Servicio de Hematología y Oncología Médica, H. U. Morales Meseguer, Centro Regional de Hemodonación, University of Murcia, Spain.
J Med Chem. 2012 Jul 26;55(14):6403-12. doi: 10.1021/jm300621j. Epub 2012 Jul 17.
The medical and socioeconomic relevance of thromboembolic disorders promotes an ongoing effort to develop new anticoagulants. Heparin is widely used as activator of antithrombin but incurs side effects. We screened a large database in silico to find alternative molecules and predicted d-myo-inositol 3,4,5,6-tetrakisphosphate (TMI) to strongly interact with antithrombin. Isothermal titration calorimetry confirmed a TMI affinity of 45 nM, higher than the heparin affinity (273 nM). Functional studies, fluorescence analysis, and citrullination experiments revealed that TMI induced a partial activation of antithrombin that facilitated the interaction with heparin and low affinity heparins. TMI improved antithrombin inhibitory function of plasma from homozygous patients with antithrombin deficiency with a heparin binding defect and also in a model with endothelial cells. Our in silico screen identified a new, non-polysaccharide scaffold able to interact with the heparin binding domain of antithrombin. The functional consequences of this interaction were experimentally characterized and suggest potential anticoagulant therapeutic applications.
血栓栓塞性疾病的医学和社会经济学意义促使人们不断努力开发新型抗凝剂。肝素被广泛用作抗凝血酶的激活剂，但会产生副作用。我们在计算机上筛选了一个大型数据库，以寻找替代分子，并预测 D-肌醇 3,4,5,6-四磷酸（TMI）与抗凝血酶强烈相互作用。等温滴定量热法证实 TMI 的亲和力为 45 nM，高于肝素的亲和力（273 nM）。功能研究、荧光分析和瓜氨酸化实验表明，TMI 诱导抗凝血酶部分激活，促进与肝素和低亲和力肝素的相互作用。TMI 改善了具有抗凝血酶缺陷和肝素结合缺陷的纯合子患者血浆以及内皮细胞模型中的抗凝血酶抑制功能。我们的计算机筛选鉴定出一种新的、非多糖支架，能够与抗凝血酶的肝素结合域相互作用。这种相互作用的功能后果已通过实验进行了表征，并提示具有潜在的抗凝治疗应用。
