黄芩有效部位鼻用脑靶向制剂研究
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摘要
本课题以黄芩有效部位为原料,基于“鼻脑通路”的理论和实践,以“安全、有效、稳定、可控”为指导原则,将其研制成鼻用黄芩有效部位磷脂复合物亚微乳制剂,通过鼻腔给药用于脑缺血损伤的防治。通过对黄芩有效部位的提取纯化工艺、有效部位筛选评价、有效部位的改性、给药途径和剂型筛选、成型工艺、质量控制、安全性有效性评价、体内动力学及吸收机制等内容的系统研究,为开发有效防治脑缺血损伤的中药新制剂奠定基础；探索了中药制剂通过鼻腔递药入脑的可行性。
首先,采用水提酸沉法制备得到不同黄芩苷含量的有效部位,采用体外抗氧化实验和药效学实验筛选黄芩的最佳有效部位,通过药效学实验确定了给药途径和有效剂量。确定的提取纯化工艺为：黄芩药材加水(沸水投料)提取3次,合并提取液,浓缩至0.8g/ml,加盐酸调节pH值至1.0～2.0,70℃保温1h,静置,取沉淀物加适量水分散,用20%氢氧化钠调节pH7,加入规定量乙醇(含醇量10%和40%),静置,滤过,滤液用盐酸调节pH1-2,80℃保温1小时,滤过,沉淀用相应乙醇洗涤至pH5,减压干燥,得到黄芩苷含量为为62%和85%的提取物,同时购买黄芩苷含量为98%的提取物。进行了上述三个规格提取物的体外抗氧化实验研究,并以线栓法复制的大鼠局灶性脑缺血模型优选三个规格的提取物,结果85%提取物具有较好的体外抗氧化表现和药效表现；同时对给药途径和有效剂量进行了研究,确定了鼻腔给药方式作为给药途径,初步确定了有效剂量。
鉴于黄芩有效部位水溶性、脂溶性均较差,难于透过生物膜而影响其生物利用度,采用磷脂复合物技术对有效部位进行了改性处理,结果显著改善了黄芩有效部位的油水分配系数从而改善生物利用度。通过星点设计-效应面优化的磷脂复合物制备工艺：以无水乙醇为复合溶剂,磷脂与黄芩有效部位投料比为2：1,黄芩有效部位浓度为3.0mg/ml,反应温度为50℃。
在剂型筛选研究中,结合磷脂复合物的性质和鼻腔给药的特点,通过猪鼻黏膜透过性能与黏膜刺激性实验对油溶液剂、水溶液剂、微乳、亚微乳和原位凝胶5种剂型进行筛选研究。在猪鼻黏膜透过试验中,亚微乳表现出良好的透过能力,其表观渗透系数为(37.09±1.75)×10-3(μg·cm-2.s-1),明显优于其它剂型；蟾蜍上腭纤毛刺激性和大鼠鼻黏膜刺激性实验结果表明,亚微乳是较为理想鼻腔给药剂型。
采用正交设计及星点设计-效应面优化等实验设计方法,对黄芩有效部位磷脂复合物亚微乳的处方因素和制备工艺进行研究,确定了亚微乳的制剂处方和工艺：取黄芩有效部位与大豆卵磷脂置于反应釜中,加入无水乙醇,60℃水浴回流2h,充分复合。于60℃减压回收无水乙醇至剩余约30%,停止回收,加入中涟油、大豆油、乳化磷脂、油酸、维生素E等物料,继续减压回收溶剂至无醇味,得油相；将甘油、泊洛沙姆、乙二胺四乙酸钠等水相成分溶于适量预热的蒸馏水中,得水相；将水相徐徐加入油相中,充分混合,60。C的条件下以19000转/min的剪切速度剪切9min；冷却至室温后,于高压均质机上进行高压均质处理,均质压力为600bar,次数为6次,低压均质在60bar压力下均质3次,收集乳液,用0.1mol/L的氢氧化钠溶液调节pH5,微孔滤膜滤过即得亚微乳制剂。对制剂进行了影响因素试验,结果表明,该制剂适宜贮存条件为低温(4～8℃)避光保存。在制备工艺研究过程中,由于磷脂复合物不易干燥,且极易吸湿,同时复合物在油相中溶解度较小,投药量不能完全溶解和均匀分散在油相中,最终利用了专利技术溶质相转移技术成功的解决了溶解和分散的问题,制备得到高载药量的亚微乳,同时免去了磷脂复合物干燥工序,对转相技术进行了机制探讨。
按照中国药典2010年版相关要求,对制剂制定了严格的质量标准,进行了定性鉴别、检查和含量测定等项目进行了研究,建立了科学、可行的质控方法；对药物在制剂中的相分布、制剂的在体鼻腔吸收特征等进行了研究。结果提示,药物主要定位于磷脂膜和包裹于油相中,制剂的包封率达到了89.49%；制剂的大鼠在体鼻腔吸收符合一级动力学吸收特征,制剂120min的在体吸收量较磷脂复合物溶液吸收增加20%,吸收速度常数增加13%,表明制剂能够进一步增强黄芩有效部位磷脂复合物的吸收,属被动吸收机制。
