Research on the ocular safety of the developed pirfenidone eye drops application

doi:10.3877/cma.j.issn.2095-2007.2017.06.003

Abstract

Abstract:

Objective

To study the ocular safety of the developed pirfenidone(PFD) eye drops application.

Methods

18 isolated rats were selected. The rats were divided into the control group, the experiment 1 groups and the experiment 2 groups according to the random number table method, with 6 rats in each group. The rats in the control group were treated with phosphate buffer (PBS). The 1 groups and the experimental 2 groups were eyedrops of 3 mg/ml and 30 mg/ml PFD respectively. All eyes were dropped 6 times a day for 14 days.The irritant behavior of the rats was observed and recorded after each eye drops. It includes the eye, frequent blinking or head scratching. The conjunctival, corneal and iris tissues were observed under slit lamp microscope. Fluorescein sodium staining was used to check whether there was any damage to the cornea. According to Draize′s rabbit eye irritation test score, rats were scored. The damage of the ocular surface of the three groups of rats was compared for fourteenth days.At the end of the three group of rats, fourteenth days after the experiment, 10% chloral chloral aldehyde was assigned to 3.5 ml/kg, and the rats were injected intraperitoneally. After anesthesia, the conjunctiva and cornea of rats were taken under the microscope of eye surgery. They were placed in 4% neutral buffer formaldehyde fixative for 4 hours. Pathological hematoxylin eosin staining was performed, and observed under microscope.The number and percentage of the stimuli were described by the χ² test.The duration of stimulation behavior and the score of Draize eye irritation test were described with mean standard deviation (±s), and the single factor analysis of variance was used.

Results

The control group, experimental group 1 and experimental group 2 rats in the use of eye drops, eye immediately scratching behavior, disappeared last about 20~30 minutes. The statistical results showed that the rats in the control group, the experiment group 1 and the experimental group 2 were treated with eye drops, there was no significant difference in stimulus behavior, and there was no statistical difference (χ²=-1.098, P>0.05). The duration of stimulation in the control group, the group1 and the group 2 were (23.50±1.87)min, (24.83±3.06)min and (25.67±2.81)min. The difference was not statistically significant (F=1.04, P>0.05). The control group, experimental group 1 and experimental group 2 of the conjunctiva, cornea and iris Draize score were 0.33±0.52, 0.50±0.55 and 0.83±0.75, way analysis of variance results showed that the control group, experimental group 1 and experimental group 2 of the conjunctiva, cornea and iris Draize score difference was not statistically significant (F=1.03, P>0.05). Hematoxylin-eosin staining demonstrated that the morphology and structure of ocular tissues in the experimental group one, the experimental group two and control group were normal, and there were no significant pathological changes.

Conclusion

The local application of PFD eye drops and the safety of cornea are good and have good prospects for application.

Key words: Pirfenidone eye drops, Cornea, Safety

Gongfa Wu, Lizhen Qiu, Yuting Zeng, Qiting Huang, Yujun Liu, Haigang Li. Research on the ocular safety of the developed pirfenidone eye drops application[J]. Chinese Journal of Ophthalmologic Medicine(Electronic Edition), 2017, 07(06): 250-256.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhykyxzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-2007.2017.06.003

