THETWO TONDITIONUS 20170306038A1 TO MARTIN ( 19) United States (12 ) Patent Application Publication (10 ) Pub. No. : US 2017/ 0306038 A1 BROGDON et al. (43 ) Pub . Date : Oct. 26 , 2017 ( 54 ) COMPOSITIONS AND METHODS OF USE Publication Classification FOR AUGMENTED IMMUNE RESPONSE (51 ) Int. Cl. AND CANCER THERAPY CO7K 16 / 28 ( 2006 .01 ) (71 ) Applicants : Jennifer BROGDON , Sudbury , MA CO7K 16 / 28 ( 2006 . 01) (US ) ; Daniela CIPOLLETTA , A61K 45 / 06 (2006 . 01 ) Arlington , MA (US ) ; Glenn A61K 39 /00 ( 2006 .01 ) DRANOFF , Lexington , MA ( US ) ; A61K 39 /00 (2006 .01 ) Deborah A . KNEE . Del Mar. CA (US ): (52 ) U . S . Cl. CPC . .. .. CO7K 16 / 2878 ( 2013. 01 ) ; C07K 16 /2818 Fei WANG , San Diego , CA (US ) ( 2013 .01 ) ; A61K 45 / 06 ( 2013 .01 ) ; CO7K ( 72 ) Inventors : Jennifer BROGDON , Sudbury , MA 2317 / 74 ( 2013 . 01 ) ; C07K 2317 / 732 ( 2013 .01 ) ; (US ) ; Daniela CIPOLLETTA , CO7K 2317 / 567 ( 2013 . 01 ) ; COZK 2317 / 56 Arlington , MA (US ) ; Glenn ( 2013 .01 ) ; CO7K 2317 /51 ( 2013 .01 ) ; CO7K DRANOFF , Lexington , MA (US ) ; 2317 / 24 (2013 .01 ) ; A61K 2039 /507 (2013 . 01 ) ; Deborah A . KNEE , Del Mar, CA (US ) ; A61K 2039 /505 ( 2013 . 01 ) ; COZK 2317 / 75 ( 2013 . 01 ) ; CO7K 2317/ 92 ( 2013 .01 ) ; CO7K Fei WANG , San Diego , CA (US ) 2317/ 515 (2013 . 01) ( 21 ) Appl. No. : 15 /517 ,872 (57 ) ABSTRACT (22 ) PCT Filed : Oct . 8 , 2015 The present invention provides antibody compositions , (86 ) PCT No. : PCT/ US2015 / 054770 including , e . g ., antibodies, engineered antibodies and anti body fragments that bind to a tumor necrosis factor receptor $ 371 ( c ) ( 1 ), superfamily member ( i. e . , 18 ) , and compositions comprising ( 2 ) Date : Apr. 7 , 2017 one or more additional therapeutic agents. Provided com positions are useful in enhancing CD4 + and CD8 + T cell Related U . S . Application Data responses, and in the treatment, amelioration and prevention (60 ) Provisional application No. 62/ 061, 644 , filed on Oct . of diseases that can be counteracted with an augmented 8 , 2014 , provisional application No . 62 / 198 ,673 , filed immune response, e . g . , cancers. Also provided are methods on Jul. 29, 2015 , provisional application No . 62 / 220 , of use of combinations that find use in treatment or preven 764 , filed on Sep . 18 , 2015 . tion of cancerous or infectious conditions and disorders. Patent Application Publication Oct. 26 , 2017 Sheet 1 of 14 US 2017 /0306038 A1 120 120 120 100100 -100 _100I Bod 60 FIG.1A ? or L607 A40 u - P 0+ 0+ 0.1 04 -0.17 0.2 0.1 -0.14 -0.1 27.0 -0.2 2.0- RPTGGPCCGPGRLLLGTGTDARCCRVHTTRCCRDYPGEECCSEWDCMCVCPEFHCGDPCCTTCRHHPCPPGQGVQSGGKFSFGFQCDCASGTFSGGHEGHCKPWTDCTCFGFL 0.05 059.0- -0.15 Patent Application Publication Oct. 26 , 2017 Sheet 2 of 14 US 2017 /0306038 A1 CRD1 CRD2 CRD3 TM hGITR ecd hGITR CRD1 - tail - - hGITR CRD2 - tail - - - - - hGITR CRD - tail - - - - - - - hGITR ecd - tail - FIG . 1B - OD650nm ????????????????? hGITR ecd CRD1 - tail CRD2 - tail CRD3 - tail tail MAB4 MAB5 Control MAB FIG . 1C Patent Application Publication Oct. 26 , 2017 Sheet 3 of 14 US 2017 /0306038 A1 PBS hGITRecdmFc R90A R59A R56A R51A R48A R38A R27A K147 K129A K105A FIG.1D E161A AlanineMutantConstructs E125A E78A E69A E65A E64A D133A JUDULLUQUQU D114A D83A D71A D60A ) D46 _ _ 650. D . 