Fc variants with optimized properties

• US 20060235208A1
• Filed: 03/31/2006
• Published: 10/19/2006
• Est. Priority Date: 09/27/2002
• Status: Abandoned Application

• Current Assignee: Xencor, Inc.
• Original Assignee: Xencor, Inc.
• Inventors: Lazar, Gregory Alan, Dang, Wei, Karki, Sher Bahadur, Desjarlais, John R.
• Application Number:
  US11/396,495
• Publication Number:
  US 20060235208A1
• Time in Patent Office: Days
• US Class Current: 530/388.220
• 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 16/2893 against CD52
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present invention relates to Fc variants with optimized properties, methods for their generation, Fc polypeptides comprising Fc variants with optimized properties, and methods for using Fc variants with optimized properties.

Claims

1. An Fc variant of a parent Fc polypeptide, wherein said Fc variant exhibits altered binding to at least on Fcγ

R or altered antibody dependent cell-mediated cytotoxicity as compared to said parent Fc polypeptide, wherein said Fc variant comprises at least one amino acid modification in the Fc region of said parent Fc polypeptide, wherein said modification is selected from the group consisting of;

227G, 234D, 234E, 234G, 234I, 234Y, 235D, 235I, 235S, 236S, 239D, 246H, 255Y, 258H, 260H, 2641, 267D, 267E, 268D, 268E, 272H, 272I, 272R, 281D, 282G, 283H, 284E, 293R, 295E, 304T, 324G, 324I, 327D, 327A, 328A, 328D, 328E, 328F, 328I, 328M, 328N, 328Q, 328T, 328V, 328Y, 330I, 330L, 330Y, 332D, 332E, 335D, an insertion of G between positions 235 and 236, an insertion of A between positions 235 and 236, an insertion of S between positions 235 and 236, an insertion of T between positions 235 and 236, an insertion of N between positions 235 and 236, an insertion of D between positions 235 and 236, an insertion of V between positions 235 and 236, an insertion of L between positions 235 and 236, an insertion of G between positions 235 and 236, an insertion of A between positions 235 and 236, an insertion of S between positions 235 and 236, an insertion of T between positions 235 and 236, an insertion of N between positions 235 and 236, an insertion of D between positions 235 and 236, an insertion of V between positions 235 and 236, an insertion of L between positions 235 and 236, an insertion of G between positions 297 and 298, an insertion of A between positions 297 and 298, an insertion of S between positions 297 and 298, an insertion of D between positions 297 and 298, an insertion of G between positions 326 and 327, an insertion of A between positions 326 and 327, an insertion of T between positions 326 and 327, an insertion of D between positions 326 and 327, and an insertion of E between positions 326 and 327, wherein numbering is according to the EU index.

2. An Fc variant according to claim 1, wherein said variant is selected from the group consisting of:

227G/332E, 234D/332E, 234E/332E, 234Y/332E, 234I/332E, 234G/332E, 235I/332E, 235S/332E, 235D/332E, 235E/332E, 236S/332E, 236A/332E, 236S/332D, 236A/332D, 239D/268E, 246H/332E, 255Y/332E, 258H/332E, 260H/332E, 264I/332E, 267E/332E, 267D/332E, 268D/332D, 268E/332D, 268E/332E, 268D/332E, 268E/330Y, 268D/330Y, 272R/332E, 272H/332E, 283H/332E, 284E/332E, 293R/332E, 295E/332E, 304T/332E, 324I/332E, 324G/332E, 324I/332D, 324G/332D, 327D/332E, 328A/332E, 328T/332E, 328V/332E, 328I/332E, 328F/332E, 328Y/332E, 328M/332E, 328D/332E, 328E/332E, 328N/332E, 328Q/332E, 328A/332D, 328T/332D, 328V/332D, 328I/332D, 328F/332D, 328Y/332D, 328M/332D, 328D/332D, 328E/332D, 328N/332D, 328Q/332D, 330L/332E, 330Y/332E, 330I/332E, 332D/330Y, 335D/332E, 239D/332E, 239D/332E/330Y, 239D/332E/330L, 239D/332E/330I, 239D/332E/268E, 239D/332E/268D, 239D/332E/327D, 239D/332E/284E, 239D/268E/330Y, 239D/332E/268E/330Y, 239D/332E/327A, 239D/332E/268E/327A, 239D/332E/330Y/327A, 332E/330Y/268 E/327A, 239D/332E/268E/330Y/327A, Insert G>

297-298/332E, Insert A>

297-298/332E, Insert S>

297-298/332E, Insert D>

297-298/332E, Insert G>

326-327/332E, Insert A>

326-327/332E, Insert T>

326-327/332E, Insert D>

326-327/332E, Insert E>

326-327/332E, Insert G>

235-236/332E, Insert A>

235-236/332E, Insert S>

235-236/332E, Insert T>

235-236/332E, Insert N>

235-236/332E, Insert D>

235-236/332E, Insert V>

235-236/332E, Insert L>

235-236/332E, Insert G>

235-236/332D, Insert A>

235-236/332D, Insert S>

235-236/332D, Insert T>

235-236/332D, Insert N>

235-236/332D, Insert D>

235-236/332D, Insert V>

235-236/332D, and Insert L>

235-236/332D, wherein numbering is according to the EU index.

