Polypeptides, antibody variable domains and antagonists

Abstract

The invention relates to anti-VEGF polypeptides and antibody single variable domains (dAbs) that are resistant to degradation by a protease, as well as antagonists comprising these. The polypeptides, dAbs and antagonists are useful for pulmonary administration, oral administration, delivery to the lung and delivery to the GI tract of a patient, as well as for treating cancer and inflammatory disease, such as arthritis.

Description

[0001]The present invention relates to protease resistant polypeptides, immunoglobulin (antibody) single variable domains and vascular endothelial growth factor (VEGF) antagonists comprising these. The invention further relates to uses, formulations, compositions and devices comprising such anti-VEGF ligands.BACKGROUND OF THE INVENTION[0002]Polypeptides and peptides have become increasingly important agents in a variety of applications, including industrial applications and use as medical, therapeutic and diagnostic agents. However, in certain physiological states, such as Cancer and inflammatory states (e.g., COPD), the amount of proteases present in a tissue, organ or animal (e.g., in the lung, in or adjacent to a tumor) can increase. This increase in proteases can result in accelerated degradation and inactivation of endogenous proteins and of therapeutic peptides, polypeptides and proteins that are administered to treat disease. Accordingly, some agents that have potential for i...

AI Technical Summary

Benefits of technology

[0574]The receptor binding assay described above was used to assess the potency of the DOM15-26-593 dAb-Fc fusion (FIG. 48). It was found that the DOM15-26-593 dAb has enhanced potency in this assay, which establishes the ability of the dAb to block the binding of VEGF to VEGFR2 in vitro. The potency of the DMS1529 was also demonstrated in a HUVEC (Human Umbilical Vein Endothelial Cell) assay, where the ability of VEGF antagonists to block the VEGF stimulated proliferation of HUVE cells is measured. Briefly, approximately 4e3 HUVE cells are dispensed into the wells of a 96-well plate to which is added a mixture of VEGF and a dilution series of the test substance, such that the final concentration of VEGF is 5 ng / ml, or as otherwise determined by a dose-response titration. The cells are incubated at 37° C. for a further 4 days, at which point the cell number is determined by the use of a cell quantification reagent such as “CellTiter”. This allows the colorimetric determination of cell proliferation in comparison with standards over the 4 days of the experiment. Cell numbers are determined at the end of a fixed incubation period with a pre-determined amount of VEGF and a varying amount of test article. The more potent the antagonist, the lower the cell proliferation observed (FIG. 49).

Problems solved by technology

Accordingly, some agents that have potential for in vivo use (e.g., use in treating, diagnosing or preventing disease) have only limited efficacy because they are rapidly degraded and inactivated by proteases.

Targeting VEGF with currently available therapeutics is not effective in all patients, or for all cancers.

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