEST FACILITY (SWISS BIOQUANT AG) ............................... 7 
6. 
CLINICAL STUDY OBJECTIVES ........................................................................ 8 
7. 
CLINICAL STUDY DESIGN .................................................................................. 8 
7.1. 
PHARMACOKINETIC ASSESSMENTS ....................................................................... 9 
8. 
PLANNED PHARMACOKINETIC ANALYSIS .................................................. 9 
8.1. 
PHARMACOKINETIC ANALYSIS SET ....................................................................... 9 
8.2. 
DATA REVIEW ...................................................................................................... 9 
8.3. 
PROTOCOL DEVIATIONS ....................................................................................... 9 
8.4. 
TREATMENT COMPLIANCE ................................................................................. 10 
8.5. 
PREMATURE DISCONTINUATION AND MISSING DATA .......................................... 10 
8.6. 
HANDLING OF VALUES BELOW THE LIMIT OF QUANTIFICATION .......................... 10 
8.7. 
EXAMINATION OF SUBGROUPS AND COVARIATES ............................................... 10 
8.8. 
MEAN PHARMACOKINETIC PLOTS ...................................................................... 10 
8.9. 
INDIVIDUAL PHARMACOKINETIC PLOTS ............................................................. 10 
8.10. NONCOMPARTMENTAL PHARMACOKINETIC ANALYSIS ....................................... 11 
8.11. STATISTICS......................................................................................................... 12 
8.12. SOFTWARE ......................................................................................................... 12 
9. 
REPORTING CONVENTIONS ............................................................................ 12 
10. DATA QUALITY ASSURANCE .......................................................................... 12 
11. ARCHIVING ........................................................................................................... 12 
12. REFERENCES ........................................................................................................ 13 
 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
6 of 13 
Confidential 
 
3. 
ABBREVIATIONS AND DEFINITIONS 
AL 
Artemether-lumefantrine 
AUC0-∞ 
Area under plasma concentration-time curve from time zero, i.e., 
predose on Day 1, to infinity 
AUC0-t 
Area under plasma concentration-time curve from time zero, i.e., 
predose on Day 1, to time t of the last measured concentration above 
the limit of quantification after the last dose on Day 3 
AUC0-6h 
Area under plasma concentration-time curve from time zero to time 
6h after dosing with Rux on Day 1 
AUC0-8h 
Area under plasma concentration-time curve from time zero to time 
8h after dosing with AL on Day 1 
AUC0-10h 
Area under plasma concentration-time curve from time zero to time 
6h after dosing with Rux on Day 3 
AUC0-12h 
Area under plasma concentration-time curve from time zero to time 
8h after dosing with AL on Day 3 
BLQ 
Below the limit of quantification 
Cmax 
Maximum plasma concentration 
CV 
Coefficient of variation 
λz 
First order rate constant associated with the terminal log-linear 
portion of the plasma concentration-time curve 
MMV 
Medicines for Malaria Venture 
P. falciparum 
Plasmodium falciparum 
PK 
Pharmacokinetic(s) 
R2 
Linear regression coefficient 
Rux 
Ruxolitinib 
SD 
Standard deviation 
t1/2 
Apparent terminal half-life 
tmax 
Time to reach maximum plasma concentration 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
7 of 13 
Confidential 
 
