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.

摘要 HIV-1衣壳蛋白（CA）是开发新型抗逆转录病毒药物的一个有吸引力的靶点，这是由于其 在病毒生物学中的重要和多方面的作用。Lenacapavir（LEN）是最近发现的一流药物， 具有皮摩尔效力的抗HIV-1的长效captain靶向抑制剂。2/3期临床试验 研究表明，皮下注射LEN，间隔6个月给药， 在感染多药耐药病毒表型的重度治疗患者中抑制。这些 研究结果强调LEN是一种有前途的药物，可以补充目前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 鉴定了许多赋予对LEN的实质性抗性的CA置换。具体而言，Q67 H/N74 D和 M66 I取代成为主要的耐药性相关变体。因此，有必要制定 具有更高抗性屏障的改进的LEN类似物。我们在学术上建立了LEN的合成方法 通过利用模块化方法独立地制备三个独特的LEN亚基（A、B和C）， 市售试剂。这种模块化方法允许对每个模块进行简单的修改。 亚基，然后可以以任何顺序组合以制备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。我们互补的病毒学，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CA六聚体的高分辨率X射线晶体结构， 主要耐药相关的Q67 H/N74 D和M66 I变化。我们建议利用这些发现， 合理开发具有更高抗性屏障的改良LEN类似物。在目标1中，我们的努力将集中在 对LEN亚基A和C进行修饰以克服由LEN亚基A和C产生的空间位阻和静电排斥。 耐药Q67 H/N74 D CA变体。在目标2中，我们将使用我们最近从MiniFrags中获得的有希望的发现 筛选研究，其鉴定了结合至与LEN紧密邻近的CA疏水口袋的片段。 我们将通过将这些片段连接到LEN来合成新的化学型，以产生第二代 类似物靶向主要耐药M66 I变体。来自目标1和2的新合成化合物 将通过使用抗病毒活性、细胞毒性、表面等离子体共振和X-射线衍射的逐步方法进行评估。 射线晶体学来鉴定和表征先导化合物。总的来说，拟议的研究是 预期产生对主要耐药CA具有改善的抗病毒活性的新型化学型 赋予对亲本LEN的实质性抗性的变体。