A Competition Binding Assay for Identifying Novel HIV-1 Capsid Ligands

艾滋病毒/艾滋病仍然是全球流行病和公共卫生威胁。尽管开发了有效的抗逆转录病毒药物，但治疗不能治愈，因此是终身的。长期治疗往往受到病毒耐药性突变和不完全依从性的影响。HIV-1衣壳代表了一个有吸引力的治疗靶点，尽管目前可用的抑制剂缺乏足够的临床开发效力。该项目将开发一种基于竞争的检测方法，用于针对HIV-1衣壳中特定口袋的小分子，并将该检测方法用于大型化合物文库的高通量筛选。该试验将通过筛选1万个小分子库并分析其抗病毒活性和细胞毒性来验证。这项工作将有助于鉴定靶向HIV-1衣壳的抗病毒药物的先导化合物，并将鉴定HIV-1衣壳功能的新探针。具体目标：目标1。建立一种基于竞争的高通量筛选方法，用于检测HIV-1 CA中结合H3-H4口袋的小分子。进行中试筛选以验证分析结果。目标3。测试对已知衣壳抑制剂具有抗性的野生型HIV和突变型病毒的抗病毒活性。

DESCRIPTION: HIV/AIDS remains a global epidemic and public health threat. Despite the development of effective antiretroviral drugs, therapy is not curative and is therefore life-long. Long-term therapy is often compromised by viral drug resistance mutations and incomplete adherence. The HIV-1 capsid represents an attractive target for therapy, though currently available inhibitors lack sufficient potency for clinical development. This project will develop a competition-based assay for small molecules targeting a specific pocket in the HIV-1 capsid and will adapt the assay for high-throughput screening of large compound libraries. The assay will be validated by screening a library of 10,000 small molecules and analyzing the hits for antiviral activity and cytotoxicity. This work will facilitate the identification of lead compounds for development of antivirals targeting the HIV-1 capsid, and will identify novel probes for HIV-1 capsid function. Specific Aims: Aim 1. Establish a competition-based high-throughput screening assay for small molecules that bind the H3-H4 pocket in HIV-1 CA. Aim 2. Perform pilot screens to validate the assay. Aim 3. Test the confirmed hits for antiviral activity against wild type HIV- and mutant viruses that are resistant to known capsid inhibitors.