FIELD OF THE INVENTION

Antibodies that bind to Programmed Death-1 protein (PD-1) are provided. Methods of treatment comprising administering anti-PD-1 antibodies are also provided.

BACKGROUND

The Programmed Death 1 (PD-1) protein is an inhibitory member of the CD28 family of receptors, which also includes CD28, CTLA-4, ICOS and BTLA. PD-1 is expressed on the surface of activated B cells, T cells, and myeloid cells. PD-1 contains a membrane proximal immunoreceptor tyrosine inhibitory motif (ITIM) and a membrane distal tyrosine-based switch motif (ITSM). Although structurally similar to CTLA-4, PD-1 lacks the MYPPY motif that is critical for B7-1 and B7-2 binding. In addition, although CD28, ICOS and CTLA-4 (other members of the CD28 family) all have an unpaired cysteine residue allowing for homodimerization, PD-1 is believed to exist as a monomer, lacking the unpaired cysteine residue characteristic in other CD28 family members. The PD-1 receptor has two ligands, PD-ligand-1 (PD-L1) and PD-L2. The term “PD-L1” refers to the ligand of the PD-1 receptor also known as CD274 and B7H 1. PD-L1 is a 290 amino acid protein with an extracellular IgV-like domain, an extracellular IgC-like domain, a transmembrane domain and a highly conserved intracellular domain of approximately 30 amino acids. PD-L1 is constitutively expressed on many cells such as antigen presenting cells (e.g., dendritic cells, macrophages, and B-cells) and on hematopoietic and non-hematopoietic cells (e.g., vascular endothelial cells, pancreatic islets, and sites of immune privilege). The term “PD-L2” refers to the ligand of the PD-1 receptor also known as CD273 and B7-DC. PD-L2 has an extracellular IgV-like domain, an extracellular IgC-like domain, a transmembrane domain and an intracellular domain of approximately 30 amino acids in humans. PD-L2 has a more restricted expression than PD-L1, with its expression largely confined to hematopoietic cells including macrophages, dendritic cells, some B cell subsets and bone marrow-derived mast cells.

PD-1 functions as an immune checkpoint and works to prevent the activation of T-cells. PD-1 antagonists activate the immune system to attack tumors and have shown success in treating cancers, and in some instances, with less toxicity than other chemotherapeutic treatments. PD-1 antagonists can also be used in combination regimens with other chemotherapeutic agents. Currently approved PD-1 antagonists include anti-PD-1 antibodies, Opdivo® and Keytruda®, and anti-PD-L1 antibody, Tecentriq™.

There remains a need for additional antagonists of PD-1 for treatment of cancer and other diseases and disorders.

SUMMARY

Antibodies that bind Programmed Death 1 (PD-1) are provided, wherein the antibodies bind human PD-1 and mouse PD-1, and wherein the antibodies block binding of PD-L1 and/or PD-L2 to PD-1.

In some embodiments, an isolated antibody that binds to Programmed Death 1 (PD-1) is provided, wherein the antibody binds to an epitope comprising amino acids 126 to 136 of human PD-1. In some embodiments, amino acids 126 to 136 are numbered according to SEQ ID NO: 1. In some embodiments, the antibody binds to human PD-1 of SEQ ID NO: 382 with at least 10-fold greater affinity than the antibody binds to one or more PD-1 variants selected from I126A, L128A, A132L, I134A, and E136A.

In some embodiments, the antibody:a) binds to human PD-1 ECD-Fc of SEQ ID NO: 401 with at least 10-fold greater affinity than the antibody binds to PD-1 variant I126A ECD-Fc (SEQ ID NO: 389); and/orb) binds to human PD-1 ECD-Fc of SEQ ID NO: 401 with at least 10-fold greater affinity than the antibody binds to PD-1 variant L128A ECD-Fc (SEQ ID NO: 390); and/orc) binds to human PD-1 ECD-Fc of SEQ ID NO: 401 with at least 10-fold greater affinity than the antibody binds to PD-1 variant A132L ECD-Fc (SEQ ID NO: 391); and/ord) binds to human PD-1 ECD-Fc of SEQ ID NO: 401 with at least 10-fold greater affinity than the antibody binds to PD-1 variant I134A ECD-Fc (SEQ ID NO: 392); and/ore) binds to human PD-1 ECD-Fc of SEQ ID NO: 401 with at least 10-fold greater affinity than the antibody binds to PD-1 variant E136A ECD-Fc (SEQ ID NO: 393).

