Study On BCS Classification And Absorption Mechanism Of Effective Components Of Chinese Herbs

Objective:Terpenoids are widely distributed in nature and have many biological activities such as anti-oxidation,anti-inflammatory,immune enhancement,anti-malaria and anti-cancer and so on.Oral dosage forms are widely used,but currently there are few oral dosage forms for terpenoids,which may be a problem of low oral bioavailability of terpenoids.The Biopharmaceutics classification system(BCS)is an evaluation method for the bioavailability of oral drugs based on the in vitro solubility and intestinal permeability of drugs.The composition of traditional Chinese medicine is complicated,and there is no systematic method for comparing the biopharmaceutics classification of traditional Chinese medicine.Therefore,this experiment mainly compares eight kinds of terpenoid monomers,such as Paeoniflorin,Linalool,Artemisinin,Artesunate,Ginkgolide A,Stevioside,Curcurbitacin Ila and Betulinic acid as model drugs.A systematic and comprehensive approach to determine the BCS classification of drugs and their absorption mechanisms.Method:Firstly,using MOE,Discovery Studio and StarDrop software,eight kinds of terpenoid monomers,such as Paeoniflorin,Linalool,Artemisinin,Artesunate,Ginkgolide A,Stevioside,Curcurbitacin 11a,Betulinic acid predictions were made to determine its solubility,extent of intestinal absorption,and whether it was regulated by P-glycoprotein(P-gp).The solubility of these eight terpenoids was then tested according to the guidelines of the FDA-defined biopharmaceutical classification and the solubility method in the 2015 Chinese Pharmacopoeia.The HPLC and UPLC methods were established to determine the content of the test compound.The excess test compound was dissolved in pHl.2,4.0,6.8 phosphate buffer and water,and shaken at 37? for 3h under shaking conditions.The supernatant was subjected to liquid phase detection to calculate the DO value and the dissolved content in different buffers to determine the solubility.The in vitro cell Caco-2 cell model was established to determine the permeability and transport mechanism of these eight terpenoids.The MTT method was used to select the concentration of the drug in two-way transport experiments,and the low,medium and high doses with drug survival rate higher than 90%were selected.Experiments were performed at each concentration.At the same time,the artificial permeability membrane model PAMPA was established to further verify its permeability,the high,medium and low drug concentrations were selected for permeability experiments.The permeability was judged based on its apparent permeability coefficient(Papp)and effective permeability(Pe)and the effect of transit time,drug concentration,and P-gp inhibitor on drug transport.Result:The software predicts that MOE indicates that Paeoniflorin,Linalool,Artemisinin,Artesunate,and Ginkgolide A have good water solubility,stevioside and Curcurbitacin IIa are poorly water-soluble,and Betulinic acid is very poor in water solubility.The software Discovery Studio showed thatPaeoniflorin,Linalool,Ginkgolides A,andCurcurbitacin IIa were well-soluble in water,Artemisinin,Artesunate,andStevioside were poorly water-soluble,and Betulinic acid was very poor in water solubility.Paeoniflorin and ginkgolides A have poor intestinal absorption properties;The absorption of the Curcurbitacin ?a is moderate;Linalool,Artemisinin,Artesunate and Stevioside have good intestinal absorption properties;The absorption properties of Betulinic acid intestines are better.StarDrop software predicts the solubility of Paeoniflorin to be slightly soluble,the solubility of Linalool is slightly soluble,and the solubility of Artemisinin,Ginkgolides A,Artesunate,Stevioside,Curcurbitacin ?a and Betulinic acid is extremely soluble.The drug absorption of Paeoniflorin,Artesunate and Curcurbitacin ?a is less than 30%,and the others are more than 30%.Both are not regulated by P-glycoprotein(P-gp).According to the US FDA guidelines,the D0 value of the drug is greater than 1 except for Paeoniflorin,indicating that Paeoniflorin is a highly soluble drug,and the others are low-solubility drugs.In the solubility standard of the Chinese Pharmacopoeia in 2015,except Paeoniflorin other all is a low-solubility drug.In the Caco-2 cell monolayer model,the Papp values of the AP-BL and BL-AP sides of Paeoniflorin,Linalool and Ginkgolides A were less than14.96×10-6 cm·s-1 which was a low-permeability drug;The Papp values of Artemisinin,Artesunate,Stevioside,Curcurbitacin Ila and Betulinic acid on the AP-BL and BL-AP sides were all greater than14.96×10-6 cm·s-1,which was a highly permeable drug;Except for Linalool and Curcurbitacin IIa,other six test drugs the ER values were less thanl.5 mainly passive transport.Add verapamil to the transport experiment,there was no significant change in the transport rate of Ginkgolides A.The transport rate of Linalool,Artemisinin and Curcurbitacin ?a slows down,It indicates that its transport is not regulated by P-gp,not a substrate for P-gp.The transport rates of the other four tested drugs were significantly faster,indicating that their transport was regulated by P-gp and was a substrate for P-gp.In the PAMPA model,Paeoniflorin,Linalool and Ginkgolides A have a Pe value of less than 1×10-6 cm.s-1,which is a low permeability drug;Artemisinin,Artesunate,Stevioside,Curcurbitacin ?a and Betulinic acid have Pe values greater than 1×10-6 cm·s-1,which are highly permeable drugs.Conclusion:Artemisinin,Artesunate,Stevioside,Curcurbitacin ?a and Betulinic acid are BCSII drugs,Linalool and Ginkgolides A are BCSIV drugs,Paeoniflorin is a BCS class III drug.The transport mode of paeoniflorin,artemisinin,artesunate,ginkgolides A,stevioside,and betulinic acid is mainly passive transport;The transport mode of Curcurbitacin ?a and Linalool is based on active transport.Paeoniflorin,Artesunate,Stevioside and Betulinic acid are regulated by P-gp and are P-gp substrates.Linalool,Artemisinin,Ginkgolides A and Curcurbitacin ?a are not regulated by P-gp,not P-gp substrates.