Allosteric Modulators of Src-family Kinases for Acute Myeloid Leukemia

急性髓系白血病 Src 家族激酶的变构调节剂

• 批准号: 10475633
• 负责人: Ari Selzer
• 金额: $ 3.08万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10475633
• 关键词: 3-Dimensional; Active Sites; Acute Myelocytic Leukemia; Adult; Apoptosis; Bcr-Abl tyrosine kinase; Binding; Binding Sites; Biological Assay; Cell Line; Cell Proliferation; Cell Survival; Cells; Chronic Myeloid Leukemia; Clinic; Clinical; Complex; Crystallization; Data; Development; Diamines; Disease; Docking; Drug Targeting; FLT3 gene; Family member; Fellowship; Future; Gleevec; Growth; Hematopoietic; Hematopoietic Neoplasms; Imatinib; In Vitro; Investigation; Lead; Length; Link; Malignant Neoplasms; Mammals; Mitosis; Modeling; Molecular Conformation; Mutation; Myelogenous; N-terminal; Nucleic Acid Regulatory Sequences; Patients; Pharmaceutical Chemistry; Pharmacology; Pharmacotherapy; Phosphotransferases; Point Mutation; Predisposition; Prognosis; Property; Protein Tyrosine Kinase; Pyrimidine; Receptor Protein-Tyrosine Kinases; Recombinants; Regulation; Resistance; Roentgen Rays; Role; SH3 Domains; Sampling; Severities; Signal Pathway; Signal Transduction; Site; Specificity; Structure; Surface Plasmon Resonance; Survival Rate; Testing; Therapeutic; Tissues; Treatment Efficacy; Treatment Failure; Tyrosine Kinase Inhibitor; Work; X-Ray Crystallography; acute myeloid leukemia cell; base; cancer initiation; cell growth; cell motility; experimental study; high throughput screening; improved; in silico; in vitro testing; in vivo; inhibitor; insight; kinase inhibitor; mRNA Differential Displays; member; mouse model; mutant; novel; novel drug class; novel strategies; overexpression; progenitor; resistance mutation; response; side effect; small molecule; src Homology Region 2 Domain; src-Family Kinases; synergism; therapeutic target; tumor progression; virtual

Abstract The eight mammalian Src-family members are non-receptor protein-tyrosine kinases which are involved in nearly all cell signaling pathways. Hck and Fgr are members of this family expressed almost exclusively in myeloid hematopoietic cells and their progenitors. The overexpression of these kinases has been linked to the development of, and a poorer prognosis in, acute myeloid leukemia (AML). AML is a common form of blood cancer in adults, with nearly 20,000 new cases per year in the US. About one-third of AML cases have activating mutations in the Flt3 receptor tyrosine kinase, including point mutations and internal tandem duplications. Current treatments for this subset of AML patients include ATP-site kinase inhibitors, although acquired resistance mutations commonly develop within one year of the start of treatment. Inhibitors for Hck and Fgr are also emerging as a new approach to AML therapy. Our group recently identified small molecules that bind to the regulatory domains of Hck (unique-SH3-SH2-linker) as opposed to the ATP-binding site of the kinase domain. Binding data and docking models suggest that these compounds bind to a site that requires a specific 3D- conformation of the SH3 and SH2 domains. They also decrease the viability of an AML cell line that overexpresses active Hck and Fgr. Based on these results, we hypothesize that these compounds suppress AML cell growth by interfering with downstream signaling via the SH3 and SH2 domains. In addition, the compounds may allosterically influence the conformation of the active site to favor ATP-site inhibitor action. We aim to expand upon these findings using in vitro kinase and binding assays to explore selectivity within the Src- kinase family and possible synergy with ATP-site inhibitors. The anti-AML mechanism of action of the compounds will be tested in AML cell line models by correlating their effects on kinase activity and downstream substrate activation with growth suppression and apoptosis. Finally, we will determine the binding site for these putative allosteric inhibitors within Hck by X-ray crystallography. These experiments will provide crucial insight for the future development of these novel compounds as a treatment for AML, either as stand-alone therapy or in combination with existing ATP-site inhibitors. 1

抽象的 八个哺乳动物 Src 家族成员是非受体蛋白酪氨酸激酶，它们参与几乎 所有细胞信号通路。 Hck 和 Fgr 是该家族的成员，几乎仅在骨髓中表达 造血细胞及其祖细胞。这些激酶的过度表达与 急性髓系白血病（AML）的发展和较差的预后。 AML 是一种常见的血液形式 成人癌症，美国每年新增病例近 20,000 例。大约三分之一的 AML 病例已激活 Flt3 受体酪氨酸激酶突变，包括点突变和内部串联重复。当前的 针对这部分 AML 患者的治疗包括 ATP 位点激酶抑制剂，尽管获得性耐药 突变通常在治疗开始后一年内发生。 Hck 和 Fgr 抑制剂也 正在成为 AML 治疗的新方法。我们的小组最近发现了与 Hck 的调节域（独特的 SH3-SH2 连接子）与激酶域的 ATP 结合位点相反。 结合数据和对接模型表明这些化合物结合到需要特定 3D- SH3 和 SH2 结构域的构象。它们还会降低 AML 细胞系的活力 过表达活跃的 Hck 和 Fgr。基于这些结果，我们假设这些化合物抑制 通过 SH3 和 SH2 结构域干扰下游信号传导来促进 AML 细胞生长。此外， 化合物可能会变构地影响活性位点的构象，以有利于 ATP 位点抑制剂的作用。我们 旨在使用体外激酶和结合测定来扩展这些发现，以探索 Src- 内的选择性 激酶家族以及与 ATP 位点抑制剂可能的协同作用。抗AML作用机制 化合物将在 AML 细胞系模型中进行测试，通过关联它们对激酶活性和下游的影响 底物激活与生长抑制和细胞凋亡。最后，我们将确定这些的结合位点 通过 X 射线晶体学确定 Hck 内推定的变构抑制剂。这些实验将提供重要的见解 未来开发这些新化合物作为 AML 的治疗方法，无论是作为独立疗法还是 与现有的 ATP 位点抑制剂组合。 1