MAGE-A1 specific TCR and application thereof

Abstract

The present disclosure provides a MAGE-A1 specific TCR and uses thereof, and in particular relates to a TCR modified to improve binding affinity, a nucleic acid encoding the same, a vector comprising the nucleic acid of the invention, a cell comprising the TCR, the nucleic acid or the vector of the invention, and a method of preparing the cell of the invention. The disclosure also relates to conjugates or compositions comprising the TCRs of the invention, and methods of treating cancer by using the TCRs or cells of the invention.

Description

MAGE-A1-specific TCR and its uses

[0001] This international patent application claims the priority benefit of Chinese patent application CN202210622198.1 filed on June 1, 2022, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present disclosure relates to T cell receptors (TCRs) and their uses, in particular TCRs directed against MAGE-A1 and their uses in treating MAGE-A1 positive cancers. Background Art

[0003] TCR-T uses corresponding experimental techniques to clone TCRs that can specifically recognize specific HLA-tumor antigen peptide complexes, and then delivers the TCR gene coding sequence to more T cells through gene delivery methods such as lentivirus, thereby giving these T cells new antigen recognition specificity. Patient-derived T cells are transduced with TCR genes in vitro and amplified in large quantities, and can effectively recognize tumor cell-specific antigens. These T cells are returned to the patient and can specifically kill tumor cells and exert anti-tumor activity. Since both intracellular and extracellular antigens can be recognized by TCR after presentation via HLA, TCR-T can target most tumor-specific antigens, especially those that can recognize antigens within tumor cells (about 90% of all antigens). Therefore, TCR-T is expected to be used in the treatment of almost all tumors, especially various solid tumors.

[0004] Numerous studies have revealed that tumor cells express specific tumor-associated antigens on their surfaces, which are not expressed or expressed at low levels in normal tissue cells. Therefore, developing TCR-T products targeting these antigens is an important approach for tumor immunotherapy. Some of these antigens are specifically expressed on the surfaces of tumor cells and germ cells, known as "cancer-testis antigens." These antigen genes have naturally become prime targets for TCR-T product research. The melanoma-associated antigen gene family, MAGE, is the most thoroughly studied cancer-testis antigen to date, and MAGE-A1 was the first cancer-testis antigen gene to be identified. Numerous studies have demonstrated that MAGE-A1 is expressed on the cell surfaces of various solid tumors, including melanoma, lung cancer, colorectal cancer, cervical cancer, and breast cancer. Therefore, developing TCR-T products targeting MAGE-A1 may benefit a wider range of patients with these indications.

[0005] Several HLA-A0201-restricted TCRs targeting MAGE-A1 have been developed, namely “ht27” and “T1367” (Obenaus, M., C., Leisegang, M., Chen, Bruggen, P., A MAGE‐1 antigenic peptide recognized by human cytolytic T lymphocytes on HLA‐A2 tumor cells. Cancer Immunol. Immunother. 2005.54:1214–1220.). Among them, ht27 is a natural TCR sequence found in the human body, but the affinity of this TCR is low, making it difficult to effectively exert anti-tumor effects in the body; and "T1367" is a mouse-derived TCR sequence targeting the human MAGE-A1 antigen found in transgenic mice. This TCR has a higher affinity and can effectively kill tumor cells in in vitro experiments, but its application in human experiments may produce a strong autoimmune response, resulting in ineffective treatment.

[0006] Therefore, there is a need to develop TCR products with high affinity that specifically target MAGE-A1.

[0007] Summary of the Invention

[0008] The present invention uses the human TCR ht27 sequence as a template and uses in vitro mutagenesis to mutate the complementarity determining region (CDR) of ht27. High-affinity mutant clones are obtained by in vitro positive screening of the mutated TCRs. In vitro negative screening is then performed to exclude mutant clones with non-specific activation, thereby obtaining mutant TCR sequences with high affinity and no non-specific reactions. Furthermore, the specificity and non-specific reactions of the mutant TCR sequences are verified through in vitro experiments, ultimately obtaining mutant TCR sequences with strong specificity and high affinity.

[0009] Accordingly, in one aspect, the present disclosure provides a modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to the parent TCR:

[0010] (i) amino acid substitutions in the variable region of the β chain comprising

[0011] (a) amino acid substitution at one or more of the following positions in CDR2β: Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65, and / or

[0012] (b) amino acid substitutions at one or more of the following positions in CDR3β: Ser105, Glu108, Gln111, and Tyr114;

[0013] and / or

[0014] (ii) amino acid substitutions in the α chain variable region comprising

[0015] (c) amino acid substitution at one or more of the following positions in CDR1α: Ser29 and Ser36, and / or

[0016] (d) amino acid substitution at the following position in CDR2α: Asn59;

[0017] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0018] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0019] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(13) relative to the parent TCR:

[0020] (1) an amino acid substitution at position Asn57 selected from the group consisting of N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T;

[0021] (2) an amino acid substitution at position Glu58 selected from the group consisting of E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W;

[0022] (3) an amino acid substitution at position Gly59 selected from the group consisting of G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T;

[0023] (4) an amino acid substitution at position Ser63 selected from the group consisting of S63D, S63E, S63H, S63K, S63Q, S63R, and S63T;

[0024] (5) an amino acid substitution at position Lys64 selected from the group consisting of K64M, K64C, K64W, K64T, K64P, and K64A;

[0025] (6) an amino acid substitution at position Ala65 selected from the group consisting of A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y;

[0026] (7) an amino acid substitution at position Ser105 selected from the group consisting of S105L, S105N, S105H, S105Q, S105K, and S105R;

[0027] (8) an amino acid substitution at position Glu108 selected from the group consisting of E108D, E108F, E108T, E108I, E108K, and E108P;

[0028] (9) an amino acid substitution at position Gln111 selected from the group consisting of Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H;

[0029] (10) an amino acid substitution at position Tyr114 selected from the group consisting of Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S;

[0030] (11) an amino acid substitution at position Ser29 selected from the group consisting of S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N;

[0031] (12) an amino acid substitution at position Ser36 selected from the group consisting of S36M, S36L, S36Q, S36I, S36F, S36P, and S36V;

[0032] (13) An amino acid substitution at Asn59 selected from the group consisting of N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T.

[0033] In some embodiments, the modified TCR does not comprise a single S105N amino acid substitution relative to the parent TCR.

[0034] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR2β relative to the parent TCR: N57D, E58Y, G59Q, S63T, K64C and A65V; preferably, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V in CDR2β relative to the parent TCR.

[0035] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(3) in CDR3β relative to the parent TCR:

[0036] (1) Q111G or Q111S;

[0037] (2) Y114F, Y114L, or Y114W;

[0038] (3) S105L, S105Q, S105N or S105H;

[0039] Preferably, the modified TCR comprises the amino acid substitution Y114F, Y114W, Q111GY114F, Q111SY114F, S105L or S105N in CDR3β relative to the parent TCR.

[0040] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F, or Q111SY114F in CDR3β relative to the parent TCR.

[0041] In some embodiments, the modified TCR comprises the following amino acid substitutions in CDR1α relative to the parent TCR:

[0042] (1)S29L;

[0043] (2) S36M, S36L or S36Q.

[0044] In some embodiments, the modified TCR comprises the amino acid substitution N59F in CDR2α relative to the parent TCR.

[0045] In some embodiments, the modified TCR comprises:

[0046] (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and

[0047] (ii) a β chain variable region comprising

[0048] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0049] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0050] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0051] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0052] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0053] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0054] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0055] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0056] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0057] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0058] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0059] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0060] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0061] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0062] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0063] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively, or

[0064] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively.

[0065] In some embodiments, the modified TCR comprises:

[0066] (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and

[0067] (ii) an α chain variable region comprising

[0068] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0069] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0070] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0071] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, or

[0072] CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively.

[0073] In some embodiments, the modified TCR comprises:

[0074] (i) an α chain variable region comprising

[0075] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0076] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0077] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0078] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, or

[0079] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively; and

[0080] (ii) a β chain variable region comprising

[0081] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0082] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0083] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0084] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0085] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0086] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0087] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0088] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0089] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0090] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0091] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0092] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0093] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0094] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0095] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0096] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively, or

[0097] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively.

[0098] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence as set forth in SEQ ID NO:46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO:46, and a beta chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs:48-64, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs:48-64.

[0099] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 65-68 and 77, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 65-68 and 77, and a beta chain variable region having an amino acid sequence set forth in SEQ ID NO: 47, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 47.

[0100] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 65-68 and 77, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 65-68 and 77, and a beta chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 48-64, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 48-64.

[0101] In some embodiments, the modified TCR further comprises a TCR constant region. In some embodiments, the TCR constant region is a mouse constant region or a human constant region. In some embodiments, the TCR constant region includes a TCR α chain constant region and / or a TCR β chain constant region. In a preferred embodiment, the TCR α chain constant region and / or the TCR β chain constant region comprises at least one cysteine ​​substitution relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain. In a preferred embodiment, the TCR α chain constant region comprises an LVL substitution or an LIV substitution, such that the α chain constant region comprises the amino acid sequence LLVIVLRIL.

[0102] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence as shown in SEQ ID NO:69, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO:69, and a TCR beta chain having an amino acid sequence as shown in any one of SEQ ID NOs:3-19, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs:3-19.

[0103] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 20-23 and 78, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 20-23 and 78, and a TCR beta chain having an amino acid sequence set forth in SEQ ID NO: 70, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 70.

[0104] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 20-23 and 78, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 20-23 and 78, and a TCR beta chain having an amino acid sequence selected from any one of SEQ ID NOs: 3-19, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 3-19.

[0105] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

[0106] In some embodiments, the MHC molecule is of type HLA-A*02, such as type HLA-A*02:01 or type HLA-A*02:05.

[0107] In some embodiments, the modified TCR is soluble or membrane-bound.

[0108] In some embodiments, the modified TCR is isolated or purified.

[0109] In another aspect, the present disclosure provides a modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to the parent TCR:

[0110] (i) amino acid substitutions in the variable region of the β chain comprising

[0111] (a) amino acid substitution at one or more of the following positions in CDR1β: Gln29, Ala36, Thr37 and Thr38,

[0112] (b) amino acid substitutions at one or more of the following positions in CDR2β: Ser56, Asn57, Glu58, and Gly59, and / or

[0113] (c) amino acid substitutions at one or more of the following positions in CDR3β: Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115;

[0114] and / or

[0115] (ii) amino acid substitutions in the α chain variable region comprising

[0116] (d) amino acid substitution at one or more of the following positions in CDR1α: Ser28, Ser29, Ser36, Thr37 and Tyr38,

[0117] (e) amino acid substitutions in CDR2α at the following positions: Ile56, Phe57, Ser58, Asn59, Met63, and Asp64, and / or

[0118] (f) amino acid substitution at Thr26 in FR1α;

[0119] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0120] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0121] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(25) relative to the parent TCR:

[0122] (1) amino acid substitution Q29P at position Gln29 in CDR1β;

[0123] (2) amino acid substitution A36I at position Ala36 in CDR1β;

[0124] (3) amino acid substitution T37E at position Thr37 in CDR1β;

[0125] (4) amino acid substitution T38R at position Thr38 in CDR1β;

[0126] (5) amino acid substitution S56T at position Ser56 in CDR2β;

[0127] (6) an amino acid substitution at position Asn57 in CDR2β selected from the group consisting of N57Q and N57D;

[0128] (7) an amino acid substitution at position Glu58 in CDR2β selected from the group consisting of E58L, E58P, and E58Y;

[0129] (8) an amino acid substitution at position Gly59 in CDR2β selected from the group consisting of G59P and G59L;

[0130] (9) amino acid substitution of EPGQGP to VNDGT at positions Glu108 to Pro113 in CDR3β;

[0131] (10) amino acid substitution G112E at position Gly112 in CDR3β;

[0132] (11) amino acid substitution P113H at position Pro113 in CDR3β;

[0133] (12) amino acid substitution Y114L at position Tyr114 in CDR3β;

[0134] (13) amino acid substitution E115I at position Glu115 in CDR3β;

[0135] (14) an amino acid substitution at position Ser28 in CDR1α selected from the group consisting of S28Y and S28P;

[0136] (15) an amino acid substitution at position Ser29 in CDR1α selected from the group consisting of S29T, S29L, and S29Y;

[0137] (16) amino acid substitution S36D at position Ser36 in CDR1α;

[0138] (17) amino acid substitution T37I at position Thr37 in CDR1α;

[0139] (18) amino acid substitution Y38L at position Tyr38 in CDR1α;

[0140] (19) an amino acid substitution at position Ile56 in CDR2α selected from the group consisting of I56T and I56G;

[0141] (20) an amino acid substitution at position Phe57 in CDR2α selected from the group consisting of F57W and F57T;

[0142] (21) an amino acid substitution at position Ser58 in CDR2α selected from the group consisting of S58G, S58I, S58N, S58A, S58P, and S58W;

[0143] (22) an amino acid substitution at position Asn59 in CDR2α selected from the group consisting of N59F, N59T, N59H, and N59L;

[0144] (23) an amino acid substitution at position Met63 in CDR2α selected from the group consisting of M63D, M63Q, M63T, and M63E;

[0145] (24) an amino acid substitution at position Asp64 in CDR2α selected from the group consisting of D64T, D64Y, and D64S;

[0146] (25) An amino acid substitution at the Thr26 position in FR1α selected from the group consisting of T26A and T26V.

[0147] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR1β relative to the parent TCR: Q29P, A36I, T37E and T38R; preferably, the modified TCR comprises amino acid substitutions Q29P, A36I, T37E and T38R in CDR1β relative to the parent TCR.

[0148] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(4) in CDR2β relative to the parent TCR:

[0149] (1)S56T;

[0150] (2) N57Q or N57D;

[0151] (3) E58L, E58P, or E58Y;

[0152] (4) G59P or G59L;

[0153] Preferably, the modified TCR comprises the amino acid substitution S56TE58LG59P, S56TN57QE58PG59L or S56TN57DE58Y in CDR2β relative to the parent TCR.

[0154] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR3β relative to the parent TCR: G112E, P113H, Y114L, E115I, and substitution of EPGQGP at positions Glu108 to Pro113 to VNDGT; preferably, the modified TCR comprises amino acid substitutions in CDR3β relative to the parent TCR: substitution of EPGQGP at positions Glu108 to Pro113 to VNDGT or G112EP113HY114LE115I.

[0155] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR1α and / or FR1α relative to the parent TCR:

[0156] (1) S28Y or S28P;

[0157] (2) S29T, S29L, or S29Y;

[0158] (3)S36D;

[0159] (4)T37I;

[0160] (5)Y38L;

[0161] (6) T26A or T26V;

[0162] Preferably, the modified TCR comprises the following amino acid substitutions in CDR1α and / or FR1α relative to the parent TCR: S29TS36DT37IY38L, T26AS28YS29L or T26VS28PS29Y.

[0163] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR2α relative to the parent TCR:

[0164] (1)I56T or I56G;

[0165] (2) F57W or F57T;

[0166] (3) S58G, S58I, S58N, S58A, S58P or S58W;

[0167] (4) N59F, N59T, N59H or N59L;

[0168] (5) M63D, M63Q, M63T or M63E;

[0169] (6) D64T, D64Y or D64S;

[0170] Preferably, the modified TCR comprises the amino acid substitutions S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F in CDR2α relative to the parent TCR.

[0171] In some embodiments, the modified TCR comprises:

[0172] (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and

[0173] (ii) a β chain variable region comprising

[0174] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0175] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0176] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0177] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0178] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively, or

[0179] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively.

[0180] In some embodiments, the modified TCR comprises:

[0181] (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and

[0182] (ii) an α chain variable region comprising

[0183] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0184] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0185] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0186] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0187] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0188] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0189] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0190] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0191] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0192] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively, or

[0193] CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively.

[0194] In some embodiments, the modified TCR comprises:

[0195] (i) an α chain variable region comprising

[0196] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0197] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0198] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0199] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0200] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0201] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0202] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0203] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0204] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0205] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively, or

[0206] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively; and

[0207] (ii) a β chain variable region comprising

[0208] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0209] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0210] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0211] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0212] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively, or

[0213] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively.

[0214] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence as shown in SEQ ID NO: 46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 46, and a beta chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 80, 83, 86, 89, 92, 95, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 80, 83, 86, 89, 92, 95.

[0215] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and a beta chain variable region having an amino acid sequence set forth in SEQ ID NO: 47, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 47.

[0216] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and a beta chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 80, 83, 86, 89, 92, 95, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 80, 83, 86, 89, 92, 95.

[0217] In some embodiments, the modified TCR further comprises a TCR constant region. In some embodiments, the TCR constant region is a mouse constant region or a human constant region. In some embodiments, the TCR constant region includes a TCR α chain constant region and / or a TCR β chain constant region. In a preferred embodiment, the TCR α chain constant region and / or the TCR β chain constant region comprises at least one cysteine ​​substitution relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain. In a preferred embodiment, the TCR α chain constant region comprises an LVL substitution or an LIV substitution, such that the α chain constant region comprises the amino acid sequence LLVIVLRIL.

[0218] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence as shown in SEQ ID NO:69, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to SEQ ID NO:69, and a TCR beta chain having an amino acid sequence as shown in any one of SEQ ID NOs:81, 84, 87, 90, 93, 96, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to any one of SEQ ID NOs:81, 84, 87, 90, 93, 96.

[0219] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, and a TCR beta chain having an amino acid sequence set forth in SEQ ID NO: 70, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 70.

[0220] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, and a TCR beta chain having an amino acid sequence selected from any one of SEQ ID NOs: 81, 84, 87, 90, 93, 96, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 81, 84, 87, 90, 93, 96.

[0221] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

[0222] In some embodiments, the MHC molecule is of type HLA-A*02, such as type HLA-A*02:01 or type HLA-A*02:05.

[0223] In some embodiments, the modified TCR is soluble or membrane-bound.

[0224] In some embodiments, the modified TCR is isolated or purified.

