| The vascular embolic disease is one of the diseases with higher morbidity and mortality around the world.The arterial thrombosis caused by platelet activation and aggregation is an important pathogeny of this kind of disease.So,platelets function is essential for the occurrence of thrombotic diseases.At present,the combination of clopidogrel and aspirin is mainly used in clinical practice to the prevention and treatment of acute coronary syndrome,myocardial infarction and stroke.However,with the increase of clinical application,the disadvantages of this medication mode have gradually been exposed,such as the low bioavailability of clopidogrel,the existence of"clopidogrel resistance"and so on.At the beginning of the 21st century,a new generation of drugs,Prasugrel,has strong antiplatelet aggregation activity,but it is accompanied by a greater risk of bleeding.Therefore,the development of new antiplatelet drugs with strong efficacy and small side effects has become the first task of current medical researchers.Basing on the antiplatelet effect and current status of thienopyridine P2Y12 receptor antagonists,referencing to the drug structures of clopidogrel and prasugrel,and utilizing selective deuteration technology and group introduction technology,we got a variety of new thienopyridine drugs.The results of previous pharmacokinetic studies have shown that deuterated clopidogrel derivative can produce more active product,avoid CYP2C19 metabolic pathway and overcome clopidogrel resistance,which indicates the significant pharmacokinetic advantages of this drug,this article will conduct a deeper study on its pharmacodynamic activity.Purpose:This article will study the antiplatelet activity and safety of deuterated clopidogrel derivative on the basis of pharmacokinetic test results,in order to solve the problems of low bioavailability and high bleeding risk of traditional antiplatelet drugs,and to provide experimental basis for developing it into a new type of antiplatelet drug for clinical application.Methods:Evaluation of antiplatelet activity:1.Wistar rats were randomly divided into 5 groups,including vehicle group?0.5%CMC-Na?,low dose group of deuterated clopidogrel derivative?0.5mg/kg?,medium dose group?1.0mg/kg?,high dose group?2.0mg/kg?,clopidogrel group?10.0mg/kg?,and there are 8 rats in each group.Optical turbidimetry was used to determine the platelet aggregation rate and maximum aggregation rate within 1,3,and5 minutes after adding ADP,to evaluate the antiplatelet aggregation activity of this drug.2.Male and female KM mice were randomly divided into 5 groups,including vehicle group?0.5%CMC-Na?,low dose group of deuterated clopidogrel derivative?0.75mg/kg?,Medium dose group?1.50mg/kg?,high dose group?3.00mg/kg?,clopidogrel group?15.00mg/kg?,there are 10 mice in each group.?1?The anticoagulant activity of the deuterated clopidogrel derivative was measured by the capillary glass tube method;?2?The bleeding time was measured by the tail-cut method to evaluate the bleeding risk of the drug;?3?Carrageenan was injected intraperitoneally in mice to prepare an in vivo thrombus model and evaluate the inhibitory effect of the drug on thrombosis;?4?The content of 5-HT,P-selectin and cAMP in mouse serum were determined by enzyme-linked immunosorbent assay to evaluate the effect of the drug on the effect of platelet release function,and preliminarily explore the mechanism of antiplatelet activity of this drug.Preliminary safety evaluation:Male and female KM mice were randomly divided into 3 groups,including blank group,vehicle group?0.5%CMC-Na?,and deuterated clopidogrel derivative group?300mg/kg?,and there are 10 mice in each group.The serum biochemical indexes of LDH,ALB,ALT,AST,BUN,CRE,TG and T-CHO were measured by acute systemic toxicity test.HE staining was used to observe the pathological sections of the heart,liver,spleen,lung,kidney and stomach of mice,and to preliminarily evaluate the safety of this drug.Results:?1?Compared with the vehicle group,the maximum platelet aggregation rate of the medium dose and high dose deuterated clopidogrel derivative were significantly reduced?P?0.001?;compared with the clopidogrel group,the maximum aggregation rate of the drug's high dose group was not significantly different.At 1 min,3 min,and5 min of the aggregation experiments,the platelet aggregation rate in each dose group of deuterated clopidogrel derivative showed a dose-dependent decrease trend.?2?Compared with the vehicle group,the clotting time in the low,medium and high dose groups of deuterated clopidogrel derivative were significantly prolonged?P?0.05,P?0.01,P?0.001?,and the clotting time gradually increases with the increase of dosage;compared with the clopidogrel group,there was no significant difference in clotting time among all dose groups of deuterated clopidogrel derivative.?3?Compared with the vehicle group,the three dose groups of deuterated clopidogrel derivative significantly prolonged bleeding time?P?0.001?;compared with the clopidogrel group,the bleeding time of the deuterated clopidogrel derivative low dose and medium dose group was significantly shortened?P?0.01,P?0.05?.?4?The thrombus model in mice was prepared for 24h,compared with the vehicle group,the tail thrombus length of the deuterated clopidogrel derivative medium and high dose groups was significantly shortened?P?0.05?;compared with the clopidogrel group,there was no significant difference about the thrombus length in each dose group of deuterated clopidogrel derivative;there was no significant difference in the thrombus length of 48h's model compared with that of 24h.?5?Compared with the vehicle group,the 5-HT content in all dose groups of deuterated clopidogrel derivative was significantly reduced?P?0.05,P?0.05,P?0.01?,the P-selectin content of its medium and high dose groups was significantly reduced?P?0.05,P?0.01?,and the cAMP content of its medium and high dose groups was greatly increased?P?0.05?;compared with the clopidogrel group,there was no significant difference in 5-HT content and cAMP content between all dose groups of deuterated clopidogrel derivative,and so did the P-selectin content of this drug's medium and high dose groups.?6?The results of the acute systemic toxicity test showed that there was no significant difference in LDH,ALB,ALT,AST,BUN,CRE,TG,T-CHO levels in the serum of the blank group,the vehicle group and the deuterated clopidogrel derivative group;the result of HE staining showed that there were no significant pathological changes in the heart,liver,spleen,lung,kidney and stomach tissues of all animals.Conclusion:Deuterated clopidogrel derivative showed strong antiplatelet activity and inhibition of thrombus formation,there is no tendency to increase the risk of bleeding while exerting the anticoagulant effect,the mechanism may be related to the decrease of 5-HT and P-selectin release and the increase of cAMP content in serum,in addition,the drug has no obvious effect on the main organs such as heart,liver,kidney,etc.therefore,it has a good application prospect. |
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