The Effect Of Zhengtian Pill On The Activity Of Drug Metabolizing Enzymes

ObjectiveBy observing the effect of Zhengtian pill hypnotic effect of pentobarbital sodium andsodium pentobarbital, understanding Zhengtian Pill on liver microsomal cytochrome P450activity had no inhibitory effect. Further incubation in vitro reaction, P4503A4of Zhengtian Pill on rat liver microsomal cytochrome (CYP3A4) activity, and calculate the IC50,based in vitro experiment, effect of Zhengtian Pill on carbamazepine in drug pharma-cokinetics parameters in vivo in rats. Based on the above results, further determination ofcontent of Zhengtian pill of total coumarins and part of main coumarins. To provide theor-etical basis and experimental data for clinical drug interactions, adverse reaction of treatm-ent, reduce side effects and reduce the treatment costs, to ensure the rational use of drugs inthe clinic.Methods1、60mice were randomly divided into control group (2%Twain-80,20mL·kg-1), pos-itive chloramphenicol group (25mg·kg-1) and Zhengtian pill low, high dose (3,6g·kg-1)group,15rats in each group, continuous isometric by gavage for3days, the last drug1hafter intraperitoneal injection to mice, and then by barbital sodium pentoba-rbital sodium40mg·kg-1or180mg·kg-1, the mice sleep latency and sleep time.2、the prep-aration of rat liver microsomes, the establishment of in vitro incubationreaction system, with midazolam as probe drug, determination of midazolam metabolite1’-hydroxy midazolam (1’-OH-MDZ) concentration. Incubation were divided into controlgroup, six concentration test group and positive chloramphenicol group eight group,control group as a probe drug reaction, the test group was six days effective componentconcentrations, respectively0.625,1.25,2.5,5,10and20mg·L-1. Probe drugs with the pipewith CYP3A4Midazolam (MDZ) after incubation of20min reaction, the ice bath coolingend reaction. A RP-HPLC method for the determination of in vitro incubation in reaction system probe drug metabolite1’-hydroxy midazolam (1’-OH-MDZ) final concent-ration.According to the different concentration of inhibition rate, calculated IC50, determi-ne theinhibitory effect of Zhengtian pill effective composition on CYP3A4activity of rat livermicrosome strength.3、take the health of12Wistar rats, were randomly divided into2groups. Give medic-ine group were given4g·kg-1Zhengtian pill suspension gavage,1times a day, continuous5days; the control group was given equal volume of physiological saline. The last admini-stration of1H, the rats were given intragastrically to carbamazepine50mg·kg-1carbamaz-epine, after intragastric administration of0.25,0.5,1,1.5,2,3,5,7,10,15and24h from t-ail vein blood0.5ml, centrifugal plasma, using high performance liquid chromatographymethod for the determination of carbamazepine groups. The drug blood concentration,pharmacokinetics parameters and calculating carbamazepine in rats.4、Selection of5batches of Zhengtian pill, extraction of total coumarins Zhengtian pi-ll in, with imperatorin as control sample, using ultraviolet spectrophotometry, the detectionwavelen-gth was300nm, the determination of total content of coumarin in days.Results1、chloramphenicol group and the low, high doses of Zhengtian pill group thepentobarbital sodium on mice sleep time extension222.2%,123.1%,42.6%respectively,the chloramphenicol group and small dose of Zhengtian pill group and solvent controlgroup, mice sleep time was significantly prolonged (P<0.01). Zhengtian pills high dosegroup hypnotic time less than Zhengtianwan pill hypnotic time low dose group, there wasno significant difference of high dose group and the control group (P>0.05). High doses ofZhengtian pill could significantly prolong the latency (P<0.01) sodium pentobarbitalinduced hypnosis, hypnosis and shorten the time (P<0.01). Chloramphenicol group andlow dose group of Zhengtian pill hypnotic effect on the latency and time, and the controlgroup showed no significant difference. The final concentration of metabolites of1’-OH-MDZ probe drug Mi.2、blank control group of midazolam was2.62μmol·L-1. The positive control drug chl-oramphenicol six concentrations were strongly inhibited the1’-OH-MDZ formation rate,and there was a dose dependent inhibition (P<0.01). Six the concentration of effective co-mponents of Zhengtian Pill on rat liver microsomal CYP3A4showed different degrees ofinhibition, and also in a dose-dependent manner (P<0.01). IC50days effective componentson the activity of CYP3A4was8.99mg·L-1.3、two groups of carbamazepine mainly drug Pharmacokinetics parameters: medicat- ion group and peak time of Tmax (3.756±1.460), peak concentrations of Cmax (13.633±4.20),Area under concentration time curve of AUC (183.918±59.640), elimination half-life of T1/2ke (7.362±2.521), elimination rate constant Ke (0.833±0.850), the total removalrate in CL/F (0.3435±0.144), the apparent volume of distribution of V/F (3.774±2.095); gr-oup Tmax (2.057±1.795), Cmax (10.250±7.994) AUC,(162.788±18.247) T1/2ke,(5.710±2.677) Ke,(9.352±14.906) CL/F,(0.6735±0.584),(4.885±4.736) V/F. With Zhengtian pi-ll treatment group rats, compared with the control group of rats, the elimination rate const-ant (Ke carbamazepine) and total body clearance (CL) decreased significantly, the elimin-ation half-life (T1/2Ke) extension, the area under the curve (AUC) increased, the peak con-centration (Cmax) were significantly increased(P<0.05).4、Different batches of Zhengtian pill of total coumarin content were1.262±0.033,1.104±0.032,1.288±0.024,1.464±0.036,1.476±0.056.Conclusion1、Zhengtian pill can inhibit the liver microsomal cytochrome P450activity, also haveexcitatory effects on the central nervous system of mice.2、Zhengtian pill of alcohol extracted components on CYP3A4activity of rat livermicrosomes showed inhibitory effect, IC508.99mg L-1.3、Zhengtian pill can inhibit carbamazepine in rats in vivo metabolism, eliminate thespeed slowed down, blood drug concentration increased significantly.4、Zhengtian pill of total coumarin content of1.32%.