US 2016.0030594A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0030594 A1 ABRAMS et al. (43) Pub. Date: Feb. 4, 2016 (54) ANTIBODY DRUG CONJUGATES Related U.S. Application Data (71) Applicants: Tinya ABRAMS, Acton, MA (US); (60) Provisional application No. 61/793,641, filed on Mar. Steven COHEN, San Diego, CA (US); 15, 2013. Christie P. FANTON, Oakland, CA EEEE): Publication Classification Kathy MILLER, San Francisco, CA (51) Int. Cl (US); Siew Ho SCHLEYER, El Cerrito, we CA (US); Kathrin Ulrike g 7. :08: TISSOT-DAGUETTE, Planegg (DE) A647/12 (2006.01) (72) Inventors: Tinya ABRAMS, Acton, MA (US); A647/26 (2006.01) Steven COHEN, San Diego, CA (US); A 6LX3/537 (2006.01) Christie P. FANTON, Oakland, CA A6II 45/06 (2006.01) (US); Catrin FINNER, Neuried (DE); (52) U.S. Cl. Thomas HUBER, Allschwil (CH): CPC. A61 K47/48561 (2013.01); A61K 47/48384 Kathy MILLER, San Francisco, CA (2013.01); A61 K3I/537 (2013.01); A61 K (US); Siew Ho SCHLEYER, El Cerrito, 45/06 (2013.01); A61 K47/12 (2013.01); A61 K CA (US); Kathrin Ulrike 47/26 (2013.01); A61 K47/4863 (2013.01); TISSOT-DAGUETTE, Planegg (DE) A61K47/48592 (2013.01); A61K 47/48615 (2013.01); C07K 16/32 (2013.01); C07K (21) Appl. No.: 14/774,512 231 7/565 (2013.01); C07K 231 7/24 (2013.01) (86). PCT No.: PCT/US14/24597 The present invention relates to anti-cKIT antibodies, anti S371 (c)(1), body fragments, antibody drug conjugates, and their uses for (2) Date: Sep.10, 2015 the treatment of cancer. Patent Application Publication Feb. 4, 2016 Sheet 1 of 53 US 2016/0030594 A1 Patent Application Publication Feb. 4, 2016 Sheet 2 of 53 US 2016/0030594 A1 ZGIRLÍTÐIH Patent Application Publication Feb. 4, 2016 Sheet 3 of 53 US 2016/0030594 A1 FIGURE 3 Sensitivy to c-Kit ADC by indication Celine Sensitivity to 9P3 C. es f t s kis s o re s 's se 9F3-CC- Gis at Patent Application Publication Feb. 4, 2016 Sheet 4 of 53 US 2016/0030594 A1 FIGURE 4 c-Kit ADC activity on GIST TI cells ABC Activity on 3S3-fi arrig3 is $81 SE3834-3C . 83-SC-8: 38-3CC -S-8: i8-8:C-: ias a 3. 8-i-. as a SS-88 0% to 88: . 8: . i s: 8. ADC 8 Patent Application Publication Feb. 4, 2016 Sheet 5 of 53 US 2016/0030594 A1 FIGURES c-Kit ADC activity on GIST 430 cells AiC Activity on GiST-430 80% 88, 8 -His3-88CC-Ra: s was as 'K328-8-8 80% assis-R-8 R ama G3-8-8 assesse SSSCSS-CC & R axas SSS8-8-8 48% - Y, ww.--SGC8-3-8 are 28-385 2 : . 