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( 12 ) United States Patent US010047166B2 (12 ) United States Patent (10 ) Patent No. : US 10 ,047 , 166 B2 Chen et al . (45 ) Date of Patent: Aug . 14 , 2018 ( 54 ) HUMANIZED ANTI- IGE ANTIBODIES THAT 6 ,037 , 453 A * 3 /2000 Jardieu .. .. CO7K 16 / 4291 530 / 387 . 3 CROSSLINK CD23 ON B LYMPHOCYTES 6 , 066 ,718 A * 5 /2000 Hardman . .. .. C07K 16 /4291 BUT DO NOT SENSITIZE MAST CELLS 435 / 326 6 , 180 ,370 B1 * 1/ 2001 Queen .. .. .. .. .. .. CO7K 16 /00 (71 ) Applicant: Academia Sinica , Taipei ( TW ) 424 / 133 . 1 6 ,685 , 939 B2 * 2 / 2004 Jardieu CO7K 16 / 00 ( 72 ) Inventors : Jiun - Bo Chen , Taipei ( TW ) ; Yu - Yu 424 / 130 . 1 Shiung , Taipei ( TW ) ; Tse - Wen Chang , 6 ,914 , 129 B2 * 7 /2005 Jardieu .. CO7K 16 / 00 Taipei ( TW ) 424 / 133 . 1 7, 022 , 500 B1 * 4 /2006 Queen .. CO7K 16 / 00 ( * ) Notice: Subject to any disclaimer, the term of this 424 / 130 . 1 patent is extended or adjusted under 35 7 ,253 , 263 B1 * 8 / 2007 Hanai . .. CO7K 16 / 18 U . S . C . 154 ( b ) by 0 days . 7 ,531 , 169 B2 * 5 /2009 Singh . .. .. CO7K424 16/ 133 /005 . 1 424 / 130 . 1 ( 21 ) Appl. No .: 15 /318 , 360 8 , 071, 097 B2 * 12 / 2011 Wu .. .. A01K 67 /0278 424 / 133 . 1 ( 22 ) PCT Filed : Jun . 16 , 2015 8 ,080 ,249 B2 * 12 / 2011 Risk .. .. .. .. .. .. CO7K 16 /4291 424 / 133 . 1 9 ,587 , 034 B2 * 3 / 2017 Chang .. .. .. .. CO7K 16 / 4291 (86 ) PCT No. : PCT/ US2015 / 035981 2002/ 0076404 A1 * 6 / 2002 Chang . .. .. .. CO7K 16 / 4291 $ 371 ( c ) ( 1 ) , 424 / 131 . 1 2004/ 0171816 A1 * 9 / 2004 Schenk A61K 47 /646 ( 2 ) Date : Dec . 13 , 2016 530 /388 . 15 2005/ 0214857 A1 * 9 /2005 Lasters . .. .. .. .. .. .. CO7K 16 / 00 (87 ) PCT Pub . No. : WO2015/ 195631 435 / 7 . 1 PCT Pub . Date : Dec . 23 , 2015 2005 /0288491 A1 * 12 /2005 Wilson . .. .. .. .. .. CO7K 16 / 1027 530 / 388 . 15 2006 /0134098 A1 * 6 /2006 Bebbington .. .. CO7K 16 /005 Prior Publication Data 424 / 130 . 1 US 2017/ 0121425 A1 May 4 , 2017 2011/ 0312505 Al * 12 / 2011 Reddy . .. CO7K 16 /065 506 / 2 Related U .S . Application Data ( 60 ) Provisional application No . 62/ 013 , 196 , filed on Jun . OTHER PUBLICATIONS 17 , 2014 . Janeway et al. , Immunobiology , 3rd edition , 1997 , Garland Pub lishing , Inc ., pp . 3 : 1 - 3 : 11. * (51 ) Int. Ci. Rudikoff et al. , Proc Natl Acad Sci US A . Mar . 1982 ; 79 ( 6 ): 1979 A61K 39 / 395 ( 2006 .01 ) 83. * CO7K 16 / 42 ( 2006 .01 ) Shiung et al ., Immunobiology . Jul. 2012 ; 217 ( 7 ) :676 - 83 . doi : CO7K 16 / 28 ( 2006 .01 ) 10 . 1016 /j .imbio .2011 .11 .006 . Epub Nov. 25, 2011. * (52 ) U . S . CI. Kipriyanov et al ., Mol Biotechnol. Jan . 2004 ; 26 ( 1 ) : 39 -60 . * CPC .. .. CO7K 16 /4291 (2013 .01 ) ; CO7K 16 /2851 ( 2013 . 01 ) ; CO7K 2317 / 24 (2013 .01 ) ; COOK * cited by examiner 2317 / 33 ( 2013 .01 ) ; COZK 2317 / 76 ( 2013 .01 ) ( 58 ) Field of Classification Search Primary Examiner - Michael Szperka None See application file for complete search history . (57 ) ABSTRACT A novel humanized anti -IgE antibody is disclosed . The References Cited antibody is capable of binding to free IgE , membrane -bound U . S . PATENT DOCUMENTS IgE on B lymphocytes , IgE bound by CD23 , but not to IgE bound by high - affinity IgE .Fc receptor on mast cells . The 5 ,530 , 101 A * 6 / 1996 Queen . .. .. .. .. CO7K 16 / 00 present invention relates to the treatment of IgE -mediated 424 / 133 . 