Antiangiogenic activity of semisynthetic biotechnological heparins: low-molecular-weight-sulfated Escherichia coli K5 polysaccharide derivatives as fibroblast growth factor antagonists.
OBJECTIVE
Low-molecular-weight heparin (LMWH) exerts antitumor activity in clinical trials. The K5 polysaccharide from Escherichia coli has the same structure as the heparin precursor. Chemical and enzymatic modifications of K5 polysaccharide lead to the production of biotechnological heparin-like compounds. We investigated the fibroblast growth factor-2 (FGF2) antagonist and antiangiogenic activity of a series of LMW N,O-sulfated K5 derivatives.


RESULTS
Surface plasmon resonance analysis showed that LMW-K5 derivatives bind FGF2, thus inhibiting its interaction with heparin immobilized to a BIAcore sensor chip. Interaction of FGF2 with tyrosine-kinase receptors (FGFRs), heparan sulfate proteoglycans (HSPGs), and alpha(v)beta3 integrin is required for biological response in endothelial cells. Similar to LMWH, LMW-K5 derivatives abrogate the formation of HSPG/FGF2/FGFR ternary complexes by preventing FGF2-mediated attachment of FGFR1-overexpressing cells to HSPG-bearing cells and inhibit FGF2-mediated endothelial cell proliferation. However, LMW-K5 derivatives, but not LMWH, also inhibit FGF2/alpha(v)beta3 integrin interaction and consequent FGF2-mediated endothelial cell sprouting in vitro and angiogenesis in vivo in the chick embryo chorioallantoic membrane.


CONCLUSIONS
LMW N,O-sulfated K5 derivatives affect both HSPG/FGF2/FGFR and FGF2/alpha(v)beta3 interactions and are endowed with FGF2 antagonist and antiangiogenic activity. These compounds may provide the basis for the design of novel LMW heparin-like angiostatic compounds.
Presta
Marco
M
Unit of General Pathology and Immunology, Department of Biomedical Sciences and Biotechnology, School of Medicine, University of Brescia, Italy. presta@med.unibs.it
Oreste
Pasqua
P
Zoppetti
Giorgio
G
Belleri
Mirella
M
Tanghetti
Elena
E
Leali
Daria
D
Urbinati
Chiara
C
Bugatti
Antonella
A
Ronca
Roberto
R
Nicoli
Stefania
S
Moroni
Emanuela
E
Stabile
Helena
H
Camozzi
Maura
M
Hernandez
German Andrés
GA
Mitola
Stefania
S
Dell'Era
Patrizia
P
Rusnati
Marco
M
Ribatti
Domenico
D
D020533:Angiogenesis Inhibitors / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D016667:Bacterial Capsules; D016466:CHO Cells / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D042783:Endothelial Cells / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000031:analogs & derivatives / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods*; D019812:Heparan Sulfate Proteoglycans / Q000031:analogs & derivatives / Q000172:deficiency / Q000378:metabolism; D006495:Heparin, Low-Molecular-Weight / Q000096:biosynthesis* / Q000138:chemical synthesis / Q000235:genetics; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis* / Q000235:genetics
D020533:Angiogenesis Inhibitors; D019812:Heparan Sulfate Proteoglycans; D006495:Heparin, Low-Molecular-Weight; D039302:Integrin alphaVbeta3; D011135:Polysaccharides, Bacterial; D016222:Fibroblast Growth Factor 2; C050673:capsular polysaccharide K5; D005346:Fibroblast Growth Factors