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.

Public Health Relevance

The objective of this proposal is to identify existing small molecule compounds that can be used to control the function of an enzyme that plays a key role in skin diseases, tumor metastasis, and may play a role in Alzheimer?s disease. These molecules could shed light on the functions of this important enzyme family in health and disease, and provide insights that enable development of new therapeutics for cancer, skin diseases, and Alzheimer?s disease.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Small Research Grants (R03)
Project #
1R03DA034600-01
Application #
8328055
Study Section
Special Emphasis Panel (ZRG1-BST-F (50))
Program Officer
Singh, Hari
Project Start
2012-05-01
Project End
2014-04-30
Budget Start
2012-05-01
Budget End
2013-04-30
Support Year
1
Fiscal Year
2012
Total Cost
$38,187
Indirect Cost
$13,187
Name
University of California Santa Barbara
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
094878394
City
Santa Barbara
State
CA
Country
United States
Zip Code
93106