Protocol Title

Single dose Fasting oral bioequivalence study of Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension (T) of Mylan Laboratories Limited, India with that of Reference product (R=R1+R2) R1: TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension of ViiV healthcare, Durham, NC 27701 and R2: DESCOVY® (emtricitabine and tenofovir alafenamide) 15 mg and 1.88 mg Tablets for oral suspension of Gilead Sciences, Inc. Foster City, CA 94404 in healthy adult males and non-pregnant, non-lactating females.

Protocol No.

TAED-TBP-1002

Product

Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg

Tablets for Oral Suspension

Study Type

Pilot Fasting Bioequivalence

Version

03                     Region of Submission:  PEPFAR

Protocol Date

07 May 2025

General Description

This protocol describes an open labelled, balanced, single-dose, randomized, two-treatment, four-period, two-sequence, crossover, full replicate study to investigate the bioequivalence of Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension (T) of Mylan Laboratories Limited, India with that of Reference product (R=R1+R2) R1: TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension of ViiV healthcare, Durham, NC27701 and R2: DESCOVY® (emtricitabine and

tenofovir alafenamide) 15 mg and 1.88 mg Tablets for oral suspension of Gilead Sciences, Inc. Foster City, CA 94404.

Single dose fasting pharmacokinetics will be characterized in Twenty-four (24) healthy adult males and non-pregnant, non-lactating females following administration of a single oral dose of either test product (T) or reference product (R=R1+R2) as per randomization schedule. The detailed administration process is given in the section 7.5. Treatment procedure of the protocol.

For Period 01 & 02: For Emtricitabine Tenofovir Alafenamide, Tenofovir and Dolutegravir:

In each study period, Six milliliter (1 × 6 mL) blood samples (23) will be collected in K3EDTA Vacutainers at pre-dose (0.00 hour) and the following times after dosing: 0.083, 0.17, 0.33, 0.50, 0.75, 1.00, 1.25, 1.50, 1.75, 2.00, 2.50, 3.00, 3.50, 4.00, 6.00, 8.00, 10.00, 12.00, 24.00, 36.00, 48.00 and 72.00 hours.

 For Period 03 & 04: For Tenofovir Alafenamide:

In each study period, four milliliter (1 × 4 mL) blood samples (15) will be collected in K3EDTA Vacutainers at pre-dose (0.00 hour) and the following times after dosing: 0.083, 0.17, 0.33, 0.50, 0.75, 1.00, 1.25, 1.50, 1.75, 2.00, 2.50, 3.00, 3.50, 4.00 hours post dose.

 The Collected blood samples will be placed in an ice batch and centrifuged under refrigeration as soon as possible. Two aliquots (Aliquot 1 of 2 & Aliquot 2 of 2) of plasma will be extracted and stored in suitably labelled tubes at -70°C or colder with an acceptable operating range within -55°C to -90°C at the clinical site until transferred on dry ice to analytical site (refer Appendix-I: Bioanalytical sample handling instruction of the protocol).

For the determination of the pharmacokinetic disposition of the formulations, there will be a total of 76 (46 blood samples from period-1 & 2 and 30 blood samples from period-3 & 4) blood samples involving a total of 396 mL of blood collected for pharmacokinetic analysis from each subject in the study. There will be at least 07 days gap between dosing times for the treatment periods.

The bioequivalence of Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension (T) of Mylan Laboratories Limited, India with that of Reference product (R=R1+R2) R1: TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension of ViiV healthcare, Durham, NC27701 and R2: DESCOVY® (emtricitabine and tenofovir alafenamide) 15 mg and 1.88 mg Tablets for oral suspension of: Gilead Sciences, Inc. Foster City, CA 94404 will be assessed by a statistical comparison of various pharmacokinetic parameters derived from the plasma concentration-time curves of the drug.

OBJECTIVES

Primary Objective: To investigate the bioequivalence of Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension (T) of Mylan Laboratories Limited, India with that of Reference product (R=R1+R2) R1: TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension of ViiV healthcare, Durham, NC27701 and R2: DESCOVY® (emtricitabine and tenofovir alafenamide) 15 mg and 1.88 mg Tablets for oral suspension of Gilead Sciences, Inc. Foster City, CA 94404. in healthy adult males and non-pregnant, nonlactating females.

