白复凝的制备及初步评价

发布时间：2018-09-03 13:45

【摘要】：白复凝是以渭北野生羊角蔓中抗白癜风有效部位为功效成分制备的一种复凝胶剂。渭北野生羊角蔓为萝呮科萝呮属萝呮的一个变种,渭北民间有人用其白汁治疗白癜风,《广济方》有同种植物治疗白癜风的记载。羊角蔓资源十分丰富,多年来一直是自生自灭,为了开发利用这一自然资源寻求治疗顽疾白癜风新药,本文对其进行了有效部位的筛选、成分检定、制备工艺、剂型制备及初步评价四方面的研究。本研究首先采用化学方法对羊角蔓所含化学成分进行系统预测,依据检测结果,用植物化学的研究方法得出单体成分并进行结构检定;其次综合研究结果,按极性分为4大类,用4种极性不同的溶剂进行分类提取,提取物采用活性导向分离法,筛选出有效部位;再次采用4因子3水平响应面法对有效部位的浸出工艺进行优化研究;最后采用试验对比、综合评价的方法进行制剂工艺研究,并用生物活性法进行药效学初步评价。主要结果如下:(1)羊角蔓治疗白癜风有效部位筛选通过系统预测,定性鉴别得出羊角蔓中含有皂苷、三萜类、甾体类、甾醇、黄酮类、生物碱和有机酸等14类化学成分。通过验证建立的酪氨酸酶活性测定法对石油醚提取物、乙酸乙酯提取物、正丁醇提取物、水提20%、40%、60%、80%、95%醇沉物8类不同极性组合物进行生化测定,得出在试验范围内,羊角蔓正丁醇提取物对酪氨酸酶激活效果最为显著,其激活率为30.71%,阳性对照补骨脂素激活率为39.34%。(2)羊角蔓有效部位的植化研究正丁醇提取物,经理化检测,富含皂苷,薄层色谱检测含2个皂苷斑点;经柱色谱分离、结晶纯化等处理,得到4个化合物单体,经理化、色谱、光谱等方法检定,得出化合物1为胡萝卜苷,化合物2为β-谷甾醇,化合物3为刺槐苷(山奈酚-3-O-葡萄糖基-鼠李糖基-7-O-鼠李糖苷),化合物4为肌醇。(3)羊角蔓有效部位的浸出研究经显微观察得出羊角蔓藤茎组织结构主要为表皮薄壁细胞、皮层薄壁细胞、次生韧皮部薄壁细胞、次生木质部薄壁细胞及髓细胞;叶片组织结构主要为表皮薄壁细胞和叶肉细胞。组织化学定位得出羊角蔓藤茎中皂苷主要分布在其皮层薄壁细胞、次生韧皮部薄壁细胞和髓细胞中;叶片中皂苷主要分布在叶肉细胞中。浸出工艺研究,在单因素研究的基础上,以总皂苷的标示量为效应指标,通过响应面中心设计试验进行工艺优化,得出在试验范围内的最佳提取工艺条件为:乙醇浓度76%,液料比10.32:1(mL/g),回流温度70.2℃,回流时间2.52 h;5次试验,固形物收率为4.32%,产物中总皂苷的标示量为60.39%。经验证,该工艺的相对标准偏差RSD为1.50%。(4)制剂研究及药效初步评价以正丁醇提取物的理化性质为依据,以临床适用性为判定标准,经文献调研和6种外用剂型对比分析得出w/o/w复乳凝胶剂对本产品较为适合。制剂处方和成型工艺参数经L9(34)正交试验优化得出,以1.0 g药物,加1.2 g单硬脂酸甘油酯,0.5 g白凡士林,2.7 g液体石蜡,0.25 g硬脂酸,0.2 g司盘-80,0.1 g吐温-80,0.1 g CMC-Na,0.1 g PVA124及少量助剂和适量蒸馏水,80℃乳化,制成的产品呈米黄色,稳定性良好,功效成分以刺槐苷计,其标示量不低于8.0%,释药性为0~8 h内,药物累积释放率达75.6%,体外释药方式中一级模型的相关系数较高。药效初步评价试验结果表明,该产品对皮肤无刺激;对酪氨酸酶的激活率为30.62%,阳性对照复方补骨脂酊激活率为57.34%。由此得出结论:渭北野生羊角蔓治疗白癜风有效部位为苷类物质,以总皂苷为指标,标示量不低于60%;羊角蔓中含有胡萝卜苷、β-谷甾醇、刺槐苷、肌醇和2个皂苷类化合物,其中肌醇为在同种植物中初次发现;乙醇提取正丁醇萃取工艺稳定,可操作性强;复凝胶剂对酪氨酸酶激活作用可靠;本研究的结果为开发治疗白癜风新药提供了可靠的试验依据。
[Abstract]:Leucoconjugation is a kind of compound gel which is prepared from the effective part of the wild Ramulus Ramulus in Weibei. Wild Ramulus Ramulus Ramulus in Weibei is a variety of Ramulus Ramulus. Some people in Weibei have used its white juice to treat vitiligo. There are records of treating vitiligo with the same plant. In order to exploit and utilize this natural resource to seek a new medicine for vitiligo, this paper studied its effective parts screening, ingredient identification, preparation process, preparation and preliminary evaluation of four aspects. According to the test results, the monomer components were obtained by phytochemistry and their structures were examined; secondly, according to the polarity, the monomer components were divided into four categories and extracted by four different polar solvents, and the active fractions were screened out by activity-oriented separation method; and then the effective fractions were extracted by 4-factor-3 horizontal response surface methodology. The main results are as follows: (1) Screening the effective parts of Vitiligo Vitiligo treatment by systematic prediction, qualitative identification of Vitiligo Vitiligo contains saponins, triterpenoids, steroids, steroids, steroids, steroids. Tyrosinase activity