https://zhykyxzz.cma-cmc.com.cn/EN/Y2017/V07/I06/250

Figures/Tables 4

References 67

[1]	黄晓波,邹海东,王宁, 等. 上海市北新泾街道60岁及以上人群盲和视力损伤的现况调查[J]. 中华眼科杂志, 2009, 45(9): 786-792.
[2]	李运,毕宏生,王利华, 等. 山东省农村50岁及以上人群盲和中、重度视力损伤病因分析[J]. 中华眼科杂志, 2013, 49(2): 144-150.
[3]	尉洋,向云,生侠, 等. 海口市郊≥50岁人群的视力损伤及患盲病率调查[J]. 国际眼科杂志, 2012, 12(9): 1806-1808.
[4]	Whitcher JP,Srinivasan M,Upadhyay MP. Corneal blindness: a global perspective[J]. Bull World Health Organ, 2001, 79(3): 214-221.
[5]	George PM,Wells AU. Pirfenidone for the treatment of idiopathic pulmonary fibrosis[J]. Expert Rev Clin Pharmacol, 2017, 10(5): 483-491.
[6]	国家科学技术委员会. 实验动物管理条例[Z]. 2013－7－18.
[7]	Draize JH,Woodard G,Calvery HO. Methods for the Study of Irritation and Toxicity of Substances Applied Topically to the Skin and Mucous Membranes[J]. J Pharmacol Expe Ther, 1944, 6(3): 377-390.
[8]	Meek KM,Knupp C. Corneal structure and transparency[J]. Prog Retin Eye Res, 2015, 49(2): 1-16.
[9]	黄一飞,王大江,王丽强, 等. 人工角膜在治疗严重化学烧伤眼中的研究[J]. 中华眼科杂志, 2007, 43(4): 297-302.
[10]	Nili－Ahmadabadi M,Oskouee SJ,Hashemian MN, et al. Epidemiology of ocular trauma in Tehran, Iran: A retrospective survey of 959 cases[J]. Int J Ophthalmol, 2008, 8(10): 1975-1979.
[11]	MGG Jr,Xie A,Owsley C. Rate of eye injury in the United States[J]. Arch Ophthalmol, 2005, 123(7): 970-976.
[12]	Hilber D,Mitchener TA,Stout J, et al. Eye injury surveillance in the U. S. Department of Defense, 1996－2005[J]. Am J Prev Med, 2010, 38(Suppl. 1): 78-85.
[13]	MGG Jr,Owsley C. Incidence of emergency department－treated eye injury in the United States[J]. Arch Ophthalmol, 2005, 123(5): 662-666.
[14]	West－Mays JA,Dwivedi DJ. The keratocyte: corneal stromal cell with variable repair phenotypes[J]. Int J Biochem Cell Biol, 2006, 38(10): 1625-1631.
[15]	樊廷俊,白苏冉. 角膜基质创伤愈合的研究进展[J]. 山东大学学报(理学版), 2016, 51(3): 1-10.
[16]	Hassell JR,Birk DE. The molecular basis of corneal transparency[J]. Exp Eye Res, 2010, 91(3): 326-333.
[17]	许杨,肖骏,杨晓梅, 等. 准分子激光治疗12例角膜瘢痕的效果观察[J]. 浙江预防医学, 2000, 12(6): 50-51.
[18]	高桂平,邵毅,周琼, 等. 260例角膜移植常见病因分析[J]. 江西医药, 2007, 42(1): 50-51.
[19]	李小磊,宋秀君,卢建民, 等. 姜黄素对角膜基质细胞纤维化的抑制作用[J]. 中华实验眼科杂志, 2011, 29(5): 402-406.
[20]	牟章兵,杜之渝,晏丕松, 等. 汉防己甲素等5种抗纤维化药物对离体兔角膜基质细胞的抑制作用[J]. 眼科新进展, 2006, 26(8): 592-596.
[21]	李红,张作明,杨新光, 等. 退翳眼液对兔角膜瘢痕厚度及面积影响的实验研究[J]. 中国临床康复, 2004, 8(11): 2091-2093.
[22]	Ke Y,Wu Y,Cui X, et al. Polysaccharide hydrogel combined with mesenchymal stem cells promotes the healing of corneal alkali burn in rats[J]. PLoS One, 2015, 10(3): e0119725.
[23]	Giulio F,Fabio B,Chiara G, et al. Safety and efficacy of topical infliximab in a mouse model of ocular surface scarring[J]. Invest Ophthalmol Vis Sci, 2013, 54(3): 1680-1688.
[24]	Stramer BM,Zieske JD,Jung JC, et al. Molecular mechanisms controlling the fibrotic repair phenotype in cornea: implications for surgical outcomes[J]. Invest Ophthalmol Vis Sci, 2003, 44(10): 4237-4246.
[25]	Milani BY,Milani FY,Park DW, et al. Rapamycin inhibits the production of myofibroblasts and reduces corneal scarring after photorefractive keratectomy[J]. Invest Ophthalmol Vis Sci, 2013, 54(12): 7424-7430.
[26]	唐敏,席头华,唐罗生, 等. 人羊膜匀浆提取液对兔角膜成纤维细胞增殖和TGF－β1 mRNA表达的影响[J]. 眼科新进展, 2013, 33(11): 1029-1032.
[27]	Owens GM. Strategies to manage costs in idiopathic pulmonary fibrosis[J]. Am J Manag Care, 2017, 23(Suppl. 11): 191-196.
[28]	Tzouvelekis A,Bouros E,Tzilas V, et al. Pirfenidone in Idiopathic Pulmonary Fibrosis "RECAP－itulating Safety into the Real World"[J]. Respiration, 2017, 94(5): 405-407.
[29]	徐建华,肖红波,程玉花, 等. 吡非尼酮对大鼠肾间质纤维化的防治作用研究[J]. 中国现代医学杂志, 2017, 27(19): 1-6.
[30]	肖敏,屈小虎,吕聚坪, 等. 吡非尼酮对四氯化碳诱导的小鼠肝纤维化的影响[J]. 中国应用生理学杂志, 2016, 32(4): 378-382.
[31]	Iyer SN,Wild JS,Schiedt MJ, et al. Dietary intake of pirfenidone ameliorates bleomycin－induced lung fibrosis in hamsters[J]. Journal of Laboratory & Clinical Medicine, 1995, 125(6): 779-785.
[32]	Tzouvelekis A,Ntolios P,Karampitsakos T, et al. Safety and efficacy of pirfenidone in severe Idiopathic Pulmonary Fibrosis a real－world observational study[J]. Pulm Pharmacol Ther, 2017, 46(5): 48-53.
[33]	王红梅,杨佳丹,蒙龙, 等. 吡非尼酮治疗特发性肺纤维化疗效与安全性的系统评价[J]. 中国药房, 2016, 27(3): 345-348.
[34]	付麟紫,吴晓梅. 特发性肺纤维化药物治疗推荐[J]. 中华老年多器官疾病杂志, 2017, 16(3): 234-237.
[35]	李云霞,李琼,加慧, 等. 吡非尼酮治疗特发性肺纤维化的临床研究[J]. 航空航天医学杂志, 2016, 27(11): 1365-1367.
[36]	陈钰清,闫永吉,仇美华, 等. 吡非尼酮治疗肺纤维化的疗效与安全性meta分析[J]. 临床药物治疗杂志, 2016, 14(4): 13-22.
[37]	陈俊杰,吴共发,林俊汕, 等. 吡非尼酮对大鼠角膜基质细胞增殖的影响[J]. 国际眼科杂志, 2015, 15 (2): 201-204.
[38]	Liu H,Drew P,Gaugler AC, et al. Pirfenidone inhibits lung allograft fibrosis through L－arginine－arginase pathway[J]. American Journal of Transplantation, 2015, 5(6): 1256-1263.
[39]	Choi K,Lee K,Ryu SW, et al. Pirfenidone inhibits transforming growth factor－β1－induced fibrogenesis by blocking nuclear translocation of Smads in human retinal pigment epithelial cell line ARPE－19[J]. Molecular Vision, 2012, 18(104): 1010-1020.
