Development of RC-101 as an Intravaginal Anti-HIV-1 Topical Microbicide

开发 RC-101 作为阴道内抗 HIV-1 局部杀菌剂

• 批准号: 7681867
• 负责人: ALEXANDER MICHAEL COLE
• 金额: $ 40.24万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7681867
• 关键词: Affect; Attention; Binding; Bioavailable; Cells; Cervical; Coitus; Combined Modality Therapy; Complex; Development; Drug Formulations; Epithelial Cells; Epithelium; Erythrocytes; Evaluation; Exhibits; Film; Gel; Goals; HIV; HIV-1; Heterosexuals; Human; Immunologist; In Vitro; Infection; Intravaginal Administration; Knowledge; Liquid substance; Local Microbicides; Macaca nemestrina; Measures; Modeling; Monkeys; Mucous Membrane; Organ Culture Techniques; Peptides; Pharmaceutical Preparations; Physiological; Pilot Projects; Preclinical Testing; Prevention; Principal Investigator; Resistance; Route; Seminal Plasma; Sexual Partners; Sexual Transmission; Silicone Elastomers; Structure; Surface; Testing; Tissues; Toxic effect; U-Series Cooperative Agreements; Vagina; Vaginal Ring; Vaginal Route of Drug Administration; Work; absorption; analog; antimicrobial peptide; base; controlled release; cost; cytokine; cytotoxicity; design; empowered; in vivo; inhibitor/antagonist; interdisciplinary approach; lysine analog; male; microbicide; multidisciplinary; non-nucleoside reverse transcriptase inhibitors; novel; polypeptide; prevent; programs; prophylactic; rectal; retrocyclin; transmission process; vaccine development; vaginal fluid; vaginal microbicide; vaginal transmission

Sexual transmission through mucosal surfaces has been the most common route of HIV-1 spread throughout the world. Although much attention has been focused on vaccine development for HIV-1, progress has been slow and there is an urgent need to find alternative approaches, such as topical microbicides, to target the spread of HIV-1. In our search for novel compounds active against HIV-1, we discovered that the cyclic antimicrobial peptides called retrocyclins were potent inhibitors of HIV-1 replication. We have synthesized and tested the anti-HIV-1 activity of nearly 120 analogs of retrocyclin, and determined that the lysine analog RC-101 was best suited for development as a topical microbicide. RC-101 was shown to be active against R5 and X4 HIV-1, and acts by preventing the initiation of the fusion complex by binding the heptad repeat region 2 on gp41, precluding six helix bundle formation. RC-101 also exhibited little to no cytotoxicity or induction of proinflammatory cytokines in organotypic cervicovaginal epithelia and cervical organ culture, did not hemagglutinate red blood cells, was remarkably stable, and could be formulated into thin films for intravaginal application. Pilot studies with film-formulated RC-101 in five pigtail macaques revealed that the peptide was safe, exhibited no appreciable toxicity, and was associated with cervicovaginal epithelial cells. Based on our findings, we have formed several hypotheses about RC-101: a) RC-101 is stable and bioavailable, thus will likely function to inhibit HIV-1 infection in the vaginal mucosa, b) RC-101 (an entry/fusion inhibitor) and CSIC (a non-nucleoside reverse transcriptase inhibitor being developed by Project 1) have different mechanisms of action and thus their activities will be complementary in preventing HIV transmission, and c) RC-101, formulated as a controlled-release product, can provide long-term protection against heterosexual HIV-1 transmission. With assistance from a multidisciplinary team, we will test these hypotheses by 1) determining the broad spectrum efficacy, stability in biologic fluids and ability to induce HIV-1 resistance of RC-101, alone and in combination with CSIC, and 2) evaluating silicone elastomer ring-formulated RC-101, alone and in combination with CSIC, in organotypic cervicovaginal tissues and following topical vaginal administration in monkeys. These studies involve a highly collaborative and iterative approach to develop RC-101 in combination with CSIC as a ring-formulated intravaginal topical microbicide.

通过粘膜表面的性传播是 HIV-1 传播的最常见途径 全世界。尽管 HIV-1 疫苗的开发受到了很多关注， 进展缓慢，迫切需要寻找替代方法，例如专题 杀微生物剂，以针对 HIV-1 的传播。在寻找具有抗 HIV-1 活性的新型化合物时，我们 发现称为逆转录环素的环状抗菌肽是 HIV-1 的有效抑制剂 复制。我们合成并测试了近120种逆转录素类似物的抗HIV-1活性， 确定赖氨酸类似物 RC-101 最适合开发为局部杀菌剂。 RC-101 被证明对 R5 和 X4 HIV-1 具有活性，并通过阻止融合复合物的启动发挥作用 通过将七肽重复区域 2 结合到 gp41 上，阻止六螺旋束的形成。 RC-101也展出 在器官型宫颈阴道上皮细胞中几乎没有细胞毒性或促炎细胞因子的诱导，并且 宫颈器官培养，红细胞不发生血凝，非常稳定，可以 配制成薄膜供阴道内使用。在五个辫子中使用薄膜配制的 RC-101 进行试点研究 猕猴表明该肽是安全的，没有表现出明显的毒性，并且与 子宫颈阴道上皮细胞。根据我们的发现，我们对 RC-101 提出了几个假设： a) RC-101 稳定且具有生物利用度，因此可能起到抑制阴道粘膜中 HIV-1 感染的作用， b) RC-101（一种进入/融合抑制剂）和CSIC（一种非核苷逆转录酶抑制剂， 项目 1) 开发的产品具有不同的作用机制，因此它们的活动将是互补的 预防 HIV 传播，以及 c) RC-101，配制为控释产品，可以提供 长期防止异性 HIV-1 传播。在多学科的帮助下 团队，我们将通过 1) 确定生物液体中的广谱功效和稳定性来测试这些假设 RC-101 单独或与 CSIC 联合诱导 HIV-1 抗性的能力，以及 2) 评估 有机硅弹性体环配方 RC-101，单独或与 CSIC 组合，有机型 子宫颈阴道组织和猴子局部阴道给药后。这些研究涉及 与 CSIC 联合开发 RC-101 环配方的高度协作和迭代方法 阴道内局部杀菌剂。