Developing small molecule inhibitors of Pleckstrin-2 to treat thrombosis

开发 Pleckstrin-2 小分子抑制剂来治疗血栓形成

• 批准号: 10545992
• 负责人: Jing Yang
• 金额: $ 39.72万
• 依托单位: APLEXIS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545992
• 关键词: Anemia; Arteries; Aspirin; Binding; Binding Proteins; Biological Assay; Biometry; Blood Cell Count; Blood Vessels; Bone Marrow; Cell Proliferation; Cells; Chemistry; Chronic; Chronic Phase; Clinical Management; Clinical Trials; Collaborations; Complication; Data; Development; Disease; Drug Kinetics; Ensure; Erythroblasts; Erythropoietin; FDA approved; Foundations; Future; Generations; Genetic study; Goals; Hematopoiesis; Hemorrhage; Hemorrhagic Thrombocythemia; Impairment; In Vitro; Injections; Intellectual Property; Interferon-alpha; Investigation; JAK2 gene; Knock-in; Knock-in Mouse; Knock-out; Laboratories; Lead; Legal patent; Letters; Licensing; Mediating; Membrane; Modeling; Morbidity - disease rate; Mus; Mutation; Myeloproliferation; Myeloproliferative disease; Names; Neutrophilia; Pathogenesis; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Chemistry; Phase; Phenotype; Phosphatidylinositols; Polycythemia Vera; Positioning Attribute; Primary Myelofibrosis; Production; Proteins; Quality of life; Research; Reticulocytosis; Risk; Role; Scaffolding Protein; Signal Transduction; Site; Small Business Innovation Research Grant; Spleen; Splenomegaly; Stat5 protein; Technology; Testing; Therapeutic Effect; Thrombocytopenia; Thrombosis; Toxic effect; Universities; Veins; base; clinical candidate; effective therapy; efficacy evaluation; experimental study; high throughput screening; hydroxyurea; improved; in silico; in vivo; inhibitor; innovation; molecular targeted therapies; mortality; mouse genetics; mouse model; mutant; new therapeutic target; novel; novel strategies; novel therapeutic intervention; overexpression; pharmacokinetics and pharmacodynamics; platelet protein P47; pre-clinical; prevent; recruit; side effect; small molecule; small molecule inhibitor; success; thrombocytosis

PROJECT SUMMARY Thrombosis, often at unusual sites such as splanchnic vein and arteries, is common complication and one of the leading causes of mortality and morbidity in patients with chronic myeloproliferative neoplasms (MPNs). Developing effective therapies for thrombosis in MPNs is in its nascence. Current first-line therapy of chronic phase MPNs includes aspirin, hydroxyurea, interferon a, or anagrelide. However, these treatment approaches remain suboptimal with ongoing risks for thrombosis, hemorrhage, and impaired quality of life. Targeted molecular therapy at this stage of MPNs for thrombosis is also lacking. Aplexis, Inc is a startup company focusing on the development of new approaches to treat thrombosis in the chronic phase of MPNs, especially targeting the downstream effectors of the JAK2 pathway that is commonly activated in these disorders. One of these effectors is Pleckstrin-2 (Plek2) that is a novel target of the JAK2-STAT5 pathway. Previous studies have shown that loss of Plek2 ameliorated JAK2V617F mutant-induced myeloproliferative phenotypes, and reverted thrombosis and lethality of the JAK2V617F MPN mouse model. Importantly, Plek2 is overexpressed in the bone marrow and peripheral blood mononuclear cells from JAK2V617F positive chronic MPN patients. Given the significance of Plek2 in thrombosis pathogenesis in MPNs, the Ji laboratory at Northwestern University has identified hit compounds of Plek2 small molecule inhibitors using in silico-based high-throughput screenings and cell-based assays. Preliminary data show that the hit compounds significantly inhibit Plek2’s functions on erythroblast proliferation and lamellipodia formation in vitro. The hit compounds also dramatically reduce myeloproliferation and thrombosis in vivo in MPN mouse models. Preliminary mechanistic studies reveal that Plek2 functions as a scaffold protein to recruit PI3K effector proteins and enhances PI3K-Akt signaling. Plek2 inhibitors bind to Plek2 and disrupt this recruitment, which blocks PI3K-Akt signaling and inhibits cell hyperproliferation. Together, these findings lead us to hypothesize that Plek2 inhibitors block cell hyperproliferation in vitro and in vivo and prevent thrombosis in MPNs. The goal of this Phase I SBIR project is to establish proof-of-principle evidence for the therapeutic effects of Plek2 inhibitors in thrombosis in MPNs. In Aim 1, we propose to develop lead compounds with potent inhibitory effects of Plek2-induced cell proliferation in vitro using medicinal chemistry and cell-based assays. In Aim 2, we will examine the efficacy, pharmacokinetics, and toxicity of lead compounds in pre-clinical MPN mouse models with thrombosis. Aplexis has a strong support from Northwestern University’s Innovation and New Ventures Office (INVO) on an exclusive license for the associated patent-pending intellectual property from this technology. The collaboration with the Ji laboratory at Northwestern University will ensure the success of the proposed research, which will position Aplexis to the next step in the production of clinical candidate of Plek2 inhibitors and pre-IND investigation in Phase II.

