FCgammaRIIB-SPECIFIC FC REGION VARIANT

• US 20150299296A1
• Filed: 08/23/2013
• Published: 10/22/2015
• Est. Priority Date: 08/24/2012
• Status: Active Grant

• Current Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Katada, Hitoshi, Kadono, Shojiro, Mimoto, Futa, Igawa, Tomoyuki
• Granted Patent: US 10,919,953 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• 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 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/303 Liver or Pancreas
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/72 Increased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

An objective of the present invention is to provide an Fc region variant with enhanced FcγRIIb-binding activity, and/or enhanced binding selectivity to FcγRIIb compared to FcγRIIa (type R), as compared to those of a polypeptide containing an Fc region to which an amino acid alteration(s) has not been introduced; a polypeptide which includes the Fc region variant; a pharmaceutical composition containing the polypeptide; preventing therapeutic or preventive agent for immunological inflammatory diseases that includes the pharmaceutical composition; a production method thereof; and a method of enhancing FcγRIIb-binding activity and also enhancing binding selectivity to FcγRIIb compared to FcγRIIa (type R).

It was found that a polypeptide containing an antibody Fc region variant that contains an amino acid sequence in which an amino-acid alteration at position 238 (EU numbering) is combined with other specific amino-acid alterations enhances FcγRIIb-binding activity, and/or enhances binding selectivity to FcγRIIb compared to FcγRIIa (type R).

Claims

1. An Fc region variant in which amino acid at position 238 according to EU numbering and at least one amino acid selected from those at positions 233, 234, 235, 237, 264, 265, 266, 267, 268, 269, 271, 272, 274, 296, 326, 327, 330, 331, 332, 333, 334, 355, 356, 358, 396, 409, and 419 according to EU numbering are altered to other amino acids, wherein binding activity of the variant to Fcγ

receptors [KD value for Fcγ

RIIb of a polypeptide comprising an Fc region to which an amino acid alteration(s) has not been introduced]/[KD value for Fcγ

RIIb of a polypeptide comprising the Fc region variant] has a value which is 15.0 or greater.

2. The Fc region variant of claim 1, wherein the amino acids at positions 238, 268, and 271 according to EU numbering are altered to other amino acids, and in addition, at least one amino acid selected from amino acids at positions 233, 237, 264, 267, 272, 296, 327, 330, 332, and 396 according to EU numbering are altered to other amino acids.

5. The Fc region variant of any one of claims 1 to 4, wherein the value for [KD value for Fcγ

RIIb of a polypeptide comprising an Fc region to which an amino acid alteration(s) has not been introduccd]/[KD value for Fcγ

RIIb of a polypeptide comprising an Fc region variant] is 50.0 or more.

6. The Fc region variant of any one of claims 1 to 4, wherein the value for [KD value for Fcγ

RIIb of a polypeptide comprising an Fc region to which an amino acid alteration(s) has not been introduced]/[KD value for Fcγ

RIIb of a polypeptide comprising an Fc region variant] is 100.0 or more.

7. The Fc region variant of any one of claims 1 to 6, wherein the value for [KD value for Fcγ

RIIa (type R) of a polypeptide comprising an Fc region variant]/[KD value for Fcγ

RIIb of a polypeptide comprising an Fc region variant] is 10.0 or greater.

8. The Fc region variant of any one of claims 1 to 6, wherein the value for [KD value for Fcγ

RIIa (type R) of a polypeptide comprising an Fc region variant]/[KD value for Fcγ

RIIb of a polypeptide comprising an Fc region variant] is 20.0 or greater.

9. The Fc region variant of any one of claims 1 to 8, wherein the Fc region variant comprises any one of the following set of amino acid alterations of (a) to (x):

(a) amino acid alterations at positions 238, 233, 237, 268, 271, 296, and 330 (EU numbering) of an Fc region;

(b) amino acid alterations at positions 238, 237, 268, 271, 296, and 330 (EU numbering) of an Fc region;

(c) amino acid alterations at positions 238, 233, 237, 268, 271, 296, 330, and 332 (EU numbering) of an Fc region;

(d) amino acid alterations at positions 238, 233, 237, 264, 267, 268, 271, and 330 (EU numbering) of an Fc region;

(e) amino acid alterations at positions 238, 233, 237, 267, 268, 271, 296, 330, and 332 (EU numbering) of an Fc region;

