zldmЗаводская лаборатория. Диагностика материаловIndustrial laboratory. Diagnostics of materials1028-68612588-0187ООО «Издательство «ТЕСТ-ЗЛ»10.26896/1028-6861-2023-89-12-5-12zldm-2076Research ArticleАНАЛИЗ ВЕЩЕСТВАSUBSTANCES ANALYSISХроматографическое определение монойодацетата в липидных наноструктурахChromatographic determination of iodoacetate in lipid nanostructuresКоршуновД. А.KorshunovD. A.

Дмитрий Афанасьевич Коршунов

634009, Томск, Кооперативный пер., д. 5

Dmitry A. Korshunov

5, Cooperativny per., Tomsk, 634009

ieved@ya.ruКондаковаИ. В.KondakovaI. A.

Ирина Викторовна Кондакова

634009, Томск, Кооперативный пер., д. 5

Irina V. Kondakova

5, Cooperativny per., Tomsk, 634009

СиденкоЕ. А.SidenkoE. A.

Евгения Александровна Сиденко

634009, Томск, Кооперативный пер., д. 5

Evgenia A. Sidenko

5, Cooperativny per., Tomsk, 634009

СередаЕ. Е.SeredaE. E.

Елена Евгеньевна Середа

634009, Томск, Кооперативный пер., д. 5; 634050, Томск, Московский тракт, д. 2.

Elena E. Sereda

5, Cooperativny per., Tomsk, 634009; 2, Moskovsky trakt, Tomsk, 634050

ЗолотухинаН. Ю.ZolotukhinaN. Yu.

Наталья Юрьевна Золотухина

634050, Томск, просп. Ленина, д. 30

Natalia Yu. Zolotukhina

30, prosp. Lenina, Tomsk, 634050

Научно-исследовательский институт онкологии Томский национальный исследовательский медицинский центр РАНРоссияCancer Research Institute, Tomsk National Research Medical CenterRussian FederationНаучно-исследовательский институт онкологии Томский национальный исследовательский медицинский центр РАН; Сибирский государственный медицинский университетРоссияCancer Research Institute, Tomsk National Research Medical Center; Siberian State Medical UniversityRussian FederationНациональный исследовательский Томский политехнический университетРоссияNational Research Tomsk Polytechnic UniversityRussian Federation2023181220238912512Copyright © Коршунов Д.А., Кондакова И.В., Сиденко Е.А., Середа Е.Е., Золотухина Н.Ю., 20232023Коршунов Д.А., Кондакова И.В., Сиденко Е.А., Середа Е.Е., Золотухина Н.Ю.Korshunov D.A., Kondakova I.A., Sidenko E.A., Sereda E.E., Zolotukhina N.Y.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.zldm.ru/jour/article/view/2076

Для уменьшения системной токсичности и улучшения фармакокинетики противоопухолевых препаратов часто используют их липосомальную форму. Монойодацетат натрия (МЙА) — ингибитор гликолиза — противоопухолевый препарат, находящийся на стадии клинических испытаний. МЙА инкапсулировали в липосомы (липосомальная эмульсия содержала 1,2-дипальмитоил-глицеро-3-фосфохолин (ДПФХ) и холестерин (Хол) в молярном соотношении 9:0,8) методом экструзии: в зависимости от условий диаметр полученных липосом составил 100 или 400 нм. Полученные липосомы очищали от невключенного МЙА путем диализа в течение 3 ч. Для определения МЙА, содержащегося непосредственно в липосомах, необходимо сначала отделить липосомы от межлипосомальной жидкости, которая также содержит невключенный МЙА. Для этой цели использовали сорбцию липосом на активированном угле. Затем необходимо разрушить оболочку липосом: из исследованных агентов (ПАВ, бензалкония хлорид, апротонные растворители) наиболее эффективным для этой цели оказался диоксан. Разработана методика определения МЙА в липосомальной форме в диапазоне 31,3 мкг/мл – 7,9 мг/мл с пределом обнаружения 5,6 мкг/мл, обеспечивающая разделение МЙА, холестерина и фосфолипидов, с использованием метода хроматографии гидрофильного взаимодействия со спектрофотометрическим детектированием на длине волны λ = 260 нм. В качестве подвижной фазы использовали раствор ацетонитрила в воде в объемном соотношении 50:50 с добавлением модификаторов — уксусной кислоты (60 ммоль/л) и формиата аммония (30 ммоль/л), неподвижная фаза — аминопропилсиликагель. Установлено, что независимо от диаметра доза МЙА в 1 мл липосом составила 0,20 – 0,23 мг. Для использования в дальнейших исследованиях выбраны липосомы диаметром 400 нм, более мягкие условия получения которых обеспечивают меньшие степень окисления липидов и потери МЙА.

