Optimizing HIV reverse transcriptase inhibitors through structural, kinetic and cellular approaches

通过结构、动力学和细胞方法优化 HIV 逆转录酶抑制剂

• 批准号: 9378691
• 负责人: Albert Hay Wah Chan
• 金额: $ 5.71万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9378691
• 关键词: Acquired Immunodeficiency Syndrome; Active Sites; Address; Adverse effects; Affinity; Anti-HIV Agents; Antiviral Agents; Binding; Biological Assay; Biology; Catechols; Cells; Combined Modality Therapy; Complex; Computing Methodologies; Crystallization; DNA; Development; Dissociation; Docking; Drug Design; Drug Targeting; Drug resistance; Enzyme Inhibitor Drugs; Enzymes; Epidemic; Etiology; Exhibits; FDA approved; Free Energy; Future; Generations; Goals; HIV; HIV therapy; HIV-1; Health; Highly Active Antiretroviral Therapy; Human; Individual; Kinetics; Knowledge; Life Expectancy; Literature; Measures; Methods; Modeling; Molecular; Mutation; Nucleosides; Nucleotides; Pathogenesis; Pharmaceutical Preparations; Pharmacology; Polymerase; Process; Property; RNA-Directed DNA Polymerase; Research; Resistance; Resistance development; Resistance profile; Retroviridae; Reverse Transcriptase Inhibitors; Safety; Series; Solubility; Structure; Structure-Activity Relationship; System; T-Lymphocyte; Testing; Tetrazolium; Therapeutic; Toxic effect; Treatment Failure; Treatment Protocols; Uracil; Variant; Viral Load result; Virus; Virus Replication; Work; X-Ray Crystallography; base; cytotoxicity; design; improved; inhibitor/antagonist; non-nucleoside reverse transcriptase inhibitors; novel; novel therapeutics; public health relevance; structural biology; synergism; transmission process

 DESCRIPTION (provided by applicant): Retrovirus HIV, the etiological agent of the global AIDS epidemic, has been a major health concern since its discovery in 1981. Currently, the most successful treatment regimen is the highly active anti-retroviral therapy (HAART), which combines three or four of the current 26 FDA-approved HIV drugs in treatment. Half of these drugs target the reverse transcriptase (RT) enzyme that is essential for viral replication. Among the RT inhibitors, five are non-nucleoside reverse transcriptase inhibitors (NNRTIs) which bind to an allosteric pocket of RT. NNRTIs can inhibit the polymerase function by themselves, or synergistically with nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs) that bind to the active site of the enzyme. Currently, NNRTIs are always combined with NRTIs in HAART to maximize synergy and efficacy. Although HAART has been successful in extending the life expectancy of HIV-infected individuals, several limitations to this treatment regimen exist, including solubility of individual drug, long-term toxic side effects and drug resistance. Therefore, the long term goal of this research is to develop safer, next generation therapeutics with improved pharmacological properties (PK/PD) and solubility that can overcome resistant variants. Using computational approaches, we have previously developed novel NNRTIs that have very potent antiviral activity (best EC50 = 530 pM) against HIV-1 as well as resistant variants of HIV-1 with common RT mutations. Moreover, our compounds exhibit much lower cytotoxicity and higher solubility than the FDA-approved second generation NNRTIs etravirine and rilpivirine. In order to further develop these compounds into useful therapeutics, binding affinity and association/dissociation rates will be determined using pre-steady state kinetics. Moreover, RT- inhibitor interactions will be elucidated with x-ray crystallography to provide a platform for rational drug design. Furthermore, antiviral activity of novel NNRTIs developed in our lab will be measured using cell-based assays, and changes in potency when combined with current FDA-approved NRTIs will be examined to find the combination that displays the highest synergistic effect. Finally, the molecular mechanism of NRTI-NNRTI synergy will be investigated by solving a quaternary RT:DNA:NRTI:NNRTI structure. Overall, this research will allow us to develop more potent NNRTIs with improved pharmacological properties, and identify the optimal combination of NNRTI and NRTI for combination therapy that should lower treatment failure rate and delay development of resistance.

 描述（由适用提供）：自1981年发现以来，全球艾滋病流行病的病因逆转录病毒HIV一直是一个主要的健康问题。目前，最成功的治疗方案是高度活跃的抗逆转录病毒疗法（HAART），它结合了当前26 FDA批准的HIV HIV药物中的三个或四个治疗中的三个或四个。这些药物的一半靶向逆转录酶（RT）酶，这对于病毒复制至关重要。在RT抑制剂中，有5个是非核苷逆转录酶抑制剂（NNRTIS），它们与RT的变构袋结合。 NNRTI可以自身抑制聚合酶功能，或与核苷/核苷逆转录酶抑制剂（NRTI）合成，该酶与酶的活性位点结合。目前，NNRTI在HAART中总是与NRTI结合起来，以最大程度地提高协同作用，尽管Haart成功地延长了感染了HIV感染的个体的预期寿命，但存在这种治疗方案的局限性，包括个人药物的解决方案，长期毒性副作用和耐药性。因此，这项研究的长期目标是开发具有改进的药物特性（PK/PD）和可溶性的更安全，下一代疗法，可以克服抗性变体。使用计算方法，我们以前已经开发了新型的NNRTI，它们具有非常潜在的抗病毒活性（最佳EC50 = 530 pm），以及HIV-1和HIV-1具有常见RT突变的抗性变体。此外，与FDA批准的第二代Nnrtis etravirine和rilpivirine相比，我们的化合物表现出低得多的细胞毒性和更高的溶解度。为了将这些化合物进一步发展为有用的治疗，将使用稳态的状态动力学确定结合亲和力/解离率。此外，将使用X射线晶体学阐明RT抑制剂相互作用，以提供一个理性药物设计的平台。此外，将使用基于细胞的测定法测量我们实验室中开发的新型NNRTI的抗病毒活性，并且将检查与当前FDA批准的NRTI相结合时的效力变化，以找到显示最高的组合。协同效应。最后，通过求解第四纪RT：DNA：NRTI：NNRTI结构，将投资NRTI-NNRTI协同作用的分子机制。总体而言，这项研究将使我们能够通过改进的药物特性开发更多的潜在nnrtis，并确定NNRTI和NRTI的最佳组合用于组合疗​​法，以降低治疗失败率并延迟抗性的发展。