Pharmaceutical composition for treating or preventing cancer

Abstract

As a technique that can be used in immunotherapy against KitaKyushu Lung Cancer Antigen 1 (KK-LC-1)-expressing cancer, a pharmaceutical composition is provided which is administered in combination in order to treat or prevent KK-LC-1-expressing cancer in a subject: a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and a T cell, the peptide and the T cell having the following combination according to the human leukocyte antigen (HLA) type of the subject: (1) a combination of a peptide comprising the amino acid sequence of SEQ ID NO: 1 and a T cell expressing a T cell receptor (TCR) having a beta chain comprising the complementarity determining region 3 (CDR3) of SEQ ID NO: 2 when the HLA type of the subject is HLA * C14, and (2) a combination of a peptide comprising the amino acid sequence of SEQ ID NO: 1 and a T cell expressing a T cell receptor (TCR) having a beta chain comprising the complementarity determining region 3 (CDR3) of SEQ ID NO: 2 when the HLA type of the subject is HLA * C14; or (2) a combination of a peptide comprising the amino acid sequence of SEQ ID NO: 3 and a T cell expressing a TCR having a beta chain comprising CDR3 of SEQ ID NO: 4, 5 or 6 when the HLA type of the subject is HLA * B15.

Description

Technical Field

[0001] The present disclosure relates to a pharmaceutical composition for treating or preventing cancer. More specifically, it relates to a pharmaceutical composition for treating or preventing cancer expressing KitaKyushu Lung Cancer Antigen 1 (KK-LC-1), which is administered in combination with a peptide or the like and T cells. Background Art

[0002] KitaKyushu Lung Cancer Antigen 1 (KK-LC-1) is a cancer antigen expressed in various cancers such as lung cancer, gastric cancer, breast cancer, and liver cancer, and is expressed only in the testis in normal tissues. A cancer immunotherapy reagent using a peptide derived from KK-LC-1 is disclosed in Patent Document 1.

[0003] Prior Art Documents

[0004] Patent Documents

[0005] Patent Document 1: International Publication No. 2017 / 089756 Summary of the Invention

[0006] Problems to be Solved by the Invention

[0007] The main object of the present disclosure is to provide a technique that can be used for immunotherapy against cancer expressing KK-LC-1.

[0008] Means for Solving the Problems

[0009] To solve the above problems, the present disclosure provides the following [1] -

[23] .

[0010] [1] A pharmaceutical composition that is administered in combination for treating or preventing cancer expressing KK-LC-1 in a subject:

[0011] A peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and

[0012] T cells,

[0013] The aforementioned peptide and the aforementioned T cells have the following combination of (1) or (2) according to the human leukocyte antigen (HLA) type possessed by the aforementioned subject.

[0014] (1) When the HLA type of the aforementioned subject is HLA*C14, it is a combination of the following peptide and the following T cells, the peptide contains the amino acid sequence of SEQ ID NO: 1, and the T cell expresses a T cell receptor (TCR) having a β chain containing the complementary determining region 3 (CDR3) of SEQ ID NO: 2.

[0015] (2) When the HLA type of the aforementioned subject is HLA*B15, it is a combination of the following peptide and the following T cells, the peptide contains the amino acid sequence of SEQ ID NO: 3, and the T cells express a TCR with a β chain having a CDR3 containing SEQ ID NO: 4, 5, or 6.

[0016] [2] A pharmaceutical composition, which is administered in combination for the treatment or prevention of cancer expressing KK-LC-1 in a subject:

[0017] A peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and

[0018] T cells,

[0019] The aforementioned peptide and the aforementioned T cells have the following combination of (1) or (2).

[0020] (1) A combination of the following peptide and the following T cells, the peptide contains the amino acid sequence of SEQ ID NO: 1, and the T cells express a TCR with a β chain having a CDR3 containing SEQ ID NO: 2.

[0021] (2) A combination of the following peptide and the following T cells, the peptide contains the amino acid sequence of SEQ ID NO: 3, and the T cells express a TCR with a β chain having a CDR3 containing SEQ ID NO: 4, 5, or 6.

[0022] [3] The pharmaceutical composition according to [2], wherein the HLA type of the aforementioned subject is HLA*C14 in the combination of the aforementioned (1),

[0023] and is HLA*B15 in the combination of the aforementioned (2).

[0024] [4] The pharmaceutical composition according to [3], wherein the HLA type of the aforementioned subject is HLA*C14:02 in the combination of the aforementioned (1),

[0025] and is HLA*B15:01 in the combination of the aforementioned (2).

[0026] [5] The pharmaceutical composition according to any one of [1]-[4], wherein the aforementioned cancer is lung cancer, gastric cancer, breast cancer, or liver cancer.

[0027] [6] A pharmaceutical composition, which contains a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, which is administered in combination with T cells for the treatment or prevention of cancer expressing KK-LC-1 in a subject,

[0028] The aforementioned peptide is the following (1) or (2).

[0029] (1) A peptide containing the amino acid sequence of SEQ ID NO: 1, which is administered in combination with T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 2.

[0030] (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3, administered in combination with T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

[0031] [7] A pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, the pharmaceutical composition comprising a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide.

[0032] The aforementioned peptide is one of the following (1) or (2).

[0033] (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0034] (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0035] [8] The pharmaceutical composition according to [6] or [7], which is a cancer vaccine.

[0036] [9] The pharmaceutical composition according to any one of [6]-[8], wherein the aforementioned cancer is lung cancer, gastric cancer, breast cancer, or liver cancer.

[0037]

[10] A peptide comprising the amino acid sequence of SEQ ID NO: 1 or 3, a polynucleotide encoding the peptide, or a vector containing the polynucleotide.

[0038]

[11] A pharmaceutical composition comprising T cells administered in combination with a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide for treating or preventing cancer expressing KK-LC-1 in a subject.

[0039] The aforementioned T cells are one of the following (1) or (2).

[0040] (1) T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 2, administered in combination with a peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0041] (2) T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6, administered in combination with a peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0042]

[12] A pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, the pharmaceutical composition comprising T cells.

[0043] The aforementioned T cells are one of the following (1) or (2).

[0044] (1) T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 2.

[0045] (2) T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

[0046]

[13] A pharmaceutical composition as in

[11] or

[12] , wherein the cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0047]

[14] A polynucleotide, or a vector containing the polynucleotide, the polynucleotide encoding a TCR having a β-chain with a CDR3 comprising SEQ ID NO: 2, 4, 5 or 6.

[0048]

[15] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising the step of administering a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, in combination with T cells, to the subject.

