替加氟前体脂质体软胶囊的研制
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摘要
目的:本课题将替加氟新型前体脂质体制成软胶囊,提高前体脂质体的稳定性,方便临床使用。主要完成制剂处方组成及工艺研究,质量标准的研究和初步稳定性考察。
方法:(1) 以丙二醛(MDA)为指标,筛选抗氧化剂Vit-E的用量。(2) 以柠檬黄为指示剂测定囊皮的溶出速率,采用单因素试验设计与均匀试验设计优选出软胶囊囊皮处方。(3) 用HPLC方法测定大豆磷脂的主要成分磷脂酰胆碱(PC),磷脂酰乙醇胺(PE)。(4)根据质量控制要求建立替加氟前体脂质体软胶囊的质量标准。(5) 根据中国药典2000版药物稳定性试验指导原则,进行加速试验与长期试验。
结果:(1) 抗氧化剂Vit-E(含量≥70%)的用量为6%。(2)优选出胶皮处方:甘油:明胶=1:0.847,PEG400:明胶=1:0.0586,TiO_2的用量在0.3%～4.0%之间对囊皮溶出速率无影响。(3) 用HPLC方法同时检测了大豆磷脂中的PC与PE的含量。经方法学验证,所确定的检测方法稳定可靠,重现性好。(4) 建立的质量标准可有效监测软胶囊及前体脂质体的质量。(5) 初步稳定性试验各项考察指标均未见明显变化。
结论:(1) 优化处方组成合理,工艺稳定。可用于工业化生产。(2) 质量标准可行,制剂质量可控。(3) 制剂的初步稳定性良好,在解决脂质体稳定性方面作了新的尝试。
Objective: The purpose of research is to make the soft capsules of tegafur proliposome, improving the stability of proliposome and convenienting the application for clinic. The projects include the screening of optimizing prescriptions and producing process of the tegafur proliposome soft capsules, building the quality criterion and to study the initial stage stability of the soft capsules.Methods: (1) According to the product of malonaldehyde (MDA), the quantity of Vit-E as oxidation inhibitor was selected. (2) The method with Tartrazine as indicator was used to determine the dissolution rates of the soft gelatin capsule shell. To optimize the shell compositions by the mono factor trail and the method of design by uniform distribution. (3) The HPLC method was applied to determinate the concentration of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in soybean phospholipid. (4) According to the requirement of building the criterion, the quality criterion of the tegafur proliposome sofe capsule was established. (5) Accelerating test and long-term test was studied according to Chinese Pharmacopeia (Edition 2000).Results: (1) the quantity of Vit-E as oxidation inhibitor (contents≥ 70%) is 6%. (2) The optimizing prescription is that glycerin/gelatin =0.847, PEG400/gelatin=0.0586, the quantity of TiO2 from 0.3%~4% is not interfere with the dissolution rate of capsule shell. (3) The PC and PE can be detected at the same HPLC condition. The method is sensitive,
accurate and reproducible. The recovery and precision of the method satisfied the requirements. (4) The quality criterion of the tegafur proliposome soft capsules can practicably control the quality. (5) The indexes were not obviously change in initial stage research.Conclusions: (1) Verified experiment made it known that the better proscription is reasonable and stable. (2) The quality control method is stable and has a good repeatability and can be used to control the quality of the proliposome soft capsules. (3) The tegafur proliposome soft capsule was stable under the different test condition in initial stage research.
