US2016222117A1PendingUtilityA1

Nucleic acid encoding anti-pd-l1 antibodies

Assignee: GENENTECH INCPriority: Dec 9, 2008Filed: Sep 4, 2015Published: Aug 4, 2016
Est. expiryDec 9, 2028(~2.4 yrs left)· nominal 20-yr term from priority
A61P 31/10A61P 43/00A61P 33/00A61P 35/00A61P 37/02A61P 31/12A61P 33/02A61P 31/00A61P 37/00A61P 31/04A61P 37/04C07K 16/1145C07K 16/2827C07K 16/30A61K 39/3955C07K 16/22C07K 16/3046C07K 2317/565C07K 2317/74C07K 2317/71A61K 2039/505C07K 2317/56C07K 2317/73A61K 39/39558C07K 2317/52C07K 2317/24C07K 2317/92A61K 2039/507C07K 2317/76C07K 2317/14A61K 31/7068C07K 16/28A61K 45/06C07K 2317/567A61K 39/00Y02A50/30A61K 2300/00
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Claims

Abstract

The present application relates to nucleic acid encoding anti-PD-L1 antibodies, which have therapeutic use to enhance T-cell function to upregulate cell-mediated immune responses and for the treatment of T cell dysfunctional disorders, including infection (e.g., acute and chronic) and tumor immunity.

Claims

exact text as granted — not AI-modified
1 . An isolated nucleic acid encoding a heavy chain variable region polypeptide that specifically binds to PD-L1 comprising an HVR-H1, HVR-H2 and HVR-H3 sequence, wherein:
 (a) the HVR-H1 sequence is GFTFSX 1 SWIH (SEQ ID NO:1);   (b) the HVR-H2 sequence is AWIX 2 PYGGSX 3 YYADSVKG (SEQ ID NO:2);   (c) the HVR-H3 sequence is RHWPGGFDY (SEQ ID NO:3);   further wherein: X 1  is D or G; X 2  is S or L; X 3  is T or S.   
     
     
         2 . The nucleic acid of  claim 1  wherein X 1  is D; X 2  is S and X 3  is T. 
     
     
         3 . The nucleic acid of  claim 1  further comprising variable region heavy chain framework sequences juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4). 
     
     
         4 . The nucleic acid of  claim 3  wherein the framework sequences are human. 
     
     
         5 . The nucleic acid of  claim 4  wherein the framework sequences are VH subgroup III consensus framework. 
     
     
         6 . The nucleic acid of  claim 5  wherein one or more of the framework sequences is the following: 
       
         
           
                 
                 
               
                     
                   HC-FR1 is 
                 
                 
               
                   (SEQ ID NO: 4) 
                 
                 
                 
               
                     
                   EVQLVESGGGLVQPGGSLRLSCAAS 
                 
                     
                     
                 
                     
                   HC-FR2 is 
                 
                 
               
                   (SEQ ID NO: 5) 
                 
                 
                 
               
                     
                   WVRQAPGKGLEWV 
                 
                     
                     
                 
                     
                   HC-FR3 is 
                 
                 
               
                   (SEQ ID NO: 6) 
                 
                 
                 
               
                     
                   RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR 
                 
                     
                     
                 
                     
                   HC-FR4 is 
                 
                 
               
                   (SEQ ID NO: 7) 
                 
                 
                 
               
                     
                   WGQGTLVTVSA. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         7 . The nucleic acid of  claim 1 , further comprising a second nucleic acid encoding a variable region light chain polypeptide comprising an HVR-L1, HVR-L2 and HVR-L3, wherein:
 (a) the HVR-L1 sequence is RASQX 4 X 5 X 6 TX 7 X 8 A (SEQ ID NOs:8);   (b) the HVR-L2 sequence is SASX 9 LX 10 S, and (SEQ ID NOs:9);   (c) the HVR-L3 sequence is QQX 11 X 12 X 13 X 14 PX 15 T (SEQ ID NOs:10);   further wherein: X 4  is D or V; X 5  is V or I; X 6  is S or N; X 7  is A or F; X 8  is V or L; X 9  is F or T; X 10  is Y or A; X 11  is Y, G, F, or S; X 12  is L, Y, F or W; X 13  is Y, N, A, T, G, F or I; X 14  is H, V, P, T or I; X 15  is A, W, R, P or T.   
     
