Sacubitril 97 mg and Valsartan 103 mg Film-coated Tablets Relative to Entresto 97 mg/103 mg Film-coated Tablets

Title A Bioequivalence study of a randomized, open-label, single oral dose, full replicated crossover design with four-period, two-treatment, and two-sequence of Sacubitril 97 mg and Valsartan 103 mg film-coated tablets relative to Entresto 97 mg/103 mg film-coated tablets in healthy Thai volunteers under fasting condition.

Objectives The primary objective is to compare the rate and extent of absorption of a generic formulation with that of a reference formulation when given as equal labeled dose. The secondary objective is to evaluate the safety after oral administration of both test and reference formulation in healthy Thai volunteers.

Study Design Randomized, open-label, single dose, fully replicate crossover design with four-period, two-treatment and two-sequence under fasting condition and at least 7 days washout period between the doses.

Sample Size 40 Healthy Human Thai subjects. Four extra subjects if available, may be checked-in on the day of check in of period-I to compensate for any dropout prior to dosing of period-I. These subjects will be dosed if there are dropouts prior to dosing in period-I. If there are no dropouts, these subjects will be checked-out without being dosed after completion of dosing in period-I.

Drug-Product Test-Product: Sacubitril 97 mg and Valsartan 103 mg film-coated tablets Reference-product: ENTRESTOTM (Film-Coated Tablets) 97/103 mg Manufactured by: Novartis Singapore Pharmaceutical Manufacturing Pte. Ltd., Singapore for Novartis Pharma AG, Basle, Switzerland

Administration After an overnight fasting at clinical facility of at least 10 hours, each volunteer will receive a single dose of Sacubitril 97 mg and Valsartan 103 mg film-coated tablets of either test or reference with 250 mL of drinking water. Each volunteer will be allowed to drink water as desire except 1 hour before and after drug administration. The formulation is given in a full replication crossover fashion as per the randomization schedule. After the administration, the subject's oral cavity will be checked by using flashlight to confirm complete medication and fluid consumption by pharmacist.

Blood Schedule In each period, a total of 24 blood samples (approximately 7 mL each) will be collected pre-dose (0.000 hour) and at 0.083, 0.167, 0.333, 0.500, 0.670, 0.830, 1.000, 1.330, 1.670, 2.000, 2.500, 3.000, 3.500, 4.000, 5.000, 6.000, 8.000, 10.000, 12.000, 16.000, 24.000, 36.000 and 48.000 hours after study drug administration, respectively. The sample collection at 36.000 and 48.000 hours after dosing will be on ambulatory basis (i.e. on separate visit).

Sample Collection Blood samples will be collected through an indwelling catheter placed in a vein using disposable syringe or through fresh venipuncture with disposable syringes and needles. Approximately 7 mL blood sample will be withdrawn and transferred to sample collection pre-labeled tubes containing K3EDTA as anticoagulant at each sampling time point. After collection of blood samples from each subject at each time point, samples will be centrifuged at 4000 rpm for 5 minutes at 4±2°C. After centrifugation, the plasma samples will be aliquot into four pre-labeled cryovials, separating into two for sacubitril and two for valsartan (with "O" Original and "D" Duplicate label). The original cryovials for each molecule will be filled with approximately 1 mL of plasma, leaving the remaining volume for the duplicate cryovials. Cryovials containing plasma sample will be stored at -70±10 °C.

Analytical Method Sacubitril and Valsartan plasma concentration will be assayed as per Thai TFDA requirements/ international Guidelines/In-house SOP by using a UPLC-MS/MS method

Pharmacokinetic Parameters Primary pharmacokinetic parameter: Cmax, AUC0→t and AUC0→∞ and secondary pharmacokinetic parameter: Tmax, T1/2, Kel, AUC0→t/AUC0→∞ will be determined from the plasma concentration data of analytes.

Statistical Analysis ANOVA, two one-sided tests for bioequivalence, for log-transformed pharmacokinetic parameters Cmax, AUC0→t and AUC0→∞ will be performed.

Acceptance Criteria for Bioequivalence To be considered as bioequivalent For AUC: The 90% CI for the ratio of AUC0→t and AUC0→∞ of Sacubitril and Valsartan from test and reference products on the log-transformed data should be between 80.00-125.00%.

For Cmax:

The 90% CI for the ratio of Cmax of Sacubitril and Valsartan from test and reference products on the log-transformed data should be in the acceptance criteria based on the within-subject variability of reference product (ISCV) seen in the study using scaled-average-bioequivalence as recommended in the 2010 EMA guideline. if ISCV>30% for reference product in the study, then

1. 90% Confidence Interval for the ratio of Cmax should be within widened acceptance range**
2. The geometric mean ratio (GMR) of Test and Reference should be 80.00-125.00% if ISCV<30% for reference product in the study, then 90% Confidence Interval for the ratio of Cmax should be within 80.00-125.00%

• Widening of Acceptance Range for Cmax:

The extent of the widening is defined based upon the within-subject variability obtained in the bioequivalence study using scaled-average-bioequivalence according to [U, L] = exp [±k·SWR], where U is the upper limit of the acceptance range, L is the lower limit of the acceptance range, k is the regulatory constant set to 0.760 and SWR is the within-subject standard deviation of the log-transformed values of Cmax of the Reference product.

Acceptance criteria for Cmax can be widened to a maximum of 69.84 - 143.19%, if within-subject variability of reference product for Ln- transformed Cmax ≥ 50%