The broad, long term objective of this proposal is to elucidate the role of each of three types of human collagenases in experimental rheumatoid arthritis and experimental angiogenesis. The three target enzymes are human skin fibroblast collagenase (most active against type III collagen), human skin fibroblast type IV collagenase, and human neutrophil collagenase (most active against type I collagen). These enzymes will be purified from cultured human skin fibroblasts (type III and type IV enzymes), and from purulent human sputum (type I enzyme). The 75 human skin fibroblast collagenase (type III) inhibitors developed in the last renewal of this proposal are tripeptide analogs the best of which have Ki's of 0.02 uM. These inhibitors will be screened against the other two pure enzymes. Amino acid substitutions will be made in these compounds in order to maximize specificity for each of the three collagenases. Variation of the amino acid side chain at the P2 position in a hexapeptide substrate has been shown to differentiate between fibroblast (type III) and tumor (type IV) collagenase. New specific peptide substrates for continuous spectrophotometric or fluorimetric assay of each collagenase will also be developed based on these same amino acid substitutions. Our best specific inhibitors will be tested as antiangiogenic agents against experimental angiogenesis in the rat cornea. Positive results (inhibition) would prove the role of the respective collagenase in this pathological condition and suggest the use of collagenase inhibitors as anti-angiogenic drugs.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL027368-12
Application #
3339149
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1980-12-01
Project End
1994-06-30
Budget Start
1992-07-01
Budget End
1993-06-30
Support Year
12
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Kentucky
Department
Type
Schools of Medicine
DUNS #
832127323
City
Lexington
State
KY
Country
United States
Zip Code
40506
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Kortylewicz, Z P; Galardy, R E (1990) Phosphoramidate peptide inhibitors of human skin fibroblast collagenase. J Med Chem 33:263-73
Kortylewicz, Z P; Galardy, R E (1989) Phthaloyl-glycylP-isoleucyl-tryptophan benzylamide is a potent inhibitor of human skin fibroblast collagenase with a Ki of 25 nM. J Enzyme Inhib 3:159-62

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