     
         8 . The nucleic acid of  claim 7  wherein X 4  is D; X 5  is V; X 6  is S; X 7  is A; X 8  is V; X 9  is F; X 10  is Y; X 11  is Y; X 12  is L; X 13  is Y; X 14  is H; X 15  is A. 
     
     
         9 . The nucleic acid of  claim 7  further comprising variable region light chain framework sequences juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L 1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). 
     
     
         10 . The nucleic acid of  claim 9  wherein the framework sequences are human. 
     
     
         11 . The nucleic acid of  claim 10  wherein the framework sequences are VL kappa I consensus framework. 
     
     
         12 . The nucleic acid of  claim 11  wherein one or more of the framework sequences is the following: 
       
         
           
                 
                 
               
                     
                   LC-FR1 is 
                 
                 
               
                   (SEQ ID NO: 11) 
                 
                 
                 
               
                     
                   DIQMTQSPSSLSASVGDRVTITC; 
                 
                     
                     
                 
                     
                   LC-FR2 is 
                 
                 
               
                   (SEQ ID NO: 12) 
                 
                 
                 
               
                     
                   WYQQKPGKAPKLLIY; 
                 
                     
                     
                 
                     
                   LC-FR3 is 
                 
                 
               
                   (SEQ ID NO: 13) 
                 
                 
                 
               
                     
                   GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC; 
                 
                     
                     
                 
                     
                   LC-FR4 is 
                 
                 
               
                   (SEQ ID NO: 14) 
                 
                 
                 
               
                     
                   FGQGTKVEIKR. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         13 - 24 . (canceled) 
     
     
         25 . An isolated nucleic acid encoding a light chain or a heavy chain variable region sequence of an anti-PD-L1 antibody or antigen binding fragment, wherein:
 (a) the heavy chain or antigen binding fragment thereof further comprises and HVR-H1, HVR-H2 and an HVR-H3 sequence having at least 85% sequence identity to GFTFSDSWIH (SEQ ID NO:15), AWISPYGGSTYYADSVKG (SEQ ID NO:16) and RHWPGGFDY (SEQ ID NO:3), respectively, or   (b) the light chain or antigen binding fragment thereof further comprises an HVR-L1, HVR-L2 and an HVR-L3 sequence having at least 85% sequence identity to RASQDVSTAVA (SEQ ID NO:17), SASFLYS (SEQ ID NO:18) and QQYLYHPAT (SEQ ID NO:19), respectively.   
     
     
         26 . The nucleic acid of  claim 25 , wherein sequence identity is 90%. 
     
     
         27 . The nucleic acid of  claim 26 , wherein the sequence identity is 95%. 
     
     
         28 . The nucleic acid of  claim 25 , wherein the anti-PD-L1 antibody further comprises a VL and a VH framework region derived from a human consensus sequence. 
     
     
         29 . The nucleic acid of  claim 28 , wherein the VH sequence is derived from a Kabat subgroup I, II, or III sequence. 
     
     
         30 . The nucleic acid of  claim 29 , wherein the VH sequence is derived from Kabat subgroup III. 
     
     
         31 . The nucleic acid of  claim 30 , further wherein the heavy chain framework sequences are juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4). 
     
     
         32 . The nucleic acid of  claim 31 , wherein one or more of the heavy chain framework sequences is the following: 
       
         
           
                 
                 
               
                     
                   HC-FR1 is 
                 
                 
               
                   (SEQ ID NO: 4) 
                 
                 
                 
               
                     
                   EVQLVESGGGLVQPGGSLRLSCAAS; 
                 
                     
                     
                 
                     
                   HC-FR2 is 
                 
                 
               
                   (SEQ ID NO: 5) 
                 
                 
                 
               
                     
                   WVRQAPGKGLEWV; 
                 
                     
                     
                 
                     
                   HC-FR3 is 
                 
                 
               
                   (SEQ ID NO: 6) 
                 
                 
                 
               
                     
                   RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR; 
                 
                     
                     
                 
                     
                   HC-FR4 is 
                 
                 
               
                   (SEQ ID NO: 7) 
                 
                 
                 
               
                     
                   WGQGTLVTVSA. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         33 . The nucleic acid of  claim 28 , wherein the VL sequence is derived from a Kabat kappa I, II, III or IV subgroup sequence. 
     
