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）：虽然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导致人类炎症性疾病快速增长的主要原因。然而，尽管补体反应的治疗操作在临床上非常重要，但只有有限的 FDA 批准的药物和适应症用于专门针对补体系统内的活性成分。缺乏补体靶向治疗的一个主要因素是蛋白质的大尺寸和作为补体活性基础的分子识别事件的复杂性。这使得科学家很难确定哪些特定点最容易受到治疗干预。相比之下，细菌病原体金黄色葡萄球菌部署了一系列在宿主/病原体共同进化过程中自然选择和精心优化的蛋白质，以有效阻止补体级联内的基本事件 - 特别是多亚基 AP 的形成C3 转化酶驱动补体反应放大。因此，模仿葡萄球菌毒力蛋白活性的分子有望用于补体导向的抗炎治疗。我们最近研究了使用新型化学信息学工具来识别与金黄色葡萄球菌补体抑制剂 SCIN 家族结合相同 C3b 位点的小分子的潜力。尽管是中试规模，但这项工作强烈表明，计算机模拟方法是筛选专门针对 C3b 上功能重要位点的类药化合物新化学型的可行方法。在这项研究中，我们的目标是应用相同的化学信息学方法来筛选高度精细的（约 2000 万）化合物库，并鉴定在 A C3 转化酶形成的初始步骤中与已知作用的 C3b 位点结合的小分子。这一步骤会生成一种称为 AP C3 前转化酶的结构，该步骤与被分泌性金黄色葡萄球菌补体抑制剂（包括 SCIN）的多个家族破坏的途径中的同一点相同。因此，我们相信，AP C3 转化酶原的形成是补体介导的炎症反应中药物干预的经过生物学验证的靶点。本申请中描述的研究计划包含三个不同的具体目标。每个目标都有一个明确的结果，该结果基于与该项目直接相关的初步研究或我们之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来识别约 100 种命中化合物，这些化合物可能是 C3b 上四个靶位点之一的结合物，在 AP C3 前转化酶形成中具有已知的作用。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 20 种既能结合 C3b 又能抑制 AP 活性的候选药物。在最终目标中，我们将使用结构比较来识别这些经过验证的候选药物中的药效团，并对我们最有前途的约 5 种化合物系列进行有限的合成扩展。完成这一目标后，我们将鉴定出一种单一的无毒化合物，它将作为我们的 AP C3 前转化酶组装体的主要抑制剂。因此，这项工作将为未来开发新型补体靶向抗炎疗法奠定基础。

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。