US 201401 12915A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2014/0112915 A1 Bardroff et al. (43) Pub. Date: Apr. 24, 2014 (54) IL-18 BINDING MOLECULES Publication Classification (71) Applicants: Michael Otto Bardroff, Loerrach (DE); (51) Int. Cl. Barbara Brannetti, Basel (CH); Emma C07K 6/24 (2006.01) Michelle Campbell, Surrey (GB); Beate GOIN33/68 (2006.01) Diefenbach-Streiber, Windach (DE): (52) U.S. Cl. Adina Eberth, Munich (DE); Christian CPC .......... C07K 16/244 (2013.01); G0IN33/6854 Carsten Silvester Kunz, Muenchen (2013.01) (DE); Sylwia Marshall, Epsom (GB); USPC ... 424/133.1; 424/139.1; 435/7.92; 435/69.6; Jean-Michel Rene Rondeau, Rixheim 435/331; 435/320.1; 530/387.3; 530/387.9; (FR); Jean-Marc Alfred Schlaeppi, 536/2353 Allschwil (CH); Gino Anselmus Van Heeke, Upper Beeding (GB) (57) ABSTRACT (72) Inventors: Michael Otto Bardroff, Loerrach (DE); Barbara Brannetti, Basel (CH); Emma Michelle Campbell, Surrey (GB); Beate IL-18 participates in both innate and acquired immunity. The Diefenbach-Streiber, Windach (DE): bioactivity of IL-18 is negatively regulated by the IL-18 bind Adina Eberth, Munich (DE); Christian ing protein (IL18BP), a naturally occurring and highly spe Carsten Silvester Kunz, Muenchen cific inhibitor. This soluble proteinforms a complex with free (DE); Sylwia Marshall, Epsom (GB); IL-18 preventing its interaction with the IL-18 receptor, thus Jean-Michel Rene Rondeau, Rixheim neutralizing and inhibiting its biological activity. The present (FR); Jean-Marc Alfred Schlaeppi, invention discloses binding molecules, in particular antibod Allschwil (CH); Gino Anselmus Van ies or fragments thereof, which bind IL-18 and do not bind Heeke, Upper Beeding (GB) IL-18 bound to IL-18BP (IL-18/IL-18BP complex). Apart from its physiological role, IL-18 has been shown to mediate (21) Appl. No.: 14/017,561 a variety of autoimmune and inflammatory diseases. The (22) Filed: Sep. 4, 2013 binding molecules of the inventions may be used as therapeu tic molecules for treating IL-18-related autoimmune and Related U.S. Application Data inflammatory diseases or as diagnostic tools for characteriz (60) Provisional application No. 61/697.981, filed on Sep. ing, detecting and/or measuring IL-18 not bound to IL-18BP 7, 2012. as component of the total IL-18 pool. Patent Application Publication Apr. 24, 2014 Sheet 3 of 38 US 2014/01 12915 A1 Figure 3(A) 90 800 700 a 600 s S. 500 S. ntO. 400 sO 2 300 a 200 c 100 is "hi-18BPa-FC s' hi-38BP3-Fc. 0.01 0.1 10 100 000 0.01 0.1 1 1 0 100 1000 MAb 944 (nM) MAb 94.64 (nM) i 2 : e hi-18BPa-Fc h3-18BP3-Fo 0.01 0.1 1 10 100 1000 0.01 01 000000 MAb 9465 (nM) MAb 9466 (nM) 4000 3000 5. 