Anti-hK2 chimeric antigen receptor (CAR)

Abstract

To provide better cancer therapy using CAR-T cells. [Solution] Provided are a chimeric antigen receptor (CAR), for example a CAR targeting human kallikrein-2 (hK2), a cell containing the CAR, a vector encoding the CAR, for example a recombinant expression vector, and a nucleic acid molecule encoding the CAR, a method for producing the CAR, a composition, polypeptide, protein, nucleic acid, host cell, cell population, and a method for treating a disorder, for example cancer, using the disclosed CAR.

Description

[Technical Field]

[0001] (Sequence Listing) This application has been electronically submitted in ASCII format, the entire content of which is incorporated by reference into the present specification as a sequence listing. The ASCII copy described above was created on June 30, 2020 , is named JBI6124WOPCT1_SL.txt, and has a size of 562 ,160 bytes.

[0002] (Field of the Invention) The present invention relates to hK2-targeting chimeric antigen receptors comprising a single-chain variable fragment targeting hK2 (chimeric antigen receptors, CAR), and genetically engineered hK2-targeting immune cells that express the CAR. The present invention also provides nucleic acids encoding the CAR, expression vectors , recombinant cells containing said vectors, and compositions comprising genetically engineered immune cells that express hK2-targeting CAR . Methods for producing the CAR and genetically engineered immune cells, and methods of using genetically engineered immune cells to treat conditions including cancer are also provided.

[0003] (Background) Prostate cancer is the second most frequently diagnosed cancer and the sixth leading cause of cancer death among men, accounting for 14% (903,500) of all new cancer cases and 6% of all cancer deaths among men worldwide (258,400). The course from diagnosis of prostate cancer to death varies depending on the extent of the disease, hormonal status, and the presence or absence of detectable metastasis: localized disease, elevated levels of prostate-specific antigen (PSA) without detectable metastasis after radiotherapy or surgery, and non-castration stage or ​Based on clinical metastasis during the castration phase, it is best classified as a series of clinical stages. Surgery, radiation therapy, or a combination of both can be curative for patients with localized diseases. It is possible, but a significant proportion of these patients do not use elevated PSA levels as evidence. The disease is recurrent, and this is particularly true in high-risk groups, where metastasis occurs, i.e., the end of the disease. This can sometimes lead to a transition to the terminal stage of the disease.

[0004] Androgen depletion therapy (ADT) is the standard treatment, and the generally predictable outcome is P A decrease in SA, a stable phase in which the tumor does not grow, followed by an increase in PSA, and then castration-resistant disease. It is a regrowth. For many years, ADT has been the standard treatment for patients with metastatic prostate cancer. there were.

[0005] Kallikrein-related peptidase 2 (hK2, HK2) is the androgen receptor (AR). It is a trypsin-like enzyme that is driven by and specifically expressed in prostate tissue and prostate cancer. 2 is activated by the transmembrane protease, serine 2 (TMPRSS2), and in the prostate It is secreted into the ducts, where it cleaves semenogerin, an extracellular matrix in the ejaculate fluid, and then the sperm It initiates a cascade that enhances the motility of the cells. hK2 expression is present in prostate and prostate cancer tissue. Although limited, breast cancer after the AR pathway has been appropriately activated by steroid hormones has recently been observed. It has been proven that hK2 is detectable in strains and primary patient samples (U.S. Patent Application Publication). (Case No. 2018 / 0326102). Similar to PSA, when the prostate gland is enlarged or undergoes malignant transformation... When highly structured organization is impaired, catalytically inactive hK2 can be released into the blood. Sexual release occurs.

[0006] In T-cell therapy, genetically modified cells are used to enhance specificity against specific tumor-associated antigens. Isolated T cells are used. Genetic modification involves chimeric antigen receptors (CARs) or exogenous T cells. It can provide new antigen specificity on T cells through the expression of the receptor. The resulting T cells (CAR-T cells) can induce tumor immune responses. There is a need for better cancer therapies that utilize T cells.

[0007] (overview) Chimeric antigen receptors (CARs), such as human kallikrein-2 (hK2), are targeted by C AR, cells containing the CAR, vector encoding the CAR, for example recombinant Current vector, nucleic acid molecule encoding the CAR, method for producing the CAR, and assembly Products, polypeptides, proteins, nucleic acids, host cells, cell populations, and disclosed CARs Methods for treating disorders, such as cancer, using are disclosed herein.

[0008] In one embodiment, a chimeric antigen receptor (CAR) is provided, comprising the following: (a) Extracellular scFv containing scFv that specifically binds to the human kallikrein-2 (hK2) antigen Domain and (b) Transmembrane domain and (c) Intracellular signaling domains optionally containing at least one co-stimulatory domain .

[0009] In some embodiments, CAR is (d) CD8a-Hinge Region It further includes, The transmembrane domain contains a CD8a transmembrane region (CD8A-TM) polypeptide. The intracellular signaling domain in question is a TNF receptor superfamily member 9(C) (D137) component-containing costimulatory domain and T cell surface glycoprotein CD3 zeta chain (CD3) It contains a primary signaling domain that includes the 3z component.

[0010] In some embodiments, the CD8a-hinge region is represented in Sequence IDs 25, 269, 270. Containing an amino acid sequence that is at least 90% identical to 271 or 272, The transmembrane domain contains an amino acid sequence that is at least 90% identical to that of SEQ ID NO: 26. and / or The intracellular signaling domain is at least 90% identical to that of Sequence ID No. 27. A co-stimulatory domain having a sequence, and amino acids that are at least 90% identical to SEQ ID NO: 28. It contains a primary signaling domain having a sequence.

[0011] In another embodiment, the extracellular antigen-binding domain, the transmembrane domain, and the intracellular signaling domain A chimeric antigen receptor (CAR) including the main, wherein the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 4, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, SEQ ID NO: A heavy chain CDR2 having a 315-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 311. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 309, SEQ ID NO: A heavy chain CDR2 having a 310-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 311. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having the amino acid sequence of 3, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 86, SEQ ID NO: 8 Heavy chain CDR2 having the amino acid sequence of 7, and heavy chain having the amino acid sequence of SEQ ID NO: 88 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 92, SEQ ID NO: 9 Heavy chain CDR2 having amino acid sequence 3, and heavy chain having amino acid sequence of SEQ ID NO: 94 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: A heavy chain CDR2 having the amino acid sequence of 103, and having the amino acid sequence of SEQ ID NO: 104 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: A heavy chain CDR2 having a sequence of 106 amino acids, and having the amino acid sequence of SEQ ID NO: 104 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: A heavy chain CDR2 having a sequence of 108 amino acids, and having the amino acid sequence of SEQ ID NO: 109 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 110, SEQ ID NO: A heavy chain CDR2 having an amino acid sequence of 111, and having the amino acid sequence of SEQ ID NO: 109 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: A heavy chain CDR2 having a 112-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 113. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: A heavy chain CDR2 having a sequence of 114 amino acids, and having the amino acid sequence of SEQ ID NO: 113. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: A heavy chain CDR2 having a sequence of 116 amino acids, and having the amino acid sequence of SEQ ID NO: 117 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: A heavy chain CDR2 having a sequence of 119 amino acids, and having the amino acid sequence of SEQ ID NO: 117 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: A heavy chain CDR2 having a 121-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 122. heavy chain CDR3; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 123, SEQ ID NO: A heavy chain CDR2 having a 124-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 122. heavy chain CDR3, Includes, A CAR is provided in which the extracellular antigen-binding domain binds to the hK2 antigen.

[0012] In another embodiment, the extracellular antigen-binding domain, the transmembrane domain, and the intracellular signaling domain A chimeric antigen receptor (CAR) including the main, wherein the extracellular antigen-binding domain is Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 6 Light chain CDR2 having the amino acid sequence of 9, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, SEQ ID NO: 7 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 95, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence of 6, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 312, SEQ ID NO: A light chain CDR2 having a 313-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 91 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 1 A light chain CDR2 having a 26-amino acid sequence, and a light chain having the amino acid sequence of SEQ ID NO: 71. chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 128, SEQ ID NO: A light chain CDR2 having a 129-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 130. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 125, SEQ ID NO: A light chain CDR2 having a 131-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 132. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 133, SEQ ID NO: A light chain CDR2 having a 134-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 135. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 136, SEQ ID NO: A light chain CDR2 having a sequence of 137 amino acids, and having the amino acid sequence of SEQ ID NO: 138. Light chain CDR3; or Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 97, SEQ ID NO: 1 A light chain CDR2 having a 27-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 139 Light chain CDR3, Includes, A CAR is provided in which the extracellular antigen-binding domain binds to the hK2 antigen.

[0013] In some embodiments, the extracellular antigen-binding domain is Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 6 Light chain CDR2 having the amino acid sequence of 9, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; or Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, SEQ ID NO: 7 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 95, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence of 6, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 312, SEQ ID NO: A light chain CDR2 having a 313-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 91 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 1 A light chain CDR2 having a 26-amino acid sequence, and a light chain having the amino acid sequence of SEQ ID NO: 71. chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 128, SEQ ID NO: A light chain CDR2 having a 129-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 130. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 125, SEQ ID NO: A light chain CDR2 having a 131-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 132. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 133, SEQ ID NO: A light chain CDR2 having a 134-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 135. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 136, SEQ ID NO: A light chain CDR2 having a sequence of 137 amino acids, and having the amino acid sequence of SEQ ID NO: 138. Light chain CDR3; or Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 97, SEQ ID NO: 1 A light chain CDR2 having a 27-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 139 Light chain CDR3, It also includes.

[0014] In another embodiment, the extracellular antigen-binding domain, the transmembrane domain, and the intracellular signaling domain A chimeric antigen receptor (CAR) including the main, wherein the extracellular antigen-binding domain is Sequence numbers 1, 2, 3, 74, 75, 147, 148, 149, 150, 151, 152 , and a light chain variable domain (VL) comprising an amino acid sequence selected from 318, and / or teeth Sequence numbers 4, 5, 6, 76, 77, 140, 141, 142, 143, 144, 145 A heavy chain variable domain (VH) containing an amino acid sequence selected from 146 and 317, Includes, The extracellular antigen-binding domain binds to the human kallikrein-2 (hK2) antigen. AR will be provided.

[0015] In some embodiments, the extracellular antigen-binding domain is Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; The light chain variable region includes the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of SEQ ID NO: 76. Heavy chain variable region; The light chain variable region includes the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 77. Heavy chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 147 and amino acid sequence of SEQ ID NO: 140 Heavy chain variable region including; The light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 141 Includes heavy chain variable region; The light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 140 Includes heavy chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 148 and amino acid sequence of SEQ ID NO: 142 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 149 and amino acid sequence of SEQ ID NO: 143 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 150 and amino acid sequence of SEQ ID NO: 144 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 151 and amino acid sequence of SEQ ID NO: 145 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 152 and amino acid sequence of SEQ ID NO: 146 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 318 and amino acid sequence of SEQ ID NO: 317 Heavy chain variable region including, Includes.

[0016] In some embodiments, the extracellular antigen-binding domain uses a single-stranded variable fragment (scFv) Includes. In some embodiments, scFv has a light chain variable region (VL) and a heavy chain variable region (V A linker polypeptide is included between H and H. In some embodiments, the linker polypeptide The cido contains the amino acid sequence of SEQ ID NO: 7. In some embodiments, scFv is the sequence Amino acids selected from the groups consisting of numbers 8-23, 169-184, and 340-343. Includes a sequence. In some embodiments, the extracellular antigen-binding domain is a signaling polypeptide. It contains. In some embodiments, the signal polypeptide is the amino acid of SEQ ID NO: 24. Includes arrays.

[0017] In some embodiments, the intracellular signaling domain is a TNF receptor superfa Millie member 9 (CD137) component, T cell surface glycoprotein CD3 zeta chain (CD3 z) Components, surface antigen classification (CD27) components, surface antigen classification superfamily member components It contains polypeptide components selected from the group consisting of minutes and combinations thereof. In that embodiment, component CD137 includes the amino acid sequence of SEQ ID NO: 27. In the embodiment, the CD3z component includes the amino acid sequence of SEQ ID NO: 28. Several embodiments In this state, the intracellular signaling domain contains the amino acid sequence of SEQ ID NO: 45. In this embodiment, the transmembrane domain is a CD8a transmembrane region (CD8A-TM) polypeptide It includes. In some embodiments, the CD8a-TM polypeptide is the A of SEQ ID NO: 26. Contains a mino acid sequence.