     
         34 . The nucleic acid of  claim 33 , wherein the VL sequence is derived from Kabat kappa I. 
     
     
         35 . The nucleic acid of  claim 33 , further wherein the encoded variable light chain framework sequences are juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). 
     
     
         36 . The nucleic acid of  claim 35 , wherein one or more of the light chain framework sequences is the following: 
       
         
           
                 
                 
               
                     
                   LC-FR1 is 
                 
                 
               
                   (SEQ ID NO: 11) 
                 
                 
                 
               
                     
                   DIQMTQSPSSLSASVGDRVTITC; 
                 
                     
                     
                 
                     
                   LC-FR2 is 
                 
                 
               
                   (SEQ ID NO: 12) 
                 
                 
                 
               
                     
                   WYQQKPGKAPKLLIY; 
                 
                     
                     
                 
                     
                   LC-FR3 is 
                 
                 
               
                   (SEQ ID NO: 13) 
                 
                 
                 
               
                     
                   GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC,; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   LC-FR4 is 
                 
                 
               
                   (SEQ ID NO: 14) 
                 
                 
                 
               
                     
                   FGQGTKVEIKR. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         37 . The nucleic acid of  claim 25 , wherein the anti-PD-L1 antibody comprises a constant region derived from a murine antibody. 
     
     
         38 . The nucleic acid of  claim 25 , wherein the anti-PD-L1 antibody comprises a constant region derived from a human antibody. 
     
     
         39 . The nucleic acid of  claim 37 , wherein the constant region is IgG2A. 
     
     
         40 . The nucleic acid of  claim 38 , wherein the constant region is IgG1. 
     
     
         41 . The nucleic acid of  claim 40 , wherein the encoded antibody has reduced or minimal effector function. 
     
     
         42 . The nucleic acid of  claim 41 , wherein the minimal effector function results from an effector-less Fc mutation. 
     
     
         43 . The nucleic acid of  claim 42 , wherein the effector-less Fc mutation is N297A. 
     
     
         44 . The nucleic acid of  claim 42 , wherein the effector-less Fc mutation is D265A/N297A. 
     
     
         45 . An isolated nucleic acid encoding an anti-PD-L1 antibody or antigen binding fragment thereof, wherein the antibody or antibody fragment comprises a heavy chain and a light chain variable region sequence, wherein:
 (a) the heavy chain comprises an HVR-H1, HVR-H2 and HVR-H3, wherein further:
 (i) the HVR-H1 sequence is GFTFSX 1 SWIH (SEQ ID NO:1); 
 (ii) the HVR-H2 sequence is AWIX 2 PYGGSX 3 YYADSVKG (SEQ ID NO:2); 
 (iii) the HVR-H3 sequence is RHWPGGFDY, and (SEQ ID NO:3); 
   (b) the light chain comprises an HVR-L1, HVR-L2 and HVR-L3, wherein further:
 (iv) the HVR-L1 sequence is RASQX 4 X 5 X 6 TX 7 X 8 A (SEQ ID NOs:8); 
 (v) the HVR-L2 sequence is SASX 9 LX 10 S (SEQ ID NOs:9); 
 (vi) the HVR-L3 sequence is QQX 11 X 12 X 13 X 14 PX 15 T (SEQ ID NOs:10); 
   wherein: X 1  is D or G; X 2  is S or L; X 3  is T or S; X 4  may be D or V; X 5  may be V or I; X 6  may be S or N; X 7  may be A or F; X 8  may be V or L; X 9  may be F or T; X 10  may be Y or A; X 11  may be Y, G, F, or S; X 12  may be L, Y, F or W; X 13  may be Y, N, A, T, G, F or I; X 14  may be H, V, P, T or I; X 15  may be A, W, R, P or T.   
     
     
         46 . The nucleic acid of  claim 45  wherein X 1  is D; X 2  is S and X 3  is T. 
     
     
         47 . The nucleic acid of  claim 45 , wherein X 4 =D, X 5 =V, X 6 =5, X 7 =A and X 8 =V, X 9 =F, and X 10 =Y, X 11 =Y, X 12 =L, X 13 =Y, X 14 =H and X 15 =A. 
     
     
         48 . The nucleic acid of  claim 45 , wherein X 1 =D, X 2 =S and X 3 =T, X 4 =D, X 5 =V, X 6 =S, X 7 =A and X 8 =V, X 9 =F, and X 10 =Y, X 11 =Y, X 12 =L, X 13 =Y, X 14 =H and X 15 =A. 
     