In some embodiments, the antibody binds to mouse PD-1 ECD-Fc of SEQ ID NO: 403. In some embodiments, the antibody binds to mouse PD-1 ECD-Fc of SEQ ID NO: 403 with at least 10-fold greater affinity than the antibody binds to mouse PD-1 variant H129P ECD-Fc (SEQ I NO: 394). In some embodiments, the antibody binds to rat PD-1 variant P129H ECD-Fc (SEQ ID NO: 397) with at least 10-fold greater affinity than the antibody binds to rat PD-1 ECD-Fc (SEQ ID NO: 405).

In various embodiments, affinity is determined by biolayer interferometry.

In some embodiments, the antibody comprises (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27. In some embodiments, the antibody comprises a heavy chain variable region (VH) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 20 and a light chain variable region (VL) that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibody comprises a VHcomprising the amino acid sequence of SEQ ID NO: 20 and a VLcomprising the amino acid sequence of SEQ ID NO: 24.

In some embodiments, an antibody that binds to Programmed Death 1 (PD-1) is provided, wherein the antibody comprises:i) (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 5; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 6; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 7; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 9; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 10; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 11; orii) (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 13; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 14; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 15; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 17; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 18; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 19; oriii) (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 21; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 22; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 23; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 25; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 26; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 27; oriv) (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 29; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 30; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 31; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 33; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 34; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 35; orv) (a) HCDR1 comprising the amino acid sequence of SEQ ID NO: 37; (b) HCDR2 comprising the amino acid sequence of SEQ ID NO: 38; (c) HCDR3 comprising the amino acid sequence of SEQ ID NO: 39; (d) LCDR1 comprising the amino acid sequence of SEQ ID NO: 41; (e) LCDR2 comprising the amino acid sequence of SEQ ID NO: 42; and (f) LCDR3 comprising the amino acid sequence of SEQ ID NO: 43; orvi) (a) HCDR1 comprising an amino acid sequence selected from SEQ ID NOs: 45, 53, 61, 69, 77, 85, 93, 101, 109 and 117; (b) HCDR2 comprising an amino acid sequence selected from SEQ ID NOs: 46, 54, 62, 70, 78, 86, 94, 102, 110 and 118; (c) HCDR3 comprising an amino acid sequence selected from SEQ ID NOs: 47, 55, 63, 71, 79, 87, 95, 103, 111, and 119; (d) LCDR1 comprising an amino acid sequence selected from SEQ ID NOs: 49, 57, 65, 73, 81, 89, 97, 105, 113, and 121; (e) LCDR2 comprising an amino acid sequence selected from SEQ ID NOs: 50, 58, 66, 74, 82, 90, 98, 106, 114, and 122; and (f) LCDR3 comprising an amino acid sequence selected from SEQ ID NOs: 51, 59, 67, 75, 83, 91, 99, 107, 115, and 123; orvii)(a) HCDR1 comprising an amino acid sequence selected from SEQ ID NOs: 125, 133, 141, 149, 157, 165, 173, 181, 189, 197, 205, 213, 221, 229, 237, 245, 253, 261, 269, 277 and 285; (b) HCDR2 comprising an amino acid sequence selected from SEQ ID NOs: 126, 134, 142, 150, 158, 166, 174, 182, 190, 198, 206, 214, 222, 230, 238, 246, 254, 262, 270, 278 and 286; (c) HCDR3 comprising an amino acid sequence selected from SEQ ID NOs: 127, 135, 143, 151, 159, 167, 175, 183, 191, 199, 207, 215, 223, 231, 239, 247, 255, 263, 271, 279 and 287; (d) LCDR1 comprising an amino acid sequence selected from SEQ ID NOs: 129, 137, 145, 153, 161, 169, 177, 185, 193, 201, 209, 217, 225, 233, 241, 249, 257, 265, 273, 281 and 289; (e) LCDR2 comprising an amino acid sequence selected from SEQ ID NOs: 130, 138, 146, 154, 162, 170, 178, 186, 194, 202, 210, 218, 226, 234, 242, 250, 258, 266, 274, 282, and 290; and (f) LCDR3 comprising an amino acid sequence selected from SEQ ID NOs: 131, 139, 147, 155, 163, 171, 179, 187, 195, 203, 211, 219, 227, 235, 243, 251, 259, 267, 275, 283 and 291; orviii) (a) HCDR1 comprising an amino acid sequence selected from SEQ ID NOs: 293, 301, 309 and 317; (b) HCDR2 comprising an amino acid sequence selected from SEQ ID NOs: 294, 302, 310 and 318; (c) HCDR3 comprising an amino acid sequence selected from SEQ ID NOs: 295, 303, 311 and 319; (d) LCDR1 comprising an amino acid sequence selected from SEQ ID NOs: 297, 305, 313, and 321; (e) LCDR2 comprising an amino acid sequence selected from SEQ ID NOs: 298, 306, 314 and 322; and (f) LCDR3 comprising an amino acid sequence selected from SEQ ID NOs: 299, 307, 315, and 323.