方法:(1) 以丙二醛(MDA)为指标,筛选抗氧化剂Vit-E的用量。(2) 以柠檬黄为指示剂测定囊皮的溶出速率,采用单因素试验设计与均匀试验设计优选出软胶囊囊皮处方。(3) 用HPLC方法测定大豆磷脂的主要成分磷脂酰胆碱(PC),磷脂酰乙醇胺(PE)。(4)根据质量控制要求建立替加氟前体脂质体软胶囊的质量标准。(5) 根据中国药典2000版药物稳定性试验指导原则,进行加速试验与长期试验。
结果:(1) 抗氧化剂Vit-E(含量≥70%)的用量为6%。(2)优选出胶皮处方:甘油:明胶=1:0.847,PEG400:明胶=1:0.0586,TiO_2的用量在0.3%～4.0%之间对囊皮溶出速率无影响。(3) 用HPLC方法同时检测了大豆磷脂中的PC与PE的含量。经方法学验证,所确定的检测方法稳定可靠,重现性好。(4) 建立的质量标准可有效监测软胶囊及前体脂质体的质量。(5) 初步稳定性试验各项考察指标均未见明显变化。
结论:(1) 优化处方组成合理,工艺稳定。可用于工业化生产。(2) 质量标准可行,制剂质量可控。(3) 制剂的初步稳定性良好,在解决脂质体稳定性方面作了新的尝试。
Objective: The purpose of research is to make the soft capsules of tegafur proliposome, improving the stability of proliposome and convenienting the application for clinic. The projects include the screening of optimizing prescriptions and producing process of the tegafur proliposome soft capsules, building the quality criterion and to study the initial stage stability of the soft capsules.Methods: (1) According to the product of malonaldehyde (MDA), the quantity of Vit-E as oxidation inhibitor was selected. (2) The method with Tartrazine as indicator was used to determine the dissolution rates of the soft gelatin capsule shell. To optimize the shell compositions by the mono factor trail and the method of design by uniform distribution. (3) The HPLC method was applied to determinate the concentration of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in soybean phospholipid. (4) According to the requirement of building the criterion, the quality criterion of the tegafur proliposome sofe capsule was established. (5) Accelerating test and long-term test was studied according to Chinese Pharmacopeia (Edition 2000).Results: (1) the quantity of Vit-E as oxidation inhibitor (contents≥ 70%) is 6%. (2) The optimizing prescription is that glycerin/gelatin =0.847, PEG400/gelatin=0.0586, the quantity of TiO2 from 0.3%~4% is not interfere with the dissolution rate of capsule shell. (3) The PC and PE can be detected at the same HPLC condition. The method is sensitive,
accurate and reproducible. The recovery and precision of the method satisfied the requirements. (4) The quality criterion of the tegafur proliposome soft capsules can practicably control the quality. (5) The indexes were not obviously change in initial stage research.Conclusions: (1) Verified experiment made it known that the better proscription is reasonable and stable. (2) The quality control method is stable and has a good repeatability and can be used to control the quality of the proliposome soft capsules. (3) The tegafur proliposome soft capsule was stable under the different test condition in initial stage research.
引文
[1] 朱盛山.药物新剂型[M].化学工业出版社,2003:408
[2] 崔福德.药剂学(第五版)[M].人民卫生出版社,2004:388
[3] 黄敏,张钧寿,谢跃进.软胶囊剂稳定性研究进展[J].中国医药工业杂志,2000,31(3):137-140
[4] 王长虹,孙殿甲.脂质体的物理化学稳定性研究进展[J].中国药学杂志,1998,33(2):65-68
[5] 刘元法,王兴国.大豆磷脂组成与特性[J].粮食与油脂,2000,(3):11-14
[6] G. E. Dobretsov, T. A. Borschevskava, V. A. Petrov, et al. The increase of phospholipid bilayer rigidity after lipid peroxidation [J]. FEBS Lett, 1977, 84: 125-128.
[7] R. C. Bruch, W. S. Thayer. Differential effect of lipid peroxidation on membrane fluidity as determined by electron spin resonance probes [J]. Biochim Biophys Acta, 1983, 733: 216-222.
