Cheminformatic Discovery of Alternative Pathway C3 Pro-Convertase Inhibitors

替代途径 C3 前转化酶抑制剂的化学信息学发现

基本信息

批准号：
8877399
负责人：
Brian V Geisbrecht
金额：
$ 18.8万
依托单位：
KANSAS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8877399
关键词：
Acute Address Affect Affinity Alternative Complement Pathway Anaphylatoxins Anti-Inflammatory Agents Anti-inflammatory Area Autoimmunity Binding Biochemical Biological Assay Cell surface Chemicals Chronic Clinic Complement Complement 3b Complement Activation Complement Inactivators Complex Computer Simulation Defense Mechanisms Development Disease Drug Targeting Event Evolution FDA approved Family Foundations Future Goals Graft Rejection Health Human Human body Immune system Immunity Inflammatory Inflammatory Response Intervention Investigation Laboratories Lead Libraries Malignant Neoplasms Mediating Medical Methodology Methods Modeling Modification Natural Selections Nature Neurodegenerative Disorders Outcome Pathway interactions Peptide Hydrolases Pharmaceutical Preparations Play Population Price Process Protein Array Proteins Quantitative Structure-Activity Relationship Rare Diseases Reperfusion Injury Research Role Route Scientist Sepsis Syndrome Series Site Specificity Staging Staphylococcus aureus Structure Structure-Activity Relationship Surface System Testing Therapeutic Therapeutic Intervention Validation Virulence Virulence Factors Work alternative pathway complement C3 convertase base biochemical tools biomaterial incompatibility cheminformatics complement C3 precursor complement system driving force experience extracellular human disease inhibitor/antagonist member molecular recognition novel novel strategies pathogen pharmacophore protein complex response scaffold screening small molecule therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Although complement serves as a pillar of the innate immune system, inappropriate or uncontrolled levels of complement activation are a major contributor to a rapidly growing list of human inflammatory diseases. Yet even though therapeutic manipulation of the complement response would be tremendously important in the clinic, only a limited panel of FDA approved drugs and indications exist for specifically targeting an active component within the complement system. A major factor underlying this dearth of complement-targeted therapeutics is the large size of the proteins and complex nature of the molecular recognition events which underlie complement activity. This has made it very challenging for scientists to identify which specific points are most susceptible for therapeutic intervention. By contrast, the bacterial pathogen Staphylococcus aureus deploys an array of proteins that have been naturally selected and exquisitely optimized during the course of host/pathogen co-evolution to efficiently block fundamental events within the complement cascade - in particular formation of the multi-subunit AP C3 convertase that drives amplification of the complement response. As a consequence, molecules which mimic the activities of staphylococcal virulence proteins hold promise for complement-directed, anti-inflammatory therapeutics. We recently investigated the potential of using novel cheminformatic tools for identifying small molecules that bind the same C3b site as the SCIN family of S. aureus complement inhibitors. Although done on a pilot scale, this work strongly suggested that in silico methods are a viable means of screening for new chemotypes of drug-like compounds that specifically target functionally significant sites on C3b. In this investigation, our goal is to aply the same cheminformatic methods to screen a highly elaborated (~20 million) compound library and to identify small molecules that bind C3b sites with known roles in the initial step of AP C3 convertase formation. This step, which generates a structure known as the AP C3 pro-convertase, is the same point in the pathway that is disrupted by multiple families of secreted S. aureus complement inhibitors (including SCINs). Thus, we believe that formation of the AP C3 pro-convertase is a biologically validated target for pharmacological intervention in the complement-mediated inflammatory response. The research plan described in this application consists of three distinct specific aims. Each aim has a defined outcome that is based upon either preliminary studies directly relevant to this project or our previous experience in using similar methodologies. In the first aim, we will use cheminformatics to identify ~100 hit compounds that are likely binders at one of four target sites on C3b with known roles in AP C3 pro-convertase formation. Then, in the second aim, we will use established biochemical and functional tools to validate ~20 candidates that both bind C3b and inhibit AP activity. In the fina aim, we will use structural comparisons to identify the pharmacophores in these validated candidates and conduct a limited synthetic expansion of our most promising ~ 5 compound series. At the conclusion of this aim, we will have identified a single, non-toxic compound that will serve as our lead inhibitor of AP C3 pro-convertase assembly. As a consequence, this work will lay the foundation for future development of new classes of complement-targeted anti-inflammatory therapeutics.

