Определение паклитаксела и ритонавира в плазме крови и органах крыс методом высокоэффективной жидкостной хроматографии

Паклитаксел (PTX) — цитотоксический противоопухолевый агент из группы таксанов, низкая биодоступность и тканевое биораспределение которого могут быть изменены в присутствии ингибиторов ферментов печени и транспортных белков, например, ритонавира (RTV) — антиретровирусного агента, применяемого в терапии ВИЧ-инфекции. Разработана и валидирована методика одновременного определения PTX и RTV в плазме крови, гомогенатах печени, мозга и тестикул крыс методом обращенно-фазовой высокоэффективной жидкостной хроматографии (ВЭЖХ) с диодно-матричным детектированием. Пробоподготовка образцов включала жидкость-жидкостную экстракцию с последующим упариванием экстракта и повторным диспергированием в подвижной фазе с дополнительной очисткой. Диапазон определяемых содержаний PTX составляет 100 – 9090 нг/мл в плазме крови и 25 – 9090 нг/г в гомогенатах органов, RTV — 250 – 9090 нг/г во всех объектах анализа. Предел определения (ПО) PTX в плазме — 100 нг/мл, RTV — 250 нг/мл, в органах — 25 и 250 нг/г соответственно. Разработанная методика показала высокую точность (относительная ошибка Δ < 12 %) и прецизионность (относительное стандартное отклонение RSD < 14 %) во всем диапазоне концентраций; степень извлечения PTX и RTV составила >80 % для всех биологических объектов. Предложенная методика может быть использована при изучении фармакокинетики данной комбинации лекарственных веществ в биологических матрицах.

1. Škubník J., Pavlíčková V., Ruml T., Rimpelová S. Current perspectives on taxanes: focus on their bioactivity, delivery and combination therapy / Plants. 2021. Vol. 10. No. 3. P. 1 – 35. DOI: 10.3390/plants10030569

2. Jibodh R. A., Lagas J. S., Nuijen B., et al. Taxanes: old drugs, new oral formulations / Eur. J. Pharmacol. 2013. Vol. 717. Nos. 1 – 3. P. 40 – 46. DOI: 10.1016/j.ejphar.2013.02.058

3. Hendrikx J. J. M. A., Lagas J. S., Rosing H., et al. P-glycoprotein and cytochrome P450 3A act together in restricting the oral bioavailability of paclitaxel / Int. J. Cancer. 2013. Vol. 132. No. 10. P. 2439 – 2447. DOI: 10.1002/ijc.27912

4. Lee C. K., Choi J. S. Effects of silibinin, inhibitor of CYP3A4 and P-glycoprotein in vitro, on the pharmacokinetics of paclitaxel after oral and intravenous administration in rats / Pharmacology. 2010. Vol. 85. No. 6. P. 350 – 356. DOI: 10.1159/000312690

5. Pereira M., Vale N. Ritonavir’s evolving role: a journey from antiretroviral therapy to broader medical applications / Curr. Oncol. 2024. Vol. 31. No. 10. P. 6032 – 6049. DOI: 10.3390/curroncol31100450

6. Loos N. H. C., Beijnen J. H., Schinkel A. H. The mechanism-based inactivation of CYP3A4 by ritonavir: what mechanism? / Int. J. Mol. Sci. 2022. Vol. 23. No. 17. DOI: 10.3390/ijms23179866

7. Hendrikx J. J. M. A., Lages J. S., Wagenaar E., et al. Oral co-administration of elacridar and ritonavir enhances plasma levels of oral paclitaxel and docetaxel without affecting relative brain accumulation / Br. J. Cancer. 2014. Vol. 110. No. 11. P. 2669 – 2676. DOI: 10.1038/bjc.2014.222

8. van Eijk M., Yu H., Sawicki E., et al. Development of a population pharmacokinetic/pharmacodynamic model for various oral paclitaxel formulations co-administered with ritonavir and thrombospondin-1 based on data from early phase clinical studies / Cancer Chemother. Pharmacol. 2022. Vol. 90. No. 1. P. 71 – 82. DOI: 10.1007/s00280-022-04445-z