     
         49 . The nucleic acid of  claim 46 , wherein the encoded variable region heavy chain framework sequences are juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4). 
     
     
         50 . The nucleic acid of  claim 49 , wherein the framework sequences are human. 
     
     
         51 . The nucleic acid of  claim 50  wherein the framework sequences are VH subgroup III consensus framework. 
     
     
         52 . The nucleic acid of  claim 51  wherein one or more of the framework sequences is the following: 
       
         
           
                 
                 
               
                     
                   HC-FR1 is 
                 
                 
               
                   (SEQ ID NO: 4) 
                 
                 
                 
               
                     
                   EVQLVESGGGLVQPGGSLRLSCAAS; 
                 
                     
                     
                 
                     
                   HC-FR2 is 
                 
                 
               
                   (SEQ ID NO: 5) 
                 
                 
                 
               
                     
                   WVRQAPGKGLEWV; 
                 
                     
                     
                 
                     
                   HC-FR3 is 
                 
                 
               
                   (SEQ ID NO: 6) 
                 
                 
                 
               
                     
                   RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR; 
                 
                     
                     
                 
                     
                   HC-FR4 is 
                 
                 
               
                   (SEQ ID NO: 7) 
                 
                 
                 
               
                     
                   WGQGTLVTVSA. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         53 . The nucleic acid of  47 , wherein the encoded variable region light chain framework sequences juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). 
     
     
         54 . The nucleic acid of  claim 53  wherein the variable region light chain framework sequences are VL kappa I consensus framework. 
     
     
         55 . The nucleic acid of  claim 54  wherein one or more of the framework sequences is the following: 
       
         
           
                 
                 
               
                     
                   LC-FR1 is 
                 
                 
               
                   (SEQ ID NO: 11) 
                 
                 
                 
               
                     
                   DIQMTQSPSSLSASVGDRVTITC; 
                 
                     
                     
                 
                     
                   LC-FR2 is 
                 
                 
               
                   (SEQ ID NO: 12) 
                 
                 
                 
               
                     
                   WYQQKPGKAPKLLIY; 
                 
                     
                     
                 
                     
                   LC-FR3 is 
                 
                 
               
                   (SEQ ID NO: 13) 
                 
                 
                 
               
                     
                   GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC,; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   LC-FR4 is 
                 
                 
               
                   (SEQ ID NO: 14) 
                 
                 
                 
               
                     
                   FGQGTKVEIKR. 
                 
             
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
               
            
             
                
               
            
             
                
                
                
                
               
            
             
                
               
            
             
                
               
            
           
         
       
     
     
         56 . The nucleic acid of  claim 48  wherein the antibody or antibody fragment comprises:
 (a) variable region heavy chain framework sequences juxtaposed between the HVRs according to the formula: (HC-FR1)-(HVR-H1)-(HC-FR2)-(HVR-H2)-(HC-FR3)-(HVR-H3)-(HC-FR4), and 
 (b) variable region light chain framework sequences juxtaposed between the HVRs according to the formula: (LC-FR1)-(HVR-L1)-(LC-FR2)-(HVR-L2)-(LC-FR3)-(HVR-L3)-(LC-FR4). 
 
       wherein the variable heavy chain framework sequences are the following:
 (i) HC-FR1 is EVQLVESGGGLVQPGGSLRLSCAAS (SEQ ID NO:4); 
 (ii) HC-FR2 is WVRQAPGKGLEWV (SEQ ID NO:5); 
 (iii) HC-FR3 is RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (SEQ ID NO:6); 
 (iv) HC-FR4 is WGQGTLVTVSA; and (SEQ ID NO:7); and 
 
       further wherein the variable light chain framework sequences are the following:
 (i) LC-FR1 is DIQMTQSPSSLSASVGDRVTITC (SEQ ID NO:11); 
 (ii) LC-FR2 is WYQQKPGKAPKLLIY (SEQ ID NO:12); 
 (iii) LC-FR3 is GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (SEQ ID NO:13); 
 (iv) LC-FR4 is FGQGTKVEIKR (SEQ ID NO:14). 
 
     
     
         57 . The nucleic acid of  claim 56  further comprising a human constant region. 
     
     
         58 . The nucleic acid of  claim 57 , wherein the constant region is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4. 
     
