OPTIMIZED Fc VARIANTS AND METHODS FOR THEIR GENERATION

Abstract

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

1 . An antibody and/or immunoadhesin of a parent Fc polypeptide, said antibody and/or immunoadhesin comprising at least one amino acid modification in the Fc region of said parent Fc polypeptide at position 332, wherein said antibody and/or immunoadhesin protein exhibits altered binding to an FcγR as compared to the parent Fc polypeptide, wherein numbering is according to the EU index.  
     
     
         2 . An antibody and/or immunoadhesin according to  claim 1 , wherein said altered binding is an increase in affinity of said antibody and/or immunoadhesin to FcγRIIIa.  
     
     
         3 . An antibody and/or immunoadhesin according to  claim 1 , wherein said altered binding is an increase in affinity of said antibody and/or immunoadhesin to FcγRIIa.  
     
     
         4 . An antibody and/or immunoadhesin according to of  claim 1 , wherein said antibody and/or immunoadhesin further comprises an engineered glycoform.  
     
     
         5 . An antibody and/or immunoadhesin according to  claim 4  wherein said engineered glycoform comprises an altered level of fucosylation or bisecting oligosaccharides as compared to the parent Fc polypeptide.  
     
     
         6 . An antibody and/or immunoadhesin according to  claim 4  wherein said engineered glycoform improves effector function.  
     
     
         7 . An antibody and/or immunoadhesin according to  claim 6 , wherein said antibody and/or immunoadhesin effector function is selected from the group consisting of antibody dependent cell-mediated cytotoxicity (ADCC) and antibody dependent cell-mediated phagocytosis (ADCP) function as compared to the parent Fc polypeptide.  
     
     
         8 . An antibody and/or immunoadhesin according to  claim 7 , wherein said antibody and/or immunoadhesin improves ADCC in the presence of human effector cells as compared to said parent Fc polypeptide.  
     
     
         9 . An antibody and/or immunoadhesin according to  claim 8  wherein said ADCC improvement is an enhancement in potency such that the EC50 of said antibody and/or immunoadhesin is approximately 5-fold greater than that of said parent Fc polypeptide.  
     
     
         10 . An antibody and/or immunoadhesin according to  claim 8 , wherein said ADCC improvement is an enhancement in potency such that the EC50 of said antibody and/or immunoadhesin is between approximately 5-fold and 1000-fold greater than that of said parent Fc polypeptide.  
     
     
         11 . An antibody and/or immunoadhesin according to  claim 8 , 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.  
     
     
         12 . An antibody and/or immunoadhesin according to  claim 11 , wherein said antibody and/or immunoadhesin improves ADCC in the presence of mouse effector cells as compared to said parent Fc polypeptide.  
     
     
         13 . An antibody and/or immunoadhesin according to  claim 7 , wherein said effector function is ADCP and said antibody and/or immunoadhesin improves ADCP as compared to said parent Fc polypeptide.  
     
     
         14 . An antibody and/or immunoadhesin according to any of claims  1 - 13  wherein said polypeptide has specificity for a target antigen selected from the group consisting of CD4, CD19, CD20, CD22, CD25, CD30, CD33, CD52, CD80, B7-1, B7-2, CLTA-4, BAFF-R, Her2/neu, EGFR, EpCAM, MUC1, GD3, CEA, CA 125, HLA-DR, TNFalpha, and VEGF.  
     
     
         15 . An antibody and/or immunoadhesin according to any of claims  1 - 13  comprising wherein said modification is selected from the group consisting of 332A, 332D, 332E, 332F, 332G, 332H, 332K, 332L, 332M, 332N, 332P, 332Q, 332R, 332S, 332T, 332V, 332W, and 332Y.  
     
     
         16 . An antibody and/or immunoadhesin according to  claim 14  comprising wherein said modification is selected from the group consisting of 332A, 332D, 332E, 332F, 332G, 332H, 332K, 332L, 332M, 332N, 332P, 332Q, 332R, 332S, 332T, 332V, 332W, and 332Y.  
     
     
         17 . An antibody and/or immunoadhesin according to  claim 15  wherein said modification is selected from the group consisting of 332A, 332D, 332E, 332H, 332N, 332Q, 332T, and 332Y.  
     