采用双侧颈动脉阻断法建立大鼠脑缺血模型和血管内栓线阻断法建立大鼠局灶性脑缺血模型,评价了亚微乳对大鼠脑缺血损伤所致神经运动性损伤、脑梗死体积、脑水肿、病理组织学变化,脑组织与血液中的相关生化指标的影响。结果表明,制剂组与模型组比较,神经功能与脑组织病理学变化评分有显著差异(P<0.01),脑组织含水量减低(P<0.01),脑梗死体积比有一定程度改善；制剂能显著提高全双侧颈总动脉结扎脑缺血大鼠体内SOD、ATP酶水平,降低其MDA含量(P<0.01),对于GSH-PX、Ca2+-ATP酶、Na+-K+-ATP酶、LD和LDH酶水平没有显著升高高或降低,但是却有升高或降低的趋势；能显著提高局灶性脑缺血损伤大鼠体内SOD、Ca2+-ATP酶水平(P<0.05),对于GSH-PX酶和Na+-K+-ATP酶水平没有显著升高,但是却有升高的趋势。提示本制剂可能通过增强大鼠抗自由基能力,改善脑组织能量代谢来发挥对大鼠缺血性脑损伤的修复保护作用；研究了制剂的急性毒性和对大鼠鼻黏膜、蟾蜍上颚的刺激性,结果表明,未显示明显的刺激性和毒性反应。
以黄芩苷为评价指标,对亚微乳的脑内动力学及脑内转运路径进行了探索,结果表明,制剂通过鼻腔给药的绝对生物利用度达到了72%；制剂鼻腔给药脑中生物利用度是静脉给药的5.68倍,以静脉给药为参照,制剂的脑靶向指数为7.88,提示本制剂经鼻给药后,可显著提高黄芩苷在脑组织中的分布,具有明显的脑靶向性；通过对嗅球、大脑、小脑中药物浓度的变化和生物利用度的结果分析提示,黄芩苷主要是通过嗅神经通路传递入脑,即药物分子经过嗅黏膜吸收,被神经元轴突末梢摄取,经轴浆转运至嗅球,进一步达到嗅脑,再进入小脑、大脑等部位。
本课题以黄芩有效部位为研究对象,探索了中药有效成分鼻腔给药治疗脑内疾病的可行性,实践了难溶性中药有效成分的磷脂复合物改性技术,创新性的发明了高载药量亚微乳制剂的成型专利技术(转相技术),为中药难溶性成分制剂开发研究提供了借鉴,同时,为鼻腔给药途径防治脑部疾病的研究提供了良好的依据和参考。
We prepared a sub-microemulsion preparation of scutellaria effective fraction phospholipid complex in nasal systems which depended on the theory and pratice of "nasal-brain pathway" and effective parts of scutellaria as raw materials, and safe, stable, effective and controllbale as guiding principle for preventing and treating cerebral ischemic injury through nasal delivery. In order to lay the foundation for the traditional Chinese medicine preparation which prevent and treat erebral ischemic injury and investigate the feasibility of Chinese medicine preparation which deliver drug through nasal into brain, we took a system study obout the process of extraction and purification, preparation, quality control, the evaluation of safety and effectiveness, and absorption mechanism, and so on.
