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 in vivo 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 EC50 values determined in vitro 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.

中文翻译：

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膜释放替诺福韦和达那那韦用于局部预防HIV-1的宫颈阴道药物转运蛋白的体内调节和组织分布。

临床试验表明，基于抗逆转录病毒（ARV）药物的阴道杀菌剂配方可部分保护HIV-1免受感染。需要改进的制剂以在宫颈阴道粘膜的病毒靶位点上维持持续的药物浓度。先前我们已经证明用ARV药物治疗宫颈阴道细胞系可以改变药物转运蛋白的基因表达，这表明药物转运蛋白可以调节阴道输送的ARV药物的粘膜处置。这项研究旨在研究通过薄膜制剂释放的替诺福韦和达那那韦在非人类灵长类动物模型中体内对药物转运蛋白表达的调节。从无毒品的猕猴和经替诺福韦或达鲁纳韦的薄膜制剂经阴道处理的猕猴中收集宫颈阴道组织。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评估暴露于药物对与抗逆转录病毒药物有关的转运蛋白表达的影响。我们显示了在初次免疫的猕猴的子宫颈阴道组织中的表达，这些猕猴对ARV药物的分配很重要，尽管与包括P-糖蛋白在内的关键转运蛋白的人体组织相比，其水平较低。在经过药物溶解膜处理超过24小时的猕猴的阴道液和阴道组织中，检测到的tenofovir和darunavir的浓度远高于体外测定的EC50值，并且与以前显示的调节药物转运蛋白表达的浓度相当。因此，替诺福韦和达那那韦均上调了宫颈阴道组织中的多药耐药相关蛋白2（MRP2）。两种药物还差异诱导和/或抑制逆转录酶抑制剂和蛋白酶抑制剂的关键摄取转运蛋白的表达。与人粘膜相比，猕猴中关键转运蛋白的较低表达可能导致ARV药物在猿猴宫颈阴道粘膜中的保留增加，并且对临床前数据的翻译具有影响。替诺福韦和达那那韦对药物转运蛋白表达的调节表明，抑制MRP2可能增加ARV药物通过子宫颈上皮的渗透。两种药物还差异诱导和/或抑制逆转录酶抑制剂和蛋白酶抑制剂的关键摄取转运蛋白的表达。与人粘膜相比，猕猴中关键转运蛋白的较低表达可能导致ARV药物在猿猴宫颈阴道粘膜中的保留增加，并且对临床前数据的翻译具有影响。替诺福韦和达那那韦对药物转运蛋白表达的调节表明，抑制MRP2可能增加ARV药物通过子宫颈上皮的渗透。两种药物还差异诱导和/或抑制逆转录酶抑制剂和蛋白酶抑制剂的关键摄取转运蛋白的表达。与人粘膜相比，猕猴中关键转运蛋白的较低表达可能导致ARV药物在猿猴宫颈阴道粘膜中的保留增加，并且对临床前数据的翻译具有影响。替诺福韦和达那那韦对药物转运蛋白表达的调节表明，抑制MRP2可能增加ARV药物通过子宫颈上皮的渗透。