Method of cell surface activation and inhibition
AssigneeWashington University
InventorGregory I. Goldberg
Life of this patent
Estimating the term…
Abstract
Disclosed are a peptide and its use in a method of screening test compounds as potential inhibitors of matrix metalloproteinases. The peptide consists of residues Trp 574 to Asp 656 in the TIMP-2 binding site of the C-terminal domain of gelatinase-A as shown by SEQ ID NO:19. The method comprises determining the inhibitory effect of a test compound in a competitive inhibition assay with said peptide in which a Ki/Kd=>1 is deemed an inhibitory effect of said test compound, and in which Ki is the inhibitor constant of said test compound and Kd is the dissociation constant of said peptide.
Claims2
A target area for screening test compounds as inhibitors of matrix metalloproteinases consisting of a peptide of residues Trp574 to Asp656 in the tissue inhibitor of 72 kDa type IV collagenase binding site of the C-terminal domain of gelatinase-A as shown in SEQ ID NO:19.
A method of screening a test compound for inhibition of matrix metalloproteineases comprising:
- a) contacting said test compound and said metalloproteinease, and
- b) determining the inhibitory effect of said test compound in a competitive inhibition assay in which displacement of 125I-labeled peptide bound to the tissue inhibitor of 72 kDa type IV collagenase by unlabeled test compound and unlabeled peptide is measured and the Ki/Kd ratio is determined;
- wherein Ki/Kd≧1 indicates inhibition, Ki is the inhibitory constant of said test compound, Kd is the dissociation constant of said peptide, and wherein said peptide consists of residues Trp574 to Asp656 in the tissue inhibitor of 72 kDa type IV collagenase binding site of the C-terminal domain of gelatinase A as shown in SEQ ID NO. 19.
Dependencies are inferred from the claim text.