US 20150374790A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0374790 A1 Liu et al. (43) Pub. Date: Dec. 31, 2015 (54) IL-15-BASED MOLECULES AND METHODS Publication Classification OF USE THEREOF (51) Int. Cl. (71) Applicant: ALTOR BIOSCIENCE A638/20 (2006.01) CORPORATION, Miramar, FL (US) A 6LX39/395 (2006.01) A647/48 (2006.01) (72) Inventors: Bai Liu, Cooper City, FL (US); Peter (52) U.S. Cl. Rhode, Miami, FL (US), Wenxin Xu, CPC ....... A61K 38/2086 (2013.01); A61K 47/48.276 Pembroke Pines, FL (US); Hing C. (2013.01); A61K 47/48369 (2013.01); A61 K Wong, Weston, FL (US) 39/39558 (2013.01) (21) Appl. No.: 14/755,989 (22) Filed: Jun. 30, 2015 (57) ABSTRACT Related U.S. Application Data The invention features combination therapies using an IL-15 (60) Provisional application No. 62/018,899, filed on Jun. based Superagonist complex and an antibody to effectively 30, 2014. treat Subjects with cancer and infectious diseases. Patent Application Publication Dec. 31, 2015 Sheet 1 of 31 US 2015/0374790 A1 S. V d U CU 2 O A. r's O O.O1 O.1 1 10 ALT-803 (nM) Fig. 1A 50 S 40 V d 30 U g 20 3 10 O Y O O.O1 O.1 1 10 ALT-803 (nM) Fig. 1B 75 --C) - Ctrl CD4 g 60 --O- - Ctrl CD8 45 ---A--- Ctrl NK V 30 age 8O3 CD4 V 15 -- 803 CD8 U -- 8O3 NK O 3 5 7 10 CultureTime (days) Fig. 1C Patent Application Publication Dec. 31, 2015 Sheet 2 of 31 US 2015/0374790 A1 60--------------------------------------------- --K) - Ctrl CD4 Š --O-- Ctrl CD8 ---A--- Ctrl NK 3 are 803 CD4 g -- 803 CD8 S. arra 8O3 NK O 3 5 7 10 CultureTime (days) Fig. 1D 8 -o-Medium X------- S. montheme ALT-803 w do S.V U CultureTime (days) Fig.1E 2.5------------------------------------------------------------------- O 2.0----------------eas ----------------------------1----------- 1.5-------1----------------NS--(Y------------------ 3 U 1.O res. R orc) or Medium s 305 -A-ALT-803 O.O O 3 5 7 10 CultureTime (days) Fig.1F Patent Application Publication Dec. 31, 2015 Sheet 3 of 31 US 2015/0374790 A1 2.4 {d 2.0 s1. 16 W V O K) 1.2 :A V 30.8 * V V A t 04 &E C> E a O.O i? 3 n 89 n 89 CU F CU F > CC > C > CC PMBCS CD8 T cells CD4 T cells Donor 1 ADonor2 VDonor3 (>DonorA. ODOnors XDOnore Donor7 Mean SE Fig. 2A 0.2O 0.15 {d - 5. G W Se V X, 0.10 {X V S C> 0.05 : I * A A. O O o O o O.OO r ry 3 E n 3 E n 3 9CU F CU F > CC > CC > CC NK cells CD19 B cells T-reg cells DDOnOr1 ADOnOr2 VDOnors god DOnOr4 ODonors XDonoré Donor7 - Mean SE Fig.2B Patent Application Publication Dec. 31, 2015 Sheet 4 of 31 US 2015/0374790 A1 360---------------------------------------- s 100 240 mm . s 120 Q 50 S120 Y - - - - - - - - - - - - - - - - G 25 60--------------1---------- a-------- O