Study On The Antithrombotic Effect And Its Mechanism Of Compound Trichosanthis-aspirin Pellet Capsules

Objective:The purpose of this study was to prepare compound Trichosanthis-aspirin pellet capsules,determine the optimal preparation process,establish the determination methods for the content and release of compound Trichosanthis-aspirin pellet capsules,and explore the antithrombotic effect and mechanism of them.Method:1)To prepare compound Trichosanthis-aspirin pellet capsules.The optimal formulation of aspirin pellets and preparation process were determined by single variable method and orthogonal test.Then the enteric coated layer was prepared by fluidized bed.Using blank pill core prescription method by fluidized bed coating technology preparation of Trichosanthis contain pill core and package film layer.The prescription and process were optimized by orthogonal experiment,and the pill core was prepared by extrusion apheronization.2)HPLC fingerprint of compound Trichosanthis-aspirin pellet capsules was established by HPLC,and the contents of cytosine,guanine,uracil,hypoxanthine,adenosine,guanosine and aspirin were determined.The method was investigated by linearity and range,precision,stability,repeatability and recovery.HPLC fingerprint similarity of trichosanthes kirilowii and aspirin content were used to evaluate the release of compound Trichosanthis-aspirin pellet capsules.3)The rats were divided into blank group,model group,high,medium and low dose group of compound Trichosanthis-aspirin pellet capsules,high,medium and low dose group of trichosanthis pellet,aspirin enteric pellet group,and positive control group was clopidogrel sulfate.These medicine was given to the stomach for seven days,once a day.The rat arteriovenous bypass model was established 1 h after administration on day 8.Through the determination of dry weight of thrombus,detection in plasma thromboxane B2(TXB2),cyclic adenosine monophosphate(cAMP),6-ketone-alpha prostaglandin F1?(6-keto prostaglandin F1?,6-keto-PGF1?)content and prothrombin time(PT),thrombin time(TT),activated partial thrombin time(APTT)and fibrinogen(FIB)coagulation four indicators to evaluate the antithrombotic effect and mechanism of compound Trichosanthis-aspirin pellet capsules.Results:1)The prescription excipient for aspirin containing pill core is microcrystalline cellulose and the disintegrating agent is sodium carboxymethyl starch.The mass ratio is aspirin:MCC:CMS-Na=40:60:3.The 2% hydroxypropyl methylcellulose(HPMC)solution was used as adhesive,with a frequency of 50 Hz and a time of 5 min.The prescription process has good repeatability.2)The similarity of HPLC fingerprint of trichosanthis kirilowii was 0.998.The contents of cytosine,guanine,uracil,hypoxanthine,adenosine and guanosine were 0.11%,0.40%,0.036%,0.015%,0.026% and 0.045%.Aspirin content was 2.1%.The results of methodological investigation showed that the RSD values of precision,stability,repeatability and recovery were all in line with the requirements.Trichosanthis pellet can be completely released in simulated gastric acid environment for 5 min.The cumulative release rate of aspirin enteric-coated pellets was greater than 75% within 45 min in simulated intestinal fluid environment.It complies with the regulations and requirements of Chinese pharmacopoeia(2015 edition).3)Compared with the blank group,TXB2 level was significantly increased in the model group(P<0.01),and 6-keto-PGF1? content was significantly decreased in the model group(P<0.01).Compared with the model group,the TXB2 content(P<0.01),6-keto-PGF1? content(P<0.01),cAMP content(P<0.01)and TXB2/6-keto-PGF1? content(P<0.01)were decreased by trichosanthis.Trichosanthis-aspirin pellet capsules,trichosanthis pellet and aspirin enteric-coated pellets.Conclusion: The compound Trichosanthis-aspirin pellet capsules were reasonable and the preparation process was reproducible.The release degree and acid resistance are in line with the requirements of Chinese pharmacopoeia(2015 edition).The quality standard of compound Trichosanthis-aspirin pellet capsules established can be used for the quality evaluation of this preparation.The compound Trichosanthis-aspirin pellet capsules have the effect of antithrombosis,inhibiting platelet aggregation,reducing vascular endothelial injury and dilating blood vessels.