[0018] In various embodiments, the CAR ligates its transmembrane domain to an extracellular antigen-binding domain. Further includes a hinge region. In some embodiments, the hinge region is the CD8a hinge region. Yes. In some embodiments, the CD8a-hinge region is represented by sequence numbers 25, 269, and 27. The amino acid sequence comprises 0, 271, or 272. In some embodiments, extracellular antigen clusters The combined domains are sequence numbers 29-44, 78-81, 221-236, and 273-308. It comprises an amino acid sequence selected from the group consisting of the following. In some embodiments, the CAR is a Amino acids selected from the groups consisting of row numbers 46-61, 82-85, and 205-220. Includes arrays.

[0019] In another embodiment, isolated lymphocytes expressing any of the above CARs are provided. In this embodiment, the lymphocytes are T lymphocytes. In some embodiments, the lymphocytes are These are natural killer (NK) cells.

[0020] Also provided are isolated nucleic acid molecules encoding any of the above CARs. A vector containing the nucleic acid molecule is also provided. Furthermore, cells expressing nucleic acid molecules are also provided.

[0021] Furthermore, a pharmaceutical comprising an effective amount of any of the above lymphocytes and a pharmaceutically acceptable excipient, Compositions are also provided.

[0022] In one embodiment, the present disclosure provides a CAR according to the present disclosure for use in a therapeutic method.

[0023] In one embodiment, the present disclosure provides lymphocytes according to the present disclosure for use in a therapeutic method.

[0024] In one embodiment, the present disclosure relates to a composition according to the present disclosure for use in a therapeutic method, for example, a pharmaceutical A composition is provided.

[0025] In one aspect, the Disclosure provides a CAR according to the Disclosure for use in a method of treating cancer. do.

[0026] In one aspect, the Disclosure provides lymphocytes according to the Disclosure for use in a method of treating cancer. To provide.

[0027] In one embodiment, the present disclosure relates to a composition for use in a method of treating cancer, for example The present invention provides a pharmaceutical composition.

[0028] In one embodiment, the cancer is prostate cancer.

[0029] In one embodiment, the cancer is androgen receptor (AR) expressing breast cancer.

[0030] In another embodiment, a method is provided for treating a subject having cancer. The method involves a therapeutically effective dose of Administer one of the lymphocytes to a subject in need, thereby... The spheres induce the killing of cancer cells in the target. In some embodiments, The cancer is prostate cancer. In some embodiments, the cancer is androgen receptor (AR) I currently have breast cancer.

[0031] In another embodiment, a method for targeting and killing cancer cells, wherein the cancer cells are subjected to the lymphocytes This includes bringing the lymphocytes into contact with the cancer cells, thereby inducing the killing of the cancer cells. A method is provided. In some embodiments, the cancer cells are prostate cancer cells. In that embodiment, the cancer is androgen receptor (AR) expressing breast cancer.

[0032] In another embodiment, a method for detecting the presence of cancer in a subject, (a) The cell sample obtained from the subject is brought into contact with one of the above CARs, and Furthermore, to form a CAR-cell complex, (b) detecting the complex, and hereby, the detection of the complex in the subject Indicating the presence of cancer, A method is provided that includes this. [Brief explanation of the drawing]

[0033] The foregoing will become clear from a more detailed description of the exemplary embodiments shown in the accompanying drawings below. It will probably happen.

