PEPTIDE INHIBITORS OF EXTRACELLULAR MATRIX PROTEASES

这项建议的广泛、长期目标是阐明 在实验性类风湿性关节炎中， 关节炎和实验性血管生成。 三种目标酶是 人皮肤成纤维细胞胶原酶（对III型最有活性 胶原）、人皮肤成纤维细胞IV型胶原酶和人胶原酶。 中性粒细胞胶原酶（对I型胶原最有活性）。 这些 将从培养的人皮肤成纤维细胞（III型）中纯化酶 和IV型酶），和来自脓性人痰（I型酶）。 75人皮肤成纤维细胞胶原酶（III型）抑制剂开发 在最近更新的这一建议是三肽类似物最好的 其Ki为0.02 μ M。 将对这些抑制剂进行筛选， 另外两种纯酶。 氨基酸取代将在 这些化合物，以最大限度地提高特异性的每一个三个 胶原酶 P2位氨基酸侧链的变异 在六肽底物中已经显示出区分 成纤维细胞（III型）和肿瘤（IV型）胶原酶。 新的特定 用于连续分光光度或荧光测定的肽底物 每种胶原酶的测定也将基于这些相同的结果来开发。 氨基酸取代。 我们最好的特异性抑制剂将作为抗血管生成剂进行测试 对抗大鼠角膜中的实验性血管生成。 积极成果 （抑制）将证明各自的胶原酶在这一过程中的作用。 病理条件，并建议使用胶原酶抑制剂， 抗血管生成药物

The broad, long term objective of this proposal is to elucidate the role of each of three types of human collagenases in experimental rheumatoid arthritis and experimental angiogenesis. The three target enzymes are human skin fibroblast collagenase (most active against type III collagen), human skin fibroblast type IV collagenase, and human neutrophil collagenase (most active against type I collagen). These enzymes will be purified from cultured human skin fibroblasts (type III and type IV enzymes), and from purulent human sputum (type I enzyme). The 75 human skin fibroblast collagenase (type III) inhibitors developed in the last renewal of this proposal are tripeptide analogs the best of which have Ki's of 0.02 uM. These inhibitors will be screened against the other two pure enzymes. Amino acid substitutions will be made in these compounds in order to maximize specificity for each of the three collagenases. Variation of the amino acid side chain at the P2 position in a hexapeptide substrate has been shown to differentiate between fibroblast (type III) and tumor (type IV) collagenase. New specific peptide substrates for continuous spectrophotometric or fluorimetric assay of each collagenase will also be developed based on these same amino acid substitutions. Our best specific inhibitors will be tested as antiangiogenic agents against experimental angiogenesis in the rat cornea. Positive results (inhibition) would prove the role of the respective collagenase in this pathological condition and suggest the use of collagenase inhibitors as anti-angiogenic drugs.