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

Abstract

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 %).