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(12) Patent Application Publication (10) Pub. No.: US 2016/0319019 A1 Amirina Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2016/0319019 A1 Amirina Et Al US 201603.19019A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0319019 A1 Amirina et al. (43) Pub. Date: Nov. 3, 2016 (54) ANTI-PD-1 ANTIBODIES (22) Filed: May 11, 2016 (71) Applicant: Enumeral Biomedical Holdings, Inc., Related U.S. Application Data Cambridge, MA (US) (62) Division of application No. 14/975,769, filed on Dec. 19, 2015. (72) Inventors: Najmia Amirina, Cambridge, MA (60) Provisional application No. 62/095.675, filed on Dec. (US); Hareesh Chamarthi, Allston, 22, 2014, provisional application No. 62/220,199, MA (US); Maria Isabel Chiu, Newton filed on Sep. 17, 2015, provisional application No. Centre, MA (US); Daniel Doty, 62/251,082, filed on Nov. 4, 2015, provisional appli Arlington, MA (US); Bin Feng, cation No. 62/261,118, filed on Nov. 30, 2015. Newton, MA (US); Aleksander Jonca, Publication Classification Boston, MA (US); Thomas McQuade, Cambridge, MA (US); Anhco Nguyen, (51) Int. Cl. Needham, MA (US); Sheila C07K 6/28 (2006.01) Ranganath, Arlington, MA (US); Hans (52) U.S. Cl. Albert Felix Scheuplein, Arlington, CPC ..... C07K 16/2803 (2013.01); C07K 231 7/567 MA (US); Vikki A. Spaulding, Acton, (2013.01); C07K 23.17/34 (2013.01); A61 K MA (US); Lei Wang, Braintree, MA 2039/507 (2013.01) (US); Jennifer Watkins-Yoon, Brighton, MA (US) (57) ABSTRACT Antibodies that bind to programmed cell death protein 1 (PD-1), compositions comprising Such antibodies, and (21) Appl. No.:y x- - - 15/152,1929 methods of making and using Such antibodies are disclosed. Patent Application Publication Nov. 3, 2016 Sheet 3 of 27 US 2016/0319019 A1 s's is 5 w c. x. 9."OIH N were (uu Ogiy)CO Patent Application Publication Nov. 3, 2016 Sheet 4 of 27 US 2016/0319019 A1 sasaasaaaaaaa. ~~~~~~~~~~&~~~~~~~~~~ ~~~~~~~~~~~*~~~~~~~~~~~ vs. xxxxxx Sasa-sa-sass-as-s-s-s-s-s-s was saw sassssssssssssssssssass-s-s-s-s-s-s-s-s-as-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s-s www.www. O O O O O O O O 06 ox n S y n s eNISOd ANIAueoued }} } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } * } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Patent Application Publication Nov. 3, 2016 Sheet 5 of 27 US 2016/0319019 A1 eAI)SO 90 Patent Application Publication Nov. 3, 2016 Sheet 6 of 27 US 2016/0319019 A1 GN5 (N.: V. , V. , O., O.. 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Od (peef v00)+GOOJO % 23 Patent Application Publication Nov. 3, 2016 Sheet 10 of 27 US 2016/0319019 A1 FIG 9A COS 38 sit COS 3. isotype Control a 3.3 388): 20 33S s S is R&C8 8 s3A is : O WXW WOW WWWWWWwww WWW 8xciseesis............. is . : Antibody Concentiations FIG 9B R 4, Ea. af 3.S isotype Control 3. 383 & & V- s 43 s S SA: O {, --firessessessississisterest:- i. 3. - 18 Antibody Concentration (n& Patent Application Publication Nov. 3, 2016 Sheet 11 of 27 US 2016/0319019 A1 FIG 9C (28 ce- a-28 3 O -- isotype Cotto X5 are 338. { -- a 3. s 8- at a 34&CS m -&- 238áO & O assessesseekeepees O{}} { O. t O O Antibody Cor:certation (rfi FIG 9D CA4 3.S -- 8; it...O.A4: 3. -- isotype Cotroi e are 388 a 20 -- (3: s S - w8- 244C& career {} - a8A: s : s& . {{ 8. Artibody Cocentration (rh) Patent Application Publication Nov. 3, 2016 Sheet 14 of 27 US 2016/0319019 A1 FIG 11 A 88x8 88: 888-&ies: 888 $88-8- 83.3 : . 33 S && $38 &&. FIG 11B Patent Application Publication Nov. 3, 2016 Sheet 15 of 27 US 2016/0319019 A1 &ays six}s $88-&iska Fiat 388-&- 88.8 : & 88 & 83 &SS-S. FIG 11 C 8888 First 388-88x8 Fix. $88-&- &: : S.S.S A & S333 S&S FIG 11D Patent Application Publication Nov. 3, 2016 Sheet 16 of 27 US 2016/0319019 A1 FIG. 12 Patent Application Publication Nov. 3, 2016 Sheet 17 of 27 US 2016/0319019 A1 | FIG. 13 Patent Application Publication Nov. 3, 2016 Sheet 18 of 27 US 2016/0319019 A1 w re 8E.84 ER is ; ASé8. i-Nd its anpu ps: Patent Application Publication Nov. 3, 2016 Sheet 19 of 27 US 2016/0319019 A1 15000 10000 5000 FIG 15A 400 23 OOOO 1 O O FIG 15B Patent Application Publication Nov. 3, 2016 Sheet 20 of 27 US 2016/0319019 A1 150 1 O O 5 O FIG 15C 800 6 O O 4. OO 200 O 2 N N N S$ S$ S$ Q Q Q vS N vS SN SS. Wcy O S S ?:W C OS c We WS FIG 15D Patent Application Publication Nov. 3, 2016 Sheet 21 of 27 US 2016/0319019 