Design,Synthesis And Evaluation Of Anti-platelet Aggregation Activity Of The Structure Analogues Of Picotamide

Atherothrombosis is a major cause of cardiovascular disease,and platelets play an important role in arterial thrombosis.Anti-platelet aggregation drugs can inhibit the formation of thrombosis that occupy a key position in antithrombotic drugs.TXA₂ inhibitors have been the hotspot of antiplatelet drugs.Since first-line therapies of anti-platelet drugs such as aspirin and clopidogrel still have some limitations.Therefore,to explore some novel efficient antiplatelet drugs with low side effects and ease to use has a very high social and economic prospects.Picotamide is a broad-spectrum antiplatelet drug with dual effects including inhibit the synthesis of TXA₂ and TXA₂/PGH₂ receptor antagonism.PN01?IC50=1.7?mol/L?and PSN01?IC50=0.46?mol/L?are two structural analogues of picotamide.PN01 has an amide structure and PSN01 has a sulfonamide structure.In this paper,PN01 was used as the lead compound to introduce acylhydrazone fragments into the two 1,3-position side chains of PN01 for the first time and nineteen 4-methoxy-1,3-dibenzenedihydrazone compounds PNN01-PNN19 were synthesized.PSN01 used as the lead compound,5 or 6 methyl separately or in the same time were introduced into the parent ring,designed and synthesized three 5-methyl-4-methoxy-1,3-dibenzene sulfonamide compounds PN734-PN736,seven6-methyl-4-methoxy-1,3-dibenzenesulfonamide compounds PN739,PN749-PN752 and PN756-PN758 and four 5,6--dimethyl-4-methoxy-1,3-dibenzene sulfonamide compounds PN738 and PN753-PN755.All the 34 compounds have been confirmed by ¹H NMR,¹³C NMR,IR And ESI-MS spectroscopy.Fries rearrangement method which was designed by our research group was used to synthesis the key intermediate 4-methoxy-1,3-benzenedicarboxylic acid.Compared with other methods,the cost of raw materials in each step is lower and avoiding the use of large doses of strong acids and less damaging to the environment and equipment,the total yield can reach to80%.Born method was used to test the in vitro anti-platelet aggregation activity of the target compounds induced by ADP and AA.The experimental results showed that 29 compounds are higher than Picotamide in inhibition activity and three compounds comparable.When induced by AA,all of the 19 acylhydrazone compounds exhibited better anti-platelet aggregation activity in vitro than the positive control drug and seven of them?PNN01,PNN03,PNN05,PNN07,PNN10,PNN12,PNN14?is higher than aspirin.Two of the fifteen sulfonamides showed higher inhibitory rates than Picotamide,another two of which were comparable.When ADP was used as an inducer,8 compounds with sulfonamide structure were higher than Picotamide,one of which was comparable to the control drug,the inhibition rate of PN755was higher than that of aspirin.The experimental results showed that two kinds of compounds with different structures may choose to act on different inducer pathways that make the research direction and results have the value for further study.The eighteen active compounds selected was carried out in vitro cytotoxicity test in L-929cell.Compound PNN03 with novel diacyl hydrazone structure showed lower toxicity comparable to the positive drug picotamide.The other target compounds showed a relatively high cell viability at the lower concentration?10?mol/L?,The cell viability was relatively lower at higher concentration?100?mol/L?and higher than that of Picotamide.Therefore,the compound PNN07 with high activity,low toxicity,further study is worthy to be done.In addition,the pharmacological test results were analyzed and summarized,and the structure-activity relationship of the target compounds was preliminarily estimated.