[0049] The aforementioned peptide and the aforementioned T cells have the following combination of (1) or (2) according to the human leukocyte antigen (HLA) type of the aforementioned subject.

[0050] (1) A combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 1, the T cells expressing a TCR having a β-chain with a CDR3 comprising SEQ ID NO: 2.

[0051] (2) A combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 3, the T cells expressing a TCR having a β-chain with a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0052]

[16] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising:

[0053] The step of determining the HLA type of the aforementioned subject; and

[0054] The step of administering a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, in combination with T cells.

[0055] (1) When the HLA type of the aforementioned subject is HLA*C14, administering a peptide comprising the amino acid sequence of SEQ ID NO: 1, in combination with T cells expressing a TCR having a β-chain with a CDR3 comprising SEQ ID NO: 2.

[0056] (2) When the HLA type of the aforementioned subject is HLA*B15, administering a peptide comprising the amino acid sequence of SEQ ID NO: 3, in combination with T cells expressing a TCR having a β-chain with a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0057]

[17] The method as in

[15] or

[16] , wherein the HLA type of the aforementioned subject is HLA*C14:02 in the aforementioned (1),

[0058] and HLA*B15:01 in the aforementioned (2).

[0059]

[18] The method according to any one of

[15] -

[17] , wherein the cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0060]

[19] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising the step of administering to the subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide,

[0061] The peptide is one of the following (1) or (2).

[0062] (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0063] (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0064]

[20] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising:

[0065] The step of determining the HLA type of the subject; and

[0066] The step of administering to the subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide,

[0067] The peptide is one of the following (1) or (2).

[0068] (1) When the HLA type of the subject is HLA*C14, a peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0069] (2) When the HLA type of the subject is HLA*B15, a peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0070]

[21] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising the step of administering T cells to the subject,

[0071] The T cells are one of the following (1) or (2).

[0072] (1) T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2.

[0073] (2) T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0074]

[22] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising:

[0075] The step of determining the HLA type of the subject; and

[0076] The step of administering T cells to the subject,

[0077] The aforementioned T cells are as follows (1) or (2).

[0078] (1) When the HLA type of the aforementioned subject is HLA*C14, T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 2.

[0079] (2) When the HLA type of the aforementioned subject is HLA*B15, T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 4, 5, or 6.

[0080] Use of a

[23] -peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, or T cells for the manufacture of a pharmaceutical composition for the treatment or prevention of cancer expressing KK-LC-1 in a subject.

[0081] The aforementioned pharmaceutical composition has a combination of the aforementioned peptide and the aforementioned T cells as follows (1) or (2).

[0082] (1) A combination of a peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a T cell receptor (TCR) with a β chain having a CDR3 containing SEQ ID NO: 2.

[0083] (2) A combination of a peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 4, 5, or 6.

[0084] In the present disclosure, "immunogenicity" means the ability to induce an immune response (immune induction ability). For example, it means increasing the cytotoxic T lymphocyte (CTL) induction activity of antigen-presenting cells, and further increasing the cytotoxic activity of CTL against cancer cells.

[0085] In addition, in the present disclosure, "CTL induction" means that in vitro or in vivo, the peptide involved in the present disclosure is presented on the surface of antigen-presenting cells, thereby inducing or proliferating CTLs that specifically recognize the KK-LC-1 antigen; or differentiating naive T cells into effector cells having the ability (cytotoxic activity) to kill target cells such as cancer cells; and / or increasing the cytotoxic activity of CTLs.

[0086] CTL-induced activity can be measured by evaluating cytokines (such as IFN-γ) produced by CTLs. For example, the increase in cytokine-producing cells induced from progenitor cells by antigen-presenting cells such as peripheral blood mononuclear cells (stimulated with the peptides involved in the present disclosure) can be evaluated using known highly sensitive immunoassay methods such as the ELISPOT (Enzyme-Linked ImmunoSpot) method or the ELISA (Enzyme-Linked ImmunoSorbent Assay) method, thereby measuring CTL-induced activity.

[0087] The cytotoxic activity of CTLs can also be measured using 51 known methods such as the Cr release method.

[0088] When the CTL-induced activity of the test substance is significantly increased compared to the control substance, for example, increased by 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, preferably 50% or more, it can be determined that the test substance has CTL-induced activity.

[0089] Advantages of the Invention

[0090] Through the present disclosure, a technique that can be used for immunotherapy against cancers expressing KK-LC-1 is provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0091] Figure 1 shows the detection results of antigen-specific reactions in the co-culture of B-APCs loaded with KK-LC-1 peptides and TexTCR-expressing cells expressing TexTCR (pt.2-ex08TCR).

[0092] Figure 2 shows the detection results of antigen-specific reactions in the co-culture of HLA-expressing cells loaded with the peptide CT83-12 (RFQRNTGEM) and TexTCR-expressing cells expressing TexTCR (pt.2-ex08TCR).

[0093] Figure 3 shows the detection results of antigen-specific reactions in the co-culture of B-APCs loaded with KK-LC-1 peptides and TexTCR-expressing cells expressing TexTCR (pt.1-ex08TCR, pt.1-ex15TCR or pt.2-ex09TCR).

[0094] Figure 4 shows the full-length amino acid sequences of the β-chain and α-chain and the CDRs of TexTCR (pt.2-ex08TCR). ​​​​

[0095] Figure 5 represents the full-length amino acid sequences of the β-chain and α-chain of TexTCR (pt.1-ex08TCR) and the CDRs.

[0096] Figure 6 represents the full-length amino acid sequences of the β-chain and α-chain of TexTCR (pt.1-ex15TCR) and the CDRs.

[0097] Figure 7 represents the full-length amino acid sequences of the β-chain and α-chain of TexTCR (pt.2-ex09TCR) and the CDRs. Detailed implementation modes

[0098] [Peptide]

[0099] The peptide involved in the present disclosure text contains the amino acid sequence of SEQ ID NO: 1 or 3.

[0100] The peptide involved in the present disclosure text is presented by HLA of antigen-presenting cells and thus recognized by the TCR of T cells. Therefore, the peptide involved in the present disclosure text can be effectively used for immunotherapy against cancers expressing KK-LC-1.

[0101] The amino acid sequence of SEQ ID NO: 1 corresponds to positions 23-31 from the N-terminus of the full-length amino acid sequence of KK-LC-1. The peptide containing the amino acid sequence of SEQ ID NO: 1 has HLA*C14 restriction, particularly HLA*C14:02 restriction. Therefore, the peptide containing the amino acid sequence of SEQ ID NO: 1 can be suitably used for CTL induction in subjects with HLA type HLA*C14, particularly HLA*C14:02.

