Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells

Aim. To evaluate the in vitro cytotoxicity of novel spiro-substituted thiopyrano[2,3-d]thiazoles towards tumor cells of different tissue origin. Methods. Organic synthesis; spectral methods; MTT test, statistical analysis. Results. In vitro screening of the cytotoxic activity of the 5’-carboxy-7’-ar...

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Дата:	2017
Автори:	Zelisko, N.I., Finiuk, N.S., Shvets, V.M., Medvid, Yu.O., Stoika, R.S., Lesyk, R.B.
Формат:	Стаття
Мова:	English
Опубліковано:	Інститут молекулярної біології і генетики НАН України 2017
Назва видання:	Вiopolymers and Cell
Теми:	Bioorganic Chemistry
Онлайн доступ:	http://dspace.nbuv.gov.ua/handle/123456789/152984
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Цитувати:	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells / N.I. Zelisko, N.S. Finiuk, V.M. Shvets, Yu.O. Medvid, R.S. Stoika, R.B. Lesyk // Вiopolymers and Cell. — 2017. — Т. 33, № 4. — С. 282-290. — Бібліогр.: 25 назв. — англ.

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spelling	irk-123456789-1529842019-06-14T01:29:11Z Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells Zelisko, N.I. Finiuk, N.S. Shvets, V.M. Medvid, Yu.O. Stoika, R.S. Lesyk, R.B. Bioorganic Chemistry Aim. To evaluate the in vitro cytotoxicity of novel spiro-substituted thiopyrano[2,3-d]thiazoles towards tumor cells of different tissue origin. Methods. Organic synthesis; spectral methods; MTT test, statistical analysis. Results. In vitro screening of the cytotoxic activity of the 5’-carboxy-7’-aryl-1-aryl-3’,7’-dihydro-2H,2’H,5H-spiro[pyrolidin-3,6’-thiopyrano[2,3-d]thiazol]-2,2’,5-triones and N-(4-chlorophenyl)-2-[1-(4-chlorophenyl)-2,5-dioxopyrrolidin-3-ylidene]-acetamide was performed using various cancer cell lines (Jurkat human acute T-cell leukemia cell line, MCF-7 human breast adenocarcinoma cell line, Skov3 human ovarian carcinoma cells line, SK-Mel-28 human melanoma cells line, and SW-1573 human non-small-cell lung cancer cell line). The tested compounds possessed different cytotoxic action towards the studied tumor cells. Leukemia cells appeared to be more sensitive for the studied derivatives. The cytotoxic effect of the compound 2 towards Jurkat cells was shown to be dose- and time-dependent (3, 6, 24, 48 and 72 h). This compound demonstrated the cytotoxic action towards Jurkat cells as soon as in 6 h after its addition to the cultured cells (IC₅₀ = 66 μM), and its toxicity towards these cells was more prominent after 24 h treatment (IC₅₀= 40 μM). Conclusions. The panel of thiopyrano[2,3-d]thiazole derivatives was synthesized and screened for their cytotoxic activity in vitro towards tumor cells of different tissue origin. The compound 2 was found to be the most active agent with selectivity for the leukemia cells. This compound inhibits growth of the human acute T-cell leukemia cells of Jurkat line (IC₅₀ = 33.5 μM) and possesses relatively low toxicity towards the pseudo-normal mammalian cells. Мета. Вивчення цитотоксичної активності нових спіро-заміщених похідних тіопірано[2,3-d]тіазолу на пухлинні клітин різного походження. Методи. Органічний синтез; спектральні методи; МТТ тест, статистичний аналіз. Результати. Проведено дослідження іn vitro 5’-карбокси-7’-арил-1-арил-3’,7’-дигідро-2H,2’H,5H-спіро[піролідин-3,6’-тіопірано[2,3-d]тіазол]-2,2’,5-тріонів і N-(4-хлорофеніл)-2-[1-(4-хлорофеніл)-2,5-діоксопіролідин-3-іліден]-ацетаміду щодо різних ліній пухлинних клітин (Т-клітинної лейкемії Jurkat, аденокарциноми молочної залози MCF-7, карциноми яєчника Skov3, меланоми SK-Mel-28, недрібноклітинного раку легені SW-1573). Синтезовані сполуки мають різну цитотоксичну дію щодо використаних ліній пухлинних клітин. Клітини лейкемії виявилися найбільш чутливими до дії цих похідних. Цитотоксичний ефект сполуки 2 щодо Т-клітинної лейкемії Jurkat залежить від дози сполуки і тривалості її дії (3, 6, 24, 48 і 72 год). Сполука 2 володіє цитотоксичною дією на лейкемічні клітини лінії Jurkat вже через 6 год після її додавання до культури цих клітин (IC₅₀ = 66 μМ), а через 24 год її цитотоксичність зростає (IC₅₀ = 40 μМ). Висновки. Проведено скринінг нових спіро-заміщених похідних тіопірано[2,3-d]тіазолу щодо їх цитотоксичної дії на пухлинні клітини іn vitro. Сполука 2 володіє найвищою активністю у лейкемічних Т-клітинах лінії Jurkat (IC₅₀ = 33,5 мкМ), тоді як її токсичність щодо псевдо-нормальних клітин ссавців є відносно низькою. Цель. Изучение цитотоксической активности новых спиро-замещенных производных тиопирано[2,3-d]тиазола на опухолевые клетки различной этиологии. Методы. Органический синтез; спектральные методы; МТТ тест, статистический анализ. Результаты. Исследовано действие 5’-карбокси-7’-арил-1-арил-3’,7’-дигидро-2H,2’H,5H-спиро[пиролидин-3,6’-тиопирано[2,3-d]тиазол]-2,2’,5-трионов и N-(4-хлорофенил)-2-[1-(4-хлорофенил)-2,5-диоксопиролидин-3-илиден]-ацетамида на опухолевым клеткам (Т-клеточной лейкемии Jurkat, аденокарциномы молочной железы MCF-7, карциномы яичника Skov3, меланомы SK-Mel-28, немелкоклеточного рака легких SW-1573) in vitro. Синтезируемые соединения имеют различное цитотоксическое действие по отношению к используемых линий опухолевых клеток. Клетки лейкемии оказались наиболее чувствительными к этим производным. Цитотоксический эффект соединения 2 относительно Т-клеточной лейкемии Jurkat зависит від дозы соединения і продолжительности ее действия (3, 6, 24, 48 і 72 год). (3, 6, 24, 48 и 72 ч). Соединение 2 обладает цитотоксическим на лейкемические клетки линии Jurkat уже через 6 ч после ее добавления к культуре этих клеток (IC₅₀ = 66 мкМ), а через 24 часа ее цитотоксичность увеличивается (IC₅₀ = 40 мкМ). Выводы. Определено цитотоксического действия новых спиро-замещенных производных тиопирано[2,3-d]тиазола на опухолевые клетки in vitro. Соединение 2 обладает наивысшей активностью на лейкемические Т-клеточные линии Jurkat (IC₅₀ = 33,5 мкМ), в тоже время ее токсичность на псевдо-нормальных клетках млекопитающих была относительно невысокой. 2017 Article Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells / N.I. Zelisko, N.S. Finiuk, V.M. Shvets, Yu.O. Medvid, R.S. Stoika, R.B. Lesyk // Вiopolymers and Cell. — 2017. — Т. 33, № 4. — С. 282-290. — Бібліогр.: 25 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.00095A http://dspace.nbuv.gov.ua/handle/123456789/152984 547.818:547.489.4:542.91:615.359 en Вiopolymers and Cell Інститут молекулярної біології і генетики НАН України
