Type Iv Collagenase Inhibitor Screening And Activity

Matrix metalloproteinases (MMPs) has been viewed as a promising target in tumor therapy due to their key role in tumor growth, angiogenesis, invasion and metastasis. Type IV coUagenase (Col. IV) is an important member of MMPs family which can destroy the integrity of basement membrane and extracellular matrix through the degrading of type IV collagen, thus facilitate the process of tumor invasion and metastasis. Also, the expression and secretion of type IV coUagenase is highly elevated in tumor cells and adjacent interstitial tissue, especially in malignancy with high potential of invasion and metastasis. Studies using type IV coUagenase as a molecular target have achieved a lot in tumor therapy especially in the control of tumor metastasis. In this study we tried to find collagenase-inhibitors in synthetic compounds and natural products by 2 drug screening models. We succeeded to get 2 active polysaccharides from microbe broths. And we also evaluated antitumor effects and anti-invasion effects of H25, a quinolone compound found through screening.1. Application of an indirect-ELISA screening model for Col. IV inhibitor and establishment of a novel screening model byfluorescence substrate assay (FSA).Ab-3D6 showed both the binding and neutralizing activity to type IV coUagenase, so a hypothesis was brought forward that those compounds that can inhibit the binding of 3D6 to type IV coUagenase or PG cells may affect the activity of type IV coUagenase. Based on the hypothesis, an indirect-ELISA model was established for Col. IV inhibitors. Then we gave a doubt about the validity of the model after we practised it, for it doesn't agree with gelatinase-zymogram assay. So we tried to establish another model for screening inhibitors and we choosed the fluorescence substrate assay. MMPs can cleave this peptide between Gly-Leu, and fluorescence at 346nm by tryptophan occurs once cleavage separates it from the...