局部应用的辣椒碱纳米脂质载体的制备、表征及其体内外评价

发布时间：2018-05-31 01:12

本文选题：辣椒碱 + Box-Becken设计　；参考：《浙江大学》2017年硕士论文

【摘要】：目的：本研究拟构建辣椒碱纳米脂质载体局部给药系统,通过单因素分析以及Box-Becken设计优化处方,以期提高辣椒碱的透皮量,能够产生镇痛、抗炎作用,同时减少辣椒碱局部应用的皮肤刺激性。方法：1.通过单因素分析筛选影响辣椒碱纳米脂质载体包封率和粒径的处方因素；2.利用Box-Becken设计分析不同处方因素对响应值的影响,模拟得到优化处方,对合成的辣椒碱纳米脂质载体进行物理化学性质表征并考察体外释放情况；3.通过体外细胞实验,考察辣椒碱游离药物和辣椒碱纳米脂质载体对皮肤细胞的毒性；4.通过体外经皮渗透实验,考察辣椒碱制剂的透皮效果和不同皮层内辣椒碱的滞留量；考察纳米脂质载体促进药物透皮的深度和主要途径；5.采用热板法考察辣椒碱制剂对ICR小鼠的镇痛作用；采用角叉菜胶大鼠足趾肿胀模型考察辣椒碱制剂的抗炎作用,并探究其抗炎机制；6.考察不同浓度辣椒碱制剂的皮肤刺激性；7.通过筛选适宜冻干保护剂,制备辣椒碱纳米脂质载体冻干粉剂。结果：利用Box-Becken设计优化处方,考察固液脂质比例、脂质投入量和表面活性剂使用量对包封率和粒径的综合影响,得到最优处方为：脂质总质量为0.86g,其中固体脂质占58%；表面活性剂用量为3.6%。制备得到的辣椒碱纳米脂质载体溶液呈乳白色且带有乳光,粒径均一,约为119nm,形态圆整,包封率为91.6%且在体外释放48h过程中具有良好的缓释效果。MTT实验和细胞周期实验证明纳米脂质载体能够降低辣椒碱对皮肤细胞的毒性,增加使用的安全性。体外24 h经皮渗透实验后,辣椒碱纳米脂质载体及其凝胶的单位面积累积透过量分别为29.7μg.cm-2和25.6μg. cm-2,较辣椒碱市售软膏组和辣椒碱溶液组的17.9 μg. cm-2和2.2 μg. cm-2显著提高；辣椒碱纳米脂质载体及其凝胶的透皮速率较辣椒碱溶液组分别提高14.4倍和12.2倍,表明纳米脂质载体能够显著提高辣椒碱的透皮效率。通过检测辣椒碱的皮内滞留量证明纳米脂质载体能够显著增加辣椒碱在表皮和真皮中的滞留。利用细胞膜绿色荧光探针(3,3'-dioctadecyloxacarbocyanine perchlorate, Dio)代替辣椒碱进行药物透皮深度和主要透皮途径的考察,结果发现Dio溶液组大约渗透至150μm皮肤深度,而Dio纳米脂质载体凝胶组和Dio纳米脂质载体组可渗透至皮肤内约210μm和260μm,结合皮肤切片观察结果表明纳米脂质载体主要是通过毛囊附属器途径促进药物透皮,增加药物的透皮深度。体内实验结果表明辣椒碱纳米脂质载体具有镇痛作用且呈剂量依赖性并可以通过抑制PGE2信号通路产生抗炎作用。皮肤刺激性实验结果显示纳米脂质载体能够减轻辣椒碱的刺激性。利用筛选得到的冻干保护剂：2%蔗糖+2%葡萄糖制备辣椒碱纳米脂质载体冻干粉剂,发现其具有较好的复溶性且复溶后纳米粒形态并未发生明显变化。结论：经Box-Becken设计优化处方后制备得到的辣椒碱纳米脂质载体溶液及其凝胶具有良好的体外透皮效果,能够在体内产生镇痛和抗炎作用的同时减少辣椒碱的刺激性。
[Abstract]:Objective: to construct the local drug delivery system of capsaicin nano lipid carrier, and optimize the prescription by single factor analysis and Box-Becken design in order to improve the transdermal volume of capsaicin, to produce analgesic and anti inflammatory effects, and to reduce the skin irritation of capsaicin local application. Method: 1. screening of peppers by single factor analysis. The formulation factors of encapsulation efficiency and particle size of alkali nano lipid carrier were analyzed. 2. Box-Becken was used to analyze the effect of different prescription factors on the response value. The optimized prescription was simulated. The physicochemical properties of capsaicin nanoparticles were characterized and released in vitro. 3. through in vitro cell experiments, capsaicin dissociation was investigated. The toxicity of drugs and capsaicin nanoparticles on skin cells; 4. through the skin penetration test in vitro, the transdermal effect of capsaicin preparation and the retention of capsaicin in different cortex were investigated, and the depth and main ways of promoting drug penetration by nano lipid carriers were investigated. 