Engineered immunoglobulin heavy chain-light chain pairs and uses thereof

Patent metadata

Field	Value
Publication number	US-10077298-B2
Application number	US-201314648222-A
Country	US
Kind code	B2
Filing date	Nov 28, 2013
Priority date	Nov 28, 2012
Publication date	Sep 18, 2018
Grant date	Sep 18, 2018

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 C H1 and/or C L domains, one or more amino acid modifications in the V H and/or V L 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.

First claim

We claim:

1. An isolated antigen binding polypeptide construct comprising at least a first heterodimer and a second heterodimer,
the first heterodimer comprising a first immunoglobulin heavy chain polypeptide sequence (H1) and a first immunoglobulin light chain polypeptide sequence (L1), and binding to a first epitope; and the second heterodimer comprising a second immunoglobulin heavy chain polypeptide sequence (H2) and a second immunoglobulin light chain polypeptide sequence (L2), and binding to a second epitope,
wherein at least one of the H1 or L1 sequences of the first heterodimer is distinct from the corresponding H2 or L2 sequence of the second heterodimer, H1 and H2 each comprise a heavy chain variable domain (V H domain) and a heavy chain constant domain (C H1 domain), and L1 and L2 each comprise a light chain variable domain (V L domain) and a light chain constant domain (C L domain);
wherein:
i) H1 comprises amino acid substitutions at residues 143 and 145, L1 comprises amino acid substitution at residue 124, H2 comprises amino acid substitutions at residues 146 and 179, and L2 comprises amino acid substitution at residue 124, wherein the amino acid substitution at residue 143 is 143E or 143D, the amino acid substitution at residue 145 is 145T, or conservative substitution thereof, the amino acid substitution at residue 124 of L1 is 124R or 124K, the amino acid substitution at residue 146 is 146G, or conservative substitution thereof, the amino acid substitution at residue 179 is 179R or 179K, and the amino acid substitution at residue 124 of L2 is 124E or 124D; or
ii) H1 comprises amino acid substitutions at residues 145 and 179, L1 comprises amino acid substitution at residue 131, H2 comprises amino acid substitution at residue 186, and L2 comprises amino acid substitution at residues 160 and 180, wherein the amino acid substitution at residue 145 is 145T or 145L or conservative substitutions thereof, the amino acid substitution at residue 179 is 179E or 179D, the amino acid substitution at residue 131 is 131K or 131R, the amino acid substitution at residue 186 is 186K or 186R, the amino acid substitution at residue 160 is 160E or 160D, and the amino acid substitution at residue 180 is 180E or 180D;
and wherein the numbering of amino acid residues is according to Kabat.

2. The construct of claim 1 , 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-L2 in the absence of the amino acid substitutions.

3. The construct of claim 1 , wherein H1 comprises amino acid substitutions at residues 143 and 145, L1 comprises amino acid substitution at residue 124, H2 comprises amino acid substitutions at residues 146 and 179, and L2 comprises amino acid substitution at residue 124, wherein the amino acid substitution at residue 143 is 143E or 143D, the amino acid substitution at residue 145 is 145T, or conservative substitution thereof, the amino acid substitution at residue 124 of L1 is 124R or 124K, the amino acid substitution at residue 146 is 146G, or conservative substitution thereof, the amino acid substitution at residue 179 is 179R or 179K, and the amino acid substitution at residue 124 of L2 is 124E or 124D, and
L2 further comprises amino acid substitutions at residues 160 and 180, and the amino acid substitution at residue 160 is 160E or 160D, and the amino acid substitution at residue 180 is 180E or 180D.

4. The construct of claim 1 , wherein H1 comprises amino acid substitutions at residues 143 and 145, L1 comprises amino acid substitution at residue 124, H2 comprises amino acid substitutions at residues 146 and 179, and L2 comprises amino acid substitution at residue 124, wherein the amino acid substitution at residue 143 is 143E or 143D, the amino acid substitution at residue 145 is 145T, or conservative substitution thereof, the amino acid substitution at residue 124 of L1 is 124R or 124K, the amino acid substitution at residue 146 is 146G, or conservative substitution thereof, the amino acid substitution at residue 179 is 179R or 179K, and the amino acid substitution at residue 124 of L2 is 124E or 124D, and
L2 further comprises amino acid substitutions at residues 160 and 178, and the amino acid substitution at residue 160 is 160E or 160D, and the amino acid substitution at residue 178 is 178E or 178D.