[0225] In yet another aspect, the present disclosure provides a modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to the parent TCR:

[0226] (i) amino acid substitutions in the variable region of the β chain comprising

[0227] (a) amino acid substitution at one or more of the following positions in CDR1β: Gln29, Ala36, Thr37 and Thr38,

[0228] (b) amino acid substitutions at one or more of the following positions in CDR2β: Ser56, Asn57, Glu58, Ser63, and Lys64, and / or

[0229] (c) amino acid substitutions at one or more of the following positions in CDR3β: Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115;

[0230] and / or

[0231] (ii) amino acid substitutions in the α chain variable region comprising

[0232] (d) amino acid substitution at one or more of the following positions in CDR1α: Asp27, Ser28, Ser29 and Ser36,

[0233] (e) amino acid substitutions at the following positions in CDR2α: Ser58, Asn59, Met63 and Asp64, and / or

[0234] (f) amino acid substitution at Thr26 in FR1α;

[0235] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0236] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0237] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(24) relative to the parent TCR:

[0238] (1) an amino acid substitution at position Gln29 selected from the group consisting of Q29E and Q29P;

[0239] (2) amino acid substitution A36I at position Ala36;

[0240] (3) amino acid substitution T37E at Thr37;

[0241] (4) amino acid substitution T38R at Thr38 position;

[0242] (5) amino acid substitution S56T at position Ser56;

[0243] (6) amino acid substitution N57D at position Asn57;

[0244] (7) amino acid substitution E58Y at position Glu58;

[0245] (8) amino acid substitution S63D at position Ser63;

[0246] (9) amino acid substitution K64Q at Lys64;

[0247] (10) amino acid substitution G110W at position Gly110;

[0248] (11) amino acid substitution Q111D at position Gln111;

[0249] (12) an amino acid substitution at position Gly112 selected from the group consisting of G112P and G112A;

[0250] (13) amino acid substitution P113W at position Pro113;

[0251] (14) an amino acid substitution at position Tyr114 selected from the group consisting of Y114W, Y114S, and Y114F;

[0252] (15) amino acid substitution E115D at position Glu115;

[0253] (16) an amino acid substitution at position Asp27 selected from the group consisting of D27C and D27Y;

[0254] (17) an amino acid substitution at position Ser28 selected from the group consisting of S28P, S28M, S28A, and S28G;

[0255] (18) an amino acid substitution at position Ser29 selected from the group consisting of S29Y, S29L, S29F, S29E, and S29M;

[0256] (19) an amino acid substitution at position Ser36 selected from the group consisting of S36L, S36Q, S36M, and S36F;

[0257] (20) an amino acid substitution at position Ser58 selected from the group consisting of S58I and S58A;

[0258] (21) amino acid substitution N59F at position Asn59;

[0259] (22) an amino acid substitution at position Met63 selected from the group consisting of M63Q and M63E;

[0260] (23) an amino acid substitution at position Asp64 selected from the group consisting of D64A, D64S, and D64Q;

[0261] (24) An amino acid substitution at the Thr26 position selected from the group consisting of T26V, T26S, T26I, T26P and T26Q.

[0262] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(4) in CDR1β relative to the parent TCR:

[0263] (1) Q29E or Q29P;

[0264] (2)A36I;

[0265] (3)T37E;

[0266] (4)T38R;

[0267] Preferably, the modified TCR comprises the amino acid substitution Q29E or Q29PA36IT37ET38R in CDR1β relative to the parent TCR.

[0268] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR2β relative to the parent TCR: S56T, N57D, E58Y, S63D and K64Q; preferably, the modified TCR comprises the amino acid substitution S56TN57DE58Y or S63DK64Q in CDR2β relative to the parent TCR.

[0269] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR3β relative to the parent TCR:

[0270] (1)G110W;

[0271] (2)Q111D;

[0272] (3) G112P or G112A;

[0273] (4)P113W;

[0274] (5) Y114W, Y114S or Y114F;

[0275] (6)E115D;

[0276] Preferably, the modified TCR comprises the following amino acid substitutions in CDR3β relative to the parent TCR: G110WQ111DG112PP113W, Y114W, G112AP113WY114SE115D or Y114F.

[0277] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(3) relative to the parent TCR:

[0278] (1) amino acid substitutions Q29E or Q29PA36IT37ET38R in CDR1β;

[0279] (2) amino acid substitutions S56TN57DE58Y or S63DK64Q in CDR2β;

[0280] (3) Amino acid substitutions in CDR3β: G110WQ111DG112PP113W, Y114W, G112AP113WY114SE115D, or Y114F.

[0281] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(5) in CDR1α and / or FR1α relative to the parent TCR:

[0282] (1) D27C or D27Y;

[0283] (2) S28P, S28M, S28A or S28G;

[0284] (3) S29Y, S29L, S29F, S29E or S29M;

[0285] (4) S36L, S36Q, S36M or S36F;

[0286] (5) T26V, T26S, T26I, T26P or T26Q;

[0287] Preferably, the modified TCR comprises the following amino acid substitutions in CDR1α and / or FR1α relative to the parent TCR: T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F or T26QD27YS28GS29M.

[0288] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(4) in CDR2α relative to the parent TCR:

[0289] (1) S58I or S58A;

[0290] (2)N59F;

[0291] (3) M63Q or M63E;

[0292] (4) D64A, D64S or D64Q;

[0293] Preferably, the modified TCR comprises the amino acid substitutions N59F, S58IN59FM63QD64A, S58AN59FM63ED64S, S58AN59FD64Q or S58IN59FM63QD64A in CDR2α relative to the parent TCR.

[0294] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(2) relative to the parent TCR:

[0295] (1) amino acid substitutions in CDR1α and / or FR1α: T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F, or T26QD27YS28GS29M;

[0296] (2) Amino acid substitutions in CDR2α: N59F, S58IN59FM63QD64A, S58AN59FM63ED64S, S58AN59FD64Q or S58IN59FM63QD64A.

[0297] In some embodiments, the modified TCR comprises:

[0298] (i) an α chain variable region comprising

[0299] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively,

[0300] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively,

[0301] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively,

[0302] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively,

[0303] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively,

[0304] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively,

[0305] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively,

[0306] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0307] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively,

[0308] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or

[0309] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively,

[0310] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively,

[0311] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or

[0312] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively; and

[0313] (ii) a β chain variable region comprising

[0314] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively,

[0315] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively,

[0316] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively,

[0317] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively,

[0318] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively,

[0319] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or

[0320] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

[0321] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 132, 136, 142, 147, 150, 154, 158, 68, 166, 119, 168, 171, 173, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 132, 136, 142, 147, 150, 154, 158, 68, 166, 119, 168, 171, 173, and an amino acid sequence having an amino acid sequence selected from any one of SEQ ID NOs: 133, 137, 143, 155, 159, 163, 175, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: Any of NO: 133, 137, 143, 155, 159, 163, 175 has a beta chain variable region with an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

[0322] In some embodiments, the modified TCR further comprises a TCR constant region. In some embodiments, the TCR constant region is a mouse constant region or a human constant region. In some embodiments, the TCR constant region includes a TCR α chain constant region and / or a TCR β chain constant region. In a preferred embodiment, the TCR α chain constant region and / or the TCR β chain constant region comprises at least one cysteine ​​substitution relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain. In a preferred embodiment, the TCR α chain constant region comprises an LVL substitution or an LIV substitution, such that the α chain constant region comprises the amino acid sequence LLVIVLRIL.

[0323] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 134, 138, 144, 148, 151, 156, 160, 23, 167, 120, 169, 172, 174, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 134, 138, 144, 148, 151, 156, 160, 23, 167, 120, 169, 172, 174, and a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 135, 139, 145, 157, 161, 164, 176, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: Any of NO: 135, 139, 145, 157, 161, 164, 176 has a TCR beta chain with an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

[0324] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

[0325] In some embodiments, the MHC molecule is of type HLA-A*02, such as type HLA-A*02:01 or type HLA-A*02:05.

[0326] In some embodiments, the modified TCR is soluble or membrane-bound.

[0327] In some embodiments, the modified TCR is isolated or purified.

[0328] In another aspect, the present disclosure provides a modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to the parent TCR:

[0329] (i) amino acid substitutions in the variable region of the β chain comprising

[0330] (a) amino acid substitution at one or more of the following positions in CDR1β: Gln29, Ala36, Thr37 and Thr38,

[0331] (b) amino acid substitutions at one or more of the following positions in CDR2β: Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65, and / or

[0332] (c) amino acid substitutions at one or more of the following positions in CDR3β: Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115;

[0333] and / or

[0334] (ii) amino acid substitutions in the α chain variable region comprising

[0335] (d) amino acid substitution at one or more of the following positions in CDR1α: Ser28, Ser29, Ser36, Thr37 and Tyr38,

[0336] (e) amino acid substitutions in CDR2α at the following positions: Ile56, Phe57, Ser58, Asn59, Met63, and Asp64, and / or

[0337] (f) amino acid substitution at Thr26 in FR1α;

[0338] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0339] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0340] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(31) relative to the parent TCR:

[0341] (1) amino acid substitution Q29P at position Gln29 in CDR1β;

[0342] (2) amino acid substitution A36I at position Ala36 in CDR1β;

[0343] (3) amino acid substitution T37E at position Thr37 in CDR1β;

[0344] (4) amino acid substitution T38R at position Thr38 in CDR1β;

[0345] (5) amino acid substitution S56T at position Ser56 in CDR2β;

[0346] (6) an amino acid substitution at position Asn57 in CDR2β selected from the group consisting of N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T;

[0347] (7) an amino acid substitution at position Glu58 in CDR2β selected from the group consisting of E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W;

[0348] (8) an amino acid substitution at position Gly59 in CDR2β selected from the group consisting of G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T;

[0349] (9) an amino acid substitution at position Ser63 in CDR2β selected from the group consisting of S63D, S63E, S63H, S63K, S63Q, S63R, and S63T;

[0350] (10) an amino acid substitution at position Lys64 in CDR2β selected from the group consisting of K64M, K64C, K64W, K64T, K64P, and K64A;

[0351] (11) an amino acid substitution at position Ala65 in CDR2β selected from the group consisting of A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y;

[0352] (12) an amino acid substitution at position Ser105 in CDR3β selected from the group consisting of S105L, S105N, S105H, S105Q, S105K, and S105R;

[0353] (13) an amino acid substitution at position Glu108 in CDR3β selected from the group consisting of E108D, E108F, E108T, E108I, E108K, and E108P;

[0354] (14) an amino acid substitution at position Gln111 in CDR3β selected from the group consisting of Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H;

[0355] (15) amino acid substitution of EPGQGP to VNDGT at positions Glu108 to Pro113 in CDR3β;

[0356] (16) amino acid substitution G112E at position Gly112 in CDR3β;

[0357] (17) amino acid substitution P113H at position Pro113 in CDR3β;

[0358] (18) an amino acid substitution at position Tyr114 in CDR3β selected from the group consisting of Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S;

[0359] (19) amino acid substitution E115I at position Glu115 in CDR3β;

[0360] (20) an amino acid substitution at position Ser28 in CDR1α selected from the group consisting of S28Y and S28P;

[0361] (21) an amino acid substitution at position Ser29 in CDR1α selected from the group consisting of S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N;

[0362] (22) an amino acid substitution at position Ser36 in CDR1α selected from the group consisting of S36D, S36M, S36L, S36Q, S36I, S36F, S36P, and S36V;

[0363] (23) amino acid substitution T37I at position Thr37 in CDR1α;

[0364] (24) amino acid substitution Y38L at position Tyr38 in CDR1α;

[0365] (25) an amino acid substitution at position Ile56 in CDR2α selected from the group consisting of I56T and I56G;

[0366] (26) an amino acid substitution at position Phe57 in CDR2α selected from the group consisting of F57W and F57T;

[0367] (27) an amino acid substitution at position Ser58 in CDR2α selected from the group consisting of S58G, S58I, S58N, S58A, S58P, and S58W;

[0368] (28) an amino acid substitution at position Asn59 in CDR2α selected from the group consisting of N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T;

[0369] (29) an amino acid substitution at position Met63 in CDR2α selected from the group consisting of M63D, M63Q, M63T, and M63E;

[0370] (30) an amino acid substitution at position Asp64 in CDR2α selected from the group consisting of D64T, D64Y, and D64S;

[0371] (31) An amino acid substitution at position Thr26 in FR1α selected from the group consisting of T26A and T26V.

[0372] In some embodiments, the modified TCR does not comprise a single S105N amino acid substitution relative to the parent TCR.

[0373] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR1β relative to the parent TCR: Q29P, A36I, T37E and T38R; preferably, the modified TCR comprises amino acid substitutions Q29P, A36I, T37E and T38R in CDR1β relative to the parent TCR.

[0374] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(7) in CDR2β relative to the parent TCR:

[0375] (1)S56T;

[0376] (2) N57Q or N57D;

[0377] (3) E58L, E58P, or E58Y;

[0378] (4) G59Q, G59P, or G59L;

[0379] (5)S63T;

[0380] (6)K64C;

[0381] (7)A65V;

[0382] Preferably, the modified TCR comprises the amino acid substitution N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L or S56TN57DE58Y in CDR2β relative to the parent TCR.

[0383] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(7) in CDR3β relative to the parent TCR:

[0384] (1) Q111G or Q111S;

[0385] (2)G112E;

[0386] (3)P113H;

[0387] (4) Y114F, Y114L, or Y114W;

[0388] (5)E115I

[0389] (6) S105L, S105Q, S105N or S105H;

[0390] (7) EPGQGP at positions Glu108 to Pro113 was replaced by VNDGT;

[0391] Preferably, the modified TCR comprises the following amino acid substitutions in CDR3β relative to the parent TCR: Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, EPGQGP at positions Glu108 to Pro113 replaced by VNDGT or G112EP113HY114LE115I.

[0392] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F, or Q111SY114F in CDR3β relative to the parent TCR.

[0393] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR1α and / or FR1α relative to the parent TCR:

[0394] (1) S28Y or S28P;

[0395] (2) S29T, S29L, or S29Y;

[0396] (3) S36D, S36M, S36L or S36Q;

[0397] (4)T37I;

[0398] (5)Y38L;

[0399] (6) T26A or T26V;

[0400] Preferably, the modified TCR comprises the following amino acid substitutions in CDR1α and / or FR1α relative to the parent TCR: S29TS36DT37IY38L, T26AS28YS29L or T26VS28PS29Y.

[0401] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR2α relative to the parent TCR:

[0402] (1)I56T or I56G;

[0403] (2) F57W or F57T;

[0404] (3) S58G, S58I, S58N, S58A, S58P or S58W;

[0405] (4) N59F, N59T, N59H or N59L;

[0406] (5) M63D, M63Q, M63T or M63E;

[0407] (6) D64T, D64Y or D64S;

[0408] Preferably, the modified TCR comprises the amino acid substitutions S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F in CDR2α relative to the parent TCR.

[0409] In some embodiments, the modified TCR comprises:

[0410] (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and

[0411] (ii) a β chain variable region comprising

[0412] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0413] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0414] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0415] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0416] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0417] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0418] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0419] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0420] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0421] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0422] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0423] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0424] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0425] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0426] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0427] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively,

[0428] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively,

[0429] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0430] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0431] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0432] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0433] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively, or

[0434] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively.

[0435] In some embodiments, the modified TCR comprises:

[0436] (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and

[0437] (ii) an α chain variable region comprising

[0438] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0439] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0440] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0441] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0442] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively,

[0443] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0444] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0445] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0446] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0447] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0448] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0449] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0450] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0451] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0452] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively, or

[0453] CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively.

[0454] In some embodiments, the modified TCR comprises:

[0455] (i) an α chain variable region comprising

[0456] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0457] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0458] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0459] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0460] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively,

[0461] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0462] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0463] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0464] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0465] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0466] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0467] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0468] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0469] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0470] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively, or

[0471] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively; and

[0472] (ii) a β chain variable region comprising

[0473] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0474] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0475] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0476] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0477] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0478] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0479] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0480] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0481] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0482] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0483] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0484] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0485] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0486] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0487] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0488] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively,

[0489] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively,

[0490] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0491] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0492] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0493] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0494] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively, or

[0495] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively.

[0496] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence as set forth in SEQ ID NO: 46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 46, and a beta chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95.

[0497] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and a beta chain variable region having an amino acid sequence set forth in SEQ ID NO: 47, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 47.

[0498] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and an amino acid sequence selected from any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: Any of NOs: 48-64, 80, 83, 86, 89, 92, 95 has a beta chain variable region with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical.

[0499] In some embodiments, the modified TCR further comprises a TCR constant region. In some embodiments, the TCR constant region is a mouse constant region or a human constant region. In some embodiments, the TCR constant region includes a TCR α chain constant region and / or a TCR β chain constant region. In a preferred embodiment, the TCR α chain constant region and / or the TCR β chain constant region comprises at least one cysteine ​​substitution relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain. In a preferred embodiment, the TCR α chain constant region comprises an LVL substitution or an LIV substitution, such that the α chain constant region comprises the amino acid sequence LLVIVLRIL.

[0500] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence as shown in SEQ ID NO:69, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO:69, and a TCR beta chain having an amino acid sequence as shown in any one of SEQ ID NOs:3-19, 81, 84, 87, 90, 93, 96, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs:3-19, 81, 84, 87, 90, 93, 96.

[0501] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, and a TCR beta chain having an amino acid sequence set forth in SEQ ID NO: 70, or an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 70.

[0502] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, and an amino acid sequence having an amino acid sequence selected from any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: Any of NOs: 3-19, 81, 84, 87, 90, 93, 96 has a TCR beta chain with an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

[0503] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

[0504] In some embodiments, the MHC molecule is of type HLA-A*02, such as type HLA-A*02:01 or type HLA-A*02:05.

[0505] In some embodiments, the modified TCR is soluble or membrane-bound.

[0506] In some embodiments, the modified TCR is isolated or purified.

[0507] In yet another aspect, the present disclosure provides a modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to the parent TCR:

[0508] (i) amino acid substitutions in the variable region of the β chain comprising

[0509] (a) amino acid substitution at one or more of the following positions in CDR1β: Gln29, Ala36, Thr37 and Thr38,

[0510] (b) amino acid substitutions at one or more of the following positions in CDR2β: Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65, and / or

[0511] (c) amino acid substitutions at one or more of the following positions in CDR3β: Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115;

[0512] and / or

[0513] (ii) amino acid substitutions in the α chain variable region comprising

[0514] (d) amino acid substitution at one or more of the following positions in CDR1α: Asp27, Ser28, Ser29, Ser36, Thr37 and Tyr38,

[0515] (e) amino acid substitutions in CDR2α at the following positions: Ile56, Phe57, Ser58, Asn59, Met63, and Asp64, and / or

[0516] (f) amino acid substitution at Thr26 in FR1α;

[0517] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0518] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0519] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(33) relative to the parent TCR:

[0520] (1) an amino acid substitution at position Gln29 in CDR1β selected from the group consisting of Q29E and Q29P;

[0521] (2) amino acid substitution A36I at position Ala36 in CDR1β;

[0522] (3) amino acid substitution T37E at position Thr37 in CDR1β;

[0523] (4) amino acid substitution T38R at position Thr38 in CDR1β;

[0524] (5) amino acid substitution S56T at position Ser56 in CDR2β;

[0525] (6) an amino acid substitution at position Asn57 in CDR2β selected from the group consisting of N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T;

[0526] (7) an amino acid substitution at position Glu58 in CDR2β selected from the group consisting of E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W;

[0527] (8) an amino acid substitution at position Gly59 in CDR2β selected from the group consisting of G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T;

[0528] (9) an amino acid substitution at position Ser63 in CDR2β selected from the group consisting of S63D, S63E, S63H, S63K, S63Q, S63R, and S63T;

[0529] (10) an amino acid substitution at position Lys64 in CDR2β selected from the group consisting of K64Q, K64M, K64C, K64W, K64T, K64P, and K64A;

[0530] (11) an amino acid substitution at position Ala65 in CDR2β selected from the group consisting of A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y;

[0531] (12) an amino acid substitution at position Ser105 in CDR3β selected from the group consisting of S105L, S105N, S105H, S105Q, S105K, and S105R;

[0532] (13) an amino acid substitution at position Glu108 in CDR3β selected from the group consisting of E108D, E108F, E108T, E108I, E108K, and E108P;

[0533] (14) amino acid substitution G110W at position Gly110 in CDR3β;

[0534] (15) an amino acid substitution at position Gln111 in CDR3β selected from the group consisting of Q111D, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H;

[0535] (16) amino acid substitution of EPGQGP to VNDGT at positions Glu108 to Pro113 in CDR3β;

[0536] (17) an amino acid substitution at position Gly112 in CDR3β selected from the group consisting of G112P, G112A, and G112E;

[0537] (18) an amino acid substitution at position Pro113 in CDR3β selected from the group consisting of P113W and P113H;

[0538] (19) an amino acid substitution at position Tyr114 in CDR3β selected from the group consisting of Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S;

[0539] (20) an amino acid substitution at position Glu115 in CDR3β selected from the group consisting of E115D and E115I;

[0540] (21) an amino acid substitution at position Asp27 in CDR1α selected from the group consisting of D27C and D27Y;

[0541] (22) an amino acid substitution at position Ser28 in CDR1α selected from the group consisting of S28M, S28A, S28G, S28Y, and S28P;

[0542] (23) an amino acid substitution at position Ser29 in CDR1α selected from the group consisting of S29E, S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N;

[0543] (24) an amino acid substitution at position Ser36 in CDR1α selected from the group consisting of S36D, S36M, S36L, S36Q, S36I, S36F, S36P, and S36V;

[0544] (25) amino acid substitution T37I at position Thr37 in CDR1α;

[0545] (26) amino acid substitution Y38L at position Tyr38 in CDR1α;

[0546] (27) an amino acid substitution at position Ile56 in CDR2α selected from the group consisting of I56T and I56G;

[0547] (28) an amino acid substitution at position Phe57 in CDR2α selected from the group consisting of F57W and F57T;

[0548] (29) an amino acid substitution at position Ser58 in CDR2α selected from the group consisting of S58G, S58I, S58N, S58A, S58P, and S58W;

[0549] (30) an amino acid substitution at position Asn59 in CDR2α selected from the group consisting of N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T;

[0550] (31) an amino acid substitution at position Met63 in CDR2α selected from the group consisting of M63D, M63Q, M63T, and M63E;

[0551] (32) an amino acid substitution at position Asp64 in CDR2α selected from the group consisting of D64A, D64Q, D64T, D64Y, and D64S;

[0552] (33) An amino acid substitution at the Thr26 position in FR1α selected from the group consisting of T26S, T26I, T26P, T26Q, T26A, and T26V.

