danДоклады Национальной академии наук БеларусиDoklady of the National Academy of Sciences of Belarus1561-83232524-2431The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8323-2021-65-3-309-319dan-976Research ArticleХИМИЯCHEMISTRYИнгибирование фосфолипазы А2 производными виразолаInhibition of phospholipase A2 by Virasole derivationЛитвинкоН. М.LitvinkoN. M.

Литвинко Наталья Михайловна – д-р хим наук, доцент, заведующая лабораторией

ул. Купревича, 5/2, 220141, Минск, Республика Беларусь

Litvinko Natalia M. – D. Sc. (Chemistry), Assistant Professor, Head of the Laboratory

5/2, Kuprevich Str., 220141, Minsk, Republic of Belarus

al_h@mail.ruИнститут биоорганической химии Национальной академии наук БеларусиInstitute of Bioorganic Chemistry of the National Academy of Sciences of Belarus202115072021653309319Copyright © Литвинко Н.М., 20212021Литвинко Н.М.Litvinko N.M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://doklady.belnauka.by/jour/article/view/976

Изучена кинетика гидролиза фосфатидилхолина (ФХ) под действием панкреатической фосфолипазы А2 IB (КФ 3.1.1.4, ФЛА2) в присутствии липофильного производного виразола – 1-(3-(трет–бутилдиметилсиланилокси)-4-гидрокси-5-(((4-метоксифенил)дифенилметокси)метил) тетрагидрофуран-2-ил)-1H-1,2,4-триазол-3-карбоксамида (Виразол2ЗГ), обладающего противовирусным действием. Количественно охарактеризованы обе стадии фосфолиполиза: связывания фермента (Ks) с поверхностью раздела фаз «липид–вода» и непосредственно каталитический акт (Km) с определением максимальной скорости реакции (Vmax). Обнаружено, что Виразол2ЗГ в концентрации 0,5 мкмоль/мл не влияет на величину Ks; напротив, константа Михаэлиса, Km, возрастает в 1,8 раза наряду с постоянством параметра Vmax. На основании постоянства величин Ks можно предполагать отсутствие в условиях данного эксперимента торможения распада комплекса фермент–мицелла в присутствии эффектора. Кинетические параметры реакции (увеличение величины Km и неизменность Vmax в присутствии Виразол2ЗГ) свидетельствуют в пользу умеренного конкурентного ингибирования панкреатической ФЛА2, Ki = 65 мМ, что указывает на возможность поиска биологической активности антипанкреатитного действия в ряду про-лекарств (pro-drugs) нуклеозидной природы.

Kinetics of phosphatidylcholine (PC) hydrolysis under the action of pancreatic phospholipase A2 IB, (EC 3.1.1.4, PLA2) in the presence of a lipophilic derivative of the antiviral drug Virazole 1-(3-((tert-butyldimethylsilyl)oxy)-4-hydroxy-5- (((4-methoxyphenyl)diphenylmethoxy)methyl)tetrahydrofuran-2-yl)-1H-1,2,4-triazole-3-carboxamide (Virazole2ЗГ) was studied. The both steps of phospholipolysis were quantitatively characterized: the binding of the enzyme to the lipid-water interface (Ks) and directly the catalytic act (Km) with the determination of the maximum reaction rate (Vmax). It was found that Virazole2ЗГ at a concentration of 0.5 μmol/ml does not affect the Ks value; on the contrary, the Michaelis constant, Km, increases by a factor of 1.8 along with the constancy of the parameter Vmax. Based on the constancy of the Ks values, it seems to be assumed that there is no inhibition of the disintegration of the enzyme-micelle complex in the presence of the effector under the studied reaction conditions. The kinetic parameters of the reaction (the increase in Km and the constancy of Vmax in the presence of Virazole2ЗГ) testify in favor of a moderate competitive inhibition of pancreatic PLA2, Ki = 65 mM, which indicates the possibility of searching for the biological activity of the anti-pancreatitis action in the series of pro-drugs of nucleoside nature.

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The authors declare that there are no conflicts of interest present.