DEVELOPMENT OF RETROCYCLIN MICROBICIDES

逆环素类杀菌剂的开发

• 批准号: 6955868
• 负责人: ALEXANDER MICHAEL COLE
• 金额: $ 19.2万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6955868
• 关键词: AIDS education /prevention; antiAIDS agent; bioengineering /biomedical engineering; biological products; chemoprevention; clinical research; defensins; drug design /synthesis /production; drug screening /evaluation; human immunodeficiency virus 1; human subject; local antiinfective agents; mucosa; receptor binding; topical drug application; virus infection mechanism; virus receptors

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.

自行涂抹预防剂来预防粘膜、特别是阴道或直肠传播 HIV-1 的优点是使易受感染的伴侣能够采取有效措施保护自己。在寻找作为杀微生物剂开发的理想模板的分子时，我们利用固相肽合成从其遗骸中重建了一种进化失落的人类抗菌肽“逆环素”，即一种θ防御素假基因。逆周期素有效预防 X4 和 R5 HIV-1 对 CD4+ 细胞感染的能力非常显着。其他研究表明，逆环素的作用模式是抑制 HIV-1 结合和进入的初始步骤。研究发现，下一代逆环素类似物对来自大多数 HIV-1 群体和亚型的大量初级分离株具有活性。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，适合作为局部杀菌剂进行临床前开发。根据我们的研究，我们形成了关于逆周期素的几个假设：A) 逆周期素可能会起到抑制阴道粘膜中 HIV-1 感染的作用，B) 逆周期素和氰基维林 N（另一种有效的抗 HIV-1 凝集素）的糖基化靶点（项目 1) 是不同的，因此它们的活动可能是 互补，C) 逆转录环素是有效的抗逆转录病毒药物，适合进一步开发为局部阴道杀微生物剂，以预防艾滋病毒传播。为了检验这些假设，我们建议：1) 构建并表征下一代抗 HIV 逆环素类似物，2) 评估候选逆环素制剂的抗 HIV-1 活性、稳定性和细胞毒性，以及 3) 评估候选逆环素制剂的抗 HIV-1 活性、稳定性和细胞毒性。人阴道液和血清中的生物功效。虽然断言逆转录素的进化丧失导致人类对 HIV-1 感染的易感性可能是推测性的，但逆转录素是设计可预防 HIV 感染的天然杀微生物剂的有希望的线索。