Novel Anti-MYCT1 human antibody and use thereof

Abstract

The present invention relates to an anti-MYCT1 human antibody and a use thereof.

Description

Novel anti-MYCT1 human antibodies and their uses

[0001] The present invention relates to anti-MYCT1 human antibodies and uses thereof.

[0002] The present invention claims priority from Korean Patent Application No. 2023-0173228 filed in the Republic of Korea on December 4, 2023 and Korean Patent Application No. 2024-0177322 filed in the Republic of Korea on December 3, 2024, the contents of which are incorporated herein by reference.

[0003] Angiogenesis is a physiological process in which new blood vessels develop from pre-existing ones. Angiogenesis is known to play a role in both normal and pathological processes. Regulation of angiogenesis is highly regulated by pro- and anti-angiogenic factors, a process that is disrupted and dysregulated in cancer. Tumor-induced hypoxia increases the expression of angiogenic factors, which induce the formation of new blood vessels essential for tumor survival and proliferation. The VEGF family, consisting of six growth factors (VEGFA-F), plays a key role in angiogenesis by binding to the receptors VEGFR1-3 and neuropilin. Angiogenesis can also be mediated independently of the VEGF pathway by the angiopoietin (Ang1-2) / Tie-2 pathway. Accordingly, over the past decade, drug development has focused on anti-angiogenesis as a strategy to deprive tumors of nutrients and inhibit tumor growth. However, despite the weak activity of these agents as single agents or in combination with chemotherapy, tumors still have the problem of overcoming their effects and developing resistance.

[0004] Cancer immunotherapy emerged as an effective treatment for various cancers following the discovery of immune checkpoints. Immune checkpoint inhibitors (ICIs) have demonstrated long-lasting clinical activity against malignant tumors. ICIs activate tumor-fighting effector immune cells by blocking another mechanism hijacked by tumor "immune exhaustion." Primary resistance to ICIs occurs in tumors lacking tumor-infiltrating lymphocytes. Furthermore, tumors that initially respond to ICIs can develop secondary resistance due to defects in antigen presentation mechanisms and overexpression of co-inhibitory molecules.

[0005] Therefore, it is necessary to overcome the problems of VEGF inhibitors and reprogram the tumor microenvironment (TME) to normalize tumor blood vessels and enable immune cells to move well into the TME.

[0006] To achieve this, the development of drugs targeting MYCT1 (MYC Target 1) is necessary. To date, no anti-MYCT1 antibody has been approved as a treatment for human diseases. Therefore, the development of anti-MYCT1 antibodies with strong binding affinity for MYCT1, excellent specificity, potent ADCC and CDC activities, and superior anti-tumor and anti-angiogenic activity through TME reprogramming is required.

[0007] [Prior Art Literature]

[0008] [Non-patent literature]

[0009] J. Cell. Mol. Med., 2016 Vol 20, No 3, pp. 471-481

[0010] An object of the present invention is to provide a human antibody or antigen-binding fragment thereof that binds to MYCT1.

[0011] Another object of the present invention is to provide a nucleic acid molecule encoding the antibody or antigen-binding fragment, an expression vector comprising the same, and a host cell transfected with the vector.

[0012] Another object of the present invention is to provide an immunoconjugate comprising the antibody or antigen-binding fragment and a cytotoxic agent.

[0013] Another object of the present invention is to provide the antibody or antigen-binding fragment for use in the diagnosis or treatment of a MYCT1-related disease.

[0014] Another object of the present invention is to provide a use of the antibody or antigen-binding fragment for diagnosing sensitivity to an immunotherapy agent.

[0015] Another object of the present invention is to provide a chimeric antigen receptor (CAR) comprising the antigen-binding fragment and an immune effector cell expressing the same.

[0016]

[0017] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0018] In order to solve the above problem, one aspect of the present invention is

[0019] A heavy chain CDR1 (HCDR-1) comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 43;

[0020] A heavy chain CDR2 (HCDR-2) comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 44 to 85;

[0021] A heavy chain CDR3 (HCDR-3) comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 187;

[0022] A light chain CDR1 (LCDR-1) comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 188 to 264;

[0023] A light chain CDR2 (LCDR-2) comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 265 to 320; and

[0024] Provided is an anti-MYCT1 human antibody or antigen-binding fragment thereof specific for human MYCT1, comprising a light chain CDR3 (LCDR-3) comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOs: 321 to 411.

[0025] In one specific embodiment, the HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2 and LCDR-3 may each, in order, comprise a CDR combination of the amino acid sequences set forth below:

[0026] SEQ ID NOs: 13, 76, 105, 222, 301, and 346; SEQ ID NOs: 40, 53, 160, 262, 278, and 404;

[0027] SEQ ID NOs: 42, 56, 113, 262, 283, and 407; SEQ ID NOs: 26, 44, 147, 201, 266, and 358;

[0028] SEQ ID NOs: 25, 77, 110, 251, 293, and 388; SEQ ID NOs: 6, 68, 143, 230, 304, and 325;

[0029] SEQ ID NOs: 25, 77, 109, 252, 293, and 382; SEQ ID NOs: 42, 56, 112, 262, 283, and 409;

[0030] SEQ ID NOs: 40, 52, 160, 245, 279, and 401; SEQ ID NOs: 42, 56, 98, 233, 272, and 338;

[0031] SEQ ID NOs: 42, 56, 95, 237, 310, and 324; SEQ ID NOs: 25, 77, 114, 246, 275, and 403;

[0032] SEQ ID NOs: 25, 77, 126, 251, 293, and 386; SEQ ID NOs: 30, 48, 130, 194, 320, and 378;

[0033] SEQ ID NOs: 25, 77, 128, 249, 307, and 328; SEQ ID NOs: 33, 72, 91, 205, 296, and 368;

[0034] SEQ ID NOs: 42, 56, 156, 222, 301, and 346; SEQ ID NOs: 16, 63, 131, 262, 282, and 397;

[0035] SEQ ID NOs: 25, 77, 125, 251, 293, and 384; SEQ ID NOs: 42, 56, 118, 264, 277, and 392;

[0036] SEQ ID NOs: 25, 77, 117, 251, 293, and 387; SEQ ID NOs: 42, 56, 95, 239, 310, and 324;

[0037] SEQ ID NOs: 20, 74, 138, 200, 297, and 354; SEQ ID NOs: 6, 65, 123, 193, 319, and 373;

[0038] SEQ ID NOs: 42, 56, 95, 239, 310, and 322; SEQ ID NOs: 42, 59, 90, 254, 279, and 395;

[0039] SEQ ID NOs: 42, 56, 94, 239, 310, and 325; SEQ ID NOs: 42, 56, 88, 261, 276, and 393;

[0040] SEQ ID NOs: 8, 66, 116, 189, 315, and 340; SEQ ID NOs: 42, 56, 133, 260, 295, and 394;

[0041] SEQ ID NOs: 42, 56, 87, 241, 290, and 334; SEQ ID NOs: 24, 79, 137, 262, 283, and 405;

[0042] SEQ ID NOs: 7, 64, 186, 227, 273, and 342; SEQ ID NOs: 30, 48, 129, 197, 319, and 378;

[0043] SEQ ID NOs: 25, 77, 99, 251, 293, and 387; SEQ ID NOs: 25, 77, 164, 203, 296, and 369;

[0044] SEQ ID NOs: 42, 56, 163, 250, 294, and 383; SEQ ID NOs: 34, 54, 146, 229, 303, and 333;

[0045] SEQ ID NOs: 39, 81, 127, 244, 312, and 335; SEQ ID NOs: 5, 67, 144, 228, 310, and 321;

[0046] SEQ ID NOs: 26, 44, 150, 235, 309, and 330; SEQ ID NOs: 42, 56, 104, 247, 293, and 385;

[0047] SEQ ID NOs: 27, 77, 172, 262, 278, and 408; SEQ ID NOs: 42, 56, 101, 195, 319, and 379;

[0048] SEQ ID NOs: 26, 44, 161, 239, 310, and 324; SEQ ID NOs: 42, 56, 97, 197, 319, and 375;

[0049] SEQ ID NOs: 43, 80, 149, 243, 317, and 323; SEQ ID NOs: 27, 51, 152, 210, 268, and 357;

[0050] SEQ ID NOs: 25, 78, 119, 231, 293, and 387; SEQ ID NOs: 15, 47, 107, 215, 301, and 341;

[0051] SEQ ID NOs: 42, 56, 102, 258, 274, and 402; SEQ ID NOs: 35, 61, 151, 214, 301, and 347;

[0052] SEQ ID NOs: 42, 56, 95, 239, 310, and 325; SEQ ID NOs: 8, 66, 116, 188, 314, and 340;

[0053] SEQ ID NOs: 14, 85, 153, 220, 299, and 349; SEQ ID NOs: 39, 81, 127, 244, 313, and 335;

[0054] SEQ ID NOs: 42, 57, 132, 191, 305, and 389; SEQ ID NOs: 36, 75, 121, 236, 280, and 337;

[0055] SEQ ID NOs: 25, 77, 168, 221, 301, and 345; SEQ ID NOs: 31, 56, 148, 197, 319, and 378;

[0056] SEQ ID NOs: 42, 56, 106, 256, 285, and 396; SEQ ID NOs: 26, 44, 167, 255, 284, and 398;

[0057] SEQ ID NOs: 42, 56, 95, 190, 310, and 324; SEQ ID NOs: 6, 65, 124, 197, 319, and 373;

[0058] SEQ ID NOs: 17, 62, 120, 196, 319, and 380; SEQ ID NOs: 4, 65, 166, 226, 318, and 336;

[0059] SEQ ID NOs: 42, 57, 96, 239, 310, and 325; SEQ ID NOs: 25, 77, 154, 217, 302, and 343;

[0060] SEQ ID NOs: 42, 56, 115, 263, 281, and 411; SEQ ID NOs: 32, 49, 108, 225, 307, and 410;

[0061] SEQ ID NOs: 42, 56, 95, 242, 308, and 331; SEQ ID NOs: 26, 44, 150, 234, 316, and 326;

[0062] SEQ ID NOs: 42, 56, 95, 192, 311, and 322; SEQ ID NOs: 26, 44, 155, 235, 309, and 330;

[0063] SEQ ID NOs: 25, 77, 168, 213, 301, and 345; SEQ ID NOs: 24, 79, 137, 262, 283, and 406;

[0064] SEQ ID NOs: 42, 56, 100, 248, 307, and 391; SEQ ID NOs: 26, 44, 183, 197, 319, and 378;

[0065] SEQ ID NOs: 25, 77, 93, 251, 293, and 387; SEQ ID NOs: 42, 55, 103, 259, 283, and 400;

[0066] SEQ ID NOs: 42, 56, 92, 262, 278, and 399; SEQ ID NOs: 26, 44, 170, 198, 288, and 352;

[0067] SEQ ID NOs: 25, 77, 165, 251, 291, and 387; SEQ ID NOs: 42, 56, 102, 257, 274, and 402;

[0068] SEQ ID NOs: 25, 77, 111, 253, 292, and 381; SEQ ID NOs: 38, 58, 86, 219, 301, and 348;

[0069] SEQ ID NOs: 42, 56, 122, 262, 283, and 405; SEQ ID NOs: 42, 56, 134, 209, 289, and 364;

[0070] SEQ ID NOs: 32, 50, 162, 216, 300, and 344; SEQ ID NOs: 20, 69, 141, 211, 269, and 356;

[0071] SEQ ID NOs: 12, 73, 140, 199, 297, and 353; SEQ ID NOs: 28, 70, 145, 218, 300, and 390;

[0072] SEQ ID NOs: 21, 74, 135, 210, 267, and 370; SEQ ID NOs: 20, 45, 177, 210, 289, and 363;

[0073] SEQ ID NOs: 28, 46, 184, 204, 287, and 366; SEQ ID NOs: 20, 45, 175, 209, 265, and 350;

[0074] SEQ ID NOs: 26, 74, 178, 209, 286, and 377; SEQ ID NOs: 21, 74, 187, 209, 268, and 374;

[0075] SEQ ID NOs: 19, 44, 142, 200, 297, and 355; SEQ ID NOs: 23, 74, 174, 211, 268, and 351;

[0076] SEQ ID NOs: 10, 74, 89, 209, 289, and 372; SEQ ID NOs: 37, 60, 139, 206, 286, and 339;

[0077] SEQ ID NOs: 41, 83, 157, 232, 306, and 329; SEQ ID NOs: 18, 74, 178, 209, 268, and 372;

[0078] SEQ ID NOs: 32, 49, 179, 224, 300, and 344; SEQ ID NOs: 20, 74, 176, 210, 287, and 359;

[0079] SEQ ID NOs: 26, 74, 178, 209, 287, and 361; SEQ ID NOs: 22, 71, 171, 210, 286, and 360;

[0080] SEQ ID NOs: 11, 44, 159, 208, 286, and 371; SEQ ID NOs: 20, 44, 185, 212, 287, and 365;

[0081] SEQ ID NOs: 41, 84, 136, 238, 271, and 332; SEQ ID NOs: 20, 44, 173, 202, 298, and 367;

[0082] SEQ ID NOs: 20, 74, 182, 209, 286, and 376; SEQ ID NOs: 9, 74, 180, 207, 286, and 362;

[0083] SEQ ID NOs: 41, 82, 158, 240, 270, and 327; or SEQ ID NOs: 29, 44, 181, 209, 286, and 376.

[0084] In one embodiment, the antibody or antigen-binding fragment thereof may comprise at least one variable heavy chain region (VH) selected from the group consisting of SEQ ID NOs: 412 to 527.

[0085] In another embodiment, the antibody or antigen-binding fragment thereof may comprise at least one variable light chain region (VL) selected from the group consisting of SEQ ID NOs: 528 to 643.

[0086] In another specific embodiment, the antibody or antigen-binding fragment thereof,

[0087] Containing the VH sequence of SEQ ID NO: 412 and the VL sequence of SEQ ID NO: 528; or

[0088] Containing the VH sequence of SEQ ID NO: 413 and the VL sequence of SEQ ID NO: 529; or

[0089] Containing the VH sequence of SEQ ID NO: 414 and the VL sequence of SEQ ID NO: 530;

[0090] Containing the VH sequence of SEQ ID NO: 415 and the VL sequence of SEQ ID NO: 531;

[0091] Containing the VH sequence of SEQ ID NO: 416 and the VL sequence of SEQ ID NO: 532;

[0092] Containing the VH sequence of SEQ ID NO: 417 and the VL sequence of SEQ ID NO: 533;

[0093] Containing the VH sequence of SEQ ID NO: 418 and the VL sequence of SEQ ID NO: 534;

[0094] Containing the VH sequence of SEQ ID NO: 419 and the VL sequence of SEQ ID NO: 535;

[0095] Containing the VH sequence of SEQ ID NO: 420 and the VL sequence of SEQ ID NO: 536;

[0096] Containing the VH sequence of SEQ ID NO: 421 and the VL sequence of SEQ ID NO: 537;

[0097] Containing the VH sequence of SEQ ID NO: 422 and the VL sequence of SEQ ID NO: 538;

[0098] Containing the VH sequence of SEQ ID NO: 423 and the VL sequence of SEQ ID NO: 539; or

[0099] Containing the VH sequence of SEQ ID NO: 424 and the VL sequence of SEQ ID NO: 540;

[0100] Containing the VH sequence of SEQ ID NO: 425 and the VL sequence of SEQ ID NO: 541; or

[0101] Containing the VH sequence of SEQ ID NO: 426 and the VL sequence of SEQ ID NO: 542;

[0102] Containing the VH sequence of SEQ ID NO: 427 and the VL sequence of SEQ ID NO: 543;

[0103] Containing the VH sequence of SEQ ID NO: 428 and the VL sequence of SEQ ID NO: 544;

[0104] Containing the VH sequence of SEQ ID NO: 429 and the VL sequence of SEQ ID NO: 545; or

[0105] Containing the VH sequence of SEQ ID NO: 430 and the VL sequence of SEQ ID NO: 546;

[0106] Containing the VH sequence of SEQ ID NO: 431 and the VL sequence of SEQ ID NO: 547; or

[0107] Containing the VH sequence of SEQ ID NO: 432 and the VL sequence of SEQ ID NO: 548; or

[0108] Contains the VH sequence of SEQ ID NO: 433 and the VL sequence of SEQ ID NO: 549; or

[0109] It may include the VH sequence of SEQ ID NO: 434 and the VL sequence of SEQ ID NO: 550.

[0110]

[0111] In another specific embodiment, the antibody or antigen-binding fragment thereof,

[0112] Containing the VH sequence of SEQ ID NO: 435 and the VL sequence of SEQ ID NO: 551;

[0113] Containing the VH sequence of SEQ ID NO: 436 and the VL sequence of SEQ ID NO: 552;

[0114] Containing the VH sequence of SEQ ID NO: 437 and the VL sequence of SEQ ID NO: 553;

[0115] Containing the VH sequence of SEQ ID NO: 438 and the VL sequence of SEQ ID NO: 554;

[0116] Containing the VH sequence of SEQ ID NO: 439 and the VL sequence of SEQ ID NO: 555;

[0117] Containing the VH sequence of SEQ ID NO: 440 and the VL sequence of SEQ ID NO: 556;

[0118] Containing the VH sequence of SEQ ID NO: 441 and the VL sequence of SEQ ID NO: 557;

[0119] Containing the VH sequence of SEQ ID NO: 442 and the VL sequence of SEQ ID NO: 558; or

[0120] Containing the VH sequence of SEQ ID NO: 443 and the VL sequence of SEQ ID NO: 559;

[0121] Containing the VH sequence of SEQ ID NO: 444 and the VL sequence of SEQ ID NO: 560;

[0122] Containing the VH sequence of SEQ ID NO: 445 and the VL sequence of SEQ ID NO: 561;

[0123] Containing the VH sequence of SEQ ID NO: 446 and the VL sequence of SEQ ID NO: 562;

[0124] Containing the VH sequence of SEQ ID NO: 447 and the VL sequence of SEQ ID NO: 563;

[0125] Containing the VH sequence of SEQ ID NO: 448 and the VL sequence of SEQ ID NO: 564;

[0126] Containing the VH sequence of SEQ ID NO: 449 and the VL sequence of SEQ ID NO: 565;

[0127] Containing the VH sequence of SEQ ID NO: 450 and the VL sequence of SEQ ID NO: 566;

[0128] Containing the VH sequence of SEQ ID NO: 451 and the VL sequence of SEQ ID NO: 567;

[0129] Containing the VH sequence of SEQ ID NO: 452 and the VL sequence of SEQ ID NO: 568;

[0130] Containing the VH sequence of SEQ ID NO: 453 and the VL sequence of SEQ ID NO: 569; or

[0131] Containing the VH sequence of SEQ ID NO: 454 and the VL sequence of SEQ ID NO: 570;

[0132] Containing the VH sequence of SEQ ID NO: 455 and the VL sequence of SEQ ID NO: 571;

[0133] Containing the VH sequence of SEQ ID NO: 456 and the VL sequence of SEQ ID NO: 572;

[0134] Containing the VH sequence of SEQ ID NO: 457 and the VL sequence of SEQ ID NO: 573;

[0135] Containing the VH sequence of SEQ ID NO: 458 and the VL sequence of SEQ ID NO: 574; or

[0136] Containing the VH sequence of SEQ ID NO: 459 and the VL sequence of SEQ ID NO: 575;

[0137] Containing the VH sequence of SEQ ID NO: 460 and the VL sequence of SEQ ID NO: 576;

[0138] Containing the VH sequence of SEQ ID NO: 461 and the VL sequence of SEQ ID NO: 577;

[0139] Containing the VH sequence of SEQ ID NO: 462 and the VL sequence of SEQ ID NO: 578;

[0140] Containing the VH sequence of SEQ ID NO: 463 and the VL sequence of SEQ ID NO: 579; or

[0141] Containing the VH sequence of SEQ ID NO: 464 and the VL sequence of SEQ ID NO: 580;

[0142] Containing the VH sequence of SEQ ID NO: 465 and the VL sequence of SEQ ID NO: 581;

[0143] Containing the VH sequence of SEQ ID NO: 466 and the VL sequence of SEQ ID NO: 582;

[0144] Containing the VH sequence of SEQ ID NO: 467 and the VL sequence of SEQ ID NO: 583;

[0145] Containing the VH sequence of SEQ ID NO: 468 and the VL sequence of SEQ ID NO: 584;

[0146] Containing the VH sequence of SEQ ID NO: 469 and the VL sequence of SEQ ID NO: 585;

[0147] Containing the VH sequence of SEQ ID NO: 470 and the VL sequence of SEQ ID NO: 586;

[0148] Containing the VH sequence of SEQ ID NO: 471 and the VL sequence of SEQ ID NO: 587;

[0149] Containing the VH sequence of SEQ ID NO: 472 and the VL sequence of SEQ ID NO: 588; or

[0150] Containing the VH sequence of SEQ ID NO: 473 and the VL sequence of SEQ ID NO: 589; or

[0151] Containing the VH sequence of SEQ ID NO: 474 and the VL sequence of SEQ ID NO: 590;

[0152] Containing the VH sequence of SEQ ID NO: 475 and the VL sequence of SEQ ID NO: 591; or

[0153] Containing the VH sequence of SEQ ID NO: 476 and the VL sequence of SEQ ID NO: 592;

[0154] Containing the VH sequence of SEQ ID NO: 477 and the VL sequence of SEQ ID NO: 593;

[0155] Containing the VH sequence of SEQ ID NO: 478 and the VL sequence of SEQ ID NO: 594;

[0156] Containing the VH sequence of SEQ ID NO: 479 and the VL sequence of SEQ ID NO: 595; or

[0157] Containing the VH sequence of SEQ ID NO: 480 and the VL sequence of SEQ ID NO: 596;

[0158] Containing the VH sequence of SEQ ID NO: 481 and the VL sequence of SEQ ID NO: 597; or

[0159] Containing the VH sequence of SEQ ID NO: 482 and the VL sequence of SEQ ID NO: 598; or

