Evaluation of Dose-response, Pharmacodynamic and Pharmacokinetic Bioequivalence of Filgrastim in Healthy Male Volunteers After Single and Multiple-dose Subcutaneous Administration of the Test Injectable Formulation BK0023 vs. Neupogen®

Study Overview
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Brief Summary
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Study rationale: Bio-ker has developed the new formulation of filgrastim BK0023 with the same active content as Neupogen®. BK0023 is expected to have the same tolerability profile and clinical effects as Neupogen® in controlling myelo-toxicity induced by chemotherapy given for the treatment of solid and haematological tumours. It is worth noting that the production and manufacturing procedures allow to have a reduction of drug cost thus it is likely to have pharmacoeconomic advantages. The study is aimed at investigating the pharmacodynamic equivalence and the pharmacokinetic bioequivalence of the new BK0023 injectable formulation of filgrastim 0.3 mg/mL by Bio-Ker S.r.l. vs. the comparator (Neupogen® 0.3 mg/mL, Dompé Biotec S.p.A., Italy). Healthy male subjects will receive test and reference at the doses of 2.5 and 5 µg/kg/day for 7 consecutive days and at the dose of 10 µg/kg/day for 5 consecutive days according to a randomised cross-over design. Pharmacodynamics, pharmacokinetics and safety of BK0023 injectable formulation 0.3 mg/mL and of Neupogen® 0.3 mg/mL, administered in 2 consecutive periods with a wash-out of at least 28 days elapsing between the last injection of period I and the first of period II, are compared. Study design: Single and multiple escalating dose, double-blind, randomised, two-way cross-over, pharmacodynamic and pharmacokinetic bioequivalence study.

Detailed Description
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In a first part of the study, 16 healthy male subjects will be included in each dose group and will receive test and reference product according to the cross-over design. After the end of the second period of each dose group (completion of both cross-over periods for each dose group) the pharmacodynamic and pharmacokinetic primary parameters will be calculated. The blinding will be temporarily broken by one statistician for the ad interim analysis of primary parameters that will be performed after conclusion of the treatment of 16 subjects for each dose group (results are not disclosed) with the aim to re-calculate the sample size and to conclude the study with the necessary and sufficient number of subjects for each dose regimen. The study design was chosen according to the internationally recognised guideline for pharmacokinetics studies and in accordance with the EMEA guidance on similar medicinal products containing recombinant granulocyte-colony stimulating factor (G-CSF), which is annexed to the guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issues. A clinical development plan including the study outline was also submitted to EMEA, which examined the documentation and gave advice about the present study design . A multiple-dose design was chosen instead of a single-dose design, since G-CSF pharmacokinetics is non linear due to 2 reasons: non-proportional increase with dose and time dependent non-linearity. However, pharmacodynamic equivalence and pharmacokinetic bioequivalence will be both tested after the 1st and the last dose of the multiple dose treatment. Since the effects of r-h-met-G-CSF can be directly measured as pharmacodynamic responses, i.e. the increase in the absolute neutrophil count (ANC) and the development of peripheral blood cells (CD34+ cells), both parameters will be evaluated for the equivalence testing. Dose levels of 2.5 and 5 μg/kg/day were chosen for the pharmacodynamic equivalence, since literature data show a clear dose-response relationship in terms of the pharmacodynamic parameters over this range. The 10 μg/kg/day dose was chosen to make the investigated range relevant with respect to the clinical indications and the usual praxis for Neupogen®. Moreover, the subcutaneous administration was chosen, since this administration route is the most commonly used in the clinical setting.