4. 
INTRODUCTION 
The clinical study MMV_RUXOLITINIB_19_01 was conducted as part of the ruxolitinib 
(Rux)/artemether-lumefantrine (AL) clinical development program. The study population 
comprised of adult male and female participants aged 18 to 55 years.  
In malaria volunteer infection studies, type I interferons s produced by several 
different cell sources were found to be important regulators of developing anti-parasitic 
immunity. Type I interferons are key immunomodulatory molecules in humans infected 
with P. falciparum, and targeting this pathway for clinical advantage is a promising 
strategy to overcome established or developing immunoregulatory networks, to improve 
natural, drug-mediated, or vaccine-induced immunity against malaria. Rux blocks type 1 
interferon signaling, and is, therefore, a candidate for investigation in combination with 
established anti-malarial medications such as AL. 
Study MMV_RUXOLITINIB_19_01 is the first study in humans to investigate the 
tolerability, safety, pharmacokinetics (PK) and pharmacodynamics of the combination of 
Rux and AL.  
5. 
PERSONNEL / RESPONSIBILITY / COMMUNICATION 
First choice of communication is via e-mail. 
5.1. 
Sponsor 
Project Director: 
Farouk Chughlay, MD  
MMV, ICC, Route de Pre-Bois 20, 1215 Geneva 15, 
Switzerland  
Tel.: +41 (0) 79 123 4289 
Fax: +41 (0) 22 799 4061 
e-mail: chughlayf@mmv.org 
 
5.2. 
Clinical Test Facility 
Principal Investigator: 
Paul Griffin, MD  
Q-Pharm Pty Ltd 
Level 5, 300C Herston Rd, Herston Qld 4006 
Australia 
 
5.3. 
Pharmacokinetic Test Facility (Swiss BioQuant AG) 
Study Director: 
Paul LM van Giersbergen, PhD 
Tel.: +33 3 89 76 77 75 
e-mail: paul.vangiersbergen@wanadoo.fr 
 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
8 of 13 
Confidential 
 
Quality Assurance Contact: Larissa Michel 
QA department at Swiss BioQuant AG 
Tel.: +41 (0) 61 71698 19 
Fax: +41 (0) 61 71698 15 
e-mail: qau@swissbioquant.com 
Head of Test Facility: 
Christoph Siethoff, PhD 
Tel.: +41 (0) 61 71698 12 
Fax: +41 (0) 61 71698 15 
e-mail: christoph.siethoff@swissbioquant.com 
6. 
CLINICAL STUDY OBJECTIVES 
Primary objective: 
• To assess the safety and tolerability of 3-day b.i.d dosing of AL+Rux and 
AL+placebo. 
Secondary objectives: 
• To assess the effect of AL+Rux and AL+placebo on pSTAT3 inhibition.  
• To characterise the PK profiles of artemether and its major metabolite 
dihydroartemisinin [DHA], lumefantrine, and Rux.  
• To assess additional safety variables (tympanic body temperature and respiration rate)  
7. 
CLINICAL STUDY DESIGN 
This study is a randomised, single-blind, placebo-controlled, single centre phase 1 trial. 
Subjects will be administered one of two treatments:  
• AL ( 20 mg artemether /120 mg lumefantrine) and Rux (20 mg), or 
• AL ( 20 mg artemether /120 mg lumefantrine) and placebo  
The study will be composed of 2 groups (Group 1a and Group 1b) to be enrolled 
sequentially. Group 1a will participate first as the sentinel safety group. 
Eight participants will be randomized to receive AL+Rux or AL+placebo in a 3:1 ratio 
overall (1:1 in Group 1a, and 5:1 in Group 1b). Participants will be administered oral 
doses of AL, with oral doses of Rux or placebo administered 2 hours later twice daily 
(b.i.d) for 3 consecutive days (AL at t = 0, 8, 24, 36, 48 and 60 hours, Rux at t = 2, 10, 
26, 38, 50 and 62 hours).  
A more detailed description of study design and study procedures is provided in the 
clinical study protocol [1]. 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
9 of 13 
Confidential 
 