[0034] This patent application document includes at least one color drawing. Copies of this patent application may be made by the Japan Patent Office upon request and payment of the required fees. It will be provided. [Figure 1A] This shows the construction and expression of hK2 CAR in human T cells. Figure 1A shows a schematic diagram of the construct for hK2 CAR. TM: Transmembrane. [Figure 1B] This paper describes the construction and expression of the hK2 CAR in human T cells. Figure 1B shows hK2 CAR expression on the T cell surface. Primary human T cells were electroporated with either no mRNA (mock) or 10 μg of mRNA expressing either the hK2 scFv CAR or an unrelated control CAR. 24 hours after electroporation, the cells were stained with 2 μg / mL biotinylated L protein and streptavidin-conjugated PE (Figure 1B), and CAR surface expression was measured by flow cytometry. [Figure 1C]This paper describes the construction and expression of the hK2 CAR in human T cells. Figure 1C shows hK2 CAR expression on the T cell surface. Primary human T cells were electroporated with either no mRNA (mock) or 10 μg of mRNA expressing either the hK2 scFv CAR or an unrelated control CAR. 24 hours after electroporation, the cells were stained with biotinylated hK2 (1 μg / mL) and streptavidin-conjugated PE (Figure 1C), and CAR surface expression was measured by flow cytometry. [Figure 2] This study demonstrates the cytotoxicity of hK2-expressing tumor cells by CAR-T cells in a 20-hour flow-based assay at a specified effector-to-target cell (E / T) ratio. Target cells (Vcap and DU145) were labeled with Cell Trace Violet (CTV) fluorescent dye 24 hours after transient transfection and then co-cultured with hK2 CAR-T cells. Mock T cells served as a negative effector control. The mortality (killing) percentage is the ratio of the absolute number of viable (viable dye-negative) target (CTV-positive) cells remaining in the co-culture to the number of viable targets cultured without CAR-T cells. [Figure 3A]This exhibits real-time hK2-specific CAR-T cell-mediated cytotoxicity. Normalized cell index (CI) plots of target cells (Figure 3A) VCaps(5E4) incubated with mock, 10 μg mRNA electroporated (24 hours after transfection) hK2 11B6 CAR LH, or control CAR-T cells for approximately 72 hours at different E / T ratios. When seeded alone, target cells adhere to the plate, proliferate, and increase CI readout. Addition of T cells to target cells, both hK2 CAR- and control CAR-T cells, induced cytolysis of hK2(+) VCaps cells and a subsequent progressive decrease in CI at E / T ratios ranging from 5:1 to 0.156:1. The decrease in CI values ​​after the addition of effector cells reflects the loss of viability of target cells. As shown in Figure 3B, parallel experiments were also conducted using hK2(-)DU145 target cells; in the E:T ratio tested in this experiment, hK2 and control CAR T cells did not decrease CI after addition and did not show cell-lytic activity. The Y-axis represents the normalized CI, created and displayed in real time by RTCA software. The X-axis represents the cell culture time and processing time (hours). The mean CI is plotted with ± standard deviation. [Figure 3B]This exhibits real-time hK2-specific CAR-T cell-mediated cytotoxicity. Normalized cell index (CI) plots of target cells (Figure 3B) DU145(5E3) incubated with mock, 10 μg mRNA electroporated (24 hours after transfection) hK2 11B6 CAR LH, or control CAR-T cells for approximately 72 hours at different E / T ratios. When seeded alone, target cells adhere to the plate, proliferate, and increase CI readout. Addition of T cells to target cells, both hK2 CAR- and control CAR-T cells, induced cytolysis of hK2(+) VCap cells and a subsequent progressive decrease in CI at E / T ratios ranging from 5:1 to 0.156:1. The decrease in CI values ​​after the addition of effector cells reflects the loss of viability of target cells. As shown in Figure 3B, parallel experiments were also conducted using hK2(-)DU145 target cells; in the E:T ratio tested in this experiment, hK2 and control CAR T cells did not decrease CI after addition and did not show cell-lytic activity. The Y-axis represents the normalized CI, created and displayed in real time by RTCA software. The X-axis represents the cell culture time and processing time (hours). The mean CI is plotted with ± standard deviation. [Figure 4] This shows the production of Th1 cytokine interferon-γ (IFN-γ) by antigen-stimulated CAR-T cells. Supernatants were collected from xCELligence-based killing assays (see Figures 3A-3B) after approximately 70 hours of co-culture (VCap#5E4, DU145#5E3). hK2 CAR LH and control CAR-modified T cells secrete IFN-γ during co-culture with hK2-expressing VCap cells, but not during co-culture with hK2-negative DU145 cells. Mean IFN-γ concentration ± standard deviation (pg / mL) from duplicate cultures is shown. [Figure 5A]This shows the evaluation of the antigen-dependent activity of the 11B6 heat-stabilized scFv CAR clone using a JNL reporter assay. Figure 5A shows the transduction of various hK2 CAR constructs into Jurkat cells (called JNL cells) containing a luciferase gene driven by a signal transduction-responsive NFAT promoter. Expression was determined using biotinylated hK2, followed by streptavidin-conjugated PE. [Figure 5B] This report shows the evaluation of the 11B6 heat-stabilized scFv CAR clone for antigen-dependent activity using a JNL reporter assay. Figure 5B shows that luciferase expression occurred in JNL cells upon binding of the CAR construct to its allogeneic cell antigen (hK2 on target cells). JNL cells containing the specified CAR clone and untransduced JNL cells (UTD) were co-cultured with target cell lines (VCap or DU145 cells), and luciferase activity was measured as luminescence intensity. If the luminescence intensity was 1.5 times higher than that of UTD cells in the presence of antigen-expressing cells, the clone was considered active. [Figure 6] hK2 CAR expression on the T cell surface is shown. Primary human T cells were transduced with 11B6 heat-stabilized and parental scFv-based CAR lentivirus (multiple infections (MOI): 3). Fourteen days after transduction, CAR expression was determined by biotinylated hK2 (1 μg / mL), followed by streptavidin-conjugated PE. [Figure 7A] This report describes the evaluation of 11B6 thermostabilized scFv CAR-T cells for antigen-dependent cytotoxicity using a real-time IncuCyte killing assay. Thermostabilized hK2 CAR-T cells and parental 11B6 CAR-T cells were co-incubated with VCaP cells (Figure 7A) for 88 hours. The effector-to-target (E / T) ratio was calculated based on CAR expression data (see Figure 6). Target cells stably expressed red nuclear dye, as measured in real time by the IncuCyte imaging system. Tumor cell growth inhibition (%) = (initial number of surviving target cells - current number of surviving target cells) / initial number of surviving cells × 100 (%). [Figure 7B]This report describes the evaluation of 11B6 heat-stabilized scFv CAR-T cells for antigen-dependent cytotoxicity using a real-time IncuCyte killing assay. Heat-stabilized hK2 CAR-T cells and parental 11B6 CAR-T cells were co-incubated with PC3 cells (Figure 7B) for 88 hours. The effector-to-target (E / T) ratio was calculated based on CAR expression data (see Figure 6). Target cells stably expressed red nuclear dye, as measured in real time by the IncuCyte imaging system. Tumor cell growth inhibition (%) = (initial number of surviving target cells - current number of surviving target cells) / initial number of surviving cells × 100 (%). [Figure 7C] This shows the percentage of tumor cell growth inhibition in hK2-positive VCaP cells at effector:target ratios of 1:1 or 0.5:1 by T cells transduced with CAR1-10 or parental 11B6_HL or 11B6_LH in a real-time incucyte killing assay to evaluate antigen-dependent cytotoxicity. Tumor cell growth inhibition (%) = (Initial number of surviving target cells - Current number of surviving target cells) / Initial number of surviving cells × 100 (%). [Figure 7D] This shows the percentage of tumor cell growth inhibition in PC3 cells at an effector:target ratio of 1:1 by T cells transduced with CAR1-10 or parental 11B6_HL or 11B6_LH in a real-time incucyte killing assay to evaluate antigen-dependent cytotoxicity. Tumor cell growth inhibition (%) = (Initial number of surviving target cells - Current number of surviving target cells) / Initial number of surviving cells × 100 (%). [Figure 7E] The figures show the percentage of tumor cell growth inhibition of hK2-positive VCaP cells by transduced CAR-T cells using CAR17 (B413HL in the figure), CAR18 (B413LH in the figure), CAR19 (B359HL in the figure), and CAR20 (B359LH in the figure) in a real-time incucyte killing assay to evaluate antigen-dependent cytotoxicity. Tumor cell growth inhibition (%) = (Initial number of surviving target cells - Current number of surviving target cells) / Initial number of surviving cells × 100 (%). [Figure 7F]The figures show the percentage of tumor cell growth inhibition in hK2-negative DU145 cells by transduced CAR-T cells of CAR17 (B413HL in the figure), CAR18 (B413LH in the figure), CAR19 (B359HL in the figure), and CAR20 (B359LH in the figure) in a real-time incucyte killing assay to evaluate antigen-dependent cytotoxicity. Tumor cell growth inhibition (%) = (Initial number of surviving target cells - Current number of surviving target cells) / Initial number of surviving cells × 100 (%). [Figure 8] This shows cytokine release by hK2 lenti-CAR-T cells. The supernatant collected from co-cultures of hK2 CAR-T cells with VCap cells overnight (approximately 20 hours) at a 1:1 E / T ratio was analyzed using the 13-plex Milliplex Human High Sensitivity T Cell Kit (HSTCMAG28SPMX13). hK2 CAR-modified T cells secrete cytokines during co-culture with hK2-expressing VCap cells, but the amount is minimal in untransduced T cells (UTDs). The mean cytokine concentration ± standard deviation (pg / mL) from the duplicated culture is shown. [Figure 9] This study demonstrates IFN-γ release by hK2 lenti-CAR-T cells. Supernatants were collected from hK2 CAR-T cells co-cultured overnight (approximately 20 hours) with VCaP and DU145 (5E4 cells) cells in a 1:1 E / T ratio. hK2 CAR-modified T cells secreted IFN-γ during co-culture with hK2-expressing VCaP cells but not during co-culture with hK2-negative DU145 cells. The mean IFN-γ concentration ± standard deviation (pg / mL) from the duplicated cultures is shown. CD3 / 28 bead-stimulated T cells and T cells alone were used as positive and negative controls, respectively. [Figure 10A]The results obtained from the hK2 CAR-T cell proliferation assay are shown. (Figures 10A, 10C) hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with hK2(+) VCaps and hK2(-) ​​DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. (Figures 10B, 10D) CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. The data in Figure 10E show that hK2 CAR-T cells proliferate more robustly than CD3 / 28 bead-positive controls after co-culture with VCap cells for 5 days. T cells genetically engineered with different CAR constructs exhibited different proliferative activity and different CAR+ T cell counts. CAR+ T cell counts were based on the mean absolute cell count + / - SEM from three technical replicas. [Figure 10B]The results obtained from the hK2 CAR-T cell proliferation assay are shown. (Figures 10A, 10C) hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with hK2(+) VCaps and hK2(-) ​​DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. (Figures 10B, 10D) CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. The data in Figure 10E show that hK2 CAR-T cells proliferate more robustly than CD3 / 28 bead-positive controls after co-culture with VCap cells for 5 days. T cells genetically engineered with different CAR constructs exhibited different proliferative activity and different CAR+ T cell counts. CAR+ T cell counts were based on the mean absolute cell count + / - SEM from three technical replicas. [Figure 10C]The results obtained from the hK2 CAR-T cell proliferation assay are shown. (Figures 10A, 10C) hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with hK2(+) VCaps and hK2(-) ​​DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. (Figures 10B, 10D) CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. The data in Figure 10E show that hK2 CAR-T cells proliferate more robustly than CD3 / 28 bead-positive controls after co-culture with VCap cells for 5 days. T cells genetically engineered with different CAR constructs exhibited different proliferative activity and different CAR+ T cell counts. CAR+ T cell counts were based on the mean absolute cell count + / - SEM from three technical replicas. [Figure 10D]The results obtained from the hK2 CAR-T cell proliferation assay are shown. (Figures 10A, 10C) hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with hK2(+) VCaps and hK2(-) ​​DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. (Figures 10B, 10D) CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. The data in Figure 10E show that hK2 CAR-T cells proliferate more robustly than CD3 / 28 bead-positive controls after co-culture with VCap cells for 5 days. T cells genetically engineered with different CAR constructs exhibited different proliferative activity and different CAR+ T cell counts. CAR+ T cell counts were based on the mean absolute cell count + / - SEM from three technical replicas. [Figure 10E]The results obtained from the hK2 CAR-T cell proliferation assay are shown. (Figures 10A, 10C) hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with hK2(+) VCaps and hK2(-) ​​DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. (Figures 10B, 10D) CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. The data in Figure 10E show that hK2 CAR-T cells proliferate more robustly than CD3 / 28 bead-positive controls after co-culture with VCap cells for 5 days. T cells genetically engineered with different CAR constructs exhibited different proliferative activity and different CAR+ T cell counts. CAR+ T cell counts were based on the mean absolute cell count + / - SEM from three technical replicas. [Figure 11A] The results of an assay to determine the expression level of hK2 CAR in primary T cells using biotin-labeled hK2 protein are shown. Figure 11A is a schematic diagram of the construct for hK2 CAR. TM: Transmembrane. [Figure 11B]The results of an assay to determine the expression level of hK2 CAR in primary T cells using biotin-labeled hK2 protein are shown. In Figure 11B, primary human T cells were transduced with novel hK2 scFv-based KL2B413 HL and LH, as well as re-humanized 11B6 KL2B359 HL and LH CARS lentiviruses (MOI 3). Fourteen days after transduction, CAR expression was determined by biotinylated hK2 (1 μg / mL), followed by streptavidin-conjugated PE. Figure 11B provides an overview of the percentage (positive %) of hK2 CAR+ T cells detected by the analyzed novel KL2B413 HL and LH, and re-humanized 11B6 KL2B359 HL and LH CARS. As shown, different clones have different CAR expression levels ranging from 45.1% to 59.9%. All CAR T+ cells were normalized to equal CAR+ T cells for subsequent functional assays. [Figure 12A] The results of the evaluation of novel hK2 KL2B413 and re-humanized 11B6 CAR clones in the JNL reporter assay for antigen-dependent activity are shown. In Figure 12A, various hK2 CAR constructs were transduced into Jurkat cells (called JNL cells) containing a luciferase gene driven by a signal transduction-responsive NFAT promoter. Expression was determined by biotinylated hK2 followed by streptavidin-conjugated PE. CAR expression in transduced JNL cells was confirmed for selected clones, with re-humanized 11B6 KL2B359 CAR expressed in 46-50% of cells and KL2B413 CAR expressed in 73.7-96% of cells. [Figure 12B]This report presents the results of the evaluation of novel hK2 KL2B413 and re-humanized 11B6 CAR clones in a JNL reporter assay for antigen-dependent activity. The data shown in Figure 12B demonstrate that luciferase expression in JNL cells occurs upon binding of the hK2 CAR construct to its allogeneic antigen (hK2 on target cells). JNL cells containing the specified CAR clones and JNL cells (UTD) were co-cultured with target cell lines (VCap, LNCap / Hk2, LNCap, C4-2B, 22Rv1, or DU145 cells), and luciferase activity was measured as luminescence intensity. A clone was considered active if its luminescence intensity was 1.5 times higher than that of UTD cells in the presence of antigen-expressing cells. Antigen-dependent activation was not observed for the novel KL2B413 or the re-humanized 11B6 HL and LH CARs. [Figure 13A] This report presents the results of evaluating HK2 CAR-T cells in a real-time IncuCyte killing assay for antigen-dependent cytotoxicity. HK2 CAR-T cells were co-incubated with HK2+VCaP cells (Figure 13A) for 96 hours. The effector-to-target (ET) ratio was calculated based on CAR expression data (Figure 11B). Target cells stably expressed red nuclear dye, as measured in real time by the IncuCyte imaging system. Tumor cell growth inhibition (%) = (Initial number of surviving target cells - Current number of surviving target cells) / Initial number of surviving cells × 100 (%). [Figure 13B] This report presents the results of evaluating HK2 CAR-T cells in a real-time IncuCyte killing assay for antigen-dependent cytotoxicity. HK2 CAR-T cells were co-incubated with HK2-DU145 cells (Figure 13B) for 96 hours. The effector-to-target (ET) ratio was calculated based on CAR expression data (Figure 11B). Target cells stably expressed red nuclear dye, as measured in real time by the IncuCyte imaging system. Tumor cell growth inhibition (%) = (initial number of surviving target cells - current number of surviving target cells) / initial number of surviving cells × 100 (%). [Figure 14]The data shows the production of the Th1 cytokine IFN-gamma by antigen-stimulated CAR-T cells. IFN-γ produced by cytotoxic T cells is crucial for the immune surveillance function of tumors, directly inhibiting proliferation and inducing apoptosis in some malignant tumors in vivo and in vitro. To determine whether hK2 CAR-modified human T cells can be recognized and activated by hK2(+) tumor cells, primary T cells containing specified CAR clones and control untransduced T cells (UTDs) were co-cultured with target cell lines (LNCap / Hk2, LNCap, C4-2B, 22Rv1, or DU145 cells), and the supernatant was collected for IFN-g concentration measurement. As illustrated, hK2 CAR-modified T cells secrete IFN-gamma during co-culture with hK2-expressing LNCap / hK2 cells, even in C4-2B and LNCap cells with very low HK2 expression, but not in hK2-negative DU145 cells. The undisclosed control CAR secreted much larger amounts of IFN-γ due to its significantly higher antigen expression level than hK2. Mean IFN-γ concentrations ± SD (pg / mL) from duplicate cultures are shown. [Figure 15]The results of a CD107a degranulation assay to evaluate the antitumor activity of hK2 CAR T cells are shown. Target cells (5 × 10⁴ cells) were co-cultured with an equal number of effector cells at 0.1 mL / well in a 96-well plate. The control well contained only T cells. Anti-CD107a (5 μL / well) was added in addition to 1 μL / sample of monensin (BD Biosciences) and incubated at 37°C for 4 hours. Cells were washed twice with PBS, stained for hK2 CAR, CD3, and CD8 expression, and analyzed using a BD Fortessa flow cytometer. As shown, the presence of hK2 antigen on Vcap cells increased CD107A+ hK2 CAR(+) T cells as measured by flow cytometry, but not in hK2 CAR(-) and UTD T cells, while CAR T cells alone without tumor cell stimulation showed only background staining (<2%). Forbol 12-myristate 13-acetate (PMA) combined with ionomycin (eBioscience® cell stimulation cocktail; 500×) was a consistent inducer of CD107a cell surface expression in this 4-hour cell culture model and was used as a positive control. Results are shown for representative donor T cells, and these results are reproducible in multiple donors. [Figure 16A]The results of the hK2 CAR T cell proliferation assay are shown. hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with HK2(+)VCap and HK2(-)DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. Gating with CD3+ T cells promoted the proliferation of T cells genetically engineered with all CAR constructs, as shown in Figures 16A–16B, and the upregulation of the activation marker CD25, as shown in Figures 16C–16D, but not with hK2(-)DU145 cells. CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. T cells only without any stimulation did not proliferate, while CD3 / 28 bead-stimulated T cells showed equivalent proliferation patterns. As shown, hK2 CAR+ T cells proliferated more robustly than CD3 / 28 bead-positive controls after co-culture with VCaps cells for 5 days. T cells genetically engineered with different CAR constructs had different proliferative activity and showed different CAR+ T cell counts. Figures 16B and 16D summarize the percentages of proliferating T cells and CD25-expressing T cells based on mean absolute cell number + / - SEM from duplicates. [Figure 16B]The results of the hK2 CAR T cell proliferation assay are shown. hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with HK2(+)VCap and HK2(-)DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. Gating with CD3+ T cells promoted the proliferation of T cells genetically engineered with all CAR constructs, as shown in Figures 16A–16B, and the upregulation of the activation marker CD25, as shown in Figures 16C–16D, but not with hK2(-)DU145 cells. CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. T cells only without any stimulation did not proliferate, while CD3 / 28 bead-stimulated T cells showed equivalent proliferation patterns. As shown, hK2 CAR+ T cells proliferated more robustly than CD3 / 28 bead-positive controls after co-culture with VCaps cells for 5 days. T cells genetically engineered with different CAR constructs had different proliferative activity and showed different CAR+ T cell counts. Figures 16B and 16D summarize the percentages of proliferating T cells and CD25-expressing T cells based on mean absolute cell number + / - SEM from duplicates. [Figure 16C]The results of the hK2 CAR T cell proliferation assay are shown. hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with HK2(+)VCap and HK2(-)DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. Gating with CD3+ T cells promoted the proliferation of T cells genetically engineered with all CAR constructs, as shown in Figures 16A–16B, and the upregulation of the activation marker CD25, as shown in Figures 16C–16D, but not with hK2(-)DU145 cells. CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. T cells only without any stimulation did not proliferate, while CD3 / 28 bead-stimulated T cells showed equivalent proliferation patterns. As shown, hK2 CAR+ T cells proliferated more robustly than CD3 / 28 bead-positive controls after co-culture with VCaps cells for 5 days. T cells genetically engineered with different CAR constructs had different proliferative activity and showed different CAR+ T cell counts. Figures 16B and 16D summarize the percentages of proliferating T cells and CD25-expressing T cells based on mean absolute cell number + / - SEM from duplicates. [Figure 16D]The results of the hK2 CAR T cell proliferation assay are shown. hK2 CAR and untransduced (UTD) T cells were labeled with CellTrace Violet (CTV, 5 μM) and co-cultured with HK2(+)VCap and HK2(-)DU145 cells. After 5 days of co-culture, cells were collected and stained with CD3, CD25, NearIR live / dead dye, and hK2CAR. Flow cytometry analysis was performed using a Fortessa flow cytometer with Flowjo software. Lymphocytes were identified by live CD3, and the frequency of CAR+ T cells with CTV dye dilution and the activation marker CD25 was determined. Gating with CD3+ T cells promoted the proliferation of T cells genetically engineered with all CAR constructs, as shown in Figures 16A–16B, and the upregulation of the activation marker CD25, as shown in Figures 16C–16D, but not with hK2(-)DU145 cells. CD3 / 28 bead-stimulated T cells and T cells only were used as positive and negative controls, respectively. T cells only without any stimulation did not proliferate, while CD3 / 28 bead-stimulated T cells showed equivalent proliferation patterns. As shown, hK2 CAR+ T cells proliferated more robustly than CD3 / 28 bead-positive controls after co-culture with VCaps cells for 5 days. T cells genetically engineered with different CAR constructs had different proliferative activity and showed different CAR+ T cell counts. Figures 16B and 16D summarize the percentages of proliferating T cells and CD25-expressing T cells based on mean absolute cell number + / - SEM from duplicates. [Figure 17A] The sequence alignments of the VH domains of mu11B6, hu11B6, KL2B357, KL2B358, KL2B359, KL2B360, HCF3, and HCG5 are shown. Figure 17A discloses sequence numbers 317, 5, 140, 141, 77, 140, 6, 4, and 345, respectively, in order of appearance. [Figure 17B]Protected segments mapped onto the hK2 antigen sequence are shown to visualize the binding epitopes of the hK2 antibody identified via HDX-MS. Figure 17B discloses sequence numbers 351, 351, 351, 351, 351, and 351, respectively, in order of appearance.