O.O1 O.1 1 10 O O.O1 O.1 1 10 Donor-A ALT-803 (nM) Donor-B ALT-8O3 (nM) Fig. 3A Fig. 3B O 0.01 0.1 1 10 O 0.01 0.1 1 10 Donor-A ALT-803 (nM) Donor-B ALT803 (nM) Fig. 3C Fig. 3D Patent Application Publication Dec. 31, 2015 Sheet 5 of 31 US 2015/0374790 A1 O O.O1 O.1 1 10 O O.O1 O.1 1 10 ALT-803 (nM) ALT-803 (nM) Fig. 4A Fig. 4B NK cells 4000------------------------------------------ O O.O1 O.1 1 10 0 001 ol 1 10 ALT-803 (nM) ALT-803 (nM) Fig. 4C Fig. 4D Patent Application Publication Dec. 31, 2015 Sheet 7 of 31 US 2015/0374790 A1 80 Patent Application Publication Dec. 31, 2015 Sheet 8 of 31 US 2015/0374790 A1 --ALT-803 --KX-Medium 1 5 - - - - - - - - - - 10-------------- ...-- rap amm-e------------0 O:1 2:1 5:1 10:1 0:1 2:1 5:1 10:1 Fig.6AE.T Ratio Fig.6BE.T Ratio 90 s 75 is 60 C Y asO 45 ' Y -- ALT-803 30|r / Y- --K). Medium 15 - - - - - M --"e --------------------------- O O:1 2:1 5:1 10:1 E.T Ratio (with Rituximab) ALT-803 Medium Fig.6C Fig.6D Patent Application Publication Dec. 31, 2015 Sheet 9 of 31 US 2015/0374790 A1 7 5 O O O.O1 O.1 1 10 ALT-803 (nM) Fig. 7A O O.O1 O.1 1 10 ALT-803 (nM) Fig. 7B Patent Application Publication Dec. 31, 2015 Sheet 10 of 31 US 2015/0374790 A1 80 ... ' -----------------"r 80 - .is a wa was s560 "r^ ------Rituximabarea Control sg60 AA --Control...Rituximab Á40------------------------------------ 840 O O 9208 on J- G 20 O O O.O. O.O1 O. 1 1 1 O O.O. O.O1 O.1 1 1 O ALT-803 (nM) ALT-803 (nM) Fig.8A Fig.8B at-a-w/o mAb --w/o mAb 75------Rituximab -------------- s 75-------Rituximab --------------- Š 60 I-A-HOAT- - -y N 60-------------------------------- it 45- ------------------- f 45 30.- ... "NK ... 30 Non-NN 15 3 15 O O 0.01 O.1 O O.O1 0.1 ALT-803 (nM) ALT-803 (nM) Fig. 8C Fig.8D Patent Application Publication Dec. 31, 2015 Sheet 11 of 31 US 2015/0374790 A1 75------------------------------------------ 2. - - - - In vivo & 60--------------------, -, --------------------- 1. OPBS 45------------------3- ------------------- ZAlt-803 Rituximab 30|r1,r Rituximab + ALT803 15 - - - - - - - - - - - - - - - - O 2N. N: Medium Rituximab ALT-803 Rituximab + ALT8O3 In vitro treatment Fig.9A 60 -o-24D2 50- --0--ALT-803 -o-24D2 - ALT803 & 40 g 30 S ds 20 u, 10 *:p<0.05; *:pz0.001 O 24D2 (nM) 0 0001 0.01 0.1 1 10 ALT-803 (nM) O 0.0001 0.001 0.01 0.1 1 Fig.9B Patent Application Publication Dec. 31, 2015 Sheet 12 of 31 US 2015/0374790 A1 4.O 3 O 2 O 1 O O PBS ALT-803 Rituximab Rituximab + ALT803 Treatment Fig. 10 3 O 2 O 1 O O PBS Rituximab O.2 0.1 O.O5 O.O2 N-- Rituximab (10 mg/kg) and ALT-803 Fig.11 100 g 80 PBS as E.a. ALT-803 40------- Rituximab ALT-803+ & 2 Rituximab