Antibodies and methods of use thereof

Abstract

The present disclosure provides multispecific (e.g., bispecific) antibodies that specifically bind to human GITR and/or human OX40 as well as compositions comprising such antibodies. In a specific aspect, the multispecific antibodies specifically bind to human GITR and OX40 and modulate GITR and/or OX40 activity, e.g., enhance, activate, or induce GITR and/or OX40 activity, or reduce, deactivate, or inhibit GITR and/or OX40 activity. The present disclosure also provides methods for treating disorders, such as cancer, by administering a multispecific antibody that specifically binds to human GITR and/or OX40 and modulates GITR and/or OX40 activity, e.g., enhances, activates, or induces GITR and/or OX40 activity. Also provided are methods for treating autoimmune or inflammatory diseases or disorders, by administering a multispecific antibody that specifically binds to human GITR and/or OX40 and modulates GITR and/or OX40 activity, e.g., reduces, deactivates, or inhibits GITR and/or OX40 activity.

1 .- 188 . (canceled) 
     
     
         189 . A method of modulating an immune response in a subject in need thereof, the method comprising administering to the subject an effective amount of a multispecific antibody comprising a first antigen-binding region that specifically binds human OX40 and a second antigen-binding region that specifically binds human GITR, wherein:
 a. the first antigen-binding region comprises a first heavy chain comprising CDRs VH-CDR1, VH-CDR2, and VH-CDR3, and a first light chain comprising CDRs VL-CDR1, VL-CDR2, and VL-CDR3, wherein the first heavy chain comprises the VH-CDR1, VH-CDR2, and VH-CDR3 amino acid sequences of the VH amino acid sequence of SEQ ID NO: 54, and the first light chain comprises the VL-CDR1, VL-CDR2, and VL-CDR3 amino acid sequences of the VL amino acid sequence of SEQ ID NO: 55 and   b. the second antigen-binding region comprises a second antibody heavy chain comprising CDRs VH-CDR1, VH-CDR2, and VH-CDR3 and a second antibody light chain comprising CDRs VL-CDR1, VL-CDR2, and VL-CDR3, wherein:
 i. the VH-CDR1 comprises the amino acid sequence of X 1 YX 2 MX 3  (SEQ ID NO:87), wherein X 1  is D, E or G; X 2  is A or V; and X 3  is Y or H; 
 ii. the VH-CDR2 comprises the amino acid sequence of X 1 IX 2 TX 3 SGX 4 X 5 X 6 YNQKFX 7 X 8 (SEQ ID NO:88), wherein X 1  is V or L; X 2  is R, K or Q; X 3  is Y or F; X 4  is D, E or G; X 5  is V or L; X 6  is T or S; X 7  is K, R or Q; and X 8  is D, E or G; 
 iii. the VH-CDR3 comprises the amino acid sequence of SEQ ID NO:3; 
 iv. the VL-CDR1 comprises the amino acid sequence of KSSQSLLNSX 1 NQKNYLX 2  (SEQ ID NO:90), wherein X 1  is G or S; and X 2  is T or S; 
   v. the VL-CDR2 comprises the amino acid sequence of SEQ ID NO:5; and   vi. the VL-CDR3 comprises the amino acid sequence of QNX 1 YSX 2 PYT (SEQ ID NO:92), wherein X 1  is D or E; and X 2  is Y, F or S.   
     
     
         190 . The method of  claim 189 , wherein:
 a. the VH-CDR1 of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:47;   b. the VH-CDR2 of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:48;   c. the VH-CDR3 of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:49;   d. the VL-CDR1 of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:50;   e. the VL-CDR2 of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:51; and   f. the VL-CDR3 of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:52 or SEQ ID NO:53.   
     
     
         191 . The method of  claim 189 , wherein the first antigen-binding region comprises a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:52. 
     
     
         192 . The method of  claim 189 , wherein the first antigen-binding region comprises a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:53. 
     
