mesЭкстремальная биомедицинаExtreme Medicine3033-89643033-8972Centre for Strategic Planning of the Federal Medical and Biological Agency10.47183/mes.2025-27-1-26-36mes-274Research ArticleГЛАВНАЯ ТЕМА: ИЗУЧЕНИЕ ТОКСИЧЕСКОГО ВОЗДЕЙСТВИЯ КСЕНОБИОТИКОВ И ПРОДУКТОВ ИХ БИОТРАНСФОРМАЦИИMAIN TOPIC: STUDY OF TOXIC EFFECTS OF XENOBIOTICS AND THEIR BIOTRANSFORMATION PRODUCTSНовая one-pot методика получения потенциальных метаболитов индапамида путем окисления-конъюгации на МАЛДИ-мишениA new one-pot technique for obtaining potential indapamide metabolites by oxidation and conjugation on MALDI targethttps://orcid.org/0000-0002-4069-0355КельциеваО. А.KeltsievaO. A.

Кельциева Ольга Александровна

Санкт-Петербург

Olga A. Keltsieva

St. Petersburg

keltcieva@gmail.comАфанасьеваА. А.AfanasyevaA. A.

Афанасьева Анна Андреевна

Санкт-Петербург

Anna A. Afanasyeva

St. Petersburg

afanasyeva.a.a.2000@yandex.ruhttps://orcid.org/0000-0003-1672-4583ИльюшонокС. К.IlyushonokS. K.

Ильюшонок Семен Кириллович

Санкт-Петербург, Ленинградская обл.

Semyon K. Ilyushonok

St. Petersburg, Leningrad region

ilsemen21@mail.ruhttps://orcid.org/0000-0002-4411-2069ГладчукА. С.GladchukA. S.

Гладчук Алексей Сергеевич - канд. техн. наук

Санкт-Петербург, Ленинградская обл.

Alexey S. Gladchuk

St. Petersburg, Leningrad region

aleglad24@gmail.comАрсеньевА. Н.ArsenievA. N.

Арсеньев Александр Николаевич

Санкт-Петербург

Alexander N. Arseniev

St. Petersburg

star2361@mail.ruФроловА. С.FrolovA. S.

Фролов Александр Станиславович

Санкт-Петербург

Alexander S. Frolov

St. Petersburg

Alexander.S.Frolov@gmail.comБабаковВ. Н.BabakovV. N.

Бабаков Владимир Николаевич - канд. биол. наук

Ленинградская обл.

Vladimir N. Babakov

Leningrad region

vbabakov@gmail.comhttps://orcid.org/0000-0003-1503-2243КрасновК. А.KrasnovK. A.

Константин Андреевич Краснов - д-р хим. наук

Санкт-Петербург

Konstantin A. Krasnov

St. Petersburg

krasnov_tox@mail.ruПодольскаяЕ. П.PodolskayaE. P.

Подольская Екатерина Петровна - д-р техн. наук

Санкт-Петербург

Ekaterina P. Podolskaya

St. Petersburg

ek.podolskaya@gmail.comНаучно-клинический центр токсикологии имени академика С. Н. Голикова Федерального медико-биологического агентства; Институт аналитического приборостроения Российской академии наукРоссияGolikov Research Center of Toxicology; Institute of Analytical Instrumentation RASRussian FederationНаучно-клинический центр токсикологии имени академика С. Н. Голикова Федерального медико-биологического агентстваРоссияGolikov Research Center of ToxicologyRussian FederationНаучно-клинический центр токсикологии имени академика С. Н. Голикова Федерального медико-биологического агентства; Научно-исследовательский институт гигиены, профпатологии и экологии человека Федерального медико-биологического агентстваРоссияInstitute of Analytical Instrumentation RAS; Research Institute of Hygiene, Occupational Pathology and Human EcologyRussian FederationНаучно-клинический центр токсикологии имени академика С. Н. Голикова Федерального медико-биологического агентства; Институт аналитического приборостроения Российской академии наук; Научно-исследовательский институт гигиены, профпатологии и экологии человека Федерального медико-биологического агентстваРоссияGolikov Research Center of Toxicology; Institute of Analytical Instrumentation RAS; Research Institute of Hygiene, Occupational Pathology and Human EcologyRussian FederationИнститут аналитического приборостроения Российской академии наукРоссияInstitute of Analytical Instrumentation RASRussian FederationНаучно-исследовательский институт гигиены, профпатологии и экологии человека Федерального медико-биологического агентстваРоссияResearch Institute of Hygiene, Occupational Pathology and Human EcologyRussian Federation2025220320252712636Copyright © Кельциева О.А., Афанасьева А.А., Ильюшонок С.К., Гладчук А.С., Арсеньев А.Н., Фролов А.С., Бабаков В.Н., Краснов К.А., Подольская Е.П., 20252025Кельциева О.А., Афанасьева А.А., Ильюшонок С.К., Гладчук А.С., Арсеньев А.Н., Фролов А.С., Бабаков В.Н., Краснов К.А., Подольская Е.П.Keltsieva O.A., Afanasyeva A.A., Ilyushonok S.K., Gladchuk A.S., Arseniev A.N., Frolov A.S., Babakov V.N., Krasnov K.A., Podolskaya E.P.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.extrememedicine.ru/jour/article/view/274Введение

