Studies On Therapeutic Basis And Co-pharmacokinetics Of Traditional Chinese Medicinal Naodesheng

Naodesheng, consisted of radix notoginseng, rhizoma chuanxiong, flos carthami, radix puerariae lobatae and fructus crataegi, is one of traditional Chinese medicine recipe for dredging the meridian passage and activating blood circulation to dissipate blood stasis In this dissertation, Naodesheng was systematically studied with Pharmacokinetics (PK) and pharmacodynamics (PD) methods to illuminate therapeutical basis, and to explore the rule of compatibility and the mechanism of effectiveness of the traditional Chinese recipe.By using modern techniques, active sites (active components groups) of crud drugs were isolated and prepared into Naodesheng injections. On the base of quantitative analytical methods for ginsenoside Rg₁, Rb₁, safflor yellow A and puerarin, the HPLC fingerprint chromatograph of Naodesheng injection was developed, in which 35 common peaks were standardized and 13 characteristic peaks were identified. The methods for simultaneous determination of multi-components in Naodesheng injection were developed, including HPLC-UV method for gensenoside Rg₁, Rb₁ safflor yellow A, ferulic acid and puerarin, and HPLC-MS method for gensenoside Rb₁, Rd, Re, Rg₁, Rg₂, Rh₁, notoginsenonide R₁, safflor yellow A, puerarin and daidzein.Simultaneous determination of multi-compounds in rat plasma were developed and were applied to pharmacokinetic research, including HPLC-UV method for safflor yellow A, puerarin, 3'-hydroxyl-puerarin (P3), 3'-methoxyl-puerarin (P4), puerarinapioside (P5), daidzeine (P6), daidzein-7,4'-diglucoside (P2) and HPLC-MS method for safflor yellow A, puerarin, daidzein, ginsenoside Rg₁, Rb₁, Rd, and notoginsenoside R₁. The results showed that by dosage increasing, the pharmacokinetic property of safflor yellow A alter from two-compartment model to one-compartment model while puerarin from approximate two-compartment model to complete two-compartment model; the rate of elimination and the CL of the both decrease when given a high dosage. In the eight of the main flavones, including 3'-hydroxyl-puerarin (P3), 3'-methoxyl-puerarin (P4), puerarinapioside (P5), daidzeine (P6), daidzein-7,4'-diglucoside (P2), unknownâ… (U2), unknownâ…¡(U2) and metabolite (M1), four of them (M1, U1, P4, P5) showed a two-compartment model; P2 showed a significant difference between male and female, the female followed two-compartment model and the male follows one-compartment model; P6 and U2 consistented with one-compartment model; while P3 was consistent with oral first-order absorption process. The plasma concentration of M1 showed a second platform between the 3rd and the 8th hour and P3 showed a peak absorption at the 1st hour. Daidzein, U1, P4 and P5 show similar pharmacodynamic property with puerarin. Ginsenoside Rb₁, Rd, Rg₁ and notoginsenoside R₁ were consistent with the two-compartment model. While ginsenoside Rg₁ and notoginsenoside R₁ had quick distribution and slow elimination rate; ginsenoside Rb₁ and Rd showed slow rate in both distribution and elimination, which may cause aceumulation in vivo. Compared with single chemicals, ginsenoside Rg₁ and notoginsenoside R₁ in Naodesheng injection had quick distribution rate, slow elimination rate and longer elimination half life.After intravenous administration of Naodesheng injection, ginsenoside Rb₁, Rd, Rg₁, Rh₁, notoginsenoside R₁, safflor yellow A, puerarin and daidzein showed a quick distribution into tissue in rat. Safflor yellow A mainly exist in heart, about 3.3 times to serum, 7 times to liver and 2.1 times to kidney. Daidzein was fond in 17 tissues in rat. Although the concentration decreases, in the 1.5th hour daidzein mainly exist in small intestine and stomach with 7.5 times and 5.3 times concentrations compared with that in the 10th minutes respectively. Daidzein was also found in colon with a high concentration. All of this illuminated that daidzein goes through a strong enterohepatic circulation (EHC). Daidzein exists in cerebrum with a similar concentration with that in plasma while the concentration in cerebrum and heart were much higher than given alonly daidzein. Both ginsenoside Rb₁ and Rd go through EHC and ginsenoside Rd, which can be found in lung, heart, uterus, stomach, small intestine at 10th minute after administration and