Official Title
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Evaluation of Dose-response, Pharmacodynamic and Pharmacokinetic Bioequivalence of Filgrastim in Healthy Male Volunteers After Single and Multiple-dose Subcutaneous Administration of the BK0023 Injectable Formulation vs. Neupogen®

Conditions
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Healthy Male Subjects

Intervention / Treatment
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* Drug: Filgrastim test
* Drug: Filgrastim reference


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: a body mass index (BMI) between 18 and 28 kg/m2, a body weight between 60 and 90 kg, good health based on medical history, physical examination, a 12-lead electrocardiogram (ECG) and routine haematology and blood chemistry tests, willingness to provide written informed consent values of leukocytes and thrombocytes had to be inside the normality range at the screening. Exclusion Criteria: intake of any concomitant medication, a history of drug, caffeine (>5 cups coffee/tea/day) or tobacco (>/=10 cigarettes/day) abuse, or alcohol consumption in excess of two drinks per day, as defined by the U.S.D.A. dietary guidelines, ascertained or presumptive hypersensitivity to the active compound or history of anaphylaxis to drugs.

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 40 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Other: Group 1: filgrastim 2.5 µg/kg/day for 7 consecutive days<br>The test investigational product is BK0023. The reference product is Neupogen® 30, by Dompé Biotec S.p.A., Italy. Both study treatments were administered according to a cross-over design in 2 subsequent periods separated by wash-out periods of at least 28 days. The sites, where the injections are to be performed, are planned as follows: st injection: upper part of the upper arm, posterior surface nd injection: upper part of the thigh rd injection: abdomen with the exception of the umbilical area th injection: upper part of the contra-lateral upper arm, posterior surface th injection: upper part of the contra-lateral thigh th injection: abdomen with the exception of the umbilical area th injection: upper part of the same upper arm, posterior surface as for the 1st injection. | Drug: Filgrastim test<br>* 0.3 mg/mL injectable solution in 1 mL vials Manufacturer (Drug Substance): Eurogenetec S.A., B-4102 Seraing, Belgium Manufacturer (Drug Product): Areta International S.r.l., I-21040 Gerenzano, Italy Batch release: Areta International S.r.l., I-21040 Gerenzano, Italy Route of administration: subcutaneous<br>* Other names: r-metHuG-CSF, BK0023;Drug: Filgrastim reference<br>* 0.3 mg/mL injectable solution in 1 mL vials Licensed owner: Dompé farmaceutici S.p.A., Milan, Italy Manufacturer (Drug Product): Hoffmann - La Roche Ltd., Basel, Switzerland and Amgen Manufacturing Limited, Puertorico. Batch release: Amgen Europe B.V., Breda, Netherlands Route of administration: subcutaneous<br>* Other names: r-metHuG-CSF, Neupogen®;|
| Other: Group 2: filgrastim 5 µg/kg/day for 7 consecutive days<br>The test investigational product is BK0023. The reference product is Neupogen® 30, by Dompé Biotec S.p.A., Italy. Both study treatments were administered according to a cross-over design in 2 subsequent periods separated by wash-out periods of at least 28 days. The sites, where the injections are to be performed, are planned as follows: st injection: upper part of the upper arm, posterior surface nd injection: upper part of the thigh rd injection: abdomen with the exception of the umbilical area th injection: upper part of the contra-lateral upper arm, posterior surface th injection: upper part of the contra-lateral thigh th injection: abdomen with the exception of the umbilical area th injection: upper part of the same upper arm, posterior surface as for the 1st injection. | Drug: Filgrastim test<br>* 0.3 mg/mL