Study On The Interaction Between Compound Danshen Dropping Pill And Ginkgo Leaf Tablets Based On Pharmacokinetics And Intestinal Absorption Mechanism

Compound Danshen Dropping Pill is a kind of traditional Chinese medicine dropping pill,which is mainly composed of panax notoginseng,borneol and salvia miltiorrhiza.It is mainly made according to the traditional theory of traditional Chinese medicine and by means of modern medical technology.Ginkgo Leaf Tablets are made from the extract of the dried leaves of Ginkgo(Ginkgo biloba 1.).Clinically used for the treatment of coronary heart disease,angina pectoris,cerebral ischemia and peripheral vascular diseasesClinically in patients with coronary heart disease angina pectoris,cerebral infarction on the basis of conventional treatment given Compound Danshen Dropping Pill and Ginkgo Leaf Tablets combined treatment,the results showed that combination can promote coronary heart disease angina pectoris symptoms,reduce the number of patients with seizure frequency and duration,the degree of ecg ST segment down,cardiac function indexes and improve hemorheology indexes were better than the control group;The improvement effect of nerve function defect and nerve function defect was better than that of control groupTo study the pharmacokinetic and intestinal absorption mechanism of Compound Danshen Dropping Pill combined with Ginkgo Leaf Tablets,so as to understand the interaction between drug absorption and distribution of the two drugs after they enter the body,and provide the corresponding scientific basis for rational drug use and drug administration scheme design in clinical practiceOBJECTIVE:In this paper,the pharmacokinetic rules of Compound Danshen Dropping Pill combined with Ginkgo Leaf Tablets,alone and repeated administration were studied in rats,and the pharmacokinetic parameters were analyzed to understand the changes of pharmacokinetic rules in vivo after combined administration.The rat intestinal perfusion model was used to simulate the absorption of drugs in the human intestinal tract,so as to provide a theoretical basis for the drug administration scheme design and clinical rational use of the combined drugs for the treatment of cardiovascular and cerebrovascular diseasesMETHODS:1.Establish analysis methodology:In vivo high performance liquid chrom-atography(HPLC)of Danshensu,Tanshinone IIA and Quercetin was developed to determine the precision,stability and recovery of RSD2.Pharmacokinetic experimental methods:Compound Danshen Dropping Pill and Ginkgo Leaf Tablets were administered separately and in combination,and rats were given the drugs by single and multiple gavage,in which the time points of administration were consistent.Dosing after 5 min,10 min,15 min,30 min,1 h,2 h,4 h,6 h,8 h,12 h,16 h and 24 h,the plasma of rats were taken,heart,liver,spleen,lung,kidney,and brain tissue,protein precipitation process of plasma and tissue samples,HPLC determination,after processing with DAS3.0 pharmacokinetic software processing,calculation Danshensu,Tanshinone IIA and Quercetin pharmacokinetic parameters and different time points of each group of drug distribution in the data3.Experimental methods of gastrointestinal absorption mechanism:the rats were divided into three groups:Compound Danshen Dropping Pills irrigation group,Compound Danshen Dropping Pills combined with Ginkgo Leaf Tablets irrigation group,Compound Danshen Dropping Pills combined with verapamil irrigation group Abdominal surgery was performed to separate the experimental intestinal segment After washing,both ends of the intestinal segment were inserted into the silicone tube and ligated with wires to connect the constant-current pump.At 0,0.5,1,1.5,2,2.5 and 3 hours,2.0 ml of perfusion fluid was removed from the perfusion bottle respectively,and then 2.0ml of blank phenol red K-R solution was added to keep the volume unchanged.The length and inner diameter of the intestine were measured The intestinal perfusion fluid samples were treated,the concentration was determined by HPLC,the absorption rate constant(Ka)apparent absorption coefficient(Papp)was calculated,and the intestinal absorption mechanism was preliminarily understood.RESULTS:1.Methodological results of HPLC analysis in vivo:The regression equation of the standard curve of Danshen in plasma,heart,liver,spleen,lung,kidney and brain was y=35246x+351.03(R2=0.9998),y=29457x+555.13(R2=0.9998),y=32780x+50.378(R2=0.9991),y=27763x+953.80(R2=0.9988),y=29172x-84.378(R2=0.9993),y=28960x+1488.3(R2=0.9994),y=26957x+535.95(R2=0.9989),precision RSD value?15%,stability RSD valued 15%,recovery RSD value?15%,the results meet the methodological requirements.Tanshinone ?A in plasma and heart,liver,spleen,lung,kidney,and brain tissue of the standard curve regression equation was y=58138x+25.549(R2=0.9985),y=56181x-613.46(R2=0.9982),y=62498x-637.96(R2=0.9981),y=52764x+154.75(R2=0.9984),y=55335x-344.84(R2=0.9988),y=60305x+1111.4(R2=0.9971),y=63786x+933.66(R2=0.9983),the precision RSD value? 