can be also found in most tissues from the 1.5th to the 4th hour, is a metabolite of Rb₁ in intestine. Ginsenoside Rg₁ was found a lot in urinary bladder and kidney but a little in brain 10 minutes after administration, while can only be found in urinary bladder 1.5 hours later and in liver 4 hours later. Ginsenoside Rh₁, widely distributed in rat tissue but has a short residence time, is also a metabolite of Rg₁ in intestine. In the total notoginsenosides, Rb₁and Rd only exist in heart while others exist in both heart and cerebrum.Naodesheng, which has an effect in dredging the meridian passage and activating blood circulation to dissipate blood stasis, is mainly used to heal heart and cerebrum disease caused by blood stasis. This study mainly aims at its effect that dissipates blood stasis. The result of research on rabbit shows that Naodesheng injection has remarkable effects on rabbit's hemorrheology and blood coagulation effect and the effects show a significant dependency on time. The erythroeyte aggregation decreased 30 minutes after administration; the plasma recalcification time (RT) and bleeding time (BT) extend 1.5 hours after injection. 2.5 hours after administration, the clotting time (CT) extended; the hemorheology partly recover 3 hours later while the other effects will stretch to the 6th hour. It was proved that the effect of Naodesheng on rabbit in vivo will lag for about 1 to 1.5 hours and the lag is mainly caused by dynamic process in vivo.In the formula of Naodesheng, safflor yellow (SY) significantly prolonged the recalcifieation time (RT), notoginsenosides (NGs) had assist effect and puerariae flavones (PF) showed negative effect; PF and SY illustrated some effect on hematocrit (HCT) while NGs seems to be negative; effect of NGs on depressing erythroeyte aggregation index (EAI) was noticeable and SY had certain effect on it; PF had a significant effect on heightening erythrocyte deformability, SY had certain effect and NGs shows negative effect on it. The effects of Naodesheng on improving hemorheology index and activity of blood coagulation factor were similar on both rat and rabbit, the effects prolonged the clotting time (CT), RT, and blood viscosity (BV) under high shear pressure seems to be more remarkable for rats than for rabbits. In the recipe, chuanxiong, carthami, pueraria and erataegi had a joint effect with nogoginseng on hemorheology compared with given single NGs to rabbits. Although oral dosage is about 4 times to injection, it showed no better effect because the aceeptor are saturable and the effect shows no dependency on dosage.The combination of serum pharmachemistry, pharmacokinetics and pharmacodynamics is a new way to illuminate the therapeutical basis of traditional Chinese medicine recipe. Correlation analysis was used to analyze the relationship between pharmacodynamics of the main ingredients and pharmacodynamic actions. It was proved that notoginsoside R₁ was the main anticoagulated blood substance, and ginsenoside Rg₁, Rd and Rb₁ had strong effects on anticoagulating blood while daidzein and 3'-hydroxyl-puerarin have some effect on it; daidzein, sail]or yellow A and other puerariae flavones mainly decrease BV while ginsenoside Rb₁ and Rd had certain effect; ginsenoside Rg₁ and notogensenoside R₁ could reduce the "concentration" of blood, and the effect may be connected with depression of the HCT. Metabolite (M1) of Puerariae flavone has a positive effect on reduce of BV, HCT and erythrocyte aggregation, and prolongation of RT, but it showed a negative dependency on the elongation of CT. Ginsenoside Rd, safflor yellow A, daidzein, P2, P3, P4, P5 and puerarin showed negative dependency on reduce of HCT. Gensenoside Rb₁,Rd,Rg₁ and P3 also showed negative dependency on the elongation of CT. All of above show that ingredients in traditional Chinese medicine recipe active cooperatively and restrict each other.Above all, under the direction of Chinese medical science and the practice, this study had conjoined traditional Chinese pharmacology, analytical chemistry, pharmacology and serum pharmaceutical chemistry to develop the quality control method, approach the rule of compatibility, find out the therapeutical basis and illuminate the maehanism of Naodesheng injection. It is a meaningful approach for the way of researching traditional Chinese medicine recipe.