inovmedИнновационная медицина КубаниInnovative Medicine of Kuban2541-9897Scientific Research Institute – Ochapovsky Regional Clinical Hospital No. 110.35401/2541-9897-2026-11-1-148-156inovmed-1455Research ArticleОБЗОРЫREVIEWSОсобенности дизайна доклинических исследований препаратов генной терапии in vivo. Часть 2: фармакокинетические и токсикологические исследованияDesign Features of Nonclinical Studies for In Vivo Gene Therapy Medicinal Products. Part 2: Pharmacokinetic and Toxicological Studieshttps://orcid.org/0000-0001-8377-9205РачинскаяО. А.RachinskayaO. A.

Рачинская Ольга Анатольевна - к. б. н., ведущий эксперт лаборатории биомедицинских клеточных продуктов.

127051, Москва, Петровский б-р, д. 8, стр. 2

Olga A. Rachinskaya - Cand. Sci. (Biol.), Leading Expert, Laboratory of Biomedical Cell Products, Scientific Centre for Expert Evaluation of Medicinal Products.

Petrovskii bulvar 8/2, Moscow, 127051

Rachinskaya@expmed.ruhttps://orcid.org/0000-0002-9585-3545МельниковаЕ. В.MelnikovaE. V.

Мельникова Екатерина Валерьевна - к. б. н., начальник лаборатории биомедицинских клеточных продуктов.

Москва

Ekaterina V. Melnikova - Cand. Sci. (Biol.), Head of the Laboratory of Biomedical Cell Products, Scientific Centre for Expert Evaluation of Medicinal Products.

Moscow

https://orcid.org/0000-0003-4891-973XМеркуловВ. А.MerkulovV. A.

Меркулов Вадим Анатольевич - д. м. н., профессор, заместитель генерального директора.

Москва

Vadim A. Merkulov - Dr. Sci. (Med.), Professor, Deputy General Director, Scientific Centre for Expert Evaluation of Medicinal Products.

Moscow

Научный центр экспертизы средств медицинского примененияРоссияScientific Centre for Expert Evaluation of Medicinal ProductsRussian Federation202630032026111148156Copyright © Рачинская О.А., Мельникова Е.В., Меркулов В.А., 20262026Рачинская О.А., Мельникова Е.В., Меркулов В.А.Rachinskaya O.A., Melnikova E.V., Merkulov V.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.innovmedkub.ru/jour/article/view/1455

Статья посвящена особенностям дизайна доклинических исследований генотерапевтических лекарственных препаратов in vivo, основанных на введении векторов с рекомбинантными нуклеиновыми кислотами в организм пациента для коррекции генетических нарушений и терапии онкологических заболеваний. В настоящей работе отмечаются основные аспекты проведения фармакокинетических и токсикологических исследований, выявленные в результате анализа экспертных отчётов мировых производителей генотерапевтических лекарственных препаратов и затрагивающие такие вопросы, как выбор дозы и способа введения препарата; выявление специфических исследований, характерных для всех in vivo генотерапевтических лекарственных препаратов и обоснование перечня исследований для препаратов с разными видами векторов; продолжительность исследований и выбор адекватных моделей для демонстрации безопасности продукта и выявления возможных нежелательных явлений от его применения.

The article focuses on the specific design features of nonclinical studies (NCS) of in vivo gene therapy medicinal products which involve the administration of vectors with recombinant nucleic acids into the patient’s body for the correction of genetic disorders and treatment of oncological diseases. This work highlights the key aspects of conducting pharmacokinetic and toxicological studies, identified through the analysis of expert reports from global gene therapy medicinal products manufacturers. These aspects include dose selection and route of administration; identification of specific studies applicable to all in vivo gene therapy medicinal products and the justification of study lists for products using different types of vectors; as well as duration of studies and the selection of appropriate models to demonstrate product safety and to detect potential adverse effects associated with their use.

