title: Tat PTD-endostatin: A novel anti-angiogenesis protein with ocular barrier permeability via eye-drops.
abstract: BACKGROUND
  Endostatin, a specific inhibitor of endothelial cell proliferation and angiogenesis, has been proved to have effects on ocular neovascular diseases by intraocular injection. In order to increase its permeability to ocular barriers and make it effective on fundus oculi angiogenesis diseases via non-invasive administration (eye drops), endostatin was fused to Tat PTD via a genetic engineering method.
  
  
  METHODS
  Most of the Tat PTD- endostatin was expressed as inclusion bodies in Escherichia coli, so pure and active Tat PTD-endostatin was prepared by a series of operations, including inclusion body denaturation, refolding and chromatography. The anti-angiogenesis activity of Tat PTD-endostatin was investigated by cell proliferation experiments and chick embryo chorioallantoic membrane assay. In addition, its translocating ability and concrete entry mechanism into cells were also investigated by fluorescence microscope and flow cytometry. The penetrating ability to ocular barriers was also studied by immunohistochemistry. A mouse choroidal neovascularization model was established to investigate the pharmacodynamics of Tat PTD-endostatin.
  
  
  RESULTS
  The obtained Tat PTD-endostatin had excellent anti-angiogenesis activity and was superior to Es in cellular translocating. Macropinocytosis may be the dominant route of entry of Tat PTD-endostatin into cells. Tat PTD-endostatin could cross ocular barriers and arrive at the retina after eye-drop administration. In addition, it displayed inhibitory effects on choroidal neovascularization via eye drops.
  
  
  CONCLUSIONS
  Tat PTD-endostatin possessed excellent ocular penetrating ability and anti-angiogenesis effects.
  
  
  CONCLUSIONS
  Tat PTD is a promising ocular delivery tool, and Tat PTD-endostatin is a potential drug for curing fundus oculi angiogenesis diseases.
authors: Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan, China.
authors: Xinke
authors: X
authors: Zhang
authors: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Jinan, China.
authors: Yan
authors: Y
authors: Li
authors: Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan, China.
authors: Yanna
authors: Y
authors: Cheng
authors: National Glycoengineering Research Center, Shandong University, Jinan, China.
authors: Haining
authors: H
authors: Tan
authors: Department of Ophthalmology, Provincial Hospital Affiliated to Shandong University, Jinan, China.
authors: Zhiwei
authors: Z
authors: Li
authors: Department of Health Care, QiLu Hospital of Shandong University, Jinan, China.
authors: Yi
authors: Y
authors: Qu
authors: Department of Ophthalmology, Provincial Hospital Affiliated to Shandong University, Jinan, China.
authors: Guoying
authors: G
authors: Mu
authors: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Jinan, China. Electronic address: fswang@sdu.edu.cn.
authors: Fengshan
authors: F
authors: Wang
mesh_terms: D060433:Administration, Ophthalmic; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage* / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply*; D020256:Choroidal Neovascularization / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D043169:Endostatins / Q000008:administration & dosage* / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005123:Eye / Q000098:blood supply* / Q000378:metabolism; D058449:Intravitreal Injections; D008297:Male; D008810:Mice, Inbred C57BL; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009883:Ophthalmic Solutions; D010539:Permeability; D010873:Pinocytosis; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage; D013997:Time Factors
chemical_list: D020533:Angiogenesis Inhibitors; D043169:Endostatins; D009883:Ophthalmic Solutions; D011993:Recombinant Fusion Proteins; C000600678:Tat PTD-endostatin
keywords: CNV; Endostatin; Fusion protein; Ocular barrier; Tat PTD