institution	Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection	DSpace DC
language	English
topic	Bioorganic Chemistry Bioorganic Chemistry
spellingShingle	Bioorganic Chemistry Bioorganic Chemistry Zelisko, N.I. Finiuk, N.S. Shvets, V.M. Medvid, Yu.O. Stoika, R.S. Lesyk, R.B. Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells Вiopolymers and Cell
description	Aim. To evaluate the in vitro cytotoxicity of novel spiro-substituted thiopyrano[2,3-d]thiazoles towards tumor cells of different tissue origin. Methods. Organic synthesis; spectral methods; MTT test, statistical analysis. Results. In vitro screening of the cytotoxic activity of the 5’-carboxy-7’-aryl-1-aryl-3’,7’-dihydro-2H,2’H,5H-spiro[pyrolidin-3,6’-thiopyrano[2,3-d]thiazol]-2,2’,5-triones and N-(4-chlorophenyl)-2-[1-(4-chlorophenyl)-2,5-dioxopyrrolidin-3-ylidene]-acetamide was performed using various cancer cell lines (Jurkat human acute T-cell leukemia cell line, MCF-7 human breast adenocarcinoma cell line, Skov3 human ovarian carcinoma cells line, SK-Mel-28 human melanoma cells line, and SW-1573 human non-small-cell lung cancer cell line). The tested compounds possessed different cytotoxic action towards the studied tumor cells. Leukemia cells appeared to be more sensitive for the studied derivatives. The cytotoxic effect of the compound 2 towards Jurkat cells was shown to be dose- and time-dependent (3, 6, 24, 48 and 72 h). This compound demonstrated the cytotoxic action towards Jurkat cells as soon as in 6 h after its addition to the cultured cells (IC₅₀ = 66 μM), and its toxicity towards these cells was more prominent after 24 h treatment (IC₅₀= 40 μM). Conclusions. The panel of thiopyrano[2,3-d]thiazole derivatives was synthesized and screened for their cytotoxic activity in vitro towards tumor cells of different tissue origin. The compound 2 was found to be the most active agent with selectivity for the leukemia cells. This compound inhibits growth of the human acute T-cell leukemia cells of Jurkat line (IC₅₀ = 33.5 μM) and possesses relatively low toxicity towards the pseudo-normal mammalian cells.
format	Article
author	Zelisko, N.I. Finiuk, N.S. Shvets, V.M. Medvid, Yu.O. Stoika, R.S. Lesyk, R.B.
author_facet	Zelisko, N.I. Finiuk, N.S. Shvets, V.M. Medvid, Yu.O. Stoika, R.S. Lesyk, R.B.
author_sort	Zelisko, N.I.
title	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells
title_short	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells
title_full	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells
title_fullStr	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells
title_full_unstemmed	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells
title_sort	screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells
publisher	Інститут молекулярної біології і генетики НАН України
publishDate	2017
topic_facet	Bioorganic Chemistry
url	http://dspace.nbuv.gov.ua/handle/123456789/152984
citation_txt	Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells / N.I. Zelisko, N.S. Finiuk, V.M. Shvets, Yu.O. Medvid, R.S. Stoika, R.B. Lesyk // Вiopolymers and Cell. — 2017. — Т. 33, № 4. — С. 282-290. — Бібліогр.: 25 назв. — англ.
