HIV-1 RT dimerization as an antiviral target

描述（由申请人提供）：组合抗逆转录病毒疗法在控制AIDS的进展和延长HIV感染患者的生命方面非常有效。然而，虽然目前的药物治疗可以延缓疾病的进展，但它不能根除病毒，而且它的使用很容易导致抗药性艾滋病毒株的出现。此外，病毒对一种药物的耐药性的出现经常导致对表现出类似化学结构和作用机制的其他药物的交叉耐药性。因此，识别新的病毒靶标和/或开发新类别的抗病毒化合物在对抗HIV/AIDS中是必不可少的。 HIV-1逆转录酶（RT）是一种异二聚体酶，由66 kDa亚基（p66）和p66衍生的51 kDa亚基（p51）组成。该酶的DNA聚合酶和核糖核酸酶H（RNase H）活性完全依赖于该酶的二聚体结构。因此，HIV-1 RT的二聚化为鉴定一类新的抗病毒化合物提供了新的治疗靶点。我们最近开发了一种体外高通量筛选（HTS）试验，可重复检测HIV-1 RT的p66和p51亚基之间的亚基间相互作用。此外，该试验对以前已被证明抑制或增强酶的亚基间相互作用的化合物敏感。鉴于此，本提案中描述的项目包括两个特定目的：（1）优化和验证HIV-1 RT二聚化的体外HTS试验;（2）筛选HIV-1 RT二聚化抑制剂的化学和天然产物文库。 由于其在HIV-1复制中的重要作用，RT已经成为开发抗病毒化合物的主要目标。然而，HIV-1 RT二聚化抑制剂的阐明和/或鉴定将产生一种新的治疗类药物，其表现出与目前用于治疗HIV-1感染个体的核苷和非核苷RT抑制剂完全不同的作用机制。

DESCRIPTION (provided by applicant): Combinatorial anti-retroviral therapies have been remarkably effective in controlling the progression of AIDS and in prolonging the life of patients infected by HIV. However, while the current drug therapy can delay the progression of the disease it can not eradicate the virus, and its use readily leads to the emergence of drug resistant HIV strains. Furthermore, the emergence of viral resistance to one drug frequently results in a cross resistance to other drugs that exhibit similar chemical structures and mechanisms of action. Thus, the identification of novel viral targets and/or the development of new classes of antiviral compounds are essential in the fight against HIV/AIDS. HIV-1 reverse transcriptase (RT) is a heterodimeric enzyme consisting of a 66-kDa subunit (p66) and a p66- derived 51-kDa subunit (p51). The DNA polymerase and ribonuclease H (RNase H) activities of the enzyme are entirely dependent on the dimeric structure of the enzyme. Accordingly, dimerization of HIV-1 RT provides a novel therapeutic target for the identification of a new class of antiviral compounds. We have recently developed an in vitro high throughput screening (HTS) assay that reproducibly detects the intersubunit interactions between the p66 and p51 subunits of HIV-1 RT. Furthermore this assay is sensitive to compounds that have previously been shown to either inhibit or enhance the inter-subunit interactions of the enzyme. In light of this, the project described in this proposal comprises two Specific Aims: (1) to optimize and validate the in vitro HTS assay for HIV-1 RT dimerization; and (2) to screen a chemical and a natural product library for inhibitors of HIV-1 RT dimerization. Due to its essential role in HIV-1 replication, RT is already a primary target for the development of antiviral compounds. However, the elucidation and/or identification of inhibitors of HIV-1 RT dimerization would create a new therapeutic class of drugs that would exhibit mechanisms of action entirely different from the nucleoside and nonnucleoside RT inhibitors that are currently used in the treatment of HIV-1 infected individuals.