Firstly, we prepared effective parts that dfferent contents of baicalin from scutellaria by using water extraction and acid precipitation method, and screening specification of optimum effective parts by using antioxidation in vitro and pharmacodynamics test, and determining the route of administration and effective dose. The preparation technologies for effective parts are as the follows:scutellaria was decocted for three times in water (after boiling), combined extracts and concentrated0.8g/ml, added HCl to adjust to pHl.0-2.0, stood for1h under70℃and stirred precipitate which added proper water, adjusted its pH to7.0by20%NaOH, added specified amount of ethanol (the content of alcohol were10%and40%), stood, filtrated, adjusted filtrate's pH to1.0-2.0by HCl, stood for1h under80℃and cooled to room temperature, filtrated,washed precipitate's pH to5.0by ethanol and dryed by decompression drying to get sample which purity of62%and85%, in the same time, bought98%scutellaria extracts as. We researched different content of extracts which antioxidation in vitro and pharmacological test by intravascular suture blocking method to establish a focal cerebral ischemia., results showed85%extracts had the better effect of antioxidation and efficacy; explored the route of administration and effective dose, determined the better route was nasal administration and initially to determinn the effective dose.
In view of the water and lipid-soluble of scutellaria extracts are all poor and difficult through the biofilm to affect the bioavailability, we studied the modification of extracts by the technology of phosphlipid complex, results showed that significantly improved oil-water partition coefficient and bioavailability of extracts in phosplipid complex. The preparation technologies for phosplipid complex are as the follows:used absolute ethylalcohol as complex solvent, the ratio of phosplipid and extract was2:1, the extract concentration was3.0mg/ml, the reaction temperature was50℃.
Combined the propertise of phospholipid complex and characteristics of intranasal administration, we studied the screening of formulation by investigating swine nasal mucosa permeability and animal nasal mucosa irritation for oil solution, aqueous solution, microemulsion, sub-microemulsion and insitu gel. Sub-microemulsion had the better permeability in the test of swine nasal mucosa permeability, the apparent permeability coefficient is37.09±1.75×10-3(μg·cm-2·s-1), significantly greater than other formulations; investigated the toad palate mucomembranous ciliotoxicity of formulation and its stimulation on the rat nasal mucosa, results showed sub-microemulsion is the best form for scutellaria extract phosplipid complex in nasal systems.
We studied the formulation factors and preparation processes of scutellaria extract phosplipid complex sub-microemulsion, and determined the prescription of preparation by using orthogonal design method and central composite design-response surface optimization. The preparation technologies are as the follows:weighed scutellaria extracts, sobean lecithin as above the prescription, placed at a reactor, added to absolute ethanol, refluxed in a kettle at60℃that reaction time was2h, fully composite, recovered solvent by vacuum concentration to get the remining amount of ethanol was about30%, then added the medium-chain triglycerides, soybean oil, emulsified phosphoilpids, oleic acid, vitamine E, and continued recovered to without ethanol taste, which was the oil phase. Glycerin, poloxamer, sodium EDTA were dissolved in appropiate amount of warm-up distilled water, which was the aqueous phase. Two-phase was heated to60℃respectively, added the aqueous phase to the oil phase, then mixed. Colostrum was sheared for9min by19000rev/min at60℃, and cooled to room temperature, conducted homogenization in high pressure homogenizer for6times that high pressure homogenization pressure was600bar, for3times that low pressure homogenization pressure was60bar, adjusted its pH to5.0by0.1mol/l NaOH, and filtrated, which was scutellaria extract phosplipid complex sub-microemulsion. Influence factors test results showed the appropriate storage condition of the formulation is low temperature(4-8℃) keeping away from direct sunlight; In the research of preparation,because phospholipids complex is hard to dry and easy to absorb water, at the same time, complex in oil phase solubility smaller, addition mount of durg can't completely dissolved and uniform dispersion in oil phase, finally, we used the patent technology solute phase transfer technology successfully solved the problem of dissolved and scattered, got the high drug loading of the submic ro emulsion, meanwhile, avoid the phospholipids complex dry process, and explored t he mechanism of phase transfer technology.
