The Effect of Dolutegravir on Whole-body Insulin Sensitivity, Lipid and Endocrine Profile in Healthy Volunteers
Phase: PHASE1
Conditions: HIV-1-infection on Healthy Volunteers
Sponsor: Chelsea and Westminster NHS Foundation Trust

 
 
 
 
 
 
 
CLINICAL TRIAL PROTOCOL 
 
 
 
 
 
Protocol Title: 
The effect of dolutegravir on whole-body insulin sensitivity, 
lipid and endocrine profile in healthy volunteers  
 
 
 
Protocol Number: 
 
CRF003 
 
Version 
 
 
Version 4.0 20 Jan 2023 
 
EudraCT No: 
 
 
2020-004395-17  
 
IRAS Number:    
 
274870 
 
Clinical Trial.gov:   
 
NCT04771754 
 
 
 
 
Chief Investigator:  
 
Dr Ruth Byrne  
St. Stephen’s Centre 
Chelsea and Westminster Hospital NHS Foundation Trust 
369 Fulham Road 
SW10 9NH London 
 
Sponsor:  
 
 
Chelsea and Westminster Hospital NHS Foundation Trust 
369 Fulham Road 
SW10 9NH London 
 
 
 
 
 
 
 
 
 
 
 
Protocol Approval:  
 
                   20 Jan 2023 
 
 
 
Dr Ruth Byrne  
      dd mmm yyyy 
 
 
 
 
 
GCP Compliance Statement: 
This trial will be conducted in compliance with the protocol, the principles that have their 
origin in the Declaration of Helsinki and all applicable regulatory requirements 

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CRF003 protocol version 4.0 20 Jan 2023 
Synopsis 
 
Name of Investigational Product: 
Dolutegravir 
Name of Active Ingredients: 
Dolutegravir 
Study Title: 
The effect of dolutegravir on whole-body insulin 
sensitivity, lipid and endocrine profile in healthy 
volunteers  
Phase of Study: 
Phase I 
Objectives: 
The objectives of this study are: 
Primary 
To quantify the difference in change in insulin sensitivity 
(determined by peripheral glucose uptake using a 
euglycaemic clamp) with the administration of 
dolutegravir (DTG) compared to no DTG for 28 days in 
HIV seronegative healthy volunteers.  
Secondary  
To investigate the effect of the dolutegravir on: 
1. Adipocytokines by assessing adiponectin and 
leptin levels 
2. Ghrelin  
3. Pituitary hormones  
4. Lipid profile including lipid fractions  
5. Changes in indirect calorimetry  
6. Changes in food intake by food diaries  
7. Change in sleep parameters by sleep 
questionnaires  
8. Safety and tolerability of dolutegravir 

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CRF003 protocol version 4.0 20 Jan 2023 
Study Design 
Randomised, crossover 
Indication: 
HIV negative volunteers 
Methodology: 
Insulin sensitivity determined by glucose disposal during 
steady state of euglycaemic clamp. 
Planned Sample Size: 
16 
Summary of Eligibility Criteria: 
 
HIV negative subjects 
 
Clinically well 
 
BMI between 18 and  30kg/m2  
 
Free from diabetes mellitus and  metabolic 
syndrome  
 
Not receiving any metabolically active medication or 
drugs likely to interact with antiretroviral therapy 
 
Able and willing to provide informed consent 
 
Number of Study Centres 
One 
Duration of Treatment: 
28 days per sequence with washout between sequences 
Dose and Route of Administration: 
Orally administered dolutegravir 50 mg once daily  
Criteria for Evaluation: 
Subjects will undergo three euglycaemic clamp 
procedures in order to determine the extent of glucose 
disposal. The first clamp will be performed prior to the 
commencement of the first study drug 
administration/no treatment, the second one following 
four weeks of study drug/no treatment, the third after 
the crossover phase to study drug/no treatment.    
Primary Endpoint: 
Change from baseline in total body glucose disposal by 
euglycaemic clamp method following 28 days treatment 

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Secondary Endpoints: 
 
Change from baseline in serum levels of 
fasting cholesterol fractions and 
triglycerides 
 
Changes from baseline in serum 
adipocytokines, ghrelin and pituitary 
hormones  
 
Changes from baseline in indirect 
calorimetry  
 
Caloric intake by food diaries 
 
Quality of sleep by  sleep questionnaires  
 
Safety data including the number and 
severity of adverse events, as assessed by 
the medical dictionary for regulatory 
activities (MedDRA), from baseline to day 
100  
 
 
 
 

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General Study Information 
 
Name of sponsor: 
Chelsea and Westminster Hospital NHS Foundation Trust 
369 Fulham Road 
LondonSW10 9NH 
Chief investigator: 
 
 
 
 
 
Co-Investigator 
Dr Ruth Byrne Chelsea and Westminster Hospital 
369 Fulham Road,  
London SW10 9NH 
Phone 0208 846 6507 
 
Dr Graeme Moyle  
Chelsea and Westminster Hospital 
369 Fulham Road,  
London SW10 9NH 
Phone 0208 846 6507 
Laboratory facilities:  
Biochemistry & Haematology  
 
 
Imperial College NHS Foundation Trust 
Pathology Laboratory 
Chelsea and Westminster Hospital 
369 Fulham Road 
London SW10 9NH 
Virology & Hepatitis 
Imperial College NHS Foundation Trust  
Dept. of Infection & Immunity 
9th Floor Laboratory Block 
Charing Cross Hospital 
Fulham Palace Road 
London W6 8RF 

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CRF003 protocol version 4.0 20 Jan 2023 
Medical expertise: 
Dr Ruth Byrne & Dr Graeme Moyle  
Chelsea and Westminster Hospital 
369 Fulham Road,  
London SW10 9NH 
 
Phone 0208 846 6507 
Data management and Statistics: Research and Development Office 
Chelsea and Westminster Hospital NHS Foundation Trust 
Unit G2, Harbour Yard 
Chelsea Harbour 
London SW10 0XD  
Monitor:  
Research and Development Office 
Chelsea and Westminster Hospital NHS Foundation Trust 
Unit G2, Harbour Yard 
Chelsea Harbour 
London SW10 0XD  
Phone 020 3315 6825 
Email: research.development@chelwest.nhs.uk  
 
 
 
 

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CRF003 protocol version 4.0 20 Jan 2023 
List of abbreviations and definitions of terms 
 
ACTG 
AIDS Clinical Trial Group 
AE 
Adverse event 
AIDS 
Acquired Immune Deficiency Syndrome 
ALT 
Alanine aminotransferase 
AST 
Aspartate aminotransferase 
BMI 
Body Mass Index 
CM 
Concomitant Medication 
CRF 
Case Report Form 
DAIDS 
Division of AIDS 
ECG 
Electrocardiogram 
FBC 
Full blood count 
GCP 
Good Clinical Practice 
GP 
General Practitioner 
HBV 
Hepatitis B virus 
HCV 
Hepatitis C virus 
HIV 
Human Immunodeficiency Virus 
ICH-GCP 
International conference of harmonization good clinical practice 
IMP 
Investigational Medicinal Product 
LFT 
Liver function test 
MHRA 
Medicines and Healthcare products Regulatory Agency 
NRTI 
Nucleoside reverse transcriptase inhibitor 
NNRTI 
Non-nucleoside reverse transcriptase inhibitor 
OD 
Once daily 
PI 
Protease inhibitors 

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CRF003 protocol version 4.0 20 Jan 2023 
PLWH 
People living with HIV 
SAE 
Serious Adverse Event 
SmPC 
Summary of product characteristics 
SUSAR 
Suspected Unexpected Serious Adverse Reaction 
TOPS 
The over-volunteering prevention scheme 
UK 
United Kingdom 
WOCBP 
Women of Child Bearing Potential 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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TABLE OF CONTENTS 
 
1. 
BACKGROUND INFORMATION ......................................................................................... 11 
1.1 Introduction & background .................................................................................................. 11 
1.2 Rationale ................................................................................................................................. 11 
2. 
TRIAL OBJECTIVES ............................................................................................................ 12 
2.1 
Primary Objective ........................................................................................................... 12 
2.2   
Secondary Objectives .................................................................................................... 12 
3. 
TRIAL DESIGN ..................................................................................................................... 12 
3.1 
Endpoints ........................................................................................................................ 12 
3.2 
Study design ................................................................................................................... 13 
3.3 
Randomisation ................................................................................................................ 13 
3.4  
Benefit: Risk Assessment ............................................................................................. 14 
4. 
SUBJECT POPULATION ..................................................................................................... 14 
4.1  
Number of subjects and subject selection .................................................................. 14 
4.1.1 
Inclusion Criteria ............................................................................................................. 14 
4.1.2 
Exclusion Criteria ............................................................................................................ 16 
4.2  
Duration of Involvement ................................................................................................ 17 
4.3 
Withdrawal of Subjects and Discontinuation Criteria ................................................. 17 
5. 
TREATMENT OF SUBJECTS .............................................................................................. 18 
5.1. Study Treatment Regimen ................................................................................................... 18 
5.2 
Treatment Compliance ................................................................................................... 18 
6. 
VISIT SCHEDULE & PROCEDURES .................................................................................. 18 
6.1 
Euglycaemic Clamp ........................................................................................................ 23 
6.2 Indirect Calorimetry ............................................................................................................... 24 
7 
STATISTICAL ANALYSIS ................................................................................................... 24 
7.1 
Sample size ..................................................................................................................... 24 
7.2   Criteria for evaluation ......................................................................................................... 25 
7.3 
Statistical Methods ......................................................................................................... 25 
7.3.1 Efficacy parameters ............................................................................................................. 26 
7.3.1 Safety parameters ............................................................................................................... 26 
7.4 
Criteria for termination of the trial ................................................................................ 26 
7.5 Toxicity Management ............................................................................................................ 26 
7.6 Avoidance and Management of Pregnancy ........................................................................ 27 
8 
Adverse Events ............................................................................................................... 28 
8.1 
Definitions ....................................................................................................................... 28 
8.1.1 
Assessment of severity .................................................................................................... 29 
8.1.2 
Assessment of Causality .................................................................................................. 29 
8.2 
Collection and follow up of adverse events ................................................................ 30 
8.3 
Serious Adverse Events (SAE) ...................................................................................... 30 
8.4 
Liver Events and possible suicide related events ...................................................... 31 
9 
DATA HANDLING ................................................................................................................ 31 
9.1  
Recording of Data ........................................................................................................... 32 

