Optimized Fc variants and methods for their generation

• US 20060160996A9
• Filed: 03/26/2004
• Published: 07/20/2006
• Est. Priority Date: 03/01/2002
• Status: Active Grant

• Current Assignee: Xencor, Inc.
• Original Assignee: Xencor, Inc.
• Inventors: Lazar, Gregory Alan, Dang, Wei, Karki, Sher Bahadur, Desjarlais, John R., Vafa, Omid, Hayes, Robert J., Chirino, Arthur J., Doberstein, Stephen Kohl
• Granted Patent: US 7,317,091 B2
• Time in Patent Office: Days
• US Class Current: 530/387.300
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• 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 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• 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 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/2893 against CD52
• CPC Class Codes: C07K 16/2896 against molecules with a "CD"-designation, not provided for elsewhere
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/3015 Breast
• CPC Class Codes: C07K 16/32 : against translation products of oncogenes
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/52 : Constant or Fc region; Isotype
• 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 2317/732 : Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 : Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2317/77 : Internalization into the cell
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present invention relates to optimized Fc variants, methods for their generation, and antibodies and Fc fusions comprising optimized Fc variants.

Claims

1. A variant protein comprising an Fc variant of a wild-type Fc polypeptide, said Fc variant comprising at least one amino acid modification in the Fc region of said wild-type Fc polypeptide, wherein said variant protein exhibits altered binding to an Fcγ

R as compared to the wild-type Fc polypeptide.

2. A variant protein according to claim 1, wherein said Fc variant comprises at least one substitution at a position corresponding to an Fc position selected from the group consisting of:

230, 240, 244, 245, 247, 262, 263, 266, 273, 275, 299, 302, 313, 323, 325, 328, and 332.

3. A variant protein according to claim 1 wherein said altered binding is an increase in affinity of said Fc variant to said Fcγ

R.

4. A variant protein according to claim 3, wherein said Fc variant binds with greater affinity to a mouse Fcγ

R.

5. A variant protein according to claim 3, wherein said said Fcγ

R is a human Fc receptor selected from the group consisting of Fcγ

RI, Fcγ

RIIa, Fcγ

RIIb, Fcγ

RIIc, and Fcγ

RIIIa.

6. A variant protein according to claim 3, wherein said Fc variant binds with greater affinity to human Fcγ

RI and Fcγ

RIIIa, but exhibits unaltered affinity to a human receptor selected from the group consisting of Fcγ

RIIa, Fcγ

RIIb, and Fcγ

RIIc.

7. A variant protein according to claim 6, wherein said Fc variant exhibits unaltered affinity to Fcγ

RIIa, Fcγ

RIIb, and Fcγ

RIIc.

8. A variant protein according to claim 3, wherein the affinity increase for binding to one or more human Fcγ

RII'"'"'s is greater than the affinity increase for binding to human Fcγ

RI and Fγ

RIIIa.

9. A variant protein according to claim 3, wherein the affinity increase is the same for binding to Fcγ

RIIa, Fcγ

RIIb, and Fcγ

RIIc.

10. A variant protein according to claim 9, wherein the affinity increase for binding to Fcγ

RIIc is greater than the affinity increase for binding to Fcγ

RIIb.

11. A variant protein according to claim 3, wherein said Fc variant binds with greater affinity to human Fcγ

RIIa, but exhibits unaltered affinity to a human receptor selected from the group consisting of Fcγ

RI, Fcγ

RIIb, Fcγ

RIIc, and Fγ

RIIa.

12. A variant protein according to claim 11, wherein said Fc variant exhibits unaltered affinity to Fcγ

RI, Fcγ

RIIb, Fcγ

RIIc, and Fγ

RIIIa.

13. A variant protein according to any of claims 1-12, wherein said Fc variant binds with greater affinity to FcRn.

14. A variant protein according to any of claims 1-12, wherein said Fc variant binds with unaltered affinity to FcRn.

15. A variant protein according to any of claims 1-12, wherein said Fc variant binds with reduced affinity to FcRn.

16. A variant protein according to any of claims 1-12, wherein said Fc variant binds with greater affinity to C1q.

17. A variant protein according to any of claims 1-12, wherein said Fc variant binds with unaltered affinity to C1q.

18. A variant protein according to claim 1 wherein said Fc variant further comprises an engineered glycoform.

19. A variant protein according to claim 18 wherein said engineered glycoform comprises an altered level of fucosylation or bisecting oligosaccharides as compared to the parent Fc polypeptide.

20. A variant protein according to claim 18 wherein said engineered glycoform improves effector function.

24. A variant protein according to claim 2 wherein said ADCC improvement is an enhancement in potency such that the EC50 of said Fc variant is approximately 5-fold greater than that of said parent Fc polypeptide.

