US 20100129357A1 US 20100129357A1 (19) United States (12) Patent Application Publication oo) Pub. No.: US 2010/0129357 Al Garcia-Martinez et al. (43) Pub. Date: May 27,2010 (54) ANTIBODIES TO IL-6 AND USE THEREOF filed on Nov. 25, 2008, provisional application No. 61/117,811, filed on Nov. 25, 2008. (76) Inventors: Leon Garcia-Martinez, Woodinville, WA (US); Ann Elisabeth Carvalho Jensen, Publication Classification Edmonds, WA (US); Katie Olson, (51) Int. Cl. Kenmore, WA (US); Ben Dutzar, A61K 39/395 (2006.01) Seattle, WA (US); Ethan Ojala, A61P 37/02 (2006.01) Snohomish, WA (US); Brian Kovacevich, Snohomish, WA (US); (52) U.S. Cl.......................................................... 424/133.1 John Latham, Seattle, WA (US); Jeffrey T. L. Smith, Redmond, WA (US) (57) ABSTRACT Correspondence Address: The present invention is directed to therapeutic methods HUNTON & WILLIAMS LLP using IL-6 antagonists such as an Abl antibody or antibody INTELLECTUAL PROPERTY DEPARTMENT fragment having binding specificity for IL-6 to prevent or 1900 K STREET, N.W., SUITE 1200 treat disease or to improve survivability or quality of life of a WASHINGTON, DC 20006-1109 (US) patient in need thereof. In preferred embodiments these patients will comprise those exhibiting (or at risk of develop­ (21) Appl. No.: 12/502,581 ing) an elevated serum C-reactive protein level, reduced (22) Filed: Jul. 14, 2009 serum albumin level, elevated D-dimer or other cogulation cascade related protein(s), cachexia, fever, weakness and/or Related U.S. Application Data fatigue prior to treatment. The subject therapies also may (60) Provisional application No. 61/117,861, filed on Nov. include the administration of other actives such as chemo- 25, 2008, provisional application No. 61/117,839, therapeutics, anti-coagulants, statins, and others. AB1LOAD I AB PANEL LOAD TIME (SECONDS) PatentApplicationPublication May 27, 2010 Sheet I of 44 US 2010/0129357 Al FIG. 1 :t------ ti" AB1LOAD I AB PANEL LOAD TIME (SECONDS) PatentApplicationPublication FIG. 2 FRl CDRl FR2 CDR2 FR3 I 23 24 34 35 49 50 56 57 88 RbtVL AYDMTQTPASVEVAVGGTVTINC QASETIYSWLS WYQQKPGQPPKLLIY QASDLAS GVPSRFSGSGAGTEYTLTISGVQCDDAATYYC III I Il III I Il I Il lllllll HIM Il I I MMIMII III Mill I M Mill L12A DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY HASSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC Vl DIQMTQSPSTLSASVGDRVTITC RASQSISSWLA WYQQKPGKAPKLLIY DASSLES GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC VxO2 DIQMTQSPSSLSASVGDRVTITC RASQSISSYLN WYQQKPGKAPKLLIY AASSLQS GVPSRFSGSGSGTDFTLTISSLQPEDPATYYC VLh DIQMTQSPSTLSASVGDRVTITC QASETIYSWLS WYQQKPGKAPKLLIY QASDLAS GVPSRFSGSGSGTEFTLTISSLQPDDFATYYC CDR3 FR4 May 89 100 101 111 RbtVL QQGYSGSNVDNV FGGGTEVWKR Mill I M 27, FGGGTKVEIKR VLh FGGGTKVEIKR 2010 Sheet FRl CDRl FR2 CDR2 FR3 I 30 31 35 36 49 50 66 67 98 2 RbtVH QEQLKESGGRLVTPGTPLTLTCTASGFSLN DHAMG WVRQAPGKGLEYIG FINS-GGSARYASWAEG RFTISRTST- -TVDLKMTSLTTEDTATYFCVR of I M MM MM III Mil M 111111111MI I I III M I MMM I I Ml MM I I I 44 3-64-04 QVQLVESGGGLVQPGGSLRLSCSASGFTFS