Project 2: Safety Evaluation of Combination Antiretroviral Intravaginal Rings

项目概要：项目2 安全性证明是局部非疫苗生物医学制剂开发的核心组成部分， 预防（nBP）候选人预防性传播艾滋病毒。当口服FDA- 批准的药物被重新配制用于阴道局部给药，安全性研究涉及一般的 一种或多种动物种属的毒理学和刺激/炎症，沿着 对阴道菌群的影响，通常在猕猴中。这些既定的研究并没有充分解决所有 阴道给药抗逆转录病毒（ARV）组合的重要安全性方面，特别是那些 与通过使用产品增加艾滋病毒感染风险有关，这可能会抵消药物的保护作用， 方面的影响.项目2的目的是评估阴道环（IVRs）给药的安全性/局部毒性 核心B中配制的抗逆转录病毒药物复方制剂。在目标1中，将在体外筛选9种初始ARV IVR制剂， 高通量阴道上皮细胞（VEC）培养系统，以测量药物组合的影响， 给药：生理和健康;先天免疫刺激;细胞凋亡和炎症;和共生 使用定植有复杂阴道微生物群的VEC来评估微生物活力和健康。初步安全性 也将在第一轮小鼠和绵羊研究中建立。这些安全性和有效性的结果 （项目3）的结果将用于选择三名抗逆转录病毒IVR候选人进行更深入的第二轮评估。 根据Aim 2，在绵羊模型中使用先进的体内高分辨率成像以及 传统的组织学和阴道镜检查这些IVR的局部毒性将在目标3中进一步评估，使用 通过测量阴道上皮细胞中炎性细胞因子的上调， IVR对肠道菌群的影响。后者将通过以下创新组合来实现： 分子技术，以获得不依赖于培养的微生物群落组成和微观 技术来表征阴道和IVRs表面的群落结构， 特别强调微生物生物膜。局部毒性的类似评估将应用于（预- I期）使用我们的主要候选药物IVR三联抗逆转录病毒药物联合治疗的女性探索性临床试验（目的3）。 成功完成这些安全性研究将为选择候选药物提供必要的数据 并将成为在该制剂向IPCP后过渡过程中的IND赋能活动的基石 临床研究。

PROJECT SUMMARY: PROJECT 2 The demonstration of safety is a central component in the development of topical non-vaccine biomedical prevention (nBP) candidates for the prevention of sexual HIV transmission. When orally administered FDA- approved drugs are reformulated for topical delivery to the vagina, safety studies have involved general toxicology and irritation/inflammation in one or more animal species, along with a rudimentary assessment of the effect on vaginal microflora, usually in macaques. These established studies do not adequately address all important safety aspects of antiretroviral (ARV) combinations administered to the vagina, particularly those relating to increased risk of HIV acquisition through product use that could counteract the drugs' protective effects. The objective of Project 2 is to assess the safety/local toxicity of intravaginal rings (IVRs) delivering ARV combinations formulated in Core B. In Aim 1, nine initial ARV IVR formulations will be screened ex vivo in a high-throughput vaginal epithelial cell (VEC) culture system to measure the impact of drug combinations and dosing on: physiology and health; innate immune stimulation; apoptosis and inflammation; and commensal microorganism viability and health using VECs colonized with complex vaginal microbiota. Preliminary safety also will be established in a first round of mouse and sheep studies. Results from these safety and efficacy (Project 3) outcomes will be used to select three ARV IVR candidates for more in-depth, second round of safety evaluation under Aim 2 in a sheep model using advanced, in vivo, high-resolution imaging together with traditional histology and colposcopy. The local toxicity of these IVRs will be assessed further in Aim 3 using pig-tailed macaques by measuring the upregulation of inflammatory cytokines in the vaginal epithelium and the impact of the IVRs on commensal microflora. The latter will be achieved using an innovative combination of molecular techniques to obtain culture-independent microbial community composition and microscopic techniques to characterize the community structure in the vaginal tract and on the surface of the IVRs with a special emphasis on microbial biofilms. An analogous assessment of local toxicity will be applied in a (pre- Phase I) Exploratory Clinical Trial in women using our lead candidate IVR triple ARV combination (Aim 3). Successful completion of these safety studies will provide the necessary data for selection of a lead candidate and will form a cornerstone of the IND-enabling activities in the transition of this formulation to post-IPCP clinical studies.