In some embodiments, an antibody that binds PD-1 is provided, which comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:i) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 4 and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 8; orii) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 12 and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 16; oriii) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 20 and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 24; oriv) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 28 and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 32; orv) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 36 and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 40; orvi) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 44, 52, 60, 68, 76, 84, 92, 100, 108 and 116; and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 48, 56, 64, 72, 80, 88, 96, 104, 112, and 120; orvii) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 124, 132, 140, 148, 156, 164, 172, 180, 188, 196, 204, 212, 220, 228, 236, 244, 252, 260, 268, 276, and 284; and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 128, 136, 144, 152, 160, 168, 176, 184, 192, 200, 208, 216, 224, 232, 240, 248, 256, 264, 272, 280 and 288; orviii) the VHis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 292, 300, 308 and 316; and the VLis at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs: 296, 304, 312, and 320.

In some embodiments, an antibody that binds PD-1 is provided, which comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein:i) the VHcomprises the amino acid sequence of SEQ ID NO: 4 and the VLcomprises the amino acid sequence of SEQ ID NO: 8; orii) the VHcomprises the amino acid sequence of SEQ ID NO: 12 and the VLcomprises the amino acid sequence of SEQ ID NO: 16; oriii) the VHcomprises the amino acid sequence of SEQ ID NO: 20 and the VLcomprises the amino acid sequence of SEQ ID NO: 24; oriv) the VHcomprises the amino acid sequence of SEQ ID NO: 28 and the VLcomprises the amino acid sequence of SEQ ID NO: 32; orv) the VHcomprises the amino acid sequence of SEQ ID NO: 36 and the VLcomprises the amino acid sequence of SEQ ID NO: 40; orvi) the VHcomprises the amino acid sequence of SEQ ID NO: 44 and the VLcomprises the amino acid sequence of SEQ ID NO: 48; orvii) the VHcomprises the amino acid sequence of SEQ ID NO: 52 and the VLcomprises the amino acid sequence of SEQ ID NO: 56; orviii) the VHcomprises the amino acid sequence of SEQ ID NO: 60 and the VLcomprises the amino acid sequence of SEQ ID NO: 64; orix) the VHcomprises the amino acid sequence of SEQ ID NO: 68 and the VLcomprises the amino acid sequence of SEQ ID NO: 72; orx) the VHcomprises the amino acid sequence of SEQ ID NO: 76 and the VLcomprises the amino acid sequence of SEQ ID NO: 80; orxi) the VHcomprises the amino acid sequence of SEQ ID NO: 84 and the VLcomprises the amino acid sequence of SEQ ID NO: 88; orxii) the VHcomprises the amino acid sequence of SEQ ID NO: 92 and the VLcomprises the amino acid sequence of SEQ ID NO: 96; orxiii) the VHcomprises the amino acid sequence of SEQ ID NO: 100 and the VLcomprises the amino acid sequence of SEQ ID NO: 104; orxiv) the VHcomprises the amino acid sequence of SEQ ID NO: 108 and the VLcomprises the amino acid sequence of SEQ ID NO: 112; orxv) the VHcomprises the amino acid sequence of SEQ ID NO: 116 and the VLcomprises the amino acid sequence of SEQ ID NO: 120; orxvi) the VHcomprises the amino acid sequence of SEQ ID NO: 124 and the