左卡尼汀温敏原位凝胶的制备及质量评价
|
摘要
本文以左卡尼汀为模型药物,制备了一种用于治疗初发型干眼病的新型制剂-左卡尼汀温敏原位凝胶。以泊洛沙姆407和泊洛沙姆188为凝胶基质,透明质酸钠和羧甲基纤维素钠为生物黏附性材料,采用转子法测定胶凝温度,星点设计-效应面法优化处方;并对最优处方的pH值、黏度值及胶凝温度等进行测定,透析膜透过法考察制剂体外释放,裂隙灯显微镜观察制剂在兔眼表的滞留性。结果表明,原位凝胶的最优处方中,泊洛沙姆407和泊洛沙姆188用量分别为20.81%和3.46%,透明质酸钠为0.02%,羧甲基纤维素钠为0.10%;室温下pH值为6.90±0.06,黏度在27℃开始发生突跃,泪液稀释前后的胶凝温度分别为(26.37±0.06)℃和(33.57±0.21)℃;体外单位面积释放速率在前240 min内低于溶液剂(P<0.05), 600 min时累积释放度可达80%以上;原位凝胶制剂滴眼给药后,在家兔眼表的滞留时间为25 min左右,约为溶液剂的5倍。动物实验按照国际动物实验指导原则进行并获得北京中医药大学实验动物中心伦理学委员会批准。本文所制得的左卡尼汀温敏原位凝胶性状良好,具有缓释特性并显著提高了药物在家兔眼表的滞留时间。
In this paper, a new type of preparation for treatment of initial dry eye disease, thermosensitive in situ gel, was prepared using levocarnitine as a model drug. Poloxamer 407 and poloxamer 188 were used as the gel matrix, and sodium hyaluronate and sodium carboxymethylcellulose were used as bioadhesive materials.Gelation temperature was determined by a rotor method and the prescription was optimized by central composite design-response surface methodology. The pH value, viscosity value and gelation temperature of the optimal prescription were measured. The release of the drug in vitro was examined by dialysis membrane permeation, and retention time of the thermosensitive in situ gel preparation on the rabbit's ocular surface was observed by a slit lamp microscope. The results showed that the dosage of the poloxamer 407 and poloxamer 188 were 20.81% and3.46%, respectively, and sodium hyaluronate was 0.02%, sodium carboxymethyl cellulose was 0.10% of the optimal formulation of levocarnitine thermosensitive in situ gel. The pH value was 6.90 ± 0.06 at room temperature and the viscosity value started to rise sharply at 27 ℃ of the optimal formulation. The gelation temperature of the optimal preparation before and after dilution by simulated tear fluid were(26.37 ± 0.06) ℃ and(33.57 ± 0.21) ℃, respec‐tively. In the first 240 min, in vitro release rate per unit area of levocarnitine thermosensitive in situ gel was lower than that of solution(P<0.05), and after 600 min, the cumulative release rate of levocarnitine thermosensitive in situ gel could reach more than 80%. The retention time of the thermosensitive in situ gel preparation on rabbit's ocular surface reached about 25 min, at least 5 times as much as that of the solution. The animal experiment was conducted following the National Institutes of Health Guidelines for the use of experimental animals, and approved by the Ethics Committee of the Experimental Animal Center of Beijing University of Chinese Medicine. The levocarnitine thermosensitive in situ gel showed good characteristics and sustained release property and significantly improved the retention time of the drug on the rabbit's ocular surface.
