Preclinical Pharmacokinetics Of Pseudo - Berberine Pseudo - Berberine Pseudo - Berberine Pseudorabine Pentaphyllum And

In the past few years, compounds of THPBs have drawn global attention due to its wide range of sedation、analgesic effect and excellent potential to be a new drug, but the pharmacological、pharmacokinetics study of this kind of compounds haven’t been explored enough, only l-Tetrahydropalmatine(l-THP) got detail research and finally been a clinical medicine. So, the pharmacokinetics of THPBs and study involving its absorption, distribution, metabolism, excretion and the law of metabolic are very important to the development of these compounds.In this paper, we study on l-corydalmine(l-CDL) and l-Tetrahydrobeberruine(l-THR), An LC-MS/MS method was developed for the quantification of l-corydalmine(l-CDL) and l-Tetrahydrobeberruine(l-THR) to study the pharmacokinetic characteristics of SD rats, we also make preliminary study of l-corydalmine(l-CDL) in sedation、analgesic effect, as well as the comprehensive research of l-corydalmine(l-CDL) and its metabolic product l-Stepharanine(l-SPD) in excretion and distribution in rats. These studies aimed to clarify the pharmacokinetic characteristics of these two kinds of THPBs and its metabolic properities.1. Study the sedative and hypnotic effects of l-corydalmine(l-CDL)Objective: To study the sedative and hypnotic effects of l-corydalmine(l-CDL), preparing for next pharmacokinetic study.Methods: The effect of l-corydalmine(l-CDL) on the locomotion behavior of mice: 40 mice were intraperitoneal injection of three different dose of l-corydalmine(l-CDL) 1、5、10 mg?kg-1, and immediately put mice into the YLS-1 multi-function locomotive behavior recorder after dosing; Recording the number of activities of mice within 90 minutes after dosing and examining the effect of locomotive behavior on mice under different doses. The effect of l-corydalmine(l-CDL) on sedative hypnotic effect of Pentobarbital sodium: 40 mice were intraperitoneal injection of three different dose of l-corydalmine(l-CDL) 1、5、10 mg?kg-1 and solvent as comparison, then giving every mice 40 mg?kg-1of Pentobarbital sodium after 30 minutes, recording the sleeping time and waking up time in each group. The synergy of l-corydalmine(l-CDL) and Pentobarbital sodium under subthreshold hypnotic dose: 40 mice were randomly divided into 4 group, intraperitoneal injection of three different dose of l-corydalmine(l-CDL) 1、5、10 mg?kg-1 and solvent as comparison, then giving every mice 30 mg?kg-1of Pentobarbital sodium after 30 minutes, recorded the number of mice in sleeping.Results: The effect of l-corydalmine(l-CDL) on the locomotion behavior of mice: The activity of mice were slightly increased after giving l-corydalmine(l-CDL) 1 mg?kg-1, but the activity of mice with giving 10 mg?kg-1 were significantly decreased, and the inhibitory effect lasted for 90 minutes. The effect of l-corydalmine(l-CDL) on sedative hypnotic effect of Pentobarbital sodium: The l-corydalmine(l-CDL) could extend the sleeping time of mice with Pentobarbital sodium under subthreshold hypnotic dose, and it could also obviously shorten the incubation sleeping time of mice under the dose of 5 mg?kg-1. The synergy of l-corydalmine(l-CDL) and Pentobarbital sodium under subthreshold hypnotic dose: 1、5、10 mg?kg-1of l-corydalmine(l-CDL) could increase the number of mice in sedative、hypnotic induced by Pentobarbital sodium under subthreshold hypnotic dose.Conclusion: l-corydalmine(l-CDL) is definitely has sedative、hypnotic effect, but its pharmacological mechanism still needs further research.2. The pharmacokinetics of l-corydalmine(l-CDL) in ratsObjective: To develop a LC-MS/MS method for the quantification of l-corydalmine(l-CDL) in rat plasma, and study its pharmacokinetics and bioavailability in rats.Methods: 10 rats were divided into 2 groups, one group was intravenous injected with l-corydalmine(l-CDL) 3 mg?kg-1, after administration 0、2、5、15、30 min and 1、2、4、8、12、24 h, retinal venous plexus blood, blood sample analysis preprocessing backward then set into LC-MS/MS measure its concentration; another group was given l-corydalmine(l-CDL) 10 mg?kg-1of oral administration, after administration 0、5、15、30 min and 1、2、4、8、12、24 h,retinal venous plexus blood, blood sample analysis preprocessing backward then set into LC-MS/MS measure its concentration.Results: The LC-MS/MS method we developed was simple、rapid、high sensitivity、good precision with little matrix effect. The calibration curve was liner across the concentration range of 1~5000 ng?m L-1 for l-Corydalmine(l-CDL) with a lower LOQ of 1ng?m L-1. The within-run and between-run precision(RSD%) was in the range 0.1%~10%. l-Corydalmine(l-CDL)reached the peak concentration at 0.5h after dosing. The main pharmacokinetic parameters were as follows : ig : t1/2(7.04±3.93)h, Cmax(557.78±330.14)ng?m L-1,AUC0~24(2408.01±630.49)ng?m L-1?h, AUC0~∞(2646.76±646.10)ng?m L-1?h;iv:t1/2(4.14±1.66)h,Cmax(1926.13±176.86)ng?m L-1,AUC0~24(1390.34±253.93.Conclusion: l-Corydalmine(l-CDL)has good internal