Anti-HIV and anti-reactivation activities of pyrrolopyridine-based ALLINIs

基于吡咯并吡啶的 ALLINI 的抗 HIV 和抗再激活活性

• 批准号: 10762564
• 负责人: LINDSEY RAMIREZ
• 金额: $ 5.02万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10762564
• 关键词: Affinity; Animals; Anti-HIV Agents; Antiviral Agents; Binding; Binding Sites; Biochemical; Biochemistry; Biological; Biological Assay; Capsid; Cells; Chemicals; Chromosomes; Clinical; DNA; Data; Development; Electron Microscopy; Evaluation; Formulation; Future; Generations; Genetic; Genetic Transcription; Genome; Goals; HIV; HIV-1; HIV-1 integrase; Human; In Vitro; Infection; Integrase; Integrase Inhibitors; Integration Host Factors; Investigation; Knowledge; Laboratories; Mediating; Modification; Molecular; Oral; Patients; Persons; Pharmacologic Substance; Pyrroles; Reporting; Resistance; Roentgen Rays; Safety; Series; Side; Site; Specificity; T-Lymphocyte; Techniques; Testing; Toxic effect; Toxicology; Viral; Viral Physiology; Viral reservoir; Virus; Virus Diseases; Virus Integration; anti-viral efficacy; antiretroviral therapy; clinical development; design; first-in-human; improved; inhibitor; integration site; methyl group; monomer; mutant; phase I trial; pre-clinical; promoter; pyridine; quinoline; reactivation from latency; research clinical testing; resistance mutation; safety study; scaffold; structural biology; transcription factor; transcriptional coactivator p75; viral RNA; viral resistance; virology

Project Summary The overarching obstacle in obtaining a functional cure for the 37.9 millions of people living with HIV globally is eliminating the HIV-1 reservoirs. Viral integrase (IN) establishes lifelong infection in transcriptionally inactive but replication competent cells by IN integrating proviral DNA into the host chromosome. The localization of integration is directed by host factor LEDGF/p75 binding IN tethering to active transcriptional gene dense regions of DNA. HIV-1 IN is one of the major antiviral targets, and multiple IN inhibitors (INSTs) have been developed as a key component of current antiretroviral therapy (ART) but are subjected to viral resistance, cross-resistance and toxicity. Rapid development of IN antivirals with high genetic barrier, new modes of action (MOA), with safe DMPK profiles are in immediate need. Allosteric IN inhibitors (ALLINIs) are a new class of IN inhibitors targeting non-catalytic sites of HIV-1 IN. STP0404 is a pyrrolopyridine-based ALLINI with outstanding efficacy, PK and safety profiles observed in both preclinical in vitro and animal investigations. STP0404 became the first-in-human (FIH) ALLINI compound demonstrating its outstanding safety and human PK profile for once-a-day oral formulations, which further supports advanced clinical evaluations for its antiviral efficacy in HIV-1 patients. To this effect, I propose to explore a new 2nd-generation ALLINI compound our laboratory designed for potentially improved genetic barrier and possible utility as an HIV-1 cure agent. I have identified A128T IN mutant rendering viral resistance to STP0404, and our X-ray structural study confirmed the longer side chain of A128T clashes against 3-methyl group at the pyrrole moiety of STP0404, contributing to the reduced binding affinity to STP0404 and viral resistance to STP0404. To avoid contact between the A128T mutant and the methyl group of STP0404, our group recently synthesized a new STP0404 derivative, EKC110, with missing the methyl group contacting the A128T side chain. Here, I propose to test the hypothesis that EKC110 shows improved genetic barrier with altered resistance mutation profiles, compared to the parental STP0404. Our preliminary data has supported this hypothesis, additionally emerging evidence has reported ALLINIs blocking LEDGF/p75 binding to IN, called LEDGINs, can drive the integration site specificity of HIV-1 toward non-actively transcribed regions of the chromosomes, reducing HIV-1 reactivation from latently infected T cell reservoirs. In Aim 1, I will investigate a new STP0404 derivative, EKC110 using molecular, virological and cell biological techniques for its genetic barrier, viral resistance, antiviral MOA to determine the impact of structural modifications on HIV-1 antiviral activity. Aim 2, I will investigate the effect of STP0404 and EKC110 on HIV-1 integration site selection and reactivation suppression from HIV-1 latently infected T cells using biochemical, molecular, and cell biological techniques. Collectively these aims will demonstrate the utility that our pyrrolopyridine based ALLINIs such as EKC110 can be a new ART agent with not only improved genetic barrier but also HIV reactivation inhibition capability as a “Block and Lock” agent, which can be used for both viral infection and persistence.