     
         18 . An antibody and/or immunoadhesin according to any of claims  1 - 13  further comprising at least one modification selected from the group consisting of 227E, 227G, 227K, 227Y, 228E, 228G, 228K, 228Y, 230A, 230E, 230G, 230Y, 231E, 231G, 231K, 231P, 231Y, 232E, 232G, 232K, 232Y, 233A, 233D, 233F, 233G, 233H, 233I, 233K, 233L, 233M, 233N, 233Q, 233R, 233S, 233T, 233V, 233W, 233Y, 234A, 234D, 234E, 234F, 234G, 234H, 234I, 234K, 234M, 234N, 234P, 234Q, 234R, 234S, 234T, 234V, 234W, 234Y, 235A, 235D, 235E, 235F, 235G, 235H, 235I, 235K, 235M, 235N, 235P, 235Q, 235R, 235S, 235T, 235V, 235W, 235Y, 236A, 236D, 236E, 236F, 236H, 236I, 236K, 236L, 236M, 236N, 236P, 236Q, 236R, 236S, 236T, 236V, 236W, 236Y, 237D, 237E, 237F, 237H, 237I, 237K, 237L, 237M, 237N, 237P, 237Q, 237R, 237S, 237T, 237V, 237W, 237Y, 238D, 238E, 238F, 238G, 238H, 238I, 238K, 238L, 238M, 238N, 238Q, 238R, 238S, 238T, 238V, 238W, 238Y, 239D, 239E, 239F, 239G, 239H, 239I, 239K, 239L, 239M, 239N, 239P, 239Q, 239R, 239T, 239V, 239W, 239Y, 240A, 240I, 240M, 240T, 241D, 241E, 241L, 241R, 241S, 241W, 241Y, 243E, 243H, 243L, 243Q, 243R, 243W, 243Y, 244H, 245A, 246D, 246E, 246H, 246Y, 247G, 247V, 249H, 249Q, 249Y, 255E, 255Y, 258H, 258S, 258Y, 260D, 260E, 260H, 260Y, 262A, 262E, 262F, 262I, 262T, 263A, 263I, 263M, 263T, 264A, 264D, 264E, 264F, 264G, 264H, 264I, 264K, 264L, 264M, 264N, 264P, 264Q, 264R, 264S, 264T, 264W, 264Y, 265F, 265G, 265H, 265I, 265K, 265L, 265M, 265N, 265P, 265Q, 265R, 265S, 265T, 265V, 265W, 265Y, 266A, 266I, 266M, 266T, 267D, 267E, 267F, 267H, 267I, 267K, 267L, 267M, 267N, 267P, 267Q, 267R, 267T, 267V, 267W, 267Y, 268D, 268E, 268F, 268G, 268I, 268K, 268L, 268M, 268P, 268Q, 268R, 268T, 268V, 268W, 269F, 269G, 269H, 269I, 269K, 269L, 269M, 269N, 269P, 269R, 269S, 269T, 269V, 269W, 269Y, 270F, 270G, 270H, 270I, 270L, 270M, 270P, 270Q, 270R, 270S, 270T, 270W, 270Y, 271A, 271D, 271E, 271F, 271G, 271H, 271I, 271K, 271L, 271M, 271N, 271Q, 271R, 271S, 271T, 271V, 271W, 271Y, 272D, 272F, 272G, 272H, 272I, 272K, 272L, 272M, 272P, 272R, 272S, 272T, 272V, 272W, 272Y, 273I, 274D, 274E, 274F, 274G, 274H, 274I, 274L, 274M, 274N, 274P, 274R, 274S, 274T, 274V, 274W, 274Y, 275L, 275W, 276D, 276E, 276F, 276G, 276H, 276I, 276L, 276M, 276P, 276R, 276S, 276T, 276V, 276W, 276Y, 278D, 278E, 278G, 278H, 278I, 278K, 278L, 278M, 278N, 278P, 278Q, 278R, 278S, 278T, 278V, 278W, 280G, 280K, 280L, 280P, 280W, 281D, 281K, 281P, 281Y, 282E, 282G, 282K, 282P, 282Y, 283G, 283H, 283K, 283L, 283P, 283R, 283Y, 284E, 284L, 284N, 284T, 284Y, 285D, 285E, 285K, 285Q, 285W, 285Y, 286E, 286G, 286P, 286Y, 288D, 288E, 288Y, 290D, 290H, 290L, 290N, 290W, 291D, 291E, 291G, 291H, 291I, 291Q, 291T, 292D, 292E, 292T, 292Y, 293F, 293G, 293H, 293I, 293L, 293M, 293N, 293P, 293R, 293S, 293T, 293V, 293W, 293Y, 294F, 294G, 294H, 294I, 294K, 294L, 294M, 294P, 294R, 294S, 294T, 294V, 294W, 294Y, 295D, 295E, 295F, 295G, 295H, 295I, 295M, 295N, 295P, 295R, 295S, 295T, 295V, 295W, 295Y, 296A, 296D, 296E, 296G, 296H, 296I, 296K, 296L, 296M, 296N, 296Q, 296R, 296S, 296T, 296V, 297D, 297E, 297F, 297G, 297H, 297I, 297K, 297L, 297M, 297P, 297Q, 297R, 297S, 297T, 297V, 297W, 297Y, 298A, 298D, 298E, 298F, 298H, 298I, 298K, 298M, 298N, 298Q, 298R, 298T, 298W, 298Y, 299A, 299D, 299E, 299F, 299G, 299H, 299I, 299K, 299L, 299M, 299N, 299P, 299Q, 299R, 299S, 299V, 299W, 299Y, 300A, 300D, 300E, 300G, 300H, 300K, 300M, 300N, 300P, 300Q, 300R, 300S, 300T, 300V, 300W, 301D, 301E, 301H, 301Y, 302I, 303D, 303E, 303Y, 304D, 304H, 304L, 304N, 304T, 305E, 305T, 305Y, 313F, 317E, 317Q, 318H, 318L, 318Q, 318R, 318Y, 320D, 320F, 320G, 320H, 320I, 320L, 320N, 320P, 320S, 320T, 320V, 320W, 320Y, 322D, 322F, 322G, 322H, 322I, 322P, 322S, 322T, 322V, 322W, 322Y, 323I, 324D, 324F, 324G, 324H, 324I, 324L, 324M, 324P, 324R, 324T, 324V, 324W, 324Y, 325A, 325D, 325E, 325F, 325G, 325H, 325I, 325K, 325L, 325M, 325P, 325Q, 325R, 325S, 325T, 325V, 325W, 325Y, 326E, 326I, 326L, 326P, 326T, 327D, 327E, 327F, 327H, 327I, 327K, 327L, 327M, 327N, 327P, 327R, 327S, 327T, 327V, 327W, 327Y, 328A, 328D, 328E, 328F, 328G, 328H, 328I, 328K, 328M, 328N, 328P, 328Q, 328R, 328S, 328T, 328V, 328W, 328Y, 329D, 329E, 329F, 329G, 329H, 329I, 329K, 329L, 329M, 329N, 329Q, 329R, 329S, 329T, 329V, 329W, 329Y, 330E, 330F, 330G, 330H, 330I, 330L, 330M, 330N, 330P, 330R, 330S, 330T, 330V, 330W, 330Y, 331D, 331F, 331H, 331I, 331L, 331M, 331Q, 331R, 331T, 331V, 331W, 331Y, 333A, 333F, 333H, 333I, 333L, 333M, 333P, 333T, 333Y, 334A, 334F, 334I, 334P, 334T, 335D, 335F, 335G, 335H, 335I, 335L, 335M, 335N, 335P, 335R, 335S, 335V, 335W, 335Y, 336E, 336K, 336Y, 337E, 337H, 337N, wherein numbering is according to the EU index.  
     
