Ultrapotent Inhibitors of Wild-type and Multi-drug Resistant HIV

野生型和多重耐药艾滋病毒的超强抑制剂

• 批准号: 9927260
• 负责人: Stefan G Sarafianos
• 金额: $ 49.88万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9927260
• 关键词: 2' -deoxyadenosine; Acquired Immunodeficiency Syndrome; Address; Adherence; Affect; Animal Model; Anti-HIV Agents; Antiviral Agents; Belief; Binding; Binding Sites; Biochemical; Biological Assay; Cell Culture Techniques; Cell Line; Cells; Chronic; Clinic; Clinical; Combined Modality Therapy; Complex; Crystallization; Data; Development; Dose; Drug Combinations; Drug resistance; Event; Excision; Funding; Generations; Genomics; Guidelines; HIV; HIV Infections; HIV drug resistance; HIV resistance; HIV therapy; In Vitro; Licensing; Modality; Multi-Drug Resistance; Mus; Mutation; Names; Nucleosides; Nucleotides; Oral; Outcome; Paper; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase; Phase II Clinical Trials; Prevention; Publishing; RNA-Directed DNA Polymerase; Research; Resistance; Resistance profile; Reverse Transcriptase Inhibitors; Reverse Transcription; Site; Structure; System; Tenofovir; Testing; Time; United States National Institutes of Health; Vertebral column; Virus; Work; Zidovudine; analog; antiretroviral therapy; base; cell immortalization; design; drug resistant virus; experimental study; inhibitor/antagonist; innovation; insight; mutant; next generation; next generation sequencing; non-nucleoside reverse transcriptase inhibitors; nonhuman primate; novel; novel therapeutics; nucleoside analog; optimal treatments; preclinical development; reconstitution; resistance mutation; viral DNA; virology

PROJECT SUMMARY Nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs) efficiently suppress HIV and serve as backbone of antiretroviral therapies. However, new therapeutics are needed for continued control of HIV infection. 4’- Ethynyl-2-fluoro-2’-deoxyadenosine (EFdA) is an NRTI with exceptional potency, stability, and unique mechanism of action against HIV, which has been licensed by Merck. EFdA (also known as MK-8591 or Islatavir) has been successfully used in Phase I and recently introduced in Phase II clinical trials. EFdA has generated “compelling early results for both treatment and prevention” in patients, tested at remarkably low doses (up to >10,000-fold lower doses than current NRTI drugs) aiming at once-weekly oral and once-yearly slow-release dosing, unprecedented modalities for HIV therapies. Hence, this work is directly relevant to NIH guidelines for high priority AIDS funding (NOT-OD-15-137) for “next generation therapies…that are longer acting.” We have shown that a) EFdA has high potency in vitro, in cell culture (EC50=50 pM in PBMCs), mice and non- human primate animal models. Although EFdA retains the 3’-OH, it blocks HIV reverse transcriptase (RT) in vitro, primarily as an immediate/obligate and at times delayed chain terminator due to difficulty of EFdA- terminated viral DNA to translocate. Thus, EFdA is termed a nucleotide reverse transcriptase translocation inhibitor (NRTTI). However, the inhibition mechanism of EFdA in HIV-infected cells (primary or cell lines) remains unknown. Toward that end, Co-I Malim has established an innovative assay that enables study of the EFdA antiviral mechanism in HIV-infected cells. Additionally, in vitro passage experiments have identified mutations that impart EFdA resistance through two paradigm-shifting dual mechanisms or resistance: decreased incorporation/enhanced excision and decreased incorporation/enhanced translocation. Data on an EFdA analog suggest efficient inhibition of EFdA-resistant HIV, although the mechanism of this phenomenon is not understood. Notably, treatment with key new generation NNRTI doravirine (DOR) leads to resistance mutations that impart hypersusceptibility to EFdA, paving the way for EFdA/DOR combination therapies. We hypothesize that the structural attributes of EFdA and its analogs impact interactions with diverse RTs (wild-type, drug- resistant, from diverse clades, viruses), leading to clinically important differences in efficiency of inhibition, resistance, hypersusceptibility, and biochemical mechanism of action. This hypothesis will be tested through three specific aims, which endeavor to answer the above questions using a combination of novel sequencing, biochemical, structural, and virological approaches. This work will help optimize combination therapies that reduce drug burden and have exceptional long-acting potential, addressing adherence challenges of chronic HIV treatment.

项目摘要 核苷/核苷酸逆转录酶抑制剂（NRTI）有效抑制HIV，并作为骨架 抗逆转录病毒疗法。然而，需要新的治疗方法来持续控制HIV感染。4'- 乙炔基-2-氟-2 '-脱氧腺苷（EFdA）是一种NRTI，具有卓越的效力，稳定性和独特的 抗艾滋病毒的作用机制，已获得默克公司的许可。EFdA（也称为MK-8591或Islatavir） 已成功用于I期临床试验，并于近期引入II期临床试验。EFdA已生成 在患者中进行的“治疗和预防的令人信服的早期结果”，在非常低的剂量下进行测试（高达 剂量比目前的NRTI药物低10，000倍），旨在每周一次口服和每年一次缓释 剂量，前所未有的艾滋病毒治疗方式。因此，这项工作与NIH指南直接相关， 高优先艾滋病资金（NOT-OD-15-137）用于“下一代长效疗法.”。 我们已经表明，a）EFdA在体外、在细胞培养物（在PBMC中EC 50 =50 pM）、小鼠和非哺乳动物中具有高效力， 人类灵长类动物模型。虽然EFdA保留了3 '-OH，但它在体内阻断HIV逆转录酶（RT）。 体外，主要作为立即/专性链终止剂，有时由于EFdA- 终止病毒DNA的转移因此，EFdA被称为核苷酸逆转录酶易位 抑制剂（NRTTI）。然而，EFdA在HIV感染细胞（原代或细胞系）中的抑制机制仍然存在 未知为此，Co-I Malim建立了一种创新的检测方法，可以研究EFdA HIV感染细胞的抗病毒机制。此外，体外传代实验已经鉴定出突变 通过两种范式转换双重机制或抗性赋予EFdA抗性：降低 掺入/增强的切除和降低的掺入/增强的易位。EFdA类似物的数据 表明有效抑制EFdA耐药HIV，尽管这种现象的机制不是 明白值得注意的是，使用关键的新一代NNRTI多拉韦林（DOR）治疗会导致耐药突变 赋予EFdA过敏性，为EFdA/DOR联合治疗铺平了道路。我们假设 EFdA及其类似物的结构属性影响与不同RT（野生型、药物- 抗性，来自不同的进化枝，病毒），导致抑制效率的临床重要差异， 抗性、过敏性和生化作用机制。这一假设将通过 三个具体的目标，奋进用新的测序， 生物化学、结构和病毒学方法。这项工作将有助于优化组合疗法， 减少药物负担，并具有特殊的长效潜力，解决慢性艾滋病毒的依从性挑战 治疗