[0102] The amino acid sequence of SEQ ID NO: 3 corresponds to positions 76-84 from the N-terminus of the full-length amino acid sequence of KK-LC-1. The peptide containing the amino acid sequence of SEQ ID NO: 3 has HLA*B15 restriction, particularly HLA*B15:01 restriction. Therefore, the peptide containing the amino acid sequence of SEQ ID NO: 3 can be suitably used for CTL induction in subjects with HLA type HLA*B15, particularly HLA*B15:01.

[0103] The peptide involved in the present disclosure text may contain all or a part of the amino acid sequence of SEQ ID NO: 1 or 3, and for being presented by HLA, the total number of its full-length amino acids is preferably set to 8-11.

[0104] The peptide involved in the present disclosure text preferably comprises the whole of the amino acid sequence of SEQ ID NO: 1 or 3, and particularly preferably consists of the amino acid sequence of SEQ ID NO: 1 or 3.

[0105] ​​​Regarding the peptides involved in the present disclosure, the amino acid residues or a part thereof that constitute the amino acid sequence may be modified. The modification is not particularly limited, and examples thereof include sugar chain addition, side chain oxidation, and phosphorylation.

[0106] In addition, as long as the peptides involved in the present disclosure maintain immunogenicity, one or several amino acid residues may also be substituted, inserted, deleted, or added. When the peptide is directly administered into the body, depending on the administration route, changes such as digestion of the peptide terminus in the digestive organs may occur. Therefore, the peptides involved in the present disclosure may exist in the form of a precursor obtained by adding one or more amino acid residues, etc. to the N-terminus and / or C-terminus.

[0107] The peptides involved in the present disclosure may be replaced by polynucleotides encoding the peptides, vectors containing the polynucleotides, or antigen-presenting cells presenting the complex of the peptide and HLA molecules on the surface. The peptides involved in the present disclosure induce CTLs by being presented by HLA of antigen-presenting cells, and the induced CTLs kill cancer cells. By administering these polynucleotides, vectors, and antigen-presenting cells, CTLs can also be induced in the same manner as the administration of the peptides.

[0108] [T cell]

[0109] The T cells involved in the present disclosure express a TCR having a β chain containing a CDR3 of any one of SEQ ID NOs: 2, 4-6.

[0110] In the binding of the complex formed by the HLA molecule and the antigen peptide presented by it to the TCR, CDR3 is involved in the recognition of the antigen peptide, and the binding property to the antigen peptide is particularly determined by the CDR3 of the β chain.

[0111] The TCR of the T cells involved in the present disclosure recognizes the KK-LC-1 peptide presented by HLA. Therefore, the T cells involved in the present disclosure can be effectively used for immunotherapy against cancers expressing KK-LC-1.

[0112] T cells expressing a TCR having a β chain containing a CDR3 of SEQ ID NO: 2 can bind to HLA molecules presenting a peptide containing the amino acid sequence of SEQ ID NO: 1. The peptide containing the amino acid sequence of SEQ ID NO: 1 has HLA*C14 (especially HLA*C14:02) restriction. Therefore, T cells expressing a TCR having a β chain containing a CDR3 of SEQ ID NO: 2 are preferably administered in combination with the peptide containing the amino acid sequence of SEQ ID NO: 1, and more preferably administered together with the peptide to a subject having an HLA type of HLA*C14 (especially HLA*C14:02).

[0113] In the case of T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 2, as an example, there may be mentioned those further comprising: an HLA-binding region (CDR1) of SEQ ID NO: 17 and an HLA-binding region (CDR2) of SEQ ID NO: 18 as the complementarity-determining regions of the β-chain; an HLA-binding region (CDR1) of SEQ ID NO: 19, an HLA-binding region (CDR2) of SEQ ID NO: 20, and an HLA-binding region (CDR3) of SEQ ID NO: 21 as the complementarity-determining regions of the α-chain.

[0114] T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 2 may contain the amino acid sequence of SEQ ID NO: 22 as the full-length sequence of the β-chain and the amino acid sequence of SEQ ID NO: 23 as the full-length sequence of the α-chain.

[0115] T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 4, 5, or 6 can bind to an HLA molecule presenting a peptide containing the amino acid sequence of SEQ ID NO: 3. The peptide containing the amino acid sequence of SEQ ID NO: 3 has HLA-B15 (particularly HLA-B15:01) restriction. Therefore, T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 4, 5, or 6 are preferably administered in combination with a peptide containing the amino acid sequence of SEQ ID NO: 3, and more preferably administered together with the peptide to a subject with an HLA type of HLA-B15 (particularly HLA-B15:01).

[0116] In the case of T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 4, as an example, there may be mentioned those further comprising: an HLA-binding region (CDR1) of SEQ ID NO: 24 and an HLA-binding region (CDR2) of SEQ ID NO: 25 as the complementarity-determining regions of the β-chain; an HLA-binding region (CDR1) of SEQ ID NO: 26, an HLA-binding region (CDR2) of SEQ ID NO: 27, and an HLA-binding region (CDR3) of SEQ ID NO: 28 as the complementarity-determining regions of the α-chain.

[0117] In the case of T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 4, they may contain the amino acid sequence of SEQ ID NO: 29 as the full-length sequence of the β-chain and the amino acid sequence of SEQ ID NO: 30 as the full-length sequence of the α-chain.

[0118] In the case of T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 5, as an example, there may be mentioned those further comprising: an HLA-binding region (CDR1) of SEQ ID NO: 31 and an HLA-binding region (CDR2) of SEQ ID NO: 32 as the complementarity-determining regions of the β-chain; an HLA-binding region (CDR1) of SEQ ID NO: 33, an HLA-binding region (CDR2) of SEQ ID NO: 34, and an HLA-binding region (CDR3) of SEQ ID NO: 35 as the complementarity-determining regions of the α-chain.

[0119] Regarding T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 5, it may include the amino acid sequence of SEQ ID NO: 36 as the full-length sequence of the β-chain and the amino acid sequence of SEQ ID NO: 37 as the full-length sequence of the α-chain.

[0120] Regarding T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 6, as an example, it may further include: the HLA-binding region (CDR1) of SEQ ID NO: 38 and the HLA-binding region (CDR2) of SEQ ID NO: 39 as the complementarity-determining regions of the β-chain; the HLA-binding region (CDR1) of SEQ ID NO: 40, the HLA-binding region (CDR2) of SEQ ID NO: 41, and the HLA-binding region (CDR3) of SEQ ID NO: 42 as the complementarity-determining regions of the α-chain.