In this paper, a new type of preparation for treatment of initial dry eye disease, thermosensitive in situ gel, was prepared using levocarnitine as a model drug. Poloxamer 407 and poloxamer 188 were used as the gel matrix, and sodium hyaluronate and sodium carboxymethylcellulose were used as bioadhesive materials.Gelation temperature was determined by a rotor method and the prescription was optimized by central composite design-response surface methodology. The pH value, viscosity value and gelation temperature of the optimal prescription were measured. The release of the drug in vitro was examined by dialysis membrane permeation, and retention time of the thermosensitive in situ gel preparation on the rabbit's ocular surface was observed by a slit lamp microscope. The results showed that the dosage of the poloxamer 407 and poloxamer 188 were 20.81% and3.46%, respectively, and sodium hyaluronate was 0.02%, sodium carboxymethyl cellulose was 0.10% of the optimal formulation of levocarnitine thermosensitive in situ gel. The pH value was 6.90 ± 0.06 at room temperature and the viscosity value started to rise sharply at 27 ℃ of the optimal formulation. The gelation temperature of the optimal preparation before and after dilution by simulated tear fluid were(26.37 ± 0.06) ℃ and(33.57 ± 0.21) ℃, respec‐tively. In the first 240 min, in vitro release rate per unit area of levocarnitine thermosensitive in situ gel was lower than that of solution(P<0.05), and after 600 min, the cumulative release rate of levocarnitine thermosensitive in situ gel could reach more than 80%. The retention time of the thermosensitive in situ gel preparation on rabbit's ocular surface reached about 25 min, at least 5 times as much as that of the solution. The animal experiment was conducted following the National Institutes of Health Guidelines for the use of experimental animals, and approved by the Ethics Committee of the Experimental Animal Center of Beijing University of Chinese Medicine. The levocarnitine thermosensitive in situ gel showed good characteristics and sustained release property and significantly improved the retention time of the drug on the rabbit's ocular surface.
引文
[1] Khajavi N, Reinach PS, Skrzypski M, et al. L-Carnitine reduces in human conjunctival epithelial cells hypertonic-induced shrinkage through interacting with TRPV1 channels[J]. Cell Physiol Biochem,2014, 34:790-803.
[2] Hua X, Su ZT, Deng RZ, et al. Effects of L-camitine, erythritol and betaine on pro-inflammatory markers in primary human corneal epithelial cells exposed to hyperosmotic stress[J]. Curr Eye Res, 2015,40:657-667.
[3] Huo J. A Primary Study of L-Carnitine Protective Effect on Ocular Surface of Mouse Dry Eye Model Induced by Hyperosmolar Saline(左卡尼汀对高渗盐水诱导的小鼠干眼模型眼表保护作用的初步研究)[D]. Chongqing:Third Military Medical University, 2012.
[4] Jones L, Downie LE, Korb D, et al. TFOS DEWSⅡmanagement and therapy report[J]. Ocul Surf, 2017, 15:575-628.
[5] Moon JH, Kim KW, Moon NJ. Smartphone use is a risk factor for pediatric dry eye disease according to region and age:a case control study[J]. BMC Ophthalmol, 2016, 16:188.
[6] Baudouin C, Aragona P, Messmer EM, et al. Role of hyperosmolarity in the pathogenesis and management of dry eye disease:proceedings of the OCEAN Group Meeting[J]. Ocul Surf, 2013,11:246-258.
[7] Zhang Z, Li YH, Ding YL, et al. New progress in the pathogenesis and treatment of dry eyes[J]. Chin J Ophthalmol Med(中华眼科医学杂志),2014, 4:44-46.
[8] Pucker AD, Ng SM, Nichols JJ. Over the counter(OTC)artificial tear drops for dry eye syndrome[J]. Cochrane Database Syst Rev, 2016, 2:CD009729.
[9] Bron AJ, de Paiva CS, Chauhan SK, et al. TFOS DEWSⅡpathophysiology report[J]. Ocul Surf, 2017, 15:438-510.
[10] Steinfeld A, Lux A, Maier S, et al. Bioavailability of fluorescein from a new drug delivery system in human eyes[J]. Br J Ophthalmol, 2004, 88:48-53.
[11] Ao H, Bai JQ, Zhang YT, et al. Research progress in in-situ gel used in Chinese medicine targeting preparations[J]. China Pharm(中国药师),2017, 20:1283-1286.
[12] Bie HY,Wang HL,Wang YT. Optimization of azelastine hydro-chloride thermosensitive in situ gel eye drops by central composite design-response surface methodology[J]. Chin J New Drugs(中国新药杂志),2016, 25:954-960.
[13] Li LJ, Guo DD, Guo JG, et al. Thermosensitive in-situ forming gels for ophthalmic delivery of tea polyphenols[J]. J Drug Deliv Sci Technol, 2018, 46:243-250.
[14] Song LN, Li HR, Wang HY, et al. Preparation and properties of thermosensitive in situ gel for ophthalmic formulation containing pearl hydrolyzate[J]. Acta Pharm Sin(药学学报),2016,51:1622-1628.