[0035] (Detailed description) The following describes exemplary embodiments.

[0036] This disclosure relates to a chimeric antigen receptor (CAR) that targets human kallikrein-2 (hK2). , cells containing such CARs, and cancer (e.g., pre- This invention provides a method for treating prostate cancer or AR-expressing breast cancer.

[0037] The CAR of the present invention has antigen specificity for hK2. The phrase "induces an antigen-specific response" means, as used herein, that CARs have a specific response to an antigen. It binds heterologously and can be recognized immunologically, resulting in the binding of CAR to the hK2 antigen. This means that an immune response is triggered by antigen specificity and the ability to recognize target cells. Methods for testing CAR are known in the art.

[0038] This disclosure also relates to relevant nucleic acids, recombinant expression vectors, host cells, cell populations, antibodies, and The invention also provides a pharmaceutical composition relating to the antigen-binding portion and the CAR of the present invention.

[0039] Some aspects of the present invention are described below with reference to examples provided for illustrative purposes only. To provide a complete understanding of the present invention, numerous specific details, relationships, and methods are described. Please understand that this is the case. However, those skilled in the art will know that the present invention is not a concrete detail of the present invention. It must be possible to carry out the procedure without one or more of the following, or by other methods, protocols, or reagents. It will be readily apparent that this can be done in cell lines and animals. Because actions can occur in different orders and / or simultaneously with other actions or events, the present invention The order of the actions or events described is not limited by the order of the examples. Furthermore, the method according to the present invention Not all of the exemplified actions, steps, or events are necessarily required to implement the method. Many of the techniques and procedures described or referenced herein are of a degree that can be understood by those skilled in the art. It is well understood and commonly used using conventional methods.

[0040] Unless otherwise defined, all technical terms, notations, and other scientific terms used herein , or technical terms, have meanings that are ordinarily understood by those skilled in the art to whom this invention relates. Intention. In some cases, terms that have a commonly understood meaning are used for clarity. and / or as defined herein for immediate reference, including such definitions herein. To do so will always reveal a substantial difference from what is generally understood in the relevant technical field. It should not be interpreted in any way. Terms such as those defined in commonly used dictionaries, As having meanings consistent with those meanings in the context of related technologies, and / or this It is further understood that it needs to be interpreted as being defined otherwise in the specification. It will probably happen.

[0041] When a list is presented, unless otherwise specified, each individual element of that list and its list It should be understood that all combinations of strikes are distinct embodiments. For example, "A, The list of embodiments presented as "B" or "C" is, It is interpreted as including "A or B", "A or C", "B or C", or "A, B, or C". It should be done.

[0042] definition When used in this specification and the attached "Claims", "a", "an", And the singular form "the" indicates plural unless otherwise explicitly indicated. It encompasses the referent. Therefore, for example, the reference "a cell" can refer to two things. This includes combinations of three or more cells, and similar structures.

[0043] Transition phrases: "comprising," "consisting essential" The terms "ly of" and "consisting" are generally accepted in patent terminology. It is intended to imply the meaning of "to be equipped / include (compri "Sing" is synonymous with "include," "contain," or "characterize," and is comprehensive or non-comprehensive. This is restrictive and does not exclude other elements or methods not listed. (ii) "consisting of" is specified in the claims. (iii) "essentially consisting of" This refers to the specified materials or processes, as well as the "basic and novel features" of the claimed invention. The scope of the patent claims is limited to those that do not substantially affect (multiple) the patent. Embodiments described in relation to "comprising" (or equivalent) are also implementations. As a matter of state, I propose to present independently the descriptions of "consisting of" and "essentially becoming from". To provide.

[0044] "Activation," "stimulation," "activated," or "stimulated" refers to the activation marker. This can lead to the expression of - cytokines, cytokine production, or mediation of target cell proliferation or cytotoxicity. This refers to the induction of a change in the biological state of a cell. Cells are activated by a primary stimulus signal. This can happen. Co-stimulatory signals amplify the magnitude of the primary signal and suppress cell death after initial stimulation. This allows for a more durable activation state, and consequently, higher cytotoxicity. This results in "co-stimulatory signals" such as primary signals like TCR / CD3 ligation. In combination with this, the proliferation of T cells and / or NK cells and / or the enhancement of key molecules This refers to a signal that causes preregulation or downregulation.

[0045] An "antigen-binding fragment" or "antigen-binding domain" is a part of a protein that binds to an antigen. It refers to a synthetic polypeptide, an enzymatically available polypeptide, or a genetically modified polypeptide. These may be modified polypeptides, which may include antigen-binding immunoglobulins Some, for example, VH, VL, VH and VL, Fab, Fab', F(ab')2, Fd and Domain antibodies (dAb) consisting of a fragment of Fv, one VH domain, or one VL domain. ), shark variable IgNAR domain, camelidized VH domain, VHH domain, FR3-C A minimal recognition unit consisting of amino acid residues that mimic the CDR of antibodies, such as the DR3-FR4 region. T, HCDR1, HCDR2, and / or HCDR3, as well as LCDR1, LCDR2, and / or LCDR3, alternative scaffolds that bind to the antigen, and antigen binding. Examples include multispecific proteins containing fragments. Antigen-binding fragments (such as VH and VL) are The VH and VL domains are linked to each other via a synthetic linker, allowing for expression as separate single strands. When this occurs, the VH / VL domains pair intramolecularly or intermolecularly to form a monovalent antigen-binding domain. For example, forming a single-chain Fv (scFv) or a diabody. It is possible to form various types of single-chain antibody designs. Antigen-binding fragments are also two To genetically engineer heavy-specific and multispecific proteins, single-specific or multispecific Other possible antibodies, proteins, antigen-binding fragments, or alternative scaffolds Even if conjugated.

[0046] "Cancer" is a widespread disease characterized by the uncontrolled growth of abnormal cells in the body. This refers to a typical group. Uncontrolled cell division and growth result in the form of malignant tumors that invade adjacent tissues. It can cause growth and metastasize to distal parts of the body via the lymphatic system or bloodstream. "Cancer" or " "Cancer tissue" may include tumors.

[0047] A "full-length antibody" consists of two heavy chains (HC) linked together by disulfide bonds. It is composed of two light chains (LCs) and their polymers (e.g., IgM). The chain consists of a heavy chain variable domain (VH) and a heavy chain constant domain, and the heavy chain constant domain It consists of subdomains CH1, hinge, CH2, and CH3. Each light chain is light chain variable. It consists of a domain (VL) and a light chain constant domain (CL). VH and VL are part of the Fray Complementarity determination regions scattered throughout the framework region (FR) It can be further divided into a region of hypervariability called the ity determining region (CDR). H and VL are in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1 , FR2, CDR2, FR3, CDR3, and FR4 are arranged as three CDRs and 4 It consists of two FR segments.

[0048] Human antibodies are optimized to produce the minimum possible immune response when administered to human subjects. This refers to antibodies that have been modified. The variable region of human antibodies is derived from the human immunoglobulin sequence. If the antibody includes a constant region or a part of a constant region, the constant region is also human immunoglobulin Derived from the row. Human antibodies are derived from human germline immunoglobulins or rearranged in the variable region of human antibodies. When obtained from a system using the immunoglobulin gene, the sequence is of human origin. This includes a heavy chain variable region and a light chain variable region. Such exemplary systems are used to phages. A sprayed human immunoglobulin gene library and human immunoglobulin gene loci. This includes transgenic non-human animals possessing "human antibodies," such as mice or rats. " is typically a system used to obtain human antibodies and human immunoglobulin gene loci. The difference lies in the intentional introduction of somatic mutations into the framework or CDR. By means of, or both of these, the amino acid content when compared to immunoglobulins expressed in humans It contains differences. Typically, "human antibodies" are human germline immunoglobulins or reorganized immunoglobulins. For the amino acid sequence encoded by the immunoglobulin gene, The sequence is at least approximately 80%, 81%, 82%, 83%, 84%, 85%, 86%, and 87%. 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% They are 98% or 99% identical. In some cases, "human antibodies" are, for example, Knappi k et al., (2000) J Mol Biol 296:57-86 Consensus framework sequences obtained from human framework sequence analysis, or for example Shi et al., (2010) J Mol Biol 397:385-96 and Human phages presented in international publication WO2009 / 085462 It may contain synthetic HCDR3 incorporated into an immunoglobulin gene library. Furthermore, antibodies derived from non-human species are not included in the definition of "human antibodies."

[0049] "Humanized antibodies" are those in which at least one CDR is derived from a non-human species and at least one F The framework refers to antibodies derived from human immunoglobulin sequences. Humanized antibodies are frameworks. Because the framework can include substitutions, the framework expresses human immunoglobulins. It may not be an exact copy of the germline gene sequence of phosphorus or human immunoglobulin. .

[0050] "Isolated" refers to other components of the system in which molecules are produced, such as in recombinant cells. A homogeneous collection of molecules that have been substantially separated and / or purified from (e.g., synthetic polynucleotides) In addition to rheotide or polypeptide, a tar that has been subjected to at least one purification or isolation step. This refers to protein. "Isolated / isolated" means that it is essentially isolated from other cellular material and / or chemical substances. This refers to molecules not included in the standard, indicating higher purity levels, such as 80%, 81%, 82%, 83%, and 84%. 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% Molecules isolated to a purity of 95%, 96%, 97%, 98%, 99%, or 100% To include.

[0051] "To regulate" means to have a greater response compared to a response mediated by a control or vehicle. Enhanced or reduced test molecules that mediate a response (i.e., downstream effect) It refers to one of the abilities.

[0052] "Natural killer cells" and "NK cells" are used interchangeably and synonymously in this specification. NK cells are CD16 + CD56 + and / or CD57 + TCR - Possesses a phenotype NK cells are differentiated lymphocytes that undergo "self-lysis" through the activation of specific cytolytic enzymes. Ability to bind to and kill cells that cannot express HC / HLA antigens, tumor cells, or NK cell activation The ability to kill other disease cells that express the receptor ligand, and to stimulate or inhibit the immune response. It is characterized by its ability to release protein molecules called cytokines.

[0053] "Specifically bind," "specifically bind," "specifically bound," or "to bind" "A protein molecule is a parent of an antigen or an epitope within that antigen that is related to another antigen." This refers to binding with a higher affinity than typical affinity. Typically, protein molecules bind with approximately 1× 10 -7 M or less, for example, about 5 × 10 -8 M or less, approximately 1×10 -8 M or less, approximately 1×10 -9 M or less, about 1×10 -10 M or less, about 1×10 -11 M or less, or about 1×10 -12 M or less equilibrium dissociation constant (K D ) binds to an antigen or an epitope within the antigen, and typically, the K D is lower than the K D for binding to a non-specific antigen (e.g., BSA, casein) by at le ast 100-fold. In the context of the prostate neoantigens described herein, "specific bind ing" refers to that a proteinaceous molecule binds to the prostate neoantigen without detectab ly binding to the wild-type protein of which the prostate neoantigen is a variant.

[0054] "Tumor cell" or "cancer cell" refers to a cancerous, pre-cancerous, or transformed cell that e xhibits spontaneous or induced phenotypic changes, either in vivo, ex vivo, or in tissue cultu re. These changes do not necessarily involve the uptake of new genetic material. Transform ation can occur through infection with a transforming virus and integration of new genomic n ucleic acid, or uptake of exogenous nucleic acid, it can also occur spontaneously or after e xposure to a carcinogen, which in some cases causes mutation of endogenous genes. Transformati on / cancer is exemplified by morphological changes, cell immortalization, abnormal growth r egulation, focus formation, proliferation, malignant lesions, regulation of tumor-specific marker levels, invasiveness, and tumor growth in a suitable animal host such as nude mice, in vitro, in vivo, and ex vivo.