According to the pharmacopoeia of the People's Republic of China on the2010edition, we made further study about the quality standards of sub-microemulsion, including qualitative identification, general quality inspection, and content determination. At the end we established scientific and feasible quality control methods. Results that physical and chemical properties research showed preparation process was rational and feasible that encapsulation efficiency of preparation was89.49%; nasal absorption accorded with first order kinetic and belonged to a passive absorption.
We researched the effcet of scutellaria extract phosplipid complex sub-microemulsion on cerebral ischemia in rats by used bilateral carotid artery blocking method to establish a complete cerebral ischemia and intravascular suture blocking method to establish a focal cerebral ischemia. Results showed that compared with the model group, Brain indexes of the drug-treated group were all lower than that of the model group (P<0.01), the content of brain water was significantly decreased (P<0.01, P<0.05), brain tissue pathomorphology and nerve function were markedly improved (P<0.01), neurological deficits scores were significantly decreased(P<0.01); the content of SOD and ATPase were increased, while the content of MDA was significantly deceased in drug-treated group (P<0.01) for complete cerebral ischemia in rats; the content of SOD and Ca2+-ATP were highly increased (P <0.05) for focal cerebral ischemia in rats. Results of toxicity and irritation tests showed that sub-microemulsion has no obvious toxicity.
With baicalin as a evaluation index, we studied the brain dynamics and absorption mechanism of sub-microemulsion. Results showed that plasma bioavailability of the nasal adminstration was72%; brain targeting index was7.88which comparaed with intravenous administration; the main way into the brain was olfactory pathway that drug molecules were absorbed through the olfactory mucosa, the uptake of which by neuron axon terminals, arrived at olfactory bulb through axoplasm, further to reach the olfactory, then enter the cerebellum, the brain and other parts.
We used effective parts of scutellaria as research object to explore the feasibility of traditional Chinese medicine by intranasal administration for treat diseases of the brain, practice insoluble active ingredients of traditional Chinese medicine phospholipid complex modifid technology, innnovatively invented the formulations molding patented technology(phase conversion technique) that high drug loading sub-microemulsion. This paper provides a basis and reference for the development of traditional Chinese medicine insoluble components and the prevention and treatment for cerebrovascular diseases by nasal drug delivery.