     
         19 . An antibody and/or immunoadhesin according to any of claims  1 - 13  wherein said Fc region comprises modifications selected from the group consisting of 239D/332D, 239D/332E, 239D/332N, 239D/332Q, 239E/332D, 239E/332E, 239E/332N, 239E/332Q, 239N/332D, 239N/332E, 239N/332N, 239N/332Q, 239Q/332D, 239Q/332E, 239Q/332N, 239Q/332Q, 264I/332E, 297D/332E, 297E/332E, 297S/332E, 298A/332E, 328D/332E, 328E/332E, 328H/332E, 328I/332E, 328I/332E, 328M/332E, 328N/332E, 328Q/332E, 328Q/332E, 328T/332E, 328V/332E, 330L/332E, 330Y/332E, 230A/233D/332E, 239D/264I/332E, 239D/297D/332E, 239D/298A/332E, 239D/330L/332E, 239D/330Y/332E, 239D/330I/332E, 239E/264I/332E, 239E/297D/332E, 239N/298A/332E, 239N/330L/332E, 239N/330Y/332E, 239Q/264I/332E, 264E/297D/332E, 264I/298A/332E, 264I/330L/332E, 264I/330Y/332E, 265F/297E/332E, 265Y/297D/332E, 296D/297D/332E, 296E/297D/332E, 296H/297D/332E, 296N/297D/332E, 296Q/297D/332E, 296T/297D/332E, 297D/299E/332E, 297D/299F/332E, 297D/299H/332E, 297D/299I/332E, 297D/299L/332E, 297D/299I/332E, 297D/330Y/332E, 239D/264I/298A/332E, 239D/264I/330L/332E, 239D/265F/297D/332E, 239D/265H/297D/332E, 239D/265I/297D/332E, 239D/265L/297D/332E, 239D/265T/297D/332E, 239D/265V/297D/332E, 239D/265Y/297D/332E, 239D/297D/332E/235D, 239D/297D/332E/326E, 239D/297D/332E/330Y, 239D/330Y/332E/234I, 239D/330Y/332E/235D, 239D/330Y/332E/240I, 239D/330Y/332E/264T, 239D/330Y/332E/266I, 239D/330Y/332E/326E, 239D/330Y/332E/326T, 239E/264I/330Y/332E, 265Y/297D/299L/332E, 297D/298A/330Y/332E, 297D/332E/239D/330L, 239E/264I/298A/330Y/332E, 241E/243Q/262T/264E/332E, 241E/243R/262E/264R/332E, 241E/243Y/262T/264R/332E, 241R/243Q/262T/264R/332E, 241Y/243Y/262T/264T/297D/332E, and 239D/297D/332E/330Y/241S/243H/262T/264T.  
     