[0553] In some embodiments, the modified TCR does not comprise a single S105N amino acid substitution relative to the parent TCR.

[0554] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(4) in CDR1β relative to the parent TCR:

[0555] (1) Q29E or Q29P;

[0556] (2)A36I;

[0557] (3)T37E;

[0558] (4)T38R;

[0559] Preferably, the modified TCR comprises the amino acid substitution Q29E or Q29PA36IT37ET38R in CDR1β relative to the parent TCR.

[0560] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(7) in CDR2β relative to the parent TCR:

[0561] (1)S56T;

[0562] (2) N57Q or N57D;

[0563] (3) E58L, E58P, or E58Y;

[0564] (4) G59Q, G59P, or G59L;

[0565] (5) S63T or S63D;

[0566] (6) K64C or K64Q;

[0567] (7)A65V;

[0568] Preferably, the modified TCR comprises the amino acid substitution N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L, S56TN57DE58Y or S63DK64Q in CDR2β relative to the parent TCR.

[0569] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(8) in CDR3β relative to the parent TCR:

[0570] (1)G110W;

[0571] (2) Q111D, Q111G, or Q111S;

[0572] (3) G112E, G112P or G112A;

[0573] (4) P113H or P113W;

[0574] (5) Y114S, Y114F, Y114L or Y114W;

[0575] (6) E115I or E115D;

[0576] (7) S105L, S105Q, S105N or S105H;

[0577] (8) EPGQGP at positions Glu108 to Pro113 was substituted with VNDGT;

[0578] Preferably, the modified TCR comprises the following amino acid substitutions in CDR3β relative to the parent TCR: G110WQ111DG112PP113W, G112AP113WY114SE115D, Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, EPGQGP at positions Glu108 to Pro113 replaced by VNDGT or G112EP113HY114LE115I.

[0579] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(3) relative to the parent TCR:

[0580] (1) amino acid substitutions Q29E or Q29PA36IT37ET38R in CDR1β;

[0581] (2) amino acid substitutions in CDR2β: N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L, S56TN57DE58Y, or S63DK64Q;

[0582] (3) Amino acid substitutions in CDR3β: G110WQ111DG112PP113W, G112AP113WY114SE115D, Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, and EPGQGP at positions Glu108 to Pro113 were substituted with VNDGT or G112EP113HY114LE115I.

[0583] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F, or Q111SY114F in CDR3β relative to the parent TCR.

[0584] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(7) in CDR1α and / or FR1α relative to the parent TCR:

[0585] (1) D27C or D27Y;

[0586] (2) S28Y, S28P, S28M, S28A or S28G;

[0587] (3) S29T, S29L, S29Y, S29F, S29E or S29M;

[0588] (4) S36F, S36D, S36M, S36L or S36Q;

[0589] (5)T37I;

[0590] (6)Y38L;

[0591] (7) T26A, T26V, T26S, T26I, T26P or T26Q;

[0592] Preferably, the modified TCR comprises the following amino acid substitutions in CDR1α and / or FR1α relative to the parent TCR: S29TS36DT37IY38L, T26AS28YS29L, T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F or T26QD27YS28GS29M.

[0593] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR2α relative to the parent TCR:

[0594] (1)I56T or I56G;

[0595] (2) F57W or F57T;

[0596] (3) S58G, S58I, S58N, S58A, S58P or S58W;

[0597] (4) N59F, N59T, N59H or N59L;

[0598] (5) M63D, M63Q, M63T or M63E;

[0599] (6) D64A, D64Q, D64T, D64Y or D64S;

[0600] Preferably, the modified TCR comprises the amino acid substitutions N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F in CDR2α relative to the parent TCR.

[0601] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(2) relative to the parent TCR:

[0602] (1) CDR1α and / or FR1α contain the following amino acid substitutions: S29TS36DT37IY38L, T26AS28YS29L, T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F, or T26QD27YS28GS29M;

[0603] (2) CDR2α contains the amino acid substitutions N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L, or I56TS58WN59F.

[0604] In some embodiments, the modified TCR comprises:

[0605] (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and

[0606] (ii) a β chain variable region comprising

[0607] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0608] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0609] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0610] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0611] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0612] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0613] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0614] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0615] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0616] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0617] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0618] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0619] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0620] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0621] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0622] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively,

[0623] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively,

[0624] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0625] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0626] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0627] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0628] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively,

[0629] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively,

[0630] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively,

[0631] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively,

[0632] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively,

[0633] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively,

[0634] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively,

[0635] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or

[0636] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

[0637] In some embodiments, the modified TCR comprises:

[0638] (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and

[0639] (ii) an α chain variable region comprising

[0640] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0641] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0642] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0643] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0644] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively,

[0645] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0646] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0647] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0648] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0649] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0650] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0651] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0652] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0653] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0654] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively,

[0655] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively,

[0656] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively,

[0657] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively,

[0658] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively,

[0659] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively,

[0660] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively,

[0661] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively,

[0662] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively,

[0663] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0664] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively,

[0665] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or

[0666] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively,

[0667] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively,

[0668] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or

[0669] CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively.

[0670] In some embodiments, the modified TCR comprises:

[0671] (i) an α chain variable region comprising

[0672] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0673] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0674] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0675] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0676] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively,

[0677] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0678] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0679] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0680] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0681] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0682] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0683] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0684] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0685] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0686] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively,

[0687] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively,

[0688] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively,

[0689] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively,

[0690] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively,

[0691] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively,

[0692] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively,

[0693] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively,

[0694] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively,

[0695] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0696] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively,

[0697] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or

[0698] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively,

[0699] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively,

[0700] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or

[0701] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively; and

[0702] (ii) a β chain variable region comprising

[0703] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0704] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0705] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0706] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0707] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0708] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0709] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0710] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0711] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0712] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0713] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0714] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0715] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0716] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0717] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0718] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively,

[0719] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively,

[0720] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0721] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0722] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0723] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0724] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively,

[0725] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively,

[0726] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively,

[0727] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively,

[0728] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively,

[0729] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively,

[0730] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively,

[0731] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or

[0732] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

[0733] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence as set forth in SEQ ID NO: 46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 46, and a beta chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175.

[0734] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, or a sequence identical to or similar to SEQ ID NOs: NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 having an α chain variable region with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to any one of SEQ ID NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, and a β chain variable region having an amino acid sequence as set forth in SEQ ID NO:47, or an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to SEQ ID NO:47.

[0735] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, or a sequence identical to or similar to SEQ ID NOs: NO:98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity, and an α chain variable region having an amino acid sequence selected from any one of SEQ ID NO:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 or a sequence identical to SEQ ID NO: Any of NOs:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 has a beta chain variable region with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

[0736] In some embodiments, the modified TCR further comprises a TCR constant region. In some embodiments, the TCR constant region is a mouse constant region or a human constant region. In some embodiments, the TCR constant region includes a TCR α chain constant region and / or a TCR β chain constant region. In a preferred embodiment, the TCR α chain constant region and / or the TCR β chain constant region comprises at least one cysteine ​​substitution relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain. In a preferred embodiment, the TCR α chain constant region comprises an LVL substitution or an LIV substitution, such that the α chain constant region comprises the amino acid sequence LLVIVLRIL.

[0737] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence as set forth in SEQ ID NO: 69, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 69, and a TCR beta chain having an amino acid sequence as set forth in any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176.

[0738] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, or a sequence identical to or similar to SEQ ID NOs: NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 having a TCR alpha chain with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to any one of SEQ ID NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, and a TCR beta chain with an amino acid sequence shown in SEQ ID NO:70 or an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to SEQ ID NO:70.

[0739] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, or a sequence identical to or similar to SEQ ID NOs: NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 having an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity, and a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NO:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 or a sequence identical to SEQ ID NO: Any of NO:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 has a TCR beta chain with an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

[0740] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

[0741] In some embodiments, the MHC molecule is of type HLA-A*02, such as type HLA-A*02:01 or type HLA-A*02:05.

[0742] In some embodiments, the modified TCR is soluble or membrane-bound.

[0743] In some embodiments, the modified TCR is isolated or purified.

[0744] In another aspect, the disclosure provides nucleic acids encoding the modified TCRs of the invention.

[0745] In yet another aspect, the disclosure provides a vector comprising a nucleic acid of the invention.

[0746] In some embodiments, the vector is selected from the group consisting of a lentiviral vector, a retroviral vector, a plasmid, a DNA vector, an mRNA vector, a transposon-based vector, and an artificial chromosome.

[0747] In another aspect, the disclosure provides cells comprising a modified TCR, nucleic acid, or vector of the invention.

[0748] In some embodiments, the cell is selected from lymphocytes (e.g., T cells, NK cells), monocytes (e.g., PBMCs), and stem cells. In some embodiments, the stem cell is a lymphocyte progenitor cell or an induced pluripotent stem cell (iPSC). In some embodiments, the cell is a T cell. In some embodiments, the T cell does not express endogenous TCR.

[0749] In yet another aspect, the present disclosure provides a method for preparing the cell of the present invention, comprising the step of transducing or transfecting the cell with the vector of the present invention.

[0750] In some embodiments, the method further comprises the step of expanding and / or activating the cells before or after the transduction or transfection.

[0751] In another aspect, the present disclosure provides a conjugate comprising a modified TCR according to the present invention and an active agent coupled or conjugated to the TCR.

[0752] In some embodiments, the active agent is selected from a detectable label, an immunostimulatory molecule, and a therapeutic agent; preferably, the detectable label is selected from biotin, streptavidin, an enzyme or a catalytically active fragment thereof, a radionuclide, a nanoparticle, a paramagnetic metal ion, a nucleic acid probe, a contrast agent, and a fluorescent, phosphorescent or chemiluminescent molecule; preferably, the immunostimulatory molecule is selected from a cytokine, a chemokine, a platelet factor, and a complement initiator; preferably, the therapeutic agent is selected from an immunomodulator, a radioactive compound, an enzyme, a chemotherapeutic agent, and a toxin.

[0753] In yet another aspect, the present disclosure provides a bispecific or multispecific antigen-binding molecule comprising a modified TCR or antigen-binding fragment thereof of the present invention, and one or more additional antigen-binding regions that bind one or more additional antigens, optionally wherein the one or more additional antigen-binding regions are each independently selected from an antigen-binding region derived from an antibody and an antigen-binding region derived from a TCR.

[0754] In some embodiments, the one or more additional antigens are each independently selected from tumor-associated antigens (TAAs), tumor-specific antigens (TSAs), viral antigens, autoantigens, and immune cell surface molecules. In some embodiments, the one or more additional antigens are CD3, and preferably, the one or more additional antigen binding regions are derived from an antigen binding region of an anti-CD3 antibody.

[0755] In another aspect, the present disclosure provides a composition comprising a modified TCR, nucleic acid, vector, cell, conjugate, or bispecific or multispecific antigen-binding molecule according to the present invention; preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient.

[0756] In some embodiments, the composition further comprises a second therapeutic agent, preferably, the second therapeutic agent is selected from an antibody, a chemotherapeutic agent, and a small molecule drug.

[0757] In yet another aspect, the present disclosure provides a method of treating a MAGE-A1 positive cancer in a subject, comprising administering to the subject an effective amount of a modified TCR of the present invention.

[0758] In another aspect, the present disclosure provides a method of treating a MAGE-A1 positive cancer in a subject, comprising administering to the subject an effective amount of the cells of the present invention.

[0759] In some embodiments of the disclosed treatment methods, the cells are autologous or allogeneic to the subject.

[0760] In some embodiments, the method comprises the following steps:

[0761] (i) isolating a sample containing cells from the subject;

[0762] (ii) transducing or transfecting the cell with the vector of the present invention; and

[0763] (iii) administering the cells obtained in step (ii) to a subject.

[0764] In some embodiments, the method further comprises the step of knocking out the endogenous TCR in the cell after step (i) and before step (ii).

[0765] In some embodiments, the MAGE-A1 positive cancer is selected from melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial bladder carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[0766] In some embodiments, the method further comprises administering a second therapeutic agent, preferably, the second therapeutic agent is selected from an antibody, a chemotherapeutic agent, and a small molecule drug.

[0767] In some embodiments of the disclosed treatment methods, the subject has an HLA-A*02 allele, such as an HLA-A*02:01 or HLA-A*02:05 allele.

[0768] In another aspect, the disclosure provides use of a modified TCR, nucleic acid, vector, cell, conjugate, bispecific or multispecific antigen-binding molecule or composition of the invention in the preparation of a medicament for treating a MAGE-A1 positive cancer in a subject.

[0769] In yet another aspect, the disclosure provides a modified TCR, nucleic acid, vector, cell, conjugate, bispecific or multispecific antigen-binding molecule or composition of the invention for use in treating a MAGE-A1 positive cancer in a subject.

[0770] In some embodiments of the uses of the present disclosure, the MAGE-A1 positive cancer is selected from melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial carcinoma of the bladder), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma. In some embodiments, the subject has an HLA-A*02:01 allele, e.g., an HLA-A*02:01 or HLA-A*02:05 allele. BRIEF DESCRIPTION OF THE DRAWINGS

[0771] FIG1 shows the screening of affinity-optimized TCRs of the present invention by flow cytometry.

[0772] FIG2 shows the activation of target cells loaded with target antigen peptide (KVLEYVIKV) by affinity-optimized TCRs of the present invention as determined by flow cytometry.

[0773] FIG3 shows the activation of target cells loaded with gp100 target antigen by gp100 mutant TCRs known from the prior art by flow cytometry.

[0774] FIG4 shows the staining of cells expressing gp100 mutant TCRs known in the art with different concentrations of gp100 Dextramer as determined by flow cytometry.

[0775] FIG5 shows the results of gradient staining of cells expressing gp100 mutant TCRs known in the prior art using gp100 Dextramer.

[0776] FIG6 shows the linear relationship between Dextramer staining intensity and the affinity (KD-SPR) of gp100 mutant TCRs known in the prior art.

[0777] FIG. 7 shows the staining of cells expressing the mutant TCR of the present invention by different concentrations of MAGE-A1 Dextramer as determined by flow cytometry.

[0778] FIG8 shows the results of gradient staining of cells expressing the mutant TCR of the present invention using MAGE-A1 Dextramer.

[0779] FIG9 shows the phenotype of affinity-optimized TCR-T cells of the present invention determined by flow cytometry.

[0780] FIG10 shows the specific killing of antigen-positive tumor cells by affinity-optimized TCR-T cells of the present invention.

[0781] FIG11 shows the specific IFN-γ secretion of the affinity-optimized TCR-T cells of the present invention against antigen-positive tumor cells.

[0782] FIG12 shows the non-specific responses of affinity-optimized TCR-T cells of the present invention to different HLA typing, as determined by IFN-γ secretion experiments. DETAILED DESCRIPTION

[0783] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For example, the terms used herein are as in Janeway CA Jr, Travers P, Walport M et al., Immunobiology, 5th edition, New York: Garland Science (2001) and "A multilingual glossary of biotechnological terms: (IUPAC Recommendations)", Leuenberger, HGW, Nagel, B. and Definitions as described in H., ed. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland.

[0784] It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, the terms "a," "an," "one or more," and "at least one" can be used interchangeably. Similarly, the terms "comprising," "including," and "having" can be used interchangeably.

[0785] When the term "comprising" is used herein and in the appended claims, it does not exclude other elements. For the purposes of the present invention, the term "consisting of" is considered to be a preferred embodiment of the term "comprising". If a group is defined hereinafter as comprising or containing at least a certain number of embodiments, it should also be understood to disclose a group that preferably consists only of these embodiments.

[0786] In one aspect, the present disclosure provides a modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to a parent TCR:

[0787] (i) amino acid substitutions in the variable region of the β chain comprising

[0788] (a) amino acid substitution at one or more of the following positions in CDR1β: Gln29, Ala36, Thr37 and Thr38,

[0789] (b) amino acid substitutions at one or more of the following positions in CDR2β: Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65, and / or

[0790] (c) amino acid substitutions at one or more of the following positions in CDR3β: Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115;

[0791] and / or

[0792] (ii) amino acid substitutions in the α chain variable region comprising

[0793] (d) amino acid substitution at one or more of the following positions in CDR1α: Asp27, Ser28, Ser29, Ser36, Thr37 and Tyr38,

[0794] (e) amino acid substitutions in CDR2α at the following positions: Ile56, Phe57, Ser58, Asn59, Met63, and Asp64, and / or

[0795] (f) amino acid substitution at Thr26 in FR1α;

[0796] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0797] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0798] In some embodiments, the modified TCR comprises the following amino acid substitutions relative to the parent TCR:

[0799] (i) amino acid substitutions in the variable region of the β chain comprising

[0800] (a) amino acid substitution at one or more of the following positions in CDR2β: Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65, and / or

[0801] (b) amino acid substitutions at one or more of the following positions in CDR3β: Ser105, Glu108, Gln111, and Tyr114;

[0802] and / or

[0803] (ii) amino acid substitutions in the α chain variable region comprising

[0804] (c) amino acid substitution at one or more of the following positions in CDR1α: Ser29 and Ser36, and / or

[0805] (d) amino acid substitution at the following position in CDR2α: Asn59;

[0806] wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29;

[0807] The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

[0808] In its native form, the TCR exists as a complex of several proteins on the surface of the T cell. The T cell receptor consists of two (separate) protein chains, which are produced by independent T cell receptor α and β (TCRα and TCRβ) genes and are called α chain and β chain. Each chain of the TCR has an N-terminal immunoglobulin-like (Ig)-variable (V) region / domain, an Ig-constant (C) region / domain, a transmembrane / cell membrane spanning region that anchors the chain in the plasma membrane, and a short cytoplasmic tail at the C-terminus.