[8] 林如,王齐放,黄玉萍等.大豆磷脂及抗氧化动力学规律的研究[J].沈阳药学院学报,1994,11(4):235-238
[9] 何新霞,徐丽珊,杨玲.大豆磷脂胶丸稳定性初步观察[J].中国药学杂志,1999,34(8):538-540
[10] 马辰,段宏瑾.大豆磷脂中磷脂类成分的含量测定[J].中国中药杂志,1999,24(11)671-672
[11] John C. Dittmer, Robert L. Lester. A simple, specific spray for the detection of phospholipids on thin-layer chromatograms [J]. J Lipid Res, 1961, 5: 126～127.
[12] W. D. Skidmore, C. Entenman. Two-dimensional thin-layer chromatography of rat liver phosphatides[J]. J Lipid Res, 1962, 3(4): 471-475.
[13] 吴道澄,席晓莉,胡佳蕙.分光光度法测定阿霉素脂质体中的磷脂含量[J].解放军药学学报,1999,15(3):48-50
[14] 林筱琦,王建华,温浩等.阿苯达唑脂质体中磷脂含量测定[J].新疆医科大学学报,2002,25(2)119-120
[15] J. A. Singleton. Enrichment of phospholipids from neutral lipids in peanut oil by high-performance liquid chromatography[J]. JAOCS, 1993, 70(6): 637-638.
[16] Duk Hui Kang, Kyung Ho Row. Fractionation of soybean phospholipids by preparative high performance liquid chromatography with sorbents of various particle size[J]. J Chromatogr A, 2002, 949: 217-223.
[17] Terry L. Kaduce, Kurt C. Norton, Arthur A. Spector. A rapid, isocratic method for phospholipid separation by high-performance liquid chromatography[J]. J Lipid Res, 1983, 24: 1398-1403.
[18] E. H. Farmer, D. A. Sutton. The course of autoxidation reactions in polyisoprenes and allied compounds. Part Ⅳ. The isolation and constitution of photochemically-formed methyl oleate peroxide[J]. J Chem Soc, 1943: 119-122.
[19] L. K. Dahle, E. G. Hill, R. T. Holman. The thiobarbituric acid reaction and the autoxidations of polyunsaturated fatty acid methyl esters[J]. Arch Biochem Biophys, 1962, 98: 253-261.
[20] H. H. Draper, M. Hadley. Malondialdehyde determination as index of lipid peroxidation[J]. Methods Enzymol, 1990, 186: 421-431.
[21] 翁帼英,陈明非,王玲玲.脂质体中磷脂的氧化产物与溶血的关系[J].生物化学与生物物理进展,1990,17(1):76-77
[22] 陈骐,黄芸,顾学裘.喷雾干燥法制各前体脂质体的基础研究[J].沈阳药科大学学报,1997,14(3):166-169
[23] 莫凤奎,朱澄云,黄松鹤.维生素E和维生素C对大豆磷脂脂质体的抗氧化作用[J].中国药物化学杂志,1997,7(2);121-124
[24] Manabu K, Keizo I, Shoshichi N. Effect of ferrous ion and scorbateinduced lipid peroxidation on liposomal membranes[J]. Biochem Biophys Acta, 1981, 646: 169-178
[25] F. S. Hom, S. A. Veresh. Soft, et al. Gelation Capsules Ⅰ: Factors Affecting Capsule Shell Dissolution Rate[J]. J Pharm Sci, 1973, 62(6): 1001-1006.
[26] 唐星,何仲贵,杨杨.明胶软胶囊胶壳处方因素对囊壳溶解性能的影响[J].中国药学杂志,1999,34(1):28-30.
[27] 李瑞雪.均匀设计在药物研究开发中的应用[M].四川科学技术出版社,2000
[28] Laurent M, Benoit A, Pierre C, et al. Determination of Phospholipids from Pulmonary Surfactant Using an On-Line Coupled Silica/ ReversedPhase High-Performance Liquid Chromatography System[J]. Anal Chem, 1992, 84: 371-379
[29] W. S. M. Geurts, W. M. A. Hax, R. A. Demel, et al. High performance liquid chromatographic separation and direct ultraviolet detection of phospholipids[J]. Biochimia Biophysica Acta, 1977, (486): 524-530
[30] Mustafa Grit, Daan J. A. Crommelin. Determination of phosphatidylcholine, phosphatidylglycerol and their lyso forms from liposome dispersions by high-performance liquid chromatography using highsensitivity refractive index detection[J]. J Chromatogr, 1991, 585: 239-246.