[0121] Regarding T cells expressing a TCR with a β-chain containing a CDR3 of SEQ ID NO: 6, it may include the amino acid sequence of SEQ ID NO: 43 as the full-length sequence of the β-chain and the amino acid sequence of SEQ ID NO: 44 as the full-length sequence of the α-chain.

[0122] The T cells related to the present disclosure may be T cells (TCR gene-transferred T cells) that express the above-mentioned TCR by gene transfer.

[0123] Using a vector containing a polynucleotide encoding a TCR, peripheral blood lymphocytes derived from a subject in need of treatment or prevention of cancer are transformed, and after the resulting TCR gene-transferred T cells are proliferated, they are returned to the subject's body.

[0124] The polynucleotide encoding a TCR or a vector containing the same can be introduced into peripheral blood lymphocytes by a conventionally known method such as liposome transfection, electroporation, microinjection, cell fusion, DEAE-dextran method, calcium phosphate method, etc.

[0125] In addition, the T cells related to the present disclosure can be replaced by T cells expressing a CAR (CAR-T), and the CAR contains an antigen recognition region derived from an antibody that binds to the complex formed by the peptide related to the present disclosure and an HLA molecule. Usually, the antigen recognition region uses scFv.

[0126] [Pharmaceutical composition]

[0127] The pharmaceutical composition related to the present disclosure is a pharmaceutical composition in which the above-mentioned peptide, the polynucleotide encoding the peptide, or the vector containing the polynucleotide is administered in combination with T cells, and the aforementioned peptide and the aforementioned T cells have the following combination of (1) or (2).

[0128] (1) The combination of the following peptide and the following T cells, the peptide contains the amino acid sequence of SEQ ID NO: 1, and the T cells express a TCR with a β-chain containing a CDR3 of SEQ ID NO: 2.

[0129] (2) The combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 3, and the T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

[0130] In addition, in another aspect, the pharmaceutical composition according to the present disclosure may contain only the peptide described in (1) or (2) above or only T cells.

[0131] The peptide comprising the amino acid sequence of SEQ ID NO: 1 is presented by an HLA molecule of HLA*C14 (particularly HLA*C14:02) and recognized by T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 2. Therefore, the peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 2, when administered in combination to a subject having an HLA type of HLA*C14 (particularly HLA*C14:02), can be particularly effectively used for the treatment or prevention of cancer expressing KK-LC-1.

[0132] The peptide comprising the amino acid sequence of SEQ ID NO: 3 is presented by an HLA molecule of HLA-B15 (particularly HLA-B15:01) and recognized by T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6. Therefore, the peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6, when administered in combination to a subject having an HLA type of HLA-B15 (particularly HLA-B15:01), can be particularly effectively used for the treatment or prevention of cancer expressing KK-LC-1.

[0133] The cancer to which the pharmaceutical composition according to the present disclosure is applicable is not particularly limited as long as it is a cancer expressing KK-LC-1. For example, it can be lung cancer, gastric cancer, breast cancer, or liver cancer.

[0134] The pharmaceutical composition according to the present disclosure can be used not only for the treatment of cancer but also for the prevention of cancer. For example, by administering the pharmaceutical composition according to the present disclosure to a healthy human body, CTLs are induced, and the induced CTLs remain in the body. Therefore, when cancer cells are generated, they can kill the cancer cells. Similarly, it can also be administered to a human body after cancer treatment to prevent cancer recurrence.

[0135] In the pharmaceutical composition according to the present disclosure, the peptide and T cells can be formulated into the same preparation or into separate preparations. In the case where the peptide and T cells are formulated into separate preparations, the preparation containing the peptide and the preparation containing T cells can be administered to the subject simultaneously or at different times.

[0136] The dosages of the peptide and T cells can be appropriately set according to the age, body weight, administration method, etc. of the patient in a manner to obtain the target immunogenicity.

[0137] The pharmaceutical composition related to the present disclosure may contain other components known to be used in cancer treatment, such as chemokines, cytokines, tumor necrosis factors, chemotherapeutic agents, etc.

[0138] The pharmaceutical composition related to the present disclosure may contain various additives. As additives, for example, preservatives, buffers, etc. can be cited. As preservatives, sodium bisulfite, sodium hydrogen sulfate, sodium thiosulfate, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercuric acetate, phenylmercuric nitrate, methyl paraben, polyvinyl alcohol, phenethyl alcohol, ammonia, dithiothreitol, β-mercaptoethanol, etc. can be cited. In addition, as buffers, sodium carbonate, sodium borate, sodium phosphate, sodium acetate, sodium bicarbonate, etc. can be cited.

[0139] The pharmaceutical composition related to the present disclosure can be used as a cancer vaccine in a manner containing the peptide and not containing T cells.

[0140] In addition, the vaccine may contain inactive components for enhancing the effect as a vaccine. As inactive components, adjuvants, toxoids, etc. can be cited. As examples of adjuvants, precipitating type adjuvants such as aluminum hydroxide, aluminum phosphate, calcium phosphate, etc., and oily type adjuvants such as Freund's complete adjuvant, Freund's incomplete adjuvant, etc. can be cited.

[0141] The pharmaceutical composition related to the present disclosure is not only intended for administration to the human body but also for in vitro use in a manner containing the peptide and not containing T cells.

[0142] Specifically, the pharmaceutical composition related to the present disclosure can be used to increase the CTL induction activity by stimulating antigen-presenting cells in vitro or ex vivo. For example, the pharmaceutical composition related to the present disclosure can be used in cancer dendritic cell therapy, that is, after contacting it with antigen-presenting cells such as dendritic cells derived from a subject in need of treating or preventing cancer, returning the antigen-presenting cell to the subject's body.

[0143] The peptide can be introduced into antigen-presenting cells, for example, by liposome transfection method, injection method, etc. In addition, a polynucleotide encoding the peptide and a vector containing the polynucleotide can be introduced into antigen-presenting cells, for example, by liposome transfection method, electroporation method, microinjection method, cell fusion method, DEAE-dextran method, calcium phosphate method, etc., so as to express the peptide.

[0144] A vector refers to a nucleic acid construct capable of expressing mRNA, protein or polypeptide in a host cell. Known vectors such as plasmids and viruses can be used as the vector.

[0145] [Therapeutic / Preventive Methods and Companion Diagnostics]

[0146] The method for treating or preventing cancer according to the present disclosure includes the step of co-administering the above-described peptide, polynucleotide encoding the peptide, or vector containing the polynucleotide with T cells.

[0147] The aforementioned peptide and the aforementioned T cells have the combination of the following (1) or (2).

[0148] (1) The combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 1, and the T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2.

