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.

项目摘要 为全球3790万艾滋病毒携带者提供功能性治疗的首要障碍是 消除HIV-1病毒的宿主。病毒整合酶(IN)在转录不活跃的BUT中建立终身感染 IN通过将前病毒DNA整合到宿主染色体中来复制有能力的细胞。的本土化 宿主因子LEDGF/p75与活性转录基因密集区结合的整合 关于DNA的。HIV-1 IN是主要的抗病毒靶点之一，目前已开发出多种IN抑制剂(INST) 作为目前抗逆转录病毒疗法(ART)的关键组成部分，但受到病毒耐药性、交叉耐药性的影响 和毒性。高基因屏障、新作用模式、安全的抗病毒药物的快速发展 我们迫切需要DMPK配置文件。变构IN抑制剂(ALLINI)是一类新的靶向IN抑制剂 HIV-1的非催化部位。STP0404是一种以吡咯吡啶为基础的Allini，具有显著的疗效、PK和 在临床前体外和动物研究中观察到的安全性概况。STP0404成为人类第一人 (FIH)Allini化合物显示其卓越的安全性和人类一天一次口服的PK特征 这进一步支持对其在HIV-1患者中的抗病毒疗效的高级临床评估。至 这种效果，我建议探索一种新的第二代Allini化合物，我们的实验室设计的潜在 改善了遗传屏障，并可能用作HIV-1治愈剂。我在突变渲染中发现了一种128T 病毒对STP0404的抵抗力，我们的X射线结构研究证实了A128T碰撞的较长侧链 针对STP0404的吡咯部分的3-甲基，有助于降低与STP0404的结合亲和力 以及对STP0404的病毒抗性。为了避免A128T突变体与STP0404的甲基接触， 本课题组最近合成了一种新的缺少甲基的STP0404衍生物EKC110 A128T侧链。在这里，我建议检验EKC110显示出改进的遗传屏障的假设 与亲本STP0404相比，耐药突变谱发生了变化。我们的初步数据显示 支持这一假设，此外，新出现的证据表明，Allini阻断了LEDGF/p75的结合 TO IN，称为LEDGINs，可以将HIV-1的整合位点特异性驱动到非活跃转录区域 减少潜伏感染的T细胞库中HIV-1的重新激活。在《目标1》中，我将 应用分子、病毒学和细胞生物学技术研究新的STP0404衍生物EKC110 基因屏障、病毒耐药性、抗病毒MOA确定结构修饰对HIV-1的影响 抗病毒活性。目的研究STP0404和EKC110在HIV-1整合位点选择中的作用 利用生化、分子和细胞生物学技术抑制HIV-1潜伏感染T细胞的再激活 技巧。总而言之，这些目标将证明我们的基于吡咯吡啶的ALLINI，如 EKC110可作为一种新的抗逆转录病毒药物，不仅能改善遗传屏障，而且还能抑制HIV的重新激活 作为一种“阻断和锁定”试剂的能力，既可用于病毒感染，也可用于持久性。