series	Вiopolymers and Cell
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fulltext	282 N. I. Zelisko, N. S. Finiuk, V. M. Shvets © 2017 N. I. Zelisko et al.; Published by the Institute of Molecular Biology and Genetics, NAS of Ukraine on behalf of Bio- polymers and Cell. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited UDC 547.818:547.489.4:542.91:615.359 Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells N. I. Zelisko1, N. S. Finiuk2, V. M. Shvets3, Yu. O. Medvid1, R. S. Stoika2, R. B. Lesyk1 1 Danylo Halytsky Lviv National Medical University 69, Pekarska Str., Lviv, Ukraine, 79010 2 Institute of Cell Biology, NAS of Ukraine 14/16, Drahomanov Str., Lviv, Ukraine, 79005 3 Zaporizhia State Medical University, Mayakovsky avenue 26, Zaporizhzhia, Ukraine, 69035 dr_r_lesyk@org.lviv.net, roman.lesyk@gmail.com Aim. To evaluate the in vitro cytotoxicity of novel spiro-substituted thiopyrano[2,3-d]thiazoles towards tumor cells of different tissue origin. Methods. Organic synthesis; spectral methods; MTT test, statistical analysis. Results. In vitro screening of the cytotoxic activity of the 5’-carboxy-7’-aryl-1-aryl-3’,7’-dihydro-2H,2’H,5H-spiro[pyrolidin-3,6’-thiopyrano[2,3-d] thiazol]-2,2’,5-triones and N-(4-chlorophenyl)-2-[1-(4-chlorophenyl)-2,5-dioxopyrrolidin-3- ylidene]-acetamide was performed using various cancer cell lines (Jurkat human acute T-cell leukemia cell line, MCF-7 human breast adenocarcinoma cell line, Skov3 human ovarian carcinoma cells line, SK-Mel-28 human melanoma cells line, and SW-1573 human non-small- cell lung cancer cell line). The tested compounds possessed different cytotoxic action towards the studied tumor cells. Leukemia cells appeared to be more sensitive for the studied deriva- tives. The cytotoxic effect of the compound 2 towards Jurkat cells was shown to be dose- and time-dependent (3, 6, 24, 48 and 72 h). This compound demonstrated the cytotoxic action towards Jurkat cells as soon as in 6 h after its addition to the cultured cells (IC50 = 66 μM), and its toxicity towards these cells was more prominent after 24 h treatment (IC50= 40 μM). Conclusions. The panel of thiopyrano[2,3-d]thiazole derivatives was synthesized and screened for their cytotoxic activity in vitro towards tumor cells of different tissue origin. The compound 2 was found to be the most active agent with selectivity for the leukemia cells. This compound inhibits growth of the human acute T-cell leukemia cells of Jurkat line (IC50 = 33.5 μM) and possesses relatively low toxicity towards the pseudo-normal mammalian cells. K e y w o r d s: thiopyrano[2,3-d]thiazoles, cytotoxic activity Bioorganic Chemistry ISSN 1993-6842 (on-line); ISSN 0233-7657 (print) Biopolymers and Cell. 2017. Vol. 33. N 4. P 282–290 doi: http://dx.doi.org/10.7124/bc.00095A mailto:dr_r_lesyk@org.lviv.net 283 Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells Introduction Chemotherapy is one of the most effective ways of treating cancer patients. Conventional anticancer drug discovery and development have been focused on the cytotoxic agents. The drug discovery paradigms selected agents that had significant cytostatic or cytotoxic activity towards tumor cell lines in vitro and caused tumor regression in vivo [1]. However, their application in cancer treatment is accompanied by frequent non-addressed actions lea ding to numerous negative side effects in the orga- nism [2]. Due to these effects, they de mon stra- te toxicity toward different normal cells of tissues and organs, including the bone marrow cells, mucous layer of the intestine, reproduc- tion glands, and hair follicles. 