assay was established to determine petroleum ether extract, ethyl acetate extract, n-butanol extract, water extract 20%, 40%, 60%, 80%, 95% alcohol sediment and eight kinds of different polar compositions. The activation rate of tyrosinase was 30.71%. The activation rate of Psoralen in positive control was 39.34%. (2) Phytochemical study on the effective fraction of Aristolochia tenuifolia L. Physicochemical, chromatographic and spectroscopic analysis showed that compound 1 was carotene, compound 2 was beta-sitosterol, compound 3 was Acacia glycoside (kaempferol-3-O-glucosyl-rhamnosyl-7-O-rhamnosyl glycoside), compound 4 was inositol. (3) Microscopic observation of the effective parts of Ramulus angustifolia stem tissue structure was mainly surface. Histochemical localization showed that saponins were mainly distributed in cortical parenchyma cells, secondary phloem parenchyma cells, secondary xylem parenchyma cells and pulp cells. Saponins were mainly distributed in mesophyll cells. On the basis of single factor study, the extraction process was optimized by response surface methodology. The optimum extraction conditions were as follows: ethanol concentration 76%, liquid-to-material ratio 10.32:1 (mL/g), reflux temperature 70.2 C and reflux temperature 70.2 C. The relative standard deviation (RSD) was 1.50%. (4) The preparation study and pharmacodynamics preliminary evaluation were based on the physicochemical properties of n-butanol extract, and the clinical applicability was taken as the criterion. Literature investigation and comparison of six external dosage forms were carried out. The formulation and molding parameters were optimized by L9 (34) orthogonal test. 1.0 g drug, 1.2 g glycerin monostearate, 0.5 g white vaseline, 2.7 g liquid paraffin, 0.25 g stearic acid, 0.2 g spark-80, 0.1 g Tween-80, 0.1 g CMC-Na, 0.1 g PVA124 and a small amount of additives and suitable additives were obtained. The results of preliminary pharmacodynamic evaluation showed that the product had no irritation to the skin, and the cumulative release rate was 75.6% within 0-8 hours. The activation rate of tyrosinase was 30.62%. The activation rate of positive control compound psoralen tincture was 57.34%. It was concluded that the effective parts of wild Ramulus amurensis in Weibei were glycosides, and the total saponins were used as indicators, the labeling quantity was not less than 60%. Ramulus amurensis contained carotene, beta-sitosterol, Acacia glycosides, inositol and two saponins. Mesoinositol was found in the same plant for the first time; ethanol extraction of n-butanol extraction process is stable and operable; compound gel can activate tyrosinase reliably; the results of this study provide a reliable experimental basis for the development of new drugs for vitiligo.
【学位授予单位】：陕西科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TQ460.6

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