Preparation Of The Triptolide Dry Emulsion And Study Of Its Antitumor Activities

Triptolide(TPL) separates from the Celastraceae plant Tripterygium Tripterygium(Tripterygium wilfordii Hook F.),which is one of the main active ingredient.Since TPL is poorly soluble in water, limiting its clinical use.It need to use an organic solvent such as propylene glycol co-solvent, in order to avoid the toxicity of co-solvent, reduce the stimulation of TPL on vascular endothelial cells,simultaneously in order to solve the problem of drug solubility and improve its stability, reduce the adverse toxicity, especially be caused phlebitis by intravenous injection, the subject proposed to study of TPL lipid emulsion. This paper was developed TPL dry emulsion: effects of prescription, preparation and formulation properties of emulsion, and the triptolide emulsion safety and preliminary study of anti-tumor efficacy and rat pharmacokinetics.Chapter I:the preparation of triptolide lipid emulsion and its dry emulsionIt is successfully established the HPLC method of TPL to determine the equilibrium solubility of the drug in different medium, soybean oil and medium chain fatty acid glycerides: a mixture of 1:3 as the oil phase, which preparing triptolide lipid emulsion by speed dispersion-high pressure homogenization. In the single factor determined which based on the prescription process: the amount of poloxamer188,egg yolk lecithin E 80 and oil for injection were 2,1.2,10(g/100mL);High-speed dispersion time was 90 s,high pressure homogenization pressure was 800 bar,homogenization times was 6 times. Its emulsion form prepared for the regular round,particle size was(190.0±6.0) nm, Zeta potential was(-14.6±4.3) mVP.I, value was 0.05± 0.06,drug content was(60.16±0.30) ?g/m L,encapsulation efficiency was(90.94±0.49)%.In vitro release of certain sustained release,in 240 minTPL lipid emulsion cumulative released of 95%.To solve the problem of instability in the emulsion during storage, further prepared dry emulsion by freeze-drying:TPL dry emulsion of 5% trehalose made after its dry emulsion; Reconstituted by adding an appropriate solvent, its system for particle size was(396.5 ± 11.4) nm, Zeta potential was(-22.9 ± 4.0) mV, P.I was 0.6 ± 0.02, water content was(0.05 ± 0.03)%.Chapter II: study on pharmacokinetic of TPL dry emulsion in vivoDosing 100?g/kg through tail vein injection with TPL and TPL dry emulsion, in vivo pharmacokinetics of the rat, researching on effects of different forms pharmacokinetic behavior in rats. By Das software for processing,TPL dry emulsion in rats maximum plasma concentration(Cmax) was 1.38 times than TPL injection,in vivo retention time(MRT0-180min) extended 2.33 times,the area under the curve(AUC0-180min) increased 2.75 times,and these results were statistically significant differences(* P<0.01).Thus,TPL dry emulsion in the body had obvious advantages,making the drug in the body a long time to maintain a high concentration of the drug and significantly prolonged in vivo circulation time.Chapter III: study on safety and anti-tumor effect of TPL dry emulsionAs an indicator to evaluate the hemolytic safety of triptolide dry emulsion,the results showed that TPL dry emulsion was non-hemolytic on rabbit vascular irritation and safe.In MTT assay TPL injection and dry emulsion on SGC-7901 andSW620,the proliferation of human tumor cell line was a concentration-dependent inhibition of tumor cell proliferation.In human colon carcinoma cell SW620 of nude mice xenograft model to evaluate TPL dry emulsion in vivo anti-tumor growth activity,inhibition rate respectively of 5-fluorouracil group,TPL injection group,TPL dry emulsion group on human colon cancer cells SW620 of nude mice xenograft model,were 54.68%,71.00%,80.00%,compared with the control group,P were less than 0.01.Conclusion:1. TPL made of lipid emulsion and dry emulsion stability were good.2.The pharmacokinetics study showed that the TPL group had markedly shorter MRT than the TPL-E group.The MRT of the TPL-E group were 2.33 times longer than that of.MRT0-180 min,t1/2 and AUC0-180 min of rats were significantly differences between injection and dry emulsion in the body, which was obvious advantages.3. The results showed that TPL dry emulsion acted non-hemolytic test on rabbit vascular irritation, which was good safety.4. TPL dry emulsion can inhibit a variety of in vitro proliferation of tumor cells; in vivo can strongly inhibit the tumor growth on human colon cancer SW620 cells of nude mice xenograft model, and inhibition rate greater than 5-FU group, but the toxicity was significantly lower than 5-FU group.