     
         193 . The method of  claim 189 , wherein:
 a. (i) the VH-CDR1 of the second antigen-binding region comprises the amino acid sequence of X 1 YAMX 2  (SEQ ID NO:1), wherein X 1  is D, G, or E; and X 2  is Y or H; (ii) the VH-CDR2 of the second antigen-binding region comprises the amino acid sequence of X 1 IRTYSGX 2 VX 3 YNQKFX 4 X 5  (SEQ ID NO:2), wherein X 1  is V or L; X 2  is D or G; X 3  is T or S; X 4  is K, R, or Q; and X 5  is D, E, or G; (iii) the VH-CDR3 of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:3; (iv) the VL-CDR1 of the second antigen-binding region comprises the amino acid sequence of KSSQSLLNSX 1 NQKNYLT (SEQ ID NO:4), wherein X 1  is G or S; (v) the VL-CDR2 of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:5; and (vi) the VL-CDR3 of the second antigen-binding region comprises the amino acid sequence of QNX 1 YSX 2 PYT (SEQ ID NO:6), wherein X 1  is D or E; and X 2  is Y or F;   b. the VH-CDR1 of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:7, 8, or 9;   c. the VH-CDR2 of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:10, 11, 12, or 13;   d. the VL-CDR1 of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:14 or 15;   e. the VL-CDR3 of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:16 or 17;   f. the VH-CDR1, the VH-CDR2, and the VH-CDR3 of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs:7, 10, and 3; 8, 11, and 3; 9, 12, and 3; or 9, 13, and 3, respectively; and/or the VL-CDR1, the VL-CDR2, and the VL-CDR3 of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs:14, 5, and 16; or 15, 5, and 17, respectively; or   g. the VH-CDR1, the VH-CDR2, the VH-CDR3, the VL-CDR1, the VL-CDR2, and the VL-CDR3 of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs:7, 10, 3, 14, 5, and 16, respectively.   
     
     
         194 . The method of  claim 189 , wherein:
 a. the VH of the first antigen-binding region comprises an amino acid sequence that is at least 75% identical to the amino acid sequence of SEQ ID NO:54;   b. the VH of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:54;   c. the VH of the first antigen-binding region comprises an amino acid sequence derived from a human IGHV3-73 germline sequence;   d. the VL of the first antigen-binding region comprises an amino acid sequence at least 75% identical to the amino acid sequence of SEQ ID NO:55 or SEQ ID NO:56;   e. the VL of the first antigen-binding region comprises the amino acid sequence of SEQ ID NO:55 or 56;   f. the VL of the first antigen-binding region comprises an amino acid sequence derived from a human IGKV2-28 germline sequence;   g. the VH and the VL of the first antigen-binding region comprise the amino acid sequences of SEQ ID NOs:54 and 55; or 54 and 56, respectively; or   h. the VH of the first antigen-binding region comprises an amino acid sequence derived from a human IGHV3-73 germline sequence and a VL comprising an amino acid sequence derived from a human IGKV2-28 germline sequence.   
     
     
         195 . The method of  claim 189 , wherein:
 a. the VH of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:25;   b. the VH of the second antigen-binding region comprises an amino acid sequence at least 75% identical to the amino acid sequence of SEQ ID NO:18, 20, 22, or 24;   c. the VH of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:18, 20, 22, or 24;   d. the VH of the second antigen-binding region comprises an amino acid sequence derived from a human IGHV1-2 germline sequence;   e. the VL of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:26;   f. the VL of the second antigen-binding region comprises an amino acid sequence at least 75% identical to the amino acid sequence of SEQ ID NO:19, 21, or 23;   g. the VL of the second antigen-binding region comprises the amino acid sequence of SEQ ID NO:19, 21, or 23;   h. the VL of the second antigen-binding region comprises an amino acid sequence derived from a human IGKV4-1 germline sequence;   i. the VH and the VL of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs:18 and 19; 20 and 21; 22 and 23; or 24 and 23, respectively; or   j. the VH of the second antigen-binding region comprises an amino acid sequence derived from a human IGHV1-2 germline sequence and a VL comprising an amino acid sequence derived from a human IGKV4-1 germline sequence.   
     
     
         196 . The method of  claim 189 , wherein:
 a. the first heavy chain comprises the amino acid sequence of SEQ ID NO:59, 60, 66, 118, 119, or 125;   b. the first light chain comprises the amino acid sequence of SEQ ID NO:67, 68, 69, or 70;   c. the first heavy chain comprises the amino acid sequence of SEQ ID NO:59 and the first light chain comprises the amino acid sequence of SEQ ID NO:67 or 69; or   d. the first heavy chain comprises the amino acid sequence of SEQ ID NO:118 and the first light chain comprises the amino acid sequence of SEQ ID NO:67 or 69.   
     
     
         197 . The method of  claim 189 , wherein:
 a. the second heavy chain comprises the amino acid sequence of SEQ ID NO:29, 30, 36, 74, 74, or 81;   b. the second light chain comprises the amino acid sequence of SEQ ID NO:37 or 38;   c. the second heavy chain comprises the amino acid sequence of SEQ ID NO:29 and the second light chain comprises the amino acid sequence of SEQ ID NO:37;   d. the second heavy chain comprises the amino acid sequence of SEQ ID NO:74 and the second light chain comprises the amino acid sequence of SEQ ID NO:37;   e. the second heavy chain comprises the amino acid sequence of SEQ ID NO:30 and the second light chain comprises the amino acid sequence of SEQ ID NO:37; or   f. the second heavy chain comprises the amino acid sequence of SEQ ID NO:75 and the second light chain comprises the amino acid sequence of SEQ ID NO:37.   
     
