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

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

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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（例如心脏病发作和中风）的主要支柱并且也可以用于其他疾病，例如癌症和Covid-19。虽然几种抗凝剂诊所中使用了包括肝素，华法林和直接口服抗凝剂（DOAC）来自重大和小出血不良反应。截至目前，一种抑制凝结的安全抗凝剂尚未开发出出血风险。在NHLBI的R01机制下，我们早些时候发现人为因子Xia（HFXIA）的肝素结合exosite 2与其他同源凝血明显不同蛋白酶（例如HFIIA，HFXA等）。我们制定了一种依赖阴离子硫酸盐的新型设计策略和芳香环对HFXIA的外部2的高度选择性识别，从而抑制酶活动。我们开发了一种高度有前途的变构抑制剂，称为硫酸化的D-Chiro肌醇（SCI），它是分为四个步骤合成，显示HFXIA的100倍选择性；在Exosite 2中结合的血浆FXIA与即使酶的活性位点共价阻止，亲和力也为20-60 nm。并抑制体内动脉每只动物（〜1 mg/kg）以250 µg（〜1 mg/kg）的静脉血栓形成，而无需增强尾巴出血。 Sci是以剂量高达25 mg/大鼠的剂量耐受，表明治疗窗口约为100。 SCI是一个非常有前途的抗凝物;然而，它的药代动力学并不是最好的。使用基于肝素的药物的提示，我们假设优化硫酸盐基团在D-Chiro肌醇支架上的数量和位置，同时还筛选它的各种立体异构体将改善HFXIA亲和力，抑制效力，PK，药效学（PD）和在进行辅助研究之前，化学，制造和对照（CMC）特性。电流提案着重于研究20个合理设计的SCI类似物的库，目的是至少识别高级舞台前开发的首席代理。具体目的是1）合成，基于SCI结构的抗凝效率和20个假定因子XIA抑制剂的出血； 2）体内抗血栓效率，DMPK，体外和体内抑制剂的毒性； 3）扩展非GMP合成， CMC，剂量升级效率和PD研究以鉴定一个或两个铅分子。 SCI已被要求美国专利（＃9,758,459 B2，标题为“因子Xia的变构调节剂，为抗凝剂剂”）发明家之一。关于合成，体外和体内效率，体外和IN的定量里程碑体内毒性将用于指导从R61到R33期的过渡。另外，最好的1（或2）类似物从20个开始，SCI的SCI将被确定为最有前途的进一步研究的药物设计的SCI类似物。