title: Tat PTD-Endostatin-RGD: A novel protein with anti-angiogenesis effect in retina via eye drops.
abstract: Diabetic retinopathy is a leading cause of blindness. The objective was to design a novel fusion protein, Tat PTD-Endostatin-RGD, to treat retinal neovascularization via eye drops instead of traditional intravitreal injection trepapeutical methods.
  
  
  
  The anti-angiogenesis ability was evaluated in vitro by chick embryo chorioallantoic membrane assay, wound healing assay and tube formation assay. Corneal barrier and blood-retina barrier were constructed in vitro to investigate the penetration ability of Tat PTD-Endostatin-RGD. Western blot was used to detect the integrin αvβ3 expression level in rat retina microvascular endothelial cells which was stimulated by S-nitroso-N-acetylpenicillamine. The binding affinity of Tat PTD-Endostatin-RGD to integrin αvβ3 was investigated by evaluating the penetration ability on blood-retina barriers treated with S-nitroso-N-acetylpenicillamine. The pharmacodynamics and efficacy analysis were further carried out in the oxygen-induced retinopathy model in vivo. In addition, the pharmacokinetic profile via eye drops was studied on a C57BL/6 mice model.
  
  
  
  Tat PTD-Endostatin-RGD showed high anti-angiogenesis activity and high ability to penetrate these two barriers in vitro. The Western blot results indicated S-nitroso-N-acetylpenicillamine upregulated the expression level of integrin αvβ3 in a dose-dependent manner. Tat PTD-Endostatin-RGD showed a high affinity to rat retina microvascular endothelial cells treated with S-nitroso-N-acetylpenicillamine. The results showed that Tat PTD-Endostatin-RGD could inhibit abnormal angiogenesis in retina via eye drops.
  
  
  
  Tat PTD-Endostatin-RGD showed high penetration ability through ocular barriers, bound specifically to integrin αvβ3 and effectively inhibited the abnormal angiogenesis.
  
  
  
  Tat PTD-Endostatin-RGD represents a potent novel drug applied via eye drops for fundus oculi neovascularization diseases.
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Lian
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Ophthalmology, Provincial Hospital Affiliated to Shandong University, Jinan 250012, China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Juzheng
authors.initials: J
authors.lastname: Sheng
authors.affiliation: Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Xinke
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Danyang
authors.initials: D
authors.lastname: Feng
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Fengxin
authors.initials: F
authors.lastname: Yin
authors.affiliation: Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
authors.forename: Aijun
authors.initials: A
authors.lastname: Wang
authors.affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China; National Glycoengineering Research Center, Shandong University, Jinan 250012, China. Electronic address: fswang@sdu.edu.cn.
authors.forename: Fengshan
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D001813:Blood-Retinal Barrier / Q000187:drug effects; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000187:drug effects; D003930:Diabetic Retinopathy / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D043169:Endostatins / Q000008:administration & dosage* / Q000378:metabolism; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D009842:Oligopeptides / Q000008:administration & dosage* / Q000378:metabolism; D009883:Ophthalmic Solutions / Q000008:administration & dosage; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000378:metabolism; D012160:Retina / Q000187:drug effects / Q000473:pathology; D015861:Retinal Neovascularization / Q000188:drug therapy* / Q000473:pathology; D012171:Retinal Vessels / Q000187:drug effects / Q000473:pathology; D026423:S-Nitroso-N-Acetylpenicillamine / Q000378:metabolism
chemical_list: D043169:Endostatins; D039302:Integrin alphaVbeta3; D009842:Oligopeptides; D009883:Ophthalmic Solutions; D011993:Recombinant Fusion Proteins; C000600678:Tat PTD-endostatin; C047981:arginyl-glycyl-aspartic acid; D026423:S-Nitroso-N-Acetylpenicillamine
keywords: Blood-retina barrier; Corneal barriers; Neovascularization; Oxygen-induced retinopathy