(f) amino acid alterations at positions 238, 237, 267, 268, 271, 296, 330, and 332 (EU numbering) of an Fc region;

(g) amino acid alterations at positions 238, 233, 237, 268, 271, 296, 327, and 330 (EU numbering) of an Fc region;

(h) amino acid alterations at positions 238, 233, 237, 264, 267, 268, and 271 (EU numbering) of an Fc region;

(i) amino acid alterations at positions 238, 233, 237, 264, 267, 268, 271, 296, and 330 (EU numbering) of an Fc region;

(j) amino acid alterations at positions 238, 233, 237, 264, 267, 268, 271, 296, 330, and 396 (EU numbering) of an Fc region;

(k) amino acid alterations at positions 238, 237, 264, 267, 268, 271, and 330 (EU numbering) of an Fc region;

(l) amino acid alterations at positions 238, 237, 264, 267, 268, 271, 296, and 330 (EU numbering) of an Fc region;

(m) amino acid alterations at positions 238, 264, 267, 268, and 271 (EU numbering) of an Fc region;

(n) amino acid alterations at positions 238, 264, 267, 268, 271, and 296 (EU numbering) of an Fc region;

(o) amino acid alterations at positions 238, 237, 267, 268, 271, 296, and 330 (EU numbering) of an Fc region;

(p) amino acid alterations at positions 238, 233, 237, 264, 267, 268, 271, 330, and 396 (EU numbering) of the Fc region;

(q) amino acid alterations at positions 238, 233, 237, 264, 267, 268, 271, 296, 327, 330, and 396 (EU numbering) of an Fc region;

(r) amino acid alterations at positions 238, 233, 237, 264, 267, 268, 271, 272, and 296 (EU numbering) of an Fc region;

(s) amino acid alterations at positions 238, 237, 264, 267, 268, 271, 272, and 330 (EU numbering) of an Fc region;

(t) amino acid alterations at positions 238, 237, 264, 267, 268, 271, 272, 296, and 330 (EU numbering) of an Fc region;

(u) amino acid alterations at positions 238, 233, 264, 267, 268, and 271 (EU numbering) of an Fc region;

(v) amino acid alterations at positions 238, 237, 267, 268, 271, 296, and 330 (EU numbering) of an Fc region;

(w) amino acid alterations at positions 238, 264, 267, 268, 271, 272, and 296 (EU numbering) of an Fc region; and

(x) amino acid alterations at positions 238, 233, 264, 267, 268, 271, and 296 (EU numbering) of an Fc region.

10. The Fc region variant of any one of claims 1 to 8, wherein the Fc region variant comprises any one of the following amino acid sequences of (a) to (x):

(a) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 268 is Asp, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(b) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 268 is Asp or Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(c) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 268 is Asp, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, the amino acid at position 330 is Arg, and the amino acid at position 332 is Thr, according to EU numbering, in an Fc region;

(d) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is ile, the amino acid at position 267 is Gly or Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(e) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, the amino acid at position 330 is Arg, and the amino acid at position 332 is Thr, according to EU numbering, in an Fc region;

(f) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, the amino acid at position 330 is Arg, and the amino acid at position 332 is Thr, according to EU numbering, in an Fc region;

(g) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 268 is Asp, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, the amino acid at position 327 is Gly, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(h) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Hie, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, and the amino acid at position 271 is Gly, according to EU numbering, in an Fc region;

(i) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(j) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, the amino acid at position 330 is Arg, and the amino acid at position 396 is Met or Leu, according to EU numbering, in an Fc region;

(k) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(l) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(m) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, and the amino acid at position 271 is Gly, according to EU numbering, in an Fc region;

(n) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, and the amino acid at position 296 is Asp, according to EU numbering, in an Fc region;

(o) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 267 is Ala or Gly, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(p) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 330 is Arg, and the amino acid at position 396 is Met or Leu, according to EU numbering, in an Fc region;

(q) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, the amino acid at position 327 is Gly, the amino acid at position 330 is Arg, and the amino acid at position 396 is Met, according to EU numbering, in an Fc region;

(r) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 272 is Asp, and the amino acid at position 296 is Asp, according to EU numbering, in an Fc region;

(s) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 272 is Pro, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(t) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 272 is Pro, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(u) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, and the amino acid at position 271 is Gly, according to EU numbering, in an Fc region;

