Lead identification and pre-clinical studies on allosteric inhibitors of coagulation factor XIa

凝血因子 XIa 变构抑制剂的先导化合物鉴定和临床前研究

基本信息

批准号：
10722847
负责人：
Umesh Ramanlal Desai
金额：
$ 54.34万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10722847
关键词：
Active Sites Adverse effects Affinity Animals Anticoagulant therapy Anticoagulants Anticoagulation Area Binding Biological Biotechnology COVID-19 Chemistry Clinic Clinical Clinical Research Coagulation Process Cues Disease Dose Drug Costs Drug Interactions Drug Kinetics Drug usage Elements Enoxaparin Enzyme Inhibition Enzymes Factor XIa Fibrinolytic Agents Goals Health Hemorrhage Heparin Heparin Binding Human In Vitro Incubators Inositol Institution Lead Leadership Legal patent Libraries Location Malignant Neoplasms Measures Minor Myocardial Infarction Names National Heart, Lung, and Blood Institute New Agents Oral Outpatients Pathway interactions Peptide Hydrolases Pharmaceutical Preparations Pharmacodynamics Phase Plasma Population Characteristics Positioning Attribute Preparation Property Rattus Renal clearance function Research Project Grants Risk Role Self Administration Stereoisomer Stroke Structure Sulfate Tail Technology Therapeutic Thrombosis Toxic effect Unstable angina Venous Thrombosis Warfarin analog cost design drug development drug metabolism drug standard enzyme activity experience improved in vivo inhibitor interest manufacture novel pharmacokinetics and pharmacodynamics preclinical development preclinical study rational design scaffold scale up screening small molecule thrombotic

项目摘要

Anticoagulants are the mainstay in the treatment of thrombotic diseases, such as heart attack and stroke, and may also be used in other diseases such as cancer and COVID-19. Although several anticoagulants including heparins, warfarin, and direct oral anticoagulants (DOACs) are used in the clinic, each agent suffers from major and minor bleeding adverse effects. As of now, a safe anticoagulant that inhibits coagulation without bleeding risk has not been developed as yet. Under the NHLBI's R01 mechanism, we earlier discovered that the heparin-binding exosite 2 of human factor XIa (hFXIa) is remarkably different from other homologous coagulation proteases (e.g., hFIIa, hFXa and others). We developed a novel design strategy that relies on anionic sulfates and aromatic rings to effect highly selective recognition of hFXIa's exosite 2 resulting in inhibition of enzyme activity. We developed a highly promising allosteric inhibitor, named sulfated D-chiro-inositol (SCI), which was synthesized in four steps, displayed >100-fold selectivity for hFXIa; bound plasma FXIa in exosite 2 with an affinity of 20–60 nM even when the enzyme's active site was covalently blocked; and inhibited in vivo arterial and venous thrombosis in the rat at 250 µg per animal (~1 mg/kg) without enhancing tail bleeding. SCI was tolerated at doses as high as 25 mg/rat suggesting a therapeutic window of ~100. SCI is a highly promising anticoagulant; yet its pharmacokinetics is not the best. Using cues from heparin-based drugs, we hypothesize that optimizing the number and position of sulfate groups on the D-chiro inositol scaffold, while also screening its various stereoisomers would improve hFXIa affinity, inhibition potency, PK, pharmacodynamics (PD), and chemistry, manufacturing and controls (CMC) properties before embarking on IND-enabling studies. The current proposal focuses on studying a library of 20 rationally designed SCI analogs with the goal of identifying at least one LEAD AGENT for advanced stage pre-clinical development. The specific aims are 1) synthesis, anticoagulation efficacy, and bleeding of 20 putative factor XIa inhibitors based on the SCI structure; 2) in vivo antithrombotic efficacy, DMPK, in vitro and in vivo toxicity of inhibitors; and 3) scaled-up non-GMP synthesis, CMC, dose escalation efficacy, and PD studies to identify one or two lead molecules. SCI has been claimed in a US patent (#9,758,459 B2 titled `Allosteric modulators of factor XIa as anticoagulant agents') with the PI as one of the inventors. Quantitative milestones with regard to synthesis, in vitro & in vivo efficacy, in vitro and in vivo toxicity will be used to guide the transition from the R61 to R33 Phase. Alternatively, the best 1 (or 2) analogs of SCI would be identified as the most promising agent for further IND-enabling studies starting from the 20 designed SCI analogs.