[0809] Typically, a TCR is a heterodimer of one α chain and one β chain that can bind to MHC molecules presenting antigenic peptides.

[0810] Antigen specificity is conferred by the variable regions of the α and β chains. Both variable regions of the TCR α and β chains (the α chain variable region (Vα) and the β chain variable region (Vβ)) contain three hypervariable or complementarity-determining regions (CDR1α / β, CDR2α / β, and CDR3α / β) surrounded by four framework (FR) regions (FR1α / β, FR2α / β, FR3α / β, and FR4α / β). CDR3 is the primary determinant of antigen recognition and specificity (i.e., the ability to recognize and interact with a specific antigen), while CDR1 and CDR2 primarily interact with the MHC molecules that present the antigenic peptide.

[0811] TCR recognizes antigenic peptides that are bound to major histocompatibility complex (MHC) molecules at the surface of antigen-presenting cells ("presented / displayed on MHC molecules"). Antigenic peptides presented on MHC molecules are also referred to herein as "complexes of epitopes and MHC molecules," "epitope-MHC complexes," or "target antigen peptide-MHC complexes." There are two different classes of MHC molecules: MHC I and MHC II, which present peptides from different cell compartments. MHC class I molecules are expressed on the surface of all nucleated cells in the human body and display peptides or protein fragments from intracellular compartments to cytotoxic T cells. In humans, MHC is also known as human leukocyte antigen (HLA). There are three main types of MHC class I: HLA-A, HLA-B, and HLA-C. Once the TCR binds to its specific epitope-MHC complex, the T cell is activated and exerts biological effector functions.

[0812] The positions of the amino acid sequences of TCRs in the present invention are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT). For example, the position numbering of the amino acid sequences of the TCR α chain variable region (Vα) and the β chain variable region (Vβ) are numbered according to the positions listed in IMGT. For example, if an amino acid in Vα is numbered 29 in IMGT, it will be described as the 29th amino acid in Vα in the present invention; if an amino acid in Vβ is numbered 57 in IMGT, it will be described as the 57th amino acid in Vβ in the present invention; and so on. In this article, if there are special instructions for the sequence position numbering of other amino acids, the special instructions shall apply.

[0813] The term "parent TCR" as used herein refers to the previously developed HLA-A0201-restricted human TCR ht27 targeting MAGE-A1, which comprises a CDR1β as shown in the amino acid sequence of SEQ ID NO: 24 (DFQATT), a CDR2β as shown in the amino acid sequence of SEQ ID NO: 25 (SNEGSKA), a CDR3β as shown in the amino acid sequence of SEQ ID NO: 26 (SAREPGQGPYEQY), a CDR1α as shown in the amino acid sequence of SEQ ID NO: 27 (DSSSTY), a CDR2α as shown in the amino acid sequence of SEQ ID NO: 28 (IFSNMDM), and a CDR3α as shown in the amino acid sequence of SEQ ID NO: 29 (AGSGGGTDKLI), and the amino acid sequences of its α chain variable region and β chain variable region are shown in SEQ ID NO: 46 and SEQ ID NO: 47, respectively.

[0814] α chain variable region:

[0815] β chain variable region:

[0816] The present inventors have discovered that modified TCRs of the present invention comprising amino acid substitutions at one or more of the following variable region positions relative to the parent TCR have improved binding affinity for MAGE-A1: Asn57, Glu58, Gly59, Ser63, Lys64, Ala65 in CDR2β; Ser105, Glu108, Gln111, Tyr114 in CDR3β; Ser29 and Ser36 in CDR1α; and Asn59 in CDR2α. For example, an amino acid substitution at Ser105 can increase the binding affinity of the TCR by approximately 10-400 fold; an amino acid substitution at Glu108 can increase the binding affinity of the TCR by approximately 1-2 fold; an amino acid substitution at Gln111 can increase the binding affinity of the TCR by approximately 12-14 fold; and an amino acid substitution at Tyr114 can increase the binding affinity of the TCR by approximately 11-944 fold. Specific amino acid substitutions at these amino acid positions include, but are not limited to, N57D, N57E, N57H, N57K, N57Q, N57R, N57S, N57T, E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, E58W, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, 59S, G59T, S63D, S63E, S63H, S63K, S63Q, S63R, S63T, K64M, K64C, K64W, K64T, K64P, K 64A, A65F, A65I, A65L, A65M, A65P, A65V, A65W, A65Y, S105L, S105N, S105H, S105Q, S1 05K, S105R, E108D, E108F, E108T, E108I, E108K, E108P, Q111N, Q111K, Q111S, Q111G , Q111A, Q111V, Q111E, Q111L, Q111H, Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y11 4N, Y114P, Y114S, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, S29N, S36M, S36L, S36Q, S36I, S36F, S36P, S36V, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T.

[0817] Thus, in some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(33) relative to the parent TCR:

[0818] (1) an amino acid substitution at position Gln29 in CDR1β selected from the group consisting of Q29E and Q29P;

[0819] (2) amino acid substitution A36I at position Ala36 in CDR1β;

[0820] (3) amino acid substitution T37E at position Thr37 in CDR1β;

[0821] (4) amino acid substitution T38R at position Thr38 in CDR1β;

[0822] (5) amino acid substitution S56T at position Ser56 in CDR2β;

[0823] (6) an amino acid substitution at position Asn57 in CDR2β selected from the group consisting of N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T;

[0824] (7) an amino acid substitution at position Glu58 in CDR2β selected from the group consisting of E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W;

[0825] (8) an amino acid substitution at position Gly59 in CDR2β selected from the group consisting of G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T;

[0826] (9) an amino acid substitution at position Ser63 in CDR2β selected from the group consisting of S63D, S63E, S63H, S63K, S63Q, S63R, and S63T;

[0827] (10) an amino acid substitution at position Lys64 in CDR2β selected from the group consisting of K64Q, K64M, K64C, K64W, K64T, K64P, and K64A;

[0828] (11) an amino acid substitution at position Ala65 in CDR2β selected from the group consisting of A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y;

[0829] (12) an amino acid substitution at position Ser105 in CDR3β selected from the group consisting of S105L, S105N, S105H, S105Q, S105K, and S105R;

[0830] (13) an amino acid substitution at position Glu108 in CDR3β selected from the group consisting of E108D, E108F, E108T, E108I, E108K, and E108P;

[0831] (14) amino acid substitution G110W at position Gly110 in CDR3β;

[0832] (15) an amino acid substitution at position Gln111 in CDR3β selected from the group consisting of Q111D, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H;

[0833] (16) amino acid substitution of EPGQGP to VNDGT at positions Glu108 to Pro113 in CDR3β;

[0834] (17) an amino acid substitution at position Gly112 in CDR3β selected from the group consisting of G112P, G112A, and G112E;

[0835] (18) an amino acid substitution at position Pro113 in CDR3β selected from the group consisting of P113W and P113H;

[0836] (19) an amino acid substitution at position Tyr114 in CDR3β selected from the group consisting of Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S;

[0837] (20) an amino acid substitution at position Glu115 in CDR3β selected from the group consisting of E115D and E115I;

[0838] (21) an amino acid substitution at position Asp27 in CDR1α selected from the group consisting of D27C and D27Y;

[0839] (22) an amino acid substitution at position Ser28 in CDR1α selected from the group consisting of S28M, S28A, S28G, S28Y, and S28P;

[0840] (23) an amino acid substitution at position Ser29 in CDR1α selected from the group consisting of S29E, S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N;

[0841] (24) an amino acid substitution at position Ser36 in CDR1α selected from the group consisting of S36D, S36M, S36L, S36Q, S36I, S36F, S36P, and S36V;

[0842] (25) amino acid substitution T37I at position Thr37 in CDR1α;

[0843] (26) amino acid substitution Y38L at position Tyr38 in CDR1α;

[0844] (27) an amino acid substitution at position Ile56 in CDR2α selected from the group consisting of I56T and I56G;

[0845] (28) an amino acid substitution at position Phe57 in CDR2α selected from the group consisting of F57W and F57T;

[0846] (29) an amino acid substitution at position Ser58 in CDR2α selected from the group consisting of S58G, S58I, S58N, S58A, S58P, and S58W;

[0847] (30) an amino acid substitution at position Asn59 in CDR2α selected from the group consisting of N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T;

[0848] (31) an amino acid substitution at position Met63 in CDR2α selected from the group consisting of M63D, M63Q, M63T, and M63E;

[0849] (32) an amino acid substitution at position Asp64 in CDR2α selected from the group consisting of D64A, D64Q, D64T, D64Y, and D64S;

[0850] (33) An amino acid substitution at the Thr26 position in FR1α selected from the group consisting of T26S, T26I, T26P, T26Q, T26A, and T26V.

[0851] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(13) relative to the parent TCR:

[0852] (1) an amino acid substitution at position Asn57 selected from the group consisting of N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T;

[0853] (2) an amino acid substitution at position Glu58 selected from the group consisting of E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W;

[0854] (3) an amino acid substitution at position Gly59 selected from the group consisting of G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T;

[0855] (4) an amino acid substitution at position Ser63 selected from the group consisting of S63D, S63E, S63H, S63K, S63Q, S63R, and S63T;

[0856] (5) an amino acid substitution at position Lys64 selected from the group consisting of K64M, K64C, K64W, K64T, K64P, and K64A;

[0857] (6) an amino acid substitution at position A1A65 selected from the group consisting of A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y;

[0858] (7) an amino acid substitution at position Ser105 selected from the group consisting of S105L, S105N, S105H, S105Q, S105K, and S105R;

[0859] (8) an amino acid substitution at position Glu108 selected from the group consisting of E108D, E108F, E108T, E108I, E108K, and E108P;

[0860] (9) an amino acid substitution at position Gln111 selected from the group consisting of Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H;

[0861] (10) an amino acid substitution at position Tyr114 selected from the group consisting of Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S;

[0862] (11) an amino acid substitution at position Ser29 selected from the group consisting of S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N;

[0863] (12) an amino acid substitution at position Ser36 selected from the group consisting of S36M, S36L, S36Q, S36I, S36F, S36P, and S36V;

[0864] (13) An amino acid substitution at Asn59 selected from the group consisting of N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T.

[0865] In some embodiments, the modified TCR does not comprise a single S105N amino acid substitution relative to the parent TCR. In some embodiments, the modified TCR comprises a combination of the amino acid substitution S105N and other amino acid substitutions listed above relative to the parent TCR.

[0866] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR2β relative to the parent TCR: N57D, E58Y, G59Q, S63T, K64C, and A65V. In a preferred embodiment, the modified TCR comprises amino acid substitutions in CDR2β relative to the parent TCR: N57D, E58Y, G59Q, S63T, K64C, and A65V.

[0867] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(3) relative to the parent TCR in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H. In some embodiments, the modified TCR comprises amino acid substitutions Q111GY114F, Q111GY114L, Q111GY114W, Q111SY114F, Q1114SY114L, or Q114SY114W relative to the parent TCR in CDR3β. In preferred embodiments, the modified TCR comprises the amino acid substitution Y114F, Y114W, Q111GY114F, Q111SY114F, S105L or S105N in CDR3β relative to the parent TCR.

[0868] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR2β relative to the parent TCR: N57D, E58Y, G59Q, S63T, K64C, and A65V; and one or more of the following amino acid substitutions (1)-(3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H.

[0869] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β relative to the parent TCR; and comprises one or more of the following amino acid substitutions (1)-(3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H.

[0870] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and amino acid substitutions Q111GY114F, Q111GY114L, Q111GY114W, Q111SY114F, Q1114SY114L, or Q114SY114W in CDR3β relative to the parent TCR.

[0871] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and amino acid substitutions Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, or S105N in CDR3β relative to the parent TCR.

[0872] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F, or Q111SY114F in CDR3β relative to the parent TCR.

[0873] In some embodiments, the modified TCR comprises an amino acid substitution in CDR1α of the parent TCR: (1) S29L; (2) S36M, S36L, or S36Q.

[0874] In some embodiments, the modified TCR comprises the amino acid substitution N59F in CDR2α relative to the parent TCR.

[0875] In some embodiments, the modified TCR comprises the following amino acid substitutions (1) and / or (2) in CDR1α relative to the parent TCR: (1) S29L; (2) S36M, S36L or S36Q; and comprises the amino acid substitution N59F in CDR2α.

[0876] The three CDR regions of each TCR α and β chain bind to different sites on the MHC-peptide complex, interacting with the complex in a relatively independent manner. The CDR mutations disclosed herein, whether alone or in combination, effectively increase TCR affinity relative to wild-type TCRs. Some combinations of mutations can even produce an additive effect, achieving even higher affinity.

[0877] Thus, in some embodiments, the modified TCR comprises, relative to the parent TCR, an amino acid substitution in CDR1α of (1) and / or (2): (1) S29L; (2) S36M, S36L, or S36Q; an amino acid substitution N59F in CDR2α; and one or more of the following amino acid substitutions in CDR2β: N57D, E58Y, G59Q, S63T, K64C, and A65V; and one or more of the following amino acid substitutions (1)-(3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H.

[0878] In some embodiments, the modified TCR comprises the following amino acid substitutions (1) and / or (2) in CDR1α relative to the parent TCR: (1) S29L; (2) S36M, S36L, or S36Q; the amino acid substitution N59F in CDR2α; and the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β; and one or more of the following amino acid substitutions (1)-(3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H.

[0879] In some embodiments, the modified TCR comprises the following amino acid substitutions (1) and / or (2) relative to the parent TCR in CDR1α: (1) S29L; (2) S36M, S36L, or S36Q; in CDR2α, the amino acid substitution N59F; and in CDR2β, the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V; and in CDR3β, the amino acid substitutions Q111GY114F, Q111GY114L, Q111GY114W, Q111SY114F, Q1114SY114L, or Q114SY114W.

[0880] In some embodiments, the modified TCR comprises the following amino acid substitutions (1) and / or (2) relative to the parent TCR in CDR1α: (1) S29L; (2) S36M, S36L or S36Q; in CDR2α, the amino acid substitution N59F; and in CDR2β, the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V; and in CDR3β, the amino acid substitutions Y114F, Y114W, Q111GY114F, Q111SY114F, S105L or S105N.

[0881] In some embodiments, the modified TCR comprises the following amino acid substitutions (1) and / or (2) relative to the parent TCR in CDR1α: (1) S29L; (2) S36M, S36L or S36Q; in CDR2α, the amino acid substitution N59F; and in CDR2β, the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V; and in CDR3β, the amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F or Q111SY114F.

[0882] In some embodiments, the modified TCR comprises:

[0883] (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and

[0884] (ii) a β chain variable region comprising

[0885] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0886] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0887] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0888] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0889] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0890] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0891] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0892] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0893] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0894] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0895] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0896] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0897] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0898] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0899] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0900] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively,

[0901] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively,

[0902] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[0903] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[0904] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[0905] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[0906] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively,

[0907] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively,

[0908] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively,

[0909] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively,

[0910] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively,

[0911] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively,

[0912] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively,

[0913] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or

[0914] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

[0915] In some embodiments, the modified TCR comprises:

[0916] (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and

[0917] (ii) a β chain variable region comprising

[0918] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[0919] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[0920] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[0921] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[0922] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[0923] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[0924] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[0925] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[0926] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[0927] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[0928] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[0929] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[0930] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[0931] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[0932] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[0933] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively, or

[0934] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively.

[0935] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 30 and 26, respectively.

[0936] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 31, respectively.

[0937] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 32, respectively.

[0938] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 33, respectively.

[0939] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 34, respectively.

[0940] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 35, respectively.

[0941] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 30 and 33, respectively.

[0942] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 30 and 34, respectively.

[0943] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 30 and 35, respectively.

[0944] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 36, respectively.

[0945] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 37, respectively.

[0946] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 30 and 36, respectively.

[0947] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 30 and 37, respectively.

[0948] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 38, respectively.

[0949] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 39, respectively.

[0950] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 40, respectively.

[0951] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 41, respectively.

[0952] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 79, respectively.

[0953] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 82, 25 and 26, respectively.

[0954] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 85 and 26, respectively.

[0955] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 88 and 26, respectively.

[0956] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 91 and 26, respectively.

[0957] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24, 25 and 94, respectively.

[0958] In some embodiments, the modified TCR comprises:

[0959] (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and

[0960] (ii) an α chain variable region comprising

[0961] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0962] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0963] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0964] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0965] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively,

[0966] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[0967] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[0968] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[0969] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[0970] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[0971] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[0972] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[0973] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[0974] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[0975] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively,

[0976] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively,

[0977] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively,

[0978] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively,

[0979] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively,

[0980] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively,

[0981] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively,

[0982] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively,

[0983] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively,

[0984] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[0985] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively,

[0986] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or

[0987] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively,

[0988] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively,

[0989] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or

[0990] CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively.

[0991] In some embodiments, the modified TCR comprises:

[0992] (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and

[0993] (ii) an α chain variable region comprising

[0994] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[0995] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[0996] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[0997] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, or

[0998] CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively.

[0999] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 42, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1000] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 43, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1001] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 44, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1002] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 45, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1003] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 76 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1004] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 97, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1005] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 100, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1006] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 103, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1007] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 106 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1008] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 109 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1009] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 112 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1010] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 115 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1011] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 118 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1012] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 121 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1013] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 124 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1014] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 127 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 24-26, respectively.

[1015] In some embodiments, the modified TCR comprises:

[1016] (i) an α chain variable region comprising

[1017] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[1018] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[1019] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[1020] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[1021] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively,

[1022] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively,

[1023] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively,

[1024] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively,

[1025] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively,

[1026] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively,

[1027] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively,

[1028] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively,

[1029] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively,

[1030] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively,

[1031] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively,

[1032] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively,

[1033] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively,

[1034] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively,

[1035] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively,

[1036] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively,

[1037] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively,

[1038] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively,

[1039] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively,

[1040] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively,

[1041] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively,

[1042] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or

[1043] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively,

[1044] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively,

[1045] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or

[1046] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively; and

[1047] (ii) a β chain variable region comprising

[1048] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[1049] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[1050] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[1051] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[1052] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[1053] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[1054] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[1055] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[1056] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[1057] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[1058] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[1059] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[1060] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[1061] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[1062] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[1063] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively,

[1064] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively,

[1065] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively,

[1066] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively,

[1067] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively,

[1068] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively,

[1069] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively,

[1070] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively,

[1071] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively,

[1072] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively,

[1073] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively,

[1074] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively,

[1075] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively,

[1076] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or

[1077] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

[1078] In some embodiments, the modified TCR comprises:

[1079] (i) an α chain variable region comprising

[1080] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively,

[1081] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively,

[1082] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively,

[1083] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, or

[1084] CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively; and

[1085] (ii) a β chain variable region comprising

[1086] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively,

[1087] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively,

[1088] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively,

[1089] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively,

[1090] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively,

[1091] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively,

[1092] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively,

[1093] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively,

[1094] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively,

[1095] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively,

[1096] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively,

[1097] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively,

[1098] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively,

[1099] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively,

[1100] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively,

[1101] CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively, or

[1102] CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively.