A liposomal form of anticancer drugs is often used to improve pharmacokinetics and reduce systemic toxicity of the drugs. The goal of the study is to develop a method for quantitative analysis of a liposomal form of sodium iodoacetate (IA), glycolysis inhibitor, which exhibits a pronounced antitumor activity. Liposomes were prepared by extrusion at a temperature of 25 – 55°C under argon pressure ranged from 2 to 10 MPa. The obtained liposomes were purified from the non-incorporated component using dialysis. The method of HPLC was used to analyze the inhibitor solution in liposomes. The method of hydrophilic interaction chromatography revealed a high selectivity of iodoacetate with aminopropyl silica gel as a stationary phase. The best option for analysis was to use a spectrophotometric detector. The results of analysis showed that the dose of the inhibitor in 1 ml of liposomes was 0.20 – 0.23 mg regardless of the liposome size. In terms of the weight of an animal, the amount of iodoacetate was 8 – 9 mg/kg. The analysis of liposomes by the developed method showed that the highest yield and a high degree of purification is attained at low temperature (no more than 40°C) and duration of dialysis for about 3 h. For these purposes, the use of liposomes with a diameter of 400 nm turned out to be the best option.

ингибиторы гликолизамонойодацетатопределениелипосомыВЭЖХglycolysis inhibitorssodium iodoacetatedeterminationliposomesHPLCReferencesSun D., Gao W., Hu H., Zhou S. Why 90% of clinical drug development fails and how to improve it? / Acta. Pharm. Sin. B. 2022. V. 12. N 7. P. 3049 – 3062. DOI: 10.1016/j.apsb.2022.02.002Sun D., Gao W., Hu H., Zhou S. Why 90% of clinical drug development fails and how to improve it? / Acta. Pharm. Sin. B. 2022. V. 12. N 7. P. 3049 – 3062. DOI: 10.1016/j.apsb.2022.02.002Veselov V. V., Nosyrev A. E., Jicsinszky L., et al. Targeted Delivery Methods for Anticancer Drugs / Cancers. 2022. V. 14. N 3. 622. DOI: 10.3390/cancers14030622Veselov V. V., Nosyrev A. E., Jicsinszky L., et al. Targeted Delivery Methods for Anticancer Drugs / Cancers. 2022. V. 14. N 3. 622. DOI: 10.3390/cancers14030622Tran S., DeGiovanni P. J., Piel B., Rai P. Cancer nanomedicine: a review of recent success in drug delivery / Clin. Transl. Med. 2017. V. 6. N 1. 44. DOI: 10.1186/s40169-017-0175-0Tran S., DeGiovanni P. J., Piel B., Rai P. Cancer nanomedicine: a review of recent success in drug delivery / Clin. Transl. Med. 2017. V. 6. N 1. 44. DOI: 10.1186/s40169-017-0175-0Liu Y., Li J., Chen M., et al. Palladium-based nanomaterials for cancer imaging and therapy / Theranostics. 2020. V. 10. N 22. P. 10057 – 10074. DOI: 10.7150/thno.45990Liu Y., Li J., Chen M., et al. Palladium-based nanomaterials for cancer imaging and therapy / Theranostics. 2020. V. 10. N 22. P. 10057 – 10074. DOI: 10.7150/thno.45990Cui G., Wu J., Lin J., et al. Graphene-based nanomaterials for breast cancer treatment: promising therapeutic strategies / J. Nanobiotechnol. 2021 V. 19. N 1. 211. DOI: 10.1186/s12951-021-00902-8Cui G., Wu J., Lin J., et al. Graphene-based nanomaterials for breast cancer treatment: promising therapeutic strategies / J. Nanobiotechnol. 2021 V. 19. N 1. 211. DOI: 10.1186/s12951-021-00902-8Zhu L., Zhao J., Guo Z., et al. Applications of Aptamer-Bound Nanomaterials in Cancer Therapy / Biosensors. 2021. V. 11. N 9. 