Single dose Fed oral bioequivalence study of Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg of Mylan Laboratories Limited, India with Reference product (R= R1 + R2) R1: Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne Pharma Greenville, NC 27834 in healthy adult female subjects.”

Protocol Title Single dose Fed oral bioequivalence study of Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg of Mylan Laboratories Limited, India with Reference product (R= R1 + R2) R1: Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne Pharma Greenville, NC 27834 in healthy adult female subjects.
Protocol No. TRLV-TBZ-1010
Product Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets, 200 mg/300 mg/0.15 mg/0.03 mg
Study Type Pivotal Fed Bioequivalence
Version 01 Region of Submission USFDA- WHO
Protocol Date 09 Janu 2024
General Description This protocol
describes an open labelled, single-dose, randomized, balanced, two-period,
two-sequence, two-treatment, two-way, crossover study to investigate the
bioequivalence of Test product (T) Emtricitabine, Tenofovir Disoproxil
Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15
mg/0.03 mg of Mylan Laboratories Limited, India with Reference product (R= R1 +
R2) R1: Truvada®
(emtricitabine
and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for:
Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel
and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne
Pharma Greenville, NC 27834. Single dose fed
pharmacokinetics will be characterized in Fifty-four (54) healthy adult female
subjects following administration of a single oral dose of either test product
(T) or reference product (R= R1 + R2) as per randomization schedule.

 In each study period, Six milliliter (1 × 6 mL) blood samples (23) will be collected in K2EDTA Vacutainers at pre-dose (0.00) and the following times after dosing 0.25, 0.50, 0.75, 1.00, 1.50, 2.00, 2.50, 3.00, 3.50, 4.00, 4.50, 5.00, 5.50, 6.00, 8.00, 10.00, 12.00, 16.00, 24.00, 36.00, 48.00 and 72.00 hours. The Collected blood samples will be placed in an ice bath and centrifuged under refrigeration as soon as possible. Three aliquots (aliquot 1 of 3, aliquot 2 of 3 and aliquot 3 of 3) 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. For the determination of the pharmacokinetic disposition of the formulations, there will be a total of 46 (23 in each period) blood samples involving a total of 276 mL of blood collected for pharmacokinetic analysis from each subject in the study. There will be at least 14 days gap between dosing times for the treatment periods. The bioequivalence of Test product (T) Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg of Mylan Laboratories Limited, India with Reference product (R= R1 + R2) R1: Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne Pharma Greenville, NC 27834 will be assessed by a statistical comparison of various pharmacokinetic parameters derived from the plasma concentrationtime curves of the drug.

|  OBJECTIVES The objective of this study is to investigate the bioequivalence of Test product (T) Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg of Mylan Laboratories Limited, India with Reference product (R= R1 +R2) R1: Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne Pharma Greenville, NC 27834 in healthy adult female subjects and monitor the adverse events and to ensure the safety of the subjects under fed conditions. | STUDY DRUGInvestigational Drug: Test product (T): Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg, Manufactured by: Mylan Laboratories Limited, India. Reference product (R= R1+R2): (R1): Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg, Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany. (R2): Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg, Distributed by: Mayne Pharma Greenville, NC 27834. Manufactured by: Patheon, Inc., Mississauga, Ontario L5N 7K9, CANADA. | STUDY CONDUCT This is an open labelled, single-dose, randomized, balanced, two-period, two-sequence, two-treatment, two-way, crossover study to investigate the bioequivalence of Test product (T) Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg of Mylan Laboratories limited, India with Reference product (R=R1 + R2) R1: Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne Pharma Greenville, NC 27834 in healthy adult female subjects. Single-dose fed pharmacokinetics will be characterized in Fifty-four (54) healthy adult female subjects following administration of a single oral dose of either test product or reference product will be administered under fed conditions. Subjects will be housed in the clinical facility for at least 10.50 hours prior to dosing of each period