32. A variant protein according to claim 1 that has specificity for a target antigen selected from the group consisting of CD20, CD22, CD33, CD52, Her2/neu, EGFR, EpCAM, MUC1, GD3, CEA, CA 125, HLA-DR, TNFalpha, and VEGF.

33. A variant protein according to claim 1 or 21 comprising a modification selected from the group consisting of 239D, 239E, 239N, 239Q, 239T, 240I, 240M, 264I, 264T, 264Y, 297D, 330I, 330L, 330Y, 332D, 332E, 332N, 332Q, A231E, A231G, A231K, A231P, A231Y, A298H, A327D, A327E, A327F, A327H, A327I, A327K, A327L, A327M, A327N, A327P, A327R, A327T, A327T, A327V, A327W, A327Y, A330E, A330F, A330F, A330G, A330H, A330I, A330L, A330L/I332E, A330M, A330N, A330P, A330R, A330S, A330T, A330V, A330W, A330Y, A330Y, A330Y/I332E, D221K, D221Y, D249H, D249Q, D249Y, D265F, D265F/N297E/I332E, D265G, D265H, D265I, D265K, D265L, D265M, D265N, D265P, D265Q, D265R, D265S, D265T, D265V, D265W, D265Y, D265Y/N297D/I332E, D265Y/N297D/T299L/I332E, D270F, D270G, D270H, D270I, D270L, D270M, D270P, D270Q, D270R, D270S, D270T, D270W, D270Y, D280G, D280H, D280K, D280L, D280P, D280Q, D280W, D280Y, E233A, E233D, E233F, E233G, E233H, E233I, E233K, E233L, E233M, E233N, E233Q, E233R, E233S, E233T, E233V, E233W, E233Y, E258H, E258S, E258Y, E269F, E269G, E269H, E269I, E269K, E269L, E269M, E269N, E269P, E269R, E269S, E269T, E269V, E269W, E269Y, E272D, E272F, E272G, E272H, E272I, E272K, E272L, E272M, E272P, E272R, E272S, E272T, E272V, E272W, E272Y, E283G, E283H, E283K, E283L, E283P, E283R, E283Y, E293F, E293G, E293H, E293I, E293L, E293M, E293N, E293P, E293R, E293S, E293T, E293V, E293W, E293Y, E294F, E294G, E294H, E294I, E294K, E294L, E294M, E294N, E294P, E294R, E294S, E294T, E294V, E294W, E294Y, E318H, E318L, E318Q, E318R, E318Y, E333A, E333F, E333H, E333I, E333L, E333M, E333P, E333S, E333T, E333Y, E333Y, F241D, F241E, F241E/F243Q/V262T/V264E, F241 E/F243Q/V262T/V264E/I332E, F241 E/F243R/V262E/V264R, F241 E/F243R/V262E/V264R/I332E, F241 E/F243Y/V262T/V264R, F241E/F243Y/V262T/V264R/I332E, F241L, F241L/F243L/V262I/V264I, F241L/V262I, F241R, F241R/F243Q/V262T/V264R, F241R/F243Q/V262T/V264R/I332E, F241W, F241W/F243W, F241W/F243W/V262A/V264A, F241Y, F241Y/F243Y/V262T/V264T, F241Y/F243Y/V262T/V264T/N297D/I332E, F243E, F243L, F243L/V262I/V264W, F243L/V264I, F243Q, F243R, F243W, F243W, F243Y, F275L, F275W, F275W, G236A, G236D, G236E, G236F, G236H, G236I, G236K, G236L, G236M, G236N, G236P, G236Q, G236R, G236S, G236T, G236V, G236W, G236Y, G237D, G237E, G237F, G237H, G237I, G237K, G237L, G237M, G237N, G237P, G237Q, G237R, G237S, G237T, G237V, G237W, G237Y, G281D, G281K, G281P, G281Y, H224E, H224Y, H268D, H268E, H268F, H268G, H268I, H268K, H268L, H268M, H268P, H268Q, H268R, H268T, H268V, H268W, H285D, H285E, H285K, H285Q, H285W, H285Y, I332A, I332D, I332E, I332F, I332G, I332H, I332K, I332L, I332M, I332N, I332N, I332P, I332Q, I332R, I332S, I332T, I332V, I332W, I332Y, I332Y, I336E, I336K, I336Y, K222E, K222Y, K246D, K246E, K246H, K246Y, K274D, K274E, K274F, K274G, K274H, K274I, K274L, K274M, K274N, K274P, K274R, K274S, K274T, K274V, K274W, K274Y, K288D, K288E, K288Y, K290D, K290G, K290H, K290L, K290N, K290S, K290T, K290W, K290Y, K317E, K317Q, K320D, K320F, K320G, K320H, K320I, K320L, K320N, K320P, K320S, K320T, K320T, K320V, K320W, K320Y, K322D, K322F, K322G, K322H, K322I, K322P, K322S, K322T, K322V, K322W, K322Y, K325E, K326A, K326D, K326I, K326I, K326L, K326N, K326P, K326Q, K326S, K326T, K326T, K326W, K326Y, K334A, K334E, K334F, K334I, K334P, K334T, L234A, L234D, L234E, L234F, L234G, L234H, L234I, L234I, L234I/L235D, L234K, L234M, L234N, L234P, L234Q, L234R, L234S, L234T, L234V, L234W, L234Y, L235A, L235D, L235E, L235F, L235F, L235G, L235H, L235I, L235K, L235M, L235N, L235P, L235Q, L235R, L235S, L235T, L235V, L235W, L235Y, L328A, L328D, L328D/I332E, L328E, L328E/I332E, L328F, L328G, L328H, L328H/I332E, L328I, L328I/I332E, L328K, L328M, L328M/I332E, L328N, L328N/I332E, L328P, L328Q, L328Q, L328Q/I332E, L328R, L328S, L328T, L328T/I332E, L328V, L328V/I332E, L328W, L328Y, M428F