0 Patent Application Publication Oct. 26 , 2017 Sheet 4 of 14 US 2017 /0306038 A1 OD650 OD650 ?não o GL OI MAB4 MAB4 1C MAB4 MAB5 Human GITR cyno GITR Human GITR Mouse GITR O Rat GITR ODFIG . 2A FIG . 2B Human GITR Ligand and MAB7 100007 Competition 9000 55h 80007000 6000 PE?hthumanMFI 5000 4000 2000 + 4nM MABZ AAAA OD650 1000 o 4nM Isotype Control hoNon 04 RmpimppiAni Amrit ? ? ö . Human Mouse Cyno Human GITR Ligand to MAB7 / Isotype GITR GITR GITR control Molar Ratio USFIG . 2C FIG . 2D Patent Application Publication Oct. 26 , 2017 Sheet 5 of 14 US 2017 /0306038 A1 OD590nm Ö62ö° WWWWH ??????????????????? BAFFRSBCMA Cova DR53 CD1373 HVEM DcR38NGFR TNFR3B 0X40 CD27 TNFR13 DR3S DR4 TACIO Cosa FAS GIINVTTTTTTT MAB4 MAB5 Control MAB FIG . 2E Patent Application Publication Oct . 26, 2017 Shet 6 of 14 US 2017 /0306038 A1 IIII? 111mm,III 0.1101000 [mAb]nM) -MAB4+XLMABSMAB5 FIG.3B [mAb]MM-MAB4+XL MABS FIG.3D [T]TWITTIT??0.001011i101001000 IIII TIT ope > P?np 64208648? Activation Fold Activation Fold ??????????????????? Concentration(nM) i101001000 ?HumanGITR-L AMAB4+XL·MAB7•MAB8 [GITR-]nM FIG.3A "dan61i101001000 FIG.3C ? ? TITm??? 765452? ? ? ? ?? ?? Activity Fold 1? Activation Fold Patent Application Publication Oct. 26 , 2017 Sheet 7 of 14 US 2017 /0306038 A1 Proliferation of CD4+ T cells Proliferation of CD8 + T cells • Proliferatingcells/uLofsample? ?©oto Proliferatingcells/plofsample 5 10 15 0 5 10 15 % GITR Ab Conjugated on Beads % GITR Ab Conjugated on Beads - IC - MAB7 + IC + MAB7 FIG . 4A FIG . 4B Secretion of IFNY [IFNy]ng/ml ?is H o6de0 5 10 15 % GITR Ab Conjugated on Beads + IC + MAB7 FIG . 4C Patent Application Publication Oct. 26 , 2017 Sheet 8 of 14 US 2017 /0306038 A1 KH H hGITR-Daudi#1F5(~5,700GITRmolecules) örg HGITR-Daudi#2011(~5,700GITRmolecules) [Ab]nM FIG.5B [Ab]nMFIG.5D öso 35007+Control3000+ MAB7 . j 0001. 0 35007+Control3000+ +MAB7 500 2500+ 500 04 250020004 >1500 1000 01ATIRE 200015001 1000 i 04A+ hGITR-Daudi#203(~4,000GITRmolecules) HGITR-Daudi#1F7(~8,700GITRmolecules) .öz FIG.5C •[AB]NM FIG.5A [Ab]nM 35007+Control30004 +MABZ azez 35007+Control3000+ MAB7 ?? 2500 15004 1000500 2500 150041000 500 >2000 220004 O4KHEH Patent Application Publication Oct. 26 , 2017 Sheet 9 of 14 US 2017 /0306038 A1 H HH %HGITR+ofCD19-CD8cells %hfctofCD19-CD8+cells 2?? o untreated-toe IgG14to NoStimolanti Ö : g CD3/CD28 untreated MAB7 (nM ) FIG .6A FIG .OB to+* %plkKtofCD19-CD8+cells KH %CD25+ofCD19-CD8cells loeto =o;$: met =o;$ 1961to 1961isto untreatedto ? Á S untreatedpeber MAB7 (nm ) MAB7 ( nM ) FIG . 6C FIG .OD Patent Application Publication Oct. 26 , 2017 Sheet 10 of 14 US 2017 /0306038 A1 MABI ZE.FIG? 90218 Vehicle doanh cells + CD8 + CD3 - CD19 of + hGITR % 16 HEH HA 14 * ISIT FIG.ZD HHHH Vehicle og DO ou cells + FOXP3 + CD4 + CD3 - CD19 of + HGITR % .MAB715mg/kg Dose 24 1 # MABT Ò 1500€Vehicle 94 Vehicle FIG.7C? § § G SEM )mm3 ( Volume Tumor Mean cells + CD8 + CD3 - CD19 of + hGITR % * 11 MAB7 FIG.7B - Vehicle 9 og 8 cells + FOXP3 + CD4 + CD3 - CD19 of + hGITR % Patent Application Publication Oct . 