VLcomprises the amino acid sequence of SEQ ID NO: 128; orxvii) the VHcomprises the amino acid sequence of SEQ ID NO: 132 and the VLcomprises the amino acid sequence of SEQ ID NO: 136; orxviii) the VHcomprises the amino acid sequence of SEQ ID NO: 140 and the VLcomprises the amino acid sequence of SEQ ID NO: 144; orxix) the VHcomprises the amino acid sequence of SEQ ID NO: 148 and the VLcomprises the amino acid sequence of SEQ ID NO: 152; orxx) the VHcomprises the amino acid sequence of SEQ ID NO: 156 and the VLcomprises the amino acid sequence of SEQ ID NO: 160; orxxi) the VHcomprises the amino acid sequence of SEQ ID NO: 164 and the VLcomprises the amino acid sequence of SEQ ID NO: 168; orxxii) the VHcomprises the amino acid sequence of SEQ ID NO: 172 and the VLcomprises the amino acid sequence of SEQ ID NO: 176; orxxiii) the VHcomprises the amino acid sequence of SEQ ID NO: 180 and the VLcomprises the amino acid sequence of SEQ ID NO: 184; orxxiv) the VHcomprises the amino acid sequence of SEQ ID NO: 188 and the VLcomprises the amino acid sequence of SEQ ID NO: 192; orxxv) the VHcomprises the amino acid sequence of SEQ ID NO: 196 and the VLcomprises the amino acid sequence of SEQ ID NO: 200; orxxvi) the VHcomprises the amino acid sequence of SEQ ID NO: 204 and the VLcomprises the amino acid sequence of SEQ ID NO: 208; orxxvii) the VHcomprises the amino acid sequence of SEQ ID NO: 212 and the VLcomprises the amino acid sequence of SEQ ID NO: 216; orxxviii) the VHcomprises the amino acid sequence of SEQ ID NO: 220 and the VLcomprises the amino acid sequence of SEQ ID NO: 224; orxxix) the VHcomprises the amino acid sequence of SEQ ID NO: 228 and the VLcomprises the amino acid sequence of SEQ ID NO: 232; orxxx) the VHcomprises the amino acid sequence of SEQ ID NO: 236 and the VLcomprises the amino acid sequence of SEQ ID NO: 240; orxxxi) the VHcomprises the amino acid sequence of SEQ ID NO: 244 and the VLcomprises the amino acid sequence of SEQ ID NO: 248; orxxxii) the VHcomprises the amino acid sequence of SEQ ID NO: 252 and the VLcomprises the amino acid sequence of SEQ ID NO: 256; orxxxiii) the VHcomprises the amino acid sequence of SEQ ID NO: 260 and the VLcomprises the amino acid sequence of SEQ ID NO: 264; orxxxiv) the VHcomprises the amino acid sequence of SEQ ID NO: 268 and the VLcomprises the amino acid sequence of SEQ ID NO: 272; orxxxv) the VHcomprises the amino acid sequence of SEQ ID NO: 276 and the VLcomprises the amino acid sequence of SEQ ID NO: 280; orxxxvi) the VHcomprises the amino acid sequence of SEQ ID NO: 284 and the VLcomprises the amino acid sequence of SEQ ID NO: 288; orxxxvii) the VHcomprises the amino acid sequence of SEQ ID NO: 292 and the VLcomprises the amino acid sequence of SEQ ID NO: 296; orxxxviii) the VHcomprises the amino acid sequence of SEQ ID NO: 300 and the VLcomprises the amino acid sequence of SEQ ID NO: 304; orxxxix) the VHcomprises the amino acid sequence of SEQ ID NO: 308 and the VLcomprises the amino acid sequence of SEQ ID NO: 312; orxl) the VHcomprises the amino acid sequence of SEQ ID NO: 316 and the VLcomprises the amino acid sequence of SEQ ID NO: 320.

In some embodiments, an antibody that competes with an antibody provided herein for binding to human PD-1 is provided, wherein the antibody binds to human PD-1 and mouse PD-1, and where in the antibody inhibits binding of human PD-1 to human PD-L1, inhibits binding of human PD-1 to human PD-L2, inhibits binding of mouse PD-1 to mouse PD-L1, and inhibits binding of mouse PD-1 to mouse PD-L2.

