阿昔洛韦脂质体滴眼剂的研究
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摘要
传统滴眼剂的主要缺点是生物利用度低，入眼后很快被泪液清除。相比之下，脂质体能够延长药物在眼部的滞留时间，增强药物的角膜穿透能力，从而大大提高药物的生物利用度。本文以阿昔洛韦为模型药物，将其制成脂质体滴眼剂，并进行了体内外的相关研究。
采用氯仿萃取分离药物与膜材，通过紫外分光光度法测定了脂质体中总药物含量，采用鱼精蛋白沉淀-紫外分光光度法测定游离药量。方法操作简单、测定准确、重现性好，符合分析要求。
比较了乙醇注入法、薄膜分散法、反相蒸发法、pH梯度法和中和法等不同的脂质体制备方法，结果发现这些方法均未能极大提高模型药物在脂质体中的包封率。经权衡比较后，最后选用包封率较高、稳定性较好的反相蒸发法制备阿昔洛韦脂质体。在单因素考察的基础上，本文进行了均匀设计实验，以包封率为指标，兼顾稳定性，筛选出最佳处方，包封率可达62％。加速实验表明，该制剂在低温、充氮、避光条件下贮存，稳定性较好。
本文考察了脂质体的理化性质，如粒径分布、表面电位、黏度、表面张力等。测得其平均粒径为151.9nm(number mean)。流变学性质属于非牛顿流体，并且随温度的升高黏度下降。表面张力为57.58×10~(-3)N／m。表面电位为-78.87mV。
运用γ-闪烁照相技术研究脂质体在角膜的在体滞留能力，结果表明，同空白缓冲液相比，脂质体组能够明显延长角膜滞留时间。滞留能力正电性脂质体>负电性脂质体>空白缓冲液，半衰期t_(1／2)分别为51.52min，14.75min和4.18min。
一
建立兔眼单纯疮疹病毒角膜炎模型，以兔眼角膜病变范围和病毒滴
度厂 O为指标，考察了制剂的药效。结果显示，脂质体制剂疗效明显
优于市售药物水溶液组。与空白对照组相比，有极显著差异叩4刀1人
可明显抑制病情的发展，降低病毒滴度，有效率达 100％。局部眼刺激性实
验证明，该制剂无刺激性。
The main drawback of traditional eye drops is their poor bioavailability. After administration, they are diminished by tear fluid quickly. Compared with them , liposomal eye drops can prolong the residence time in precorneal and build the ability to penetrate into aqueous humor therefore promote the bioavailbility .Using acyclovir as model drug,liposomal eye drops were designed and a series of relative researches were carried out.
A method was developed to determine the encapsulation efficiency of ACV-containing liposome preparations by chloroform extration-UV spectrophotometry and protamine precipitation-UV spectrophotometry. The method was simple and accurate and easy to operate.
The Reverse-Phase evaporation (REV) method was employed for the manufacture of acyclovir liposomes from natural soyabean lethicin. The Encapsulation Efficiency (EN) of acydovir-containing liposome (ACVL) was determined and some factors which may affect the EN such as the
drug-lipid ratio, ionic strength were investgated.
In order to ensure excellent stability and to obtain higher encapsulation efficiency, a uniform design was carried out and obtained the optimum component of liposome and the average encapsulation efficiency was above 62%. In accelerated tests, it was found that the liposome suspension was stable at low temperature in oxygen-free atmosphere and protected from light.
The physicochemical properties of liposomes were studied. The
morphological characteristic of liposome samples was revealed under transmission electron microscope. The surface electric potential , pH, and the surface tension of acyclovir liposomes is -78.87 mV,7.36, 57.58+10-3N/m respectively. The rheological curve proved that ACVL was non-Newtonian fluid and its viscosity was decreased by high temperature.
The clearance of eyedrop was found to exhibit a biphasic pattern by y-scintigraphic technique. At the end of 7 min only less than 20% of the applied solution was in contact with the corneal surface. For positive and negative liposomes,the residence time of radioactive marker was significantly prolonged and the retentive effect was superior to blank solution.The elimination half-life time of positive, negative and PBS was 51.52min, 14.75min and 4.18min respectively.
The toxicity of ACV liposome preparation was investigated by using index of eye irritation experiments .The conclusion was that the liposome preparation had no irritation.