yassasssssssssssssssssssssssssss Ó 10 20 30 40 50 Study Day Fig. 12 Patent Application Publication Dec. 31, 2015 Sheet 13 of 31 US 2015/0374790 A1 e - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Survival Curve (CT26) : ALT-803 (4 ug/mouse) - : CTLA4/PD-1 (0.2 mg/mouse) 119.hiffES5 n= 4-5 (ug/mouse,(5SEESEE? mouse, totaltotal 10ludoses). doses) 199 YYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYY -s-- PBS 8O 'in -- "v" ALT-803 a - O - ALT-803 PD-L1 5 O -Air ALT-8O3 CTLA4 aba ALT-8O3 CTLA4/PD-L1 -K-r-15 CTLA4/PD-L1 10 O O. O. O. O. O. O. O. O. O. O. O. O. O. O. O. O t- N Yn V Lr VO N OO OY O - N ryn V L? Study Days Fig. 13A VO N U survival curve (CT26) : ALT-803it-------------------- (4 ug/mouse) st 2 n 6 : a CTLA4/PD-1 (0.1 mg/mouse): ; SS rIL-15 inj:SD1-SD5/SD8-SD12: 110 (5ug/mouse, total 10 doses) ; 1 OO s as asso . - - - - - - - - - - - - - - - - - - - - - - ? is 90 --- PBS s 80 ro--- C IL15 53 | aga AT-803 E 38 -A-ALT-8O3--OCTLA4 3 - O - ALT-803+OCTLA4/PD-L1 & 29 a -v-rlL-15+OCTLA4/PD-L1 O -- O 1 O 20 30 40 50 60 7O 80 Study Day L15- ALT-803+ AT-803+ P value IL15 OCTLA4/oPDL1 AT-803 OCTLA4 OCTLA4/PD-L1 PBS 001.97 0.0012 IL15 0.0172 0.0015 IL15+0CTLA4/oPDL1 0.0089 ALT803 0.0005 ALT-803-0CTLA4 04564 Fig. 13B Patent Application Publication Dec. 31, 2015 Sheet 14 of 31 US 2015/0374790 A1 r ------------------------ ryn Survival Curve (5T33P) : ALT-803riL-15 inj(5ug/mouse (1 lug/mouse) or n y n=5 : oCTLA4/PD-L1 (0.2 mg/mouse); ---------iss----------------- 18 - -3- OCTLA4 g 39 - O - OPD-L1 is 39 ahae IL-15 60 --Va-I-15 - OCTLA4 28 ----L-15 - OPD-L1 S 30 ----- ALT-803 (0.05 mg/kg) 20 -A-ALT-803 - OCTLA4 1g &l- other ALT-803 - OPD-L1 O 10 20 30 40 50 60 Study Days Fig. 14A Survival Curve (5T33P) n n=5 ra O v - "y is --- PBS 90: t . ALT-803FP51,35g) (0.05 mg/kg) Sg - - - - - - ALT-803 (0.05 mg/kg) 40 P=0.21 i. - - - - - eles as seases - - ALT-803 (0.2 mg/kg) 9 30 --> --KX-OPD-L1 (25ug) {- A. 28 P=00021 --A-oPD-L1 (100 ug) O S. O 10 20 30 40 50 60 70 80 90 Study Day Fig.14B Patent Application Publication Dec. 31, 2015 Sheet 15 of 31 US 2015/0374790 A1 Ld-92LDTeqnL % peZeuoN % peZeuoN Ld-94LDLOOT?qnL gGI‘61 C Ln C C Ln C O O O O % peZeuoN % peZeuoN Ld-9ZLDLOOTeqnL C Ln C Ln O O % peZeuoN % peZeuoN Patent Application Publication Dec. 31, 2015 Sheet 16 of 31 US 2015/0374790 A1 : PBS E. PBS O 0.1 mg/kg ALT-803 D 0.1 mg/kg ALT-803 3 O 21 mg/kg ALT-803 21 mg/kg ALT-803 e ... als 2 O O 2. : * Day 26 Day 36 Day 26 Day 36 Fig.16A Fig.16B PBS : PBS O 0.1 mg/kg ALT-803 D 0.1 mg/kg ALT-803 60 (21 mg/kg ALT-803 340 21 mg/kgAT-803 350 s r 2S 30 S240 s 2S S 530 320 20 CU 3 5 10 10 2 s % O O 2.