MAGE-A1-specific TCR and its uses

Abstract

The present disclosure provides an MAGE-A1-specific TCR and its use, in particular, a TCR modified to improve binding affinity, a nucleic acid encoding the same, a vector containing the nucleic acid of the present invention, a cell containing the TCR, nucleic acid or vector of the present invention, and a method for producing the cell of the present invention. The present disclosure further relates to a conjugate or composition containing the TCR of the present invention, and a method for treating cancer by using the TCR or cell of the present invention.

Description

Technical Field

[0001] This international patent application claims the benefit of priority of Chinese Patent Application CN202210622198.1, filed on June 1, 2022, the entire content of which is incorporated herein by reference.

[0002] The present disclosure relates to T cell receptors (TCRs) and their use, particularly TCRs against MAGE-A1, and their use in the treatment of MAGE-A1 positive cancers.

Background Art

[0003] TCR-T involves cloning a TCR that specifically recognizes a specific HLA-tumor antigen peptide complex by corresponding experimental techniques, and then delivering the TCR gene coding sequence to more T cells by gene delivery means such as lentivirus, thereby conferring new antigen recognition specificity to these T cells. Patient-derived T cells can efficiently recognize tumor cell-specific antigens by transducing the TCR gene in vitro and proliferating it in large quantities. These T cells can be returned to the patient to specifically kill tumor cells and exert anti-tumor activity. Since either intracellular antigens or extracellular antigens can be recognized by TCRs via HLA presentation, TCR-T can target most tumor-specific antigens. In particular, since it can recognize those intracellular tumor antigens (about 90% of the total antigens), TCR-T is promising for use in the treatment of almost all tumors, particularly various solid tumors.

[0004] Numerous studies have found that tumor cells express certain tumor-associated antigens on their surface, and these tumor-associated antigens are not expressed or are expressed at low levels in normal tissue cells. Therefore, developing TCR-T products targeting tumor-associated antigens is an important means of tumor immunotherapy. Some of the tumor-associated antigens are only directed towards expression on the surface of tumor cells and germ cells known as "cancer-testis antigens", and such antigen genes are, of course, preferred targets for TCR-T product research. The melanoma-associated antigen gene family MAGE has been the most studied cancer-testis antigen so far, and MAGE-A1 is the first identified cancer-testis antigen gene. Numerous studies have shown that MAGE-A1 is expressed on the surface of various solid tumor cells such as melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, etc. Therefore, the development of MAGE-A1 TCR-T products can benefit more people with various indications.

[0005] HLA-A0201-restricted TCRs targeting MAGE-A1 have previously been developed under the names "ht27" and "T1367" respectively (Obenaus, M., Leitao, C., Leisegang, M., Chen, X., Gavvovidis, I., van der Bruggen, P., Uckert, W. et al., Identification of human T‐cell receptors with optimal affinity to cancer antigens using antigen‐negative humanized mice. Nat. Biotechnol. 2015. 33: 402-407; Ottaviani, S., Zhang, Y., Boon, T. and van der 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. ht27 is a natural TCR sequence found in humans, but this TCR has low affinity and it is difficult to effectively exert its anti-tumor effect in vivo. “T1367” is a mouse-derived TCR sequence found in transgenic mice, targeting the human MAGE-A1 antigen. This TCR has high affinity to efficiently kill tumor cells in in vitro experiments, but its application to human experiments may cause a strong autoimmune reaction with no therapeutic effect. Therefore, it is necessary to develop a TCR product that specifically targets MAGE-A1 with high affinity.

Summary of the Invention

Problems to be Solved by the Invention

[0006] Using the human-derived TCR ht27 sequence as a template, the complementarity-determining regions (CDRs) of ht27 were mutated using in vitro mutagenesis, and the mutated TCRs were screened positively in vitro to obtain mutant clones with high affinity. Then, negative screening was performed in vitro to exclude the presence of non-specifically activated mutant clones, thereby obtaining a mutant TCR sequence with high affinity and no non-specific reaction. Also, through in vitro experiments, the specificity and non-specific reaction of the mutant TCR sequence were verified, and finally, a mutant TCR sequence with strong specificity and high affinity was obtained.

[0007] Thus, in one form, the present disclosure provides a modified T cell receptor (TCR) comprising a variable α-chain region comprising CDR1α, CDR2α, and CDR3α and a variable β-chain region comprising CDR1β, CDR2β, and CDR3β, having the following amino acid substitutions compared to the parental TCR: (i) An amino acid substitution in the variable region of the β-chain, which (a) includes an amino acid substitution at one or more positions of Asn57, Glu58, Gly59, Ser63, Lys64, Ala65 in CDR2β, and / or (b) includes an amino acid substitution at one or more positions of Ser105, Glu108, Gln111 and Tyr114 in CDR3β, and / or (ii) An amino acid substitution in the variable region of the α-chain, which (c) includes an amino acid substitution at one or more positions of Ser29 and Ser36 in CDR1α, (d) includes an amino acid substitution at the position of Asn59 in CDR2α,

[0008] wherein the parent TCR comprises a CDR1β with the amino acid sequence shown in SEQ ID NO:24, a CDR2β with the amino acid sequence shown in SEQ ID NO:25, a CDR3β with the amino acid sequence shown in SEQ ID NO:26, a CDR1α with the amino acid sequence shown in SEQ ID NO:27, a CDR2α with the amino acid sequence shown in SEQ ID NO:28, and a CDR3α with the amino acid sequence shown in SEQ ID NO:29, and provides a modified TCR numbered according to the nomenclature rules of the International ImMunoGeneTics Information System (IMGT).

[0009] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(13) compared to the parent TCR: (1) An amino acid substitution at the position of Asn57 selected from N57D, N57E, N57H, N57K, N57Q, N57R, N57S and N57T; (2) An amino acid substitution at the position of Glu58 selected from E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y and E58W; (3) An amino acid substitution at the position of Gly59 selected from G59D, G59E, G59H, G59K, G59Q, G59R, G59S and G59T; (4) An amino acid substitution at position Ser63 selected from S63D, S63E, S63H, S63K, S63Q, S63R, and S63T; (5) Selected from K64M, K64C, K64W, K64T, K64P, and K64A An amino acid substitution at position Lys64; (6) An amino acid substitution at position Ala65 selected from A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y; (7) An amino acid substitution at position Ser105 selected from S105L, S105N, S105H, S105Q, S105K, and S105R; (8) An amino acid substitution at position Glu108 selected from E108D, E108F, E108T, E108I, E108K, and E108P; (9) An amino acid substitution at position Gln111 selected from Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H; (10) An amino acid substitution at position Tyr114 selected from Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S; (11) An amino acid substitution at position Ser29 selected from S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N; (12) An amino acid substitution at position Ser36 selected from S36M, S36L, S36Q, S36I, S36F, S36P, and S36V; (13) An amino acid substitution at position Asn59 selected from N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T; Comprising one or more of the above.

[0010] In some embodiments, the modified TCR does not contain the S105N amino acid substitution alone compared to the parental TCR.

[0011] In some embodiments, the modified TCR comprises, compared to the parental TCR, one or more of the following amino acid substitutions in CDR2β: N57D, E58Y, G59Q, S63T, K64C, and A65V; preferably, the modified TCR comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β compared to the parental TCR.

[0012] In some embodiments, the modified TCR comprises, compared to the parental TCR, the following amino acid substitutions (1) to (3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H; comprising one or more of: preferably, the modified TCR comprises the amino acid substitutions Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, or S105N in CDR3β compared to the parental TCR.

[0013] In some embodiments, the modified TCR comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β and the amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F, or Q111SY114F in CDR3β compared to the parental TCR.

[0014] In some embodiments, the modified TCR comprises, compared to the parental TCR, the following (1) and / or (2) in CDR1α: (1) S29L; (2) S36M, S36L, or S36Q.

[0015] In some embodiments, the modified TCR comprises the amino acid substitution N59F in CDR2α compared to the parental TCR.

[0016] In some embodiments, the modified TCR is (i) an α-chain variable region comprising CDR1α, CDR2α, and CDR3α having the amino acid sequences set forth in SEQ ID NOs: 27-29, respectively, and (ii) a β-chain variable region, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 30, and 26, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 31, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 32, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 33, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 34, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 35, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β having the amino acid sequences set forth in SEQ ID NOs: 24, 30, and 37, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, or The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, and a variable β-chain region containing the same.

[0017] In some embodiments, the modified TCR (i) A variable β-chain region containing the CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24 to 26, respectively, and (ii) A variable α-chain region, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76, and 29, respectively, and a variable α-chain region containing the same. In some embodiments, the modified TCR (i) A variable α-chain region, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28, and 29, respectively, The CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 28 and 29 respectively, The CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28 and 29 respectively, or The CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76 and 29 respectively, comprising a variable α-chain region, and (ii) A variable β-chain region, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30 and 26 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 31 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 32 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 33 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 34 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 35 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30 and 33 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30 and 34 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30 and 35 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 36 respectively, The CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 37 respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, CDR1β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, CDR2β and CDR3β, or CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, and includes a variable β-chain region.

[0018] In some embodiments, the modified TCR includes a variable α-chain region having the amino acid sequence 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 variable β-chain region having the 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.

[0019] In some embodiments, the modified TCR comprises an α-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 65 to 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 to 68 and 77, and a β-chain variable region having an amino acid sequence shown 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.

[0020] In some embodiments, the modified TCR comprises an α-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 48 to 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 to 64, and a β-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 48 to 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 to 64.

[0021] 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 comprises 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.

[0022] In some embodiments, the modified TCR comprises a TCR α-chain having the amino acid sequence 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 β-chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs:3 to 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 to 19.

[0023] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs:20 to 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 to 23 and 78, and a TCR β-chain having the amino acid sequence shown 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.

[0024] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in 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 β-chain having an amino acid sequence selected from the amino acid sequences 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.

[0025] 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 and an MHC molecule.

[0026] In some embodiments, the MHC molecule is HLA-A*02 type, such as HLA-A*02:01 type or HLA-A*02:05 type.

[0027] In some embodiments, the modified TCR is soluble or membrane-bound.

[0028] In some embodiments, the modified TCR is isolated or purified.

[0029] In another form, the present disclosure provides a modified T cell receptor (TCR) comprising a variable α-chain region comprising CDR1α, CDR2α and CDR3α, and a variable β-chain region comprising CDR1β, CDR2β and CDR3β, wherein the modified TCR has the following amino acid substitutions compared to the parental TCR: (i) An amino acid substitution in the variable β-chain region, (a) An amino acid substitution at one or more positions of Gln29, Ala36, Thr37, Thr38 in CDR1β, (b) one or more amino acid substitutions at positions Ser56, Asn57, Glu58, and Gly59 in CDR2β, and / or (c) one or more amino acid substitutions at positions Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115 in CDR3β, and / or (ii) an amino acid substitution in the variable region of the α-chain, (d) one or more amino acid substitutions at positions Ser28, Ser29, Ser36, Thr37, and Tyr38 in CDR1α, (e) one or more amino acid substitutions at positions Ile56, Phe57, Ser58, Asn59, Met63, and Asp64 in CDR2α, and / or (f) an amino acid substitution at position Thr26 in FR1α, comprising wherein the parental TCR has a CDR1β with the amino acid sequence shown in SEQ ID NO:24 , a CDR2β with the amino acid sequence shown in SEQ ID NO:25, a CDR3β with the amino acid sequence shown in SEQ ID NO:26, a CDR1α with the amino acid sequence shown in SEQ ID NO:27, a CDR2α with the amino acid sequence shown in SEQ ID NO:28, and a CDR3α with the amino acid sequence shown in SEQ ID NO:29, wherein the amino acid positions are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT), providing a modified TCR.

[0030] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(25) compared to the parental TCR: (1) an amino acid substitution Q29P at position Gln29 in CDR1β; (2) an amino acid substitution A36I at position Ala36 in CDR1β; (3) an amino acid substitution T37E at position Thr37 in CDR1β; (4) an amino acid substitution T38R at position Thr38 in CDR1β; (5) an amino acid substitution S56T at position Ser56 in CDR2β; (6) An amino acid substitution at the Asn57 position in CDR2β selected from N57Q and N57D; (7) An amino acid substitution at the Glu58 position in CDR2β selected from E58L, E58P, and E58Y; (8) An amino acid substitution at the Gly59 position in CDR2β selected from G59P and G59L; (9) An amino acid substitution of EPGQGP to VNDGT at positions from Glu108 to Pro113 in CDR3β; (10) An amino acid substitution G112E at the Gly112 position in CDR3β; (11) An amino acid substitution P113H at the Pro113 position in CDR3β; (12) An amino acid substitution Y114L at the Tyr114 position in CDR3β; (13) An amino acid substitution E115I at the Glu115 position in CDR3β; (14) An amino acid substitution at the Ser28 position in CDR1β selected from S28Y and S28P; (15) An acid substitution at the Ser29 position in CDR1β selected from S29T, S29L, and S29Y; (16) An amino acid substitution S36D at the Ser36 position in CDR1α; (17) An amino acid substitution T37I at the Thr37 position in CDR1α; (18) An amino acid substitution Y38L at the Tyr38 position in CDR1α; (19) An amino acid substitution at the Ile56 position in CDR2α selected from I56T and I56G; (20) An amino acid substitution at the Phe57 position in CDR2α selected from F57W and F57T; (21) An amino acid substitution at the Ser58 position in CDR2α selected from S58G, S58I, S58N, S58A, S58P, and S58W; (22) An amino acid substitution at the Asn59 position in CDR2α selected from N59F, N59T, N59H, and N59L; (23) An amino acid substitution at the Met63 position in CDR2α selected from M63D, M63Q, M63T, and M63E; (24) An amino acid substitution at the Asp64 position in CDR2α, selected from D64T, D64Y, and D64S; (25) An amino acid substitution at the Thr26 position in FR1α, selected from T26A and T26V; Comprising one or more of the following.

[0031] In some embodiments, the modified TCR, compared to the parental TCR, Comprises one or more of the following amino acid substitutions in CDR1β: Q29P, A36I, T37E, and T38R. Preferably, the modified TCR, compared to the parental TCR, comprises the amino acid substitutions Q29P, A36I, T37E, and T38R in CDR1β.

[0032] In some embodiments, the modified TCR, compared to the parental TCR, Comprises one or more of the following amino acid substitutions (1)-(4) in CDR2β: (1) S56T; (2) N57Q or N57D; (3) E58L, E58P, or E58Y; (4) G59P or G59L; Preferably, the modified TCR, compared to the parental TCR, comprises the amino acid substitutions S56TE58LG59P, S56TN57QE58PG59L, or S56TN57DE58Y in CDR2β.

[0033] In some embodiments, the modified TCR, compared to the parental TCR,

[0034] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1) to (6) in CDR1α and / or FR1α as compared to the parental TCR: (1) S28Y or S28P; (2) S29T, S29L or S29Y; (3) S36D; (4) T37I; (5) Y38L; (6) T26A or T26V; and preferably, the modified TCR comprises amino acid substitutions: S29T S36D T37I Y38L, T26A S28Y S29L or T26V S28P S29Y in CDR1α and / or FR1α as compared to the parental TCR.

[0035] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1) to (6) in CDR2α as compared to the parental 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) D64T, D64Y or D64S; and preferably, the modified TCR comprises amino acid substitutions S58G N59F M63D D64T, S58I N59F M63Q D64Y, S58N N59T M63Q, N59F M63T D64Y, S58A N59F M63E D64S, I56T F57W S58P N59H, I56G F57T S58W N59L or I56T S58W N59F in CDR2α as compared to the parental TCR.

[0036] In some embodiments, the modified TCR ​​(i) An α-chain variable region comprising CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27 to 29, respectively, and (ii) A β-chain variable region, 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, or A β-chain variable region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24, 25, and 94, respectively. In some embodiments, the modified TCR (i) A β-chain variable region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24 to 26, respectively, and (ii) An α-chain variable region, 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, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29 respectively, or An α-chain variable region comprising the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29 respectively.

[0037] In some embodiments, the modified TCR is (i) An α-chain variable region comprising The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 97, 28, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 100, 28, and 29 respectively, The CDR1α of the amino acid sequences shown in SEQ ID NO: 103, 28, and 29 respectively , CDR2α, and CDR3α, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 106, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 109, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, or An α-chain variable region comprising the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively, and (ii) A β-chain variable region, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 79, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 25, and 26, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 85, and 26, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 88, and 26, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 26, respectively, or A β-chain variable region comprising the CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 94, respectively.

[0038] In some embodiments, the modified TCR comprises an α-chain variable region having an amino acid sequence 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 β-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NO: 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 NO: 80, 83, 86, 89, 92, 95.

[0039] In some embodiments, the modified TCR comprises an α-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NO: 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 NO: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and a β-chain variable region having an amino acid sequence shown 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.

[0040] In some embodiments, the modified TCR has an α-chain variable region having an amino acid sequence shown in any one of SEQ ID NO: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, or an amino acid sequence selected from amino acid sequences 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 NO: 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and a β-chain variable region having an amino acid sequence shown in any one of SEQ ID NO: 80, 83, 86, 89, 92, 95, or an amino acid sequence selected from amino acid sequences 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 NO: 80, 83, 86, 89, 92, 95.

[0041] 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 comprises 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.

[0042] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence 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 β-chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NO: 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 NO: 81, 84, 87, 90, 93, 96.

[0043] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NO: 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 NO: 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, and a TCR β-chain having an amino acid sequence shown 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.

[0044] In some embodiments, the modified TCR has an amino acid sequence shown in any one of SEQ ID NO: 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 NO: 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, and a TCRα chain having an amino acid sequence selected from the group consisting of: an amino acid sequence shown in any one of SEQ ID NO: 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 NO: 81, 84, 87, 90, 93, 96 and a TCRβ chain having an amino acid sequence selected from the group consisting of: 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 NO: 81, 84, 87, 90, 93, 96.

[0045] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2), or a complex of said epitope and an MHC molecule.

[0046] In some embodiments, the MHC molecule is of the HLA-A*02 type, such as HLA-A*02:01 or HLA-A*02:05.

[0047] In some embodiments, the modified TCR is soluble or membrane-bound.

[0048] In some embodiments, the modified TCR is isolated or purified.