генотерапевтические лекарственные препаратыдоклинические исследованияфармакокинетические исследованиятоксикологические исследованиявектортрансгенgene therapy productsnonclinical studiespharmacokinetic studiestoxicological studiesvectortransgeneРабота выполнена в рамках государственного задания ФГБУ «НЦЭСМП» Минздрава России № 056-00001-25-01 на проведение прикладных научных исследований (номер государственного учёта НИР 124022200093-9).The study was carried out within the framework of FSBI “SCEEMP” No. № 056-00001-25-01 for applied scientific research (Research State Registration Number No. 124022200093-9).ReferencesEuropean Medicines Agency. Guideline on the quality, non-clinical and clinical aspects of gene therapy medicinal products. European Medicines Agency; 2018. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-quality-non-clinical-and-clinical-aspects-gene-therapy-medicinal-products_en.pdfEuropean Medicines Agency. Guideline on the quality, non-clinical and clinical aspects of gene therapy medicinal products. European Medicines Agency; 2018. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/scientific-guideline/guideline-quality-non-clinical-and-clinical-aspects-gene-therapy-medicinal-products_en.pdfРачинская О.А., Мельникова Е.В., Меркулов В.А. Особенности дизайна доклинических исследований препаратов генной терапии in vivo. Часть 1: фармакологические исследования. Инновационная медицина Кубани. 2025;10(4):113-120. https://doi.org/10.35401/2541-9897-2025-10-4-113-120Rachinskaya O.A., Melnikova E.V., Merkulov V.A. Design Features of Nonclinical Studies of In Vivo Gene Therapy Medicinal Products. Part 1: Pharmacological studies. Innovative Medicine of Kuban. 2025;10(4):113-120. https://doi.org/10.35401/2541-9897-2025-10-4-113-120International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. ICH Harmonised Guideline: Nonclinical Biodistribution Considerations for Gene Therapy Products S12. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; 2023. Accessed April 13, 2025. https://database.ich.org/sites/default/files/ICH_S12_Step4_Guideline_2023_0314.pdfInternational Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. ICH Harmonised Guideline: Nonclinical Biodistribution Considerations for Gene Therapy Products S12. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; 2023. Accessed April 13, 2025. https://database.ich.org/sites/default/files/ICH_S12_Step4_Guideline_2023_0314.pdfMoffit JS, Blanset DL, Lynch JL, et al. Regulatory consideration for the nonclinical safety assessment of gene therapies. Hum Gene Ther. 2022;33(21–22):1126–1141. PMID: 35994386. PMCID: PMC9700330. https://doi.org/10.1089/hum.2022.090Moffit JS, Blanset DL, Lynch JL, et al. Regulatory consideration for the nonclinical safety assessment of gene therapies. Hum Gene Ther. 2022;33(21–22):1126–1141. PMID: 35994386. PMCID: PMC9700330. https://doi.org/10.1089/hum.2022.090European Medicines Agency. ICH Considerations: General Principles to Address Virus and Vector Shedding. European Medicines Agency; 2009. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/scientific-guideline/international-conference-harmonisation-technical-requirements-registration-pharmaceuticals-human-use-considerations-general-principles-address-virus-and-vector-shedding_en.pdfEuropean Medicines Agency. ICH Considerations: General Principles to Address Virus and Vector Shedding. European Medicines Agency; 2009. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/scientific-guideline/international-conference-harmonisation-technical-requirements-registration-pharmaceuticals-human-use-considerations-general-principles-address-virus-and-vector-shedding_en.pdfEuropean Medicines Agency. Assessment Report: Zolgensma. International Non-Proprietary Name: Onasemnogene abeparvovec. Procedure No. EMEA/H/C/004750/0000. European Medicines Agency; 2020. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/zolgensma-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Zolgensma. International Non-Proprietary Name: Onasemnogene abeparvovec. Procedure No. EMEA/H/C/004750/0000. European Medicines Agency; 2020. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/zolgensma-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Luxturna. International Non-Proprietary Name: Voretigene neparvovec. Procedure No. EMEA/H/C/004451/0000. European Medicines Agency; 2019. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/luxturna-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Luxturna. International Non-Proprietary Name: Voretigene neparvovec. Procedure No. EMEA/H/C/004451/0000. European Medicines Agency; 2019. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/luxturna-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Upstaza. International Non-Proprietary Name: Eladocagene exuparvovec. Procedure No. EMEA/H/C/005352/0000. European Medicines Agency; 2022. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/upstaza-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Upstaza. International Non-Proprietary Name: Eladocagene exuparvovec. Procedure No. EMEA/H/C/005352/0000. European Medicines Agency; 2022. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/upstaza-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Roctavian. International Non-Proprietary Name: Valoctocogene roxaparvovec. Procedure No. EMEA/H/C/005830/0000. European Medicines Agency; 2022. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/roctavian-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Roctavian. International Non-Proprietary Name: Valoctocogene roxaparvovec. Procedure No. EMEA/H/C/005830/0000. European Medicines Agency; 2022. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/roctavian-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Glybera. International Nonproprietary Name: Alipogene tiparvovec. Procedure No. EMEA/H/C/002145. European Medicines Agency; 2012. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/glybera-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Glybera. International Nonproprietary Name: Alipogene tiparvovec. Procedure No. EMEA/H/C/002145. European Medicines Agency; 2012. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/glybera-epar-public-assessment-report_en.pdfSummary Basis for Regulatory Action - HEMGENIX. FDA; 2022. Accessed April 13, 2025. https://www.fda.gov/media/164094/download?attachmentSummary Basis for Regulatory Action - HEMGENIX. FDA; 2022. Accessed April 13, 2025. https://www.fda.gov/media/164094/download?attachmentSummary Basis for Regulatory Action – ELEVIDYS. FDA; 2023. Accessed April 13, 2025. https://www.fda.gov/media/169746/download?attachmentSummary Basis for Regulatory Action – ELEVIDYS. FDA; 2023. Accessed April 13, 2025. https://www.fda.gov/media/169746/download?attachmentSummary Basis for Regulatory Action. FDA; 2015. Accessed April 13, 2025. https://wayback.archive-it.org/7993/20190425013447/ https://www.fda.gov/downloads/BiologicsBloodVaccines/CellularGeneTherapyProducts/ApprovedProducts/UCM473103.pdfSummary Basis for Regulatory Action. FDA; 2015. Accessed April 13, 2025. https://wayback.archive-it.org/7993/20190425013447/ https://www.fda.gov/downloads/BiologicsBloodVaccines/CellularGeneTherapyProducts/ApprovedProducts/UCM473103.pdfМельникова Е.В., Рачинская О.А., Меркулов В.А. Высокотехнологические лекарственные препараты на основе онколитических вирусов (часть 1: разработка и регистрация в КНР). Ведомости Научного центра экспертизы средств медицинского применения. Регуляторные исследования и экспертиза лекарственных средств. 2021;11(3):148–159. https://doi.org/10.30895/1991-2919-2021-11-148-159Melnikova EV, Rachinskaya OA, Merkulov VA. Advanced therapy medicines based on oncolytic viruses (part I: development and authorisation of products in China). The Bulletin of the Scientific Centre for Expert Evaluation of Medicinal Products. 2021;11(3):148-159. (In Russ.). https://doi.org/10.30895/1991-2919-2021-11-148-159Коллегия Евразийской экономической комиссии. Решение от 26 ноября 2019 г. № 202 «Об утверждении Руководства по доклиническим исследованиям безопасности в целях проведения клинических исследований и регистрации лекарственных препаратов». Коллегия Евразийской экономической комиссии; 2019. Дата обращения: 13.04.2025. https://docs.eaeunion.org/upload/iblock/072/nzmd6i2fwbj90ndkm4riswwcz9ecgsa2/clcd_29112019_202_doc.pdfКоллегия Евразийской экономической комиссии. Решение от 26 ноября 2019 г. № 202 «Об утверждении Руководства по доклиническим исследованиям безопасности в целях проведения клинических исследований и регистрации лекарственных препаратов». Коллегия Евразийской экономической комиссии; 2019. Дата обращения: 13.04.2025. https://docs.eaeunion.org/upload/iblock/072/nzmd6i2fwbj90ndkm4riswwcz9ecgsa2/clcd_29112019_202_doc.pdfEuropean Medicines Agency. Assessment Report: Durveqtix. International non-proprietary name: Fidanacogene elaparvovec. Procedure No. EMEA/H/C/004774/0000. European Medicines Agency; 2024. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/durveqtix-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Durveqtix. International non-proprietary name: Fidanacogene elaparvovec. Procedure No. EMEA/H/C/004774/0000. European Medicines Agency; 2024. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/durveqtix-epar-public-assessment-report_en.pdfSummary Basis for Regulatory Action – ADSTILADRIN. FDA; 2022. Accessed April 13, 2025. https://www.fda.gov/media/164532/download?attachmentSummary Basis for Regulatory Action – ADSTILADRIN. FDA; 2022. Accessed April 13, 2025. https://www.fda.gov/media/164532/download?attachmentTsai HY, Hamilton A, Tinch E, et al. Genome wide association and genomic prediction for growth traits in juvenile farmed Atlantic salmon using a high density SNP array. BMC Genomics. 2015;16:969. PMID: 26582102. PMCID: PMC4652364. https://doi.org/10.1186/s12864-015-2117-9Tsai HY, Hamilton A, Tinch E, et al. Genome wide association and genomic prediction for growth traits in juvenile farmed Atlantic salmon using a high density SNP array. BMC Genomics. 