Discovering Novel Inhibitors of HIV-1 Maturation

描述(申请人提供)：尽管有有效的艾滋病毒抗病毒疗法可用，但仍然需要确定和开发抑制人类免疫缺陷病毒复制的新药，人类免疫缺陷病毒是艾滋病的病原体。本实验室和其他实验室最近的研究对小分子成熟剂3-O-{3‘，3’-二甲基丁二酰基}-白桦酸(DSB)的作用机理进行了表征。DSB是一种有效的、高选择性的HIV-1复制抑制剂，它通过特异性干扰病毒成熟过程中GAG中CA-SP1位点的切割来发挥作用，导致不完全成熟的颗粒，感染性差。我们最近的研究表明，DSB与未成熟的HIV-1颗粒特异结合，这种结合与HIV-1对DSB抗病毒活性的敏感性有关。CA-SP1裂解位点是HIV-1对小分子抑制剂的脆弱性，再加上抗病毒活性与DSB结合有关的观察结果，表明可以通过筛选竞争性抑制DSB与未成熟HIV-1颗粒结合的化合物来识别更多抑制HIV-1成熟的分子。在R21的应用中，我们建议利用我们在HIV-1分子生物学方面的专业知识和我们对DSB抗病毒机制的新理解，开发一种基于SPA的DSB与未成熟HIV-1假病毒粒子结合的快速检测方法。一旦建立了检测方法，我们将进一步优化和调整该系统，应用于大型化合物文库的高通量分子筛选，以确定抑制这种相互作用的分子，作为开发新型HIV-1成熟抑制剂的潜在线索。

DESCRIPTION (provided by applicant): Despite the availability of potent HIV antiviral therapies, there exists a continuing need to identify and develop new drugs that inhibit the replication of the human immunodeficiency viruses, the causative agents of AIDS. Recent studies in our laboratory and others have characterized the mechanism of action of the small molecule maturation inhibitor 3-O-{3',3'-dimethylsuccinyl}-betulinic acid (DSB). DSB is a potent and highly selective inhibitor of HIV-1 replication that acts by specifically interfering with cleavage of the CA-SP1 site in Gag during viral maturation, resulting in incompletely matured particles that are poorly infectious. Our recent studies have shown that DSB binds specifically to immature HIV-1 particles, and this binding was correlated with HIV-1 sensitivity to DSB antiviral activity. The identification of the CA-SP1 cleavage site as an HIV-1 vulnerability to a small molecule inhibitor, coupled with the observation that antiviral activity is linked to DSB binding, suggests that additional molecules that inhibit HIV-1 maturation may be identified by screening for compounds that competitively inhibit DSB binding to immature HIV-1 particles. In this R21 application, we propose to capitalize our expertise in HIV-1 molecular biology and our new understanding of the DSB antiviral mechanism to develop a rapid SPA-based assay for the binding of DSB to immature HIV-1 pseudovirions. Once the assay has been established, we will further optimize and adapt the system for application to high throughput molecular screening of large compound libraries to identify molecules that inhibit the interaction as potential leads for the development of novel HIV-1 maturation inhibitors.