Drug at the Right Place & Concentration: Optimizing Combination Vaginal Ring

项目总结(见说明)：~ 生物分析核心的目标是为项目1和2中建议的临床前研究以及项目3中建议的第一阶段临床研究开发和应用有效的分析方法来确定药物水平。核心还将根据良好的制造规范(GMP)制备阴道内环(IVR)，以提供富马酸替诺福韦(TDF)，这是替诺福韦(TFV)的一种更有效的前体药物，具有极好的临床前安全性，用于第一阶段药代动力学(PK)研究。该中心已经开发和优化了用于体外研究的生物分析方法，以检测TDF和相关代谢物，并将扩大这项工作，并根据ICH指南验证Tenofovir(TFV)的替代有效前体药物GS7340以及具有逆转录酶和进入抑制活性的嘧啶二酮和马拉韦罗(MVC)的定量方法。CORE还将验证检测细胞内2‘脱氧三磷酸脱氧腺苷(DATP)浓度的方法，该脱氧腺苷是替诺福韦二磷酸(TFV-DP)的天然底物和竞争类似物。共同研究人员Craig Hendrix博士是临床试验中血液、外周血单个核细胞(PBMC)、宫颈阴道灌洗液(CVL)和宫颈阴道(CV)抽吸物、直肠海绵以及阴道和宫颈活检组织中TFV和TFV-二磷酸(活性代谢物)检测方法的领导者，他将扩展其实验室建立的经临床验证的UPLC/MS/MS方法，并建立量化临床样本中TDF的检测方法。 一旦符合ICH和FDA的标准，这些检测方法(连同已建立的TFV和TFV-DP检测方法)将被应用于项目3中第4年和第5年提出的第一阶段前期PK研究中获得的样品的分析。这将包括测量CV抽吸物、宫颈和阴道活检、直肠海绵和血液中的TDF和适当的代谢物。CORE将利用粒子科学公司正在进行的工作，开发TDF聚氨酯阴道内环，进行监管测试，并提交IND，并将支持产品的开发，并为第一阶段前阴道环研究提供临床用品。

PROJECT SUMMARY (See instructions): ~~~~ The objectives of the Bioanalytical Core are to develop and apply validated analytical methods for determination of drug levels for preclinical studies proposed in Projects 1 and 2 and for a pre-Phase I clinical study proposed in Project 3. The Core will also prepare intravaginal rings (IVRs) under Good Manufacturing Practice (GMP) to deliver tenofovir disoproxil fumarate (TDF), a more potent prodrug of tenofovir (TFV) with an excellent preclinical safety profile for a pre-Phase I pharmacokinetic (PK) study. The Core has developed and optimized bioanalytical methods to detect TDF and related metabolites for in vitro studies and will expand this work and validate as per ICH guidelines methods for quantification of GS7340, an alternative potent prodrug of tenofovir (TFV), as well as for IQP-0528, a pyrimidinedione with reverse transcriptase and entry inhibitory activity and maraviroc (MVC). The Core will also validate methods to detect intracellular concentrations of 2'deoxyadenosine triphosphate (dATP), the natural substrate and competitive analogue of tenofovir diphosphate (TFV-DP). Dr. Craig Hendrix, co-investigator, who is a leader in the development and validation of assays to measure TFV and TFV-diphosphate (the active metabolite) in blood, peripheral blood mononuclear cells (PBMCs), cervicovaginal lavages (CVL) and cervicovaginal (CV) aspirates, rectal sponges, and vaginal and cervical biopsy tissue for clinical trials, will expand the clinically validated UPLC/MS/MS methods established in his laboratory and develop assays to quantify TDF in clinical samples. Once validated to ICH and FDA standards, these assays (along with established TFV and TFV-DP assays) will be applied to the analysis of samples obtained in a pre-Phase I PK study proposed in Years 4 and 5 in Project 3. This will include measurements of TDF and appropriate metabolites in CV aspirates, cervical and vaginal biopsies, rectal sponges, and blood. The Core will leverage the ongoing work at Particle Sciences in the development of the TDF polyurethane intravaginal ring, regulatory testing, and filing of an IND and will support the development of the product and provide clinical supplies for the pre-Phase I vaginal ring study.