Study to Compare the Pharmacokinetics, Pharmacodynamics, Safety, Tolerability, and Immunogenicity of Biosimilar Denosumab With Prolia® in Healthy Adult Male Volunteers

A Randomized, Double-blind, Three-arm, Parallel-group, Single-dose Study to Compare the Pharmacokinetics, Pharmacodynamics, Safety, Tolerability, and Immunogenicity of Denosumab (ENZ215, EU-sourced Prolia®, and US-sourced Prolia®) in Healthy Adult Male Volunteers

This is a randomized, double-blind, three-arm, parallel-group, single-dose study to demonstrate bioequivalence of ENZ215 and EU- and US-sourced Prolia after a single 60-mg dose administered subcutaneously in healthy adult male volunteers.

Study Overview

• Status: Completed
• Conditions: Healthy Male Subjects
• Intervention / Treatment: Biological: ENZ215; Biological: EU Sourced Prolia; Biological: US Sourced Prolia

Detailed Description

Approximately 207 subjects will be enrolled into 3 groups (69 in each group) in parallel. The subjects may be enrolled in multiple groups at the site. All eligible subjects will be assigned to one of the three treatment groups in 1:1:1 ratio i.e. ENZ215 or US-sourced Prolia® or EU-sourced Prolia® to enter into the study period of 39 weeks. The study duration will be approximately 16 months (i.e. 6 months of recruitment period, 4 weeks of screening period and approximately 39 weeks (270 days) of study period).

Each subject will be required to visit the site for a total of 20 visits: visit 1 - screening visit, visit 2 - day 0 to day 2, visit 3 - day 3, visit 4 - day 4, visit 5 - day 5, visit 6 - day 6, visit 7 - day 8, visit 8 - day 10, visit 9 - day 12, visit 10 - day 16, visit 11 - day 21, visit 12 - day 28 (week 4), visit 13 - day 42 (week 6), visit 14 - day 63 (week 9), visit 15 - day 90 (week 13), visit 16 - day 119 (week 17), visit 17 - day 147 (week 21), visit 18 - day 180 (week 26), visit 19 day - 224 (week 32), and visit 20 - day 270 (week 39). A window period of ±1 day is allowed for visit 12 (day 28), window period of ±3 days are allowed from day 42 (week 6) to day 180 (week 26), A window period of ±5 days are allowed from day 224 (week 32) to day 270 (week 39).

End of Study Assessment will be performed on day 270 (week 39) or at the time of early discontinuation of the subject.

• Study Type: Interventional
• Enrollment (Actual): 207
• Phase: Phase 1

Contacts and Locations

Study Locations

• Bulgaria
  • Sofia City Province
    • Sofia, Sofia City Province, Bulgaria, 1000
      • MC Comac Medical
• Poland
  • Warsaw, Poland, 02-172
    • MTZ Clinical Research powered by Pratia, Pratia S.A

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 28 years to 55 years (Adult)
• Accepts Healthy Volunteers: Yes

Description

INCLUSION CRITERIA

The subjects will be included in the study based on the following criteria:

1. Able to understand and give written, voluntary informed consent for the study
2. Healthy adult male volunteers between 28 to 55 years of age (both inclusive)
3. Body Mass Index (BMI) ≥ 18.50 and ≤ 30.00 kg/m2 at the time of screening
4. Medically healthy with no clinically significant medical history, vital signs, physical examination, and laboratory profiles
5. Normal or clinically acceptable 12-lead electrocardiogram, QT interval corrected for heart rate (QTc interval)* ≤ 450 msec at the time of screening
6. Subjects with negative alcohol test (breath analyzer or any suitable test) at the time of screening and admission (pre-dose)
7. Male subjects with female partners who agree to use effective contraception during study#
8. Male subjects who agree not to donate sperm during study
9. Willing and able to comply with the protocol requirements

10. Willing for multiple sampling and admission at the phase 1 study site day before dosing.

    • Note: QTc interval will be calculated using the Bazette and Fridericia formula.

      • Effective contraception: A non-vasectomised Male volunteers with female partners of child bearing potential should use dual method of contraception i.e. condom with spermicide method of contraception.

