| Ginger is a kind of perennial herb with aromatic and pungent smell,which is widely distributed all over the world.Its rhizome is a kind of medicinal material,which can be used as medicine.6-shogaol(6-sh)is one of the main active components in ginger,and has many pharmacological effects,such as anti-tumor,anti-inflammatory,anti oxidant,anti atherosclerosis and so on.In recent years,the basic and clinical research of 6-sh has attracted the attention of scholars at home and abroad.Because of its low water solubility,pungent odor and low oral bioavailability,its clinical application is greatly limited.Therefore,in this paper,6-sh liposomes and Vitamin E polyethylene glycol succinate(TPGS)modified 6-sh liposomes were prepared respectively,so as to increase the solubility and oral bioavailability of 6-sh,and improve its antitumor effect.Chapter one: ReviewIn this chapter,the physical and chemical properties and pharmacological activities of 6-sh were reviewed,and the research progress of liposomes was introduced,including the advantages and disadvantages of different preparation methods.Finally,the physical and chemical properties of TPGS,the application of TPGS in pharmaceutics and the application of TPGS in anti-tumor were elaborated,which laid a solid theoretical foundation for the research of this paper.Chapter two: Pre-formulation study and extraction,purification and identification of 6-shogaolIn this chapter,ethyl acetate was used to extract gingerol,silica gel was used to roughly separate 6-sh,and C18 column with higher degree of separation was selected to purify the crude 6-sh,then the type and content of eluent were determined by TLC and HPLC,so as to obtain 6-sh purified product,and then the structure was identified by NMR,The equilibrium solubility of 6-sh in different media(HCl solution at p H 1.2,double distilled water,PBS buffer at p H 6.8,PBS buffer at p H 7.4)and the oil-water partition coefficient of 6-sh were determined.The purity of purified 6-sh was 97.45%,and the standard curve was y = 80962x-44176(R2 = 0.999,n = 3).The in vitro analytical method met the requirements of methodology.The equilibrium solubility of 6-sh in HCl solution at p H 1.2,double distilled water,PBS buffer at p H 6.8 and PBS buffer at p H 7.4 were 28.38 ± 2.04 μg/m L,23.99 ± 1.49 μg/m L,32.32 ± 2.47 μg/m L,34.75 ± 2.61 μg/m L respectively,and the log P is 2.87 ± 0.23.Chapter three: Preparation,formulation optimization and in vitro evaluation of 6-shogaolIn this chapter,the formulation of liposomes was screened and optimized.The effects of the following four factors on the particle size and entrapment efficiency of liposomes were investigated by single factor experiment: the ratio of drug to phospholipid,the amount of TPGS,the ratio of drug to sodium cholate,and the amount of IPM.Then the optimal formulation was screened by orthogonal design,and the optimal TPGS-6-sh-lip formulation was obtained as the ratio of drug to phospholipid 8:1,The dosage of TPGS was 5 mg,the ratio of drug to sodium cholate was 1:7,and the dosage of IPM was 63 mg.The particle size,PDI,zeta potential,entrapment efficiency and drug loading of TPGS-6-shlip prepared according to the formula were 23.50 ± 0.09 nm,0.140 ± 0.003,-38.52 ± 3.38 mv,95.18 ± 0.28 %,4.35 ± 0.02 %,The particle size,PDI,zeta potential,entrapment efficiency and drug loading of 6-sh-lip prepared without TPGS were 40.35 ± 3.71 nm,0.242 ± 0.036,-35.70 ± 3.16 mv,92.32 ± 0.32 %,4.29 ± 0.02 %,The results showed that tpgs-6-sh-lip could significantly increase the solubility of 6-sh compared with 6-sh,and also increase the solubility of 6-sh compared with 6-sh-lip.The stability of tpgs-6-sh-lip showed that tpgs-6-sh-lip had good stability in 30 days at different concentrations and temperatures,it is also more stable than 6-sh-lip.Chapter four: Pharmacokinetics and tissue distribution of 6-shogaol and its preparations in vivoIn this chapter,a method of HPLC analysis and detection for 6-sh in vivo with high specificity,high precision,good stability and high recovery rate was established.Meanwhile,the standard curve equation of 6-sh in rat plasma and different tissues and organs of mice was established.The drug metabolism and tissue distribution of 6-sh-lip in rats were investigated,The results of pharmacokinetics showed that T1/2 of TPGS-6-sh-lip was 1.26 and 1.87 times of that of 6-sh-lip and 6-sh;Cmax is 1.26 and 2.28 times of 6-sh-lip and 6-sh;Tmax is twice as high as that of 6-sh;The bioavailability(AUC)was 2.06 and 5.80 times of that of 6-sh-lip and 6-sh;MRT was 1.23 and 1.82 times of 6-sh-lip and 6-sh;The results of tissue distribution in mice showed that TPGS-6-shlip was less recognized by the reticuloendothelial system than 6-sh-lip,and decreased intake by liver and spleen.At the same time,the ability of 6-sh to enter brain tissue through blood-brain barrier could be increased due to the effect of TPGS.Chapter five: Antitumor activity of 6-shogaol and its preparationsIn this chapter,we investigated the effects of different concentrations(25,75,150,300 μg/m L)of 6-sh,6-sh-lip and TPGS-6-sh-lip on the proliferation of human hepatoma cell line Hep G2 at 24 h and 48 h in vitro.The results showed that the IC50 of 6-sh,6-sh-lip and TPGS-6-sh-lip on Hep G2 cells were 206.60 ± 8.83 μg/m L,206.45 ± 10.81 μg/m L,191.47 ± 6.34 μg/m L at 24 h,The IC50 of 6-sh,6-sh-lip and TPGS-6-sh-lip on Hep G2 cells were 178.98 ± 4.83 μg/m L,149.74 ± 5.16 μg/m L,132.27 ± 7.89 μg/m L at 48 h.Compared with 6-sh and 6-sh-lip,TPGS-6-sh-lip showed a better ability to inhibit the proliferation of tumor cells,and could significantly improve the anti-tumor effect of 6-sh. |
