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（由申请人提供）：逆转录病毒 HIV 是全球艾滋病流行的病原体，自 1981 年发现以来一直是一个主要的健康问题。目前，最成功的治疗方案是高效抗逆转录病毒疗法（HAART），该药物结合了目前 FDA 批准的 26 种 HIV 治疗药物中的三到四种，其中一半药物针对病毒复制所必需的逆转录酶 (RT)，其中有 5 种是逆转录酶抑制剂。目前，与 RT 变构口袋结合的非核苷逆转录酶抑制剂 (NNRTI) 可以单独抑制聚合酶功能，或者与与酶活性位点结合的核苷/核苷酸逆转录酶抑制剂 (NRTI) 协同抑制聚合酶功能。尽管 HAART 已成功延长患者的预期寿命，但 NNRTI 始终与 HAART 中的 NRTI 联合使用，以最大限度地发挥协同作用和疗效。 HIV感染者，这种治疗方案存在一些局限性，包括个体药物的溶解度、长期毒副作用和耐药性，因此，这项研究的长期目标是开发更安全、具有改善药理学特性的下一代疗法。 (PK/PD) 和可克服耐药变体的溶解度，我们之前开发了新型 NNRTI，它们对 HIV-1 以及 HIV-1 的耐药变体具有非常有效的抗病毒活性（最佳 EC50 = 530 pM）。此外，我们的化合物比 FDA 批准的第二代 NNRTI 依曲韦林和利匹韦林表现出更低的细胞毒性和更高的溶解度。此外，RT-抑制剂的相互作用将通过X射线晶体学来阐明，从而为合理的药物设计提供平台。此外，新药的抗病毒活性。我们实验室开发的 NNRTI 将使用基于细胞的方法进行测量，并检查与目前 FDA 批准的 NRTI 结合时的效力变化，以找到表现出最高协同效应的组合。最后，研究 NRTI-NNRTI 协同作用的分子机制。将通过解析四元 RT:DNA:NRTI:NNRTI 结构进行研究 总体而言，这项研究将使我们能够开发出更有效、具有改进药理学特性的 NNRTI，并确定 NNRTI 的最佳组合。和 NRTI 联合治疗可降低治疗失败率并延缓耐药性的产生。