9. de Weger V. A., Vermunt M. A. C., Stuurman F. E., et al. A phase 1 dose-escalation study of low-dose metronomic treatment with novel oral paclitaxel formulations in combination with ritonavir in patients with advanced solid tumors / Clin. Pharmacol. Drug Dev. 2021. Vol. 10. No. 6. P. 607 – 621. DOI: 10.1002/cpdd.880

10. Murdoch I. G., Jacobs T. G., Nieuwenhuize R. S., et al. Ritonavir-boosted antiretroviral therapy with paclitaxel: will it lead to boosted toxicity? / AIDS. 2022. Vol. 36. No. 2. P. 322 – 323. DOI: 10.1097/qad.0000000000003115

11. Rauschenbach L., Wieland A., Reinartz R., et al. Drug repositioning of antiretroviral ritonavir for combinatorial therapy in glioblastoma / Eur. J. Cancer. 2020. Vol. 140. P. 130 – 139. DOI: 10.1016/j.ejca.2020.09.017

12. van der Putten E., Wosikowski K., Beijnen J. H., et al. Ritonavir reverses resistance to docetaxel and cabazitaxel in prostate cancer cells with acquired resistance to docetaxel / Cancer Drug Resist. 2024. Vol. 7. DOI: 10.20517/cdr.2023.136

13. Hendrikx J. J. M. A., Hillebrand M. J. X., Thijssen B., et al. A sensitive combined assay for the quantification of paclitaxel, docetaxel and ritonavir in human plasma using liquid chromatography coupled with tandem mass spectrometry / J. Chromatogr B. 2011. Vol. 879. No. 28. P. 2984 – 2990. DOI: 10.1016/j.jchromb.2011.08.034

14. Hendrikx J. J. M. A., Rosing H., Schinkel A. H., et al. Combined quantification of paclitaxel, docetaxel and ritonavir in human feces and urine using LC-MS/MS / Biomed. Chromatogr. 2014. Vol. 28. No. 2. P. 302 – 310. DOI: 10.1002/bmc.3021

15. Gote V., Deep Sharma A., Pal D. Hyaluronic acid-targeted stimuli-sensitive nanomicelles co-encapsulating paclitaxel and ritonavir to overcome multi-drug resistance in metastatic breast cancer and triple-negative breast cancer cells / Int. J. Mol. Sci. 2021. Vol. 22. No. 3. P. 1 – 35. DOI: 10.3390/ijms22031257

16. Khan I., Iqbal Z., Khan A., et al. A simple, rapid and sensitive RP-HPLC-UV method for the simultaneous determination of sorafenib & paclitaxel in plasma and pharmaceutical dosage forms: application to pharmacokinetic study / J. Chromatogr. B. 2016. Vol. 1033 – 1034. P. 261 – 270. DOI: 10.1016/j.jchromb.2016.08.029

17. Wang L. Z., Ho P. C., Lee H. S., et al. Quantitation of paclitaxel in micro-sample rat plasma by a sensitive reversed-phase HPLC assay / J. Pharm. Biomed. Anal. 2003. Vol. 31. No. 2. P. 283 – 289. DOI: 10.1016/s0731-7085(02)00611-8

18. Kim S. C., Yu J., Lee J. W., et al. Sensitive HPLC method for quantitation of paclitaxel (Genexol®) in biological samples with application to preclinical pharmacokinetics and biodistribution / J. Pharm. Biomed. Anal. 2005. Vol. 39. Nos. 1 – 2. P. 170 – 176. DOI: 10.1016/j.jpba.2005.02.023

19. Kumbhar P. S., Siwate S. K., Mali U. G., et al. Development and validation of RP-HPLC method for simultaneous estimation of docetaxel and ritonavir in PLGA nanoparticles / Ann. Pharm. Fr. 2020. Vol. 78. No. 5. P. 398 – 407. DOI: 10.1016/j.pharma.2020.07.004

20. Verbesselt R., Van Wijngaerden E., de Hoon J. Simultaneous determination of 8 HIV protease inhibitors in human plasma by isocratic high-performance liquid chromatography with combined use of UV and fluorescence detection: amprenavir, indinavir, atazanavir, ritonavir, lopinavir, saquinavir, nelfinavir and M8-nelfinavir metabolite / J. Chromatogr. B. 2007. Vol. 845. No. 1. P. 51 – 60. DOI: 10.1016/j.jchromb.2006.07.068

