US 11,306,156 B2 · Modified antigen binding polypeptide constructs and uses thereof

Abstract

The present invention provides heterodimer pairs that can comprise a first heterodimer and a second heterodimer wherein each heterodimer comprises an immunoglobulin heavy chain or fragment thereof and an immunoglobulin light chain or fragment thereof. At least one of the heterodimers can comprise one or more amino acid modifications in the CH1 and/or CL domains, one or more amino acid modifications in the VH and/or VL domains, or a combination thereof. The modified amino acid(s) can be part of the interface between the light chain and heavy chain and are typically modified to create preferential pairing between each heavy chain and a desired light chain such that when the two heavy chains and two light chains of the heterodimer pair are co-expressed in a cell, the heavy chain of the first heterodimer preferentially pairs with one of the light chains rather than the other. Likewise, the heavy chain of the second heterodimer typically preferentially pairs with the second light chain rather than first.

An antigen binding-polypeptide construct comprising at least a first heterodimer and a second heterodimer,

the first heterodimer comprising a first human or humanized immunoglobulin G (IgG) heavy chain polypeptide sequence (H1), and a first human or humanized immunoglobulin kappa light chain polypeptide sequence (L1), and binding to a first epitope;
and the second heterodimer comprising a second human or humanized immunoglobulin G (IgG) heavy chain polypeptide sequence (H2), and a second human or humanized immunoglobulin kappa light chain polypeptide sequence (L2), and binding to a second epitope, wherein the H1 and L1 polypeptide sequences of the first heterodimer are different from the corresponding H2 and L2 polypeptide sequences of the second heterodimer,
wherein H1 and H2 each comprise a heavy chain variable domain (VH domain) and a heavy chain constant domain (CH1 domain);
wherein L1 and L2 each comprise a light chain variable domain (VL domain) and a light chain constant domain (CL domain);
wherein H1, H2, L1, and L2 comprise the following amino acid substitutions that promote preferential pairing of H1 with L1 as compared to L2 and of H2 with L2 as compared to L1, at positions identified according to the Kabat numbering system:
a) H1 comprises amino acid substitutions 143E, 145T, 179E, and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 176L, and 180E,
b) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 178E, and 180E,
c) H1 comprises amino acid substitutions 143E, 145T, 179E, and 188L, L1 comprises amino acid substitution 124R and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 131T, 178Y, and 180E,
d) H1 comprises amino acid substitutions 143E, 145T, 179E, and 188L, L1 comprises amino acid substitution 124R and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 131T, 178F, and 180E,
e) H1 comprises amino acid substitutions 143E, 145T, and 188L, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 131T, 178Y, and 180E,
f) H1 comprises amino acid substitutions 143E, 145T, and 188L, L1 comprises amino acid substitution 124R, 160K and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 131T, 178F, and 180E,
g) H1 comprises amino acid substitutions 143E, 145T, and 188L, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179K, and L2 comprises amino acid substitutions 124E, 176L, and 180E,
h) H1 comprises amino acid substitutions 143E, 145T, 179E, and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E, 176L, and 180E,
i) H1 comprises amino acid substitutions 143E, 145T, 179E, and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E, 131T, 178Y, and 180E,
j) H1 comprises amino acid substitutions 143E, 145T, 179E, and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E, 131T, 178F, and 180E,
k) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E, 178E, and 180E,
l) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E and 178E,
m) H1 comprises amino acid substitutions 143E, 145T, and 188L, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E, 176L, and 180E,
n) H1 comprises amino acid substitutions 143E, 145T, and 188L, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 179R, and L2 comprises amino acid substitutions 124E, 131T, 178Y, and 180E,
o) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 160E and 178E,
p) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E and 178E,
q) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 178E and 180E,
r) H1 comprises amino acid substitutions 143E, 145T, 179E and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 131T, 178F and 180E,
s) H1 comprises amino acid substitutions 143E, 145T, 179E and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 131T, 178Y and 180E,
t) H1 comprises amino acid substitutions 143E, 145T, 179E and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 176L and 180E,
u) H1 comprises amino acid substitutions 143E, 145T and 188L, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 131T, 178Y and 180E,
v) H1 comprises amino acid substitutions 143E, 145T and 188L, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 186K, and L2 comprises amino acid substitutions 124E, 176L and 180E,
w) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, 160K and 178R, H2 comprises amino acid substitution 186R, and L2 comprises amino acid substitutions 124E, 178E and 180E,
x) H1 comprises amino acid substitutions 143E, 145T, 179E and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 186R, and L2 comprises amino acid substitutions 124E, 131T, 178Y and 180E, or
y) H1 comprises amino acid substitutions 143E, 145T, 179E and 188L, L1 comprises amino acid substitutions 124R and 178R, H2 comprises amino acid substitution 186R, and L2 comprises amino acid substitutions 124E, 176L and 180E,
wherein when both L1 and L2 are co-expressed with at least one of H1 and H2, the pairing of H1-L1 to H1-L2 and the pairing of H2-L2 to H2-L1 is greater than the pairing of H1-L1 to H1-L2 and the pairing of H2-L2 to H2-L1 in the absence of the amino acid substitutions.

28 more elements

Modified antigen binding polypeptide constructs and uses thereof

Life of this patent

Modified antigen binding polypeptide constructs and uses thereof

Life of this patent

Abstract

Claims37