Secondary Objective: To evaluate the safety of the subjects (i.e. monitor adverse events) under fasting conditions.

STUDY DRUG

Test product (T): Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension,

Manufactured by: Mylan Laboratories Limited, India.

Reference product (R=R1+R2):

Reference product (R1): TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension

Manufactured for: ViiV healthcare, Durham, NC 27701.

Reference Product (R2): DESCOVY® (emtricitabine and tenofovir alafenamide) 15 mg and 1.88 mg Tablets for oral suspension

Manufactured by: Manufactured by: Cambrex (Mirabel, Quebac, Canada), 17800 Lapointe, Mirabel, Quebec, J7J0W8- Canada.

Distributed by: Gilead Sciences, Inc. Foster City, CA 94404.

STUDY CONDUCT

This is an open labelled, balanced, single-dose, randomized, two-treatment, four-period, two sequence, crossover, full replicate study to investigate the bioequivalence of Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension (T) of Mylan Laboratories Limited, India with that of Reference product (R=R1+R2) R1: TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension of ViiV healthcare, Durham, NC27701 and R2: DESCOVY® (emtricitabine and tenofovir alafenamide) 15 g and 1.88 mg Tablets for oral suspension of Gilead Sciences, Inc. Foster City, CA 94404 in healthy adult males and nonpregnant, non-lactating females.

Single dose fasting pharmacokinetics will be characterized in Twenty-four (24) healthy adult males and non-pregnant, non-lactating females following  administration of a single oral dose of either test product (T) or reference product (R=R1+R2) under fasting conditions.

Subjects will be housed the evening prior to drug dosing of period-1, period-2 and until at least 72.00 hours after investigational drug administration. Blood samples will be collected prior to dosing and for up to 72.00 hours after period-1 and period-2 dosing. Subjects will be housed the evening prior to drug dosing of period-3, period-4 and until at least 24.00 hours after investigational drug administration. Blood samples will be collected prior to dosing and for up to 4.00 hours after period-3 and period-4 dosing. There will be at least 07 days gap between dosing times of each treatment periods. Individual subjects should be dosed at approximately the same time of day in each dosing period. It is the sponsor’s intent to complete all subjects simultaneously. Thus, it is anticipated that all subjects will be completed in a single cohort within approximately 24 days following the initiation of dosing. If multiple cohorts are necessary to enroll the required number of subjects, no two cohorts can be dosed on the same day.

Screening Procedures

Each prospective subject must agree to participate in screening procedures by signing the most recent Institutional Review Board/Independent Ethics Committee approved Informed Consent Document (ICD) before any screening procedure is initiated. The Principal Investigator or Medical Sub-Investigator will review the inclusion and exclusion criteria to confirm eligibility of each subject prior to enrolment.

Each subject will undergo a screening procedure for health assessment, which consists of a complete medical history, physical examination with vital signs, clinical laboratory evaluations, 12-lead ECG and Chest X-ray PA view. The physical examination findings, ECG, and the laboratory tests can be considered as valid for maximum of 21 days prior to the dosing (drug administration) in first period of the study. Chest X-ray PA view will be taken within 6 months prior to dosing (drug administration) of period-1.

The physician in charge will assess abnormal values to determine if it is clinically significant.

Clinical/Laboratory Diagnostic Tests:

1. Haematology: Red blood cell count, White blood cell count, Differential white blood cell count (Neutrophils%, Lymphocytes%, Eosinophils%, Monocytes% and Basophils%), Hemoglobin estimation, hematocrit (HCT), Platelet count, Blood grouping and Rh typing.

2.  Biochemistry: Serum creatinine, Blood urea, SGOT (AST), SGPT (ALT), Serum alkaline phosphatase (ALP), Total bilirubin, Blood sugar / Plasma Glucose (Random) and Serum electrolytes (Sodium, Potassium and Chloride).