描述（由申请人提供）：尽管补体是先天免疫系统的支柱，但不适当或不受控制的补体激活水平是迅速增长的人类炎症性疾病清单的主要因素。然而，即使对补体反应的治疗操作在诊所中非常重要，但只有一个有限的FDA批准的药物和适应症是有限的。补充系统中的活动组件。否定靶向疗法的这一缺乏的主要因素是蛋白质的巨大尺寸和分子识别事件的复杂性质，其基于补体活动的基础。这使科学家确定哪些特定要点最容易受到治疗干预，这使得非常具有挑战性。相比之下，细菌病原体金黄色葡萄球菌部署了一系列蛋白质，这些蛋白质已自然选择并在宿主/病原体共同进化过程中被自然地进行了优化，以有效地阻止补体级联反应中的基本事件 - 在多种益生度AP C3转换酶的特定形成中，可以使补充响应的互补响应。结果，模仿葡萄球菌毒力蛋白活性的分子对补体指导的抗炎疗法有望。最近，我们研究了使用新型化学素化工具来识别与金黄色葡萄球菌补体抑制剂相同的C3b位点结合的小分子。尽管在试验量表上完成，但这项工作强烈建议，在计算机中，方法是筛选新的类似药物样化合物的化学型的可行手段，这些化合物在C3B上特别靶向具有功能意义的位点。在这项研究中，我们的目标是使用相同的化学方法方法来筛选高度详细的（约2000万）化合物库，并识别在AP C3转化酶形成的首次步骤中结合具有已知作用的C3B位点的小分子。此步骤生成一种称为AP C3 pro-Conterventase的结构，是该途径中的相同点，被分泌的金黄色葡萄球菌补体抑制剂（包括SCIN）的多个家族中断。因此，我们认为AP C3促转换酶的形成是在补体介导的炎症反应中进行药理学干预措施的生物学验证靶标。本应用程序中描述的研究计划包括三个不同的特定目标。每个目标都有一个定义的结果，该结果基于与该项目直接相关的初步研究或我们以前使用类似方法的经验。在第一个目标中，我们将使用化学信息学识别约100种命中化合物，这些化合物可能是C3B上四个目标位点之一的粘合剂，在AP C3 pro-Convertase形成中具有已知作用。然后，在第二个目标中，我们将使用已建立的生化和功能工具来验证约有20种结合C3B并抑制AP活性的候选者。在FILA AIM中，我们将使用结构比较来识别这些经过验证的候选者中的药算归因，并对我们最有前途的〜5化合物系列进行有限的合成膨胀。在此目标的结论中，我们将确定一种无毒的化合物，该化合物将作为我们的AP C3 pro-Convertase组装的铅抑制剂。结果，这项工作将为未来的新型辅助抗炎疗法开发奠定基础。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Quantitative monitoring of two simultaneously binding species using Label-Enhanced surface plasmon resonance.

使用标记增强表面等离子体共振定量监测两个同时结合的物质。

DOI：
10.1016/j.bbrc.2018.02.040
发表时间：
2018
期刊：
Biochemical and biophysical research communications
影响因子：
3.1
作者：
Eng,Lars;Garcia,BrandonL;Geisbrecht,BrianV;Hanning,Anders
通讯作者：
Hanning,Anders

Identification of a staphylococcal complement inhibitor with broad host specificity in equid Staphylococcus aureus strains.

DOI：
10.1074/jbc.ra117.000599
发表时间：
2018-03-23
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
de Jong NWM;Vrieling M;Garcia BL;Koop G;Brettmann M;Aerts PC;Ruyken M;van Strijp JAG;Holmes M;Harrison EM;Geisbrecht BV;Rooijakkers SHM
通讯作者：
Rooijakkers SHM

Expression, purification, and characterization of a human complement component C3 analog that lacks the C-terminal C345c domain.

缺乏 C 末端 C345c 结构域的人补体成分 C3 类似物的表达、纯化和表征。