and until at least 72.00 hours after investigational drug administration. Blood samples will be collected prior to dosing and for 72.00 hours after each dosing. There will be at least 14 days between dosing times for the 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 18 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 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, serum pregnancy test 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: Hematology: Red blood cell count, White blood cell count, Differential white blood cell count (Neutrophils%, Lymphocytes%, Eosinophils%, Monocytes % and Basophils %), Hemoglobin estimation,  Platelet count, Blood grouping and Rh typing. Prothrombin time (PT), activated partial thromboplastin time (aPTT) and International normalized ratio (INR). 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) Serology: HIV (1 & 2) antibodies, Hbs Ag (Hepatitis B surface antigen) and HCV antibodies. 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).  Pregnancy test for females: Serum β-HCG pregnancy test Additional tests: Serum Lactate, creatinine clearance | 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 11.00 hours prior to dosing until at least 72.00 hours after dosing. | Bioanalytical Method A validated assay method will be employed for the analysis of Emtricitabine, tenofovir, levonorgestrel and ethinyl estradiolin 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. Repeat analysis and ISR will be performed as per respective inhouse SOPs of designated bioanalytical facility. Specific details of recovery, precision and accuracy, specificity, selectivity and sensitivity of the assay will be included in the final report. If clinical part conducts in group, the bioanalysis will be initiated after completion of each group, but plasma concentration data will be released to PK/Stats after completion of all subjects in study. | Pharmacokinetic Parameter Determination All concentration values below the lower limit of quantification (BLOQ) will be set to zero, concentration obtained as 0, <0. No Peak will be reported as Zero for all pharmacokinetic and statistical calculations. 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 version for Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol through non compartmental method.   Note: 1: For all the above computations, actual time points of the sample collection will be used in case of sample collection deviations. 2: For an acceptable Kel value, r should not be less than 0.8944 (or r2 not less than 0.80). If r is less than 0.8944, then Kel is set missing. 3: If the pre-dose value is greater than 5% of Cmax, in any period then that subject/period data will be dropped from pharmacokinetics and statistical analysis. 4: If any subject experiences emesis at or within two times the median Tmax (2 × 3.25 hours = 6.5 hours) following the drug administration in each study period, such subject data will be analyzed however data of the subject will not be included in pharmacokinetic and statistical analysis. | Statistical Analysis of the Pharmacokinetic Parameters Statistical analysis will be performed on the data obtained from the subjects who completed the study 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 Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol. The summary statistics (for relevant pharmacokinetic parameters) will be computed and reported for both test and reference products of Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol. The ln-transformed pharmacokinetic parameters Cmax, AUC0-t and AUC0-inf of Emtricitabine, tenofovir and ethinyl estradiol and Cmax & AUC0-72 of Levonorgestrel 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. 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, AUC0-t and AUC0-inf of Emtricitabine, tenofovir and ethinyl estradiol and Cmax & AUC0-72 of Levonorgestrel. The power of a test to detect 20% difference between test and reference products will be computed and reported for Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol. Ratio of least squares means of test and reference products will be computed for ln-transformed pharmacokinetic parameters Cmax, AUC0-t and AUC0-inf. Ratio analysis will be reported for ln-transformed pharmacokinetic parameters Cmax, AUC0-t and AUC0-inf of Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol. Intra-Subject variability will be computed for ln-transformed pharmacokinetic parameters Cmax, AUC0-t and AUC0-inf of Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol. BE criteria  Emtricitabine, tenofovir, levonorgestrel and ethinyl estradiol: The 90% geometric confidence intervals of the ratio (T/R or A/B) of least-squares means from the ANOVA of the natural log transformed Cmax, AUC0-t and AUC0-inf should be within 80.00% to 125.00% for emtricitabine, tenofovir and ethinyl estradiol. Levonorgestrel: The 90% geometric confidence intervals of the ratio (T/R or A/B) of least squares means from the ANOVA of the natural log transformed Cmax and AUC0-72 should be within 80.00% to 125.00% for levonorgestrel.  