Phenotypes of HIV-1 Integrases

描述(申请人提供)：随着对现有抗逆转录病毒药物的耐药性和毒性的增加，必须开发针对人类免疫缺陷病毒(HIV)生命周期中除蛋白酶和逆转录酶以外的各个步骤的抑制剂。整合酶(IN)抑制剂已经进入临床试验。因此，会出现对IN抑制剂的抗药性。具体地说，假设对IN抑制剂的抗药性将对IN的生化和结构以及IN与其抑制剂之间的相互作用产生不利影响。其次，通过合成现有的IN抑制剂衍生的类似物，可以探索IN与其抑制剂之间相互作用的本质。最终，这将导致抑制进入动物和人体试验的整合反应。为了验证这些假设，我们提出了以下具体目标：1.选择IN抑制剂耐药的HIV和MAP耐药突变。2.确定耐药性突变对IN、HIV复制和整合的影响。3.合成DCTA和DKA的类似物，以确定具有更好的抗HIV活性、更好的抗IN活性和更好的细胞进入能力的化合物。4.映射与溶液中的抑制剂结合的氨基酸。这些研究将确定IN对抑制剂产生抗药性的机制，以及这种抗药性对病毒适应性的成本。此外，核磁共振研究将绘制HIV IN蛋白上的抑制剂结合口袋，并确定与IN抑制剂相互作用的特定残基。这些研究的长期目标将是获得对合理合成第二代临床有用的HIV IN抑制剂至关重要的信息，这些抑制剂对IN抑制剂敏感和耐药的HIV分离株都具有活性。一种名为整合酶的HIV蛋白质对HIV复制至关重要，并因此发展为艾滋病。整合酶抑制剂正在进行临床试验。了解艾滋病毒如何对这些化合物产生抗药性以及这些化合物如何与整合酶相互作用是合成更好的抗艾滋病毒药物的基础。

DESCRIPTION (provided by applicant): With increasing resistance and toxicity to existing anti-retroviral agents, it is imperative that inhibitors targeted at steps in the life cycle of the human immunodeficiency virus (HIV) other than protease and reverse transcriptase be developed. Integrase (IN) inhibitors have moved into clinical trials. Therefore, resistance to IN inhibitors will arise. Specifically, it is hypothesized that resistance to IN inhibitors will adversely affect the biochemistry and structure of IN and the interactions between IN and its inhibitors. Second, through synthesis of analogues derived from existing IN inhibitors, the nature of the interactions between IN and its inhibitors can be probed. Ultimately, this will lead to inhibitors of the integration reaction that enter into animal and human testing. To test these hypotheses we propose the following specific aims: 1. Select for IN inhibitor resistant HIV and map resistance mutations. 2. Determine the effects resistance mutations have on IN, HIV replication, and integration. 3. Synthesize analogues of the DCTA's and DKA's to identify compounds with improved anti-HIV activity, improved activity against IN, and improved cellular entry. 4. Map amino acids that bind inhibitors in solution. These studies will determine the mechanisms by which IN becomes resistant to inhibitors and the cost such resistance has on viral fitness. Additionally, the NMR studies will map an inhibitor-binding pocket on the HIV IN protein and identify the specific residues that interact with IN inhibitors. The long-term aim of these studies will be to obtain information critical for the rational synthesis of second generation, clinically-useful inhibitors of HIV IN with activity against both IN inhibitor-sensitive and inhibitor-resistant isolates of HIV. One HIV protein, integrase, is critical for HIV replication and thus progression to AIDS. Inhibitors of integrase are in clinical testing. Understanding how HIV becomes resistant to these compounds and how such compounds interact with integrase are fundamental to the synthesis of better anti-HIV drugs.