7.1. 
Pharmacokinetic assessments 
Blood samples for PK will be collected at the following times:  
• Day 1 prior to commencement of dosing (pre-dose) and at t = 1, 2, 3, 4, 5, 6, 8, 12, 24, 
36, 48, 60, 62, 64, 72 and 78 hours post commencement of dosing. For time points 
coinciding with study drug administration, blood sampling occurs prior to study drug 
administration. 
• Days 8, 11, 15, 21, 24 and 27 at t = 168, 240, 336, 480, 552 and 624 hours post 
commencement of dosing (±120 min for t = 168 hours, ± 24 hours for later 
timepoints). 
• Day 29 at t = 672 hours (± 24 hours). 
The exact date and time of blood sampling will be recorded in the Case Report Form. 
The plasma concentrations of Rux, AL and the artemether metabolite dihydroartemisinin 
will be measured using validated liquid chromatography-tandem mass spectrometry 
methods at Swiss BioQuant AG, Reinach, Switzerland under phase number SBQ-20227.     
8. 
PLANNED PHARMACOKINETIC ANALYSIS 
8.1. 
Pharmacokinetic analysis set 
All subjects who received a dose of study medication will be included in the 
pharmacokinetic analysis provided they have at least one available Rux and/or AL 
plasma concentration above the limit of quantification and who experienced no protocol 
deviations with relevant impact on the PK data. The number of subjects included into the 
PK analysis will be listed.   
8.2. 
Data review 
The exclusion of outliers, subjects and/or data points from analyses will not be done on a 
purely statistical basis. If a separate analysis was carried out excluding outliers, it would 
not normally be included in the PK report unless the conclusions differ from the primary 
analysis. 
Concentration values excluded from the calculation of PK variables will be flagged. The 
pharmacokineticist will provide a reason in the PK report for excluding any points or 
subjects. The dataset for PK analysis will be created by Van Giersbergen Consulting 
(consultant for Swiss BioQuant AG) using electronic data files provided by Swiss 
BioQuant AG (bioanalytical data with scheduled blood sampling times) and MMV 
(listings of demography, protocol violations, adverse events and actual blood sampling 
times in hour). The dataset will be added in the report as a listing. 
8.3. 
PK analysis plan deviations 
Deviations to this PK analysis plan analysis will be listed.   


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
10 of 13 
Confidential 
 
8.4. 
Treatment compliance 
Treatment compliance is assumed 100% since study drug is administered by study 
personel and will not be used in the analysis. A listing of dosing will, however, be 
provided. 
8.5. 
 Premature discontinuation and missing data 
Subjects who did not receive any study drug or who did not receive the full treatment will 
be excluded from the PK analysis.  Subjects who prematurely discontinued the study 
resulting in incomplete PK profiles will be included in the PK analysis but will be 
excluded from descriptive statistics. Missing sample concentrations will be ignored, i.e. 
no imputation of missing concentration data will be performed. 
8.6. 
Handling of values below the limit of quantification  
Samples with plasma concentrations below the limit of quantification (BLQ) will be set 
to zero for the PK analysis. In the dataset listing, they will be reported as BLQ. BLQ 
values occurring between two measurable concentrations will be set to missing. 
8.7. 
Examination of subgroups and covariates 
No subgroup/covariate PK analysis is foreseen in the protocol.  
8.8. 
Mean pharmacokinetic plots 
Nominal blood sampling times will be used to calculate the mean drug concentrations at 
each time point.   
Comparative linear and semi-logarithmic plots of the mean concentration-time data for 
each analyte and corresponding tables of descriptive statistics (mean, SD, minimum, 
median, maximum) will be prepared. 
In addition, plots of trough mean concentration-time data for AL and Rux will be 
prepared and used for a graphical exploration. 
For plots and summary tables, BLQ values will be set to zero. 
8.9. 
Pharmacokinetic plots 
Linear and semi-logarithmic plots for each subject will be prepared using actual sample 
times. The time points used for the estimation of t1/2, if applicable, will be shown on the 
semi-logarithmic plots as well as the coefficient of the linear regression, R2, used to 
calculate t1/2. In addition, plots on a linear and semi-logarithmic scale will be prepared 
showing all individual concentration-time profiles per treatment in one plot. 
For individual plots, BLQ values will be handled as described in section 8.6. 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
11 of 13 
Confidential 
 