[0049] In yet another form, the present disclosure provides a modified T cell receptor (TCR) comprising a variable region of the α chain comprising CDR1α, CDR2α and CDR3α, and a variable region of the β chain comprising CDR1β, CDR2β and CDR3β, wherein the modified TC has the following amino acid substitutions as compared to the parental TCR: (i) An amino acid substitution in the variable region of the β-chain, where (a) an amino acid substitution at one or more positions of Gln29, Ala36, Thr37, Thr38 in CDR1β, (b) an amino acid substitution at one or more positions of Ser56, Asn57, Glu58, Ser63 and Lys64 in CDR2β, and / or (c) an amino acid substitution at one or more positions of Gly110, Gln111, Gly112, Pro113, Tyr114 and Glu115 in CDR3β, and / or (ii) an amino acid substitution in the variable region of the α-chain, where (d) an amino acid substitution at one or more positions of Asp27, Ser28, Ser29, Ser36 in CDR1α, (e) an amino acid substitution at the positions of Ser58, Asn59, Met63 and Asp64 in CDR2α, and / or (f) an amino acid substitution at the position of Thr26 in FR1α, including wherein the parental TCR comprises a CDR1β with the amino acid sequence shown in SEQ ID NO:24, a CDR2β with the amino acid sequence shown in SEQ ID NO:25, a CDR3β with the amino acid sequence shown in SEQ ID NO:26, a CDR1α with the amino acid sequence shown in SEQ ID NO:27, a CDR2α with the amino acid sequence shown in SEQ ID NO:28, and a CDR3α with the amino acid sequence shown in SEQ ID NO:29, wherein the amino acid positions are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT), providing a modified TCR.

[0050] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(24) compared to the parental TCR: (1) An amino acid substitution at the position of Gln29 selected from Q29E and Q29P; (2) The amino acid substitution A36I at the position of Ala36; (3) The amino acid substitution T37E at the position of Thr37; (4) The amino acid substitution T38R at the position of Thr38; (5)Amino acid substitution S56T at position Ser56; (6)Amino acid substitution N57D at position Asn57; (7)Amino acid substitution E58Y at position Glu58; (8)Amino acid substitution S63D at position Ser63; (9)Amino acid substitution K64Q at position Lys64; (10)Amino acid substitution G110W at position Gly110; (11)Amino acid substitution Q111D at position Gln111; (12)Amino acid substitution at position Gly112 selected from G112P and G112A; (13)Amino acid substitution P113W at position Pro113; (14)Amino acid substitution at position Tyr114 selected from Y114W, Y114S and Y114F; (15)Amino acid substitution E115D at position Glu115; (16)Amino acid substitution at position Asp27 selected from D27C and D27Y; (17)Amino acid substitution at position Ser28 selected from S28P, S28M, S28A and S28G; (18)Amino acid substitution at position Ser29 selected from S29Y, S29L, S29F, S29E and S29M; (19)Amino acid substitution at position Ser36 selected from S36L, S36Q, S36M and S36F; (20)Amino acid substitution at position Ser58 selected from S58I and S58A; (21)Amino acid substitution N59F at position Asn59; (22)Amino acid substitution at position Met63 selected from M63Q and M63E; (23)Amino acid substitution at position Asp64 selected from D64A, D64S and D64Q; (24)Amino acid substitution at position Thr26 selected from T26V, T26S, T26I, T26P and T26Q; comprises one or more of the following.

[0051] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(4) in CDR1β compared to the parental TCR: (1) Q29E or Q29P; (2) A36I; (3) T37E; (4) T38R; and preferably, the modified TCR comprises the amino acid substitution Q29E or Q29P A36I T37E T38R in CDR1β compared to the parental TCR. Preferably, the modified TCR comprises the amino acid substitution Q29E or Q29P A36I T37E T38R in CDR1β compared to the parental TCR.

[0052] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions in CDR2β compared to the parental TCR: S56T, N57D, E58Y, S63D, and K64Q, and preferably, the modified TCR comprises the amino acid substitution S56T N57D E58Y or S63D K64Q in CDR2β compared to the parental TCR.

[0053] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(6) in CDR3β compared to the parental TCR: (1) G110W; (2) Q111D; (3) G112P or G112A; (4) P113W; (5) Y114W, Y114S, or Y114F; (6) E115D; and preferably, the modified TCR comprises the amino acid substitution: G110W Q111D G112P P113W, Y114W, G112A P113W Y114S E115D, or Y114F in CDR3β compared to the parental TCR. Preferably, the modified TCR comprises the amino acid substitution: G110W Q111D G112P P113W, Y114W, G112A P113W Y114S E115D, or Y114F in CDR3β compared to the parental TCR.

[0054] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(3) compared to the parental TCR: (1) An amino acid substitution Q29E or Q29P A36I T37E T38R in CDR1β; (2) Amino acid substitutions S56T N57D E58Y or S63D K64Q in CDR2β; (3) Amino acid substitutions G110W Q111D G112P P113W, Y114W, G112A P113W Y114S E115D or Y114F in CDR3β; comprises one or more of the following.

[0055] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(5) in CDR1α and / or FR1α compared to the parental TCR: (1) D27C or D27Y; (2) S28P, S28M, S28A or S28G; (3) S29Y, S29L, S29F, S29E or S29M; (4) S36L, S36Q, S36M or S36F; (5) T26V, T26S, T26I, T26P or T26Q; comprises one or more of the following, preferably, the modified TCR comprises amino acid substitutions: T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M in CDR1α and / or FR1α compared to the parental TCR.

[0056] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(4) in CDR2α compared to the parental TCR: (1) S58I or S58A; (2) N59F; (3) M63Q or M63E; (4) D64A, D64S or D64Q; comprises one or more of the following, Preferably, the modified TCR comprises, compared to the parental TCR, an amino acid substitution in CDR2α: N59F, S58IN59FM63QD64A, S58AN59FM63ED64S, S58AN59FD64Q, or S58IN59FM63QD64A.

[0057] In some embodiments, the modified TCR comprises, compared to the parental TCR, one or more of the following amino acid substitutions (1)-(2): (1) an amino acid substitution in CDR1α and / or FR1α: T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F, or T26QD27YS28GS29M; (2) an amino acid substitution in CDR2α: N59F, S58IN59FM63QD64A, S58AN59FM63ED64S, S58AN59FD64Q, or S58IN59FM63QD64A; including one or more of the above.

[0058] In some embodiments, the modified TCR (i) is a variable region of the α chain, and CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NO: 103, 76, and 29, respectively, , CDR2α, and CDR3α, CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NO: 44, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NO: 140, 141, and 29, respectively, CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NO: 146, 118, and 29, respectively, CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NO: 45, 149, and 29, respectively, CDR1α, CDR2α, and CDR3α having the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 165, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 170, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 118, and 29, respectively, or An α-chain variable region containing the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, and (ii) A β-chain variable region, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 25, and 131, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 35, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 35, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 153, and 35, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 26, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 162, respectively, or It includes a variable β-chain region containing CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:82, 91, and 33, respectively.

[0059] In some embodiments, the modified TCR has a variable α-chain region having an amino acid sequence shown in any one of SEQ ID NO:132, 136, 142, 147, 150, 154, 158, 68, 166, 119, 168, 171, 173, or an amino acid sequence selected from amino acid sequences 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 NO:132, 136, 142, 147, 150, 154, 158, 68, 166, 119, 168, 171, 173, and a variable β-chain region having an amino acid sequence shown in any one of SEQ ID NO:133, 137 143, 155, 159, 163, 175, or an amino acid sequence selected from amino acid sequences 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 NO:133, 137, 143, 155, 159, 163, 175.

[0060] In some embodiments, the modified TCR further includes 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 preferred embodiments, the TCR α-chain constant region and / or the TCR β-chain constant region includes 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 preferred embodiments, the TCR α-chain constant region includes an LVL substitution or an LIV substitution such that the α-chain constant region includes the amino acid sequence LLVIVLRIL.

[0061] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence shown in any one of SEQ ID NO: 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 NO: 134, 138, 144, 148, 151, 156, 160, 23, 167, 120, 169, 172, 174, and a TCR β-chain having an amino acid sequence shown in any one of SEQ ID NO: 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 NO: 135, 139, 145, 157, 161, 164, 176.

[0062] 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 and an MHC molecule.

[0063] In some embodiments, the MHC molecule is of the HLA-A*02 type, such as HLA-A*02:01 or HLA-A*02:05.

[0064] In some embodiments, the modified TCR is soluble or membrane-bound.

[0065] In some embodiments, the modified TCR is isolated or purified.

[0066] In another form, the present disclosure provides a modified T cell receptor (TCR) comprising a variable region of the α-chain containing CDR1α, CDR2α and CDR3α, and a variable region of the β-chain containing CDR1β, CDR2β and CDR3β, wherein compared to the parental TCR, the following amino acid substitutions are made: (i) An amino acid substitution in the variable region of the β-chain, wherein (a) an amino acid substitution at one or more positions of Gln29, Ala36, Thr37, and Thr38 in CDR1β, (b) an amino acid substitution at one or more positions of Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65 in CDR2β, and / or (c) an amino acid substitution at one or more positions of Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115 in CDR3β, and / or (ii) an amino acid substitution in the variable region of the α-chain, wherein (d) Ser28, Ser29, Ser36, Thr37, and Tyr38 in CDR1α, (e) an amino acid substitution at positions Ile56, Phe57, Ser58, Asn59, Met63, and Asp64 in CDR2α, and / or (f) an amino acid substitution at position Thr26 in FR1α, wherein the parental TCR comprises a CDR1β having the amino acid sequence shown in SEQ ID NO:24, a CDR2β having the amino acid sequence shown in SEQ ID NO:25, a CDR3β having the amino acid sequence shown in SEQ ID NO:26, a CDR1α having the amino acid sequence shown in SEQ ID NO:27, a CDR2α having the amino acid sequence shown in SEQ ID NO:28, and a CDR3α having the amino acid sequence shown in SEQ ID NO:29, wherein the amino acid positions are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT), to provide a modified TCR.

[0067] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(31) compared to the parental TCR: (1) an amino acid substitution Q29P at position Gln29 in CDR1β; (2) an 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) Amino acid substitution at position Asn57 in CDR2β selected from N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T; (7) Amino acid substitution at position Glu58 in CDR2β selected from E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W; (8) Amino acid substitution at position Gly59 in CDR2β selected from G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T; (9) Amino acid substitution at position Ser63 in CDR2β selected from S63D, S63E, S63H, S63K, S63Q, S63R, and S63T; (10) Amino acid substitution at position Lys64 in CDR2β selected from 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) Amino acid substitution at position Ser105 in CDR3β selected from S105L, S105N, S105H, S105Q, S105K, and S105R; (13) Amino acid substitution at position Glu108 in CDR3β selected from E108D, E108F, E108T, E108I, E108K, and E108P; (14) Amino acid substitution at position Gln111 in CDR3β selected from Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H; (15) Amino acid substitution of EPGQGP to VNDGT at positions from Glu108 to Pro113 in CDR3β; (16) Amino acid substitution G112E at the Gly112 position in CDR3β; (17) Amino acid substitution P113H at the Pro113 position in CDR3β; (18) Amino acid substitution at the Tyr114 position in CDR3β selected from Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S; (19) Amino acid substitution E115I at the Glu115 position in CDR3β; (20) Amino acid substitution at the Ser28 position in CDR1α selected from S28Y and S28P; (21) Amino acid substitution at the Ser29 position in CDR1α selected from S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N; (22) Amino acid substitution at the Ser36 position in CDR1α selected from S36D, S36M, S36L, S36Q, S36I, S36F, S36P, and S36V; (23) Amino acid substitution T37I at the Thr37 position in CDR1α; (24) Amino acid substitution Y38L at the Tyr38 position in CDR1α; (25) Amino acid substitution at the Ile56 position in CDR2α selected from I56T and I56G; (26) Amino acid substitution at the Phe57 position in CDR2α selected from F57W and F57T; (27) Amino acid substitution at the Ser58 position in CDR2α selected from S58G, S58I, S58N, S58A, S58P, and S58W; (28) Amino acid substitution at the Asn59 position in CDR2α selected from N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T; (29) Amino acid substitution at the Met63 position in CDR2α selected from M63D, M63Q, M63T, and M63E; (30) Amino acid substitution at the Asp64 position in CDR2α selected from D64T, D64Y, and D64S; (31) An amino acid substitution at the Thr26 position of FR1α selected from T26A and T26V; includes one or more of the following.

[0068] In some embodiments, the modified TCR does not contain the S105N amino acid substitution alone as compared to the parental TCR.

[0069] In some embodiments, the modified TCR contains one or more of the following amino acid substitutions in CDR1β as compared to the parental TCR: Q29P, A36I, T37E, and T38R. Preferably, the modified TCR contains the amino acid substitutions Q29P, A36I, T37E, and T38R in CDR1β as compared to the parental TCR.

[0070] In some embodiments, the modified TCR contains the following amino acid substitutions (1)-(7) in CDR2β as compared to the parental TCR: (1) S56T; (2) N57Q or N57D; (3) E58L, E58P, or E58Y; (4) G59Q, G59P, or G59L; (5) S63T; (6) K64C; (7) A65V; includes one or more of the following, Preferably, the modified TCR contains the amino acid substitutions N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L, or S56TN57DE58Y in CDR2β as compared to the parental TCR.

[0071] In some embodiments, the modified TCR contains the following amino acid substitutions (1)-(7) in CDR3β as compared to the parental TCR: (1) Q111G or Q111S; (2) G112E; (3) P113H; (4) Y114F, Y114L, or Y114W; (5) E115I; (6) S105L, S105Q, S105N or S105H; (7) Substitution of EPGQGP with VNDGT at positions from Glu108 to Pro113; comprising one or more of the following, preferably, the modified TCR has amino acid substitutions in CDR3β compared to the parental TCR: Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, substitution of EPGQGP with VNDGT at positions from Glu108 to Pro113 or G112EP113HY114LE115I.

[0072] In some embodiments, the modified TCR, compared to the parental 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.

[0073] In some embodiments, the modified TCR, compared to the parental TCR, has the following amino acid substitutions (1)-(6) in CDR1α and / or FR1α: (1) S28Y or S28P; (2) S29T, S29L or S29Y; (3) S36D, S36M, S36L or S36Q; (4) T37I; (5) Y38L; (6) T26A or T26V; comprising one or more of the following, preferably, the modified TCR, compared to the parental TCR, has amino acid substitutions in CDR1α and / or FR1α: S29TS36DT37IY38L, T26AS28YS29L or T26VS28PS29Y.

[0074] In some embodiments, the modified TCR, compared to the parental TCR, has the following amino acid substitutions (1)-(6) in CDR2α: (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) D64T, D64Y or D64S; comprising one or more of: Preferably, the modified TCR comprises, compared to the parental TCR, amino acid substitutions S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F in CDR2α.

[0075] In some embodiments, the modified TCR comprises: (i) a variable α-chain region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27-29, respectively, and (ii) a variable β-chain region, wherein 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 NO:24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 88, and 26, respectively, the CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 26, respectively, or a β-chain variable region comprising the CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 94, respectively.

[0076] In some embodiments, the modified TCR (i) a β-chain variable region comprising the CDR1β, C DR2β, and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24-26, respectively, and (ii) an α-chain variable region, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 42, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 43, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 44, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 45, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 76, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 97, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 100, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 103, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27, 106, and 29, respectively, the 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 NO: 27, 112, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, or an α-chain variable region comprising CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively.

[0077] In some embodiments, the modified TCR is (i) an α-chain variable region comprising CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 42, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 97, 28, and 29, respectively, CDR1α of the amino acid sequences shown in SEQ ID NO: 100, 28, and 29, respectively , CDR2α, and CDR3α, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 106, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 109, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, or an α-chain variable region containing CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively, and (ii) a β-chain variable region, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 31, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 32, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 88, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 26, respectively, or a β-chain variable region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 94, respectively.

[0078] In some embodiments, the modified TCR comprises an α-chain variable region having the amino acid sequence 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 β-chain variable region having the amino acid sequence selected from the amino acid sequences shown 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.

[0079] In some embodiments, the modified TCR comprises an α-chain variable region having the amino acid sequence selected from the amino acid sequences shown in 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: 65 - 68, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, and a β-chain variable region having the amino acid sequence shown 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.

[0080] In some embodiments, the modified TCR has an α-chain variable region having an amino acid sequence shown in 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 selected from amino acid sequences 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 a β-chain variable region having an amino acid sequence shown in any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, or an amino acid sequence selected from amino acid sequences 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.

[0081] 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 comprises 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.

[0082] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence 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 β-chain having an amino acid sequence selected from the amino acid sequences 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.

[0083] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in 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 a TCR β-chain having an amino acid sequence shown 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.

[0084] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in 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 a TCR β-chain having an amino acid sequence selected from the amino acid sequences 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.

[0085] 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 and an MHC molecule.

[0086] In some embodiments, the MHC molecule is of the HLA-A*02 type, such as HLA-A*02:01 or HLA-A*02:05.

[0087] In some embodiments, the modified TCR is soluble or membrane-bound.

[0088] In some embodiments, the modified TCR is isolated or purified.

[0089] In yet another aspect, the present disclosure provides a modified T cell receptor (TCR) comprising a variable α-chain region comprising CDR1α, CDR2α and CDR3α, and a variable β-chain region comprising CDR1β, CDR2β and CDR3β, wherein compared to the parental TCR, the following amino acid substitutions are made: (i) An amino acid substitution in the variable region of the β-chain, wherein (a) an amino acid substitution at one or more positions of Gln29, Ala36, Thr37, Thr38 in CDR1β, (b) an amino acid substitution at one or more positions of Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, Ala65 in CDR2β, and / or (c) an amino acid substitution at one or more positions of Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114 and Glu115 in CDR3β, and / or (ii) an amino acid substitution in the variable region of the α-chain, wherein (d) an amino acid substitution at one or more positions of Asp27, Ser28, Ser29, Ser36, Thr37 and Tyr38 in CDR1α, (e) an amino acid substitution at positions of Ile56, Phe57, Ser58, Asn59, Met63 and Asp64 in CDR2α, and / or (f) an amino acid substitution at position Thr26 in FR1α, wherein the parent TCR comprises a CDR1β having the amino acid sequence shown in SEQ ID NO:24, a CDR2β having the amino acid sequence shown in SEQ ID NO:25, a CDR3β having the amino acid sequence shown in SEQ ID NO:26, a CDR1α having the amino acid sequence shown in SEQ ID NO:27, a CDR2α having the amino acid sequence shown in SEQ ID NO:28, and a CDR3α having the amino acid sequence shown in SEQ ID NO:29; wherein the amino acid positions are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT), providing a modified TCR.