3. Serology: HIV (1 & 2) antibodies, Hbs Ag (Hepatitis B surface antigen) and HCV antibodies and VDRL/Syphilis

4. Urine analysis: Color, Appearance/Transparency, pH, Specific gravity, Glucose, Ketones, Bilirubin, Blood, Leucocytes, Proteins, Nitrite, Urobilinogen and Urine microscopic examination (Pus Cells, Red Blood Cells, Epithelial Cells, Casts and Crystals).

5.  Additional tests: Serum creatinine clearance (CrCl) test will be done at the time of screening by using Cockcroft-Gault method.

Housing

Enough subjects from the general population will be available in the clinic for Period-1 dosing in order to dose the required number of subjects. Subjects will be housed 10.50 hours prior until at least 72.00 hours after dosing for period-1 & period-2. Subjects will be housed 10.50 hours prior to dosing until at least 24.00 hours after dosing for period-3 and period-4. There will be at least 07 days gap between dosing times for the treatment periods.

Toxicity Management

Bioanalytical Method

A validated assay method will be employed for the analysis of Emtricitabine, Tenofovir Alafenamide, tenofovir and Dolutegravir in plasma samples. Full validation of the method, including precision, accuracy and reproducibility will be included in the final report, along with a statement regarding the stability of frozen samples.

Pharmacokinetic Parameter Determination

All concentration values below the lower limit of quantification (BLOQ) will be set to zero. Any missing samples will be reported as ‘M’ and will not be included for pharmacokinetic and statistical analysis.

The following pharmacokinetic parameters will be computed by using Phoenix® WinNonlin® version 8.0 or higher for Emtricitabine, Tenofovir Alafenamide, tenofovir and Dolutegravir through non compartmental method. Pharmacokinetic data from tenofovir will be provided as supportive evidence of comparable therapeutic outcome. Subjects concentration data from the first two periods will be considered for the pharmacokinetic parameters estimation of Emtricitabine, Tenofovir & Dolutegravir:

For TAF: The pharmacokinetic parameters will be estimated by using the concentration data up to 04.00 hrs post dose for all four periods.

Statistical Analysis of the Pharmacokinetic

Parameters

Subjects who completed all periods of the study will be included in scaled average BE and within reference variability estimation. All other scenarios where the subject didn’t completed all periods of the study will be considered in average bioequivalence evaluation as per IEC approved protocol using SAS version 9.4 or higher version. Descriptive statistics of all the pharmacokinetic parameters will be computed and reported for Emtricitabine, Tenofovir Alafenamide, Tenofovir and Dolutegravir.

The below will be calculated and reported for Emtricitabine, Tenofovir Alafenamide, Tenofovir and Dolutegravir:

The summary statistics (for relevant pharmacokinetic parameters) will be computed and reported for both test and reference products. Two one-sided test for bioequivalence and 90% confidence intervals for the ratio of least squares mean between drug formulations will be calculated, for ln-transformed data of Cmax and AUC0-t. Ratio of least squares means of test and reference products will be computed for ln-transformed pharmacokinetic parameters Cmax, and AUC0-t. Ratio summarizations will be reported for all non-transformed pharmacokinetic parameters. Intra-Subject variability will be computed for ln-transformed pharmacokinetic parameters Cmax, and AUC0-t.

Statistical analysis plan for Emtricitabine, Tenofovir and Dolutegravir: The pharmacokinetic parameters data of the first two periods will be considered for statistical analysis. The ln-transformed pharmacokinetic parameters Cmax and AUC0-t of Emtricitabine, Tenofovir and Dolutegravir will be subjected to Analysis of Variance (ANOVA). ANOVA model will include Sequence, Formulation, Period and Subject (Sequence) as fixed effects. Sequence effect will be tested using Subject (Sequence) as an error term. An F-test will be performed to determine the statistical significance of the effects involved in the model at a significance level of 5% (alpha =0.05). All the period, Treatment and sequence effects will be tested at 5% Level of Significance.