[31] J. Terao, I. Asano, S. Matsushita. Preparation of Hydroperoxy and hydroxy derivatives of rat liver phosphatidycholine and phosphatidyethanolamine[J]. Lipids, 1985, 20(5): 312-317.
[32] 吴骏,朱家璧.阿昔洛韦脂质体的制备和稳定性的初步考察[J].药学学报,2003,38(7):552-554
[33] 魏树礼,张强.生物药剂学与药物动力学(第二版)[M].北京大学医学出版社,2004:408-410
[34] 朱盛山.药物新剂型[M].化学工业出版社,2003:445
[35] 王建华,林筱琦,陈迹等.阿苯达唑脂质体口服液质量控制中薄层层析条件的筛选与应用[J].新疆医科大学学报,2001,24(3):192-194
[36] 吴道澄,席晓莉.多柔比星磷脂复合材料脂质体的制备[J].中国医药工业杂志,1999,30(1):15-18
[37] 胥传来,乐国伟,姚惠源.不同组分对PST脂质体化学稳定性的影响[J].中国油脂,2002,27(3):74-75
[38] S. Shi-hua Chen, Anne Y. Kou. Improved procedure for the separation of phospholipids by high-performance liquid chromatography[J]. J Chromatogr, 1982, 227: 25-31.
[39] W. Jeffrey Hurst, Robert A. martin, JR. The analysis of phospholipids in soy lecithin by HPLC[J]. JAOCS, 1984, 61(9): 1462-1463.
[40] 张奇,邓英杰.冻融法制备 5-氟脲嘧啶脂质体及其稳定性考察[J].沈阳药科大学学报,2000,17(2):87-89
[41] 戴淑真,曾骏文,王子亮等.哌仑西平脂质体的研制及评介[J].中山大学学报(医学科学版),2004,25,(3):217-220
[42] 王九成,惠民权,焦亚奇等.三瓶装阿霉素脂质体注射液制剂研究[J].世界最新医学信息文摘,2003,2(4):746-749
[43] 肖超菊,齐宪荣,艾尼瓦尔等.顺铂缓释多囊脂质体的制备和体外释放性能研究[J].药学学报,2003,38(2):133-137
[44] 张奇,邓英杰,罗国安.均匀设计制备 5-氟脲嘧啶脂质体及其稳定性[J].北京大学学报(医学版),2002,34(1):64-67
[45] 储茂泉,古宏晨,刘国杰.丹参酮脂质体的制备及其稳定性[J].中国医药工业杂志,2001,32(9):400-404
[46] 李晓光,翟所迪.甘草酸单铵盐脂质体的试制及稳定性研究[J].中国医药工业杂志,2004,35(4):222-224
[47] M. Joshi, A. Misra. Dry power inhalation of liposomal ketotifen fumarate: formulation and characterization[J]. Int J Pharm, 2001, 223: 15-27
[2] 崔福德.药剂学(第五版)[M].人民卫生出版社,2004:388
[3] 黄敏,张钧寿,谢跃进.软胶囊剂稳定性研究进展[J].中国医药工业杂志,2000,31(3):137-140
[4] 王长虹,孙殿甲.脂质体的物理化学稳定性研究进展[J].中国药学杂志,1998,33(2):65-68
[5] 刘元法,王兴国.大豆磷脂组成与特性[J].粮食与油脂,2000,(3):11-14
[6] G. E. Dobretsov, T. A. Borschevskava, V. A. Petrov, et al. The increase of phospholipid bilayer rigidity after lipid peroxidation [J]. FEBS Lett, 1977, 84: 125-128.
