Mechanism of indole compounds as HIV fusion inhibitors

吲哚类化合物作为HIV融合抑制剂的机制

基本信息

批准号：
9212779
负责人：
MIRIAM GOCHIN
金额：
$ 17.88万
依托单位：
TOURO UNIVERSITY OF CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-02-01 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9212779
关键词：
Anti-Retroviral Agents Binding Binding Sites Biological Availability Biophysics C-Peptide Carrier Proteins Cell Nucleus Cell fusion Cell membrane Cells Characteristics Charge Chimeric Proteins Cocrystallography Complex Crystallization Crystallography Data Set Development Disease Progression Ensure Environment Glycoproteins Goals Grant HIV HIV Fusion Inhibitors HIV-1 Hydrophobicity Immune Indoles Infection Interruption Knowledge Label Ligands Link Lipids Maleimides Membrane Methodology Methods Mission Modeling Modification Molecular Molecular Conformation Molecular Weight Nature Peptides Pharmaceutical Preparations Phase Predisposition Proteins Proteolysis Resistance Resistance profile Resolution Resource Development Scheme Series Site Solubility Structure Techniques Testing Therapeutic Time United States National Institutes of Health Viral Viral Fusion Proteins Virus Work base cost design drug discovery experimental study globular protein human disease improved inhibitor/antagonist microbicide mimetics neutralizing antibody novel parenteral administration polypeptide C prevent protein complex protein protein interaction public health relevance receptor scaffold screening small molecule small molecule inhibitor success technique development tool transmission process viral rebound

项目摘要

DESCRIPTION (provided by applicant): This proposal describes a ligand and structure-based approach to discovery and development of low molecular weight gp41 inhibitors effective against Human Immunodeficiency Virus (HIV-1) fusion. Fusion inhibitors possess excellent characteristics for interrupting HIV transmission and preventing disease progression, since they block initial infection of healthy cells and cell-to-cell spread of infection. Yet there are currenly no highly potent small molecule inhibitors of fusion, and peptide fusion inhibitors possess some undesirable characteristics such as high cost and susceptibility to proteolysis. Our central hypothesis is that potent small molecule fusion inhibitors targeting a known hydrophobic pocket can be rationally designed by understanding the molecular features contributing to potency and evaluating the binding mode. The objective of this application is to study and modify promising small molecules to find conditions for determining their structure bound to gp41. We have developed hydrophobic pocket binding indole compounds with IC50's down to 200nM for inhibition of virus - cell and cell - cell fusion. We have also observed that excess negative charge has a deleterious effect on the inhibitors, which we attribute to the effect of the adjacent plasma membrane. We have developed novel reverse hairpin proteins (RH's) as suitable receptors to use in protein - ligand structure studies. They present exposed hydrophobic pocket binding grooves in solution. The goals of the application are to modify the non-peptide fusion inhibitors to assess the determinants of potency, and to identify conditions for studying high resolution structural details of their complexes with gp41. We will accomplish our goals in three Specific Aims: (1) Perform ligand-based design on indole inhibitors, using SAR to predict reliable binding poses; (2) Determine conditions for crystallizing indole inhibitors with RH's to obtain high resolution structures of the complexes; (3) Determine conditions for studying RH - ligand complexes by NMR, using carrier proteins to ensure solubility of the ligands if necessary, and incorporating 19F nuclei as sensitive probes of the environment and interactions. The combination of these experiments will yield an improved understanding of the interaction between small molecule inhibitors and gp41, and the methodology to study them in detail. The significance of this proposal lies in its potential to contribute specifically to HIV-1 fusion inhiitor development and also to the development of techniques and resources applicable to other Class 1 viral fusion proteins, a broader goal relevant to NIH's mission of enhancing fundamental knowledge to treat human disease. The long-term goal of the project is to generate drug-like compounds active against HIV-1 fusion, suitable for therapeutic or microbicide use, which could be used to counter or prevent the millions of new infections that occur annually worldwide. Such compounds could have the added benefit of suppressing viral rebound and overcoming the virus' defenses against neutralizing antibodies.

描述（由适用提供）：该提案描述了一种基于配体和结构的方法，用于发现和开发低分子量的GP41抑制剂对人免疫缺陷病毒（HIV-1）融合有效。融合抑制剂具有中断HIV传播和预防疾病进展的良好特征，因为它们阻止了健康细胞的初始感染和感染细胞间传播。然而，没有高潜在的小分子融合分子抑制剂，肽融合抑制剂的潜在某些不良特征，例如高成本和蛋白水解的易感性。我们的中心假设是，靶向已知疏水口袋的潜在小分子融合抑制剂可以通过理解有助于效力和评估结合模式的分子特征来合理设计。该应用的目的是研究和修改诺言，以找到确定其结构与gp41结合的条件。我们已经开发了疏水袋结合吲哚化合物，IC50的降低至200nm，以抑制病毒 - 细胞和细胞 - 细胞融合。我们还观察到，超过负电荷对抑制剂具有有害作用，我们将其归因于相邻的影响质膜。我们已经开发了新型的反向发夹蛋白（RH）作为用于蛋白质配体结构研究的合适受体。他们在溶液中提出了暴露的疏水袋结合凹槽。该应用的目标是修改非肽融合抑制剂以评估效力的决定剂，并确定研究其与GP41复合物的高分辨率结构细节的条件。我们将在三个特定目标中实现我们的目标：（1）使用SAR来预测可靠的结合位置，对吲哚抑制剂进行基于配体的设计；（2）确定用RH的结晶吲哚抑制剂的条件，以获得配合物的高分辨率结构；（3）确定使用NMR研究RH -配体复合物的条件，使用载体蛋白在必要时确保配体的溶液，并将19F核纳入环境和相互作用的敏感问题。这些实验的组合将提高人们对小分子抑制剂与GP41之间相互作用的了解，以及详细研究它们的方法。该提案的重要性在于它有可能专门为HIV-1融合抑制剂开发以及适用于其他1类病毒融合蛋白的技术和资源的开发而做出贡献，这是与NIH增强治疗人类疾病的基本知识有关的更广泛的目标。该项目的长期目标是针对HIV-1融合产生类似药物的化合物，适合于治疗或杀生型融合，可用于抵抗或防止全球每年发生的数百万个新感染。这样的化合物可能具有抑制病毒反弹并克服中和抗体的防御能力的额外好处。