HETERODIMERIZED POLYPEPTIDE

• US 20150344570A1
• Filed: 12/26/2013
• Published: 12/03/2015
• Est. Priority Date: 12/27/2012
• Status: Active Grant

• Current Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Igawa, Tomoyuki, Mimoto, Futa, Katada, Hitoshi, Shiraiwa, Hirotake
• Granted Patent: US 10,766,960 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/303 Liver or Pancreas
• CPC Class Codes: C07K 16/46 Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/72 Increased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies

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 function compared to that of a homodimer in which the Fc region is composed of only the first polypeptide or the second polypeptide by conventional technology.

Claims

1. A polypeptide characterized in that the polypeptide is composed of a heterodimer comprising a first polypeptide and a second polypeptide, wherein either one of the first polypeptide and the second polypeptide comprises an Fc region introduced with the mutations of (i) or (ii), which is a polypeptide whose function of the Fc region is altered compared to a polypeptide comprising an Fc region without mutations:

(i) according to EU numbering, the amino acid at position 234 is L, S, F, E, V, D, Q, I, M, T, A, G, or H;

the amino acid at position 235 is Y or Q;

the amino acid at position 236 is W;

the amino acid at position 239 is M or I;

the amino acid at position 268 is D; and

the amino acid at position 298 is A;

(ii) according to EU numbering, the amino acid at position 270 is E;

the amino acid at position 326 is D;

the amino acid at position 330 is A, K, M, F, I, Y, or H; and

the amino acid at position 334 is E.

2. The polypeptide of claim 1, wherein either the first polypeptide or the second polypeptide comprises an Fc region introduced with the mutations of (i) or (ii), and the mutations of (iii) are introduced into the other polypeptide:

(i) according to EU numbering, the amino acid at position 234 is L, S, F, E, V, D, Q, I, M, T, A, G or H;

the amino acid at position 235 is Y or Q;

the amino acid at position 236 is W;

the amino acid at position 239 is M or I;

the amino acid at position 268 is D;

the amino acid at position 298 is A; and

the amino acid at position 327 is D;

(ii) according to EU numbering, the amino acid at position 234 is L, S, F, E, V, D, Q, I, M, T, A, G or H;

the amino acid at position 235 is Y or Q;

the amino acid at position 236 is W;

the amino acid at position 239 is M or I;

the amino acid at position 268 is D;

the amino acid at position 270 is E; and

the amino acid at position 298 is A;

(iii) according to EU numbering, the amino acid at position 270 is E;

the amino acid at position 326 is D;

the amino acid at position 330 is A, K, M, F, I, Y, or H; and

the amino acid at position 334 is E.

3. The polypeptide of claim 2, wherein in claim 2 (i), the amino acid at position 234 according to EU numbering is E, D, T, or L.

4. The polypeptide of claim 2, wherein in claim 2 (ii), the amino acid at position 234 according to EU numbering is L, F, E, or D.

5. The polypeptide of claim 2, wherein in claim 2 (i), the amino acid at position 234 according to EU numbering is V, I, T, M, or L.

6. The polypeptide of claim 2, wherein in claim 2 (i), the amino acid at position 234 according to EU numbering is V, E, D, T, I, L, or F, and the amino acid at position 239 is M or I;

and in (iii), the amino acid at position 330 according to EU numbering is A or K.

7. The polypeptide of claim 2, wherein in claim 2 (ii), the amino acid at position 234 according to EU numbering is F, E, D, S, or L, and the amino acid at position 239 is M or I;

and in (iii), the amino acid at position 330 according to EU numbering is A, F, or K.

8. The polypeptide of claim 1, wherein the alteration of function of the Fc region is at least one or more alterations selected from the group consisting of enhancement of binding activity, weakening of binding activity, and improvement of selectivity of binding activity of the polypeptide to an Fcγ

receptor(s).

9. The polypeptide of claim 8, wherein the Fcγ

receptor(s) is at least one or more receptors selected from the group consisting of Fcγ

RIa, Fcγ

RIIa R, Fcγ

RIIa H, Fcγ

RIIb, Fcγ

RIIIaF, and Fcγ

RIIIaV.

10. The polypeptide of claim 8, wherein the alteration of function of the Fc region is improvement of selectivity of binding activity to an Fcγ

receptor(s).