In some embodiments, an antibody provided herein binds to human PD-1 with an affinity (KD) of less than 5 nM. In some embodiments, an antibody provided herein binds to mouse PD-1 with an affinity (KD) of less than 10 nM. In some embodiments, affinity is determined using biolayer interferometry.

In some embodiments, an antibody provided herein is a monoclonal antibody. In some embodiments, an antibody provided herein is a human antibody, chimeric antibody, or a humanized antibody. In some embodiments, an antibody provided herein is an antibody fragment selected from a Fab, Fab′, Fv, scFv or (Fab′)2fragment. In some embodiments, an antibody provided herein is a full length antibody. In some embodiments, an antibody provided herein is an IgG1 or IgG4 antibody.

In some embodiments, an antibody provided herein binds human PD-1. In some embodiments, human PD-1 comprises the amino acid sequence of SEQ ID NO: 382. In some embodiments, an antibody provided herein binds mouse PD-1. In some embodiments, mouse PD-1 comprises the amino acid sequence of SEQ ID NO: 383. In some embodiments, an antibody provided herein binds cynomolgus monkey PD-1. In some embodiments, cynomolgus monkey PD-1 comprises the amino acid sequence of SEQ ID NO: 384.

In some embodiments, an antibody provided herein inhibits binding of PD-1 to PD-L1. In some embodiments, an antibody provided herein inhibits binding of PD-1 to PD-L2. In some embodiments, an antibody provided herein inhibits binding of PD-1 to PD-L1 and inhibits the binding of PD-1 to PD-L2.

In some embodiments, administration of an antibody provided herein to a mammal increases the level of at least one cytokine selected from IFNγ and IL-2. In some embodiments, an antibody provided herein increases the level of at least one cytokine selected from IFNγ and IL-2 by at least 2-fold. In some embodiments, the level of the cytokine is measured 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 36 hours, or 48 hours after administration of the antibody. In some embodiments, at least one cytokine is IFNγ. In some embodiments, at least one cytokine is IL-2. In some embodiments, the level of the chemokine is measured 24 hours after administration of the antibody.

In some embodiments, administration of an antibody provided herein to a mammal enhances an immune response in the mammal. In some embodiments, administration of an antibody provided herein to a mammal results in activation of T cells in the mammal. In some embodiments, administration of an antibody provided herein to a mammal reduces tumor size in a mammal with cancer. In some embodiments, the mammal is a human. In some embodiments, the human has cancer. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

In some embodiments, an isolated nucleic acid is provided, which encodes an antibody provided herein. In some embodiments, a vector is provided, which comprises the nucleic acid. In some embodiments, a host cell is provided, which comprises the vector. In some embodiments, a host cell is provided, which produces an antibody provided herein. In some embodiments, a method for making an anti-PD-1 antibody is provided, comprising culturing the host cell under conditions suitable for expression of the antibody. In some embodiments, the method further comprises recovering the antibody produced by the host cell.

In some embodiments, a pharmaceutical composition is provided, comprising an anti-PD-1 antibody provided herein and a pharmaceutically acceptable carrier.

In some embodiments, a method of treating cancer in a mammal is provided, comprising administering an effective amount of an anti-PD-1 antibody provided herein, or a pharmaceutical composition comprising an anti-PD-1 antibody provided herein and a pharmaceutically acceptable carrier. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

In some embodiments, a method of enhancing an immune response in a mammal is provided, comprising administering an effective amount of an anti-PD-1 antibody provided herein, or a pharmaceutical composition comprising an anti-PD-1 antibody provided herein and a pharmaceutically acceptable carrier.

In some embodiments, a method of increasing activation of a T cell in a mammal is provided, comprising administering an effective amount of an anti-PD-1 antibody provided herein, or a pharmaceutical composition comprising an anti-PD-1 antibody provided herein and a pharmaceutically acceptable carrier.

In some embodiments, a method of reducing tumor size in a in a mammal with cancer is provided, comprising administering an effective amount of an anti-PD-1 antibody provided herein, or a pharmaceutical composition comprising an anti-PD-1 antibody provided herein and a pharmaceutically acceptable carrier.

In any of the embodiments provided herein, the mammal may be a human.