首先,采用水提酸沉法制备得到不同黄芩苷含量的有效部位,采用体外抗氧化实验和药效学实验筛选黄芩的最佳有效部位,通过药效学实验确定了给药途径和有效剂量。确定的提取纯化工艺为：黄芩药材加水(沸水投料)提取3次,合并提取液,浓缩至0.8g/ml,加盐酸调节pH值至1.0～2.0,70℃保温1h,静置,取沉淀物加适量水分散,用20%氢氧化钠调节pH7,加入规定量乙醇(含醇量10%和40%),静置,滤过,滤液用盐酸调节pH1-2,80℃保温1小时,滤过,沉淀用相应乙醇洗涤至pH5,减压干燥,得到黄芩苷含量为为62%和85%的提取物,同时购买黄芩苷含量为98%的提取物。进行了上述三个规格提取物的体外抗氧化实验研究,并以线栓法复制的大鼠局灶性脑缺血模型优选三个规格的提取物,结果85%提取物具有较好的体外抗氧化表现和药效表现；同时对给药途径和有效剂量进行了研究,确定了鼻腔给药方式作为给药途径,初步确定了有效剂量。
鉴于黄芩有效部位水溶性、脂溶性均较差,难于透过生物膜而影响其生物利用度,采用磷脂复合物技术对有效部位进行了改性处理,结果显著改善了黄芩有效部位的油水分配系数从而改善生物利用度。通过星点设计-效应面优化的磷脂复合物制备工艺：以无水乙醇为复合溶剂,磷脂与黄芩有效部位投料比为2：1,黄芩有效部位浓度为3.0mg/ml,反应温度为50℃。
在剂型筛选研究中,结合磷脂复合物的性质和鼻腔给药的特点,通过猪鼻黏膜透过性能与黏膜刺激性实验对油溶液剂、水溶液剂、微乳、亚微乳和原位凝胶5种剂型进行筛选研究。在猪鼻黏膜透过试验中,亚微乳表现出良好的透过能力,其表观渗透系数为(37.09±1.75)×10-3(μg·cm-2.s-1),明显优于其它剂型；蟾蜍上腭纤毛刺激性和大鼠鼻黏膜刺激性实验结果表明,亚微乳是较为理想鼻腔给药剂型。
采用正交设计及星点设计-效应面优化等实验设计方法,对黄芩有效部位磷脂复合物亚微乳的处方因素和制备工艺进行研究,确定了亚微乳的制剂处方和工艺：取黄芩有效部位与大豆卵磷脂置于反应釜中,加入无水乙醇,60℃水浴回流2h,充分复合。于60℃减压回收无水乙醇至剩余约30%,停止回收,加入中涟油、大豆油、乳化磷脂、油酸、维生素E等物料,继续减压回收溶剂至无醇味,得油相；将甘油、泊洛沙姆、乙二胺四乙酸钠等水相成分溶于适量预热的蒸馏水中,得水相；将水相徐徐加入油相中,充分混合,60。C的条件下以19000转/min的剪切速度剪切9min；冷却至室温后,于高压均质机上进行高压均质处理,均质压力为600bar,次数为6次,低压均质在60bar压力下均质3次,收集乳液,用0.1mol/L的氢氧化钠溶液调节pH5,微孔滤膜滤过即得亚微乳制剂。对制剂进行了影响因素试验,结果表明,该制剂适宜贮存条件为低温(4～8℃)避光保存。在制备工艺研究过程中,由于磷脂复合物不易干燥,且极易吸湿,同时复合物在油相中溶解度较小,投药量不能完全溶解和均匀分散在油相中,最终利用了专利技术溶质相转移技术成功的解决了溶解和分散的问题,制备得到高载药量的亚微乳,同时免去了磷脂复合物干燥工序,对转相技术进行了机制探讨。
按照中国药典2010年版相关要求,对制剂制定了严格的质量标准,进行了定性鉴别、检查和含量测定等项目进行了研究,建立了科学、可行的质控方法；对药物在制剂中的相分布、制剂的在体鼻腔吸收特征等进行了研究。结果提示,药物主要定位于磷脂膜和包裹于油相中,制剂的包封率达到了89.49%；制剂的大鼠在体鼻腔吸收符合一级动力学吸收特征,制剂120min的在体吸收量较磷脂复合物溶液吸收增加20%,吸收速度常数增加13%,表明制剂能够进一步增强黄芩有效部位磷脂复合物的吸收,属被动吸收机制。
采用双侧颈动脉阻断法建立大鼠脑缺血模型和血管内栓线阻断法建立大鼠局灶性脑缺血模型,评价了亚微乳对大鼠脑缺血损伤所致神经运动性损伤、脑梗死体积、脑水肿、病理组织学变化,脑组织与血液中的相关生化指标的影响。结果表明,制剂组与模型组比较,神经功能与脑组织病理学变化评分有显著差异(P<0.01),脑组织含水量减低(P<0.01),脑梗死体积比有一定程度改善；制剂能显著提高全双侧颈总动脉结扎脑缺血大鼠体内SOD、ATP酶水平,降低其MDA含量(P<0.01),对于GSH-PX、Ca2+-ATP酶、Na+-K+-ATP酶、LD和LDH酶水平没有显著升高高或降低,但是却有升高或降低的趋势；能显著提高局灶性脑缺血损伤大鼠体内SOD、Ca2+-ATP酶水平(P<0.05),对于GSH-PX酶和Na+-K+-ATP酶水平没有显著升高,但是却有升高的趋势。提示本制剂可能通过增强大鼠抗自由基能力,改善脑组织能量代谢来发挥对大鼠缺血性脑损伤的修复保护作用；研究了制剂的急性毒性和对大鼠鼻黏膜、蟾蜍上颚的刺激性,结果表明,未显示明显的刺激性和毒性反应。
以黄芩苷为评价指标,对亚微乳的脑内动力学及脑内转运路径进行了探索,结果表明,制剂通过鼻腔给药的绝对生物利用度达到了72%；制剂鼻腔给药脑中生物利用度是静脉给药的5.68倍,以静脉给药为参照,制剂的脑靶向指数为7.88,提示本制剂经鼻给药后,可显著提高黄芩苷在脑组织中的分布,具有明显的脑靶向性；通过对嗅球、大脑、小脑中药物浓度的变化和生物利用度的结果分析提示,黄芩苷主要是通过嗅神经通路传递入脑,即药物分子经过嗅黏膜吸收,被神经元轴突末梢摄取,经轴浆转运至嗅球,进一步达到嗅脑,再进入小脑、大脑等部位。
本课题以黄芩有效部位为研究对象,探索了中药有效成分鼻腔给药治疗脑内疾病的可行性,实践了难溶性中药有效成分的磷脂复合物改性技术,创新性的发明了高载药量亚微乳制剂的成型专利技术(转相技术),为中药难溶性成分制剂开发研究提供了借鉴,同时,为鼻腔给药途径防治脑部疾病的研究提供了良好的依据和参考。
We prepared a sub-microemulsion preparation of scutellaria effective fraction phospholipid complex in nasal systems which depended on the theory and pratice of "nasal-brain pathway" and effective parts of scutellaria as raw materials, and safe, stable, effective and controllbale as guiding principle for preventing and treating cerebral ischemic injury through nasal delivery. In order to lay the foundation for the traditional Chinese medicine preparation which prevent and treat erebral ischemic injury and investigate the feasibility of Chinese medicine preparation which deliver drug through nasal into brain, we took a system study obout the process of extraction and purification, preparation, quality control, the evaluation of safety and effectiveness, and absorption mechanism, and so on.