4-Thiazolidinones and related heterocycles have been demonstrated to be a perspective source of the innovative anticancer agents [3, 4]. In the previous studies, we identified a large group of substances with high anticancer potential and low toxicity [5, 6]. According to the obtained data [6], most of the studied sub- stances influenced cancer cell lines. 5-Ene-4- thiazolidinones were identified to possess the highest antitumor activity towards leukemia cells and induction of apoptosis in the mam- malian leukemia cells [7]. However, the mech- anisms of the anticancer activity of these com- pounds have not been studied yet. Previously it has been reported that PPARγ agonists, namely 4-thazolidinone derivatives, have a therapeutic potential in treatment of the inflammation and some cancers. This scien- tific assumption was successfully proved [8]. Among 4-thiazolidinones, there were also identified the effective inhibitors of the anti- apoptotic proteins Bcl-XL and BH3 interac- tions [9], the inhibitors of the interaction be- tween TNFα and the receptor TNFRc-1 [10], and the inhibitors of translation initiation [11]. A significant influence on the above mentioned pathogenic mechanisms of tumor growth de- termines perspectives of 4-thiazolidinones as potential anticancer drugs. The studied thiopyrano[2,3-d]thiazoles [12–14] keep the pharmacological profile of 5-ene-4-thiazolidinones and create new pos- sibilities for biological activities. Among the described thiopyrano[2,3-d]thiazole deriva- tives, there are anticancer [15–19], antitrypano- somal [20], antimycobacterial, antibacterial, and antifungal agents [21]. The main aim of this study was to measure the toxic effect in vitro of novel spiro-substi- tuted thiopyrano[2,3-d]thiazoles towards tumor cells of different tissue origin. Materials and Methods All chemicals used in the study were of the analytical grade and commercially available. All reagents and solvents were used without further purification and drying. Chemistry Series of rel-(5’R,6’R,7’R)-5’-carboxy-7’-aryl- 1 - a r y l - 3 ’ , 7 ’ - d i h y d r o - 2 H , 2 ’ H , 5 H - spiro[pyrolidin-3,6’-thiopyrano[2,3-d]thiazol]- 2,2’,5-triones (1a-e) for screening cytotoxi ci ty were synthesized via hetero-Diels-Alder reac- tion (Scheme 1) starting with trans-aconitic acid, corresponding aromatic amine and ap- propriate 5-arylidene-4-thioxo-2-thiazoli di no ne in the presence of the catalytic amount of hyd- roquinone (2–3 mg) for preventing polymeriza- tion processes, as it was described in our previ- 284 N. I. Zelisko, N. S. Finiuk, V. M. Shvets et al. ous work [22]. The 5-arylidene-4-thioxo-2-thi- azolidinones were synthesized via Knoevenagel condensation of 4-thioxo-2-thiazolidinone with the aromatic aldehydes [23]. Notably, we have identified 2-(2,5-dioxo- 1-arylpyrrolidin-3-ylidene)-N-aryl-acetamides as minor products of the mentioned one-pot three-component reaction. These compounds were easily removed during the recrystalliza- tion of crude products. We have found that the mentioned 2-(2,5-dioxo-1-arylpyrrolidin- 3-ylidene)-N-aryl-acetamides are formed with good yields via the interaction of trans-aconi- tic acid and appropriate aniline in the ratio 1:2. Thus, N-(4-chlorophenyl)-2-[1-(4-chloro phe- nyl)-2,5-dioxopyrrolidin-3-ylidene]-acetamide 2 was synthesized in the glacial acetic acid medium using 4-chloroaniline as amino com- ponent (Scheme 2). Screening of cytotoxic activity. Materials 10 mM stock solution of synthesized com- pounds was prepared in the dimethyl sulfoxide (DMSO, Sigma-Aldrich, USA), and additio- nal ly dissolved in culture medium prior to its addition to the cell culture. 