21. Soyinka J. O., Onyeji C. O., Omoruyi S. I. Simultaneous liquid chromatographic analysis of ritonavir, quinine and 3-hydroxyquinine in human plasma / J. Chromatogr. B. 2009. Vol. 877. No. 4. P. 441 – 445. DOI: 10.1016/j.jchromb.2008.12.045

22. G.Ph.A.1.1.0012. Validation of Analytical Procedures / State Pharmacopoeia of the Russian Federation (XV ed. 2023) [in Russian].

23. ICH guideline M10 on bioanalytical method validation and study sample analysis. 2022.

24. Lian H., Sun J., Zhang T. A rapid and sensitive determination of paclitaxel in rat plasma by UPLC-MS/MS method: application to a pharmacokinetic study / Asian. J. Pharm. Sci. 2013. Vol. 8. No. 3. P. 199 – 205. DOI: 10.1016/j.ajps.2013.07.026

25. Gaïes E., Jebabli N., Ben Said D., et al. Development and validation method for estimation paclitaxel in plasma / La Tunise medicale. 2020. Vol. 98. No. 6. P. 496 – 500.

26. Ph.A.2.1.0563. Ritonavir / State Pharmacopoeia of the Russian Federation (XV ed. 2023) [in Russian].

27. Ph.A.2.1.0544. Paclitaxel / State Pharmacopoeia of the Russian Federation (XV ed. 2023) [in Russian].

28. Mittal A., Chitkara D., Kumar N. HPLC method for the determination of carboplatin and paclitaxel with cremophor EL in an amphiphilic polymer matrix / J. Chromatogr. B. 2007. Vol. 855. No. 2. P. 211 – 219. DOI: 10.1016/j.jchromb.2007.05.005

29. Rezazadeh M., Emami J., Mostafavi A., et al. A rapid and sensitive HPLC method for quantitation of paclitaxel in biological samples using liquid-liquid extraction and UV detection: application to pharmacokinetics and tissues distribution study of paclitaxel loaded targeted polymeric micelles in tumor bearing mice / J. Pharm. Pharm. Sci. 2015. Vol. 18. No. 5. P. 647 – 660. DOI: 10.18433/j3rp6z

30. Guo W., Johnson J. L., Khan S., et al. Paclitaxel quantification in mouse plasma and tissues containing liposome-entrapped paclitaxel by liquid chromatography-tandem mass spectrometry: application to a pharmacokinetics study / Anal. Biochem. 2005. Vol. 336. No. 2. P. 213 – 220. DOI: 10.1016/j.ab.2004.09.046

31. Yonemoto H., Ogino S., Nakashima M. N., et al. Determination of paclitaxel in human and rat blood samples after administration of low dose paclitaxel by HPLC-UV detection / Biomed. Chromatogr. 2007. Vol. 21. No. 3. P. 310 – 317. DOI: 10.1002/bmc.759

32. Hoetelmans R. M. W., van Essenberg M., Profijt M., et al. High-performance liquid chromatographic determination of ritonavir in human plasma, cerebrospinal fluid and saliva / J. Chromatogr. B. 1998. Vol. 705. No. 1. P. 119 – 126. DOI: 10.1016/s0378-4347(97)00500-8

33. Keil K., Frerichs V. A., DiFrancesco R., Marse G., et al. Reverse phase high-performance liquid chromatography method for the analysis of amprenavir, efavirenz, indinavir, lopinavir, nelfinavir and its active metabolite (M8), ritonavir, and saquinavir in heparinized human plasma / Ther. Drug. Monit. 2003. Vol. 25. No. 3. P. 340 – 346. DOI: 10.1097/00007691-200306000-00015

34. Vaishampayan U. N., Kessen M., Dreicer R., et al. A global phase II randomized trial comparing oral taxane ModraDoc006/r to intravenous docetaxel in metastatic castration resistant prostate cancer / Eur. J. Cancer. 2024. Vol. 202. 114007. DOI: 10.1016/j.ejca.2024.114007