[1103] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 103, 76 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 25 and 131, respectively.

[1104] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively.

[1105] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 140, 141 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1106] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 146, 118 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1107] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 45, 149 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1108] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 152, 149 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 153 and 35, respectively.

[1109] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively.

[1110] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 45, 28 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 162, respectively.

[1111] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 165, 118 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1112] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 27, 118 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1113] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 45, 141 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1114] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1115] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 43, 118 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 130, 91 and 35, respectively.

[1116] In a preferred embodiment, the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α having amino acid sequences as shown in SEQ ID NOs: 152, 149 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β having amino acid sequences as shown in SEQ ID NOs: 82, 91 and 33, respectively.

[1117] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence as set forth in SEQ ID NO: 46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 46; and / or an amino acid sequence selected from any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, or 183, or having a sequence identical to SEQ ID NO: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, or 183, or having a sequence identical to SEQ ID NO: Any of NOs:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 has a beta chain variable region of an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[1118] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, or a sequence identical to or similar to SEQ ID NOs: NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 having an alpha chain variable region having an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to any one of SEQ ID NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173; and / or a beta chain variable region having an amino acid sequence as set forth in SEQ ID NO:47, or an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to SEQ ID NO:47.

[1119] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, or a sequence identical to or similar to SEQ ID NOs: NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity; and / or having an amino acid sequence selected from any one of SEQ ID NO:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 or having an amino acid sequence identical to any one of SEQ ID NO: Any of NOs:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 has a beta chain variable region with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

[1120] As used herein, the term "sequence identity" refers to the extent to which two (nucleotide or amino acid) sequences have the same residue at the same position in an alignment, and is typically expressed as a percentage. Preferably, identity is determined over the entire length of the sequences being compared. Thus, two copies of exactly the same sequence have 100% identity, but sequences that are less highly conserved and have deletions, additions, or substitutions may have a lower degree of identity. Those skilled in the art will recognize that several algorithms can be used to determine sequence identity using standard parameters, such as Blast (Altschul et al. (1997) Nucleic Acids Res. 25: 3389-3402), Blast2 (Altschul et al. (1990) J. Mol. Biol. 215: 403-410), Smith-Waterman (Smith et al. (1981) J. Mol. Biol. 147: 195-197), and ClustalW.

[1121] Thus, for example, the amino acid sequence of SEQ ID NO: 46 or 47 can be used as a "subject sequence" or "reference sequence," while a TCR α chain or β chain variable region amino acid sequence that differs therefrom can be used as a "query sequence."

[1122] In some embodiments, the modified TCR comprises an α chain variable region having an amino acid sequence as shown in SEQ ID NO: 46; and a β chain variable region having an amino acid sequence as shown in any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175.

[1123] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173; and a beta chain variable region having an amino acid sequence shown in SEQ ID NO: 47.

[1124] In some embodiments, the modified TCR comprises an alpha chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173; and a beta chain variable region having an amino acid sequence selected from any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175.

[1125] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:48.

[1126] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:49.

[1127] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:50.

[1128] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:51.

[1129] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:52.

[1130] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:53.

[1131] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:54.

[1132] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:55.

[1133] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:56.

[1134] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:57.

[1135] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:58.

[1136] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:59.

[1137] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:60.

[1138] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:61.

[1139] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:62.

[1140] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:63.

[1141] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:64.

[1142] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 65, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1143] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 66, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1144] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 67, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1145] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 68, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1146] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 77, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1147] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:80.

[1148] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:83.

[1149] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:86.

[1150] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:89.

[1151] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:92.

[1152] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO:46, and a β chain variable region having the amino acid sequence shown in SEQ ID NO:95.

[1153] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 98, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1154] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 101, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1155] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 104, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1156] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 107, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1157] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 110, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1158] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 113, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1159] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 116, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1160] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 119, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1161] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 122, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1162] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 125, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1163] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 128, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 47.

[1164] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 132, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 133.

[1165] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 136, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 137.

[1166] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 142, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1167] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 147, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1168] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 150, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1169] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 154, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 155.

[1170] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 158, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 159.

[1171] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 68, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 163.

[1172] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 166, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1173] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 119, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1174] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 168, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1175] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 171, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[1176] In a preferred embodiment, the modified TCR comprises an α chain variable region having an amino acid sequence as shown in SEQ ID NO: 173, and a β chain variable region having an amino acid sequence as shown in SEQ ID NO: 143.

[1177] In a preferred embodiment, the modified TCR comprises an α chain variable region having the amino acid sequence shown in SEQ ID NO: 154, and a β chain variable region having the amino acid sequence shown in SEQ ID NO: 175.

[1178] In some embodiments, the modified TCR further comprises a TCR constant region.

[1179] As used herein, the term "constant region" can be a human constant region or derived from another species, thereby creating a "chimeric" TCR. For example, the human α and / or β chains can be replaced with their murine counterparts ("murinization"), which have been found to enhance surface expression of the human TCR by supporting preferential pairing of the TCR α and β chains and to enhance more stable binding to the CD3 co-receptor.

[1180] In some embodiments, the TCR constant region is a murine constant region or a human constant region.

[1181] It has been reported that the addition of disulfide bonds in the constant region can promote the correct pairing of TCRα and β chains (Kuball J et al. Blood. 2007 Mar 15; 109(6): 2331-8). Therefore, the present invention also contemplates the addition of one or more cysteine ​​modifications in the constant region to form a disulfide bond between the TCRα chain and the TCRβ chain.

[1182] In some embodiments, the TCR constant region includes a TCR α chain constant region and / or a TCR β chain constant region; preferably, the TCR α chain constant region and / or the TCR β chain constant region contain at least one cysteine ​​mutation relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain.

[1183] In some embodiments, the cysteine ​​mutation is at one or more of the following positions: position 48 of the wild-type human TCR α chain constant region, position 48 of the wild-type mouse TCR α chain constant region, position 57 of the wild-type human TCR β chain constant region, and position 57 of the wild-type mouse TCR β chain constant region.

[1184] The sequence of the wild-type TCR constant region can be found in the public database of the International Immunogenetics Information System (IMGT), such as the constant domain sequence of the TCR molecule α chain is "TRAC*01", and the constant domain sequence of the TCR molecule β chain is "TRBC1*01" or "TRBC2*01".

[1185] The positions of the amino acid sequences of the TCR constant regions herein are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT). For example, if an amino acid in the TCR α chain constant region (TRAC) is numbered as position 48 in IMGT, it will be described herein as amino acid position 48 of the TCR α chain constant region (TRAC); if an amino acid in the TCR β chain constant region (TRBC) is numbered as position 57 in IMGT, it will be described herein as amino acid position 57 of the TCR β chain constant region (TRBC); and so on. In the present invention, if the sequence position numbering of other amino acids is specifically indicated, it will be used as indicated.

[1186] In some embodiments, the TCR α chain constant region further comprises an LVL substitution or an LIV substitution, such that the constant region (and / or transmembrane region) comprises the amino acid sequence LLVIVLRIL. For example, when the TCR α chain comprises a human constant region, the human constant region may comprise an LVL mutation such that the constant region (and / or transmembrane region) comprises the amino acid sequence When the TCRα chain comprises a murine constant region, the murine constant region may comprise an LIV mutation such that the constant region (and / or transmembrane region) comprises the amino acid sequence

[1187] In some embodiments, the TCR alpha chain constant region comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:71, and / or the TCR beta chain constant region comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:72.

[1188] In some embodiments, the TCR α chain constant region comprises the amino acid sequence shown in SEQ ID NO:71, and the TCR β chain constant region comprises the amino acid sequence shown in SEQ ID NO:72.

[1189] In some embodiments, the modified TCR may include a signal peptide sequence, for example, a signal peptide sequence may be included at the N-terminus of the α chain and / or the β chain. The signal peptide sequence may be any suitable sequence known in the art. For example, the α chain signal peptide sequence of the TCR of the present invention may have the amino acid sequence (MKTFAGFSFLFLWLQLDCMS) shown in SEQ ID NO: 74. The β chain signal peptide sequence of the TCR of the present invention may have the amino acid sequence (MLLLLLLLGPGSGL) shown in SEQ ID NO: 75.

[1190] In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence as set forth in SEQ ID NO:69, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO:69; and / or a TCR beta chain having an amino acid sequence as set forth in any one of SEQ ID NOs:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176. In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence as shown in SEQ ID NO: 69; and a TCR beta chain having an amino acid sequence as shown in any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176.

[1191] The amino acid sequences of SEQ ID NO: 69 and 3-19 are shown below:

[1192] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, or a sequence identical to or similar to SEQ ID NOs: NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 having a TCR alpha chain with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to any one of SEQ ID NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174; and / or a TCR beta chain with an amino acid sequence as shown in SEQ ID NO:70 or an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to SEQ ID NO:70. In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174; and a TCR beta chain having an amino acid sequence as shown in SEQ ID NO: 70.

[1193] The amino acid sequences of SEQ ID NOs: 20-23, 78 and 70 are shown below:

[1194] In some embodiments, the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, or a sequence identical to or similar to SEQ ID NOs: NO: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 having an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity; and / or having an amino acid sequence selected from any one of SEQ ID NO: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 or a sequence identical to SEQ ID NO: Any of NO:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 has a TCR beta chain with an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity. In some embodiments, the modified TCR comprises a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174; and a TCR beta chain having an amino acid sequence selected from any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176.

[1195] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

[1196] The term "epitope" generally refers to a site on an antigen, typically a (poly)peptide that a binding domain recognizes. The term "binding domain" refers in its broadest sense to an "antigen binding site," i.e., a domain that characterizes a molecule that binds to / interacts with a specific epitope on an antigen target. An antigen target can comprise a single epitope, but typically comprises at least two epitopes, and depending on the size, conformation, and type of the antigen, the antigen target can comprise any number of epitopes. The term "epitope" generally includes linear epitopes and conformational epitopes. A linear epitope is a continuous epitope contained in the primary sequence of amino acids, and it typically includes at least 2 amino acids or more amino acids. A conformational epitope is formed by non-continuous amino acids that are arranged side by side by the folding of the target antigen, and particularly the target (poly)peptide.

[1197] The TCR of the present invention can advantageously (specifically) recognize the MAGE-A1 antigen, in particular the epitope KVLEYVIKV (SEQ ID NO: 2) on MAGE-A1 or a complex of the epitope with an MHC molecule, for example a complex of the epitope with HLA-A*02.

[1198] In some embodiments, the MHC molecule is of the HLA-A*02 type, such as HLA-A*02:01, HLA-A*02:03, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:10, or HLA-A*02: 11. In a preferred embodiment, the MHC molecule is of the HLA-A*02:01 or HLA-A*02:05 type.

[1199] In some embodiments, the modified TCR is soluble or membrane-bound.

[1200] The TCR of the present invention can be provided in the form of a soluble TCR (sTCR). Soluble TCR can be used as a diagnostic tool and a carrier or "adapter" for specifically targeting therapeutic agents or effector cells to, for example, cancer cells expressing an antigen target recognized by the soluble TCR. A soluble TCR is typically a fragment or construct comprising a TCR α chain and / or β chain or its variable region or CDR, and optionally it is stabilized by a disulfide bond or covalently linked by a suitable linker. Typically, a soluble TCR does not include, for example, a transmembrane region.

[1201] The TCR of the present invention can also be provided in a membrane-bound form. Typically, a membrane-bound TCR includes a transmembrane region to anchor it to the cell membrane.

[1202] In some embodiments, the modified TCR is isolated or purified.

[1203] As used herein, the term "isolated or purified" means that the TCR has been identified, separated, and / or recovered from the components of its production environment, such that the "isolated or purified" TCR is free or substantially free of other contaminant components from its production environment that may interfere with its therapeutic or diagnostic use. Contaminant components may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. Thus, an "isolated or purified" TCR can be prepared by at least one purification step that removes or substantially removes these contaminant components.

[1204] In another aspect, the disclosure provides nucleic acids encoding the modified TCRs of the invention.

[1205] In yet another aspect, the disclosure provides a vector comprising a nucleic acid of the invention.

[1206] As used herein, the term "vector" is a nucleic acid molecule used as a medium for transferring (exogenous) genetic material to a host cell, in which the nucleic acid molecule as a vector can be, for example, replicated and / or expressed. The term "vector" encompasses, but is not limited to, plasmids, viral vectors (including retroviral vectors, slow viral vectors, adenoviral vectors, vaccinia virus vectors, polyoma virus vectors, and adenovirus-associated vectors (AAV)), bacteriophages, phagemids, cosmids, and artificial chromosomes (including BACs and YACs). The vector itself is typically a nucleotide sequence, typically a DNA sequence comprising an insert (transgene) and a larger sequence as a vector "skeleton." Engineered vectors are typically included in the starting point (if stable expression of polynucleotides is desired) for autonomous replication in the host cell, a selective marker, and a restriction enzyme cleavage site (such as a multiple cloning site, MCS). The vector can additionally include a promoter, a genetic marker, a reporter gene, a targeting sequence, and / or a protein purification tag. As known to those skilled in the art, a large amount of suitable vectors are known to those skilled in the art, and many are commercially available. Examples of suitable vectors are provided in J. Sambrook et al., Molecular Cloning: A Laboratory Manual (4th ed.), Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, New York (2012), which is incorporated herein by reference in its entirety.

[1207] In some embodiments, the vector is selected from the group consisting of a lentiviral vector, a retroviral vector, a plasmid, a DNA vector, an mRNA vector, a transposon-based vector, and an artificial chromosome.

[1208] In another aspect, the disclosure provides cells comprising a modified TCR, nucleic acid, or vector of the invention.

[1209] As used herein, the term "cell" refers to any type of cell capable of expressing the modified TCR of the present invention. The cell can be a eukaryotic cell, for example, a plant (not having the potential to develop into a plant), an animal, a fungus or an algae, or can be a prokaryotic cell, for example, a bacterium or a protozoan. The cell can be a cultured cell or a primary cell, i.e., directly isolated from an organism, such as a human. The cell can be an adherent cell or a suspension cell, i.e., a cell grown in suspension. Suitable host cells are known in the art and include, for example, DH5α Escherichia coli cells, Chinese hamster ovary cells, monkey VERO cells, COS cells, HEK293 cells, etc. For the purpose of producing the TCR of the present invention, the cell is preferably a mammalian cell. Most preferably, the host cell is a human cell.

[1210] In some embodiments, the cell is selected from the group consisting of a lymphocyte (eg, T cell, NK cell), a monocyte (eg, PBMC), and a stem cell.

[1211] As used herein, the term "stem cell" refers to a stem cell for expressing the TCR of the present invention. For example, stem cells can be lymphoid progenitor cells, induced pluripotent stem cells (iPSCs) or hematopoietic stem cells (HSCs). In some embodiments, stem cells do not include embryonic stem cells obtained by destroying human embryos, and / or do not include totipotent stem cells for developing and forming individual animals. Gene transfer to stem cells generally does not result in TCR expression on the cell surface because CD3 molecules are not expressed on the stem cell surface. However, when stem cells differentiate into lymphoid precursors that migrate to the thymus, the expression of CD3 molecules will start expressing the introduced TCR molecules on the surface of thymocytes.

[1212] In some embodiments, the stem cell is a lymphoid progenitor cell or an induced pluripotent stem cell (iPSC).

[1213] In some embodiments, the cell is a T cell. The T cell can be any T cell, such as a cultured T cell, such as a primary T cell or a T cell from a cultured T cell line, such as Jurkat, SupT1, etc., or a T cell obtained from a mammal. If obtained from a mammal, the T cell can be obtained from many sources, including but not limited to blood, bone marrow, lymph nodes, thymus, or other tissues or liquids. The T cell can also be enriched or purified. Preferably, the T cell is a human T cell. More preferably, the T cell is a T cell separated from people. The T cell can be any type of T cell and can be a T cell of any developmental stage, including but not limited to CD4+ / CD8+ double positive T cells, CD4+ helper T cells, such as Th1 and Th2 cells, CD4+T cells, CD8+T cells (e.g., cytotoxic T cells), tumor infiltrating lymphocytes (TIL), memory T cells (e.g., central memory T cells and effector memory T cells), naive T cells, etc. In some embodiments, the T cell does not express endogenous TCR.

[1214] It is envisioned that the effector cells expressing the TCR of the present invention bind to their antigenic target (preferably an antigenic epitope presented by an antigen presenting cell on HLA-A*02) with high affinity. The term "avidity" or "binding affinity" refers to the ability of a T cell expressing the TCR of the present invention to respond to a given concentration of ligand in vitro, and is believed to be related to the in vivo effector capacity of the cell expressing the TCR. By definition, cells expressing TCRs with high binding affinity respond to very low antigen doses in in vitro assays, while such cells with lower binding affinity require higher amounts of antigen before reaching an immune response similar to that of cells expressing TCRs with high affinity. Therefore, binding affinity can be considered as a quantitative determinant of the activation threshold of cells expressing TCRs. This is determined by exposing such cells to different amounts of cognate antigen in vitro. Cells with high binding affinity TCRs respond to low antigen doses. For example, if a TCR expressing cell activates a target cell when co-cultured with a target antigen peptide-HLA-*02 expressing target cell, the EC50 is no higher than 10 -5 M(such as 10 -5 M or lower, such as 10 -6 M, 10 -7 M, 10 -8 M or 10 -9 M), then the TCR-expressing cell is generally considered to bind its antigen target with "high" binding affinity.

[1215] In yet another aspect, the present disclosure provides a method for preparing the cell of the present invention, comprising the step of transducing or transfecting the cell with the vector of the present invention.

[1216] As used herein, the term "transfection" is a process in which a nucleic acid molecule or polynucleotide (including a vector) is intentionally introduced into a target cell. An example is RNA transfection, which is the process of introducing RNA (e.g., in vitro transcribed RNA, ivtRNA) into a host cell. This term is primarily used for non-viral methods in eukaryotic cells. The term "transduction" is typically used to describe viral-mediated transfer of nucleic acid molecules or polynucleotides. Transfection of animal cells typically involves opening a transient pore or "hole" in the cell membrane to allow the uptake of material. Transfection can be performed using calcium phosphate, by electroporation, by cell extrusion, or by mixing cationic lipids with materials to produce liposomes that fuse with the cell membrane and deposit their cargo inside. Exemplary techniques for transfecting eukaryotic host cells include lipid vesicle-mediated uptake, heat shock-mediated uptake, calcium phosphate-mediated transfection (calcium phosphate / DNA co-precipitation), microinjection, and electroporation.

[1217] In some embodiments, the method further comprises the step of expanding and / or activating the cells before or after the transduction or transfection.

[1218] In another aspect, the present disclosure provides a conjugate comprising a modified TCR according to the present invention and an active agent coupled or conjugated to the TCR.