(v) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 237 is Asp, the amino acid at position 267 is Gly, the amino acid at position 268 is Asp, the amino acid at position 271 is Gly, the amino acid at position 296 is Asp, and the amino acid at position 330 is Arg, according to EU numbering, in an Fc region;

(w) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, the amino acid at position 272 is Asp, and the amino acid at position 296 is Asp, according to EU numbering, in an Fc region; and

(x) an amino acid sequence in which the amino acid at position 238 is Asp, the amino acid at position 233 is Asp, the amino acid at position 264 is Ile, the amino acid at position 267 is Ala, the amino acid at position 268 is Glu, the amino acid at position 271 is Gly, and the amino acid at position 296 is Asp, according to EU numbering, in an Fc region.

12. A polypeptide comprising at least two Fc region variants of any one of claims 1 to 11, wherein the two Fc region variants are associated.

13. The polypeptide of claim 12, wherein the amino acid sequences of the two associated Fe region variants in the polypeptide are the same.

14. The polypeptide of claim 12, wherein the amino acid sequences of the two associated Fc region variants in the polypeptide are different.

15. The polypeptide of claim 14, wherein the amino acid sequences of the two associated Fe region variants have different amino acid(s) at at least one amino acid position selected from positions 235, 236, 237, 238, and 239 according to EU numbering in the Fc region variant.

16. The polypeptide of claim 15, wherein one of the amino acid sequences of the two associated Fc region variants is an amino acid sequence comprising at least one amino acid selected from Asp, Gln, Glu, or Thr at amino acid position 235, Asn at amino acid position 236, Phe or Trp at amino acid position 237, Glu, Gly, or Asn at amino acid position 238, and Asp or Glu at amino acid position 239 according to EU numbering.

17. The polypeptide of any one of claims 12 to 16, wherein the polypeptide comprising the Fc region variant is an IgG antibody.

18. The polypeptide of any one of claims 12 to 16, wherein the polypeptide comprising the Fc region variant is an Fe fusion protein molecule.

19. A pharmaceutical composition comprising the polypeptide of any one of claims 12 to 18.

3. An Fc region variant whose amino acid at position 238 according to EU numbering is Asp, and which comprises at least one amino acid selected from the amino acid group consisting of Asp at amino acid position 233, Tyr at amino acid position 234, Phe at amino acid position 235, Asp at amino acid position 237, Ile at amino acid position 264, Glu at amino acid position 265, Phe, Leu, or Met at amino acid position 266, Ala, Glu, Gly, or Gin at amino acid position 267, Asp, Gln, or Glu at amino acid position 268, Asp at amino acid position 269, Gly at amino acid position 271, Asp, Phe, Ile, Met, Asn, Pro, or Gln at amino acid position 272, Gin at amino acid position 274, Asp or Phe at amino acid position 296, Ala or Asp at amino acid position 326, Gly at amino acid position 327, Lys, Arg, or Ser at amino acid position 330, Ser at amino acid position 331, Lys, Arg, Ser, or Thr at amino acid position 332, Lys, Arg, Ser, or Thr at amino acid position 333, Arg, Ser, or Thr at amino acid position 334, Ala or Gln at amino acid position 355, Glu at amino acid position 356, Met at amino acid position 358, Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr at amino acid position 396, Arg at amino acid position 409, and Glu at amino acid position 419, wherein binding activity of the variant to Fcγ

receptors [KD value for Fcγ

RIIb of a polypeptide comprising an Fc region to which an amino acid alteration(s) has not been introduced]/[KD value for Fcγ

RIIb of a polypeptide comprising the Fc region variant] has a value which is 15.0 or greater.

4. The Fc region variant of claim 3, wherein the amino acid at position 238 is Asp, the amino acid at position 268 is Asp or Glu, and the amino acid at position 271 is Gly, according to EU numbering, and wherein the Fc region variant further comprises at least one amino acid selected from the amino acid group consisting of Asp at amino acid position 233, Asp at amino acid position 237, Ile at amino acid position 264, Ala or Gly at amino acid position 267, Asp or Pro at amino acid position 272, Asp at amino acid position 296, Gly at amino acid position 327, Arg at amino acid position330, Thr at amino acid position 332, and Leu or Met at amino acid position 396.

11. An Fc region variant consisting of any one amino acid sequence selected from among SEQ ID NOs:

43 to 68, SEQ ID NO;

70, SEQ ID NO;

71, and SEQ ID NOs;

75 to 77.