SYAMH WVRQAPGKGLEYVS AISSNGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR 3-66-04 EVQLVESGGGLVQPGGSLRLSCAASGFTVS SNYMS WVRQAPGKGLEWVS VIYS-GGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR 3-53-02 EVQLVETGGGLIQPGGSLRLSCAASGFTVS SNYMS WVRQAPGKGLEWVS VIYS-GGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR VHh QVQLVESGGGLVQPGGSLRLSCSASGFSLN DHAMG WVRQAPGKGLEYVG FINS-GGSARYAS|i|AEG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR US 2010/0129357 CDR3 FR4 99 HO 111 121 RbtVH GGAVWSIHSFDP WGPGTLVTVSS M MMMM WGQGTLVTVSS VHh GGAVWSIHSFDP WGQGTLVTVSS Al Patent Application Publication May 27, 2010 Sheet 3 of 44 US 2010/0129357 Al FIG. 3 460 123456789 IO 11 12 Π1Ι Just IgG Mff IL-Gor B-IL-G HjlHlgG and IL-G or B-IL-G I 2 3 4 5 6 7 8 9 1011 12 A B C HIIIIIH D Eilium F G H 462 I 2 3 4 5 6 7 8 9 10 11 12 A IIIIIHII B C D E F G H 463 I 2 3 4 5 6 7 8 9 10 1112 A I lllllll B Hill C D IIIIIHII E F ilium G H PatentApplicationPublication May 27, 2010 Sheet 4 of 44 US 2010/0129357 Al FIG. 4 2000-1 0.03mg/kg Ab 1 1500- Control £ 1000- 0.1 mg/kg Ab1 ID50 = 0.09273 mg/kg r2 = 0.9701 1 mg/kg Ab1 Log DoseAbI PatentApplicationPublication May 27, 2010 Sheet 5 of 44 US 2010/0129357 Al FIG. 5 100 90 Polyclonal IgG (n=9) -·- PBS (n=10) Ab 1 (n=30) 80 70 60 I I 50 O a> a. 40 30 I k 20 Ϊ-Γ4 10 O 0 5 10 15 20 25 30 Time (days) Patent Application Publication May 27, 2010 Sheet 6 of 44 US 2010/0129357 Al FIG. 6 100 m */- Polyclonal IgG (n=9) 90 ••PBS (n=10) *-Ab1 (n=30) 80 70 60 ! 50 υ <D CL 40 30 I k 20 10 O O 10 15 20 25 30 Time (days) Patent Application Publication May 27, 2010 Sheet 7 of 44 US 2010/0129357 Al FIG. 7 100 90 I I 80 1-1 Polyclonal IgG (270-320mg TW) Ab1 (270-320mg TW) I 70 k I Tc 60 > > I 50 § ω CL 40 30 20 I 10 I------A OH------ 1 I I----- —I·.· I-------III11 O 5 10 15 20 25 30 35 40 45 50 Time (days) Patent Application Publication May 27, 2010 Sheet 8 of 44 US 2010/0129357 Al FIG. 8 Polyclonal IgG (400-527mg TW) Ab1 (400-527mg TW) 0. 40- Time (days) DS 2010/0129357 Al May Sheet 9 of 44 Patent Application Publication FIG. 9 300 Time (h) PatentApplicationPublication May 27, 2010 Sheet 10 of 44 US 2010/0129357 Al FIG. 10A Ab4 BLOCKS GP130 BINDING 2.0000- ANTI BODY 1.5000-- 1.0000-- BUFFER CONTROL 0.5000-- 1500 2000 2500 3000 3500 4000 TIME (SECONDS) PatentApplication Publication May 27, 2010 Sheet 11 of 44 US 2010/0129357 Al FIG. 10B Ab3 BLOCKS GP130 BINDING 2.5000- - ANTIBODY GP130. 2.0000-- 1 1.5000-- BUFFER CONTROL 1.0000- 0.5000— 1500 2000 2500 3000 3500 TIME (SECONDS) PatentApplicationPublication May 27, 2010 Sheet 12 of 44 US 2010/0129357 Al FIG. 10C Ab8 BLOCKS GP130 BINDING -CONTROL ANTIBODY -Ab8 TIME (SECONDS) PatentApplicationPublication May 27, 2010 Sheet 13 of 44 US 2010/0129357 Al FIG. 10D Ab2 INHIBITS GP130 BINDING CONTROL ANTIBODY Ab2 8888502 M. 0.8000 CD Q m 0,6000 0.4000-- 0.2000-- 0.0000 I I I I I I > I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I 200 400 600 800 1000 1200 1400 1600 1800 TIME (SECONDS) Patent Application Publication May 27, 2010 Sheet 14 of 44 US 2010/0129357 