[1219] In some embodiments, the active agent is selected from a detectable label, an immunostimulatory molecule, and a therapeutic agent. Preferably, the detectable label is selected from biotin, streptavidin, an enzyme or a catalytically active fragment thereof, a radionuclide, a nanoparticle, a paramagnetic metal ion, a nucleic acid probe, a contrast agent, and a fluorescent, phosphorescent, or chemiluminescent molecule. Preferably, the immunostimulatory molecule is selected from a cytokine (such as IL-2 and IFN-γ), a chemokine (such as IL-8), a platelet factor (such as platelet factor 4), and a complement initiator. Preferably, the therapeutic agent is selected from an immunomodulator, a radioactive compound, an enzyme, a chemotherapeutic agent, and a toxin. Other suitable therapeutic agents include small molecule cytotoxic agents, i.e., compounds with the ability to kill mammalian cells with a molecular weight of less than 700 daltons. Such compounds may also contain toxic metals that can have cytotoxic effects. In addition, it should be understood that these small molecule cytotoxic agents also include prodrugs, i.e., compounds that decay or are converted to release cytotoxic agents under physiological conditions. Examples of such agents include cisplatin, maytansine derivatives, rachelmycin, calicheamicin, docetaxel, etoposide, gemcitabine, ifosfamide, irinotecan, melphalan, mitoxantrone, pyroximate, temozolomide, topotecan, trimetrexate glucuronide, auristatin E, vincristine, and doxorubicin; peptide cytotoxins, i.e., proteins or fragments thereof that have the ability to kill mammalian cells; for example, ricin, diphtheria toxin, Pseudomonas bacterial exotoxin A, DNA e and RNase; radionuclides, i.e., unstable isotopes of elements that emit one or more alpha or beta particles or gamma rays while decaying, for example, iodine-131, rhenium-186, indium-111, yttrium-90, bismuth-210 and -213, actinium-225 and astatine-213; chelating agents that can be used to promote the binding of these radionuclides to molecules or polymers thereof; or heterologous protein domains, allologous protein domains, viral / bacterial protein domains, viral / bacterial peptides.

[1220] In yet another aspect, the present disclosure provides bispecific and multispecific antigen-binding molecules comprising a modified TCR or antigen-binding fragment thereof of the present invention, and one or more additional antigen-binding regions that bind one or more additional antigens, wherein the one or more additional antigen-binding regions are each independently selected from an antigen-binding region derived from an antibody and an antigen-binding region derived from a TCR.

[1221] In some embodiments, the one or more additional antigens are each independently selected from a tumor-associated antigen (TAA), a tumor-specific antigen (TSA), a viral antigen, a self-antigen, and an immune cell surface molecule.

[1222] Examples of tumor-associated antigens (TAAs) include, but are not limited to, melanoma-associated antigens (e.g., gp100, MAGEA1, MAGEA3, MAGEA6, MAGEA4, MAGEA2, MAGEA12, MAGEA2B, MAGEA9B, MAGEA10, MAGEA11, MAGEB2, MAGEC1, MAGEC2), IGF2BP1, GNGT1, PI4K2B, CCR8, NPSR1, COX7B2, ONECUT3, SMC1B, FOXI3, GAGE2A, FBXO43, BRDT, PAGE2, GAGE13, POU5F1B, CTAG1A, endogenous reverse transcriptase antigen, 5T4, alpha-fetoprotein, BCMA, CA-125, carcinoembryonic antigen, CSPG4, ROR1, Tyrp-1, TACI, ALK, CLL-1, FLT3, mesothelin, and MUC-1.

[1223] Examples of tumor-specific antigens (TSAs) include, but are not limited to, KRAS, TP53, PIK3CA, CTNNB1, EGFR, BRAF, and GNAS.

[1224] Examples of viral antigens include, but are not limited to, HPV E6 or E7 antigens, CMV antigens, HBV antigens, EBV antigens, herpes virus antigens, human immunodeficiency virus (HIV) antigens, influenza virus antigens, and coronavirus antigens.

[1225] As used herein, the term "autoantigen" refers to an antigenic substance that changes the structure and composition of one's own tissue cells due to biological, physical, or chemical factors, thereby causing the body's immune system to produce an immune response against these self-tissue cell components. The release of hidden antigens can produce an immune response. Certain tissue components in the human body, such as sperm, eye contents, and the brain, never come into contact with immune cells under normal circumstances. When the isolation barrier is destroyed by surgery, trauma, infection, etc., they come into contact with the immune system, thereby producing an immune response, such as autoimmune sympathetic ophthalmia. Changes in one's own components may produce autoantigens. When one's own components are exposed to physical factors (such as cold, heat, ionizing radiation), chemical factors (such as drugs), or biological factors (such as bacteria, viruses, etc.), their antigenicity changes. The altered self-components stimulate the immune system to cause an autoimmune response. For example, denatured IgG can stimulate the body to produce anti-denatured IgG antibodies (rheumatoid factor), causing rheumatoid arthritis. Clinically, the use of certain drugs can change the surface antigenicity of blood cells, causing autoimmune hemolytic anemia or neutropenia. Cross-reactions triggered by shared antigens can generate immune responses. Certain bacteria and viruses share similar antigenic determinants with certain normal human tissue cells. Autoantibodies and sensitized lymphocytes produced against these bacterial and viral antigenic determinants can cross-react with the body's own tissue cells, causing autoimmune diseases. For example, multiple antigenic proteins of group A hemolytic Streptococcus share common antigens with the human myocardium and glomerular basement membrane. The immune response triggered by coxsackievirus infection can attack pancreatic beta cells, causing diabetes.

[1226] In some embodiments, the autoantigen is lupus erythematosus, diabetes caused by autoimmune destruction of pancreatic islets, ankylosing spondylitis, or rheumatoid arthritis.

[1227] Examples of immune cell surface molecules include, but are not limited to, CD3 (e.g., CD3γ, CD3δ, and CD3ε chains), CD4, CD8, CD10, CD11b, CD11c, CD14, CD16, CD18, CD25, CD32a, CD32b, CD41, CD41b, CD42a, CD42b, CD44, CD45RA, CD49, CD61, CD64, CD68, CD94, CD90, CD 117, Nkp46, NKG2D, FcεRI, TCRα / β, TCRγ / δ, HLA-DR, CD28, 4-1BB (CD137), OX40 (CD134), ICOS (CD278), 2B4 (CD244), HVEM, LAG3, DAP10, DAP12, CD27, CD40, GITR, LFA-1, MyD88, CD2, CD7, LIGHT and B7-H3. In some embodiments, the one or more additional antigens are CD3. In some embodiments, the one or more additional antigen binding regions are derived from the antigen binding region of an anti-CD3 antibody or an anti-CD3TCR, preferably from the antigen binding region of an anti-CD3 antibody. In a preferred embodiment, the present disclosure provides a bispecific molecule comprising a modified TCR or antigen binding fragment thereof of the present invention and an antigen binding region of an anti-CD3 antibody.

[1228] In another aspect, the present disclosure provides a composition comprising a modified TCR, nucleic acid, vector, cell, conjugate, or bispecific antigen-binding molecule or multispecific antigen-binding molecule according to the present invention; preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient.

[1229] The term "composition" particularly refers to a composition suitable for administration to humans. However, the term also generally encompasses compositions suitable for administration to non-human animals. The composition and its components (i.e., the active agent and optional carrier or excipient) are preferably pharmaceutically acceptable, i.e., capable of eliciting the desired therapeutic effect in the recipient without causing any undesirable local or systemic effects. The pharmaceutically acceptable compositions of the present invention may, for example, be sterile. Specifically, the term "pharmaceutically acceptable" may mean approved by a regulatory agency or other recognized pharmacopoeia for use in animals, more particularly in humans.

[1230] The term "excipient" includes fillers, binders, disintegrants, coating agents, adsorbents, antiadherents, glidants, preservatives, antioxidants, flavorings, coloring agents, sweeteners, solvents, cosolvents, buffers, chelating agents, viscosity imparting agents, surfactants, diluents, wetting agents, carriers, diluents, preservatives, emulsifiers, stabilizers and tension regulators. It is known to those skilled in the art that suitable excipients are selected to prepare compositions of the present invention. Exemplary carriers for compositions of the present invention include saline, buffered saline, glucose and water. Usually, the selection of suitable excipients especially depends on the desired dosage form of the activating agent used, the disease to be treated and the compositions.

[1231] Depending on the active agent employed (e.g., soluble TCR), the compositions of the present disclosure may be prepared in various forms, such as solid, liquid, gaseous, or lyophilized forms, particularly in the form of ointments, creams, transdermal patches, gels, powders, tablets, solutions, aerosols, granules, pills, suspensions, emulsions, capsules, syrups, liquids, elixirs, extracts, tinctures, or fluid extracts, or in a form particularly suitable for the desired method of administration. Processes known in the art for producing pharmaceuticals are shown in the 22nd edition of Remington's Pharmaceutical Sciences (Ed. Maack Publishing Co, Easton, Pa., 2012), and may include, for example, conventional mixing, dissolving, granulating, sugar-coating, grinding, emulsifying, encapsulating, embedding, or lyophilizing processes. Compositions comprising, for example, host cells or soluble TCRs as described herein are typically provided in liquid form and preferably contain a pharmaceutically acceptable buffer.

[1232] In some embodiments, the composition further comprises a second therapeutic agent, preferably, the second therapeutic agent is selected from an antibody, a chemotherapeutic agent, and a small molecule drug.

[1233] Preferred examples of the second therapeutic agent include known anticancer drugs such as cisplatin, maytansine derivatives, rachelmycin, calicheamicin, docetaxel, etoposide, gemcitabine, ifosfamide, irinotecan, melphalan, mitoxantrone, sorfimer sodium photofrin II, temozolomide, topotecan, trimetreate glucuronate, auristatin E, E), vincristine and doxorubicin; and peptide cytotoxins, such as ricin, diphtheria toxin, Pseudomonas bacterial exotoxin A, DNA enzymes and RNA enzymes; radionuclides, such as iodine-131, rhenium-186, indium-111, iridium-90, bismuth-210 and 213, actinium-225 and astatine-213; prodrugs, such as antibody-directed enzyme prodrugs; immunostimulants, such as IL-2, chemokines such as IL-8, platelet factor 4; antibodies or fragments thereof, such as anti-CD3 antibodies or fragments thereof; complement activators; viral / bacterial protein domains and viral / bacterial peptides.

[1234] In yet another aspect, the present disclosure provides a method of treating a MAGE-A1 positive cancer in a subject, comprising administering to the subject an effective amount of a modified TCR of the present invention.

[1235] In another aspect, the present disclosure provides a method of treating a MAGE-A1 positive cancer in a subject, comprising administering to the subject an effective amount of the cells of the present invention.

[1236] As used herein, the term "treatment" includes therapeutic or prophylactic treatment in a subject in need thereof. "Therapeutic or prophylactic treatment" includes prophylactic treatment intended to completely prevent clinical and / or pathological manifestations or therapeutic treatment intended to ameliorate or alleviate clinical and / or pathological manifestations. Thus, the term "treatment" also includes ameliorating or preventing a disease.

[1237] As used herein, the term "effective amount" means an amount of a therapeutic agent that, when administered to a subject for the treatment or prevention of a disease, is sufficient to achieve such treatment or prevention. The "effective amount" may vary depending on the compound, the disease and its severity, and the age, weight, etc. of the subject to be treated. A "therapeutically effective amount" refers to an effective amount for therapeutic treatment. A "prophylactically effective amount" refers to an effective amount for preventive treatment.

[1238] Therapeutic efficacy and toxicity can be determined by standard procedures such as ED50 (the dose therapeutically effective in 50% of the population) and LD50 (the dose lethal to 50% of the population) in cell cultures or experimental animals. The dose ratio between therapeutic and toxic effects is the therapeutic index and can be expressed as the ratio of ED50 / LD50. Pharmaceutical compositions that exhibit a large therapeutic index are preferred.

[1239] Those skilled in the art can determine the exact dosage of the TCR or cell administered using known techniques. A suitable dosage provides a sufficient amount of the active agent of the present invention, and is preferably therapeutically effective, i.e., sufficient to cause, for example, a therapeutic or preventive response in a subject or animal within a reasonable timeframe. For example, the dosage of the TCR of the present invention should be sufficient to bind to cancer antigens or detect, treat or prevent cancer within a period of about 2 hours or longer, such as 12 hours to 24 hours or longer (e.g., 1 month, 2 months, 3 months, 6 months, 12 months, 24 months, etc.) from the time of administration. In certain embodiments, the time period can be even longer. As known in the art, for therapeutic purposes (such as alleviating acute attacks relative to the disease), route of administration, time and frequency, time and frequency of administration of the formulation, age, weight, general health, sex, diet, severity of the disease state, drug combination, reaction sensitivity and adjustment of tolerance / response to treatment may be necessary.

[1240] Many assays for determining the dosage to be administered are known in the art. For the purposes of the present invention, assays can be used to determine the starting dose to be administered to a mammal, comprising administering a given dose of T cells expressing the TCR of the present invention to a group of mammals (each administered a different dose of T cells), comparing the extent of target cell lysis or IFN-γ secreted by such T cells. After administering a certain dose, the extent of target cell lysis or IFN-γ secretion achieved can be determined by methods known in the art. The dosage of the TCR or cells of the present invention can also be determined by the presence, nature, and extent of any adverse side effects that may accompany the administration of the TCR or cells of the present invention. Typically, the attending physician decides the dosage of the TCR or cells of the present invention administered to each individual patient, taking into account a variety of factors such as age, weight, general health, diet, sex, active agent to be administered, route of administration, and severity of the treatment condition. In some embodiments of the methods of treatment disclosed herein, the number of cells administered per infusion can, for example, range from about 1×10 6 to about 1×10 12 In some embodiments, less than 1x10 6 cells.

[1241] It will be appreciated that treatment may require a single administration of a therapeutically effective dose or multiple administrations of a therapeutically effective dose of an active agent of the invention. For example, depending on the formulation, half-life, and clearance rate of the particular composition, some compositions may be administered every 3 to 4 days, weekly, or once every two weeks, or once in a month.

[1242] The compositions of the present disclosure can be applied in a variety of ways. Typically, administration is accomplished parenterally. Parenteral delivery methods include topical, intra-arterial, intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, intrauterine, intravaginal, sublingual, or intranasal administration.

[1243] The terms "subject," "individual," "animal," or "patient" are used interchangeably herein to refer to any subject in need of treatment, particularly a mammalian subject. Generally, mammalian subjects include humans, non-human primates, dogs, cats, guinea pigs, rabbits, rats, mice, horses, cows, dairy cows, and the like. However, it will be readily appreciated that the TCRs, nucleic acids, vectors, host cells, and pharmaceutical compositions provided herein are specifically contemplated for use in treating human subjects, particularly those that are HLA-A*02 positive, e.g., HLA-A*02:01, HLA-A*02:03, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:10, or HLA-A*02:11 positive, preferably HLA-A*02:01 or HLA-A*02:05 positive.

[1244] In some embodiments of the disclosed treatment methods, the cells are autologous or allogeneic to the subject.

[1245] In some embodiments, the method comprises the following steps: (i) isolating a sample containing cells from the subject; (ii) transducing or transfecting the cells with a vector of the present invention; and (iii) administering the cells obtained in step (ii) to the subject. In some embodiments, the method further comprises the step of knocking out the endogenous TCR in the cells after step (i) and before (ii).

[1246] In some embodiments, the MAGE-A1 positive cancer is selected from melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial bladder carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[1247] In some embodiments, the method further comprises administering a second therapeutic agent, preferably, the second therapeutic agent is selected from an antibody, a chemotherapeutic agent, and a small molecule drug. Preferred examples of the second therapeutic agent are as described above.

[1248] In some embodiments of the disclosed treatment methods, the subject has an HLA-A*02 allele, such as an HLA-A*02:01, HLA-A*02:03, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:10, or HLA-A*02:11 allele, preferably an HLA-A*02:01 or HLA-A*02:05 allele.

[1249] In another aspect, the present disclosure provides a method for detecting (e.g., diagnosing) a MAGE-A1 positive cancer in a subject, wherein the method comprises (i) contacting a sample obtained from the subject with a TCR, cell, or conjugate of the invention; and (ii) detecting the presence of a MAGE-A1 antigen in the sample, wherein the presence of the MAGE-A1 antigen indicates the MAGE-A1 positive cancer. The MAGE-A1 positive cancer is preferably selected from melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial carcinoma of the bladder), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[1250] In some embodiments, the sample obtained from the subject can be a blood sample, a urine sample, a tissue sample, or a cell sample. In certain embodiments, the method is performed in vitro. In some embodiments, the method comprises (i) contacting the sample obtained from the subject with a conjugate of the present disclosure, wherein the conjugate comprises a detectable label; and (ii) detecting the presence of the MAGE-A1 antigen in the sample by detecting the detectable label. Examples of detectable labels include, but are not limited to, biotin, streptavidin, an enzyme or a catalytically active fragment thereof, a radionuclide, a nanoparticle, a paramagnetic metal ion, a nucleic acid probe, a contrast agent, and a fluorescent, phosphorescent, or chemiluminescent molecule; preferably an enzyme or a catalytically active fragment thereof, a radionuclide, a fluorescent, phosphorescent, or chemiluminescent molecule.

[1251] In another aspect, the present disclosure provides a kit comprising the TCR or conjugate of the present invention, wherein the kit is used to detect the presence of a positive epitope in a test sample, wherein the epitope is an epitope comprising KVLEYVIKV (SEQ ID NO: 2).

[1252] In some embodiments, the kit is a kit for detecting (e.g., diagnosing) a MAGE-A1 positive cancer in a subject, comprising a TCR or conjugate of the present invention. MAGE-A1 positive cancers are preferably selected from melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial bladder carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[1253] In some embodiments, the conjugate comprises a detectable label. Examples of detectable labels include, but are not limited to, biotin, streptavidin, an enzyme or catalytically active fragment thereof, a radionuclide, a nanoparticle, a paramagnetic metal ion, a nucleic acid probe, a contrast agent, and a fluorescent, phosphorescent, or chemiluminescent molecule; preferably, an enzyme or catalytically active fragment thereof, a radionuclide, a fluorescent, phosphorescent, or chemiluminescent molecule. In some embodiments, the kit may further comprise instructions for use of the kit.

[1254] In another aspect, the present disclosure provides use of a modified TCR, nucleic acid, vector, cell, conjugate, bispecific antigen-binding molecule or multispecific antigen-binding molecule, or composition of the invention in the preparation of a medicament for treating a MAGE-A1-positive cancer in a subject.

[1255] In yet another aspect, the disclosure provides a modified TCR, nucleic acid, vector, cell, conjugate, bispecific antigen-binding molecule, or multispecific antigen-binding molecule, or composition of the invention for use in treating a MAGE-A1 positive cancer in a subject.

[1256] In another aspect, the present disclosure provides use of the modified TCR or conjugate of the present invention in preparing a kit for detecting the presence of a positive epitope in a test sample, wherein the epitope is an epitope comprising KVLEYVIKV (SEQ ID NO: 2).

[1257] In some embodiments, the present disclosure provides use of a modified TCR or conjugate of the present invention in the preparation of a kit for detecting (eg, diagnosing) a MAGE-A1-positive cancer in a subject.

[1258] In another aspect, the present disclosure provides a modified TCR or conjugate of the present invention for detecting the presence of a positive epitope in a test sample, wherein the epitope is an epitope comprising KVLEYVIKV (SEQ ID NO: 2), for example, detecting (e.g., diagnosing) MAGE-A1 positive cancer in a subject.