[0090] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(33) compared to the parent TCR: (1) An amino acid substitution at position Gln29 in CDR1β selected from 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) Amino acid substitution at position Asn57 in CDR2β selected from N57D, N57E, N57H, N57K, N57Q, N57R, N57S and N57T; (7) Amino acid substitution at position Glu58 in CDR2β selected from E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y and E58W; (8) Amino acid substitution at position Gly59 in CDR2β selected from G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S and G59T; (9) Amino acid substitution at position Ser63 in CDR2β selected from S63D, S63E, S63H, S63K, S63Q, S63R and S63T; (10) Amino acid substitution at position Lys64 in CDR2β selected from 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) Amino acid substitution at position Ser105 in CDR3β selected from S105L, S105N, S105H, S105Q, S105K and S105R; (13) Amino acid substitution at position Glu108 in CDR3β selected from E108D, E108F, E108T, E108I, E108K and E108P; (14) Amino acid substitution G110W at position Gly110 in CDR3β; (15) Amino acid substitution at position Gln111 in CDR3β selected from Q111D, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L and Q111H; (16) Amino acid substitution of EPGQGP to VNDGT at positions from Glu108 to Pro113 in CDR3β; (17) Amino acid substitution at position Gly112 in CDR3β, selected from G112P, G112A and G112E; (18) Amino acid substitution at position Pro113 in CDR3β, selected from P113W and P113H; (19) Amino acid substitution at position Tyr114 in CDR3β, selected from Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P and Y114S; (20) Amino acid substitution at position Glu115 in CDR3β, selected from E115D and E115I; (21) Amino acid substitution at position Asp27 in CDR1α, selected from D27C and D27Y; (22) Amino acid substitution at position Ser28 in CDR1α, selected from S28M, S28A, S28G, S28Y and S28P; (23) Amino acid substitution at position Ser29 in CDR1α, selected from S29E, S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W and S29N; (24) Amino acid substitution at position Ser36 in CDR1α, selected from 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) Amino acid substitution at position Ile56 in CDR2α, selected from I56T and I56G; (28) Amino acid substitution at position Phe57 in CDR2α, selected from F57W and F57T; (29) Amino acid substitution at position Ser58 in CDR2α, selected from S58G, S58I, S58N, S58A, S58P and S58W; (30) An amino acid substitution at the Asn59 position in CDR2α selected from N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T; (31) An amino acid substitution at the Met63 position in CDR2α selected from M63D, M63Q, M63T, and M63E; (32) An amino acid substitution at the Asp64 position in CDR2α selected from D64A, D64Q, D64T, D64Y, and D64S; (33) An amino acid substitution at the Thr26 position in FR1T selected from T26S, T26I, T26P, T26Q, T26A, and T26V; Comprising one or more of the above.

[0091] In some embodiments, the modified TCR does not contain a single S10 5N amino acid substitution compared to the parental TCR.

[0092] In some embodiments, the modified TCR contains one or more of the following amino acid substitutions (1)-(4) in CDR1β compared to the parental TCR: (1) Q29E or Q29P; (2) A36I; (3) T37E; (4) T38R; Comprising one or more of the above, Preferably, the modified TCR contains the amino acid substitution Q29E or Q29P A36I T37E T38R in CDR1β compared to the parental TCR.

[0093] In some embodiments, the modified TCR contains one or more of the following amino acid substitutions (1)-(7) in CDR2β compared to the parental 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; comprising one or more of: Preferably, the modified TCR comprises, compared to the parental TCR, the amino acid substitutions N57D E58Y G59Q S63T K64C A65V, S56T E58L G59P, S56T N57Q E58P G59L, S56T N57D E58Y or S63D K64Q in CDR2β.

[0094] In some embodiments, the modified TCR comprises, compared to the parental TCR, the following amino acid substitutions (1)-(8) in CDR3β: (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) Substitution of EPGQGP with VNDGT at positions from Glu108 to Pro113; comprising one or more of: Preferably, the modified TCR comprises, compared to the parental TCR, the amino acid substitutions G110W Q111D G112P P113W, G112A P113W Y114S E115D, Y114F, Y114W, Q111G Y114F, Q111S Y114F, S105L, S105N, substitution of EPGQGP with VNDGT at positions from Glu108 to Pro113 or G112E P113H Y114L E115I in CDR3β.

[0095] In some embodiments, the modified TCR comprises, compared to the parental TCR, the following amino acid substitutions (1)-(3): (1) Amino acid substitution Q29E or Q29P A36I T37E T38R in CDR1β; (2) Amino acid substitutions in CDR2β: N57D, E58Y, G59Q, S63T, K64C, A65V; S56T, E58L, G59P; S56T, N57Q, E58P, G59L; S56T, N57D, E58Y or S63D, K64Q; 58Y or S63D, K64Q; (3) Amino acid substitutions in CDR3β: G110W, Q111D, G112P, P113W; G112A, P113W, Y114S, E115D; Y114F; Y114W; Q111G, Y114F; Q111S, Y114F; S105L; S105N; substitution of EPGQGP with VNDGT at positions from Glu108 to Pro113 or G112E, P113H, Y114L, E115I; comprising one or more of the above.

[0096] In some embodiments, the modified TCR comprises amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V in CDR2β as compared to the parental TCR; and amino acid substitutions Y114F, Y114L, Y114W, Q111G, Y114F or Q111S, Y114F in CDR3β.

[0097] In some embodiments, the modified TCR comprises one or more of the following amino acid substitutions (1)-(7) in CDR1α and / or FR1α as compared to the parental TCR: (1) D27C or D27Y; (2) S28Y, S28P, S28M, S28A or S28G; (3) S29T, S29L, S28Y, S29F, S28E or S29M; (4) S36F, S36D, S36M, S36L or S36Q; (5) T37I; (6) Y38L; (7) T26A, T26V, T26S, T26I, T26P or T26Q; comprising one or more of the above, Preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M.

[0098] In some embodiments, compared with the parental TCR, the modified TCR comprises the following amino acid substitutions (1)-(6) in CDR2α: (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; including one or more of: Preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR2α: N59F, S58I N59F M63Q D64A, S58A N59F D64Q, S58I N59F M63Q D64A, S58G N59F M63D D64T, S58I N59F M63Q D64Y, S58N N59T M63Q, N59F M63T D64Y, S58A N59F M63E D64S, I56T F57W S58P N59H, I56G F57T S58W N59L or I56T S58W N59F.

[0099] In some embodiments, compared with the parental TCR, the modified TCR comprises the following amino acid substitutions (1)-(2): (1) Amino acid substitutions contained in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28 MS29F, S36M, S29ES36F or T26QD27YS28GS29M; (2) Amino acid substitutions contained in CDR2α: N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F; comprising one or more of the above.

[0100] In some embodiments, the modified TCR is (i) an α-chain variable region comprising CDR1α, CDR2α and CDR3α with the amino acid sequences shown in SEQ ID NOs: 27-29 respectively, and (ii) a β-chain variable region, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26 respectively, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31 respectively, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32 respectively, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33 respectively, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34 respectively, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35 respectively, CDR1β, CDR2β and CDR3β with the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33 respectively, CDR1β, CDR2β and CDR3β with 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 NO:24, 30 and 35 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 36 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 37 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 36 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 37 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 38 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 39 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 40 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 41 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 79 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:82, 25 and 26 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 85 and 26 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 88 and 26 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 91 and 26 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 94 respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 25, and 131, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 153, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 162, respectively, or a β-chain variable region containing CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 91, and 33, respectively.

[0101] In some embodiments, the modified TCR (i) a β-chain variable region containing CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24 to 26, respectively, (ii) an α-chain variable region, 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 NO: 97, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 100, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 106, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 109, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29, respectively, CDR1α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29, respectively , CDR2α, and CDR3α, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 140, 141, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 146, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:45, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:152, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:43, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:45, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:165, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:27, 118, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:45, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:170, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:43, 118, and 29, respectively, or An α-chain variable region comprising the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:152, 149, and 29, respectively.

[0102] In some embodiments, the modified TCR is (i) An α-chain variable region, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:42, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:43, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:44, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 97, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 100, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 106, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 109, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 140, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 146, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 165, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 170, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 118, and 29, respectively, or An α-chain variable region containing the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, and (ii) A β-chain variable region, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 26, respectively, The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 31, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 32, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 88, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 94, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 25, and 131, respectively, CDR1β of the amino acid sequences shown in SEQ ID NO:24, 91, and 35, respectively, CDR2β and CDR3β, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 153, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 91, and 162, respectively, or a β-chain variable region containing CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:82, 91, and 33, respectively.

[0103] In some embodiments, the modified TCR comprises an α-chain variable region having the amino acid sequence 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 β-chain variable region having an amino acid sequence selected from the amino acid sequences shown 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.

[0104] In some embodiments, the modified TCR comprises an α-chain variable region having the amino acid sequence shown in 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 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, 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 the amino acid sequence shown 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.

[0105] In some embodiments, the modified TCR has an α-chain variable region having an amino acid sequence shown in 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 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, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, and a β-chain variable region having an amino acid sequence shown 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, and is selected from the amino acid sequences thus selected. It includes a β-chain variable region having an amino acid sequence selected.

[0106] In some embodiments, the modified TCR further includes 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 includes at least one cysteine substitution with respect to the wild-type sequence in order 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 includes an LVL substitution or an LIV substitution such that the α-chain constant region includes the amino acid sequence LLVIVLRIL.

[0107] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence 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 β-chain having an amino acid sequence selected from the amino acid sequences 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.

[0108] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in 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 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, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174, and a TCR β-chain having an amino acid sequence shown 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.

[0109] In some embodiments, the modified TCR comprises a TCR α-chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 20 to 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 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 to 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 β-chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 3 to 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 to 19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176.

[0110] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2), or a complex of said epitope and an MHC molecule.

[0111] In some embodiments, the MHC molecule is of the HLA-A*02 type, such as HLA-A*02:01 or HLA-A*02:05.

[0112] In some embodiments, the modified TCR is soluble or membrane-bound.

[0113] In some embodiments, the modified TCR is isolated or purified.

[0114] In another form, the present disclosure provides a nucleic acid encoding a modified TCR of the present invention.

[0115] In yet another form, the present disclosure provides a vector comprising the nucleic acid of the present invention.

[0116] In some embodiments, the vector is selected from a lentiviral vector, a retroviral vector, a plasmid, a DNA vector, an mRNA vector, a transposon-based vector, and an artificial chromosome.

[0117] In another form, the present disclosure provides a cell comprising a modified TCR, nucleic acid, or vector of the present invention.

[0118] In some embodiments, the cell is selected from lymphocytes (e.g., T cells, NK cells), monocytes (e.g., PBMC), and stem cells. In some embodiments, the stem cells are lymphocyte progenitor cells or induced pluripotent stem cells (iPSCs). In some embodiments, the cell is a T cell. In some embodiments, the T cell does not express an endogenous TCR.

[0119] In yet another form, the present disclosure provides a method for producing a cell of the present invention, comprising the step of transducing or transfecting a cell with a vector of the present invention.

[0120] In some embodiments, the method further comprises the step of growing and / or activating the cell before or after the transduction or transfection.

[0121] In another form, the present disclosure provides a conjugate comprising a modified TCR according to the present invention and an active agent bound or conjugated to the TCR.

[0122] In some embodiments, the active agent is selected from a detectable label, an immune-stimulating 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 immune-stimulating molecule is selected from cytokines, chemokines, platelet factors and complement promoters; preferably, the therapeutic agent is selected from immunomodulators, radioactive compounds, enzymes, chemotherapeutic agents and toxins.

[0123] In yet another aspect, the disclosure provides a bispecific or multispecific antigen-binding molecule comprising a modified TCR of the invention or an antigen-binding fragment thereof and one or more additional antigen-binding regions that bind to one or more additional antigens, optionally, the one or more additional antigen-binding regions are antigen-binding regions derived from an antibody and antigen-binding regions derived from a TCR and are each independently selected therefrom.

[0124] In some embodiments, the one or more additional antigens are each independently selected from tumor-associated antigens (TAAs), tumor-specific antigens (TSAs), viral antigens, self-antigens, and immune cell surface molecules. In some embodiments, the one or more additional antigens are CD3, preferably, the one or more additional antigen-binding regions are derived from the antigen-binding region of an anti-CD3 antibody.

[0125] In another aspect, the disclosure provides a composition comprising a modified TCR, nucleic acid, vector, cell, conjugate, or bispecific or multispecific antigen-binding molecule according to the invention, preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient.

[0126] In some embodiments, the composition further comprises a second therapeutic agent, preferably, the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs.

[0127] In yet another aspect, the present disclosure provides a method of treating MAGE-A1 positive cancer in a subject, comprising administering to the subject an effective amount of the modified TCR of the present invention.

[0128] In another aspect, the present disclosure provides a method of treating MAGE-A1 positive cancer in a subject, comprising administering to the subject an effective amount of the cells of the present invention.

[0129] In some embodiments of the treatment methods of the present disclosure, the cells are autologous or allogeneic to the subject.

[0130] In some embodiments, the method comprises (i) isolating a cell-containing sample from the subject; (ii) transducing or transfecting the cells with the vector of the present invention; and (iii) administering the cells obtained in step (ii) to the subject.

[0131] In some embodiments, the method further comprises knocking out the endogenous TCR in the cells after step (i) and before step (ii).

[0132] 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 carcinoma of the bladder), ovarian cancer (e.g., serous cystadenocarcinoma of the ovary), head and neck cancer (e.g., squamous cell carcinoma of the head and neck), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[0133] In some embodiments, the method further comprises administering a second therapeutic agent, preferably, the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs.

[0134] In some embodiments of the treatment methods of the present disclosure, the subject has an HLA-A*02 allele, such as an HLA-A*02:01 or HLA-A*02:05 allele.

[0135] In another form, the present disclosure provides the use of the modified TCR, nucleic acid, vector, cell, conjugate, bispecific or multispecific antigen-binding molecule or composition of the present invention in the manufacture of a medicament for the treatment of MAGE-A1 positive cancer in a subject.

[0136] In yet another form, the present disclosure provides the modified TCR, nucleic acid, vector, cell, conjugate, bispecific or multispecific antigen-binding molecule or composition of the present invention for use in the treatment of MAGE-A1 positive cancer in a subject.

[0137] 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., serous cystadenocarcinoma of the ovary), head and neck cancer (e.g., squamous cell carcinoma of the head and neck), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma. In some embodiments, the subject has an HLA-A*02:01 allele, such as an HLA-A*02:01 or HLA-A*02:05 allele. BRIEF DESCRIPTION OF THE DRAWINGS

[0138]

Figure 1-1

Figure 1-2

Figure 1-3

Figure 1-4

Figure 1-5

Figure 1-6

Figure 2-1

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Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9-1

Figure 9-2

Figure 10

Figure 11-1

Figure 11-2

Figure 12

BEST MODE FOR CARRYING OUT THE INVENTION

[0139] Unless otherwise indicated, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art. For example, the terms used in this specification are as defined 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)", edited by Leuenberger, H.G.W, Nagel, B. and Kolbl, H. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland.

[0140] It should be noted that, as used in this specification and 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.

[0141] The term "comprising", as used in this specification and the appended claims, 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". In the following, when a group is defined as including or comprising at least some embodiments, it should preferably be understood that a group consisting of only these embodiments is disclosed.

[0142] In one embodiment, the present disclosure provides a modified T cell receptor (TCR) comprising a variable α-chain region comprising CDR1α, CDR2α, and CDR3α, and a variable β-chain region comprising CDR1β, CDR2β, and CDR3β, wherein, compared to a parental TCR, the following amino acid substitutions are made: (i) an amino acid substitution in the variable β-chain region, wherein (a) an amino acid substitution at one or more positions of Gln29, Ala36, Thr37, and Thr38 in CDR1β, (b) an amino acid substitution at one or more positions of Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65 in CDR2β, and / or (c) an amino acid substitution at one or more positions of Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115 in CDR3β, and / or (ii) an amino acid substitution in the variable α-chain region, wherein (d) an amino acid substitution at one or more positions of Asp27, Ser28, Ser29, Ser36, Thr37, and Tyr38 in CDR1α, (e) an amino acid substitution at positions Ile56, Phe57, Ser58, Asn59, Met63, and Asp64 in CDR2α, and / or (f) an amino acid substitution at position Thr26 in FR1α, wherein the parental TCR comprises a CDR1β having the amino acid sequence shown in SEQ ID NO:24, a CDR2β having the amino acid sequence shown in SEQ ID NO:25, a CDR3β having the amino acid sequence shown in SEQ ID NO:26, a CDR1α having the amino acid sequence shown in SEQ ID NO:27, a CDR2α having the amino acid sequence shown in SEQ ID NO:28, and a CDR3α having the amino acid sequence shown in SEQ ID NO:29, wherein the amino acid positions are numbered according to the nomenclature of the International ImMunoGeneTics information system (IMGT), providing a modified TCR.

[0143] In some embodiments, the modified TCR has the following amino acid substitutions compared to the parental TCR: (i) An amino acid substitution in the variable region of the β-chain, wherein (a) the amino acid substitution at one or more positions of Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65 in CDR2β, and / or (b) the amino acid substitution at one or more positions of Ser105, Glu108, Gln111 and Tyr114 in CDR3β, and and / or (ii) An amino acid substitution in the variable region of the α-chain, wherein (c) the amino acid substitution at positions Ser29 and Ser36 in CDR1α, and / or (d) the amino acid substitution at position Asn59 in CDR2α, and wherein the parental TCR comprises a CDR1β having the amino acid sequence shown in SEQ ID NO:24, a CDR2β having the amino acid sequence shown in SEQ ID NO:25, a CDR3β having the amino acid sequence shown in SEQ ID NO:26, a CDR1α having the amino acid sequence shown in SEQ ID NO:27, a CDR2α having the amino acid sequence shown in SEQ ID NO:28, and a CDR3α having the amino acid sequence shown in SEQ ID NO:29; wherein the amino acid positions are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT).

[0144] In its native form, the TCR exists as a complex of several proteins on the surface of T cells. The T cell receptor is produced by the independent T cell receptor α and β (TCRα and TCRβ) genes and consists of two (separate) protein chains called the α-chain and the β-chain. Each chain of the TCR has one N-terminal immunoglobulin-like (Ig) variable (V) region / domain, one Ig constant (C) region / domain, a transmembrane / cell membrane spanning region that anchors the chain to the cell membrane, and a short cytoplasmic tail at the C-terminus.