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9.2  
Source Documentation and Study Records ................................................................ 32 
9.3 
 Data Management .......................................................................................................... 32 
9.4 
 Archiving and storage of data ...................................................................................... 32 
10  QUALITY CONTROL AND QUALITY ASSURANCE .......................................................... 33 
10.1 
Monitoring Arrangements .............................................................................................. 33 
10.2 
Quality Assurance .......................................................................................................... 33 
11 
ADMINISTRATIVE PROCEDURES ..................................................................................... 33 
11.1 
Ethics Approval .............................................................................................................. 33 
11.2 
Regulatory Notification .................................................................................................. 34 
11.3 
Insurance Provisions ..................................................................................................... 34 
11.4 
Publication Policy ........................................................................................................... 34 
11.5 
 Drug Accountability ...................................................................................................... 34 
11.6 
 Sample shipment and processing ............................................................................... 35 
REFERENCES: ............................................................................................................................. 35 
APPENDICES ............................................................................................................................... 37 
Appendix 1: 
Study flowchart ................................................................................................ 37 
Appendix 2: 
Collecting, processing, shipping & storing of blood & plasma .................. 39 
Appendix 3: Division of AIDS (DAIDS) Table for Grading the Severity of Adult and Pediatric 
Adverse Events ............................................................................................................................ 39 
Appendix 4:  Risk Assessment:  Dolutegravir (TIVICAY) ........................................................ 39 
Appendix 5: Liver Chemistry Stopping and Follow up Criteria .............................................. 41 
Appendix 6:Highly Effective Methods for Avoiding Pregnancy in Women of Child Bearing 
Potential* ...................................................................................................................................... 43 
 
 
 
 

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1. 
Background Information  
1.1 Introduction & background 
Metabolic changes and weight increase commonly occur early in ART. Weight gain observed in 
many patients shortly after ART initiation is thought to be due in part to a reduction in basal 
metabolic rate following suppression of plasma viremia, improved appetite, lower inflammatory 
cytokine effects  and a reduction in the rate of protein turnover [1, 2].  
Multiple studies in healthy volunteers have demonstrated acute onset of reversible insulin 
resistance, as assessed by euglycaemic hyperinsulinaemic clamp, inhibition of glucose 
transporters, in particular GLUT4, with  protease inhibitors (PIs) and difference in effects on insulin 
resistance and other metabolic parameters is seen as a key differentiator of agents within the PI 
class. Other antiretroviral drug classes have also been extensively tested. Clinical cohort data and 
data from clinical trials have reported an association between some nucleoside reverse 
transcriptase inhibitors (NRTIs) (stavudine, zidovudine, didanosine) and insulin resistance or 
diabetes mellitus. As mitochondrial dysfunction has been proposed as playing a role in the 
pathogenesis of diabetes mellitus, the association with drugs which may impact mitochondrial 
DNA and diabetes is not surprising. Tenofovir DF use does not cause insulin resistance when 
investigated with euglycemic clamp methodology [3]. 
A recent case report of acute onset diabetes mellitus after a switch to an Integrase Strand Transfer 
Inhibitor (INSTI)-containing regimen (raltegravir/abacavir/lamivudine) postulated effects on 
bioavailable magnesium could alter insulin signalling characteristics [4].   
The association between insulin resistance and body shape changes and weight gain during 
therapy has not been established. Insulin resistance triggered by some ARV drugs is apparently 
reversible, following switching to an agent not associated with insulin resistance. However, 
reduction in central adiposity has only been observed in highly selected subject, [5] weight gain 
in the setting of HIV may not be exclusively related to glucose and/ or fat handling, but may also 
relate to alterations in endocrine function, therefore we proposed study to investigate changes in 
insulin resistance, lipid metabolism and complete endocrine profile in subject exposed to 
dolutegravir to investigate the role all these different factors may potentially have in weight gain 
recently reported in clinical cohorts.  
1.2 Rationale 
In a retrospective analysis in the NA-ACCORD, 22% of those with normal BMI at the start of anti-
retroviral therapy (ART) became overweight after 3 years of ART, and 18% of those overweight at 
baseline becoming obese [1]. And a recent retrospective analysis by Menard et al [6] on patients 
stopping dolutegravir (DTG) due to adverse effects in the clinical setting, found that 7% had done 
so due to weight gain. Norwood and colleagues [7] has reported significantly increased weight 
gain in those switching to an INSTI-based regimen from an efavirenz-based (Atripla) based 
regimen. This weight gain was greatest among patients switching to DTG/ABC/3TC [7]. The 
ethology of weight gain in patients with undetectable plasma HIV RNA who change to another 
regimen and remain suppressed is unclear. An abrupt reduction in insulin sensitivity could 

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promote storage of excess circulating glucose and lipids in adipose tissue, but this is was 
speculative until now, therefor we propose the study of insulin sensitivity to address the 
underlining reasons for weight change in patients receiving dolutegravir based therapy.  
Other studies have used a range of ART for 28 days in healthy volunteers looking at similar 
metabolic, coagulation and inflammatory markers and pharmacokinetic [8-11]. Given DTG half-
life we wished to establish steady state to adequately examine the effect of a drug on weight 
gain, lipids and neuroendocrine markers. 28 days of DTG is Centre for Disease Control (CDC) 
recommended post exposure prophylaxis (PEP), therefore this duration of dosing is wildly used 
and recommended in HIV-negative individuals [12].  
 
HYPOTHESIS:  There is no difference in change in insulin resistance and multiple lipid 
parameters in healthy volunteers when treated with dolutegravir compared with baseline 
values.   
 
2. 
Trial Objectives  
2.1 
Primary Objective 
To quantify the difference in change in insulin sensitivity (determined by peripheral glucose 
uptake using a euglycaemic clamp) with the administration of dolutegravir (DTG) compared to no 
DTG for 28 days in HIV-seronegative healthy volunteers.  
2.2   
Secondary Objectives 
To investigate the effect of the dolutegravir on: 
1. Adipocytokines by assessing fasting adiponectin and leptin levels 
2. Fasting Ghrelin  
3. Pituitary hormones  
4. Lipid profile including lipid fractions 
5. Changes in indirect calorimetry 
6. Changes in food intake by food diaries  
7. Change in sleep parameters by sleep questionnaires  
8. Safety and tolerability of dolutegravir 
 
3. 
Trial Design 
3.1 
Endpoints  
Primary Endpoint 
▪ 
Change from baseline in total body glucose disposal by euglycaemic clamp method with 28 
days dolutegravir treatment. 

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CRF003 protocol version 4.0 20 Jan 2023 
 
Secondary Endpoints 
▪ 
Change from baseline in serum levels of fasting cholesterol and triglycerides 
▪ 
Changes in adipocytokines, ghrelin and pituitary  hormones  
▪ 
Changes in indirect calorimetry 
▪ 
Changes in food intake by food questionnaires  
▪ 
Changes in sleep quality by sleep questionnaires  
▪ 
Safety data including the number and severity of adverse events, as assessed by the medical 
dictionary for regulatory activities (MedDRA), from baseline to day 100  
 
 
3.2 
Study design 
This is a 72-day (excluding screening period and follow up phone call) randomized study with two 
arms.  The study will utilize a crossover design as described below.  
Subjects will be randomised to start on dolutegravir 50 mg once daily OR no treatment for the 
first 28 day dosing phase of the study, then will cross over to the alternative for the second dosing 
phase, following a 2 week washout period (equivalent to 5+ dolutegravir elimination half-lives). 
Subjects will be contacted approximately seven days after the final euglycaemic clamp visit for a 
telephone follow up. 
• 
Day -28 to 0:                    Screening 
• 
Day 1 (am):                      Euglycaemic clamp 1 
• 
Day 1 (pm) to 28:            Dolutegravir (n=8)/Control (n=8) 
• 
Day 28 (am):                    Euglycaemic clamp 2 
• 
Day 29 to 43:                   Wash out (14 days) 
• 
Day 44 to 72:                   Control (n=8)/Dolutegravir (n=8) 
• 
Day 72 (am):                    Euglycaemic clamp 3 
• 
Day 100                             Post study telephone contact   
3.3 
Randomisation  
After screening, eligible subjects satisfying the inclusion and exclusion criteria will be randomised 
1:1 to one of two arms:  
Group 1:  

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• Dolutegravir 50 mg once daily for the first 28 days of the study.  
• No treatment for the last 44 days of the study. 
Group 2:  
• No treatment for the first 43 days of the study.  
• Dolutegravir 50 mg once daily for the last 28 days of the study (day 44-72).  
3.4  
Benefit: Risk Assessment 
• 
Key identified and potential risks of DTG and the clamp procedure are summarized in 
Appendix 4. More detailed information about the known and expected risks and 
reasonably expected adverse events (AEs) of DTG may be found in the SmPC for Tivicay. 
(Appendix 4) 
4. 
Subject Population 
4.1  
Number of subjects and subject selection 
Subjects will be HIV-1 negative volunteers aged between 18 to 60 years inclusive.  The total 
number of subjects recruited will be 16 (8 per group) 
 
4.1.1 
Inclusion Criteria 
• 
Willing and able to provide informed consent 
• 
Cis-Male and Cis-Female healthy subjects without underlying conditions  
• 
Subjects must have documented negative HIV serology by ELISA and P24 antigen and not 
receiving anti-HIV pre-exposure  prophylaxis (PreP) 
• 
Subjects must be clinically well volunteers aged between 18 to 60 years with BMI <30 
kg/m2 but >18 kg/m2  
• 
Healthy, as determined by the investigator or medically qualified designee based on a 
medical evaluation, including medical history, physical examination, laboratory tests, and 
cardiac evaluation (including ECG)  
• 
Non-fasting blood glucose, total cholesterol and triglycerides within normal limits  
• 
Subjects should have complete blood count (FBC) with normal differential and platelet 
count (detail below of specified normal range, table 1) 
 
 
 
 
 

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Table 1 – Compete FBC with normal differential & platelets count ranges  
Test 
Male Normal Range 
Female Normal Range 
Haemoglobin (g/L) 
130-168 
114-150 
White blood cell count (x109/L) 
4.2-10.6 
4.2-11.2 
Neutrophil count (x109/L) 
2.0 - 7.1 
Lymphocyte count (x109/L) 
1.1 - 3.6 
Monocyte count (x109/L) 
0.2 - 0.9 
Eosinophil count (x109/L) 
0.0 - 0.5 
Basophil count (x109/L) 
0.0 - 0.2 
 