0% to 8 : 3. t i E3 Aix in: Patent Application Publication Feb. 4, 2016 Sheet 6 of 53 US 2016/0030594 A1 FIGURE 6 c-Kit ADC activity on NCI-H526 cells (Small cell lung cancer (SCLC ABC Activity on NC-H526 -- isg-3C-8 ser:S328-3-8: see E88S-8-8 seeeeeee-SSS-8s senses:03-88-3 a -3803-38C-38 atta E-8-8 as a 38-C-E8. s: 88 i. 8. : 8. ASC if Patent Application Publication Feb. 4, 2016 Sheet 7 of 53 US 2016/0030594 A1 FIGURE 7 c-Kit ADC activity On NCI-H1048 cells ABC Activity of iCi-hit 848 Cells 100% 3. ww.g3-8:CC-83) areer-es-C-8 888 w attal:S388-38C,83 eneeeeee:S388-8-8 Wr as a R38-3-88 3% C.R. 8: S. AEE is is Patent Application Publication Feb. 4, 2016 Sheet 8 of 53 US 2016/0030594 A1 FIGURE 8 c-Kit ADC activity on CMK-1 1-5 cells AC Activity on Citk-11-5 refrig3-$83-8: see 8-8-8 see: SS-S-3 r & EGE9-3C-s: 8 : 8 3. AEC inf: Patent Application Publication Feb. 4, 2016 Sheet 9 of 53 US 2016/0030594 A1 FIGURE 9 c-Kit ADC activity on Ukc-1 cells AEC Activity on 'ke 83% ------------------------------------------------------------------ Y - V ---------- wagg-ScC-88; 83% -------------------------------------------------------------------------------------------------------------------------- asses.as series,8-3-3 48% ---------------------------------------------------------------------- me a 383-88-8; 28% ---------------------------------------------------------------------------------------------------------------------N- RSR i: 8. s 38 s i AC is Patent Application Publication Feb. 4, 2016 Sheet 10 of 53 US 2016/0030594 A1 FIGURE 10 Mapping of anti-c-Kit binding to C-kit receptor Baseline Corrected Eifferences f$tti irr 33 . Regic: ; Region 2 $ : : } sss * estige position Patent Application Publication Feb. 4, 2016 Sheet 11 of 53 US 2016/0030594 A1 ??c?õægNz ILGIRLÍTÐIH Patent Application Publication Feb. 4, 2016 Sheet 12 of 53 US 2016/0030594 A1 ZIGIRLÍTÐIH Patent Application Publication Feb. 4, 2016 Sheet 13 of 53 US 2016/0030594 A1 FIGURE 13 NEG085 and 20376 mediate rapid internalization of c-Kit on cells A As S- B 20.03% co- Keye A: A Create 1208 g 100.00% are rates s -- NEG85 Origin. 80.8 . 80.33%, i. 8 was 38 5 48.8% easy;3333 2.s 43 g 2.33% .3%& : : . r. : . is 3 8 24 8 4. 8 :2 8 20 2. it is:ubaida:3: ite atej AC fearet Patent Application Publication Feb. 4, 2016 Sheet 14 of 53 US 2016/0030594 A1 Patent Application Publication Feb. 4, 2016 Sheet 15 of 53 US 2016/0030594 A1 FIGURE 15 Activity of NEG085, NEG 024, 20376 in Proliferation ASSavs. In SCF-Dependent Cell Line Mo7e Ab and ADC activity or SCF-dependent proliferation re-engis error SSS a 4 g3-8:30-88 as is 8S-i- ressesses. 35 assas: - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - for a deer 88: 83C: 33 3. - 3. 8 AOC is Patent Application Publication Feb. 4, 2016 Sheet 16 of 53 US 2016/0030594 A1 FIGURE 16 Activity of NEG085 in Proliferation ASSays in Cell Line Mo7e, treated with GM-CSF Ab and ABC effect on G-CSF dependent prošiferation of Bige ces wragg arease SS88s a tra g3-...Q-E) : was leg 383-C-3 . 8. 3.338 ... is .: 3. 