[0145] Generally, the TCR is a heterodimer of an α-chain and a β-chain that can bind to MHC molecules presenting antigen peptides.

[0146] Antigen specificity is conferred by the variable regions of the α-chain and the β-chain. The two variable regions of the TCR α-chain and β-chain (α-chain variable region (Vα) and β-chain variable region (Vβ)) each 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 major determinant of antigen recognition and specificity (i.e., the ability to recognize and interact with a specific antigen), while CDR1 and CDR2 mainly interact with MHC molecules presenting antigen peptides.

[0147] The TCR recognizes antigen peptides (presented / displayed on MHC molecules) that bind to major histocompatibility complex (MHC) molecules on the surface of antigen-presenting cells. Antigen peptides presented on MHC molecules are also referred to herein as "epitope-MHC complexes", "epitope-MHC conjugates", or "target antigen peptide-MHC complexes". There are two different classes of MHC molecules, MHC class I and MHC class II, which present peptides from different cellular compartments. MHC class I molecules are expressed on the surface of all nucleated cells in humans and display peptides or protein fragments from intracellular compartments to cytotoxic T cells. In humans, MHC is also called human leukocyte antigen (HLA). MHC class I has three major types: HLA-A, HLA-B, and HLA-C. When the TCR binds to its specific epitope-MHC complex, the T cell is activated and exerts biological effector functions.

[0148] The positions of the amino acid sequences of TCRs in the present invention are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT). For example, the position numbers of the amino acid sequences of the variable region of the TCR α-chain (Vα) and the variable region of the β-chain (Vβ) follow the positions listed in IMGT. For example, if a certain amino acid in Vα has a position number of 29 listed in IMGT, it is described in this specification as the 29th amino acid of Vα; if a certain amino acid in Vβ has a position number of 57 listed in IMGT, it is described in this specification as the 57th amino acid of Vβ; the same applies to the others. Here, when the position numbers of the sequences of other amino acids are to be specifically explained, they will be explained accordingly.

[0149] As used herein, the term "parent TCR" refers to the previously developed HLA-A0201-restricted human-derived TCR ht27 that targets MAGE-A1 and includes CDR1β shown by the amino acid sequence SEQ ID NO :24 (DFQATT), CDR2β shown by the amino acid sequence SEQ ID NO:25 (SNEGSKA), CDR3β shown by the amino acid sequence SEQ ID NO:26 (SAREPGQGPYEQY), CDR1α shown by the amino acid sequence SEQ ID NO:27 (DSSSTY), CDR2α shown by the amino acid sequence SEQ ID NO:28 (IFSNMDM), and CDR3α shown by the amino acid sequence SEQ ID NO:29 (AGSGGGTDKLI), and the amino acid sequences of the variable region of the α-chain and the variable region of the β-chain are shown by SEQ ID NO:46 and SEQ ID NO:47, respectively.

[0150] Variable region of the α-chain: RGEDVEQSLFLSVREGDSSVINCTYTDSSSTYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAGSGGGTDKLIFGTGTRLQVFPN (SEQ ID NO:46)

[0151] Variable region of the β-chain: GAVVSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLMATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSAREPGQGPYEQYFGPGTRLTVTE(SEQ ID NO:47).

[0152] The inventors of the present invention have found that the modified TCRs of the present invention, which contain one or more amino acid substitutions at the following positions in the variable region compared to the parental TCR: Asn57, Glu58, Gly59, Ser63, Lys64, Ala65 in CDR2β, Ser105, Glu108, Gln111, Tyr114 in CDR3β, Ser29 and Ser36 in CDR1α, and Asn59 in CDR2α, have improved binding affinity for MAGE-A1. For example, an amino acid substitution at the Ser105 position can increase the binding affinity of the TCR by about 10 to 400-fold; an amino acid substitution at the Glu108 position can increase the binding affinity of the TCR by about 1 to 2-fold; an amino acid substitution at the Gln111 position can increase the binding affinity of the TCR by about 12 to 14-fold; an amino acid substitution at the Tyr114 position can increase the binding affinity of the TCR by about 11 to 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, G59T, 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, E108T, E108I, E108K, E108P, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, Q111H, Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, 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.

[0153] Thus, in some embodiments, the modified TCR has the following amino acid substitutions (1)-(33) compared to the parental TCR: (1) An amino acid substitution at position Gln29 in CDR1β selected from Q29E and Q29P; (2) The amino acid substitution A36I at position Ala36 in CDR1β; (3) The amino acid substitution T37E at position Thr37 in CDR1β; (4) The amino acid substitution T38R at position Thr38 in CDR1β; (5) The amino acid substitution S56T at position Ser56 in CDR2β; (6) An amino acid substitution at position Asn57 in CDR2β selected from N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T; (7) An amino acid substitution at position Glu58 in CDR2β selected from E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W; (8) An amino acid substitution at position Gly59 in CDR2β selected from G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T; (9) An amino acid substitution at position Ser63 in CDR2β selected from S63D, S63E, S63H, S63K, S63Q, S63R, and S63T; (10) An amino acid substitution at position Lys64 in CDR2β selected from K64Q, K64M, K64C, K64W, K64T, K64P, and K64A; (11) An 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 S105L, S105N, S105H, S105Q, S105K, and S105R; (13) An amino acid substitution at position Glu108 in CDR3β selected from E108D, E108F, E108T, E108I, E108K, and E108P; (14) Amino acid substitution G110W at the Gly110 position in CDR3β; (15) Amino acid substitution at the Gln111 position in CDR3β, selected from Q111D, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H; (16) Amino acid substitution of EPGQGP to VNDGT at the positions from Glu108 to Pro113 in CDR3β; (17) Amino acid substitution at the Gly112 position in CDR3β, selected from G112P, G112A, and G112E; (18) Amino acid substitution at the Pro113 position in CDR3β, selected from P113W and P113H; (19) Amino acid substitution at the Tyr114 position in CDR3β, selected from Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S; (20) Amino acid substitution at the Glu115 position in CDR3β, selected from E115D and E115I; (21) Amino acid substitution at the Asp27 position in CDR1α, selected from D27C and D27Y; (22) Amino acid substitution at the Ser28 position in CDR1α, selected from S28M, S28A, S28G, S28Y, and S28P; (23) Amino acid substitution at the Ser29 position in CDR1α, selected from S29E, S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N; (24) Amino acid substitution at the Ser36 position in CDR1α, selected from S36D, S36M, S36L, S36Q, S36I, S36F, S36P, and S36V; (25) Amino acid substitution T37I at the Thr37 position in CDR1α; (26) Amino acid substitution Y38L at the Tyr38 position in CDR1α; (27) Amino acid substitution at the Ile56 position in CDR2α, selected from I56T and I56G; acid substitution; (28)An amino acid substitution at the Phe57 position in CDR2α selected from F57W and F57T; (29)An amino acid substitution at the Ser58 position in CDR2α selected from S58G, S58I, S58N, S58A, S58P and S58W; (30)An amino acid substitution at the Asn59 position in CDR2α selected from N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S and N59T; (31)An amino acid substitution at the Met63 position in CDR2α selected from M63D, M63Q, M63T and M63E; (32)An amino acid substitution at the Asp64 position in CDR2α selected from D64A, D64Q, D64T, D64Y and D64S; (33)An amino acid substitution at the Thr26 position in FR1α selected from T26S, T26I, T26P, T26Q, T64A and T26V; Comprising one or more of the above.

[0154] In some embodiments, the modified TCR has the following amino acid substitutions (1)-(13) compared to the parental TCR: (1)An amino acid substitution at the Asn57 position selected from N57D, N57E, N57H, N57K, N57Q, N57R, N57S and N57T; (2)An amino acid substitution at the Glu58 position selected from E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y and E58W; (3)An amino acid substitution at the Gly59 position selected from G59D, G59E, G59H, G59K, G59Q, G59R, G59S and G59T; (4)An amino acid substitution at the Ser63 position selected from S63D, S63E, S63H, S63K, S63Q, S63R and S63T; (5)An amino acid substitution at the Lys64 position selected from K64M, K64C, K64W, K64T, K64P and K64A; (6)An amino acid substitution at the AlA65 position selected from A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y; (7)An amino acid substitution at the Ser105 position selected from S105L, S105N, S105H, S105Q, S105K, and S105R; (8)An amino acid substitution at the Glu108 position selected from E108D, E108F, E108T, E108I, E108K, and E108P; (9)An amino acid substitution at the Gln111 position selected from Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H; (10)An amino acid substitution at the Tyr114 position selected from Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S; (11)An amino acid substitution at the Ser29 position selected from S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, and S29N; (12)An amino acid substitution at the Ser36 position selected from S36M, S36L, S36Q, S36I, S36F, S36P, and S36V; (13)An amino acid substitution at the Asn59 position selected from N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S, and N59T; Comprising one or more of the above.

[0155] In some embodiments, the modified TCR does not contain the S105N amino acid substitution alone compared to the parental TCR. In some embodiments, the modified TC R contains a combination of the amino acid substitution S105N and additional amino acid substitutions listed above compared to the parental TCR.

[0156] In some embodiments, the modified TCR, compared to the parental TCR, comprises one or more of the following amino acid substitutions in CDR2β: N57D, E58Y, G59Q, S63T, K64C, and A65V. In a preferred embodiment, the modified TCR, compared to the parental TCR, comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β. In some embodiments, the modified TCR, compared to the parental TCR, 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. In some embodiments, the modified TCR, compared to the parental TCR, comprises the amino acid substitutions Q111GY114F, Q111GY114L, Q111GY114W, Q111SY114F, Q111SY114L, or Q114SY114W in CDR3β. In a preferred embodiment, the modified TCR, compared to the parental TCR, comprises the amino acid substitutions Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, or S105N in CDR3β.

[0157] In some embodiments, the modified TCR, compared to the parental TCR, comprises 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 in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L, or Y114W; (3) S105L, S105Q, S105N, or S105H.

[0158] In some embodiments, the modified TCR, compared to the parental TCR, includes 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.

[0159] In some embodiments, the modified TCR, compared to the parental TCR, includes amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β, and includes amino acid substitutions Q111GY114F, Q111GY114L, Q111GY114W, Q111SY114F, Q111SY114L, or Q114SY114W in CDR3β.

[0160] In some embodiments, the modified TCR, compared to the parental TCR, includes amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β, and includes amino acid substitutions Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, or S105N in CDR3β.

[0161] In some embodiments, the modified TCR, compared to the parental TCR, includes amino acid substitutions N57D, E58Y, G59Q, S63T, K64C, and A65V in CDR2β, and includes amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F, or Q111SY114F in CDR3β. In some embodiments, the modified TCR, compared to the parental TCR, includes amino acid substitutions (1) and / or (2) below in CDR1α: (1) S29L; (2) S36M, S36L, or S36Q.

[0162] In some embodiments, the modified TCR, compared to the parental TCR, includes amino acid substitution N59F in CDR2α.

[0163] In some embodiments, the modified TCR comprises, compared to the parental TCR, the following amino acid substitutions (1) and / or (2) in CDR1α: (1) S29L; (2) S36M, S36L or S36Q, and comprises the amino acid substitution N59F in CDR2α.

[0164] Each of the three CDR regions of the TCR α and β chains binds to different sites of the MHC-polypeptide complex and acts on the MHC-polypeptide complex in a relatively independent manner. The mutations in the CDR regions disclosed herein, whether single mutations or combined mutations, effectively improve the affinity of the TCR compared to the wild-type TCR. Depending on the combination of mutations, a synergistic effect that achieves a higher affinity can also be brought about.

[0165] Thus, in some embodiments, the modified TCR comprises, compared to the parental TCR, the following amino acid substitutions (1) and / or (2) in CDR1α: (1) S29L; (2) S36M, S36L or S36Q, and comprises the amino acid substitution N59F in CDR2α, and comprises the following amino acid substitutions in CDR2β: one or more of N57D, E58Y, G59Q, S63T, K64C and A65V, and comprises the following amino acid substitutions (1)-(3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L or Y114W; (3) S105L, S105Q, S105N or S105H. In some embodiments, the modified TCR comprises, compared to the parental TCR, the following amino acid substitutions (1) and / or (2) in CDR1α: (1) S29L; (2) S36M, S36L or S36Q, comprises the amino acid substitution N59F in CDR2α, and comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V in CDR2β, and comprises the following amino acid substitutions (1)-(3) in CDR3β: (1) Q111G or Q111S; (2) Y114F, Y114L or Y114W; (3) S105L, S105Q, S105N or S105H.

[0166] In some embodiments, the modified TCR, compared to the parental TCR, comprises in CDR1α the amino acid substitutions of (1) and / or (2) below: (1) S29L; (2) S36M, S36L or S36Q, in CDR2α comprises the amino acid substitution N59F, and in CDR2β comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V, and in CDR3β comprises the amino acid substitutions Q111GY114F, Q111GY114L, Q111GY114W, Q111SY114F, Q111SY114L or Q114SY114W.

[0167] In some embodiments, the modified TCR, compared to the parental TCR, comprises in CDR1α the amino acid substitutions of (1) and / or (2) below: (1) S29L; (2) S36M, S36L or S36Q, in CDR2α comprises the amino acid substitution N59F, and in CDR2β comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V, and in CDR3β comprises the amino acid substitutions Y114F, Y114W, Q111GY114F, Q111SY114F, S105L or S105N.

[0168] In some embodiments, the modified TCR, compared to the parental TCR, comprises in CDR1α the amino acid substitutions of (1) and / or (2) below: (1) S29L; (2) S36M, S36L or S36Q, in CDR2α comprises the amino acid substitution N59F, and in CDR2β comprises the amino acid substitutions N57D, E58Y, G59Q, S63T, K64C and A65V, and in CDR3β comprises the amino acid substitutions Y114F, Y114L, Y114W, Q111GY114F or Q111SY114F.

[0169] In some embodiments, the modified TCR (i) an α-chain variable region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NOs: 27-29, respectively, and (ii) a β-chain variable region, wherein CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 26 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 31 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 32 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 33 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 34 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 35 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 33 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 34 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 35 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 36 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 37 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 36 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 30 and 37 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 38 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25 and 39 respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 88, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 26, respectively, CDR1β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 94, respectively, CDR2β and CDR3β, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 25, and 131, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 153, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 162, respectively, or CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 91, and 33, respectively, and includes a variable region of the β-chain.

[0170] In some embodiments, the modified TCR comprises (i) a variable α-chain region comprising CDR1α, CDR2α and CDR3α having the amino acid sequences shown in SEQ ID NOs: 27-29, respectively, (ii) a variable β-chain region, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30 and 26, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 31, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 32, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 33, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 34, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 35, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30 and 33, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30 and 34, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30 and 35, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 36, respectively, CDR1β, CDR2β and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25 and 37, respectively, CDR1β, CDR2β and CDR3β having 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 NO:24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 40, respectively, or a β-chain variable region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 41, respectively.

[0171] 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: 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.

[0172] 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: 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, 25, and 31, respectively.

[0173] 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: 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, 25, and 32, respectively.

[0174] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 33, respectively.

[0175] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 34, respectively.

[0176] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 35, respectively.

[0177] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30, and 33, respectively.

[0178] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30, and 34, respectively.

[0179] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30, and 35, respectively.

[0180] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 36, respectively.

[0181] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 37, respectively.

[0182] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30, and 36, respectively.

[0183] In a preferred embodiment, 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 to 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 30, and 37, respectively.

[0184] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 38, respectively.

[0185] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 39, respectively.

[0186] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 40, respectively.

[0187] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 41, respectively.

[0188] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 79, respectively.

[0189] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 82, 25, and 26, respectively.

[0190] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 85, and 26, respectively.

[0191] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 88, and 26, respectively.

[0192] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 91, and 26, respectively.

[0193] In a preferred embodiment, 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 having the amino acid sequences shown in SEQ ID NOs: 24, 25, and 94, respectively.

[0194] In some embodiments, the modified TCR is (i) a β-chain variable region comprising CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NOs: 24 to 26 respectively, (ii) an α-chain variable region, 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, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:27, 121, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:27, 124, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:27, 127, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:103, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:44, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:140, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:146, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:45, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:152, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:43, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:45, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:165, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:27, 118, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:45, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO:170, 76, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 118, and 29, respectively, or an α-chain variable region comprising the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively.

[0195] In some embodiments, the modified TCR (i) a β-chain variable region comprising the CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24-26, respectively, and (ii) an α-chain variable region comprising the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 42, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, or the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76, and 29, respectively.

[0196] In a preferred embodiment, the modified TCR comprises (i) an α-chain variable region comprising the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 42, 28, and 29, respectively, and (ii) a β-chain variable region comprising the CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24-26, respectively.

[0197] In a preferred embodiment, 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: 43, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0198] In a preferred embodiment, 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: 44, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0199] In a preferred embodiment, 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: 45, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0200] In a preferred embodiment, 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, 76, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0201] In a preferred embodiment, 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: 97, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0202] In a preferred embodiment, 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: 100, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0203] In a preferred embodiment, 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: 103, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0204] In a preferred embodiment, 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, 106, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0205] In a preferred embodiment, 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, 109, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0206] In a preferred embodiment, 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, 112, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0207] In a preferred embodiment, 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, 115, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0208] In a preferred embodiment, 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, 118, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0209] In a preferred embodiment, 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, 121, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0210] In a preferred embodiment, 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, 124, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0211] In a preferred embodiment, 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, 127, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24-26, respectively.

[0212] In some embodiments, the modified TCR (i) The variable region of the α chain, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 42, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 43, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 44, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 45, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 76, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 97, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 100, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 103, 28, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 106, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 109, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 112, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 115, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 118, and 29 respectively, CDR1α, CDR2α, and CDR3α with the amino acid sequences shown in SEQ ID NO: 27, 121, and 29 respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 140, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 146, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 165, 118, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 170, 76, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 118, and 29, respectively, or An α-chain variable region comprising CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, and (ii) a β-chain variable region, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 31, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 32, respectively, CDR1β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 33, respectively, CDR2β and CDR3β, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 88, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 25, and 94, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 25, and 131, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:24, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 153, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO:130, 91, and 26, respectively, The CDR1 of the amino acid sequences shown in SEQ ID NO: 130, 91, and 162, respectively β, CDR2β, and CDR3β, or The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 91, and 33, respectively, and the β-chain variable region

[0213] In some embodiments, the modified TCR is (i) an α-chain variable region comprising The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 42, 28, and 29, respectively The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 28, and 29, respectively The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 28, and 29, respectively The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, or The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76, and 29, respectively, and an α-chain variable region (ii) a β-chain variable region comprising The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 26, respectively The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 31, respectively The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 32, respectively The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 33, respectively The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 34, respectively The CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 35, respectively CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, or CDR1β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, CDR2β and CDR3β, and comprises a variable β-chain region.

