Core C: PREVENT Program Pharmacokinetics and Pharmacodynamics Services Core

药代动力学和药效学（PK/PD）服务核心的目标是提供 集中支持项目2中临床前猕猴研究常见的PK和关键PD分析 项目3中的人体临床研究，从而确保项目研究的完整性和可比性 临床前和临床研究之间的差异。为此，我们将提供以下功能。 (1)我们将验证格里菲斯辛（GRFT）参比标准品的质量。一系列的生化反应， 生物物理学和病毒学分析将用于验证GRFT的质量。(2)我们将开发一个 通过免疫测定来验证GLP/GMP生产的GRFT的效力。在验证试验后， OCRP，其标准操作程序将转移到项目1和核心B。(3)我们会发现 并定量生物样品中的GRFT活性药物成分（API）和GRFT结合抗体， 用于支持项目2和3中PK/PD分析的液体。除了传统的基于 酶联免疫吸附测定，我们将开发基于表面等离子体共振的分析 检测低亲和力、快速解离抗体的方法。作为PK评估的一部分，我们将使用 HIV-1中和和细胞-细胞融合试验测定直肠液抗HIV-1能力 样品(4)我们将分析GRFT凝胶治疗对粘膜环境的影响，以支持 体内安全性评估。为此，我们将采用系统生物学方法， 揭示了直肠施用GRFT后可能发生的粘膜环境的变化， GRFT组合凝胶。在粘膜和淋巴结中诱导先天性和适应性免疫应答， 将使用蛋白质组学技术评估全身免疫区室。我们还将高度利用 敏感的免疫组织化学技术，以比较HIV靶细胞的数量和位置， 直肠组织中的上皮连接蛋白。此外，整个直肠的微生物群结构 GRFT和组合凝胶处理的过程将通过条形码16 S rRNA基因进行监测 测序以揭示杀微生物剂制剂对直肠的分类结构的影响 微生物群

The objective of the Pharmacokinetics and Pharmacodynamics (PK/PD) Services Core will be to provide centralized support for the PK and key PD analyses common to preclinical macaque studies in Project 2 and human clinical studies in Project 3, thereby ensuring Program's research integrity and comparability between the preclinical and clinical studies proposed. To this end, we will provide the following functions. (1) We will validate the quality of a griffithsin (GRFT) reference standard. A series of biochemical, biophysical and virological assays will be used to validate the quality of GRFT. (2) We will develop an immunoassay to validate the potency of GLP/GMP-manufactured GRFT. Upon validation of the assay at OCRP, its standard operating procedure will be transferred to Project 1 and Core B. (3) We will detect and quantify GRFT active pharmaceutical ingredient (API) and GRFT-binding antibodies in biological fluids to support PK/PD analysis in Projects 2 and 3. In addition to conventional methods based on enzyme-linked immunosorbent assays, we will develop surface plasmon resonance-based analytical methods to detect low-affinity, rapidly dissociating antibodies. As part of PK assessment, we will use HIV-1 neutralization and cell-cell fusion assays to measure the anti-HIV-1 capacity of rectal fluid samples. (4) We will analyze the impact of GRFT gel treatment on the mucosal environment to support in vivo safety assessment. To this end, we will employ a systems biology approach to comprehensively reveal the changes in the mucosal environment that may occur upon rectal administration of GRFT and GRFT combination gels. The induction of innate and adaptive immune responses in the mucosal and systemic immune compartments will be assessed using proteomic techniques. We will also use highly sensitive immunohistochemistry techniques to compare the number and location of HIV target cells, and epithelial junction proteins in rectal tissue. Additionally, the structure of rectal microbiota throughout the course of GRFT and combination gel treatment will be monitored by barcoded 16S rRNA genes sequencing to reveal the effect of microbicides formulation on the taxonomic structure of the rectal microbiota.