[0160] Containing the VH sequence of SEQ ID NO: 483 and the VL sequence of SEQ ID NO: 599;

[0161] Containing the VH sequence of SEQ ID NO: 484 and the VL sequence of SEQ ID NO: 600;

[0162] Containing the VH sequence of SEQ ID NO: 485 and the VL sequence of SEQ ID NO: 601;

[0163] Containing the VH sequence of SEQ ID NO: 486 and the VL sequence of SEQ ID NO: 602;

[0164] Containing the VH sequence of SEQ ID NO: 487 and the VL sequence of SEQ ID NO: 603;

[0165] Containing the VH sequence of SEQ ID NO: 488 and the VL sequence of SEQ ID NO: 604;

[0166] Containing the VH sequence of SEQ ID NO: 489 and the VL sequence of SEQ ID NO: 605;

[0167] Containing the VH sequence of SEQ ID NO: 490 and the VL sequence of SEQ ID NO: 606;

[0168] Containing the VH sequence of SEQ ID NO: 491 and the VL sequence of SEQ ID NO: 607; or

[0169] Containing the VH sequence of SEQ ID NO: 492 and the VL sequence of SEQ ID NO: 608;

[0170] Containing the VH sequence of SEQ ID NO: 493 and the VL sequence of SEQ ID NO: 609;

[0171] Containing the VH sequence of SEQ ID NO: 494 and the VL sequence of SEQ ID NO: 610;

[0172] Containing the VH sequence of SEQ ID NO: 495 and the VL sequence of SEQ ID NO: 611;

[0173] Containing the VH sequence of SEQ ID NO: 496 and the VL sequence of SEQ ID NO: 612;

[0174] Containing the VH sequence of SEQ ID NO: 497 and the VL sequence of SEQ ID NO: 613;

[0175] Containing the VH sequence of SEQ ID NO: 498 and the VL sequence of SEQ ID NO: 614;

[0176] Containing the VH sequence of SEQ ID NO: 499 and the VL sequence of SEQ ID NO: 615;

[0177] Containing the VH sequence of SEQ ID NO: 500 and the VL sequence of SEQ ID NO: 616;

[0178] Containing the VH sequence of SEQ ID NO: 501 and the VL sequence of SEQ ID NO: 617; or

[0179] Containing the VH sequence of SEQ ID NO: 502 and the VL sequence of SEQ ID NO: 618; or

[0180] Containing the VH sequence of SEQ ID NO: 503 and the VL sequence of SEQ ID NO: 619; or

[0181] Containing the VH sequence of SEQ ID NO: 504 and the VL sequence of SEQ ID NO: 620;

[0182] Containing the VH sequence of SEQ ID NO: 505 and the VL sequence of SEQ ID NO: 621; or

[0183] Containing the VH sequence of SEQ ID NO: 506 and the VL sequence of SEQ ID NO: 622;

[0184] Containing the VH sequence of SEQ ID NO: 507 and the VL sequence of SEQ ID NO: 623;

[0185] Containing the VH sequence of SEQ ID NO: 508 and the VL sequence of SEQ ID NO: 624;

[0186] Containing the VH sequence of SEQ ID NO: 509 and the VL sequence of SEQ ID NO: 625;

[0187] Containing the VH sequence of SEQ ID NO: 510 and the VL sequence of SEQ ID NO: 626;

[0188] Containing the VH sequence of SEQ ID NO: 511 and the VL sequence of SEQ ID NO: 627;

[0189] Containing the VH sequence of SEQ ID NO: 512 and the VL sequence of SEQ ID NO: 628;

[0190] Containing the VH sequence of SEQ ID NO: 513 and the VL sequence of SEQ ID NO: 629;

[0191] Containing the VH sequence of SEQ ID NO: 514 and the VL sequence of SEQ ID NO: 630;

[0192] Containing the VH sequence of SEQ ID NO: 515 and the VL sequence of SEQ ID NO: 631;

[0193] Containing the VH sequence of SEQ ID NO: 516 and the VL sequence of SEQ ID NO: 632;

[0194] Containing the VH sequence of SEQ ID NO: 517 and the VL sequence of SEQ ID NO: 633;

[0195] Containing the VH sequence of SEQ ID NO: 518 and the VL sequence of SEQ ID NO: 634;

[0196] Containing the VH sequence of SEQ ID NO: 519 and the VL sequence of SEQ ID NO: 635; or

[0197] Containing the VH sequence of SEQ ID NO: 520 and the VL sequence of SEQ ID NO: 636;

[0198] Containing the VH sequence of SEQ ID NO: 521 and the VL sequence of SEQ ID NO: 637;

[0199] Containing the VH sequence of SEQ ID NO: 522 and the VL sequence of SEQ ID NO: 638;

[0200] Containing the VH sequence of SEQ ID NO: 523 and the VL sequence of SEQ ID NO: 639;

[0201] Containing the VH sequence of SEQ ID NO: 524 and the VL sequence of SEQ ID NO: 640;

[0202] Containing the VH sequence of SEQ ID NO: 525 and the VL sequence of SEQ ID NO: 641;

[0203] Contains the VH sequence of SEQ ID NO: 526 and the VL sequence of SEQ ID NO: 642; or

[0204] It may include the VH sequence of SEQ ID NO: 527 and the VL sequence of SEQ ID NO: 643.

[0205] The above antigen-binding fragments may include, but are not limited to, Fd, Fab, Fab', F(ab')2, dsFv, scFv, and single domain antibodies (sdAb), and any fragment that possesses at least antigen-binding property may be used without limitation.

[0206] In another aspect, the present invention provides a nucleic acid molecule encoding the antibody or antigen-binding fragment thereof.

[0207] In another aspect, the present invention provides a recombinant expression vector comprising the nucleic acid molecule; and a host cell transfected with the expression vector.

[0208] In another aspect, the present invention provides an immunoconjugate comprising the antibody or antigen-binding fragment and a cytotoxic agent.

[0209] In another aspect, the present invention provides a pharmaceutical composition for preventing or treating a MYCT1-related disease, comprising the antibody or antigen-binding fragment as an active ingredient.

[0210] In one specific example, the MYCT1-related disease may be an angiogenesis-related disease or cancer.

[0211] In other embodiments, the pharmaceutical composition may be administered in combination with at least one additional therapeutic agent or therapeutic procedure.

[0212] The at least one additional therapeutic agent or therapeutic procedure may be selected from one or more of chemotherapy, targeted anticancer therapy, oncolytic drug, cytotoxic agent, immune-based therapy, cytokine, surgical procedure, radiation procedure, vaccine, or cell therapy.

[0213]

[0214] In another aspect, the present invention provides a composition for diagnosing a MYCT1-related disease comprising the antibody or an antigen-binding fragment thereof as an active ingredient.

[0215] In another aspect, the present invention provides a pharmaceutical composition for enhancing sensitivity to an immunotherapy agent in cancer, comprising the antibody or an antigen-binding fragment thereof as an active ingredient.

[0216] In another aspect, the present invention

[0217] A chimeric antigen receptor (CAR) comprising an antigen binding domain that specifically binds to MYCT1 is provided. Specifically, the antigen binding domain comprises:

[0218] A heavy chain CDR1 (HCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 43;

[0219] A heavy chain CDR2 (HCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 44 to 85;

[0220] A heavy chain CDR3 (HCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 187;

[0221] A light chain CDR1 (LCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 188 to 264;

[0222] A light chain CDR2 (LCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 265 to 320; and

[0223] A light chain CDR3 (LCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 321 to 411;

[0224] In another aspect, the present invention provides an immune effector cell comprising the chimeric antigen receptor.

[0225] In another aspect, the present invention provides a method for treating a MYCT1-associated disease comprising administering to a subject a composition comprising a therapeutically effective amount of the antibody or antigen-binding fragment.

[0226] In another aspect, the present invention provides a method for enhancing the sensitivity of a subject to an immunotherapy agent, comprising administering to the subject a composition comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof.

[0227] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0228] The human antibody or antigen-binding fragment thereof that specifically binds to MYCT1 provided in the present invention exhibits excellent affinity and binding ability to MYCT1, and thus, an antibody-drug conjugate, a bispecific antibody, or a chimeric antigen receptor (CAR) comprising the antibody or antigen-binding fragment can be developed. In addition, the antibody or antigen-binding fragment according to the present invention can be usefully applied to the treatment or diagnosis of MYCT1-related diseases, such as cancer or angiogenesis-related diseases. Furthermore, the antibody or antigen-binding fragment according to the present invention can be used in combination with an immunotherapy agent and can enhance sensitivity to the immunotherapy agent.

[0229]

[0230] The effects of the composition according to one embodiment of the present invention are not limited to those mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description below.

[0231] Figure 1 compares the MYCT1 sequences of humans and mice.

[0232] Figures 2a, 2b, 2c, 2d and Figures 3a, 3b, 3c, 3d show the results of measuring the viable cell density and viability of ExpiCHO-S cells transformed with the anti-MYCT1 gene.

[0233] Figure 4 shows the results of Protein A HPLC titer measurement of anti-MYCT1 antibody.

[0234] Figures 5a, 5b, 5c, 5d, 5e and 5f are Protein A FPLC chromatograms of anti-MYCT1 antibody.

[0235] Figures 6a, 6b, 6c, 6d, Figures 7a, 7b, 7c, 7d and Figure 8 are the results of confirming the purity (SDS-PAGE) of an anti-MYCT1 antibody according to one embodiment of the present invention.

[0236] Figures 9a, 9b and 9c are the results of antigen binding specificity analysis (ELISA) of an anti-MYCT1 antibody according to one embodiment of the present invention.

[0237] Figures 10a, 10b, 10c, 10d, 10e, 10f and 10g are cell binding specificity analysis (FACS) results of anti-MYCT1 antibodies according to one embodiment of the present invention.

[0238] Figures 11a, 11b and 11c show the results of SPR overall dynamics analysis of an anti-MYCT1 antibody according to one embodiment of the present invention.

[0239] Figures 12a, 12b and 13a, 13b show the results of angiogenesis inhibition efficacy evaluation (Migration Assay) of an anti-MYCT1 antibody according to one embodiment of the present invention.

[0240] The antibody or antigen-binding fragment according to the present invention can overcome the limitations of existing immuno-oncology agents by specifically binding to MYCT1 and ultimately reprogramming the tumor microenvironment (TME) through i) inhibition of angiogenesis, ii) normalization of blood vessels, iii) increase in immune cell activity within the TME, and iv) increase infiltration of immune cells into the TME.

[0241] The above MYCT1 (hMYCT1) sequence is as follows (SEQ ID NO: 1).

[0242] MRTQVYEGLCKNYFSLAVLQRDRIKLLFFDILVFLSVFLLFLLFLLVDIMANNTTSLGSPWPENFWEDLIMSFTVSMAIGLVLGGFIWAVFICLSRRRRASAPISQWSSSRRSRSSYT HGLNRTGFYRHSGCERRSNLSLASLTFQRQASLEQANSFPRKSSFRASTFHPFLQCPPLPVETESQLVTLPSSNISPTISTSHSLSRPDYWSSNSLRVGLSTPPPPAYESIIKAFPDS

[0243] The above sequence number 1 is an example of a MYCT1 sequence, and all known MYCT1 sequences, such as the mouse MYCT1 sequence (SEQ ID NO: 2), etc., can be included in the present invention. The human antibody that specifically binds to MYCT1 provided in the present invention can specifically bind to the amino acid sequence from position 49 to position 67 of the above sequence (MANNTTSLGSPWPENFWED, SEQ ID NO: 3).

[0244]

[0245] antibody or antigen-binding fragment

[0246] As used herein, the term "antibody" refers to a protein molecule that acts as a ligand that specifically recognizes an antigen, including an immunoglobulin molecule that immunologically has reactivity with a specific antigen, and includes polyclonal antibodies, monoclonal antibodies, whole antibodies, and antibody fragments. In addition, the antibody in the present invention includes a chimeric antibody, a bivalent antibody, a bispecific molecule, a minibody, a domain antibody, a bispecific or multispecific antibody, an immune cell engaging bispecific or multispecific antibody, an antibody mimetic, a unibody, a diabody, a triabody, or a tetrabody. The term further includes a single-chain antibody having a binding function to FcRn (neonatal Fc receptor), a scab, a derivative of an antibody constant region, and an artificial antibody based on a protein scaffold. A whole antibody has a structure having two full-length light chains (LC) and two full-length heavy chains (HC), each light chain being linked to a heavy chain by a disulfide bond. The whole antibody includes IgA, IgD, IgE, IgM, and IgG, and IgG is a subtype that includes IgG1, IgG2, IgG3, and IgG4. In one embodiment, the antibody provided by the present invention may be an IgG antibody.

[0247] The term "human antibody" in the present invention refers to a molecule derived from human immunoglobulin, in which the entire amino acid sequence constituting the antibody, including the complementarity determining regions and structural domains, is comprised of the amino acid sequence of human immunoglobulin. Human antibodies are commonly used to treat human diseases, and may have the following advantages. First, they interact better with the human immune system, allowing them to more efficiently destroy target cells, for example, through complement-dependent cytotoxicity (CDC) or antibody-dependent cell-mediated cytotoxicity (ADCC). Second, they have the advantage that the human immune system does not recognize the antibody as foreign. Third, they have the advantage of having a half-life in the human circulation similar to that of naturally occurring antibodies, even when administered in smaller amounts or less frequently.

[0248] In addition, when the antibody of the present invention as described above includes a constant region, it may include a constant region derived from IgG, IgA, IgD, IgE, IgM, or a combination thereof or a hybrid thereof.

[0249] The antibody of the present invention can be easily manufactured using known antibody manufacturing techniques. For example, the method for manufacturing a monoclonal antibody can be performed by manufacturing a hybridoma using B lymphocytes obtained from an immunized animal (Koeher and Milstein, 1976, Nature, 256:495), or by utilizing phage display technology, etc., but is not limited thereto, and can be easily manufactured using other known antibody manufacturing techniques.

[0250] An antibody library using phage display technology is a method of directly obtaining antibody genes from B lymphocytes and expressing antibodies on the surface of phage without producing hybridomas. Phage display technology can overcome many of the difficulties associated with producing monoclonal antibodies through B-cell immortalization. In general, phage display technology includes the steps of: 1) inserting an oligonucleotide with a random sequence into the gene region corresponding to the N-terminus of the phage coat protein pIII (or pIV); 2) expressing a fusion protein of a portion of a native coat protein and a polypeptide encoded by the oligonucleotide with a random sequence; 3) treating a substance capable of binding to the polypeptide encoded by the oligonucleotide; 4) eluting antibody-phage particles bound to the substance using low pH or a binding-competitive molecule; 5) amplifying the phage eluted by panning in a host cell; 6) repeating the above method to obtain a desired amount; and 7) determining the sequence of an active antibody from the DNA sequence of phage clones selected by panning.

[0251] The method for producing a monoclonal antibody of the present invention can be performed using phage display technology. Those skilled in the art can easily perform each step of the production method of the present invention by referring to known phage display technologies, for example, methods known in the papers of Barbas et al. (METHODS: A Companion to Methods in Enzymology 2:119, 1991 and J. Virol. 2001 Jul; 75(14):6692-9) and Winter et al. (Ann. Rev. Immunol. 12:433, 1994). Phages that can be used to construct an antibody library include, but are not limited to, filamentous phages such as fd, M13, f1, If1, Ike, Zj / Z, Ff, Xf, Pf1 or Pf3 phages. In addition, vectors that can be used for expression of heterologous genes on the surface of the filamentous phage include, but are not limited to, phage vectors such as fUSE5, fAFF1, fd-CAT1, or fdtetDOG, or phagemid vectors such as pHEN1, pComb3, pComb8, or pSEX. In addition, helper phage that can be used to provide the wild-type coat protein required for successful reinfection of the recombinant phage for amplification include, but are not limited to, M13K07 or VSCM13.

[0252] The polynucleotide encoding the monoclonal antibody or phage display clone of the present invention can be readily isolated and sequenced using conventional procedures. For example, oligonucleotide primers designed to specifically amplify the heavy and light chain coding regions from phage template DNA can be used. Once the polynucleotide is isolated, it can be placed into an expression vector, and the expression vector can then be introduced into a suitable host cell, thereby producing the desired monoclonal antibody from the transformed host cell (i.e., a transformant). Accordingly, the method for producing the human monoclonal antibody of the present invention may be, but is not limited to, a method for producing a human monoclonal antibody comprising a step of amplifying a polynucleotide encoding the human monoclonal antibody from an expression vector containing the polynucleotide encoding the human monoclonal antibody.

[0253] In the present invention, "humanized antibody" means an antibody that exhibits reduced immunogenicity or is non-immunogenic in humans. The humanized antibody may be produced by, for example, combining a CDR derived from a non-human organism (non-human species) with a constant region derived from a human antibody or a framework region (FR) among the variable regions derived from a human antibody. The humanized antibody may be produced by grafting a CDR of a non-human species antibody between the FR sequences of a human antibody through a CDR-grafting method.

[0254] As used herein, the term "monoclonal antibody" refers to an antibody molecule of a single molecular composition obtained from a substantially identical antibody population, and such monoclonal antibody exhibits a single binding specificity and affinity for a specific epitope. Typically, immunoglobulins have heavy and light chains, and each heavy and light chain comprises a constant region and a variable region (also known as a domain). The variable regions of the light and heavy chains comprise three variable regions and four structural regions, designated as complementarity-determining regions (CDRs).

[0255] The term "complementarity determining region" or "CDR region" or "CDR" as used herein refers to a highly variable region of sequence within an antibody variable region, which forms structurally defined loops ("hypervariable loops") and / or comprises antigen contact residues ("antigen contact sites"). CDRs are primarily responsible for binding to antigenic epitopes. The complementarity determining regions are sandwiched between relatively conserved regions called constant regions (FRs). Each heavy chain variable region (VH) and light chain variable region (VL) consists of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The CDRs of the heavy chain variable region may be referred to as HCDR-1, HCDR-2, HCDR-3, the CDRs of the light chain variable region may be referred to as LCDR-1, LCDR-2, LCDR-3, the FRs of the heavy chain variable region may be referred to as HFR1, HFR2, HFR3, HFR4, and the FRs of the light chain variable region may be referred to as LFR1, LFR2, LFR3, LFR4. The numbers are sequentially numbered from the N-terminus. The boundaries of the exact amino acid sequence of each CDR in a given light chain variable region and heavy chain variable region amino acid sequence can be determined, for example, by Chothia (Chothia et al. (1989) Nature 342: 877-883; Al-Lazikani et al., "Standard conformations for the canonical structures of immunoglobulins", Journal of Molecular Biology, 273, 927-948 (1997)), Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, 4th ed., US Department of Health and Human Services, National Institutes of Health (1987)) based on antibody sequence variability, AbM (University of Bath), Contact (University College London), International ImMunoGeneTics database (IMGT) (imgt.cines.fr / on the World Wide Web), and North CDR definition based on affinity propagation clustering using multiple crystal structures, or a combination thereof.

[0256] The antibody of the present invention may comprise a constant region, wherein the constant region may be, but is not limited to, a constant region derived from IgG, IgA, IgD, IgE, IgM, or a combination thereof or a hybrid thereof. Here, the term "combination" means that a polypeptide encoding a single-chain immunoglobulin constant region of the same origin forms a bond with a single-chain polypeptide of a different origin when forming a dimer or multimer. For example, a dimer or multimer may be formed from two or more constant regions selected from the group consisting of constant regions of IgG, IgA, IgD, IgE, and IgM. Additionally, the term “hybrid” or “hybrid” means that within a single-chain immunoglobulin heavy chain constant region, there are sequences corresponding to immunoglobulin heavy chain constant regions of two or more different origins, for example, a hybrid of domains consisting of one to four domains selected from the group consisting of CH1, CH2, CH3 and CH4 of IgG, IgA, IgD, IgE and IgM is possible.

[0257] The term "full-length antibody" as used herein refers to a structure having two full-length light chains and two full-length heavy chains, each light chain being linked to a heavy chain by a disulfide bond, and includes IgA, IgD, IgE, IgM, and IgG. The IgG subtypes include IgG1, IgG2, IgG3, and IgG4.

[0258] As used herein, the terms "fragment," "binding fragment of polypeptide," and "antibody fragment" are used interchangeably to refer to any fragment of an antibody of the present invention that retains the antigen-binding activity of the antibody. Exemplary antibody fragments include, but are not limited to, Fd, Fab, Fab', F(ab')2, dsFv, scFv, or single domain antibodies (sdAb). The Fd refers to the heavy chain portion included in the Fab fragment. As used herein, the term "Fab" typically refers to a fragment comprising a heavy chain variable domain and a light chain variable domain, and further comprising a light chain constant domain and a heavy chain first constant domain (CH1). The term "Fab" typically refers to a fragment that differs from a Fab by the addition of a few residues (including one or more cysteines from the antibody hinge region) to the carboxy terminus of the heavy chain CH1 domain. The term "F(ab')2" refers to a dimeric antibody fragment, typically a Fab', comprising two Fab fragments linked by a disulfide bridge at the hinge region. The term "Fv" typically refers to the smallest antibody fragment containing a complete antigen recognition and binding site. In certain cases, the fragment may be configured as a dimer in which the heavy chain variable region and the light chain variable region are closely non-covalently associated. The term "dsFv" typically refers to an Fv fragment in which the disulfide bond is stabilized, wherein the linkage between a single light chain variable region and a single heavy chain variable region is a disulfide bond. The term "dAb fragment" typically refers to an antibody fragment consisting of a VH domain. In the present invention, the term "scFv" typically refers to a monovalent molecule formed by covalent pairing of the heavy chain variable domain and the light chain variable domain of an antibody via a flexible peptide linker. These scFv molecules can have the general structure NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH.The term "single domain antibody" refers to antibody fragments known as nanobodies (e.g., sdAb, sdFv, nanobody) and composed of a single monomeric variable antibody domain.

