Study to Determine the Comparative Pharmacodynamics of Enoxaparin Sodium Biosimilar With That From Clexane

Study Name: (SPRING) Study of Pharmacodynamics and Relative Bioavailability of an Invented National Generic Enoxaparin

An Open-Label, Single-center, Randomized, Single-Dose, Two-Way Crossover Biosimilarity Study to Determine the Comparative Pharmacodynamics of Enoxaparin Sodium Biosimilar 40mg/0.4ml with that from the Reference IMP, Clexane® (40 mg/0.4ml), Following Single-Dose Administration in Healthy Participants. Test: Enoxaparin Sodium (Enoxaparin Sodium 40mg/0.4ml) manufactured by EIPICO, Egypt. Reference: Clexane (Enoxaparin Sodium 40mg/0.4ml) manufactured by Sanofi Aventis, Egypt. Primary objective:

To assess biosimilarity between a single dose from the test product versus the reference product in healthy participants

Secondary objective:

To investigate the safety and tolerability of the formulations. This study is a randomized single-dose, two-way, two-period, two-sequence, crossover biosimilarity study with a washout period of one week after each dosing.A minimum of 21 healthy adult male and female participants from Egyptian population will be enrolled in this study, along with 5 additional participants to account for potential dropouts or withdrawal. 26 Participants plus 1-4 alternates will be admitted to the study. An alternate participant will be dosed by the same sequence as the withdrawn participant only if any participant of the first 26 Participants withdraws before the first study drug administration. Withdrawals after study drug administration will not be replaced. All participants will be healthy adults aged (21-55) years, with a BMI within the accepted range of 18.5-30 kg/m², and will meet the study's selection criteria.

This is an Open-Label, Single-center, Randomized, Single-Dose, Two-Way Crossover Biosimilarity Study to Determine the Comparative Pharmacodynamics of Enoxaparin Sodium Biosimilar with that from the Reference IMP, Clexane, Following Single-Dose Administration in Healthy Participants.

The data that will be collected include:

• Participant identification data
• Demographic data
• Smoking habits, caffeine use, medication use, alcohol consumption.
• Details of prior participation in any clinical or bioequivalence study.
• Participant medical history, physical examination
• Vital signs, laboratory results, ECG examination, drug abuse & alcohol test
• Study Drug Information: Details of the administered drug, including time of administration.
• Concomitant Medications
• Blood sampling intervals for Pharmacodynamic (PD) analysis.
• Safety Monitoring: Documentation of any adverse events (AEs) Sample Collection and Sample Processing An intravenous cannula will be inserted before the pre-dose sample and will remain until 24 hours post-dose. Blood samples will be collected through an indwelling cannula placed in a forearm vein using a disposable syringe. If difficulty occurs in blood withdrawing or Participants not feeling comfortable with cannula, cannula will be removed, and remaining blood samples will be collected through fresh vein puncture or by re-cannulation. 5 ml of blood samples will be withdrawn and transferred into sodium citrate collection tubes at each time interval. After collection of blood samples placed in a wet ice container equivalent to the approximate height of the blood in the tube till centrifugation, then after centrifugation, transferred into an ice box containing wet ice and stored at -70°±15° immediately. After the collection of blood samples from all Participants at each time interval, samples will be centrifuged at 3500 RPM for 10 minutes.

All plasma samples will be transferred into pre-labeled (Study code, Participant No., Period, Sampling time point) polypropylene tubes. The polypropylene tubes will be transferred to freezer area in an icebox containing wet ice, and the polypropylene tubes will be stored at -70°±15° freezers. Total Number of Blood Samples: 19 samples in each study period. Volume of each sample: 5 ml. Sampling Hours: pre-dose and 0.5, 1.0, 1.5 (1 h 30 min), 2.0, 2.333 (2 h 20 min), 2.667 (2 h 40 min), 3.0, 3.333 (3 h 20 min), 3.667 (3 h 40 min), 4.0, 4.5 (4 h 30 min), 5.0, 6.0, 8.0, 10.0, 12.0, 16.0, and 24.0 h after dose administration Statistical analysis of primary endpoints for the two periods will include descriptive Statistics, ANOVA, and Confidence Interval (C.I.) of enoxaparin. The equivalence of the products will be concluded if the two -one -sided T-test 90 % confidence interval for the test to reference ratio means is within 80.00 - 125.00 % for each of the ln-transformed data of the following primary endpoints: For Anti-Factor Xa: Anti-Xa Amax, and Anti-Xa AUEC0-t , & For Anti-Factor IIa: Anti-IIa Amax and Anti IIa AUE0-t and finally if there were no safety concerns and both products were well tolerated by the study Participants.

Inclusion Criteria:

• Healthy male and female participants aged 21 to 55 years at the time of the first dose administration.
• Body Mass Index (BMI) between 18.5 and 30 kg/m², and body weight of at least 45 kg.
• Participants must have vital signs within the normal range, as defined below, measured at pre-dose:
• Blood Pressure: Systolic 100-130 mmHg, Diastolic 70-90 mmHg.
• Pulse Rate: 60-100 beats per minute (bpm).
• Body Temperature: 36.1-37.2

Exclusion Criteria:

• Hypersensitivity to Enoxaparin or any of the formulation excipients
• Contraindication to enoxaparin or related group of drugs, which includes but not limited to significant bleeding disorders, thrombocytopenia, blood-clothing disorders, and increased risk of bleeding.
• History or presence of any medical condition or disease according to the opinion of the principal Investigator
• History or presence of significant alcoholism or drug abuse in the past one year.
• History or presence of heavy smoking (≥10 cigarettes or consumption of tobacco products and refusal to restrain from smoking or consumption of tobacco products for 48.00 hours before dosing until checkout of the study).
• History or presence of significant renal, hepatobiliary, or liver impairment; a medical or familial history of seizures; significant asthma, urticaria, or other allergic reactions; or any other significant medical condition as determined by the Principal Investigator or their delegate.
• History of difficulty with donating blood or difficulty in accessibility of veins
• Use of any prescribed medication, OTC medicines or herbal medicines during the last two weeks preceding the first dosing
• Participants who have any food allergy, intolerance, restriction, or special diet during the four weeks before screening
• Participation in a drug research study within the past 2 months planned day of first dose administration
• Donation of blood in the past 2 months before screening
• Refusal to abstain from food (fasting) for at least ten hours before dosing
• Refusal to abstain from alcohol or methylxanthine-containing beverages
• Participants with a positive test for HBs-Ag, HIV-Ab, or HCV-Ab
• Participant with a positive drug abuse test in urine at screening or at the time of check-in
• Participant with abnormal baseline coagulation parameters, such as prolonged PT (if greater than 1.2 to 1.5 times the ULN of approximately 11 to 15 seconds), aPTT (if greater than 1.5 to 2.5 times the ULN of 25 to 40 seconds), or INR values (is above 1.4) outside the normal reference range.

This is to minimize bleeding risk and confounding effects on study outcomes.