8. Ah or AEC & Patent Application Publication Feb. 4, 2016 Sheet 17 of 53 US 2016/0030594 A1 FIGURE 17 NEGO85 and 20376 do not mediate in vitro ADCC RK33 WS ke a rtr 836 are 85. strespontaneous reseasis as Saisipat: 8: B.E. E. 33 . cg QBE, gii. Patent Application Publication Feb. 4, 2016 Sheet 18 of 53 US 2016/0030594 A1 FIGURE 18 cKIT antibodies do not lead to apoptosis of human primary mast cells A EC. rhi NEG885 & 38 98 a 28378 & 38 88 y ig&S > 8. i 8, & o-c-kit Ab sit Patent Application Publication Feb. 4, 2016 Sheet 19 of 53 US 2016/0030594 A1 FIGURE 19 NEG085 and 20376 do not mediate mast cell degranulation on ex vivo primary human mast cclls siast Celegranulation i-hexosarainidase activity 388 : 330303 BSEC$383 ' R{x}{{R}} s Sigg 8:38 i : & 23 F8 20,000 GC8S e ; Si3O3(K. 8. Ef & ECC: SNP (58SA Six8 S SS 83. Patent Application Publication Feb. 4, 2016 Sheet 20 of 53 US 2016/0030594 A1 FIGURE 20A Co-localization of IgG1 and mitotic arrest of NEG027-MCC-DM1 in GISTT1 xenograft model FIGURE 20B P-HistOne H3 Patent Application Publication Feb. 4, 2016 Sheet 21 of 53 US 2016/0030594 A1 IZGIRLÍTÐIH Patent Application Publication Feb. 4, 2016 Sheet 22 of 53 US 2016/0030594 A1 FIGURE 22 Mitotic arrest and apoptosis induction 8 days post single dose of cKIT ADC Single 2.5 mg/kg IV dose of NEGO27-MCC-DM1 i -N- NEGQ27-vic-Mi', R-Histore H3 s -- NEG027-MCC-N1% Ceawed Caspase 3 -er vehicle % p-Histone H3 5 war Wehicle's Ceawed Caspase 3 18 Days Post-ose Patent Application Publication Feb. 4, 2016 Sheet 23 of 53 US 2016/0030594 A1 FIGURE 23 (A) DOSe response efficacy in GIST T1 mouse Xenograft One large tumor 1 5 O O removed Buffer isotype ControllgG1-MCC-DM1 2.5 mg/kg NEG027-MCC-DM1 0.625 mg/kg NEG027-MCC-DM1 1.25 mg/kg NEGO27-MCC-DM1 2.5 mg/kg 5 O O O 1 O 2O 30 40 50 60 Days Post-Implant D O S e d B) Change in body weight over course of treatment - Eiff or isotype Controi iggi-RCC-ilt 2.3 pagik 2 -- REG02iticC-8; 2. Singkg -- NEG027. Ricc-Bi (),825 agkg -- RES027-i(-tt .25 mgkg 1 3. s: 3. gigs Post-gar Ocsec. y g Patent Application Publication Feb. 4, 2016 Sheet 24 of 53 US 2016/0030594 A1 FIGURE 24 (A) anti-DM1 ELISA after dosing in a GIST T1 xenograft model 1OOOOO -- NEG027-MCC-DM1 0.625 mg/kg -- NEG027-MCC-DM1 1.25 mg/kg -0- NEG027-MCC-DM1 2.5 mg/kg 1OO O 5 10 15 20 25 Days Post-Dose (B) anti-human IgG1 ELISA after dosing in a GIST T1 xenograft model 1OOOOO -A- NEG027-MCC-DM1 0.625 mg/kg -0- NEG027-MCC-DM1 1.25 mg/kg 1OOOO -o- NEG027-MCC-DM1 2.5 mg/kg 1OOO 100 O 5 10 15 2O 25 Days Post-Dose Patent Application Publication Feb. 4, 2016 Sheet 25 of 53 US 2016/0030594 A1 $ $${}% SZCHRI[10IH Patent Application Publication Feb. 4, 2016 Sheet 26 of 53 US 2016/0030594 A1 FIGURE 26A NEG027-MMC-DM1 dose response efficacy in GISTT1 Group 4 pooled tumors FIGURE 26B NEG027-MMC-DM1 dose response efficacy in GISTT1 Group 5 pooled tumors Patent Application Publication Feb.