Russian Journal of BiotherapyRussian Journal of BiotherapyРоссийский биотерапевтический журнал1726-97841726-9792Publishing House ABV Press135810.17650/1726-9784-2022-21-4-62-70ORIGINAL REPORTSОРИГИНАЛЬНЫЕ СТАТЬИCytotoxic and antitumor properties of methionine γ-lyase conjugate in combination with S-alk(en)yl–L-cysteine sulfoxidesЦитотоксические и противоопухолевые свойства конъюгата метионин γ-лиаза-дайдзеин в комбинации с сульфоксидами S-алк(ен)ил-L-цистеинаhttps://orcid.org/0000-0001-5391-5077Abo QouraL.Або КураЛ.
Russian Federation

6 Miklukho-Maklaya St., 117198 Moscow

117198 Москва, ул. Миклухо-Маклая, 6

https://orcid.org/0000-0003-2922-2793MorozovaE. A.МорозоваЕ. А.
Russian Federation

32 Vavilova St., 119991 Moscow

119991 Москва, ул. Вавилова, 32

KovalV. S.КовальВ. С.
Russian Federation

32 Vavilova St., 119991 Moscow

119991 Москва, ул. Вавилова, 32

https://orcid.org/0000-0003-0618-1407KulikovaV. V.КуликоваВ. В.
Russian Federation

32 Vavilova St., 119991 Moscow

119991 Москва, ул. Вавилова, 32

SpirinaT. S.СпиринаТ. С.
Russian Federation

24 Kashirskoe Shosse, 115522 Moscow

115552 Москва, Каширское шоссе, 24

DemidovaE. A.ДемидоваЕ. А.
Russian Federation

24 Kashirskoe Shosse, 115522 Moscow

115552 Москва, Каширское шоссе, 24

DemidkinaT. V.ДемидкинаТ. В.
Russian Federation

32 Vavilova St., 119991 Moscow

119991 Москва, ул. Вавилова, 32

https://orcid.org/0000-0003-4006-9320PokrovskyV. S.ПокровскийВ. С.
Russian Federation

Vadim Sergeevich Pokrovsky 

6 Miklukho-Maklaya St., 117198 Moscow

24 Kashirskoe Shosse, 115522 Moscow

1 Olympiyskiy Ave., 354340 Sochi

Вадим Сергеевич Покровский 

117198 Москва, ул. Миклухо-Маклая, 6

115552 Москва, Каширское шоссе, 24

354340 Сочи, Олимпийский пр-т, 1

v.pokrovsky@ronc.ruResearch Institute of Molecular and Cellular Medicine, RUDN UniversityНИИ молекулярной и клеточной медицины ФГАОУ ВО «Российский университет дружбы народов»V.A. Engelgardt Institute of Molecular Biology of the Russian Academy of SciencesФГБУН «Институт молекулярной биологии им. В.А. Энгельгардта» Российской академии наукN.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of RussiaФГБУ «Национальный медицинский исследовательский центр онкологии им. Н.Н. Блохина» Минздрава РоссииScientific Center for Translational Medicine, Sirius University of Science and TechnologyНаучный центр трансляционной медицины АНО ВО «Научно-технологический университет «Сириус»1012202221462701012202210122022https://bioterapevt.abvpress.ru/jour/article/view/1358

Background. Enzyme prodrug therapy is a promising strategy to treat solid malignancies. The utilization of two-component systems, including an enzyme and a non-toxic prodrug, to generate cytotoxic compounds directly at the surface of the tumor cell can be successful strategy in reducing the overall toxic load on the body.

Aim. To determine antitumor activity of the pharmacological pair C115H methionine γ-lyase (C115H MGL) conjugated with daidzein (C115H MGL-Dz) and of S-alk(en)yl-L-cysteine sulfoxides against various types of solid tumors in vitro and in vivo.

Materials. MTT-test was used to determine the cytotoxicity of C115H MGL-Dz in the presence of S-alk(en)yl-L-cysteine sulfoxides in vitro against Sw620 (colon cancer), Panc1 (pancreatic cancer), and 22Rv1 (prostate cancer). Apopto- sis induction and cell cycle alteration in 22Rv1, Sw620, and SKBR3 cell lines were studied using the Muse® Caspase-3/7 and Muse® Cell Cycle Assay kits. In vivo anticancer activity was studied on Sw620, Panc1, and 22Rv1 subcutaneous xenografts in Balb/c nude mice.

