Production and characterization of monoclonal anti-sphingosine-1-phosphate antibodies.
Sphingosine-1-phosphate (S1P) is a pleiotropic bioactive lipid involved in multiple physiological processes. Importantly, dysregulated S1P levels are associated with several pathologies, including cardiovascular and inflammatory diseases and cancer. This report describes the successful production and characterization of a murine monoclonal antibody, LT1002, directed against S1P, using novel immunization and screening methods applied to bioactive lipids. We also report the successful generation of LT1009, the humanized variant of LT1002, for potential clinical use. Both LT1002 and LT1009 have high affinity and specificity for S1P and do not cross-react with structurally related lipids. Using an in vitro bioassay, LT1002 and LT1009 were effective in blocking S1P-mediated release of the pro-angiogenic and prometastatic cytokine, interleukin-8, from human ovarian carcinoma cells, showing that both antibodies can out-compete S1P receptors in binding to S1P. In vivo anti-angiogenic activity of all antibody variants was demonstrated using the murine choroidal neovascularization model. Importantly, intravenous administration of the antibodies showed a marked effect on lymphocyte trafficking. The resulting lead candidate, LT1009, has been formulated for Phase 1 clinical trials in cancer and age-related macular degeneration. The anti-S1P antibody shows promise as a novel, first-in-class therapeutic acting as a "molecular sponge" to selectively deplete S1P from blood and other compartments where pathological S1P levels have been implicated in disease progression or in disorders where immune modulation may be beneficial.
O'Brien
Nicole
N
Lpath Inc., San Diego, CA 92121, USA.
Jones
S Tarran
ST
Williams
David G
DG
Cunningham
H Brad
HB
Moreno
Kelli
K
Visentin
Barbara
B
Gentile
Angela
A
Vekich
John
J
Shestowsky
William
W
Hiraiwa
Masao
M
Matteo
Rosalia
R
Cavalli
Amy
A
Grotjahn
Douglas
D
Grant
Maria
M
Hansen
Geneviève
G
Campbell
Mary-Ann
MA
Sabbadini
Roger
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000494:pharmacology; D000918:Antibody Specificity / Q000276:immunology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D020256:Choroidal Neovascularization / Q000517:prevention & control; D003429:Cross Reactions / Q000276:immunology; D005260:Female; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism; D007700:Kinetics; D008246:Lysophospholipids / Q000276:immunology* / Q000378:metabolism; D008268:Macular Degeneration / Q000188:drug therapy; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D016296:Mutagenesis; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology; D011485:Protein Binding; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007378:Interleukins; D008246:Lysophospholipids; C060506:sphingosine 1-phosphate; D013110:Sphingosine