[0055] As used herein, the term "chimeric antigen receptor" or "CAR" refers to a molecule that compris es an extracellular target-binding domain in a combination that is not naturally found togeth It is defined as a cell surface receptor containing a transmembrane domain and an intracellular signaling domain. This is understood to mean that, in particular, the extracellular domain and intracellular signaling domain are naturally The present invention includes receptors that are not found together in a single receptor protein. Chimeric antigen receptors are mainly found in lymphocytes such as T cells and natural killer (NK) cells. It is intended for use.

[0056] The terms "T cell" and "T lymphocyte" are interchangeable and are used synonymously herein. When used herein, T cells refer to thymocytes, naive T lymphocytes, and immature T cells. Includes lymphocytes, mature T lymphocytes, resting T lymphocytes, or activated T lymphocytes. T cells are, T helper (Th) cells, for example, T helper 1 (Th1) or T helper 2 (Th2) It can be any cell. T cells include helper T cells (HTL; CD4+ T cells), CD4+ T cells, cytotoxic T cells (CTL; ​​CD8+ T cells), tumor-infiltrating cytotoxic T cells (TI L (CD8+ T cells), CD4+CD8+ T cells, or any other subset of T cells It may be. Other exemplary populations of T cells suitable for use in a particular embodiment include: These include naive T cells and memory T cells. Also included are "NKT cells," and these It not only expresses semi-invariant αβ T cell receptors, but also interacts with normal NK cells such as NK1.1. This refers to a special group of T cells that also express various related molecular markers. NKT cells are, NK1.1+ and NK1.1-, as well as CD4+, CD4-, CD8+, and CD8 cells. One example is that the TCR of NKT cells is presented by the MHC I-like molecule CD Id. It is unique in that it recognizes glycolipid antigens. NKT cells are involved in either inflammation or immune tolerance. Due to its ability to produce cytokines that promote certain effects, it may have either a protective or harmful effect. It is possible. It also includes "gamma delta T cells (γδT cells)." Gamma delta T cells This refers to a special group of small subsets of T cells that have different TCRs on their surface. T cells, in which the TCR is composed of two glycoprotein chains, denoted as α and β-TCR chains. This differs from most other factors. The TCR in γδ T cells is composed of a γ chain and a δ chain. T cells can play a role in immune surveillance and immunomodulation, and are important for IL-17 It was found to be a source and to induce an active CD8+ cytotoxic T cell response. Furthermore, it refers to T cells that suppress abnormal or excessive immune responses and play a role in immune tolerance. This also includes "regulatory T cells" or "Tregs." Tregs are typically transcription factors F These are oxp3-positive CD4+ T cells, and also IL-10-producing CD4+ T cells. This may also include juvenile Foxp3-negative regulatory T cells.

[0057] The terms "natural killer cell" and "NK cell" are interchangeable and used herein. Used synonymously. In this specification, NK cells refer to CD16+CD56+ and NK cells refer to differentiated lymphocytes that have the CD57+ TCR phenotype. Activation of vesicular lytic enzymes leads to binding to cells that cannot express "self" MHC / HLA antigens. , the ability to kill, tumor cells or other disease cells that express ligands for NK activation receptors Protein molecules called cytokines have the ability to inflict damage and to stimulate or inhibit the immune response. It is characterized by its ability to release.

[0058] As used herein, the term “antigen” means an antigen to which a T cell receptor can bind. Ingredients (for example, proteins, peptides, polysaccharides, glycoproteins, glycolipids, nucleic acids, etc.) It refers to a molecule (part of or a combination thereof). Antigens can also trigger an immune response. Examples of immune responses include, but are not limited to, antibody production or specific immunological conditions. This may involve activation of pitent cells, or both. Those skilled in the art will know that the antigen never becomes a "gene". Therefore, you will understand that it does not need to be coded. Antigens are produced by synthesis. It may also be derived from a biological sample, or from a macromolecule other than a polypeptide. It is readily apparent that this may also be the case. Such biological samples include tissue. Samples, tumor samples, cells, or other biological components, organisms, proteins / antigens Examples include fluids containing units, killed or inactivated whole cells, or lysates. Yes, but not limited to these.

[0059] The term "antibody" refers to monoclonal antibodies, multispecific antibodies, human antibodies, humanized antibodies, and more. Mela antibody, single-chain Fv(scFv), single-chain antibody, Fab fragment, F(ab') fragment, dys Rufid linkage Fv (sdFv), intrabody, minibody, diabody, and anti- Diotype (anti-Id) antibodies (e.g., anti-Id antibodies against antigen-specific TCRs), and any of the above epitope-binding fragments. The term “antibody” is also used in the U.S. Patent Application Publication. Covalent diamond bodies, such as those disclosed in Patent Publication No. 2007 / 0004909, and Ig- This also refers to DARTS. Antibodies useful as TCR binding molecules include immunoglobulin molecules and Immunologically active fragments of immunoglobulin molecules, i.e., molecules containing antigen-binding sites They can be listed. Immunoglobulin molecules are of any type (e.g., IgG, IgE, IgM, I IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG) 4. IgM1, IgM2, IgA1, and IgA2), or a subclass.

[0060] The term "host cell" refers to any cell containing heterologous nucleic acids. Heterologous nucleic acids are vectors. - (For example, an expression vector) may be used. For example, a host cell produces a substance through the cell. For example, the cellular expression of genes, DNA or RNA sequences, proteins, or enzymes. Any organism selected, modified, transformed, grown, used, or manipulated in any way for the purpose of It can be a cell of origin. The appropriate host can be determined. For example, the host cell is a ve The choice can be based on the cultor skeleton and the desired outcome. For example, plasmids or cosmids It can be introduced into prokaryotic host cells to replicate several types of vectors. Examples of bacterial cells include H5α, JM109, and KCB, but are not limited to these. E(registered trademark) competent cells, and SOLOPACK Gold cells, are used as vectors. It can be used as a host cell for replication and / or expression. Furthermore, E. coli LE3 Bacterial cells such as 92 can be used as host cells for phage viruses. Examples of eukaryotic cells that can be used as cells include yeast (e.g., YPH499, YP Examples include, but are not limited to, H500 and YPH501), insects, and mammals. i. An example of a mammalian eukaryotic host cell for vector replication and / or expression is HeLa. NIH3T3, Jurkat, 293, COS, CHO, Saos, and PC12 were mentioned. These are some examples, but they are not limited to these.

[0061] The host cells of this disclosure contain a DNA or RNA sequence encoding a CAR, Examples include T cells and natural killer cells that express CAR on their cell surface. (Host cell) It enhances T cell activity, natural killer cell activity, cancer treatment, and treatment of autoimmune diseases. It can be used for that purpose.

[0062] "Activation" or "stimulation" means that cells (e.g., T cells and NK cells) are activated as a result. To express sexualization markers, produce cytokines, proliferate, and / or target cells This means inducing a change in the biological state of the cell, which can lead to cytotoxicity. These changes are all This can be caused by the primary stimulus signal. The co-stimulatory signal increases the magnitude of the primary signal. This broadens the range of cells and suppresses cell death after initial stimulation, resulting in a more durable activated state. This leads to a state and, consequently, higher cytotoxicity. "Co-stimulatory signaling" is TCR / CD3 In combination with primary signaling such as eruption, T cell and / or NK cell proliferation And / or upregulation or downregulation of key molecules It refers to the signal that is triggered.

[0063] The term "proliferation" refers to an increase in cell division, whether symmetrical or asymmetrical. The term "expansion" refers to the result of cell division and cell death.

[0064] The term "differentiation" refers to reducing the ability or proliferation of a cell, or limiting the cell's developmental capabilities. This refers to a method of transitioning to a predetermined state.

[0065] The terms "express" and "expression" refer to the generation of information about a gene or DNA sequence. To enable or generate, for example, the transcription and translation of a corresponding gene or DNA sequence. This means generating proteins by activating cellular functions involved in D NA sequences are expressed within or by cells to form "expression products" such as proteins. The expression product itself, for example, the resulting protein, is said to have been "expressed" by the cell. It is also possible to characterize the expression product as intracellular, extracellular, or transmembrane. ru.

[0066] The term "transfection" refers to the process of introducing "foreign" DNA into cells using recombinant DNA technology. In other words, it means introducing exogenous or extracellular nucleic acids. The term "genetic modification" is, "Exotic" (i.e., extrinsic or extracellular) genes, DNA, or RNA to host cells The sequence is introduced, and as a result, the host cell expresses the introduced gene or sequence, thereby producing the desired result. Quality, typically a protein or yeast encoded by an introduced gene or sequence. It means generating a gene or sequence. The introduced gene or sequence is also called "cloned." These are sometimes called "foreign" genes or sequences, and include start, stop, promoter, and sigma. Chimeric antigen receptors such as other sequences used by the genetic mechanisms of the cell, secretion, or other cells. Includes a regulatory or control sequence operably ligated to the encoding polynucleotide. It is permissible. Genes or sequences may contain non-functional sequences that do not have known functions. Host cells that receive and express DNA or RNA are "genetically manipulated." . The DNA or RNA introduced into a host cell comprises cells of the same genus or species as the host cell and may be of any origin, or derived from a different genus or species.

[0067] The term "transduction" refers to the introduction of a foreign nucleic acid into a cell using a viral vector .

[0068] The term "control element" refers to any cis-acting genetic element that controls some aspects of the expression of a nucleic acid sequence . In some embodiments, the term "promoter" essentially comprises the minimal sequence required to initiate transcription . In some embodiments, the term "promoter " comprises a sequence for initiating transcription, and additionally comprises sequences capable of up-regulating or down-regulating transcription, which are generally referred to as "enhancer elements" and "repressor elements", respectively .

[0069] As used herein, "operably linked" and similar phrases, when used with reference to nucleic acids or amino acids, respectively, refer to an operable linkage of nucleic acid sequences or amino acid sequences that are placed in a functional relationship with each other . For example, an operably linked promoter, enhancer element, open reading frame, 5' and 3' UTR, and ter minator sequence result in the correct production of a nucleic acid molecule (e.g., RNA). In some embodiments, operably linked nucleic acid elements result in the transcription of an open reading frame and ultimately the production of a polypeptide (i.e., expression of the open reading frame ). As another example, an operably linked peptide refers to a peptide in which functional domains are arranged at an appropriate distance from each other to confer the intended function of each domain . . .

[0070] "Potentiation", "promotion", "increase", "expansion" or "improvement" generally refers to a vehicle or a control molecule / composition that elicits, induces or causes a greater physiological response (i.e., a downstream effect), as compared to the response caused by either the vehicle or the control molecule / composition. Measurable physiological responses, as will be clear particularly from the understanding of the art and the description herein, can include expansion of T cells, activation, effector function , increased persistence, and / or increased cancer cell killing ability. In certain embodiments, an "increased" or "potentiated" amount can be a "statistically significant" amount, and can be 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 times or more (e.g., 500, 1000 times) the response produced by a vehicle or control composition, and includes such an increase including any integer above 1 and decimal points in between, for example 1.5, 1.6, 1.7, 1.8, etc. composition, and includes such an increase including any integer above 1 and decimal points therebetween, for example 1.5, 1.6, 1.7, 1.8, etc.

[0071] "Decrease", "reduction", "attenuation", "lowering" or "alleviation" generally refers to a vehicle or a control molecule / composition that elicits, induces or causes a smaller physiological response (i.e., a downstream effect), as compared to the response caused by either the vehicle or the control molecule / composition described herein. In certain embodiments, a "decreased" or "reduced ed" amount can be a "statistically significant" amount, and can be 1.1, 1.2, 1.5, 2, 3 the response produced by a vehicle or control composition (reference response), or 1.1, 1.2, 1.5, 2, 3 , 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 times or more (e.g., 500 ​(1000 times) (All integers greater than 1 and decimals in between, e.g., 1.5, 1.6, This may include decreases such as 1.7, 1.8, etc.

[0072] The term "effective" applied to dosage or amount refers to the amount administered to the target who needs it. This refers to the amount of compound or pharmaceutical composition sufficient to produce the desired activity. (Combination of active ingredients) When administering these, the effective dose of the combination is the effective dose of each component that would be effective if administered individually. Please note that the amount may or may not be included. The required dosage depends on the species, age, and overall health of the subject, the severity of the condition being treated, and the specific medication used. The appropriate course of action varies depending on the drug, the method of administration, and the individual patient.

[0073] When used in relation to the compositions described herein, the phrase "pharmaceutically acceptable" means Physiologically tolerable and typically does not cause adverse reactions when administered to mammals (e.g., humans). This refers to the molecular entities and other components of such compositions that do not cause the formation of such entities. Preferably, the term "medical "Pharmacologically acceptable" means that for use in mammals, more specifically in humans, it has been approved by the federal government. Alternatively, it may be authorized by a state government regulatory agency, or it may be regulated by the United States Pharmacopeia or other general regulations. It means that it is listed in the officially recognized pharmacopoeia.