3-(4,5-Dimethyl- thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) was 99.5 % pure. Cell culture medium DMEM was obtained from Sigma-Aldrich (USA), and RPMI – from APP (Austria). Fetal bovine serum (FBS) was obtained from APP (Austria), and Doxorubicin – from Pharma- che mie B.V. (the Netherlands). Cell cultures Human acute T-cell leukemia cells of Jurkat line, human breast adenocarcinoma cells of MCF-7 line, human ovarian carcinoma cells of Skov3 line, human melanoma cells of SK- S NH S O R1 COOH HOOC COOH NH2 R2 N S N H S O O O COOH R2 R1 + + 1 1a R1 = Br, R2 = Me 1b R1 = Br, R2 = F 1c R1 = Br, R2 = Cl 1d R1 = OMe, R2 = F 1e R1 =OMe, R2 = n-Bu AcOH hydroquinone Scheme.1. General synthetic path used for getting rel-(5’R,6’R,7’R)-5’-carboxy-7’-aryl-1-aryl-3’,7’-dihydro- 2H,2’H,5H-spiro[pyrolidin-3,6’-thiopyrano[2,3-d]thiazol]-2,2’,5-triones. COOH HOOC COOH NH2 Cl N O O O NH Cl Cl + 2 2 AcOH Scheme.2. Synthesis of N-(4-chlorophenyl)-2-[1-(4-chlorophenyl)-2,5-dioxopyrrolidin-3-ylidene]-acetamide. 285 Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells Mel-28 line, human non-small-cell lung cancer cells of SW-1573 line, transformed mouse fi- broblasts of L929 line, human embryonic kid- ney cells of HEK293 line were obtained from Cell culture collection at R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiology (Kyiv, Ukraine). Cells were grown in the RPMI or DME culture medium supplemented with 10 % fetal bovine serum. Cells were cultivated in the CO2-thermostate at 37 °C in atmosphere of 95 % air and 5 % CO2 at 100 % humidity. Cytotoxicity assay Cytotoxic activity of the synthesized com- pounds and doxorubicin used as a reference drug control towards cancer cell lines cul- tured in vitro was measured by the MTT test [24]. Tumor cells were seeded for 24 h in 96-well microtiter plates at a concentration of 2,000 of substrate-dependent cells/well or 10,000 of suspension cells/well in 100 μL. After that, cells were treated for 72 h with various additions of the synthesized com- pounds (0; 1; 10; 50; 100 μM). The MTT reagent, which is converted by mitochon- drial dehydrogenases to dark blue, water in- soluble MTT formazan, was used to measure the vitality of cells according to the manufac- turer’s protocol (Sigma-Aldrich, USA). The IC50 of tested compounds was calculated as a lethal concentration of drug decreasing cell vitality by 50 % in comparison with drug-free culture medium. Statistical analysis All data are presented as the mean ± standart deviation (SD). Results were analyzed and illustrated with GraphPad Prism (version 6; GraphPad Software, San Diego, CA). Statis ti- cal analyses were performed using two-way ANOVA with Bonferroni post-tests (tumor growth). P-value of <0.05 was considered as statistically significant. Results and Discussion Measuring of cytotoxic activity In vitro screening of cytotoxic activity of the synthesized compounds towards various tumor cell lines (human acute T-cell leukemia cells of Jurkat line, human breast adenocarcinoma cells of MCF-7 line, human ovarian carcinoma cells of Skov3 line, human melanoma cells of SK-Mel-28 lines, and human non-small-cell lung cancer cells of SW-1573 line) was per- formed using the MTT assay. The tested com- pounds were added to cultured cells in differ- ent final concentrations (0–100 μM), and then the cells were treated for 72 h (Fig. 1). The IC50 was calculated and used while comparing the values of the cytotoxic effect. The tested compounds were found to pos- sess different cytotoxic action towards human tumor cells of various tissue origin. The tumor cells of epithelial origin (MCF-7, Skov3, SK- Mel-28 and SW-1573 lines) were relatively resistant to the action of compounds 1a-e, 2 used in concentrations up to 100 μM (Fig. 1). The anticancer cytotoxic effect in vitro of the compounds 1a-e, 2 was also studied using human