Female partners should use hormonal or non-hormonal method of contraception. (No restrictions are required for a vasectomised male provided his vasectomy has been performed 4 months or more prior to the first dosing. A male who has been vasectomised less than 4 months prior to the first dosing must follow the same restrictions as a non-vasectomised male).

EXCLUSION CRITERIA

The subjects will be excluded from the study based on the following criteria:

1. Known hypersensitivity to Denosumab or to any of the components of the study drug

2. Participating or has received any investigational drug (or is currently using an investigational device) within 30 days before receiving the study drug, or at least 10 times the respective elimination halflife (whichever period is longer) *

   * For monoclonal antibody refer exclusion criteria number 18 and 19

3. A serious infection (associated with housing and/or required intravenous anti-infectives) within 6 months before study drug administration and/or any active infection within 4 weeks of screening requiring oral or systemic antibiotics
4. History of significant drug abuse within 12 months before screening or a use of soft drugs (such as marijuana) within 3 months before the screening visit or hard drugs (such as cocaine, phencyclidine, and crack etc.) within 12 months before screening
5. Smokers who smoke ≥ 10 cigarettes or equivalent per day within 90 days prior to screening
6. Subjects with positive urine screen for drugs of abuse at the time of screening or check-in
7. Subjects with Urine Cotinine > 500ng/ml at the time of screening or check-in
8. Subjects with risk of osteonecrosis of the jaw i.e. poor oral hygiene, periodontal disease, poorly fitting dentures, history of dental disease or have undergone invasive dental procedures e.g. tooth extractions within last 6 months prior to screening.
9. Subjects with a predictable risk of invasive dental surgery during the 9 months after dosing or with planned invasive dental procedure
10. Subjects with known bone disease or recent fracture or abnormalities of calcium metabolism
11. Loss of blood (excluding volume drawn at screening) of 50 mL to 499 mL within 30 days, or more than 499 mL within 56 days before dosing
12. History of immunodeficiency (including those subjects with a positive test for human immunodeficiency virus [HIV] at screening)
13. Have a positive result for hepatitis B antigen test (HBsAg) or hepatitis C antibody test (HCAb), or show evidence of possible infection
14. Major surgical procedure within 28 days of dose of investigational product.
15. Male subjects having pregnant female partner at the time of screening.
16. Subject with a history of recurrent or chronic infections
17. Received live vaccines within 4 weeks or who may require live vaccine(s) during the study duration
18. Prior use of denosumab
19. Have previously been exposed to a monoclonal antibody or fusion protein within 270 days (other than denosumab) prior to randomisation and/or there is confirmed evidence or clinical suspicion of immunogenicity from previous exposure to a monoclonal antibody or fusion protein.
20. Any reason/condition which would preclude subject's participation in the study as per the Investigator's opinion or warnings and contraindications in the prescribing information of Prolia
21. Subjects with suspected signs and symptoms of COVID-19/confirmed novel coronavirus infection (COVID-19).

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Double

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: ENZ215; ENZ215 Injection:- 60 mg Denosumab (ENZ215) will be administered subcutaneously on day 1.	Biological: ENZ215; healthy volunteers receive ENZ215 (60mg) once
Active Comparator: EU Sourced Prolia; EU sourced Prolia Injection:- 60 mg Denosumab (EU sourced Prolia) will be administered subcutaneously on day 1.	Biological: EU Sourced Prolia; healthy volunteers receive Denosumab (60mg) once
Active Comparator: US Sourced Prolia; US sourced Prolia Injection:- 60 mg Denosumab (US sourced Prolia) will be administered subcutaneously on day 1.	Biological: US Sourced Prolia; healthy volunteers receive Denosumab (60mg) once

