Inhibitors of MLL-Menin Interaction

MLL-Menin 相互作用的抑制剂

• 批准号: 7290277
• 负责人: MICHAEL L CLEARY
• 金额: $ 19.69万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7290277
• 关键词: Acute leukemia; Adult; Affect; Affinity; Amino Acids; Basic Science; Binding; Biochemical; Biological Assay; Cellular Assay; Child; Chimeric Proteins; Chromosomal translocation; Clinic; Code; Complex; Consensus; Developmental Gene; Drug Design; Dysmyelopoietic Syndromes; Epigenetic Process; Fluorescence Polarization; Generations; Genetic Transcription; Infant; Knowledge; Label; Lead; Link; MLL gene; Measures; Mediating; Menin; Methodology; Modality; Molecular; Molecular Abnormality; Morbidity - disease rate; Mutate; Mutation; Myeloid-Lymphoid Leukemia Protein; Oncogene Proteins; Oncogenic; Pathogenesis; Patients; Peptides; Phenocopy; Proteins; Proto-Oncogenes; Reagent; Recombinants; Regulation; Role; Solutions; Specificity; Standards of Weights and Measures; Therapeutic Agents; Therapeutic Intervention; Treatment Protocols; Tumor Suppressor Genes; Tumor Suppressor Proteins; alpha-Thalassemia; base; chemotherapy; cofactor; design; gain of function; genetic analysis; high throughput screening; histone methyltransferase; inhibitor/antagonist; leukemia; leukemogenesis; metaplastic cell transformation; mortality; mutant; novel therapeutics; outcome forecast; response; small molecule; small molecule libraries; therapy outcome; tumorigenesis; young adult

DESCRIPTION (provided by applicant): The MLL (Mixed Lineage Leukemia) gene codes for a histone methyltransferase, which functions as a transcriptional epigenetic regulatory factor for diverse subordinate genes of developmental and oncogenic importance. It is mutated by chromosomal translocations that result in the creation of chimeric fusion proteins whose gain of function is critically involved in the pathogenesis of a poor prognosis subset of acute leukemia. Recently, MLL oncoproteins have been shown to associate with menin, a product of the MEN1 tumor suppressor gene. Genetic analyses revealed that menin is required for MLL-mediated transcription and transformation, thus demonstrating an unusual and unprecedented role for a tumor suppressor protein serving as an essential cofactor for an oncoprotein in cellular transformation. The MLL- menin interaction is critically dependent on a 5 amino acid high affinity menin-binding motif, which is retained in all oncogenic forms of MLL, and therefore constitutes a potential target for pharmacologic manipulation of MLL function and possible therapeutic intervention. The overall objectives of the studies proposed in this application are to establish methodologies to isolate small molecules that specifically inhibit the MLL-menin interaction. In the first specific aim, a fluorescence polarization assay will be developed that is capable of quantitatively measuring recombinant menin binding in solution to a fluorescently labeled peptide corresponding to the MLL consensus binding sequence. In the second specific aim, the MLL-menin blocking assay will be configured and/or optimized for high throughput screening. Biochemical and cellular assays will also be devised and optimized for establishing the specificity and efficacy of small molecule inhibitors of MLL-menin interaction. A preliminary screen through a small molecule library of 1,280 compounds (LOPAC, Sigma) in a quantitative high throughput format will help identify technical problems and confirm the feasibility of a larger scale high throughput screen. Compounds that are capable of specifically interfering with MLL-menin interaction will provide valuable reagents for studying the unusual functional cooperative roles of these proteins in transcriptional epigenetic regulation and oncogenesis, and provide lead compounds for potential rational drug design in poor prognosis leukemia. Acute leukemias are an important cause of morbidity and mortality, and their effects are particularly significant because they often affect children and young adults. Despite improvements in therapy and outcome in recent years, the prognosis for long-term survival in most leukemia patients remains poor (1-4). Thus, there is an urgent need for novel therapeutic modalities based on rational drug design targeting the molecular abnormalities that underlie leukemia pathogenesis.

描述（由申请人提供）：MLL（混合谱系白血病）基因编码组蛋白甲基转移酶，其作为具有发育和致癌重要性的多种从属基因的转录表观遗传调节因子。它通过染色体易位发生突变，导致嵌合融合蛋白的产生，其功能的获得与急性白血病预后不良亚型的发病机制密切相关。最近，MLL 癌蛋白已被证明与 menin（MEN1 肿瘤抑制基因的产物）相关。遗传分析表明，menin 是 MLL 介导的转录和转化所必需的，从而证明肿瘤抑制蛋白作为细胞转化中癌蛋白的重要辅助因子具有不寻常且前所未有的作用。 MLL-menin 相互作用严重依赖于 5 个氨基酸的高亲和力 menin 结合基序，该基序保留在 MLL 的所有致癌形式中，因此构成 MLL 功能的药理学操作和可能的治疗干预的潜在靶点。本申请中提出的研究的总体目标是建立分离特异性抑制 MLL-menin 相互作用的小分子的方法。在第一个具体目标中，将开发一种荧光偏振测定法，能够定量测量溶液中重组 menin 与对应于 MLL 共有结合序列的荧光标记肽的结合。在第二个具体目标中，MLL-menin 阻断测定将被配置和/或优化以用于高通量筛选。还将设计和优化生化和细胞测定，以确定 MLL-menin 相互作用的小分子抑制剂的特异性和功效。以定量高通量形式对包含 1,280 种化合物的小分子库（LOPAC、Sigma）进行初步筛选将有助于识别技术问题并确认更大规模高通量筛选的可行性。能够特异性干扰 MLL-menin 相互作用的化合物将为研究这些蛋白质在转录表观遗传调控和肿瘤发生中不寻常的功能协同作用提供有价值的试剂，并为不良预后白血病的潜在合理药物设计提供先导化合物。急性白血病是发病和死亡的重要原因，其影响尤为显着，因为它们经常影响儿童和年轻人。尽管近年来治疗和结果有所改善，但大多数白血病患者的长期生存预后仍然较差 (1-4)。因此，迫切需要基于针对白血病发病机制的分子异常的合理药物设计的新治疗方式。