     
         198 . The method of  claim 189 , wherein the first heavy chain and/or the second heavy chain comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 . 
     
     
         199 . The method of  claim 198 , wherein the first heavy chain and/or the second heavy chain comprises an IgG 1  heavy chain constant region. 
     
     
         200 . The method of  claim 199 , wherein:
 a. the first heavy chain comprises serine, alanine, and valine at amino acid positions 366, 368, and 407, respectively, and the second heavy chain comprises tryptophan at amino acid position 366; or   b. the first heavy chain comprises tryptophan at amino acid position 366, and the second heavy chain comprises serine, alanine, and valine at amino acid positions 366, 368, and 407, respectively, numbered according to the EU numbering system.   
     
     
         201 . The method of  claim 200 , wherein the first heavy chain and/or the second heavy chain comprises aspartate, leucine, and glutamate at amino acid positions 239, 330, and 332, respectively, numbered according to the EU numbering system. 
     
     
         202 . The method of  claim 189 , wherein the first light chain and/or the second light chain comprises a human kappa light chain constant region or a human lambda light chain constant region. 
     
     
         203 . The method of  claim 199 , wherein:
 a. the first heavy chain comprises the amino acid sequence of SEQ ID NO:54, and the first light chain comprises the amino acid sequence of SEQ ID NO:67; and   b. the second heavy chain comprises the amino acid sequence of SEQ ID NO:18, and the second light chain comprises the amino acid sequence of SEQ ID NO:37.   
     
     
         204 . The method of  claim 189 , wherein the first heavy chain comprises aspartate, leucine, and glutamate at amino acid positions 239, 330, and 332, respectively, numbered according to the EU numbering system. 
     
     
         205 . The method of  claim 189 , wherein the second heavy chain comprises aspartate, leucine, and glutamate at amino acid positions 239, 330, and 332, respectively, numbered according to the EU numbering system. 
     
     
         206 . The method of  claim 189 , wherein the first heavy chain and the second heavy chain comprise aspartate, leucine, and glutamate at amino acid positions 239, 330, and 332, respectively, numbered according to the EU numbering system. 
     
     
         207 . The method of  claim 198 , wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 126. 
     
     
         208 . The method of  claim 189 , wherein the immune response is enhanced or induced in the subject. 
     
     
         209 . The method of  claim 189 , wherein the subject has cancer. 
     
     
         210 . The method of  claim 209 , wherein the cancer is selected from the group consisting of melanoma, renal cancer, prostate cancer, colon cancer, and lung cancer. 
     
     
         211 . The method of  claim 209 , further comprising administering to the subject an inhibitor of indoleamine-2,3-dioxygenase (IDO). 
     
     
         212 . The method of  claim 211 , wherein the inhibitor is selected from the group consisting of epacadostat, F001287, indoximod, and NLG919. 
     
     
         213 . The method of  claim 209 , further comprising administering to the subject a vaccine. 
     
     
         214 . The method of  claim 213 , wherein the vaccine comprises a heat shock protein peptide complex (HSPPC) comprising a heat shock protein complexed with an antigenic peptide. 
     
     
         215 . The method of  claim 214 , wherein the heat shock protein is hsp70 or hsc70 and is complexed with a tumor-associated antigenic peptide. 
     
     
         216 . The method of  claim 215 , wherein the heat shock protein is gp96 and is complexed with a tumor-associated antigenic peptide, wherein the HSPPC is derived from a tumor obtained from the subject. 
     
     
         217 . The method of  claim 217 , further comprising administering to the subject a checkpoint targeting agent. 
     
     
         218 . The method of  claim 217 , wherein the checkpoint targeting agent is selected from the group consisting of an antagonist anti-PD-1 antibody, an antagonist anti-PD-L1 antibody, an antagonist anti-PD-L2 antibody, an antagonist anti-CTLA-4 antibody, an antagonist anti-TIM-3 antibody, an antagonist anti-LAG-3 antibody, an antagonist anti-CEACAM1 antibody, an agonist anti-GITR antibody, and an agonist anti-OX40 antibody. 
     
     
         219 . The method of  claim 189 , wherein the subject has an infectious disease. 
     
     
         220 . The method of  claim 189 , wherein immune response is reduced or inhibited. 
     
     
         221 . The method of  claim 189 , wherein the subject has an autoimmune or inflammatory disease or disorder. 
     
     
         222 . The method of  claim 221 , wherein the autoimmune or inflammatory disease or disorder is selected from the group consisting of transplant rejection, graft-versus-host disease, vasculitis, asthma, rheumatoid arthritis, dermatitis, inflammatory bowel disease, uveitis, lupus, colitis, diabetes, multiple sclerosis, and airway inflammation. 
     
     
         223 . The method of  claim 189 , wherein the subject is human.