Investigation of ATV-Based Heterobifunctional Degraders to Combat Growing HIV-1 PR Inhibitor Resistance

研究基于 ATV 的异双功能降解剂以对抗日益增长的 HIV-1 PR 抑制剂耐药性

基本信息

批准号：
10676954
负责人：
CHRISTINA OCHSENBAUER
金额：
$ 19.64万
依托单位：
UNIVERSITY OF ALABAMA IN TUSCALOOSA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-04 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10676954
关键词：
Affinity Antiviral Agents Atazanavir Autoimmune Diseases Binding Binding Proteins Binding Sites Biological Biological Assay Biological Availability Biology of HIV Infection Cell membrane Cells Clinic Clinical Trials Complex Computing Methodologies Development Dimerization Drug resistance Epidemic Event Exhibits Failure Foundations Future Goals HIV HIV Infections HIV Protease HIV Protease Inhibitors HIV resistance HIV-1 HIV-1 protease Impairment Infection Investigation Length Ligands Ligase Mediating Membrane Modality Modeling Molecular Conformation Multi-Drug Resistance Mutation Nuclear Organic Synthesis Patients Peptide Hydrolases Pharmaceutical Chemistry Pharmaceutical Preparations Predisposition Probability Process Property Protease Inhibitor Proteins Reagent Regimen Resistance Resistance development Series Site Structure Testing Therapeutic Therapeutic Intervention Ubiquitination Variant Viral Viral Physiology Virion Visit Work analog antimicrobial drug antiretroviral therapy assay development combat design dimer drug development drug discovery experience high reward high risk in vitro activity in vivo inhibitor lead optimization member membrane assembly next generation novel novel strategies novel therapeutic intervention particle pol Gene Products pre-exposure prophylaxis prevent prospective prototype recruit refractory cancer resistant strain screening simulation small molecule inhibitor trend ubiquitin-protein ligase viral resistance

项目摘要

PROJECT SUMMARY/ABSTRACT Heterobifunctional targeted protein degraders (TPDs) offer advantages over traditional occupancy-based inhibitors including a unique catalytic mechanism of action (MOA), greater target selectivity, and a reduced probability for resistance development. TPDs are known to effectively target not only cytosolic proteins, but also nuclear and membrane-bound proteins. This therapeutic modality shows great promise for treating drug-resistant cancers and autoimmune diseases but has seen only limited application in antiviral drug discovery. HIV-1 protease (PR) is essential for proteolytic cleavage of Gag and Gag-Pol polyproteins and, thus, virion maturation and infectivity. Gag-Pol forms dynamic dimers at plasma membrane assembly/budding sites, allowing the embedded precursor PR to dimerize as a prerequisite for auto-processing. We identified precursor PR/Gag-Pol as a promising target for protease inhibitor (PI)-based TPDs. The widely used HIV-1 PI, Atazanavir (ATV), is amenable to conjugation with linkers and ubiquitin E3 ligase recruiting ligands to serve as prototype HIV-1 TPDs. Importantly, ATV inhibits activity of mature PR as well as autoprocessing of precursor PR/Gag-Pol during the assembly and budding processes. The OBJECTIVE of this study is to show proof-of-concept that HIV-1 Gag- Pol assembling on the inner leaflet of the plasma membrane can be targeted for aberrant ubiquitination, degradation, and/or inhibition, thereby impairing HIV infectivity. Importantly, due to TPDs’ established MOA, even low-affinity Gag-Pol/TPD interactions are likely to lead to impaired Gag-Pol function. Thus, we pose the HYPOTHESIS that novel ATV-based TPDs will not only augment the inhibition of ATV-sensitive HIV-1 strains compared to ATV, but also exhibit increased and broader biological activity against drug-resistant variants. The objective of AIM 1 is to design and synthesize ATV-based TPDs built on state-of-the-art computational methods and predictive physicochemical properties currently accepted for in vivo active TPDs. Our approach will feature a modular TPD design to establish the appropriate ATV attachment point, E3 ligase recruiter, and length and composition of linker required for HIV-1 Gag-Pol ubiquitination/proteasomal degradation. The goal of AIM 2 is to provide proof-of-concept that ATV-TPDs exert superior activity versus ATV against wild type and PI-resistant HIV-1 strains. We will systematically screen four series of novel ATV-TPD for activity in vitro, in single-round infectivity, and in multi-round replication assays to identify the most promising ATV-TPD candidates. The latter will be tested in complementary limited-scope mechanistic studies, including comparing ATV-TPDs with analogs bearing inactive E3-ligase ligands, to probe if antiviral activity is consistent with the proposed MOA. The IMPACT will be TPDs targeting HIV-1 Gag-Pol and precursor PR that limit infectivity and replication through a mechanism distinct from occupancy-based HIV-1 PI. This will spur the future development of efficacious regimens against PI-resistant HIV strains with reduced susceptibility for resistance development.