3. An Fc variant according to claim 1, wherein said Fc polypeptide composes a human IgG1 antibody.

4. An Fc variant according to claim 3, wherein said Fc polypeptide composes a human IgG1 antibody, wherein said IgG1 antibody has the allotypic residues 356D and 358L.

5. An Fc variant according to claim 3, wherein said Fc polypeptide composes a human IgG1 antibody, wherein said IgG1 antibody has the allotypic residues 356E and 358M.

6. An Fc variant according to claim 1, wherein said Fc polypeptide composes a IgG(1/2) ELLGG antibody as defined by SEQ ID NO:

12.

7. An IgG1 variant of a parent IgG1 polypeptide, wherein said IgG1 variant exhibits improved binding to at least on Fcγ

R, wherein said Fc variant comprises at least one amino acid insertion in the Fc region of said parent IgG1 polypeptide.

8. An IgG1 variant according to claim 7, wherein said insertion occurs between two positions, wherein said two positions are selected from the group consisting of 235 and 236, 297 and 298, and 326 and 327, wherein numbering is according to the EU index.

9. An Fc variant of a parent Fc polypeptide, wherein said Fc variant exhibits altered complement dependent cytotoxicity as compared to said parent Fc polypeptide, wherein said Fc variant comprises at least one amino acid modification in the Fc region of said parent Fc polypeptide, wherein said modification is selected from the group consisting of:

233I, 234Y, 235D, 235Y, 239D, 267D, 267E, 267Q, 268D, 268E, 268F, 268G, 271A, 271D, 271I, 272H, 272I, 272R, 274E, 274R, 274Y, 276D, 276L, 276S, 278E, 278H, 278Q, 278R, 281D, 282G, 284D, 284E, 284T, 285Y, 293R, 300D, 300T, 320I, 320T, 320Y, 322H, 322T, 322Y, 324D, 324H, 324I, 324L, 324T, 324V, 326L, 326T, 327A, 327D, 327H, 327R, 328Q, 330E, 330G, 330H, 330I, 330L, 330N, 330V, 330Y, 331D, 331L, 332E, 333F, 334T, 335D, and 335Y, wherein numbering is according to the EU index.

10. An Fc variant of a parent Fc polypeptide, wherein said Fc variant exhibits improved complement dependent cytotoxicity as compared to said parent Fc polypeptide, wherein said Fc variant comprises at least one amino acid modification in the Fc region of said parent Fc polypeptide, wherein said modification is at a position selected from the group consisting of:

235, 239, 267, 268, 272, 276, 278, 282, 284, 285, 293, 300, 322, 324, 327, 330, 331, 332, and 335, wherein numbering is according to the EU index.

11. An Fc variant according to claim 10, wherein said Fc variant comprises one or more amino acid modifications selected from the group consisting of:

239D, 267D, 267Q, 268D, 268E, 268F, 268G, 2721, 276D, 276L, 276S, 278R, 282G, 284T, 285Y, 293R, 300T, 324I, 324T, 324V, 327D, 330H, 330S, 332E, 335D.

12. An Fc variant of a parent Fc polypeptide, wherein said Fc variant exhibits reduced binding to one or more Fcγ

Rs, reduced antibody dependent cell-mediated cytotoxicity, or reduced complement dependent cytotoxicity as compared to said parent Fc polypeptide, wherein said Fc variant comprises at least one amino acid modification in the Fc region of said parent Fc polypeptide, wherein said modification is selected from the group consisting of;

232G, 234G, 234H, 235D, 235G, 235H, 236I, 236N, 236P, 236R, 237K, 237L, 237N, 237P, 238K, 239R, 265G, 267R, 269R, 270H, 297S, 299A, 299I, 299V, 325A, 325L, 327R, 328R, 329K, 330I, 330L, 330N, 330P, 330R, and 331L, wherein numbering is according to the EU index.

13. An Fc variant according to claim 12, wherein said Fc polypeptide comprises a variant selected from the group consisting of:

236R/237K, 236R/325L, 236R/328R, 237K/325L, 237K/328R, 325L/328R, 235G/236R, 267R/269R, 234G/235G, 236R/237K/325L, 236R/325L/328R, 235G/236R/237K, and 237K/325L/328R.

14. An Fc variant of a parent Fc polypeptide, wherein said Fc variant reduces by at least 10 fold relative to said parent Fc polypeptide:

affinity to human Fcγ

RIIIa, affinity to human Fcγ

RI, and complement dependent cytotoxicity.

15. An Fc variant according to claim 14, wherein said Fc polypeptide comprises at least one modification at a position selected from the group consisting of:

234G, 235G, 236R, 237K, 267R, 269R, 325L, 328R, wherein numbering is a according to the EU index.

16. An IgG antibody that targets CTLA-4, TNFα

, VEGF, α

4-integrin, or CD32b, wherein said antibody has reduced Fcγ

R affinity, reduced antibody dependent cell-mediated cytotoxicity, or reduced complement dependent cytotoxicity relative to a WT antibody.

17. An IgG antibody according to claim 16, wherein said antibody comprises an amino acid modification at a position selected from the group consisting of:

234, 235, 236, 237, 267, 269, 325, and 328, relative to a parent IgG antibody, wherein numbering is according to the EU index.

18. An IgG antibody according to claim 16, wherein said antibody is a human IgG2 or IgG4 antibody.

19. An IgG antibody variant of a parent IgG antibody, wherein said IgG variant exhibits improved antibody dependent cell-mediated cytotoxicity in the presence of competing IgG.