Bispecific antibodies and uses thereof

Abstract

Methods, compositions and uses are provided for bispecific antibodies comprising one or more unnatural amino acids. The bispecific antibodies may bind to two or more different receptors, co-receptors, antigens, or cell markers on one or more cells. The bispecific antibodies may be used to treat a disease or condition (e.g., cancer, autoimmune disease, pathogenic infection, inflammatory disease). The bispecific antibodies may be used to modulate (e.g., stimulate or suppress) an immune response.

Description

CROSS-REFERENCE[0001]This application claims the benefit of U.S. provisional application Ser. No. 61 / 783,426, filed Mar. 14, 2013 and U.S. provisional application Ser. No. 61 / 839,330, filed Jun. 25, 2013; which are incorporated by reference in their entirety.STATEMENT AS TO FEDERALLY SPONSORED RESEARCH[0002]This invention was made with the support of the United States government under Contract number GM062159 by National Institutes of Health (NIH). The government has certain rights in the invention.FIELD OF THE INVENTION[0003]Described herein are immunoglobulin constructs comprising one or more unnatural amino acids, methods of making such constructs, pharmaceutical compositions and medicaments comprising such constructs, and methods of using such constructs and compositions to prevent, inhibit, and / or treat a disease or condition in a subject.BACKGROUND OF THE INVENTION[0004]Antibodies are natural proteins that the vertebrate immune system forms in response to foreign substances (a...

AI Technical Summary

Benefits of technology

[0013]Disclosed herein are methods of producing a bispecific antibodies of Formula II: X-L1′-L2′-Y or Formula IIA: Y-L2′-L1′-X, comprising: coupling L1″ to X to produce a first intermediate of Formula III: X-L1′ or Formula IIIA: L1′-X, wherein X comprises an anti-CD3 antibody or anti-CD3 antibody fragment; and L1″ comprises a first linker before being coupled to X, coupling L2″ to Y to produce a second intermediate of Formula IV: Y-L2′ or Formula IVA: L2′-Y; wherein Y comprises at least a portion of a second antibody or antibody fragment and L2″ comprises a second linker before being coupled to Y; and linking the first intermediate to the second intermediate, thereby producing the antibody of Formula II or HA. Coupling L1″ to X may comprise site-specific coupling of L1″ to X. Coupling L2″ to Y may comprise site-specific coupling of L2″ to Y. The method may further comprise incorporating one or more unnatural amino acids into X. The method may further comprise incorporating one or more unnatural amino acids into Y. Incorporating one or more unnatural amino acids into X and/or Y may comprise modifying one or more amino acid residues in X and/or Y to produce one or more amber codons in X and/or Y. The one or more unnatural amino acids may be incorporated into X and/or Y in response to an amber codon. The one or more unnatural amino acids may be site-specifically incorporated into X and/or Y. Incorporating one or more unnatural amino acids may comprise the use of one or more tRNA synthetases. The tRNA synthetase may be an aminoacyl tRNA synthetase. The tRNA synthetase may be a mutant tRNA synthesis. Incorporating one or more unnatural amino acids may comprise a tRNA/tRNA synthetase pair. The tRNA/tRNA synthetase pair may comprise a tRNA/aminoacyl-tRNA synthetase pair. The tRNA/tRNA synthetase pair may comprise a tRNATyr/tyrosyl-tRNA synthetase pair.

[0014]Coupling L1″ to X may occur at the one or more unnatural amino acids in X. Coupling L2″ to Y may occur at the one or more unnatural amino acids in Y. Coupling L1″ to X and coupling L2″ to Y may occur sequentially. Coupling L1″ to X and coupling L2 to Y may occur simultaneously. Coupling L1″ to X may occur prior to coupling L2″ to Y. Coupling L2″ to Y may occur prior to coupling L1″ to X. Coupling L1″ to X and/or coupling L2″ to Y may comprise forming one or more oxime bonds between L1″ and X and/or between L2″ and Y. Coupling L1″ to X and/or coupling L2″ to Y may comprise forming one or more stable bonds between L1″ and X and/or between L2″ and Y. Coupling L1″ to X and/or coupling L2″ to Y may comprise forming one or more covalent bonds between L1″ and X and/or between L2″ and Y. Coupling L1″ to X and/or coupling L2″ to Y may comprise forming one or more non-covalent bonds between L1″ and X and/or between L2″ and Y. Coupling L1″ to X and/or coupling L2″ to Y may comprise forming one or more ionic bonds between L1″ and X and/or between L2″ and Y.

[0015]Linking the first intermediate to the second intermediate may comprise a Huisgen-cycloaddition. Linking the first intermediate to the second intermediate may comprise a Diels-Halder reaction Linking the first intermediate to the second intermediate may comprise a hetero Diels-Alder reaction Linking the first intermediate to the second intermediate may comprise an enzyme-mediated reaction Linking the first intermediate to the second intermediate may comprise a copper-free [3+2] Huisgen-cycloaddition reaction Linking the first intermediate to the second intermediate may produce an oxime Linking the first intermediate to the second intermediate may produce a tetrazole Linking the first intermediate to the second intermediate may produce a Diels Alder adduct Linking the first intermediate to the second intermediate may produce a hetero Diels Alder adduct Linking the first intermediate to the second intermediate may produce an aromatic substitution reaction product Linking the first intermediate to the second intermediate may produce a nucleophilic substitution reaction product Linking the first intermediate to the second intermediate may produce an ester Linking the first intermediate to the second intermediate may produce an amide Linking the first intermediate to the second intermediate may produce a carbamate Linking the first intermediate to the second

Problems solved by technology

These current approaches may result in heterogeneous products and/or may require the development of antigen-spec

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