The first generic tirzepatide GP30931: physicochemical and biological similarity to the reference drug
I.A. Lugovik, A. V. Babina, Sergey Arutyunyan, D. О. Ermolaeva, В. Б. Сапарова, T. N. Kobeleva, P. G. Zaikin, Anna D. Mikushina, Anton Bukatin, Igor E. Eliseev, B. Ts. Zajchik, Y. S. Keruchenko, I.S. Okhrimenko, Eduard V. Bocharov, Amir Taldaev, Владислав А. Лушпа, И. Е. Макаренко, Р. В. Драй
Abstract
Introduction. Metabolic syndrome is a multifactorial complex of metabolic disorders accompanied by type 2 diabetes mellitus and abdominal obesity. In the pharmacological correction of obesity and type 2 diabetes mellitus, the greatest effect is demonstrated by dual agonists of glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP), in particular tirzepatide. The original drug (Mundjaro, pln: tirzepatide) is not registered in the Eurasian Economic Union (EAEU), which limits its availability to patients and leads to the need to develop a domestic generic. Aim. Conducting a physicochemical and biological characterization of the drug Sejaro® (GP30931) in comparison with the original drug Mundjaro® to establish their comparability. Materials and methods. Three batches of each drug (solution for subcutaneous administration, 25 mg/ml) were studied. The primary structure was verified using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), peptide mapping and NMR (2D-NMR- 1 H- 1 H 1 H/ 1 H-TOCSY). Secondary structure, higher order structures and aggregation were assessed by circular dichroism (CD), high-resolution nuclear magnetic resonance (NMR), fluorescence quantum yield (FQY) and dynamic light scattering (DLS). Impurity profiles were analyzed using size-exclusion and reversed-phase HPLC. Functional activity was determined in cellular tests for activation of calcium currents through GLP-1 and GIP receptors with an assessment of the half-maximal effective concentration (EC 50 ); equivalence was confirmed by the two-sided test (TOST, 90 % confidence interval). Results and discussion. HPLC-MS and mapping confirmed identical mass (4,810.52 Da) and amino acid sequence. NMR spectra, CD and CF profiles, and DLS were completely consistent, which allows us to conclude that the secondary and tertiary structures of the preparations are structurally identical. The content of all types of impurities in GP30931 did not exceed the Munjaro values. The difference in EC 50 values for activation of GLP-1 and GIP receptors was ≤5 %, the comparative biological activity was within 0.80–1.25. Conclusion. The complex of studies conducted using orthogonal methods showed that "Sejaro" (GP30931) and "Mundjaro" are structurally identical, comparable in impurity profile and biologically equivalent. The obtained data make a significant contribution to the scientific substantiation of the equivalence of the reproduced drug tirzepatide and the reference original drug, thereby providing a reliable basis for further registration and clinical use of Sejaro® without conducting additional clinical studies.