Enzyme-Linked Immunosorbent Assay Method Application in the Study of Comparative Pharmacokinetics of Insulin Glargin Preparations

Aims: adaptation and validation of the ELISA method insulin glargine determination for the pharmacokinetic study, practical approval in the biosimilars clinical trial.

Materials and methods. Serum insulin glargine determination was measured using a commercial ELISA kit. All tests were run on a Personal LAB machine (Adaltis S.r.l., Rome, Italy) with test systems for measuring the concentration of insulin glargine (Invitron Ltd., United Kingdom); human insulin concentrations were measured in the samples from the study for correction of cross-reactivity. Clinical part of this study included 42 male patients aged 18–65 with diabetes mellitus type 1. This was a double-blind, randomized, crossover clamp study with wash-out period of 7–14 days. Comparisons drugs: Insulin Glargine (glargine) solution for subcutaneous administration, 100 U/ml (GEROPHARM, Russia) and Lantus® (glargine) solution for subcutaneous administration, 100 U/ml (Sanofi-Aventis Deutschland GmbH, Germany).

Results. At the stage of the method adaptation the modification of original manufacturer’s method was performed; the full validation of modified analytical method for all parameters (selectivity, specificity, precision of calibration curves, intra- and inter-batch precision and accuracy, carry-over, dilution integrity, stability of solutions, stability in biologic matrix, parallelism) in accordance with regulatory authorities requirements has been done. The primary endpoint for long-acting insulins – AUCins.0-τ was calculated. Insulin Glargine and Lantus® are equivalent based on AUCins.0-τ data (point estimation for ratio of geometric means was 99 %, the confidence intervals for the ratio of the geometric mean for AUCins.0-τ was 81.02–120.62 %, that correspond to acceptance range 80.00–125.00 %).

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