Core C: PREVENT Program Pharmacokinetics and Pharmacodynamics Services Core

核心 C：预防计划药代动力学和药效学服务核心

• 批准号: 8769376
• 负责人: Nobuyuki Matoba
• 金额: $ 55.17万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8769376
• 关键词: Adverse effects; Affinity; Anti-HIV Agents; Antibodies; Binding; Biochemical; Biological; Biological Assay; Biological Markers; Blood specimen; Body Fluids; Cell Line; Cell fusion; Cells; Clinical Research; Complement; Complex; Cyclic GMP; Detection; Development; Drug Formulations; Drug Kinetics; Ensure; Environment; Enzyme-Linked Immunosorbent Assay; Epithelial Cell Junction; Evaluation; Fluorescence; Gel; Gene Expression Profiling; Giant Cells; Glycoproteins; HIV; HIV Envelope Protein gp120; HIV-1; Hela Cells; High Pressure Liquid Chromatography; Histopathology; Human; Human Herpesvirus 2; IgE; Immune; Immune response; Immunoassay; Immunoblotting; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Immunohistochemistry; Lead; Liquid substance; Location; Macaca; Macaca mulatta; Measures; Methods; Molecular Sieve Chromatography; Monitor; Mucous Membrane; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Prevention; Procedures; Proteins; Proteomics; Quality Control; Reagent; Rectal Administration; Reference Standards; Reporter Genes; Research; Safety; Sampling; Series; Services; Structure; Surface Plasmon Resonance; Systems Biology; Techniques; Technology Transfer; Testing; Tissues; Toxicology; Validation; Viral; analytical method; base; immunogenicity; in vivo; microbicide; pre-clinical; pre-clinical research; preclinical study; programs; rRNA Genes; rectal; rectal microbicide

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.

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 所提出的临床前研究和临床研究之间的关系。为此，我们将提供以下功能。 (1) 我们将验证格里菲辛 (GRFT) 参考标准品的质量。一系列生化、 生物物理和病毒学检测将用于验证 GRFT 的质量。 (2) 我们将开发一个 免疫测定法验证 GLP/GMP 制造的 GRFT 的效力。测定验证后 OCRP，其标准操作流程将转移到项目1和核心B。 (3)我们将检测 并量化生物中的 GRFT 活性药物成分 (API) 和 GRFT 结合抗体 流体支持项目 2 和 3 中的 PK/PD 分析。除了基于 酶联免疫吸附测定，我们将开发基于表面等离子共振的分析 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 中和和细胞融合试验可测量直肠液的抗 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来全面 揭示直肠给予 GRFT 后可能发生的粘膜环境变化 GRFT 组合凝胶。在粘膜和粘膜中诱导先天性和适应性免疫反应 将使用蛋白质组学技术评估全身免疫区室。我们也将高度使用 敏感的免疫组织化学技术来比较 HIV 靶细胞的数量和位置，以及 直肠组织中的上皮连接蛋白。此外，整个直肠微生物群的结构 GRFT 和组合凝胶治疗的过程将通过带条形码的 16S rRNA 基因进行监测 测序揭示杀菌剂配方对直肠分类结构的影响 微生物群。