Pharmacodynamic And Pharmacokinetic Study On DXF And Interaction Of Its Components

Our lab had designed a series of new Chinese medical complex prescription, which was composed by water soluble components from danshen, liposoluble conmponents from danshen and ferulic acid from Chuanxiong. It was named by DanXiongFang1, 2, 3, 4, 5, 6 (DXF1, 2, 3, 4, 5, 6). In order to convenient for quality control, we had selected danshensu as the target chemical of water-soluble components, total tanshinone as the pharmacodynamic target and cryptotanshinone as the pharmacokinetic target of the liposoluble conmponents from danshen. We had carried out a preliminary pharmacodynamic screen of the prescription with C57BL/6J hyperlipidemia model induced by high-lipid diet, and selected DXF1and DXF4, then a series of pharmacodynamic and pharmacokinetic study was carried out.1.Effect of Danxiongfang on reducing serum lipids level in rat model with hyperlipidemia induced by high-lipid diet: The rats were fed with high-fat diet for 4 weeks to establish the hyperlipidemic model, then lovastatin(7mg/kg), cholestyramine(1300mg/kg), DXF1 330, 66, and 13mg/kg were given to the rats orally for 4 weeks. The serum TG, TC, HDL, LDL, ALT, albumin were detected at the end of 4 weeks. The results showed that the rats developed hyperlipidemia with high serum lipid levels (TC: 2.08±0.24 vs 16.07±4.13mmol/L; LDL: 0.42±0.10 vs 7.57±2.33 mmol/L; TG: 0.74±0.10 vs 1.18±0.71mmol/L), serum ALT and albumin concentration also increased significantly. DXF1 330mg/kg could decrease the serum TC to 6.78±3.82mmol/L, better than lovastatin (13.04±4.60mmol/L) and similar to cholestyramine (8.43±4.64mmol/L). There showed similar effect on LDL. It is noteworthy that DXF1 13mg/kg showed the strongest inhibitory effect on serum TG levels. At the same time they all could decrease the ALT level and increase serum albumin level. A conclusion could be drawn that DXF posses powerful lipid regulating and liver protective effect.2.Effect of DXF and its active components on Experimental liver injury models induced by CCl4 in mice: Mice are injected with CCl4 to establish liver injured model. ALT, AST, serum albumin, globulin in serum and SOD, MDA in liver are measured to confirm the ability of protecting liver of DXF4 (247.1, 49.42, 9.88, 1.97mg/kg), water-soluble components 20.7mg/kg, liposoluble components 12.6mg/kg,ferulic acid 16.12mg/kg. Bifendate pills was selected as positive drug, its dose was 25.6mg/kg. The results showed DXF4 and all components could inhibit obviously the abnormal increase of ALT, AST in serum and MDA in liver, enhance SOD activity in liver, total protein, albumin, globulin in serum. DXF4 247.1 and 1.97mg/kg exhibited the strongest effect. The results suggested that DXF4 not only posses the potentialities of protecting injured liver induced by CCl4, but also optimize the action.3.Anti-inflammatory effect of DXF: Set up acute animal models, such as egg white induced paw oedema in rat, sodium carboxymety-lcellulose (CMC-Na) induced air pouch leukocyte migration in rat, and a chronic inflammatory reaction model placing cottons into abdominal cavity of rats to induce granuloma to test anti-inflammatory activity of DXF1 (330, 66, 13mg/kg), prednisone acetate 1mg/kg was positive drug. The results showed that DXF1 330, 66mg/kg could inhibit paw oedema, leukocyte migration and the proliferation of granuloma, they took action faster than prednisone acetate. DXF1 66mg/kg had stronger effect than 330mg/kg, but 13mg/kg hadn't exhibit any effect on three inflammatory models. The evidences suggested that DXF1 has definit anti-inflammatory effect.4.Protective effect on brain in mice under normobaric hypoxia station: 60 mice were divided randomly and equally into 6 groups, they were control group, model group, propranolol group(12mg/kg) and DXF4 groups(160, 60, 30mg/kg), All reagents were given intra-gastrically into each mouse once a day for 7 days, at last day of the study, the mouse was put into a sealed bottle to observe the survival time of the mouse, the levels of MDA, SOD, Lac and NOS in the brain were detected with test kit. The results showed that DXF4 160mg/kg could prolong the survival time of the mouse significantly; but all groups could obviously inhibit the abnormal increase of MDA and Lac in brain, and reduce NOS activity in brain, and enhance SOD activity in brain. These results showed that DXF4 posses the protective effect against brain injury under normobaric hypoxia station.5.Pharmacokinetic characteristics and interaction of main active components in DXF: A new RP-HPLC/DAD method was established to simultaneously determine the biomarker content of danshensu, ferulic acid and cryptotanshinone in DXF. Then the pharmacokinetics characteristics of the main active components danshensu, ferulic acid, cryptotanshinone in rabbits were investigated after i.v. administration any components alone or DXF by the method of HPLC. The pharmacokinetical results indicated that the plasma drug concentration-time curves of danshensu, ferulic acid and cryptotanshinone in rabbits were all best fitted to two-compartment open models after i.v. component alone or DXF. The results showed that danshensu and ferulic acid were distributed and eliminated comparatively fast in rabbits, their t1/2αwere 2.84±2.00min and 1.47±1.30min, and t1/2βwere 20.95±10.18, 19.28±10.21min. The distribution process of cryptotanshinone was also fast, t1/2αwas 2.13±0.72min, but its elimination process was 3 times slower than danshensu and ferulic acid, its t1/2βwas 69.69±27.22min. Administration of compound prescription had no influences on distribution and elimination process within body of danshensu and ferulic acid, but it could prolong the t1/2βof cryptotanshinone to 85.01±42.46min. The AUC of ferulic acid and cryptotanshinone were obviously improved from 94.33±27.65μg·min/ml and 80.49±19.21μg·min/ml to 122.05±48.92μg·min/ml and 129.34±44.74μg·min/ml. As a conclusion it can be said that DXF could optimize the pharmacokinetic process in vivo of target chemicals from those components.6.Absorption characteristics and interaction of DXF and its main active components: In order to study the absorption characteristics of DXF4 and its components, we established a model of in situ perfused rat intestine preparation and a HPLC-MS method to determine the concentration of danshensu, ferulic acid and cryptotanshinone. The dose of DXF4 was 164mg/kg (danshensu 69 mg/kg, ferulic acid 53.7 mg/kg, total tanshinone 42.5 mg/kg), the doses of components in DXF4 were similar to that in DXF4. 200μl samples were collected at 15, 45 and 90 minute, the concentration of target chemicals were detected by HPLC-MS. The results showed that DXF4 have no effect on absorptive permeability of ferulic acid and cryptotanshinone, but can increase the absorptive permeability of danshensu.