Modified Fc molecules

摘要

Disclosed is a process for preparing a pharmacologically active compound, in which at least one internal conjugation site of an Fc domain sequence is selected that is amenable to conjugation of an additional functional moiety by a defined conjugation chemistry through the side chain of an amino acid residue at the conjugation site. An appropriate amino acid residue for conjugation may be present in a native Fc domain at the conjugation site or may be added by insertion (i.e., between amino acids in the native Fc domain) or by replacement (i.e., removing amino acids and substituting different amino acids). In the latter case, the number of amino acids added need not correspond to the number of amino acids removed from the previously existing Fc domain. This technology may be used to produce useful compositions of matter and pharmaceutical compositions containing them. A DNA encoding the inventive composition of matter, an expression vector containing the DNA, and a host cell containing the expression vector are also disclosed.

主权项

1. A composition of matter of the formula wherein: F1 is a monomer of the monomeric or multimeric Fc domain; X1 is covalently bound to the N-terminus of F1 through the α-amino site of F1; X2 is covalently bound to the C-terminus of F1 through the α-carboxy site of F1; X3 is covalently bound to the one or more specifically selected internal conjugation site(s) in F1 through the side chain of an amino acid residue selected from the group consisting of D46, S48, H49, E50, E53, K55, D61, G62, Q76, K107, K121, G122, Q123, R136, D137, T140, K141, N142, N170, K173, S181, G183, D194, K195, R197, and Q199, relative to reference sequence SEQ ID NO: 599, or, if g>1, any combination of these members; X1, X2, and X3 are each independently selected from -(L1)c-P0, -(L1)c-P1, -(L1)c-P1-(L2)d-P2, -(L1)c-P1-(L2)d-P2-(L3)e-P3, and -(L1)c-P1-(L2)d-P2-(L3)e-P3-(L4)f-P4; P0, P1, P2, P3, and P4 are each independently selected from the group consisting of:
i) a pharmaceutically acceptable polymer or dextran;ii) a pharmacologically active polypeptide, peptide, peptidomimetic, or non-peptide organic moiety;iii) a radioisotope, an enzyme, a biotinyl moiety, a fluorophore, or a chromophore; and iv) an immobilized substrate, provided that in a chain comprising more than one additional functional moieties, the immobilized substrate is the moiety most distal from F1, and there can be no more than one immobilized substrate in the chain; L1, L2, L3, and L4 are each independently linkers; a, b, c, d, e, and f are each independently 0 or 1; and g is 1, 2, 3, or 4.