[0074] The term "protein," as used herein, includes protein fragments of all lengths. In addition to natural and synthetic proteins, fusion proteins, and glycoproteins, all other Types of modified proteins (e.g., phosphorylation, acetylation, myristoylation, palmitoylation) Formulation, glycosylation, oxidation, formylation, amidation, polyglutamylation, ADP-ribosyl modifications including, but not limited to, proteins obtained from phosphorylation, PEGylation, biotinylation, etc. encompasses all types of proteins.

[0075] Unless otherwise specifically stated, the terms "nucleic acid", "nucleotide", and "polynucleotide" encompass both DNA and RNA. The term "nucleic acid sequence" or "nucleotide sequence" refers to a nucleic acid sequence that encodes an amino acid, and these terms may also refer to a nucleic acid sequence comprising a portion encoding any amino acid added as a cloning artifact, including any amino acid encoded by a linker.

[0076] The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil and sesame oil. Water, aqueous solutions, saline, as well as dextrose and glycerol aqueous solutions are particularly preferably used as carriers for injectable solutions. Alternatively, the carrier can be a solid dosage form carrier including, but not limited to, one or more of binders (for compressed pills), glidants, encapsulating agents, flavoring agents, and coloring agents. Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin.

[0077] The term "about" or "approximately" includes being within a statistically meaningful range of the value. Such a range can be within one order of magnitude of a given value or range, preferably within 50%, more preferably within 20%, still more preferably within 10%, and even more preferably within The acceptable variation encompassed by the words "approximately" or "about" is a specific system under study. It depends on the element and can be easily understood by those skilled in the art.

[0078] "Kallikrein-related peptidase 2", "hK2", or "klk2" refers to kallikrein This refers to a known protein also called In-2, granular kallikrein 2, or HK2. K2 is produced as a preproprotein and is cleaved during proteolysis to activate the proteoprotein. It generates an enzyme. All hK2 isoforms and variants are included in "hK2". The amino acid sequences of various isoforms are listed by GenBank accession number NP. _005542.1, NP_001002231.1, and NP_001243009 It is searchable. The full-length amino acid sequence of human hK2 in SEQ ID NO: 62 is shown. It contains a signal peptide (residues 1-18) and a propeptide region (residues 19-24). .

[0079] The terms used herein are for the sole purpose of describing specific embodiments. This is not intended to be an exhaustive limitation. When used herein, the context may be otherwise clearly indicated. Unless otherwise specified, the indefinite articles "a," "an," and "the" are considered to refer to multiple things. It should be explained.

[0080] Chimeric antigen receptor The present invention generally involves genetically modifying a gene to stably express a desired chimeric antigen receptor. Regarding the use of T cells. Chimeric antigen receptors (CARs) are T cell signaling domains. An artificially constructed hive containing the antigen-binding domain of an antibody (scFv) linked to it. It is a lid protein or polypeptide. A key characteristic of CARs is that they are selected non-MHC restricted. Redirecting T cell specificity and reactivity toward the selected target, monoclonal antibody One notable feature is its ability to utilize its antigen-binding properties. Through recognition of non-MHC restriction antigens, C T cells that express AR can recognize antigens independently of antigen processing. Therefore, it bypasses the main mechanism of tumor escape. Furthermore, when expressed in T cells, C AR is favored by endogenous T cell receptor (TCR) alpha and beta chains. It does not dimerize.

[0081] The CAR described herein comprises at least an extracellular antigen-binding domain and a transmembrane domain. intracellular signals containing functional signaling domains derived from the following stimulating molecules The signaling domain (also referred to herein as the "cytoplasmic signaling domain") includes This invention provides a recombinant polypeptide construct. T cells expressing CAR are described in this specification. In the book, these are referred to as CAR T cells, CAR-T cells, or CAR-modified T cells, and their use The terms are used interchangeably in this specification. Cells have antibody-binding domains on their surface. It can be genetically modified to ensure stable expression and exhibits novel MHC-independent antigen specificity. Grant it.

[0082] In some cases, T cells recognize the antigen recognition domain of a particular antibody as being CD3-zeta or F C combines with the intracellular domain of the cγRI protein to form a single chimeric protein. The gene is genetically modified to stably express AR. In one embodiment, the stimulating molecule is T This is a zeta chain associated with cell receptor complexes.

[0083] "Intracellular signaling domain" or "cytoplasmic signaling domain" is defined herein as follows: When used, it refers to the intracellular portion of the molecule. This is because it generates secondary signaling molecules. To control cellular activity through specific signaling pathways, information is transmitted within the cell. The effector acts by, or responds to such information-transmitting substances. It is the functional part of a protein that acts by functioning in a specific way. The domain enhances the immune effector function of CAR-containing cells (e.g., CAR-T cells). It generates signals. For example, consider the immune effector function in CAR-T cells. These include cell-lytic activity and helper activity (including cytokine secretion).

[0084] In one embodiment, the intracellular signaling domain is the primary intracellular signaling domain. It may include. Examples of primary intracellular signaling domains include primary stimulus or antigen-dependent blemishes. Examples include those derived from molecules involved in sigma. In one embodiment, intracellular sigma The signaling domain may include a co-stimulatory intracellular domain. Examples of "ins" include those derived from molecules involved in co-stimulatory signals or antigen-independent stimuli. For example, in the case of CAR-T cells, the primary intracellular signaling domain is located in the T cell. The receptor may contain a cytoplasmic sequence, and the co-stimulatory intracellular signaling domain may be a co-receptor or co-stimulatory domain. It may contain cytoplasmic sequences derived from highly active molecules.

[0085] The primary intracellular signaling domain is an immune receptor tyrosine-based activation motif or It may contain a signaling motif known as ITAM. Primary cytoplasmic signaling sequence Examples of ITAMs containing CD3-zeta, FcR-gamma, FcR-beta, and CD3-zeta are available. CD3 Gamma, CD3 Delta, CD3 Epsilon, CD5, CD22, CD79a, CD79 b. and CD66d DAP10 and DAP12 are examples of those derived from these, It is not limited to them.

[0086] "Zeta" or alternatively, "Zeta chain," "CD3-zeta," or "TCR-zeta" The term is provided as GenBank accession number BAG36664.1. Proteins, or non-human species, such as mice, rabbits, primates, rodents, and monkeys. Defined as equivalent residues from apes, etc., the "zeta-stimulating domain" or alternatively, The "CD3-zeta-stimulating domain" or "TCR-zeta-stimulating domain" is involved in T cell activation. Sufficient zeta chain cytoplasmic domains are needed to functionally transmit the necessary initial signals. It is defined as a amino acid residue. Sometimes, for example, in the embodiments and / or aspects of this disclosure The "zeta-stimulating domain" or "CD3-zeta-stimulating domain" is the "primary signaling domain." It is also called the "transformation domain". In one embodiment, the cytoplasmic domain of the CD3 zeta-stimulating domain is , residues 52-164 of GenBank accession number BAG36664.1, or Functional orthologues of non-human species, such as mice, rodents, monkeys, and apes. It contains equivalent residues derived from it. In a preferred embodiment, the intracellular signaling domain is C Contains a D3 zeta-stimulating domain. In one embodiment, "zeta-stimulating domain" or "CD3-zeta-stimulating domain". The "stimulatory domain" is the sequence provided as SEQ ID NO: 28, or less than SEQ ID NO: 28. 50, at least 55, at least 60, at least 65, at least 70, less 75, at least 80, at least 85, at least 90, at least 91, less 92, at least 93, at least 94, at least 95, at least 96, less Both sequences have 97%, at least 98%, or at least 99% sequence identity. .

[0087] The term "costimulatory molecule" refers to a molecule that specifically binds to a costimulatory ligand, thereby affecting T cells. Co-stimulatory responses (e.g., proliferation, but not limited to) mediated by allogeneic tumors on T cells This refers to a co-stimulatory partner. A co-stimulatory molecule is an antigen receptor or its ligament necessary for an efficient immune response. These are cell surface molecules other than brands. Co-stimulatory molecules include MHC class I molecules, BTLA, and Toll ligand receptors, as well as OX40, CD2, CD27, CD28, CDS, ICAM-1, LFA-1 (CD11a / CD18), and 4-1BB (CD137) These are some examples, but are not limited to them.

[0088] The co-stimulatory intracellular signaling domain can be the intracellular portion of a co-stimulatory molecule. The children belong to the following protein families: TNF receptor proteins, immunoglobulin-like proteins. Calcium, cytokine receptor, integrin, signal transduction lymphocyte activating molecule (SLAM) It can be represented as a protein, and as an activated NK cell receptor. Examples of such molecules include For example, CD27, CD28, 4-1BB (CD137), OX40, GITR, CD30 MyD88, CD40, ICOS, BAFFR, HVEM, Lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, SLAMF7, NKp80 Examples include ligands that specifically bind to CD160, B7-H3, and CD83. .

[0089] An intracellular signaling domain is the entire intracellular portion of the molecule from which it originates, or a native molecule. This may include entire intracellular signaling domains or functional fragments thereof.

[0090] The term "4-1BB" or alternatively "CD137" refers to GenBank accession The amino acid sequence provided as number AAA62478.2, or for non-human species, e.g., Mau The TNFR superfamily has equivalent residues from rodents, monkeys, apes, and other animals. The "4-1BB Co-Stimulation Domain" refers to the members of the GenBank accession number. Amino acid residues 214-255 of AAA62478.2, or non-human species, e.g., mouse It is defined as equivalent residues from rodents, monkeys, and apes. Sometimes, for example, In aspects and / or embodiments of this disclosure, the "4-1BB co-stimulation domain" is defined as the "co-stimulation domain". It is also called "main."

[0091] In a preferred embodiment, the intracellular signaling domain is the 4-1BB costimulatory domain or It contains a co-stimulatory intracellular signaling domain called the "CD137 co-stimulatory domain". In this case, the "4-1BB co-stimulatory domain" or "CD137 co-stimulatory domain" is the sequence number. The sequence provided as 27, or non-human species, e.g., mouse, rodent, monkey, ape Equivalent residues from any of the above, or at least 50, at least 55, at least from SEQ ID NO: 27 60, at least 65, at least 70, at least 75, 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 less The sequence has at least 99% sequence identity. In a preferred embodiment, 4-1BB together The stimulating domain is the sequence provided as sequence number 27.

[0092] In one embodiment, a transmembrane domain naturally associated with one of the domains within the CAR is used. In another embodiment, the transmembrane domain is selected or modified by amino acid substitution, and To avoid binding such domains to the transmembrane domains of the same or different surface membrane proteins, Interaction with other members of the receptor complex can be minimized. One example of implementation. In this state, the transmembrane domain includes the CD8α hinge domain. In one embodiment, CAR is It includes the CD8α hinge domain and the CD8α transmembrane domain.

[0093] In some embodiments, the cytoplasmic signaling domain is less than the definitions provided herein. It further includes one or more functional signaling domains derived from at least one co-stimulatory molecule. In one embodiment, the co-stimulatory molecules are 4-1BB (i.e., CD137), CD27, CD Selected from 3-zeta and / or CD28. CD28 plays a significant role in T cell costimulation. It is an important T cell marker. CD27 is a member of the tumor necrosis factor receptor superfamily. It is a bar and functions as a co-stimulatory immune checkpoint molecule. 4-1BB acts on T cells. It transmits powerful co-stimulatory signals, promotes differentiation, and enhances the long-term survival of T lymphocytes. 3-zeta associates with the TCR to generate a signal and activate the tyrosine-based immune receptor. Contains motifs (immunoreceptor tyrosine-based activation motifs, ITAMs) In another embodiment, the co-stimulatory molecule is MyD88 or CD40.

[0094] In one embodiment, the CAR comprises an intracellular hinge domain containing CD8 and CD28, 4-1 It includes BB and an intracellular T cell receptor signaling domain containing CD3-zeta. In another embodiment, CAR comprises an intracellular hinge domain and CD28, 4-1BB, and C The D3 zeta-containing intracellular T cell receptor signaling domain is included, and the hinged domain The antibody contains all or part of the extracellular region of CD8, CD4, or CD28, or the entire constant region of the antibody. or part of, FcRIIIA receptor, IgG hinge, IgM hinge, IgA hinge, Ig Includes all or part of a D hinge, IgE hinge, or Ig hinge. The IgG hinge is Ig G1, IgG2, IgG3, IgG4, IgM1, IgM2, IgA1, IgA2, Ig It may be derived from D, IgE, or a chimera thereof.