344. DOI: 10.3390/bios11090344Zhu L., Zhao J., Guo Z., et al. Applications of Aptamer-Bound Nanomaterials in Cancer Therapy / Biosensors. 2021. V. 11. N 9. 344. DOI: 10.3390/bios11090344Luo L., Wang H., Tian W., et al. Targeting ferroptosis-based cancer therapy using nanomaterials: strategies and applications / Theranostics. 2021. V. 11. N 20. P. 9937 – 9952. DOI: 10.7150/thno.65480Luo L., Wang H., Tian W., et al. Targeting ferroptosis-based cancer therapy using nanomaterials: strategies and applications / Theranostics. 2021. V. 11. N 20. P. 9937 – 9952. DOI: 10.7150/thno.65480Bor G., Mat Azmi I. D., Yaghmur A. Nanomedicines for cancer therapy: current status, challenges and future prospects / Ther Delivery. 2019. V. 10. N 2. P. 113 – 132. DOI: 10.4155/tde-2018-0062Bor G., Mat Azmi I. D., Yaghmur A. Nanomedicines for cancer therapy: current status, challenges and future prospects / Ther Delivery. 2019. V. 10. N 2. P. 113 – 132. DOI: 10.4155/tde-2018-0062Korshunov D. A., Kondakova I. V., Shashova E. E. Modern perspective on metabolic reprogramming in malignant neoplasms / Biochemistry (Moscow). 2019. V. 84. N 10. P. 1129 – 1142. DOI: 10.1134/S000629791910002XKorshunov D. A., Kondakova I. V., Shashova E. E. Modern perspective on metabolic reprogramming in malignant neoplasms / Biochemistry (Moscow). 2019. V. 84. N 10. P. 1129 – 1142. DOI: 10.1134/S000629791910002XMikubo M., Inoue Y., Liu G., Tsao M. S. Mechanism of Drug Tolerant Persister Cancer Cells: The Landscape and Clinical Implication for Therapy / J. Thorac. Oncol. 2021. V. 16. N 11. P. 1798 – 1809. DOI: 10.1016/j.jtho.2021.07.017Mikubo M., Inoue Y., Liu G., Tsao M. S. Mechanism of Drug Tolerant Persister Cancer Cells: The Landscape and Clinical Implication for Therapy / J. Thorac. Oncol. 2021. V. 16. N 11. P. 1798 – 1809. DOI: 10.1016/j.jtho.2021.07.017Faubert B., Solmonson A., De Berardinis R. J. Metabolic reprogramming and cancer progression / Science. 2020. V. 368. N 6487. eaaw5473. DOI: 10.1126/science.aaw5473Faubert B., Solmonson A., De Berardinis R. J. Metabolic reprogramming and cancer progression / Science. 2020. V. 368. N 6487. eaaw5473. DOI: 10.1126/science.aaw5473Cheng Z., Li M., Dey R., Chen Y. Nanomaterials for cancer therapy: current progress and perspectives / J. Hematol. Oncol. 2021. V. 14. N 1. 85. DOI: 10.1186/s13045-021-01096-0Cheng Z., Li M., Dey R., Chen Y. Nanomaterials for cancer therapy: current progress and perspectives / J. Hematol. Oncol. 2021. V. 14. N 1. 85. DOI: 10.1186/s13045-021-01096-0Sun X., Peng Y., Zhao J., et al. Discovery and development of tumor glycolysis rate-limiting enzyme inhibitors / Bioorg. Chem. 2021. V. 112. 104891. DOI: 10.1016/j.bioorg.2021.104891Sun X., Peng Y., Zhao J., et al. Discovery and development of tumor glycolysis rate-limiting enzyme inhibitors / Bioorg. Chem. 2021. V. 112. 104891. DOI: 10.1016/j.bioorg.2021.104891Korshunov D. A., Kondakova I. V., Klimov I. A., Ivanov V. V. Glycolysis inhibitors monoiodoacetate and 2-deoxyglucose as antitumor agents: experimental study on lewis lung carcinoma model / Bull. Exp. Biol. Med. 2018. V. 165. N 5. P. 695 – 697. DOI: 10.1007/s10517-018-4244-1Korshunov D. A., Kondakova I. V., Klimov I. A., Ivanov V. V. Glycolysis inhibitors monoiodoacetate and 2-deoxyglucose as antitumor agents: experimental study on lewis lung carcinoma model / Bull. Exp. Biol. Med. 2018. V. 165. N 5. P. 695 – 697. DOI: 10.1007/s10517-018-4244-1Brennan S., Esposito S., Abdelaziz M. I. M., et al. Selective protein kinase C inhibition switches time-dependent glucose cardiotoxicity to cardioprotection / Front. Cardiovasc. Med. 2022. V. 9. 