[0149] (2) The combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 3, and the T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

[0150] The administration of the peptide, polynucleotide encoding the peptide, or vector containing the polynucleotide (hereinafter, also collectively abbreviated as "peptide" for simplicity), and the administration of T cells can be administered as the same preparation or as separate preparations. The administration of the peptide and the administration of T cells can be carried out simultaneously or separately. In addition, the administration route of the peptide and the administration route of T cells can be the same or different.

[0151] In another aspect, the method for treating or preventing cancer according to the present disclosure may include the step of administering only the peptide, polynucleotide encoding the peptide, or vector containing the polynucleotide described in the above (1) or (2), or the step of administering only T cells.

[0152] The method for treating or preventing cancer according to the present disclosure may include the step of determining the HLA genotype of the subject prior to the step of administering the peptide and / or T cells. The determination of the HLA genotype can be carried out using previously known gene analysis methods.

[0153] The peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2 are administered to a subject with an HLA type, particularly HLA*C14 (especially HLA*C14:02), so as to expect an optimized therapeutic effect.

[0154] In addition, the peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6 are administered to a subject with an HLA type, particularly HLA-B15 (especially HLA-B15:01), so as to expect an optimized therapeutic effect.

[0155] Therefore, by determining the HLA genotype of a subject prior to the step of administering the peptide and / or T cell, and selecting a peptide, T cell, or a combination thereof corresponding to the HLA type of the subject, a high therapeutic effect can be expected.

[0156] Some or all of the above-described embodiments may also be as described below, but are not limited to the following. It should be noted that, for the constitution of the inventions described below, the constitution related to one category of inventions may also be subordinate to the inventions of other categories.

[0157] [1] A pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, which is administered in combination:

[0158] a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and

[0159] T cells,

[0160] The foregoing peptide and the foregoing T cells have the following combination of (1) or (2) according to the human leukocyte antigen (HLA) type possessed by the foregoing subject.

[0161] (1) When the HLA type of the foregoing subject is HLA*C14, it is a combination of the following peptide and the following T cell. The peptide contains the amino acid sequence of SEQ ID NO: 1, and the T cell expresses a T cell receptor (TCR) having a β chain containing the complementarity-determining region 3 (CDR3) of SEQ ID NO: 2.

[0162] (2) When the HLA type of the foregoing subject is HLA*B15, it is a combination of the following peptide and the following T cell. The peptide contains the amino acid sequence of SEQ ID NO: 3, and the T cell expresses a TCR having a β chain containing the CDR3 of SEQ ID NO: 4, 5, or 6.

[0163] [2] A pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, which is administered in combination:

[0164] a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and

[0165] T cells,

[0166] The foregoing peptide and the foregoing T cells have the following combination of (1) or (2).

[0167] (1) A combination of the following peptide and the following T cell. The peptide contains the amino acid sequence of SEQ ID NO: 1, and the T cell expresses a TCR having a β chain containing the CDR3 of SEQ ID NO: 2.

[0168] (2) The combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 3, the T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0169] [3] The pharmaceutical composition according to [2], wherein the HLA type of the aforementioned subject is HLA*C14 in the combination of the aforementioned (1),

[0170] and HLA*B15 in the combination of the aforementioned (2).

[0171] [4] The pharmaceutical composition according to [3], wherein the HLA type of the aforementioned subject is HLA*C14:02 in the combination of the aforementioned (1),

[0172] and HLA*B15:01 in the combination of the aforementioned (2).

[0173] [5] The pharmaceutical composition according to any one of [1]-[4], wherein the aforementioned cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0174] [6] A pharmaceutical composition comprising a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, which is administered in combination with T cells for the treatment or prevention of cancer expressing KK-LC-1 in a subject,

[0175] The aforementioned peptide is the following (1) or (2).

[0176] (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1, which is administered in combination with T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2.

[0177] (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3, which is administered in combination with T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0178] [7] A pharmaceutical composition for the treatment or prevention of cancer expressing KK-LC-1 in a subject, the pharmaceutical composition comprising a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide,

[0179] The aforementioned peptide is the following (1) or (2).

[0180] (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0181] (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0182] [8] The pharmaceutical composition according to [6] or [7], which is a cancer vaccine.

[0183] [9] The pharmaceutical composition according to any one of [6] - [8], wherein the cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0184]

[10] A peptide comprising the amino acid sequence of SEQ ID NO: 1 or 3, a polynucleotide encoding the peptide, or a vector containing the polynucleotide.

[0185]

[11] A pharmaceutical composition comprising T cells administered in combination with a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide for the treatment or prevention of cancer expressing KK-LC-1 in a subject,

[0186] The T cells are the following (1) or (2).

[0187] (1) T cells expressing a TCR β-chain having a CDR3 comprising SEQ ID NO: 2, administered in combination with a peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0188] (2) T cells expressing a TCR β-chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6, administered in combination with a peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0189]

[12] A pharmaceutical composition for the treatment or prevention of cancer expressing KK-LC-1 in a subject, the pharmaceutical composition comprising T cells,

[0190] The T cells are the following (1) or (2).

[0191] (1) T cells expressing a TCR β-chain having a CDR3 comprising SEQ ID NO: 2.

[0192] (2) T cells expressing a TCR β-chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0193]

[13] The pharmaceutical composition according to

[11] or

[12] , wherein the cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0194]

[14] A polynucleotide, or a vector containing the polynucleotide, encoding a TCR β-chain having a CDR3 comprising SEQ ID NO: 2, 4, 5 or 6.

[0195]

[15] A method for the treatment or prevention of cancer expressing KK-LC-1 in a subject, comprising the step of administering in combination to the subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide and T cells,

[0196] The peptide and the T cells have the following combination of (1) or (2) according to the human leukocyte antigen (HLA) type of the subject.

[0197] (1) The combination of the following peptide and the following T cells, wherein the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the T cells express a TCR with a β chain having a CDR3 comprising SEQ ID NO: 2.

[0198] (2) The combination of the following peptide and the following T cells, wherein the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the T cells express a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0199]

[16] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising:

[0200] The step of determining the HLA type of the aforementioned subject; and

[0201] The step of administering in combination a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide and T cells,

[0202] (1) When the HLA type of the aforementioned subject is HLA*C14, administering in combination a peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 2,

[0203] (2) When the HLA type of the aforementioned subject is HLA*B15, administering in combination a peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR with a β chain having a CDR3 comprising SEQ ID NO: 4, 5 or 6.

[0204]

[17] The method according to

[15] or

[16] , wherein the HLA type of the aforementioned subject is HLA*C14:02 in the aforementioned (1),

[0205] and HLA*B15:01 in the aforementioned (2).