[40]	Iyer SN,Gurujeyalakshmi G,Giri SN. Effects of Pirfenidone on Procollagen Gene Expression at the Transcriptional Level in Bleomycin Hamster Model of Lung Fibrosis[J]. Journal of Pharmacology & Experimental Therapeutics, 1999, 289(1): 211-218.
[41]	Hewitson TD,Kelynack KJ,Tait MG, et al. Pirfenidone reduces in vitro rat renal fibroblast activation and mitogenesis[J]. Journal of Nephrology, 2001, 14(6): 453-460.
[42]	Gurujeyalakshmi G,Hollinger MA,Giri SN. Pirfenidone inhibits PDGF isoforms in bleomycin hamster model of lung fibrosis at the translational level[J]. American Journal of Physiology, 1999, 276(1): 311-318.
[43]	Oku H,Nakazato H,Horikawa T, et al. Pirfenidone suppresses tumor necrosis factor－alpha, enhances interleukin－10 and protects mice from endotoxic shock[J]. European Journal of Pharmacology, 2002, 446(1): 167-176.
[44]	Nakazato H,Oku H,Yamane S, et al. A novel anti－fibrotic agent pirfenidone suppresses tumor necrosis factor－alpha at the translational level[J]. European Journal of Pharmacology, 2002, 446(1): 177-185.
[45]	Oku H,Shimizu T,Kawabata T, et al. Antifibrotic action of pirfenidone and prednisolone: different effects on pulmonary cytokines and growth factors in bleomycin－induced murine pulmonary fibrosis[J]. European Journal of Pharmacology, 2008, 590(1): 400-408.
[46]	Grattendick KJ,Nakashima JM,Feng L, et al. Effects of three anti－TNF－alpha drugs: etanercept, infliximab and pirfenidone on release of TNF－alpha in medium and TNF－alpha associated with the cell in vitro[J]. Int Immunopharmacol, 2008, 8(5): 679-687.
[47]	Schaefer CJ,Ruhrmund DW,Pan L, et al. Antifibrotic activities of pirfenidone in animal models[J]. Eur Respir Rev, 2011, 20(120): 85-97.
[48]	杜磊,王昌源,张立华, 等. 吡非尼酮对实验性大鼠肝纤维化的作用[J]. 山东大学学报(医学版), 2009, 47(11): 50-54.
[49]	王琳娜,陈常勇,郭存霞, 等. 吡非尼酮对转化生长因子－β1诱导的大鼠心肌成纤维细胞增殖和胶原蛋白合成的影响[J]. 中国现代医学杂志, 2015, 25(29): 30-34.
[50]	姬巍,蒋恒,杨伟志, 等. 吡非尼酮防治放射性肺纤维化实验研究[J]. 中华放射肿瘤学杂志, 2010, 19(6): 560-563.
[51]	Tian XL,Yao W,Guo ZJ, et al. Low dose pirfenidone suppresses transforming growth factor beta－1 and tissue inhibitor of metalloproteinase－1, and protects rats from lung fibrosis induced by bleomycina[J]. Chin Med Sci J, 2006, 21(3): 145-151.
[52]	Waller JR,Murphy GJ,Bicknell GR, et al. Pirfenidone inhibits early myointimal proliferation but has no effect on late lesion size in rats[J]. Eur J Vasc Endovasc Surg, 2002, 23(3): 234-240.
[53]	Lin X,Yu M,Wu K, et al. Effects of pirfenidone on proliferation, migration, and collagen contraction of human Tenon′s fibroblasts in vitro[J]. Invest Ophthalmol Vis Sci, 2009, 50(8): 3763-3770.
[54]	Zhong H,Sun G,Lin X, et al. Evaluation of pirfenidone as a new postoperative anti－scarring agent in experimental glaucoma surgery[J]. Invest Ophthalmol Vis Sci, 2011, 52(6): 3136-3142.
[55]	Yang Y,Ye Y,Lin X, et al. Inhibition of Pirfenidone on TGF－beta2 Induced Proliferation, Migration and Epithlial－Mesenchymal Transition of Human Lens Epithelial Cells Line SRA01/04[J]. Plos One, 2013, 8(2): e56837.
[56]	Kim H,Choi YH,Park SJ, et al. Antifibrotic effect of Pirfenidone on orbital fibroblasts of patients with thyroid－associated ophthalmopathy by decreasing TIMP－1 and collagen levels[J]. Invest Ophthalmol Vis Sci, 2010, 51(6): 3061-3066.
[57]	Chung SA,Jeon BK,Choi YH, et al. Pirfenidone attenuates the IL－1β－induced hyaluronic acid increase in orbital fibroblasts from patients with thyroid－associated ophthalmopathy[J]. Invest Ophthalmol Vis Sci, 2014, 55(4): 2276-2283.
[58]	李亨辉,李云燕,邵毅, 等. 吡非尼酮滴眼液在兔眼青光眼手术中应用研究[J]. 中国实用眼科杂志, 2015, 33(10): 1179-1184.
[59]	Wilson SE. Corneal myofibroblast biology and pathobiology: Generation, persistence, and transparency[J]. Experimental Eye Research, 2012, 99(1): 78-88.
[60]	邵毅,余瑶,裴重刚, 等. 吡非尼酮滴眼液在兔眼小梁切除术后抗瘢痕作用研究[J]. 中国现代医学杂志, 2015, 25(2): 13-17.
[61]	Nakayama S,Mukae H,Sakamoto N, et al. Pirfenidone inhibits the expression of HSP47 in TGF－beta1－stimulated human lung fibroblasts.[J]. Life Sciences, 2008, 82(3): 210-217.
[62]	Suga H,Teraoka S,Ota K, et al. Preventive effect of pirfenidone against experimental sclerosing peritonitis in rats[J]. Experimental & Toxicologic Pathology, 1995, 47(4): 287-291.
[63]	Garcã AL,Hernã NL,Sandoval A, et al. Pirfenidone effectively reverses experimental liver fibrosis[J]. Journal of Hepatology, 2002, 37(6): 797-805.
[64]	Ramachandrarao SP,Zhu Y,Ravasi T, et al. Pirfenidone Is Renoprotective in Diabetic Kidney Disease[J]. Journal of the American Society of Nephrology Jasn, 2009, 20(8): 1249-1256.
[65]	Hijikawa T,Kwon AH. Effects of pirfenidone on endotoxin－induced liver injury after partial hepatectomy in rats[J]. Transplant Proc, 2004, 36(7): 1975-1976.
[66]	Gurujeyalakshmi G,Hollinger MA,Giri SN. Pirfenidone inhibits PDGF isoforms in bleomycin hamster model of lung fibrosis at the translational level[J]. American Journal of Physiology, 1999, 276(2 Pt 1): 311-318.
[67]	Nagai S,Hamada K,Shigematsu M, et al. Open－label compassionate use one year－treatment with pirfenidone to patients with chronic pulmonary fibrosis[J]. Internal Medicine, 2002, 41(12): 1118-1123.

