Single domain antibodies to programmed cell death protein 1 (pd-1)

Abstract

The invention relates to PD-1 binding agents that block the interaction of PD-1 with its ligands, and the use of such binding agents in the treatment, prevention and detection of disease.

1 . An isolated V H  single domain antibody that binds to human programmed cell death 1 protein (PD-1) comprising one of the following complementarity determining regions (CDR)1, 2 and 3 combinations: SEQ ID NOs: 1, 2, 3; SEQ ID NOs: 5, 6, 7; SEQ ID NOs: 9, 10, 11; SEQ ID NOs: 13, 14, 15; SEQ ID NOs: 17, 18, 19; SEQ ID NOs: 21, 22, 23; SEQ ID NOs: 25, 26, 27; SEQ ID NOs: 29, 30, 31; SEQ ID NOs: 33, 34, 35; SEQ ID NOs: 37, 38, 39; SEQ ID NOs: 41, 42, 43, SEQ ID NOs: 45, 46, 47; SEQ ID NOs: 49, 50, 51; SEQ ID NOs: 53, 54, 55; SEQ ID NOs: 57, 58, 59; SEQ ID NOs: 61, 62, 63; SEQ ID NOs: 65, 66, 67; SEQ ID NOs: 69, 70, 71; SEQ ID NOs: 73, 74, 75; SEQ ID NOs: 77, 78, 79; SEQ ID NOs: 101, 102, 103; SEQ ID NOs: 105, 106, 107; SEQ ID NOs: 109, 110, 111; SEQ ID NOs: 113, 114, 115; SEQ ID NOs: 117, 118, 119; SEQ ID NOs: 121, 122, 123; SEQ ID NOs: 125, 126, 127; SEQ ID NOs: 129; 130; 131; SEQ ID NOs: 133, 134, 15; SEQ ID NOs: 137, 18, 139; SEQ ID NOs: 141, 142, 143; SEQ ID NOs: 145, 146, 147; SEQ ID NOs: 149, 150, 151; SEQ ID NOs: 153, 154, 155; SEQ ID NOs: 157, 158, 159; SEQ ID NOs: 161, 162, 163; SEQ ID NOs: 165, 166, 167; SEQ ID NOs: 169, 170, 171; SEQ ID NOs: 173, 174, 175; SEQ ID NOs: 177, 178, 179; SEQ ID NOs: 181, 182, 183; SEQ ID NOs: 185, 186, 187; SEQ ID NOs: 189, 190, 191; SEQ ID NOs: 193, 194, 195; SEQ ID NOs: 197, 198, 199; SEQ ID NOs: 201, 202, 203; NOs: 205, 206, 207; SEQ ID NOs: 209, 210, 211; SEQ ID NOs: 213, 214, 215; SEQ ID NOs: 217, 218, 219, SEQ ID NOs: 221, 222, 223, SEQ ID NOs: 225, 226, 227; SEQ ID NOs: 229, 230, 231; SEQ ID NOs: 233, 234, 235; SEQ ID NOs: 237, 238, 239; SEQ ID Nos. 241, 242, 243; SEQ ID NOs: 245, 246, 247; SEQ ID NOs: 249, 250, 251; SEQ ID NOs: 253, 254, 255; SEQ ID NOs: 257, 258, 259; SEQ ID NOs: 261, 262, 263; SEQ ID NOs: 265, 266, 267; SEQ ID NOs: 269, 270, 271; SEQ ID NOs: 273, 274, 275; SEQ ID NOs: 277, 278, 279; SEQ ID NOs: 281, 282, 283; SEQ ID NOs: 285, 286, 287; SEQ ID NOs: 289, 290, 291; SEQ ID NOs: 366, 367, 368; SEQ ID NOs: 370, 371, 372; SEQ ID NOs: 374, 375, 376; SEQ ID NOs: 378, 379, 380; SEQ ID NOs: 382, 383, 384; SEQ ID NOs: 386, 387, 388; SEQ ID NOs: 390, 391, 392; SEQ ID NOs: 394, 395, 396; SEQ ID NOs: 398, 399, 400; SEQ ID NOs: 403, 403, 404; SEQ ID NOs: 406, 407, 408; SEQ ID NOs: 410, 411, 412; SEQ ID NOs: 414, 415, 416; SEQ ID NOs: 418, 419, 420; SEQ ID NOs: 422, 423, 425; SEQ ID NOs: 426, 427, 428; SEQ ID NOs: 430, 431, 432, SEQ ID NOs: 434, 436, 436; SEQ ID NOs: 438, 439, 440; SEQ ID NOs: 442, 443, 444; SEQ ID NOs: 446, 447, 448; SEQ ID NOs: 450, 451, 452; SEQ ID NOs: 454, 455, 456; SEQ ID NOs: 458, 459, 460; SEQ ID NOs: 462, 463, 464; SEQ ID NOs: 466, 467, 468; SEQ ID NOs: 470, 471, 472, SEQ ID NOs: 474, 475, 476; SEQ ID NOs: 478, 479, 480; SEQ ID NOs: 482, 483, 484; SEQ ID NOs: 486, 487, 488; SEQ ID NOs: 490, 491, 492; SEQ ID NOs: 494, 495, 496; SEQ ID NOs: 498, 499, 500; SEQ ID NOs: 502, 503, 504; SEQ ID NOs: 506, 507, 508; SEQ ID NOs: 510, 511, 512; SEQ ID NOs: 514, 515, 516; SEQ ID NOs: 518, 519, 520; SEQ ID NOs: 522, 523, 524; SEQ ID NOs: 526, 527, 528, or SEQ ID NOs: 530, 531, 532. 
     
