US 2015 0337042A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0337042 A1 Reilly et al. (43) Pub. Date: Nov. 26, 2015 (54) ANTI-EGFRANTIBODIES AND ANTIBODY (22) Filed: Mar. 20, 2015 DRUG CONUGATES Related U.S. Application Data (71) Applicant: ABBVIE INC., North Chicago, IL (US) (60) Provisional application No. 61/968,819, filed on Mar. 21, 2014. (72) Inventors: Edward B. Reilly, Libertyville, IL (US); Publication Classification Andrew C. Phillips, Libertyville, IL (US); Lorenzo Benatuil, Northborough, (51) Int. Cl. MA (US); Fritz G. Buchanan, Antioch, C07K 6/28 (2006.01) IL (US); Jonathan A. Meulbroek, Lake A647/48 (2006.01) Bluff, IL (US); Chung-Ming Hsieh, (52) U.S. Cl. Newton, MA (US); Jennifer Perez, CPC ....... C07K 16/2863 (2013.01); A61K 47/48561 (2013.01); C07K 2317/21 (2013.01); C07K Granville, NY (US) 231 7/565 (2013.01) (57) ABSTRACT (73) Assignee: ABBVIE INC., North Chicago, IL (US) The invention relates to anti-epidermal growth factor (EGFR) antibodies and antibody drug conjugates (ADCs), including compositions and methods of using said antibodies and (21) Appl. No.: 14/664,453 ADCs. Patent Application Publication Nov. 26, 2015 Sheet 1 of 24 US 2015/0337042 A1 8HODZHOD|HCJD |?un6|+ Patent Application Publication Nov. 26, 2015 Sheet 4 of 24 US 2015/0337042 A1 FACS Binding to Tumor Cells 1 OOOO 7500 5OOO 25OO O.O1 1 100 1 OOOO Patent Application Publication Nov. 26, 2015 Sheet 5 of 24 US 2015/0337042 A1 009/ ueeu Oed Patent Application Publication Nov. 26, 2015 Sheet 6 of 24 US 2015/0337042 A1 EGFR (1-525) Binding kinetics (closed circles) = Ab1 and Ab2 106 (open circles) = variants Figure 6 Patent Application Publication Nov. 26, 2015 Sheet 7 of 24 US 2015/0337042 A1 SCC-15 (Ab1 Sensitive) o mAb - <C up na > a n on U a (1333 nM) - O - O - O - O - O - O - O O - O - O - O cC. C C C C C C U C C CC pYEGFR tOt EGFR Actin Š& Figure 7A H292 (Ab1 Resistant) i Figure 7B Patent Application Publication Nov. 26, 2015 Sheet 8 of 24 US 2015/0337042 A1 EEEEEEEEEEEEEEEEEEEEEEE ZAZZAZAAAAAAAAAZZAZAZZZZYZZAAA As a YYYYYYYYYYYYYYYYYYYYYYYYYY IEEEEEEEEEEEEEE ZaaZaaaaaZaA Net N. cCW sists, ZAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA NYa YaYa YaYa YaYa Yasa YaYa Ya OCW E. sists ZZZaaZZaaaaaaaaaaaaaZZ Stan-Natasanta ESSEE NCW IEEEEEEEEEEEEEEEEEEEEEEEEEE HZAZAAAAWAAAAAAAWAAAAAAAZ H See WCW IEEEEEEEEEEEEEEEE Zazaaaaaaaaaaaaaaaaaaaaaaaaaaaa. NYYYYYYYYYYY ESE ICW AAAAAZZZZZZYZZAZAZA X C W YYYYYYYY. al ZY Ess: ZaaZ-Zaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa. SSSSSSSXSYS s s KERIEEEEEEEEEEEEEEEEEEEEEEE RAAAZZZZAAAAAZZZZAAAA HYYYYY Y.Y. is: OCW HEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEEE 24 CW as4. Ya Ya X Y. Ya Ya N. Y. W. S. Ya Ya NYS X Ya Ya Ya Ya Ya Y. Ya. ESSESSESSESSESSESSESS CW EEEEEEEEEEEE AAZZY CCW E. s YaaaaaaZaaZZZZZYaaaaaaZaaZZ a Y XYY OCW w HEBREEEEEEEEEEEEEE ZAZZZZZAAAAZZZZZZZZAAAAZZZ SS reeRS NetNetNRS RS RS OCW EHEREEEEEEEEEEEEEEEE EAZAZAZZAZ HSN.Y., a Ya Ya