[1259] In some embodiments of the uses disclosed herein, the MAGE-A1-positive cancer is preferably selected from melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial bladder carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[1260] In some embodiments, the subject has an HLA-A*02:01 allele, such as an HLA-A*02:01, HLA-A*02:03, HLA-A*02:05, HLA-A*02:06, HLA-A*02:07, HLA-A*02:10, or HLA-A*02:11 allele, preferably an HLA-A*02:01 or HLA-A*02:05 allele.

[1261] The present invention will be further described below in conjunction with specific examples, and the advantages and features of the present invention will become clearer as the description progresses. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, were performed according to conventional conditions in the art, for example, the conditions described in Sambrook and Russeii et al., Molecular Cloning: A Laboratory Manual (3rd Edition) (2001), CSHL Press, or according to the conditions recommended by the manufacturer. Unless otherwise stated, the experimental materials and reagents used in the following examples are commercially available.

[1262] Example 1. Screening for affinity-enhanced substituted TCRs

[1263] The present inventors used the human TCR ht27 sequence as a template, substituted the complementarity determining region (CDR) region of ht27 by in vitro substitution, and obtained high-affinity substitution clones by in vitro positive screening of the substituted TCR. Through this method, the preferred TCR sequence screened was at the positions Gln29, Ala36, Thr37, Thr38 of CDR1β of the ht27β chain, Ser56, Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65 of CDR2β, and Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114 of CDR3β. and Glu115 contain amino acid substitutions, specifically amino acid substitutions include Q29P, A36I, T37E, T38R, S56T, N57D, N57E, N57H, N57K, N57Q, N57R, N57S, N57T, E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, E58W, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, G59T, G59P, G59L, S63D, S63E, S63H, S63K, S63Q, S63R, S63T, K64M, K64C, K64W, K64T, K64P, K64A, A65F, A65I, A65L, A65M, A65P, A65V, A65W, A65Y, S105L, S105N, S105H, S105Q, S105K, S105R, E108D, E108F, E10 8T, E108I, E108K, E108P, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, Q111H, G112E, P113H, EPGQGP at positions 108-113 are replaced by VNDGT, Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, Y114S, E115I, etc.The selected preferred TCR sequences contain amino acid substitutions at positions Asp27, Ser28, Ser29, Ser36, Thr37, and Tyr38 of CDR1α and positions Ile56, Phe57, Ser58, Asn59, Met63, and Asp64 of CDR2α of ht27α chain. The specific amino acid substitutions include S28Y, S28P, S29Y, S29T, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29Y. 9N, S36M, S36L, S36Q, S36I, S36F, S36P, S36V, S36D, T37I, Y38L, I56T, I56G, F57W, F57T, S58G, S58I, S58N, S58A, S58P, S58W, N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, N59T, M63D, M63Q, M63T, M63E, D64T, D64Y, D64S, etc. The preferred TCR sequences screened also include amino acid substitutions at the Thr26 position of the α chain framework region 1 (FR1α), such as T26A, T26V, etc.

[1264] The three CDR regions of each TCR α and β chain bind to different sites on the MHC-peptide complex, interacting with the complex in a relatively independent manner. Mutations in these CDR regions, whether alone or in combination, effectively increase TCR affinity relative to wild-type TCRs. Some combinations of mutations can even produce additive effects, achieving even higher affinity.

[1265] Representative substituted TCRs are shown in Table 1. Table 1 shows the amino acid substitution regions and substitution types of representative substituted TCRs of the present invention relative to wild-type ht27 TCR.

[1266] Table 1. Representative substituted TCRs

[1267] The inventors further combined the mutations in the above-mentioned different CDR regions and screened out mutant TCRs with affinity up to the pM level. The amino acid substitution positions of the screened TCR sequences include positions Asp27, Phe28, Gln29, Ala36, Thr37, Thr38 of CDR1β of ht27β chain, positions Ser56, Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65 of CDR2β, and positions Ser105, Ala106, Arg107, Glu108, Pro109, Gl Amino acid substitutions at positions Y110, Gln111, Gly112, Pro113, Tyr114, and Glu115, as well as positions Asp27, Ser28, Ser29, Ser36, Thr37, and Tyr38 in the CDR1α of the ht27α chain, and positions Ile56, Phe57, Ser58, Asn59, Met63, Asp64, and Met65 in the CDR2α, also including amino acid substitutions at position Thr26 in FR1α. Representative substituted TCRs are shown in Table 2 below. Table 2 shows the amino acid substitution regions and substitution types of representative substituted TCRs of the present invention relative to the wild-type ht27 TCR.

[1268] Table 2. Representative replacement TCRs

[1269] The CDR sequences and amino acid sequences of the α and β chain variable regions of the wild-type ht27 TCR are shown in Table 3. The CDR sequences and amino acid sequences of the α and β chain variable regions of representative substituted TCRs of the present invention are shown in Table 4.

[1270] Table 3. CDR sequences and amino acid sequences of the α and β chain variable regions of ht27 TCR

[1271] Table 4. Representative substituted TCR CDR sequences and amino acid sequences of the α and β chain variable regions

[1272] To increase the correct pairing and expression of exogenous TCR on the surface of human T cells, the variable regions of the α and β chains of TCR were fused to the murine TCR constant region, and the classic α chain C region "LIV" substitution was introduced into the murine TCR constant region (i.e., amino acid substitutions were introduced into the murine TCR α chain constant region so that the constant region contained the amino acid sequence in bold) and the canonical α chain T48C and β chain S57C substitutions (in bold and underlined).

[1273] Amino acid sequence of the α chain constant region:

[1274] Amino acid sequence of the β chain constant region:

[1275] The α and β chains of the TCR (the N-termini of the α and β chains contain the signal peptide sequences MKTFAGFSFLFLWLQLDCMS (SEQ ID NO: 74) and MLLLLLLLGPGSGL (SEQ ID NO: 75), respectively) were constructed in series into a lentiviral expression vector using the 2A self-cleavage peptide (ATNFSLLKQAGDVEENPGP (SEQ ID NO: 73)) in the order of β-T2A-α. Taking the ht27-WT TCR as an example, the amino acid sequence of the tandem TCR molecule is shown below (the signal peptide is shown in bold, the constant region sequence is underlined, and the 2A self-cleavage peptide is shown in italics):

[1276] The natural human TCR ht27WT and the replacement TCR sequence of the present invention were cloned into TCR tandem expression lentiviral vectors, packaged with lentivirus, transduced into reporter cells JKR9, and then sorted for TCR expression-positive cells. HLA-A*02:01-positive T2 cells were loaded with different concentrations of MAGE-A1 target antigen peptide (KVLEYVIKV (SEQ ID NO: 2)), and then co-incubated with transduced TCR ht27WT and the JKR9 reporter cells replacing TCR of the present invention for activation. After 16 hours of activation, the reporter gene was stained for flow cytometry analysis. The activation results of the high-affinity TCR of the present invention on reporter cells are shown in Figures 1 and 2. According to the results of Figure 2, the EC50 value was calculated, the EC50 value of ht27-WT was set to 1, and the EC50 enhancement multiple of the replacement TCR relative to ht27-WT was calculated, and the results are shown in Table 5.

[1277] Table 5. EC50 values ​​of representative substituted TCRs and their EC50 fold enhancement relative to wild-type ht27 TCR

[1278] Among them, the activation function of the mutant TCR obtained by combining mutations in different CDR regions is significantly better than that of the wild-type TCR ht27, with an affinity as high as the pM level, and its EC50 value is shown in Table 6 below.

[1279] Table 6. EC50 values ​​of combined mutant TCRs

[1280] The above results show that TCRs with amino acid substitutions at one or more of amino acid positions Asn57, Glu58, Gly59, Ser63, Lys64, Ala65, Ser105, Gln111, Tyr114, Ser29, Ser36, and Asn59 exhibit significantly improved affinity relative to wild-type TCR ht27. These amino acid substitutions can be further combined to generate substituted TCRs with higher affinity.

[1281] However, after transduction of JKR9 reporter cells with the combined mutant TCRs, nonspecific activation reactions occurred with T2 cells not loaded with target peptides (Figure 1), which may be due to the excessively high affinity of these TCRs. Similar nonspecific activation was also observed using flow cytometry analysis as described above using ultra-high affinity mutant TCRs targeting gp100 known in the art (gp100-b03a8, gp100-b28a2, gp100-b28a8, gp100-b33a2, gp100-b17a2, gp100-b10a2, gp100-b23a2, gp100-b26a2, gp100-b26a8, gp100-b27a2, gp100-b27a8) (Figure 3). Despite nonspecific activation, the soluble TCR protein drug Kimmtrak (trade name: Tebentafusp-tebn), prepared using the mutant TCR gp100-b27a8, has been approved for marketing for the treatment of metastatic uveal melanoma. This demonstrates the medicinal value of ultra-high affinity mutant TCRs (KD-SPR value <1nM).

[1282] Example 2. Assessment of the affinity of ultra-high affinity mutant TCRs by Dextramer staining

[1283] Dextramers are polymers of pMHC molecules. In in vitro experiments, they can be used to identify TCR sequences that specifically recognize specific antigenic peptides from T cell libraries. The specific binding of dextramers to TCRs can be detected and sorted using flow cytometry through fluorescent labeling. Furthermore, the strength of dextramer binding to TCRs is positively correlated with TCR affinity. Therefore, dextramer staining can be used to screen for ultra-high-affinity mutant TCRs and assess their affinity.

[1284] To validate the feasibility of this approach, experiments were performed using mutant TCRs with known affinities for the target gp100 (gp100-b03a8, gp100-b28a2, gp100-b28a8, gp100-b33a2, gp100-b17a2, gp100-b10a2, gp100-b23a2, gp100-b26a2, gp100-b26a8, gp100-b27a2, and gp100-b27a8). The affinities of the known gp100-TCR mutants, as determined by SPR, were as follows:

[1285] The verification method is as follows:

[1286] First, a gp100 mutant TCR with known affinity was transduced into JKR9 cells via lentivirus to construct a mutant TCR-expressing cell line. Dextramer was then prepared using the gp100 target antigen peptide (YLEPGPVTV, SEQ ID NO: 177); the Dextramer was then diluted 10-fold with PBS, with a total of four concentration gradients (the final staining concentration was 2nM to 0.002nM). Subsequently, JKR9 cells transduced with mutant TCRs of different affinities were stained in parallel with different concentrations of Dextramer; the staining positivity rate and MFI of each experimental group were recorded by flow cytometry. The results are shown in Figure 4. Finally, by summarizing the data from different experimental groups, the Dextramer staining positivity rate and mean staining intensity (MFI) were analyzed to see if there was a correlation between the affinity measured by SPR.

[1287] The results showed that as the concentration of Dextramer used during staining decreased, the advantage of high-affinity TCR staining gradually emerged, and both the staining positivity rate and MFI showed a staining advantage related to the affinity of the mutant TCR (Figure 4). After extracting the staining data, further analysis found that when the concentration of Dextramer was lower than 0.2nM, the staining intensity showed a linear correspondence with the affinity of the TCR (Figure 5). The EC50 of the extreme low concentration (0.02nM) Dextramer staining MFI and the Dextramer gradient concentration staining MFI were correlated with the true affinity (KD-SPR) of the SPR assay, respectively. It was found that the extreme low concentration Dextramer staining MFI and the Dextramer gradient concentration staining MFI could well reflect the SPR assay results, and the linear relationship formula between KD-SPR and Dextramer staining MFI was obtained (Figure 6). These results show that Dextramer staining can be used to evaluate the affinity of mutant TCRs.

[1288] After verification of the above method, the affinity of the mutant TCR targeting MAGE-A1 of the present invention was evaluated using the above Dextramer staining method. The results are shown in Figures 7 and 8.

[1289] The results showed that the mutant TCR of the present invention showed strong positive staining under extremely low concentration Dextramer staining conditions, and the staining intensity (MFI) increased with increasing Dextramer concentration, indicating that the mutant TCR of the present invention has extremely high affinity for the target-pMHC complex.

[1290] Using the linear relationship formula for KD-SPR and Dextramer staining MFI described above ( FIG. 6 ), the Dextramer staining MFI of the mutant TCRs of the present invention was substituted into the formula to estimate the KD-SPR values ​​of the mutant TCRs. The results are shown in the table below. The results show that the estimated SPR values ​​for all mutant TCRs reached the 10 pM level, demonstrating the extremely high affinity of the mutant TCRs of the present invention.

[1291] Example 3. Evaluation of cell membrane expression stability of affinity-optimized TCR

[1292] Lentiviral vectors (MOI = 10) encoding ht27WT and affinity-optimized TCRs were packaged in 293T cells. PBMCs were transduced and expanded until harvest on day 11. At harvest, cells showed good proliferation, with a proliferation fold ranging from 70-150 times, and cell viability >90%. T cells were stained with human CD3-FITC antibody to confirm successful TCR transduction. TCR-T expression was assessed with anti-mouse TCRβ-APC antibody. CD4 / CD8 ratios were assessed with CD4-APC and CD8-PE-Cy7 antibodies.

[1293] Flow cytometry analysis showed that under identical transduction and culture conditions, the CD4 / CD8 ratio of each cell line after affinity-optimized TCR transduction was close to 1:1, and the TCR positivity rate was greater than 75%, ranging from 75-90% (Figure 9). This demonstrates that affinity-optimized TCR molecules possess excellent membrane expression stability, suggesting their potential for strong antigen reactivity and anti-tumor activity.

[1294] Example 4. Efficient and specific killing of antigen-positive tumor cells by affinity-optimized TCR-T cells

[1295] T cells expressing affinity-optimized TCRs or ht27WT were used as effector cells, and PBMCs that were not transduced with TCRs were expanded and cultured in parallel as negative controls for effector cells. Hela-A2-eGFP-LUC (MAGE-A1 positive, HLA-A*02:01 positive) were used as HLA-antigen peptide-matched positive target cells; Siha-eGFP-LUC cells (MAGE-A1 negative, HLA-A*02:01 negative) were used as HLA-antigen peptide-mismatched negative target cells. All target cells stably expressed the luciferase gene. Effector cells were incubated with different target cells at effector-target ratios (E:T) of 9:1, 3:1, and 1:1 for 16-20 hours, and luciferase substrate was added to detect surviving target cells. The proportion of target cells killed was calculated based on the remaining target cells, and the results are shown in Figure 10.

[1296] The results showed that affinity-optimized TCR-T cells had significant cytotoxicity against Hela-A2-eGFP-LUC-positive target cells, and their cytotoxicity was significantly higher than that of Ht27WT TCR-T cells. In contrast, affinity-optimized TCR-T cells had no cytotoxicity against negative target cells. These results indicate that affinity-optimized TCR-T cells have the ability to specifically and efficiently kill HLA-antigen peptide-matched target cells, and their cytotoxicity increases with increasing affinity.

[1297] Example 5. Specific INF-γ secretion by affinity-optimized TCR-T cells against antigen-positive tumor cells

[1298] T cells expressing affinity-optimized TCRs or ht27WT were used as effector cells, and PBMCs that were not transduced with TCRs were expanded and cultured in parallel as negative controls for effector cells. Hela-A2-eGFP-LUC and A375-eGFP-LUC cells (MAGE-A1 positive, HLA-A*02:01 positive) were used as positive target cells for HLA-antigen peptide matching; Siha-eGFP-LUC cells (MAGE-A1 negative, HLA-A*02:01 negative) were used as negative target cells for HLA-antigen peptide mismatch. Effector cells were incubated with different target cells at an effector-target ratio (E:T) of 1:1 for 16-20 hours, and IFN-γ secretion in the supernatant was detected using an IFN-γ ELISA kit (ExCell, Cat#EH008-96). The results are shown in Figure 11.

[1299] The results showed that affinity-optimized TCR-T cells could be detected to secrete significant IFN-γ after co-incubation with positive target cells A375 or Hela-A2, and the amount of IFN-γ secreted was significantly higher than that of Ht27WT TCR-T cells. However, no IFN-γ secretion was detected for negative target cells with mismatched HLA-antigen peptides. These results indicate that affinity-optimized TCR-T cells have good specific killing effect on HLA-antigen peptide-matched target cells, and this specific killing effect is enhanced with the increase of affinity.

[1300] Example 6. Nonspecific activation of T2-KO-TAP1 cell lines expressing different HLA types by affinity-optimized TCR-T cells

[1301] To test whether affinity-optimized TCR-T cells can produce alloreactivity, that is, antigen-independent cross-reactions with other mismatched HLAs, a T2-KO-TAP1 cell line (HLA knockout and expression of antigen peptide transporter TAP1) was constructed. This cell line expresses 66 different HLAs, and the selected high-frequency HLA-A, HLA-B, and HLA-C can cover more than 90% of the population. After transduction of different HLA genes into T2-KO-TAP1 cells, the HLA expression rate was higher than 95% after flow cytometry sorting. 1x10 5 T cells expressing affinity-optimized TCRs were used as effector cells and 1x10 5 Target cells (T2-KO-TAP1 cells expressing different HLA) were co-cultured for 16-24 hours, and the supernatant was collected for IFN-γ detection.-7 T2 cells (HLA-A*02:01 positive) expressing the MAGE-A1 polypeptide of M were used as a positive control of target cells. The results are shown in FIG12 .

[1302] The results showed that no IFN-γ secretion was detected after co-culture of Ht27M1001 (affinity increased 388-fold) and Ht27M1005 (affinity increased 11-fold) with all T2-KO-TAP1-HLA cells, indicating that Ht27M1001 and Ht27M1005 have good specificity.

[1303] Example 7. Nonspecific activation of affinity-optimized TCRs against antigen-negative target cells from different tissues and T2-KO-TAP1 cell lines expressing different HLA typing

[1304] To further confirm the specificity of affinity-optimized TCRs in recognizing target antigens, we tested the nonspecific activation of affinity-optimized TCRs against antigen-negative target cells from different tissues and T2-KO-TAP1 cell lines expressing different HLA types. The specific experimental procedures are as follows.

[1305] Construction of a tool for off-target reaction detection: Reference was made to the TPM (Transcripts Per Million mapped reads) of transcriptome sequencing data from the Cancer Cell Line Encyclopedia (CCLE) database, with a target gene TPM of 0 defined as negative target gene expression. Nine cell lines derived from different tissues that were negative for MAGE-A1 gene expression were selected as antigen-negative target cells. These cell lines were transduced with the HLA-A*02:01 gene and used as tool cells for off-target reaction detection.

[1306] Constructing an alloreactivity detection tool: Knockout the endogenous HLA-B*51:01 and HLA-C*01:02 genes in T2 cells, rendering them unable to express endogenous class I HLA molecules (T2KO cells); then overexpress the TAP1 / 2 genes in T2 cells to construct tool cells with complete endogenous antigen presentation function (T2KO-TAP); finally, transduce different high-frequency HLAs into T2KO-TAP cells to construct T2-KO-TAP1 cell lines of different typing as tool cells for detecting alloreactivity. The specific construction was performed as described in Example 6.

[1307] Non-specific reactivity detection: During the detection process, the JKR9 reporter cells transducing the affinity-optimized TCR were co-cultured with antigen-negative target cells overexpressing HLA-A*02:01 and T2-KO-TAP1 cell lines expressing different HLA typing. After 16 hours of culture, flow cytometry was used to determine whether the JKR9 reporter cells transduced with the high-affinity mutant TCR could be activated by the above tool cells, and then to determine whether the mutant TCR had off-target reactions or allogeneic reactions. HLA-antigen peptide-matched positive target cells A375-A1-A2 cells (MAGE-A1 positive, HLA-A*02:01 positive) were used as positive controls. The non-specific activation results of the high-affinity TCR of the present invention are shown in the following table.