Results. The C115H MGL-Dz had the maximum cytotoxic activity in the presence of S-propyl-L-cysteine sulfoxide (propiin) with IC50 values: 3.88 and 5.4 for Panc1 and 22Rv1, respectively. Dipropyl thiosulfinate formed by the β-eli-mination of propiin catalyzed by C115H MGL-Dz, induces apoptosis through both the activation of caspases and alternative pathways, and also it inhibits cell division, contributing to a decrease in the concentration of cells in the G2/M phase. The anticancer efficacy of the pharmacological pair C115H-Dz/propiin in vivo indicated a significant decrease in Panc1 tumor volume (tumor growth inhibition (TGI) 67.5 %, p = 0.004), Sw620 (TGI 22.07 %, p = 0.12) and 22Rv1 (TGI 70 %, p = 0.043).

Conclusion. Pharmacological pair C115H MGL-Dz/propiin was capable of suppressing tumor development in malignant solid tumors and might be considered as a potential anticancer approach in cancer prodrug therapy.

Введение. Ферментная пролекарственная терапия является новой перспективной стратегией в лечении солидных злокачественных новообразований. Использование двухкомпонентных систем, содержащих фермент и нетоксичный субстрат (пролекарство), позволяет вырабатывать цитотоксические вещества непосредственно у поверхности опухолевой клетки, тем самым снижая общую токсическую нагрузку на организм.

Цель исследования – оценка противоопухолевой активности фармакологических пар C115H метионин γ-лиазы (С115Н МГЛ), конъюгированной с дайдзеином (С115Н МГЛ-Dz), и сульфоксидов S-алк(ен)ил-L-цистеина в отношении солидных опухолей in vitro и in vivo.

Материалы и методы. Для определения цитотоксичности фармакологических пар in vitro на клеточных линиях рака толстой кишки (Sw620), поджелудочной железы (Panc1) и предстательной железы (22Rv1) использовали МТТ-тест. Индукцию апоптоза и изменение клеточного цикла клеток линий 22Rv1, Sw620 и SKBR3 исследовали с помощью набора реагентов Muse® Caspase-3/7 и Muse® Cell Cycle Assay kit. Противоопухолевую активность in vivo оценивали на ксенографтах Sw620, Panc1 и 22Rv1 у мышей Balb/c nude.

Результаты. Показано, что конъюгат С115Н МГЛ-Dz обладает максимальной цитотоксичностью в присутствии сульфоксида S-пропил-L-цистеина (пропиина) с IC50, равными 3,88 и 5,4 для Panc1 и 22Rv1 соответственно. Дипропилтиосульфинат, образующийся в результате реакции β-элиминирования пропиина, катализируемой C115H МГЛ-Dz, индуцирует апоптоз как посредством активации каспаз, так и через альтернативные пути, а так- же угнетает клеточное деление, способствуя снижению концентрации клеток в фазе G2/M. Исследование противоопухолевой активности фармакологической пары in vivo показало уменьшение объема опухоли у ксенографтов Panc1 (торможение роста опухоли (ТРО) 67,5 %, p = 0,004), Sw620 (ТРО 22,07 %, p = 0,12) и 22Rv1 (ТРО 70 %, p = 0,043).

Выводы. Фармакологическая пара С115Н МГЛ-Dz/пропиин значительно уменьшает рост злокачественных солидных опухолей и может рассматриваться в качестве возможного терапевтического агента в ферментной пролекарственной терапии злокачественных новообразований.

methionine γ-lyaseenzyme prodrug therapydrug targetingsubstrate-prodrugthiosulfinatesметионин γ-лиазаферментная пролекарственная терапиянаправленная доставкасубстраты-пролекарстватиосульфинатыThe authors thank N.Yu. Anisimova, PhD, DSc, a leading researcher at the Research Institute for Experimental Diagnostics and Therapy of Tumors of the N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia, for assistance in conducting experiments in vitro. In vitro and in vivo experiments were performed with the financial support by the Ministry of Science and Higher Education of the Russian Federation (Agreement on subsidies No. 075-15-2021-1060 dated on September 28, 2021). The synthesis of daidzein and conjugate C115H MGL-Dz was supported by the Russian Science Foundation (project No. 20-14-00258).Авторы благодарят д.б.н. Н.Ю. 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