M428L, N276D, N276E, N276F, N276G, N276H, N276I, N276L, N276M, N276P, N276R, N276S, N276T, N276V, N276W, N276Y, N276Y, N286E, N286G, N286P, N286Y, N297D, N297D/A330Y/I332E, N297D/I332E, N297D/I332E/S239D/A330L, N297D/S298A/A330Y/I332E, N297D/T299E/I332E, N297D/T299F/I332E, N297D/T299H/I332E, N297D/T299I/I332E, N297D/T299L/I332E, N297D/T299V/I332E, N297E, N297E/I332E, N297F, N297G, N297H, N297I, N297K, N297L, N297M, N297P, N297Q, N297R, N297S, N297S, N297S/I332E, N297T, N297V, N297W, N297Y, N325A, N325D, N325E, N325F, N325G, N325H, N325I, N325K, N325L, N325M, N325P, N325Q, N325Q, N325R, N325S, N325T, N325V, N325W, N325Y, P227E, P227G, P227K, P227Y, P228E, P228G, P228K, P228Y, P230A, P230A/E233D, P230A/E233D/I332E, P230E, P230G, P230Y, P232E, P232G, P232K, P232Y, P238D, P238E, P238F, P238G, P238H, P238I, P238K, P238L, P238M, P238N, P238Q, P238R, P238S, P238T, P238V, P238W, P238Y, P244H, P244H/P245A/P247V, P245A, P247G, P247V, P271A, P271D, P271E, P271 F, P271G, P271 H, P271I, P271K, P271L, P271M, P271N, P271Q, P271R, P271S, P271T, P271V, P271W, P271Y, P291D, P291E, P291G, P291H, P291I, P291Q, P291T, P329D, P329E, P329F, P329F, P329G, P329H, P329I, P329K, P329L, P329M, P329N, P329Q, P329R, P329S, P329T, P329V, P329W, P329Y, P331D, P331F, P331H, P331H, P331I, P331L, P331M, P331Q, P331R, P331T, P331V, P331W, P331Y, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, R255E, R255Y, R292D, R292E, R292T, R292Y, R301D, R301E, R301H, R301Y, S239D, S239D/A330L/I332E, S239D/A330Y/I332E, S239D/A330Y/I332E/K326E, S239D/A330Y/I332E/K326T, S239D/A330Y/I332E/L234I, S239D/A330Y/I332E/L235D, S239D/A330Y/I332E/V240I, S239D/A330Y/I332E/V264T, S239D/A330Y/I332E/V266I, S239D/D265F/N297D/I332E, S239D/D265H/N297D/I332E, S239D/D265I/N297D/I332E, S239D/D265L/N297D/I332E, S239D/D265T/N297D/I332E, S239D/D265V/N297D/I332E, S239D/D265Y/N297D/I332E, S239D/I332D, S239D/I332E, S239D/I332E/A330I, S239D/I332N, S239D/I332Q, S239D/N297D/I332E, S239D/N297D/I332E/A330Y, S239D/N297D/I332E/A330Y/F241S/F243H/V262T/V264T, S239D/N297D/I332E/K326E, S239D/N297D/I332E/L235D, S239D/S298A/I332E, S239D/V264I/A330L/I332E, S239D/V264I/I332E, S239D/V264I/S298A/I332E, S239E, S239E/D265G, S239E/D265N, S239E/D265Q, S239E/I332D, S239E/I332E, S239E/I332N, S239E/I332Q, S239E/N297D/I332E, S239E/V264I/A330Y/I332E, S239E/V264I/I332E, S239E/V264I/S298A/A330Y/I332E, S239F, S239G, S239H, S239I, S239K, S239L, S239M, S239N, S239N/A330L/I332E, S239N/A330Y/I332E, S239N/I332D, S239N/I332E, S239N/I332N, S239N/I332Q, S239N/S298A/I332E, S239P, S239Q, S239Q/I332D, S239Q/I332E, S239Q/I332N, S239Q/I332Q, S239Q/V264I/I332E, S239R, S239T, S239V, S239W, S239Y, S267D, S267E, S267F, S267H, S267I, S267K, S267L, S267L/A327S, S267M, S267N, S267P, S267Q, S267Q/A327S, S267R, S267T, S267V, S267W, S267Y, S298A, S298A/I332E, S298D, S298E, S298F, S298I, S298K, S298M, S298N, S298Q, S298R, S298V, S298W, S298Y, S304D, S304H, S304L, S304N, S304T, S324D, S324F, S324G, S324H, S324I, S324L, S324M, S324P, S324R, S324T, S324V, S324W, S324Y, S337E, S337H, S337N, T223E, T223K, T225, T225E, T225K, T225W, T250E, T250Q, T260D, T260E, T260H, T260Y, T299A, T299D, T299E, T299F, T299G, T299H, T299I, T299K, T299L, T299L, T299M, T299N, T299P, T299Q, T299R, T299S, T299V, T299W, T299Y, T335D, T335F, T335G, T335H, T335I, T335L, T335M, T335N, T335P, T335R, T335S, T335V, T335W, T335Y, V240A, V240I, V240I/V266I, V240M, V240T, V262A, V262E, V262F, V262I, V262T, V263A, V263I, V263M, V263T, V264D, V264E, V264E/N297D/I332E, V264F, V264G, V264H, V264I, V264I/A330L/I332E, V264I/A330Y/I332E, V264I/I332E, V264I/S298A/I332E, V264K, V264L, V264M, V264N, V264P, V264Q, V264R, V264S, V264T, V264W, V264Y, V266A, V266I, V266M, V266T, V266T, V273I, V273I, V282E, V282G, V282K, V282P, V282Y, V284E, V284L, V284N, V284T, V284Y, V302I, V302I, V303D, V303E, V303Y, V305E, V305T, V305Y, V323I, V323I, W313F, Y278D, Y278E, Y278G, Y278H, Y278I, Y278K, Y278L, Y278M, Y278N, Y278P, Y278Q, Y278R, Y278S, Y278T, Y278V, Y278W, Y296A, Y296D, Y296D/N297D/I332E, Y296E, Y296E/N297D/I332E, Y296G, Y296H, Y296H/N297D/I332E, Y296I, Y296K, Y296L, Y296M, Y296N, Y296N/N297D/I332E, Y296Q, Y296Q/N297D/I332E, Y296R, Y296S, Y296T, Y296T/N297D/I332E, Y296V, Y300A, Y300D, Y300E, Y300G, Y300H, Y300I, Y300K, Y300L, Y300M, Y300N, Y300P, Y300Q, Y300R, Y300S, Y300T, Y300V, and Y300W.