5. the hot plate method was used to investigate the town of capsaicin on ICR mice. Pain effect; using the rat toe swelling model of carrageenin to investigate the anti-inflammatory effect of capsaicin and explore its anti-inflammatory mechanism; 6. to investigate the skin irritation of capsaicin in different concentrations; 7. to prepare capsaicin nano liposome freeze-dried powder by screening suitable freeze-drying protective agent. Results: the optimized prescription of Box-Becken was designed by using the formula. The optimum prescription of lipid ratio, lipid input and use of surfactant on encapsulation efficiency and particle size was obtained: the total mass of lipid was 0.86g, in which the solid lipid accounted for 58%, and the capsaicin nano lipid carrier prepared by the dosage of surfactant was milk white with milk light, and the particle size was uniform. About 119nm, round and round, the encapsulation rate is 91.6% and has good release effect during the release of 48h in vitro..MTT experiment and cell cycle experiment prove that nano lipid carrier can reduce the toxicity of capsaicin to skin cells and increase the safety of use. After 24 h transdermal permeation test, capsaicin nano lipid carrier and its gel The cumulative overdose of the unit area was 29.7 mu g.cm-2 and 25.6 g. cm-2 respectively. The 17.9 Mu g. cm-2 and 2.2 u g. cm-2 of capsaicin group and capsaicin solution group were significantly improved, and the transdermal rate of capsaicin nanoparticles and its gel were 14.4 and 12.2 times higher than that of capsaicin solution group respectively. The transdermal efficiency of capsaicin is improved. The retention of capsaicin in the epidermis and dermis can be significantly increased by detecting the intradermal retention of capsaicin. The penetration depth and main transdermal pathway of the drug using the cell membrane green fluorescent probe (3,3'-dioctadecyloxacarbocyanine perchlorate, Dio) instead of rimperine The results showed that the Dio solution group penetrated to the skin depth of about 150 mu m, and the Dio nano lipid carrier gel group and the Dio nano lipid carrier group permeated into the skin about 210 mu m and 260 micron m. The results of skin slice observation showed that the nano lipid carrier was mainly through the hair follicle accessory pathway to promote the drug transdermal and to increase the drug transdermal delivery. The results of in vivo experiments showed that capsaicin nanoparticles had analgesic and dose-dependent effects and could inhibit the anti-inflammatory effect by inhibiting the PGE2 signaling pathway. Skin irritation results showed that nano lipid carriers could reduce capsaicin irritation. The screened lyophilized protective agent, 2% sucrose +2% glucose, was used. The lyophilized powder of capsaicin nano lipid carrier was prepared. It was found that it had good resolubility and the morphology of the nanoparticles did not change obviously. Conclusion: the capsaicin nano lipid carrier solution and its gel prepared after the optimized formulation of Box-Becken and its gel have good external transdermal effect and can produce analgesic and analgesic effects in the body. The anti inflammatory effect reduces the irritation of capsaicin.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943

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