US 7,597,889 B1 · Binding molecules derived from immunoglobulins which do not trigger complement mediated lysis

Abstract

Disclosed are binding molecules which are recombinant polypeptides containing: (i) a binding domain capable of binding a target molecule, and (ii) an effector domain having an amino acid sequence substantially homologous to all or part of a constant domain of a human immunoglobulin heavy chain; characterized in that the binding molecule is capable of binding the target molecule without triggering significant complement dependent lysis, or cell mediated destruction of the target, and more preferably wherein the effector domain is capable of specifically binding FcRn and/or FcγRIIb. These are generally based on chimeric domains which are derived from two or more human immunoglobulin heavy chain CH2 domains domains. In preferred embodiments the regions 233-236, and 327-331, are modified, as are further residues to render the molecule null allotypic. Also disclosed are nucleic acids, host cells, production processes and materials, and uses. Pharmaceutical preparations are also disclosed.

Claims52

Dependent claims

A binding molecule which is a recombinant polypeptide comprising:

(i) a binding domain capable of binding a target molecule, which binding domain is the binding site of an antibody, and
(ii) an effector domain having an amino acid sequence homologous to a constant domain of a human immunoglobulin heavy chain;
wherein the binding molecule is capable of binding the target molecule without triggering significant complement dependent lysis, or cell mediated destruction of the target, and the effector domain is capable of specifically binding FcγRIIb and optionally FcRn,
and wherein the effector domain comprises a chimeric CH2 domain which is derived from two or more human immunoglobulin heavy chain CH2 domains, which human immunoglobulins are selected from IgG1, IgG2 and IgG4,
and wherein the effector domain has a reduced affinity for FcγRI, FcγRIIa and FcγRIII and a reduced ability to mediate complement lysis by comparison with said constant domain of a human immunoglobulin heavy chain
and wherein the chimeric CH2 domain is a human immunoglobulin heavy chain CH2 domain which has the following blocks of amino acids at the stated positions:
233P, 234V, 235A, 236G, 327G, 330S and 331S numbered with respect to the EU numbering system of Kabat, and is at least 98% identical to G1 Δac (SEQ ID NO:3) or G4 Δc (SEQ ID NO:12) as shown in FIG. 17.

4 more elements

A binding molecule which is a recombinant polypeptide comprising:

(i) a binding domain capable of binding a target molecule, which binding domain is the binding site of an antibody, and
(ii) an effector domain having an amino acid sequence homologous to a constant domain of a human immunoglobulin heavy chain;
wherein the binding molecule is capable of binding the target molecule without triggering significant complement dependent lysis, or cell mediated destruction of the target, and the effector domain is capable of specifically binding FcγRIIb and optionally FcRn,
and wherein the effector domain comprises a chimeric CH2 domain which is derived from two or more human immunoglobulin heavy chain CH2 domains, which human immunoglobulins are selected from IgG1, IgG2 and IgG4,
and wherein the effector domain has a reduced affinity for FcγRI, FcγRIIa and FcγRIII and a reduced ability to mediate complement lysis by comparison with said constant domain of a human immunoglobulin heavy chain
and wherein the chimeric CH2 domain is a human immunoglobulin heavy chain CH2 domain which has the following blocks of amino acids at the stated positions:
233P, 234V, 235A and no residue at 236, 327G, 330S and 331S, numbered with respect to the EU system of Kabat, and is at least 98% identical to G1 Δab (SEQ ID NO:1) or G2 Δa (SEQ ID NO:2) as shown in FIG. 17.

4 more elements

A process for producing a binding molecule which is a recombinant polypeptide comprising:

(i) a binding domain capable of binding a target molecule, which binding domain is the binding site of an antibody, and
(ii) an effector domain having an amino acid sequence homologous to a constant domain of a human immunoglobulin heavy chain;
wherein the binding molecule is capable of binding the target molecule without triggering significant complement dependent lysis, or cell mediated destruction of the target, and the effector domain is capable of specifically binding FcγRIIb and optionally FcRn,
and wherein the effector domain comprises a chimeric CH2 domain which is derived from two or more human immunoglobulin heavy chain CH2 domains, which human immunoglobulins are selected from IgG1, IgG2 and IgG4,
and wherein the effector domain has a reduced affinity for FcγRI, FcγRIIa and FcγRIII and a reduced ability to mediate complement lysis by comparison with said constant domain of a human immunoglobulin heavy chain;
the process comprising the steps of modifying a nucleotide sequence encoding a first human immunoglobulin heavy chain CH2 domain such that 2, 3 or 4 amino acids in at least 1 region of the CH2 domain correspond to the amino acids from a second human immunoglobulin heavy chain CH2 domain,
wherein said modification introduces the following blocks of amino acids at the stated positions:
233P, 234V, 235A and no residue at 236 and 327G, 330S and 331S numbered with respect to the ELU numbering system of Kabat
and wherein said chimeric CH2 domain is at least 98% identical to G1 Δab (SEQ ID NO:1) or G2 Δa (SEQ ID NO:2) as shown in FIG. 17,
introducing into a host cell a vector comprising said modified nucleotide sequence,
culturing said host cell under conditions such that said binding molecule is produced, and
isolating said binding molecule from said cell culture.

9 more elements

Dependencies are inferred from the claim text.

Binding molecules derived from immunoglobulins which do not trigger complement mediated lysis

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Binding molecules derived from immunoglobulins which do not trigger complement mediated lysis

Life of this patent

Abstract

Claims52