34. A variant protein according to claim 1 or 21 wherein said variant protein is an antibody comprising said Fc variant.

35. A variant protein according to claim 1 or 21 wherein said variant protein is an Fc fusion protein comprising said Fc variant.

36. A pharmaceutical composition comprising a variant protein according to claim 1 and a pharmaceutically acceptable carrier.

37. A method of treating a mammal in need of said treatment, comprising administering a variant protein of claim 1.

21. A variant protein comprising an Fc variant of a wild-type Fc polypeptide, said Fc variant comprising at least one amino acid modification in the Fc region of said wild-type Fc polypeptide, wherein said Fc variant modulates effector function as compared to the parent Fc polypeptide.

22. A variant protein according to claim 21, wherein said effector function is ADCC.

23. A variant protein according to claim 22, wherein said Fc variant improves ADCC in the presence of human effector cells as compared to said wild-type Fc polypeptide.

25. A variant protein according to claim 23 wherein said ADCC improvement is an enhancement in potency such that the EC50 of said Fc variant is between approximately 5-fold and 1000-fold greater than that of said parent Fc polypeptide.

26. A variant protein according to claim 23 wherein said ADCC improvement is an enhancement in efficacy such that the maximal ADCC is approximately 2-fold greater than that of said parent Fc polypeptide.

27. A variant protein according to claim 26 wherein said Fc variant improves ADCC in the presence of mouse effector cells as compared to said wild-type Fc polypeptide.

28. A variant protein according to claim 21 wherein said effector function is ADCP.

29. A variant protein according to claim 28 wherein said Fc variant improves ADCP as compared to said wild-type Fc polypeptide.

30. A variant protein according to claim 25 or 29 wherein CDC is unaffected.

31. A variant protein according to claim 25 or 29 wherein CDC is ablated.