[0259] The terms "human anti-MYCT1 antibody", "anti-MYCT1 human antibody", "anti-MYCT1", "MYCT1 antibody" or "antibody that binds to MYCT1" as used herein refer to a human antibody capable of binding to MYCT1 with sufficient affinity such that the antibody can be used as a therapeutic agent targeting MYCT1. In one embodiment of the present invention, the anti-MYCT1 human antibody binds to MYCT1 with high affinity in vitro or in vivo. Here, the binding can be determined by, for example, but not limited to, radioimmunoassay (RIA), biolayer interferometry (BLI), MSD analysis, surface plasmon resonance (SPR) or flow cytometry.

[0260]

[0261] The anti-MYCT1 human antibody or antigen-binding fragment thereof of the present invention

[0262] A heavy chain CDR1 (HCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 43;

[0263] A heavy chain CDR2 (HCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 44 to 85;

[0264] A heavy chain CDR3 (HCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 187;

[0265] A light chain CDR1 (LCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 188 to 264;

[0266] A light chain CDR2 (LCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 265 to 320; and

[0267] It may include one amino acid sequence selected from the group consisting of sequence numbers 321 to 411.

[0268]

[0269] In another specific embodiment, the HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2 and LCDR-3 may each, in order, comprise a CDR combination of the amino acid sequences set forth below:

[0270] SEQ ID NOs: 13, 76, 105, 222, 301, and 346; SEQ ID NOs: 40, 53, 160, 262, 278, and 404;

[0271] SEQ ID NOs: 42, 56, 113, 262, 283, and 407; SEQ ID NOs: 26, 44, 147, 201, 266, and 358;

[0272] SEQ ID NOs: 25, 77, 110, 251, 293, and 388; SEQ ID NOs: 6, 68, 143, 230, 304, and 325;

[0273] SEQ ID NOs: 25, 77, 109, 252, 293, and 382; SEQ ID NOs: 42, 56, 112, 262, 283, and 409;

[0274] SEQ ID NOs: 40, 52, 160, 245, 279, and 401; SEQ ID NOs: 42, 56, 98, 233, 272, and 338;

[0275] SEQ ID NOs: 42, 56, 95, 237, 310, and 324; SEQ ID NOs: 25, 77, 114, 246, 275, and 403;

[0276] SEQ ID NOs: 25, 77, 126, 251, 293, and 386; SEQ ID NOs: 30, 48, 130, 194, 320, and 378;

[0277] SEQ ID NOs: 25, 77, 128, 249, 307, and 328; SEQ ID NOs: 33, 72, 91, 205, 296, and 368;

[0278] SEQ ID NOs: 42, 56, 156, 222, 301, and 346; SEQ ID NOs: 16, 63, 131, 262, 282, and 397;

[0279] SEQ ID NOs: 25, 77, 125, 251, 293, and 384; SEQ ID NOs: 42, 56, 118, 264, 277, and 392;

[0280] SEQ ID NOs: 25, 77, 117, 251, 293, and 387; SEQ ID NOs: 42, 56, 95, 239, 310, and 324;

[0281] SEQ ID NOs: 20, 74, 138, 200, 297, and 354; SEQ ID NOs: 6, 65, 123, 193, 319, and 373;

[0282] SEQ ID NOs: 42, 56, 95, 239, 310, and 322; SEQ ID NOs: 42, 59, 90, 254, 279, and 395;

[0283] SEQ ID NOs: 42, 56, 94, 239, 310, and 325; SEQ ID NOs: 42, 56, 88, 261, 276, and 393;

[0284] SEQ ID NOs: 8, 66, 116, 189, 315, and 340; SEQ ID NOs: 42, 56, 133, 260, 295, and 394;

[0285] SEQ ID NOs: 42, 56, 87, 241, 290, and 334; SEQ ID NOs: 24, 79, 137, 262, 283, and 405;

[0286] SEQ ID NOs: 7, 64, 186, 227, 273, and 342; SEQ ID NOs: 30, 48, 129, 197, 319, and 378;

[0287] SEQ ID NOs: 25, 77, 99, 251, 293, and 387; SEQ ID NOs: 25, 77, 164, 203, 296, and 369;

[0288] SEQ ID NOs: 42, 56, 163, 250, 294, and 383; SEQ ID NOs: 34, 54, 146, 229, 303, and 333;

[0289] SEQ ID NOs: 39, 81, 127, 244, 312, and 335; SEQ ID NOs: 5, 67, 144, 228, 310, and 321;

[0290] SEQ ID NOs: 26, 44, 150, 235, 309, and 330; SEQ ID NOs: 42, 56, 104, 247, 293, and 385;

[0291] SEQ ID NOs: 27, 77, 172, 262, 278, and 408; SEQ ID NOs: 42, 56, 101, 195, 319, and 379;

[0292] SEQ ID NOs: 26, 44, 161, 239, 310, and 324; SEQ ID NOs: 42, 56, 97, 197, 319, and 375;

[0293] SEQ ID NOs: 43, 80, 149, 243, 317, and 323; SEQ ID NOs: 27, 51, 152, 210, 268, and 357;

[0294] SEQ ID NOs: 25, 78, 119, 231, 293, and 387; SEQ ID NOs: 15, 47, 107, 215, 301, and 341;

[0295] SEQ ID NOs: 42, 56, 102, 258, 274, and 402; SEQ ID NOs: 35, 61, 151, 214, 301, and 347;

[0296] SEQ ID NOs: 42, 56, 95, 239, 310, and 325; SEQ ID NOs: 8, 66, 116, 188, 314, and 340;

[0297] SEQ ID NOs: 14, 85, 153, 220, 299, and 349; SEQ ID NOs: 39, 81, 127, 244, 313, and 335;

[0298] SEQ ID NOs: 42, 57, 132, 191, 305, and 389; SEQ ID NOs: 36, 75, 121, 236, 280, and 337;

[0299] SEQ ID NOs: 25, 77, 168, 221, 301, and 345; SEQ ID NOs: 31, 56, 148, 197, 319, and 378;

[0300] SEQ ID NOs: 42, 56, 106, 256, 285, and 396; SEQ ID NOs: 26, 44, 167, 255, 284, and 398;

[0301] SEQ ID NOs: 42, 56, 95, 190, 310, and 324; SEQ ID NOs: 6, 65, 124, 197, 319, and 373;

[0302] SEQ ID NOs: 17, 62, 120, 196, 319, and 380; SEQ ID NOs: 4, 65, 166, 226, 318, and 336;

[0303] SEQ ID NOs: 42, 57, 96, 239, 310, and 325; SEQ ID NOs: 25, 77, 154, 217, 302, and 343;

[0304] SEQ ID NOs: 42, 56, 115, 263, 281, and 411; SEQ ID NOs: 32, 49, 108, 225, 307, and 410;

[0305] SEQ ID NOs: 42, 56, 95, 242, 308, and 331; SEQ ID NOs: 26, 44, 150, 234, 316, and 326;

[0306] SEQ ID NOs: 42, 56, 95, 192, 311, and 322; SEQ ID NOs: 26, 44, 155, 235, 309, and 330;

[0307] SEQ ID NOs: 25, 77, 168, 213, 301, and 345; SEQ ID NOs: 24, 79, 137, 262, 283, and 406;

[0308] SEQ ID NOs: 42, 56, 100, 248, 307, and 391; SEQ ID NOs: 26, 44, 183, 197, 319, and 378;

[0309] SEQ ID NOs: 25, 77, 93, 251, 293, and 387; SEQ ID NOs: 42, 55, 103, 259, 283, and 400;

[0310] SEQ ID NOs: 42, 56, 92, 262, 278, and 399; SEQ ID NOs: 26, 44, 170, 198, 288, and 352;

[0311] SEQ ID NOs: 25, 77, 165, 251, 291, and 387; SEQ ID NOs: 42, 56, 102, 257, 274, and 402;

[0312] SEQ ID NOs: 25, 77, 111, 253, 292, and 381; SEQ ID NOs: 38, 58, 86, 219, 301, and 348;

[0313] SEQ ID NOs: 42, 56, 122, 262, 283, and 405; SEQ ID NOs: 42, 56, 134, 209, 289, and 364;

[0314] SEQ ID NOs: 32, 50, 162, 216, 300, and 344; SEQ ID NOs: 20, 69, 141, 211, 269, and 356;

[0315] SEQ ID NOs: 12, 73, 140, 199, 297, and 353; SEQ ID NOs: 28, 70, 145, 218, 300, and 390;

[0316] SEQ ID NOs: 21, 74, 135, 210, 267, and 370; SEQ ID NOs: 20, 45, 177, 210, 289, and 363;

[0317] SEQ ID NOs: 28, 46, 184, 204, 287, and 366; SEQ ID NOs: 20, 45, 175, 209, 265, and 350;

[0318] SEQ ID NOs: 26, 74, 178, 209, 286, and 377; SEQ ID NOs: 21, 74, 187, 209, 268, and 374;

[0319] SEQ ID NOs: 19, 44, 142, 200, 297, and 355; SEQ ID NOs: 23, 74, 174, 211, 268, and 351;

[0320] SEQ ID NOs: 10, 74, 89, 209, 289, and 372; SEQ ID NOs: 37, 60, 139, 206, 286, and 339;

[0321] SEQ ID NOs: 41, 83, 157, 232, 306, and 329; SEQ ID NOs: 18, 74, 178, 209, 268, and 372;

[0322] SEQ ID NOs: 32, 49, 179, 224, 300, and 344; SEQ ID NOs: 20, 74, 176, 210, 287, and 359;

[0323] SEQ ID NOs: 26, 74, 178, 209, 287, and 361; SEQ ID NOs: 22, 71, 171, 210, 286, and 360;

[0324] SEQ ID NOs: 11, 44, 159, 208, 286, and 371; SEQ ID NOs: 20, 44, 185, 212, 287, and 365;

[0325] SEQ ID NOs: 41, 84, 136, 238, 271, and 332; SEQ ID NOs: 20, 44, 173, 202, 298, and 367;

[0326] SEQ ID NOs: 20, 74, 182, 209, 286, and 376; SEQ ID NOs: 9, 74, 180, 207, 286, and 362;

[0327] SEQ ID NOs: 41, 82, 158, 240, 270, and 327; or SEQ ID NOs: 29, 44, 181, 209, 286, and 376.

[0328]

[0329] In another specific embodiment, the antibody or antigen-binding fragment thereof is

[0330] It may comprise at least one variable heavy chain region (VH) selected from the group consisting of sequence numbers 412 to 527.

[0331] In another specific embodiment, the antibody or antigen-binding fragment thereof is

[0332] It may comprise at least one variable light chain region (VL) selected from the group consisting of sequence numbers 528 to 643.

[0333]

[0334] In another specific embodiment, the antibody or antigen-binding fragment thereof,

[0335] Containing the VH sequence of SEQ ID NO: 412 and the VL sequence of SEQ ID NO: 528; or

[0336] Containing the VH sequence of SEQ ID NO: 413 and the VL sequence of SEQ ID NO: 529; or

[0337] Containing the VH sequence of SEQ ID NO: 414 and the VL sequence of SEQ ID NO: 530;

[0338] Containing the VH sequence of SEQ ID NO: 415 and the VL sequence of SEQ ID NO: 531;

[0339] Containing the VH sequence of SEQ ID NO: 416 and the VL sequence of SEQ ID NO: 532;

[0340] Containing the VH sequence of SEQ ID NO: 417 and the VL sequence of SEQ ID NO: 533;

[0341] Containing the VH sequence of SEQ ID NO: 418 and the VL sequence of SEQ ID NO: 534;

[0342] Containing the VH sequence of SEQ ID NO: 419 and the VL sequence of SEQ ID NO: 535;

[0343] Containing the VH sequence of SEQ ID NO: 420 and the VL sequence of SEQ ID NO: 536;

[0344] Containing the VH sequence of SEQ ID NO: 421 and the VL sequence of SEQ ID NO: 537;

[0345] Containing the VH sequence of SEQ ID NO: 422 and the VL sequence of SEQ ID NO: 538;

[0346] Containing the VH sequence of SEQ ID NO: 423 and the VL sequence of SEQ ID NO: 539; or

[0347] Containing the VH sequence of SEQ ID NO: 424 and the VL sequence of SEQ ID NO: 540;

[0348] Containing the VH sequence of SEQ ID NO: 425 and the VL sequence of SEQ ID NO: 541; or

[0349] Containing the VH sequence of SEQ ID NO: 426 and the VL sequence of SEQ ID NO: 542;

[0350] Containing the VH sequence of SEQ ID NO: 427 and the VL sequence of SEQ ID NO: 543;

[0351] Containing the VH sequence of SEQ ID NO: 428 and the VL sequence of SEQ ID NO: 544;

[0352] Containing the VH sequence of SEQ ID NO: 429 and the VL sequence of SEQ ID NO: 545; or

[0353] Containing the VH sequence of SEQ ID NO: 430 and the VL sequence of SEQ ID NO: 546;

[0354] Containing the VH sequence of SEQ ID NO: 431 and the VL sequence of SEQ ID NO: 547; or

[0355] Containing the VH sequence of SEQ ID NO: 432 and the VL sequence of SEQ ID NO: 548; or

[0356] Contains the VH sequence of SEQ ID NO: 433 and the VL sequence of SEQ ID NO: 549; or

[0357] It may include the VH sequence of SEQ ID NO: 434 and the VL sequence of SEQ ID NO: 550.

[0358]

[0359] In another specific embodiment, the antibody or antigen-binding fragment thereof,

[0360] Containing the VH sequence of SEQ ID NO: 435 and the VL sequence of SEQ ID NO: 551;

[0361] Containing the VH sequence of SEQ ID NO: 436 and the VL sequence of SEQ ID NO: 552;

[0362] Containing the VH sequence of SEQ ID NO: 437 and the VL sequence of SEQ ID NO: 553;

[0363] Containing the VH sequence of SEQ ID NO: 438 and the VL sequence of SEQ ID NO: 554;

[0364] Containing the VH sequence of SEQ ID NO: 439 and the VL sequence of SEQ ID NO: 555;

[0365] Containing the VH sequence of SEQ ID NO: 440 and the VL sequence of SEQ ID NO: 556;

[0366] Containing the VH sequence of SEQ ID NO: 441 and the VL sequence of SEQ ID NO: 557;

[0367] Containing the VH sequence of SEQ ID NO: 442 and the VL sequence of SEQ ID NO: 558; or

[0368] Containing the VH sequence of SEQ ID NO: 443 and the VL sequence of SEQ ID NO: 559;

[0369] Containing the VH sequence of SEQ ID NO: 444 and the VL sequence of SEQ ID NO: 560;

[0370] Containing the VH sequence of SEQ ID NO: 445 and the VL sequence of SEQ ID NO: 561;

[0371] Containing the VH sequence of SEQ ID NO: 446 and the VL sequence of SEQ ID NO: 562;

[0372] Containing the VH sequence of SEQ ID NO: 447 and the VL sequence of SEQ ID NO: 563;

[0373] Containing the VH sequence of SEQ ID NO: 448 and the VL sequence of SEQ ID NO: 564;

[0374] Containing the VH sequence of SEQ ID NO: 449 and the VL sequence of SEQ ID NO: 565;

[0375] Containing the VH sequence of SEQ ID NO: 450 and the VL sequence of SEQ ID NO: 566;

[0376] Containing the VH sequence of SEQ ID NO: 451 and the VL sequence of SEQ ID NO: 567;

[0377] Containing the VH sequence of SEQ ID NO: 452 and the VL sequence of SEQ ID NO: 568;

[0378] Containing the VH sequence of SEQ ID NO: 453 and the VL sequence of SEQ ID NO: 569; or

[0379] Containing the VH sequence of SEQ ID NO: 454 and the VL sequence of SEQ ID NO: 570;

[0380] Containing the VH sequence of SEQ ID NO: 455 and the VL sequence of SEQ ID NO: 571;

[0381] Containing the VH sequence of SEQ ID NO: 456 and the VL sequence of SEQ ID NO: 572;

[0382] Containing the VH sequence of SEQ ID NO: 457 and the VL sequence of SEQ ID NO: 573;

[0383] Containing the VH sequence of SEQ ID NO: 458 and the VL sequence of SEQ ID NO: 574; or

[0384] Containing the VH sequence of SEQ ID NO: 459 and the VL sequence of SEQ ID NO: 575;

[0385] Containing the VH sequence of SEQ ID NO: 460 and the VL sequence of SEQ ID NO: 576;

[0386] Containing the VH sequence of SEQ ID NO: 461 and the VL sequence of SEQ ID NO: 577;

[0387] Containing the VH sequence of SEQ ID NO: 462 and the VL sequence of SEQ ID NO: 578;

[0388] Containing the VH sequence of SEQ ID NO: 463 and the VL sequence of SEQ ID NO: 579; or

[0389] Containing the VH sequence of SEQ ID NO: 464 and the VL sequence of SEQ ID NO: 580;

[0390] Containing the VH sequence of SEQ ID NO: 465 and the VL sequence of SEQ ID NO: 581;

[0391] Containing the VH sequence of SEQ ID NO: 466 and the VL sequence of SEQ ID NO: 582;

[0392] Containing the VH sequence of SEQ ID NO: 467 and the VL sequence of SEQ ID NO: 583;

[0393] Containing the VH sequence of SEQ ID NO: 468 and the VL sequence of SEQ ID NO: 584;

[0394] Containing the VH sequence of SEQ ID NO: 469 and the VL sequence of SEQ ID NO: 585;

[0395] Containing the VH sequence of SEQ ID NO: 470 and the VL sequence of SEQ ID NO: 586;

[0396] Containing the VH sequence of SEQ ID NO: 471 and the VL sequence of SEQ ID NO: 587;

[0397] Containing the VH sequence of SEQ ID NO: 472 and the VL sequence of SEQ ID NO: 588; or

[0398] Containing the VH sequence of SEQ ID NO: 473 and the VL sequence of SEQ ID NO: 589; or

[0399] Containing the VH sequence of SEQ ID NO: 474 and the VL sequence of SEQ ID NO: 590;

[0400] Containing the VH sequence of SEQ ID NO: 475 and the VL sequence of SEQ ID NO: 591; or

[0401] Containing the VH sequence of SEQ ID NO: 476 and the VL sequence of SEQ ID NO: 592;

[0402] Containing the VH sequence of SEQ ID NO: 477 and the VL sequence of SEQ ID NO: 593;

[0403] Containing the VH sequence of SEQ ID NO: 478 and the VL sequence of SEQ ID NO: 594;

[0404] Containing the VH sequence of SEQ ID NO: 479 and the VL sequence of SEQ ID NO: 595; or

[0405] Containing the VH sequence of SEQ ID NO: 480 and the VL sequence of SEQ ID NO: 596;

[0406] Containing the VH sequence of SEQ ID NO: 481 and the VL sequence of SEQ ID NO: 597; or

[0407] Containing the VH sequence of SEQ ID NO: 482 and the VL sequence of SEQ ID NO: 598; or

[0408] Containing the VH sequence of SEQ ID NO: 483 and the VL sequence of SEQ ID NO: 599;

[0409] Containing the VH sequence of SEQ ID NO: 484 and the VL sequence of SEQ ID NO: 600;

[0410] Containing the VH sequence of SEQ ID NO: 485 and the VL sequence of SEQ ID NO: 601;

[0411] Containing the VH sequence of SEQ ID NO: 486 and the VL sequence of SEQ ID NO: 602;

[0412] Containing the VH sequence of SEQ ID NO: 487 and the VL sequence of SEQ ID NO: 603;

[0413] Containing the VH sequence of SEQ ID NO: 488 and the VL sequence of SEQ ID NO: 604;

[0414] Containing the VH sequence of SEQ ID NO: 489 and the VL sequence of SEQ ID NO: 605;

[0415] Containing the VH sequence of SEQ ID NO: 490 and the VL sequence of SEQ ID NO: 606;

[0416] Containing the VH sequence of SEQ ID NO: 491 and the VL sequence of SEQ ID NO: 607; or

[0417] Containing the VH sequence of SEQ ID NO: 492 and the VL sequence of SEQ ID NO: 608;

[0418] Containing the VH sequence of SEQ ID NO: 493 and the VL sequence of SEQ ID NO: 609;

[0419] Containing the VH sequence of SEQ ID NO: 494 and the VL sequence of SEQ ID NO: 610;

[0420] Containing the VH sequence of SEQ ID NO: 495 and the VL sequence of SEQ ID NO: 611;

[0421] Containing the VH sequence of SEQ ID NO: 496 and the VL sequence of SEQ ID NO: 612;

[0422] Containing the VH sequence of SEQ ID NO: 497 and the VL sequence of SEQ ID NO: 613;

[0423] Containing the VH sequence of SEQ ID NO: 498 and the VL sequence of SEQ ID NO: 614;

[0424] Containing the VH sequence of SEQ ID NO: 499 and the VL sequence of SEQ ID NO: 615;

[0425] Containing the VH sequence of SEQ ID NO: 500 and the VL sequence of SEQ ID NO: 616;

[0426] Containing the VH sequence of SEQ ID NO: 501 and the VL sequence of SEQ ID NO: 617; or

[0427] Containing the VH sequence of SEQ ID NO: 502 and the VL sequence of SEQ ID NO: 618; or

[0428] Containing the VH sequence of SEQ ID NO: 503 and the VL sequence of SEQ ID NO: 619; or

[0429] Containing the VH sequence of SEQ ID NO: 504 and the VL sequence of SEQ ID NO: 620;

[0430] Containing the VH sequence of SEQ ID NO: 505 and the VL sequence of SEQ ID NO: 621; or

[0431] Containing the VH sequence of SEQ ID NO: 506 and the VL sequence of SEQ ID NO: 622;

[0432] Containing the VH sequence of SEQ ID NO: 507 and the VL sequence of SEQ ID NO: 623;

[0433] Containing the VH sequence of SEQ ID NO: 508 and the VL sequence of SEQ ID NO: 624;

[0434] Containing the VH sequence of SEQ ID NO: 509 and the VL sequence of SEQ ID NO: 625;

[0435] Containing the VH sequence of SEQ ID NO: 510 and the VL sequence of SEQ ID NO: 626;

[0436] Containing the VH sequence of SEQ ID NO: 511 and the VL sequence of SEQ ID NO: 627;

[0437] Containing the VH sequence of SEQ ID NO: 512 and the VL sequence of SEQ ID NO: 628;

[0438] Containing the VH sequence of SEQ ID NO: 513 and the VL sequence of SEQ ID NO: 629;

[0439] Containing the VH sequence of SEQ ID NO: 514 and the VL sequence of SEQ ID NO: 630;

[0440] Containing the VH sequence of SEQ ID NO: 515 and the VL sequence of SEQ ID NO: 631;

[0441] Containing the VH sequence of SEQ ID NO: 516 and the VL sequence of SEQ ID NO: 632;

[0442] Containing the VH sequence of SEQ ID NO: 517 and the VL sequence of SEQ ID NO: 633;

[0443] Containing the VH sequence of SEQ ID NO: 518 and the VL sequence of SEQ ID NO: 634;

[0444] Containing the VH sequence of SEQ ID NO: 519 and the VL sequence of SEQ ID NO: 635; or

[0445] Containing the VH sequence of SEQ ID NO: 520 and the VL sequence of SEQ ID NO: 636;

[0446] Containing the VH sequence of SEQ ID NO: 521 and the VL sequence of SEQ ID NO: 637;

[0447] Containing the VH sequence of SEQ ID NO: 522 and the VL sequence of SEQ ID NO: 638;

[0448] Containing the VH sequence of SEQ ID NO: 523 and the VL sequence of SEQ ID NO: 639;

[0449] Containing the VH sequence of SEQ ID NO: 524 and the VL sequence of SEQ ID NO: 640;

[0450] Containing the VH sequence of SEQ ID NO: 525 and the VL sequence of SEQ ID NO: 641;

[0451] Contains the VH sequence of SEQ ID NO: 526 and the VL sequence of SEQ ID NO: 642; or

[0452] It may include the VH sequence of SEQ ID NO: 527 and the VL sequence of SEQ ID NO: 643.

