Anti-pd-1/lag3 bispecific antibodies

Abstract

Provided herein are anti-PD-1/LAG3 bispecific antibodies and antigen-binding fragments. Also provided here are methods and uses of these antibodies and antigen-binding fragments in the treatment of cancer or infectious disease.

1 .- 41 . (canceled) 
     
     
         42 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising:
 (i) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:8, 
 (ii) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:91, 
 (iii) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:10, 
 (iv) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:3, 
 (v) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:21, and 
 (vi) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:22; and 
   (B) an anti-LAG3 antigen-binding fragment comprising:
 (i) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:112, 
 (ii) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:113, 
 (iii) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:114, 
 (iv) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:115, 
 (v) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:116, and 
 (vi) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:117. 
   
     
     
         43 . The method of  claim 42 , wherein the anti-PD-1 heavy chain variable region CDR2 comprises the amino acid sequence of SEQ ID NO:86. 
     
     
         44 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising
 (A) an anti-PD-1 antigen-binding fragment comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:92, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:14, 17, 20, 24, 28, 31, or 34; and   (B) an anti-LAG3 antigen-binding fragment comprising an anti-LAG3 heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:99.   
     
     
         45 . The method of  claim 44 , wherein the anti-PD-1 antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:92, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:20. 
     
     
         46 . The method of  claim 44 , wherein the anti-PD-1 antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:85, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:20. 
     
     
         47 . The method of  claim 45 , comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:92, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and CH3 mutations 350V, 351Y, 405A, and 407V, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:20, and a kappa constant region comprising Cκ mutations 124R and 178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and an IgG1 constant region comprising CH1 mutation 181K, and CH3 mutations 350V, 366L, 392L, and 394W, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the mutations are in EU numbering.   
     
     
         48 . The method of  claim 45 , comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:92, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and CH3 mutations 350V, 366L, 392L, and 394W, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:20, and a kappa constant region comprising Cκ mutations 124R and 178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97 and an IgG1 constant region comprising CH1 mutation 181K, and CH3 mutations 350V, 351Y, 405A, and 407V, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the mutations are in EU numbering.   
     
     
         49 . The method of  claim 45 , comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:92, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:20, and a kappa constant region comprising Cκ mutations 124R and 178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and an IgG1 constant region comprising CH1 mutation 181K, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the IgG1 heavy chain constant regions of the anti-PD-1 and anti-LAG3 antigen-binding fragments further comprise pairs of CH3 mutations selected from the group consisting of: 351Y/405A/407V and 366I/392M/394W; 351Y/405A/407V and 366L/392L/394W; 351Y/405A/407V and 366L/392M/394W, and   wherein the mutations are in EU numbering.   
     
     
         50 . The method of  claim 44 , comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:92, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and CH3 mutations 350V, 351Y, 405A, and 407V, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:14, 17, 20, 24, 28, 31, or 34, and a kappa constant region comprising Cκ mutations 124R and 178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and an IgG1 constant region comprising CH1 mutation 181K, and CH3 mutations 350V, 366L, 392L, and 394W, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the mutations are in EU numbering.   
     
     
         51 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 heavy chain comprising the amino acid sequence of SEQ ID NO:102, and a light chain comprising the amino acid sequence of SEQ ID NO:103, and   (B) an anti-LAG3 heavy chain comprising the amino acid sequence of SEQ ID NO:96, and a light chain comprising the amino acid sequence of SEQ ID NO:98.   
     
     
         52 . The method of  claim 51 , wherein the antibody or antigen-binding fragment thereof comprises a glycosylation pattern characteristic of expression by a CHO cell. 
     
     
         53 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD 1 antigen-binding fragment comprising:
 (i) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:8, 
 (ii) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:86, 
 (iii) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:10 or 41, 
 (iv) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:3, 
 (v) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:4, and 
 (vi) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:5; and 
   (B) an anti-LAG3 antigen-binding fragment comprising:
 (i) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:112, 
 (ii) a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:113, 
 (iii) a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:114, 
 (iv) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO:115, 
 (v) a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO:116, and 
 (vi) a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO:117. 
   
     
     
         54 . The method of  claim 53 , wherein
 (A) the anti-PD-1 antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:95, and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:76; and   (B) the anti-LAG3 antigen-binding fragment comprises an anti-LAG3 heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and light chain variable region comprising the amino acid sequence of SEQ ID NO:99.   
     
     
         55 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:93, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and CH3 mutations 350V, 351Y, 405A, and 407V, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:76, and a kappa constant region comprising Cκ mutations Q124R and T178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and an IgG1 constant region comprising CH1 mutation 181K, and CH3 mutations 350V, 366L, 392L, and 394W, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the mutations are in EU numbering.   
     
