Novel Screen for Targeted CLL Therapeutics

靶向 CLL 治疗的新型筛选

• 批准号: 7746678
• 负责人: Dale J Christensen
• 金额: $ 28.91万
• 依托单位: COGNOSCI, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-16 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7746678
• 关键词: 11q; Accounting; Affinity; Amino Acids; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Apolipoprotein E; Apoptosis; Apoptotic; B-Lymphocytes; Binding; Biological; Biological Assay; Biological Availability; Blast Phase; Blood; Cell Count; Cell Cycle Regulation; Cell Survival; Cells; Cholesterol Homeostasis; Chromosome abnormality; Chronic Lymphocytic Leukemia; Chronic Myeloid Leukemia; Clinical Trials; Coupled; Cytotoxic Chemotherapy; Data; Development; Dose; Environment; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Evaluation; Fluorescence; Fluorescence Resonance Energy Transfer; Gene Mutation; Genetic Transcription; Gleevec; Grant; Human; Imatinib; Immunoprecipitation; In Vitro; Inflammation; Inflammatory; Investigational New Drug Application; Lead; Leukocytes; Libraries; MAPK14 gene; MAPK8 gene; Mature B-Lymphocyte; Mediating; Methods; Molecular Weight; Normal Cell; Nuclear; Oncogenes; Oral; Orphan; PPP2R1B gene; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Phase; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Population; Preparation; Principal Investigator; Production; Property; Protein Binding; Protein Dephosphorylation; Protein phosphatase; Proteins; Relative (related person); Reporting; Safety; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Surface Immunoglobulins; Survival Rate; Symptoms; Testing; Therapeutic; Toxicology; Transgenic Mice; Transgenic Organisms; Tumor Suppressor Proteins; Validation; Western World; abstracting; adult leukemia; base; chemical synthesis; cytotoxic; cytotoxicity; design; high throughput screening; human IRAK1 protein; in vitro activity; in vivo; inhibitor/antagonist; inorganic phosphate; killings; leukemia; mimetics; miniaturize; mitogen-activated protein kinase p38; mouse model; neoplastic cell; new therapeutic target; novel; novel strategies; novel therapeutics; outcome forecast; overexpression; phosphatase inhibitor; pill; pre-clinical; prevent; pro-apoptotic protein; protein activation; protein phosphatase inhibitor-2; public health relevance; selective expression; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): Of the nearly 84,000 cases of leukemia in the Western world, B-cell chronic lymphocytic leukemia (B-CLL) is the most common and accounts for approximately 30% of all adult leukemia cases. It is characterized by the relentless accumulation of monoclonal mature B cells. Tumor cells from B-CLL patients show increased survival rates that have been shown to be due to inhibited apoptosis and alterations of genes involved in cell cycle control and cell survival. When evaluating the signaling pathways that are aberrantly activated in B-CLL cells, it was demonstrated that the kinase Akt is constitutively activated and the overstimulation of Akt results in activation of the NFkB pathway that leads to increased production of NO and inhibited apoptosis. Akt and the NFkB are downregulated by Protein Phosphatase-2a (PP2a). PP2a is a commonly known tumor suppressor that is inhibited by natural inhibitors. Recently, we have shown that the protein SET, a natural inhibitor of PP2a is overexpressed in B-CLL, leading to decreased PP2a activity in B-CLL cells. Reduced PP2a activity would be expected to lead to aberrant signaling through Akt; therefore, pharmacological activation of PP2a by inhibition of SET may provide a novel approach to development of B-CLL therapeutics. Cognosci has developed novel therapeutic peptides with potent anti-inflammatory activity in vitro and in vivo. In mechanistic studies we recently demonstrated that these peptides suppress phosphorylation and the accompanying activation of important inflammatory signaling by acting as antagonists of SET that activate SET. During a preliminary evaluation of several COG compounds in primary CD19+/CD5+ B-CLL cells from patients, we discovered that one of these compounds, (COG112) is highly and preferentially cytotoxic for human CLL cells with an EC50 of 224 nM and an EC50 for normal B-cells of >20 uM. Furthermore, this compound reduced white cell counts in an animal model of CLL to validate SET antagonism as a potential therapeutic approach for CLL. We now propose to develop a peptide displacement assay that can be used to screen compound libraries to identify small molecule SET antagonists and screen a diverse 30,000 compound library. PUBLIC HEALTH RELEVANCE: Of the nearly 84,000 cases of leukemia in the Western world, B-cell chronic lymphocytic leukemia (B-CLL) is the most common and accounts for approximately 30% of all adult leukemia cases. It is characterized by the relentless accumulation of monoclonal mature B cells. Tumor cells from B-CLL patients show increased survival rates that have been shown to be due to inhibited apoptosis and alterations of genes involved in cell cycle control and cell survival. When evaluating the signaling pathways that are aberrantly activated in B-CLL cells, it was demonstrated that the kinase Akt is constitutively activated and the overstimulation of Akt results in activation of the NFkB pathway that leads to increased production of NO and inhibited apoptosis. Akt and the NFkB are downregulated by Protein Phosphatase-2a (PP2a). PP2a is a commonly known tumor suppressor that is inhibited by natural inhibitors. Recently, we have shown that the protein SET, a natural inhibitor of PP2a is overexpressed in B-CLL, leading to decreased PP2a activity in B-CLL cells. Reduced PP2a activity would be expected to lead to aberrant signaling through Akt; therefore, pharmacological activation of PP2a by inhibition of SET may provide a novel approach to development of B-CLL therapeutics. Cognosci has developed novel therapeutic peptides with potent anti-inflammatory activity in vitro and in vivo. In mechanistic studies we recently demonstrated that these peptides suppress phosphorylation and the accompanying activation of important inflammatory signaling by acting as antagonists of SET that activate SET. During a preliminary evaluation of several COG compounds in primary CD19+/CD5+ B-CLL cells from patients, we discovered that one of these compounds, (COG112) is highly and preferentially cytotoxic for human CLL cells with an EC50 of 224 nM and an EC50 for normal B-cells of >20 uM. Furthermore, this compound reduced white cell counts in an animal model of CLL to validate SET antagonism as a potential therapeutic approach for CLL. We now propose to develop a peptide displacement assay that can be used to screen compound libraries to identify small molecule SET antagonists and screen a diverse 30,000 compound library.

