Study On The Preparation Process And Pharmacokinetics Of Capillipes A

The plant named Lysimachia capillipes Hemsl.often used in the treatment of malignant tumor,belongs to Lysimachia of Primulaceae.With heat-clearing,dispelling the wind,treating cough,regulating menstruation,calming and other effects,the drug commonly used in treatment of cold,cough,rheumatism,irregular menstruation,neurasthenia and malignant tumors.Its chemical constituents are mainly flavonoids and saponins.Preliminary study of research group found that the effective fraction to cure cancer is the total saponins.Capilliposide A,capilliposide B and capilliposide C are the effective components in total saponins with a significantly anti-tumor effect.Which Capilliposide-B and Capilliposide-C have strong hemolytic,it’s difficulty to prepared them into a single drug.But Capilliposide-A has poor hemolytic,anti-tumor effect is also strong,so it has a relatively large use of research value.Therefore,Capilliposide-A is the study object in this article,and its preparation technology and pharmacokinetics are investigated.First,the Capilliposide-A standard was prepared by hydrolysis.The Capilliposide-B monomer was hydrolyzed by sodium hydroxide and purified by solid phase extraction(SPE)to obtain the monomer compound.The product was identified by mass spectrometry and nuclear magnetic resonance.The hydrolyzate was Capilliposide-A.The area normalization method was characterized by high performance liquid-Evaporative light scattering detector is calibrated to determine the content of more than 98%.Second,to study the hydrolysis technology of total saponins,we established a method using HPLC-ELSD to detect the content of hydrolysate,which took Capilliposide A as index compounds.The hydrolyze process was screened by combination of single factor experiments and orthogonal design.The influences which caused by the kinds of the catalyst,phenol and time,temperature and etc were discussed.The optimum hydrolysis process was as follows:the total saponification ratio was 1 g:40 mL and the catalyst was 0.08 mol·L-1 NaOH aqueous solution.The hydrolysis temperature was 50℃ and the hydrolysis time was 144 h.The extraction rate were 39.77%in the optimized extraction method.To study the purification method of total saponins hydrolysate,static adsorption and desorption test were used to select suitable macroporous resin.Several factors include sample concentration,loading rate and diameter-height ratio were evaluated,the final purification process was optimized by dynamic test and orthogonal design of sample loading and eluent,with the combination of yield and purity of capilliposide A as composite index,The optimum conditions were as follows:loading concentration 20 g ·L-1,loading amount 145 mg·g-1,repeated loading 3 times,adsorption 0.5 h,loading rate 2 BV·H-1,water elution 3 BV,40%ethanol elution 3 BV,70%ethanol 4 BV,flow rate 2 BV·H-1.After purification according to the optimum procedure,the LC-A content was 63.25%,the transfer rate was 71.36%.Using the single factor and orthogonal test,the elution volume,loading volume and concentration and elution rate were investigated by LC-A purity and content.The optimum medium pressure reversed-phase column The sample volume was 0.006 BV,the loading concentration was 0.1 g·mL-1,the elution flow rate was 8 BV·H-1,and the elution volume was 14～17 BV.Purification was carried out according to the optimum process,and the purity of LC-A monomer was over 98%and the yield was 86.54%.Finally,to study the absorption and excretion properties of capilliside A,a reliable LC-MS/MS method was developed in rat plasma,urine and fecal waste.Results of pharmacokinetics in rats show there was a very low exposure after oral administration.Capilliside A in 5～15 mg·kg-1 intravenous dose range in rats showed dose-dependent pharmacokinetic characteristics;LC-A in urine and feces excretion is not high,the total excretion rate of 15.64%.In summary,capilliside A has a strong anti-tumor activity,the monomer drug has a large development,the monomer preparation process for the subsequent mass production to provide the basis for some pharmacokinetics of the study for its dosage form selection and drug application to provide a scientific basis.