[0095] The CAR described herein comprises at least an extracellular antigen-binding domain and a transmembrane domain. For example, intracellular signals containing functional signaling domains derived from the following stimulating molecules The signaling domain (also referred to herein as the "cytoplasmic signaling domain") and The invention provides a recombinant polypeptide construct that includes ,

[0096] In one embodiment, the CAR comprises an extracellular antigen recognition domain, a transmembrane domain, and a stimulating molecule. An intracellular signaling domain containing a functional signaling domain derived from and containing It contains a MELLA fusion protein. In one embodiment, the CAR has an extracellular antigen recognition domain and a membrane The transconducting domain, the functional signaling domain derived from the co-stimulator molecule, and the domain derived from the stimulator molecule. An intracellular signaling domain containing a functional signaling domain, and a chimeric fusion domain containing It contains a combined protein. In one embodiment, the CAR has an extracellular antigen recognition domain and a transmembrane domain. Main and at least two functional signaling domains derived from one or more co-stimulatory molecules Intracellular signaling domains containing functional signaling domains derived from ions and stimulating molecules It contains a chimeric fusion protein that includes n and .

[0097] The CAR of the present invention contains only the CD28 and / or 4-1BB signaling domain. To such effect, or any other desired cytoplasmic domain useful in the context of the CAR of the present invention They may be designed to be combined. In one embodiment, the cytoplasmic domain of CAR is CD It may further include the signaling domain of 3-zeta. For example, the cytoplasmic domain of CAR and This includes the CD3-zeta, 4-1BB, and CD28 signaling modules, as well as These are some possible combinations, but are not limited to them.

[0098] In one embodiment, CAR has a hinge domain including a CD8α hinge domain, and CD8 α-transmembrane domain and intracellular signaling domains including 4-1BB and CD3-zeta and, including. In one embodiment, CAR is a hinge domain including the CD8α hinge domain. The cells containing the CD8α transmembrane domain, CD28, 4-1BB, and CD3-zeta It includes an internal signaling domain.

[0099] In another embodiment, the CAR comprises a CD8α hinge domain and a CD8α transmembrane domain. , intracellular zeta, including the CD3-zeta signaling domain and the 4-1BB costimulatory domain Includes the Gnar transmission domain.

[0100] In another embodiment, the CAR is a CD8a-hinge region containing the amino acid sequence of SEQ ID NO: 25. The region, the transmembrane domain containing the amino acid sequence of SEQ ID NO: 26, and the amino acid sequence of SEQ ID NO: 27 Co-stimulatory domains including columns and primary signaling domains including the amino acid sequence of SEQ ID NO: 28 It includes an intracellular signaling domain containing n, and also contains.

[0101] Therefore, the present invention proposes CAR T cells and methods for their use in adoptive therapy. To provide.

[0102] This disclosure relates to the CARs, nucleic acids, polypeptides, and proteins described herein. It further provides variants (e.g., functional variants). A "variant" is, for example, a replacement or insertion. or one or more modifications, such as deletion, to the reference polypeptide or reference polynucleotide Refers to a polypeptide or polynucleotide different from the above. When used herein, "functional The term "variant" refers to a variant that is substantially or significantly different from the parent CAR, polypeptide, or protein. This refers to CARs, polypeptides, or proteins that have sequence identity or similarity, and these A functional variant is a variant of a CAR, polypeptide, or protein. It retains activity. Functional variants include, for example, the CARs and polypeptides described herein. , or encompassing variants of proteins (parent CAR, polypeptide, or protein), Labeled to the same degree, the same degree, or a higher degree as the parent CAR, polypeptide, or protein. It retains the ability to recognize target cells. With respect to parent CARs, polypeptides, or proteins, Functional variants are, for example, amides of parent CAR, polypeptide, or protein. The no-acid sequence accounts for at least approximately 30%, 40%, 50%, 60%, 75%, and 80% of the total. Approximately 85%, approximately 90%, approximately 91%, approximately 92%, approximately 93%, approximately 94%, approximately 95%, approximately 96%, They may be approximately 97%, 98%, 99%, or more identical.

[0103] f) In this specification, the structure of a polypeptide is defined as having an enumerated reference sequence (with a given sequence number). It is appropriately defined based on the sequence identity % with ( ). In this regard, two amino acids The % sequence identity between sequences is calculated by comparing these two sequences that have been aligned in the most optimal way. This can be determined by comparing these two amino acid sequences. This may involve additions or deletions to the reference sequence for optimal alignment between the sequences. The percentage of uniformity is determined by finding the number of identical amino acid residues at the same position between two sequences. Divide the number of identical positions by the total number of positions in the comparison window, and multiply the result by 100. This is calculated by obtaining the percentage of identity between these two sequences. Typically, the comparison window corresponds to the total length of the sequences being compared. For example, BLAS T Program, "BLAST 2 Sequence" (Tatusova et al, "Blast "A novel tool for comparing 2-sequence protein and nucleotide sequences," FEMS It is possible to use Microbiol Lett. 174:247-250. The website is http: / / www.ncbi.nlm.nih.gov / gorf / bl 2.html is available, and the parameters used are the default (specifically, Lameta "Open Gap Penalty": 5, and "Extension Gap Penalty" "Ti": 2; The selected matrix is, for example, the matrix proposed by the program These are parameters specified in the "BLOSUM 62" tool, and the two parameters being compared are... The percentage of identity between sequences is calculated directly by the program. Determining sequence identity for a given sequence is within the scope of the skills of a person skilled in the art, and BLAST (trademark) This can be done using commercially available analysis software such as [mention specific software name].

[0104] A functional variant is, for example, a parent CAR having at least one conserved amino acid substitution. , may include a polypeptide or protein amino acid sequence. In another embodiment, functional The variant is a parent CAR, polypeptide having at least one non-conservative amino acid substitution. , or may include the amino acid sequence of a protein. In this case, non-conservative amino acid substitutions are functional. It is not possible to suppress or inhibit the biological activity of the target variant. This allows for the improvement of the biological activity of the functional variant, and as a result, the functional variant The biological activity of the parent CAR, polypeptide, or protein is increased compared to the parent CAR.

[0105] The amino acid substitutions of CAR in the present invention may be conservative amino acid substitutions. The exchange is known in the art and has specific physical and / or chemical properties. Replace one amino acid with another amino acid that has the same or similar chemical or physical properties. This includes amino acid substitutions. For example, a conservative amino acid substitution is an acid that is substituted with another acidic amino acid. A sexual amino acid (e.g., Asp or Glu), substituted with another amino acid having a nonpolar side chain. Amino acids having nonpolar side chains (e.g., Ala, Gly, Val, Ile, Leu, M) (e.g., et, Phe, Pro, Trp, Val), basic amino acids substituted with other basic amino acids Amino acids (Lys, Arg, etc.), polar side chains substituted with other amino acids that have polar side chains These may be amino acids that contain (Asn, Cys, Gln, Ser, Thr, Tyr, etc.) ru.

[0106] CARs, polypeptides, or proteins are defined by a specific amino acid sequence or as described herein. Essentially, the sequence can become, and as a result, other components, such as other amino acids, can become functional. It does not substantially alter the biological activity of riantoxin.

[0107] CARs, polypeptides, and proteins (functional portions and functional parts) of embodiments of the present disclosure (including riant) can be of any length, that is, it can contain any number of amino acids, However, CAR, polypeptide, or protein (or functional part or its functional variant) Ants possess biological activities such as the ability to specifically bind to antigens and disease cells within the host. The ability to detect cells (e.g., cancer cells), or the ability to treat or prevent diseases within a host. It maintains, for example, a polypeptide has a length of approximately 50 to 5000 amino acids, for example, Approximately 50, approximately 70, approximately 75, approximately 100, approximately 125, approximately 150, approximately 175, approximately 200, approximately 22 5, approximately 250, approximately 275, approximately 300, approximately 325, approximately 350, approximately 375, approximately 400, approximately 42 5, approximately 450, approximately 475, approximately 500, approximately 525, approximately 550, approximately 575, approximately 600, approximately 62 5, approximately 650, approximately 675, approximately 700, approximately 725, approximately 750, approximately 775, approximately 800, approximately 82 5. Approximately 850, 875, 900, 925, 950, 975, 1000 or so The amino acid length may be greater than or equal to this. The polypeptide of the present invention also includes oligopeptides.

[0108] CAR, polypeptide, and protein according to embodiments of the present invention (functional portion and (Including functional variants) are synthetic amino acids instead of one or more naturally occurring amino acids. It may contain an acid. Such synthetic amino acids are known in the art, for example, For example, aminocyclohexanecarboxylic acid, norleucine, α-aminon-decanoic acid, ho Moserine, S-acetylaminomethyl-cysteine, trans-3- and trans-4- Hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, α- (2-amino-2-norbornane)-carboxylic acid, α,γ-diaminobutyric acid, α,β-diaminobutyric acid Minopropionic acid, homophenylalanine, 4-chlorophenylalanine, 4-carbboxy Phenylalanine, β-phenylserine, β-hydroxyphenylalanine, phenyl Lysine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, N '-benzyl-N'-methyllysine, N',N'-dibenzyllysine, 6-hydrox Cyridine, ornithine, α-aminocyclopentanecarboxylic acid, α-aminocyclohexa α-aminocycloheptanecarboxylic acid, indoline-2-carboxylic acid, 1 ,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, amino Examples include monoamide ronate and α-tert-butylglycine.

[0109] CAR, polypeptide, and protein (functional portion and functional portion) according to embodiments of the present invention (including riantos) can undergo post-translational modifications. These include glycosylation, esters. tellation, N-acylation, amidation, carboxylation, phosphorylation, esterification, for example, di It can be cyclized via sulfide bridges, or converted into an acid addition salt. In some embodiments, these are dimerized, polymerized, or conjugated. It is being done.

[0110] CARs, polypeptides, and / or proteins (functional portions and so) according to embodiments of the present invention (including functional variants) can be obtained by methods known in the art. Suitable methods for the de novo synthesis of polypeptides and proteins can be found in the references, e.g., C han et al.,Fmoc Solid Phase Peptide Synt hesis,Oxford University Press,Oxford,Uni ted Kingdom,2000;Peptide and Protein Dru g Analysis, ed. Reid, R., Marcel Dekker, Inc. ,2000;and Epitope Mapping,ed.Westwood et. al.,Oxford University Press,Oxford,Unit It is described in ed. Kingdom, 2001. Also, polypeptides and proteins. This involves recombinantly using the nucleic acids described herein with a standard recombination method. It can be produced. For example, Sambrook et al., Molecular Clo ning:A Laboratory Manual,3rd ed.,Cold Sp ring Harbor Press, Cold Spring Harbor, NY .2001; and Ausubel et al.,Current Protocols in Molecular Biology,Greene Publishing Associates and John Wiley & Sons, NY, 1994 See . Furthermore, the CAR, polypeptide, and protein (functional portion) of the present invention Some of these (including their functional variants) are sources of plants, bacteria, insects, mammals, etc. It can be isolated and / or purified. Methods for isolation and purification are available in the art. This is known. Alternatively, the CARs, polypeptides, and / or proteins described herein The qualities (including functional parts and their functional variants) can be commercially synthesized. In this regard, CARs, polypeptides, and proteins are synthesized, recombinant, isolated, and It can be purified or cleansed.

[0111] To produce recombinant nucleic acids used to generate the polypeptides described herein Examples of modified nucleotides that can be used for this purpose include 5-fluorouracil, 5- Bromouracil, 5-Chloruracil, 5-Iodouracil, Hypoxanthine, Xanthin n, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, carboxy Methylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, Dihydrouracil, N 6 - Substituted adenine, 7-methylguanine, 5-methylaminomethyl Uracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylcu -osin, 5"-methoxycarboxymethyluracil, 5-methoxyuracil, 2-meth Lucio-N 6 -Isopentenyl adenine, uracil-5-oxyacetic acid (v), wibutox wybutoxosine, pseudouracil, cuosine, beta-D-galactosyl Quosin, Inosin, N6 -Isopentenyl adenine, 1-methylguanine, 1-methylguanine Tylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, 2-thiocytosine, 5-methyl-2-thio Lacil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-ol Methyl xyacetate, 3-(3-amino-3-N-2-carboxypropyl)uracy Examples include, but are not limited to, ru and 2,6-diaminopurine.

[0112] Nucleic acids are CARs, polypeptides, proteins, or functional parts or their functions. Includes any isolated or purified nucleotide sequence encoding any of the variants It is possible. Alternatively, the nucleotide sequence is a degenerate nucleotide sequence of any of the sequences. Alternatively, it may include combinations of degenerate arrangements.