15%,the stability RSD valued 15%,the recovery RSD value? 15%,the results meet the methodological requirements;The regression equation of the standard curve of quercetin in plasma,heart,liver,spleen,lung,kidney and brain tissues was y=27047x+267.2(R2=0.9981),y=26342x+399.58(R2=0.9982),y=25542x+684.94(R2=0.9990),y=22674x+426.88(R2=0.9974),y=24672x+838.15(R2=0.9982),y=28758x+1203.2(R2=0.9973),y=24678x+335.05(R2=0.9962),the precision RSD valued 15%,stability RSD valued 15%,recovery RSD value?15%,the results meet the methodological requirements.2.The pharmacokinetic parameters of Danshensu:(1)After a single dose,the plasma peak time(Tmax)was 4.13h;Peak concentration(Cmax)was 0.77?g·mL-1;The area under the curve(AUC)was 5.55?g·mL-1·h;(2)After repeated administration,Tmax was 2.86h;Cmax was 1.18?g·mL-1;AUC was 9.40?g·mL-1·h;(3)After repeated combination,Tmax was 2.15h;Cmax was 1.86?g·mL-1;AUC was 14.99 ?g·mL-1·h;Kinetic parameters of Danshensu(after different administration):the drug concentrations in plasma and tissues of Compound Danshen Dropping Pill combined with ginkgo biloba leaves were all higher than that of separate administeration,and the effect of multiple administration was better than that of single administration.The results of DAS 3.0 pharmacokinetic software showed that danshensin was a two-compartment model in plasma and tissues.The concentration of the drug increased relatively significantly after the combination of brain tissue and liver tissue3.The pharmacokinetic parameters of Tanshinone ?A(1)After a single dose,Tmax was 2.74h;Cmax was 0.22?g·mL-1;AUC was 2.24?g·mL-1·h;(2)After repeated administration,Tmax was 1.33h;Cmax was 0.31?g·mL-1;AUC was 3.19 ?g·mL-1·h;(3)After repeated combination,Tmax was 1.05h;Cmax was 0.52?g·mL-1;AUC was 4.58 ?g·mL-1·h;Kinetic parameters of Tanshinone ? A(after different administration)Tanshinone ? in various tissues are A second chamber model,The concentration of the drug increased significantly after the combination of plasma and brain tissue4.The pharmacokinetic parameters of Quercetin(1)After a single dose,Tmax was 4.53h;Cmax was 0.14?g·mL-1;AUC was 1.25?g·mL-1·h;(2)After repeated administration,Tmax was 3.25h;Cmax was 0.18?g·mL-1;AUC was 1.91 ?g·mL-1·h;(3)After repeated combination,Tmax was 2.3lh;Cmax was 0.23?g·mL-1;AUC was 2.55 ?g·mL"1·h;Quercetin was a two-compartment model in all tissues.The concentration of the drug increased significantly after the combination of heart tissue and lung tissue5.Compound Danshen Dropping Pill combined with Ginkgo Leaf Tablets intestinal perfusion group with Tanshinone ? the absorption rate constant(Ka)and apparent absorption coefficient(Papp)compared with Compound Danshen Dropping Pill intestinal perfusion group had significantly increased,played similar to P-glycoprotein inhibitor verapamil.Ka and Papp of Danshensu decreased in the order of duodenum,jejunum,colon and ileum.Compared with the compound salvia miltiorrhiza group,the absorption parameters of the other two groups were improved The data of duodenal and jejunal segments of the compound salvia miltiorrhiza and Ginkgo Leaf Tablets f group were significantly different from that of the compound salvia miltiorrhiza group(P<0.05).Tanshinone ?A Ka and Papp in accordance with the order of the duodenum,jejunum,ileum and colon decreased,compared with compound salvia miltiorrhiza group,Compound Danshen Dropping Pill combined verapamil group Ka and Papp values increase obviously,and ileum and heartbroken data has significant difference(P<0.05),compared with compound salvia miltiorrhiza group,Compound Danshen Dropping Pill combined Ginkgo Leaf Tablets group in the duodenum and jejunum segment data has significant difference(P<0.05).CONCLUSIONS:1.HPLC analysis in vivo Danshensu,Tanshinone ? A and Quercetin were accurate and reliable,and the results were reliable.2.Ginkgo Leaf Tablets changed the kinetic parameters of the pharmacodynamic components(Danshensu and Tanshinone ? A)of Compound Danshen Dropping Pill in rats,and increased the bioavailability and efficacy.3.Ginkgo Leaf Tablets can promote the absorption of Compound Danshen Dropping Pill in the intestinal tract,and the mechanism may be related to quercetin as a p-glycoprotein inhibitor.INNOVATION:1.The tissue distribution and pharmacokinetic parameters of Compound Danshen Dropping Pill and Ginkgo Leaf Tablets were studied,which provided a basis for further clinical pharmacokinetic research,rational drug use and drug delivery scheme design.2.The absorption of Compound Danshen Dropping Pill combined with Ginkgo Leaf Tablets in the intestinal tract was studied,and it was proved that Ginkgo Leaf Tablets had a promoting effect on the absorption of Compound Danshen Dropping Pill.