How a 尾-d-Glucoside Side Chain Enhances Binding Affinity to Thrombin of Inhibitors Bearing 2-Chlorothiophene as P1 Moiety: Crystallography, Fragment Deconstruction Study

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• 作者：Benny D. Belviso ; Rocco Caliandro ; Modesto de Candia ; Giorgia Zaetta ; Gianfranco Lopopolo ; Francesca Incampo ; Mario Colucci ; Cosimo D. Altomare
• 刊名：Journal of Medicinal Chemistry
• 出版年：2014
• 出版时间：October 23, 2014
• 年：2014
• 卷：57
• 期：20
• 页码：8563-8575
• 全文大小：497K
• ISSN：1520-4804

文摘

The 尾-d-glucose-containing compound 3, bearing 2-chlorothiophene and 1-isopropylpiperidine moieties as binders of the S1 and S4 pockets, respectively, proved to be potent competitive inhibitor of factor Xa (fXa, K_i = 0.090 nM) and thrombin (fIIa, K_i = 100 nM). The potency of 3 increases, over the parent compound 1, against fIIa (110-fold), much more than against fXa (7-fold). Experimental deconstruction of 3 into smaller fragments revealed a binding cooperativity of the P3/P4 and propylene-linked 尾-d-glucose fragments, stronger in fIIa (15.5 kJ路mol^鈥?) than in fXa (2.8 kJ路mol^鈥?). The crystal structure of human fIIa in complex with 3 revealed a binding mode including a strong H-bond network between the glucose O1鈥? O3鈥? and O5鈥?and two critical residues, namely R221a and K224, belonging to the Na⁺-binding site which may allosterically perturb the specificity sites. The potential of 3 as antithrombotic agent was supported by its ability to inhibit thrombin generation and to stimulate fibrinolysis at submicromolar concentration.