项目概要/摘要异双功能靶向蛋白质降解剂 (TPD) 比传统的基于占用的蛋白质降解剂具有优势抑制剂包括独特的催化作用机制 (MOA)、更高的靶标选择性以及更低的众所周知，TPD 不仅能有效靶向胞质蛋白，还能有效靶向胞质蛋白。这种治疗方式在治疗耐药性方面显示出巨大的前景。癌症和自身免疫性疾病，但在抗病毒药物发现中的应用有限。蛋白酶 (PR) 对于 Gag 和 Gag-Pol 多蛋白的蛋白水解切割以及病毒颗粒的成熟至关重要 Gag-Pol 在质膜组装/出芽位点形成动态二聚体，从而允许嵌入前体 PR 进行二聚化是自动处理的先决条件，我们确定了前体 PR/Gag-Pol。广泛使用的 HIV-1 PI 阿扎那韦 (ATV) 是基于蛋白酶抑制剂 (PI) 的 TPD 的一个有前景的靶标。适合与接头和泛素 E3 连接酶招募配体缀合，作为原型 HIV-1 TPD。重要的是，ATV 抑制成熟 PR 的活性以及前体 PR/Gag-Pol 的自动加工过程。本研究的目的是证明 HIV-1 Gag- 的概念验证。质膜内叶上的 Pol 组装可针对异常泛素化，重要的是，这会损害 HIV 的传染性，甚至影响 TPD 已建立的 MOA。低亲和力的 Gag-Pol/TPD 相互作用可能会导致 Gag-Pol 功能受损。假设新型基于 ATV 的 TPD 不仅会增强对 ATV 敏感的 HIV-1 毒株的抑制作用与 ATV 相比，它还表现出针对耐药变异体的更高和更广泛的生物活性。 AIM 1 的目标是设计和综合基于 ATV 的 TPD，该 TPD 建立在最先进的计算基础上我们的方法和目前接受的体内活性 TPD 的预测物理化学特性。将采用模块化 TPD 设计来建立适当的 ATV 连接点、E3 连接酶招募器和 HIV-1 Gag-Pol 泛素化/蛋白酶体降解所需的接头长度和组成目标。 AIM 2 的目的是提供概念证明，证明 ATV-TPD 对野生型和 ATV 具有优于 ATV 的活性我们将系统地筛选四个系列的新型 ATV-TPD 的体外活性。单轮感染性和多轮复制测定以确定最有希望的 ATV-TPD 候选者。后者将在补充性的有限范围机制研究中进行测试，包括比较 ATV-TPD 与带有无活性 E3 连接酶配体的类似物，以探测抗病毒活性是否与提议的 MOA 一致。 IMPACT 将是针对 HIV-1 Gag-Pol 和前体 PR 的 TPD，通过限制感染性和复制一种与基于占用的 HIV-1 PI 不同的机制，这将刺激有效药物的未来发展。针对 PI 耐药 HIV 菌株的治疗方案，降低耐药发展的易感性。