[0453] In one specific example, the antibody provided in the present invention may be a monoclonal antibody.

[0454] In one specific example, the antigen-binding fragment provided in the present invention may be scFv.

[0455]

[0456] The amino acid sequences described above may include variants having different sequences due to deletion, insertion, substitution, or a combination thereof of amino acid residues, as long as the structure, function, activity, etc. of the polypeptide including the amino acid sequences are not affected. In addition, the amino acid sequences may include amino acids that have undergone conventional modifications known in the art, and the amino acid modifications may be, for example, phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, etc.

[0457] The antibody or antigen-binding fragment thereof of the present invention includes not only the amino acid sequences described above, but also those having substantially the same amino acid sequence as the amino acid sequence described above or variants thereof. The meaning of having the substantially same amino acid sequence may include an amino acid sequence having 90% or more, 91% or more, 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more, or 99.5% or more homology to the amino acid sequence described above, but is not limited thereto.

[0458] The antibody or antigen-binding fragment of the present invention may further comprise a linker between the heavy chain variable region and the light chain variable region. The linker may be a peptide linker and have a length of about 10-25 amino acids. For example, the linker may include, but is not limited to, hydrophilic amino acids such as glycine and / or serine.

[0459] Specifically, in the present invention, the linker may include, for example, (GS)n, (GGS)n, (GSGGS)n or (GnS)m (n and m are each 1 to 10), and preferably may include GGGGS.

[0460] The antibody or antigen-binding fragment of the present invention may include not only the sequence of the anti-MYCT1 antibody or antigen-binding fragment of the present invention described herein, but also biological equivalents thereof, as long as it can specifically recognize MYCT1. For example, additional changes may be made to the amino acid sequence of the antibody to further improve the binding affinity and / or other biological properties of the antibody. Such modifications include, for example, deletions, insertions, and / or substitutions of amino acid sequence residues of the antibody or antigen-binding fragment. Such amino acid mutations are made based on the relative similarity of amino acid side-chain substituents, such as hydrophobicity, hydrophilicity, charge, size, etc. Analysis of the size, shape, and type of amino acid side-chain substituents reveals that arginine, lysine, and histidine are all positively charged residues; alanine, glycine, and serine have similar sizes; and phenylalanine, tryptophan, and tyrosine have similar shapes. Therefore, based on these considerations, arginine, lysine and histidine; alanine, glycine and serine; and phenylalanine, tryptophan and tyrosine can be considered biologically functional equivalents.

[0461] Considering the mutations having the above-described biological equivalent activity, the antibody or antigen-binding fragment of the present invention or the nucleic acid molecule encoding them is interpreted to also include a sequence showing substantial identity with the sequence described in the sequence number. The substantial identity means a sequence showing at least 90% homology, most preferably at least 95% homology, 96% or more, 97% or more, 98% or more, or 99% or more homology when the sequence of the present invention is aligned with any other sequence to the greatest extent possible and the aligned sequence is analyzed using an algorithm commonly used in the art. Alignment methods for sequence comparison are known in the art. NCBI Basic Local Alignment Search Tool (BLAST) is accessible from NBCI, etc., and can be used in conjunction with sequence analysis programs such as blastp, blasm, blastx, tblastn, and tblastx on the Internet. BLAST is available at www.ncbi.nlm.nih.gov / BLAST / . Instructions for comparing sequence homology using this program can be found at www.ncbi.nlm.nih.gov / BLAST / blast_help.html.

[0462] Based on this, the antibody or antigen-binding fragment of the present invention may have 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more homology to the specified sequences or the entirety of the sequences described herein. Such homology may be determined by sequence comparison and / or alignment using methods known in the art. For example, the percent sequence homology of the nucleic acids or proteins of the present invention may be determined using sequence comparison algorithms (e.g., BLAST or BLAST 2.0), manual alignment, or visual inspection.

[0463] The binding affinity of the anti-MYCT1 human antibody or antigen-binding fragment thereof provided in the present invention to MYCT1 is 10 -5 M to 10 -12 is within the M range. For example, the binding affinity of an anti-MYCT1 human antibody or antigen-binding fragment to MYCT1 is 10 -6 M to 10 -12 M, 10 -7 M to 10 -12 M, 10 -8 M to 10 -12 M, 10 -9 M to 10 -12 M, 10 -5 M to 10 -11 M, 10 -6 M to 10 -11 M, 10 -7 M to 10 -11 M, 10 -8 M to 10 -11 M, 10 -9 M to 10 -11 M, 10 -10 M to 10 -11 M, 10 -5 M to 10 -10 M, 10 -6 M to 10 -10 M, 10 -7 M to 10 -10 M, 10 -8 M to 10 -10 M, 10 -9 M to 10 -10 M, 10 -5 M to 10 -9 M, 10 -6 M to 10 -9 M,10 -7 M to 10 -9 M, 10 -8 M to 10 -9 M, 10 -5 M to 10 -8 M, 10 -6 M to 10 -8 M, 10 -7 M to 10 -8 M, 10-5 M to 10 -7 M, 10 -6 M to 10 -7 M or 10 -5 M to 10 -6 M may be, but is not limited to, this.

[0464] According to another embodiment of the present invention, the present invention relates to a nucleic acid molecule encoding an antibody or antigen-binding fragment provided by the present invention.

[0465] The term "nucleic acid" as used herein encompasses DNA (gDNA and cDNA) and RNA molecules, and nucleotides, the basic structural units of nucleic acids, include not only natural nucleotides but also analogues with modified sugar or base moieties. The sequence of the nucleic acid encoding the heavy and light chain variable regions of the present invention may be modified. Such modifications include additions, deletions, or non-conservative or conservative substitutions of nucleotides.

[0466] In the present invention, the DNA encoding the antibody or antigen-binding fragment can be easily isolated or synthesized using conventional molecular biological techniques (for example, by using an oligonucleotide probe that can specifically bind to DNA encoding the antibody and the heavy and light chains), and the nucleic acid is isolated and inserted into a replicable vector for further cloning (amplification of DNA) or further expression.

[0467]

[0468] According to another embodiment, the present invention relates to a recombinant expression vector comprising a nucleic acid molecule provided in the present invention.

[0469] In the present invention, the above "vector" refers to a recombinant vector that can express a target protein when transfected into a suitable host cell, and a genetic construct that includes essential regulatory elements operably linked to allow the gene insert to be expressed. Here, the term "operably linked" means that a nucleic acid expression regulatory sequence and a nucleic acid sequence encoding a target protein are functionally linked to perform a general function. The operably linked vector can be produced using a genetic recombination technique well known in the art, and site-specific DNA cleavage and ligation can be easily performed using enzymes generally known in the art.

[0470] In the present invention, various types of vectors such as nanoparticles, plasmids, viruses, and cosmids can be used as recombinant expression vectors for inserting the foreign gene. The type of recombinant vector is not particularly limited as long as it has the function of expressing a desired gene and producing a desired protein in various host cells of prokaryotic and eukaryotic cells. Specifically, a vector that has a promoter exhibiting strong activity and a strong expression ability while being capable of mass-producing a foreign protein in a form similar to that in the natural state can be used.

[0471] A variety of gene delivery vehicles are known in the art, including both viral and non-viral (e.g., naked DNA, plasmid) vectors. Viral vectors suitable for gene delivery are known to those skilled in the art. Non-limiting examples of the viral vectors may include retroviral vectors (e.g., derived from Moloney murine leukemia virus vector (MoMLV), MSCV, SFFV, MPSV, SNV, etc.), lentiviral vectors (e.g., derived from HIV-1, HIV-2, SIV, BIV, FIV, etc.), adenovirus (Ad) vectors including replication competent, replication deficient and emasculated forms thereof, adeno-associated virus (AAV) vectors, simian virus 40 (SV-40) vectors, bovine papilloma virus vectors, Epstein-Barr virus vectors, herpes virus vectors, varicella virus vectors, Harvey murine sarcoma virus vectors, murine mammary tumor virus vectors, Rous sarcoma virus vectors, parvovirus vectors, poliovirus vectors, vesicular stomatitis virus vectors, Maraba virus vectors and group B adenovirus enadenotuxireb vectors.

[0472] Non-viral vectors for gene transfer include naked DNA, plasmids, transposons, and mRNA. Non-limiting examples include pKK plasmid (Clonetech), pUC plasmid, pET plasmid (Novagen, Inc., Madison, Wis.), pRSET or pREP plasmids (Invitrogen, San Diego, Calif.), and pMAL plasmid (New England Biolabs, Beverly, Mass.).

[0473] The vector of the present invention can be introduced into many suitable host cells using methods disclosed or cited herein or otherwise known to those skilled in the art.

[0474] Suitable expression vectors of the present invention may include, in addition to expression regulatory elements such as a promoter, initiation codon, termination codon, polyadenylation signal, or enhancer, a nucleotide sequence encoding a signal peptide for membrane targeting or secretion. The initiation and termination codons are generally considered to be part of the nucleotide sequence encoding the immunogenic target protein, and must be functional in a subject when the genetic construct is administered, and must be in frame with the coding sequence.

[0475] The term "promoter," as used herein, refers to any sequence that regulates the expression of a coding sequence, such as a gene. A promoter may be, for example, constitutive, inducible, repressible, or tissue-specific. A promoter is a control sequence, a region of a polynucleotide sequence where the initiation and rate of transcription are controlled. Non-limiting examples of the promoter in the present invention include Rous sarcoma virus (RSV) LTR promoter (optionally with RSV enhancer), cytomegalovirus (CMV) promoter, SV40 promoter, dihydrofolate reductase promoter, β-actin promoter, phosphoglycerol kinase (PGK) promoter, U6 promoter, EF1alpha short form (EFS) promoter, human polypeptide chain elongation factor (EF1a) promoter, P5 promoter, Ubc promoter, CAG promoter, TRE promoter, UAS promoter, Ac5 promoter, polyhedrin promoter, CaMKIIa promoter, Gal1 promoter, TEF1 promoter, GDS promoter, ADH1 promoter, CaMV35S promoter, ubiquitin (Ubi) Promoters such as ubiquitin C (UbiC), H1 promoter, U6 promoter, alpha-1-antitrypsin promoter, spleen focus-forming virus (SFFV) promoter, etc. may be included.

[0476] Additionally, in the present invention, the promoter may be coupled to an enhancer to increase transcription efficiency. Non-limiting examples of the enhancer include, but are not limited to, an RSV enhancer, a CMV enhancer, or an α-fetoprotein MERII enhancer. The vector of the present invention may also be fused with other sequences to facilitate the purification of antibodies expressed therefrom. Examples of the fused sequences include, but are not limited to, glutathione S-transferase (Pharmacia, USA), maltose binding protein (NEB, USA), FLAG (IBI, USA), and 6x His (hexahistidine; Quiagen, USA).

[0477] The vector of the present invention includes an antibiotic resistance gene commonly used in this technical field as a selectable marker, and may include, but is not limited to, resistance genes for ampicillin, gentamicin, carbenicillin, chloramphenicol, streptomycin, kanamycin, geneticin, neomycin, and tetracycline.

[0478]

[0479] According to another embodiment of the present invention, the expression vector provided in the present invention relates to a transfected host cell.

[0480] The term "host cell" as used herein includes an individual cell or cell culture that can be or has been the recipient of a vector(s) for incorporation of a polypeptide insert. Host cells include progeny of a single host cell, which progeny may not necessarily be completely identical (morphologically or in genomic DNA complement) to the original parent cell due to natural, accidental, or intentional mutation. Host cells include cells that have been transfected in vivo with the polypeptide(s) of the present disclosure.

[0481] In the present invention, the host cell may include a mammalian, plant, insect, fungal or cellular cell, and for example, bacterial cells such as Escherichia coli, Streptomyces, and Salmonella Typhimurium; fungal cells such as yeast cells and Pichia pastoris; insect cells such as Drozophylla and Spodoptera Sf9 cells; animal cells such as CHO (Chinese hamster ovary cells), SP2 / 0 (mouse myeloma), human lymphoblastoid, COS, NSO (mouse myeloma), 293T, Bow melanoma cells, HT-1080, BHK (Baby Hamster Kidney cells), HEK (Human Embryonic Kidney cells), or PERC.6 (human retinal cells); Alternatively, the host cell may be a plant cell, but is not limited thereto, and any cell known to a person skilled in the art that can be used as a host cell line may be used.

[0482] In the present invention, the transfection method is any method for injecting the desired vector into the host cell, and any known method capable of injecting a vector into a host cell can be included, and for example, a method using CaCl2, electroporation, microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, DEAE-dextran treatment, gene bombardment, and virus-based transfection can be used, but is not limited thereto.

[0483] According to another embodiment, the present invention relates to a method for producing an antibody or antigen-binding fragment that specifically binds to MYCT1, comprising the steps of: culturing the host cell to produce an antibody or antigen-binding fragment; and isolating and purifying the produced antibody or antigen-binding fragment.

[0484] In the present invention, the host cells can be cultured in various media. Any commercially available media can be used as culture media. Any other essential supplements known to those skilled in the art may be included at appropriate concentrations. Culture conditions, such as temperature, pH, etc., are already used with host cells selected for expression and will be readily apparent to those skilled in the art.

[0485] In the present invention, the antibody or antigen-binding fragment thereof can be recovered by removing impurities, for example, by centrifugation or ultrafiltration, and purifying the resultant product using, for example, affinity chromatography. Additional purification techniques, for example, anion or cation exchange chromatography, hydrophobic interaction chromatography, hydroxylapatite chromatography, etc., can be used.

[0486]

[0487] According to another embodiment of the present invention, there is provided a dual or multispecific antibody comprising an antibody or antigen-binding fragment provided by the present invention.

[0488] The term "bispecific antibody" as used herein means an antibody having binding ability or antagonism to more than one target, and means an antibody in which antibodies having binding ability or antagonism to two different targets are combined, or an antibody in which an antibody having binding ability to one target and a substance having antagonism to another target are combined.

[0489] The term "multispecific antibody" as used herein refers to an antibody having binding specificities for at least three different antigens. Multispecific antibodies may include antibodies that target more than three specific targets, such as trispecific antibodies, tetraspecific antibodies, or antibodies that target more than one target.

[0490] In the present invention, antibodies belonging to bispecific or multispecific antibodies can be classified into scFv-based antibodies, Fab-based antibodies, and IgG-based antibodies. In the case of bispecific or multispecific antibodies, since two or more signals can be simultaneously suppressed or amplified, they can be more effective than cases where a single signal is suppressed / amplified. In addition, compared to cases where each signal is treated with a separate signal inhibitor, lower dosage administration is possible, and two or more signals can be suppressed / amplified in the same time and space.

[0491] Methods for producing bispecific or multispecific antibodies are well known. Traditionally, recombinant production of bispecific antibodies is based on the co-expression of two or more immunoglobulin heavy chain / light chain pairs, where the two or more heavy chains have different specificities.

[0492] In the present invention, in the case of a bispecific or multispecific antibody based on scFv, the VL and VH of different scFvs can be combined with each other to produce a hybrid scFv in a heterodimeric form to produce a diabody, different scFvs can be linked to each other to produce a tendem ScFv, CH1 and CL of Fab can be expressed at the ends of each scFv to produce a heterodimeric miniantibody, and some amino acids of the CH3 domain, which is a homodimeric domain of Fc, can be substituted to change it into a heterodimeric structure in the form of a 'knob into hole', and these changed CH3 domains can be expressed at the ends of different scFvs to produce a minibody in the form of a heterodimeric scFv.

[0493] In the present invention, in the case of a bispecific or multispecific antibody based on Fab, individual Fabs for specific antigens can be combined with each other using a disulfide bond or a mediator to produce a heterodimeric Fab form, and by expressing scFvs for different antigens at the terminals of the heavy or light chains of a specific Fab, it can be produced to have two antigen-binding valencies, or by providing a hinge region between the Fab and scFv, it can be produced to have four antigen-binding valencies in a homodimeric form. In addition, a dual-target bibody in which the antigen-binding valencies are made three by fusing scFvs for different antigens to the light and heavy chain terminals of the Fab, and a tri-target bibody in which the antigen-binding valencies are made three by fusing different scFvs to the light and heavy chain terminals of the Fab, respectively, can be obtained by chemically conjugating three different Fabs.

[0494] In the case of IgG-based bispecific or multispecific antibodies in the present invention, a method for producing bispecific antibodies by producing hybrid hybridomas, also known as quadromas, by crossbreeding mouse and rat hybridomas again has been known by Trion Pharma. In addition, bispecific antibodies can be produced in the so-called 'Holes and Knob' form, which is produced in a heterodimeric form by modifying some amino acids in the CH3 homodimeric domain of Fc for different heavy chains while sharing the light chain portion. In addition to the heterodimeric form of bispecific antibodies, two different scFvs can be fused and expressed in the constant domains of the light and heavy chains of IgG instead of the variable domains, thereby producing (scFv)4-IgG in a homodimeric form. In addition, ImClone reported the production of a bispecific antibody by fusing only a single variable domain for mouse platelet-derived growth factor receptor-α to the light chain amino terminus of IMC-1C11, a chimeric monoclonal antibody against human VEGFR-2. In addition, antibodies with multiple antigen-binding affinities for CD20 can be produced through the so-called 'dock and lock (DNL)' method using the dimerization and docking domain (DDD) of the R subunit of protein kinase A (PKA) and the anchoring domain of PKA.

[0495] In the present invention, a wide variety of recombinant antibody formats have been developed, for example, bivalent, trivalent, or tetravalent bispecific or multispecific antibodies. For example, it also includes bivalent, trivalent, or tetravalent antibodies described in International Patent Application Publication Nos. WO2001 / 077342, WO2009 / 080251, WO2009 / 080252, WO2009 / 080253, WO2009 / 080254, WO2010 / 112193, WO2010 / 115589, WO2010 / 136172, WO2010 / 145792, WO2010 / 145793, and WO2011 / 117330. An antibody being bivalent, trivalent, or tetravalent indicates that two or more binding domains, three or more binding domains, or four or more binding domains are present in the antibody molecule, respectively.

[0496] The bispecific or multispecific antibody according to the present invention may specifically comprise the anti-MYCT1 antibody or antigen-binding fragment in the form of an IgG complete antibody or a fragment thereof, for example, a single-chain Fv, a VH domain and / or a VL domain, a Fab or (Fab)2. In addition, the antibody targeting MYCT1 may comprise an antibody that binds to a target other than the antibody targeting MYCT1, for example, PD-1, PD-L1, BTLA, CTLA-4, VISTA, LAG3, TIM3, CD137 (4-1BB), VISTA, CD258 (LIGHT), TIGIT, CD134 (OX40), CD28, CD278 (ICOS), CD27, CD154 (CD40L), CD357 (GITR), CD30, DR3, CD226 (DNAM1), CD96, CD200, CD200R, Transferrin receptor, c-Met, EGFR, HER2, KDR, PDGFRa, An antibody targeting one or more selected from the group consisting of NRP1, MARCO, specifically an IgG complete antibody or fragment thereof, for example in the form of a single chain Fv, VH domain and / or VL domain, Fab or (Fab)2.

[0497] The bispecific or multispecific antibodies according to the present invention may provide additional binding specificities induced or mediated by targets other than MYCT1.

[0498] For example, the bispecific antibody according to the present invention binds to MYCT1 and PD-1, PD-L1, BTLA, CTLA-4, VISTA, LAG3, TIM3, CD137 (4-1BB), VISTA, CD258 (LIGHT), TIGIT, CD3, CD20, CD134 (OX40), CD28, CD278 (ICOS), CD27, CD154 (CD40L), CD357 (GITR), CD30, DR3, CD226 (DNAM1), CD96, CD200, CD200R, Transferrin receptor, c-Met, EGFR, HER2, KDR, PDGFRa, NRP1, MARCO, FcRH5, HER2, LYPD1, LY6G6D, PMEL17, LY6E, NKG2D, CD3, CD16a, CD19, CD33, CD22, It can simultaneously target one or more selected from the group consisting of CD79A, CD79B, EDAR, GFRA1, MRP4, RET, Steap1, and TenB2.