injectable solution in 1 mL vials Manufacturer (Drug Substance): Eurogenetec S.A., B-4102 Seraing, Belgium Manufacturer (Drug Product): Areta International S.r.l., I-21040 Gerenzano, Italy Batch release: Areta International S.r.l., I-21040 Gerenzano, Italy Route of administration: subcutaneous<br>* Other names: r-metHuG-CSF, BK0023;Drug: Filgrastim reference<br>* 0.3 mg/mL injectable solution in 1 mL vials Licensed owner: Dompé farmaceutici S.p.A., Milan, Italy Manufacturer (Drug Product): Hoffmann - La Roche Ltd., Basel, Switzerland and Amgen Manufacturing Limited, Puertorico. Batch release: Amgen Europe B.V., Breda, Netherlands Route of administration: subcutaneous<br>* Other names: r-metHuG-CSF, Neupogen®;|
| Other: Group 3: filgrastim 10 µg/kg/day for 5 consecutive days<br>The test investigational product is BK0023. The reference product is Neupogen® 30, by Dompé Biotec S.p.A., Italy. Both study treatments were administered according to a cross-over design in 2 subsequent periods separated by wash-out periods of at least 28 days. The sites, where the injections are to be performed, are planned as follows: st injection: upper part of the upper arm, posterior surface nd injection: upper part of the thigh rd injection: abdomen with the exception of the umbilical area th injection: upper part of the contra-lateral upper arm, posterior surface th injection: upper part of the contra-lateral thigh. | Drug: Filgrastim test<br>* 0.3 mg/mL injectable solution in 1 mL vials Manufacturer (Drug Substance): Eurogenetec S.A., B-4102 Seraing, Belgium Manufacturer (Drug Product): Areta International S.r.l., I-21040 Gerenzano, Italy Batch release: Areta International S.r.l., I-21040 Gerenzano, Italy Route of administration: subcutaneous<br>* Other names: r-metHuG-CSF, BK0023;Drug: Filgrastim reference<br>* 0.3 mg/mL injectable solution in 1 mL vials Licensed owner: Dompé farmaceutici S.p.A., Milan, Italy Manufacturer (Drug Product): Hoffmann - La Roche Ltd., Basel, Switzerland and Amgen Manufacturing Limited, Puertorico. Batch release: Amgen Europe B.V., Breda, Netherlands Route of administration: subcutaneous<br>* Other names: r-metHuG-CSF, Neupogen®;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Area under the curve of absolute neutrophil count (AUCANCday1) and maximum absolute neutrophil count (ANCday1max); AUCANCday1-10 and AUCANCday1-8; ANCday1-10max and ANCday1-8max; AUCday1 and AUCss of filgrastim in serum. | Baseline adjusted AUC of ANC on day 1 after single dose (AUCANCday1) and from day 1 to 10 (dose groups of 2.5 and 5 µg/kg/day) or from day 1 to 8 (dose group of 10 µg/kg/day) including the elimination period (AUCANCday1-10 and AUCANCday1-8) and ANCday1max, ANCday1-10max and ANCday1-8max after treatment with filgrastim 0.3 mg/mL by Bio-ker and Neupogen® AUC of serum filgrastim after both single (AUCday1) and multiple dose (AUCss) of filgrastim 0.3 mg/mL by Bio-ker and Neupogen® | Day 1 after single dose and from day 1 to 10 (dose groups of 2.5 and 5 µg/kg/day) or from day 1 to 8 (dose group of 10 µg/kg/day) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Area under the curve of CD34+ cell count (AUCCD34+) and maximum CD34+ cell count (CD34+max); maximum concentrations at steady state (Cssmax), Cmaxday1, time to achieve Cmax (Tmax), t1/2 and clearance of serum filgrastim | Baseline adjusted AUCCD34+ from day 1 to 10 (dose groups of 2.5 and 5 µg/kg/day) or from day 1 to 8 (dose group of 10 µg/kg/day) including the elimination period and CD34+max after treatment with filgrastim 0.3 mg/mL by Bio-ker and Neupogen® Cmax, Tmax t1/2 and clearance of serum filgrastim after both single (Cmaxday1, Tmaxday1, t1/2day1 and Clday1) and multiple dose (Cssmax, Tssmax, tss1/2, Clss) of filgrastim 0.3 mg/mL by Bio-ker and Neupogen® | On day 1 after single dose and from day 1 to 10 (dose groups of 2.5 and 5 µg/kg/day) or from day 1 to 8 (dose group of 10 µg/kg/day) |