26, 2017 Shet 11 of14 US 2017 /0306038 A1 * * "* Vehicle? AbsoluteCD19-CD3+ ·?4:* AbsoluteCD19-CD3+CD8CD25 cellsFOXP3+CD4CD3CD19-% cells/mmtumor cels/mmstumor ….MAP74M ?? ???99 vehiceH?? MAB7. MAB7] ed999 9MAB7\ Vehicle | FIG. 8A FIG. 8B FIG . 8C -* p = 0. 052 Teff/regRatio Spotspermillioncells gegg . - ? VehicleVehicle )Me79 FIG .8E Patent Application Publication Oct. 26 , 2017 Sheet 12 of 14 US 2017 /0306038 A1 * 1 - DIA SpleenCD8PD1+ 1- ÖDTA FIG.9C 18 mlgg2a Ti ? ? 98 0 2 cells + CD8 + CD3 - CD19 of + 1 - PD % * CD8PD1+ 14- 04 Mic20mlgg2a 4000 tumor mmº per cells + PD1 + CD8 + CD3 - CD19 of Number * TumorCD8PD1+ N 14- SOTA FIG.9A -migc2a mlgG2a TiI 8 = = = p cells + CD8 + CD3 - CD19 of + PD1 % Patent Application Publication Oct. 26 , 2017 Sheet 13 of 14 US 2017 /0306038 A1 * * U co - - - - - - - - - - - TTTTT TTT B-antiRMP114+mlgg2a 71421283542 C-antiDTA–1+rlgg2aD—RMP114 FIG.10 - TTTTTTTTTTTTTTTTTTTTTTTTT Daysposttumorimplantation A—isotypecontrol + 0 study on remaining Percent juajed uogjeddy and uopjes Po 97 ' LIOT J??US JODI DI SN LIOZ/ 8E0908O IV GITR –0 + PD1 - a * $ GITR -a +PD1 - 0 Isotype + GITRGITR -- G0 ) Isotypet + GITRGITR - aO * Isotype+ PD1– 0 : 1 Isotype+ PD1 - a Isotypes de Isotypes $ $ o pom Neu on )tumor ( CD8 / PD1 ) spleen( CD8/ PD1 * - - - GITR – a + PD1 - a - GITR - a + PD1 - a * -Isotype + GITR- Isotype + GITR - - * . 1 Isotype + PD1 - a Isotype +PD1 - a FIG.11 Isotypes : : Isotypes $ ? o ?? ,) tumor( CD8 /TIM3 ) spleen ( CD8 / TIM3 ofe GITR - a + PD1 - a GITRI - a + PD1 - a : on Isotype + GITR - a Isotype + GITR - a * Isotype+ PD1 - 0 Isotype+ PD1 - a • Isotypes • : Isotypes 8 8 8 8 8 e 28? tumor CD8 / + LAG3 spleen CD8 / + LAG3 US 2017 /0306038 A1 Oct . 26 , 2017 COMPOSITIONS AND METHODS OF USE and human GITR , it is unknown whether findings seen with FOR AUGMENTED IMMUNE RESPONSE surrogate studies in mouse would translate to modification AND CANCER THERAPY of human GITR function . RELATED APPLICATIONS DESCRIPTION OF THE INVENTION [ 0001 ] This application claims priority to and the benefit [0006 ] We have identified antibodies that specifically bind of U . S . Provisional Application No. 62 / 061, 644 , filed Oct . 8 , to human glucocorticoid - induced tumor necrosis factor 2014 , U . S . Provisional Application No. 62/ 198 ,673 , filed receptor superfamily member 18 (“ GITR ” ) , wherein the Jul . 29 , 2015 , and U . S . Provisional Application No . 62/ 220 , antibodies have in vitro hGITR agonist activity when cross 764 , filed Sep . 18 , 2015 , each of which is hereby incorpo linked in vitro , and wherein the antibodies confer hGITR rated by reference in its entirety . activity in vivo and induce an elevated Teff : Treg ratio at tumor sites , resulting in inhibition of tumor progression . FIELD OF THE INVENTION Thus , the present invention provides agonist antibodies , antibody fragments , and antigen binding molecules that [ 0002 ] The present invention relates to antibodies, anti specifically bind to and promote intracellular signaling and body fragments , and antigen binding molecules that bind to or modulate immune response through targeting cells tumor necrosis factor receptor superfamily member 18 / glu expressing human GITR . In one aspect