The Pharmacokinetic Characteristics Of CQ Formula In Rats

The pharmacodynamics research will lead to on the basis of RP-HPLC method is used to simultaneously determine indexes component sinomenine hydrochloride and ligustrazine hydrochloride blood drug concentration,the CQ formula ingredient sinomenine hydrochloride and ligustrazine hydrochloride pharmacokinetic characteristics;And through the investigation of CQ formula in normal rats and bone cancer pain model in rat plasma pharmacokinetic process,more normal and pathological condition of pain medicine generation of dynamic characteristics of difference,for the clinical application of CQ formula in the treatment of pain to provide more valuable information for reference;To clarify the interaction between the main components of CQ formula,sinomenine hydrochloride and ligustrazine hydrochloride,to provide the basis for revealing the mechanism of CQ formula action and clinical drug delivery.Conducting the method for determing Sinomenine HCL and Ligustrazine HCL concentration in plasma by RP-HPLC.Sinomenine HCL and Ligustrazine HCL concent in plasma was determined by internal standard method,using theophylline as an internal standard.HPLC conditions for analyzing Sinomenine HCL and Ligustrazine HCL in plasma samples were:the chromatographic column Extension-C₁₈?4.6 mm x 250 mm,5?m,Agilent,USA?.Mobile phase A:phosphate buffer saline?adjusting pH to 9.4⁹.5 with trithylamine?,mobile phase B:100%Methanol,gradient elute?0min,40%B;4min,45%B;12min,55%B;15min,40%B,v/v?;flow rate of 0.8 mL·min^-1,column temperature:30?,UV detection wavelength:265nm?280nm.By the method validation,the precision,accuracy,stability and recovery rate of the method are in accordance with the relevant regulations.The Sinomenine HCL in the plasma showed a good linear relationship with the peak area in concentration range from 0.05¹0?g·mL^-1,y=0.2914x-0.0099?R²=0.9978?,take(10?g·mL^-1?1.25?g·mL^-1?0.625?g·mL^-1)three concentration measurement extraction recovery rate was 100.08%,99.51%,104.94%,the within-rin for rat piasma of Sinomenine HCL was 3.25%,3.59%,3.92%;the between-run precision for rat piasma of Sinomenine HCL was 3.98%,6.74%,5.72%.The stability for rat piasma of Sinomenine HCL in 8h was 9.98%,7.72%,4.46%.The Ligustrazine HCL in the plasma showed a good linear relationship with the peak area in concentration range from 0.025¹0?g·mL^-1,y=0.8615x+0.169?R²=0.9985?,take(10?g·mL^-1?1.25?g·mL^-1?0.625?g·mL^-1)three concentration measurement extraction recovery rate was 99.67%,100.43%,97.85%,the within-rin for rat piasma of Ligustrazine HCL was 3.66%,3.83%,6.49%;the between-run precision for rat piasma of Ligustrazine HCL was 4.13%,5.08%,5.23%.The stability for rat piasma of Ligustrazine HCL in 8h was 10.27%,7.27%,5.01%.The RP-HPLC method for determining Sinomenine HCL and Ligustrazine HCL in rat plasma was simple,accurate,less interference,high sensitivity and strong specificity;endogenous impurities were removed with samples preparation;They were suitable in physiological and pathological state in rats plasma drug concentration determination and pharmacokinetic study.To establish metastatic cancer pain of tibia rats model.Screen out qualified adaptability stimulation rats were divided into control group and model group,take 5uL Walker 256 ascites cancer cells injected into rat tibia bone marrow cavity,Von Frey hairs to evaluate mechanical pain threshold of rats.Cause the model rats give CQ formula high dose(40mg·kg^-1),records before and after the drug 0min,30min,60min,90min,120min,180min,240min,360min,when mechanical pain threshold,evaluation of CQ formula in rats pharmacodynamics action bodily.Building 9¹1days after stimulation rats foot to evaluate mechanical pain threshold,the mechanical pain threshold for three consecutive days will be 8g as a model or less successful,metastatic cancer pain of metastatic cancer pain of tibia rats threshold by preoperative?22.85±5.36?g to?6.57±2.22?g,the postoperative period of 9¹2 days was maintained at?6.57±2.22?g^6.85±1.57g,and there was no significant change before and after the operation of the sham group;the analgesic effect was significant in 30min¹80min.The results showed that CQ formula had an interventional effect on metastatic cancer pain of tibia rats,and the pharmacodynamic behavior of rats with postoperative bone cancer was similar to that of pharmacokinetics.Therefore,in this paper,the CQ formula in normal and metastatic cancer pain of tibia rats models pharmacokinetic characteristics were discussed,and analyzed the CQ formula ingredient the rationality of sinomenine hydrochloride and the ligustrazine hydrochloride compatibility,understand the blood drug concentration change rule,to reveal the CQ formula mechanism of action and provide basis for clinical regimen.Based on the established RP-HPLC method,compared the differences of