[0206]

[18] The method according to any one of

[15] -

[17] , wherein the aforementioned cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0207]

[19] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising the step of administering to the aforementioned subject a peptide, a polynucleotide encoding the peptide or a vector containing the polynucleotide,

[0208] The aforementioned peptide is the following (1) or (2).

[0209] (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0210] (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3.

[0211]

[20] A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising:

[0212] The step of determining the HLA type of the aforementioned subject; and

[0213] The step of administering to the aforementioned subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide,

[0214] The aforementioned peptide is one of the following (1) or (2).

[0215] (1) When the HLA type of the aforementioned subject is HLA*C14, a peptide containing the amino acid sequence of SEQ ID NO: 1.

[0216] (2) When the HLA type of the aforementioned subject is HLA*B15, a peptide containing the amino acid sequence of SEQ ID NO: 3.

[0217]

[21] The method according to

[20] , wherein the HLA type possessed by the aforementioned subject is HLA*C14:02 in the aforementioned (1),

[0218] and HLA*B15:01 in the aforementioned (2).

[0219]

[22] A method for treating or preventing cancer expressing KK-LC-1 in a subject, which comprises the step of administering T cells to the aforementioned subject,

[0220] The aforementioned T cells are one of the following (1) or (2).

[0221] (1) T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 2.

[0222] (2) T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 4, 5, or 6.

[0223]

[23] A method for treating or preventing cancer expressing KK-LC-1 in a subject, which comprises:

[0224] The step of determining the HLA type of the aforementioned subject; and

[0225] The step of administering T cells to the aforementioned subject,

[0226] The aforementioned T cells are one of the following (1) or (2).

[0227] (1) When the HLA type of the aforementioned subject is HLA*C14, T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 2.

[0228] (2) When the HLA type of the aforementioned subject is HLA*B15, T cells expressing a TCR with a β chain having a CDR3 containing SEQ ID NO: 4, 5, or 6.

[0229]

[24] The method as described in

[23] , wherein the HLA type of the aforementioned subject is HLA*C14:02 in the aforementioned (1),

[0230] and is HLA*B15:01 in the aforementioned (2).

[0231]

[24] The method as described in

[19] -

[23] , wherein the aforementioned cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

[0232] Examples

[0233] 1. Preparation of HLA gene expression vectors

[0234] The HLA genes of HLA types A*24:02, A*26:03, B*15:01, B*51:01, C*07:02, and C*14:02 were cloned into the expression vector (pcDNA3.1(+)) to prepare the HLA gene expression vectors pcDNA3.1(+) / HLA-A*24:02, pcDNA3.1(+) / HLA-A*26:03, pcDNA3.1(+) / HLA-B*15:01, pcDNA3.1(+) / HLA-B*51:01, pcDNA3.1(+) / HLA-C*07:02, pcDNA3.1(+) / HLA-C*14:02.

[0235] 2. Introduction of HLA genes into HEK293T cells

[0236] 5 μL of Lipofectamine 2000 (Thermo Fisher Scientific) was added to OPTI-MEM medium (250 μL) and allowed to react for 5 minutes, and then mixed with OPTI-MEM medium (250 μL) containing the HLA gene expression vector (2.5 μg), and left standing for 20 minutes. 8×10 5 HEK293T cells were added to the mixture, and cultured for 6 hours. The medium was changed, and further cultured for 2 days to obtain HLA-expressing cells.

[0237] 3. Introduction of TCR genes into Jurkat cells

[0238] Using the coding sequence of a self-cleaving peptide (P2A), the cDNA of the β-chain of TCR (TexTCR) identified from tumor-infiltrating lymphocytes of a lung cancer patient was combined with the cDNA of the α-chain to prepare a synthetic gene. The synthetic gene was inserted into the retroviral vector pMX-IRES-puro (CELL BIOLABS INC) (pMX-IRES-puro / TexTCR).

[0239] Using Lipofectamine 2000, pMX-IRES-puro / TexTCR (2.5 μg) was transfected into retrovirus-producing cells and cultured. The retrovirus-producing cells used were cells obtained by introducing the GALV gene into Phoenix-GP cells (ATCC, CRL-3215).

[0240] Six hours after the start of culture, the medium was changed, and then cultured for 3 days. The culture supernatant was filtered through a 0.8-μm filter to obtain the virus solution.

[0241] Suspend 1 × 10 5 Jurkat cells that do not express TCR but express CD8 in 1 mL of the retrovirus solution. After mixing with polybrene (8 μg / ml), transfer to a 12-well plate and infect using the spin infection method (1,000 g, 1 hour, 32°C). Four hours later, add the medium at a volume of 2 ml / well. Two days after infection, collect the cells and suspend them in a new medium. Add puromycin (1 μg / ml) and culture. Two weeks later, confirm the expression of TCR using a flow cytometer to obtain Jurkat cells that express TexTCR and express CD8.

[0242] 4. KK-LC-1 peptide

[0243] Synthesize the KK-LC-1 peptide with the following amino acid sequence. For the peptide, prepare a 20 mg / mL solution using DMSO and use it after appropriate dilution.

[0244] CT83-S1: ILNNFPHSI (SEQ ID NO: 7)

[0245] CT83-S2: RQKRILVNL (SEQ ID NO: 3)

[0246] CT83-K1: SILCALIVF (SEQ ID NO: 8)

[0247] CT83-K2: IVFWKYRRF (SEQ ID NO: 9)

[0248] CT83-K3: LVRPSSSGL (SEQ ID NO: 10)

[0249] IcHi010_MP41: YCWEYFLSL (SEQ ID NO: 11)

[0250] CT83-7: VRPSSSGLI (SEQ ID NO: 12)

[0251] CT83-8: YLLLASSIL (SEQ ID NO: 13)

[0252] CT83-9: NFPHSIARQ (Serial Number 14)

[0253] CT83-10: LVELEHTLL (Serial Number 15)

[0254] CT83-11: LLLASSILC (Serial Number 16)

[0255] CT83-12: RFQRNTGEM (Serial Number 1)

[0256] 5. Detection of Antigen-Specific Reaction

[0257] The TexTCR-expressing cells were genetically modified to express luciferase under the regulation of TCR signals. Specifically, a solution (2 μg / μL) of a vector (pGL4.30[luc2P / NFAT-RE / Hygro]) containing an NFAT (nuclear factor of activated T cells) response sequence and a luciferase gene was prepared. After washing Jurkat cells expressing TexTCR and CD8 with PBS, they were suspended in electroporation buffer to prepare 2×10 5 cells / 11 μL. After mixing 1 μL of the vector solution with 11 μL of the cell solution, it was loaded onto a Neon Tip, inserted into a Neon Tube containing 3 mL of buffer, and electroporated under the conditions of 1200 V and 5 ms (Thermo Fisher Scientific, Neon transfection system).