acute leukemia T-cells of Jurkat line. The compound 2 possessed cytotoxic activity towards Jurkat cells with the IC50 = 33.5 μM (Fig. 2). Thus, the cytotoxic activity of synthesized compounds 1a-e, 2 seems to depend on the targeted tumor cell line, although the leukemia 286 N. I. Zelisko, N. S. Finiuk, V. M. Shvets et al. cells appeared to be the most sensitive to the action of studied derivatives. Most of chemotherapeutic drugs demon- strate toxicity toward normal cells of tissues and organs, namely the bone marrow cells, cells of mucous layer of the intestine, repro- duction glands, and hair follicles [25]. We have measured the cytotoxic effect of the com- pounds under study towards non-tumor cells, such as transformed mouse fibroblasts of L929 line and pseudo-normal human embryonic kidney cells of HEK293 line (Fig. 3). There was no significant cytotoxic effect of used compounds towards HEK293 and L929 cells. Thus, one could suggest the lack of general Fig. 2. Results of testing cytotoxicity of compounds to- wards human acute T-cell leukemia cells of Jurkat line. After a total time of experiment (72 h) cytotoxic effect was measured by the MTT assay. Fig. 1. Results of testing cytotoxicity of compounds towards different tumor cell lines. After a total time of experiment (72 h), cytotoxic effect was measured by the MTT assay. 287 Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells toxicity of novel synthetic compounds towards normal cells of tissues and organs. We found that the cytotoxic effect of com- pound 2 towards human acute T-cell leukemia cells of Jurkat line was dose- and time-depen- dent (3, 6, 24, 48 and 72 h). Compound 2 showed such effect as soon as in 6 h after starting cell treatment (IC50 = 66 μM), where- as its toxicity towards Jurkat cells was more prominent after 24 h treatment (IC50 = 40 μM) (Fig. 4). Conclusion Novel spiro-substituted thiopyrano[2,3-d]thia- zole derivatives were synthesized and tested for their cytotoxic activity towards human tumor cells in vitro. Generally, they possessed low toxicity towards tumor cells. Compound 2 was found to be the most cytotoxic agent specifically towards the human acute T-cell leukemia cells of Jurkat line (IC50 = 33.5 μM). A low toxicity of this compound towards the pseudo-normal human embryonic kidney cells of HEK293 line should be noted. Acknowledgement The publication contains the results of studies conducted by President’s of Ukraine grant for competitive projects F-63 of the State Fund for Fundamental Research. The investigation was also supported by Cedars Sinai Medical Center’s International Research and Innovation in Medicine Program, the Association for Regional Cooperation in the Fields of Health, Science and Technology (RECOOP HST Association) and the participating Cedars – Sinai Medical Center – RECOOP Research Centers (CRRC). J u r k a t C o n c e n t r a t i o n , m M -f o ld o f g ro w th 0 20 40 60 80 100 0.0 0.2 0.4 0.6 0.8 1.0 1.2 7 2 h 4 8 h 2 4 h 6 h 3 h Fig. 4. Exposure time-dependency of compound 2 cyto- toxicity. Pulsing experiments were performed using hu- man acute T-cell leukemia cells of Jurkat line treated for 3, 6, 24, 48 and 72 h with compound 2. Cytotoxic action was determined using MTT assay. Fig. 3. Results of testing cytotoxicity of compounds towards non-tumor mammalian cells. 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Eur Food Res Technol. 2009;229:247–253. 