8.10. 
Noncompartmental pharmacokinetic analysis  
The PK variables of Rux, AL and the artemether metabolite dihydroartemisinin will be 
computed using the actual sampling times relative to the time of study drug 
administration.   
Cmax, tmax, C168h 
The maximum observed drug concentration (Cmax) and the first time of its occurrence 
(tmax) will be obtained directly from the concentration-time data after the first and last 
administration of study drugs. For lumefantrine only, C168h obtained directly from 
concentration-time data, will be separately listed. 
λz, t1/2 
• The terminal phase rate constant (λz) will be estimated by linear regression of 
logarithmically transformed concentration versus time data.  
• Only those data points which are judged to describe the terminal log-linear decline 
will be used in the regression.   
• A minimum number of three data points will be used in calculating λz.  
• λz will not be estimated if R2 
value of the linear regression is less than 0.8. As a 
consequence, λz -dependent pharmacokinetic variables will not be estimated. 
• Further, it is preferable that the duration of time over which λz is estimated be at least 
two times the subsequently estimated terminal phase half-life (t1/2).  
The t1/2 will be calculated as follows: t1/2=ln2/λz. 
The concentration-time data used for estimation of the terminal phase rate constant will 
be visible on the individual semi-logarithmic plasma concentration-time profiles. 
AUC0-t, AUC0-inf, AUC0-6h, AUC0-8h, AUC0-10h, AUC0-12h 
The area under the concentration-time curve (AUC) from time zero, i.e., predose AL on 
Day 1, to time t of last quantifiable concentration (AUC0-t) after the last study drug 
administration on Day 3. The area from time zero extrapolated to infinite (AUC0-inf) will 
be calculated as follows: AUC0-inf = AUC0-t + Ct/λz where Ct is the last measured 
concentration above the lower limit of quantification. 
On Day 1, after the first dose of study drug in the morning, AUC0-8h will be calculated for 
AL and AUC0-6h for Rux whereby time 0 refers to predose of the respective compound. 
On Day 3, after the second dose of study drug in the evening, AUC0-12h will be calculated 
for AL and AUC0-10h for Rux whereby time 0 refers to predose of the respective 
compound. 
The different AUCs will be calculated using the linear up log down trapezoidal method. 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
12 of 13 
Confidential 
 
8.11. 
Statistics 
Listings of individual PK variables for Rux, AL and the artemether metabolite 
dihydroartemisinin will be provided. 
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 treatment.  
8.12. 
Software 
The data file, which will be the basis for the PK analysis, will be created using Excel. It 
will contain the subjects' identifier, dose administered, scheduled and actual timepoints of 
PK blood sampling, concentration data, etc. 
The PK analysis and the statistics will be performed with Phoenix WinNonlin (version 
8.2, Pharsight Corporation, Mountain View, CA, USA).  
The individual plasma concentration time profiles will be generated with Phoenix 
WinNonlin whereas other plots will be generated with GraphPad Prism (version 5.00, 
GraphPad Software Inc, La Jolla, CA, USA). 
9. 
REPORTING CONVENTIONS 
The PK analysis will be reported using a Van Giersbergen Consulting report template 
(Word). As a rule, 3 significant digits will be used in the reporting of the results of the PK 
analysis. 
The final report will comprise a QA statement of the Quality Assurance of Swiss 
BioQuant AG. 
The final report will be provided in English. An electronic version (searchable, with 
hyperlinked table of contents and cross-references) of the PK analysis report will be 
provided in current PDF format to the Sponsor.  
10. 
 DATA QUALITY ASSURANCE 
The Quality Assurance department of Swiss BioQuant AG (Reinach, Switzerland) will 
inspect the PK analysis report. 
11. 
ARCHIVING 
All documents (WinNonlin reports, QA protocols, etc.) generated by Van Giersbergen 
Consulting and the final PK analysis report will be filed in the Swiss BioQuant AG 
archives for 10 years. 


PK analysis plan 
 
Protocol MMV_RUXOLITINIB_19_01 
Version status: final 
Version date:  December 7,  2020 
13 of 13 
Confidential 
 
At the end of the mentioned storage period Swiss BioQuant AG will ask the Sponsor for 
the authorization to destroy the documents of the study or to transfer them to the 
Sponsor’s facility. 
12. 
REFERENCES 
[1] 
A randomised, single blind, placebo controlled, phase 1 trial to evaluate the safety, 
tolerability, pharmacokinetic and pharmacodynamic activity of ruxolitinib when co-
administered with artemether-lumefantrine in healthy participants. Protocol number 
MMV_RUXOLITINIB_19_01, version 4, dated 07 September 2020. 