[0499] For example, the multispecific antibody according to the present invention may bind to MYCT1 and PD-1, PD-L1, BTLA, CTLA-4, VISTA, LAG3, TIM3, CD137 (4-1BB), VISTA, CD258 (LIGHT), TIGIT, CD3, CD20, CD134 (OX40), CD28, CD278 (ICOS), CD27, CD154 (CD40L), CD357 (GITR), CD30, DR3, CD226 (DNAM1), CD96, CD200, CD200R, Transferrin receptor, c-Met, EGFR, HER2, KDR, PDGFRa, NRP1, MARCO, FcRH5, HER2, LYPD1, LY6G6D, PMEL17, LY6E, NKG2D, CD3, CD16a, CD19, CD33, CD22, It can simultaneously target two or more selected from the group consisting of CD79A, CD79B, EDAR, GFRA1, MRP4, RET, Steap1, and TenB2.

[0500] According to another embodiment of the present invention, there is provided an immune cell engaging bispecific or multispecific antibody comprising an antigen binding fragment (scFv) provided by the present invention and at least one scFv that binds to an immune cell activating antigen.

[0501] In the present invention, a cytolytic synapse between cytotoxic T cells or NK cells and cancer target cells is temporarily induced through the immune cell engaging bispecific or multispecific antibody, thereby causing the release of toxic substances.

[0502] In the present invention, the T cell activation antigen may be, but is not limited to, CD3, TCRα, TCRβ, TCRγ, TCRξ, HVEM, LIGHT, CD40, 4-1BB, OX40, DR3, GITR, CD30, TIM1, SLAM, CD2 or CD226.

[0503] In the present invention, the NK cell activating antigen may be, but is not limited to, NKp30, NKp40, NKp44, NKp46, NKG2D, DNAM1, DAP10, CD16 (eg, CD16a, CD16b), CRTAM, CD27, PSGL1, CD96, CD100 (SEMA4D), NKp80, CD244 (SLAMF4 or 2B4), SLAMF6, SLAMF7, KIR2DS2, KIR2DS4, KIR3DS1, KIR2DS3, KIR2DS5, KIR2DS1, CD94, NKG2C, NKG2E or CD160.

[0504] Other immune cell engagers are specifically described in U.S. Patent Application Publication No. US 2017 / 0368169, which is incorporated herein by reference.

[0505] Specifically, the immune cell engaging bispecific or multispecific antibody comprises a tandem scFv and can bind to the following antigen and a surface antigen on a cancer cell. The surface antigen on the cancer cell is MYCT1, which is targeted by the antibody according to the present invention.

[0506] In the present invention, the immune cell engaging bispecific or multispecific antibody may include a structure in the form of, for example, VL(MYCT1)-VH(MYCT1)-VH(CD3 or CD16A)-VL(CD3 or CD16A), VH(MYCT1)-VL(MYCT1)-VH(CD3 or CD16A)-VL(CD3 or CD16A), VH(CD3 or CD16A)-VL(CD3 or CD16A)-VH(MYCT1)-VL(MYCT1) or VH(CD3 or CD16A)-VL(CD3 or CD16A)-VL(MYCT1)-VH(MYCT1).

[0507] The present invention may further include a linker between the heavy chain variable region and the light chain variable region. The linker may be a peptide linker and have a length of about 10-25 amino acids. For example, the linker may include, but is not limited to, hydrophilic amino acids such as glycine and / or serine.

[0508] Specifically, in the present invention, the linker may include, for example, (GS)n, (GGS)n, (GSGGS)n or (GnS)m (n and m are each 1 to 10), preferably GGGGS, and preferably GGGGSGGGGSGGGGS, but is not limited thereto.

[0509] Examples of the immune cell-engaging bispecific or multispecific antibodies in the present invention include blinatumomab (Amgen), which binds to CD3 and CD19; solitomab (Amgen), which binds to CD3 and EpCAM; MEDI 565 (MedImmune, Amgen), which binds to CD3 and CEA; and BAY2010112 (Bayer, Amgen), which binds to CD3 and PSMA. Exemplary DARTs include MGD006 (Macrogenics), which binds to CD3 and CD123; and MGD007 (Macrogenics), which binds to CD3 and gpA33. Exemplary TandAbs may include AFM11 (Affimed Therapeutics), which binds to CD3 and CD19; and AFM13 (Affimed Therapeutics), which binds to CD30 and CD16A.

[0510]

[0511] immunoconjugate

[0512] According to another embodiment of the present invention, the present invention relates to an immunoconjugate in which an antibody or antigen-binding fragment provided by the present invention is combined with a cytotoxic agent.

[0513] In the present invention, the cytotoxic agent may be at least one selected from the group consisting of a chemotherapeutic agent or a chemotherapeutic drug, a growth inhibitor, a toxin (e.g., a protein toxin, an enzymatically active toxin or fragment thereof derived from bacteria, fungi, plants or animals), and a radioactive isotope.

[0514] As an example of implementation, the immune conjugate provided by the present invention may be an antibody-drug conjugate (ADC) in which the antibody or antigen-binding fragment is bound to a drug.

[0515] The term "antibody-drug conjugate (ADC)" used herein refers to a form in which a drug and an antibody are chemically linked without reducing the biological activity of the antibody and the drug. In the present invention, the antibody-drug conjugate refers to a form in which a drug is bound to an amino acid residue at the N-terminus of the heavy chain and / or light chain of an antibody, specifically, a form in which a drug is bound to an α-amine group at the N-terminus of the heavy chain and / or light chain of an antibody.

[0516] In the present invention, the antibody or antigen-binding fragment may be bound to a drug via a linker. The linker serves as a linking moiety between the antibody and the drug. It must be cleavable under intracellular conditions, i.e., enable the drug to be released from the antibody in the intracellular environment. Reflecting the antibody's long half-life, the antibody must be stable in systemic circulation, and the linker-drug binding must not affect the stability or pharmacokinetics of the antibody.

[0517] In the present invention, the linker may include, for example, a cleavable linker or a non-cleavable linker. In the case of a cleavable linker, like a peptide linker, it can be cleaved by an intracellular peptidase or protease enzyme, such as a lysosomal or endosomal protease, and in the case of a non-cleavable linker, for example, a thioether linker, the drug can be released after the antibody is non-selectively degraded by intracellular hydrolysis.

[0518] In the present invention, the cleavable linker may include a peptide linker. The peptide linker has a length of at least two amino acids. For example, it may include a Val-Cit, Val-Ala, or Val-Cit dipeptide, or Phe-Leu or Gly-Phe-Leu-Gly. Examples of linkers are specifically described in International Patent Application Publication No. WO2004 / 010957, which may be incorporated herein by reference.

[0519] In the present invention, the antibody-drug conjugate binds to an antigen on a target cancer cell, forming an ADC-antigen complex, which is then internalized into the cancer cell via the endosome-lysosome pathway. In this case, the intracellular release of the cytotoxic drug is controlled by the internal environment of the endosome / lysosome.

[0520] In the present invention, the cleavable linker is pH-sensitive and may be susceptible to hydrolysis at a specific pH value. Generally, a pH-sensitive linker indicates that it can be hydrolyzed under acidic conditions. For example, it may be an acid-labile linker that can be hydrolyzed in lysosomes, such as a hydrazone, a semicarbazone, a thiosemicarbazone, a cis-aconitic amide, an orthoester, an acetal, a ketal, etc.

[0521] In the present invention, the linker may be cleaved under reducing conditions, and for example, a disulfide linker may correspond thereto. Various disulfide bonds may be formed using SATA (N-succinimidyl-S-acetylthioacetate), SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate) and SMPT (N-succinimidyl-oxycarbonyl-alpha-methyl-alpha-(2-pyridyl-dithio)toluene). These disulfide linkers can be cleaved by disulfide exchange with the thiol of intracellular glutathione.

[0522] In the present invention, the drug and / or drug-linker may be randomly conjugated via lysine of the antibody, or conjugated via cysteine ​​exposed when the disulfide bond chain is reduced. In some cases, the linker-drug may be conjugated via cysteine ​​present in a genetically engineered tag, for example, a peptide or a protein. The genetically engineered tag, for example, a peptide or a protein, may include an amino acid motif that can be recognized by, for example, an isoprenoid transferase. The peptide or protein has a deletion at the carboxyl terminus of the peptide or protein, or has an addition via covalent bonding of a spacer unit to the carboxyl (C) terminus of the peptide or protein.

[0523] In the present invention, the peptide or protein may be directly covalently linked to the amino acid motif or may be covalently linked to a spacer unit to be linked to the amino acid motif. The amino acid spacer unit is composed of 1 to 20 amino acids, and among them, a glycine unit is preferred.

[0524] In the present invention, the linker may include a beta-glucuronide linker that is recognized and hydrolyzed by beta-glucuronidase, which exists in large numbers in lysosomes or is overexpressed in some tumor cells. Unlike a peptide linker, it has a high hydrophilicity, and thus has an advantage of increasing the solubility of an antibody-drug complex when bound to a drug with a high hydrophobic property. In this regard, a beta-glucuronide linker disclosed in International Patent Application Publication No. WO2015 / 182984, for example, a beta-glucuronide linker including a self-immolative group, may be used, and the above document is incorporated by reference.

[0525] In the present invention, the linker may be, for example, a non-cleavable linker, and the drug is released through only one step of antibody hydrolysis within the cell, producing, for example, an amino acid-linker-drug conjugate. This type of linker may be a thioether group or maleimidocaproyl group, and may maintain stability in blood.

[0526] In the present invention, when the disulfide bond chain of the antibody or antigen-binding fragment is reduced, the linker-drug can be bound by introducing a random linkage or an antibody-terminal binding peptide having the sequence GGGGGGGCVIM through a cysteine ​​exposed therein.

[0527] In the present invention, the drug is a pharmacologically effective agent that can be bound to an antibody, and may be, specifically, a chemotherapeutic agent, a toxin, microRNA (miRNA), siRNA, shRNA, or a radioactive isotope. The chemotherapeutic agent may be, for example, a cytotoxic agent or an immunosuppressant. Specifically, it may include a microtubulin inhibitor, a mitotic inhibitor, a topoisomerase inhibitor, or a chemotherapeutic agent that can function as a DNA intercalator. In addition, it may include a drug for treating cancer, inflammation, or immune diseases. For example, it may include an immunomodulatory compound, an anticancer agent, an antiviral agent, an antibacterial agent, an antifungal agent, an anthelmintic agent, or a combination thereof.

[0528] In the present invention, the anticancer agent is, for example, maytansinoid, auristatin (including MMAE, MMAF), aminopterin, actinomycin, bleomycin, talisomycin, camptothecin, N8-acetyl spermidine, 1-(2 chloroethyl)-1,2-dimethyl sulfonyl hydrazide, esperamycin, etoposide, 6-mercaptopurine, dolastatin, trichothecene, calicheamicin, taxol, taxane, paclitaxel, docetaxel, methotrexate, vincristine, vinblastine, doxorubicin, melphalan, mitomycin A, mitomycin C, chlorambucil, duocarmycin, L-asparaginase, mercaptopurine, Thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosourea, cisplatin, carboplatin, mitomycin, dacarbazine, procarbazine, topotecan, nitrogen mustard, cytoxan, etoposide, 5-fluorouracil, bischloroethylnitrosourea, irinotecan, camptothecin, bleomycin, idarubicin, daunorubicin, Dactinomycin, plicamycin, mitoxantrone, asparaginase, vinorelbine, chlorambucil, melphalan, carmustine,Lomustine, busulfan, treosulfan, decarbazine, etoposide, teniposide, topotecan, 9-aminocamptothecin, crisnatol, mitomycin C, trimetrexate, mycophenolic acid, tiazofurin, ribavirin, EICAR (5-ethynyl-1-beta-Dribofuranosylimidazole-4-carboxamide), hydroxyurea, deferoxamine, fluxuridine, doxifluridine, raltitrexed, cytarabine (ara C), cytosine arabinoside, fludarabine, tamoxifen, raloxifene, megestrol, goserelin, leuprolide acetate, flutamide, bicalutamide, EB1089, CB1093, KH1060, verteporfin, phthalocyanine, photosensitizer Pe4, demethoxy-hypocrellin A, interferon-α, interferon-γ, tumor necrosis factor necrosis factor), gemcitabine, velcade, revamid, thalamid, lovastatin,1-methyl-4-phenylpyridinium ion, staurosporine, actinomycin D, dactinomycin, bleomycin A2, bleomycin B2, peplomycin, epirubicin, pirarubicin, zorubicin, mitoxantrone, verapamil, and thapsigargin, nucleases, and toxins of bacterial, animal, or plant origin, but are not limited thereto.

[0529] In the present invention, the anti-inflammatory agent may include, but is not limited to, a steroid preparation that reduces inflammation or swelling by binding to a glucocorticoid receptor, a non-steroidal anti-inflammatory drug (NSAID) that relieves pain by responding to cyclic oxygenase (COX) that synthesizes prostaglandins that cause inflammation, or an immunospecific anti-inflammatory drug (ImSAID) that changes the activation and transport of inflammatory cells.

[0530] In the present invention, the immune disease treatment agent may include, but is not limited to, azathioprine, chlorambucil, cyclophosphamide, cyclosporine, mycophenolate, azathioprine, or methotrexate.

[0531] In the present invention, in some cases, the drug may include one or more nucleophilic groups selected from the group consisting of, but not limited to, amine, thiol, hydroxyl, hydrazide, oxime, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide groups that can react to form a covalent bond with an electrophilic group on a linker and a linker reagent.

[0532]

[0533] chimeric antigen receptor (CAR)

[0534] According to another embodiment of the present invention, there is provided a chimeric antigen receptor (CAR) comprising an antigen binding fragment provided by the present invention.

[0535] As used herein, the term "chimeric antigen receptor" or "CAR" refers to an engineered receptor comprising an extracellular antigen binding domain and an intracellular signaling domain. Although the most common type of CAR comprises a single-chain variable fragment (scFv) derived from a monoclonal antibody fused to the transmembrane and intracellular domains of a T cell coreceptor, such as the CD3ζ chain, the invention described herein is not limited to these domains.

[0536] As used herein, “chimeric antigen receptor” or “CAR” means any receptor engineered to express any intracellular signaling molecule and an extracellular antigen binding domain fused or linked thereto.

[0537] In the present invention, the binding domain may include an antigen binding fragment provided in the present invention, preferably scFv.

[0538] The chimeric antigen receptor of the present invention may further comprise at least one of a hinge region (or spacer) and a signal transduction domain.

[0539] In the present invention, the hinge region is a portion that connects the antigen binding domain and the transmembrane domain, also called a 'spacer', and has the purpose of extending the antigen binding domain from the T cell membrane or NK cell membrane.

[0540] In the present invention, the hinge region may be obtained from any suitable sequence from any genus, including, for example, human or a part thereof, or may include a hinge region of a human protein, including all or part of CD8, D28, 4-1BB, OX40, CD3 zeta (ζ) chain, T cell receptor α or β chain, CD28, CD3ε, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, ICOS, CD154, functional derivatives thereof, or combinations thereof, but is not limited thereto.

[0541] Additionally, in the present invention, the hinge region may include, but is not limited to, one selected from immunoglobulins (e.g., IgG1, IgG2, IgG3, IgG4, and IgD).

[0542] In the present invention, the signal transduction domain refers to a portion of a chimeric antigen receptor that is found or engineered to be found within a T cell. In the present invention, the signal transduction domain may or may not include a transmembrane domain that serves to anchor the chimeric antigen receptor to the plasma membrane of the T cell.

[0543] In the present invention, the transmembrane domain and the signal transduction domain may be derived from the same protein (e.g., CD3ζ), or the transmembrane domain and the signal transduction domain may be derived from different proteins (e.g., the transmembrane domain of CD28 and the intracellular signal transduction domain of the CD3ζ molecule, or vice versa).

[0544] In the present invention, the transmembrane domain comprises a hydrophobic polypeptide that spans the cell membrane. In particular, the transmembrane domain can span from one side of the cell membrane (extracellular) to the other side of the cell membrane (intracellular or cytoplasmic).

[0545] In the present invention, the transmembrane domain may be in the form of an alpha helix, a beta barrel, or a combination thereof. Furthermore, in the present invention, the transmembrane domain may comprise a multi-component protein having multiple transmembrane segments, each of which is in the form of an alpha helix, a beta sheet, or a combination thereof.

[0546] In the present invention, the transmembrane domain may include, but is not limited to, for example, T cell receptor α or β chain, all or part of CD3 zeta (ζ) chain, CD28, CD3ε, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, ICOS, CD154, functional derivatives thereof, or combinations thereof.

[0547] In the present invention, the transmembrane domain may be, for example, an artificially designed polypeptide comprising primarily hydrophobic residues such as leucine and valine. In one embodiment of the present invention, a triad of phenylalanine, tryptophan, and valine may be found at each terminus of the synthetic transmembrane domain.

[0548] In the present invention, at least one costimulatory domain may be further included between the transmembrane domain and the signal transduction domain. In the present invention, the costimulatory domain is a site where a costimulatory signal is transmitted, and may play a role in transmitting a signal so that CAR-T cells or CAR-NK cells that recognize a specific antigen bound to the antigen binding domain cause an immune response, help self-proliferation, and increase the time they remain in the body.

[0549] In the present invention, the costimulatory domain may include, but is not limited to, a functional signaling domain derived from a polypeptide comprising a ligand that binds to 4-1BB (CD137); OX40; CD27; CD28; CD30; CD40; PD-1; CD2; CD7; CD258; natural killer group 2 member C (NKG2C); natural killer group 2 member (NKG2D); B7-H3; CD83; ICAM-1; LFA-1 (CD11a / CD18) or ICOS; an active fragment thereof; a functional inducer thereof; or a combination thereof.

[0550] In the present invention, the signal transduction domain may comprise a polypeptide that provides activation of an immune cell to stimulate or activate at least some aspect of an immune cell signal transduction pathway.

[0551] In the present invention, the signal transduction domain may include, but is not limited to, a functional signal transduction domain derived from a polypeptide comprising all or part of CD3 zeta (ζ), common FcR gamma (FcER1G), FcgammaRIIIa, FcRbeta (Fc epsilon lip), CD3 gamma, CD3 delta, CD3 epsilon, CD79a, CD79b, DNAX-activating protein 10 (DAP10), DNAX-activating protein 12 (DAP12), an active fragment thereof, a functional derivative thereof, or a combination thereof, and such signal transduction domains are known in the art.

[0552] Additionally, the chimeric antigen receptor of the present invention may further comprise a signal peptide.

[0553] In the present invention, the "signal peptide" comprises a peptide sequence, which is a kind of peptide of any secreted or transmembrane protein, and can direct the transport of the chimeric antigen receptor of the present invention to the cell membrane and cell surface, and provide precise localization of the chimeric antigen receptor of the present invention. In particular, in the present invention, the signal peptide supports the chimeric antigen receptor of the present invention to the cell membrane, in which the extracellular portion of the chimeric antigen receptor is displayed on the cell surface, the transmembrane portion spans the plasma membrane, and the active domain is in the cytoplasmic portion, or inside the cell.

[0554] In the present invention, any peptide that performs the above function may be used as the signal peptide without limitation. For example, it may be an n-terminal CD8α signal peptide, but is not limited thereto.

[0555] According to another embodiment of the present invention, the present invention relates to a nucleic acid molecule encoding a chimeric antigen receptor according to the present invention.

[0556] In the present invention, the nucleic acid molecule encoding the chimeric antigen receptor is readily prepared from the amino acid sequence of the specified chimeric antigen receptor by any conventional method. The base sequence encoding the amino acid sequence can be obtained for each domain from the aforementioned NCBI RefSeq ID or GenBenk accession number, and the nucleic acid of the present disclosure can be produced using standard molecular biological and / or chemical procedures. For example, a polynucleotide based on the base sequence can be synthesized, and the nucleic acid molecule of the present disclosure can be produced by combining DNA fragments obtained from a cDNA library using polymerase chain reaction (PCR).

[0557] The nucleic acid molecule of the present invention may be, but is not limited to, a gene or part of an expression or cloning cassette.

[0558]

[0559] According to another embodiment of the present invention, there is provided an expression cassette comprising a nucleic acid molecule provided by the present invention.

[0560] The expression cassette of the present invention may include expression control sequences such as a promoter, enhancer, polyadenylation signal, transcription terminator or internal ribosome entry site (IRES) capable of regulating the expression of the nucleic acid molecule of the present invention.

[0561] In the present invention, the promoter may include, but is not limited to, for example, the SFFV promoter, the elongation factor 1α (EF 1a) promoter, or the CAG (chicken beta-actin promoter with CMV enhancer) promoter.

[0562] The expression cassette of the present invention further comprises a gene sequence encoding a surrogate marker useful for monitoring its expression, for example a cell surface marker (e.g., a truncated cell surface marker), which can be used to confirm transduction or manipulation of cells expressing the receptor.

[0563] Exemplary marker genes in the present invention may include, but are not limited to, base sequences encoding truncated epidermal growth factor receptor (EGFRt), prostate-specific membrane antigen (PSMA), CD34, nerve growth factor receptor (NGFR), or variant forms thereof.

[0564] In addition, in the present invention, the marker gene may include, but is not limited to, a base sequence encoding a fluorescent protein, for example, green fluorescent protein (GFP), enhanced green fluorescent protein (EGFP), for example, super-fold GFP (sfGFP), red fluorescent protein (RFP), for example, tdTomato, mCherry, mStrawberry, AsRed2, DsRed or DsRed2, cyan fluorescent protein (CFP), blue green fluorescent protein (BFP), enhanced blue fluorescent protein (EBFP) and yellow fluorescent protein (YFP), and species variants, monomer variants and codon-optimized and / or enhanced variants of the fluorescent protein.

[0565] In addition, in the present invention, the marker gene is or includes an enzyme, for example, luciferase, lacZ gene derived from Escherichia coli, alkaline phosphatase, secreted embryonic alkaline phosphatase (SEAP), chloramphenicol acetyl transferase (CAT). Exemplary luminescent reporter genes may include, but are not limited to, a base sequence encoding luciferase (luc), β-galactosidase, chloramphenicol acetyltransferase (CAT), β-glucuronidase (GUS), or a variant thereof.