[1308] The results showed that ht27M1001, ht27M1002, ht27M1003, ht27M1004, ht27M1005, ht27M1006, ht27M1009, ht27M1010, ht27M1011, ht27M1012, ht27M1013, ht27M1018, ht27M1019, ht27M1128, ht27M1166, ht27M1169, ht271170, ht27M1226, ht27M1240, ht27M1243, ht27M1245, ht27M1285, and ht27M1290 , ht27M1369, ht27M1378, ht27M1414, ht27M1457, ht27M1557, ht27M1387, ht27M1398, ht27M1401, ht27M1374, ht27M1381, ht27M1360, ht27M1362, ht27M1363, ht27M1365, ht27M1418, and ht27M1436 did not detect non-specific activation of HLA-matched antigen-negative target cells and T2-KO-TAP1 cell lines expressing different HLA typing, indicating that these affinity-optimized TCRs have good specificity.

[1309] This application makes reference to various issued patents, published patent applications, journal articles and other publications, all of which are incorporated herein by reference. If any incorporated reference conflicts with this specification, this specification shall prevail. In addition, any specific embodiment of the present invention that falls within the scope of the prior art may be explicitly excluded from any one or more claims. Because the embodiments are considered to be known to those skilled in the art, they may be excluded, even if the exclusion is not explicitly listed in this application. Any specific embodiment of the present invention may be excluded from any claim for any reason, regardless of whether it is related to the existence of the prior art.

[1310] Although the present invention has been described with reference to specific embodiments thereof, it should be understood by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adapt a particular situation, material, composition, method, or process step to the objective, spirit, and scope of the present invention. All such modifications are intended to be within the scope of the appended claims.

Claims

1. A modified T cell receptor (TCR), comprising an α chain variable region comprising CDR1α, CDR2α, and CDR3α, and a β chain variable region comprising CDR1β, CDR2β, and CDR3β, wherein the modified TCR comprises the following amino acid substitutions relative to a parent TCR: (i) amino acid substitutions in the variable region of the β chain comprising (a) amino acid substitution at one or more of the following positions in CDR1β: Gln29, Ala36, Thr37 and Thr38, (b) amino acid substitutions at one or more of the following positions in CDR2β: Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65, and / or (c) amino acid substitutions at one or more of the following positions in CDR3β: Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115; and / or (ii) amino acid substitutions in the α chain variable region comprising (d) amino acid substitution at one or more of the following positions in CDR1α: Asp27, Ser28, Ser29, Ser36, Thr37 and Tyr38, (e) amino acid substitutions in CDR2α at the following positions: Ile56, Phe57, Ser58, Asn59, Met63, and Asp64, and / or (f) amino acid substitution at Thr26 in FR1α; wherein the parent TCR comprises a CDR1β with an amino acid sequence as set forth in SEQ ID NO: 24, a CDR2β with an amino acid sequence as set forth in SEQ ID NO: 25, a CDR3β with an amino acid sequence as set forth in SEQ ID NO: 26, a CDR1α with an amino acid sequence as set forth in SEQ ID NO: 27, a CDR2α with an amino acid sequence as set forth in SEQ ID NO: 28, and a CDR3α with an amino acid sequence as set forth in SEQ ID NO: 29; The amino acid positions are numbered according to the nomenclature of the International Immunogenetics Information System (IMGT).

2. The modified TCR according to claim 1, wherein the modified TCR comprises one or more of the following amino acid substitutions (1)-(33) relative to the parent TCR: (1) an amino acid substitution at position Gln29 in CDR1β selected from the group consisting of Q29E and Q29P; (2) amino acid substitution A36I at position Ala36 in CDR1β; (3) amino acid substitution T37E at position Thr37 in CDR1β; (4) amino acid substitution T38R at position Thr38 in CDR1β; (5) amino acid substitution S56T at position Ser56 in CDR2β; (6) an amino acid substitution at position Asn57 in CDR2β selected from the group consisting of N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T; (7) an amino acid substitution at position Glu58 in CDR2β selected from the group consisting of E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W; (8) an amino acid substitution at position Gly59 in CDR2β selected from the group consisting of G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T; (9) an amino acid substitution at position Ser63 in CDR2β selected from the group consisting of S63D, S63E, S63H, S63K, S63Q, S63R, and S63T; (10) an amino acid substitution at position Lys64 in CDR2β selected from the group consisting of K64Q, K64M, K64C, K64W, K64T, K64P, and K64A; (11) Amino acid substitution at position Ala65 in CDR2β, selected from A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y; (12) an amino acid substitution at position Ser105 in CDR3β selected from the group consisting of S105L, S105N, S105H, S105Q, S105K, and S105R; (13) an amino acid substitution at position Glu108 in CDR3β selected from the group consisting of E108D, E108F, E108T, E108I, E108K, and E108P; (14) amino acid substitution G110W at position Gly110 in CDR3β; (15) an amino acid substitution at position Gln111 in CDR3β selected from the group consisting of Q111D, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H; (16) amino acid substitution of EPGQGP to VNDGT at positions Glu108 to Pro113 in CDR3β; (17) an amino acid substitution at position Gly112 in CDR3β selected from the group consisting of G112P, G112A, and G112E; (18) an amino acid substitution at position Pro113 in CDR3β selected from the group consisting of P113W and P113H; (19) an amino acid substitution at position Tyr114 in CDR3β selected from the group consisting of Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S; (20) an amino acid substitution at position Glu115 in CDR3β selected from the group consisting of E115D and E115I; (21) an amino acid substitution at position Asp27 in CDR1α selected from the group consisting of D27C and D27Y; (22) an amino acid substitution at position Ser28 in CDR1α selected from the group consisting of S28M, S28A, S28G, S28Y, and S28P; (23) an amino acid substitution at position Ser29 in CDR1α selected from the group consisting of S29E, S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N; (24) an amino acid substitution at position Ser36 in CDR1α selected from the group consisting of S36D, S36M, S36L, S36Q, S36I, S36F, S36P, and S36V; (25) amino acid substitution T37I at position Thr37 in CDR1α; (26) amino acid substitution Y38L at position Tyr38 in CDR1α; (27) an amino acid substitution at position Ile56 in CDR2α selected from the group consisting of I56T and I56G; (28) an amino acid substitution at position Phe57 in CDR2α selected from the group consisting of F57W and F57T; (29) an amino acid substitution at position Ser58 in CDR2α selected from the group consisting of S58G, S58I, S58N, S58A, S58P, and S58W; (30) an amino acid substitution at position Asn59 in CDR2α selected from the group consisting of N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T; (31) an amino acid substitution at position Met63 in CDR2α selected from the group consisting of M63D, M63Q, M63T, and M63E; (32) an amino acid substitution at position Asp64 in CDR2α selected from the group consisting of D64A, D64Q, D64T, D64Y, and D64S; (33) An amino acid substitution at the Thr26 position in FR1α selected from the group consisting of T26S, T26I, T26P, T26Q, T26A, and T26V.

3. The modified TCR according to claim 1 or 2, wherein the modified TCR comprises one or more of the following amino acid substitutions (1)-(4) in CDR1β relative to the parent TCR: (1) Q29E or Q29P; (2)A36I; (3)T37E; (4)T38R; Preferably, the modified TCR comprises the amino acid substitution Q29E or Q29PA36IT37ET38R in CDR1β relative to the parent TCR.

4. The modified TCR according to any one of claims 1 to 3, wherein the modified TCR comprises one or more of the following amino acid substitutions (1) to (7) in CDR2β relative to the parent TCR: (1)S56T; (2) N57Q or N57D; (3) E58L, E58P, or E58Y; (4) G59Q, G59P, or G59L; (5) S63T or S63D; (6) K64C or K64Q; (7)A65V; Preferably, the modified TCR comprises the amino acid substitution N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L, S56TN57DE58Y or S63DK64Q in CDR2β relative to the parent TCR.

5. The modified TCR according to any one of claims 1 to 4, wherein the modified TCR comprises one or more of the following amino acid substitutions (1) to (8) in CDR3β relative to the parent TCR: (1)G110W; (2) Q111D, Q111G, or Q111S; (3) G112E, G112P or G112A; (4) P113H or P113W; (5) Y114S, Y114F, Y114L or Y114W; (6) E115I or E115D; (7) S105L, S105Q, S105N or S105H; (8) EPGQGP at positions Glu108 to Pro113 was substituted with VNDGT; Preferably, the modified TCR comprises the following amino acid substitutions in CDR3β relative to the parent TCR: G110WQ111DG112PP113W, G112AP113WY114SE115D, Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, EPGQGP at positions Glu108 to Pro113 replaced by VNDGT or G112EP113HY114LE115I.

6. The modified TCR according to any one of claims 1 to 5, wherein the modified TCR comprises one or more of the following amino acid substitutions (1) to (3) relative to the parent TCR: (1) amino acid substitutions Q29E or Q29PA36IT37ET38R in CDR1β; (2) amino acid substitutions in CDR2β: N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L, S56TN57DE58Y, or S63DK64Q; (3) Amino acid substitutions in CDR3β: G110WQ111DG112PP113W, G112AP113WY114SE115D, Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, and EPGQGP at positions Glu108 to Pro113 were substituted with VNDGT or G112EP113HY114LE115I.

7. The modified TCR according to any one of claims 1 to 6, wherein the modified TCR comprises one or more of the following amino acid substitutions (1) to (7) in CDR1α and / or FR1α relative to the parent TCR: (1) D27C or D27Y; (2) S28Y, S28P, S28M, S28A or S28G; (3) S29T, S29L, S29Y, S29F, S29E or S29M; (4) S36F, S36D, S36M, S36L or S36Q; (5)T37I; (6)Y38L; (7) T26A, T26V, T26S, T26I, T26P or T26Q; Preferably, the modified TCR comprises the following amino acid substitutions in CDR1α and / or FR1α relative to the parent TCR: S29TS36DT37IY38L, T26AS28YS29L, T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F or T26QD27YS28GS29M.

8. The modified TCR according to any one of claims 1 to 7, wherein the modified TCR comprises one or more of the following amino acid substitutions (1) to (6) in CDR2α relative to the parent TCR: (1)I56T or I56G; (2) F57W or F57T; (3) S58G, S58I, S58N, S58A, S58P or S58W; (4) N59F, N59T, N59H or N59L; (5) M63D, M63Q, M63T or M63E; (6) D64A, D64Q, D64T, D64Y or D64S; Preferably, the modified TCR comprises the amino acid substitutions N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F in CDR2α relative to the parent TCR.

9. The modified TCR according to any one of claims 1 to 8, wherein the modified TCR comprises one or more of the following amino acid substitutions (1) to (2) relative to the parent TCR: (1) CDR1α and / or FR1α contain the following amino acid substitutions: S29TS36DT37IY38L, T26AS28YS29L, T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F, or T26QD27YS28GS29M; (2) CDR2α contains the amino acid substitutions N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L, or I56TS58WN59F.

10. The modified TCR according to claim 1, wherein the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

11. The modified TCR according to claim 1, wherein the modified TCR comprises: (i) a β chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively; and (ii) an α chain variable region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively.

12. The modified TCR according to claim 1, wherein the modified TCR comprises: (i) an α chain variable region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 109 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 112 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 115 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 121 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 124 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 127 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 140, 141 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 146, 118 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 149 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 165, 118 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 118 and 29, respectively, or CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 141 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 170, 76 and 29, respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 118 and 29, respectively, or CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 152, 149 and 29, respectively; and (ii) a β chain variable region comprising CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 36, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 30 and 37, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 38, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 39, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 40, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 41, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 79, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 82, 25 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 85 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 88 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25 and 94, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 25 and 131, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 91 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 153 and 35, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 26, respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 130, 91 and 162, respectively, or CDR1β, CDR2β and CDR3β have the amino acid sequences shown in SEQ ID NOs: 82, 91 and 33, respectively.

13. The modified TCR according to claim 1 , wherein the modified TCR comprises an α chain variable region having an amino acid sequence as set forth in SEQ ID NO: 46, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 46, and a β chain variable region having an amino acid sequence as set forth in any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175.

14. The modified TCR according to claim 1, wherein the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 or a sequence identical to SEQ ID NOs: NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 having an α chain variable region with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to any one of SEQ ID NO:65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, and a β chain variable region having an amino acid sequence as set forth in SEQ ID NO:47, or an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to SEQ ID NO:

47.

15. The modified TCR according to claim 1, wherein the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 65-68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 or a sequence identical to SEQ ID NOs: NO:98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173 having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity, and an α chain variable region having an amino acid sequence selected from any one of SEQ ID NO:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 or a sequence identical to SEQ ID NO: Any of NOs:48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175 has a beta chain variable region with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

16. The modified TCR according to any one of claims 1 to 15, wherein the modified TCR further comprises a TCR constant region.

17. The modified TCR according to claim 16, wherein the TCR constant region is a murine constant region or a human constant region.

18. The modified TCR according to claim 16 or 17, wherein the TCR constant region comprises a TCR α chain constant region and / or a TCR β chain constant region; Preferably, the TCR α chain constant region and / or the TCR β chain constant region comprises at least one cysteine ​​substitution relative to the wild-type sequence to form a disulfide bond between the TCR α chain and the TCR β chain; Preferably, the TCR α chain constant region comprises an LVL substitution or an LIV substitution, such that the α chain constant region comprises the amino acid sequence LLVIVLRIL.

19. The modified TCR according to claim 16, wherein the modified TCR comprises a TCR alpha chain having an amino acid sequence as shown in SEQ ID NO: 69, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to SEQ ID NO: 69, and a TCR beta chain having an amino acid sequence as shown in any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176, or an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176.

20. The modified TCR according to claim 16, wherein the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, or a sequence identical to or similar to SEQ ID NOs: NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 having a TCR alpha chain with an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to any one of SEQ ID NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, and a TCR beta chain with an amino acid sequence shown in SEQ ID NO:70 or an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 95% or at least 99% identical to SEQ ID NO:

70.

21. The modified TCR according to claim 16, wherein the modified TCR comprises an amino acid sequence selected from any one of SEQ ID NOs: 20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, or a sequence identical to or similar to SEQ ID NOs: NO:20-23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 having an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity, and a TCR alpha chain having an amino acid sequence selected from any one of SEQ ID NO:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 or a sequence identical to SEQ ID NO: Any of NO:3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 has a TCR beta chain with an amino acid sequence of at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity.

22. The modified TCR according to any one of claims 1 to 21, wherein the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of the epitope with an MHC molecule.

23. The modified TCR according to claim 22, wherein the MHC molecule is of HLA-A*02 type, such as HLA-A*02:01 type or HLA-A*02:05 type.

24. The modified TCR according to any one of claims 1 to 23, wherein the modified TCR is soluble or membrane-bound.

25. The modified TCR according to any one of claims 1-24, wherein the modified TCR is isolated or purified.

26. A nucleic acid encoding a modified TCR according to any one of claims 1 to 25.

27. A vector comprising the nucleic acid according to claim 26.

28. The vector according to claim 27, wherein the vector is selected from the group consisting of a lentiviral vector, a retroviral vector, a plasmid, a DNA vector, an mRNA vector, a transposon-based vector and an artificial chromosome.

29. A cell comprising a modified TCR according to any one of claims 1 to 25, a nucleic acid according to claim 26, or a vector according to claim 27 or 28.

30. The cell according to claim 29, wherein the cell is selected from the group consisting of lymphocytes (eg, T cells, NK cells), monocytes (eg, PBMCs), and stem cells.

31. The cell according to claim 30, wherein the stem cell is a lymphoid progenitor cell or an induced pluripotent stem cell (iPSC).

32. The cell according to claim 30, wherein the cell is a T cell.

33. The cell according to claim 32, wherein the T cell does not express endogenous TCR.

34. A method for preparing the cell according to any one of claims 29 to 33, comprising the step of transducing or transfecting the cell with the vector according to claim 27 or 28.

35. The method according to claim 34, further comprising the step of expanding and / or activating the cells before or after said transduction or transfection.

36. A conjugate comprising a modified TCR according to any one of claims 1 to 25 and an active agent coupled or conjugated to the TCR.

37. The conjugate according to claim 36, wherein the active agent is selected from the group consisting of a detectable label, an immunostimulatory molecule, and a therapeutic agent; Preferably, the detectable label is selected from the group consisting of biotin, streptavidin, an enzyme or catalytically active fragment thereof, a radionuclide, a nanoparticle, a paramagnetic metal ion, a nucleic acid probe, a contrast agent, and a fluorescent, phosphorescent or chemiluminescent molecule; Preferably, the immunostimulatory molecules are selected from cytokines, chemokines, platelet factors and complement initiators; Preferably, the therapeutic agent is selected from the group consisting of an immunomodulator, a radioactive compound, an enzyme, a chemotherapeutic agent, and a toxin.

38. A bispecific or multispecific antigen binding molecule comprising a modified TCR or antigen binding fragment thereof according to any one of claims 1 to 25, and one or more additional antigen binding regions that bind one or more additional antigens, optionally wherein the one or more additional antigen binding regions are each independently selected from an antigen binding region derived from an antibody and an antigen binding region derived from a TCR.

39. The bispecific or multispecific antigen binding molecule according to claim 38, wherein the one or more additional antigens are each independently selected from a tumor-associated antigen (TAA), a tumor-specific antigen (TSA), a viral antigen, an autoantigen, and an immune cell surface molecule.

40. The bispecific or multispecific antigen binding molecule according to claim 39, wherein the one or more additional antigens is CD3, preferably the one or more additional antigen binding regions are derived from an antigen binding region of an anti-CD3 antibody.

41. A composition comprising a modified TCR according to any one of claims 1 to 25, a nucleic acid according to claim 26, a vector according to claim 27 or 28, a cell according to any one of claims 29 to 33, a conjugate according to claim 36 or 37, or a bispecific or multispecific antigen-binding molecule according to any one of claims 38 to 40; Preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient.

42. The composition according to claim 41, wherein the composition further comprises a second therapeutic agent, preferably, the second therapeutic agent is selected from the group consisting of an antibody, a chemotherapeutic agent and a small molecule drug.

43. A method of treating a MAGE-A1 positive cancer in a subject comprising administering to the subject an effective amount of a modified TCR according to any one of claims 1-25.

44. A method of treating a MAGE-A1 positive cancer in a subject comprising administering to the subject an effective amount of a cell according to any one of claims 29-33.

45. The method according to claim 44, wherein the cells are autologous or allogeneic to the subject.

46. ​​The method according to claim 44, comprising the steps of: (i) isolating a sample containing cells from the subject; (ii) transducing or transfecting the cell with the vector of claim 27 or 28; and (iii) administering the cells obtained in step (ii) to a subject.

47. The method according to claim 46, further comprising the step of knocking out the endogenous TCR in the cell after step (i) and before step (ii).

48. The method according to any one of claims 43 to 47, wherein the MAGE-A1 positive cancer is selected from the group consisting of melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., urothelial bladder carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, bile duct cancer, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

49. The method according to any one of claims 43 to 48, wherein the method further comprises administering a second therapeutic agent, preferably, the second therapeutic agent is selected from the group consisting of an antibody, a chemotherapeutic agent and a small molecule drug.

50. The method according to any one of claims 43 to 49, wherein the subject has an HLA-A*02 allele, such as an HLA-A*02:01 or HLA-A*02:05 allele.

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