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 血栓形成是常见的并发症，常发生在内脏静脉和动脉等不常见部位， 慢性骨髓增生性肿瘤（MPN）患者死亡和发病的主要原因。 对MPN血栓形成的有效治疗方法的研究刚刚起步。当前慢性前列腺炎的一线治疗 相MPN包括阿司匹林、羟基脲、干扰素α或阿那格雷。然而，这些治疗方法 仍然是次优的，具有持续的血栓形成、出血和生活质量受损的风险。针对性 在MPN的这一阶段也缺乏用于血栓形成的分子治疗。Aplexis，Inc是一家初创公司， 专注于开发治疗MPN慢性期血栓形成的新方法，特别是 靶向在这些疾病中通常被激活的JAK 2途径的下游效应物。之一 这些效应物是Pleckstrin-2（Plek 2），其是JAK 2-STAT 5途径的新靶点。先前的研究 Plek 2的缺失改善了JAK 2 V617 F突变体诱导的骨髓增殖表型， JAK 2 V617 F MPN小鼠模型的血栓形成和致死性。重要的是，Plek 2在骨骼中过度表达， 来自JAK 2 V617 F阳性慢性MPN患者的骨髓和外周血单核细胞。鉴于 Plek 2在MPN血栓形成发病机制中的重要性，西北大学Ji实验室 使用基于计算机的高通量筛选鉴定Plek 2小分子抑制剂的命中化合物 和基于细胞的测定。初步数据显示，命中化合物显着抑制Plek 2的功能， 成红细胞增殖和板状伪足形成。受欢迎的化合物也会大大减少 骨髓增生和血栓形成。初步的机制研究表明， Plek 2作为支架蛋白发挥功能以募集PI 3 K效应蛋白并增强PI 3 K-Akt信号传导。Plek2 抑制剂与Plek 2结合并破坏这种募集，从而阻断PI 3 K-Akt信号传导并抑制细胞增殖。 过度增殖总之，这些发现使我们假设Plek 2抑制剂阻断细胞增殖， 在体外和体内过度增殖和预防MPN中血栓形成。第一阶段SBIR项目的目标是 建立Plek 2抑制剂在MPN血栓形成中的治疗效果的原理证据。在 目的1，我们提出开发具有Plek 2诱导的细胞增殖的有效抑制作用的先导化合物 在体外使用药物化学和基于细胞的测定。在目标2中，我们将检查疗效， 在具有血栓形成的临床前MPN小鼠模型中的先导化合物的药代动力学和毒性。阿朴西斯 西北大学创新和新企业办公室（INVO）的大力支持， 该技术的相关专利申请知识产权的独家许可。协作 与西北大学Ji实验室的合作将确保拟议研究的成功， 将Aplexis定位于Plek 2抑制剂和pre-IND临床候选药物生产的下一步 第二阶段调查。