2000 wer f f e F hi-183P-FC hi-183Pa-Fc 0.01 0.1 10 100 1000 0.01 0.1 1 10 100 1000 MOR10579 (niM) MOR10222 (nM) Patent Application Publication Apr. 24, 2014 Sheet 4 of 38 US 2014/O112915 A1 Figure 3(B) 1400 1200 1000 e 800 600 400 200 18BPal O 1 1E-3 001 0.1 1 10 MOR08775 (nW) 1200 1000 1600 1400 e 800 1200 3 600 E 1000 - R 800 fs 400 S. 600 200 400 200 O a -1. - - - . --- . -- . .1. ----- ...............m. E4 E-3 OO1 0 1 0 100 0.0 0,1 1 0 100 1000 $888 88: 3888 . 88: 838: $288 $888 e 888 888 8:8 x 88. & 888 : 888 888 28. 88: 8.33:8: 3: ... ::::::::::::::::::: 88: 83.8 : 38 ::::::::8: & 88883 :8: 8:8::::::: Patent Application Publication Apr. 24, 2014 Sheet 5 of 38 US 2014/0112915 A1 Figure 3(C) 1200 1000 800 600 400 0 1E-4 1-3 0.01 0.1 1 10 100 18-4 18-3 0.01 0.1 1 10 100 MOR1334 (nM) MOR13342 (nM) 1200 1200 1000 1000 T 200 O O 1E-4 E-3 0.01 01 10 00 1E-4 1-3 0.01 0,1 1 0 00 MOR13347 (nM) MOR13361 (nM) 1200 1000 800 200 - 8BPa-FC 1E-4 1-3 0.01 0.1 1 10 100 MOR13363 (nM) Patent Application Publication Apr. 24, 2014 Sheet 6 of 38 US 2014/O112915 A1 Figure 3(D) 1000 800 600 400 .-188Pa-Fo E-5 E-4 1E-3 0.01 0.1 1 10 100 MORO9464 N3OK (nM) I I 1E-4 1E-3 00 0.1 1 0 100 000 MOR10222 N30S M54 (nM) Patent Application Publication Apr. 24, 2014 Sheet 7 of 38 US 2014/O112915 A1 Figure 4(A) 2000 0.01 0.1 10 100 1000 0.0 0 1 1 10 100 000 MAb 10497 (nM) MAb 10501 (nM) 8i 10 100 MA) 94.65 nM NAb 94.66 nM & & s: & 88: 88: Patent Application Publication Apr. 24, 2014 Sheet 8 of 38 US 2014/O112915 A1 Figure 4(B) 6000 5000. " is 4000 3. s" . 9 3000 e f 2000 1000 0.01 0, 10 100 000 into MA 944 nM 0.01 O. O 100 000 NAb 94.64 nM 6000 5000 E 4000 9 9 so is 2000 1000 0.01 0.1 10 100 1000 0.01 0 1 0 100 1000 MA 10579 nM MAb 10222 (nM) $8 . 3888 388 28 3888 888 x 888 8 * x 88: 8: : : 3: 388 83.3888: 8:8 Patent Application Publication Apr. 24, 2014 Sheet 9 of 38 US 2014/O112915 A1 Figure 5(A) 6000 6000 5000 500 era. 4000 A000 s s g 3000 3000 3. s 2000 f 2000 s 1000 1000 O - h8BPFC - 1889-FC O.O. O.1 10 00 000 0.01 0,1 1 10 100 1000 MOR09464 (nM) MOR9441 (nW) O, O, O 100 000 MOR10579 (nm) 5000 3000 400 250 es 20000 3000 3. 3 15000 3. 