Fc REGION VARIANT
AssigneeChugai Seiyaku Kabushiki Kaisha
InventorsFuta Mimoto, Hitoshi Katada, Tomoyuki Igawa
Life of this patent
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Abstract
A polypeptide containing an antibody Fc region variant which has an amino acid sequence in which an amino acid alteration at position 238 according to EU numbering is combined with other specific amino acid alteration(s), was found to have decreased binding activities to all activating FcγRs, in particular FcγRIIa (R type), while maintaining its FcγRIIb-binding activity, when compared to a polypeptide containing a native IgG Fc region.
Claims59
An Fc region variant, wherein the variant comprises alteration of amino acid at position 238 of the Fc region according to EU numbering and any one of the amino acids of (a) to (k) below, wherein an FcγRIIb-binding activity of the variant is maintained and binding activities of the variant to all activating FcγRs are decreased when compared to those of a native IgG Fc region:
- (a) amino acid at position 235 of the Fc region according to EU numbering;
- (b) amino acid at position 237 of the Fc region according to EU numbering;
- (c) amino acid at position 241 of the Fc region according to EU numbering;
- (d) amino acid at position 268 of the Fc region according to EU numbering;
- (e) amino acid at position 295 of the Fc region according to EU numbering;
- (f) amino acid at position 296 of the Fc region according to EU numbering;
- (g) amino acid at position 298 of the Fc region according to EU numbering;
- (h) amino acid at position 323 of the Fc region according to EU numbering;
- (i) amino acid at position 324 of the Fc region according to EU numbering;
- (j) amino acid at position 330 of the Fc region according to EU numbering;
- and
- (k) at least two amino acids selected from (a) to (j).
An Fc region variant, wherein the amino acid at position 238 of the Fc region according to EU numbering is Asp, and wherein the variant comprises any one of the amino acids of (a) to (k) below, wherein an FcγRIIb-binding activity of the variant is maintained and binding activities of the variant to all activating FcγRs are decreased when compared to those of a native IgG Fc region:
- (a) Phe at amino acid position 235 of the Fc region according to EU numbering;
- (b) Gln or Asp at amino acid position 237 of the Fc region according to EU numbering;
- (c) Met or Leu at amino acid position 241 of the Fc region according to EU numbering;
- (d) Pro at amino acid position 268 of the Fc region according to EU numbering;
- (e) Met or Val at amino acid position 295 of the Fc region according to EU numbering;
- (f) Glu, His, Asn, or Asp at amino acid position 296 of the Fc region according to EU numbering;
- (g) Ala or Met at amino acid position 298 of the Fc region according to EU numbering;
- (h) Ile at amino acid position 323 of the Fc region according to EU numbering;
- (i) Asn or His at amino acid position 324 of the Fc region according to EU numbering;
- (j) His or Tyr at amino acid position 330 of the Fc region according to EU numbering;
- and
- (k) at least two amino acids selected from (a) to (j).
An Fc region variant, wherein the amino acid at position 238 of the Fc region according to EU numbering is Asp, and wherein the variant comprises the amino acids of any one of (1) to (3) below:
- (1) Met at amino acid position 241, Pro at amino acid position 268, Glu at amino acid position 296, and His at amino acid position 324 of the Fc region according to EU numbering;
- (2) Gln or Asp at amino acid position 237, Met at amino acid position 241, Glu at amino acid position 296, and His at amino acid position 330 of the Fc region according to EU numbering;
- and
- (3) Phe at amino acid position 235, Gln or Asp at amino acid position 237, Met at amino acid position 241, and Glu at amino acid position 296 of the Fc region according to EU numbering.
An Fc region variant, wherein the variant comprises alterations of amino acids at positions 238 and 271 of the Fc region according to EU numbering and any one of the amino acids of (a) to (h) below, wherein an FcγRIIb-binding activity of the variant is maintained and binding activities of the variant to all activating FcγRs are decreased when compared to those of a native IgG Fc region:
- (a) amino acid at position 234 of the Fc region according to EU numbering;
- (b) amino acid at position 235 of the Fc region according to EU numbering;
- (c) amino acid at position 236 of the Fc region according to EU numbering;
- (d) amino acid at position 237 of the Fc region according to EU numbering;
- (e) amino acid at position 239 of the Fc region according to EU numbering;
- (f) amino acid at position 265 of the Fc region according to EU numbering;
- (g) amino acid at position 267 of the Fc region according to EU numbering;
- and
- (h) amino acid at position 297 of the Fc region according to EU numbering.
An Fc region variant, wherein the amino acid at position 238 is Asp and the amino acid at position 271 is Gly in the Fc region according to EU numbering, and wherein the variant comprises any one of the amino acids of (a) to (h) below, wherein an FcγRIIb-binding activity of the variant is maintained and binding activities of the variant to all activating FcγRs are decreased when compared to those of a native IgG Fc region:
- (a) Ala, His, Asn, Lys, or Arg at amino acid position 234 of the Fc region according to EU numbering;
- (b) Ala at amino acid position 235 of the Fc region according to EU numbering;
- (c) Gln at amino acid position 236 of the Fc region according to EU numbering;
- (d) Arg or Lys at amino acid position 237 of the Fc region according to EU numbering;
- (e) Lys at amino acid position 239 of the Fc region according to EU numbering;
- (f) Lys, Asn, Arg, Ser, or Val at amino acid position 265 of the Fc region according to EU numbering;
- (g) Lys, Arg, or Tyr at amino acid position 267 of the Fc region according to EU numbering;
- and
- (h) Ala at amino acid position 297 of the Fc region according to EU numbering.