YaYa YYYYaYa YaYa ESESSESSESSEE 9CW WCW IEEEEEEEEEEEEEEEEEE s AZWZZZZZZZZZZAAAAZZ Z s a seeae LCW n CO S O s O O O Patent Application Publication Nov. 26, 2015 Sheet 9 of 24 US 2015/0337042 A1 125 1 OO SC 75 Ab1 50 A AbP & IC5o Y 40 nM 25 V Ab2 O O.1 1 1 O 1 OO 1 OOO 1 OOOO Ab (nM) Figure 8B Patent Application Publication Nov. 26, 2015 Sheet 10 of 24 US 2015/0337042 A1 NHEK 5OOO 45OO 4OOO 3500 a Ab 1 3OOO O AbA 2500 A. (g 2000 O AbM 1500 AbP 1 OOO Ab2 500 A Control 1 O O.O1 O.1 1 10 100 1 OOO 10000 Ab (nM) Figure 9 Patent Application Publication Nov. 26, 2015 Sheet 11 of 24 US 2015/0337042 A1 NC-H292 Human NSCLC Carcinoma Xenograft 2OOO 1 5 O O 1 OOO 500 10. 20 3O Day post tumor cell injection Figure 10 Patent Application Publication Nov. 26, 2015 Sheet 12 of 24 US 2015/0337042 A1 Patent Application Publication Nov. 26, 2015 Sheet 13 of 24 US 2015/0337042 A1 AAGA 699 09| GZ| Patent Application Publication Nov. 26, 2015 Sheet 14 of 24 US 2015/0337042 A1 A/COL Z-ZL?un61 08Z@96ed/'86 ZHV/C] Patent Application Publication Nov. 26, 2015 Sheet 17 of 24 US 2015/0337042 A1 NCI-H1703 (NSCLC) esa 27OO -O-Ab1---- $2. (10 2m Smka) 2400 - O - Ab1-mcMMAF (1 mkd) 2100 -H AbA-vcMMAE (1 mkd) o E O D o E E C ? CU > 28 35 42 49 56 63 70 77 84 91 98 1051 12 Day post tumor cell inoculation Figure 14A EBC1 (NSCLC) 28OO --O-- Control 2 (10 mkd) -(- Ab 1 a - 2400 -O-Ab3 mcMMAF 3mkd --- Ab1-mcMMAF E 2000 -H AbA mcMMAF g 5 1600 O D 5 1200 8 800 C ?t 3 400 217 3-O-O-O---------------------------1------------- 7 14 21 28 35 42 49 56 63 70 77 81 Day post tumor cell inoculation Figure 14B Patent Application Publication Nov. 26, 2015 Sheet 18 of 24 US 2015/0337042 A1 -o-Control 2 (6) NC-H292 "O" Control 2 voMMAE (3) 2400 -- Ab1 voMMAE (3) -H Control 2 mcMMAF (3) -o- Ab1 voMMAEp (3) -N- Ab1-mcMMAF (3) 2100 -- AbAvcMMAE (3) --O-- Ab1-mcMMAFp (3) -0-- AbAvcMMAEp (3) S 1800 1500 1200 900 3OO y 3 10 17 24 31 38 45 52 59 66 73 80 87 94101108 Days post cellinoculation Figure 15A -o-Control 2 (6) NC-H292 2400 rom Control 2 voMMAE (6) -H Control 2 mcMMAF (6) -- Ab1 voMMAE (6) -N- Ab1-mcMMAF (6) 2100 -o- Ab1 vcMMAEp (6) --O- Ab1-mcMMAFp (6) -- AbAvcMMAE (6) 1800 --- AbAvcMMAEp (6) 1500 : 12OO 900 300 3 10 17 24 31 38 45 52 59 66 73 80 87 94 101108 Days post cellinoculation Figure 15B Patent Application Publication Nov. 26, 2015 Sheet 19 of 24 US 2015/0337042 A1 £HODZHOD|HCJD Patent Application Publication Nov. 26, 2015 Sheet 20 of 24 US 2015/0337042 A1 6 1 -Ligand binding Deleted in domain (L1) EGFRV domain (CRS Receptor dimerization -Ligand binding domain III (L.2) -Receptor dimerization domain IV (CR2) transmembrane juxtamembrane A: Ab1 epitope OB: region recognized Ab2 tyrosine kinase regulatory region 1186 Figure 17 Patent Application Publication Nov. 26, 2015 Sheet 21 of 24 US 2015/0337042 A1 NC-H292 (NSCLC) 2400 mon. Control 2 (6) -H Control 2 mcMMAF (6) -o-, Control 2 WCMMAE (6) - HAb1-mcMMAF (3) 209-o-AbA-vcMMAEp (3) -2. Ab1-mcMMAF (6) -o- AbA-vcMMAEp (6) 1 8 O O 1500 12OO 900 300 3 10 17 24 31 38 45 52 59 66 73 80 87 94 101108 Days post cell inoculation Figure 18 Patent Application Publication Nov. 26, 2015 Sheet 22 of 24 US 2015/0337042 A1 U87MGde2-7 (GBM) 28OO -O-Control 2 (1) -0. AbA-MMAEp (1) 2400 --O-- Ab1-mcMMAF (1) - O - AbA-mcMMAF (1) 2OOO 1600 12OO 8OO 400 7 14 21 28 35 Days post cell inoculation Figure 19 Patent Application Publication Nov. 26, 2015 Sheet 23 of 24 US 2015/0337042 A1 -o- IgG Control 3 --O--- Ab 1 - A - AbA O 24 48 72 96 12O 144 168 FigureTime (hrs) 20A -o-lgG Control ----- Ab1 0.81-A - AbA 0 24 48 72 96 12O 144 168 FigureTime (hrs) 20B Patent Application Publication Nov. 26, 2015 Sheet 24 of 24 US 2015/0337042 A1 *HDO HH :·C••C·SOO (Duu US 2015/0337042 A1 Nov. 26, 2015 ANT-EGFRANTIBODIES AND ANTIBODY EGFR correlates or is associated with poor prognosis of the DRUG CONUGATES patients. (Herbst and Shin, Cancer 94:1593-1611 (2002); and Modjtahedi et al., Br. J. Cancer 73:228-235 (1996)). EGFR is RELATED APPLICATIONS also expressed in the cells of normal tissues, particularly the epithelial tissues of the skin, liver, and gastrointestinal tract, 0001. This application claims the benefit of priority to although at generally lower levels than in malignant cells U.S. Provisional Application No. 61/968,819, filed on Mar. (Herbst and Shin, Cancer 94:1593-1611 (2002)). 21, 2014. The contents of the aforementioned priority appli 0006. A significant proportion of tumors containing cation are incorporated by reference herein in their entirety. amplifications of the EGFR gene (i.e., multiple copies of the EGFR gene) also co-express a truncated version of the recep SEQUENCE LISTING tor (Wikstrandet al. (1998).J. Neurovirol. 4, 148-158) known 0002 The instant application contains a Sequence Listing as de2-7 EGFR, AEGFR, EGFRVIII, or A2-7 (terms used which has been submitted electronically in ASCII format and interchangeably herein) (Olapade-Olaopa et al. (2000) Br. J. is hereby incorporated by reference in its entirety. Said ASCII Cancer. 82, 186-94). The rearrangement seen in the de2-7 copy, created on Mar. 12, 2015, is named 117813-06202 SL. EGFR results in an in-frame mature mRNA lacking 801 txt and is 110,863 bytes in size. nucleotides spanning exons 2-7 (Wong et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89,2965-9: Yamazaki et al. (1990) BACKGROUND OF THE INVENTION Jpn. J. Cancer Res. 81, 773-9: Yamazaki et al. (1988) Mol. 0003. The human epidermal growth factor receptor (also Cell. Biol. 8, 1816-20; and Sugawa et al. (1990) Proc. Natl. known as HER-1 or Erb-B1, and referred to herein as Acad. Sci. U.S.A. 87, 8602-6). The corresponding EGFR “EGFR) is a 170 kDa transmembrane receptor encoded by protein has a 267 amino acid deletion comprising residues the c-erbB protooncogene, and exhibits intrinsic tyrosine 6-273 of the extracellular domain and a novel glycine residue kinase activity (Modjtahedi et al., Br. J.