Firstly, we prepared effective parts that dfferent contents of baicalin from scutellaria by using water extraction and acid precipitation method, and screening specification of optimum effective parts by using antioxidation in vitro and pharmacodynamics test, and determining the route of administration and effective dose. The preparation technologies for effective parts are as the follows:scutellaria was decocted for three times in water (after boiling), combined extracts and concentrated0.8g/ml, added HCl to adjust to pHl.0-2.0, stood for1h under70℃and stirred precipitate which added proper water, adjusted its pH to7.0by20%NaOH, added specified amount of ethanol (the content of alcohol were10%and40%), stood, filtrated, adjusted filtrate's pH to1.0-2.0by HCl, stood for1h under80℃and cooled to room temperature, filtrated,washed precipitate's pH to5.0by ethanol and dryed by decompression drying to get sample which purity of62%and85%, in the same time, bought98%scutellaria extracts as. We researched different content of extracts which antioxidation in vitro and pharmacological test by intravascular suture blocking method to establish a focal cerebral ischemia., results showed85%extracts had the better effect of antioxidation and efficacy; explored the route of administration and effective dose, determined the better route was nasal administration and initially to determinn the effective dose.
In view of the water and lipid-soluble of scutellaria extracts are all poor and difficult through the biofilm to affect the bioavailability, we studied the modification of extracts by the technology of phosphlipid complex, results showed that significantly improved oil-water partition coefficient and bioavailability of extracts in phosplipid complex. The preparation technologies for phosplipid complex are as the follows:used absolute ethylalcohol as complex solvent, the ratio of phosplipid and extract was2:1, the extract concentration was3.0mg/ml, the reaction temperature was50℃.
Combined the propertise of phospholipid complex and characteristics of intranasal administration, we studied the screening of formulation by investigating swine nasal mucosa permeability and animal nasal mucosa irritation for oil solution, aqueous solution, microemulsion, sub-microemulsion and insitu gel. Sub-microemulsion had the better permeability in the test of swine nasal mucosa permeability, the apparent permeability coefficient is37.09±1.75×10-3(μg·cm-2·s-1), significantly greater than other formulations; investigated the toad palate mucomembranous ciliotoxicity of formulation and its stimulation on the rat nasal mucosa, results showed sub-microemulsion is the best form for scutellaria extract phosplipid complex in nasal systems.