项目概要 全球 3790 万艾滋病毒感染者获得功能性治愈的首要障碍是 消除 HIV-1 病毒库。病毒整合酶（IN）在转录不活跃但不活跃的细胞中建立终生感染 通过将原病毒 DNA 整合到宿主染色体中，复制能力细胞。本地化 整合是由宿主因子 LEDGF/p75 结合束缚到活性转录基因密集区域来指导的 DNA。 HIV-1 IN是主要抗病毒靶点之一，多种IN抑制剂（INST）已被开发 作为当前抗逆转录病毒疗法（ART）的关键组成部分，但面临病毒耐药性、交叉耐药性 和毒性。快速开发具有高遗传屏障、新作用模式（MOA）、安全的IN抗病毒药物 DMPK 配置文件是迫切需要的。变构 IN 抑制剂 (ALLINI) 是一类新型 IN 抑制剂，靶向 HIV-1 IN 的非催化位点。 STP0404是一种基于吡咯并吡啶的ALLINI，具有出色的功效、PK和 在临床前体外和动物研究中观察到的安全性概况。 STP0404成为人类首个 (FIH) ALLINI 化合物每日口服一次，展示其出色的安全性和人体 PK 特性 制剂，进一步支持其对 HIV-1 患者的抗病毒功效的高级临床评估。到 对于这种效应，我建议探索一种新的第二代 ALLINI 化合物，我们的实验室设计用于潜在的 改善了遗传屏障，并可能作为 HIV-1 治疗剂使用。我已经在突变体渲染中识别出 A128T 病毒对 STP0404 具有抗性，我们的 X 射线结构研究证实了 A128T 的较长侧链发生冲突 针对 STP0404 吡咯部分的 3-甲基，导致与 STP0404 的结合亲和力降低 以及病毒对 STP0404 的耐药性。为了避免 A128T 突变体和 STP0404 的甲基之间的接触， 我们小组最近合成了一种新的STP0404衍生物EKC110，缺少甲基接触 A128T侧链。在这里，我建议检验 EKC110 显示出改善的遗传屏障的假设 与亲本 STP0404 相比，具有改变的抗性突变谱。我们的初步数据有 支持这一假设，另外新出现的证据表明 ALLINI 阻断 LEDGF/p75 结合 IN，称为 LEDGIN，可以驱动 HIV-1 向非活跃转录区域整合位点特异性 染色体，减少潜伏感染 T 细胞储存库中 HIV-1 的重新激活。在目标 1 中，我将 使用分子、病毒学和细胞生物学技术研究一种新的 STP0404 衍生物 EKC110 遗传屏障、病毒耐药性、抗病毒 MOA 以确定结构修饰对 HIV-1 的影响 抗病毒活性。目标2，我将研究STP0404和EKC110对HIV-1整合位点选择的影响 使用生化、分子和细胞生物学方法抑制 HIV-1 潜伏感染 T 细胞的再激活 技术。总的来说，这些目标将证明我们基于吡咯并吡啶的 ALLINI 的实用性，例如 EKC110可以成为一种新的ART药物，不仅可以改善遗传屏障，还可以抑制HIV再激活 作为“阻断和锁定”剂的能力，可用于病毒感染和持久性。