[0214] In a preferred embodiment, the modified TCR comprises (i) a variable α-chain region comprising CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 76, and 29, respectively, and (ii) a variable β-chain region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 25, and 131, respectively.

[0215] In a preferred embodiment, 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: 44, 76, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 24, 91, and 35, respectively.

[0216] In a preferred embodiment, 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: 140, 141, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0217] In a preferred embodiment, 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: 146, 118, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0218] In a preferred embodiment, 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: 45, 149, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0219] In a preferred embodiment, 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: 152, 149, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 153, and 35, respectively.

[0220] In a preferred embodiment, 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: 43, 76, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 26, respectively.

[0221] In a preferred embodiment, 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: 45, 28, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 162, respectively.

[0222] In a preferred embodiment, 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: 165, 118, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0223] In a preferred embodiment, 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, 118, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0224] In a preferred embodiment, 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: 45, 141, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0225] In a preferred embodiment, 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: 170, 76, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0226] In a preferred embodiment, 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: 43, 118, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 130, 91, and 35, respectively.

[0227] In a preferred embodiment, 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: 152, 149, and 29, respectively, and (ii) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β having the amino acid sequences shown in SEQ ID NOs: 82, 91, and 33, respectively.

[0228] In some embodiments, the modified TCR has an α-chain variable region having the amino acid sequence shown 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 the amino acid sequences shown 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 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 any one of SEQ ID NOs: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, and includes a β-chain variable region having an amino acid sequence selected therefrom.

[0229] In some embodiments, the modified TCR is 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 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, 77, 98, 101, 104, 107, 110, 113, 116, 119, 122, 125, 128, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, and has an α-chain variable region having an amino acid sequence selected therefrom; and / or SEQ It includes a β-chain variable region having the amino acid sequence shown in 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.

[0230] In some embodiments, the modified TCR has an α-chain variable region having the amino acid sequence shown in 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 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, 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 SEQ It includes an amino acid sequence shown 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 selected from amino acid sequences 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. It includes a β-chain variable region having the amino acid sequence shown 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 selected from amino acid sequences 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.

[0231] As used herein, the term "sequence identity" means the degree to which two (nucleotide or amino acid) sequences have the same residue at the same position in an alignment, and is generally expressed as a percentage. Preferably, the identity is determined over the entire length of the aligned sequences. Thus, two copies having completely identical sequences have 100% identity, while sequences that are highly conserved and have deletions, additions or substitutions may have a lower degree of identity. One skilled in the art can 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. Thus, for example, the amino acid sequences of SEQ ID NO: 46 or 47 can be used as the "host sequence" or "reference sequence", while a different TCRα chain or β chain variable region amino acid sequence can be used as the "query sequence".

[0232] In some embodiments, the modified TCR comprises a variable region of the α chain having the amino acid sequence shown in SEQ ID NO: 46 and a variable region of the β chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NO: 48-64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175.

[0233] In some embodiments, the modified TCR is SEQ ID NO: 65- It comprises an α-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of 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 the amino acid sequence shown in SEQ ID NO: 47.

[0234] In some embodiments, the modified TCR comprises an α-chain variable region having an amino acid sequence selected from the amino acid sequences shown in 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 selected from the amino acid sequences shown in any one of SEQ ID NO: 48 - 64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175.

[0235] 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.

[0236] 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.

[0237] 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.

[0238] 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.

[0239] 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.

[0240] 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.

[0241] 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.

[0242] 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.

[0243] 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.

[0244] 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.

[0245] 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.

[0246] 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.

[0247] 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.

[0248] 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.

[0249] 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.

[0250] 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.

[0251] 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.

[0252] 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.

[0253] 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.

[0254] 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.

[0255] 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.

[0256] 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 having the amino acid sequence.

[0257] 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.

[0258] 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.

[0259] 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.

[0260] 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.

[0261] 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.

[0262] 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.

[0263] 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.

[0264] 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.

[0265] 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.

[0266] 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.

[0267] 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.

[0268] 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.

[0269] 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.

[0270] 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.

[0271] 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.

[0272] 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.

[0273] 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.

[0274] 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.

[0275] 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.

[0276] 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.

[0277] 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.

[0278] 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.

[0279] 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.

[0280] 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.

[0281] 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. sequence.

[0282] 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.

[0283] 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.

[0284] 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.

[0285] 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.

[0286] In a preferred embodiment, the modified TCR comprises an α-chain variable region having the amino acid sequence shown in SEQ ID NO: 173 and a β-chain variable region having the amino acid sequence shown in SEQ ID NO: 143.

[0287] 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.

[0288] In some embodiments, the modified TCR further comprises a TCR constant region.

[0289] As used herein, the term "constant region" may be a human constant region or may be from another species as in the case of obtaining a "chimeric" TCR. For example, the human α-chain and / or β-chain can be replaced ( "humanized") by their mouse counterparts. The mouse counterparts have been found to enhance surface expression of the human TCR and enhance more stable binding to the CD3 coreceptor by supporting preferential pairing of the TCR α-chain and β-chain.

[0290] In some embodiments, the TCR constant region is a mouse constant region or a human constant region.

[0291] It has been reported that adding a disulfide bond to the constant region promotes accurate pairing of the TCR α-chain and β-chain (Kuball J et al. Blood. 2007 Mar 15; 109(6):2331-8). Therefore, adding one or more cysteine modifications to the constant region to form a disulfide bond between the TCR α-chain and TCR β-chain is also contemplated herein.

[0292] In some embodiments, 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 mutation relative to the wild-type sequence to form a disulfide bond between the TCR α-chain and the TCR β-chain.

[0293] In some embodiments, the cysteine mutation is at one or more positions among position 48 in the constant region of the wild-type human TCRα chain, position 48 in the constant region of the wild-type mouse TCRα chain, position 57 in the constant region of the wild-type human TCRβ chain, and position 57 in the constant region of the wild-type mouse TCRβ chain.

[0294] The sequences of the wild-type TCR constant regions can be found in the publicly available database of the International Immunogenetics Information System (IMGT), such as the constant domain sequence of the α-chain of the TCR molecule being "TRAC*01" and the constant domain sequence of the β-chain of the TCR molecule being "TRBC1*01" or "TRBC2*01".

[0295] In the present invention, the positions of the amino acid sequences in the TCR constant region are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT). For example, if a certain amino acid in the constant region of the TCRα chain (TRAC) has a position number of 48 listed in IMGT, it is described herein as the 48th amino acid in the constant region of the TCRα chain (TRAC); if a certain amino acid in the constant region of the TCRβ chain (TRBC) has a position number of 57 listed in IMGT, it is described herein as the 57th amino acid in the constant region of the TCRβ chain (TRBC); and so on for the rest. In the present invention, when the sequence position numbers of other amino acids are specifically described, they will be explained accordingly.

[0296] In some embodiments, the constant region of the TCRα chain 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 TCRα 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 L L VI VL RIL. When the TCRα chain comprises a mouse constant region, the mouse constant region may comprise an LIV mutation such that the constant region (and / or transmembrane region) comprises the amino acid sequence L L V IV LRIL.

[0297] In some embodiments, the TCR α-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 β-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.

[0298] 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.

[0299] In some embodiments, the modified TCR may comprise a signal peptide sequence, for example, it may comprise a signal peptide sequence at the N-terminus of the α-chain and / or 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 shown in SEQ ID NO:74 (MKTFAGFSFLFLWLQLDCMS). The β-chain signal peptide sequence of the TCR of the present invention may have the amino acid sequence shown in SEQ ID NO:75 (MLLLLLLLGPGSGL).

[0300] In some embodiments, the modified TCR is as set forth in SEQ ID NO:69 A TCRα chain having an amino acid sequence shown, or having an amino acid sequence with 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 an amino acid sequence 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 selected from 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 TCRα chain having the amino acid sequence shown in SEQ ID NO:69 and a TCRβ chain having the amino acid sequence shown in any one selected from SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176. The amino acid sequences of SEQ ID NO:69 and 3-19 are shown below: JPEG2025518203000001.jpg190132JPEG2025518203000002.jpg195131JPEG2025518203000003.jpg120133

[0301] In some embodiments, the modified TCR comprises a TCR α chain having an amino acid sequence selected from the amino acid sequences shown in 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 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, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174; and / or a TCR β chain having the amino acid sequence shown 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. In some embodiments, the modified TCR comprises a TCR α chain having an amino acid sequence selected from the amino acid sequences shown in 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 β chain having the amino acid sequence shown in SEQ ID NO: 70.

[0302] The amino acid sequences of SEQ ID NOs: 20-23, 78 and 70 are shown below: MKTFAGFSFLFLWLQLDCMSRGEDVEQSLFLSVREGDSSVINCTYTDS LSTYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAGSGGGTDKLIFGTGTRLQVFPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO :20) MKTFAGFSFLFLWLQLDCMSRGEDVEQSLFLSVREGDSSVINCTYTDSS M TYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAGSGGGTDKLIFGTGTRLQVFPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO:21) MKTFAGFSFLFLWLQLDCMSRGEDVEQSLFLSVREGDSSVINCTYTDSS L TYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAGSGGGTDKLIFGTGTRLQVFPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO:22) MKTFAGFSFLFLWLQLDCMSRGEDVEQSLFLSVREGDSSVINCTYTDSS QTYLYWYKQEPGAGLQLLTYIFSNMDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAGSGGGTDKLIFGTGTRLQVFPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO:23) MKTFAGFSFLFLWLQLDCMSRGEDVEQSLFLSVREGDSSVINCTYTDSSSTYLYWYKQEPGAGLQLLTYIFS F MDMKQDQRLTVLLNKKDKHLSLRIADTQTGDSAIYFCAGSGGGTDKLIFGTGTRLQVFPNIQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDKCVLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO:78) MLLLLLLLGPGSGLGAVVSQHPSWVICKSGTSVKIECRSLDFQATTMFWYRQFPKQSLMLMATSNEGSKATYEQGVEKDKFLINHASLTLSTLTVTSAHPEDSSFYICSAREPGQGPYEQYFGPGTRLTVTEDLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGVCTDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNSSR (SEQ ID NO:70)

[0303] In some embodiments, the modified TCR has an amino acid sequence shown in 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 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, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174; and / or an amino acid sequence 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. In some embodiments, the modified TCR comprises a TCR α chain having an amino acid sequence selected from the amino acid sequences shown in 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 β chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 3-19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176.

[0304] In some embodiments, the modified TCR binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO:2), or a complex of said epitope and an MHC molecule.

[0305] The term "epitope" typically refers to a site on an antigen, and is usually a (poly)peptide recognized by a binding domain. The term "binding domain" most broadly refers to an "antigen-binding site", i.e., a domain that characterizes a molecule that binds / interacts with a specific epitope on an antigen target. An antigen target may contain a single epitope, but typically may contain at least two epitopes, and may contain any number of epitopes depending on the size, conformation, and type of the antigen. The term "epitope" typically includes linear epitopes and conformational epitopes. A linear epitope is a continuous epitope contained within the primary amino acid sequence, and usually contains at least two or a plurality of amino acids. A conformational epitope is formed by discontinuous amino acids juxtaposed by the folding of the target antigen, particularly the target (poly)peptide.

[0306] The TCR of the present invention can advantageously (specifically) recognize the MAGE-A1 antigen, particularly the epitope KVLEYVIKV (SEQ ID NO:2) on MAGE-A1 or a complex of said epitope and an MHC molecule, such as a complex of said epitope and HLA-A*02.

[0307] In some embodiments, the MHC molecule is of the HLA-A*02 type, such as HLA-A*02:01 type, HLA-A*02:03 type, HLA-A*02:05 type, HLA-A*02:06 type, HLA-A*02:07 type, HLA-A*02:10 type or HLA-A*02:11 type. In a preferred embodiment, the MHC molecule is of the HLA-A*02:01 type or HLA-A*02:05 type.

[0308] In some embodiments, the modified TCR is soluble or membrane-bound.

[0309] The TCR of the present invention can be provided in the form of a soluble TCR (sTCR). The soluble TCR is useful as a diagnostic tool and as a vector or "adapter" for targeting therapeutic agents or effector cells specifically to cancer cells expressing an antigen target recognized by the soluble TCR, for example. The soluble TCR is typically a fragment or construct comprising the α-chain and / or β-chain of the TCR or the variable region or CDR thereof, optionally stabilized by disulfide bonds or covalently linked by a suitable linker. Typically, the soluble TCR does not include, for example, a transmembrane region.

[0310] The TCR of the present invention can also be provided in a membrane-bound form. Typically, the membrane-bound TCR includes a transmembrane region for anchoring it on the cell membrane.

[0311] In some embodiments, the modified TCR is isolated or purified.

[0312] As used herein, the term "isolated or purified" means that the "isolated or purified" TCR has been identified, separated and / or recovered from the components of its manufacturing environment such that it does not contain, or substantially does not contain, other contaminant components from that manufacturing environment that could interfere with its therapeutic or diagnostic use. Contaminating components can include enzymes, hormones, and other protein or non-protein solutes. Thus, an "isolated or purified" TCR can be produced by at least one purification step that removes or substantially removes these contaminating components.

[0313] In another form, the present disclosure provides a nucleic acid encoding the modified TCR of the present invention.

[0314] In yet another form, the present disclosure provides a vector comprising the nucleic acid of the present invention.

[0315] As used herein, the term "vector" refers to, for example, a nucleic acid molecule used as a vehicle for introducing (exogenous) genetic material into a host cell, and the nucleic acid molecule that functions as a vector in the host cell can, for example, replicate and / or be expressed. The term "vector" encompasses, but is not limited to, plasmids, viral vectors (including retroviral vectors, lentiviral vectors, adenoviral vectors, vaccinia virus vectors, polyomavirus vectors, and adeno-associated virus (AAV) vectors), bacteriophages, phagemids, cosmids, and artificial chromosomes (including BACs and YACs). A vector itself is typically a nucleotide sequence, usually a DNA sequence that includes an insert (transgene) and a larger sequence as the vector "backbone". Engineered vectors typically include an origin of replication for autonomous replication in a host cell (optionally, stable expression of a polynucleotide), a selectable marker, and restriction enzyme cleavage sites (e.g., a polylinker site, MCS). A vector may further include a promoter, a genetic marker, a reporter gene, a target sequence, and / or a protein purification tag. As is known to those skilled in the art, a number of suitable vectors are known to those skilled in the art and 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 hereby incorporated by reference in its entirety.

[0316] In some embodiments, the vector is selected from a lentiviral vector, a retroviral vector, a plasmid, a DNA vector, an mRNA vector, a transposon-based vector, and an artificial chromosome.

[0317] In another form, the present disclosure provides a cell comprising a modified TCR, nucleic acid, or vector of the present invention.

[0318] As used herein, the term "cell" refers to any type of cell capable of expressing the modified TCR of the present invention. The cell may be a eukaryotic cell, such as a plant (which has no potential to grow into a plant), an animal, a fungus or an alga, or a prokaryotic cell, such as a bacterium or a protozoan. The cell may be a cultured cell or a primary cell, i.e., it may be directly isolated from an organism, e.g., a human. The cell may be an adherent cell or a floating cell, i.e., a cell that grows in suspension. Suitable host cells are known in the art and include, for example, DH5α E. coli cells, Chinese hamster ovary cells, monkey VERO cells, COS cells, HEK293 cells, etc. For the purpose of generating the TCR of the present invention, the cell is preferably a mammalian cell. Most preferably, the host cell is a human cell.

[0319] In some embodiments, the cell is selected from lymphocytes (e.g., T cells, NK cells), monocytes (e.g., PBMC) and stem cells.

[0320] As used herein, the term "stem cell" is a stem cell used for expressing the TCR of the present invention. For example, the stem cell can be a lymphocyte progenitor cell, an induced pluripotent stem cell (iPSC), or a hematopoietic stem cell (HSC). In some embodiments, the stem cell does not include embryonic stem cells obtained by destroying a human embryo and / or does not include totipotent stem cells used for generating and forming an animal individual. The introduction of a gene into a stem cell usually does not result in the expression of TCR on the cell surface because the surface of the stem cell does not express CD3 molecules. However, when the stem cell differentiates into a lymphoid precursor and migrates to the thymus, the expression of CD3 molecules starts, and the introduced TCR molecules are expressed on the surface of thymocytes.

[0321] In some embodiments, the stem cell is a lymphocyte progenitor cell or an induced pluripotent stem cell (iPSC).

[0322] In some embodiments, the cell is a T cell. The T cell can be any T cell, such as a cultured T cell, for example, a T cell derived from a primary T cell or a cultured T cell line, such as Jurkat, SupTl, etc., or a T cell obtained from a mammal. When obtained from a mammal, T cells can be obtained from many sources including, but not limited to, blood, bone marrow, lymph nodes, thymus, or other tissues and fluids. The T cells may be concentrated or purified. Preferably, the T cell is a human T cell. More preferably, the T cell is a T cell isolated from a human. The T cell can be any type of T cell, 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., and can be T cells at any stage of development. In some embodiments, the T cell does not express an endogenous TCR. The effector cells expressing the TCR of the present invention are expected to bind with high affinity to their antigen targets (preferably antigen epitopes presented by antigen presenting cells on HLA-A*02). The term "affinity" 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 thought to correlate with the in vivo effector ability of the cell expressing the TCR. By definition, a TCR-expressing cell with high affinity responds to very low antigen doses in in vitro tests, while such a cell with low binding affinity requires a larger amount of antigen before reaching an immune response similar to that of a high-affinity TCR-expressing cell. Thus, binding affinity can be considered a quantitative determinant of the activation threshold of cells expressing the TCR. This is determined by exposing such cells to different amounts of homologous antigen in vitro. Cells with a TCR having high binding affinity respond to low antigen doses. For example, when a TCR-expressing cell is co-cultured with a target antigen peptide-HLA-*02-expressing target cell, the EC50 for activating the target cell is less than 10 -5 M (e.g., 10 -5Less than M, for example 10 -6 M, 10 -7 M, 10 -8 M, or 10 -9 When (M) or (10), TCR-expressing cells are generally considered to bind to their antigenic targets with "high" binding affinity.

