8-甲氧基补骨脂素皮肤靶向纳米脂质体凝胶的研究及评价
|
摘要
白癜风是由于皮肤和毛囊的黑素细胞内酪氨酸系统的功能减退、丧失引起的,是一种以局限性或泛发性皮肤色素脱失为特征的疾病。本研究通过对8-甲氧基补骨脂素(MOP)皮肤靶向纳米脂质体凝胶制备工艺、靶向性、影响因素、质量评价标准、稳定性进行研究,旨在制备具有皮肤靶向性的MOP脂质体凝胶,进一步提高MOP局部外用治疗白癜风的效果,减少毒副作用,并探讨其靶向机理及影响因素,为开发高效、低毒、方便的白癜风治疗药物打下基础。
试验以包封率、平均粒径为主要考察指标,分别采用逆相蒸发法、干膜振荡分散法、薄膜-超声法、乙醚注入法制备MOP脂质体进行比较,确定薄膜-超声法为佳,并确定高压均质法制备纳米脂质体的工艺为1000bar均质6次。以卡波姆(CP)为基质,采用基质形成前加入脂质体的方法制备了脂质体凝胶,并对脂质体、脂质体凝胶的质量进行全面评价,认为脂质体及其凝胶制备工艺基本稳定、质量可控,用负染法、扫描电镜法对脂质体形态进行表征,脂质体颗粒圆整、均匀,并用冷冻蚀刻法对脂质体凝胶形态进行表征。激光散射法测定脂质体及脂质体凝胶的平均粒径分别为280nm、332nm,葡聚糖凝胶柱法测定5批供试样品包封率分别为94.2%、97.5%、93.4%、95.3%、95.7%,含量分别为2.96、2.82、2.88、3.03、3.11mg·mL~(-1)。2批脂质体凝胶的包封率分别为91.5%、87.7%,含量为1.82、1.93mg·g~(-1)。脂质体稳定性研究表明脂质体及其凝胶剂在4℃、25℃贮存90天稳定,对高温及强光不稳定。
体外透皮实验表明滞留于皮肤内的药量MOP脂质体比MOP搽剂高2.7倍,较脂质体凝胶高1.5倍,MOP脂质体凝胶比MOP搽剂高1.7倍;电镜观察表明脂质体不是直接透过皮肤进入血液,而是在皮肤中代谢、释放药物,然后药物再渗透、穿过皮肤。
~3H—甲氧补骨脂素被用于透皮机理研究,荧光显微镜观察法表明:①给予脂质体、脂质体凝胶、搽剂、凝胶4种8-MOP制剂的豚鼠皮脂腺较明显,且成团分布,皮脂腺官腔、腺泡扩大,其中以脂质体凝胶和凝胶组最明显;②正常组、PBS组表皮细胞密集、排列整齐,而脂质体组、脂质体凝胶组、搽剂组表皮细胞排列紊乱,细胞间隙有
Vitiligo is a condition in which the tyrosine system in pigment cells are destroyed , resulting in irregularly shaped white patches on the skin. We studied the preparation craft, skin-targetting property, the quality standard and the initial stability of 8-methoxyposoralen (MOP) liposome and liposomal gel, in order to prepare a skin targetting liposome gel of MOP with more effective and fewer side effects. We also studied the mechanic and influencing factors of skin-targetting property, aiming to prepare a more effective Vitiligo drug with fewer side effects.Film-supersonic method was better than antiphase evaporating method, film-dispersion method , and aether infusion method in preparating MOP liposome, which was evaluated with envelopment efficiency (EE) and mean diameter. High pressure homogeneous method was first used in preparating MOP nanoliposome by homogenizing 6 times at 1000 bar pressure. Carbopol 940 was used as matrix in preparating MOP liposome gel, in which liposome was added before the gel formed. The quality of liposome and liposomal gel were evaluated in general.The liposome had a fine-looking outward appearance, narrow size-distribution under the transmission electron microscope (TEM) and in laser diffraction particle size analyzer. The mean diameter of liposome and liposomal gel was 280 nm and 332.8 nm respectively. The appearance of liposomal gel was characterized by freeze-etching technique. The EE of five batches of liposomes was 94.2%, 97.5%, 93.4%, 95.3% and 95.7% respectively. That of the two batches of liposomal gel was 91.5% and 87.7% respectively, which were determined with sephadex column-HPLC method. The content of MOP in the liposome and liposomal gel was 2.96, 2.82, 2.88, 3.03, S.llmg.mL~(-1) and 1.82, 1.93 mg. g~(-1) respectively. The liposome and the liposomal gel were stable after 90 days at 4℃ or 25 ℃, but they were unstable at high temperature (40℃) and under strong light (4000~5000LX) condition.The transdermal permeability in vitro of MOP liposome through mouse skin was tested. The drug detained in the skin was 2.7 times and