• 
A female, may be eligible to enter and participate in the study if she:  
o is of non-child-bearing potential defined as either post-menopausal (12 months 
of spontaneous amenorrhea and ≥45 years of age) or physically incapable of 
becoming pregnant with documented tubal ligation, hysterectomy or bilateral 
oophorectomy or, 
o is of child-bearing potential with a negative pregnancy test at both Screening and 
Day 1 and agrees to use one of the following methods of contraception to avoid 
pregnancy: 
o Complete abstinence from penile-vaginal intercourse. Abstinence is acceptable 
only as true abstinence when this is in line with the preferred and usual lifestyle 
of the participant; 
o Any intrauterine device with published data showing that the expected failure 
rate is <1% per year (not all intrauterine devices meet this criterion, see Appendix 
6] for an example listing of approved intrauterine devices); 
o Male partner sterilization confirmed prior to the female subject’s entry into the 
study, and this male is the sole partner for that subject; 
o Approved hormonal contraception (see Appendix 6] for a listing of examples of 
approved hormonal contraception)*; 
o Any other method with published data showing that the expected failure rate is 
<1% per year 
• 
Men who have partners who are women of childbearing potential (WOCBP – definition in 
Appendix 6) must be using an adequate method of contraception to avoid pregnancy in 
their partner throughout the study and for a period of at least 4 weeks after the study;   
 
Complete abstinence from penile-vaginal intercourse.  Abstinence is acceptable 
only as true abstinence when this is in line with the preferred and usual lifestyle 
of the patient; 
 
Double barrier method (male condom/spermicide, male condom/diaphragm, 
diaphragm/spermicide); 

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CRF003 protocol version 4.0 20 Jan 2023 
 
Any intrauterine device (IUD) with published data showing that the expected 
failure rate is <1% per year (not all IUDs meet this criterion, see Appendix 4 for an 
example listing of approved IUDs) plus male condom; 
 
Sterilisation confirmed prior to the subject’s entry into the study 
 
Approved hormonal contraception used by female partner (see protocol 
appendix 4 for a listing of examples of approved hormonal contraception) plus 
male condom; 
 
Any other method with published data showing that the expected failure rate is 
<1% per year and not containing hormones plus male condom. 
 
Any contraception method must be used consistently, in accordance with the 
approved product label and for at least four weeks after discontinuation of IMP 
(Appendix 6).  
Any contraception method must be used consistently, in accordance with the approved product 
label and for at least 28 days prior to the first dose of study medication and 4 weeks after 
discontinuing the study medication. 
4.1.2 
Exclusion Criteria  
• 
Subjects with a waist hip ratio > 0.97 or BMI > 30kg/m2 and BMI <18 kg/m2
 
will be excluded  
• 
Acute or chronic hepatitis B infection (determined by positive hepatitis B surface antigen 
result at the screening visit)  
• 
Acute or chronic hepatitis C infection (determined by positive hepatitis C antibody result 
at the screening visit)  
• 
Diabetes mellitus, other metabolic syndrome or disease process in the opinion of the 
investigator likely to cause marked disturbance in glucose and lipid homeostasis including 
hypertension. Subject with HbA1c >42 mmol/mol will be excluded. 
• 
History or presence of allergy to the dolutegravir  
• 
ALT or AST greater than or equal to 1.5 x Upper Limit of Normal (ULN) and total bilirubin 
greater than or equal to 1.5 x ULN excluded; 
• 
Pregnancy and breastfeeding women  
• 
Alcohol consumption >10 units/week  
• 
Clinically relevant drug use (positive urine drug screen) or history of alcohol or drug use 
considered by the Investigator to be sufficient to hinder compliance with treatment, 
follow-up procedures or evaluation of adverse events.  Smoking is permitted, but tobacco 
intake should remain consistent throughout the study. 
• 
 Unable to refrain from the use of prescription (e.g., dofetilide) or non-prescription drugs, 
including vitamins, herbal and dietary supplements (including St John’s wort) within 7 

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CRF003 protocol version 4.0 20 Jan 2023 
days (or 14 days if the drug is a potential enzyme inducer) or 5 half-lives prior to the 
baseline visit and throughout the study until the follow-up period, unless in the opinion 
of the Investigator the medication will not interfere with the study procedures or 
compromise participant safety.  
• 
This includes on-going therapy with any of the following   
- Metabolically active medications 
- Any lipid-lowering medication 
-Any testosterones treatments or supplements - Glucocorticoids including inhaled 
steroids except for ‘as necessary’ use 
- Beta-blockers  
- Thiazide diuretics and indapamide 
- Thyroid preparations  
- Psychotropic agents  
- Anabolic steroids  
- Megestrol acetate  
- Dofetilide (or pilsicainide) 
4.2  
Duration of Involvement  
Total duration of involvement in the study is up to 72 days, plus a screening visit within 28 days 
prior to day 1 and follow up telephone call approximately 28 days after the last euglycaemic 
clamp. 
4.3 
Withdrawal of Subjects and Discontinuation Criteria  
Subjects have the right to withdraw from the study at any time for any reason. The investigator 
also has the right to withdraw subjects from the study in the event of intercurrent illness, adverse 
events, protocol violations, administrative reasons or other reasons. An excessive rate of 
withdrawals can render the study impossible to interpret. Therefore, unnecessary withdrawal of 
subjects should be avoided. Should a subject decide to withdraw, all efforts will be made to 
complete and report the observations as thoroughly as possible. 
The investigator should contact the subject either by telephone or through a personal visit or a 
responsible relative must be contacted to determine as completely as possible the reason for the 
withdrawal. A complete final evaluation at the time of the patient's withdrawal should be made 
with an explanation of why the subject is withdrawing from the study.  If the reason for removal 
of a subject from the study is an adverse event or an abnormal laboratory test result, the principal 
specific event or test will be recorded on the Case Report Form. 
A subject will be considered to have withdrawn from the study if they do not complete all 
euglycaemic clamp procedures (day 1, 28 and day 72). 
Subjects may be replaced at the discretion of the investigator. 

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5. 
Treatment of subjects 
5.1. Study Treatment Regimen 
Group 1 
Subjects will receive dolutegravir 50 mg once daily from day 1 for 28 days. 
On the evening after the second clamp (day 28) they will cease treatment and will continue study 
activities for following 44 days.  
Group 2 
Subjects will receive no treatment from  day 1 for 43 days on the day 44 they will initiate 
dolutegravir 50 mg once daily for 28 days (until day 72). 
Dolutegravir will be dispensed by the Chelsea and Westminster NHS Trust pharmacy labelled as 
clinical trial supply. 
5.2 
Treatment Compliance 
Adherence to investigational product will be evaluated using pill counts of returned / unused 
investigational product. This assessment will be documented in the drug accountability record.  
6. 
Visit schedule & Procedures 
The schedule of assessments is summarised in Appendix 1. 
Screening Visit 
The screening visit should take place within 28 days prior to study day 1. 
Subjects will be provided with written information about the study in the form of a subject 
information sheet and will be allowed adequate time for questions and to consider the study 
before agreeing to participate.  It will be the responsibility of the investigator or co-investigator 
to obtain written informed consent prior to undertaking any procedures detailed in the protocol. 
The investigator or designee must provide adequate explanation of the aims, methods, objectives 
and potential hazards of the study.  It must also be explained to the subject that they are free to 
refuse or withdraw from the study for any reason without detriment to their future care or 
treatment. 
The following information and procedures will be recorded and performed as part of the screening 
assessments: 
• 
Written Informed Consent 
• 
Medical and medication history, including smoking and substance history 
• 
Demographics; gender, ethnic origin, age, height, weight, and body mass index (BMI) 
• 
Vital signs: temperature, blood pressure, heart rate, respiration rate, oxygen saturation 
• 
Physical examination including measurement of waist and hip size 

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CRF003 protocol version 4.0 20 Jan 2023 
• 
Haematology complete blood count (FBC) with differential and platelet count 
(haemoglobin, haematocrit, erythrocyte count, white blood count, differential white 
blood cell count, platelets)  
 
Pregnancy testing and last menstrual period (LMP) date in WOCBP subjects 
• 
Biochemistry safety labs assessment for standard clinical chemistry (glucose, creatinine, 
sodium, potassium, calcium, aspartate aminotransferase, alanine aminotransferase, 
bilirubin, CRP), including HbA1c 
• 
HIV antibody testing, hepatitis C (antibody) and B (surface antigen) screening 
• 
ECG 
• 
Urinary analysis (pH, specific gravity, leucocytes, nitrite, protein, glucose, ketones, 
urobilinogen, bilirubin, erythrocytes)   and urine drug screen 
 
Baseline  
Subjects who meet all the inclusion and exclusion criteria and wish to take part in the study will 
be admitted to the unit on the morning of day 1.   Subjects will remain in the unit for 
approximately four to six hours.  
The following study assessments will be performed (subjects must have fasted for at least eight 
hours): 
• 
Adverse events 
• 
Concomitant medication  
• 
Vital signs (during the clamp procedure vital signs will occur every 30 minutes)  
• 
Height/ weight/ BMI  
• 
Measurement of waist and hip size 
• 
Haematology: complete blood count (FBC) with differential and platelet count 
(haemoglobin, haematocrit, erythrocyte count, white blood count, differential white 
blood cell count, platelets) 
• 
Biochemistry safety labs assessment for standard clinical chemistry (fasting glucose, 
creatinine, sodium, potassium, calcium, aspartate aminotransferase, alanine 
aminotransferase, bilirubin), including thyroid hormones (TSH, ft4), lipids 
(triglycerides, cholesterol, cholesterol HDL & LDL Sub-fractions) CRP and Hb1aC   
• 
Urinary analysis (pH, specific gravity, leucocytes, nitrite, protein, glucose, ketones, 
urobilinogen, bilirubin, erythrocytes)   and urine drug screen 
• 
Pregnancy testing and LMP date in WOCBP subjects  
• 
Adiponectin, leptin, ghrelin, testosterone 
• 
Pituitary hormone function tests 

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CRF003 protocol version 4.0 20 Jan 2023 
• 
Dietary Questionnaire  
• 
Sleep Questionnaires (PSQI) 
• 
Euglycaemic clamp 1 
• 
Insulin will be titrated 3 times during euglycaemic clamp procedure according to approved 
and verified formula sheet. 
• 
ECG monitoring at 30 minute intervals during the clamp procedure 
• 
Indirect calorimetry (all participants) 
 
Prior to baseline visit subjects will be randomly assigned to Group 1 or Group 2: 
 
Group 1:  
 
• Dolutegravir 50 mg once daily for the first 28 days of the study.  
 
