US 2015 0037355A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0037355A1 Kirn et al. (43) Pub. Date: Feb. 5, 2015 (54) GENERATION OF ANTIBODIES TO TUMOR A 6LX39/235 (2006.01) ANTIGENS AND GENERATION OF TUMIOR A6135/26 (2006.01) SPECIFIC COMPLEMENT DEPENDENT GOIN33/574 (2006.01) CYTOTOXCITY BY ADMINISTRATION OF C07K 6/8 (2006.01) ONCOLYTIC VACCNAVIRUS A6II 45/06 (2006.01) CI2N 7/00 (2006.01) (75) Inventors: David Kirn, Mill Valley, CA (US); John A 6LX39/395 (2006.01) Bell, Ottawa (CA); Caroline Breitback, (52) U.S. Cl. San Francisco, CA (US); Anne Moon, CPC ............. A6IK39/285 (2013.01); A61K.39/395 San Francisco, CA (US); Tae-Ho (2013.01); A61 K39/12 (2013.01); A61 K Hwang, Busan (KR); Yu Kyoung Lee, 39/145 (2013.01); A61K 39/165 (2013.01); Busan (KR); Mi-kyung Kim, Busan A61K39/17 (2013.01); A61 K39/125 (KR) (2013.01); A61K 39/205 (2013.01); A61 K 39/245 (2013.01); A6IK 39/235 (2013.01); (73) Assignees: SILLAJEN, INC., Busan (KR): A6135/26 (2013.01); GOIN33/574 JENNEREX INC., SAN FRANCISCO, (2013.01); C07K 16/18 (2013.01); A61K 45/06 CA (US) (2013.01); C12N 7/00 (2013.01); C12N 2710/10034 (2013.01); C12N 27.10/16634 (21) Appl. No.: 13/978,113 (2013.01); C12N 27.10/24034 (2013.01); C12N 2760/18734 (2013.01); C12N 2760/18434 (22) PCT Fled: Jan. 4, 2012 (2013.01); C12N 2760/18134 (2013.01); C12N PCT NO.: PCT/US12/20173 2760/20034 (2013.01);s C12N 2760/16034 (86) O (2013.01); C12N 2770/36034 (2013.01); C12N S371 (c)(1), 2770/32034 (2013.01); C12N 2770/32334 (2), (4) Date: Oct. 24, 2014 (2013.01); C12N 2750/14034 (2013.01); A61 K 2039/507 (2013.01) Related U.S. Application Data USPC .................. 424/174.1; 424/204.1; 424/209.1; (60) Provisional application No. 61/429,622, filed on Jan. 424/212.1; 424/214.1; 424/216.1; 424/218.1; 4, 2011. 424/224.1; 424/230.1; 424/231.1; 424/232.1; s 424/233.1; 424/93.71; 435/375:506/9; Publication Classification 5307389.7 (51) Int. C. (57) ABSTRACT A 6LX39/285 (2006.01) The present invention relates to methods and compositions A6 IK 39/2 (2006.01) for use in inducing tumor-specific antibody mediated A6 IK39/45 (2006.01) complement-dependent cytotoxic response in an animal hav A6 IK39/65 (2006.01) ing a tumor comprising administering to said animal a com A 6LX39/7 (2006.01) position comprising a replication competent oncolytic virus A 6LX39/25 (2006.01) wherein administration of the composition induces in the A 6LX39/205 (2006.01) animal production of antibodies that mediate a CDC response A6 IK 39/245 (2006.01) specific to said tumor. Patent Application Publication Feb. 5, 2015 Sheet 1 of 26 US 2015/0037355A1 FIGURE 1A m Sessi 'sy X-53s reise $3888 assi: Sssssss for SS is scis WX $3e3i's assist Sessis iss's X-S8s res:Še XX 88s: six Š S& Rai SC Patent Application Publication Feb. 5, 2015 Sheet 2 of 26 US 2015/0037355A1 FIGURE 1B s S 3. 3S 3. says post irection Patent Application Publication Feb. 5, 2015 Sheet 3 of 26 US 2015/0037355A1 FIGURE 1C 2 won S is ciersy mcame is is sessity is . .8. 8 is . 8 Seri case is Patent Application Publication Feb. 5, 2015 Sheet 4 of 26 US 2015/0037355A1 FIGURE 1D D VX2 lysates PBMC lysates 2 Nave at Sei 2. 3W post JX-594 into VX2 bearing rabbit Patent Application Publication Feb. 5, 2015 Sheet 5 of 26 US 2015/0037355A1 FIGURE 2A - FIGURE 2B s S & s ex & 's s 3. Patent Application Publication Feb. 5, 2015 Sheet 6 of 26 US 2015/0037355A1 FIGURE 3A swsS. S is 888& $$$$$. : - S : is sexists: $88: is $88ws $88.8 xxi.88. Patent Application Publication Feb. 5, 2015 Sheet 7 of 26 US 2015/0037355A1 FIGURE 3B 3 is $8'3: x set 38: S: s s **** 2º:32.§ c.k& Patent Application Publication Feb. 5, 2015 Sheet 8 of 26 US 2015/0037355A1 FIGURE 3C "b" serum "c" serum d" serum Patent Application Publication Feb. 5, 2015 Sheet 9 of 26 US 2015/0037355A1 FIGURE 3D 2O O 8 O 8 O 4. O 2 O O 5 O 3O Time of Heat Treatment (Min at 56 °C) Patent Application Publication Feb. 5, 2015 Sheet 10 of 26 US 2015/0037355A1 FIGURE 4A #301-RCC pt. SSSSSSSSS Patent Application Publication Feb. 5, 2015 Sheet 11 of 26 US 2015/0037355A1 FIGURE 4B #304 --melanoma pt. m g s: 5 s E wy & { ge is is is 120 y SS. m19 in &e (y s 100 S Š 80 60 40 s Patent Application Publication Feb. 5, 2015 Sheet 12 of 26 US 2015/0037355A1 FIGURE 4C if 2. HCC pt, S &SS Patent Application Publication Feb. 5, 2015 Sheet 13 of 26 US 2015/0037355A1 FIGURE 4D Patent Application Publication Feb. 5, 2015 Sheet 14 of 26 US 2015/0037355A1 FIGURE 5A i8.8s2 i S. s A2780 cell viability (CDC activity) is of JX-39s injected sets baseine serin Patent Application Publication Feb. 5, 2015 Sheet 15 of 26 US 2015/0037355A1 FIGURE 5B 100 - Heps2HeroG2 B SS JNU449 8. N “res s is SNU475 s: SNU739. x i ( 20 S 3 SS is is 8. 