We studied the formulation factors and preparation processes of scutellaria extract phosplipid complex sub-microemulsion, and determined the prescription of preparation by using orthogonal design method and central composite design-response surface optimization. The preparation technologies are as the follows:weighed scutellaria extracts, sobean lecithin as above the prescription, placed at a reactor, added to absolute ethanol, refluxed in a kettle at60℃that reaction time was2h, fully composite, recovered solvent by vacuum concentration to get the remining amount of ethanol was about30%, then added the medium-chain triglycerides, soybean oil, emulsified phosphoilpids, oleic acid, vitamine E, and continued recovered to without ethanol taste, which was the oil phase. Glycerin, poloxamer, sodium EDTA were dissolved in appropiate amount of warm-up distilled water, which was the aqueous phase. Two-phase was heated to60℃respectively, added the aqueous phase to the oil phase, then mixed. Colostrum was sheared for9min by19000rev/min at60℃, and cooled to room temperature, conducted homogenization in high pressure homogenizer for6times that high pressure homogenization pressure was600bar, for3times that low pressure homogenization pressure was60bar, adjusted its pH to5.0by0.1mol/l NaOH, and filtrated, which was scutellaria extract phosplipid complex sub-microemulsion. Influence factors test results showed the appropriate storage condition of the formulation is low temperature(4-8℃) keeping away from direct sunlight; In the research of preparation,because phospholipids complex is hard to dry and easy to absorb water, at the same time, complex in oil phase solubility smaller, addition mount of durg can't completely dissolved and uniform dispersion in oil phase, finally, we used the patent technology solute phase transfer technology successfully solved the problem of dissolved and scattered, got the high drug loading of the submic ro emulsion, meanwhile, avoid the phospholipids complex dry process, and explored t he mechanism of phase transfer technology.
According to the pharmacopoeia of the People's Republic of China on the2010edition, we made further study about the quality standards of sub-microemulsion, including qualitative identification, general quality inspection, and content determination. At the end we established scientific and feasible quality control methods. Results that physical and chemical properties research showed preparation process was rational and feasible that encapsulation efficiency of preparation was89.49%; nasal absorption accorded with first order kinetic and belonged to a passive absorption.
We researched the effcet of scutellaria extract phosplipid complex sub-microemulsion on cerebral ischemia in rats by used bilateral carotid artery blocking method to establish a complete cerebral ischemia and intravascular suture blocking method to establish a focal cerebral ischemia. Results showed that compared with the model group, Brain indexes of the drug-treated group were all lower than that of the model group (P<0.01), the content of brain water was significantly decreased (P<0.01, P<0.05), brain tissue pathomorphology and nerve function were markedly improved (P<0.01), neurological deficits scores were significantly decreased(P<0.01); the content of SOD and ATPase were increased, while the content of MDA was significantly deceased in drug-treated group (P<0.01) for complete cerebral ischemia in rats; the content of SOD and Ca2+-ATP were highly increased (P <0.05) for focal cerebral ischemia in rats. Results of toxicity and irritation tests showed that sub-microemulsion has no obvious toxicity.
With baicalin as a evaluation index, we studied the brain dynamics and absorption mechanism of sub-microemulsion. Results showed that plasma bioavailability of the nasal adminstration was72%; brain targeting index was7.88which comparaed with intravenous administration; the main way into the brain was olfactory pathway that drug molecules were absorbed through the olfactory mucosa, the uptake of which by neuron axon terminals, arrived at olfactory bulb through axoplasm, further to reach the olfactory, then enter the cerebellum, the brain and other parts.
We used effective parts of scutellaria as research object to explore the feasibility of traditional Chinese medicine by intranasal administration for treat diseases of the brain, practice insoluble active ingredients of traditional Chinese medicine phospholipid complex modifid technology, innnovatively invented the formulations molding patented technology(phase conversion technique) that high drug loading sub-microemulsion. This paper provides a basis and reference for the development of traditional Chinese medicine insoluble components and the prevention and treatment for cerebrovascular diseases by nasal drug delivery.
引文
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