РЕЗУЛЬТАТЫ ЭКСПЕРИМЕНТОВ ПО РАЗРАБОТКЕ ВЫСОКООЧИЩЕННОЙ ГИАЛУРОНАТ-ЭНДО-β-N-АЦЕТИЛГЕКСОЗАМИНИДАЗЫ В КАЧЕСТВЕ ФАРМАЦЕВТИЧЕСКОЙ СУБСТАНЦИИ ДЛЯ ЛЕКАРСТВЕННОГО ПРЕПАРАТА

Abstract

Enzyme preparations, including hyaluronidase, are widely used in medical practice. Most drugs with this active substance on the pharmaceutical market are produced mainly from testicular hyaluronidase, the raw material for which are bovine testes. The technologies of enzyme preparation production are quite well known and diverse. However, the creation of a different method will allow the highly purified hyaluronate-endo-β-N-acetylhexosaminidase with high specific enzymatic activity. Objective: develop a technology and obtain a laboratory batch of the highly purified pharmaceutical substance hyaluronate-endo-β-N-acetylhexosaminidase. Materials and Methods. In the experimental study, preparations of bovine testes extracts were taken to obtain highly purified hyaluronate-endo-β-N-acetylhexosaminidase. In the first stage the protein spectrum of the testes extracts was studied, proteins were fractionally precipitated, and substances of different chemical nature were fractionated by chromatography methods. At the second stage, the development of a method to obtain the pharmaceutical substance hyaluronate-endo-β-N-acetylhexosaminidase was carried out using the methods described further in the work. Results. Based on the results obtained during the experimental study, P-II, a preparation of bovine testes extract produced by Samson-Med LLC (St. Petersburg), was chosen as the initial substance of crude hyaluronidase for further studies. The most optimal is the purification scheme, which includes as the main method affinity chromatography on heparin-sepharose using affinity elution with heparin solution in buffer with a concentration of 3 mg/ml. Conclusions As a result of the experimental study, the optimal scheme was presented to obtain the pharmaceutical substance HEBNA with high enzymatic total activity of 289 (unit of action) was presented, and a laboratory batch of highly purified hyaluronate-endo-β-N-acetylhexosaminidase was obtained.