8.S 7 Post X-584 injection Days Patent Application Publication Feb. 5, 2015 Sheet 16 of 26 US 2015/0037355A1 FIGURE 5C C 5 ( 5 O S 3 35 it 5 is 55 S 65 Post JX-594 injection Days) Patent Application Publication Feb. 5, 2015 Sheet 17 of 26 US 2015/0037355A1 FIGURE 5D SNU475 x N. Y. 8 8, 5 O 5 25 3. 35 is 45. 5. 55 SO 85 7. Post JX-584 injection (Days Patent Application Publication Feb. 5, 2015 Sheet 18 of 26 US 2015/0037355A1 FIGURE 5E - was 2-&s^ ^ y SNU449 m &^. ai, . 25 3. 35 is is 55 3 SS Post JX-594 injection (Days) Patent Application Publication Feb. 5, 2015 Sheet 19 of 26 US 2015/0037355A1 FIGURE 6 S Patent Application Publication Feb. 5, 2015 Sheet 20 of 26 US 2015/0037355A1 FIGURE 7A -O JX-594 naive Serum -O- JX-594 injected serum O.O O.2 O.4 Sunitinib dose (ug/mL) Patent Application Publication Feb. 5, 2015 Sheet 21 of 26 US 2015/0037355A1 FIGURE 7B 120 6 O 40 -O- JX-594 injected Serum 2O al-O JX-594 naive Serum O.O O.5 1.O 15 2.O Sorafenib dose (ug/mL) Patent Application Publication Feb. 5, 2015 Sheet 22 of 26 US 2015/0037355A1 FIGURE 7C --a && S&S$388 & S&isiy Patent Application Publication Feb. 5, 2015 Sheet 23 of 26 US 2015/0037355A1 FIGURE 8 CDC profile after JX-594 ..a a. treatment in CC patients. Red colored serum (six patients) were pooled for SEREX screening is is R$ 3: 3& S. 38 is S as S Post X-SS: isjeciati jays wwy sts as st cs: ines - artial caNA phage librar E.gif ytic infection, &age; it assifer to serixate & Essay scsesitg with pixie Sier8 &ciexi settish : sesses&e exi ser:Y is it. ge3: & 33six & & {x&iss aftes 2 xis: y scissisi'g. is $38ises 38&ia &Sig's Cississ so sistig sig &38 S33& &ise $38,383 & 888 pityay streeting, Ess&ix spie is post 38 Sgiates x 3 sti&stigs Eiky 533 injectics SSSSSSSSSSSSSS&S Patent Application Publication Feb. 5, 2015 Sheet 24 of 26 US 2015/0037355A1 FIGURE 9 150 : as PBS serum(n=2) hGM-CSF594 serum(n=2) OO -- uv inactivated hCGM-CSF594 serum(n=2) eye mM-CSF594 serum(n=2) -8- JX-963 serum(n=2) ox- Reovirus serum(n=2) O s s s 8 8. S ° s” N Serum (%) FIGURE 10 150 \&s JX594 Serum -8. JX594mGM-CSF Serum -- Control: pre treatment serum’ m$x WR Serum 50 wax VSV Serum r:8tra VSV GM-CSF Serum Serum (%) Patent Application Publication Feb. 5, 2015 Sheet 25 of 26 US 2015/0037355A1 FIGURE 11 sys HSV serum(n=2) -- VSV GM-CSF serum (n=2) Serum (%) FIGURE 12 100 leuuougoK?IIqe?A% ULIITLIÐS &&&&&&&&& virus types Patent Application Publication Feb. 5, 2015 Sheet 26 of 26 US 2015/0037355A1 FIGURE 13 's s US 2015/0037355A1 Feb. 5, 2015 GENERATION OF ANTIBODIES TO TUMIOR reproducible delivery to distant tumors were limited, how ANTIGENS AND GENERATION OF TUMIOR ever. Systemic anti-cancer potency and blood-borne delivery SPECIFIC COMPLEMENT DEPENDENT to metastatic tumors therefore had to be improved. CYTOTOXCITY BY ADMINISTRATION OF 0006 Given both the potential and the limitations with ONCOLYTIC VACCNAVIRUS each of these three individual virus-based approaches, we asked whether it was possible to combine and optimize the RELATED APPLICATIONS best attributes of each into a single therapeutic agent. Tar 0001. The present application is the 35 USC 371 U.S. geted and armed oncolytic poxviruses have the potential to do national stage of international application no. PCT/US2012/ so. JX-594 is a 3'-generation oncolytic poxvirus therapeutic 020173, filed on Jan. 4, 2012 and claims the benefit of priority designed to have three complementary MOA including: 1) of U.S. Provisional Application No. 61/429,622, which was direct replication-mediated oncolysis, and 2) active cancer vaccination. JX-594 is a Wyeth vaccinia virus vaccine-de filed on Jan.