INHIBITION OF EXTRACELLULAR MATRIX METALLOPROTEINASES

破坏细胞外基质，包括胶原蛋白 纤维构成牙周的骨骼框架 结缔组织，是炎症的主要后果 牙周病的危害 大量证据表明， 细胞外基质的这种降解是由 高度同源的蛋白酶家族，由 内源性和炎性细胞。 这些蛋白酶都 被认为是锌金属酶，它们共同 称为基质金属蛋白酶（MMP）。 五大MMP 包括成纤维细胞型胶原酶、明胶酶和溶基质素， 和明胶酶。 虽然 目前认为，这些MMP的集体行动是 负责牙周结缔组织的破坏 疾病，个别MMP在实际中的确切作用 细胞介导的过程尚未被很好地理解。 这种知识 只能从基质降解的研究中获得 活细胞产生的大分子，其中整个分解代谢 机器完好无损。 特定MMP的参与必须， 因此，可以通过其特异性抑制作用进行评估。 在这 研究特异性的有效抑制剂的每一个五个MMP将是 制备和它们在阻断基质分解中的功效 大分子（I、IV和V型胶原蛋白和明胶， 纤连蛋白、层粘连蛋白、蛋白聚糖核心蛋白） 将评估成纤维细胞和中性粒细胞。 这些实验 应该可以让我们确定每个MMP在细胞中的作用- 介导的每种生理底物的破坏。 设计 的抑制剂将基于肽的广泛研究， 五种MMP中每一种的底物特异性。 抑制剂将 是底物类似物，其中易断裂的肽键被替换， 与官能团（次膦酸酯，硫化物，亚砜，砜， 酮和氟酮）能够与活性物质相互作用 每个MMP的位点锌原子。 这一原则已被用于 以前开发血管紧张素转换酶抑制剂 其现在被广泛用于控制人类的高血压。 我们的数据表明，虽然高度同源，五个MMP 具有足够不同的肽底物特异性， 允许构建高度特异性的有效抑制剂， 五种酶的每一种。

Destruction of the extracellular matrix, including the collagen fibers which constitute the skeletal framework of periodontal connective tissues, is a major consequence of inflammatory periodontal disease in humans. A body of evidence suggests that this degradation of the extracellular matrix is catalyzed by a family of highly homologous proteinases that are released by endogenous and inflammatory cells. These proteinases are all believed to be zinc metalloenzymes and they are collectively referred to as matrix metalloproteinases (MMP). The five major MMP include fibroblast-type collagenase, gelatinase, and stromelysin, and neutrophil-type collagenase and gelatinase. While it is presently believed that the collective action of these MMP is responsible for the destruction of connective tissue in periodontal disease, the precise roles of the individual MMP in the actual cell-mediated process is not yet well understood. Such knowledge can only be obtained from the study of the degradation of matrix macromolecules by live cells in which the entire catabolic machinery is intact. The involvement of specific MMP must, therefore, be assessed through their specific inhibition. In this study specific potent inhibitors of each of the five MMP will be prepared and their efficacy in blocking the breakdown of matrix macromolecules (types I, IV and V collagens and gelatins, fibronectin, laminin, proteoglycan core protein) by live fibroblasts and neutrophils will be assessed. These experiments should allow us to establish the role of each MMP in the cell- mediated destruction of each physiological substrate. The design of the inhibitors will be based on extensive studies of the peptide substrate specificity of each of the five MMP. The inhibitors will be substrate analogs in which the scissile peptide bond is replaced with a functional group (phosphinate, sulfide, sulfoxide, sulfone, ketone and fluoroketone) capable of interacting with the active site zinc atom of each MMP. This principle has been used previously to develop inhibitors of angiotensin converting enzyme which are now being widely used to control hypertension in humans. Our data indicate that, although highly homologous, the five MMP have sufficiently different peptide substrate specificities to permit construction of potent inhibitors which are highly specific for each of the five enzymes.