[0258] After gene introduction, the cells were suspended in 100 μL of antibiotic-free medium.

[0259] The KK-LC-1 peptide (20 μg / ml) was loaded onto target cells and seeded in a 96-well plate (2×10 4 cells / well). Among the target cells, in addition to the HLA-expressing cells prepared in 1 above, the patient's autologous B cell antigen-presenting cells (B-APCs) were also used. The B-APCs were seeded at a quantity of 5×10 4 cells / well.

[0260] Jurkat cells expressing TexTCR and CD8 were added to each well (5×10 4After 14 hours of co-culture, a substrate solution of luciferase (Promega, Steady-Glo Luciferase Assay System) was added at 50 μL / well and reacted at room temperature for 10 minutes. 70 μL of the mixture was transferred to a half-area white plate (Greiner, LUMITRAC 675 075), and luminescence was measured using an EnSpire multimode microplate reader (PerkinElmer).

[0261] 6. Results

[0262] The detection results of antigen-specific reactions in the co-culture of peptide-loaded B-APCs and TexTCR-expressing cells expressing TexTCR (pt.2-ex08TCR) are shown in Figure 1 . Antigen-specific reactions mediated by the peptide CT83-12 (RFQRNTGEM) were detected.

[0263] The detection results of antigen-specific reactions in the co-culture of HLA-expressing cells loaded with the peptide CT83-12 (RFQRNTGEM) and TexTCR-expressing cells expressing TexTCR (pt.2-ex08TCR) are shown in Figure 2 . It was shown that the antigen-specific reaction mediated by the peptide CT83-12 (RFQRNTGEM) and TexTCR (pt.2-ex08TCR) was HLA-C14:02 restricted. Note that "None" in the figure indicates the results in EK293T cells into which the HLA gene was not introduced.

[0264] The detection results of antigen-specific reactions in the co-culture of peptide-loaded B-APCs and TexTCR-expressing cells expressing TexTCR (pt.1-ex08TCR, pt.1-ex15TCR, or pt.2-ex09TCR) are shown in Figure 3 . Regarding the peptide CT83-S2 (RQKRILVNL), antigen-specific reactions mediated by all three TexTCRs were detected. Note that the peptide CT83-S2 (RQKRILVNL) is known to be HLA-B15:01 restricted.

[0265] The combinations of HLA types, peptides, and TCRs that induce antigen-specific reactions are shown in Table 1. The full-length amino acid sequences and CDR amino acid sequences of the β-chain and α-chain of pt.2-ex08TCR, pt.1-ex08TCR, pt.1-ex15TCR, and pt.2-ex09TCR are shown in Figures 4 - 7 .

[0266] [Table 1]

[0267]

[0268] Sequence Listing Free Text

[0269] Sequence No. 1: Amino Acid Sequence of Peptide CT83-12

[0270] Sequence No. 2: Amino Acid Sequence of β-chain CDR3 of pt.2-ex08TCR

[0271] Sequence No. 3: Amino Acid Sequence of Peptide CT83-S2

[0272] Sequence No. 4: Amino Acid Sequence of β-chain CDR3 of pt.1-ex08TCR

[0273] Sequence No. 5: Amino Acid Sequence of β-chain CDR3 of pt.1-ex15TCR

[0274] Sequence No. 6: Amino Acid Sequence of β-chain CDR3 of pt.2-ex09TCR Sequence No. 7: Amino Acid Sequence of Peptide CT83-S1 Sequence No. 8: Amino Acid Sequence of Peptide CT83-K1 Sequence No. 9: Amino Acid Sequence of Peptide CT83-K2 Sequence No. 10: Amino Acid Sequence of Peptide CT83-K3 Sequence No. 11: Amino Acid Sequence of Peptide IcHi010_MP41 Sequence No. 12: Amino Acid Sequence of Peptide CT83-7

[0275] Sequence No. 13: Amino Acid Sequence of Peptide CT83-8

[0276] Sequence number 14: Amino acid sequence of peptide CT83-9 Sequence number 15: Amino acid sequence of peptide CT83-10 Sequence number 16: Amino acid sequence of peptide CT83-11 Sequence number 17: Amino acid sequence of CDR1 of the β-chain of pt.2-ex08 TCR Sequence number 18: Amino acid sequence of CDR2 of the β-chain of pt.2-ex08 TCR Sequence number 19: Amino acid sequence of CDR1 of the α-chain of pt.2-ex08 TCR Sequence number 20: Amino acid sequence of CDR2 of the α-chain of pt.2-ex08 TCR Sequence number 21: Amino acid sequence of CDR3 of the α-chain of pt.2-ex08 TCR Sequence number 22: Full-length amino acid sequence of the β-chain of pt.2-ex08 TCR Sequence number 23: Full-length amino acid sequence of the α-chain of pt.2-ex08 TCR Sequence number 24: Amino acid sequence of CDR1 of the β-chain of pt.1-ex08 TCR Sequence number 25: Amino acid sequence of CDR2 of the β-chain of pt.1-ex08 TCR Sequence number 26: Amino acid sequence of CDR1 of the α-chain of pt.1-ex08 TCR Sequence number 27: Amino acid sequence of CDR2 of the α-chain of pt.1-ex08 TCR Sequence number 28: Amino acid sequence of CDR3 of the α-chain of pt.1-ex08 TCR Sequence number 29: Full-length amino acid sequence of the β-chain of pt.1-ex08 TCR Sequence number 30: Full-length amino acid sequence of the α-chain of pt.1-ex08 TCR Sequence number 31: Amino acid sequence of CDR1 of the β-chain of pt.1-ex15 TCR Sequence number 32: Amino acid sequence of CDR2 of the β-chain of pt.1-ex15 TCR Sequence number 33: Amino acid sequence of CDR1 of the α-chain of pt.1-ex15 TCR Sequence number 34: Amino acid sequence of CDR2 of the α-chain of pt.1-ex15 TCR Sequence number 35: Amino acid sequence of CDR3 of the α-chain of pt.1-ex15 TCR Sequence number 36: Full-length amino acid sequence of the β-chain of pt.1-ex15 TCR Sequence number 37: Full-length amino acid sequence of the α-chain of pt.1-ex15 TCR Sequence number 38: Amino acid sequence of CDR1 of the β-chain of pt.2-ex09 TCR Sequence number 39: Amino acid sequence of CDR2 of the β-chain of pt.2-ex09 TCR Sequence number 40: Amino acid sequence of CDR1 of the α-chain of pt.2-ex09 TCR Sequence number 41: Amino acid sequence of CDR2 of the α-chain of pt.2-ex09 TCR Sequence number 42: Amino acid sequence of CDR3 of the α-chain of pt.2-ex09 TCR Sequence number 43: Full-length amino acid sequence of the β-chain of pt.2-ex09 TCR Sequence number 44: Full-length amino acid sequence of the α-chain of pt.2-ex09 TCR