In some embodiments, the mammal is administered at least one additional therapeutic agent. In some such embodiments, the additional therapeutic agent is administered concurrently or sequentially with the anti-PD-1 antibody. In some embodiments, the additional therapeutic agent is selected from an anti-ICOS antibody and an anti-CTLA4 antibody. In some embodiments, the additional therapeutic agent is an anti-ICOS antibody. In some embodiments, the additional therapeutic agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from a DNA vaccine, an engineered virus vaccine, an engineered tumor cell vaccine, and a cancer vaccine developed using neoantigens.

In some embodiments, use of an antibody provided herein is provided for the manufacture of a medicament to treat cancer. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC). In some embodiments, the medicament is for administration with at least one additional therapeutic agent. In some embodiments, the additional therapeutic agent is selected from an anti-ICOS antibody and an anti-CTLA4 antibody. In some embodiments, the additional therapeutic agent is an anti-ICOS antibody. In some embodiments, the additional therapeutic agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from a DNA vaccine, an engineered virus vaccine, an engineered tumor cell vaccine, and a cancer vaccine developed using neoantigens.

In some embodiments, the present disclosure provides uses of an antibody provided herein or a pharmaceutical composition comprising an antibody provided herein and a pharmaceutically acceptable carrier for treating cancer. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

In some embodiments, the present disclosure provides uses of an antibody provided herein or a pharmaceutical composition comprising an antibody provided herein and a pharmaceutically acceptable carrier, and at least one additional therapeutic agent, for treating cancer. In some embodiments, the additional therapeutic agent is selected from an anti-ICOS antibody and an anti-CTLA4 antibody. In some embodiments, the additional therapeutic agent is an anti-ICOS antibody. In some embodiments, the additional therapeutic agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from a DNA vaccine, an engineered virus vaccine, an engineered tumor cell vaccine, and a cancer vaccine developed using neoantigens. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

In some embodiments, an antibody provided herein or a pharmaceutical composition comprising an antibody provided herein and a pharmaceutically acceptable carrier for use in treating cancer is provided. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

In some embodiments, a method of increasing the level of at least one cytokine selected from IFNγ and IL-2 in a mammal is provided, comprising administering to the mammal an antibody provided herein. In some embodiments, the level of the cytokine is measured 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 36 hours, or 48 hours after administration of the antibody. In some embodiments, at least one cytokine is IFNγ. In some embodiments, at least one cytokine is IL-2. In some embodiments, the level of the cytokine is measured 24 hours after administration of the antibody. In some embodiments, the mammal is a human. In some embodiments, the human has cancer. In some embodiments, the cancer is selected from melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), gastric cancer, bladder cancer, diffuse large B-cell lymphoma (DLBCL), Hodgkin's lymphoma, ovarian cancer, head & neck squamous cell cancer (HNSCC), mesothelioma, and triple negative breast cancer (TNBC). In some embodiments, the cancer is selected from melanoma, gastric cancer, head & neck squamous cell cancer (HNSCC), non-small cell lung cancer (NSCLC), and triple negative breast cancer (TNBC).

In some embodiments, an immune response is enhanced following administration of the anti-PD-1 antibody. In some embodiments, activation of T cells is increased following administration of the anti-PD-1 antibody. In some embodiments, tumor size is decreased following administration of the anti-PD-1 antibody.

In some embodiments, the mammal is administered at least one additional therapeutic agent. In some embodiments, the additional therapeutic agent is administered concurrently or sequentially with the anti-PD-1 antibody. In some embodiments, the additional therapeutic agent is selected from an anti-ICOS antibody and an anti-CTLA4 antibody. In some embodiments, the additional therapeutic is an anti-ICOS antibody. In some embodiments, the additional therapeutic agent is a cancer vaccine. In some embodiments, the cancer vaccine is selected from a DNA vaccine, an engineered virus vaccine, an engineered tumor cell vaccine, and a cancer vaccine developed using neoantigens.

In some embodiments, a sample of the cancer from the mammal has been determined to express PD-1. In some embodiments, the sample shows 1+, 2+, or 3+ staining of PD-1 by immunohistochemistry (IHC). In some embodiments, the sample has been determined to have an elevated level of PD-L1. In some embodiments, PD-L1 levels are determined using IHC.