997013. DOI: 10.3389/fcvm.2022.997013Brennan S., Esposito S., Abdelaziz M. I. M., et al. Selective protein kinase C inhibition switches time-dependent glucose cardiotoxicity to cardioprotection / Front. Cardiovasc. Med. 2022. V. 9. 997013. DOI: 10.3389/fcvm.2022.997013Yashin Ya., Vedenin A., Yashin A. HPLC and ultra-HPLC: a state and prospects / Analitika. 2015. N 2(21). P. 70 – 84 [in Russian].Yashin Ya., Vedenin A., Yashin A. HPLC and ultra-HPLC: a state and prospects / Analitika. 2015. N 2(21). P. 70 – 84 [in Russian].Lau B. P., Becalski A. Determination of iodoacetic acid using liquid chromatography/electrospray tandem mass spectrometry / Rapid. Commun. Mass. Spectrom. 2008. V. 22. N 12. P. 1787 —1791. DOI: 10.1002/rcm.3547Lau B. P., Becalski A. Determination of iodoacetic acid using liquid chromatography/electrospray tandem mass spectrometry / Rapid. Commun. Mass. Spectrom. 2008. V. 22. N 12. P. 1787 —1791. DOI: 10.1002/rcm.3547Guo Y. A survey of polar stationary phases for hydrophilic interaction chromatography and recent progress in understanding retention and selectivity / Biomed. Chromatogr. 2022. V. 36. N 4. e5332. DOI: 10.1002/bmc.5332Guo Y. A survey of polar stationary phases for hydrophilic interaction chromatography and recent progress in understanding retention and selectivity / Biomed. Chromatogr. 2022. V. 36. N 4. e5332. DOI: 10.1002/bmc.5332Singh D., Pahwa S. A review on physico-chemical parameters of liposomal doxorubicin / Int. J. Appl. Pharm. 2020. V. 12. N 2. P. 1 – 5. DOI: 10.22159/ijap.2020v12i2.35330Singh D., Pahwa S. A review on physico-chemical parameters of liposomal doxorubicin / Int. J. Appl. Pharm. 2020. V. 12. N 2. P. 1 – 5. DOI: 10.22159/ijap.2020v12i2.35330Iodoacetic acid. CAMEO Chemicals database, 2023. https:// cameochemicals.noaa.gov/chemical/20524 (accessed March 1, 2023).Iodoacetic acid. CAMEO Chemicals database, 2023. https:// cameochemicals.noaa.gov/chemical/20524 (accessed March 1, 2023).Milto I. V., Sukhodolo I. V., Usov V. Y. Mononuclear phagocytes of rat liver and lung after intravenous introduction of suspension of magnetite nanoparticles / Cell Tissue Biol. 2012. V. 6. N 5 – 6. P. 490 – 497. DOI: 10.1134/S1990519X12050082Milto I. V., Sukhodolo I. V., Usov V. Y. Mononuclear phagocytes of rat liver and lung after intravenous introduction of suspension of magnetite nanoparticles / Cell Tissue Biol. 2012. V. 6. N 5 – 6. P. 490 – 497. DOI: 10.1134/S1990519X12050082Technical Guide for the Elaboration of Monographs. 8th Ed. — European Pharmacopoeia, 2022. P. 45 – 56.Technical Guide for the Elaboration of Monographs. 8th Ed. — European Pharmacopoeia, 2022. P. 45 – 56.United States Pharmacopeia. General Chapter, •621• Chromatography. USP-NF. — Rockville, MD: United States Pharmacopeia. 2023. DOI: 10.31003/USPNF_M99380_06_01United States Pharmacopeia. General Chapter, •621• Chromatography. USP-NF. — Rockville, MD: United States Pharmacopeia. 