[0566] In addition, in the present invention, the marker gene is a gene encoding a polypeptide that confers resistance to an exogenous agent or drug, for example, a puromycin resistance gene, a hygromycin resistance gene, a blasticidin resistance gene, a neomycin resistance gene, a geneticin resistance gene, or a zeocin resistance gene, or a modified form thereof, or may include the same, but is not limited thereto.

[0567] Additionally, in the present invention, one end of the marker gene and the polynucleotide of the present invention may be linked by a polynucleotide encoding a cleavable peptide.

[0568] In the present invention, the cleavable peptide corresponds to a self-cleavage site and may be, but is not limited to, P2A, F2A, E2A, T2A peptides or a combination thereof.

[0569] According to another embodiment of the present invention, there is provided a recombinant expression vector comprising an expression cassette provided by the present invention.

[0570] In the present invention, the description of the recombinant expression vector overlaps with that described above for the antibody or antigen-binding fragment, and thus the description thereof is omitted below.

[0571] According to another embodiment of the present invention, the present invention relates to an immune effector cell transduced with a recombinant expression vector provided by the present invention.

[0572] In the present invention, the immune effector cells may be allogeneic or autologous immune cells. Furthermore, the immune effector cells in the present invention may be derived from, but are not limited to, T cells, dendritic cells, killer dendritic cells, mast cells, NK cells, NKT cells, macrophages, monocytes, B cells, and stem cells.

[0573] The term "natural killer cells (NK cells)" as used herein is defined as large granular lymphocytes (LGL), which constitute three types of cells that differentiate from common lymphoid progenitor-producing B and T lymphocytes. NK cells are known to differentiate and mature in the bone marrow, lymph nodes, spleen, tonsils, and thymus and enter the circulation. In the present invention, the NK cells may include any type of NK cell without limitation, and may include, but are not limited to, cultured NK cells, such as primary NK cells, NK cells from a cultured NK cell line, or NK cells obtained from a mammal. When the NK cells are obtained from a mammal, the NK cells may be obtained from a number of sources, including but not limited to blood, bone marrow, lymph nodes, thymus, or other tissues or fluids. The NK cells may be concentrated or purified. The NK cells may preferably be human NK cells (e.g., isolated from a human). NK cell lines are available from, for example, the American Type Culture Collection (ATCC), and include, for example, NK-92 cells (ATCC CRL-2407), NK92MI cells (ATCC CRL-2408), or derivatives thereof.

[0574] According to another embodiment of the present invention, the present invention relates to a polypeptide expressed from an immune effector cell provided by the present invention.

[0575]

[0576] therapeutic uses

[0577] According to another embodiment of the present invention, the present invention relates to an antibody or antigen-binding fragment provided by the present invention; or a pharmaceutical composition comprising the antibody or antigen-binding fragment for preventing, improving, or treating a MYCT1-related disease.

[0578] The antibody or antigen-binding fragment of the present invention exhibits excellent affinity and binding affinity for MYCT1. Specifically, it binds specifically to tumor endothelial cells expressing MYCT1 and reprograms the tumor microenvironment (TME), thereby being useful for the treatment of diseases associated with MYCT1 overexpression, such as cancer, and particularly for the treatment of solid tumors.

[0579] The above MYCT1-related disease may be a disease related to the expression or overexpression of MYCT1, and may be, for example, cancer. Accordingly, the MYCT1-related disease is preferably cancer or a tumor, but is not limited thereto.

[0580] As used herein, the term "cancer" refers to or represents a physiological condition typically characterized by uncontrolled cell growth in mammals.

[0581] The cancer or carcinoma that can be treated with the composition of the present invention is not particularly limited and includes both solid cancers and blood cancers. Examples of such cancers may be selected from the group consisting of skin cancer such as melanoma, liver cancer, hepatocellular carcinoma, hepatocellular carcinoma, gastric cancer, breast cancer, lung cancer, ovarian cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colon cancer, colon cancer, pancreatic cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, skin cancer, thyroid cancer, parathyroid cancer, kidney cancer, esophageal cancer, biliary tract cancer, testicular cancer, rectal cancer, head and neck cancer, cervical spine cancer, ureteral cancer, osteosarcoma, neuroblastoma, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, and glioma, but are not limited thereto.

[0582] More preferably, the cancer is characterized by expression of the MYCT1 protein, and may be, but is not limited to, breast cancer, pancreatic cancer, prostate cancer, lung cancer, thyroid cancer, stomach cancer, ovarian cancer, colon cancer, liver cancer, gallbladder cancer, kidney cancer, cervical cancer, or bladder cancer. The cancer may be primary cancer or metastatic cancer.

[0583] In the present invention, the MYCT1-related disease may be an angiogenesis-related disease. The angiogenesis-related disease may be at least one selected from the group consisting of rheumatoid arthritis, osteoarthritis, septic arthritis, psoriasis, corneal ulcer, age-related macular degeneration, diabetic retinopathy, proliferative vitreoretinopathy, immature retinopathy, ophthalmic inflammation, keratoconus, Sjogren's syndrome, myopic ophthalmic tumor, corneal graft rejection, abnormal wound union, bone disease, proteinuria, abdominal aortic aneurysm disease, degenerative cartilage loss due to traumatic joint injury, demyelinating disease of the nervous system, liver cirrhosis, glomerular disease, premature rupture of the gestational membrane, inflammatory bowel disease, periodontal disease, arteriosclerosis, restenosis, inflammatory disease of the central nervous system, Alzheimer's disease, and skin aging, but is not limited thereto.

[0584] The term “angiogenesis” is used herein to encompass all aspects of blood vessel maintenance and development. Therefore, angiogenesis encompasses not only the maintenance and regulation of existing vasculature and small blood vessels, but also the formation of new capillaries, leading to neovascularization. Angiogenesis is a complex process involving a series of sequential steps, including endothelial cell-mediated degradation of the vascular basement membrane and interstitial matrix, endothelial cell migration, endothelial cell proliferation, and endothelial cell-mediated formation of capillary loops. Angiogenesis encompasses the growth and / or development of new blood vessels (also referred to as neovascularization), dilatation of small blood vessels, excessive or prolonged vessel growth, and maintenance of existing vasculature. MYCT1 is known to be involved in the regulation of angiogenesis (see non-patent literature).

[0585] The term "angiogenesis-related disease" is used herein to refer to a specific pathological process in humans in which angiogenesis is abnormally prolonged. This further encompasses angiogenic diseases and conditions, such as diseases and conditions related to, caused by, or associated with angiogenesis. Non-limiting examples of such diseases include various forms of cancer and metastases characterized by angiogenesis / neovascularization. The antibodies and antigen-binding fragments thereof disclosed herein can be used to treat angiogenesis-related diseases by binding to MYCT1 and inhibiting angiogenesis.

[0586] In the present invention, the term "prevention" may include, without limitation, any action that can block, suppress, or delay symptoms caused by a MYCT1-related disease by using the composition of the present invention.

[0587] In the present invention, the terms “improvement” and “treatment” may include, without limitation, any act that can improve or benefit symptoms caused by a MYCT1-related disease by using the composition of the present invention.

[0588] The pharmaceutical composition of the present invention may be formulated and used in the form of oral formulations such as powders, granules, capsules, tablets, aqueous suspensions, etc., external preparations, suppositories, and sterile injection solutions, but is not limited thereto, according to conventional methods. Preferably, the pharmaceutical composition may be formulated for intravenous administration or inhalation; or for use as an injection, but is not limited thereto.

[0589] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may include a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a pigment, a fragrance, etc. for oral administration, and may include a mixture of a buffer, a preservative, an analgesic, a solubilizer, an isotonic agent, a stabilizer, and a stabilizer for injections, and may include a base, an excipient, a lubricant, a preservative, etc. for topical administration. The dosage form of the pharmaceutical composition of the present invention may be prepared in various ways by mixing it with the pharmaceutically acceptable carrier described above. For example, the composition may be prepared in the form of a tablet, troche, capsule, elixir, suspension, syrup, wafer, etc. for oral administration, and the composition may be prepared in the form of a unit dose ampoule or a multiple dose form for injections. In addition, the composition may be formulated as a solution, suspension, tablet, capsule, sustained-release preparation, etc.

[0590] Examples of carriers, excipients, and diluents suitable for the formulation of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, or mineral oil. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, and the like may be additionally included.

[0591] The routes of administration of the pharmaceutical composition of the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal. It can be administered orally or parenterally, and is preferably administered orally, but is not limited thereto.

[0592] The parenteral route of the present invention includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0593] The pharmaceutical composition of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination, and severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, form of medicine, administration route, and period, but may be appropriately selected by a person skilled in the art, and may be administered at 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. The administration may be administered once a day or divided into several times. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, a dragee, a capsule, a liquid, a gel, a syrup, a slurry, or a suspension.

[0594] According to another specific embodiment of the present invention, there is provided a method for preventing, improving or treating a MYCT1-related disease, comprising administering to a subject a therapeutically effective amount of an antibody or antigen-binding fragment provided by the present invention; or a bispecific or multispecific antibody comprising the antibody or antigen-binding fragment.

[0595] In the present invention, the subject is a subject that has developed or is suspected of developing a MYCT1-related disease (particularly a disease caused by overexpression of MYCT1). The subject suspected of developing the disease means all animals including humans, monkeys, cows, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits or guinea pigs that have developed or can develop the disease, but a subject that can be treated with the effective substance provided in the present invention is included without limitation.

[0596] The term "pharmaceutically effective amount" as used herein is an amount sufficient to stop or alleviate the physiological effects of a subject or patient caused by a MYCT1-related disease. An appropriate effective amount may be determined by a treating physician within the scope of sound medical judgment, and may be administered once or in several divided doses. However, for the purpose of the present invention, it is preferable to apply a specific therapeutically effective amount for a specific patient differently depending on various factors such as the type and degree of response to be achieved, whether other agents are used in some cases, the composition containing the specific effective ingredient, the patient's age, weight, general health condition, sex, and diet, the time of administration, the route of administration, the number of administrations, the secretion rate of the composition containing the effective ingredient, the treatment period, drugs used together or concurrently with the specific composition, and similar factors well known in the medical field.

[0597] The total effective amount of the composition of the present invention can be administered to a patient as a single dose, or can be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The composition of the present invention may vary the content of the active ingredient depending on the severity of the disease. Specifically, a preferred total dosage of the composition of the present invention may be about 0.0001 mg to 500 mg per kg of patient body weight per day. However, since the dosage of the composition is determined by taking into consideration various factors such as the route of administration of the pharmaceutical composition and the number of treatments, as well as the patient's age, weight, health status, sex, severity of the disease, diet, and excretion rate, a person having ordinary skill in the art will be able to determine an appropriate effective dosage according to a specific use of the composition of the present invention. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, route of administration, or method of administration as long as it exhibits the effects of the present invention.

[0598]

[0599] Diagnostic purposes

[0600] According to another embodiment of the present invention, the present invention relates to an antibody or antigen-binding fragment thereof; or a composition for diagnosing a MYCT1-related disease comprising the antibody or antigen-binding fragment thereof.

[0601] In the present invention, by detecting MYCT1 by the antibody or antigen-binding fragment, it can be used as a diagnostic marker for evaluating the progression status and prognosis before and after treatment of angiogenesis-related diseases, such as cancer or diseases characterized by angiogenesis.

[0602] The antibodies or antigen-binding fragments of the present invention can be used in any known analytical methods, such as competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays.

[0603] The antibody or antigen-binding fragment of the present invention may be labeled with a label that: (i) provides a detectable signal; (ii) reacts with a first or second label, e.g., a second label to modify a detectable signal provided by FRET (Fluorescence Resonance Energy Transfer); (iii) affects mobility, e.g., electrophoretic mobility, by charge, hydrophobicity, shape, or other physical parameters, or (iv) provides a capture moiety, such as an affinity, antibody / antigen, or ionic complex. Suitable labels include fluorescent labels, luminescent labels, chromophore labels, radioactive labels, isotope labels, isobaric labels, enzymatic labels, particulate labels, in particular metal particle labels, magnetic particle labels, polymer particle labels, small organic molecules such as biotin, receptor ligands or binding molecules such as cell adhesion proteins or lectins, label-sequences comprising nucleic acid and / or amino acid residues that are detectable by use of a binding agent, etc.

[0604] In the present invention, the specific type of the label is not particularly limited, but examples thereof include enzymes that utilize hydrogen peroxide to oxidize a dye precursor, such as HRP, lactoperoxidase or microperoxidase, biotin / avidin, spin labels, bacteriophage labels, radioisotopes 32P, 14C, 125I, 3H and 131I linked to stable free radicals, etc., fluorophores, such as rare earth chelates or fluorescein and derivatives thereof, rhodamine and derivatives thereof, dansyl, umbelliferone, luciferases, such as firefly luciferase and bacterial luciferase (US Patent No. 4,737,456), luciferin, 2,3-dihydrophthalazinedione, horseradish peroxidase (HRP), alkaline phosphatase, β-galactosidase, Glucoamylase, lysozyme, saccharide oxidases such as glucose oxidase, galactose oxidase and glucose-6-phosphate dehydrogenase, heterocyclic oxidases such as uricase and xanthine oxidase, but are not limited thereto.

[0605] According to another embodiment of the present invention, there is provided a diagnostic kit for a MYCT1-related disease comprising the diagnostic composition of the present invention.

[0606] In the present invention, the "kit" refers to a tool that can evaluate the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. It includes not only direct labeling of a detectable substance related to a probe or antibody by reaction with a substrate, but also indirect labeling in which a chromogenic label is conjugated by reactivity with another directly labeled reagent. It may include a chromogenic substrate solution, a washing solution, and other solutions that react with the label to develop a color, and may be manufactured including the reagent components used. In the present invention, the kit may be a kit including the essential elements required for performing RT-PCR, and may include a test tube, a reaction buffer, deoxynucleotides (dNTPs), Taq polymerase, reverse transcriptase, DNase, RNase inhibitor, sterile water, etc. in addition to each primer pair specific for a marker gene. In addition, the kit may be a kit for detecting a gene for cancer diagnosis that includes the essential elements required for performing a DNA chip. The DNA chip kit includes a substrate to which a cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may include a cDNA corresponding to a quantitative control gene or a fragment thereof. The kit of the present invention is not limited thereto, as long as it is known in the art.

[0607] In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

[0608] The kit of the present invention may further comprise one or more other component compositions, solutions or devices suitable for the analysis method. For example, the kit in the present invention may further comprise essential elements necessary for performing a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit comprises a pair of primers specific for a gene encoding a marker protein. The primers are nucleotides having a sequence specific for the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably about 10 bp to 30 bp. It may also comprise a primer specific for the nucleic acid sequence of a control gene. In addition, the reverse transcription polymerase reaction kit may comprise a test tube or other appropriate container, a reaction buffer (with various pH and magnesium concentrations), deoxynucleotides (dNTPs), an enzyme such as Taq polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC-water, sterile water, etc.

[0609] In addition, the diagnostic kit of the present invention may include essential elements necessary for performing a DNA chip. The DNA chip kit may include a substrate to which a cDNA or oligonucleotide corresponding to a gene or a fragment thereof is attached, and reagents, agents, enzymes, etc. for producing a fluorescently labeled probe. In addition, the substrate may include a cDNA or oligonucleotide corresponding to a control gene or a fragment thereof.

[0610] In addition, the diagnostic kit of the present invention may include essential elements necessary for performing ELISA. The ELISA kit includes an antibody specific for the protein. The antibody is an antibody having high specificity and affinity for the marker protein and little cross-reactivity to other proteins, and may be a monoclonal antibody, a polyclonal antibody, or a recombinant antibody. In addition, the ELISA kit may include an antibody specific for a control protein. In addition, the ELISA kit may include reagents capable of detecting bound antibodies, such as labeled secondary antibodies, chromophores, enzymes (e.g., conjugated to antibodies), and their substrates or other substances capable of binding to antibodies.

[0611] In the present invention, a fixative for the antigen-antibody binding reaction may be a nitrocellulose membrane, a PVDF membrane, a well plate synthesized from polyvinyl resin or polystyrene resin, a glass slide glass, etc., but is not limited thereto.

[0612] In the present invention, the washing solution preferably contains a phosphate buffer solution, NaCl, and Tween 20, and a buffer solution (PBST) composed of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20 is more preferred. After the antigen-antibody binding reaction, the washing solution reacts the antigen-antibody complex with a secondary antibody, and then an appropriate amount is added to the fixative and washed 3 to 6 times. The reaction stopping solution can preferably be a sulfuric acid solution (H2SO4).

[0613] According to another specific embodiment of the present invention, there is provided a method for providing information for diagnosing a MYCT1-related disease, comprising the step of contacting a biological sample isolated from a target subject with an antibody or antigen-binding fragment provided in the present invention, or a bispecific or multispecific antibody comprising the antibody or antigen-binding fragment.

[0614] In the present invention, the subject is a subject that has developed or is suspected of developing a MYCT1-related disease (particularly a disease caused by overexpression of MYCT1). The subject suspected of developing the disease means all animals including humans, monkeys, cows, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits or guinea pigs that have developed or can develop the disease. However, any subject that can be diagnosed with a disease using the effective substance provided in the present invention is included without limitation.

[0615] In the present invention, the biological sample is whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, breath, urine, semen, saliva, peritoneal washings, pelvic fluids, cystic fluid, meningeal fluid, amniotic fluid, glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint It may include, but is not limited to, joint aspirate, organ secretions, cells, cell extracts, or cerebrospinal fluid.

[0616] The information providing method of the present invention may include a step of measuring the expression level of MYCT1 protein in the biological sample using the antibody or antigen-binding fragment provided in the present invention.

[0617] In the present invention, measurement of the expression level of the protein can be performed by Western blotting, enzyme linked immunosorbent assay (ELISA), competitive binding assay, or a similar method.

[0618] The information providing method of the present invention may include a step of predicting that a MYCT1-related disease has developed or is likely to develop if the expression level of the MYCT1 protein measured for the biological sample is increased compared to a control group. Here, the control group may be, but is not limited to, the expression level of MYCT1 in a healthy normal control group.

[0619]

[0620] The terms used in the examples are for illustrative purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "comprise" or "have" are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0621] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0622] When describing components of an embodiment, terms such as first, second, A, B, (a), (b), etc. may be used. These terms are only intended to distinguish the components from other components, and the nature, order, or sequence of the components are not limited by the terms. When a component is described as being "connected," "coupled," or "connected" to another component, it should be understood that the component may be directly connected or connected to the other component, but that another component may also be "connected," "coupled," or "connected" between each component.

[0623]

[0624] combination therapy

[0625] According to another specific embodiment of the present invention, there is provided a combination therapy comprising a pharmaceutical composition comprising an antibody or antigen-binding fragment; an immunoconjugate; a chimeric antigen receptor; or an immune effector cell expressing a chimeric antigen receptor as an active ingredient, and at least one additional therapeutic agent or therapeutic procedure.

[0626] The at least one additional therapeutic agent or therapeutic procedure may be selected from, but is not limited to, one or more of chemotherapy, targeted anticancer therapy, oncolytic drug, cytotoxic agent, immune-based therapy, cytokine, surgical procedure, radiation procedure, vaccine, or cell therapy.

[0627] At least one of the above therapeutic agents may be an immunotherapy agent.

[0628] The at least one therapeutic agent may be a PD-1 inhibitor, for example, a PD-1 antibody. Specifically, the PD-1 inhibitor may be selected from, but is not limited to, PDR001, Nivolumab, Pembrolizumab, Pidilizumab, MEDI0680, REGN2810, TSR-042, PF-06801591, or AMP-224.

[0629] At least one of the therapeutic agents may be a PD-L1 inhibitor, such as a PD-L1 antibody. Specifically, the agent may be selected from, but is not limited to, FAZ053, atezolizumab, avelumab, durvalumab, or BMS-936559.

[0630] In the present invention, the term "combination" should be understood to refer to simultaneous, separate, or sequential administration. If the administration is sequential or separate, the interval between administrations of the secondary components should be such that the beneficial effects of the combination are not lost.

[0631]

[0632] Use to enhance sensitivity to immunotherapy

[0633] According to another embodiment of the present invention, the present invention relates to the use of an antibody or antigen-binding fragment provided by the present invention for enhancing the sensitivity of an immunotherapy agent to various cancer types.

[0634] The term "susceptibility" as used herein refers to whether a particular drug (e.g., an immunotherapy) is effective against an individual patient's cancer.

[0635] The term "resistance" as used herein to refer to sensitivity means not reacting sensitively to the drug and thus not taking effect.

[0636] For example, the specific drug mentioned above is primarily an immunotherapy, and these anticancer drugs may or may not be effective depending on the type of cancer. Furthermore, even for cancer types recognized as effective, it is known that individual patients may or may not be effective. Whether an anticancer drug is effective against an individual patient's cancer is referred to as anticancer drug sensitivity. Therefore, if the present invention can predict patients who are expected to respond (responders) and those who are not expected to respond (non-responders) prior to treatment initiation, it is possible to achieve highly effective and safe chemotherapy.

[0637] The present invention provides a method for enhancing the sensitivity of a subject to an immunotherapy agent by administering to the subject a composition comprising a therapeutically effective amount of an antibody or antigen-binding fragment of the present invention.

[0638]

[0639] Hereinafter, embodiments are described in detail with reference to the attached drawings. However, the embodiments may be modified in various ways, and the scope of the patent application is not limited or restricted by these embodiments. It should be understood that all modifications, equivalents, or alternatives to the embodiments are included within the scope of the patent application.

[0640] In addition, when describing with reference to the attached drawings, identical components will be assigned the same reference numerals regardless of the drawing numbers, and redundant descriptions thereof will be omitted. When describing embodiments, if a detailed description of a related known technology is judged to unnecessarily obscure the gist of the embodiment, the detailed description will be omitted.

[0641]

[0642] The present invention is susceptible to various modifications and embodiments. Specific embodiments are illustrated in the drawings and described in detail in the following detailed description. However, this is not intended to limit the present invention to specific embodiments, but rather to encompass all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention. In describing the present invention, detailed descriptions of related known technologies will be omitted if they are deemed to obscure the gist of the present invention.

