PROTACS Against Nef as a Functional Cure for HIV Infection

PROTACS 针对 Nef 作为 HIV 感染的功能性治疗

• 批准号: 10079715
• 负责人: Thomas E. Smithgall
• 金额: $ 30万
• 依托单位: FOX CHASE CHEMICAL DIVERSITY CENTER, INC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10079715
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Active Sites; Affinity; Anti-Retroviral Agents; Antigen Presentation; Autologous; Binding; Binding Proteins; Biological Assay; Biological Availability; CD4 Positive T Lymphocytes; Caco-2 Cells; Cell surface; Cells; Cellular Assay; Chemicals; Chimera organism; Clinical Research; Coupled; Cytotoxic T-Lymphocytes; Data; Development; Down-Regulation; Drug Industry; Drug Kinetics; Drug Targeting; Foundations; Foxes; Future; Goals; HIV; HIV Infections; HIV-1; Half-Life; Human; Ice; Immune; Immune system; In Vitro; Individual; Laboratories; Libraries; Life; Life Cycle Stages; Ligand Binding; Ligands; Literature; Liver Microsomes; Mediating; Metabolic; Midazolam; Molecular Weight; Mus; Oral; Patients; Peripheral Blood Mononuclear Cell; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Phase; Plasma Proteins; Polyubiquitination; Positioning Attribute; Program Development; Property; Proteins; Recombinants; Relapse; Reporting; Research; Response Latencies; Role; Series; Small Business Technology Transfer Research; Solubility; Structure; Surface Plasmon Resonance; System; Testing; Therapeutic; Toxicology; Universities; Viral; Viral reservoir; Virulence Factors; Work; antiretroviral therapy; aqueous; base; cell type; clinical translation; cytotoxicity; drug candidate; drug development; efficacy evaluation; flexibility; humanized mouse; in vivo; in vivo evaluation; inhibitor/antagonist; innovation; insight; latent HIV reservoir; mouse model; nef Protein; novel; novel strategies; novel therapeutic intervention; phase 1 study; pre-clinical; prevent; response; small molecule; therapeutic target; ubiquitin-protein ligase; AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Active Sites; Affinity; Anti-Retroviral Agents; Antigen Presentation; Autologous; Binding; Binding Proteins; Biological Assay; Biological Availability; CD4 Positive T Lymphocytes; Caco-2 Cells; Cell surface; Cells; Cellular Assay; Chemicals; Chimera organism; Clinical Research; Coupled; Cytotoxic T-Lymphocytes; Data; Development; Down-Regulation; Drug Industry; Drug Kinetics; Drug Targeting; Foundations; Foxes; Future; Goals; HIV; HIV Infections; HIV-1; Half-Life; Human; Ice; Immune; Immune system; In Vitro; Individual; Laboratories; Libraries; Life; Life Cycle Stages; Ligand Binding; Ligands; Literature; Liver Microsomes; Mediating; Metabolic; Midazolam; Molecular Weight; Mus; Oral; Patients; Peripheral Blood Mononuclear Cell; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Phase; Plasma Proteins; Polyubiquitination; Positioning Attribute; Program Development; Property; Proteins; Recombinants; Relapse; Reporting; Research; Response Latencies; Role; Series; Small Business Technology Transfer Research; Solubility; Structure; Surface Plasmon Resonance; System; Testing; Therapeutic; Toxicology; Universities; Viral; Viral reservoir; Virulence Factors; Work; antiretroviral therapy; aqueous; base; cell type; clinical translation; cytotoxicity; drug candidate; drug development; efficacy evaluation; flexibility; humanized mouse; in vivo; in vivo evaluation; inhibitor/antagonist; innovation; insight; latent HIV reservoir; mouse model; nef Protein; novel; novel strategies; novel therapeutic intervention; phase 1 study; pre-clinical; prevent; response; small molecule; therapeutic target; ubiquitin-protein ligase

