Study On Derivatization And Pharmacokinetics Of Active Ingredients Of Fomitopsis Pinicola

Fomitopsis pinicola is used as medicinal “Sanghuang” in Northeast China folk.“Sang huang” is known as one of the large medicinal fungus with the best anti-cancer effect recognized internationally.It has been used as a traditional Chinese medicine for over two thousand years.Tsugaric acid A and pinicolic acid A are the main antitumor components of Fomitopsis pinicola.In this paper,combined with previous studies,the preparation process of two active components and pharmacokinetics was given.The study on stability,preliminary derivatization and toxicology tests of tsugaric acid A were presented.The research aims to complete part of pre-clinical research,lay foundation for the development of new drugs of traditional Chinese medicine.In this paper,solvent extraction,solid-liquid extraction,silica gel column chromatography and recrystallization were used to optimize the preparation process of active ingredients tsugaric acid A and pinicolic acid A.Compared with the first generation of active ingredient preparation,the yield of tsugaric acid A was 0.1087%,increased by 11.3 times,and pinicolic acid A was 0.0691%,increased by 7.9 times.In the study of the stability of tsugaric acid A,the results of influencing factors showed that:according to the sensitivity of drugs to various factors,temperature> humidity> light.It was stable in ethyl acetate solution and methanol solution,it was extremely unstable in chloroform solution and can be decomposed into 5 compounds,three of which were tsugaric acid A,trametenolic acid B and eburicoic acid.In short-time test,when the test conditions were: 25℃±2℃/relative humidity of 60%±10%,there were 2 batches of drugs significant changes within 3 months,there was no significant changes in 3 batches of drugs within 6 months.The long-run test obtain storage conditions were as follows: Not suitable for room temperature preservation,cryopreservation(-20℃±5℃),dry,obturation,protect form light and stay away from chloroform solution.In order to improve the polarity and solubility of tsugaric acid A,the preliminary derivatization of it was studied.After the glucoside was protected by benzyl,a new compound was synthesized by EDCl/DMAP condensation method of Steglich esterification.After identification of 13C-NMR,1H-NMR,MS,HSQC and HMBC,concluded that the new compound was tsugaric acid A benzyl glucose ester.The pharmacokinetics of two active ingredients were studied by the combination of internal standard method and HPLC method.Results show that,4 compounds were detected in rat plasma after intragastric administration,the excretion pathway of tsugaric acid A was digestive system,prototype drugs can be detected in feces.The results of pinicolic acid A after intragastric administration were analyzed by "3P97" software,accord with two compartment model.The maximum plasma concentration was reached at 45 min,and the higher the dose the higher the concentration of the drug in the body.From the apparent volume distribution(V/F)and apparent clearance of rate(CL/F),the distribution and excretion of drugs in rats were very fast.Acute toxicity of tsugaric acid A was tested by up-and-down procedure(UDP)method.The results of the experiment with“AOT425StatPgm” software analysis.It was concluded that the toxicity grade was low,LD50 was 1750mg/kg,95% confidence interval was 759.4~3090 mg/kg,Combined with the observation index of acute toxicity test,toxic effects may be due to the stimulation of respiratory neurons caused by cardiopulmonary insufficiency and pulmonary edema,and inhibit the central nervous system.