Preclinical evaluation of an anti-alpha5beta1 integrin antibody as a novel anti-angiogenic agent.
Integrin alpha5beta1, the principal fibronectin receptor, is an important survival factor, playing a key role in angiogenesis. Angiogenesis is critical for tumor growth, and anti-angiogenic therapies have met clinical success. To validate the therapeutic potential of an anti-alpha5beta1 strategy, we generated volociximab (M200) a chimeric human IgG4 version of the alpha5beta1 function-blocking murine antibody IIA1; and F200, the Fab derivative. Volociximab, F200 and IIA1 showed similar activity by ELISA (EC50= 0.2nM), Biacore (Kd= 0.1-0.4nM) and inhibition of fibronectin binding (IC50= 2-3nM). The inhibitory potential of alpha5beta1 antibodies was compared to HuMV833, an anti-VEGF antibody. Both volociximab and HuMV833 inhibited HUVEC proliferation (IC50 of volociximab = 0.2-0.5nM; IC50 of HuMV833 = 45nM). However, IIA1, volociximab and F200 were also potent inhibitors of an in vitro model of angiogenesis (HUVEC tube formation assay), unlike HuMV833. Additionally, volociximab inhibited in vitro tube formation induced by VEGF and/or bFGF, suggesting a mechanism of action independent of growth factor stimulus. In fact, inhibition of alpha5beta1 function by volociximab induced apoptosis of actively proliferating, but not resting, endothelial cells. Volociximab does not cross-react with rodent alpha5beta1, therefore in vivo validation of an anti-alpha5beta1 approach was conducted in a cynomolgus model of choroidal revascularization. Volociximab and F200 were potent inhibitors of neovessel formation in this model. These data demonstrate that volociximab has therapeutic potential in diseases in which new vessel formation is a component of the pathology.
Ramakrishnan
Vanitha
V
PDL Biopharma Inc, Fremont CA 94555, USA. vanitha@pdl.com
Bhaskar
Vinay
V
Law
Debbie A
DA
Wong
Melanie H L
MH
DuBridge
Robert B
RB
Breinberg
Danna
D
O'Hara
Christopher
C
Powers
David B
DB
Liu
Gao
G
Grove
Jennifer
J
Hevezi
Peter
P
Cass
Kellie M
KM
Watson
Susan
S
Evangelista
Ferdinand
F
Powers
Rick A
RA
Finck
Barbara
B
Wills
Margaret
M
Caras
Ingrid
I
Fang
Yuni
Y
McDonald
Donald
D
Johnson
Dale
D
Murray
Richard
R
Jeffry
Ursula
U
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D000906:Antibodies / Q000627:therapeutic use*; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D058846:Antibodies, Monoclonal, Murine-Derived; D019556:COS Cells; D002522:Chlorocebus aethiops; D004353:Drug Evaluation, Preclinical; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D039081:Integrin alpha5beta1 / Q000737:chemistry / Q000276:immunology*; D007700:Kinetics; D008252:Macaca fascicularis; D008268:Macular Degeneration / Q000188:drug therapy; D009389:Neovascularization, Pathologic; D000069283:Rituximab
D020533:Angiogenesis Inhibitors; D000906:Antibodies; D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D039081:Integrin alpha5beta1; C516631:volociximab; D000069283:Rituximab