[7] R. C. Bruch, W. S. Thayer. Differential effect of lipid peroxidation on membrane fluidity as determined by electron spin resonance probes [J]. Biochim Biophys Acta, 1983, 733: 216-222.
[8] 林如,王齐放,黄玉萍等.大豆磷脂及抗氧化动力学规律的研究[J].沈阳药学院学报,1994,11(4):235-238
[9] 何新霞,徐丽珊,杨玲.大豆磷脂胶丸稳定性初步观察[J].中国药学杂志,1999,34(8):538-540
[10] 马辰,段宏瑾.大豆磷脂中磷脂类成分的含量测定[J].中国中药杂志,1999,24(11)671-672
[11] John C. Dittmer, Robert L. Lester. A simple, specific spray for the detection of phospholipids on thin-layer chromatograms [J]. J Lipid Res, 1961, 5: 126～127.
[12] W. D. Skidmore, C. Entenman. Two-dimensional thin-layer chromatography of rat liver phosphatides[J]. J Lipid Res, 1962, 3(4): 471-475.
[13] 吴道澄,席晓莉,胡佳蕙.分光光度法测定阿霉素脂质体中的磷脂含量[J].解放军药学学报,1999,15(3):48-50
[14] 林筱琦,王建华,温浩等.阿苯达唑脂质体中磷脂含量测定[J].新疆医科大学学报,2002,25(2)119-120
[15] J. A. Singleton. Enrichment of phospholipids from neutral lipids in peanut oil by high-performance liquid chromatography[J]. JAOCS, 1993, 70(6): 637-638.
[16] Duk Hui Kang, Kyung Ho Row. Fractionation of soybean phospholipids by preparative high performance liquid chromatography with sorbents of various particle size[J]. J Chromatogr A, 2002, 949: 217-223.
[17] Terry L. Kaduce, Kurt C. Norton, Arthur A. Spector. A rapid, isocratic method for phospholipid separation by high-performance liquid chromatography[J]. J Lipid Res, 1983, 24: 1398-1403.
[18] E. H. Farmer, D. A. Sutton. The course of autoxidation reactions in polyisoprenes and allied compounds. Part Ⅳ. The isolation and constitution of photochemically-formed methyl oleate peroxide[J]. J Chem Soc, 1943: 119-122.
[19] L. K. Dahle, E. G. Hill, R. T. Holman. The thiobarbituric acid reaction and the autoxidations of polyunsaturated fatty acid methyl esters[J]. Arch Biochem Biophys, 1962, 98: 253-261.
[20] H. H. Draper, M. Hadley. Malondialdehyde determination as index of lipid peroxidation[J]. Methods Enzymol, 1990, 186: 421-431.
[21] 翁帼英,陈明非,王玲玲.脂质体中磷脂的氧化产物与溶血的关系[J].生物化学与生物物理进展,1990,17(1):76-77
[22] 陈骐,黄芸,顾学裘.喷雾干燥法制各前体脂质体的基础研究[J].沈阳药科大学学报,1997,14(3):166-169
[23] 莫凤奎,朱澄云,黄松鹤.维生素E和维生素C对大豆磷脂脂质体的抗氧化作用[J].中国药物化学杂志,1997,7(2);121-124
[24] Manabu K, Keizo I, Shoshichi N. Effect of ferrous ion and scorbateinduced lipid peroxidation on liposomal membranes[J]. Biochem Biophys Acta, 1981, 646: 169-178
[25] F. S. Hom, S. A. Veresh. Soft, et al. Gelation Capsules Ⅰ: Factors Affecting Capsule Shell Dissolution Rate[J]. J Pharm Sci, 1973, 62(6): 1001-1006.