P a g e  |  1  
 
 
 
 
Pharmacodynamic and Statistical Analysis Plan 
 
 
Product: 
Ruxolitinib phosphate 
 
Artemether-lumefantrine 
 
Project Title: 
Protocol MMV_Ruxolitinib_19_01 
A randomised, single-blind, placebo controlled, phase 1 trial to 
evaluate 
the 
safety, 
tolerability, 
pharmacokinetic 
and 
pharmacodynamic activity of ruxolitinib when co-administered with 
artemether-lumefantrine in healthy participants 
 
Sponsor: 
Medicines for Malaria Venture (MMV) 
 
20 Route de Pré-Bois 
 
1215 Geneva 15 
 
SWITZERLAND 
 
PK/PD Consultant: 
Stephanie Reuter Lange, PhD BSc (Hons) 
 
UniSA Clinical & Health Sciences 
 
University of South Australia 
 
CEA-19, GPO Box 2471 
 
Adelaide, SA, 5001 
 
Phone: +61 8 8302 1872 
 
Email: stephanie.reuterlange@unisa.edu.au 
 
Bioanalytical Test Facility: 
TetraQ 
 
Level 7, Block 6 
 
Royal Brisbane and Women’s Hospital 
 
Herston QLD 4029 
 
Version: 
FINAL 
 
Release Date: 
16 December 2020 
 


Pharmacodynamic and Statistical Analysis Plan 
CONFIDENTIAL 
Protocol MMV_Ruxolitinib_19_01 
FINAL: 16 December 2020 
P a g e  |  2  
This Pharmacodynamic and Statistical Analysis Plan has been prepared for the study entitled “Protocol 
MMV_Ruxolitinib_19_01: A randomised, single blind, placebo controlled, phase 1 trial to evaluate the safety, 
tolerability, pharmacokinetic and pharmacodynamic activity of ruxolitinib when co-administered with 
artemether-lumefantrine in healthy participants”. The contents of this document are confidential and 
reproduction, in whole or part, is not permitted without prior written permission from TetraQ. 
 
 
 
 __________________________________________________ 
Dr Stephanie E Reuter Lange 
Pharmacokinetic/Pharmacodynamic Consultant
 
 
 
This Pharmacodynamic and Statistical Analysis Plan has been reviewed and approved by representatives on 
behalf of the Medicines for Malaria Venture (MMV). 
 
 
 
 __________________________________________________ 
Dr Farouk Chughlay 
Medical Director, Experimental Medicine & Clinical Pharmacology 
 
 
 
 __________________________________________________ 
Dr Myriam El Gaaloul 
Director, Clinical Sciences 
 
 
 
Stephanie Reuter Lange 
cn=Stephanie Reuter Lange, 
o=University of South 
Australia, ou=UniSA Clinical 
and Health Sciences, 
email=stephanie.reuterlange@
unisa.edu.au, c=AU 
2020.12.16 20:10:48 +10'30'
Stephan
Chalon
Digitally signed by 
Stephan Chalon 
Date: 2020.12.16 
11:03:44 +01'00'
Myriam El 
Gaaloul
Digitally signed by Myriam El 
Gaaloul
DN: cn=Myriam El Gaaloul, 
o=MMV, ou, 
email=elgaaloulm@mmv.org,
c=CH
Date: 2020.12.16 13:51:15 
+01'00'


