DEVELOPMENT OF RETROCYCLIN MICROBICIDES

自我施用预防剂以防止HIV-1的粘膜传播，特别是阴道或直肠传播，其优点是使易受感染的伴侣能够采取有效措施保护自己。在我们的分子，这将是理想的杀微生物剂开发的模板搜索，我们利用固相肽合成重建一个进化丢失的人类抗菌肽“retrocyclin”从它的遗体，一个θ-防御素假基因。逆转录酶的能力，有效地防止感染的CD 4+细胞的X4和R5 HIV-1是显着的。进一步的研究表明，逆转录酶抑制HIV-1结合和进入的初始步骤作为其作用模式。下一代逆转录酶类似物被发现对来自大多数HIV-1组和亚型的许多主要分离株具有活性。重要的是，我们已经确定了一种先导化合物RC-101，它对HIV-1具有高度活性，无细胞毒性，适合作为局部杀微生物剂进行临床前开发。基于我们的研究，我们已经形成了几个关于逆转录细胞周期素的假设：A）逆转录细胞周期素可能具有抑制阴道粘膜中HIV-1感染的功能，B）逆转录细胞周期素的糖基化靶点和另一种有效的抗HIV-1凝集素cyanovirin N（项目1）是不同的，因此它们的活性可能不同。 互补的，和C）逆转录病毒素是有效的抗逆转录病毒剂，适合进一步开发作为局部阴道杀微生物剂以预防HIV传播。为了验证这些假设，我们建议：1）构建和表征下一代抗HIV逆转录病毒类似物，2）评价候选逆转录病毒制剂的抗HIV-1活性、稳定性和细胞毒性，3）评价候选逆转录病毒制剂在人阴道液和血清中的生物学功效。虽然它可能是推测性的断言，逆转录病毒的进化损失有助于人类对HIV-1感染的易感性，逆转录病毒是有希望的领导设计天然存在的杀微生物剂，可以预防HIV感染。

Self-applied prophylactic agents to prevent mucosal, particularly vaginal or rectal, transmission of HIV-1 have the advantage of empowering vulnerable receptive partners to take effective measures for their own protection. In our search for molecules that would be ideal templates for microbicide development, we utilized solid-phase peptide synthesis to recreate an evolutionary lost human antimicrobial peptide "retrocyclin" from its remains, a theta-defensin pseudogene. The ability of retrocyclin to potently prevent infection of CD4+ cells by both X4 and R5 HIV-1 was remarkable. Additional studies revealed that retrocyclin inhibits the initial steps in HIV-1 binding and entry as its mode of action. Next-generation analogs of retrocyclin were found to be active against numerous primary isolates from most groups and subtypes of HIV-1. Importantly, we have identified a lead compound, RC-101, that is highly active against HIV-1, non-cytotoxic, and suitable for preclinical development as a topical microbicide. Based on our studies, we have formed several hypotheses about retrocyclins: A) retrocyclins will likely function to inhibit HIV-1 infection in the vaginal mucosa, B) the glycosylated targets of retrocyclins and cyanovirin N, another potent anti-HIV-1 lectin (Project 1), are different and thus their activities may be complementary, and C) retrocyclins are potent antiretroviral agents suitable for further development as topical vaginal microbicides to prevent HIV transmission. To test these hypotheses, we propose to: 1) Construct and characterize next-generation analogs of anti-HIV retrocyclins, 2) Evaluate candidate retrocyclin formulations for anti-HIV-1 activity, stability and cytotoxicity, and 3) Evaluate candidate retrocyclin formulations for biologic efficacy in human vaginal fluid and serum. While it may be speculative to assert that the evolutionary loss of retrocyclin contributed to the susceptibility of humans to HIV-1 infection, retrocyclins are promising leads for designing naturally occuring microbicides that can prevent HIV infections.