试验以包封率、平均粒径为主要考察指标,分别采用逆相蒸发法、干膜振荡分散法、薄膜-超声法、乙醚注入法制备MOP脂质体进行比较,确定薄膜-超声法为佳,并确定高压均质法制备纳米脂质体的工艺为1000bar均质6次。以卡波姆(CP)为基质,采用基质形成前加入脂质体的方法制备了脂质体凝胶,并对脂质体、脂质体凝胶的质量进行全面评价,认为脂质体及其凝胶制备工艺基本稳定、质量可控,用负染法、扫描电镜法对脂质体形态进行表征,脂质体颗粒圆整、均匀,并用冷冻蚀刻法对脂质体凝胶形态进行表征。激光散射法测定脂质体及脂质体凝胶的平均粒径分别为280nm、332nm,葡聚糖凝胶柱法测定5批供试样品包封率分别为94.2%、97.5%、93.4%、95.3%、95.7%,含量分别为2.96、2.82、2.88、3.03、3.11mg·mL~(-1)。2批脂质体凝胶的包封率分别为91.5%、87.7%,含量为1.82、1.93mg·g~(-1)。脂质体稳定性研究表明脂质体及其凝胶剂在4℃、25℃贮存90天稳定,对高温及强光不稳定。
体外透皮实验表明滞留于皮肤内的药量MOP脂质体比MOP搽剂高2.7倍,较脂质体凝胶高1.5倍,MOP脂质体凝胶比MOP搽剂高1.7倍;电镜观察表明脂质体不是直接透过皮肤进入血液,而是在皮肤中代谢、释放药物,然后药物再渗透、穿过皮肤。
~3H—甲氧补骨脂素被用于透皮机理研究,荧光显微镜观察法表明:①给予脂质体、脂质体凝胶、搽剂、凝胶4种8-MOP制剂的豚鼠皮脂腺较明显,且成团分布,皮脂腺官腔、腺泡扩大,其中以脂质体凝胶和凝胶组最明显;②正常组、PBS组表皮细胞密集、排列整齐,而脂质体组、脂质体凝胶组、搽剂组表皮细胞排列紊乱,细胞间隙有
Vitiligo is a condition in which the tyrosine system in pigment cells are destroyed , resulting in irregularly shaped white patches on the skin. We studied the preparation craft, skin-targetting property, the quality standard and the initial stability of 8-methoxyposoralen (MOP) liposome and liposomal gel, in order to prepare a skin targetting liposome gel of MOP with more effective and fewer side effects. We also studied the mechanic and influencing factors of skin-targetting property, aiming to prepare a more effective Vitiligo drug with fewer side effects.Film-supersonic method was better than antiphase evaporating method, film-dispersion method , and aether infusion method in preparating MOP liposome, which was evaluated with envelopment efficiency (EE) and mean diameter. High pressure homogeneous method was first used in preparating MOP nanoliposome by homogenizing 6 times at 1000 bar pressure. Carbopol 940 was used as matrix in preparating MOP liposome gel, in which liposome was added before the gel formed. The quality of liposome and liposomal gel were evaluated in general.The liposome had a fine-looking outward appearance, narrow size-distribution under the transmission electron microscope (TEM) and in laser diffraction particle size analyzer. The mean diameter of liposome and liposomal gel was 280 nm and 332.8 nm respectively. The appearance of liposomal gel was characterized by freeze-etching technique. The EE of five batches of liposomes was 94.2%, 97.5%, 93.4%, 95.3% and 95.7% respectively. That of the two batches of liposomal gel was 91.5% and 87.7% respectively, which were determined with sephadex column-HPLC method. The content of MOP in the liposome and liposomal gel was 2.96, 2.82, 2.88, 3.03, S.llmg.mL~(-1) and 1.82, 1.93 mg. g~(-1) respectively. The liposome and the liposomal gel were stable after 90 days at 4℃ or 25 ℃, but they were unstable at high temperature (40℃) and under strong light (4000~5000LX) condition.The transdermal permeability in vitro of MOP liposome through mouse skin was tested. The drug detained in the skin was 2.7 times and
引文
[1] 高天文.挑战白癜风治疗[J].第四军医大学学报,2001;22(24):3122
[2] 许定安.一例白癜风患者家系的遗传分析[J].临床皮肤科杂志,1994;23(5):257.
[3] 张淑妍.银屑病愈后继发白癜风及家族性白癜风2例[J].中国皮肤性病学杂志,1996;10(2):96.
[4] (刘承煌译).白癜风患者的免疫状态[J].国外医学皮肤性病学分册,1995;21(6):369.