[0323] In yet another form, the present disclosure provides a method for producing a cell of the present invention, comprising the step of transducing or transfecting a cell with a vector of the present invention.

[0324] As used herein, the term "transfection" is a process of intentionally introducing a nucleic acid molecule or polynucleotide (including a vector) into a target cell. One example is RNA transfection, i.e., the process of introducing RNA (e.g., in vitro transcribed RNA, ivtRNA) into a host cell. The term is mainly used for non-viral methods in eukaryotic cells. The term "transduction" is generally used to describe the virus-mediated introduction of a nucleic acid molecule or polynucleotide. Transfection of animal cells usually involves creating transient pores or "holes" in the cell membrane to allow the uptake of substances. Transfection can be carried out using calcium phosphate, by electroporation, by cell extrusion, or by mixing a cationic lipid with a material to form 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.

[0325] In some embodiments, the method further comprises the step of growing and / or activating the cell before or after the transduction or transfection.

[0326] In another form, the present disclosure provides a conjugate comprising a modified TCR according to the present invention and an active agent bound or conjugated to said TCR.

[0327] 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 cytokines (e.g., IL-2 and IFN-γ), chemokines (e.g., IL-8), platelet factors (e.g., platelet factor 4), and complement promoters. Preferably, the therapeutic agent is selected from immunomodulators, radioactive compounds, enzymes, chemotherapeutic agents, and toxins. Other suitable therapeutic agents include small molecule cytotoxic agents, i.e., compounds having the ability to kill mammalian cells with a molecular weight of less than 700 daltons. Such compounds can also contain toxic metals that can have a cytotoxic effect. Further, it is understood that these small molecule cytotoxic agents also include prodrugs, i.e., compounds that disintegrate or are converted to release a cytotoxic agent under physiological conditions. Examples of such agents include cisplatin, maytansine derivatives, rachelmycin, calicheamicin, docetaxel, etoposide, gemcitabine, ifosfamide, irinotecan, melphalan, mitoxantrone, sophonem sodium photosensitizing factor II, temozolomide, topotecan, trimetreate glucuronate, crossostatin E, vincristine and doxorubicin; peptide cytotoxins, i.e., proteins or fragments thereof having the ability to kill mammalian cells; e.g., ricin, diphtheria toxin, Pseudomonas exotoxin A, Dnase and RNase; radionuclides, i.e., unstable isotopes of elements that emit one or more α or β particles or γ-rays during decay, e.g., iodine-131, rhenium-186, indium-111, yttrium-90, bismuth-210 and -213, actinium-225 and astatine-213; chelating agents useful for promoting binding to molecules or multimers thereof of these radionuclides; or heterologous protein domains, homologous protein domains, virus / bacterial protein domains, virus / bacterial peptides.

[0328] In yet another aspect, the present disclosure provides a modified TCR or antigen-binding fragment thereof of the invention. and one or more additional antigen-binding regions that bind to 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.

[0329] In some embodiments, each of the one or more additional antigens is independently selected from a tumor-associated antigen (TAA), a tumor-specific antigen (TSA), a viral antigen, an autoantigen, and an immune cell surface molecule.

[0330] Examples of tumor-associated antigens (TAA) include melanoma-associated antigens (e.g., gp100, MAGEA1, MAGEA3, MAGEA6, MAGEA4, MAGEA2, MAGEA12, MAGEA2B, MAGEA9B, MAGEA10, MAGEA11, MAGEB2, MAGEC1, MAGEC2), IGF2BP1, GNGT1, PI4K2B, CCR, NPSR1, COX7B2, ONE These include, but are not limited to, CUT3, 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.

[0331] Examples of tumor specific antigens (TSAs) include, but are not limited to, KRAS, TP53, PIK3CA, CTNNB1, EGFR, BRAF, and GNAS.

[0332] 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.

[0333] As used herein, the term "self-antigen" refers to an antigenic substance that causes changes in the structure and components of self-tissue cells induced by biological, physical, and chemical factors, thereby triggering an immune response against these self-tissue cell components. The release of sequestered antigens can give rise to an immune response. Certain tissue components within the human body, such as sperm, intraocular contents, and the brain, are not normally in contact with immune cells. Due to causes such as surgery, trauma, and infection, the barrier is disrupted, coming into contact with the immune system and resulting in an immune response such as autoimmune cross-linking ophthalmia. Changes in self-components can give rise to self-antigens. When self-components are subjected to the action of physical factors (e.g., cold, heat, ionizing radiation), chemical factors (e.g., drugs), or biological factors (e.g., bacteria, viruses, etc.), their antigenicity changes. The modified self-components stimulate the immune system to trigger an autoimmune response. For example, denatured IgG can stimulate the production of anti-denatured IgG antibodies (rheumatoid factor) and may cause rheumatoid arthritis. Clinically, the use of certain drugs can change the surface antigenicity of blood cells, potentially causing autoimmune hemolytic anemia or granulocytopenia. Cross-reaction due to common antigens can give rise to an immune response. Some bacteria and viruses have antigenic determinants similar to those of normal human tissue cells, and the autoantibodies and sensitized lymphocytes produced against these antigenic determinants of bacteria and viruses can cross-react with self-tissue cells, potentially causing autoimmune diseases. For example, some antigen proteins of group A hemolytic streptococci have common antigens with human endocardium and glomerular basement membrane. The immune response induced by coxsackievirus infection can attack pancreatic beta cells and cause diabetes.

[0334] In some embodiments, the self-antigen is erythematosus, diabetes caused by autoimmune pancreatic islets, ankylosing spondylitis, rheumatoid arthritis.

[0335] Examples of immune cell surface molecules include 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, CD117, 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, but are not limited thereto. 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 anti-CD3 TCR, preferably from the antigen-binding region of an anti-CD3 antibody. In a preferred embodiment, the present disclosure provides a bispecific molecule comprising the modified TCR of the present invention or an antigen-binding fragment thereof and the antigen-binding region of an anti-CD3 antibody.

[0336] In another form, 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.

[0337] The term "composition" particularly refers to a composition suitable for administration to humans. However, the term usually also 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., they can induce the desired therapeutic effect in the recipient without causing undesirable local or systemic effects. The pharmaceutically acceptable compositions of the present invention can be, for example, sterile. Specifically, the term "pharmaceutically acceptable" can mean being approved by a regulatory authority or other generally recognized pharmacopoeia for use in animals, more specifically in humans.

[0338] The term "excipient" includes fillers, binders, disintegrants, coating agents, adsorbents, anti-adhesion agents, flow promoters, preservatives, antioxidants, flavoring agents, coloring agents, sweetening agents, solvents, co-solvents, buffers, chelating agents, viscosity-imparting agents, surfactants, diluents, wetting agents, carriers, diluents, preservatives, emulsifiers, stabilizers, and isotonic agents. Those skilled in the art know how to select excipients suitable for manufacturing the compositions of the present invention. Exemplary carriers for use in the compositions of the present invention include physiological saline, buffered physiological saline, dextrose, and water. Generally, the selection of appropriate excipients depends particularly on the active agent used, the disease being treated, and the desired dosage form of the composition. Depending on the active agent used (e.g., soluble TCR), the compositions of the present disclosure can be in various forms, such as solid, liquid, gas, or lyophilized forms, particularly in the form of ointments, creams, transdermal patches, gels, powders, tablets, solutions, aerosols, particles, pills, suspensions, emulsions, capsules, syrups, liquids, elixirs, extracts, tinctures, or fluid extracts, or in a form particularly suitable for the desired method of administration. The manufacturing processes of pharmaceuticals known in the present invention are shown in the 22nd edition of Remington’s Pharmaceutical Sciences (Ed. Maack Publishing Co, Easton, Pa., 2012) and can include, for example, conventional mixing, dissolving, granulating, sugar coating, pulverizing, emulsifying, encapsulating, embedding, or lyophilizing processes. For example, the compositions containing host cells or soluble TCR described herein are usually provided in liquid form and preferably contain a pharmaceutically acceptable buffer.

[0339] In some embodiments, the composition further comprises a second therapeutic agent, and preferably, the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs.

[0340] Preferred examples of the second therapeutic agent include known anti-cancer agents such as cisplatin, maytansine derivatives, rachelmycin, calicheamicin, docetaxel, etoposide, gemcitabine, ifosfamide, irinotecan, melphalan, mitoxantrone, porfimer sodium Photofrin II (sorfimer sodium photofrin II), temozolomide, topotecan, trimetrexate glucuronate, auristatin E, vincristine and doxorubicin; and peptide cytotoxins such as ricin, diphtheria toxin, Pseudomonas exotoxin A, DNA enzymes and RNA enzymes; radionuclides such as iodine 131, rhenium 186, indium 111, yttrium 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; virus / bacterial protein domains and virus / bacterial peptides. In another form, the present disclosure provides a method of treating MAGE-A1 positive cancer in a subject, the method comprising administering to the subject an effective amount of the modified TCR of the present invention.

[0341] In yet another form, the present disclosure provides a method of treating MAGE-A1 positive cancer in a subject, the method comprising administering to the subject an effective amount of the cells of the present invention.

[0342] As used herein, the term "treatment" includes therapeutic or prophylactic treatment in a subject in need thereof. "Therapeutic or prophylactic treatment" includes prophylactic treatment aimed at completely preventing clinical and / or pathological symptoms, or therapeutic treatment aimed at improving or alleviating clinical and / or pathological symptoms. Thus, the term "treatment" also includes improvement or prevention of a disease.

[0343] 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" can vary depending on the compound, the disease and its severity, as well as the age, weight, etc. of the subject being treated. A "therapeutically effective amount" refers to an effective amount for therapeutic treatment. A "prophylactically effective amount" refers to an effective amount for prophylactic treatment.

[0344] Therapeutic efficacy and toxicity can be determined by standard procedures in cell cultures or experimental animals, for example, ED50 (the therapeutically effective dose in 50% of the population) and LD50 (the lethal dose for 50% of the population). The dose ratio between the therapeutic and toxic effects is the therapeutic index, which can be expressed as the ratio of ED50 / LD50. Pharmaceutical compositions showing a large therapeutic index are preferred.

[0345] One of ordinary skill in the art can determine the exact dose of the TCR or cell to be administered using known techniques. The appropriate dose 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 prophylactic response in the subject or animal within a reasonable time frame. For example, the dose of the TCR of the present invention should be sufficient to bind to a cancer antigen or detect, treat or prevent cancer over a period of about 2 hours or more from the time of administration, for example, 12 hours to 24 hours or more (e.g., 1 month, 2 months, 3 months, 6 months, 12 months, 24 months, etc.). In certain embodiments, the period may be even longer. As is known in the art, treatment objectives (e.g., reduction of acute exacerbation of a disease), route of administration, time and frequency, time and frequency of the dosage formulation, age, weight, general health status, gender, diet, severity of the disease state, drug combination, response sensitivity, and adjustment of tolerance / response to treatment may be required.

[0346] Many assays for determining dosage are known in the art. For the purposes of the present invention Thus, an assay can be used to determine the starting dose to be administered to a mammal, including comparing the degree to which target cells are lysed or the degree of IFN-γ secreted by such T cells after administering a given dose of T cells expressing the TCR of the present invention to a group of mammals (administering different doses of T cells). After administration of a dose, the degree to which target cells are lysed or the degree to which IFN-γ is secreted can be determined by methods known in the art. The dose 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 be associated with administration of the TCR or cells of the present invention. Generally, the attending physician will consider various factors such as age, weight, general health, diet, gender, the active agent being administered, the route of administration, and the severity of the treatment condition to determine the dose of the TCR or cells of the present invention to be administered to an individual patient. In some embodiments of the treatment methods of the present disclosure, the number of cells administered per injection can vary, for example, from about 1×10 6 to about 1×10 12 cells or more. In some embodiments, fewer than 1x10 6 cells can be administered.

[0347] It should be recognized that treatment may require a single or multiple administrations of a therapeutically effective dose of the active agent of the present invention. For example, depending on the formulation, half-life, and clearance of a particular composition, some compositions can be administered once every 3 to 4 days, weekly, or every 2 weeks, or within 1 month. The compositions of the present disclosure are suitable for administration by various routes. Typically, administration is achieved parenterally. Parenteral delivery methods include topical, intraarterial, intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal, intrauterine, intravaginal, sublingual, or intranasal administration.

[0348] The terms "subject" or "individual" or "animal" or "patient" are used interchangeably herein and 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, the TCRs, nucleic acids, vectors, host cells, and pharmaceutical compositions provided herein are particularly contemplated for use in the treatment of human subjects, particularly HLA-A*02 positive subjects, 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 positive subjects, preferably HLA-A*02:01 or HLA-A*02:05 positive subjects.

[0349] In some embodiments of the disclosed methods of treatment, the cells are autologous or allogeneic to the subject.

[0350] In some embodiments, the method comprises: (i) isolating a cell-containing sample from the subject; (ii) transducing or transfecting the cells with a vector of the invention; and (iii) administering the cells obtained in step (ii) to the subject. In some embodiments, the method further comprises knocking out the endogenous TCR in the cells after step (i) and before step (ii).

[0351] 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 carcinoma of the bladder), ovarian cancer (e.g., serous cystadenocarcinoma of the ovary), head and neck cancer (e.g., squamous cell carcinoma of the head and neck), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma. In some embodiments, the method further comprises administering a second therapeutic agent, preferably, the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs. Preferred examples of the second therapeutic agent are as described above.

[0352] In some embodiments of the treatment methods of the present disclosure, 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.

[0353] In another form, the present disclosure provides a method for detecting (e.g., diagnosing) MAGE-A1 positive cancer in a subject, the method comprising: (i) contacting a sample obtained from the subject with the TCR, cell or conjugate of the present invention; and (ii) detecting the presence of 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., bladder urothelial carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma.

[0354] 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 a 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 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, the detectable label is an enzyme or a catalytically active fragment thereof, a radionuclide, a fluorescent, phosphorescent or chemiluminescent molecule.

[0355] In yet another aspect, the present disclosure provides a kit comprising a TCR or conjugate of the present invention for detecting the presence of a positive epitope in a test sample, wherein the epitope comprises an epitope comprising KVLEYVIKV (SEQ ID NO:2).

[0356] In some embodiments, the kit is a kit for detecting (e.g., diagnosing) MAGE-A1 positive cancer in a subject and comprises a TCR or conjugate of the present invention. 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., serous cystadenocarcinoma of the ovary), head and neck cancer (e.g., squamous cell carcinoma of the head and neck), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma. In some embodiments, the conjugate comprises a 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. In some embodiments, the kit can further comprise instructions regarding how to use the kit.

[0357] In another aspect, the present disclosure provides the use of the modified TCR, nucleic acid, vector, cell, conjugate, bispecific antigen-binding molecule or multispecific antigen-binding molecule, or composition of the present invention in the manufacture of a medicament for the treatment of MAGE-A1 positive cancer in a subject.

[0358] In yet another aspect, the present disclosure provides the modified TCR, nucleic acid, vector, cell, conjugate, bispecific antigen-binding molecule or multispecific antigen-binding molecule, or composition of the present invention for use in the treatment of MAGE-A1 positive cancer in a subject.

[0359] In another aspect, the present disclosure provides the use of the modified TCR or conjugate of the present invention in the manufacture of 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).

[0360] In some embodiments, the present disclosure provides the use of the modified TCR or conjugate of the present invention in the manufacture of a kit for detecting (e.g., diagnosing) MAGE-A1 positive cancer in a subject.

[0361] In yet another aspect, the present disclosure provides a modified TCR or conjugate of the 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), and for example, detecting (e.g., diagnosing) MAGE-A1 positive cancer in a subject.

[0362] In some embodiments of the use of the present disclosure, 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., bladder urothelial carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymoma.

[0363] In some embodiments, the subject has an HLA-A*02:01 allele, for example, 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.

[0364] Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. Note that this example is for explaining the present invention and does not limit the scope of the present invention. In the following examples, experimental methods for which specific conditions are not specified are carried out according to the conditions described in, for example, Sambrook and Russel et al., Molecular Cloning: A Laboratory Manual (3rd Edition) (2001), Cold Spring Harbor Laboratory Press, or the conditions recommended by the manufacturer, etc., which are ordinary conditions in the art. The test materials and reagents used in the following examples are commercially available unless otherwise specified.

[0365] Example 1. Screening for Substituted TCRs with Improved Affinity The inventors replaced the complementarity-determining region (CDR) region of ht27 using an in vitro substitution method with the human-derived TCR ht27 sequence as a template, and obtained high-affinity substitution clones by in vitro positive screening of the substituted TCR. By this method, the preferred TCR sequences screened contained amino acid substitutions at Gln29, Ala36, Thr37, Thr38 at the position of CDR1β of ht27β chain, Ser56, Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65 at the position of CDR2β, and Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114 and Glu115 at the position of CDR3β. The specific amino acid substitutions were Q29P, A36I, T37E, T38R, S56T, N57D, N57E, N57H, N57K, N57Q, N57R, N57S, N57T, E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, E58W, G59D, G59E, G59 H, G59K, G59Q, G59R, G59S, G59T, G59 P 、G59 L, 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, E108T, E108I, E108K, E108P, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, Q111H, G112E, P113H, substitution of EPGQGP at positions 108 - 113 with VNDGT, Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, Y114S, E115I. The screened preferred TCR sequences contain amino acid substitutions at positions Asp27, Ser28, Ser29, Ser36, Thr37, Tyr38 of CDR1α and positions Ile56, Phe57, Ser58, Asn59, Met63, Asp64 of CDR2α of the ht27 α-chain. Specific amino acid substitutions include S28Y, S28P, S29Y, S29T, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W, S29N, 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 screened preferred TCR sequences also include amino acid substitutions at position Thr26 of the α-chain framework region 1 (FR1α), such as T26A, T26V, etc.

[0366] Each of the three CDR regions of the TCR α-chain and β-chain binds to different sites of the MHC-polypeptide complex and acts on the MHC-polypeptide complex in a relatively independent form. Mutations in the above CDR regions, whether single mutations or combined mutations, are effective in improving the affinity of the TCR for the wild-type TCR. Depending on the combination of mutations, a synergistic effect that achieves a higher affinity can also be brought about.

[0367] Representative substituted TCRs are shown in Table 1 below. Table 1 shows the amino acid substitution regions and substitution types of the representative substituted TCRs of the present invention with respect to the wild-type ht27 TCR.