抗凝剂是治疗血栓性疾病的主要药物，例如心脏病和中风，尽管有多种抗凝剂，但也可用于其他疾病，例如癌症和 COVID-19。包括肝素、华法林、直接口服抗凝剂（DOAC）等在临床上使用，每种药物都受到影响迄今为止，一种安全的抗凝剂可以抑制凝血，而不会产生严重和轻微的出血不良反应。在NHLBI的R01机制下，我们之前还没有发现出血风险。人因子 XIa (hFXIa) 的肝素结合外位点 2 与其他同源凝血显着不同我们开发了一种依赖阴离子硫酸盐的新颖设计策略。和芳香环，以实现 hFXIa 外部位点 2 的高度选择性识别，从而抑制酶我们开发了一种非常有前途的变构抑制剂，称为硫酸化 D-手性肌醇 (SCI)，它是分四步合成，对 hFXIa 显示 >100 倍的选择性；在外部位点 2 中结合血浆 FXIa 即使酶的活性位点被共价阻断并抑制体内动脉，亲和力仍为 20–60 nM；每只动物服用 250 µg（约 1 mg/kg）时，大鼠出现静脉血栓，但不会增加尾部出血。高达 25 mg/大鼠的耐受剂量表明约 100 的 SCI 治疗窗是非常有前途的。抗凝剂；但根据基于肝素的药物的线索，它的药代动力学并不是最好的。优化D-手性肌醇支架上硫酸基的数量和位置，同时还筛选其各种立体异构体可提高 hFXIa 亲和力、抑制效力、PK、药效学 (PD) 和开始 IND 研究之前的化学、制造和控制 (CMC) 特性。该提案的重点是研究由 20 个合理设计的 SCI 类似物组成的文库，目标是至少识别高级临床前开发的一位主要代理具体目标是 1) 合成， 2) 体内 20 种假定的因子 XIa 抑制剂的抗凝功效和出血；抗血栓功效、DMPK、抑制剂的体外和体内毒性；以及 3) 扩大非 GMP 合成，已在 CMC、剂量递增疗效和 PD 研究中鉴定出一种或两种先导分子 SCI。一项美国专利（#9,758,459 B2，标题为“因子 XIa 的变构调节剂作为抗凝剂”），PI 为发明人之一，在合成、体外和体内功效方面的定量里程碑。体内毒性将用于指导从 R61 到 R33 阶段的过渡，或者，最好的 1（或 2）个类似物。 SCI 将被确定为从 20 世纪 20 年代开始进行进一步 IND 研究的最有前途的药物设计了 SCI 类似物。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Merging cultures and disciplines to create a drug discovery ecosystem at Virginia commonwealth university: Medicinal chemistry, structural biology, molecular and behavioral pharmacology and computational chemistry.

融合文化和学科，在弗吉尼亚联邦大学创建药物发现生态系统：药物化学、结构生物学、分子和行为药理学以及计算化学。

DOI：
发表时间：
2023-09
期刊：
影响因子：
0
作者：
Kellogg, Glen E;Cen, Yana;Dukat, Malgorzata;Ellis, Keith C;Guo, Youzhong;Li, Jiong;May, Aaron E;Safo, Martin K;Zhang, Shijun;Zhang, Yan;Desai, Umesh R
通讯作者：
Desai, Umesh R

Glycosaminoglycan microarrays for studying glycosaminoglycan-protein systems.

用于研究糖胺聚糖-蛋白质系统的糖胺聚糖微阵列。