2023. DOI: 10.31003/USPNF_M99380_06_01Chromatography / Russian Federation State Pharmacopoeia, XIV ed. V. 1. P. 845 – 872 [in Russian].Chromatography / Russian Federation State Pharmacopoeia, XIV ed. V. 1. P. 845 – 872 [in Russian].Iodoacetic acid (64-69-7). NIST Chemistry WebBook. NIST Standard Reference Database No. 69, Feb 2015 Release. — Washington, DC: US Sec. Commerce, 2018. https://webbook.nist.gov/cgi/cbook.cgi?ID=C64697&Mask=400#UV-Vis-Spec (accessed March 1, 2023).Iodoacetic acid (64-69-7). NIST Chemistry WebBook. NIST Standard Reference Database No. 69, Feb 2015 Release. — Washington, DC: US Sec. Commerce, 2018. https://webbook.nist.gov/cgi/cbook.cgi?ID=C64697&Mask=400#UV-Vis-Spec (accessed March 1, 2023).Paternostre M. T., Roux M., Rigaud J. L. Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 1. Solubilization of large unilamellar liposomes (prepared by reverse-phase evaporation) by triton X-100, octyl glucoside, and sodium cholate / Biochemistry. 1988. V. 27. N 8. P. 2668 – 2677. DOI: 10.1021/bi00408a006Paternostre M. T., Roux M., Rigaud J. L. Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents. 1. Solubilization of large unilamellar liposomes (prepared by reverse-phase evaporation) by triton X-100, octyl glucoside, and sodium cholate / Biochemistry. 1988. V. 27. N 8. P. 2668 – 2677. DOI: 10.1021/bi00408a006Bjørnestad V. A., Lund R. Pathways of Membrane Solubilization: A Structural Study of Model Lipid Vesicles Exposed to Classical Detergents / Langmuir. 2023. V. 39. N 11. P. 3914 – 3933. DOI: 10.1021/acs.langmuir.2c03207Bjørnestad V. A., Lund R. Pathways of Membrane Solubilization: A Structural Study of Model Lipid Vesicles Exposed to Classical Detergents / Langmuir. 2023. V. 39. N 11. P. 3914 – 3933. DOI: 10.1021/acs.langmuir.2c03207Bjørnestad V. A., Soto-Bustamante F., Tria G., et al. Beyond the standard model of solubilization: Non-ionic surfactants induce collapse of lipid vesicles into rippled bilamellar nanodiscs / J. Colloid Interface Sci. 2023. V. 641. P. 553 – 567. DOI: 10.1016/j.jcis.2023.03.037Bjørnestad V. A., Soto-Bustamante F., Tria G., et al. Beyond the standard model of solubilization: Non-ionic surfactants induce collapse of lipid vesicles into rippled bilamellar nanodiscs / J. Colloid Interface Sci. 2023. V. 641. P. 553 – 567. DOI: 10.1016/j.jcis.2023.03.037Labeta M. O., Fernandez N., Festenstein H. Solubilisation effect of Nonidet P-40, triton X-100 and CHAPS in the detection of MHC-like glycoproteins / J. Immunol. Methods. 1988. V. 112. N 1. P. 133 – 138. DOI: 10.1016/0022-1759(88)90043-9Labeta M. O., Fernandez N., Festenstein H. Solubilisation effect of Nonidet P-40, triton X-100 and CHAPS in the detection of MHC-like glycoproteins / J. Immunol. Methods. 1988. V. 112. N 1. P. 133 – 138. DOI: 10.1016/0022-1759(88)90043-9Kim M., Hatt J. K., Weigand M. R., et al. Genomic and Transcriptomic Insights into How Bacteria Withstand High Concentrations of Benzalkonium Chloride Biocides / Appl. Environ. Microbiol. 2018. V. 84. N 12. e00197 – 18. DOI: 10.1128/AEM.00197-18Kim M., Hatt J. K., Weigand M. R., et al. Genomic and Transcriptomic Insights into How Bacteria Withstand High Concentrations of Benzalkonium Chloride Biocides / Appl. Environ. Microbiol. 2018. V. 84. N 12. e00197 – 18. DOI: 10.1128/AEM.00197-18Wessels S., Ingmer H. Modes of action of three disinfectant active substances: a review / Regul. Toxicol. Pharmacol. 2013. V. 7. N 3. P. 56 – 67. DOI: 10.1016/j.yrtph.2013.09.006Wessels S., Ingmer H. Modes of action of three disinfectant active substances: a review / Regul. Toxicol. Pharmacol. 2013. V. 7. N 3. P. 56 – 67. DOI: 10.1016/j.yrtph.2013.09.006

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