5. The construct of claim 1 , wherein H1 comprises amino acid substitutions at residues 143 and 145, L1 comprises amino acid substitution at residue 124, H2 comprises amino acid substitutions at residues 146 and 179, and L2 comprises amino acid substitution at residue 124, wherein the amino acid substitution at residue 143 is 143E or 143D, the amino acid substitution at residue 145 is 145T, or conservative substitution thereof, the amino acid substitution at residue 124 of L1 is 124R or 124K, the amino acid substitution at residue 146 is 146G, or conservative substitution thereof, the amino acid substitution at residue 179 is 179R or 179K, and the amino acid substitution at residue 124 of L2 is 124E or 124D, and
L1 further comprises amino acid substitutions at residues 160 and 178, and the amino acid substitution at residue 160 is 160K or 160R, and the amino acid substitution at residue 178 is 178R or 178K.

6. The construct of claim 1 , wherein:
a. H1 comprises amino acid substitutions 143E and 145T; L1 comprises amino acid substitution 124R; H2 comprises amino acid substitutions 146G and 179K; and L2 comprises amino acid substitutions 124E and 160E and 180E;
b. H1 comprises amino acid substitutions 143E and 145T; L1 comprises amino acid substitutions 124R and 160K and 178R; H2 comprises amino acid substitutions 146G and 179K; and L2 comprises amino acid substitutions 124E and 160E and 180E, or
c. H1 comprises amino acid substitutions 143E and 145T; L1 comprises amino acid substitutions 124R and 160K and 178R; H2 comprises amino acid substitutions 146G and 179R; and L2 comprises amino acid substitutions 124E and 160E and 178D.

7. The construct of claim 1 , wherein H1 comprises amino acid substitutions at residues 145 and 179, L1 comprises amino acid substitution at residue 131, H2 comprises amino acid substitution at residue 186, and L2 comprises amino acid substitution at residues 160 and 180, wherein the amino acid substitution at residue 145 is 145T or 145L or conservative substitutions thereof, the amino acid substitution at residue 179 is 179E or 179D, the amino acid substitution at residue 131 is 131K or 131R, the amino acid substitution at residue 186 is 186K or 186R, the amino acid substitution at residue 160 is 160E or 160D, and the amino acid substitution at residue 180 is 180E or 180D, and
L2 further comprises amino acid substitution at residue 124, wherein the amino acid substitution at residue 124 is 124E or 124D.

8. The construct of claim 1 , wherein:
a. H1 comprises 145T and 179E, L1 comprises 131K, H2 comprises 186R and L2 comprises 160E and 180E;
b. H1 comprises 145T and 179E, L1 comprises 131K, H2 comprises 186R and L2 comprises 124E, 160E and 180E or
c. H1 comprises 145L and 179E, L1 comprises 131K, H2 comprises 186R and L2 comprises 124E, 160E and 180E.

9. The construct of claim 1 , wherein H1, H2, L1 and L2 are co-expressed in a cell or a mammalian cell, or H1, H2, L1 and L2 are co-expressed in a cell-free expression system, or H1, H2, L1 and L2 are co-produced, or H1, H2, L1 and L2 are co-produced via a redox production system.

10. The construct of claim 1 , wherein the first immunoglobulin light chain polypeptide sequence and/or the second immunoglobulin light chain polypeptide sequence is a kappa light cha

Assignees

• Zymeworks Inc

Inventors

• Corper Adam Louis
• Urosev Dunja
• Tom-Yew Stacey A L
• Bleile Dustin Weyland Blue
• Spreter Von Kreudenstein Thomas
• Dixit Surjit
• Lario Paula Irene
• Sanches Mario

Classifications

• A61P9/14

  Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• G16B40/00

  ICT specially adapted for biostatistics; ICT specially adapted for bioinformatics-related machine learning or data mining, e.g. knowledge discovery or pattern finding · CPC title