项目摘要 血栓形成通常在方案静脉和动脉等异常部位，是常见的并发症，是 慢性骨髓增生性肿瘤（MPN）患者死亡率和发病率的主要原因。 在MPN中开发有效的血栓形成疗法是其无效的。慢性的当前一线治疗 相MPN包括阿司匹林，羟基脲，干扰素A或Anagrelide。但是，这些治疗方法 保持次优，以及血栓形成，出血和生活质量受损的持续风险。目标 在此阶段，MPNS的分子疗法也缺乏血栓形成。 Adlexis，Inc是一家创业公司 专注于在MPN的慢性阶段的新方法的发展，尤其是 针对通常在这些疾病中激活的JAK2途径的下游效应。之一 这些效果是PLECKSTRIN-2（PLEK2），它是JAK2-STAT5途径的新目标。先前的研究已有 表明PLEK2的损失可以改善JAK2V617F突变体诱导的脊髓增生性表型，并恢复 JAK2V617F MPN小鼠模型的血栓形成和致死性。重要的是，Plek2在骨头中过表达 来自JAK2V617F阳性慢性MPN患者的骨髓和外周血单核细胞。鉴于 PLEK2在MPN中的血栓形成发病机理中的重要性，西北大学的JI实验室具有 在基于硅的高通量筛选中使用的PLEK2小分子抑制剂的命中率化合物 和基于细胞的测定。初步数据表明，HIT化合物显着抑制PLEK2的功能 在体外，红细胞增殖和层状形成。命中化合物也大大减少 MPN小鼠模型中体内的骨髓增生和血栓形成。初步机械研究表明 PLEK2充当支架蛋白，可募集PI3K效应蛋白并增强PI3K-AKT信号传导。 plek2 抑制剂与plek2结合并破坏这种募集，从而阻断PI3K-AKT信号并抑制细胞 过度增殖。这些发现在一起，使我们假设plek2抑制剂阻止细胞 体外和体内过度增殖，并防止MPN中的血栓形成。这个阶段I SBIR项目的目标是 为了建立PLEK2抑制剂在MPN中血栓形成中的治疗作用的原则证据。 AIM 1，我们建议开发具有PLEK2诱导细胞增殖潜在抑制作用的铅化合物 使用药物化学和基于细胞的测定法进行体外。在AIM 2中，我们将检查效率， 具有血栓形成前临床前MPN小鼠模型中的药代动力学和铅化合物的毒性。脚扶脚 在西北大学的创新和新风险办公室（Invo）方面得到了大力支持 该技术的相关专利知识产权的独家许可。合作 在西北大学的JI实验室中，将确保拟议研究的成功，这将 将氨脚定位为生产PLEK2抑制剂临床候选者的下一步 在第二阶段的研究。