组织	损伤情况		分值
角膜	?	A.混浊(以最致密部位计算)	?
	散在或弥漫性混浊，虹膜清晰可见		1
	半透明区易分辨，虹膜清晰可见		2
	出现乳白色区域，虹膜细节不清，瞳孔大小勉强可见		3
	角膜不透明，虹膜无法辨认		4
	?	B.角膜损害面积	?
	?	≤25%	1
	?	>25%，≤50%	2
	?	>50%，≤75%	3
	?	>75%	4
	?	积分＝A×B×5	?
虹膜	?	A.数值	?
	皱褶加深、充血及肿胀，角膜周围轻度充血，瞳孔对光有反应		1
	?	出血、肉眼可见坏死，对光无反应	2
	?	积分＝A×5	?
结膜	?	A.睑结膜充血	?
	充血重于正常血管		1
	呈深红色弥漫性充血，血管不易分辨		2
	呈紫红色充血		3
	?	B.水肿	?
	任何重于正常的肿胀		1
	明显水肿，伴部分眼睑外翻		2
	水肿至眼睑闭合近一半		3
	水肿至眼睑闭合超过一半		4
	?	C.分泌物	?
	分泌物异于正常		1
	分泌物仅使眼睑和睫毛潮湿		2
	分泌物使眼睑和眼周大面积潮湿		3
	?	积分＝(A＋B＋C)×2	?