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Maximum Observed Drug Concentration (Cmax) of ENZ215 and EU- and US-sourced Prolia®	A total of twenty four blood PK samples of 2.5 mL each were collected from each subject in the study. Pre-dose sample were collected within 30 minutes prior to IP administration. Post-dose samples up to Day 2 were collected within ± 10 minutes, within ± 2 hours from Day 3 to Day 21, within ± 1 day on Day 28, within ± 3 days from Day 42 to Day 180 and within ±5 days from Day 224 to Day 270. Serum concentrations of denosumab were measured at central laboratory by a validated analytical method.	270 days
Area Under the Drug Concentration-time Curve From Day 0 to Day 270 (AUC0-t) of ENZ215, EU- and US-sourced Prolia®. and EU- and US-sourced Prolia®	A total of twenty four blood PK samples of 2.5 mL each were collected from each subject in the study. Pre-dose sample were collected within 30 minutes prior to IP administration. Post-dose samples up to Day 2 were collected within ± 10 minutes, within ± 2 hours from Day 3 to Day 21, within ± 1 day on Day 28, within ± 3 days from Day 42 to Day 180 and within ±5 days from Day 224 to Day 270. Serum concentrations of denosumab were measured at central laboratory by a validated analytical method.	270 days
Area Under the Drug Concentration-time Curve From Time 0 to Infinity (AUC0-inf) of ENZ215, EU- and US-sourced Prolia®. and EU- and US-sourced Prolia®	A total of twenty four blood PK samples of 2.5 mL each were collected from each subject in the study. Pre-dose sample were collected within 30 minutes prior to IP administration. Post-dose samples up to Day 2 were collected within ± 10 minutes, within ± 2 hours from Day 3 to Day 21, within ± 1 day on Day 28, within ± 3 days from Day 42 to Day 180 and within ±5 days from Day 224 to Day 270. Serum concentrations of denosumab were measured at central laboratory by a validated analytical method.	270 days

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Partial Area Under the Drug Concentration-time Curve From Time 0 (Pre-dose) to Day 28	AUC0-28 days were compared between ENZ215 and Prolia® using tests after log-transformation wherever appropriate.	28 days
Time to Reach Cmax (Tmax)	The non-parametric analysis was used for the comparison of tmax between ENZ215 and Prolia®.	270 days
Terminal Elimination Half-life (t1/2)	t1/2 was compared between ENZ215 and Prolia® using tests after log-transformation wherever appropriate.	270 days
Apparent Systemic Clearance (CL/F)	CL/F was compared between ENZ215 and Prolia® using tests after log-transformation wherever appropriate.	270 days
Area Under the Effect Curve (AUEC) From Time 0 to Day 270 for Serum CTX-1 Percent Inhibition Percent Inhibition	The AUEC was calculated as the area under the effect curve from baseline until CTX-1 values return to baseline for the first time. A total of sixteen (16) blood samples for serum CTX-1 estimation of 3.5 mL each was collected from each subject in the study. For CTX-1, blood samples were collected at the same time and after a minimum of 10 hours of fasting.	270 days

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Number of Subjects Who Developed Denosumab Neutralizing Antibodies and Antidrug Antibodies. Antibodies	A total of 10 blood Immunogenicity assessment samples of 5.0 mL for NAb and ADA were collected from each subject in the study. The frequency and percentage of positive ADA or NAb result was provided. The proportion of positive ADA or NAb in each treatment group was compared using chi-square or Fisher's exact tests. The p-value, relative risk, and corresponding 95% CI was presented.	270 days
Incidence of Adverse Events	Adverse events and SAEs as and when occurred, were properly recorded, evaluated, managed, and reported from signing informed consent till end of Study Assessment visit. All AEs were summarized using appropriate medical coding dictionary.	270 days

Collaborators and Investigators

• Sponsor: Enzene Biosciences Ltd.
• Collaborators: Alkem Laboratories Ltd
• Investigators: Principal Investigator: Dr. Nadine Abdullah, Celerion GB Ltd.

Study record dates

Study Major Dates

• Study Start (Actual): January 10, 2022
• Primary Completion (Actual): May 22, 2024
• Study Completion (Actual): May 22, 2024

Study Registration Dates

• First Submitted: January 11, 2022
• First Submitted That Met QC Criteria: February 8, 2022
• First Posted (Actual): February 18, 2022

Study Record Updates

• Last Update Posted (Estimated): December 23, 2025
• Last Update Submitted That Met QC Criteria: December 20, 2025
• Last Verified: December 1, 2025

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Physiological Effects of Drugs; Bone Density Conservation Agents; Denosumab
• Other Study ID Numbers: ALK22/ENZ215-DEN1; 2021-004177-32 (EudraCT Number)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No