Pharmacodynamic and Statistical Analysis Plan 
CONFIDENTIAL 
Protocol MMV_Ruxolitinib_19_01 
FINAL: 16 December 2020 
P a g e  |  3  
1. Introduction 
Study Rationale 
Medicines for Malaria Venture (MMV) is conducting a clinical trial program with ruxolitinib as a therapeutic 
option in the treatment of malaria. This phase I, multiple-dose study is being conducted to evaluate the 
safety, tolerability, pharmacokinetics, and pharmacodynamics of artemether-lumefantrine + ruxolitinib and 
artemether-lumefantrine + placebo in healthy volunteers. 
Study Objectives 
(i) 
To assess the safety and tolerability of 3-day b.i.d. dosing of artemether-lumefantrine + ruxolitinib 
and artemether-lumefantrine + placebo 
(ii) 
To assess the effect of artemether-lumefantrine + ruxolitinib and artemether-lumefantrine + 
placebo on pSTAT3 inhibition 
(iii) 
To characterise the pharmacokinetic profile of artemether and its major metabolite 
dihydroartemisinin, lumefantrine and ruxolitinib 
(iv) 
To assess additional safety variables (tympanic body temperature and respiration rate) 
The proposed pharmacodynamic analysis will be conducted to assist in addressing objective (ii). 
Study Design 
This is a randomised, single-blind, placebo-controlled, multiple-dose study comparing artemether-
lumefantrine + ruxolitinib and artemether-lumefantrine + placebo administered twice-daily over a 3-day 
period to a total of 8 participants (Group 1a [Sentinel] = 2 participants, Group 1b = 6 participants). 
Artemether-lumefantrine (20 mg artemether/120 mg lumefantrine) will be administered with 250 mL full-
fat milk at 0, 8, 24, 36, 48 and 60 hours, and ruxolitinib (20 mg) or placebo will be administered with 250 mL 
water at 2, 10, 26, 38, 50 and 62 hours. 
Blood samples for quantification of plasma ruxolitinib, artemether, dihydroartemisinin and lumefantrine 
concentrations will be collected prior to dosing (0 hr) and 1, 2, 3, 4, 5, 6, 8, 12, 24, 36, 48, 60, 62, 64, 72, 78, 
168, 240, 336, 480, 552, 624 and 672 hours after administration of artemether-lumefantrine on Day 1. 
Additionally, blood samples for determination of pSTAT3 levels will be collected prior to dosing (0 hr) and 
1, 2, 3, 4, 5, 6, 8, 12 and 168 hours after administration of artemether-lumefantrine on Day 1. Samples 
scheduled for the same time as dose administration will be collected immediately prior to dosing. 
2. Pharmacodynamic Analysis 
pSTAT3 Data 
All individual participant pSTAT3 levels and ruxolitinib concentrations will be presented within the 
Pharmacodynamic Consultancy Report in both tabular and graphical form. In addition, %pSTAT3 inhibition 
will be determined at each time-point, calculated as: 
%𝑝𝑆𝑇𝐴𝑇3 𝐼𝑛ℎ𝑖𝑏𝑖𝑡𝑖𝑜𝑛 = 100 × 𝑝𝑆𝑇𝐴𝑇30  − 𝑝𝑆𝑇𝐴𝑇3𝑖
𝑝𝑆𝑇𝐴𝑇30
 