• No treatment for the last 44 days of the study. 
Subject will be dispensed study medication (Dolutegravir 50 mg) and instructed to self-administer 
the study drugs at home from the morning of day 2 until the morning prior to the next euglycaemic 
clamp day (Day 27). 
 
 
Group 2:  
 
No treatment for the first 43 days of the study.  
 
Dolutegravir 50 mg once daily for the last 28 days of the study (day 44-72).  
Subject will be dispensed study medication (Dolutegravir 50 mg) and instructed to self-administer 
the study drugs at home from the morning of day 45 until the morning prior to the next 
euglycaemic clamp day (Day 71). 
 
Day 28  
Subjects will be admitted to the unit in the morning and remain on the unit for approximately four 
to six hours. Group 1 subjects will take their last dose of study medication in the unit in the 
morning of the visit.  
The following study assessments will be performed (subjects will have fasted for at least eight 
hours): 
• 
Adverse events 
• 
Concomitant medication  
• 
Vital signs (during the clamp procedure vital signs will occur every 30 minutes)  
• 
Height/ weight/ BMI  
• 
Measurement of waist and hip size 

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CRF003 protocol version 4.0 20 Jan 2023 
• 
Haematology: complete blood count (FBC) with differential and platelet count 
(haemoglobin, haematocrit, erythrocyte count, white blood count, differential white 
blood cell count, platelets) 
• 
Biochemistry safety labs assessment for standard clinical chemistry (fasting 
glucose,creatinine, sodium, potassium, calcium, aspartate aminotransferase, alanine 
aminotransferase, bilirubin), including thyroid hormones (TSH, ft4), lipids 
(triglycerides, cholesterol, cholesterol HDL & LDL Sub-fractions) CRP and HbA1c   
• 
Urinary analysis (pH, specific gravity, leucocytes, nitrite, protein, glucose, ketones, 
urobilinogen, bilirubin, erythrocytes)   and urine drug screen 
• 
Pregnancy testing and LMP date in WOCBP subjects  
• 
Adiponectin, leptin, ghrelin, testosterone  
• 
Pituitary hormone function tests 
• 
Dietary Questionnaire  
• 
Sleep Questionnaires (PSQI) 
• 
Euglycaemic clamp 2 
• 
Insulin will be titrated 3 times during euglycaemic clamp procedure according to approved 
and verified formula sheet. 
• 
ECG monitoring at 30 minute intervals during the clamp procedure 
• 
Indirect calorimetry (Group 1) 
 
Day 29 - Day 43 
Wash out Period  
 
Day 44 
Subjects will be admitted to the unit in the morning and remain on the unit for approximately one 
to two hours. Subjects randomized to Group 2 will take their first dose of study medication in the 
unit in the morning of the visit.  
The following study assessments will be performed (subjects must have fasted for at least 8 
hours): 
• 
Adverse events 
• 
Concomitant medication  
• 
Vital signs: temperature, blood pressure, heart rate, respiration rate, oxygen saturation 
• 
Height/ weight/ BMI  

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CRF003 protocol version 4.0 20 Jan 2023 
• 
Measurement of waist and hip size 
• 
Haematology: complete blood count (FBC) with differential and platelet count 
(haemoglobin, haematocrit, erythrocyte count, white blood count, differential white 
blood cell count, platelets) 
• 
Biochemistry safety labs assessment for standard clinical chemistry (fasting glucose, 
creatinine, sodium, potassium, calcium, aspartate aminotransferase, alanine 
aminotransferase, bilirubin), including thyroid hormones (TSH, ft4), lipids 
(triglycerides, cholesterol, cholesterol HDL & LDL Sub-fractions) CRP and HbA1c   
• 
Urinary analysis (pH, specific gravity, leucocytes, nitrite, protein, glucose, ketones, 
urobilinogen, bilirubin, erythrocytes)   and urine drug screen 
• 
Pregnancy testing and LMP date in WOCBP subjects  
• 
Adiponectin, leptin, ghrelin, testosterone 
• 
Pituitary hormone function tests 
• 
Dietary Questionnaire  
• 
Sleep Questionnaires (PSQI) 
Day 72  
Subjects will be admitted to the unit in the morning and remain on the unit for approximately four 
to six hours. Subjects will take their last dose of study medication (Group 2) in the unit in the 
morning of the visit.  
The following study assessments will be performed (subjects must have fasted for at least 8 
hours): 
• 
Adverse events 
• 
Concomitant medication  
• 
Height/ weight/ BMI  
• 
Vital signs (during the clamp procedure vital signs will occur every 30 minutes)  
• 
Measurement of waist and hip size 
• 
Haematology: complete blood count (FBC) with differential and platelet count 
(haemoglobin, haematocrit, erythrocyte count, white blood count, differential white 
blood cell count, platelets) 
• 
Biochemistry safety labs assessment for standard clinical chemistry (fasting glucose, 
creatinine, sodium, potassium, calcium, aspartate aminotransferase, alanine 
aminotransferase, bilirubin), including thyroid hormones (TSH, ft4), lipids 
(triglycerides, cholesterol, cholesterol HDL & LDL Sub-fractions) CRP and Hb1aC   

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CRF003 protocol version 4.0 20 Jan 2023 
• 
Urinary analysis (pH, specific gravity, leucocytes, nitrite, protein, glucose, ketones, 
urobilinogen, bilirubin, erythrocytes)   and urine drug screen 
• 
Pregnancy testing and LMP date in WOCBP subjects  
• 
Adiponectin, leptin, ghrelin, testosteron  
• 
Pituitary hormone function tests 
• 
Dietary Questionnaire 
• 
Sleep Questionnaires (PSQI) 
• 
Euglycaemic clamp 3 
• 
Insulin will be titrated 3 times during euglycaemic clamp procedure according to approved 
and verified formula sheet.  
• 
ECG monitoring at 30 minute intervals during the clamp procedure 
• 
Indirect calorimetry (Group 2) 
 
Follow up (Day 100) 
The subject will be contacted by telephone approximately 28 days after the final euglycaemic 
clamp day to check whether they have experienced any adverse events. This follow up phone call 
should be documented in the subject’s notes. WOCBP should take a home pregnancy test. 
Early termination visit 
If a subject withdraws or is discontinued from the study the subject should attend if at all possible 
for a discontinuation visit and undergo the following procedures: 
• 
Haematology: complete blood count (FBC) with differential and platelet count 
(haemoglobin, haematocrit, erythrocyte count, white blood count, differential white 
blood cell count, platelets) 
• 
Biochemistry safety labs assessment for standard clinical chemistry (fasting 
glucose,creatinine, sodium, potassium, calcium, aspartate aminotransferase, alanine 
aminotransferase, bilirubin), including thyroid hormones (TSH, ft4), lipids 
(triglycerides, cholesterol, cholesterol HDL & LDL Sub-fractions) CRP and Hb1aC   
• 
Symptom-directed physical examination (if any adverse event experienced)  
6.1 
Euglycaemic Clamp 
Following completion of the procedures mentioned above on days 1, 28 and 72 a euglycaemic 
clamp study will be performed to determine the extent of total and non-oxidative glucose 
disposal. After an overnight fast of at least 8 hours, the subject will be admitted to the Clinical 
Research Facility.  This procedure will take approximately 3 hours. A primed/ continuous 
intravenous infusion of insulin, through an intravenous cannula, will commence in the morning at 

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CRF003 protocol version 4.0 20 Jan 2023 
an infusion rate dictated by the verified formula sheet, maintaining a mid-physiological level of 
hyperinsulinaemia for 180 minutes . Whole blood glucose will be assayed from a separate 
intraveneous cannula every 5 minutes. A negative feedback formula will be used to adjust the rate 
of infusion of 20 % dextrose in order to maintain euglycaemia at 4.5 mmol.L-1. Whole blood 
potassium and haematocrit is measured at 30 minute intervals during the clamp procedure with 
electrocardiographic monitoring half hourly in order to monitor for changes towards 
hypokalaemia.  
At 180 min, the insulin and glucose infusion is stopped immediately, and the subject fed. A final 
blood sugar, potassium and haematocrit level, and vital signs check will be performed before the 
subject is discharged from the unit.  
The visit window for the euglycaemic clamp is up to plus or minus two days. All other visits should 
be adjusted accordingly. 
 
6.2 Indirect Calorimetry  
Indirect calorimetry by ventilated hood expires gas analysis will be used to determine energy 
expenditure during the course of the visit. Indirect calorimetry is the method by which the type 
and rate of substrate utilization, and energy metabolism are estimated in vivo starting from gas 
exchange measurements (carbon dioxide production and oxygen consumption during rest and 
steady-state exercise). Indirect calorimetry with hood canopy and dilution pump will be 
performed. Duration of indirect calorimetry is 30-40 minutes.  
7 
Statistical Analysis 
Statistical analysis will be carried out after all subjects complete the study. All patients who have 
at least one dose of the trial medication, whether withdrawn prematurely or not, will be 
included in the safety analysis. 
All safety data and adverse events will be summarised. 
7.1 
Sample size 
No formal power calculations have been performed however, based on a treatment study in HIV-
negative healthy volunteers over 4 weeks therapy [2, 9-11]; a power calculation can be 
determined by using the change in insulin-mediated glucose uptake after four weeks of therapy 
as the primary end point. 
The study with eight [5] subjects per sequence will achieve 80% power to show equivalency in 
insulin-mediated glucose disposal under DTG treatment between Baseline and Week 4, assuming 
a standard deviation of difference between baseline and week 4 is 1.4 mg/kg x min per 
microUnit/mL insulin [1], the insulin-mediated glucose disposal equivalence boundary will range 
between −1.1 mg/kg × min per microUnit/mL to 1.1 mg/kg × min per microUnit/mL.  
The primary endpoint of this study is the change from baseline in insulin sensitivity by euglycaemic 
clamp method with 28 days treatment. 