1 5 ,585 , 089 A * 12 / 1996 Queen . CO7K 16 /00 diseases , including allergic asthma, allergic rhinitis , atopic 424 / 133 . 1 dermatitis , food allergy , chronic spontaneous ( idiopathic ) 5 ,693 , 761 A * 12 / 1997 Queen .. .. CO7K 16 /00 urticaria , chronic rhinosinusitis , systemic mastocytosis , 435 / 252 . 3 cutaneous mastocytosis , allergic bronchopulmonary asper 5 ,693 , 762 A * 12 / 1997 Queen .. CO7K 16 /00 gillosis , recurrent idiopathic angioedema , and eosinophil 5, 958 ,708 A * 9/ 1999 Hardman .. CO7K424 16 / / 133 4291 . 1 associated gastrointestinal disorder by administering the 435 / 7 .21 anti - IgE antibody of the present invention . 5 , 994 ,511 A * 11/ 1999 Lowman .. .. .. .. .. .. .. CO7K 16 / 00 530 / 387 . 3 13 Claims, 18 Drawing Sheets U . S . Patent Aug. 14 , 2018 Sheet 1 of 18 US 10 ,047 , 166 B2 50 L WWW TXTRA * KY PWL wy 90 Ek . 40 80abc 30 VMIVA ng73 1)IDNO:YRHSWFAYWGQGTLVTVSA(SEQ 2)SEQIDNO:YRHSWFAYWGQGTLVTVSS( FIG.1 TO 70 20 7 VNV TIP the 7VKV * 11)SEQIDNO:YFDYWGQGTLVTVSS(- eing * * 110 y ViaVan * * TIT 10 TITELVAL 60 mu8D6VQVQLQQSGAELAKPGASVMLSCKASGYTF'NGYWMHWVKORPGQDLEWIGY TyTAT V,1-69/JH4IIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCAR 100 510 QVOLVQSGAEVKKPGSSVKVSCKASGYTFNGYWMHWVRQAPGQGLEWIGYhu8D6V. INPTTGHTEYNOKFKDKATLTADESSNTAYIELSSLTSDDSAVYYCARQEmu8D6V, INPTTGHTEYNQKFKDKATITADESTNTAYMELSSLRSEDTAVYYCARQEhu8D6V. mu8D6V, hu8D6V, OVQLVOSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGG69/JH4V.1- JH4-69/V,1 ww 1 1 U . S . Patent Aug . 14, 2018 Sheet 2 of 18 US 10 ,047 , 166 B2 40 90 TNEDPW Voy 80 30abcd ????? LIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTW- DIQMTQSPSSLSASVGDRVTITCRASQSIS-SYLNWYQQKPGKAPKL TTT * FIG.2 TFGQGTKVEIKR(SEQIDNO:12) TFGGGTKLEIKR(SEQIDNO:3) TFGQGTKVEIKR(SEQIDNO:4) 506070 ma TFGQGTKVEIKR(SEQID 10 100107 ??? IQLTQSPSSLSASVGDRVTITCRASQSVDYDGDTYMNWYQQKPGKAPKLD * V,1-39/J 39/J,1V- DIVLTQSPASLAVSLGQRATISCKASQSVDYDGDTYMNWYHQKPGQPPKLmu8D6V V mu8D6VKLIYAASNLDSGIPARFSGSGSGTDFTLNIHPVEEEDAATYYCOQTNEDPW hu8D6 LIYAASNLDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCOQTNEDPWVhu8D6 V1-39/Jk1 mu8D6V hu8D6VK hu8D6 atent Aug 14, 2018 Sheet 3 of 18 US 10, 047, 166 B2 L-3, -1 ???????? -3 - FIG.3 (juu/buJ}*3uo?q " RARP9 R & V 4 4 4 8 4 4 -0 -1 ??{ } -0 ??? leo ooo ??? ?? ??? ??sossessssssssssssssssssss ??? ???????????????????????????????????????????????????????????????????????* -0 450 U . S . Patent Aug . 14 , 2018 Sheet 4 of 18 US 10 ,047 , 166 B2 Sustenthemother 00L gagn?sostes 9080* thanwhat OL .0L . shtimi It Abconc.(mg/ml) OL FIG.4 w DOO * * * * * * * * * 00000 90000000000000000000000000000000000000000000000 strihanyikanlamanna EOL eguretatuna ithhehellwiththewobeWhiteandthinkthatnoonewhoiswaysame SOL s malah de memasuk FOX i W WW wwww n 20L wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww a 450 OD U . S . Patent Aug . 14, 2018 Sheet 5 of 18 US 10 ,047 , 166 B2 10001000 25nM 12.5nM 6.25nM 3.12nM - - - 800800 600600 400400 Time(s) FIG.5 Koff 200200 - . 1 . Kon| 300§ 250 2004 150 š 1004 50 i 50- 2 enU . S . Patentman wasAug . 