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: 10550260
• 负责人: Stefan G Sarafianos
• 金额: $ 47.05万
• 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/10550260
• 关键词: 2' -deoxyadenosine; Acquired Immunodeficiency Syndrome; Action Potentials; 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; 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; Viral; Virus; Work; Zidovudine; analog; antiretroviral therapy; cell immortalization; design; drug resistant virus; experimental study; inhibitor; inhibitor therapy; 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

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.

项目概要 核苷/核苷酸逆转录酶抑制剂 (NRTI) 有效抑制 HIV 并充当骨干 然而，需要新的疗法来持续控制 HIV 感染。 乙炔基-2-氟-2’-脱氧腺苷 (EFdA) 是一种 NRTI，具有卓越的效力、稳定性和独特性 抗 HIV 的作用机制，已获得默克 (Merck) 的许可（也称为 MK-8591 或 Islatavir）。 已成功用于 I 期临床试验，并于近期引入 EFdA 临床试验。 在令人惊讶的低剂量（高达 剂量比现有 NRTI 药物低 10,000 倍以上），旨在每周一次口服和每年一次缓释 因此，这项工作与 NIH 指南直接相关。 高优先级艾滋病资金（NOT-OD-15-137）用于“下一代疗法……效果更长”。 我们已经证明，a) EFdA 在体外、细胞培养物（PBMC 中的 EC50=50 pM）、小鼠和非小鼠中具有高效力。 尽管 EFdA 保留了 3’-OH，但它在人类灵长类动物模型中阻断了 HIV 逆转录酶 (RT)。 体外，主要作为直接/专性，有时由于 EFdA- 的困难而延迟链终止剂 终止病毒DNA易位 因此，EFdA终止了核苷酸逆转录酶易位。 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。 为此，Co-I Malim 建立了一种创新的检测方法，可以对 EFdA 进行研究。 此外，体外传代实验还发现了HIV感染细胞的抗病毒机制。 通过两种范式转变的双重机制或阻力赋予 EFdA 阻力：减少 掺入/增强切除和减少掺入/增强易位 EFdA 类似物的数据。 表明可以有效抑制 EFdA 耐药的 HIV，尽管这种现象的机制尚不清楚 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 高度敏感性，为 EFdA/DOR 联合疗法铺平道路。 EFdA 及其类似物的结构属性影响与不同 RT（野生型、药物- 耐药性，来自不同的进化枝，病毒），导致临床上抑制效率的重要差异， 该假设将通过耐药性、过敏性和生化作用机制进行检验。 三个具体目标，努力结合新颖的测序来回答上述问题， 这项工作将有助于优化联合疗法。 减少药物负担并具有非凡的长效潜力，解决慢性艾滋病毒的依从性挑战 治疗。