absorption、 long elimination time and high bioavailability.3. Excretion Study on l-Corydalmine(l-CDL)and its metabolite l-Stepharanine(l-SPD) in rats Urine、 Feces and BileObjective: To establish a LC-MS/MS method for determination of l-Corydalmine(l-CDL)and l-Stepharanine(l-SPD) in urine、feces and bile, and to explore the rule of excretion as well as the main pathways of excretion.Methods: Excretion Study on l-Corydalmine(l-CDL) and its metabolite l-Stepharanine(l-SPD) in rats urine and feces: After 6 rats were orally administration of l-Corydalmine(l-CDL) at a single dose 10 mg?kg-1, we collected the samples from urine and feces at different times. Recorded the urine volume and the weight of dried feces. Using LC-MS/MS method to determine the concentration of l-Corydalmine(l-CDL)and l-Stepharanine(l-SPD), and then calculated the accumulation and recovery in urine and feces. Excretion Study on l-Corydalmine(l-CDL)and its metabolite l-Stepharanine(l-SPD) in rats bile: 4 rats were orally administration of l-Corydalmine(l-CDL) at a single dose 10 mg?kg-1 after biliary drainage operation, then collected the bile at the set point time, recorded the bile volume. Using LC-MS/MS method to determine the concentration of l-Corydalmine(l-CDL)and l-Stepharanine(l-SPD), and then calculated the accumulation and recovery in bile.Results: The established method of l-Corydalmine(l-CDL)and l-Stepharanine(l-SPD) in rats urine、feces and bile with good specificity、little matrix effect and high sensitivity. The recovery of l-Corydalmine(l-CDL)in rats urine was only 0.52%, which of l-Stepharanine(l-SPD) was 7.5%; the recovery of l-Corydalmine(l-CDL)in rats feces was 10.5%, while the recovery of l-Stepharanine(l-SPD) was 4.66%; the recovery of l-Corydalmine(l-CDL)in bile was 12.8%, However, the recovery of l-Stepharanine(l-SPD) in bile was 30.2%.Conclusion: The recovery of l-Corydalmine(l-CDL)in urine was very little, while much higher in feces and bile; The recovery of l-Stepharanine(l-SPD) was higher in bile than in urine and feces. Suggested that l-Corydalmine(l-CDL) exist comprehensive metabolism in rat, the reason why l-Corydalmine(l-CDL) was less in urine was that there might be first pass effect in liver.4. Tissue distribution study on l-Corydalmine(l-CDL) in ratsObjectives: To established the LC-MS/MS method for determination of l-Corydalmine(l-CDL) and l-Stepharanine(l-SPD) in rat tissues and investigate the distribution rules.Methods: 30 rats were randomly divided into 5 groups with each group of 6. Only one group given the physiological saline as blank comparison, then collected the tissue samples. And also we collected the tissue samples at different times after the rest 24 rats were orally administration of a single dose 10 mg?kg-1 of l-Corydalmine(l-CDL). Recorded the weight of each tissue sample and determined the concentration of l-Corydalmine(l-CDL) and l-Stepharanine(l-SPD) in each tissue samples by LC-MS/MS after samples were homogenated.Results: l-Corydalmine(l-CDL) was comprehensively distributed in heart、liver、spleen、lung、kidney、brain、intestine and muscle, but the concentration of which was higher in liver、spleen and lung at all time points. l-Stepharanine(l-SPD) was also comprehensively distributed in all the tissue except brain, the concentration of l-Stepharanine(l-SPD) was much higher in intestine and kidney than other tissue samples.Conclusion: l-Corydalmine(l-CDL) and l-Stepharanine(l-SPD) were widely distributed in various tissues, but l-Stepharanine(l-SPD) could not through the blood-brain barrier.5. The pharmacokinetics of l-Tetrahydrobeberruine(l-THR) in ratsObjective: To develop a LC-MS/MS method for the quantification of l-Tetrahydrobeberruine(l-THR) in rat blood, and study its pharmacokinetics and bioavailability in rats.Methods: 10 rats were randomly divided into 2 groups, one group was intravenous injected with l-Tetrahydrobeberruine(l-THR) 3 mg?kg-1, after administration 0、2、5、15、30 min and 1、2、4、8、12、24 h, retinal venous plexus blood, blood sample analysis preprocessing backward then set into LC-MS/MS to measure its concentration; another group was given l-Tetrahydrobeberruine(l-THR) 10 mg ? kg-1of oral administration, after administration 0、5、15、30 min and 1、2、4、8、12、24 h,retinal venous plexus blood, blood sample analysis preprocessing backward then set into LC-MS/MS measure its concentration.Results: The LC-MS/MS method we developed was simple、rapid、high sensitivity、good precision with little matrix effect. The calibration curve was liner across the concentration range of 0.5~2500 ng?m L-1 for l-Tetrahydrobeberruine(l-THR) with a lower LOQ of 0.5 ng?m L-1. The within-run and between-run precision(RSD%) was in the range-10.7%~10.9%. l-Tetrahydrobeberruine(l-THR) reached the peak concentration at 0.23 h after dosing. The main pharmacokinetic parameters were as follows : ig : t1/2(0.23±0.07)h,Cmax(1760.99±180.81)ng?m L-1,AUC0~24(397.03±97.39)ng?m L-1?h, AUC0~∞(397.77±97.92)ng?m L-1?h;iv:t1/2(4.25±2.25)h,Cmax(135.02±36.31)ng?m L-1,AUC0~24(409.35±90.64), AUC0~∞(617.42±167.60)ng?m L-1?h.