     
         20 . An antibody and/or immunoadhesin according to any of claims  1 - 13  wherein said antibody is a full length antibody.  
     
     
         21 . An antibody and/or immunoadhesin according to  claim 14  wherein said antibody is a full length antibody.  
     
     
         22 . An antibody and/or immunoadhesin according to  claim 21  wherein said antibody comprises a human antibody.  
     
     
         23 . An antibody and/or immunoadhesin according to  claim 1  wherein said antibody comprises a human IgG1, IgG2, IgG3, or IgG4 Fc region.  
     
     
         24 . An antibody and/or immunoadhesin according to  claim 1  wherein said antibody is an antibody fragment.  
     
     
         25 . A pharmaceutical composition comprising an antibody and/or immunoadhesin according to any of claims  1 - 13  and a pharmaceutically acceptable carrier.  
     
     
         26 . A pharmaceutical composition comprising an antibody and/or immunoadhesin according to  claim 14  and a pharmaceutically acceptable carrier.  
     
     
         27 . A pharmaceutical composition comprising an antibody and/or immunoadhesin according to  claim 18  and a pharmaceutically acceptable carrier.  
     
     
         28 . A pharmaceutical composition comprising an antibody and/or immunoadhesin according to  claim 19  and a pharmaceutically acceptable carrier.  
     
     
         29 . A method of treating a mammal in need of said treatment, comprising administering a antibody and/or immunoadhesin protein of any of claims  1 - 13 .  
     
     
         30 . A method of treating a mammal in need of said treatment, comprising administering a antibody and/or immunoadhesin protein of  claim 14 .  
     
     
         31 . A method of treating a mammal in need of said treatment, comprising administering a antibody and/or immunoadhesin protein of  claim 18 .  
     
     
         32 . A method of treating a mammal in need of said treatment, comprising administering a antibody and/or immunoadhesin protein of  claim 19.