Pharmacodynamic and Statistical Analysis Plan 
CONFIDENTIAL 
Protocol MMV_Ruxolitinib_19_01 
FINAL: 16 December 2020 
P a g e  |  4  
where, pSTAT30 is the pSTAT3 level prior to treatment initiation (i.e. at 0 hr), and pSTAT3i is the pSTAT3 level 
at the ith time-point. %pSTAT3 inhibition data will be presented within the Pharmacodynamic Consultancy 
Report in both tabular and graphical form 
Descriptive statistics (including mean, median, standard deviation, minimum, maximum, coefficient of 
variation and geometric mean) will be calculated for pSTAT3 levels and %pSTAT3 inhibition at each nominal 
time-point. Summary data will be tabulated and presented graphically. 
Pharmacodynamic Data 
Pharmacodynamic analysis will be based on %pSTAT3 inhibition data. For each individual, %pSTAT3 
inhibition data will be used for the calculation of: 
AUECT 
Area under the pharmacodynamic effect versus time profile over the ruxolitinib dosing interval 
on Day 1 (i.e. from 2 to 10 hours), calculated using the linear trapezoidal method. %pSTAT3 
inhibition levels at 10 hours will be interpolated based on values obtained at 8 and 12 hours. 
The pharmacokinetic/pharmacodynamic relationship between ruxolitinib concentrations and %pSTAT3 
inhibition will also be calculated using sigmoidal curve fitting according to the following equation: 
𝐼 = 𝐼𝑚𝑎𝑥 × 𝐶𝛾
𝐶𝛾 + 𝐼𝐶50
𝛾 
where, I is the %pSTAT3 inhibition, C is the ruxolitinib concentration, Imax is the theoretical maximum 
%pSTAT3 inhibition, IC50 is the ruxolitinib concentration at which there is 50% maximal inhibition, and γ is 
the Hill coefficient. All %pSTAT3 inhibition values and ruxolitinib concentrations at coinciding time-points 
will be included in the analysis. Alternate pharmacodynamic models may be investigated at the discretion 
of the Pharmacokinetic/Pharmacodynamic Consultant. 
All individual pharmacodynamic data will be presented within the Pharmacodynamic Consultancy Report in 
tabular form.  
Descriptive statistics (including mean, median, standard deviation, minimum, maximum, coefficient of 
variation and geometric mean) will be calculated for pharmacodynamic data and will be tabulated. 
Statistical Analysis 
A linear mixed-effects analysis of variance (ANOVA) will be used to perform statistical comparisons of 
Ln-transformed AUECT, IC50 and Imax data between study treatments. The residual error (error mean square) 
will be used to construct the 90% confidence intervals for the ratio of treatment means. 
No difference will be concluded if the 90% confidence intervals are within the standard regulatory limits of 
80-125%. Significance will be set at an α-level of 0.05. 
Statistical data will be presented within the Pharmacodynamic Consultancy Report in tabular form. 
3. Data Handling 
For graphical presentation of data, calculation of descriptive statistics and pharmacodynamic analysis, 
negative %pSTAT3 inhibition values will be designated a value of 0%. Plasma ruxolitinib concentrations 
below the limit of quantification of the assay will be designated a value of 0.00ng/mL for the 
pharmacokinetic/pharmacodynamic analysis. 


Pharmacodynamic and Statistical Analysis Plan 
CONFIDENTIAL 
Protocol MMV_Ruxolitinib_19_01 
FINAL: 16 December 2020 
P a g e  |  5  
When available, all times used in the presentation of individual pSTAT3 data and pharmacodynamic analysis 
will be actual times relative to first ruxolitinib administration on Day 1, with the exception of the pre-dose 
samples which will be designated a nominal time of 0.00 hr. In the event that actual sample collection times 
are not available, nominal times shall be used. 
All data will be reported to 3 significant figures.  
Non-rounded data will be used in descriptive, pharmacodynamic and statistical analyses.  
4. Analysis Packages 
Pharmacodynamic and statistical analyses will be conducted using Phoenix® WinNonlin® Version 8.0 or 
higher (Pharsight®, a Certara™ company). Internal validation of Phoenix® WinNonlin® will be conducted 
using the Phoenix® WinNonlin® Validation Suite prior to data analysis. 
5. Data Transfer 
Individual samples will be analysed for pSTAT3 levels by the TetraQ Analytical Laboratory. Ruxolitinib 
concentrations will be analysed by an external laboratory. Data will be provided to the 
Pharmacokinetic/Pharmacodynamic Consultant in Microsoft Excel format. 
Clinical data will be exported from the locked study database and transferred to the Pharmacodynamic 
Consultant in Microsoft Excel format and shall include at least the following information for each participant: 
identification code, treatment administration details and sample collection details. 
All pharmacodynamic data listings, tables and profiles will be included in the Pharmacodynamic 
Consultancy Report. Copies shall also be provided to the sponsor in Microsoft Word format for preparation 
of the Clinical Study Report.