2015;16:969. PMID: 26582102. PMCID: PMC4652364. https://doi.org/10.1186/s12864-015-2117-9Астапова О.В., Берчатова А.А. Генотерапевтические препараты: аспекты доклинического изучения безопасности. Безопасность и риск фармакотерапии. 2023;11(1):73–96. https://doi.org/10.30895/2312-7821-2023-11-1-329Astapova OV, Berchatova AA. Gene therapy medicinal products: non-clinical safety studies. Safety and Risk of Pharmacotherapy. 2023;11(1):73–96. (In Russ.). https://doi.org/10.30895/2312-7821-2023-11-1-329Donsante A, Miller DG, Li Y, et al. AAV vector integration sites in mouse hepatocellular carcinoma. Science. 2007;317(5837):477. PMID: 17656716. https://doi.org/10.1126/science.1142658Donsante A, Miller DG, Li Y, et al. AAV vector integration sites in mouse hepatocellular carcinoma. Science. 2007;317(5837):477. PMID: 17656716. https://doi.org/10.1126/science.1142658Dalwadi DA, Torrens L, Abril-Fornaguera J, et al. Liver injury increases the incidence of HCC following AAV gene therapy in mice. Mol Ther. 2021;29(2):680–690. PMID: 33554867. PMCID: PMC7854305. https://doi.org/10.1016/j.ymthe.2020.10.018Dalwadi DA, Torrens L, Abril-Fornaguera J, et al. Liver injury increases the incidence of HCC following AAV gene therapy in mice. Mol Ther. 2021;29(2):680–690. PMID: 33554867. PMCID: PMC7854305. https://doi.org/10.1016/j.ymthe.2020.10.018Chandler RJ, LaFave MC, Varshney GK, et al. Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy. J Clin Invest. 2015;125(2):870–880. PMID: 25607839. PMCID: PMC4319425. https://doi.org/10.1172/JCI79213Chandler RJ, LaFave MC, Varshney GK, et al. Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy. J Clin Invest. 2015;125(2):870–880. PMID: 25607839. PMCID: PMC4319425. https://doi.org/10.1172/JCI79213Nguyen GN, Everett JK, Kafle S, et al. A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells. Nat Biotechnol. 2021;39(1):47–55. PMID: 33199875. PMCID: PMC7855056. https://doi.org/10.1038/s41587-020-0741-7Nguyen GN, Everett JK, Kafle S, et al. A long-term study of AAV gene therapy in dogs with hemophilia A identifies clonal expansions of transduced liver cells. Nat Biotechnol. 2021;39(1):47–55. PMID: 33199875. PMCID: PMC7855056. https://doi.org/10.1038/s41587-020-0741-7Delytact Injection_Daiichi Sankyo Company, Limited_Review Report. PMDA; 2021. Accessed April 13, 2025. https://www.pmda.go.jp/files/000242808.pdfDelytact Injection_Daiichi Sankyo Company, Limited_Review Report. PMDA; 2021. Accessed April 13, 2025. https://www.pmda.go.jp/files/000242808.pdfCouto L, Parker A, Gordon JW. Direct exposure of mouse spermatozoa to very high concentrations of a serotype-2 adeno-associated virus gene therapy vector fails to lead to germ cell transduction. Hum Gene Ther. 2004;15(3):287–291. PMID: 15018737. https://doi.org/10.1089/104303404322886138Couto L, Parker A, Gordon JW. Direct exposure of mouse spermatozoa to very high concentrations of a serotype-2 adeno-associated virus gene therapy vector fails to lead to germ cell transduction. Hum Gene Ther. 2004;15(3):287–291. PMID: 15018737. https://doi.org/10.1089/104303404322886138Arruda VR, Fields PA, Milner R, et al. Lack of germline transmission of vector sequences following systemic administration of recombinant AAV-2 vector in males. Mol Ther. 2001;4(6):586–592. PMID: 11735343. https://doi.org/10.1006/mthe.2001.0491Arruda VR, Fields PA, Milner R, et al. Lack of germline transmission of vector sequences following systemic administration of recombinant AAV-2 vector in males. Mol Ther. 2001;4(6):586–592. PMID: 11735343. https://doi.org/10.1006/mthe.2001.0491Roehl HH, Leibbrandt ME, Greengard JS, et al. Analysis of testes and semen from rabbits treated by intravenous injection with a retroviral vector encoding the human factor VIII gene: no evidence of germ line transduction. Hum Gene Ther. 2000;11(18):2529–2540. PMID: 11119423. https://doi.org/10.1089/10430340050208000Roehl HH, Leibbrandt ME, Greengard JS, et al. Analysis of testes and semen from rabbits treated by intravenous injection with a retroviral vector encoding the human factor VIII gene: no evidence of germ line transduction. Hum Gene Ther. 2000;11(18):2529–2540. PMID: 11119423. https://doi.org/10.1089/10430340050208000European Medicines Agency. Assessment Report: Imlygic. International Non-Proprietary Name: Talimogene laherparepvec. Procedure No. EMEA/H/C/002771/0000European Medicines Agency; 2019. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/imlygic-epar-public-assessment-report_en.pdfEuropean Medicines Agency. Assessment Report: Imlygic. International Non-Proprietary Name: Talimogene laherparepvec. Procedure No. EMEA/H/C/002771/0000European Medicines Agency; 2019. Accessed April 13, 2025. https://www.ema.europa.eu/en/documents/assessment-report/imlygic-epar-public-assessment-report_en.pdf

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