2000 e ECO 10 O O1 0,1 1 0 100 1000 1 O 100 hi-18BPa-Fc (nM) MOR08775 (nM) Patent Application Publication Apr. 24, 2014 Sheet 10 of 38 US 2014/O112915 A1 Figure 5(B) 1 10 100 MOR08776 (nM) 1000 O hL8BPa-FC 1E-4 1E-3 O.O. O.1 1 10 100 MORO9464 N3OK (nM) vs 1nM humani 18 hl 18BPa-FC E-3 0.01 0.1 1 10 100 1000 :MOR10222 N30S M54 (nM) vs 1 nM human IL18 Patent Application Publication Apr. 24, 2014 Sheet 11 of 38 US 2014/O112915 A1 Figure 5(C) 8000 6000 40 00 2 00O O h18BPa-FC 1E-3 O.O. O. f 10 100 MOR13361 (nM) vs 1nM humanil 18 1000 O hL18BPa-FC 1E-3 0.01 0.1 1 10 100 MOR13341 (nM) vs 1nM humani 18 Patent Application Publication Apr. 24, 2014 Sheet 12 of 38 US 2014/0112915 A1 Figure 6(A) 5000 A000 4000 a. 3000 S3. 3000 8e zoo 20 f 000 1000 0. h183Parc C h3BPa-FC 0.01 0. 1 10 100 1000 0.01 01 10 100 1000 MORO944 (nfM) MOR3464 (nm) 7000 5000 6000 400 5000 4000 3000 3000 2000 Z 2000 1000 1000 O h183Pa-Fc O h18BPa-Fc 0.01 0.1 1 0 100 000 0.01 0 1 10 100 000 MOR10579 (nM) MOR10222 (nW) 3000 300 2500- 3000 2000 2500 e 2000 a 1500 ab g se 1500 i 1000 fe 1000 500 500 O 0.01 0.1 1 0 100 100 10 100 hi-18BPa-Fic (nM) MoR08775 (nM) Patent Application Publication Apr. 24, 2014 Sheet 13 of 38 US 2014/O112915 A1 Figure 6(B) 20000 St 5000 e 10000 5000 O 10 100 MOR08776 (nM) hL18BPa-FC 1E-3 0.01 0,1 1 10 100 1000 MORO9464 N3OK (nM) vs 1nM cynolL18 1 OO O hL18BPa-FC 1E-3 O.01 0.1 1 0 100 1000 MOR10222 N30S M54 (nM) vs 1nM cynolt 18 Patent Application Publication Apr. 24, 2014 Sheet 14 of 38 US 2014/O112915 A1 Figure 6(C) 500 O 0.01 0. 1 10 100 MOR13361 (nM) vs 1nM cynolL18 1000 O hL18BPa-FC 1E-3 0.01 0.1 10 100 MOR13341 (nM) vs 1nM human? 18 Patent Application Publication Apr. 24, 2014 Sheet 15 of 38 US 2014/O112915 A1 Figure 7(A) 3000 3500 30 2500 so f 150 1000 1000 hi-18BPa-FC 500 hi-18BPa-FC 01 0. 1 0 100 000 0.01 O. 10 100 000 MORD9441 nM. MORO9464 (n) f 1 hi-18BPa-FC 801 0. 0 0 100 0.01 . 1 100 to i MOR10579 (nivl) MOR10222 (nM) 300 250 50 h33Pa-Fe 1 OO 183Pa-Fc 50 0. o 00 000 O.1 O 00 NORO9465 (nm) MORO9466 (nRM) Patent Application Publication Apr. 24, 2014 Sheet 16 of 38 US 2014/O112915 A1 Figure 7(B) 5000 500 AO00 12so 1000 g 3000 E 750 - 2000 2 ...i 500 hi-18BPa-FC 1000 250 hi-18BPa-FC O iss- O 0.01 0 1 0 100 000 0.01 .1 1 0 100 to MOR10497 (nM) MOR10501 (nivl) 1500 400 1250 3500 in 1000 3000 a 2500 3 750 R2000 Nowa e 500 hiL-13BPa-Fc a 1500 250 000 SOO 0. O.O. O. 1 10 00 000 0.0 O, O 100 000 MOR0502 (nM) hit -18 BP-Fo (nW) Patent Application Publication Apr. 24, 2014 Sheet 17 of 38 US 2014/O112915 A1 Figure 7(C) 500 100 – hiL18BPa-FC 0.01 0,1 1 0 100 1000 MORO9464 N3OK(nM) 0.01 0.1 10 OO 1000 MOR10222 N30S M54 (nM) Patent Application Publication Apr.