组织	损伤情况		分值
角膜	?	A.混浊(以最致密部位计算)	?
	散在或弥漫性混浊，虹膜清晰可见		1
	半透明区易分辨，虹膜清晰可见		2
	出现乳白色区域，虹膜细节不清，瞳孔大小勉强可见		3
	角膜不透明，虹膜无法辨认		4
	?	B.角膜损害面积	?
	?	≤25%	1
	?	>25%，≤50%	2
	?	>50%，≤75%	3
	?	>75%	4
	?	积分＝A×B×5	?
虹膜	?	A.数值	?
	皱褶加深、充血及肿胀，角膜周围轻度充血，瞳孔对光有反应		1
	?	出血、肉眼可见坏死，对光无反应	2
	?	积分＝A×5	?
结膜	?	A.睑结膜充血	?
	充血重于正常血管		1
	呈深红色弥漫性充血，血管不易分辨		2
	呈紫红色充血		3
	?	B.水肿	?
	任何重于正常的肿胀		1
	明显水肿，伴部分眼睑外翻		2
	水肿至眼睑闭合近一半		3
	水肿至眼睑闭合超过一半		4
	?	C.分泌物	?
	分泌物异于正常		1
	分泌物仅使眼睑和睫毛潮湿		2
	分泌物使眼睑和眼周大面积潮湿		3
	?	积分＝(A＋B＋C)×2	?

分组	刺激性行为			持续时间(min)
分组	－[%(例数/总例数)]	＋[%(例数/总例数)]	＋＋[%(例数/总例数)]	持续时间(min)
对照组	0.00(0/6)	83.33(5/6)	16.67(1/6)	23.50±1.87
实验1组	0.00(0/6)	50.00(3/6)	50.00(3/6)	24.83±3.06
实验2组	0.00(0/6)	50.00(3/6)	50.00(3/6)	25.67±2.81
统计量	χ²＝1.77			F＝1.04
P值	>0.05			>0.05

分组	刺激性行为			持续时间(min)
分组	－[%(例数/总例数)]	＋[%(例数/总例数)]	＋＋[%(例数/总例数)]	持续时间(min)
对照组	0.00(0/6)	83.33(5/6)	16.67(1/6)	23.50±1.87
实验1组	0.00(0/6)	50.00(3/6)	50.00(3/6)	24.83±3.06
实验2组	0.00(0/6)	50.00(3/6)	50.00(3/6)	25.67±2.81
统计量	χ²＝1.77			F＝1.04
P值	>0.05			>0.05

Please choose a citation manager

Content to export