Anti-pd-1 antibodies, activatable anti-pd-1 antibodies, and methods of use thereof

Abstract

The invention relates generally to antibodies that specifically bind programmed cell death protein 1 (PD-1), activatable antibodies that specifically bind to PD-1 and methods of making and using these anti-PD-1 antibodies and anti-PD-1 activatable antibodies in a variety of therapeutic, diagnostic and prophylactic indications.

1 - 99 . (canceled) 
     
     
         100 . An isolated antibody or antigen binding fragment thereof (AB) that specifically binds to a human PD-1, wherein the AB comprises:
 a VH CDR1 amino acid sequence comprising GFTFSNYAMS SEQ ID NO: 655),   a VH CDR2 amino acid sequence comprising YISNGGGDTH (SEQ ID NO: 660),   a VH CDR3 amino acid sequence comprising ENYGTSPFVY (SEQ ID NO: 666),   a VL CDR1 amino acid sequence comprising RASESVDNYGIS (SEQ ID NO: 673),   a VL CDR2 amino acid sequence comprising AASNQGS (SEQ ID NO: 678), and   a VL CDR3 amino acid sequence comprising QQSKDVPWT (SEQ ID NO: 683).   
     
     
         101 . An activatable antibody that, when activated, specifically binds to a human PD-1, wherein said activatable antibody comprises:
 an antibody or an antigen binding fragment thereof (AB) that specifically binds to mammalian PD-1, wherein the AB comprises   a VH CDR1 amino acid sequence comprising GFTFSNYAMS SEQ ID NO: 655),   a VH CDR2 amino acid sequence comprising YISNGGGDTH (SEQ ID NO: 660),   a VH CDR3 amino acid sequence comprising ENYGTSPFVY (SEQ ID NO: 666),   a VL CDR1 amino acid sequence comprising RASESVDNYGIS (SEQ ID NO: 673),   a VL CDR2 amino acid sequence comprising AASNQGS (SEQ ID NO: 678), and   a VL CDR3 amino acid sequence comprising QQSKDVPWT (SEQ ID NO: 683);   a masking moiety (MM) that inhibits the binding of the AB to mammalian PD-1 when the activatable antibody is in an uncleaved state; and   a cleavable moiety (CM) coupled to the AB, wherein the CM is a polypeptide that functions as a substrate for a protease,   wherein the MM is coupled to the AB via the CM.   
     
     
         102 . The activatable antibody of  claim 101 , wherein the AB specifically blocks a human PD-L1 and a human PD-L2 from binding to the human PD-1. 
     
     
         103 . The activatable antibody of  claim 101 , wherein the activatable antibody in an uncleaved state specifically binds to the human PD-1 with a dissociation constant of 0.5 nM to 1 nM. 
     
     
         104 . The activatable antibody of  claim 101 , wherein the CM is a substrate for a protease that is active in diseased tissue. 
     
     
         105 . A pharmaceutical composition comprising the activatable antibody of  claim 101  and a carrier. 
     
     
         106 . An isolated nucleic acid molecule encoding the activatable antibody of  claim 101 . 
     
     
         107 . A vector comprising the isolated nucleic acid molecule of  claim 106 . 
     
     
         108 . A method of producing an activatable antibody by culturing a cell under conditions that lead to expression of the activatable antibody, wherein the cell comprises the nucleic acid molecule of  claim 106 . 
     
     
         109 . A method of reducing binding of a ligand selected from the group consisting of PD-L1 or PD-L2 to PD-1 on T cells comprising administering an effective amount of the activatable antibody of  claim 101  to a subject in need thereof. 
     
     
         110 . A method of reducing immune suppression mediated by engagement of PD-1 on T cells to PD-L1 or PD-L2 on tumor or other immune cells, the method comprising administering to a subject in need thereof a therapeutically effective amount of the activatable antibody of  claim 101 . 
     
     
         111 . A method of treating, alleviating a symptom of, or delaying the progression of a cancer comprising administering to a subject in need thereof a therapeutically effective amount of the activatable antibody of  claim 101 . 
     
     
         112 . The activatable antibody of  claim 101 , wherein the activatable antibody comprises a heavy chain that comprises an IgG4 amino acid sequence. 
     
     
         113 . The activatable antibody of  claim 101 , wherein the AB comprises a VH comprising the amino acid sequence of SEQ ID NO: 33. 
     
     
         114 . The activatable antibody of  claim 101 , wherein the AB comprises a VL comprising the amino acid sequence of SEQ ID NO: 41. 
     
     
         115 . The activatable antibody of  claim 101 , wherein the MM comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 66-213, 384-514, and 548-571. 
     
     
         116 . The activatable antibody of  claim 101 , wherein the CM comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 214, 294-361, 1092-1112, 1157, 1158, 1161, 1162, 1165, 1166, 1169, 1520, and 1695-1704. 
     
     
         117 . The activatable antibody of  claim 101 , wherein the AB has one or more of the characteristics selected from the group consisting of:
 (a) the AB is an antigen binding fragment selected from the group consisting of a Fab fragment, a F(ab′)2 fragment, a scFv, a scAb, a dAb, a single domain heavy chain antibody, and a single domain light chain antibody;   (b) the AB is a monoclonal antibody;   (c) the AB is linked to the CM;   (d) the AB is linked directly to the CM; and   (e) the AB is linked to the CM via a linking peptide.   
     
     
         118 . A conjugated activatable antibody comprising the activatable antibody of  claim 101  conjugated to an agent. 
     
     
         119 . The conjugated activatable antibody of  claim 118 , wherein the agent has one or more of the following characteristics selected from the group consisting of:
 (a) the agent is conjugated to the activatable antibody via a linker;   (b) the agent is conjugated to the activatable antibody via a cleavable linker,   (c) the linker is conjugated to the activatable antibody via a non-cleavable linker;   (d) the agent is a detectable moiety; and   (e) the agent is a diagnostic agent.