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CRF003 protocol version 4.0 20 Jan 2023 
The secondary endpoints include:  
• 
Change from baseline in serum levels of fasting cholesterol, triglycerides 
• 
Changes from baseline in serum adipocytokines, ghrelin and pituitary hormones  
• 
Changes from baseline in indirect calorimetry  
7.2   Criteria for evaluation  
All subjects who complete day 1, 28, 44 and 72 assessments (including all euglycaemic clamps) 
will form part of the evaluable cohort.  
7.3 
Statistical Methods 
Baseline demographic characteristics will be summarized using the descriptive statistics by 
treatment arms. Where quantitative data are hyper geometrically distributed median with inter-
quartile ranges will be presented while Gaussian normal data will be summarized using the  mean 
and standard deviation while qualitative data will be summarized using number of subjects with 
percentages.  
The primary efficacy endpoint will be evaluated using analysis of variance (ANOVA) or Analysis of 
covariance (ANCOVA) fitting models for the crossover experiment, including treatment, period, 
and sequence as fixed effects, and subject within sequence as a random effect in the model. Least 
square mean difference between dolutegravir and no treatment period will be obtained from the 
ANOVA, or ANCOVA adjusting for potential confounding factors and the associated two-sided 95% 
confidence intervals will be constructed. It will be concluded that dolutegravir has equivalent 
impact as no treatment on insulin-mediated glucose disposal if the 90% confidence interval for 
the mean difference between the two treatments is within −1.1 mg/kg × min per microunit/mL to 
1.1 mg/kg × min per microunit/mL boundary.  
All data analyses will be performed using STATA statistical software (version 17). All p-values 
presented will be two tailed. 
All patients who attend the baseline visit whether withdrawn prematurely or not will be included 
in the safety analysis. All safety data and adverse events will be summarised. Adverse events will 
be mapped using medical dictionary for regulatory activities (MedDRA) (the latest available 
version). The incidence of adverse event will be tabulated by system organ class, preferred term, 
and treatment. Additional summaries of adverse events will be displayed by highest grade, 
investigator’s assessment of relationship to study drug, and effect on study drug discontinuation.  
Statistical methods and power calculations for the study have been previously published by 
multiple different groups [3, 14-16].   
Exploratory analyses will look at the effects of age, baseline BMI, sex and race on treatment 
effects. 

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7.3.1 Efficacy parameters 
There will be no assessment for efficacy over the course of the study as the subjects are HIV-1 
seronegative volunteers. 
7.3.1 Safety parameters 
All patients who have at least one dose of the trial medication, whether withdrawn prematurely 
or not, will be included in the safety analysis. 
All safety data and adverse events will be summarised.  
7.4 
Criteria for termination of the trial 
The Sponsor or Investigator may terminate either part of, or the entire study for safety or 
administrative reasons.  A written statement fully documenting the reasons for such a 
termination will be provided to the Ethics Committee and the Regulatory Authority as 
appropriate. 
7.5 Toxicity Management 
In the event of a discontinuation of a DTG for suspected drug induced liver injury, other clinically 
significant liver chemistry elevations, severe skin reaction or hypersensitivity reaction, subjects 
should not be re-challenged with a DTG-containing product due to the risk of a recurrent reaction. 
These subjects should be withdrawn from study. 
• 
Allergic reaction  
Subjects with Grade >1 or higher allergic reactions that are considered to be possibly or probably 
related to the investigational product should permanently discontinue the investigational product 
and the subject should be withdrawn from the study. Subjects should be treated as clinically 
appropriate and followed until resolution of the adverse event. 
• 
Rash 
Participants should be instructed to contact the Investigator as soon as possible if they develop a 
rash while on study. Mild to moderate rash is an expected adverse reaction for DTG-containing 
antiretroviral therapy.  
Subjects should permanently discontinue investigational product (and all other concurrent 
medication(s) suspected in the Investigators causality assessment) for a rash Grade 1 or higher, 
and the subject should be withdrawn from the study. Subjects should be treated as clinically 
appropriate and followed until resolution of the adverse event. 

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CRF003 protocol version 4.0 20 Jan 2023 
The rash and any associated symptoms should be reported as adverse events and appropriate 
toxicity ratings will be used to grade the events (based on DAIDS [Division of AIDS] toxicity 
grading). 
• 
Liver Chemistry Stopping and Follow up Criteria Summarized in Appendix 5. 
• 
Suicidal Risk Monitoring 
Subjects will be monitored appropriately and observed closely for suicidal ideation and behaviour, 
or any other unusual changes in behaviour. Mental health consultation will be considered or 
referral for subjects who experience signs of suicidal ideation or behaviour. 
Subjects with evidence of suicidal ideation or behaviour changes should permanently discontinue 
the investigational product and the subject should be withdrawn from the study.  
If any subject experiences a possible suicidality-related adverse event (PSRAE) while participating 
in this study that will be considered to meet ICH E2A (ICH E2A, 1994) definitions for seriousness, 
we will collect information using a PSRAE case report form in addition to reporting the event on a 
serious adverse event case report form. A PSRAE may include, but is not limited to, an event that 
involves suicidal ideation, a preparatory act toward imminent suicidal behaviour, a suicide 
attempt, or a completed suicide.  The investigator will exercise his or her medical and scientific 
judgment in deciding whether an event is possibly suicide-related. PSRAE forms should be 
completed and reported to Chelsea and Westminster NHS Trust R&D within one week of the 
investigator diagnosing a possible suicidality-related serious adverse event. 
7.6 Avoidance and Management of Pregnancy 
Due to the potential risk from neural tube defect associated with DTG treatment during the first 
trimester of pregnancy, study staff must review the following information with all female 
participants of reproductive potential: 
 
Participants should be informed that in an early analysis of one observational study, 
women who were taking DTG when they became pregnant had an increased risk of having 
babies with serious brain and spine defects. These defects happen early in pregnancy, 
before many women even know they are pregnant. 
 
Women of reproductive potential should be counselled on the importance of avoiding 
pregnancy, safer sexual practices and the proper use of their chosen contraceptive 
methods in accordance with the applicable contraceptive product label, or, for non-
product methods, as determined by the investigator. 

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CRF003 protocol version 4.0 20 Jan 2023 
 
The need for the participant’s chosen contraceptive method to be used for an adequate 
time period before dosing with the DTG is initiated. 
 
The need to continue the use of contraception throughout the treatment- and post-
treatment- periods with DTG, until it is predicted that a clinically insignificant amount of 
DTG is present in the participant (i.e. 30 days or one month after stopping DTG). 
 
Females of reproductive potential should be reminded about the importance of 
pregnancy avoidance and of adherence to contraception requirements at every study 
visit. 
 
Additionally, site staff should document the participant’s chosen contraception methods in her 
medical records and study records/eCRF. 
Any subject who becomes pregnant during the study should be immediately withdrawn from DTG, 
to eliminate further exposure to the embryo/foetus, and withdrawn from the study.  Any 
pregnancy that occurs during study participation must be reported using the Pregnancy CRF 
(Appendix 6).  To ensure subject safety, each pregnancy must be reported to the PI and the 
nominated person at the sponsor Chelsea and Westminster NHS Trust  research and development 
(R&D) office (as set by the sponsor standard operating procedures (SOP)), within 24 hours of 
learning of its occurrence. The pregnancy should also be reported to ViiV Healthcare in accordance 
with the terms of the contract. The pregnancy must be followed up to determine outcome 
(including premature termination) and status of mother and child, which must also be reported 
to the PI and the nominated person at the sponsor R&D office, using a the Pregnancy Follow-up 
CRF (Appendix 6). The pregnancy outcome should also be reported to ViiV Healthcare within one 
week of awareness in accordance with the contract. Pregnancy complications and elective 
terminations for medical reasons must be reported as an adverse event or serious adverse event.  
Spontaneous abortions must be reported as a serious adverse event. 
Any serious adverse event occurring in association with a pregnancy brought to the investigator’s 
attention after the subject has completed the study and considered by the investigator as possibly 
related to DTG, must be promptly reported to the PI and the nominated person at the sponsor 
R&D office and ViiV Healthcare.  
8 
Adverse Events 
8.1 
Definitions 
An adverse event (AE) is any untoward medical occurrence in a subject administered a 
pharmaceutical product and which does not necessarily have a causal relationship with the study 

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CRF003 protocol version 4.0 20 Jan 2023 
treatments. An AE can therefore be any unfavourable and unintended sign (including an abnormal 
laboratory finding), symptom, or disease temporally associated with the use of a medicinal 
(investigational) product, whether or not considered related to the medicinal (investigational) 
product. 
Adverse events observed by the investigator, or reported by the subject, and any remedial action 
taken, will be recorded in the subject’s CRF and should be verifiable in the subject’s notes 
throughout the study.  The nature of each event, time of onset after drug administration, duration 
and severity will be documented together with the Investigator’s opinion of the causal 
relationship to the treatment (unrelated, unlikely, possible, probable, and definite). 
All subjects experiencing adverse events, whether considered associated with the use of the study 
medication or not, must be monitored until the symptoms subside and any clinically relevant 
changes in laboratory values have returned to baseline, or until there is a satisfactory explanation 
for the changes observed. 
Procedures such as surgery should not be reported as adverse events.  However, the medical 
condition for which the procedure was performed should be reported if it meets the definition of 
an adverse event.  For example, an acute appendicitis that begins during the adverse event 
reporting period should be reported as the adverse event and the resulting appendectomy noted 
on the CRF. 
Planned procedures such as surgery planned prior to the subject’s enrolment into the study need 
not be reported as adverse events if these are documented as planned at the screening visit. 
Clinically significant changes in physical examination and blood safety profiles should also be 
recorded as adverse events. 
8.1.1 
Assessment of severity 
Severity should be recorded and graded according to the current version of the Division of AIDS 
(DAIDS) Table for Grading the Severity of Adult and Paediatric Adverse Events (Appendix 3).  
Note: There is a distinction between the gravity and the intensity of an adverse event. Severe is a 
measure of intensity; thus, a severe reaction is not necessarily a serious adverse event. For 
example, a headache may be severe in intensity but would not be classified as serious unless it 
met one of the criteria for serious events. 
8.1.2 
Assessment of Causality 
The relationship to study drug of each adverse event will be assessed using the following 
definitions: 
DEFINITE: 
Distinct temporal relationship with drug treatment. Known reaction to agent or 
chemical group, or predicted by known pharmacology. Event cannot be explained 
by subject’s clinical state or other factors. 