An Fc region variant, wherein the amino acid at position 238 is Asp and the amino acid at position 271 is Gly in the Fc region according to EU numbering, and wherein the variant comprises the amino acids of any one of (1) to (3) below:
- (1) Asp at amino acid position 233, Asp at amino acid position 238, Ile at amino acid position 264, Arg at amino acid position 267, Glu at amino acid position 268, and Gly at amino acid position 271 of the Fc region according to EU numbering;
- (2) Asp at amino acid position 233, Asp at amino acid position 237, Asp at amino acid position 238, Ile at amino acid position 264, Ala at amino acid position 267, Glu at amino acid position 268, Gly at amino acid position 271, Asp at amino acid position 296, Ala at amino acid position 297, Arg at amino acid position 330, and Met at amino acid position 396 of the Fc region according to EU numbering;
- and
- (3) Asp at amino acid position 233, Asp at amino acid position 238, Ile at amino acid position 264, Arg at amino acid position 267, Pro at amino acid position 268, Gly at amino acid position 271, and Glu at amino acid position 296 of the Fc region according to EU numbering.
An Fc region variant, wherein the variant comprises alterations of amino acids at positions 238, 271, 327, 330, and 331 of the Fc region according to EU numbering, wherein an FcγRIIb-binding activity of the variant is maintained and binding activities of the variant to all activating FcγRs are decreased when compared to those of a native IgG Fc region.
An Fc region variant, wherein the amino acid at position 238 is Asp, the amino acid at position 271 is Gly, the amino acid at position 327 is Gly, the amino acid at position 330 is Ser, and the amino acid at position 331 is Ser in the Fc region according to EU numbering, wherein an FcγRIIb-binding activity of the variant is maintained and binding activities of the variant to all activating FcγRs are decreased when compared to those of a native IgG Fc region.
An Fc region variant, wherein the amino acid at position 238 is Asp and the amino acid at position 271 is Gly in the Fc region according to EU numbering, wherein the variant also comprises the amino acids of any one of (1) to (4) below:
- (1) Asp at amino acid position 237, Asp at amino acid position 238, Asp or Glu at amino acid position 268, Gly at amino acid position 271, Gly at amino acid position 327, Ser at amino acid position 330, and Ser at amino acid position 331 of the Fc region according to EU numbering;
- (2) Asp at amino acid position 233, Asp at amino acid position 237, Asp at amino acid position 238, Asp at amino acid position 268, Gly at amino acid position 271, Gly at amino acid position 327, Ser at amino acid position 330, and, Ser at amino acid position 331 of the Fc region according to EU numbering;
- (3) Asp at amino acid position 238, Ala at amino acid position 267, Glu at amino acid position 268, Gly at amino acid position 271, Gly at amino acid position 327, Ser at amino acid position 330, and Ser at amino acid position 331 of the Fc region according to EU numbering;
- and
- (4) Asp at amino acid position 238, Ile at amino acid position 264, Ala at amino acid position 267, Gly at amino acid position 271, Gly at amino acid position 327, Ser at amino acid position 330, and Ser at amino acid position 331 of the Fc region according to EU numbering.
A method of reducing bindings of a polypeptide comprising an Fc region to all activating FcγRs while maintaining an FcγRIIb-binding activity of the polypeptide, which comprises altering the amino acid at position 238 of the Fc region according to EU numbering and at least one amino acid selected from the amino acids at positions 235, 237, 241, 268, 295, 296, 298, 323, 324, and 330 of the Fc region according to EU numbering to other amino acids.
A method of producing a polypeptide comprising an Fc region variant, wherein the method comprises altering the amino acid at position 238 of the Fc region according to EU numbering and at least one amino acid selected from the amino acids at positions 235, 237, 241, 268, 295, 296, 298, 323, 324, and 330 of the Fc region according to EU numbering to other amino acids, wherein bindings to all activating FcγRs are decreased while an FcγRIIb-binding activity is maintained as compared to those of the polypeptide before the altering.
A method of reducing binding activities of a polypeptide comprising an Fc region to all activating FcγRs while maintaining an FcγRIIb-binding activity at a similar level in comparison to those of a native IgG, wherein the method comprises combining and introducing an amino acid alteration(s) that increases the FcγRIIb-binding activity by two-fold or more as compared to that of a native IgG Fc region, and an amino acid alteration(s) that decreases the binding activities to all FcγRs.
A method of producing a polypeptide comprising an Fc region variant, wherein binding activities to all activating FcγRs are decreased while an FcγRIIb-binding activity is maintained at a similar level in comparison to those of a native IgG, wherein the method comprises combining and introducing an amino acid alteration(s) that increases the FcγRIIb-binding activity by two-fold or more as compared to that of a native IgG Fc region, and an amino acid alteration(s) that decreases the binding activities to all FcγRs.
A method of reducing bindings of a polypeptide comprising an Fc region to all activating FcγRs while maintaining an FcγRIIb-binding activity of the polypeptide, which comprises altering the amino acids at positions 238, 271, 327, 330, and 331 of the Fc region according to EU numbering to other amino acids, or further comprises altering at least one amino acid selected from the amino acids at positions 233, 237, 264, 267, and 268 of the Fc region according to EU numbering to another amino acid.
A method of producing a polypeptide comprising an Fc region variant, wherein the method comprises altering the amino acids at positions 238, 271, 327, 330, and 331 of the Fc region according to EU numbering to other amino acids, or further comprising altering at least one amino acid selected from the amino acids at positions 233, 237, 264, 267, and 268 of the Fc region according to EU numbering to another amino acid, wherein bindings to all activating FcγRs are decreased and complement-binding is decreased while an FcγRIIb-binding activity is maintained as compared to those of the polypeptide before the altering.
Dependencies are inferred from the claim text.