[0643]

[0644] Example 1. Screening of phage display libraries

[0645] 1-1. Preparation of antigen

[0646] The antigen used for screening the phage display library for the production of anti-MYCT1 antibodies was synthesized by WuXi Biologics, and included amino acid sequences 1 to 235 of human MYCT1 (SEQ ID NO: 1), amino acid sequences 1 to 188 of mouse MYCT1 (SEQ ID NO: 2), and the overlapping sequence of the mouse transmembrane domain (SEQ ID NO: 3). Amino acid sequences 49 to 67 of human MYCT1 and 1 to 19 of mouse MYCT1 were used for screening by conjugating biotin. The results of comparison of the amino acid sequences of human and mouse MYCT1 are shown in Fig. 1.

[0647]

[0648] 1-2. Antibody selection through phage library screening for antigen production

[0649] Human synthetic antibody library (Synthetic Fab) and human unprimed antibody library (Na) provided by WuXi Biologics We performed three rounds of panning using two types of libraries with the scFv gene, and the enrichment titer was confirmed in each round through polyclonal phage ELISA. As the number of rounds increased, the phage for the target antigen was enriched, and the titer was confirmed to increase. (Results not shown).

[0650]

[0651] 1-3. Selection of monophages with binding specificity for MYCT1

[0652] To detect phages binding to MYCT1, a monoclonal phage ELISA (Enzyme-Linked Immunosorbent Assay) was performed. Specifically, MYCT1 (positive) and negative antigens were suspended in DPBS at μg / mL and added to an immunoplate at 100 μL / well to coat the antigen. The supernatant was removed, and blocking solution (4% skim milk) was added to block. The supernatant was removed, and 100 μL each of poly-phage (or mono-phage) was added to the antigen-coated immunoplate to induce binding. The supernatant was removed, and 200 μL / well of PBST was added three times to wash. The immunoplate was reacted with 100 μL / well of anti-M13-HRP. The supernatant was removed, and 200 μL / well of PBST was added three times to wash. 100 μL / well of TMB peroxidase substrate (or o-phenylenediamine dihydrochloride) was added to the immunoplate, and the reaction was stopped after 10 minutes by adding 50 μL / well of stop solution (with added sulfuric acid). The absorbance was measured at a wavelength of 450 nm (or 490 nm) using an ELISA device (plate reader). scFv na Eighty-seven antibodies were selected from the ve library and 29 antibodies were selected from the Fab synthetic library.

[0653]

[0654] 1-4. Selection of phages with binding specificity for cells expressing MYCT1

[0655] Four types of cells were used for cell binding analysis: WT MCEC (Mouse cardiac endothelial cell line), mMOE-FLAG MCEC (mouseMYCT1-FLAG tagged overexpressed Mouse cardiac endothelial cell line), WT iMLEC (Mouse lung endothelial cell line), and iMLEC mKO hMOE long FLAG (MYCT1 knockout mouse lung endothelial cell line [CSII-MCS-IRES2-bsr.FLAG-humanMYCT1]). 200,000 cells were placed in a 96-well plate, phage was diluted two-fold with 1% BSA / 1X PBS, reacted at 4°C for 1 hour, anti-c-Myc-RPE antibody was diluted 1:800 with 1% BSA / 1X PBS, reacted in the dark at 4°C for 30 minutes, dissolved in 100 μL PBS, and FACS analysis was performed to select 23 antibodies that bind to human hyperlipidemia cell lines.

[0656] The CDR sequences of the antibodies selected in Examples 1-3 are as shown in Tables 1 to 8 below, and the sequences of the heavy chain variable region and light chain variable region are organized as shown in Tables 9 to 22 below.

[0657]

[0658] [Table 1]

[0659]

[0660]

[0661] [Table 2]

[0662]

[0663]

[0664] [Table 3]

[0665]

[0666]

[0667] [Table 4]

[0668]

[0669]

[0670] [Table 5]

[0671]

[0672] [Table 6]

[0673]

[0674]

[0675] [Table 7]

[0676]

[0677]

[0678] [Table 8]

[0679]

[0680]

[0681] [Table 9]

[0682]

[0683]

[0684] [Table 10]

[0685]

[0686]

[0687] [Table 11]

[0688]

[0689]

[0690] [Table 12]

[0691]

[0692]

[0693] [Table 13]

[0694]

[0695]

[0696] [Table 14]

[0697]

[0698]

[0699] [Table 15]

[0700]

[0701]

[0702] [Table 16]

[0703]

[0704]

[0705] [Table 17]

[0706]

[0707]

[0708] [Table 18]

[0709]

[0710]

[0711] [Table 19]

[0712]

[0713]

[0714] [Table 20]

[0715]

[0716]

[0717] [Table 21]

[0718]

[0719]

[0720] [Table 22]

[0721]

[0722]

[0723] Example 2. Conversion of anti-MYCT1 scFv and Fab into full IgG form and production thereof

[0724] 2-1. Cloning of anti-MYCT1 scFv and Fab into full IgG form

[0725] Polynucleotides having nucleotide sequences encoding the scFv and Fab antibodies selected in Example 1 were synthesized. The prepared polynucleotides were cloned into animal cell expression vectors (heavy chain expression vector: pcDNA3.4-HC, and light chain expression vector: pcDNA3.4-LC). A total of 46 vectors containing polynucleotides encoding the heavy and light chains for each of the 23 antibody clones in Table 7 were prepared. Each of the vectors prepared for pcDNA3.4-HC and pcDNA3.4-HC contained an IgG1-type sequence.

[0726]

[0727] 2-2. Expression and culture of anti-MYCT1 antibodies

[0728] The expression of anti-MYCT1 antibody converted into whole IgG form of scFv and Fab of Example 2-1 was performed using ExpiCHO-S cells developed by Thermo Fisher (Thermo Fisher, A29127) and following the protocol of ExpiCHO Expression System Kit (Thermo Fisher, A29133). Specifically, ExpiCHO-S cells were cultured under conditions of 37°C, 80% humidity, 8% CO2, and 120 rpm, and on the day of transfection, 6X10 ExpiCHO-S cells were cultured. 6ExpiCHO-S expression medium (Thermo Fisher, A29100-01) was added to dilute and prepared at a cell concentration of 10 cells / mL. The heavy chain vector of Example 2-2 was diluted in OptiPRO SFM medium (Thermo Fisher, 12309050) at 0.5 μg per mL of medium, and the light chain vector of Example 2-2 was diluted in OptiPRO SFM medium at 0.5 μg per mL, and the ExpiFectamine CHO Reagent was diluted in OptiPRO SFM medium. The vector and ExpiFectamine CHO mixture were mixed and reacted at room temperature for 1 to 5 minutes, and then cultured for 20 hours at 37°C, 80% humidity, 8% CO2, and 120 rpm. After 20 hours, ExpiCHO Enhancer and ExpiCHO Feed from the ExpiCHO Expression System Kit (Thermo Fisher, A29133) were added and cultured for 10 days. After culture, the cell culture solution was centrifuged for 30 minutes at 4°C and 3,600 rpm, and the supernatant was separated and stored in the refrigerator.

[0729]

[0730] 2-3. Confirmation of anti-MYCT1 antibody expression

[0731] The culture status and expression of the antibody converted into the full IgG form of the anti-MYCT1 scFv and Fab of Example 2-2 were confirmed by measuring the viable cell density and viability, and the titer was measured through Protein A HPLC.

[0732] Specifically, 10 μL of the culture medium was sampled on days 0, 1, 3, 6, and 7, and the viable cell density and viability were measured using the EVE PLUS equipment. To measure the titer via Protein A HPLC, the TSKgel Protein A-5PW column was equilibrated with 6 mL of equilibration buffer (20 mM sodium phosphate, pH 7.5) at a flow rate of 2 mL / min. 50 μL each of 0.03, 0.06, 0.13, 0.25, 0.5, 1, and 2 mg / mL of human IgG were injected into the TSKgel Protein A-5PW column, and the equilibration buffer was flowed at a flow rate of 2 mL / min for 0.5 min to bind, and then the antibody was eluted by flowing the elution buffer (20 mM sodium phosphate, pH 2.5) for 0.6 min. A calibration curve was drawn with the extracted peaks and R 2 The value was confirmed to be 0.9998, and the titer was calculated using the calibration curve. The culture medium stored in Example 2-2 was filtered using a 0.22 μm PES Bottle Top Filter. 50 μL of the filtered sample was applied to a TSKgel Protein A-5PW column using the same method to elute the antibody. The titer of the eluted peak was calculated using the calibration curve, and the results are shown in Figures 2a, 2b, 2c, and 2d and Figures 3a, 3b, 3c, and 3d.

[0733] Figures 2a, 2b, 2c, 2d and Figures 3a, 3b, 3c, 3d show the results of measuring the viable cell density and viability of ExpiCHO-S cells transfected with a vector containing the sequence of an antibody converted into the full IgG form of anti-MYCT1 scFv and Fab.

[0734] As shown in Figs. 2a, 2b, 2c, 2d and Figs. 3a, 3b, 3c, 3d, it was confirmed that the viable cell density and viability of cells (ExpiCHO-S cells) transformed with the vector containing the antibody sequence of Example 2-2 were high. In addition, the titer of the expressed antibody was measured through Protein A HPLC. This means that the antibody converted into the whole IgG form of the anti-MYCT1 scFv and Fab of Example 2-2 was well expressed in the cells.

[0735] Figure 4 shows the results of measuring the Protein A HPLC titer of a vector containing the sequence of an antibody converted into the full IgG form of anti-MYCT1 scFv and Fab.

[0736]

[0737] 2-4: Isolation and purification of anti-MYCT1 antibodies

[0738] To isolate and purify the anti-MYCT1 antibody of Example 2-2, the culture medium stored in Example 2-2 was filtered through a 0.22 μm PES Bottle Top Filter. 5 mL of Amsphere A3 resin was packed into a Hiscale 16 / 20 column, washed with 100 mL of triple-distilled water at a flow rate of 5 mL / min, and the column was equilibrated with 25 mL of equilibration buffer (0.1 M Tris, pH 7.0). The filtered sample was bound to the Amsphere A3 column at a flow rate of 1.5 mL / min, and then 10 mL of equilibration buffer was flowed to stabilize the UV280 nm value to the baseline. Impurities were removed by flowing 15 mL of wash buffer I (0.1 M Tris, 1 M NaCl, pH 6.0) at a flow rate of 5 mL / min, and the UV280 nm value was stabilized to the Base Line by flowing 15 mL of wash buffer II (0.1 M Tris, pH 6.0). The antibody was eluted by flowing 25 mL of elution buffer (0.1 M Glycine, pH 3.5) at a flow rate of 1.5 mL / min, and the fraction volume was set to 4 mL, and 600 μL of saturated Tris solution was added to each fraction tube. The eluted antibody was placed in a dilution tube and exchanged with 3 L of PBS, pH 7.4 buffer in a low temperature room (4°C) for 4 hours, and then replaced with 3 L of new PBS, pH 7.4 buffer and exchanged for 16 hours. After filtration with a 0.22 μm PES syringe filter, the A280 absorbance was measured to calculate the concentration, and the results are shown in Fig. 5.

[0739] Figures 5a, 5b, 5c, 5d, 5e and 5f are chromatograms showing the results of separating and purifying antibodies converted into full IgG forms of anti-MYCT1 scFv and Fab.

[0740] As shown in Figures 5a, 5b, 5c, 5d, 5e, and 5f, antibodies converted to whole IgG forms of MYCT1 Fab and scFv were confirmed to be well isolated and purified from cell culture media. Among 23 candidates, only 22 were isolated and purified.

[0741]

[0742] 2-5. Confirmation of the purity of the isolated and purified anti-MYCT1 antibody

[0743] To confirm the purity of the anti-MYCT1 antibody of Example 2-4, SDS-PAGE and SEC-HPLC were performed.

[0744] Specifically, the anti-MYCT1 antibody of Example 2-4, non-reducing sample and reducing sample were prepared respectively. The non-reducing sample was prepared by adding 6 μL of 4X LDS sample buffer to 2 μg / 18 μL, vortexing at 3,300 rpm for 10 seconds, and centrifuging for 1 minute. The reducing sample was prepared by adding 6 μL of 4X LDS sample buffer and 2.5 μL of 10X sample reducing reagent to 2 μg / 15.5 μL, vortexing at 3,300 rpm for 10 seconds, heating on a heating block at 100 ℃ for 10 minutes, and centrifuging for 1 minute. Each sample was loaded with 5 μL of protein marker and 24 μL of test solution onto a 4-12% Bis-Tris Mini Protein Gel and electrophoresis was performed at 140 V for 60 minutes. After electrophoresis, 20 mL of EZ-Gel Staining Solution was added, stained for 1 hour, destained with triple-distilled water, and the bands were observed. To confirm the purity by SEC-HPLC, a TSKgel G3000SWXL Column was used, and a 10 μg sample was injected, and 50 mM sodium phosphate, 150 mM NaCl, pH 7.0 was flowed at a flow rate of 1 mL / min for 30 minutes, and the UV214 nm value was measured to analyze the purity (SEC-HPLC), and the results are shown in Figs. 6 to 10. Figs. 6a, 6b, 6c, Figs. 7a, 7b, 7c, 7d, and 8 show the results of confirming the purity (SDS-PAGE) of the anti-MYCT1 antibody of Example 2-4. Table 23 shows the results of confirming the purity (SEC-HPLC) of the anti-MYCT1 antibody of Example 2-4. As shown in Table 23 and FIGS. 6a, 6b, 6c, 7a, 7b, 7c, 7d, and 8, it was confirmed that the purity of the anti-MYCT1 antibody of Example 2-4 was high.

[0745] [Table 23]

[0746]

[0747]

[0748] Example 3. Analysis of binding specificity of anti-MYCT1 antibodies to MYCT1

[0749] 3-1. Cloning of anti-MYCT1 scFv and Fab into full IgG form

[0750] The specific binding affinity of the purified anti-MYCT1 antibody from Example 2-4 to the synthetic peptide MYCT1 antigen was analyzed using an ELISA method. Specifically, the antigen protein used was a domain corresponding to the extracellular region of human MYCT1, and biotin was conjugated to the N-terminus and C-terminus, respectively. Streptavidin was used together to effectively expose the antigen epitope. The test was performed as follows using an HRP-conjugated anti-human polyclonal antibody reagent (Sigma, cat# A8667-2ML) as a secondary antibody.

[0751] 50 nM Streptavidin and 50 nM synthetic peptide antigen MYCT1 per well were coated on an ELISA plate, and 3% BSA was incubated to prevent nonspecific binding, and the plate was washed with PBS-T (0.05%). The prepared anti-MYCT1 antibody was added to each well and serially diluted 12 times in 3-fold steps from a concentration of 100 nM, incubated for 1 hour at room temperature, and the plate was washed three times with PBS-T. After color development using HRP-conjugated anti-human polyclonal antibody and TMB (Sigma, cat# T4319-4X100ML), the absorbance was measured at 450 nm and the results were analyzed, and the results are shown in Fig. 11.

[0752] Figures 9a, 9b, and 9c are graphs showing the results of ELISA analysis of the specific binding affinity of purified anti-MYCT1 antibodies to the synthetic peptide antigen MYCT1. The top 12 antibodies with the highest binding affinity were selected from 22 anti-MYCT1 antibodies.

[0753]

[0754] 3-2. Measurement of binding ability of anti-MYCT1 antibodies to cell surface-expressed MYCT1 antigen

[0755] The binding of selected anti-MYCT1 antibodies to MYCT1 expressed on the cell surface was analyzed using a FACS system. Specifically, MLEC-HA cells overexpressing human MYCT1 were prepared and analyzed in comparison with primary HUVEC cells.

[0756] The cells prepared above were added with 300 nM of the purified anti-MYCT1 antibody in Example 2-4, incubated at 4°C for 1 hour, and washed twice with PBS. The anti-human IgG antibody-APC was diluted to 10 nM, incubated at 4°C for 30 minutes, and washed three times with PBS. The fluorescence intensity of the cells was measured using a CytoFLEX S (Beckman) device, and the results are shown in Figs. 10a, 10b, 10c, 10d, 10e, 10f, and 10g.

[0757] Figures 10a, 10b, 10c, 10d, 10e, 10f and 10g are graphs showing the results of analyzing the extent to which the purified anti-MYCT1 antibodies in Example 2-4 bind to MYCT1 expressed on the cell surface using a FACS system.

[0758]

[0759] 3-3. SPR full dynamics analysis of anti-MYCT1 antibodies

[0760] SPR full kinetic analysis was performed to determine the binding affinity of the selected anti-MYCT1 antibodies to the human MYCT1 peptide. The SA sensor chip used in the SPR analysis is based on the biotinylated ligand capture method, and the binding affinity was evaluated by immobilizing the MYCT1 peptide ligand at the level of 28 RU. The binding constant (K) for each antibody a , K d , K D ) was derived and data reliability was evaluated, and the example was used to quantify the specific binding affinity between antigen and antibody and to prove the superiority of the antibody.

[0761] Specifically, a biotinylated MYCT1 peptide was used as the ligand, and 10 antibodies were selected again from the 12 types with high ELISA binding affinity in Example 3-1 and used as analytes. The running buffer used for the analysis was HBS-EP+ pH 7.4 buffer, and the regeneration buffer was 10 mM glycine-HCl pH 2.0 buffer. The injection conditions were a contact time of 180 s, a dissociation time of 240 s, and a flow rate of 30 μL / min, and the analysis was based on a 1:1 binding model. Each binding constant was within the reliable statistical indicators Chi2 value (below 10% of Rmax) and U-value (Not above~25), and binding affinity results were obtained for all antibodies except the P2R31-H3 antibody, which appeared to bind nonspecifically to the SA sensor chip. The results are shown in Table 24 and Figures 11a, 11b, and 11c.

[0762]

[0763] [Table 24]

[0764]

[0765]

[0766] Example 4. Evaluation of the anti-MYCT1 antibody's antiangiogenesis efficacy (Migration Assay)

[0767] Human umbilical vein endothelial cells (HUVEC) (Lonza, Switzerland) were cultured in EGM-2 (endothelial growth medium, Lonza, Switzerland) supplemented with 10% fetal bovine serum (FBS), insulin (5 μg / mL), recombinant human basic fibroblast growth factor (rhFGF; 1 μg / mL), recombinant human epithermal growth factor (rhEGF; 0.1 μg / mL), gentamicin (50 μg / mL), and amphotericin B (50 μg / mL), and passage 4 to 5 cells were used. To harvest the cells, they were washed with PBS, treated with 0.25% trypsin-EDTA (Gibco, USA), and after 2 minutes, M199 (Lonza, Switzerland) containing 10% fetal bovine serum was added to neutralize the enzyme action. After detachment, the cells were collected, centrifuged at 1,700 rpm for 3 minutes, diluted to the desired cell number by adding EGM-2, and suspended. The wound-healing assay was then performed according to the manufacturer's protocol (Ibidi, Martinsried, Germany). Specifically, cells were seeded in each well, and all experimental groups except the control were treated with 50 ng / mL of recombinant human VEGF, and each well was treated with 300 nM of the anti-MYCT1 antibody purified in Example 2-4. After culturing for 24 hours, microscopic images were taken and evaluated using image analysis with the image analysis program Image J (NIH, USA). The group treated with bevacizumab served as a positive control. The results are shown in Figures 12a, 12b and 13a and 13b.

[0768] As shown in Figures 12a, 12b and 13a and 13b, the evaluated anti-MYCT1 human antibodies were confirmed to have an ability to inhibit cell motility equivalent to or greater than that of the control group.

[0769]

[0770] Although the embodiments described above have been described with limited drawings, those skilled in the art will appreciate that various technical modifications and variations can be applied based on the above. For example, appropriate results can still be achieved even if the described techniques are performed in a different order than described, and / or components of the described systems, structures, devices, circuits, etc. are combined or combined in a different manner than described, or are replaced or substituted with other components or equivalents.

[0771] Therefore, other implementations, other embodiments, and equivalents to the claims also fall within the scope of the claims described below.

Claims

1. A heavy chain CDR1 (HCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 43; A heavy chain CDR2 (HCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 44 to 85; A heavy chain CDR3 (HCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 187; A light chain CDR1 (LCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 188 to 264; A light chain CDR2 (LCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 265 to 320; and An anti-MYCT1 human antibody specific for human MYCT1, or an antigen-binding fragment thereof, comprising a light chain CDR3 (LCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 321 to 411.