The effects of ACV -containing liposome preparation on herpes simplex keratitis in conscious rabbits indicated that the liposome preparation markedly increased the index of treatment and was more effective than current acyclovir eye drops. Compared with the other two groups, there was statistical significance among them. Virus was isolated and TCID50 (tissue culture infective dose) was calculated. The results showed the same conclusion as clinical observance.
采用氯仿萃取分离药物与膜材，通过紫外分光光度法测定了脂质体中总药物含量，采用鱼精蛋白沉淀-紫外分光光度法测定游离药量。方法操作简单、测定准确、重现性好，符合分析要求。
比较了乙醇注入法、薄膜分散法、反相蒸发法、pH梯度法和中和法等不同的脂质体制备方法，结果发现这些方法均未能极大提高模型药物在脂质体中的包封率。经权衡比较后，最后选用包封率较高、稳定性较好的反相蒸发法制备阿昔洛韦脂质体。在单因素考察的基础上，本文进行了均匀设计实验，以包封率为指标，兼顾稳定性，筛选出最佳处方，包封率可达62％。加速实验表明，该制剂在低温、充氮、避光条件下贮存，稳定性较好。
本文考察了脂质体的理化性质，如粒径分布、表面电位、黏度、表面张力等。测得其平均粒径为151.9nm(number mean)。流变学性质属于非牛顿流体，并且随温度的升高黏度下降。表面张力为57.58×10~(-3)N／m。表面电位为-78.87mV。
运用γ-闪烁照相技术研究脂质体在角膜的在体滞留能力，结果表明，同空白缓冲液相比，脂质体组能够明显延长角膜滞留时间。滞留能力正电性脂质体>负电性脂质体>空白缓冲液，半衰期t_(1／2)分别为51.52min，14.75min和4.18min。
一
建立兔眼单纯疮疹病毒角膜炎模型，以兔眼角膜病变范围和病毒滴
度厂 O为指标，考察了制剂的药效。结果显示，脂质体制剂疗效明显
优于市售药物水溶液组。与空白对照组相比，有极显著差异叩4刀1人
可明显抑制病情的发展，降低病毒滴度，有效率达 100％。局部眼刺激性实
验证明，该制剂无刺激性。
The main drawback of traditional eye drops is their poor bioavailability. After administration, they are diminished by tear fluid quickly. Compared with them , liposomal eye drops can prolong the residence time in precorneal and build the ability to penetrate into aqueous humor therefore promote the bioavailbility .Using acyclovir as model drug,liposomal eye drops were designed and a series of relative researches were carried out.
A method was developed to determine the encapsulation efficiency of ACV-containing liposome preparations by chloroform extration-UV spectrophotometry and protamine precipitation-UV spectrophotometry. The method was simple and accurate and easy to operate.
The Reverse-Phase evaporation (REV) method was employed for the manufacture of acyclovir liposomes from natural soyabean lethicin. The Encapsulation Efficiency (EN) of acydovir-containing liposome (ACVL) was determined and some factors which may affect the EN such as the
drug-lipid ratio, ionic strength were investgated.
In order to ensure excellent stability and to obtain higher encapsulation efficiency, a uniform design was carried out and obtained the optimum component of liposome and the average encapsulation efficiency was above 62%. In accelerated tests, it was found that the liposome suspension was stable at low temperature in oxygen-free atmosphere and protected from light.
The physicochemical properties of liposomes were studied. The
morphological characteristic of liposome samples was revealed under transmission electron microscope. The surface electric potential , pH, and the surface tension of acyclovir liposomes is -78.87 mV,7.36, 57.58+10-3N/m respectively. The rheological curve proved that ACVL was non-Newtonian fluid and its viscosity was decreased by high temperature.
The clearance of eyedrop was found to exhibit a biphasic pattern by y-scintigraphic technique. At the end of 7 min only less than 20% of the applied solution was in contact with the corneal surface. For positive and negative liposomes,the residence time of radioactive marker was significantly prolonged and the retentive effect was superior to blank solution.The elimination half-life time of positive, negative and PBS was 51.52min, 14.75min and 4.18min respectively.
The toxicity of ACV liposome preparation was investigated by using index of eye irritation experiments .The conclusion was that the liposome preparation had no irritation.
The effects of ACV -containing liposome preparation on herpes simplex keratitis in conscious rabbits indicated that the liposome preparation markedly increased the index of treatment and was more effective than current acyclovir eye drops. Compared with the other two groups, there was statistical significance among them. Virus was isolated and TCID50 (tissue culture infective dose) was calculated. The results showed the same conclusion as clinical observance.
引文
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