Введение. Метаболическая активация ксенобиотиков, в том числе лекарственных средств, считается одним из основных механизмов развития идиосинкразических реакций. Соответственно, потенциальная биоактивация ксенобиотика должна быть тщательно оценена на ранних этапах разработки лекарств. В связи с этим поиск новых быстрых и эффективных методик скрининга реакционноспособных метаболитов ксенобиотиков является актуальным.

Цель

Цель. Разработка новой методики моделирования процессов биотрансформации ксенобиотиков in vitro для выявления потенциальных метаболитов индапамида.

Материалы и методы

Материалы и методы. В качестве методов сравнения были выбраны такие неферментативные инструментальные методы, как электрохимическое окисление (ЭХО) и фотокаталитическое окисление (ФКО) в объеме. Моделирование второй фазы метаболизма осуществлялось путем инкубации продуктов окисления индапамида с улавливающим агентом (глутатион, GSH). Продукты окисления, а также их конъюгаты с GSH затем анализировали методом высокоэффективной жидкостной хроматографии с тандемной масс-спектрометрией. В основе новой one-pot методики моделирования метаболизма лежит проведение УФ-индуцированного ФКО ксенобиотика в присутствии GSH на функционализированной диоксидом титана поверхности мишени с последующей регистрацией продуктов с помощью масс-спектрометрии с матрично-активированной лазерной десорбцией/ионизацией.

Результаты

Результаты. В случае ЭХО было обнаружено 5 метаболитов и 3 аддукта с GSH, а при использовании ФКО в объеме было выявлено 7 метаболитов и 1 аддукт с GSH. При использовании новой one-pot методики было найдено 8 аддуктов с GSH. Помимо ряда выявленных известных метаболитов индапамида и их конъюгатов с GSH, в совокупности тремя методами для индапамида было зафиксировано по 4 ранее не изученных метаболита и аддукта с GSH.

Выводы

Выводы. По сравнению с ЭХО и ФКО в объеме предложенный аналитический подход к моделированию метаболизма индапамида показал более высокую информативность в сочетании с простотой и экспрессностью, что делает его перспективным для использования в доклинических исследованиях лекарственных препаратов при прогнозировании метаболизма и токсичности объектов фармацевтической разработки, а также при изучении процессов биотрансформации различных ксенобиотиков.

Introduction

Introduction. Metabolic activation of xenobiotics, including pharma drugs, is considered to be one of the main mechanisms for the development of idiosyncratic reactions. Accordingly, the potential bioactivation of a xenobiotic should be carefully evaluated in the early stages of drug development. In this regard, the search for new rapid and effective screening techniques for reactive metabolites of xenobiotics presents particular interest.

Objective

Objective. Development of a new technique for modeling the processes of xenobiotic biotransformation in vitro to identify potential metabolites of indapamide.

Materials and methods

Materials and methods. Non-enzymatic instrumental methods, such as electrochemical oxidation (ECO) and photocatalytic oxidation (PCO) in volume, were used as comparison methods. The second phase of metabolism was modeled by incubating the oxidation products of indapamide with a trapping agent (glutathione, GSH). The oxidation products, as well as their conjugates with GSH, were then analyzed by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS). The developed one-pot technique for metabolism modeling is based on a UV-induced PCO of a xenobiotic in the presence of GSH on the surface of a target functionalized with titanium dioxide followed by detection of the products by matrix-assisted laser desorption/ionization mass spectrometry (MALDI).

Results

Results. In use of ECO resulted in the detection of 5 metabolites and 3 adducts with GSH, while the use of PCO in the volume allowed detection of 7 metabolites and 1 adduct with GSH. The new one-pot technique detected 8 adducts with GSH. In addition to the detection of a number of known indapamide metabolites and their conjugates with GSH, a total of 4 previously unstudied metabolites and adducts with GSH were each detected for indapamide by the three methods.

Conclusions

Conclusions. In comparison with ECO and PCO in volume, the proposed analytical technique for modeling indapamide metabolism showed its higher informativity combined with simplicity and rapidity, which makes it a promising candidate for use in preclinical studies of drugs in predicting the metabolism and toxicity of pharmaceutical objects, as well as in studying the biotransformation processes of various xenobiotics.

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