14 , 2018 Sheetwon 6 of 18 USom 10 , 047 , 166 B2 prim 10-21 10-543 Abconc.(mg/ml) FIG.6b A - - - - - - - 10-6 - - hu8D6 - -C8D6 - - 0 A AAA morum % % % % % $ Intensity Mean Geometric 10-1 10-2 TipTimp Abconc.(mg/ml) 10-6543 . 212 211 11 FIG.6a hu8D6 TITWIT -•C8D6 SOODOODOODODGODI 0 In -1 Intensity Mean Geometric atent Aug . 14 , 2018 Sheet 7 of 18 US 10 ,047 , 166 B2 wwwwww aanmamm w 5H2 Omalizumab C8D6 logolo FIG.7 $0.01ug/ml hu8D6 10 vz.01 =1 IgGBhuman ã ö 450 OD U . S . Patent Aug . 14 , 2018 Sheet 8 of 18 US 10 ,047 , 166 B2 104 TTTTTTTTTTTTTT FITC-A hu8D6 TTTTT101102103 FIG.8C T LUULUUUlulit 100 30 10 Count 104 TTTT104 * T 101102103 FITC-A FIG.8b 101102103 FITC-A C8D6 m 5H2 FIG.8e TIT op100 TITUITITUTITITIT m010° 8888 ö8 No*10 Count Count 104 104 pampanyaTT -101102103 FITC-A 101102103 FITC-A Anti-FceR1a TTTTT FIG.8a FIG.8d T Omalizumab TTTT 4 LILULUULILULILULUI 100 LLLLLLLLLL LLLLLLLLL 100 ? ? ? ?? 0-2 Count Count U . S . Patent Aug . 14, 2018 Sheet 9 of 18 US 10 ,047 , 166 B2 hu8D6 Omalizumab Trastuzumab - - wat 100 Sergey 10-2 word FIG.9 prostornosotrostoc ** kicsitHoroso OOO * * something ond * 104 # mmsungomagaganapomwarganegaraangnagpapasarpohon songs * * doen * metons * n e * S * xo ? ö ö 0. OD450 U . S . Patent Aug . 14 , 2018 Sheet 10 of 18 US 10 ,047 , 166 B2 -hu8D6 +Omalizumab Trastuzumab ?????????????????????????????????????????????????????? 102 101 FIG.10 Soooooooooooooooooooooooooooooooooooooogost th Abconc.(ug/ml) menge * * * * ???????????????????????????????????? &sua ?????????????????????????????????????????????????????? 100 gooos Intensity Mean GeometricXW U . S . Patent Aug . 14 , 2018 Sheet 11 of 18 US 10 ,047 , 166 B2 Cununu FIG.11 won100 wougeeuguegerº 055 ODNE aseajaj aseppujwesoxay - d jo % U . S . Patent Aug . 14 , 2018 Sheet 12 of 18 US 10 ,047 , 166 B2 111 + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + jw/6111000 MYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYY 1.0 10 1 Z D HER Ba Omalizumab WWWWWWWWWWWWWWWWWWWW W WWW Inwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww hu8D6 FIG.12 wwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwwww HoWAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKAKAKKUA c8D6 IgGB? human ?? 2 450 OD atent Aug . 14 , 2018 Sheet 13 of 18 US 10 ,047 , 166 B2 104 104 TTTT TI TT FITC-A FIG.13b hu8D6 TIITUTI101102103 FITC-A FIG.130 Omalizumab 100101102103 YANA TIITTIII | II | II | IITTI TTTTTTIIIIIIITTTTTTTT 100 5040 2013101 WAXAAT 60 50 3307 240Count3301 104 104 TTTIIN TIIIIII 103 INI TTTTTTT FITC-A Anti-CD23 TUTTI101102103 FIG.13a C8D6 THINIUTTA 102 FITC-AFIG.130 101 100 ITITITI oB100 : ooo 688 600 juno Count U . S . Patent Aug. 14 , 2018 Sheet 14 of 18 US 10, 047 ,166 B2 10^ TTTTTTTTTT hu8D6:lgEIC AFITC-| FIG.14c 101010110210° . .. .. ?. .. … ??? .. .. .. .. .. .. .. .?? .. … .. .. .?%XXXX .. Count 104 TTTTTTTTTTTTTTTTTT c8D6:IgEIC 10110210° FITC-A FIG.14b 5H2:lgEIC rrrrrrrrrrrr 10110210° FITC-A FIG.14e ? r IT }TTT TTTT} - 1111111111111111111111 ?%10 8???? 10 ? ????14M Count Count 10* III TTTIT 1011021010* AFITC- Omalizumab:IgEIC 10110210° FITC-A Anti-FceR1a TTTTTT FIG.14a *TT FIG.14d TTI LLLLLLLLLLLL .ek1L I 10°L? ???588 10° ????588LLLLLLLLLLLL Count
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