Development of Improved HIV-1 Capsid Inhibitors

改进的 HIV-1 衣壳抑制剂的开发

• 批准号: 10700506
• 负责人: Daniel Adu-Ampratwum
• 金额: $ 24.99万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-04 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10700506
• 关键词: Academia; Anti-Retroviral Agents; Antiviral Agents; Binding; Biochemical; Biochemistry; Biological Assay; Biology; Capsid; Capsid Proteins; Cell Culture Techniques; Cells; Chemical Structure; Clinical Trials; Complement; Coupled; Data; Development; Dose; Drug Industry; Drug Targeting; Drug resistance; Electrostatics; Generations; Goals; HIV; HIV resistance; HIV-1; HIV-1 drug resistance; Hydrophobicity; Infection; Investigation; Lead; Life Cycle Stages; Modification; Multi-Drug Resistance; Patients; Persons; Pharmaceutical Chemistry; Phase II/III Clinical Trial; Phenotype; Process; Proteins; Publishing; Reagent; Research; Resistance; Resistance profile; Resolution; Roentgen Rays; Role; Scanning; Science; Structure; Structure-Activity Relationship; Surface Plasmon Resonance; Testing; Variant; Viral; Viral Physiology; Virus; Virus Replication; X-Ray Crystallography; analog; base; cytotoxicity; design; drug resistant virus; experience; experimental study; follow-up; functional group; gag Gene Products; improved; inhibitor; multimodality; novel; prevent; rational design; screening; small molecule; structural biology; subcutaneous; virology

ABSTRACT The HIV-1 capsid protein (CA) is an attractive target for the development of novel antiretrovirals due to its essential and multifaceted roles in the virus biology. Lenacapavir (LEN) is a recently discovered first-in-class, long-acting capsid-targeting inhibitor with picomolar potency against HIV-1. Phase 2/3 clinical trials have revealed that subcutaneous administration of LEN with a six-month dosing interval enables high rates of HIV-1 suppression in heavily treatment-experienced patients infected with multi-drug resistant viral phenotypes. These findings have highlighted LEN as a promising agent that could complement current antiretroviral compounds to treat people living with HIV-1. However, cell culture-based viral breakthrough assays and clinical trials have identified a number of CA substitutions that confer substantial resistance to LEN. Specifically, Q67H/N74D and M66I substitutions emerged as major drug-resistance associated variants. Therefore, there is a need to develop improved LEN analogs with a higher barrier to resistance. We have established synthesis of LEN in academic setting by utilizing a modular approach of independently preparing three unique LEN subunits (A, B, and C) from commercially available reagents. Such modular approach allows for straightforward modifications of each subunit which can then be combined in any order to prepare LEN and its analogs. Thus, our medicinal chemistry strategy is highly advantageous for analog development. Our complementary virology, biochemistry and structural biology experiments have allowed us to characterize a multi-modal mechanism of action of LEN. Furthermore, we have recently determined high-resolution X-ray crystal structures of CA hexamers containing major drug-resistance associated Q67H/N74D and M66I changes. We propose to exploit these findings to rationally develop improved LEN analogs with a higher barrier to resistance. In Aim 1, our efforts will focus on modifications to LEN subunits A and C to overcome steric hindrance and electrostatic repulsions created by the drug resistant Q67H/N74D CA variant. In aim 2, we will use our recent promising findings from MiniFrags screening studies, which identified fragments that bind to the CA hydrophobic pocket in close vicinity to LEN. We will synthesize new chemotypes by connecting these fragments to LEN to generate second-generation analogs to target the major drug-resistant M66I variant. Newly synthesized compounds from both Aims 1 and 2 will be evaluated by a stepwise approach using antiviral activity, cytotoxicity, surface plasmon resonance and X- ray crystallography to identify and characterize the lead compound(s). Taken together, the proposed research is expected to generate novel chemotypes with improved antiviral activities against major drug resistant CA variants that confer substantial resistance to parental LEN.

抽象的 HIV-1 衣壳蛋白 (CA) 是新型抗逆转录病毒药物开发的一个有吸引力的靶标，因为它具有 在病毒生物学中发挥重要和多方面的作用。 Lenacapavir (LEN) 是最近发现的一种一流的、 长效衣壳靶向抑制剂，具有皮摩尔抗 HIV-1 效力。 2/3期临床试验已 研究表明，以六个月的给药间隔皮下注射 LEN 可提高 HIV-1 感染率 对感染多重耐药病毒表型的经过大量治疗的患者进行抑制。这些 研究结果强调 LEN 是一种有前途的药物，可以补充现有的抗逆转录病毒化合物 治疗 HIV-1 感染者。然而，基于细胞培养的病毒突破性测定和临床试验已经 确定了许多 CA 替代，这些替代对 LEN 具有显着的抗性。具体来说，Q67H/N74D 和 M66I 取代成为主要的耐药相关变异。因此，需要开发 改进的 LEN 类似物具有更高的耐药屏障。我们已经在学术上建立了 LEN 的综合 通过利用模块化方法独立制备三个独特的 LEN 亚基（A、B 和 C）来进行设置 市售试剂。这种模块化方法允许直接修改每个 然后可以以任何顺序组合亚基以制备 LEN 及其类似物。因此，我们的药物化学 策略对于模拟开发非常有利。我们的互补病毒学、生物化学和 结构生物学实验使我们能够表征 LEN 的多模式作用机制。 此外，我们最近确定了包含 CA 六聚体的高分辨率 X 射线晶体结构 主要耐药性相关的 Q67H/N74D 和 M66I 变化。我们建议利用这些发现 合理开发具有更高耐药屏障的改良LEN类似物。在目标 1 中，我们的努力将集中于 对 LEN 亚基 A 和 C 进行修饰，以克服由 耐药 Q67H/N74D CA 变体。在目标 2 中，我们将利用 MiniFrags 最近有希望的发现 筛选研究，鉴定了与 LEN 附近的 CA 疏水口袋结合的片段。 我们将通过将这些片段连接到 LEN 来合成新的化学型以生成第二代 类似物以针对主要耐药性 M66I 变体。来自目标 1 和 2 的新合成化合物 将使用抗病毒活性、细胞毒性、表面等离振子共振和X-通过逐步方法进行评估 射线晶体学来识别和表征先导化合物。总而言之，拟议的研究是 预计将产生新的化学型，对主要耐药 CA 具有更好的抗病毒活性 赋予亲本 LEN 显着抗性的变异体。