Immunoglobulin variants outside the Fc region

• US 20050244403A1
• Filed: 03/24/2005
• Published: 11/03/2005
• Est. Priority Date: 03/24/2004
• Status: Active Grant

• Current Assignee: Xencor, Inc.
• Original Assignee: Xencor, Inc.
• Inventors: Lazar, Gregory Alan, Karki, Sher Bahadur
• Granted Patent: US 7,276,585 B2
• Time in Patent Office: Days
• US Class Current: 424/130.100
• 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 16/2893 against CD52
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/567 Framework region [FR]
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present invention relates to antibody variants outside the Fc region that alter binding affinity to one or more effector ligands, methods for their generation, and their therapeutic application.

Claims

1. An antibody variant comprising an amino acid modification outside the Fc region, wherein said modification improves affinity of the antibody for one or more effector ligands relative to the parent antibody.

2. An antibody variant according to claim 1 wherein said modification is within the constant light (CL) region of said antibody.

3. An antibody variant according to claim 2 wherein said variant comprises an amino acid modification at a CL region position selected from the group consisting of 108, 109, 110, 111, 112, 114, 116, 121, 122, 123, 124, 125, 126, 127, 128, 129, 131, 137, 138, 140, 141, 142, 143, 145, 147, 149, 150, 151, 152, 153, 154, 155, 156, 157, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 176, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 193, 195, 197, 199, 200, 202, 203, 204, 205, 206, 207, 208, 210, 211, 212, and 213.

4. An antibody variant according to claim 3 wherein said CL region is human and said modification is selected from the group consisting of R108Q, R108D, R108I, T109P, T109R, T109D, V110K, V110E, V110I, A111K, A111E, A111L, A112R, A112E, A112Y, S114D, S114K, S114I, F116T, S121D, D122S, D122R, D122Y, E123R, E123L, Q124E, L125E, L125K, K126Q, K126D, K126L, S127A, S127D, S127K, G128N, T129K, T129E, T129I, S131T, N137S, N137K, N138D, N138K, N138L, Y140K, Y140E, Y140H, P141K, P141E, R142G, R142L, R142D, E143A, E143R, E143L, K145T, K145D, K145Y, Q147A, Q147E, Q147K, K149D, K149Y, V150A, D151K, D1511, N152S, N152R, N152L, A153S, A153D, A153H, L154V, L154E, L154R, Q155K, Q155E, Q1551, S156A, S156D, S156R, G157N, N158R, N158D, N158L, S159K, S159E, S159L, Q160V, Q160K, E161K, E161L, S162T, V163T, V163K, V163E, T164Q, E165P, E165K, E165Y, Q166S, Q166E, Q166M, D167K, D167L, S168Q, S168K, S168Y, K169S, K169H, K169D, D170N, D170R, D170I, S171N, S171A, S171V, T172K, T172I, T172E, Y173K, Y173Q, Y173L, S174A, S176T, T180S, T180K, T180E, L181K, S182T, S182E, S182R, K183P, K183D, K183L, A184E, A184K, A184Y, D185Q, D185R, D1851, E187K, E187Y, K188S, K188E, K188Y, H189D, H189K, H189Y, K190R, K190E, K190L, V191S, V191E, V191R, A193S, A193E, A193K, E195Q, E195K, E195I, T197E, T197K, T197L, Q199E, Q199K, Q199Y, G200S, S202D, S202R, S202Y, S203D, S203R, S203L, P204T, V205E, V205K, T206E, T206K, T206I, K207E, K207L, K207A, S208T, S208E, S208K, N210A, N210E, N210K, R211P, R211E, R211A, G212T, G212K, G212E, E213R, and E213L, wherein numbering is according to the EU numbering scheme.

5. An antibody variant according to claim 1 wherein said modification is within the constant heavy (CH) region of said antibody.

6. An antibody variant according to claim 5 wherein said variant comprises an amino acid modification at a CH region position selected from the group consisting of 118, 119, 120, 121, 122, 124, 126, 129, 131, 132, 133, 135, 136, 137, 138, 139, 147, 148, 150, 151, 152, 153, 155, 157, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 183, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 201, 203, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 216, 217, 218, 219, 221, 222, 223, 224, and 225, wherein numbering is according to the EU numbering scheme.