描述(申请人提供)：在西方世界近84,000例白血病中，B细胞慢性淋巴细胞白血病(B-CLL)是最常见的，约占所有成人白血病病例的30%。它的特点是单克隆性成熟B细胞的持续积累。来自B-CLL患者的肿瘤细胞显示出更高的存活率，这被证明是由于细胞周期控制和细胞生存的基因被抑制和改变所致。在评估B-CLL细胞中异常激活的信号通路时，研究表明，Akt是结构性激活的，而Akt的过度刺激导致NFkB通路的激活，从而导致NO的产生增加，并抑制细胞凋亡。蛋白磷酸酶-2a(PP2A)下调AKT和NFkB的表达。PP2A是一种广为人知的肿瘤抑制因子，可被天然抑制剂抑制。最近，我们发现PP2A的天然抑制剂蛋白SET在B-CLL中高表达，导致B-CLL细胞中PP2A活性降低。PP2A活性降低可能通过Akt导致异常信号转导，因此，通过抑制SET来药理激活PP2A可能为B-CLL药物的开发提供新的途径。Cognosci已经开发出在体外和体内具有强大抗炎活性的新型治疗性多肽。在机制研究中，我们最近证实这些多肽通过作为激活SET的SET的拮抗剂来抑制磷酸化和伴随的重要炎症信号的激活。在对患者原代CD19+/CD5+B-CLL细胞中的几种COG化合物进行初步评估时，我们发现其中一种化合物(COG112)对人CLL细胞具有高度和优先的细胞毒性，其EC50为224 nM，对正常B细胞的EC50为20微米。此外，在CLL的动物模型中，该化合物降低了白细胞计数，以验证SET拮抗作为CLL的潜在治疗方法。我们现在建议开发一种多肽置换试验，可以用于筛选化合物文库，以识别小分子集拮抗剂，并筛选不同的30,000个化合物文库。公共卫生相关性：在西方世界近84,000例白血病病例中，B细胞慢性淋巴细胞白血病(B-CLL)是最常见的，约占所有成人白血病病例的30%。它的特点是单克隆性成熟B细胞的持续积累。来自B-CLL患者的肿瘤细胞显示出更高的存活率，这被证明是由于细胞周期控制和细胞生存的基因被抑制和改变所致。在评估B-CLL细胞中异常激活的信号通路时，研究表明，Akt是结构性激活的，而Akt的过度刺激导致NFkB通路的激活，从而导致NO的产生增加，并抑制细胞凋亡。蛋白磷酸酶-2a(PP2A)下调AKT和NFkB的表达。PP2A是一种广为人知的肿瘤抑制因子，可被天然抑制剂抑制。最近，我们发现PP2A的天然抑制剂蛋白SET在B-CLL中高表达，导致B-CLL细胞中PP2A活性降低。PP2A活性降低可能通过Akt导致异常信号转导，因此，通过抑制SET来药理激活PP2A可能为B-CLL药物的开发提供新的途径。Cognosci已经开发出在体外和体内具有强大抗炎活性的新型治疗性多肽。在机制研究中，我们最近证实这些多肽通过作为激活SET的SET的拮抗剂来抑制磷酸化和伴随的重要炎症信号的激活。在对患者原代CD19+/CD5+B-CLL细胞中的几种COG化合物进行初步评估时，我们发现其中一种化合物(COG112)对人CLL细胞具有高度和优先的细胞毒性，其EC50为224 nM，对正常B细胞的EC50为20微米。此外，在CLL的动物模型中，该化合物降低了白细胞计数，以验证SET拮抗作为CLL的潜在治疗方法。我们现在建议开发一种多肽置换试验，可以用于筛选化合物文库，以识别小分子集拮抗剂，并筛选不同的30,000个化合物文库。