Institute of Dentistry, School of Medicine Medical Sciences & Nutrition, University of Aberdeen, Aberdeen AB25 2ZR, U.K.
Laboratory of Molecular Microbiology and Biotechnology, Department of Medical Biotechnologies, University of Siena, Siena 53100, Italy.
Mol Pharm. 2020 Mar 2;17(3):852-864. doi: 10.1021/acs.molpharmaceut.9b01121. Epub 2020 Feb 20.
Clinical trials have demonstrated partial protection against HIV-1 infection by vaginal microbicide formulations based on antiretroviral (ARV) drugs. Improved formulations that will maintain sustained drug concentrations at viral target sites in the cervicovaginal mucosa are needed. We have previously demonstrated that treatment of cervicovaginal cell lines with ARV drugs can alter gene expression of drug transporters, suggesting that the mucosal disposition of ARV drugs delivered vaginally can be modulated by drug transporters. This study aimed to investigate modulation of drug transporter expression in a nonhuman primate model by tenofovir and darunavir released from film formulations. Cervicovaginal tissues were collected from drug-naïve macaques and from macaques vaginally treated with film formulations of tenofovir or darunavir. Drug release in vaginal fluid as well as drug absorption in cervicovaginal tissues and lymph nodes were verified by mass spectrometry. The effects of exposure to drugs on the expression of transporters relevant to ARV drugs were evaluated by quantitative PCR. We showed expression in cervicovaginal tissue of drug-naïve macaques of transporters important for distribution of ARV drugs, albeit at lower levels compared to human tissue for key transporters including P-glycoprotein. Concentrations of tenofovir and darunavir well above the EC values determined were detected in vaginal fluid and vaginal tissues of macaques treated with drug-dissolving films over 24 h and were also comparable to those shown previously to modulate drug transporter expression. Accordingly, Multidrug Resistance associated Protein 2 (MRP2) in cervicovaginal tissue was upregulated by both tenofovir and darunavir. The two drugs also differentially induced and/or inhibited expression of key uptake transporters for reverse transcriptase inhibitors and protease inhibitors. The lower expression of key transporters in macaques may result in increased retention of ARV drugs at the simian cervicovaginal mucosa compared to the human mucosa and has implications for translation of preclinical data. Modulation of drug transporter expression by tenofovir and darunavir points to the potential benefit of MRP2 inhibition to increase ARV drug penetration through the cervicovaginal epithelium.
临床试验已经证明，基于抗逆转录病毒（ARV）药物的阴道杀微生物剂配方可以提供针对 HIV-1 感染的部分保护。需要开发能够在宫颈阴道黏膜中的病毒靶位维持持续药物浓度的改良配方。我们之前已经证明，用 ARV 药物处理宫颈阴道细胞系可以改变药物转运体的基因表达，表明阴道给予的 ARV 药物的黏膜处置可以通过药物转运体来调节。本研究旨在通过从薄膜制剂中释放的替诺福韦和达芦那韦来研究非人类灵长类动物模型中药物转运体表达的调节。从未经药物处理的猕猴和阴道用薄膜制剂处理的猕猴中收集宫颈阴道组织。通过质谱法验证了阴道液中的药物释放以及宫颈阴道组织和淋巴结中的药物吸收。通过定量 PCR 评估了暴露于药物对与 ARV 药物相关的转运体表达的影响。我们在未经药物处理的猕猴的宫颈阴道组织中显示了与 ARV 药物分布相关的重要转运体的表达，尽管与关键转运体（包括 P-糖蛋白）的人类组织相比，其表达水平较低。在 24 小时内用药物溶解膜处理的猕猴的阴道液和阴道组织中检测到远高于 EC 值的替诺福韦和达芦那韦的浓度，并且与先前显示可调节药物转运体表达的浓度相当。因此，多药耐药相关蛋白 2（MRP2）在宫颈阴道组织中被替诺福韦和达芦那韦上调。这两种药物还分别诱导和/或抑制了逆转录酶抑制剂和蛋白酶抑制剂的关键摄取转运体的表达。与人类黏膜相比，猕猴中关键转运体的较低表达可能导致 ARV 药物在猴宫颈阴道黏膜中的保留增加，这对转化临床前数据具有重要意义。替诺福韦和达芦那韦对药物转运体表达的调节表明，MRP2 抑制可能有助于增加 ARV 药物穿透宫颈阴道上皮的渗透。
