Inhibitors of Human Factor XIIIa as New Anticoagulants

人类因子 XIIIa 抑制剂作为新型抗凝剂

• 批准号: 10629057
• 负责人: Rami A Al-Horani
• 金额: $ 14.9万
• 依托单位: XAVIER UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-17 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629057
• 关键词: Active Sites; Affect; African American population; Alkanesulfonates; Allosteric Site; Animal Model; Anions; Anticoagulants; Antiplasmin; Area; Binding; Binding Sites; Biochemical; Biological; Biological Assay; Blood; Blood Coagulation Factor; Blood coagulation; Body Weight decreased; Cardiovascular Diseases; Cell Line; Cessation of life; Clinical; Coagulation Process; Contracts; Data Reporting; Deposition; Disease; Economics; Erythrocytes; Evaluation; Exhibits; Factor XIIIa; Factor Xa; Fibrin; Fibrinogen; Fibrinolysis; Future; Genetic Polymorphism; Glutaminase; Glycosaminoglycans; Goals; Hemorrhage; Hemostatic function; Heterozygote; Human; In Vitro; Investigation; Lead; Libraries; Mediating; Meta-Analysis; Minority; Morbidity - disease rate; Mus; Mutation; NOR Mouse; Naphthalene; Nature; Normal Range; Pathologic; Pathology; Patients; Peptide Hydrolases; Persons; Pharmacologic Substance; Phenocopy; Physiology; Plasmin; Polymers; Prevalence; Prevention; Property; Public Health; Recurrence; Recurrent disease; Regulation; Research; Risk; Role; Route; Serine Protease; Specificity; Structure; Therapeutic; Thrombin; Thrombus; Transglutaminases; Venous; Wild Type Mouse; chemical synthesis; clinically relevant; crosslink; design; economic impact; effective therapy; experimental study; improved; inhibitor; innovation; interdisciplinary approach; mimetics; molecular modeling; mouse model; new technology; new therapeutic target; novel strategies; pharmacologic; polymerization; prevent; therapeutic target; tool; venous thromboembolism

SUMMARY The long-term goal of our research is to develop effective anticoagulants that do not cause bleeding complications to be safely used for a wider range of patients suffering from venous thromboembolism (VTE). This project aims at developing effective and safer anticoagulants by targeting human factor XIIIa (FXIIIa). All available anticoagulants are associated with a significant risk of bleeding. Current anticoagulants inhibit directly or indirectly thrombin and/or factor Xa. This is the reason why they are clinically effective, but it is also the reason why they cause bleeding. The central hypothesis is that inhibiting FXIIIa will result in effective protection against VTE without causing significant bleeding. In contrast to all other clotting factors which are serine proteases, FXIIIa is a transglutaminase that catalyzes the last step in the coagulation process. This unique biochemical aspect of FXIIIa has been under investigation in the context of VTE. In vitro experiments showed that treating normal human blood with an experimental transglutaminase inhibitor increases RBC extrusion from contracting clots and reduces clot size. Various studies also suggested that a certain FXIIIa polymorphism provides significant protection against VTE and that heterozygous FXIII-deficient mice do not show signs of excessive bleeding. Thus, FXIIIa may serve as a potential therapeutic target to develop a new effective treatment for VTE that does not significantly increase the bleeding risk. Despite this promise, very few FXIIIa inhibitors have been developed, all of which lack substantial selectivity as they can also inhibit other transglutaminases by blocking their active sites. Thus, I have proposed sulfonated non-saccharide glycos- aminoglycan mimetics as a platform to develop FXIIIa inhibitors. The sulfonated molecules are to inhibit FXIIIa potently and selectively through allosteric modulation. In preliminary studies, I discovered two sulfonated molecules that inhibit FXIIIa with low micromolar potencies. The two molecules inhibited FXIIIa-mediated polymerization of fibrin. The two molecules did not affect other clotting factors and did not affect the viability of three cell lines. Molecular modeling projected a plausible binding site for these molecules on FXIIIa. In this proposal, I specifically aim at using a multidisciplinary approach to establish the principles of effective and selective inhibition of FXIIIa by sulfonated molecules. I will synthesize advanced libraries of two “lead” molecules and evaluate their biochemical and biological potential as anticoagulants. The proposal is innovative because i) it puts forward a novel approach to overcome the limitations of current VTE treatment; ii) it exploits a multidisciplinary approach to investigate the specific aims; and iii) it introduces new technologies with proprietary structural and mechanistic aspects. The project is also significant because it will: i) identify 2-3 potent, specific, and allosteric FXIIIa inhibitors for future evaluation in animal models of VTE and bleeding; ii) offer new tools to better understand FXIIIa role in the coagulation physiology and pathology; iii) investigate an alternative approach to modulate FXIIIa via allostery to pave the way to transforming anticoagulants.

概括 我们研究的长期目标是开发不会引起出血的有效抗凝剂 应当安全地用于患有静脉血栓栓塞（VTE）的更广泛的患者。 该项目旨在通过针对人类因素XIIIA（FXIIIA）来开发有效，更安全的抗凝剂。全部 可用的抗凝剂与出血的重大风险有关。电流抗凝剂直接抑制 或间接凝血酶和/或因子XA。这就是它们在临床上有效的原因，但也是 原因引起出血的原因。中心假设是抑制FXIIIA将导致有效 防止VTE而不会引起大量出血。与所有其他服装因素相反 丝氨酸蛋白酶，FXIIIA是一种转谷氨酰胺酶，催化凝结过程的最后一步。这 在VTE的背景下，FXIIIA的独特生化方面一直在研究。体外实验 表明，使用实验性转谷氨酰胺酶抑制剂治疗正常的人体血液会增加RBC 从收缩部分挤出并减少凝块大小。各种研究还表明某个FXIIIA 多态性为VTE提供了重要的保护，并且杂合FXIII缺陷小鼠没有 显示出血过多的迹象。这，FXIIIA可能是开发新的治疗靶标的 VTE的有效治疗方法不会显着增加出血风险。尽管有这样的承诺，很少 已经开发了FXIIIA抑制剂，所有这些抑制剂都缺乏可观的选择性，因为它们也可以抑制其他 通过阻断其活性位点的转谷氨酰胺酶。那我提出了磺化的非糖糖 - aminoglycan Mimetics作为开发FXIIA抑制剂的平台。磺化分子将抑制FXIIIA 通过变构调制潜在，有选择地。在初步研究中，我发现了两个磺化 抑制低微摩尔电位的FXIIIA的分子。这两个分子抑制了FXIIIA介导的 纤维蛋白的聚合。这两个分子不影响其他凝血因子，也不影响 三个细胞系。分子建模投影了这些分子在FXIIA上的合理结合位点。在这个 提案，我特别旨在使用多学科的方法来建立有效的原则和 通过磺化分子对FXIIA的选择性抑制。我将合成两个“铅”的高级库 分子并评估它们作为抗凝剂的生化和生物学潜力。该提议是 创新性是因为我提出了一种克服当前VTE治疗的局限性的新方法； ii） 它探讨了一种多学科方法来调查特定目标； iii）它引入了新技术 具有专有的结构和机械方面。该项目也很重要，因为它将：i）确定 2-3在VTE和出血的动物模型中，有效，特异性和变构FXIIIA抑制剂用于将来评估； ii）提供新的工具来更好地了解FXIIIA在凝血生理和病理中的作用； iii）调查 一种通过变构调节FXIIIA调节FXIIIA的替代方法，为转化抗凝剂铺平了道路。