[0368] [Table 1] JPEG2025518203000005.jpg37133

[0369] Furthermore, screening was performed to obtain mutant TCRs having an affinity up to the pM level by combining mutations in different CDR regions as described above. The positions of amino acid substitutions in the screened TCR sequences include Asp27, Phe28, Gln29, Ala36, Thr37, Thr38 at the position of CDR1β of the ht27 β-chain, Ser56, Asn57, Glu58, Gly59, Ser63, Lys64 and Ala65 at the position of CDR2β, and Ser105, Ala106, Arg107, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114 and Glu115 at the position of CDR3β, and Asp27, Ser28, Ser29, Ser36, Thr37, Tyr38 at the position of CDR1α of the ht27 α-chain, and Ile56, Phe57, Ser58, Asn59, Met63, Asp64, Met65 at the position of CDR2α, and also includes amino acid substitutions at the position of FR1α Thr26. Representative substituted TCRs are shown in Table 2 below. Table 2 shows the amino acid substitution regions of the representative substituted TCRs of the present invention and the substitution types with respect to the wild-type ht27 TCR.

[0370]

Table 2

[0371] The CDR sequences of the wild-type ht27 TCR and the amino acid sequences of the variable regions of the α-chain and β-chain are shown in Table 3 below. The CDR sequences of the representative substituted TCRs of the present invention and the amino acid sequences of the variable regions of the α-chain and β-chain are shown in Table 4 below.

[0372]

Table 3

[0373]

Table 4

[0374] To increase the correct pairing and expression of the exogenous TCR on the surface of human-derived T cells, the variable regions of the α-chain and β-chain of the TCR are fused to the mouse-derived TCR constant region, and the classical α-chain C region "LIV" substitution (i.e., the constant region has the amino acid sequence L L V IVIntroduce amino acid substitutions into the mouse-derived TCR α-chain constant region so as to contain LRIL. (shown in bold), as well as classical α-chain T48C and β-chain S57C substitutions (shown in bold and underlined) were introduced.

[0375] Amino acid sequence of the α-chain constant region: IQNPEPAVYQLKDPRSQDSTLCLFTDFDSQINVPKTMESGTFITDK C VLDMKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLWSS (SEQ ID NO: 71)

[0376] Amino acid sequence of the β-chain constant region: DLRNVTPPKVSLFEPSKAEIANKQKATLVCLARGFFPDHVELSWWVNGKEVHSGV C TDPQAYKESNYSYCLSSRLRVSATFWHNPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLVVMAMVKRKNSSR (SEQ ID NO: 72)

[0377] The α-chain and β-chain of the TCR (the N-terminals of the α-chain and β-chain contain the signal peptide sequences MKTFAGFSFLFLWLQLDCMS (SEQ ID NO:74) and MLLLLLLLGPGSGL (SEQ ID NO:75), respectively) were tandemly constructed into a lentiviral expression vector via a 2A self-cleaving peptide (ATNFSLLKQAGDVEENPGP (SEQ ID NO:73)) in the order of β-T2A-α. ht27-WT Taking the 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 shown underlined, and the 2A self-cleaving peptide is shown in italics). JPEG2025518203000022.jpg53140

[0378] The natural human-derived TCR ht27WT and the substituted TCR sequences of the present invention were each cloned into a TCR tandem expression lentiviral vector, the lentivirus was packaged, reporter cells JKR9 were transduced, and then TCR expression-positive cells were selected. T2 cells positive for HLA-A*02:01 were loaded with MAGE-A1 target antigen peptides (KVLEYVIKV (SEQ ID NO:2)) at various concentrations and then activated by co-culturing with JKR9 reporter cells transduced with TCR ht27WT and the substituted TCR of the present invention, respectively. After 16 hours of activation, the stained reporter gene was analyzed by flow cytometry. The results of activation on the reporter cells of the high-affinity TCR of the present invention are shown in FIGS. 1 and 2. From the results in FIG. 2, the EC50 value was calculated, the EC50 value of ht27-WT was set to 1, and the results of calculating the EC50 enhancement multiple of the substituted TCR relative to ht27-WT are shown in Table 5.

[0379] [Table 5] JPEG2025518203000024.jpg27134

[0380] Among them, the mutant TCRs combined with mutations in different CDR regions have significantly better activation functions than the wild-type TCR ht27, have pM-level affinity, and their EC50 values are as shown in Table 6 below.

[0381] [Table 6]

[0382] The above results show that TCRs having amino acid substitutions at one or more of the amino acid positions Asn57, Glu58, Gly59, Ser63, Lys64, Ala65, Ser105, Gln111, Tyr114, Ser29, Ser36, Asn59 are wild-type TCRs It is shown that compared to ht27, it exhibits significantly improved affinity. These amino acid substitutions can further be combined to generate substituted TCRs with higher affinity.

[0383] However, after transducing combinatorial mutant TCRs into JKR9 reporter cells, a non-specific activation reaction occurred with T2 cells not carrying the target peptide (Figure 1), which is probably due to the excessive affinity of these TCRs. Similar non-specific activation was also observed by performing the above-mentioned flow analysis using prior art ultra-high affinity mutant TCRs targeting gp100 (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 the presence of non-specific activation, the Kimmtrak molecule (trade name: Tebentafusp-tebn), a soluble TCR protein drug manufactured using the mutant TCR gp100-b27a8, obtained marketing approval for the treatment of metastatic uveal melanoma. This indicates the pharmaceutical value of ultra-high affinity mutant TCRs (KD-SPR value < 1 nM).

[0384] Example 2. Evaluation of the Affinity of Ultra-High Affinity Mutant TCRs by Dextramer Staining Multimers (Dextramer) are multimers of pMHC molecules that can be used to find TCR sequences that specifically recognize a particular antigen peptide from a T cell library in in vitro assays. Specific binding between Dextramer and TCR enables flow detection and sorting by fluorescence labeling. On the other hand, since the binding strength between Dextramer and TCR is positively correlated with TCR affinity, mutant TCRs with extremely high affinity can be screened by the Dextramer staining method, and the affinity of mutant TCRs can be evaluated.

[0385] To verify the feasibility of this method, mutant TCRs (gp100-b03a8, gp100-b28a2, gp100-b28a8, gp100-b33a2, gp100-b17a2, gp100-b10a2, gp100 -b23a2, gp100-b26a2, gp100-b26a8, gp100-b27a2, gp100-b27a8) with known affinity for the target gp100 were used for the test. The affinity of each mutant of the known gp100-TCR measured by SPR is as follows.

[0386]

Table 7

[0387] The verification method is as follows. First, gp100 mutant TCRs with known affinity were transduced into JKR9 cells via lentivirus to construct mutant TCR-expressing cell lines. Next, Dextramer was produced using the gp100 target antigen peptide (YLEPGPVTV, SEQ ID NO: 177). The Dextramer was diluted to 10 times the concentration with PBS and diluted in a total of four concentration gradients (final staining concentration 2 nM to 0.002 nM). Then, JKR9 cells transduced with mutant TCRs having different affinities were stained in parallel with Dextramer at different concentrations. The staining positive rate and MFI of each experimental group were recorded by flow cytometry. The results are shown in Figure 4. Finally, by summarizing the data of different experimental groups, it was analyzed whether the Dextramer staining positive rate and the average staining intensity (MFI) have a correlation with the affinity measured by SPR, respectively.

[0388] As a result, it was shown that as the concentration of Dextramer used in staining decreased, the high-affinity TCR staining superiority gradually appeared in relation to the mutant TCR affinity, both in terms of the staining positive rate and MFI (Figure 4). After further analyzing the staining data, it was found that when using Dextramer with a concentration of less than 0.2 nM, the staining intensity and the affinity of TCR showed a linear correlation (Figure 5). When performing a correlation analysis of the EC50 of the MFI of the limiting low-concentration (0.02 nM) Dextramer staining and the MFI of the Dextramer gradient staining with respect to the true affinity (KD-SPR) of SPR measurement, it was found that the results of SPR measurement could react well using both the MFI of the limiting low-concentration Dextramer staining and the MFI of the Dextramer gradient staining, and a linear relationship formula between KD-SPR and Dextramer staining MFI (Figure 6) was obtained. These results indicate that Dextramer staining can be used to evaluate the affinity of mutant TCRs.

[0389] After confirming the affinity of the mutant TCR targeting MAGE-A1 of the present invention by the above method, it was evaluated using the above Dextramer staining method. The results are shown in Figures 7 to 8.

[0390] As a result, the mutant TCR of the present invention showed strong staining positivity under extremely low-concentration Dextramer staining conditions, and the staining intensity (MFI) increased with the increase in Dextramer concentration, indicating that the mutant TCR of the present invention has extremely high affinity for the target-pMHC complex.

[0391] As described above, using the linear relationship formula between KD-SPR and Dextramer staining MFI (Figure 6), the MFI of the Dextramer staining of the mutant TCR of the present invention was substituted into the formula, and the results of estimating the KD-SPR value of the mutant TCR are shown in the following table. The results indicate that the estimated SPR values of all mutant TCRs have reached the 10 pM level, indicating the very high affinity of the mutant TCR of the present invention.

[0392]

Table 8

[0393] Example 3: Evaluation of the cell membrane expression stability of TCR with optimized affinity Lentiviruses (MOI = 10) producing ht27WT and TCR with optimized affinity were packaged in 293T cells, transduced into PBMC, and then cultured for proliferation until collected on the 11th day. The cell proliferation at harvest was good, with a proliferation multiple of 70 - 150 times, and the cell viability rate was >90% for both. To confirm T cells while demonstrating the success of TCR transduction, cells were stained with human CD3-FITC antibody; to detect the expression rate of TCR-T, cells were stained with anti-mouse TCRβ-APC antibody; and to examine the CD4 / CD8 ratio, cells were stained with CD4-APC and CD8-PE-Cy7 antibodies.

[0394] The results of flow cytometry analysis showed that, under the same transduction and culture conditions, the CD4 / CD8 expression ratio of each cell after transduction with TCR with optimized affinity was close to 1:1, and the TCR positive rate was over 75% and between 75 - 90% (Figure 9). This indicates that the TCR molecule with optimized affinity has very good membrane expression stability, suggesting that it potentially has very good antigen reactivity and antitumor activity.

[0395] Example 4: Efficient specific killing of TCR-T cells with optimized affinity against antigen-positive tumor cells T cells expressing TCR or ht27WT with optimized affinity as effector cells were used, and PBMCs without transduced TCR were cultured in parallel and then used as negative controls for effector cells. Hela-A2-eGFP-LUC (MAGE-A1 positive, HLA-A*02:01 positive) was used as HLA-antigen peptide compatible positive target cells; Siha-eGFP-LUC cells (MAGE-A1 negative, HLA-A*02:01 negative) were used as HLA-antigen peptide incompatible 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 a luciferase substrate was added to detect viable target cells. The results of calculating the percentage of target cells killed from the remaining target cells are shown in Figure 10.

[0396] As a result, TCR-T cells with optimized affinity showed obvious killing activity against Hela-A2-eGFP-LUC positive target cells and had significantly higher killing activity than Ht27WT TCR-T cells; TCR-T cells with optimized affinity had no killing effect on negative target cells. These results indicate that TCR-T cells with optimized affinity have the ability to efficiently and specifically kill target cells compatible with HLA-antigen peptides, and their killing activity increases with the increase in affinity.

[0397] Example 5. Specific INF-γ secretion of TCR-T cells with optimized affinity for antigen-positive tumor cells T cells expressing TCR or ht27WT with optimized affinity as effector cells were used, and PBMCs without transduced TCR were cultured in parallel and then used as negative controls for effector cells. Hela-A2-eGFP-LUC cells and A375-eGFP-LUC cells (MAGE-A1 positive, HLA-A*02:01 positive) were used as HLA-antigen peptide compatible positive target cells; Siha-eGFP-LUC cells (MAG E-A1 negative and HLA-A*02:01 negative) were used as HLA antigen-peptide mismatched negative target cells. Target cells different from effector cells were incubated at an effector-target ratio (E:T) of 1:1 for 16 to 20 hours, and the results of detecting IFN-γ secretion in the supernatant using an IFN-γ ELISA kit (ExCell, Cat#EH008-96) are shown in Figure 11.

[0398] As a result, after co-culturing the TCR-T cells with optimized affinity with the positive target cells A375 or Hela-A2, obvious IFN-γ secretion was detected, showing a significantly higher IFN-γ secretion level than that of Ht27WT TCR-T cells, indicating that no IFN-γ secretion was detected in HLA-antigen peptide mismatched negative target cells. These results show that TCR-T cells with optimized affinity have a good specific killing effect on target cells with HLA-antigen peptide compatibility, and their specific killing effect also increases with the increase in affinity.

[0399] Example 6: Nonspecific activation of TCR-T cells with optimized affinity against T2-KO-TAP1 cell lines expressing different HLA typings To detect whether TCR-T cells with optimized affinity cause alloreactivity, that is, cross-reactivity with other mismatched HLA and antigen independence, a T2-KO-TAP1 cell line (knocking out HLA and expressing the antigen peptide transporter protein TAP1) was constructed, and 66 different HLAs were expressed by this cell line respectively, and the selected high-frequency HLAs A, B, and C could cover more than 90% of the human population. After transducing various HLA genes into T2-KO-TAP1 cells and sorting by flow cytometry, the HLA expression exceeded 95%. Using 1 × 10 5 T cells expressing TCR with optimized affinity as effector cells, 5 1 × 10 -7T2 cells (positive for HLA-A*02:01) carrying the MAGE-A1 polypeptide of M were used as a positive control for target cells, and the results are shown in Figure 12.

[0400] The results showed that no IFN-γ secretion was detected after co-culture with Ht27M1001 (388-fold increase in affinity), Ht27M1005 (11-fold increase in affinity), and all T2-KO-TAP1-HLA cells, indicating that Ht27M1001 and Ht27M1005 have good specificity.

[0401] Example 7: Non-specific activation of TCRs with optimized affinity for antigen-negative target cells from different tissues and T2-KO-TAP1 cell lines expressing different HLA typings To further determine the specificity of TCRs with optimized affinity for recognizing target antigens, non-specific activation of TCRs with optimized affinity for antigen-negative target cells from different tissues and T2-KO-TAP1 cell lines expressing different HLA typings was detected. The specific experimental procedures are as follows. Construction of an off-target reaction detection tool: Referring to the transcriptome sequencing data TPM (Transcripts Per Million mapped reads) of the Cancer Cell Line Encyclopedia (CCLE) database, a target gene TPM = 0 was defined as negative expression of the target gene. As antigen-negative target cells, nine cell lines with negative MAGE-A1 gene expression derived from different tissues were screened, and the HLA-A*02:01 gene was transduced into these cell lines as tool cells for detecting off-target reactions.

[0402] Construction of an allogeneic reaction detection tool: The endogenous HLA-B*51:01 and HLA-C*01:02 genes of T2 cells were knocked out to obtain endogenous HLA class I molecules (T2KO cells). Suppress the expression; then overexpress the TAP1 / 2 gene in T2 cells to construct a tool cell (T2KO-TAP) having a complete function of endogenous antigen presentation; finally, transduce different high-frequency HLAs into T2KO-TAP to construct T2-KO-TAP1 cell lines with different typings as tool cells for detecting allogeneic reactions. Specifically, it was constructed as described in Example 6.

[0403] Detection of non-specific reactivity: During the detection, JKR9 reporter cells transduced with TCR with optimized affinity were co-cultured with antigen-negative target cells overexpressing HLA-A*02:01 and T2-KO-TAP1 cell lines expressing different HLA typings, respectively. After culturing for 16 hours, whether the JKR9 reporter cells transduced with high-affinity mutant TCR could be activated by the above tool cells, and the presence of off-target reaction or allogeneic reaction of the mutant TCR was determined by flow cytometry. HLA-antigen peptide-compatible positive target cells A375-A1-A2 cells (MAGE-A1 positive, HLA-A*02:01 positive) were used as a positive control. The results of non-specific activation of the high-affinity TCR according to the present invention are shown in the following table.

[0404]

Table 9

[0405] The results showed that in none of ht27M1001, ht27M1002, ht27M1003, ht27M1004, ht27M1005, ht27M1006, ht27M1009, ht27M1010, ht27M1011, ht27M1012, ht27M1013, ht27M1018, ht27M1019, ht27M1128, ht27M1166, ht27M1169, ht271170, ht27M1226, ht27M1240, ht27M1243, ht27M1245, ht27M1285, ht27M1290, ht27M1369, ht27M1378, ht27M1414, ht27M1457, ht27M1557, ht27M1387, ht27M1398, ht27M1401, ht27M1374, ht27M1381, ht27M1360, ht27M1362, ht27M1363, ht27M1365, ht27M1418, ht27M1436 was the non-specific activation of HLA-compatible antigen-negative target cells and T2-KO-TAP1 cell lines expressing different HLA typings detected, indicating that these TCRs with optimized affinities have good specificity.

[0406] The present invention is described in various published patents, published patent applications, periodicals, etc., all of which are hereby incorporated by reference. In case of any conflict between any of the incorporated references and this specification, this specification shall prevail. Further, any particular embodiment of the present invention that falls within the scope of the prior art may be explicitly excluded from any one or more of the claims. Since the described embodiments are considered to be known to those skilled in the art, they may be excluded even if the described exclusions are not explicitly described in the present invention. Any particular embodiment of the present invention may be excluded from any claim for any reason, whether or not it is related to the existence of the prior art.

[0407] Although the present invention has been described with reference to its specific embodiments, those skilled in the art will understand that various changes can be made and equivalents can be substituted without departing from the true spirit and scope of the present invention. Furthermore, many modifications can be made to adapt a particular situation, material, composition, method, process to the purpose, spirit and scope of the present invention. All such modifications are intended to fall within the scope of the claims.