Al FIG. 10E 1.4000-- C0NTR0L ANTIBODY 1.2000-- 1.0000-- 0.4000 — ANTIBODY 0.2000—1 200 400 600 800 1000 1200 1400 1600 1800 TIME (SECONDS) PatentApplicationPublication May 27, 2010 Sheet 15 of 44 US 2010/0129357 Al FIG. 11 Antibody Blocks IL6 Blocks IL6 Reference binding to FM binding to GP130 Ab1 Yes Yes 021051 Ab2 No Partial 021050 Ab3 No Yes 021030 Ab4 No Yes 021050 Ab6 Yes Yes 021051 Ab7 Yes Yes 021051 Ab8 No Yes 021051 PatentApplicationPublication FIG. 12 I VPPGEDSKDVAAPHR (SEQ ID NO: 590) 20 ENNLNL P KMAE KDGC (SEQ ID NO: 609) 39 QMSTKVLIQFLQKKA (SEQ ID NO: 628) 2 GEDSKDVAAPHRQPL (SEQ ID NO: 591) 21 LNLPKMAE KDGCFQS (SEQ ID NO: 610) 40 TKVLIQFLQKKAKNL (SEQ ID NO: 629) 3 SKDVAAPHRQPLTSS (SEQ ID NO: 592) 22 PKMAEKDGCFQSGFN (SEQ ID NO: 611) 41 LIQFLQKKAKNLDAI (SEQ ID NO: 630) 4 VAAPHRQPLTSSERI (SEQ ID NO: 593) 23 AEKDGCFQSGFNEET (SEQ ID NO: 612) 42 FLQKKAKNLDAITTP (SEQ ID NO: 631) 5 PHRQPLTSSERIDKQ (SEQ ID NO: 594) ■24 DGCFQSGFNEETCLV (SEQ ID NO: 613) 43 KKAKNLDAITTPDPT (SEQ ID NO: 632) 6 QPLTSSERIDKQIRY (SEQ ID NO: 595) 25 FQSGFNEETCLVKII (SEQ ID NO: 614) 44 KNLDAITTPDPTTNA (SEQ ID NO: 633) May 7 TSSERIDKQIRYILD (SEQ ID NO: 596) 26 GFNEETCLVKIITGL (SEQ ID NO: 615) 45 DAITTPDPTTNASLL (SEQ ID NO: 634) 27, ERIDKQIRYILDGIS (SEQ ID 597) 27 EETCLVKIITGLLEF (SEQ ID NO: 616) 46 TTPDPTTNASLLTKL (SEQ ID NO: 8 NO: 635) 2010 9 DKQIRYILDGISALR (SEQ ID NO: 598) 28 CLVKIITGLLEFEVY (SEQ ID NO: 617) 47 DPTTNASLLTKLQAQ (SEQ ID NO: 636) 10 IRYILDGISALRKET (SEQ ID NO: 599) 29 KIITGLLEFEVYLEY (SEQ ID NO: 618) 48 TNASLLTKLQAQNQW (SEQ ID NO: 637) Sheet 11 ILDGISALRKETCNK (SEQ ID NO: 600) 30 TGLLEFEVYLEYLQN (SEQ ID NO: 619) 49 SLLTKLQAQNQWLQD (SEQ ID NO: 638) 12 GISALRKETCNKSNM (SEQ ID NO: 601) 31 LE FEVYLEYLQNRFE (SEQ ID NO: 620) 50 TKLQAQNQWLQDMTT (SEQ ID NO: 639) 16 ■13 ALRKETCNKSNMCES (SEQ ID NO: 602) 32 EVYLEYLQNRFESSE (SEQ ID NO: 621) 51 QAQNQWLQDMTTHLI (SEQ ID NO: 640) of 14 KETCNKSNMCESSKE (SEQ ID NO: 603) 33 LEYLQNRFESSEEQA (SEQ ID NO: 622) 52 NQWLQDMTTHLILRS (SEQ ID NO: 641) 44 15 CNKSNMCE S S KEALA (SEQ ID NO: 604) 34 LQNRFESSEEQARAV (SEQ ID NO: 623) 53 LQDMTTHLILRSFKE (SEQ ID NO: 642) US 16 SNMCESSKEALAENN (SEQ ID NO: 605) 35 RFESSEEQARAVQMS (SEQ ID NO: 624) 54 MTTHLILRSFKEFLQ (SEQ ID NO: 643) 17 CESSKEALAENNLNL (SEQ ID NO: 606) 36 SSEEQARAVQMSTKV (SEQ ID NO: 625) 55 HLILRSFKEFLQSSL (SEQ ID NO: 644) 2010/0129357 18 SKEALAENNLNLPKM (SEQ ID NO: 607) 37 EQARAVQMSTKVLIQ (SEQ ID NO: 626) 56 LRSFKEFLQSSLRAL (SEQ ID NO: 645) 19 ALAENNLNLPKMAEK (SEQ ID NO: 608) 38 RAVQMSTKVLIQFLQ (SEQ ID NO: 627) ■57 FKEFLQSSLRALRQM (SEQ ID NO: 646) Al PatentApplicationPublication May 27, 2010 Sheet 17 of 44 US 2010/0129357 Al FIG. 13 PatentApplicationPublication May 27, 2010 Sheet 18 of 44 US 2010/0129357 Al FIG. 14 PatentApplicationPublication FIG. 15 A. Surface Plasmon Resonance: Averaged binding constants determined at 25 ° C for Abl to IL-6. Species (IL-6) Ka (M-1S'1) Kd (s-1) Kd Rat 1.6e6 2.2e-3 1.4 nM Mouse Lle6 4.0e-4 0.4 nM Dog Below LOQa Below LOQa Below LOQa Human 1.6e5 5e-7 4 pM May Cynomolgus 27, monkey 9.6e4 3e-6 31 pM 2010 a.