Activators of Procaspase-7

Procaspase-7 激活剂

• 批准号: 7169310
• 负责人: JAMES A WELLS
• 金额: $ 19.19万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-20 至 2007-07-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7169310
• 关键词: NIH Roadmap Initiative tag; X ray crystallography; apoptosis; biological signal transduction; chemical registry /resource; cysteine endopeptidases; drug discovery /isolation; fluorescent dye /probe; high throughput technology; small molecule; stimulant /agonist

DESCRIPTION (provided by applicant): Our primary objective is to develop a high throughput screening assay focused on the discovery of novel small molecules that promote activation of procaspase-7. Programmed cell death, or apoptosis, is used by multicellular organisms to eradicate physiologically- divergent cells that threaten development, homeostasis and overall survival. Disruption of the apoptotic cycle can lead to a number of life-threatening human disorders including cancer, immunodeficiency, autoimmune and neurodegenerative diseases. Apoptosis is initiated by a variety of pathways which incorporate a family of cysteine proteases, known as caspases, that cleave specific proteins that drive cell death. Apoptotic caspases are divided into "initiators" and "effectors" with the initiator caspases -8 and -9 converging to activate effector caspases -3 and -7. Effector caspases -3 and -7 then execute the final steps of apoptosis by destroying actin, nuclear lamin and many anti-apoptotic regulatory proteins. Significantly, an allosteric cavity has recently been discovered at the homodimeric interface of these highly conserved caspases and small molecules targeted to this region have been shown to regulate caspase activity. Such compounds cause reversible rearrangements that mimic their procaspase antecedents. This discovery represents an exciting opportunity for the potential identification of small molecule agonists that promote activation of procaspases and, thus, stimulate cellular apoptosis independent of regulatory cleavage by upstream initiator caspases -8 and -9. The identification of lead compounds with drug-like properties that allosterically induce procaspase-7 activation would be a significant and novel advancement in cancer therapeutics. We aim to use HTS combinatorial library screening to identify lead compounds that target allosteric surface cavities of procaspase 7. Procaspase-7 will be expressed and purified and a high throughput procaspase-7 activation fluorescence assay will be developed in both 96-well and 384-well formats. Hits from the HTS screens will be analyzed for specificity, mode of binding and structure activity relationships and common structural features will be established. Structures of procaspase-7 in complex with promising molecules will be determined by x-ray crystallography. HTS hits will subsequently be optimized into drug leads. The identification of lead compounds with drug-like properties that allosterically induce procaspase-7 activation would be a significant and novel advancement in cancer therapeutics.

描述（由申请人提供）：我们的主要目标是开发一种高通量筛选测定法，重点是发现促进 procaspase-7 激活的新型小分子。多细胞生物利用程序性细胞死亡或细胞凋亡来根除威胁发育、稳态和总体生存的生理差异细胞。细胞凋亡周期的破坏可能导致许多危及生命的人类疾病，包括癌症、免疫缺陷、自身免疫和神经退行性疾病。细胞凋亡由多种途径引发，其中包含半胱氨酸蛋白酶家族（称为半胱天冬酶），可裂解驱动细胞死亡的特定蛋白质。凋亡半胱天冬酶分为“启动子”和“效应子”，启动子半胱天冬酶-8和-9汇聚以激活效应子半胱天冬酶-3和-7。然后，效应器半胱天冬酶 -3 和 -7 通过破坏肌动蛋白、核纤层蛋白和许多抗凋亡调节蛋白来执行细胞凋亡的最后步骤。值得注意的是，最近在这些高度保守的半胱天冬酶的同二聚体界面上发现了变构空腔，并且针对该区域的小分子已被证明可以调节半胱天冬酶活性。此类化合物引起可逆重排，模仿其前体天冬氨酸蛋白酶原。这一发现为潜在鉴定小分子激动剂提供了令人兴奋的机会，这些激动剂可促进半胱天冬酶原的激活，从而刺激细胞凋亡，而不受上游起始子半胱天冬酶-8和-9的调节性裂解的影响。鉴定具有药物样特性、变构诱导 proaspase-7 激活的先导化合物将是癌症治疗领域的一项重大且新颖的进展。我们的目标是使用 HTS 组合文库筛选来鉴定靶向 procaspase 7 变构表面空腔的先导化合物。Procaspase-7 将得到表达和纯化，并且将在 96 孔和 384 孔中开发高通量 procaspase-7 激活荧光测定法。良好的格式。将分析来自 HTS 筛选的命中的特异性、结合模式和结构活性关系，并建立共同的结构特征。与有希望的分子复合的 procaspase-7 的结构将通过 X 射线晶体学确定。 HTS 命中随后将被优化为先导药物。鉴定具有药物样特性、变构诱导 proaspase-7 激活的先导化合物将是癌症治疗领域的一项重大且新颖的进展。