Rational design of indole compounds as HIV fusion inhibitors

作为HIV融合抑制剂的吲哚化合物的合理设计

• 批准号: 8071661
• 负责人: MIRIAM GOCHIN
• 金额: $ 20.4万
• 依托单位: TOURO UNIVERSITY OF CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-13 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8071661
• 关键词: Accounting; Affinity; Area; Binding; Binding Sites; Bioavailable; Biochemical; Biological; Biological Assay; C-Peptide; Cell fusion; Cells; Cervical; Characteristics; Chemistry; Chimeric Proteins; Complex; Data; Development; Docking; Drug Design; Drug Formulations; Drug Kinetics; FDA approved; Filovirus; Foundations; Future; Goals; HIV; HIV Fusion Inhibitors; HIV Infections; HIV-1; Half-Life; Immune; In Vitro; Individual; Indoles; Infection; Infection prevention; Inhibitory Concentration 50; Intervention; Lead; Ligand Binding; Ligands; Liquid substance; Living Costs; Location; Methodology; Methods; Modification; Molecular Bank; Molecular Weight; Paramyxovirus; Peptides; Pharmaceutical Preparations; Pharmacologic Substance; Prevalence; Procedures; Property; Prophylactic treatment; Public Health; Relaxation; Route; Screening procedure; Seminal; Seminal fluid; Series; Serum; Site; Specificity; Structure; Structure-Activity Relationship; T-20; Techniques; Testing; Therapeutic; Tropism; Viral; Virus; base; cytotoxicity; design; drug development; flexibility; high risk; improved; inhibitor/antagonist; innovation; microbicide; molecular dynamics; next generation; novel; pharmacophore; prevent; protective effect; protein protein interaction; receptor; resistant strain; restraint; scaffold; small molecule; success; tool

DESCRIPTION (provided by applicant): Development of low molecular weight HIV fusion inhibitors based on a carboxybenzyl-substituted indole scaffold is proposed. Early lead compounds show promising binding affinity to a deep hydrophobic cavity on the coiled coil of HIV-1 gp41, correlated with inhibitory activity against cell-cell and viral cell fusion. Successful inhibition of gp41 provides a universal barrier to initial infection by HIV, and has been shown to have important immune-protective effects in an infected individual by preventing the spread of infection to bystander cells. Currently, fusion inhibitors are limited to peptides derived from the gp41 C-heptad repeat, which, while highly efficacious, have pharmacological limitations such as route of administration, half-life and cost. Small molecule inhibition of HIV fusion has been a long sought after but elusive goal and success in this area would mark a significant improvement in available therapies as well as potential for prophylaxis in a microbicidal formulation. We plan synthesis of 50 - 100 new compounds to decipher the pharmacophore required to inhibit the protein - protein interaction involved in the gp41 N - C-peptide interaction, relying on relatively simple chemistry to make modifications to the basic scaffold. We have specific biochemical assays to determine affinity of our compounds for the known hydrophobic cavity in gp41, and will correlate binding activity with biological activity using cell-cell and viral cell fusion assays in the presence or absence of additives such as serum, seminal and cervical fluid. We have also developed the foundation for a novel NMR method for structure determination of bound ligands, which will be expanded for use with the high affinity ligands being developed in this proposal. From a starting compound with 55M binding affinity and 95M IC50 for fusion inhibition, we have synthesized several next generation inhibitors, and have obtained a 10-fold improvement in activity. Our goal is to obtain inhibitors that are at least 100-fold improved over the lead, having < 100nM IC50 for fusion inhibition. Thus our specific aims are 1) to utilize our current understanding of structure-activity relationships to design a new set of indole containing inhibitors, and to use binding, cell-cell fusion and viral-cell fusion assays to confirm activity, mechanism and specificity for HIV; 2) to develop NMR procedures to obtain experimental restraints for computational structure determination of bound ligands, as an aid to the rational design of improved inhibitors. Construction and testing of this molecular library promises to result in low molecular weight inhibitors with greatly improved biological activity against HIV fusion and with a well-defined mechanism of action. Such compounds could be developed into inexpensive non-peptide fusion inhibitors, which would revolutionize both treatment options and prophylaxis for the millions of new infections that occur worldwide annually. PUBLIC HEALTH RELEVANCE: The prevalence of HIV infection remains a significant public health problem due to the development of viral strains resistant to current treatments, and due to spread of the virus from individuals who are unaware they are infected. This proposal seeks to apply structure-aided drug design to develop low molecular weight HIV fusion inhibitors which prevent viral entry and the cell-to-cell spread that causes immune deficiency. Drugs in this class would be useful in microbicides to prevent infection, or could be given orally to treat infection as an alternative to the currently used fusion inhibitor Enfuvirtide(R), which must be given intravenously.

描述（由申请人提供）：建议开发基于羧基苄基取代的吲哚支架的低分子量HIV融合抑制剂。早期的先导化合物对 HIV-1 gp41 卷曲线圈上的深层疏水空腔表现出良好的结合亲和力，这与细胞间和病毒细胞融合的抑制活性相关。成功抑制 gp41 为 HIV 初始感染提供了普遍屏障，并且已被证明可以通过防止感染传播到旁观者细胞来对感染个体产生重要的免疫保护作用。目前，融合抑制剂仅限于源自 gp41 C-七肽重复序列的肽，其虽然非常有效，但具有药理学限制，例如给药途径、半衰期和成本。 HIV融合的小分子抑制一直是人们长期追求但难以捉摸的目标，该领域的成功将标志着现有疗法的显着改进以及杀菌制剂的预防潜力。我们计划合成 50 - 100 种新化合物，以破译抑制 gp41 N - C 肽相互作用中涉及的蛋白质 - 蛋白质相互作用所需的药效团，依靠相对简单的化学对基本支架进行修饰。我们通过特定的生化测定来确定我们的化合物对 gp41 中已知疏水腔的亲和力，并在存在或不存在血清、精液和宫颈液等添加剂的情况下，使用细胞-细胞和病毒细胞融合测定将结合活性与生物活性相关联。我们还开发了一种用于确定结合配体结构的新型核磁共振方法的基础，该方法将扩展到与本提案中正在开发的高亲和力配体一起使用。从具有 55M 结合亲和力和 95M IC50 融合抑制的起始化合物，我们合成了几种下一代抑制剂，并且活性提高了 10 倍。我们的目标是获得比先导抑制剂至少提高 100 倍的抑制剂，其融合抑制 IC50 < 100nM。因此，我们的具体目标是1）利用我们目前对结构-活性关系的理解来设计一套新的含吲哚抑制剂，并使用结合、细胞-细胞融合和病毒-细胞融合测定来确认HIV的活性、机制和特异性； 2) 开发核磁共振程序以获得结合配体的计算结构测定的实验限制，作为改进抑制剂的合理设计的辅助。该分子库的构建和测试有望产生低分子量抑制剂，其抗 HIV 融合的生物活性大大提高，并且具有明确的作用机制。此类化合物可以开发成廉价的非肽融合抑制剂，这将彻底改变全球每年发生的数百万新感染的治疗选择和预防。 公共卫生相关性：艾滋病毒感染的流行仍然是一个重大的公共卫生问题，因为病毒株对目前的治疗方法产生了抗药性，而且病毒是从不知道自己已被感染的个体中传播的。该提案旨在应用结构辅助药物设计来开发低分子量 HIV 融合抑制剂，以防止病毒进入和导致免疫缺陷的细胞间传播。此类药物可用于杀微生物剂以预防感染，或者可以口服给药来治疗感染，作为目前使用的必须静脉内给药的融合抑制剂Enfuvirtide 的替代品。