7. An antibody according to claim 6 wherein said CH region is human and said modification is selected from the group consisting of A118K, A118E, A118Y, S119R, S119E, S119Y, T120R, T120E, T120I, K121E, K121Y, K121H, G122E, G122R, S124K, S124E, S124Y, F126K, F126D, A129L, A129D, S131G, S131T, S132D, S132R, S132L, K133R, K133E, K133L, T135I, T135E, T135K, S136E, S136K, S136I, G137E, G138S, G138R, G138D, T139I, T139E, T139K, K147A, K147E, D148Y, D148K, F150L, F150K, F150E, P151A, P151D, E152L, E152K, P153L, P153D, T155E, T155K, T155I, S157E, S157K, S157Y, N159K, N159D, N159L, S160K, S160E, S160Y, G161D, A162D, A162K, A162Y, L163R, T164R, T164E, T164Y, S165D, S165R, S165Y, G166D, V167A, H168L, T169E, P171G, P171H, A172K, A172L, A172E, V173T, V173D, L174E, L174K, L174Y, Q175D, Q175L, S176D, S176R, S176L, S177R, S177E, S177Y, G178D, L179K, L179Y, L179E, Y180K, Y180L, Y180E, S183T, T1871, T187K, T187E, V188I, P189D, P189G, S190I, S190K, S190E, S191D, S191R, S191Y, S192N, S192R, S192L, L193F, L193E, G194R, G194D, T195R, T195D, T195Y, Q196K, Q196D, Q196L, T197R, T197E, T197Y, Y198L, I199T, I199D, I199K, N201E, N201K, N201L, N203D, N203L, N203K, K205D, K205L, K205AP206A, P206E, S207K, S207D, N208R, N208Y, T209E, T209K, T209Y, K210L, K210E, K210Y, K210A, V211R, V211E, V211Y, D212Q, D212K, D212H, D212L, D212Y, K213N, K213E, K213H, K213L, K213Y, K213A, K214N, K214E, K214H, K214L, K214Y, K214A, E216N, E216K, E216H, E216L, E216Y, P217D, P217H, P217A, P217V, P217G, K218D, K218E, K218Q, K218T, K218H, K218L, K218Y, K218A, S219D, S219E, S219Q, S219K, S219T, S219H, S219L, S219I, S219Y, D221K, D221Y, D221E, D221N, D221Q, D221R, D221S, D221T, D221H, D221A, D221V, D221L, D221I, D221F, D221M, D221W, D221P, D221G, K222E, K222Y, K222D, K222N, K222Q, K222R, K222S, K222T, K222H, K222V, K222L, K222I, K222F, K222M, K222W, K222P, K222G, K222A, T223D, T223N, T223Q, T223R, T223S, T223H, T223A, T223V, T223L, T2231, T223F, T223M, T223Y, T223W, T223P, T223G, T223E, T223K, H224D, H224N, H224Q, H224K, H224R, H224S, H224T, H224V, H224L, H224I, H224F, H224M, H224W, H224P, H224G, H224E, H224Y, H224A, T225D, T225N, T225Q, T225R, T225S, T225H, T225A, T225V, T225L, T2251, T225F, T225M, T225Y, T225P, T225G, T225E, T225K, and T225W.

8. An antibody variant according to claim 1 wherein said modification is within the J light (JL) region of said antibody.

9. An antibody variant according to claim 8 wherein said JL region is human and said modification is selected from the group consisting of Q100P, Q100G, Q100K, K103R, K103D, K103L, E105D, E105K, E105K, E105I, E105I, I106L, and K107E, wherein numbering is according to the EU numbering scheme.

10. An antibody variant according to claim 1 wherein said modification is within the J heavy (JH) region of said antibody.

11. An antibody variant according to claim 10 wherein said JH region is human and said modification is selected from the group consisting of T110I, S112D, S113D, and S113R, wherein numbering is according to the EU numbering scheme.

12. An antibody variant according to claim 1 wherein said modification is within the hinge region of said antibody.

13. An antibody variant according to claim 12, wherein said hinge region is human and wherein said modification is selected from the group consisting of D221 K, D221Y, T223E, T223K, H224E, T225E, T225K, T225W, P227K, P227G, L234D, L234E, L234Q, L234V, L234A, L234M, L234G, L235D, L235T, L235Y, G236D, G236E, G236S, G236T, and G236A, wherein numbering is according to the EU numbering scheme.

14. An antibody variant according to claim 1 wherein said modification is within the variable light (VL) region of said antibody.

15. An antibody variant according to claim 14 wherein said VL region is from Herceptin®

and wherein said variant comprises an amino acid modification at a position selected from the group consisting of 3, 10, 22, 24, 28, 30, 31, 32, 40, 42, 43, 50, 51, 52, 53, 55, 56, 66, 83, 87, 92, 93, 94, 96, 100, 103, 105, 106, 107, and 108, wherein numbering is according to the EU numbering scheme.

16. An antibody variant according to claim 15 wherein said modification is selected from the group consisting of Q3K, S10F, T22N, R24K, D28N, N30D, T31K, A32Y, P40L, K42E, A43S, S50N, A51T, S52N, F53N, Y55Q, S56T, R66G, F83V, Y87F, Y921, T93S, T94R, P96R, Q100T, K103A, E105A, I106L, K107A, R108A, Q100P, Q100G, Q100K, K103R, K103D, K103L, E105D, E105K, E105I, I106L, K107E, and K107L.

17. An antibody variant according to claim 1 wherein said modification is within the variable heavy (VH) region of said antibody.

18. An antibody variant according to claim 17 wherein said VL region is from Herceptin®

and wherein said variant comprises an amino acid modification at a position selected from the group consisting of 3, 5, 18, 24, 28, 30, 32, 35, 40, 41, 44, 45, 50, 52, 52a, 52b, 52c, 53, 54, 55, 56, 58, 60, 61, 71, 73, 74, 75, 77, 82b, 89, 95, 97, 98, 99, 100, 100a, 100b, 102, 105, 107, 108, 110, 112, and 113, wherein numbering is according to the Kabat numbering scheme.

19. An antibody variant according to claim 1, wherein said modification is in the CH1 region.

20. An antibody variant according to claim 19, wherein said CH1 region is human and wherein said modification is selected from the group consisting of A118K, A118E, A118Y, S119E, T120E, K121H, G122E, N201E, and P206E, wherein numbering is according to the EU numbering scheme.

21. An antibody variant according to claim 1, wherein said variant binds with greater affinity to one or more FcgRs relative to said parent antibody.

22. An antibody variant according to claim 9, wherein said FcgR is human FcgRIIIa.