Heterodimerized polypeptide

Patent metadata

Field	Value
Publication number	US-12371511-B2
Application number	US-201815860163-A
Country	US
Kind code	B2
Filing date	Jan 2, 2018
Priority date	Jun 30, 2011
Publication date	Jul 29, 2025
Grant date	Jul 29, 2025

Abstract

The present inventors produced a heterodimerized polypeptide having an Fc region formed from two polypeptides with different amino acid sequences (a first polypeptide and a second polypeptide), and succeeded in producing a heterodimerized polypeptide containing an Fc region with improved Fc region function compared to that of a homodimer in which the Fc region is composed of only the first polypeptide or only the second polypeptide by conventional technology.

First claim

The invention claimed is:

1. A molecule comprising a heterodimeric Fc region that is a heterodimer of a first Fc polypeptide and a second Fc polypeptide that differs in amino acid sequence from the first Fc polypeptide,
wherein the first Fc polypeptide comprises a hinge region or a part thereof, a first human IgG1 CH2 domain and a CH3 domain, wherein the first human IgG1 CH2 domain comprises a set of substitutions selected from sets (i) to (vi), and wherein all positions are by EU numbering:

(i) substitution of Leu at position 234 with Tyr, Ala, Glu, Gly, His, or Ser,
substitution of Leu at position 235 with Tyr, Ala, Glu, Phe, Ile, Leu, Met, Asn, Pro, Gln, Thr, Val, or Trp,
substitution of Gly at position 236 with Trp or Tyr,
substitution of His at position 268 with Asp, Glu, or Ala, and
substitution of Ser at position 298 with Ala;

(ii) substitution of Leu at position 234 with Tyr, Ala, Glu, Gly, His, or Ser,
substitution of Leu at position 235 with Tyr, Ala, Glu, Phe, Ile, Leu, Met, Asn, Pro, Gln, Thr, Val, or Trp,
substitution of Gly at position 236 with Trp or Tyr,
substitution of His at position 268 with Asp, Glu, or Ala,
substitution of Asp at position 270 with Glu, and
substitution of Ser at position 298 with Ala;

(iii) substitution of Leu at position 234 with Tyr, Ala, Glu, Gly, His, or Ser,
substitution of Leu at position 235 with Tyr, Ala, Glu, Phe, Ile, Leu, Met, Asn, Pro, Gln, Thr, Val, or Trp,
substitution of Gly at position 236 with Trp or Tyr,
substitution of Ser at position 239 with Met,
substitution of His at position 268 with Asp, Glu, or Ala,
substitution of Asp at position 270 with Glu, and
substitution of Ser at position 298 with Ala;

(iv) substitution of Leu at position 234 with Tyr, Ala, Glu, Gly, His, or Ser,
substitution of Leu at position 235 with Tyr, Ala, Glu, Phe, Ile, Leu, Met, Asn, Pro, Gln, Thr, Val, or Trp,
substitution of Gly at position 236 with Trp or Tyr,
substitution of His at position 268 with Asp, Glu, or Ala,
substitution of Ser at position 298 with Ala, and
substitution of Ala at position 327 with Asp;

(v) substitution of Leu at position 234 with Tyr, Ala, Glu, Gly, His, or Ser,
substitution of Leu at position 235 with Tyr, Ala, Glu, Phe, Ile, Leu, Met, Asn, Pro, Gln, Thr, Val, or Trp,
substitution of Gly at position 236 with Trp or Tyr,
substitution of Ser at position 239 with Met,
substitution of His at position 268 with Asp, Glu, or Ala,
substitution of Ser at position 298 with Ala, and
substitution of Ala at position 327 with Asp;

(vi) substitution of Leu at position 234 with Tyr, Ala, Glu, Gly, His, or Ser,
substitution of Leu at position 235 with Tyr, Ala, Glu, Phe, Ile, Leu, Met, Asn, Pro, Gln, Thr, Val, or Trp,
substitution of Gly at position 236 with Trp or Tyr,
substitution of Ser at position 239 with Met,
substitution of His at position 268 with Asp, Glu, or Ala,
substitution of Ser at position 298 with Ala,
substitution of Ala at position 327 with Asp,
substitution of Leu at position 328 with Trp, and
substitution of Lys at position 334 with Ile, Glu, or Asp;

and wherein the second Fc polypeptide comprises a hinge region or a part thereof, a second human IgG1 CH2 domain and a CH3 domain, wherein the second human IgG1 CH2 domain comprises a set of substitutions selected from sets (vii) and (viii), and wherein all positions are by EU numbering:
(vii) substitution of Lys at position 326 with Thr, Asp, or Ile,
substitution of Ala at position 330 with Lys, Pro, Met, or Phe, and
substitution of Lys at position 334 with Ile, Glu, or Asp;

(viii) substitution of Asp at position 270 with Glu,
substitution of Lys at position 326 with Thr, Asp, or Ile,
substitution of Ala at position 330 with Lys, Pro, Met, or Phe, and
substitution of Lys at position 334 with Ile, Glu, or Asp.

2. The molecule of claim 1 , wherein the molecule is an antibody.

3. The molecule of claim 2 , wherein the antibody is a bispecific antibody.

4. The molecule of claim 1 , wherein the molecule is a peptide Fc fusion protein or a scaffold Fc fusion protein.

5. A pharmaceutical composition comprising the molecule of claim 1 and a medically acceptable carrier.

6. A method of producing the molecule of claim 1 , the method comprising:
identifying a parent molecule comprising an Fc region that is a homodimer of a starting polypeptide;
producing a first mutated version of the starting polypeptide, the first mutated version being the first polypeptide;
producing a second mutated version of the starting polypeptide, the second mutated version being the second polypeptide; and
generating the molecule comprising the heterodimeric Fc region that is a heterodimer of the first polypeptide and the second polypeptide.

7. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (i).

8. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (ii).

9. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (iii).

10. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (iv).

11. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (v).

12. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (vi).

13. The molecule of claim 1 , wherein the second human IgG1 CH2 domain comprises the substitutions of set (vii).

14. The molecule of claim 1 , wherein the second human IgG1 CH2 domain comprises the substitutions of set (viii).

15. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (i), and the second human IgG1 CH2 domain comprises the substitutions of set (vii).

16. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (i), and the second human IgG1 CH2 domain comprises the substitutions of set (viii).

17. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (ii), and the second human IgG1 CH2 domain comprises the substitutions of set (vii).

18. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (ii), and the second human IgG1 CH2 domain comprises the substitutions of set (viii).

19. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (iii), and the second human IgG1 CH2 domain comprises the substitutions of set (vii).

20. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (iii), and the second human IgG1 CH2 domain comprises the substitutions of set (viii).

21. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (iv), and the second human IgG1 CH2 domain comprises the substitutions of set (vii).

22. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (iv), and the second human IgG1 CH2 domain comprises the substitutions of set (viii).

23. The molecule of claim 1 , wherein the first human IgG1 CH2 domain comprises the substitutions of set (v), and the second human IgG1 CH2 domain comprises the substitutions of set (vii).

24. The molecule of claim

Assignees

• Chugai Pharmaceutical Co Ltd

Inventors

• Mimoto Futa
• Kuramochi Taichi
• Igawa Tomoyuki
• Kawazoe Meiri
• Katada Hitoshi
• Shiraiwa Hirotake
• Kadono Shojiro

Classifications

• G01N33/566

  using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds} · CPC title

• C07K1/1075 Primary

  by covalent attachment of amino acids or peptide residues · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K2317/732

  Antibody-dependent cellular cytotoxicity [ADCC] · CPC title