     
         56 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:93, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and CH3 mutations 350V, 366L, 392L, and 394W, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:76, and a kappa constant region comprising Cκ mutations 124R and 178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and an IgG1 constant region comprising CH1 mutation 181K, and CH3 mutations 350V, 351Y, 405A, and 407V, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the mutations are in EU numbering.   
     
     
         57 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:93, and an IgG1 constant region comprising CH1 mutations 145E, 147T, 175E, and 183L, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:76, and a kappa constant region comprising Cκ mutations 124R and 178R; and   (B) an anti-LAG3 antigen-binding fragment comprising (i) a heavy chain comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:97, and an IgG1 constant region comprising CH1 mutation 181K, and (ii) a light chain comprising a light chain variable region comprising the amino acid sequence of SEQ ID NO:99, and a kappa constant region comprising Cκ mutations 124E, 131T, 178Y, and 180E;   wherein the IgG1 heavy chain constant regions of the anti-PD-1 and anti-LAG3 antigen-binding fragments further comprise pairs of CH3 mutations selected from the group consisting of: 351Y/405A/407V and 366I/392M/394W; 351Y/405A/407V and 366L/392L/394W; and 351Y/405A/407V and 366L/392M/394W, and   wherein the mutations are in EU numbering.   
     
     
         58 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 heavy chain comprising the amino acid sequence of SEQ ID NO:101, and a light chain comprising the amino acid sequence of SEQ ID NO:100, and   (B) an anti-LAG3 heavy chain comprising the amino acid sequence of SEQ ID NO:96, and a light chain comprising the amino acid sequence of SEQ ID NO:98.   
     
     
         59 . A method of treating cancer or an infection or infectious disease in a human subject, comprising administering to the subject an effective amount of an anti-PD-1/LAG-3 bispecific antibody, comprising:
 (A) an anti-PD-1 heavy chain variable region comprising the amino acid sequence of SEQ ID NO:109, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:111, and   (B) an anti-LAG3 heavy chain variable region comprising the amino acid sequence of SEQ ID NO:105, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:107.   
     
     
         60 . The method of  claim 59 , comprising:
 (A) an anti-PD-1 heavy chain comprising the amino acid sequence of SEQ ID NO:108, and a light chain comprising the amino acid sequence of SEQ ID NO:110, and   (B) an anti-LAG3 heavy chain comprising the amino acid sequence of SEQ ID NO:104, and a light chain comprising the amino acid sequence of SEQ ID NO:106.   
     
     
         61 . The method of  claim 42 , wherein the further therapeutic agent is selected from the group consisting of: (i) an anti-TIGIT antibody or an antigen-binding fragment thereof; (ii) an anti-VISTA antibody or an antigen-binding fragment thereof; (iii) an anti-BTLA antibody or an antigen-binding fragment thereof; (iv) an anti-TIM3 antibody or an antigen-binding fragment thereof; (v) an anti-CTLA4 antibody or an antigen-binding fragment thereof; (vi) an anti-HVEM antibody or an antigen-binding fragment thereof; (vii) an anti-CD70 antibody or an antigen-binding fragment thereof; (viii) an anti-OX40 antibody or an antigen-binding fragment thereof; (ix) an anti-CD28 antibody or an antigen-binding fragment thereof; (x) an anti-PDL1 antibody or an antigen-binding fragment thereof; (xi) an anti-PDL2 antibody or an antigen-binding fragment thereof; (xii) an anti-GITR antibody or an antigen-binding fragment thereof; (xiii) an anti-ICOS antibody or an antigen-binding fragment thereof; (xiv) an anti-SIRPα antibody or an antigen-binding fragment thereof; (xv) an anti-ILT2 antibody or an antigen-binding fragment thereof; (xvi) an anti-ILT3 antibody or an antigen-binding fragment thereof; (xvii) an anti-ILT4 antibody or an antigen-binding fragment thereof; (xviii) an anti-ILT5 antibody or an antigen-binding fragment thereof; (xix) an anti-4-1BB antibody or an antigen-binding fragment thereof; (xx) an anti-NK2GA antibody or an antigen-binding fragment thereof; (xxi) an anti-NK2GC antibody or an antigen-binding fragment thereof; (xxii) an anti-NK2GE antibody or an antigen-binding fragment thereof; (xxiii) an anti-TSLP antibody or an antigen-binding fragment thereof; (xxiv) an anti-IL10 antibody or an antigen-binding fragment thereof; (xxv) a STING agonist; (xxvi) a CXCR2 antagonist; and (xxvii) a PARP inhibitor.