Recombinant anti-vascular endothelial growth factor fusion protein efficiently suppresses choridal neovasularization in monkeys.
OBJECTIVE
KH902 is a fusion protein which combines ligand binding elements taken from the extracellular domains of vascular endothelial growth factor (VEGF) receptors 1 and 2 and the Fc portion of IgG1. This study is designed to examine the inhibitory effect of KH902 in the choroidal neovascularization (CNV) monkey model.


METHODS
The binding affinity with VEGF was measured by using the human VEGF ELISA kit, and the biological activity effect of KH902 was assayed by an in vitro inhibition experiment on human umbilical vein endothelial cell proliferation that was induced by VEGF. The experimental CNV was induced by causing perimacular laser injury in the eyes of rhesus monkeys and confirmed by fluorescence fundus angiography (FFA), optical coherence tomography (OCT), and multifocal electroretinograms (mf-ERG) 20 days after the infliction of the laser injury. KH902 was delivered to the animals through intravitreal injection at various doses. Monkeys were observed four weeks after injection by ophthalmic examination, FFA, OCT, mf-ERG, histopathology, and immunohistochemistry analysis.


RESULTS
KH902 binds VEGF at a high affinity with a mean of IC(50) of 10 pM. KH902 at 41 nM can completely block VEGF-induced cell proliferation and KH902 at 10.7 nM can block 82.6% of cell growth. In the eyes of the treatment group, which received 300 microg and 500 microg KH902, choroidal neovascularization leakage was obviously less than before injection, and no leakage was observed at the end of the observation after injection. No high reflect light echogenic mass was detected by OCT. However, in the 0.1 mg KH902-treated and control eyes, the leakage and high reflect light echogenic mass still existed. The reduction of experimental CNV was greater in eyes treated with 300 microg and 500 microg KH902 than in eyes treated with 0.1 mg KH902 and the control eyes. There were fiber-vasculosa membrane proliferation in the 100 microg KH902-treated eyes and control eyes but not in the 300 microg and 500 microg KH902-treated eyes under histopathologic observation. The results of mf-ERG demonstrated that there was greater improvement in the 300 microg and 500 microg KH902-treated eyes than in the 100 microg KH902-treated eyes and control eyes.


CONCLUSIONS
KH902 presents high affinity with VEGF and inhibitory activity on the proliferation of human umbilical vein endothelial cells (HUVECs) induced by VEGF. A single 300 microg or 500 microg KH902 intravitreal injection effectively inhibited leakage and growth of the CNV in rhesus monkeys without evidence of toxicity. This study suggests that KH902 has promise as a local antiangiogenic treatment of CNV.
Zhang
Ming
M
Department of Ophthalmology, West China Hospital, Sichuan University, China.
Zhang
Junjun
J
Yan
Mi
M
Li
Hong
H
Yang
Chun
C
Yu
Dechao
D
D000818:Animals; D001667:Binding, Competitive; D002199:Capillary Permeability / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D020256:Choroidal Neovascularization / Q000175:diagnosis / Q000473:pathology* / Q000503:physiopathology*; D004596:Electroretinography; D042783:Endothelial Cells / Q000166:cytology; D005451:Fluorescein Angiography; D005654:Fundus Oculi; D006801:Humans; D007267:Injections; D008253:Macaca mulatta; D008266:Macula Lutea / Q000503:physiopathology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D041623:Tomography, Optical Coherence; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism / Q000494:pharmacology; D014822:Vitreous Body
D011993:Recombinant Fusion Proteins; D042461:Vascular Endothelial Growth Factor A; C527363:KH902 fusion protein