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CRF003 protocol version 4.0 20 Jan 2023 
PROBABLE: 
Reasonable temporal relationship with drug treatment. Likely to be known 
reaction to agent or chemical group, or predicted by known pharmacology. Event 
cannot easily be explained by subject’s clinical state or other factors. 
POSSIBLE: 
Reasonable temporal relationship with drug treatment. Event could be explained 
by subject’s clinical state or other factors. 
UNLIKELY: 
Poor temporal relationship with drug treatment. Event easily explained by 
subject’s clinical state or other factors. 
UNRELATED: 
This is an event which occurs before dosing. Event or intercurrent illness is due 
wholly to factors other than drug treatment. 
8.2 
Collection and follow up of adverse events 
All adverse events, however minor, will be documented in the CRF whether or not the investigator 
concludes the event to be related to drug treatment. 
The adverse event reporting period will be from the time that the subject gives informed consent 
until the subjects final study visit.  In addition, any untoward event that may occur subsequent to 
the reporting period that the Investigator assesses as possibly, probably or definitely related to 
the study drug medication should also be reported as an adverse event (AE). 
Adverse events may be directly observed, reported spontaneously by the subject or by 
questioning the subject at each study visit. 
All AE should be followed up until they are resolved or the subject’s participation in the study ends 
(i.e. until the final CRF is completed for that subject).  In addition, all serious and non-serious 
adverse events assessed by the Investigator as possibly related to the investigational medication 
should continue to be followed even after the subject’s participation in the study is over.  Such 
events should be followed until resolution, or until no further change can reasonably be expected.  
Deaths occurring more than 30 days after the final dose, which are considered to be unrelated to 
the study medication, should not be reported as SAE. 
8.3 
Serious Adverse Events (SAE) 
A serious adverse event is any untoward medical occurrence that at any dose: 
i) 
Results in death 
ii) 
Is life threatening 
iii) 
Requires in patient hospitalisation or prolongation of existing hospitalisation 
iv) 
Results in persistent or significant disability/ incapacity 
v) 
Is a congenital anomaly/ birth defect. 
vi) 
Medical or scientific judgment should be exercised in deciding whether reporting 
is appropriate in other situations, such as important medical events that may not 
be immediately life-threatening or result in death or hospitalisation but may 

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CRF003 protocol version 4.0 20 Jan 2023 
jeopardise the subject or may require medical or surgical intervention to prevent 
one of the other outcomes listed in the above definition.  These should also be 
considered serious.  Examples of such events are invasive or malignant cancers, 
intensive treatment in an emergency room or at home for allergic bronchospasm, 
blood dyscrasias or convulsions that do not result in hospitalisation, or 
development of drug dependency or drug abuse. 
vii) 
All events of possible drug-induced liver injury with hyperbilirubinemia defined 
as ALT 3xULN and bilirubin 2xULN (>35% direct). 
 
NOTE: bilirubin fractionation is performed if testing is available. If unavailable, the 
presence of detectable urinary bilirubin on a dipstick (indicating direct bilirubin 
elevations and suggesting liver injury) should be recorded. If testing is unavailable 
and a subject meets the criterion of total bilirubin 2xULN, then the event is still 
reported as a serious adverse event. If INR is obtained, include values on the 
serious adverse event form. INR elevations >1.5 suggest severe liver injury. 
 
The SAE should be reported immediately to the Principal Investigator (within 24 hours of a 
member of the study team becoming aware of the event).  SAEs, and Suspected Unexpected 
Serious Adverse Reactions (SUSARS) and pregnancies must be reported to the nominated person 
at the sponsor research and development office as set by the sponsor standard operating 
procedures (SOPs) within 24 hours of awareness of the event. All SAEs, SUSARs and pregnancies 
should also be notified to ViiV Healthcare, regardless of causality or expectedness, per the terms 
of the contract. The SAE should be reported to VIIV Healthcare within 24 hours of a member of 
the study team becoming aware of the event. A SAE form should be completed and an assessment 
of whether the SAE is a Suspected Unexpected Serious Adverse Reaction (SUSAR) conducted by 
the Principal Investigator. 
Section 4.8 in the Summary of Product Characteristics (SmPC) represent the Reference Safety 
Information to determine expectedness of serious adverse events for expedited reporting. 
The Principal Investigator is responsible for determining whether the SAE is a Suspected 
Unexpected Serious Adverse Reaction (SUSAR) and for reporting this in accordance with Good 
Clinical Practice (GCP) and applicable regulatory requirements. 
8.4 
Liver Events and possible suicide related events 
Additional reporting for Liver Events and Possible Suicide Related Events are detailed in Appendix 
5 and Section 7.5, respectfully. 
 
9 
Data handling 
 

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CRF003 protocol version 4.0 20 Jan 2023 
The results from the data collected during the study will be recorded in the Subjects Case Report 
Form (CRF).  In order to maintain confidentiality, the subject will be identified only by subject 
number and initials.  
9.1  
Recording of Data 
All data collected during the study will be recorded in an individual, subject specific CRF.  All CRF 
should be completed legibly in black ink by an appropriate member of the study team who should 
be identified and agreed by the Principal Investigator before the start of the study.  A delegation 
of responsibility signature log identifying personnel who can enter data and/or sign off a CRF will 
be maintained by the Principal Investigator.   
Corrections to the data on the CRF will only be made by drawing a single line through the incorrect 
data (so as not to obscure the original entry) and inserting the correct data next to the original 
entry.  The incorrect data must never be obliterated using correction fluid (e.g. Snopake® or 
Tippex®).  Each correction will be initialised and dated by the person making the correction. 
CRF should be kept current to enable the study monitor to review the subject status throughout 
the course of the study.  CRF will be completed, reviewed and signed off by the Investigator within 
two weeks of the last subject visit. 
9.2  
Source Documentation and Study Records 
The subject’s number and date of entry into the study, along with a study identifier, should be 
recorded in the subject’s study records.  The following should also be recorded in the study 
records; confirmation of written and oral consent, the subject’s clinical status, date of every study 
visit, date study medication was started and stopped, concomitant medications, copies of all 
relevant reports and laboratory tests, comments on results and reference to any adverse events. 
Euglycaemic clamp data will not be entered into the subject notes and the formula spreadsheet 
for this procedure will be considered to be source. 
9.3 
 Data Management 
Data will be entered into a study specific database by designated staff on a regular basis from 
completed Case Record Forms (CRF).  Data will be entered on a regular basis to ensure it is up to 
date. The database will be kept by SMART-TRIAL. Access to the database will be given to 
authorised personnel only and a log of authorised personnel will be stored in the trial master file. 
9.4 
 Archiving and storage of data 
Following completion of the study, subject records, CRF and other study documentation will be 
retained by the Investigator in accordance with Good Clinical Practice (GCP) and applicable 
regulatory requirements. 
 

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CRF003 protocol version 4.0 20 Jan 2023 
10  
Quality Control and Quality Assurance 
10.1 
Monitoring Arrangements 
The purpose of monitoring is to verify the rights and well-being of human subjects are protected; 
that trial data is accurate, complete and verifiable with source data; that the trial is conducted in 
compliance with the protocol, GCP and the applicable regulatory requirements. 
A monitor will conduct regular site visits for the purpose of monitoring various aspects of the 
study.  The Investigator must agree to allow the study monitor and authorised representatives of 
the sponsor to inspect all CRF and corresponding source documents, e.g. original medical records, 
subject records and laboratory raw data, access to the clinical supplies, dispensing and storage 
areas and agree to assist with their activities if requested.  The Investigator should provide 
adequate time and space for monitoring visits. 
The monitor will query any missing or spurious data with the Investigator, which should be 
resolved in a timely manner.  A monitoring log will be maintained recording each visit, the reason 
for the visit, the records reviewed and any findings. 
 
 10.2 Quality Assurance 
For the purpose of compliance with GCP and regulatory agency guidelines, it may be necessary 
for sponsor authorised Quality Assurance personnel and/or authorised personnel from an 
external regulatory agency to conduct an audit/inspection of an Investigational site.  The purpose 
of an audit is to assess the quality of data with regard to accuracy, adequacy and consistency, and 
to assure that studies are in accordance with Good Clinical Practices, and Regulatory Agency 
guidelines.  Having the highest quality data from studies is an essential aspect of drug 
development. 
The Investigator will be given sufficient notice to prepare for such visits, which are planned to take 
usually between one and two days and may be conducted at any stage during the study.  The 
audit will involve the review of all study related documentation, which is required by GCP to be 
maintained by site, review of drug storage, dispensing and return, review of all study related 
supplies and review of source documents against the CRF to assure the adequacy and accuracy of 
the information which has been recorded, including the verification of any AE which have 
occurred. 
11 
Administrative procedures  
11.1 
Ethics Approval 
 
The study protocol, subject information and consent form, available safety information, subject 
recruitment procedures, information about payments and compensation available to the subjects 
and documentation evidencing the Investigator’s qualifications should be submitted to the Ethics 
Committee for ethical review and approval according to local regulations, prior to the study start. 

34 
CRF003 protocol version 4.0 20 Jan 2023 
Any changes, which may need to be made, will be submitted in the form of numbered and dated 
protocol amendments in accordance with local regulations.  
11.2 
Regulatory Notification 
 
As required by local regulations, approval of the appropriate regulatory bodies will be obtained, 
prior to study initiation. 
11.3 
Insurance Provisions  
 
The Sponsor will take out appropriate insurance cover for this trial. 
11.4 
Publication Policy 
 
A whole or part of this study results will be communicated, orally presented, and/or published in 
appropriate scientific journals. Full anonymity of subject’s details will be maintained throughout. 
Subjects wanting to see the results of the trial can request a copy of the article from the 
investigators once it has been published. 
11.5 
 Drug Accountability 
The investigator will ensure that the investigational product will only be used in accordance with 
the protocol. Drug supplies will be kept in a secure, limited access storage area under the 
recommended storage conditions. 
Dolutegravir 50mg will be provided by ViiV Healthcare.  A drug accountability log will be kept with 
the investigational supplies for reconciliation purposes.  This should be used to record the 
identification of the subject to whom the investigational product was dispensed, the date and 
quantity of investigational product dispensed and the quantity of investigational product 
unused/returned by the subject. This will be verified by the study monitor. 
At the end of the study partially used or empty containers may be destroyed by pharmacy (in 
accordance with their usual procedures) after the study monitor has completed a final inventory.  
Due to the nature of the study and the need to dose subjects outside of normal working hours, 
pharmacy may need to prepare and dispense investigational medicinal product to the unit in 
advance of baseline visit. A process is in place to retrieve investigational medicinal product for 
those subjects who are withdrawn after medication has been dispensed to the unit and prior to 
dosing.  
Spare investigational medicinal product will be dispensed for replacement subjects in the event 
of withdrawals.  
Due to the nature of the study and intensive euglycaemic days occurring outside normal working 
hour’s pharmacy will dispense spare investigational medicinal product (IMP) to the unit. This 
IMP should only be used in the event of a subject failing to bring in their IMP. It is the 
responsibility of the investigator to release the IMP and to ensure that pharmacy is informed in 
order to document which subjects received the IMP.  Any IMP not required will be returned to 
pharmacy immediately. 
 