Abstract. Existing antiretroviral drugs do not clear HIV-1 latent reservoirs, underscoring the urgent need for new therapeutic strategies. The HIV-1 Nef accessory factor is an attractive target for drug development because of its critical roles in the HIV-1 life cycle and immune system escape. Our group has discovered novel small molecules that bind directly to Nef and block many of its functions, including enhancement of viral infectivity and replication in donor PBMCs. Importantly, our Nef inhibitors rescue cell-surface MHC-I expression in latently in- fected, patient-derived CD4 T-cells, enabling recognition and killing by autologous CTLs. Thus, Nef inhibitors represent an innovative approach to antiretroviral therapy that may provide a path to eradication of viral reser- voirs. Our most promising class of inhibitors (hydroxypyrazoles) bind tightly to their Nef protein target in vitro with KD values in the nM to pM range. However, Nef lacks an active site, which has complicated traditional medicinal chemistry optimization of existing compounds for in vivo testing due to the lack of correlation between Nef binding affinity in vitro and antiretroviral activity in cell-based systems. To circumvent this issue, we propose to use our existing Nef-binding compounds to develop Proteolytic Targeting Chimera (PROTAC) molecules for the targeted destruction of the Nef protein in HIV-infected cells. In this approach, existing hydroxypyrazole Nef-binding compounds will be coupled to ligands for ubiquitin E3 lig- ases via a flexible linker. The resulting Nef PROTACs are anticipated to catalyze the proteolytic degradation of Nef via E3-mediated polyubiquitination and proteasomal targeting. A major advantage of the PROTAC approach is that it requires only a selective binder of the target protein (Nef in this case) and not a functional inhibitor. We anticipate that selective PROTAC-mediated degradation will eliminate all Nef functions, including rescue of cell surface MHC-I mediated HIV-1 antigen presentation, leading to clearance of HIV+ cells via the CTL response as part of a strategy for latent reservoir reduction and functional cure. More generally, the PROTAC approach has generated a great deal of excitement in the pharmaceutical industry, because it provides new avenues to inhibit proteins previously considered undruggable. While PROTACs have higher molecular weights that typical small molecule drugs, recent preclinical and Phase I clinical studies have demonstrated activity in vivo as well as oral bioavailability. For this Phase I STTR project, HIV-1 Nef PROTAC development will combine the pharmaceutical and medicinal chemistry expertise of the Fox Chase Chemical Diversity Center, Inc. (www.fc-cdci.com; FCCDC) with the expertise of the Smithgall Lab at the University of Pittsburgh in HIV-1 Nef structure, function and inhibitor analysis. Our broad goal is to synthesize and test a series of Nef PROTACs with different Nef-targeting moieties, linkers, and E3 ubiquitin ligase ligands to identify compounds suitable for in vivo proof-of-concept studies. Suc- cessful completion of Phase I will provide a strong foundation for an expanded drug development program in Phase II, with the ultimate goal of clinical translation.

抽象的。现有的抗逆转录病毒药物尚未清除HIV-1潜在储层，强调了迫切需要 新的治疗策略。 HIV-1 NEF附件因子是药物开发的有吸引力的目标，因为 它在HIV-1生命周期和免疫系统中逃脱的关键作用。我们的小组发现了小说 直接与NEF结合并阻止其许多功能的分子，包括增强病毒感染性和 供体PBMC中的复制。重要的是，我们的NEF抑制剂在潜在的内部挽救细胞表面MHC-I表达 经过的患者衍生的CD4 T细胞，可自体CTL识别并杀死。因此，NEF抑制剂 代表一种创新的抗逆转录病毒疗法的方法，该方法可能为根除病毒的途径提供途径 瞧。我们最有前途的抑制剂（羟基吡唑）在体外与Nef蛋白靶标紧密结合 在NM到PM范围内具有KD值。但是，NEF缺乏一个活跃的网站，这使传统的传统网站复杂复杂 由于缺乏相关性，用于体内测试现有化合物的药物化学优化 基于细胞系统的体外和抗逆转录病毒活性的NEF结合亲和力。 为了解决这个问题，我们建议使用现有的NEF结合化合物开发蛋白水解 靶向嵌合体（Protac）分子，以靶向HIV感染细胞中NEF蛋白的靶向破坏。在 这种方法，现有的羟基吡唑NEF结合化合物将与配体偶联用于泛素E3 Lig- 通过灵活的链接器ASE。预计所得的NEF Protac将催化 通过E3介导的多泛素化和蛋白酶体靶向NEF。 Protac方法的主要优势 是它仅需要靶蛋白的选择性粘合剂（在这种情况下为NEF），而不需要功能抑制剂。我们 预计选择性的Protac介导的降解将消除所有NEF功能，包括拯救细胞 表面MHC-1介导的HIV-1抗原表现，导致通过CTL响应清除HIV+细胞 潜在储层还原和功能治疗的策略的一部分。更一般地，Protac方法具有 在制药行业引起了极大的兴奋，因为它提供了抑制的新途径 蛋白质以前认为不良。虽然protac的分子量较高，而典型的小 分子药物，最近的临床前和I期临床研究表明在体内和口腔中的活性 生物利用度。对于此I阶段ISTTR项目，HIV-1 NEF Protac开发将结合药物 和Fox Chase化学多样性中心的药物化学专业知识（www.fc-cdci.com; fccdc） 匹兹堡大学HIV-1 NEF结构，功能和抑制剂的Smithgall实验室的专业知识 分析。我们的广泛目标是综合和测试一系列具有不同靶向NEF的部分的NEF Protac， 接头和E3泛素连接酶配体识别适合体内概念验证研究的化合物。成功 第一阶段的结束将为扩大的药物开发计划提供坚实的基础 第二阶段，是临床翻译的最终目标。