11. The polypeptide of claim 10, wherein the improvement of selectivity of binding activity to an Fcγ

receptor(s) refers to selectivity between an activating Fcγ

receptor(s) and an inhibitory Fcγ

receptor, and wherein among the aforementioned Fcγ

receptors, the activating Fcγ

receptor(s) is at least one or more receptors selected from the group consisting of Fcγ

RIa, Fcγ

RIIa R, Fcγ

RIIa H, Fcγ

RIIIaF, and Fcγ

RIIIaV, and the inhibitory Fcγ

receptor is Fcγ

RIIb.

13. The polypeptide of claim 11, wherein the binding activity to the activating Fcγ

receptor(s) is selectively enhanced compared to the binding activity to the inhibitory Fcγ

receptor.

14. The polypeptide of claim 1, wherein an amino acid alteration for conferring difference to the isoelectric points of the first polypeptide and the second polypeptide is further introduced.

15. The polypeptide of claim 14, wherein the amino acid alteration for conferring difference to isoelectric points is characterized by the introduction of at least one amino acid mutation at an amino acid site selected from the group consisting of Gly at position 137, Gly at position 138, Thr at position 139, Lys at position 147, Ser at position 192, Leu at position 193, Gln at position 196, Tyr at position 198, Ile at position 199, Asn at position 203, Lys at position 214, Ala at position 231, Glu at position 233, Leu at position 242, Val at position 263, Glu at position 272, Lys at position 274, Tyr at position 278, Lys at position 288, Lys at position 290, Gly at position 316, Lys at position 317, Lys at position 320, Lys at position 324, Thr at position 335, Ser at position 337, Lys at position 338, Lys at position 340, Gly at position 341, Leu at position 358, Lys at position 360, Gln at position 362, Ser at position 364, Ser at position 383, Asn at position 384, Gly at position 385, Gln at position 386, Pro at position 387, Asn at position 390, Val at position 397, and Val at position 422 (EU numbering) in the amino acid sequence of the first polypeptide and/or the second polypeptide.

16. The polypeptide of claim 15, wherein the amino acid alteration for conferring difference to isoelectric points is characterized by the introduction of a mutation to at least one amino acid selected from the group consisting of Gln at position 196, Ile at position 199, Ala at position 231, Glu at position 233, Leu at position 242, Val at position 263, Glu at position 272, Gly at position 316, Leu at position 358, Ser at position 364, Ser at position 383, Pro at position 387, and Val at position 397 (EU numbering) in the amino acid sequence of either polypeptide of the first polypeptide or the second polypeptide;

and introduction of a mutation to at least one amino acid selected from the group consisting of Gly at position 137, Gly at position 138, Thr at position 139, Lys at position 147, Ser at position 192, Leu at position 193, Tyr at position 198, Ile at position 199, Asn at position 203, Lys at position 214, Lys at position 274, Tyr at position 278, Lys at position 288, Lys at position 290, Gly at position 316, Lys at position 317, Lys at position 320, Lys at position 324, Thr at position 335, Ser at position 337, Lys at position 338, Lys at position 340, Gly at position 341, Leu at position 358, Lys at position 360, Gln at position 362, Ser at position 383, Asn at position 384, Gly at position 385, Gln at position 386, Asn at position 390, and Val at position 422 (EU numbering) in the amino acid sequence of the other polypeptide.

17. The polypeptide of claim 1, wherein the polypeptide is an antigen-binding molecule.

18. The polypeptide of claim 17, wherein the antigen-binding molecule is an antibody, a bispecific antibody, or an Fc fusion molecule such as a peptide Fc fusion protein or scaffold Fc fusion protein.

19. A pharmaceutical composition comprising the polypeptide of claim 1 and a medically acceptable carrier.

20. A method for altering the function of a polypeptide comprising an Fc region, which comprises the steps of heterodimerizing the Fc region by introducing the amino acid mutation(s) according to claim 1 into the first polypeptide and/or the second polypeptide constituting the Fc region, and introducing an amino acid mutation(s) to alter the Fc region function compared to when the Fc region forms a homodimer.

21. A method for producing a polypeptide comprising an Fc region, which comprises the steps of heterodimerizing the Fc region by introducing the amino acid mutation(s) according to claim 1 into the first polypeptide and/or the second polypeptide constituting the Fc region, and introducing an amino acid mutation(s) to alter the Fc region function compared to when the Fc region forms a homodimer.

12. (canceled)