35 
CRF003 protocol version 4.0 20 Jan 2023 
A specific risk assessment for the concomitant use of a Covid-19 vaccine and Dolutegravir within 
the current protocol was performed. The outcome is that a Covid-19 vaccine given to a trial 
subject enrolled into the study is a simple concomitant medication with no interaction that 
requires advice on timing of the vaccine or other aspects.  
 
11.6 
 Sample shipment and processing 
Biochemistry, haematology, virology and immunology samples will be analysed by the local 
hospital lab according the local procedures. 
Pathology Laboratory 
Hammersmith Hospital NHS Trust 
Chelsea & Westminster Hospital 
369 Fulham Road 
London SW10 9NH 
 
 
References:  
1. 
Koethe JR, Jenkins CA, Lau B, Shepherd BE, et al. North American AIDS Cohort 
Collaboration on Research and Design (NA-ACCORD). Rising Obesity Prevalence and 
Weight Gain Among Adults Starting Antiretroviral Therapy in the United States and 
Canada. AIDS Res Hum Retroviruses. 2016; 32(1):50-8.  
2. 
Taramasso L, Ricci E, Menzaghi B, et al; CISAI Study Group. Weight Gain: A Possible Side 
Effect of All Antiretrovirals. Open Forum Infect Dis. 2017;  4(4):ofx239.  
3. 
Randell PA, Jackson AG, Zhong L, Yale K, Moyle GJ. The effect of tenofovir disoproxil 
fumarate on whole-body insulin sensitivity, lipids and adipokines in healthy 
volunteers.Antivir Ther. 2010; 15(2):227-33.  
4.           Fong PS, Flynn DM, Evans CD, Korthuis PT. Integrase strand transfer inhibitor-associated 
diabetes mellitus: A case report. Int J STD AIDS. 2017; 28(6):626-28.  
5. 
Moyle GJ, Andrade-Villanueva J, Girard PM, et al; ReAL Study Team. A randomized 
comparative 96-week trial of boosted atazanavir versus continued boosted protease 
inhibitor in HIV-1 patients with abdominal adiposity. Antivir Ther. 2012; 17(4): 689-700. 
6. 
Menard A, et al. Dolutegravir and weight gain: an unexpected bothering side effect? AIDS 
2017; 31:1499–1502. 
7. 
Norwood J, Turner M, Bofill C, Rebeiro P, Shepherd B, Bebawy S, Hulgan T, Raffanti S, Haas 
DW, Sterling TR, Koethe JR. Weight Gain in Persons With HIV Switched From Efavirenz-
Based to Integrase Strand Transfer Inhibitor-Based Regimens. J Acquir Immune Defic 
Syndr. 2017; 76(5):527-31.  
8.       
Gupta SK , Mi D, Liu Z, Saha C. Endothelial, inflammatory, coagulation, metabolic 
effectsand safety of etravirine in HIV-uninfected volunteers. AIDS Patient Care 
STDS. 2011; 25(6): 327-31. 

36 
CRF003 protocol version 4.0 20 Jan 2023 
9.          Pao VY, Lee GA, Taylor S, Aweeka FT, Schwarz JM, Mulligan K, Schambelan M, Grunfeld C. 
The 
protease 
inhibitor 
combination lopinavir/ritonavir 
does 
not 
decrease insulin secretion in healthy, HIV-seronegative volunteers. AIDS. 2010; 
24(2):265-70.10.        
10.      Dubé MP, Shen C, Greenwald M, Mather KJ. No impairment of endothelial function or 
insulin sensitivity with 4 weeks of the HIV protease inhibitors atazanavir or lopinavir-
ritonavir in healthy subjects without HIV infection: a placebo-controlled trial. Clin Infect 
Dis. 2008; 47(4): 567-74.11. 
 
11.        Elliot ER, Cerrone M, Challenger E, Else L, Amara A, Bisdomini E, Khoo S, Owen A, Boffito     
M. 
Pharmacokinetics 
of dolutegravir with 
and 
without 
darunavir/cobicistat 
in healthy volunteers.J Antimicrob Chemother. 2019; 74(5): 1466. 
12.     
McAllister  JW, Towns JM, Mculty A, Pierce AB, Foster R,  Richardson R, Carr A. 
Dolutegravir with tenofovir disoproxil fumarate-emtricitabine as HIV postexposure 
prophylaxis in gay and bisexual men. AIDS. 2017; 31(9): 1291-5. 
13. 
DeFronzo RA, Tobin JD, Andres R. Glucose clamp technique: a method for quantifying 
insulin secretion and resistance. Am J Physiol 1979; 237: E214-E23. 
14.  
Randell P, Jackson A, Milinkovic A, Boffito M, Moyle G. An open-label, randomized study   
of the impact on insulin sensitivity, lipid profile and vascular inflammation by treatment 
with lopinavir/ritonavir or raltegravir in HIV-negative male volunteers. Antivir Ther 2017; 
22: 145-51. 
15.          Noor MA, Parker RA, O'Mara E, Grasela DM, Currie A, Hodder SL, Fiedorek FT, Haas DW. 
The effects of HIV protease inhibitors atazanavir and lopinavir/ritonavir on insulin 
sensitivity in HIV-seronegative healthy adults. AIDS. 2004; 18(16): 2137-44. 
 
16.   
 Spinner CD , Schulz S,  Bauer U, Schneider J, Bobardt  J ,  Von Werder A,  Schmid 
A, ZinkA , Wolf E , Iakoubov R.  Effects of antiretroviral combination therapies F/TAF, 
E/C/F/TAF and R/F/TAF on insulin resistance in healthy volunteers: the TAF-IR Study. 
Antivir Ther. 2018; 23(7):629-32. 
 

37 
CRF003 protocol version 4.0 20 Jan 2023 
Appendices  
Appendix 1: Study flowchart  
 
 
Screen 
(-28) 
Baseline 
Day 
28 
Day 44 
Day 72 
Follow-up 
call (Day 
100) 
Informed 
consent 
√ 
 
 
 
 
 
Physical Exam 
√ 
 
 
 
 
 
Medical history 
√ 
 
 
 
 
 
Demographics 
√ 
 
 
 
 
 
Vital signs 
√ 
√ 
√ 
√ 
√ 
 
Weight 
√ 
√ 
√ 
√ 
√ 
 
Height, 
waist/hip 
measurement 
√ 
√ 
√ 
√ 
√ 
 
Adverse Events 
 
√ 
√ 
√ 
√ 
√  
Concomitant 
medication  
√ 
√ 
√ 
√ 
√ 
 
Urinary analysis1 
and Urine drug 
testing  
√ 
√ 
√ 
√ 
√ 
 
Biochemistry 2 
√3 
√ 
√ 
√ 
√ 
 
Haematology4 
√ 
√ 
√ 
√ 
√ 
 
Pregnancy 
testing 5 
√ 
√ 
√ 
√ 
√ 
√ 
Insulin 
 
√ 
√ 
 
√ 
 
TSH, fT4 
 
√ 
√ 
√ 
√ 
 
HIV, HBV &HCV  
√ 
 
 
 
 
 

38 
CRF003 protocol version 4.0 20 Jan 2023 
CRP 
 
√ 
√ 
√ 
√ 
 
Adoponectin, 
leptin, ghrelin 
 
√ 
√ 
√ 
√ 
 
Pituitary 
hormone tests  
 
√ 
√ 
√ 
√ 
 
Testosterone  
 
√ 
√ 
√ 
√ 
 
Euglycaemic 
clamp 
 
√ 
√ 
 
√ 
 
ECG 
√ 
√ 
√ 
 
√ 
 
Indirect 
calorimetry  
 
√6 
√7 
 
√8 
 
Dietary 
Questionnaire 
 
√ 
√ 
√ 
√ 
 
Sleep 
Questionnaires  
 
√ 
√ 
√ 
√ 
 
Self-
administered 
dosing 
 
√7 (am) 
√7 
√8 (am) 
√8 
 
Drug Dispensing   
√7 
 
√8 
 
 
Drug returns 
 
 
√7 
 
√8 
 
1-urinary analysis (pH, specific gravity, leucocytes, nitrite, protein, glucose, ketones, urobilinogen, bilirubin, 
erythrocytes)   
2-Biochemistry safety labs assessment for standard clinical chemistry (glucose, creatinine, sodium, 
potassium, calcium, aspartate aminotransferase, alanine aminotransferase, bilirubin), lipids (triglycerides, 
cholesterol, cholesterol HDL & LDL Sub-fractions) and Hb1Ac  
3-screening visit is not fasting visit  
4-haematology (haemoglobin, haematocrit, erythrocyte count, white blood count, differential white blood 
cell count, platelets)  
5- Pregnancy testing and LMP date in female subjects 
6- All participants 
7-only Group 1 
8-only Group 2 
 
 
 
 
 

39 
CRF003 protocol version 4.0 20 Jan 2023 
Appendix 2: Collecting, processing, shipping & storing of blood & plasma  
 
Processing blood for the measurement of adipocytokines  
For Leptin, Adiponectin and Ghrelin  
Please see the laboratory manual for full details.  
Appendix 3: Division of AIDS (DAIDS) Table for Grading the Severity of Adult and 
Pediatric Adverse Events 
 
Please reference latest version of grading scale available at below location. 
https://rsc.niaid.nih.gov/sites/default/files/daidsgradingcorrectedv21.pdf  
 
 
Appendix 4:  Risk Assessment:  Dolutegravir (TIVICAY) 
 
Potential Risk of 
Clinical 
Significance 
Data/Rationale for Risk 
Mitigation Strategy 
 
Investigational Product: dolutegravir 
Refer to the most recently approved SmPC for this trial  
Hypersensitivity 
and rash 
 
Hypersensitivity reactions have been 
reported with dolutegravir, and were 
characterized by rash, constitutional 
findings, and sometimes, organ 
dysfunction, including severe liver 
reactions. Further information on this 
risk is provided in Section 7.5. 
Subjects with history or 
presence of allergy/sensitivity to 
any of the study drugs or their 
components are excluded  
Specific/detailed toxicity 
management guidance is 
provided for hypersensitivity 
reactions and rash (Section 7.5) 
The subject informed consent 
form includes information on 
this risk and the actions subjects 
should take in the event of a 
hypersensitivity reaction or 
associated signs and symptoms. 
 Subjects will be monitored for 
symptoms of hypersensitivity 
reactions and stop treatment 
immediately should they appear. 
These symptoms include, 
but not limited to, severe rash or 
rash accompanied by raised liver 
enzymes, fever, general malaise, 
fatigue, muscle 

40 
CRF003 protocol version 4.0 20 Jan 2023 
Potential Risk of 
Clinical 
Significance 
Data/Rationale for Risk 
Mitigation Strategy 
 
Investigational Product: dolutegravir 
Refer to the most recently approved SmPC for this trial  
or joint aches, blisters, oral 
lesions, conjunctivitis, facial 
oedema, eosinophilia, 
angioedema). 
Clinical status including 
liver aminotransferases and 
bilirubin should also be 
monitored. Delay in stopping 
treatment with dolutegravir or 
other suspect active substances 
after the onset of 
hypersensitivity may result in a 
life-threatening allergic reaction. 
 