A1 30 2O 10 FIG 15E 100 50 O 2 FIG 15F Patent Application Publication Nov. 3, 2016 Sheet 22 of 27 US 2016/0319019 A1 40000 3OOOO 20000 1OOOO FIG 1.6A 100 80 60 40 20 FIG. 16B Patent Application Publication Nov. 3, 2016 Sheet 23 of 27 US 2016/0319019 A1 FIG. 16C 1500 1OOO 500 FIG. 16D Patent Application Publication Nov. 3, 2016 Sheet 24 of 27 US 2016/0319019 A1 200 5 O FIG. 16E FIG. 16F Patent Application Publication Nov. 3, 2016 Sheet 25 of 27 US 2016/0319019 A1 25 -- Group 1: Vehicle -- Group 2: 388D4-3 & Group 3: 244C8-2 1000 -X. Group 4. Pembroizumab -(- Group 5: Pembroizumab + 244C8-2 S. S 25 FIG 17A Patent Application Publication Nov. 3, 2016 Sheet 26 of 27 US 2016/0319019 A1 \\\\\\\\\\\\\\\\ 'YYYYYYYYYYYYYYYYYXXXXXXXXXXXXXXXXXXXXX W W sexyx pay . SES ES s S. eunOWJOun Patent Application Publication Nov. 3, 2016 Sheet 27 of 27 US 2016/0319019 A1 Z C)LI"OIH ×× S.S. SS SS & e6eueoue eunOWJOun US 2016/03 19019 A1 Nov. 3, 2016 ANT-PD-1. ANTIBODES SUMMARY OF THE INVENTION 0008. The present invention provides antibodies that bind RELATED APPLICATIONS to PD-1. In some embodiments, the invention provides an 0001. This application is a divisional of U.S. application isolated antibody that binds to PD-1, comprising a heavy Ser. No. 14/975,769, filed Dec. 19, 2015, which claims the chain variable region (HCVR) selected from the group benefit of U.S. Provisional Application No. 62/095.675, filed consisting of SEQID NOs: 1-26 and/or a light chain variable on Dec. 22, 2014, U.S. Provisional Application No. 62/220, region (LCVR) selected from the group consisting of SEQ 199, filed on Sep. 17, 2015, U.S. Provisional Application ID NOS: 27-53. The invention also provides an isolated No. 62/251,082, filed on Nov. 4, 2015, and U.S. Provisional antibody that binds to PD-1 and competitively inhibits the Application No. 62/261,118, filed on Nov. 30, 2015. The binding of any of the antibodies disclosed herein to PD-1. entire teachings of the above applications are incorporated 0009. In some embodiments, the invention also provides herein by reference. an isolated antibody that binds to PD-1, comprising a HCVR selected from the group consisting of SEQ ID NOs: 85-90 INCORPORATION BY REFERENCE OF and/or a LCVR selected from the group consisting of SEQ MATERIAL IN ASCII TEXT FILE ID NOS: 91-96. 0010. The invention further provides an isolated antibody 0002 This application incorporates by reference the that binds to PD-1, wherein the antibody binds to a sequence Sequence Listing contained in the following ASCII text file in PD-1 selected from the group consisting of SEQID NOs: being submitted concurrently herewith: 54-84. 0003 a) File name: 5091 1000006SEQUENCELIST 0011. The antibodies can be used as therapeutic agents. ING..txt; created May 11, 2016, 74 KB in size. For use as therapeutic agents, the antibodies disclosed herein can be engineered, e.g., humanized, to reduce or eliminate BACKGROUND OF THE INVENTION serum sickness or an undesired immune response when administered to a human patient. Also disclosed are methods 0004 Modulation of the mammalian adaptive immune of treating diseases and disorders in which the PD-1 signal response (immunomodulation) is a useful therapeutic ing pathway plays a significant role (“PD-1-mediated dis approach for various diseases and disorders. One way to eases and disorders'). achieve Such immunomodulation is to intervene at one or 0012. The present invention includes the surprising dis more immune checkpoints, e.g., the Programmed Death-1 covery that contacting human T cells with an effective (PD-1) checkpoint. The natural function of immune check amount of an anti-PD-1 antibody that competitively inhibits points is to Suppress the immune response, as necessary, to binding of PD-L1 or PD-L2 to PD-1 expressed on the prevent immune damage to normal tissue. Depending on the surface of T cells, and an effective amount of an anti-PD-1 disease or disorder, it may be desirable to upregulate or antibody that does not competitively inhibit binding of downregulate the immune response. Tumor cells that display PD-L1 or PD-L2 to PD-1 expressed on the surface of the T non-self-antigens can evade immune attack by secreting cells increases T cell effector function to a greater extent cytokines or ligands that activate immune checkpoints.
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