     
         2 . The isolated V H  single domain antibody of claim comprising a sequence selected from SEQ ID NOs: 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92, 96, 100, 104, 108, 112, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, 156, 160, 164, 168, 172, 176, 180, 184, 188, 192, 196, 200, 204, 208, 212, 216, 220, 224, 228, 232, 236, 240, 244, 248, 252, 256, 260, 264, 268, 272, 276, 280, 284, 288, 292, 369, 373, 377, 381, 385, 389, 393, 397, 401, 405, 409, 413, 417 421, 425, 429, 433, 437, 441, 445, 449, 453, 457, 461, 465, 469, 473, 477, 481, 485, 489, 493, 497, 501, 505, 509, 513, 517, 521, 525, 529 or 533, or a sequence with at least 60%, 70%, 80%, 90% or 95% homology thereto. 
     
     
         3 . The isolated V H  single domain antibody of  claim 1 , wherein said single domain antibody is conjugated to a toxin, enzyme, radioisotope, half-life extending moiety, label, therapeutic molecule or other chemical moiety, optionally wherein said half-life extending moiety is selected from the group consisting of an albumin binding moiety, a transferrin binding moiety, a polyethylene glycol molecule, a recombinant polyethylene glycol molecule, human serum albumin, a fragment of human serum albumin, and an albumin binding peptide or single domain antibody that binds to human serum albumin. 
     
     
         4 . (canceled) 
     
     
         5 . The isolated V H  single domain antibody of  claim 1  obtained or obtainable from a transgenic rodent that expresses a transgene comprising human V, D and J regions, optionally wherein said rodent does not produce any functional endogenous light and heavy chains. 
     
     
         6 . (canceled) 
     
     
         7 . An isolated binding agent that binds to essentially the same epitope as the single domain antibody of  claim 1 . 
     
     
         8 . An isolated binding agent that competes for binding to human PD-1 with the single domain antibody of  claim 1 . 
     
     
         9 . The binding agent of  claim 8 , wherein said binding agent is an antibody or fragment thereof. 
     
     
         10 . The isolated binding agent of  claim 9 , wherein said fragment is a Fab, Fab′, F(ab′)2, dAb, Fd, Fv, a single chain Fv fragment, a human heavy chain variable domain (VH), or an isolated CDR. 
     
     
         11 . An isolated binding agent comprising the single domain antibody of  claim 1 . 
     