[15] Yang HY, Xia JL, Lin SQ. Preparation of ophthalmic thermosensitive in situ gel of moxifloxacin hydrochloride and its release behavior in vitro[J]. Chin Pharm J(中国药学杂志),2014, 49:1827-1831.
[16] Chang XH, Yan XX, He X, et al. Preparation of ophthalmic thermosensitive in situ gel of azithromycin and the in vitro release mechanism[J]. Chin J New Drugs(中国新药杂志),2011,20:1577-1582.
[17] Zhang M, Li YY. Optimization of the rmosensitive in situ gel system containing ligustrazine hydrochloride for ocular use by central composite design-response surface methodology[J].Shandong Chem Ind(山东化工),2014, 43:15-18.
[18] Zhag JJ, Wang B. Preparation of thermosensitive in situ gel containing ganciclovir chitosan nanoparticle for ophthalmic use and study on its releasein vitro[J]. Prog Pharm Sci(药学进展),2013,37:522-527.
[19] Zhao W, Liu R, Sun L, et al. Effects of three kinds of cyclo-dextrin on in vitro corneal penetration of curcumin[J]. Chin Tradit Herb Drugs(中草药),2015, 46:201-206.
[20] Liang N. Study of diclofenac temperature sensitive hydrogel with bioadhesive eye[J]. Chin J Med Guide(中国医药导刊),2015, 17:177-179.
[21] Hao JF, Zhao XM, Wang JZ, et al. Optimization of thermosensitive in situ gel system containing berberine hydrochloride for ocular use[J]. Chin Tradit Herb Drugs(中草药),2010, 41:550-555.
[22] Hu J, Chen DW, Quan DQ. Rheological properties of poloxamer407 aqueous solutions[J]. Acta Pharm Sin(药学学报),2011,46:227-231.
[23] Xu JY, Yang HJ, Xiong X, et al. Research progression of several common temperature-sensitive in situ gel carriers[J]. Chin J Exp Tradit Med Formul(中国实验方剂学杂志),2011, 17:252-257.
[24] Yuan Y, Cui Y, Zhang L, et al. Thermosensitive and mucoadhesive in situ gel based on poloxamer as new carrier for rectal administration of nimesulide[J]. Int J Pharm, 2012, 430:114-119.
[25] Chen JS, Wang JX, Yi Y, et al. The research progress in hyaluronic acid[J]. China Biotechnol(中国生物工程杂志),2015,35:111-118.
[26] She YJ, Li JY, Xiao B, et al. Evaluation of a novel artificial tear in the prevention and treatment of dry eye in an animal model[J]. J Ocul Pharmacol Ther,2015, 31:525-530.
[27] Tian PC, Wang J. Preparation and ocular retention of aciclovir chitosan eye-drops[J]. J Guangdong Pharm Univ(广东药学学报),2013, 29:225-228.
[2] Hua X, Su ZT, Deng RZ, et al. Effects of L-camitine, erythritol and betaine on pro-inflammatory markers in primary human corneal epithelial cells exposed to hyperosmotic stress[J]. Curr Eye Res, 2015,40:657-667.
[3] Huo J. A Primary Study of L-Carnitine Protective Effect on Ocular Surface of Mouse Dry Eye Model Induced by Hyperosmolar Saline(左卡尼汀对高渗盐水诱导的小鼠干眼模型眼表保护作用的初步研究)[D]. Chongqing:Third Military Medical University, 2012.
[4] Jones L, Downie LE, Korb D, et al. TFOS DEWSⅡmanagement and therapy report[J]. Ocul Surf, 2017, 15:575-628.
[5] Moon JH, Kim KW, Moon NJ. Smartphone use is a risk factor for pediatric dry eye disease according to region and age:a case control study[J]. BMC Ophthalmol, 2016, 16:188.
[6] Baudouin C, Aragona P, Messmer EM, et al. Role of hyperosmolarity in the pathogenesis and management of dry eye disease:proceedings of the OCEAN Group Meeting[J]. Ocul Surf, 2013,11:246-258.
[7] Zhang Z, Li YH, Ding YL, et al. New progress in the pathogenesis and treatment of dry eyes[J]. Chin J Ophthalmol Med(中华眼科医学杂志),2014, 4:44-46.
