INHIBITION OF EXTRACELLULAR MATRIX METALLOPROTEINASES

破坏细胞外基质，包括胶原 构成牙周骨架的纤维 结缔组织，是炎症的主要后果 人类的牙周病。大量证据表明 这种细胞外基质的降解是由一种 高度同源的蛋白水解酶家族，由 内源性和炎性细胞。这些水解酶都是 被认为是锌金属酶，它们共同 称为基质金属蛋白酶(MMPs)。五大基质金属氧化物 包括成纤维细胞型胶原酶、明胶酶和基质分解酶， 中性粒细胞型胶原酶和明胶酶。虽然它是 目前认为这些基质金属蛋白酶的集体行动是 对牙周结缔组织的破坏负责 疾病，个体基质金属蛋白酶在实际中的确切作用 细胞介导的过程还不是很清楚。这样的知识 只有通过研究基质的降解才能得到 大分子由活细胞在其中整个分解代谢 机器完好无损。特定的基质金属蛋白酶的参与必须， 因此，要通过对它们的特异性抑制来评估。在这 研究五种基质金属蛋白酶的特定有效抑制剂 它们的制备及其阻断基质分解的效果 大分子(I、IV和V型胶原蛋白和明胶， 纤维连接蛋白、层粘连蛋白、蛋白多糖核心蛋白) 将对成纤维细胞和中性粒细胞进行评估。这些实验 应该能让我们确定每种基质金属蛋白酶在细胞中的作用- 介导对每种生理底物的破坏。设计 抑制剂的选择将基于对多肽的广泛研究 5种基质金属蛋白酶的底物特异性。抑制剂将 BE底物类似物，其中可剪断的多肽键被取代 带有官能团(亚磷酸酯、硫化物、亚硫醚、砜、 酮和氟酮)能够与活性物质相互作用 每种基质金属丝的锌原子位置。这一原则已被用于 之前开发血管紧张素转换酶抑制剂 它现在被广泛用于控制人类的高血压。 我们的数据表明，尽管同源性很高，但五种基质金属蛋白酶 具有完全不同的多肽底物特异性 允许构建高度特异的有效抑制剂 五种酶中的每一种。

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.