Claims

1. A modified T cell receptor (TCR) comprising a variable α-chain region containing CDR1α, CDR2α, and CDR3α, and a variable β-chain region containing CDR1β, CDR2β, and CDR3β, wherein, compared to the parental TCR, the following amino acid substitutions are made: (i) Amino acid substitutions in the variable β-chain region, wherein (a) Amino acid substitutions at one or more positions of Gln29, Ala36, Thr37, and Thr38 in CDR1β, (b) Amino acid substitutions at one or more positions of Ser56, Asn57, Glu58, Gly59, Ser63, Lys64, and Ala65 in CDR2β, and / or (c) Amino acid substitutions at one or more positions of Ser105, Glu108, Pro109, Gly110, Gln111, Gly112, Pro113, Tyr114, and Glu115 in CDR3β, and / or (ii) Amino acid substitutions in the variable α-chain region, wherein (d) Amino acid substitutions at one or more positions of Asp27, Ser28, Ser29, Ser36, Thr37, and Tyr38 in CDR1α, (e) Amino acid substitutions at the positions of Ile56, Phe57, Ser58, Asn59, Met63, and Asp64 in CDR2α, and / or (f) Amino acid substitution at the Thr26 position in FR1α, wherein the parental TCR comprises CDR1β with the amino acid sequence shown in SEQ ID NO: 24, CDR2β with the amino acid sequence shown in SEQ ID NO: 25, CDR3β with the amino acid sequence shown in SEQ ID NO: 26, CDR1α with the amino acid sequence shown in SEQ ID NO: 27, CDR2α with the amino acid sequence shown in SEQ ID NO: 28, and CDR3α with the amino acid sequence shown in SEQ ID NO: 29, and the amino acid positions are numbered according to the nomenclature rules of the International Immunogenetics Information System (IMGT).

2. The modified TCR has the following amino acid substitutions (1) to (33) compared to the parental TCR: (1) Amino acid substitution at the Gln29 position in CDR1β selected from Q29E and Q29P; (2) Amino acid substitution A36I at the Ala36 position in CDR1β; (3) Amino acid substitution T37E at the Thr37 position in CDR1β; (4) Amino acid substitution T38R at the Thr38 position in CDR1β; (5) Amino acid substitution S56T at the Ser56 position in CDR2β; (6) An amino acid substitution at the Asn57 position in CDR2β selected from N57D, N57E, N57H, N57K, N57Q, N57R, N57S, and N57T; (7) An amino acid substitution at the Glu58 position in CDR2β selected from E58A, E58F, E58I, E58L, E58M, E58P, E58V, E58Y, and E58W; (8) An amino acid substitution at the Gly59 position in CDR2β selected from G59P, G59L, G59D, G59E, G59H, G59K, G59Q, G59R, G59S, and G59T; (9) An amino acid substitution at the Ser63 position in CDR2β selected from S63D, S63E, S63H, S63K, S63Q, S63R, and S63T; (10) An amino acid substitution at the Lys64 position in CDR2β selected from K64Q, K64M, K64C, K64W, K64T, K64P, and K64A; (11) An amino acid substitution at the Ala65 position in CDR2β selected from A65F, A65I, A65L, A65M, A65P, A65V, A65W, and A65Y; (12) S105L, S105N, S105H, S105Q, S105K, and S105 R selected from, an amino acid substitution at the Ser105 position in CDR3β; (13) An amino acid substitution at the Glu108 position in CDR3β selected from E108D, E108F, E108T, E108I, E108K, and E108P; (14) The amino acid substitution G110W at the Gly110 position in CDR3β; (15) An amino acid substitution at the Gln111 position in CDR3β selected from Q111D, Q111N, Q111K, Q111S, Q111G, Q111A, Q111V, Q111E, Q111L, and Q111H; (16) An amino acid substitution of EPGQG to VNDGT at positions from Glu108 to Pro113 in CDR3β; (17) An amino acid substitution at the Gly112 position in CDR3β selected from G112P, G112A, and G112E; (18) An amino acid substitution at the Pro113 position in CDR3β selected from P113W and P113H; (19) An amino acid substitution at the Tyr114 position in CDR3β selected from Y114I, Y114F, Y114K, Y114D, Y114L, Y114W, Y114N, Y114P, and Y114S; (20) An amino acid substitution at the Glu115 position in CDR3β selected from E115D and E115I; (21) An amino acid substitution at the Asp27 position in CDR1α selected from D27C and D27Y; (22) An amino acid substitution at the Ser28 position in CDR1α selected from S28M, S28A, S28G, S28Y and S28P; (23) An amino acid substitution at the Ser29 position in CDR1α selected from S29E, S29T, S29Y, S29A, S29F, S29I, S29L, S29M, S29P, S29V, S29W and S29N; (24) An amino acid substitution at the Ser36 position in CDR1α selected from S36D, S36M, S36L, S36Q, S36I, S36F, S36P and S36V; (25) An amino acid substitution T37I at the Thr37 position in CDR1α; (26) An amino acid substitution Y38L at the Tyr38 position in CDR1α; (27) An amino acid substitution at the Ile56 position in CDR2α selected from I56T and I56G; (28) An amino acid substitution at the Phe57 position in CDR2α selected from F57W and F57T; (29) An amino acid substitution at the Ser58 position in CDR2α selected from S58G, S58I, S58N, S58A, S58P and S58W; (30) An amino acid substitution at the Asn59 position in CDR2α selected from N59L, N59F, N59D, N59E, N59H, N59K, N59Q, N59R, N59S and N59T; (31) An amino acid substitution at the Met63 position in CDR2α selected from M63D, M63Q, M63T and M63E; (32) An amino acid substitution at the Asp64 position in CDR2α selected from D64A, D64Q, D64T, D64Y and D64S; (33) An amino acid substitution at the Thr26 position in FR1 selection selected from T26S, T26I, T26P, T26Q, T26A and T26V; The modified TCR according to claim 1, comprising one or more of the above.

3. The modified TCR has the following amino acid substitutions (1)-(4) in CDR1β compared to the parental TCR: (1) Q29E or Q29P; (2) A36I; (3) T37E; (4) T38R; including one or more of the above, Preferably, the modified TCR according to claim 1 or 2 comprises an amino acid substitution of Q29E or Q29PA36IT37ET38R in CDR1β as compared to the parental TCR.

4. The modified TCR comprises, as compared to the parental TCR, one or more of the following amino acid substitutions (1) to (7) in CDR2β: (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; of which one or more are included, Preferably, the modified TCR according to any one of claims 1 to 3 comprises an amino acid substitution of N57DE58YG59QS63TK64CA65V, S56TE58LG59P, S56TN57QE58PG59L, S56TN57DE58Y or S63DK64Q in CDR2β as compared to the parental TCR.

5. The modified TCR comprises, as compared to the parental TCR, one or more of the following amino acid substitutions (1) to (8) in CDR3β: (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) Substitution of EPGQGP with VNDGT at positions from Glu108 to Pro113; of which one or more are included, Preferably, the modified TCR according to any one of claims 1 to 4 comprises an amino acid substitution of: G110WQ111DG112PP113W, G112AP113WY114SE115D, Y114F, Y114W, Q111GY114F, Q111SY114F, S105L, S105N, substitution of EPGQGP with VNDGT at positions from Glu to Pro113 or G112EP113HY114LE115I in CDR3β as compared to the parental TCR.

6. The modified TCR comprises, as compared to the parental TCR, the following amino acid substitutions (1) to (3): (1) An amino acid substitution of Q29E or Q29PA36IT37ET38R in CDR1β; (2) Amino acid substitutions in CDR2β: N57D E58Y G59Q S63T K64C A65V, S56T E58L G59P, S56T N57Q E58P G59L, S56T N57D E58Y or S63D K56Q; (3) Amino acid substitutions in CDR3β: G110W Q111D G112P P113W, G112A P113W Y114S E115D, Y114F, Y114W, Q111G Y114F, Q111S Y114F, S105L, S105N, substitution of EPGQG P with VNDGT at the position from Glu108 to Pro113 or G112E P113H Y114L E115I; (4) The modified TCR according to any one of claims 1 to 5, comprising one or more of the above. (5) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (6) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (7) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (8) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (9) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (10) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (11) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (12) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (13) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (14) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (15) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (16) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (17) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (18) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (19) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (20) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (21) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (22) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (23) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (24) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29M. (25) The modified TCR according to any one of claims 1 to 6, preferably, compared with the parental TCR, the modified TCR comprises amino acid substitutions in CDR1α and / or FR1α: S29T S36D T37I Y38L, T26A S28Y S29L, T26V S28P S29Y, S36L, T26S S28M S29L, T26I D27C S28A S29F, S36Q, T26P S28M S29F, S36M, S29E S36F or T26Q D27Y S28G S29 Preferably, compared with the parent TCR, the modified TCR includes amino acid substitutions N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F in CDR2α, and the modified TCR according to any one of claims 1 to 7.

9. Compared with the parent TCR, the modified TCR has the following amino acid substitutions (1) to (2): (1) Amino acid substitutions contained in CDR1α and / or FR1α: S29TS36DT37IY38L, T26AS28YS29L, T26VS28PS29Y, S36L, T26SS28MS29L, T26ID27CS28AS29F, S36Q, T26PS28MS29F, S36M, S29ES36F or T26QD27YS28GS29M; (2) Amino acid substitutions contained in CDR2α: N59F, S58IN59FM63QD64A, S58AN59FD64Q, S58IN59FM63QD64A, S58GN59FM63DD64T, S58IN59FM63QD64Y, S58NN59TM63Q, N59FM63TD64Y, S58AN59FM63ED64S, I56TF57WS58PN59H, I56GF57TS58WN59L or I56TS58WN59F; The modified TCR according to any one of claims 1 to 8, which includes one or more of the above.

10. The modified TCR is (i) an α-chain variable region including CDR1α, CDR2α and CDR3α with amino acid sequences shown in SEQ ID NOs: 27 to 29 respectively, and (ii) a β-chain variable region, CDR1β, CDR2β and CDR3β with amino acid sequences shown in SEQ ID NOs: 24, 30 and 26 respectively, CDR1β, CDR2β and CDR3β with amino acid sequences shown in SEQ ID NOs: 24, 25 and 31 respectively, CDR1β, CDR2β and CDR3β with amino acid sequences shown in SEQ ID NOs: 24, 25 and 32 respectively, CDR1β, CDR2β and CDR3β with 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 NO: 24, 25, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 88, and 26, respectively, CDR1β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 26, respectively, CDR2β and CDR3β, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 94, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 25, and 131, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 153, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 162, respectively, or a modified TCR according to claim 1, comprising a β-chain variable region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 91, and 33, respectively. **Claim 11** The modified TCR is (i) a β-chain variable region comprising CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24 to 26, respectively, and (ii) an α-chain variable region, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 42, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 97, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 100, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 106, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 109, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, CDR1α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively , CDR2α, and CDR3α, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 140, 141, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 146, 118, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 149, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 76, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 165, 118, and 29, respectively, CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, or CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 141, and 29, respectively, The CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 170, 76, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 118, and 29, respectively, or the modified TCR according to claim 1, comprising a variable α-chain region containing the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149, and 29, respectively. **Claim 12** The modified TCR is (i) a variable α-chain region, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 42, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 76, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 97, 28, and 29, respectively, the CDR1α , CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 100, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 28, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 106, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 109, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 112, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 115, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118, and 29, respectively, the CDR1α, CDR2α, and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 121, and 29, respectively. CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 124 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 127 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 103, 76 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 44, 76 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 140, 141 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 146, 118 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 149 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 76 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 28 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 165, 118 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 27, 118 and 29 respectively, or CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 45, 141 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 170, 76 and 29 respectively, CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 43, 118 and 29 respectively, or a variable α-chain region comprising CDR1α, CDR2α and CDR3α of the amino acid sequences shown in SEQ ID NO: 152, 149 and 29 respectively, and (ii) a variable β-chain region, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30 and 26 respectively, CDR1β, CDR2β and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25 and 31 respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 32, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 33, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 34, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 36, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 30, and 37, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 38, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 39, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 40, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 41, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 79, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 25, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 85, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 88, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 25, and 94, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 25, and 131, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 24, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 153, and 35, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 26, respectively, CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 130, 91, and 162, respectively, or The modified TCR according to claim 1, comprising a β-chain variable region containing CDR1β, CDR2β, and CDR3β of the amino acid sequences shown in SEQ ID NO: 82, 91, and 33, respectively.

13. An α-chain variable region having the amino acid sequence 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 SEQ ID NO: Any one of the amino acid sequences shown in 48 - 64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, or SEQ ID NO: A β-chain variable region having an amino acid sequence selected from the amino acid sequences having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any one of 48 - 64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175. The modified TCR according to claim 1.

14. The α-chain variable region having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 65 to 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 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 to 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 the β-chain variable region having the amino acid sequence shown 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. The modified TCR according to claim 1.

15. The amino acid sequence shown in any one of SEQ ID NOs: 65 to 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 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, 132, 136, 142, 147, 150, 154, 158, 166, 168, 171, 173, and an amino acid sequence shown in any one of SEQ ID NOs: 48 to 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 to 64, 80, 83, 86, 89, 92, 95, 133, 137, 143, 155, 159, 163, 175, the modified TCR according to claim 1, comprising a β-chain variable region having an amino acid sequence selected from the group consisting of:

16. The modified TCR according to any one of claims 1 to 15, further comprising a TCR constant region.

17. The modified TCR according to claim 16, wherein the TCR constant region is a mouse constant region or a human constant region.

18. 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 includes at least one cysteine substitution with respect to the wild-type sequence in order to form a disulfide bond between the TCR α-chain and the TCR β-chain, Preferably, the TCR α-chain constant region includes an LVL substitution or an LIV substitution such that the α-chain constant region includes the amino acid sequence LLIVILRL, the modified TCR according to claim 16 or 17.

19. A TCR α chain having the amino acid sequence 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 SEQ ID NOs: 3 to 19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 An amino acid sequence selected from the amino acid sequences shown in any one of the above, 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 to 19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176 The modified TCR according to claim 16, comprising a TCR β chain having an amino acid sequence.

20. A TCR α chain having the amino acid sequence shown in any one of SEQ ID NOs: 20 to 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 SEQ ID NOs: 20 to 23, 78, 99, 102, 105, 108, 111, 114, 117, 120, 123, 126, 129, 134, 138, 144, 148, 151, 156, 160, 167, 169, 172, 174 An amino acid sequence selected from amino acid sequences having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to any one of the above, and SEQ ID NO: 70, or SEQ ID NO: 70 An amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to the modified TCR according to claim 16, comprising a TCR β chain having an amino acid sequence.

21. The TCR α chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 20 to 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 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 to 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 the TCR β chain having an amino acid sequence selected from the amino acid sequences shown in any one of SEQ ID NOs: 3 to 19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176, or an amino acid sequence having at least also 80%, at least 85%, at least 90%, at least 95% or at least 99% sequence identity to any one of SEQ ID NOs: 3 to 19, 81, 84, 87, 90, 93, 96, 135, 139, 145, 157, 161, 164, 176, the modified TCR according to claim 16.

22. The modified TCR according to any one of claims 1 to 21, which binds to an epitope comprising the amino acid sequence KVLEYVIKV (SEQ ID NO: 2) or a complex of said epitope and an MHC molecule.

23. The modified TCR according to claim 22, wherein the MHC molecule is 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, which is soluble or membrane-bound.

25. The modified TCR according to any one of claims 1 to 24, which is isolated or purified.

26. A nucleic acid encoding the modified TCR according to any one of claims 1 to 25.

27. A vector containing the nucleic acid according to claim 26.

28. The vector according to claim 27, wherein the vector is selected from 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 the modified TCR according to any one of claims 1 to 25, the nucleic acid according to claim 26, or the vector according to claim 27 or 28.

30. The cell according to claim 29, which is selected from lymphocytes (e.g., T cells, NK cells), monocytes (e.g., PBMC), and stem cells.

31. The cell according to claim 30, wherein the stem cell is a lymphocyte 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 an endogenous TCR.

34. A method for producing the cell according to any one of claims 29 to 33, the method 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 proliferating and / or activating the cell before or after the transduction or transfection.

36. A conjugate comprising the modified TCR according to any one of claims 1 to 25 and an active agent bound or conjugated to the TCR.

37. The active agent is selected from a detectable label, an immune stimulatory molecule, and a therapeutic agent, preferably, the detectable label is selected from biotin, streptavidin, an enzyme or a catalytic 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 immune stimulatory molecule is selected from cytokines, chemokines, platelet factors, and complement promoters, preferably, the therapeutic agent is selected from immunomodulators, radioactive compounds, enzymes, chemotherapeutic agents, and toxins. The conjugate according to claim 36.

38. A bispecific or multispecific antigen-binding molecule comprising a modified TCR according to any one of claims 1 to 25 or an antigen-binding fragment thereof and one or more additional antigen-binding regions that bind to 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, a bispecific or multispecific antigen-binding molecule.

39. The bispecific or multispecific antigen-binding molecule according to claim 38, wherein 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.

40. The bispecific or multispecific antigen-binding molecule according to claim 39, wherein the one or more additional antigens are CD3, and preferably the one or more additional antigen-binding regions are derived from the antigen-binding region of an anti-CD3 antibody.

41. 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, a composition further comprising a pharmaceutically acceptable carrier or excipient.

42. The composition according to claim 41, further comprising a second therapeutic agent, and preferably the second therapeutic agent is selected from an antibody, a chemotherapeutic agent, and a small molecule drug.

43. A method of treating 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 to 25.

44. A method of treating 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 to 33.

45. The method according to claim 44, wherein the cell is autologous or allogeneic to the subject.

46. (i) isolating a cell-containing sample from the subject; (ii) transducing or transfecting the cell with the vector according to claim 27 or 28; (iii)administering the cells obtained in step (ii), the method according to claim 44, comprising: **Claim 47** The method according to claim 46, further comprising, after step (i) and before (ii), knocking out the endogenous TCR in said cells. **Claim 48** The MAGE-A1 positive cancer is melanoma, lung cancer, colorectal cancer, cervical cancer, breast cancer, liver cancer (e.g., hepatocellular carcinoma), gastric cancer, esophageal cancer, bladder cancer (e.g., bladder urothelial carcinoma), ovarian cancer (e.g., ovarian serous cystadenocarcinoma), head and neck cancer (e.g., head and neck squamous cell carcinoma), uterine cancer, endometrial cancer, cholangiocarcinoma, prostate cancer, adrenocortical carcinoma, mesothelioma, sarcoma, pheochromocytoma, paraganglioma, and thymic carcinoma, the method according to any one of claims 43 to 47. **Claim 49** Further comprising administering a second therapeutic agent, preferably, the second therapeutic agent is selected from antibodies, chemotherapeutic agents, and small molecule drugs, the method according to any one of claims 43 to 48. **Claim 50** The subject has an HLA-A*02 allele, e.g., an HLA-A*02:01 or HLA-A*02:05 allele, the method according to any one of claims 43 to 49.

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