Preparation Of Goulou-Xiebai Floating Pellets And Its Antithrombotic Effect

Objective:Preparation of Gualou-Xiebai intragastric floating pellets?abbreviated as pellets?,establishment of a method for measuring in vitro release of pellets,and study of its in vitro release characteristics;High performance liquid chromatography-tandem mass spectrometry?LC-MS/MS?was used to study the pharmacokinetics of pellets in rats,to establish a rat arteriovenous bypass thrombosis model,and to study the antithrombotic effect and mechanism of pellet.Methos:1)Using the floating time of pellets as indicator and the percentage of microcrystalline cellulose?MCC?,octadecanol,hydroxypropyl methylcellulose-K4M?HPMC-K4M?,hydroxypropyl methylcellulose-K15M?HPMC-K15M?in the solid excipients of the prescription as affecting factors on the basis of single factor experiments,Box-Behnken response surface method was used to optimize the prescription.Taking the content of 3,29-dibenzoyl karounitriol in the release medium as the index,the release rate of the pellets in vitro was determined and the release characteristics were explored,and the quality standards of the pellets were established.2)A phenomenex liquid chromatography column?50×4.60 mm,2.6?m?was used,the mobile phase was methanol-0.1% formic acid solution?93:7?,the flow rate was 0.5ml·min^-1,the column temperature was 35 ? and the injection volume was 5 ?L.The multi reactive ions?MRM?in the positive ion mode was detected by using the AJS ESI ion source based on Agilent jet flow technology with the internal standard ofbetulonicacid.The MRM scanning ion pairs of 3,29-dibenzoyl karounitriol and betulonicacid were m/z 689.3?567.2 and m/z 477.2?209.0,respectively.The rats were divided into a pellet group and a Gualou-Xiebai extract group for single administration,and blood was taken from the orbit.The pellet group was at 0,0.5,1,2,3,4,6,8,10,12,24,48 h blood was taken from the orbit.And the Gualou-Xiebai extract group was at 0,5,10,30,60,90,120,180,240,360,480 min blood was taken from the orbit.And the content of 3,29-dibenzoyl karounitriol in rat plasma was measured after administration.3)Rats were divided into normal group?saline,3.0m L/kg?,model group?saline,3.0m L/kg?,Gualou-Xiebai extract group?0.33g/kg?,positive control group?compound Danshen Drpping Pill group,0.09g/kg?,high-,medium-,and low-dose pellet groups?4.0,2.0,0.4g/kg?,which were administered orally for 7 consecutive days,and 8 hours after the last administration,arteriovenous thrombosis was established.by measuring the dry weight of thrombus in each group and measuring the thromboxane B2?TXB₂?and 6-keto-PGF_1? in the serum of each group by enzyme-linked immunoassay?ELISA kit method?to study the antithrombotic effect and mechanism of pellets.Results:1)The optimum prescription of Gualou-Xiebai intragastric floating pellets is as follows: Gualou-Xiebai dry extract 25 g,MCC 6g,octadecanol 11 g,HPMC-K4 M 3g,HPMC-K15 M 11g,70% ethanol solution as wetting agent,40% polyvinylpyrrolidone K30?PVP-K30?solution as binder.The floating time in vitro of the pellets prepared by this prescription was 12 hours,and the cumulative release rate data were fitted well with the first-order release model?r>0.97?.The linear relationship between 3,29-dibenzoyl stilbene triol in the pellets ranged from 0.001 to 0.032 mg/m L?r=0.9999?;precision,stability,repeatability,and sample recovery test RSD of them exceeded 2.0%,and the average recovery was 98.2%.2)The linear relationship of 3,29-dibenzoyl karounitriol was good in theconcentration range of 0.125-0.512 ng/m L?r=0.9925?.The average recovery rate of quality control samples was more than 90%,and the precision within and between days meet the requirements?RSD<15%?.After intracgastric administration of pellets and extracts to rats respectively,the pellets group compared with the extract group,Tmax?8.00±0.00 h?and t1/2z?11.136±2.555 h?of 3,29-dibenzoyl karounitriol in the plasma were significantly prolonged?P<0.01?,Cmax?142.12±14.831 ng/m L?was decreased,and AUC?0-t??1317.853±181.372 ?g/L·h?was increased.3)Compared with the model group,the dry weight of rats in each administration group was reduced,and the results were significantly different?P<0.05?.Compared with the normal group,the TXB₂ content in the model group increased?P<0.05?,the6-keto-PGF_1? content decreased?P<0.05?,and the TXB₂/6-keto-PGF_1? ratio increased,with a significant difference?P<0.01?.Compared with the model group,the TXB₂/6-keto-PGF_1? ratio of each administration group increased?P<0.05?,the TXB₂ content of the compound Danshen Drpping Pill group decreased?P<0.05?,and the6-keto-PGF_1? content increased?P<0.01?.TXB₂ content was reduced in the high and medium dose groups?P<0.05?,and 6-keto-PGF_1? content was increased?P<0.05?;TXB₂ content was reduced in the low dose group?P<0.01?,and 6-keto-PGF_1? content increased?P<0.01?.Compared with the guava white extract group,the TXB₂ content was reduced and the 6-keto-PGF_1? content was increased and TXB₂/6-keto-PGF_1? ratio decreased in each dose group,among them,the high and medium dose groups were statistically significant?P<0.05?.Compared with the normal group,the ratio of TXB₂/6-keto-PGF_1? in the compound Danshen Drpping Pill group and the Gualou-Xiebai extract group increased,with significant differences in results?P<0.05?,and the ratio of TXB₂/6-keto-PGF_1? in each dose group of the pellets to the normal group was closer,and the difference was not statistically significant?P>0.05?.Conclusion:The prescription and preparation process of Guabai Baiwei floating pellets are reasonable.The in vitro release of pellets meets the requirements ofthe Chinese Pharmacopoeia?2015 edition?.Compared with Gualou-Xiebai extract,the pellets have a better sustained-release effect and antithrombotic effect in rats,and the mechanism of antithrombotic effect may be inhibiting platelet aggregation and improving vascular endothelial cell function.