Determination of biologically implausible (pharmacokinetic) outliers will be based on the observed results (concentration data; such as no detectable plasma concentration following the administration of drug) which are determined to be discordant with the rest of the subjects. Clinical and bioanalytical circumstances will be examined in an attempt to provide insight into the apparent anomalous results. It is the sponsor’s intent to complete this study in one cohort. In the event that separate enrollments are necessary to complete the intended number of subjects, appropriate adjustments may be made to the statistical model to reflect the multi-cohort nature of the study. a. Additional cohorts will be recruited and dosed under the following conditions: i. Recruiting for additional enrollment(s) begin before the end of the final period of the previous enrollment; ii. Subjects are recruited from the same population, under the same protocol requirements; iii. dosing of additional cohorts began as soon as practical after their recruitment; and iv. no two cohorts are dosed on the same day.   b. Appropriate adjustments will be made to the statistical model to reflect the multi-cohort nature of the study if: i. the number of cohorts is less than or equal to 3; and ii. there are at least 6 subjects in each cohort |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: TRVL-TBZ-1010 Protocol Title: Single dose Fed oral bioequivalence study of Emtricitabine, Tenofovir Disoproxil Fumarate, Levonorgestrel and Ethinyl Estradiol Tablets 200 mg/300 mg/0.15 mg/0.03 mg of Mylan Laboratories Limited, India with Reference product (R= R1 + R2) R1: Truvada® (emtricitabine and tenofovir disoproxil fumarate) tablets 200 mg / 300 mg Manufactured for: Gilead Sciences Inc, Foster City, CA 94404, Made in Germany and R2: Levora® 0.15/30-28 (Levonorgestrel and Ethinylestradiol Tablets USP), 0.15 mg /0.03 mg Distributed by: Mayne Pharma Greenville, NC 27834 in healthy adult female subjects. |Principal Investigator:(please attach curriculum vitae when return to Mylan) Dr. Nikhil Kumar Kursam, M.B.B.S, M.D., Aizant Drug Research Solutions Pvt. Ltd., Survey No.: 172 &173, Apparel Park Road, Dulapally Village, Dundigal Gandimaisamma Mandal, MedchalMalkhajgiri District - 500100, Telangana, India. Phone No.: +91 40 23792190/91/92, Fax No.: +91 40 23792223 Address of Principal Investigator +91 40 23792190/91/92 Phone Number of Principal Investigator |Clinical Research Facility: Clinical Pharmacology Department, Name 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. Phone No.: +91 40 23792190/91/92, Fax No.: +91 40 23792223 |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. Phone No.: +91 40 23792190/91/92, Fax No.: +91 40 23792223 e-mail ID: udaya.konda@aizant.com |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. Phone No.: +91 40 23792190/91/92, Fax No.: +91 40 23792223 e-mail ID: udaya.konda@aizant.com |Clinical data management and CDISC Facility: Inductive Quotient Analytics India private Limited. 6 th Floor, Krishe Sapphire Metro Station Durgamma, Cheruvu, Sri Sai Nagar, Madhapur, Hyderabad, Telangana 500081 Phone 040-69067400; +91 91541 98743; 91 810625012 |Institutional Review Board: Maarg Independent Ethics Committee Name 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. Address of Institutional Review Board |Clinical Laboratory Facility-1:(attach separate sheet for each facility, if necessary) Aizant Drug Research Solutions Pvt. Ltd.,Diagnostics Department Name 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  Address of Clinical Laboratory Facility +91 40 23792190/91/92 Phone Number of Clinical Laboratory Facility |Clinical Laboratory Facility-2:(attach separate sheet for each facility, if necessary) Lucid Medical Diagnostics Pvt. Ltd., Name Plot No.: 203, Vasavi Nagar, Karkhana. Secunderabad, Telangana, India-500015 Address of Clinical Laboratory Facility +91 40 44114444 Phone Number of Clinical Laboratory Facility |Clinical Laboratory Facility-3: CENTROMED LABS PVT LTD., House No.: 11-20/7/A/2 to 5, Plot No. 8, 3rd Floor,  Chinnabalappa Complex, Near Moosarambagh Metro Station, Pillar No. A1478, Saleem Nagar Colony, Malakpet, Hyderabad-500036, India. Tel. 040-35162091, 92, 93, 94. |**Date Form Completed:**09 Jan 2024 |