Serine protease zymogen activation by small molecules

小分子激活丝氨酸蛋白酶酶原

基本信息

批准号：
8460829
负责人：
Patrick S Daugherty
金额：
$ 3.71万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8460829
关键词：
Alleles Alzheimer&apos s Disease Amines Amyloid beta-Protein Atopic Dermatitis Attention Behavior Biological Assay Biological Process Brain Cadherins Cancer Patient Cell Adhesion Molecules Cells Cerebrospinal Fluid Chemicals Cleaved cell Collection Cysteine Protease Development Dipeptides Disease Energy Transfer Enzyme Precursors Enzymes Esophagus Excision Exhibits Family Family member Gene Expression Glioblastoma Goals Health Hepatocyte Growth Factor Hippocampus (Brain)Human Individual Inflammatory Insulinase Kidney Kininogenase Late Onset Alzheimer Disease Light Malignant Neoplasms Malignant neoplasm of ovary Mean Survival Times Mediating N-terminal Neoplasm Metastasis Neprilysin Neurons Organ Pancreas Peptide Hydrolases Pharmaceutical Chemistry Play Prostate Prostate carcinoma Proteins Proteolytic Processing Psoriasis Rattus Reporting Role ST14 gene Serine Protease Site Skin Skin Neoplasms Sodium Chloride Staging Structure-Activity Relationship Substrate Specificity Synaptic plasticity Syndrome Toxic effect Transcript Trypsinogen Tumor Cell Invasion United States National Institutes of Health Urokinase Work amyloid precursor protein processing biological systems caspase-3 chymotrypsin high throughput screening human KLK15 protein improved insight matriptase migration novel novel therapeutics programs screening skin disorder small molecule small molecule libraries tool tumor tumor progression

项目摘要

The objective of this project is to identify small molecule probes that selectively activate a chymotrypsin-like serine protease zymogen. Although zymogens are widely processed by proteolytic removal of a propeptide, this mechanism of activation is difficult to control in biological systems. Consequently, the biological functions of many proteases remain obscure. Here we propose to screen the NIH small molecule library collection to identify small molecule probes that activate a chymotrypsin-like serine protease (CSP) zymogen, without the requirement for proteolytic processing of the proenzyme. A high-throughput screening assay for CSP activating compounds has been developed and optimized using resonance energy transfer. Compounds that exhibit high potency selectivity over related family members, and structurally similar proteases will be downselected in secondary screening assays. Non-proteolytic zymogen activation will be verified using orthogonal assays. The best hits downselected from secondary screens will be subjected to two to three rounds of medicinal chemistry to further improve their potency and selectivity. The chemical probes arising from this work will be applied to study how serine proteases can be activated, and to investigate the functions of CSP in tumor metastasis and synaptic plasticity.

该项目的目的是鉴定选择性激活胰凝乳蛋白酶样丝氨酸蛋白酶原的小分子探针。尽管酶原广泛通过蛋白水解去除前肽进行加工，但这种激活机制在生物系统中难以控制。因此，许多蛋白酶的生物学功能仍然不清楚。在这里，我们建议筛选 NIH 小分子文库集合，以鉴定可激活胰凝乳蛋白酶样丝氨酸蛋白酶 (CSP) 酶原的小分子探针，而不需要对酶原进行蛋白水解处理。使用共振能量转移开发并优化了 CSP 激活化合物的高通量筛选测定法。对相关家族成员和结构相似的蛋白酶表现出高效选择性的化合物将在二次筛选测定中被下调。非蛋白水解酶原激活将使用正交测定来验证。从二级筛选中筛选出的最佳命中将接受两到三轮药物化学，以进一步提高其效力和选择性。这项工作产生的化学探针将用于研究丝氨酸蛋白酶如何被激活，并研究CSP在肿瘤转移和突触可塑性中的功能。