Screening Metalloproteinase Inhibitors From Herbal Formulations

Matrix metalloproteinases (MMPs) are a class of zinc-requiringextracellular endopeptidases that together can degrade all components of theextracellular matrix and basement membranes. They play important roles inconnective tissue remodeling occurring in normal processes such asembryonic development, pregnancy, bone growth and wound heading.According to their substrate specificity and structure, MMP family can beclassified into subgroups: collagenases, stromelysins, gelatinases,membrane-type-MMPs, and other MMPs. The regulation of MMP activity isstrictly controlled at three different levels: gene transcription, proenzymeactivation, and enzyme activity. However, excessive or inappropriateexpression of MMPs may contribute to pathogenesis includinginflammation, cancer and cardiovascular diseases among others. Matrixmetalloproteinase inhibitors (MMPIs) are implicated as therapeutics agentsto treat these diseases. In order to screen MMPIs, we have chosen a numberof Chinese formulations used to treat inflammatory diseases, such astonsillitis, conjunctivitis, pharyngitis, nephritis and hepatitis. Here we reportKorean monkshood root, a stem of Aconitum coreanum (Levl.) Raipaics,can effectively inhibit MMPs.We have carried out these works:1. Determination of Matrix Metalloproteinase Activity Using QuenchedFluorescent Peptide Assay, determine Km and Kcat/Km of MMP-16 usingMichaelis-Menten Equation, determine activation energy of MMP-16 usingthe Arrhenius equation, determine the binding mechanism of GM6001inhibitors with MMP-16, and active site titration of enzyme concentrationusing Morrison equation.2. Korean monkshood root water solution was extracted with petroleumether, chloroform, acetoacetate and n-butyl alcohol. Using metalloproteinaseassays, determine inhibitory activities of different extracts on MMPs.Calculate IC50 of final water extract of Korean monkshood root towardMMPs. Investigate the effect of final water extract of Korean monkshoodroot on the viability of HT1080 cells.3. WebLab ViewerLite was used to display the active site ofproMMP-1, the active site of MT3-MMP, the structure of MMP-16 with thehydroxamic acid BB94, and compare the S1' pocket of cdMT3-MMP withcdMT1-MMP.