Preparation Of Glycyrrhetinic Acid Complex TOGA-X4 Microparticles And Study Of In Vitro And In Vivo Correlatio

Objective:Provided ideas for preparation of the natural combined compound microparticles through preliminary study on the preparation technology of the TOGA-X4 glycyrrhetinic acid composition by stainless steel rapid membrane emulsification solvent volatilization method.Optimize the freeze-drying process of the microparticle freeze-dried powder through the investigation of the freeze-drying curve.Increase the dissolution rate and bioavailability of TOGA-X4 by reducing the particle size and increasing the specific surface area.Provide evidence for the development of new anti hepatoma drugs and explore the correlation between the particles in vivo and in vitro by study of release and pharmacokinetics.Method:Prepare TOGA-X4 particles by rapid membrane emulsification of stainless steel based on the preparation process of silybin PLGA microspheres,silybin particles and oleanolic acid particles.Select the best preparation process by using single factor analysis method and taking particle mean diameter,polydispersity index and rehydration as main indexes.Characterize the quality of the TOGA-X4 particles by study of the rehydration,crystallization and drug content uniformity.Optimize the optimum freeze-drying process through the investigation of the freeze-drying curve with the appearance and water rehydration as the index.Compare the release behavior of TOGA-X4 particles and TOGA-X4 in vitro by direct drug release.Compare the pharmacokinetic characteristics of TOGA-X4 particles and TOGA-X4 and discuss the correlation between in vivo and in vitro by drawing the drug time curve and tissue distribution.Results:1.The optimization of the preparation process is as follows:the oil concentration is 0.5%,the concentration of PVA is 3%,the oil to water ratio is 1:1,the membrane pressure is 0.4 MPa,the freeze-drying protective agent is 5%sucrose aqueous solution,the curing temperature is 70 degrees,and the curing solution are PVA(3%)and NaCl(0.45%).2.The optimization of the freeze-drying curve is as follows:put in at-20℃ rise from25℃ to 25℃ in 50 min,maintain 130 min,take out of the box after the lift test.Because of the restriction of laboratory conditions,the freeze drying curve was not used in later period.3.Physicochemical properties investigation results:morphology under scanning electron microscope is regular beads shaped.Water rehydration is excellent and the average particle size is(944.3 ± 81.3 0)nm,PDI is(0.29 ± 0.06).The results of DSC and XRD suggest that the crystalline form of the drug changes after the membrane emulsification.4.Content uniformity study results:the content determination method of HPLC was established,the column was Kromasil C-18 column(4.6×250 mm,5 μm),the mobile phase was methanol-water=95:5,the volume flow rate was 1 mL·min-1,the column temperature was 30 degrees,the detection wavelength was 254 nm,and the injection volume was 5μL.The content of TOGA-X4 in multiple batches was 36.78%,and the value of RSD was 2.29%.The results were qualified and the reproducibility of the batch was good.5.In vitro release results:1%SDS pH=6.5(release medium Ⅰ)and pH=7.4(release medium Ⅱ)phosphate buffer salt solution were selected as release mediums.In I release medium,the cumulative release rate of TOGA-X4 particles of 72 h was up to 98.88%,the release of model fitting results:Logstic equation>first order equation>Higuchi equation>zero order equation;Cumulative release rate of TOGA-X4 powder of 72 h was only 16.65%,the equation fitting results:zero order equation>Logstic equation>Higuchi equation>first order equation.In the release medium Ⅱ:cumulative release rate of particles of 72 h was 94.75%,the release model fitting degree was as following:Logstic equation>Higuchi equation>First order equation>Zero order equation;TOGA-X4’ was only 15.67%,model fitting degree was as following:zero order equation>Logstic equation>first order equation>Higuchi equation.6.In vivo pharmacokinetics results:the plasma concentration of TOGA-X4 reached to peak two times,the first peak is 6 h and the average blood concentration was 0.3972 μg·mL-1,the second peak is 12 h,the average blood concentration was 0.4456 μg·mL-1.The results of tissue distribution showed that the concentration and content of the microparticles in liver tissue were high and had strong liver targeting.Conclusion:Microparticles prepared by stainless steel membrane emulsification is simple,stable and reliable.It improves the dissolution of insoluble drugs and increases the insoluble drug administration.Stainless steel membrane emulsification has superiority in preparation of insoluble drug particles and provides a new idea for the development of new anticancer preparation.