[5] 谢忠,季素珍,沈丽玉,等.白癜风患者血清中抗黑素细胞自身抗体与临床发病的关系[J].中华皮肤科杂志,1995;28(4):232.
[6] 谢忠,季素珍,沈丽玉,等.白癜风抗原的辨认[J].中华皮肤科杂志,1995;28(4):226.
[7] 季素珍,施惠娟,沈丽玉.白癜风患者血清中可溶性白介素2受体的检测[J].中国皮肤性病学杂志,1999;13(1):15.
[8] 杜娟,朱铁君.白癜风的超微结构变化[J].临床皮肤科杂志,1996;25(4):238.
[9] 杜娟,朱铁君.寻常型进行期白癜风的超微结构变化[J].中华皮肤科杂志,1996;29(4):240.
[10] 石得仁,普雄明,谢凤莲.两种类型白癜风患者郎格罕细胞电镜观察[J].临床皮肤科杂志,1996;25(3):129.
[11] 杜娟,朱铁君.白癜风患者郎格罕细胞光、电镜研究[J].临床皮肤科杂志,1996;25(5):273.
[12] 漆军,许锐,庄国康,等.散发型白癜风的组织病理变化[J].中国皮肤性病学杂志,1996;10(3):145.
[13] 黄惠明.人头发钴含量与白癜风[J].江西医药,1995;30(1):6.
[14] 涂彩霞,陈海波,林熙然.白癜风患者血清及皮肤组织液硒元素测定[J].中华皮肤科杂志,1999;32(3):184.
[15] 方跃明,王金良.现代实用皮肤性病学及美容学[M].上海:华东理工大学出版社,1995.367.
[16] 文京华.国内外白癜风治疗方法综述[J].皮肤病与性病,2002;24(2):23
[17] 赵达明.白癜风治疗现状和进展[J].人民军医,2002;45(8):26
[18] 史月君,孙艳.复方补骨脂搽剂治疗白癜风[J].中国皮肤病研究,2000;16(2):821
[19] 何文,雷嘉川,冯晓东,等.复方补骨脂素凝胶的制备及临床疗效观察[J].中国医院药学杂志,2001;21(11):566
[2] 许定安.一例白癜风患者家系的遗传分析[J].临床皮肤科杂志,1994;23(5):257.
[3] 张淑妍.银屑病愈后继发白癜风及家族性白癜风2例[J].中国皮肤性病学杂志,1996;10(2):96.
[4] (刘承煌译).白癜风患者的免疫状态[J].国外医学皮肤性病学分册,1995;21(6):369.
[5] 谢忠,季素珍,沈丽玉,等.白癜风患者血清中抗黑素细胞自身抗体与临床发病的关系[J].中华皮肤科杂志,1995;28(4):232.
[6] 谢忠,季素珍,沈丽玉,等.白癜风抗原的辨认[J].中华皮肤科杂志,1995;28(4):226.
[7] 季素珍,施惠娟,沈丽玉.白癜风患者血清中可溶性白介素2受体的检测[J].中国皮肤性病学杂志,1999;13(1):15.
[8] 杜娟,朱铁君.白癜风的超微结构变化[J].临床皮肤科杂志,1996;25(4):238.
[9] 杜娟,朱铁君.寻常型进行期白癜风的超微结构变化[J].中华皮肤科杂志,1996;29(4):240.
[10] 石得仁,普雄明,谢凤莲.两种类型白癜风患者郎格罕细胞电镜观察[J].临床皮肤科杂志,1996;25(3):129.
[11] 杜娟,朱铁君.白癜风患者郎格罕细胞光、电镜研究[J].临床皮肤科杂志,1996;25(5):273.
[12] 漆军,许锐,庄国康,等.散发型白癜风的组织病理变化[J].中国皮肤性病学杂志,1996;10(3):145.
[13] 黄惠明.人头发钴含量与白癜风[J].江西医药,1995;30(1):6.
[14] 涂彩霞,陈海波,林熙然.白癜风患者血清及皮肤组织液硒元素测定[J].中华皮肤科杂志,1999;32(3):184.
[15] 方跃明,王金良.现代实用皮肤性病学及美容学[M].上海:华东理工大学出版社,1995.367.
[16] 文京华.国内外白癜风治疗方法综述[J].皮肤病与性病,2002;24(2):23
[17] 赵达明.白癜风治疗现状和进展[J].人民军医,2002;45(8):26
[18] 史月君,孙艳.复方补骨脂搽剂治疗白癜风[J].中国皮肤病研究,2000;16(2):821
[19] 何文,雷嘉川,冯晓东,等.复方补骨脂素凝胶的制备及临床疗效观察[J].中国医院药学杂志,2001;21(11):566