Phosphoramidate peptide inhibitors of human skin fibroblast collagenase.
Phosphoramidate peptide inhibitors of human skin fibroblast collagenase.
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DOI：
10.1021/jm00163a044
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发表时间：
1990
影响因子：
7.3
通讯作者：
Z. Kortylewicz;R. Galardy
中科院分区：
文献类型：
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作者：
Z. Kortylewicz;R. Galardy
合成了一系列广泛的 N-（单乙基磷酰基）肽，并检查了它们对纯化的人皮肤成纤维细胞胶原酶的抑制作用。在裂解位点 S1，所有报道的化合物都具有 (EtO)(OK)P(O) 基团，并且肽侧链向底物序列的 C 末端（直至 P5'）延伸。这些具有四面体杂化磷原子的氨基磷酸酯被认为是过渡态类似物抑制剂。它们对 Ki 值在微摩尔范围内的脊椎动物胶原酶表现出相当的抑制效力。其中最有效的是 (EtO)(OK)P(O)-Ile-TrpNHCH3 (68)，其抑制作用的 Ki 值为 1.5 microM，比 (EtO)(OK)P(O)-Ile-Ala-GlyOK (51)（Ki 为 140 microM）强近 100 倍，后者的序列与 P1' 中胶原蛋白 α 1 (I) 链的序列匹配，切割位点后的 P2'、P3'。制备了几种化合物，试图鉴定 S2'、S3' 和 S4' 结合位点的性质。 P2' 位置的丙氨酸被亮氨酸、苯丙氨酸、色氨酸或酪氨酸衍生物取代，导致 Ki 值与 51 相比明显较低，为 1.0-40 microM。在此位置没有发现大小上限或特异性，但在 P3' 位置（天然被甘氨酸残基占据）的类似替换产生较弱的抑制剂： (EtO)(OK)P(O)-Ile-Tyr(OBzl)-PheOK (57) 的 Ki 为 120 microM。六肽衍生物在270 microM-2 mM范围内活性较弱。所有抑制剂均通过合成硫肽内酯分光光度测定法进行评估。
An extensive series of N-(monoethylphosphoryl)peptides was synthesized and their inhibition of purified human skin fibroblast collagenase examined. At the cleavage site S1 all reported compounds have the (EtO)(OK)P(O) group and the peptide side chain extended toward the C-terminal end (up to P5') of the substrate sequence. These phosphoramidates with a tetrahedrally hybridized phosphorus atom are thought to be transition state analogue inhibitors. They exhibited fair inhibitory potency against this vertebrate collagenase having Ki values in the micromolar range. The most potent of these, (EtO)(OK)P(O)-Ile-TrpNHCH3 (68), inhibits with a Ki value of 1.5 microM and is nearly 100 times stronger than (EtO)(OK)P(O)-Ile-Ala-GlyOK (51) (Ki of 140 microM), which has the sequence matching that of the alpha 1 (I) chain of collagen in P1', P2', P3' after the cleavage site. Several compounds were prepared in an attempt to identify the nature of the S2', S3', and S4' binding sites. Alanine at the P2' position was replaced by leucine, phenylalanine, tryptophan, or tyrosine derivatives, resulting in Ki values in a significantly lower range, 1.0-40 microM, compared to 51. No upper size limitation or specificity has been found at this position, yet similar replacements at the P3' position, which is occupied naturally by a glycine residue, gave weaker inhibitors: (EtO)(OK)P(O)-Ile-Tyr(OBzl)-PheOK (57) had a Ki of 120 microM. Hexapeptide derivatives had weaker activities in the 270 microM-2 mM range. All inhibitors were evaluated by using the synthetic thio peptolide spectrophotometric assay.