25. Havrylyuk D, Zimenkovsky B, Vasylenko O, Gzel- la A, Lesyk R. Synthesis of new 4-thiazolidinone, pyrazoline, and isatin-based conjugates with prom- ising antitumor activity. J Med Chem. 2012;55:8630– 8641. Скринінг спіро-заміщених тіопірано[2,3-d] тіазолів щодо їх цитотоксичної дії на пухлинні клітини Н. І. Зеліско, Н. С. Фінюк, В. М. Швець, Ю. О. Медвідь, Р. С. Стойка, Р. Б. Лесик Мета. Вивчення цитотоксичної активності нових спі- ро-заміщених похідних тіопірано[2,3-d]тіазолу на пух- линні клітин різного походження. Методи. Органічний синтез; спектральні методи; МТТ тест, статистичний аналіз. Результати. Проведено дослідження іn vitro 5’-карбокси-7’-арил-1-арил-3’,7’-дигідро-2H,2’H,5H- спіро[піролідин-3,6’-тіопірано[2,3-d]тіазол]-2,2’,5- тріонів і N-(4-хлорофеніл)-2-[1-(4-хлорофеніл)-2,5- діоксопіролідин-3-іліден]-ацетаміду щодо різних ліній пухлинних клітин (Т-клітинної лейкемії Jurkat, адено- карциноми молочної залози MCF-7, карциноми яєч- ника Skov3, меланоми SK-Mel-28, недрібноклітинного раку легені SW-1573). Синтезовані сполуки мають різну цитотоксичну дію щодо використаних ліній пухлинних клітин. Клітини лейкемії виявилися най- більш чутливими до дії цих похідних. Цитотоксичний ефект сполуки 2 щодо Т-клітинної лейкемії Jurkat за- лежить від дози сполуки і тривалості її дії (3, 6, 24, 48 і 72 год). Сполука 2 володіє цитотоксичною дією на лейкемічні клітини лінії Jurkat вже через 6 год після її додавання до культури цих клітин (IC50 = 66 μМ), а через 24 год її цитотоксичність зростає (IC50 = 40 μМ). Висновки. Проведено скринінг нових спіро-заміщених похідних тіопірано[2,3-d]тіазолу щодо їх цитотоксич- ної дії на пухлинні клітини іn vitro. Сполука 2 володіє найвищою активністю у лейкемічних Т-клітинах лінії Jurkat (IC50 = 33,5 мкМ), тоді як її токсичність щодо псевдо-нормальних клітин ссавців є відносно низькою. К л юч ов і с л ов а: тіопірано[2,3-d]тіазоли, проти- пухлинна активність Скрининг спиро-замещенных тиопирано[2,3-d] тиазолов относительно их цитотоксического действия на опухолевые клетки Н. И. Зелиско, Н. С. Финюк, В. Н. Швець, Ю. О. Медвидь, Р. С. Стойка, Р. Б. Лесык Цель. Изучение цитотоксической активности новых спиро-замещенных производных тиопирано[2,3-d] тиазола на опухолевые клетки различной этиологии. Методы. Органический синтез; спектральные методы; МТТ тест, статистический анализ. Результаты. Исследовано действие 5’-карбокси-7’-арил-1-арил- 3’,7’-дигидро-2H,2’H,5H-спиро[пиролидин-3,6’- тиопирано[2,3-d]тиазол]-2,2’,5-трионов и N-(4- хлорофенил ) - 2 - [ 1 - ( 4 - хлорофенил ) - 2 , 5 - диоксопиролидин-3-илиден]-ацетамида на опухолевым клеткам (Т-клеточной лейкемии Jurkat, аденокарциномы 290 N. I. Zelisko, N. S. Finiuk, V. M. Shvets et al. молочной железы MCF-7, карциномы яичника Skov3, меланомы SK-Mel-28, немелкоклеточного рака легких SW-1573) in vitro. Синтезируемые соединения имеют различное цитотоксическое действие по отношению к используемых линий опухолевых клеток. Клетки лейкемии оказались наиболее чувствительными к этим производным. Цитотоксический эффект соединения 2 относительно Т-клеточной лейкемии Jurkat зависит від дозы соединения і продолжительности ее действия (3, 6, 24, 48 і 72 год). (3, 6, 24, 48 и 72 ч). Соединение 2 обладает цитотоксическим на лейкемические клетки линии Jurkat уже через 6 ч после ее добавления к культуре этих клеток (IC50 = 66 мкМ), а через 24 часа ее цитотоксичность увеличивается (IC50 = 40 мкМ). Выводы. Определено цитотоксического действия но- вых спиро-замещенных производных тиопирано[2,3-d] тиазола на опухолевые клетки in vitro. Соединение 2 обладает наивысшей активностью на лейкемические Т-клеточные линии Jurkat (IC50 = 33,5 мкМ), в тоже время ее токсичность на псевдо-нормальных клетках млекопитающих была относительно невысокой. К л юч е в ы е с л ов а: тиопирано[2,3-d]тиазолы, про- тивоопухолевая активность. Received 24.04.2017

Screening of spiro-substituted thiopyrano[2,3-d]thiazoles for their cytotoxic action on tumor cells

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