[26] 唐星,何仲贵,杨杨.明胶软胶囊胶壳处方因素对囊壳溶解性能的影响[J].中国药学杂志,1999,34(1):28-30.
[27] 李瑞雪.均匀设计在药物研究开发中的应用[M].四川科学技术出版社,2000
[28] Laurent M, Benoit A, Pierre C, et al. Determination of Phospholipids from Pulmonary Surfactant Using an On-Line Coupled Silica/ ReversedPhase High-Performance Liquid Chromatography System[J]. Anal Chem, 1992, 84: 371-379
[29] W. S. M. Geurts, W. M. A. Hax, R. A. Demel, et al. High performance liquid chromatographic separation and direct ultraviolet detection of phospholipids[J]. Biochimia Biophysica Acta, 1977, (486): 524-530
[30] Mustafa Grit, Daan J. A. Crommelin. Determination of phosphatidylcholine, phosphatidylglycerol and their lyso forms from liposome dispersions by high-performance liquid chromatography using highsensitivity refractive index detection[J]. J Chromatogr, 1991, 585: 239-246.
[31] J. Terao, I. Asano, S. Matsushita. Preparation of Hydroperoxy and hydroxy derivatives of rat liver phosphatidycholine and phosphatidyethanolamine[J]. Lipids, 1985, 20(5): 312-317.
[32] 吴骏,朱家璧.阿昔洛韦脂质体的制备和稳定性的初步考察[J].药学学报,2003,38(7):552-554
[33] 魏树礼,张强.生物药剂学与药物动力学(第二版)[M].北京大学医学出版社,2004:408-410
[34] 朱盛山.药物新剂型[M].化学工业出版社,2003:445
[35] 王建华,林筱琦,陈迹等.阿苯达唑脂质体口服液质量控制中薄层层析条件的筛选与应用[J].新疆医科大学学报,2001,24(3):192-194
[36] 吴道澄,席晓莉.多柔比星磷脂复合材料脂质体的制备[J].中国医药工业杂志,1999,30(1):15-18
[37] 胥传来,乐国伟,姚惠源.不同组分对PST脂质体化学稳定性的影响[J].中国油脂,2002,27(3):74-75
[38] S. Shi-hua Chen, Anne Y. Kou. Improved procedure for the separation of phospholipids by high-performance liquid chromatography[J]. J Chromatogr, 1982, 227: 25-31.
[39] W. Jeffrey Hurst, Robert A. martin, JR. The analysis of phospholipids in soy lecithin by HPLC[J]. JAOCS, 1984, 61(9): 1462-1463.
[40] 张奇,邓英杰.冻融法制备 5-氟脲嘧啶脂质体及其稳定性考察[J].沈阳药科大学学报,2000,17(2):87-89
[41] 戴淑真,曾骏文,王子亮等.哌仑西平脂质体的研制及评介[J].中山大学学报(医学科学版),2004,25,(3):217-220
[42] 王九成,惠民权,焦亚奇等.三瓶装阿霉素脂质体注射液制剂研究[J].世界最新医学信息文摘,2003,2(4):746-749
[43] 肖超菊,齐宪荣,艾尼瓦尔等.顺铂缓释多囊脂质体的制备和体外释放性能研究[J].药学学报,2003,38(2):133-137
[44] 张奇,邓英杰,罗国安.均匀设计制备 5-氟脲嘧啶脂质体及其稳定性[J].北京大学学报(医学版),2002,34(1):64-67
[45] 储茂泉,古宏晨,刘国杰.丹参酮脂质体的制备及其稳定性[J].中国医药工业杂志,2001,32(9):400-404
[46] 李晓光,翟所迪.甘草酸单铵盐脂质体的试制及稳定性研究[J].中国医药工业杂志,2004,35(4):222-224
[47] M. Joshi, A. Misra. Dry power inhalation of liposomal ketotifen fumarate: formulation and characterization[J]. Int J Pharm, 2001, 223: 15-27