The Computer Virtual Design And Pharmacological Activity Research Of Coagulation Factor Xa Inhibitors

Objective:Through computer virtual design,we investigate the binding mode of coagulation factor Xa active pocket in theory.Based on the theoretical research,we design and synthetize new series compounds TY602.Then we use animal models and in vitro experiment to observe whether TY602would inhibit coagulation factor Xa and what the effect on arterial-venous thrombosis.Methods:Under the condition of computer virtual design,we apply launched and clinical phase coagulation factor Xa inhibitors as the template,by using molecular docking and molecular dynamics,we design and synthesis new series compounds TY602.Screening natural plants database,we made FXa protein receptor and Forsythoside D as a reasonable compound by using molecular docking.In order to investigate the interaction of the ligand and receptor during the dynamic state,we chose500ps molecular dynamics simulation to estimate their binding mode.We apply the model of arteriovenous shut to evaluate whether part of the series compounds TY602has the influence on rat thrombosis.Study the effect of series compounds TY602on prothrombin time (PT),activated partial thromboplastin time (aPTT), thrombin time (TT),fibrinogen content (Fg).And Human clotting factor Xa enzyme-linked immunosorbent assay kit was used to detect inhibitory activity on FXa by series TY602compound in vitro.Results:On theoretical aspect:①We find coagulation factor Xa receptor1FOR,2P16and2UWP have a high G score with template drug and the ligands that we had synthetized,the conformation matching is rather good.The dynamic calculation indicate that receptor2P16and its ligands have a stable binding conformation. Molecular docking screens more than10ligands whose activity is similar to template drug,and it can give the chemical synthesis for further guidance;②Through the molecular dynamics research, we find that RPR remains stable matching state with S1domain and middle part of the receptor depending by hydrogen bond,forms fine spacial matching with S4(Tyr9、Phe174、Trp215) relying on nonpolarity aromatic groups.The interaction style of FTD and receptor is different from RPR,it has weak role with S1domain but has relatively strong action with S4and middle part,as a result,the ligand was repelled from the receptor.It hints that the structure of FXa inhibitor which can combine with S1domain of receptor should be relatively rigid and it can form strong hydrogen bond with receptor.In the S4domain of the receptor, it needs some hydrophobic,aromatic groups and some hydrogen bond in the middle part to help itself fix the ligand.On animal experiment aspect:After oral administration of series compound TY602with the dosage of100mg-kg-1,thrombin was inhibited on arteriovenous shut model.The code of the compound named HYF-12, HYF-8and HYF-6show some inhibition of the role of thrombosis,Inhibition percentage of thrombosis are42.9%,35.0%,29.3%,respectively.Among them,HYF-12has the best effct(p<0.01).After lingual vein administration of series compound TY602with the dosage of30mg-kg-1,PT shows some effects of the extension of clotting time,compared with the blank control group,compounds HYF-4and HYF-6have statistical significance(p<0.05).This result manifested that part of series compound TY602may have the inhibition of thrombosis formation on extrinsic coagulation pathway.The result of ELISA showed that compound named HYF-1, HYF-3,HYF-11,HYF-12,HYF-14,HYF-15,HYF-17and HYF-18have a better inhibition of FXa than launched drug rivaroxaban(p<0.05),HYF-12has the highest rate of enzyme inhibition.Conclusion:Part of series compounds TY602have the inhibitory effect on rat arteriovenous thrombosis,and also have anticoagulant effect.Coagulation factor Xa inhibitors can manage the thrombotic disease, it has extraordinary prospect in clinical application.