Claims

1. A pharmaceutical composition for the combined administration for treating or preventing cancer expressing Kitakyushu Lung Cancer Antigen 1 (KK-LC-1) in a subject, comprising: a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and T cells, wherein the peptide and the T cells have the following combination (1) or (2) according to the human leukocyte antigen (HLA) type of the subject: (1) When the HLA type of the subject is HLA*C14, it is a combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 1, and the T cells expressing a T cell receptor (TCR) having a β chain comprising the complementary determining region 3 (CDR3) of SEQ ID NO: 2; (2) When the HLA type of the subject is HLA*B15, it is a combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 3, and the T cells expressing a TCR having a β chain comprising the CDR3 of SEQ ID NO: 4, 5, or 6.

2. A pharmaceutical composition for the combined administration for treating or preventing cancer expressing KK-LC-1 in a subject, comprising: a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide; and T cells, wherein the peptide and the T cells have the following combination (1) or (2): (1) A combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 1, and the T cells expressing a TCR having a β chain comprising the CDR3 of SEQ ID NO: 2; (2) A combination of the following peptide and the following T cells, the peptide comprising the amino acid sequence of SEQ ID NO: 3, and the T cells expressing a TCR having a β chain comprising the CDR3 of SEQ ID NO: 4, 5, or 6.

3. The pharmaceutical composition according to claim 2, wherein, The HLA type of the subject is HLA*C14 in the combination of (1) and HLA*B15 in the combination of (2).

4. The pharmaceutical composition according to claim 3, wherein, The HLA type of the subject is HLA*C14:02 in the combination of (1) and HLA*B15:01 in the combination of (2).

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein, The cancer is lung cancer, gastric cancer, breast cancer, or liver cancer.

6. A pharmaceutical composition comprising a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, which is administered in combination with T cells for treating or preventing cancer expressing KK-LC-1 in a subject, wherein the peptide is the following (1) or (2): (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1, which is administered in combination with T cells expressing a TCR having a β chain comprising the CDR3 of SEQ ID NO: 2; (2) A peptide comprising the amino acid sequence of SEQ ID NO: 3, which is administered in combination with T cells expressing a TCR having a β chain comprising the CDR3 of SEQ ID NO: 4, 5, or 6.

7. A pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, the pharmaceutical composition comprising a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, wherein the peptide is the following (1) or (2): (1) A peptide comprising the amino acid sequence of SEQ ID NO: 1; (2) A peptide comprising the amino acid sequence of SEQ ID NO:

3.

8. The pharmaceutical composition according to claim 6 or 7, which is a cancer vaccine.

9. The pharmaceutical composition according to any one of claims 6 to 8, wherein, The cancer is lung cancer, gastric cancer, breast cancer or liver cancer.

10. A peptide comprising the amino acid sequence of SEQ ID NO: 1 or 3, a polynucleotide encoding the peptide, or a vector containing the polynucleotide.

11. A pharmaceutical composition comprising T cells administered in combination with a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide for treating or preventing cancer expressing KK-LC-1 in a subject, wherein the T cells are the following (1) or (2), (1) T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2, administered in combination with a peptide comprising the amino acid sequence of SEQ ID NO: 1; (2) T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6, administered in combination with a peptide comprising the amino acid sequence of SEQ ID NO:

3.

12. A pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, the pharmaceutical composition comprising T cells, wherein the T cells are the following (1) or (2), (1) T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2; (2) T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

13. A polynucleotide, or a vector containing the polynucleotide, encoding a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2, 4, 5, or 6.

14. A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising the step of administering in combination to the subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide and T cells, wherein the peptide and the T cells have the following combination of (1) or (2) according to the human leukocyte antigen (HLA) type of the subject, (1) a combination of a peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2; (2) a combination of a peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

15. A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising: the step of determining the HLA type of the subject; and the step of administering in combination a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide and T cells, (1) when the HLA type of the subject is HLA*C14, administering in combination a peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 2, (2) when the HLA type of the subject is HLA*B15, administering in combination a peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR with a β-chain having a CDR3 comprising SEQ ID NO: 4, 5, or 6.

16. A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising the step of administering to the subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, wherein the peptide is the following (1) or (2), (1) a peptide comprising the amino acid sequence of SEQ ID NO: 1; (2) a peptide comprising the amino acid sequence of SEQ ID NO:

3.

17. A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising: a step of determining the HLA type of the subject; and a step of administering to the subject a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, wherein the peptide is one of the following (1) or (2), (1) When the HLA type of the subject is HLA*C14, a peptide comprising the amino acid sequence of SEQ ID NO: 1; (2) When the HLA type of the subject is HLA*B15, a peptide comprising the amino acid sequence of SEQ ID NO:

3.

18. A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising a step of administering T cells to the subject, wherein the T cells are one of the following (1) or (2), (1) T cells expressing a TCR having a β chain comprising a CDR3 of SEQ ID NO: 2; (2) T cells expressing a TCR having a β chain comprising a CDR3 of SEQ ID NO: 4, 5, or 6.

19. A method for treating or preventing cancer expressing KK-LC-1 in a subject, comprising: a step of determining the HLA type of the subject; and a step of administering T cells to the subject, wherein the T cells are one of the following (1) or (2), (1) When the HLA type of the subject is HLA*C14, T cells expressing a TCR having a β chain comprising a CDR3 of SEQ ID NO: 2; (2) When the HLA type of the subject is HLA*B15, T cells expressing a TCR having a β chain comprising a CDR3 of SEQ ID NO: 4, 5, or 6.

20. Use of a peptide, a polynucleotide encoding the peptide, or a vector containing the polynucleotide, or T cells for manufacturing a pharmaceutical composition for treating or preventing cancer expressing KK-LC-1 in a subject, wherein the pharmaceutical composition has a combination of the peptide and the T cells of the following (1) or (2), (1) A combination of a peptide comprising the amino acid sequence of SEQ ID NO: 1 and T cells expressing a T cell receptor (TCR) having a β chain comprising a CDR3 of SEQ ID NO: 2; (2) A combination of a peptide comprising the amino acid sequence of SEQ ID NO: 3 and T cells expressing a TCR having a β chain comprising a CDR3 of SEQ ID NO: 4, 5, or 6.

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