RT Inhibitor CSIC and Entry Inhibitor Retrocyclin RC101 as Microbicides

这份U19合作协议拨款申请旨在进行临床前研究，以优化由HIV-1进入抑制剂逆转录病毒RC101和非核苷逆转录酶抑制剂(NNRTI)5-氯-3-苯磺酰吲哚-2-甲酰胺(CSIC)组成的组合杀菌剂。我们的目标是开发一种杀微生物剂，在诱导最小的病毒抵抗力的同时，对无细胞和细胞相关病毒具有强大的抗HIV活性，对宫颈和阴道组织无毒和非炎性，并安全有效地对抗猴子的阴道SIV挑战。将根据四个项目和两个核心之间高度互动的方法，选择具有这些特性的RC-101和CSIC的最佳组合。我们将共同采用一系列体外和体外实验室的有效性和毒性测试，在猴子模型上的体内安全性和有效性测试，并研究优化我们化合物的硅橡胶环配方。这笔赠款申请的4个项目是：1)开发CSIC作为杀微生物剂(项目1，Parniak博士)；2)RC-101作为阴道内抗HIV-1局部杀菌剂(项目2，Cole博士)；3)在宫颈组织培养中测试候选杀菌剂的抗病毒活性和毒性(项目3，Gupta博士)；以及4)评价CSIC和RC-101在猴子身上的毒性和疗效(项目4，马克思博士)。这四个项目的科学研究将由行政核心(核心A，古普塔博士)和一个配方核心(核心B，Rohan博士和Smith)提供支持。汤姆·史密斯博士(Auritec PharmPharmticals)是这项U19计划的行业合作伙伴，他将带来控释微丸技术，以开发以RC-101和CSIC为基础的杀菌剂在猴子体内的阴道环状递送。在设计拟议的局部杀微生物剂计划的范围时，我们专注于利用体外研究、组织外植体和动物模型以及扩展和创新的器官培养模型来广泛评估候选杀菌剂的安全性和毒性，以研究在常见性传播感染存在的情况下杀菌剂的抗病毒活性。我们的杀微生物剂开发的多学科方法通过一种受控释放环配方产品针对HIV-1的两个不同复制位置(进入和反转录)，利用一种新型的体外器官培养来测试整个宫颈阴道粘膜的毒性、炎症和抗病毒活性，并应用猕猴模型来研究毒性和有效性。因此，这项拟议的研究将为开发用于预防艾滋病毒-1传播及其对妇女的后遗症的杀微生物剂提供新的战略。

This U19 Cooperative Agreement grant application is designed to perform preclinical studies to optimize a combination microbicide composed of the HIV-1 entry inhibitor retrocyclin RC101 and the nonnucleoside reverse transcriptase inhibitor (NNRTI) 5-chloro-3-phenylsulfonylindole-2-carboxamide (CSIC). Our goal is to develop a microbicide that will have potent anti-HIV activity against cell-free and cell-associated virus while inducing minimal viral resistance, be non-toxic and non-inflammatory to cervical and vaginal tissues and be safe and effective against vaginal SHIV challenge in monkeys. An optimal combination of RC-101 and CSIC with these properties will be chosen based on a highly interactive approach between four projects and two cores. Together, we will employ an array of in vitro and ex vivo laboratory tests of efficacy and toxicity, in vivo safety and efficacy tests in monkey models, and studies to optimize a silicone elastomer ring formulation of our compounds. The 4 projects of this grant application are: 1) Development of CSIC as a microbicide (Project 1, Dr. Parniak), 2) RC-101 as an intravaginal anti-HIV-1 topical microbicide (Project 2, Dr. Cole), 3) Testing candidate microbicides for antiviral activity and toxicity in a cervical tissue based organ culture (Project 3, Dr. Gupta), and 4) Evaluation of toxicity and efficacy of CSIC and RC-101 in monkeys (Project 4, Dr. Marx). The scientific studies in these four projects will be supported by the Administrative Core (Core A, Dr. Gupta) and a Formulation Core (Core B, Drs. Rohan and Smith). Dr. Tom Smith (Auritec Pharmaceuticals), serving as the Industry partner in this U19 program, will bring controlled release pellet technologies to develop vaginal ring delivery of RC-101- and CSIC-based microbicides in monkeys. In designing the scope of the proposed topical microbicide program, we have focused on extensively evaluating safety and toxicity of candidate microbicides utilizing in vitro studies, tissue explants and animal models, as well as an expanded and innovative organ culture model to study the antiviral activity of microbicides in the presence of common STIs. Our multidisciplinary approach to microbicide development targets two different sites of HIV-1 replication (entry and reverse transcription) through a controlled-release ring formulated product, utilizes a novel ex vivo organ culture to test toxicity, inflammation and antiviral activity across cervicovaginal mucosa, and applies macaque models to study toxicity and efficacy. Therefore, the proposed study will provide novel strategies to develop microbicides for the prevention of HIV-1 transmission and their sequelae for women.