Drug induced 
liver injury and 
other clinically 
significant liver 
chemistry 
elevations 
Non-clinical data suggested a 
possible, albeit low, risk for 
hepatobiliary toxicity with DTG. Drug-
related hepatitis is considered an 
uncommon risk for antiretroviral 
therapy containing DTG regardless of 
dose or treatment population.  
 
 
 
 
Subjects meeting any of the 
following criteria during the 
screening period are excluded 
from participating  
 Alanine aminotransferase 
(ALT) or Aspartate 
aminotransferase (AST)  ≥1.5 
times the upper limit of 
normal (ULN) or total 
bilirubin ≥1.5xULN  
Positive for HBV (hepatitis B 
virus surface antigen positive 
[+HBsAg]) or positive HCV 
(positive hepatitis C antibody 
test) within 3 months of the Day 
1 study visit  
Specific/detailed liver stopping 
criteria and toxicity 
management guidance are 
provided for suspected drug 
induced liver injury or other 
clinically significant liver 
chemistry elevations  
Neural Tube 
Defects 
In the ongoing observational study 
conducted in Botswana, preliminary 
results show that in babies born to 
women who were taking DTG when 
Pregnancy testing should be 
performed before initiation 
of DTG therapy in all women of 
child bearing potential (negative 

41 
CRF003 protocol version 4.0 20 Jan 2023 
Potential Risk of 
Clinical 
Significance 
Data/Rationale for Risk 
Mitigation Strategy 
 
Investigational Product: dolutegravir 
Refer to the most recently approved SmPC for this trial  
they became pregnant there was an 
increased risk of neural tube defects 
compared with the background rate.  
Dolutegravir was tested in a complete 
package of reproductive toxicology 
studies, including embryofoetal 
development studies. No adverse 
development outcomes, including 
neural tube defects, were identified 
from reproductive toxicology studies. 
In reproductive toxicity studies in 
animals, DTG was shown to cross the 
placenta. 
Data available from other sources 
including the Antiretroviral Pregnancy 
Registry (APR), other cohorts and 
clinical trials are insufficient to 
confirm or refute this potential risk.   
pregnancy test at screening and 
randomization).  
 
Women who are pregnant or 
who plan to become pregnant 
are excluded.  
 
All women of reproductive 
potential should use effective 
contraception (Appendix 6). 
 
The subject informed consent 
form provides information about 
this potential risk. 
 
 
Risk Assessment: Clamp procedure 
 
The euglycaemic clamp procedure is associated with a potential risk of hypokalemia. In order to 
avoid this risk, whole blood potassium will be measured along with  electrocardiograph 
monitoring at 30 min intervals as a safety precaution to detect any tendency toward hypokalemia. 
 
 
Appendix 5: Liver Chemistry Stopping and Follow up Criteria 
 
Dolutegravir must be discontinued if: 
• 
AST or ALT≥3xULN.  
• 
 If AST or ALT≥3xULN AND bilirubin >2xULN (>35% direct bilirubin), i.e. Hy’s case, report 
event as SAE. 
 
Required Actions and Follow up Assessments following ANY Liver Stopping Event 
Actions 
Follow Up Assessments 

42 
CRF003 protocol version 4.0 20 Jan 2023 
 
Immediately discontinue DTG  
 
Subject should not be rechallenged due 
to the risk of a recurrent reaction.   
 
Report the event to Chelsea and 
Westminster NHS Trust R&D by telephone 
within 24 hours. 
 
Events of possible drug-induced liver injury 
with hyperbilirubinemia2 will be reported 
to Chelsea and Westminster NHS Trust 
R&D as serious adverse events using the 
serious adverse event case report form. 
 
Complete the liver event case report form 
for all events meeting liver stopping 
criteria, and submit to Chelsea and 
Westminster NHS Trust R&D within one 
week of first becoming aware of the event 
 
Perform liver event follow up assessments.  
 
Monitor the subject until liver chemistries 
resolve, stabilise, or return to within 
baseline  
 
Subject may continue in the study for any 
protocol specified follow up assessments. 
 
 
MONITORING: 
 
Make every reasonable attempt to have 
subjects return to clinic within 24 hours 
for repeat liver chemistries (include ALT, 
AST, alkaline phosphatase, bilirubin) and 
perform liver event follow up assessments 
at the central laboratory as described to 
the right.  
 
A specialist or hepatology consultation is 
recommended. 
 
Monitor subjects twice weekly until liver 
chemistries resolve, stabilise or return to 
within baseline. 
 
Viral hepatitis serology, including: 
 
Hepatitis A immunoglobulin M 
(IgM) antibody; 
 
HBsAg and hepatitis B core 
antibody; 
 
Hepatitis C RNA; 
 
Hepatitis E IgM antibody. 
 
Cytomegalovirus IgM antibody. 
 
Epstein-Barr viral capsid antigen IgM 
antibody (or if unavailable, obtain 
heterophile antibody or monospot 
testing). 
 
Syphilis screening. 
 
Drugs of abuse screen, including alcohol. 
Record alcohol use on the liver event 
case report form 
 
Serum acetaminophen adduct High 
Performance Liquid Chromatography 
assay (quantifies potential 
acetaminophen contribution to liver 
injury in subjects with definite or likely 
acetaminophen use in the preceding 
week [FDA, 2009]).  The site must 
contact the medical monitor when this 
test is required.  
 
Serum creatinine kinase and lactate 
dehydrogenase. 
 
Fractionate bilirubin, if total bilirubin 
1.5xULN. 
 
Obtain complete blood count with 
differential to assess eosinophilia. 
 
Anti-nuclear antibody, anti-smooth 
muscle antibody, Type 1 anti-liver 
kidney microsomal antibodies, and 
quantitative total immunoglobulin G (or 
gamma globulins). 
 
Liver imaging (ultrasound, magnetic 
resonance, or computerised 
tomography) and /or liver biopsy to 
evaluate liver disease. Complete Liver 

43 
CRF003 protocol version 4.0 20 Jan 2023 
Imaging and/or Liver Biopsy case report 
form. 
 
Record the appearance or worsening of 
clinical symptoms of liver injury, or 
hypersensitivity, fatigue, decreased 
appetite, nausea, vomiting, abdominal 
pain, jaundice, fever, or rash as relevant 
on the AE report form 
 
Record use of concomitant medications 
on the concomitant medications report 
form including acetaminophen, herbal 
remedies, other over the counter 
medications, and alcohol use. 
Note: 
Serum bilirubin fractionation should be performed if testing is available. If serum bilirubin 
fractionation is not immediately available, discontinue study treatment for that subject if ALT or 
AST 3xULN and bilirubin  2xULN. Additionally, if serum bilirubin fractionation testing is 
unavailable, record presence of detectable urinary bilirubin on dipstick, indicating direct bilirubin 
elevations and suggesting liver injury.  
All events of ALT or AST  3xULN and bilirubin 2xULN (>35% direct bilirubin) or ALT or AST3xULN 
and INR>1.5, if INR measured which may indicate severe liver injury must be reported as a serious 
adverse event 
 
Appendix 6:Highly Effective Methods for Avoiding Pregnancy in Women of Child 
Bearing Potential* 
 
The following is the all-inclusive list of the highly effective methods for avoiding pregnancy (i.e., 
have a failure rate of less than 1% per year when used consistently and  correctly and, when 
applicable, in accordance with the product label). 
The list does not apply to FCBP with same sex partners, when this is their preferred and usual 
lifestyle. Periodic abstinence (e.g. calendar, ovulation, symptothermal, post-ovulation methods) 
and withdrawal are not acceptable methods of contraception. 
• 
Contraceptive subdermal implant 
• 
Intrauterine device or intrauterine system 
• 
Combined estrogen and progestogen oral contraceptive [Hatcher, 2011]) 
• 
Injectable progestogen [Hatcher, 2011] 
• 
Contraceptive vaginal ring [Hatcher, 2011] 
• 
Percutaneous contraceptive patches [Hatcher, 2011] 
• 
Male partner sterilization with documentation of azoospermia prior to the female 
subject's entry into the study, and this male is the sole partner for that subject [Hatcher, 2011]. 
The information on the male sterility can come from the site personnel’s: review of subject’s 
medical records; medical examination of the subject and/or semen analysis; or medical history 
interview provided by her or her partner. 

44 
CRF003 protocol version 4.0 20 Jan 2023 
•  
Male participants who have partners who are women of childbearing potential must use 
a condom throughout the study and for a period of at least 4 weeks after the study;  in addition 
to any other methods used to avoid partner pregnancy. 
 
* A Women of childbearing potential (WOCBP) is defined as any female who has experienced 
menarche and who has not undergone surgical sterilization (hysterectomy or bilateral 
oophorectomy) and is not postmenopausal, females under the age of 55 years must have a 
serum follicle stimulating hormone, (FSH**) level > 40mIU/mL to confirm menopause. 
** Females treated with hormone replacement therapy, (HRT) are likely to have artificially 
suppressed FSH levels and may require a washout period in order to obtain a physiologic FSH 
level.  
 
Ref: Hatcher RA, Trussell J,  Nelson AL, Cates W Jr, Stewart F, Kowal D, Policar MS, editors. 
Contraceptive Technology. 20th edition.  Atlanta, Georgia: Ardent Media, Inc., 2011: 50.  Table 3 
2.