Single dose fed 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 TitleSingle dose Fed 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-1005
ProductEmtricitabine, Tenofovir Alafenamide and Dolutegravir 15 mg/1.88 mg/5 mg Tablets for Oral Suspension
Study TypePivotal Fed Bioequivalence
Version00
Protocol Date10 Sep 2025
General DescriptionThis 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 fed pharmacokinetics will be characterized in Fourty-eight (48) 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, seven milliliter (1 × 7 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, three milliliter (1 × 3 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. All aliquots (Four aliquots of period-1 & 2 and two aliquots of period-3 & 4) of plasma will be extracted and stored in suitably labeled 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 412 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 for the pharmacokinetic parameters Cmax, AUC0-t 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 fed 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.

Note: The intended commercial product name for emtricitabine/tenofovir 15/1 .88 mg tablets for oral suspension is Descovy; however, that name is not approved for this product. For the purposes of these clinical studies, the labels will contain "emtricitabine/tenofovir 15/1.88 mg tablets for oral suspension instead of DESCOVY®.

Manufactured by: Cambrex (Mirabel, Quebec, Canada), 17800 Lapointe, Mirabel, Quebec,J7J 0W8-Canada

Manufactured for: 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 fed pharmacokinetics will be characterized in Fourty-eight (48) 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 fed 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.  |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 enrollment.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, urine pregnancy test (for female subjects only) 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. |HousingEnough 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 11.00 hours prior until at least 72.00 hours after dosing for period-1 & period-2. Subjects will be housed 11.00 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 ManagementSerum 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) Creatinine clearance (CrCl) test will be done by using Cockcroft-Gault method will be performed during screening and post study. |Bioanalytical MethodA 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 DeterminationAll 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 8.5.2 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.

Primary Pharmacokinetic Parameters:

Cmax: Maximum observed plasma concentration

AUC0 t: The area under the plasma concentration versus time curve from time zero to the last measurable concentration

Secondary Pharmacokinetic Parameters:

Tmax: Time of the maximum measured plasma concentration

AUC0 inf: The area under the plasma concentration versus time curve from time zero to infinity. Where AUC0-inf = AUC0 t  Ct/ Kel, Ct is the last measurable concentration and z is the terminal elimination rate constant

t½: The elimination half-life will be calculated as 0.693/ Kel

Kel: First order elimination rate constant associated with the terminal (loglinear) portion of the curve. This is estimated via linear regression of time vs. log concentration. This parameter will be calculated by linear least squares regression analysis using at least last three or more non-zero plasma concentration values.

For Tenofovir: [Considering the half life > 24hrs]

Primary Pharmacokinetic Parameters:

Cmax: Maximum observed plasma concentration

AUC0-72: The area under the plasma concentration versus time curve from time zero to the last measurable concentration

Secondary Pharmacokinetic Parameters:

Tmax: Time of the maximum measured plasma concentration

|Statistical Analysis of the Pharmacokinetic Parameters Pharmacokinetic parameters data obtained for the analytes Emtricitabine, Tenofovir Alafenamide, Tenofovir and Dolutegravir will be included in the statistical analysis as per the criteria mentioned below by using SAS version 9.4 or higher. For Emtricitabine, Dolutegravir, Tenofovir: Pharmacokientic parammeters data of the subjects who completes the first two periods of the study will be considered for statistical analysis.

For Tenofovir Alafenamide:

• 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 complete all periods of the study will be considered in average bioequivalence evaluation as per IEC approved protocol using SAS version 9.4 or higher.

Descriptive statistics of all the pharmacokinetic parameters will be computed 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.

The 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 ln-transformed pharmacokinetic parameters Cmax and AUC0-t.of Emtricitabine, Tenofovir (Cmax, AUC0-72) and Dolutegravir will be subjected to Analysis of Variance (ANOVA) by using PROC GLM procedure of SAS version 9.4 or higher. 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 main effects will be tested at 5% Level of Significance.

Bioequivalence evaluation:

BE criteria For Emtricitabine, Tenofovir and Dolutegravir: 90% CI should fall between 80 to125% for all primary pharmacokinetics for Emtricitabine & Dolutegravir.

Note: Pharmacokinetic data from tenofovir will be provided as supportive evidence of comparable therapeutic outcome.

Bioequivalence evaluation:

BE criteria for Tenofovir Alafenamide:

1. If within-subject standard deviation of the reference product SWR  0.294 then assessment of bioequivalence will be determined using scaled average bioequivalence criteria.

a. The point estimates (Geometric Least Squares mean of test/reference ratios) must fall within the bioequivalent limits 80.00% 125.00%.

b. The 95% upper confidence bound for (T- R)2- S2 WR must be 0.0000, where  = (ln (1.25)/ W0)2 andW0 = 0.25.

2. If within-subject standard deviation of the reference product SWR < 0.294 then assessment of bioequivalence will be determined using average bioequivalence criteria

|**APPENDIX VI:**STUDY CONDUCT INFORMATION Dr. Nikhil Kumar Kursam., M.B.B.S., M.D.,CLINICAL DEVELOPMENT DIVISION,

Aizant Drug Research Solutions private limited.,

Survey No.: 172 &173, Apparel Park Road, Dulapally Village,

Dundigal Gandimaisamma Mandal,

Medchal-Malkhajgiri District - 500100,

Phone No.: +91 40 23792190/91/92;

Fax No.: +91 40 23792223.

Clinical Research Facility:

Clinical Pharmacology Unit-I,

CLINICAL DEVELOPMENT DIVISION

Aizant Drug Research Solutions private limited.,

Survey No.: 172 &173, Apparel Park Road, Dulapally Village,

Dundigal Gandimaisamma Mandal,

Medchal-Malkhajgiri District - 500100,

Phone No.: +91 40 2379219

Stastical Investigator and Randomization Facility:

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. Phone No.: +91 40

23792190/91/92, Fax No.: +91 40 23792223

e-mail ID: udaya.konda@aizant.com

Institutional Review Board:

Maarg Independent Ethics Committee

Address: H. No.: 8-3-721/11/6/1/2, Maram Reddy Nivas, 2nd Floor, Beside

PJR statue,Srinagar Colony, Hyderabad, India-500073.

Ph No: 8885574599.

Clinical Laboratory Facility-1:

Aizant Drug Research Solutions Pvt. Ltd., Diagnostics Department

Clinical Development Division,

Aizant Drug Research Solutions Pvt. Ltd.,

Survey No.: 172 &173, Apparel Park Road, Dulapally Village,

Dundigal Gandimaisamma Mandal, Medchal-Malkhajgiri District-500100

Telangana, India

+91 40 23792190/91/92

Clinical Laboratory Facility-2:

Lucid Medical Diagnostics Pvt. Ltd.,

Plot No: 1299-E, Road No. 68, Jubilee Hills,

Co Op House Building Ltd.

Hyderabad -500033

+91 9603892224

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