Studies On Preparation And Pharmacokinetics Of Sinomenine Hydrochloride PEGylated Transfersomes

Objective To prepare sinomenine hydrochloride(SIN-HCl)PEGylated transfersomes(SHPT)edge activated by volatile oils from traditional Chinese medicine with enhancing percutaneous absorption and synovial targeting of the encapsulated drug.To clarify the above characteristics by vehicle characterization,in vitro tests of percutaneous permeation and skin deposition and in vivo pharmacokinetics.Methods The HPLC method was selected to determine the content of SIN-HCl in SHPT.SHPT were prepared by ethanol injection method.After single factor experiments of the formulated quality of cholesterol,volatile oil and DSPE-PEG 2000,the formulation of SHPT were further optimized by an orthogonal design experiment.Morphology was observed by transmission electron microscope,mean particle size,polydispersity index(PDI)and Zeta potential were determined by laser scattering particle size analyzer,elasticity of the bilayers was evaluated by extrusion method,entrapment efficiency of SIN-HCl was determined by centrifugation-ultrafiltration method.The skin permeation and deposition tests were carried out with a Franz diffusion cell fitted with excised rat skin.The in vitro skin permeation and deposition characteristics of the optimal formulation SHPT(SHPT-P06V1/15)were determined and the effects amount of volatile oil and DSPE-PEG 2000 on skin permeation and deposition characteristics were investigated.Institute of Cancer Research(ICR)mice and rabbit were selected as experimental animals for the investigation of pharmacokinetic properties of SHPT-P06V1/15 after transdermal administration on the abdominal skin and the knee joint area,respectively,with sinomenine hydrochloride liposomes(SIN-HCl-LPSs)being used as control group.Microdialysis sampling technique was adopted in the pharmacokinetical study.The HPLC-MS/MS method was used for the determination of SIN-HCl in microdialysis samples.Results Compared with film dispersion method,ethanol injection formed SHPT with better flexibility.Single factor experiments showed that the effect of CH amount on the elasticity of SHPT were negligible.However,as the quantity of DSPE-PEG 2000 increased,the elasticity of SHPT increased first and then decreased with a maximal value.The quantity of volatile oil followed the same way.Orthogonal design experiment gave an optimal formulation with 18 mg of DSPE-PEG 2000 and 20 mg of volatile oils.The SHPT-P06V1/15 were mainly unilamellar vesicles with roundish shape.The mean particle size,PDI,Zeta potential,elasticity of bilayers and the entrapment efficiency of SIN-HCl were(109.10 ± 1.80)nm,(0.156 ± 0.007),(-18.90±2.21)mV,(24.50 ± 0.50)min and(22.38±0.79)%·The in vitro skin permeation experiments showed that the cumulative permeated drug quantity and percutaneous permeation rate of SHPT with 6%(as percentage of the weight of phosopholipids)of DSPE-PEG 2000 was prominently higher than those of SHPT with 2%,4%and 10%of DSPE-PEG 2000,respectively.However,the percutaneous permeation curves of the latter three formulations were not different obviously from each other.Similarly,the in vitro skin permeation and deposition experiments showed that the cumulative permeated drug quantity and percutaneous permeation rate of SHPT with 1/15(as fraction of the weight of phosopholipids)of volatile oils was prominently higher than SHPT with 1/20,1/8 and 1/6 of volatile oils,respectively.However,the percutaneous permeation curves of the latter three formulations were not different obviously from each other.The cumulative permeated drug quantity of SHPT-P06V1/15 was about 3 times of that of the liposomes(LPSs)and with a prominent larger percutaneous permeation rate compared with the latter.The in vitro skin deposition experiments showed that the amount of deposited drug in whole skin of the optimized formulation SHPT-P06V1/15 was higher than those of other SHPT and LPSs,but the amount of deposited drug in stratum corneumwas lower than those of other SHPT and LPSs.The area under the concentration-time curve(AUC0-t)and the Cmax of SHPT-P06V1/15 were 7.88-fold and 7.85-fold of those of SIN-HCl-LPSs,respectively,after application to the abdominal skin of ICR mice.The AUC0-t and Cmax of SHPT-P06V1/15 were 2.41-fold and 2.61-fold of SIN-HCl-LPSs,respectively,after application to the skin around the knee joint of rabbits.Conclusion The ethanol injection method is simple and reliable for the preparation of SHPT and the process has a good reproducibility.In vitro skin permeation and deposition tests,the SHPT with suitable quantity of DSPE-PEG 2000 and volatile oils from traditional Chinese medicine has prominently high skin permeation and increased skin deposition(mainly in deep skin).In vivo studies,SHPT has enhanced percutaneous absorption and synovial targeting ability.The results of our research provide a practical reference and theoretical basis for the development of transdermal targeting formulation based on PEGylated TFSs.