     
         12 . The isolated binding agent of  claim 7  wherein said single domain antibody is linked to a second binding molecule that does not bind to PD-1, optionally wherein said second single domain antibody binds to an immuno-oncology target, or wherein said single domain antibody is linked to a second binding molecule that binds to PD-1 at the same, overlapping or different epitope. 
     
     
         13 - 14 . (canceled) 
     
     
         15 . The isolated binding agent of  claim 7 , wherein said binding agent is an antibody or a fragment thereof, optionally wherein said binding molecule is a human heavy chain variable domain (V H ). 
     
     
         16 - 19 . (canceled) 
     
     
         20 . An immunoconjugate comprising the single domain antibody of  claim 1  linked to a therapeutic agent, optionally wherein said therapeutic agent is a toxin, enzyme, radioisotope or other chemical moiety. 
     
     
         21 . (canceled) 
     
     
         22 . A pharmaceutical composition comprising the single domain antibody of  claim 1  and a pharmaceutical carrier. 
     
     
         23 . A method for treating a cancer, an immune disorder, neurological disease, inflammatory disorder, allergy, transplant rejection, viral infection, immune deficiency or other immune system-related disorder, or a method of modulating an immune response, comprising administering a therapeutically effective amount of the single domain antibody of  claim 1 . 
     
     
         24 - 26 . (canceled) 
     
     
         27 . The method of  claim 23 , wherein said cancer is selected from bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, breast cancer, brain cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, kidney cancer, sarcoma of soft tissue, cancer of the urethra, cancer of the bladder, renal cancer, lung cancer, non-small cell lung cancer, thymoma, urothelial carcinoma leukemia, prostate cancer, mesothelioma, adrenocortical carcinoma, lymphomas, such as such as Hodgkin's disease, non-Hodgkin's, gastric cancer, and multiple myelomas. 
     
     
         28 . (canceled) 
     
     
         29 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding the single domain antibody of  claim 1 , optionally wherein the isolated nucleic acid molecule is selected from SEQ ID NOs: 293 to 365 or 534 to 575. 
     
     
         30 . (canceled) 
     
     
         31 . A vector comprising the nucleic acid of  claim 29 . 
     
     
         32 . A host cell comprising the vector of  claim 31 , optionally wherein said host cell is a bacterial, yeast, insect, plant, viral or mammalian cell. 
     
     
         33 . (canceled) 
     
     
         34 . A method for producing said single domain antibody of  claim 1 , comprising expressing a nucleic acid encoding said single domain antibody in a host cell and isolating the single domain antibody from the host cell. 
     
     
         35 . A kit comprising the single domain antibody of  claim 1 . 
     
     
         36 . A method for detecting the presence of human PD-1 in a test sample comprising contacting said sample with the single domain antibody of  claim 1  and at least one detectable label and detecting binding of said single domain antibody to human PD-1. 
     
     
         37 . A method for producing a single V H  domain antibody that binds to human PD-1 comprising
 a) immunising a transgenic animal that expresses a nucleic acid construct comprising human heavy chain V genes and that is not capable of making functional endogenous light or heavy chains with an PD-1 antigen,   b) generating a library from said animal   c) isolating single V H  domain antibodies from said libraries and   d) identifying a single V H  domain antibody that binds to human PD-1 and   e) isolating said antibody.   
     
     
         38 - 39 . (canceled) 
     
     
         40 . A transgenic rodent that produces a heavy chain only antibody that binds to human PD-1, wherein the transgenic rodent expresses a nucleic acid construct comprising human heavy chain V genes, is not capable of making functional endogenous light or heavy chains, and has been immunized with an PD-1 antigen. 
     
     
         41 . (canceled) 
     
     
         42 . A bispecific molecule comprising the single domain antibody of  claim 1  linked to a second functional moiety having a different binding specificity than said single domain antibody, optionally wherein said second moiety is an antibody, an antibody fragment or antibody mimetic, optionally wherein said second moiety binds to an immunooncology target. 
     
     
         43 - 44 . (canceled)