Bioequivalence evaluation: BE criteria For Emtricitabine, Tenofovir and Dolutegravir: 90% CI should fall between 80- 125% for all primary pharmacokinetic parameters for Emtricitabine & Dolutegravir. Statistical analysis plan for Tenofovir Alafenamide: The pharmacokinetic parameters data of the subjects who have PK data available for all four periods will be used in the estimation of within reference standard deviation. Subjects who received at least One Test and One Reference (R=R1+R2) will be included in the bioequivalence evaluation if average BE is conducted. The ln-transformed pharmacokinetic parameters Cmax and AUC0-t of Emtricitabine, Tenofovir and Dolutegravir will be subjected to Analysis of Variance (ANOVA). ANOVA model will include Sequence, Formulation, Period and Subject (Sequence) as fixed effects. An F-test will be performed to determine the statistical significance of the effects involved in the model at a significance level of 5% (alpha =0.05). All the period, Treatment and sequence effects will be tested at 5% Level of Significance.

APPENDIX VI: STUDY CONDUCT

INFORMATION

The clinical research organization conducting this study on behalf of Mylan Laboratories Ltd., India is required to complete this sheet and forward to Mylan prior to the enrollment of any subjects into the study. Any updates throughout the study conduct period must be reported on this sheet.

Protocol Number: TAED-TBP-1002

Protocol Title: Single dose Fasting oral bioequivalence study of Emtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral

Suspension (T) of Mylan Laboratories Limited, India with that of Reference product (R=R1+R2) R1: TIVICAY PD (Dolutegravir) 5 mg Tablets for oral suspension of ViiV healthcare, Durham, NC27701 and R2: DESCOVY® (emtricitabine and tenofovir alafenamide) 15 mg and 1.88 mg Tablets for oral suspension of Gilead Sciences, Inc. Foster City, CA 94404 in healthy adult males and non-pregnant, non-lactating females.

Principal Investigator: Dr. Anjum Jabeen, M.B.B.S., M.D.,

Address of Principal Investigator: Aizant Drug Research Solutions Pvt. Ltd.,

Clinical Pharmacology Unit-II, Maitrivihar Commercial Complex, Block No.: 101-107 & 201-206, Ameerpet, Hyderabad, Telangana, India - 500016.

Clinical Research Facility: Aizant Drug Research Solutions Pvt. Ltd.,

Clinical Pharmacology Unit-II, Maitrivihar Commercial Complex, Block No.: 101-107 & 201-206, Ameerpet, Hyderabad, Telangana, India - 500016.

Stastical Investigator: Mr. Udaya Kumar Konda, Pharmacokinetic & Statistical analysis, Aizant Drug Research Solutions Pvt. Ltd., Survey No.: 172 &173, Apparel Park Road, Dulapally Village, Dundigal Gandimaisamma Mandal, Medchal-

Malkhajgiri District - 500100, Telangana, India.

Randomization Facility: Pharmacokinetic & Statistical analysis, Aizant Drug Research Solutions Pvt. Ltd., Survey No.: 172 &173, Apparel Park Road, Dulapally Village, Dundigal Gandimaisamma Mandal, Medchal- Malkhajgiri District - 500100, Telangana, India.

Bioanalytical Facility: Mylan Laboratories Limited, Clinical Research

Centre, Saradhi Chambers, Plot No.: 4-A, Beside Poulomi Hospital, Rukminipuri, Dr. A. S. Rao Nagar, Hyderabad, India 500062.

Medical Monitor: Dr. Mohna Mukund Toro, MD , Manager,

Global Clinical Sciences, Mylan Pharmaceuticals Pvt. Ltd., (Viatris Inc.,)

Prestige Tech Park, Platina-3, 7th to 12th floor, Prestige Tech Park,

Kadubeesanahalli, Bangalore -560103. Or Dr. Ramesh K S, MD, Deputy General Manager, Global Clinical Sciences., Mylan Pharmaceuticals Pvt. Ltd., (Viatris Inc.,)

Prestige Tech Park, Platina-3, 7th to 12th floor, Prestige Tech Park, Kadubeesanahalli, Bangalore -560103