[8] Pucker AD, Ng SM, Nichols JJ. Over the counter(OTC)artificial tear drops for dry eye syndrome[J]. Cochrane Database Syst Rev, 2016, 2:CD009729.
[9] Bron AJ, de Paiva CS, Chauhan SK, et al. TFOS DEWSⅡpathophysiology report[J]. Ocul Surf, 2017, 15:438-510.
[10] Steinfeld A, Lux A, Maier S, et al. Bioavailability of fluorescein from a new drug delivery system in human eyes[J]. Br J Ophthalmol, 2004, 88:48-53.
[11] Ao H, Bai JQ, Zhang YT, et al. Research progress in in-situ gel used in Chinese medicine targeting preparations[J]. China Pharm(中国药师),2017, 20:1283-1286.
[12] Bie HY,Wang HL,Wang YT. Optimization of azelastine hydro-chloride thermosensitive in situ gel eye drops by central composite design-response surface methodology[J]. Chin J New Drugs(中国新药杂志),2016, 25:954-960.
[13] Li LJ, Guo DD, Guo JG, et al. Thermosensitive in-situ forming gels for ophthalmic delivery of tea polyphenols[J]. J Drug Deliv Sci Technol, 2018, 46:243-250.
[14] Song LN, Li HR, Wang HY, et al. Preparation and properties of thermosensitive in situ gel for ophthalmic formulation containing pearl hydrolyzate[J]. Acta Pharm Sin(药学学报),2016,51:1622-1628.
[15] Yang HY, Xia JL, Lin SQ. Preparation of ophthalmic thermosensitive in situ gel of moxifloxacin hydrochloride and its release behavior in vitro[J]. Chin Pharm J(中国药学杂志),2014, 49:1827-1831.
[16] Chang XH, Yan XX, He X, et al. Preparation of ophthalmic thermosensitive in situ gel of azithromycin and the in vitro release mechanism[J]. Chin J New Drugs(中国新药杂志),2011,20:1577-1582.
[17] Zhang M, Li YY. Optimization of the rmosensitive in situ gel system containing ligustrazine hydrochloride for ocular use by central composite design-response surface methodology[J].Shandong Chem Ind(山东化工),2014, 43:15-18.
[18] Zhag JJ, Wang B. Preparation of thermosensitive in situ gel containing ganciclovir chitosan nanoparticle for ophthalmic use and study on its releasein vitro[J]. Prog Pharm Sci(药学进展),2013,37:522-527.
[19] Zhao W, Liu R, Sun L, et al. Effects of three kinds of cyclo-dextrin on in vitro corneal penetration of curcumin[J]. Chin Tradit Herb Drugs(中草药),2015, 46:201-206.
[20] Liang N. Study of diclofenac temperature sensitive hydrogel with bioadhesive eye[J]. Chin J Med Guide(中国医药导刊),2015, 17:177-179.
[21] Hao JF, Zhao XM, Wang JZ, et al. Optimization of thermosensitive in situ gel system containing berberine hydrochloride for ocular use[J]. Chin Tradit Herb Drugs(中草药),2010, 41:550-555.
[22] Hu J, Chen DW, Quan DQ. Rheological properties of poloxamer407 aqueous solutions[J]. Acta Pharm Sin(药学学报),2011,46:227-231.
[23] Xu JY, Yang HJ, Xiong X, et al. Research progression of several common temperature-sensitive in situ gel carriers[J]. Chin J Exp Tradit Med Formul(中国实验方剂学杂志),2011, 17:252-257.
[24] Yuan Y, Cui Y, Zhang L, et al. Thermosensitive and mucoadhesive in situ gel based on poloxamer as new carrier for rectal administration of nimesulide[J]. Int J Pharm, 2012, 430:114-119.
[25] Chen JS, Wang JX, Yi Y, et al. The research progress in hyaluronic acid[J]. China Biotechnol(中国生物工程杂志),2015,35:111-118.
[26] She YJ, Li JY, Xiao B, et al. Evaluation of a novel artificial tear in the prevention and treatment of dry eye in an animal model[J]. J Ocul Pharmacol Ther,2015, 31:525-530.
[27] Tian PC, Wang J. Preparation and ocular retention of aciclovir chitosan eye-drops[J]. J Guangdong Pharm Univ(广东药学学报),2013, 29:225-228.