     
         59 . The nucleic acid of  claim 58  wherein the constant region is IgG1. 
     
     
         60 . The nucleic acid of  claim 59  having reduced or minimal effector function. 
     
     
         61 . The nucleic acid of  claim 60 , wherein the minimal effector function results from an effector-less Fc mutation. 
     
     
         62 . The nucleic acid of  claim 61 , wherein the effector-less Fc mutation is N297A. 
     
     
         63 . The nucleic acid of  claim 61 , wherein the effector-less Fc mutation is D265A/N297A. 
     
     
         64 . The nucleic acid of  claim 60 , wherein the minimal effector function results from aglycosylation. 
     
     
         65 . The nucleic acid of  claim 56 , further comprising a murine constant region. 
     
     
         66 . The nucleic acid of  claim 65 , wherein the constant region is selected from the group consisting of IgG1, IgG2A, IgG2B and IgG3. 
     
     
         67 . The nucleic acid of  claim 66 , wherein the constant region is IgG2A. 
     
     
         68 . The nucleic acid of  claim 67 , which has reduced or minimal effector function. 
     
     
         69 . The nucleic acid of  claim 68 , wherein the minimal effector function results from an effector-less Fc mutation. 
     
     
         70 . The nucleic acid of  claim 69 , wherein the effector-less Fc mutation is N297A. 
     
     
         71 . The nucleic acid of  claim 69 , wherein the effector-less Fc mutation is D265A/N297A. 
     
     
         72 . The nucleic acid of  claim 68 , wherein the minimal effector function results from aglycosylation. 
     
     
         73 . An isolated nucleic acid encoding anti-PD-L1 antibody or antigen binding fragment thereof, wherein the antibody or antibody fragment comprises a heavy chain and light chain variable region sequence, wherein:
 (a) the heavy chain comprises the sequence: EVQLVESGGGLVQPGGSLRLS CAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTI SADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVS A (SEQ ID NO:20), and   (b) the light chain comprises the sequence: DIQMTQSPSSLSASVGDRVTITC RASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTL TISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR (SEQ ID NO:21).   
     
     
         74 . The nucleic acid of  claim 73  further comprising a human constant region. 
     
     
         75 . The nucleic acid of  claim 74 , wherein the constant region is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4. 
     
     
         76 . The nucleic acid of  claim 75 , wherein the constant region is IgG1. 
     
     
         77 . The nucleic acid of  claim 76  having reduced or minimal effector function. 
     
     
         78 . The nucleic acid of  claim 77 , wherein the minimal effector function results from an effector-less Fc mutation. 
     
     
         79 . The nucleic acid of  claim 78 , wherein the effector-less Fc mutation is N297A. 
     
     
         80 . The nucleic acid of  claim 78 , wherein the effector-less Fc mutation is D265A/N297A. 
     
     
         81 . The nucleic acid of  claim 77 , wherein the minimal effector function results from aglycosylation. 
     
     
         82 . A vector comprising the nucleic acid of any of  claims 1 - 12 . 
     
     
         83 . (canceled) 
     
     
         84 . A vector comprising the nucleic acid of any of  claims 25 - 44 . 
     
     
         85 . A vector comprising the nucleic acid of any of  claims 45 - 72 . 
     
     
         86 . A vector comprising the nucleic acid of any of  claims 73 - 81 . 
     
     
         87 . A host cell comprising the vector of  claim 82 . 
     
     
         88 . A host cell comprising the vector of  claim 84 . 
     
     
         89 . A host cell comprising the vector of  claim 85 . 
     
     
         90 . A host cell comprising the vector of  claim 86 . 
     
     
         91 . The host cell of any of  claims 87 - 90 , which is eukaryotic. 
     
     
         92 . The host cell of  claim 91 , which is mammalian. 
     
     
         93 . The host cell of  claim 92 , which is a Chinese Hamster Ovary (CHO) cells. 
     
     
         94 . The host cell of any of  claims 87 - 90 , which is prokaryotic. 
     
     
         95 . The host cell of  claim 94 , which is  E. coli.    
     
     
         96 . A process for making an anti-PD-L1 antibody comprising culturing the host cell of  claim 93  under conditions suitable for the expression of the vector encoding the anti-PD-L1 antibody or antigen binding fragment, and recovering the antibody or fragment.

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