2. In paragraph 1, The above HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2 and LCDR-3 are, in order, anti-MYCT1 human antibodies or antigen-binding fragments thereof, each comprising a CDR combination of the amino acid sequences set forth below: SEQ ID NOs: 13, 76, 105, 222, 301, and 346; SEQ ID NOs: 40, 53, 160, 262, 278, and 404; SEQ ID NOs: 42, 56, 113, 262, 283, and 407; SEQ ID NOs: 26, 44, 147, 201, 266, and 358; SEQ ID NOs: 25, 77, 110, 251, 293, and 388; SEQ ID NOs: 6, 68, 143, 230, 304, and 325; SEQ ID NOs: 25, 77, 109, 252, 293, and 382; SEQ ID NOs: 42, 56, 112, 262, 283, and 409; SEQ ID NOs: 40, 52, 160, 245, 279, and 401; SEQ ID NOs: 42, 56, 98, 233, 272, and 338; SEQ ID NOs: 42, 56, 95, 237, 310, and 324; SEQ ID NOs: 25, 77, 114, 246, 275, and 403; SEQ ID NOs: 25, 77, 126, 251, 293, and 386; SEQ ID NOs: 30, 48, 130, 194, 320, and 378; SEQ ID NOs: 25, 77, 128, 249, 307, and 328; SEQ ID NOs: 33, 72, 91, 205, 296, and 368; SEQ ID NOs: 42, 56, 156, 222, 301, and 346; SEQ ID NOs: 16, 63, 131, 262, 282, and 397; SEQ ID NOs: 25, 77, 125, 251, 293, and 384; SEQ ID NOs: 42, 56, 118, 264, 277, and 392; SEQ ID NOs: 25, 77, 117, 251, 293, and 387; SEQ ID NOs: 42, 56, 95, 239, 310, and 324; SEQ ID NOs: 20, 74, 138, 200, 297, and 354; SEQ ID NOs: 6, 65, 123, 193, 319, and 373; SEQ ID NOs: 42, 56, 95, 239, 310, and 322; SEQ ID NOs: 42, 59, 90, 254, 279, and 395; SEQ ID NOs: 42, 56, 94, 239, 310, and 325; SEQ ID NOs: 42, 56, 88, 261, 276, and 393; SEQ ID NOs: 8, 66, 116, 189, 315, and 340; SEQ ID NOs: 42, 56, 133, 260, 295, and 394; SEQ ID NOs: 42, 56, 87, 241, 290, and 334; SEQ ID NOs: 24, 79, 137, 262, 283, and 405; SEQ ID NOs: 7, 64, 186, 227, 273, and 342; SEQ ID NOs: 30, 48, 129, 197, 319, and 378; SEQ ID NOs: 25, 77, 99, 251, 293, and 387; SEQ ID NOs: 25, 77, 164, 203, 296, and 369; SEQ ID NOs: 42, 56, 163, 250, 294, and 383; SEQ ID NOs: 34, 54, 146, 229, 303, and 333; SEQ ID NOs: 39, 81, 127, 244, 312, and 335; SEQ ID NOs: 5, 67, 144, 228, 310, and 321; SEQ ID NOs: 26, 44, 150, 235, 309, and 330; SEQ ID NOs: 42, 56, 104, 247, 293, and 385; SEQ ID NOs: 27, 77, 172, 262, 278, and 408; SEQ ID NOs: 42, 56, 101, 195, 319, and 379; SEQ ID NOs: 26, 44, 161, 239, 310, and 324; SEQ ID NOs: 42, 56, 97, 197, 319, and 375; SEQ ID NOs: 43, 80, 149, 243, 317, and 323; SEQ ID NOs: 27, 51, 152, 210, 268, and 357; SEQ ID NOs: 25, 78, 119, 231, 293, and 387; SEQ ID NOs: 15, 47, 107, 215, 301, and 341; SEQ ID NOs: 42, 56, 102, 258, 274, and 402; SEQ ID NOs: 35, 61, 151, 214, 301, and 347; SEQ ID NOs: 42, 56, 95, 239, 310, and 325; SEQ ID NOs: 8, 66, 116, 188, 314, and 340; SEQ ID NOs: 14, 85, 153, 220, 299, and 349; SEQ ID NOs: 39, 81, 127, 244, 313, and 335; SEQ ID NOs: 42, 57, 132, 191, 305, and 389; SEQ ID NOs: 36, 75, 121, 236, 280, and 337; SEQ ID NOs: 25, 77, 168, 221, 301, and 345; SEQ ID NOs: 31, 56, 148, 197, 319, and 378; SEQ ID NOs: 42, 56, 106, 256, 285, and 396; SEQ ID NOs: 26, 44, 167, 255, 284, and 398; SEQ ID NOs: 42, 56, 95, 190, 310, and 324; SEQ ID NOs: 6, 65, 124, 197, 319, and 373; SEQ ID NOs: 17, 62, 120, 196, 319, and 380; SEQ ID NOs: 4, 65, 166, 226, 318, and 336; SEQ ID NOs: 42, 57, 96, 239, 310, and 325; SEQ ID NOs: 25, 77, 154, 217, 302, and 343; SEQ ID NOs: 42, 56, 115, 263, 281, and 411; SEQ ID NOs: 32, 49, 108, 225, 307, and 410; SEQ ID NOs: 42, 56, 95, 242, 308, and 331; SEQ ID NOs: 26, 44, 150, 234, 316, and 326; SEQ ID NOs: 42, 56, 95, 192, 311, and 322; SEQ ID NOs: 26, 44, 155, 235, 309, and 330; SEQ ID NOs: 25, 77, 168, 213, 301, and 345; SEQ ID NOs: 24, 79, 137, 262, 283, and 406; SEQ ID NOs: 42, 56, 100, 248, 307, and 391; SEQ ID NOs: 26, 44, 183, 197, 319, and 378; SEQ ID NOs: 25, 77, 93, 251, 293, and 387; SEQ ID NOs: 42, 55, 103, 259, 283, and 400; SEQ ID NOs: 42, 56, 92, 262, 278, and 399; SEQ ID NOs: 26, 44, 170, 198, 288, and 352; SEQ ID NOs: 25, 77, 165, 251, 291, and 387; SEQ ID NOs: 42, 56, 102, 257, 274, and 402; SEQ ID NOs: 25, 77, 111, 253, 292, and 381; SEQ ID NOs: 38, 58, 86, 219, 301, and 348; SEQ ID NOs: 42, 56, 122, 262, 283, and 405; SEQ ID NOs: 42, 56, 134, 209, 289, and 364; SEQ ID NOs: 32, 50, 162, 216, 300, and 344; SEQ ID NOs: 20, 69, 141, 211, 269, and 356; SEQ ID NOs: 12, 73, 140, 199, 297, and 353; SEQ ID NOs: 28, 70, 145, 218, 300, and 390; SEQ ID NOs: 21, 74, 135, 210, 267, and 370; SEQ ID NOs: 20, 45, 177, 210, 289, and 363; SEQ ID NOs: 28, 46, 184, 204, 287, and 366; SEQ ID NOs: 20, 45, 175, 209, 265, and 350; SEQ ID NOs: 26, 74, 178, 209, 286, and 377; SEQ ID NOs: 21, 74, 187, 209, 268, and 374; SEQ ID NOs: 19, 44, 142, 200, 297, and 355; SEQ ID NOs: 23, 74, 174, 211, 268, and 351; SEQ ID NOs: 10, 74, 89, 209, 289, and 372; SEQ ID NOs: 37, 60, 139, 206, 286, and 339; SEQ ID NOs: 41, 83, 157, 232, 306, and 329; SEQ ID NOs: 18, 74, 178, 209, 268, and 372; SEQ ID NOs: 32, 49, 179, 224, 300, and 344; SEQ ID NOs: 20, 74, 176, 210, 287, and 359; SEQ ID NOs: 26, 74, 178, 209, 287, and 361; SEQ ID NOs: 22, 71, 171, 210, 286, and 360; SEQ ID NOs: 11, 44, 159, 208, 286, and 371; SEQ ID NOs: 20, 44, 185, 212, 287, and 365; SEQ ID NOs: 41, 84, 136, 238, 271, and 332; SEQ ID NOs: 20, 44, 173, 202, 298, and 367; SEQ ID NOs: 20, 74, 182, 209, 286, and 376; SEQ ID NOs: 9, 74, 180, 207, 286, and 362; SEQ ID NOs: 41, 82, 158, 240, 270, and 327; ; or SEQ ID NOs: 29, 44, 181, 209, 286, and 376.

3. In paragraph 1, An anti-MYCT1 human antibody or antigen-binding fragment thereof comprising at least one variable heavy region (VH) selected from the group consisting of SEQ ID NOs: 412 to 527.

4. In paragraph 1, An anti-MYCT1 human antibody or antigen-binding fragment thereof comprising at least one variable light chain region (VL) selected from the group consisting of SEQ ID NOs: 528 to 643.

5. In paragraph 1, Containing the VH sequence of SEQ ID NO: 412 and the VL sequence of SEQ ID NO: 528; Containing the VH sequence of SEQ ID NO: 413 and the VL sequence of SEQ ID NO: 529; Containing the VH sequence of SEQ ID NO: 414 and the VL sequence of SEQ ID NO: 530; Containing the VH sequence of SEQ ID NO: 415 and the VL sequence of SEQ ID NO: 531; Containing the VH sequence of SEQ ID NO: 416 and the VL sequence of SEQ ID NO: 532; Containing the VH sequence of SEQ ID NO: 417 and the VL sequence of SEQ ID NO: 533; Containing the VH sequence of SEQ ID NO: 418 and the VL sequence of SEQ ID NO: 534; Containing the VH sequence of SEQ ID NO: 419 and the VL sequence of SEQ ID NO: 535; Containing the VH sequence of SEQ ID NO: 420 and the VL sequence of SEQ ID NO: 536; Containing the VH sequence of SEQ ID NO: 421 and the VL sequence of SEQ ID NO: 537; Containing the VH sequence of SEQ ID NO: 422 and the VL sequence of SEQ ID NO: 538; Containing the VH sequence of SEQ ID NO: 423 and the VL sequence of SEQ ID NO: 539; Containing the VH sequence of SEQ ID NO: 424 and the VL sequence of SEQ ID NO: 540; Containing the VH sequence of SEQ ID NO: 425 and the VL sequence of SEQ ID NO: 541; Containing the VH sequence of SEQ ID NO: 426 and the VL sequence of SEQ ID NO: 542; Containing the VH sequence of SEQ ID NO: 427 and the VL sequence of SEQ ID NO: 543; Containing the VH sequence of SEQ ID NO: 428 and the VL sequence of SEQ ID NO: 544; Containing the VH sequence of SEQ ID NO: 429 and the VL sequence of SEQ ID NO: 545; Containing the VH sequence of SEQ ID NO: 430 and the VL sequence of SEQ ID NO: 546; Containing the VH sequence of SEQ ID NO: 431 and the VL sequence of SEQ ID NO: 547; Containing the VH sequence of SEQ ID NO: 432 and the VL sequence of SEQ ID NO: 548; comprising the VH sequence of SEQ ID NO: 433 and the VL sequence of SEQ ID NO: 549; or An anti-MYCT1 human antibody or antigen-binding fragment thereof, comprising the VH sequence of SEQ ID NO: 434 and the VL sequence of SEQ ID NO:

550.

6. In paragraph 1, Containing the VH sequence of SEQ ID NO: 435 and the VL sequence of SEQ ID NO: 551; Containing the VH sequence of SEQ ID NO: 436 and the VL sequence of SEQ ID NO: 552; Containing the VH sequence of SEQ ID NO: 437 and the VL sequence of SEQ ID NO: 553; Containing the VH sequence of SEQ ID NO: 438 and the VL sequence of SEQ ID NO: 554; Containing the VH sequence of SEQ ID NO: 439 and the VL sequence of SEQ ID NO: 555; Containing the VH sequence of SEQ ID NO: 440 and the VL sequence of SEQ ID NO: 556; Containing the VH sequence of SEQ ID NO: 441 and the VL sequence of SEQ ID NO: 557; Containing the VH sequence of SEQ ID NO: 442 and the VL sequence of SEQ ID NO: 558; Containing the VH sequence of SEQ ID NO: 443 and the VL sequence of SEQ ID NO: 559; Containing the VH sequence of SEQ ID NO: 444 and the VL sequence of SEQ ID NO: 560; Containing the VH sequence of SEQ ID NO: 445 and the VL sequence of SEQ ID NO: 561; Containing the VH sequence of SEQ ID NO: 446 and the VL sequence of SEQ ID NO: 562; Containing the VH sequence of SEQ ID NO: 447 and the VL sequence of SEQ ID NO: 563; Containing the VH sequence of SEQ ID NO: 448 and the VL sequence of SEQ ID NO: 564; Containing the VH sequence of SEQ ID NO: 449 and the VL sequence of SEQ ID NO: 565; Containing the VH sequence of SEQ ID NO: 450 and the VL sequence of SEQ ID NO: 566; Containing the VH sequence of SEQ ID NO: 451 and the VL sequence of SEQ ID NO: 567; Comprising the VH sequence of SEQ ID NO: 452 and the VL sequence of SEQ ID NO: 568; Containing the VH sequence of SEQ ID NO: 453 and the VL sequence of SEQ ID NO: 569; Containing the VH sequence of SEQ ID NO: 454 and the VL sequence of SEQ ID NO: 570; Containing the VH sequence of SEQ ID NO: 455 and the VL sequence of SEQ ID NO: 571; Containing the VH sequence of SEQ ID NO: 456 and the VL sequence of SEQ ID NO: 572; Containing the VH sequence of SEQ ID NO: 457 and the VL sequence of SEQ ID NO: 573; Containing the VH sequence of SEQ ID NO: 458 and the VL sequence of SEQ ID NO: 574; Containing the VH sequence of SEQ ID NO: 459 and the VL sequence of SEQ ID NO: 575; Containing the VH sequence of SEQ ID NO: 460 and the VL sequence of SEQ ID NO: 576; Comprising the VH sequence of SEQ ID NO: 461 and the VL sequence of SEQ ID NO: 577; Comprising the VH sequence of SEQ ID NO: 462 and the VL sequence of SEQ ID NO: 578; Containing the VH sequence of SEQ ID NO: 463 and the VL sequence of SEQ ID NO: 579; Containing the VH sequence of SEQ ID NO: 464 and the VL sequence of SEQ ID NO: 580; Containing the VH sequence of SEQ ID NO: 465 and the VL sequence of SEQ ID NO: 581; Comprising the VH sequence of SEQ ID NO: 466 and the VL sequence of SEQ ID NO: 582; Containing the VH sequence of SEQ ID NO: 467 and the VL sequence of SEQ ID NO: 583; Comprising the VH sequence of SEQ ID NO: 468 and the VL sequence of SEQ ID NO: 584; Containing the VH sequence of SEQ ID NO: 469 and the VL sequence of SEQ ID NO: 585; Containing the VH sequence of SEQ ID NO: 470 and the VL sequence of SEQ ID NO: 586; Comprising the VH sequence of SEQ ID NO: 471 and the VL sequence of SEQ ID NO: 587; Comprising the VH sequence of SEQ ID NO: 472 and the VL sequence of SEQ ID NO: 588; Containing the VH sequence of SEQ ID NO: 473 and the VL sequence of SEQ ID NO: 589; Containing the VH sequence of SEQ ID NO: 474 and the VL sequence of SEQ ID NO: 590; Containing the VH sequence of SEQ ID NO: 475 and the VL sequence of SEQ ID NO: 591; Containing the VH sequence of SEQ ID NO: 476 and the VL sequence of SEQ ID NO: 592; Containing the VH sequence of SEQ ID NO: 477 and the VL sequence of SEQ ID NO: 593; Containing the VH sequence of SEQ ID NO: 478 and the VL sequence of SEQ ID NO: 594; Containing the VH sequence of SEQ ID NO: 479 and the VL sequence of SEQ ID NO: 595; Containing the VH sequence of SEQ ID NO: 480 and the VL sequence of SEQ ID NO: 596; Containing the VH sequence of SEQ ID NO: 481 and the VL sequence of SEQ ID NO: 597; Comprising the VH sequence of SEQ ID NO: 482 and the VL sequence of SEQ ID NO: 598; Containing the VH sequence of SEQ ID NO: 483 and the VL sequence of SEQ ID NO: 599; Containing the VH sequence of SEQ ID NO: 484 and the VL sequence of SEQ ID NO: 600; Containing the VH sequence of SEQ ID NO: 485 and the VL sequence of SEQ ID NO: 601; Containing the VH sequence of SEQ ID NO: 486 and the VL sequence of SEQ ID NO: 602; Containing the VH sequence of SEQ ID NO: 487 and the VL sequence of SEQ ID NO: 603; Comprising the VH sequence of SEQ ID NO: 488 and the VL sequence of SEQ ID NO: 604; Containing the VH sequence of SEQ ID NO: 489 and the VL sequence of SEQ ID NO: 605; Containing the VH sequence of SEQ ID NO: 490 and the VL sequence of SEQ ID NO: 606; Containing the VH sequence of SEQ ID NO: 491 and the VL sequence of SEQ ID NO: 607; Containing the VH sequence of SEQ ID NO: 492 and the VL sequence of SEQ ID NO: 608; Containing the VH sequence of SEQ ID NO: 493 and the VL sequence of SEQ ID NO: 609; Containing the VH sequence of SEQ ID NO: 494 and the VL sequence of SEQ ID NO: 610; Comprising the VH sequence of SEQ ID NO: 495 and the VL sequence of SEQ ID NO: 611; Containing the VH sequence of SEQ ID NO: 496 and the VL sequence of SEQ ID NO: 612; Containing the VH sequence of SEQ ID NO: 497 and the VL sequence of SEQ ID NO: 613; Containing the VH sequence of SEQ ID NO: 498 and the VL sequence of SEQ ID NO: 614; Containing the VH sequence of SEQ ID NO: 499 and the VL sequence of SEQ ID NO: 615; Containing a VH sequence of SEQ ID NO: 500 and a VL sequence of SEQ ID NO: 616; Containing the VH sequence of SEQ ID NO: 501 and the VL sequence of SEQ ID NO: 617; Comprising the VH sequence of SEQ ID NO: 502 and the VL sequence of SEQ ID NO: 618; Containing the VH sequence of SEQ ID NO: 503 and the VL sequence of SEQ ID NO: 619; Containing the VH sequence of SEQ ID NO: 504 and the VL sequence of SEQ ID NO: 620; Comprising a VH sequence of SEQ ID NO: 505 and a VL sequence of SEQ ID NO: 621; Containing the VH sequence of SEQ ID NO: 506 and the VL sequence of SEQ ID NO: 622; Containing the VH sequence of SEQ ID NO: 507 and the VL sequence of SEQ ID NO: 623; Containing the VH sequence of SEQ ID NO: 508 and the VL sequence of SEQ ID NO: 624; Containing the VH sequence of SEQ ID NO: 509 and the VL sequence of SEQ ID NO: 625; Containing the VH sequence of SEQ ID NO: 510 and the VL sequence of SEQ ID NO: 626; Containing the VH sequence of SEQ ID NO: 511 and the VL sequence of SEQ ID NO: 627; Comprising the VH sequence of SEQ ID NO: 512 and the VL sequence of SEQ ID NO: 628; Containing the VH sequence of SEQ ID NO: 513 and the VL sequence of SEQ ID NO: 629; Comprising the VH sequence of SEQ ID NO: 514 and the VL sequence of SEQ ID NO: 630; Containing the VH sequence of SEQ ID NO: 515 and the VL sequence of SEQ ID NO: 631; Containing the VH sequence of SEQ ID NO: 516 and the VL sequence of SEQ ID NO: 632; Containing the VH sequence of SEQ ID NO: 517 and the VL sequence of SEQ ID NO: 633; Comprising the VH sequence of SEQ ID NO: 518 and the VL sequence of SEQ ID NO: 634; Containing the VH sequence of SEQ ID NO: 519 and the VL sequence of SEQ ID NO: 635; Containing the VH sequence of SEQ ID NO: 520 and the VL sequence of SEQ ID NO: 636; Containing the VH sequence of SEQ ID NO: 521 and the VL sequence of SEQ ID NO: 637; Comprising the VH sequence of SEQ ID NO: 522 and the VL sequence of SEQ ID NO: 638; Containing the VH sequence of SEQ ID NO: 523 and the VL sequence of SEQ ID NO: 639; Containing the VH sequence of SEQ ID NO: 524 and the VL sequence of SEQ ID NO: 640; Comprising the VH sequence of SEQ ID NO: 525 and the VL sequence of SEQ ID NO: 641; comprising the VH sequence of SEQ ID NO: 526 and the VL sequence of SEQ ID NO: 642; or An anti-MYCT1 human antibody or antigen-binding fragment thereof comprising the VH sequence of SEQ ID NO: 527 and the VL sequence of SEQ ID NO:

643.

7. In paragraph 1, An anti-MYCT1 human antibody or an antigen-binding fragment thereof, wherein the antigen-binding fragment comprises Fd, Fab, Fab', F(ab')2, dsFv, scFv and single domain antibody (sdAb).

8. A nucleic acid molecule encoding an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 7.

9. A recombinant expression vector comprising the nucleic acid molecule of clause 8.

10. A host cell transfected with the expression vector of clause 9.

11. An immunoconjugate comprising an antibody or antigen-binding fragment of any one of claims 1 to 7 and a cytotoxic agent.

12. A pharmaceutical composition for preventing or treating a MYCT1-related disease, comprising an antibody or antigen-binding fragment of any one of claims 1 to 7 as an active ingredient.

13. A pharmaceutical composition wherein the above MYCT1-related disease is an angiogenesis-related disease or cancer.

14. In paragraph 13, A pharmaceutical composition wherein the pharmaceutical composition is administered in combination with at least one additional therapeutic agent or therapeutic procedure.

15. In paragraph 14, A pharmaceutical composition wherein said at least one additional therapeutic agent or therapeutic procedure is selected from one or more of chemotherapy, targeted anti-cancer therapy, oncolytic drug, cytotoxic agent, immune-based therapy, cytokine, surgical procedure, radiation procedure, vaccine, or cell therapy.

16. A composition for diagnosing a MYCT1-related disease, comprising an antibody or antigen-binding fragment of any one of claims 1 to 7 as an active ingredient.

17. A pharmaceutical composition for enhancing sensitivity to an immunotherapy agent in cancer, comprising an antibody or antigen-binding fragment of any one of claims 1 to 7 as an active ingredient.

18. A chimeric antigen receptor (CAR) comprising an antigen binding domain that specifically binds to MYCT1, The above antigen binding domain is, A heavy chain CDR1 (HCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 4 to 43; A heavy chain CDR2 (HCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 44 to 85; A heavy chain CDR3 (HCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 86 to 187; A light chain CDR1 (LCDR-1) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 188 to 264; A light chain CDR2 (LCDR-2) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 265 to 320; and A chimeric antigen receptor comprising an antigen-binding fragment comprising a light chain CDR3 (LCDR-3) comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 321 to 411.

19. An immune effector cell expressing the chimeric antigen receptor of claim 18.

20. A method for treating a MYCT1-related disease, comprising administering to a subject a composition comprising a therapeutically effective amount of an antibody or antigen-binding fragment of any one of claims 1 to 7.

21. A method for enhancing the sensitivity of a subject to an immunotherapy agent, comprising administering to the subject a composition comprising a therapeutically effective amount of an antibody or antigen-binding fragment of any one of claims 1 to 7.

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