Synthesis, structure, and structure-activity relationships of divalent thrombin inhibitors containing an alpha-keto-amide transition-state mimetic.
Synthesis, structure, and structure-activity relationships of divalent thrombin inhibitors containing an alpha-keto-amide transition-state mimetic.
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含有α-酮酰胺过渡态模拟物的二价凝血酶抑制剂的合成、结构和构效关系。
DOI：
10.1002/pro.5560050303
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Ripka,WC
中科院分区：
文献类型：
--
作者：
Krishnan,R;Tulinsky,A;Vlasuk,GP;Pearson,D;Vallar,P;Bergum,P;Brunck,TK;Ripka,WC
描述了一类新的二价凝血酶抑制剂，其包含连接活性位点结合基团和与纤维蛋白原结合外位点结合的基团的α-酮酰胺过渡态模拟物。这一新类别中最有效的成员 CVS995 的 X 射线晶体结构显示出与其他二价凝血酶抑制剂的许多共同特征，并清楚地定义了 α-酮酰胺基团的过渡态样结合。抑制剂活性位点部分的结构显示了连接凝血酶和抑制剂的侧链和主链原子的水分子网络。使用体外凝血酶抑制测定来比较新型含过渡态二价凝血酶抑制剂的直接肽类似物。肽与其 α-酮酰胺对应物的结合常数之间没有直接相关性。最有效的 cv-酮酰胺抑制剂 CVS995，其 aKi= 1 pM，不对应于最有效的二价肽，并且相对于天然凝血酶抑制剂水蛭素的等效区域，在外部位点结合区域中包含单个氨基酸缺失。这些新型二价凝血酶抑制剂的活性位点、过渡态和外部位点结合区域的相互作用能量不是相加的。
A new class of divalent thrombin inhibitors is described that contains an α‐keto‐amide transition‐state mimetic linking an active site binding group and a group that binds to the fibrinogen‐binding exosite. The X‐ray crystallographic structure of the most potent member of this new class, CVS995, shows many features in common with other divalent thrombin inhibitors and clearly defines the transition‐state‐like binding of the α‐keto‐amide group. The structure of the active site part of the inhibitor shows a network of water molecules connecting both the side‐chain and backbone atoms of thrombin and the inhibitor. Direct peptide analogues of the new transition‐state‐containing divalent thrombin inhibitors were compared using in vitro assays of thrombin inhibition. There was no direct correlation between the binding constants of the peptides and their α‐keto‐amide counterparts. The most potent cv‐keto‐amide inhibitor, CVS995, with aKi= 1 pM, did not correspond to the most potent divalent peptide and contained a single amino acid deletion in the exosite binding region with respect to the equivalent region of the natural thrombin inhibitor hirudin. The interaction energies of the active site, transition state, and exosite binding regions of these new divalent thrombin inhibitors are not additive.