Preparation, characterization and anti-angiogenesis activity of endostatin covalently modified by polysulfated heparin.
To improve the stability and anti-angiogenesis activity of endostatin (ES), ES was modified by polysulfated heparin (PSH). SDS-PAGE and free amino group determination were employed to study purity and modification procedure. The inhibition of ES and PSH-ES on endothelial cell proliferation, chorioallantoic membrane (CAM) angiogenesis and choroidal neovascularization (CNV) were studied. Western blotting was employed to study the effects on the expression of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF). Changes of the secondary structure were analyzed by circular dichroism (CD) spectra and heat stability was also studied. Our study indicated that the modified product had a better heat tolerance than ES and its anti-angiogenesis activity in CAM model and CNV model were better than that of ES. More obvious down-regulation of VEGF and up-regulation of PEDF effects of PSH-ES than ES in chorioid tissues were detected. The result of CD analysis suggested that little secondary structure change was detected compared with that of ES. Compared with native ES, PSH-ES is a potential anti-tumor drug with better heat stability and better anti-angiogenesis activity both in CAM and CNV models.
School of Pharmaceutical Sciences, Shandong University, Jinan, China, 250012.
Tan Hai
TH
Ning
Cao Ji
CJ
Chao
Mu Guo
MG
Ying
Xiao
X
Min
Wang Feng
WF
Shan
D000596:Amino Acids / Q000737:chemistry; D020533:Angiogenesis Inhibitors / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D002829:Choroid / Q000187:drug effects / Q000378:metabolism; D002942:Circular Dichroism; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D043169:Endostatins / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D005136:Eye Proteins / Q000096:biosynthesis; D006493:Heparin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D006358:Hot Temperature; D007202:Indicators and Reagents; D007834:Lasers; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009414:Nerve Growth Factors / Q000096:biosynthesis; D015843:Serpins / Q000096:biosynthesis; D013778:Tetrazolium Salts; D013844:Thiazoles; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis; D000099174:Pigment Epithelium-Derived Factor
D000596:Amino Acids; D020533:Angiogenesis Inhibitors; D043169:Endostatins; D005136:Eye Proteins; D006493:Heparin; D007202:Indicators and Reagents; D009414:Nerve Growth Factors; D015843:Serpins; D013778:Tetrazolium Salts; D013844:Thiazoles; D042461:Vascular Endothelial Growth Factor A; D000099174:Pigment Epithelium-Derived Factor; C022616:thiazolyl blue