pharmacokinetic characteristics in plasma of normal rats and metatarsal cancer pain model rats.After feeding adaptability rats were randomly divided into CQ formula high group(40mg·kg^-1),CQ formula medium group(20mg·kg^-1),CQ formula low group(10mg·kg^-1),sinomenine hydrochloride(40mg·kg^-1)and ligustrazine hydrochloride group(40mg·kg^-1),the application of fully automatic blood instrument in animals at 0min,5min,15min,30min,45min,60min,90min,120min,180min,240min,360min,480min,720min,1440min continuous blood in time.The results showed that CQ formula had a certain difference in pharmacokinetics in rats with model rats and normal rats,and the pathologic state of bone cancer affected the metabolism kinetics of CQ formula.Compared with normal rats,CQ formula in model group rats plasma sinomenine hydrochloride in compound and ligustrazine hydrochloride area under the plasma concentration curve were decline,respectively?1320.462±354.313⁴68.196±85.290?,?3811.300±2889.793¹978.690±821.924?;Up to reduction in the peak concentration Cmax,respectively?10.278±2.655¹.938±0.189?,?22.481±1.253¹0.203±1.267?,plasma clearance speed?CL_z/F??0.031±0.009⁰.087±0.016?,?0.014±0.006⁰.023±0.009?,indicated that CQ formula under the condition of metastatic cancer pain pathology absorption accelerated,total clearance rate is accelerated,metastatic cancer pain pathological conditions may change CQ pharmacokinetic process of compound.To observe the metabolism of CQ formula in normal rats.Normal rats chloral hydrate anesthesia,jugular vein and femoral vein inserted into the blood vessels,wear vest fixation,postoperative fasting 12h after the second day early morning to medicine through femoral vein pipe according to the weight of CQ formula high(40mg·kg^-1),medium(20mg·kg^-1),low dose(10mg·kg^-1),volume 1mg·mL^-1,respectively before and after drug 0min,5min,15min,30min,45min,60min,90min,120min,180min,240min,360min,480min,720min,1440min jugular vein blood.The results showed that the AUC?0-t?at different time in the low dose of CQ formula was?1320.462±354.313?,?1460.592±815.747?,?55.036±92.344?,and C_max was?10.278±2.655?,?7.609±5.654?,?1.884±0.492?.Ligustrazine hydrochloride in CQ formula including in different time of AUC?0-t?,respectively?3811.300±2889.793?,?3911.579±776.192?,?83.248±713.548?and C_max respectively?22.481±1.253?,?11.395±1.544?,?5.091±1.182?,indicated that CQ formula blood drug concentration in dose dependent,decreased with the decrease of dosage and time.Comparison of sinomenine hydrochloride alone and CQ formula for medicine,found that sinomenine hydrochloride group and CQ formula high dose group of the MRT?0-t?,respectively?109.782±24.886?,?264.480±104.511?;t1/2z respectively?84.103±28.204?,?212.037±49.991?;V_z/F respectively?3.950±1.173?,?9.164±2.341?;T_maxax respectively?5±0?and?9±5.477?,clarify that ligustrazine hydrochloride can extend the time of sinomenine hydrochloride selfly,increasing the apparent volume of distribution,make its can rapid distribution and reduce exposure quantity,prolong the half-life of sinomenine hydrochloride and its T_max in vivo.Sinomenine hydrochloride and ligustrazine hydrochloride Cmax respectively?13.071±1.091?,?10.278±2.655?;AUC_?0-t?respectively?1207.511±170.577?,?1320.462±354.313?;CL_z/F respectively?0.034±0.005?,?0.030±0.009?;ligustrazine hydrochloride can prolong sinomenine hydrochloride residence time in vivo at the same time reduce the concentration of sinomenine hydrochloride,and the influence on the degree of absorption and clearance does not produce.Comparison of ligustrazine hydrochloride alone for medicine and the doses of CQ formula for medicine,found that ligustrazine hydrochloride and CQ formula high dose group of AUC?0-t?respectively?2100.059±313.204?,?3811.300±2889.793?;C_max respectively?10.053±1.396?,?22.481±1.253?;T_max respectively?12±10.954?and?9±5.477?;MRT_?0-t?respectively?191.250±48.867?,?111.248±133.563?;illustrate sinomenine hydrochloride can enhance the absorption of ligustrazine hydrochloride in vivo,the enhancement of the peak concentration,shorten the T_max and no influence on the retention time.V_z/F respectively?3.264±1.440?,?1.556±0.468?;CLz/F respectively?0.019±0.003?,?0.014±0.006?;illustrate sinomenine hydrochloride has little affect the condition of residence time reduces the distribution apparent volume of the ligustrazine hydrochloride and plasma clearance,slowing its diffusion length and removal rate,to achieve long-term,sustained release effects.The experimental results show that the CQ formula complex metabolic processes,to reflect the true metabolism of the drug,we will depend on in the normal rats and model rats pharmacokinetic process.So we studies the CQ formula in normal and metastatic cancer pain pharmacokinetics in rats,we will proved the rationality of the compound compatibility,provide the basis for guiding clinical medication.