[0113] Some embodiments of the present invention also provide isolated or purified nucleic acids, wherein the nucleic acids are Nucleotide sequences that are complementary to any of the nucleotide sequences of the nucleic acids described in the specification. or stringent elements in any nucleotide sequence of the nucleic acids described herein The item contains a nucleotide sequence that hybridizes below.

[0114] Nucleotide sequences that hybridize under stringent conditions are highly stringent. Hybridization is possible under stringency conditions. "High stringency conditions" refers to nucleotides. The sequence is detected in a stronger amount than nonspecific hybridization in the target sequence (as specified herein). To specifically hybridize to any of the nucleotide sequences of the nucleic acids listed in [the relevant section]. This means that high stringency conditions include polynucleotides with precisely complementary sequences. Otid, or a polynucleotide containing a small amount of scattered mismatches, nucleotid The resulting sequence had several small regions (e.g., 3-12 bases) that matched the original sequence. The conditions for identification from the DOM sequence are as follows: Such complementary subregions are 14-17 or so It melts more easily than the full-length complement of bases greater than this, and is a highly stringent hybrid. Illumination makes them easily identifiable. Relatively high stringency For example, low-salt and / or high-temperature conditions (approximately 0.02-0 at temperatures of approximately 50-70°C) Examples include providing 0.1M NaCl or equivalent. The agency condition is a mismatch between the nucleotide sequence and the template or target strand. Even if present, it is hardly tolerable, and the expression of any of the CARs described herein is not detectable. It is particularly suitable for this purpose. The conditions are such that by gradually increasing the amount of formamide added, It is generally understood that they can become agents.

[0115] In one embodiment, the nucleic acid of the present invention can be incorporated into a recombinant expression vector. The disclosure provides recombinant expression vectors comprising any of the nucleic acids of the present invention. When used, the term "recombinant expression vector" refers to mRNA expressed by host cells, and tannins. Genetically modified oligonucleotides that enable the expression of proteins, polypeptides, or peptides. This refers to a creotide or polynucleotide construct, in which case the construct is , nucleotide sequences encoding mRNA, proteins, polypeptides, or peptides The vector contains and expresses mRNA, protein, polypeptide, or peptide within cells. The vectors described herein are brought into contact with the cells under conditions sufficient to allow them to do so. It does not exist naturally. However, some vectors may exist naturally. The recombinant expression vectors used are DNA and RNA (which may be single-stranded or double-stranded and synthesized). Nucleotides may be derived from, or partially from, natural sources, and may be natural, non-natural, or modified. It may contain any type of nucleotide, including but not limited to (which may contain nucleotides). Recombinant expression vectors are naturally occurring nucleotide linkages, or naturally occurring This may include internucleotide ligations that do not occur, or ligations of both types. It is not naturally occurring or Modified nucleotides or internucleotide links do not inhibit the transcription or replication of the vector. stomach.

[0116] In one embodiment, the recombinant expression vector of the present invention is any suitable recombinant expression vector - It may be used to transform or transfect any suitable host. As a suitable vector, for the reproduction and proliferation of plasmids and viruses, Examples include vectors designed for expression or both. The vectors are p UC Series (Fermentas Life Sciences, Glen Burn) i.e., Md.), pBluescript series (Stratagene, LaJol la, Calif.), pET series (Novagen, Madison, Wis.) pGEX series (Pharmacia Biotech, Uppsala, Swed (en), and the pEX series (Clontech, Palo Alto, Calif.) The group consisting of λGT10, λGT11, λEMBL4, and λNM11 can be selected. 49. Using bacteriophage vectors such as λZapII (Stratagene) This is possible. Examples of plant expression vectors include pBI01, pBI01.2, and pB Examples include I121, pBI101.3, and pBIN19 (Clontech). Examples of expression vectors include pEUK-Cl, pMAM, and pMAMneo(Cl). Examples include ntech. Recombinant expression vectors are viral vectors, for example, letho It could be a virus vector, such as a gamma retrovirus vector.

[0117] In one embodiment, the recombinant expression vector of the present invention is, for example, the Sambrook vector described above. The standard set described in et al. (above) and Ausubel et al. (above) Prepared using replacement DNA technology. Constructs circular or linear expression vectors. The kut can be prepared to contain a replication system that functions in a prokaryotic or eukaryotic host cell. The replication system is, for example, ColE1, SV40, 2μ plasmid, λ, bovine papilloma. It can be derived from words like Rus.

[0118] Recombinant expression vectors are either DNA-based or RNA-based, as appropriate. Considering the type of host into which the vector will be introduced (e.g., bacteria, plants, fungi, or animals), It may include regulatory sequences specific to the substance, such as start and stop codons for transcription and translation.

[0119] Recombinant expression vectors allow for the selection of transformed or transfected hosts. It may contain one or more marker genes that cause biocide resistance (for example). (resistance to antibiotics, heavy metals, etc.), nutritional requirements, to provide protonutrient in the host. This includes complementation of the expression vector, etc. Suitable marker genes for the expression vector described include, for example, Neomycin / G418 resistance gene, histidinol x resistance gene, histidinol resistance Examples include genes, tetracycline resistance genes, and ampicillin resistance genes.

[0120] Recombinant expression vectors are CARs, polypeptides, or proteins (functional portions and The nucleotide sequence encoding (including its functional variant), or CAR, polyp A nucleotide sequence that is complementary to or related to a nucleotide sequence encoding a protein. A nucleotide sequence that hybridizes to this is operably linked to a natural or normative nucleotide. May include a promoter. Promoter selection (e.g., strong, weak, tissue-specific, inducible) , and development-specific effects are within the scope of the art of those skilled in the art. Similarly, promoting nucleotide sequences Combining it with a promoter is also within the scope of the skills of those skilled in the art. The promoter is nonviral. Promoter or virus promoter, for example, cytomegalovirus (cytomegalovirus) RSV promoter, RSV promoter, SV40 promoter, or mouse promoter It could be a promoter found in long terminal repeats of stem cell viruses.

[0121] Recombinant expression vectors are used for either transient expression or stable expression, or both. It can be designed for both constitutive and inducible expression. Recombinant expression vectors can also be used for constitutive or inducible expression. It can be manufactured for current purposes.

[0122] Furthermore, recombinant expression vectors can be constructed to contain suicide genes. When used, the term "suicide gene" refers to a gene that causes cells to die by expressing a suicide gene. It refers to a gene. A suicide gene is a gene that causes cells expressing that gene to react to a drug (e.g., a drug). It confers sensitivity, causing cells to die when they come into contact with or are exposed to the drug. It may be a gene that causes suicide. Suicide genes are known in the field of technology, for example, Herpes simplex virus (HSV) thymidine kinase (TK) gene, cytosine deaminator Examples include ze, purine nucleoside phosphorylase, and nitroreductase.

[0123] CAR, polypeptide, or protein (either its functional part or variant) (including), host cells, nucleic acids, recombinant expression vectors, host cell populations, or antibodies, or A conjugate containing any of its antigen-binding sites, for example, a bioconjugate is , included within the scope of the present invention. Conjugates and methods for synthesizing conjugates are generally Well, it is known in the relevant technical field (for example, Hudecz, F., Methods) Mol.Biol.298:209-223(2005)and Kirin et a See l., Inorg Chem. 44(15):5405-5415 (2005). I want to be.

[0124] One embodiment of the present invention involves binding to the epitope of the CAR of the present invention, for example, specifically binding The present invention further provides an antibody or its antigen-binding portion.

[0125] The antibody may be any type of immunoglobulin known in the art. Globulins are classified into five main classes: IgA, IgD, IgE, IgG, and Ig M can be assigned to IgA and IgG, which are isotypes IgA1, IgA2, and IgG. 1. Further subdivided into IgG2, IgG3, and IgG4. The antibody light chains of vertebrate species are: Based on the amino acid sequences of those constant domains, two types are identified: kappa (κ) and lambda (λ). It can be assigned to one of the types. The antibody is of any class or isotype. obtain.

[0126] Antibodies include monoclonal antibodies, including mouse, human, humanized, and chimeric monoclonal antibodies. Body, polyclonal, antigen-binding fragment, bispecific or multispecific antibody, monomer, dimer, Tetramer or multimer antibodies, single-chain antibodies, domain antibodies, and antigenic antibodies of the required specificity. Immunoglobulin molecules containing any other modified stereochemistry including the fusion site, It contains molecules. Antibodies are naturally occurring antibodies, for example, mammalian antibodies (e.g., mice, primates). Isolation and / or from animals (such as mice, rabbits, goats, horses, chickens, hamsters, and humans) This could be a purified antibody. Alternatively, the antibody may be genetically engineered (e.g., genetically engineered). It could be a manipulated antibody.

[0127] Humanized antibodies have antigen-binding sites derived from non-human species, and the variable region framework is, Derived from human immunoglobulin sequences. Human antibodies possess both a framework and an antigen-binding site. The latter has heavy chain variable regions and light chain variable regions derived from human-derived sequences.

[0128] Furthermore, antibodies possess any level of affinity or binding activity to the functional portion of CAR. Obtain. In some embodiments, the antibody has an affinity (K) within a certain range. D ) binds to the hK2 antigen. This is possible. In one embodiment of the present invention, and in each of the numbered embodiments listed below, The embodiments and some of the embodiments are practiced by those skilled in the art. When determined by surface plasmon resonance or Kinexa method, antibodies exhibit high affinity, for example, , about 10 -7 For KD below M, for example, not limited to, but 1-9.9 (or 1, (2, 3, 4, 5, 6, 7, 8, or 9, or any of the values ​​among them) × 10 - 8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 M, 10 - 14 M, 10 -15 M, or any range or value within it, K D It then binds to the hK2 antigen. An example of affinity is 1 × 10 -8 It is less than or equal to M. Another example of affinity is 1 × 10 -9 More than M It is below.

[0129] A method for testing an antibody's ability to bind to any functional part of a CAR is described in the technical section. Known in the field, and applicable to any antibody-antigen binding assay, e.g., radioimmunoassay. (radioimmunoassay, RIA), Western blot, enzyme-coupled immunosorbent assay (enzyme) ME-linked immunosorbent assay (ELISA), immunoprecipitation, and competitive inhibition assays, etc. These are some examples.

[0130] Suitable methods for producing antibodies are known in the art. For example, standard methods The ibridomas method is, for example, described by Koehler and Milstein, Eur.J. Immunol.,5,511-519(1976),Harlow and Lane (eds.), Antibodies: A Laboratory Manual, CS H Press (1988), and CA Janeway et al. (eds.) ,Immunobiology,5th Ed.,Garland Publishing g, New York, NY (2001)) is described. Alternatively, other methods For example, the EBV-hybridoma method (Haskard and Archer, JI mmunol.Methods,74(2),361-67(1984),and Ro der et al.,Methods Enzymol.,121,140-67(1 986)), and bacteriophage vector expression systems (e.g., Huse et al.) (See Science, 246, 1275-81 (1989)) but the relevant technical portion It is known in the wild. Furthermore, methods for producing antibodies in non-human animals are known, for example, in the United States. Patent Nos. 5,545,806, 5,569,825, and 5,714,352 This is described in the patent number and in U.S. Patent Application Publication No. 2002 / 0197266(A1).

[0131] It is also possible to generate antibodies using phage display. In this regard, antibodies Phage libraries encoding antigen-binding variable (V) domains are standard molecular biological It can be produced using scientific and recombinant DNA technology (e.g., Sambrook See et al. (above) and Ausubel et al. (above). A phage encoding a variable region with the desired specificity will then transmit the desired antigen (i.e., Selected for specific binding to hK2, including the selected variable domain, either whole or partially. A specific antibody is reconstituted. The nucleic acid sequence encoding the reconstituted antibody is produced by the hybridoma. It is introduced into suitable cell lines such as myeloma cells used in live production, and as a result, monoclonal anti Antibodies with specific characteristics of the body are secreted by cells (for example, Janeway et al.) (Above), Huse et al. (Above), and U.S. Patent No. 6,265,150 (See reference).

[0132] Antibodies are transgenic of specific heavy and light chain immunoglobulin genes. It can be produced by a sgenic mouse. Such a method is already in use in the art. This is knowledge, for example, U.S. Patent No. 5,545,806 and No. 5,569,825, and Janeway et al., as listed above.

[0133] Methods for producing humanized antibodies are known in the art, for example, Janew ay et al., U.S. Patent No. 5,225,539, No. 5,585,089, and No. Patent No. 5,693,761, European Patent No. 0239400(B1), and British Patent No. 218 It is described in U.S. Patent No. 8638. Humanized antibodies are also described in U.S. Patent No. 5,639,641 and Pedersen et al., J. Mol. Biol., 235, 959-973( It can also be produced using the antibody resurfacing technique described in 1994.

[0134] When used herein, antibodies may be multiple or single chains, or intact immunoglobulins. The antibody may be immunoglobulin, and may be derived from natural or recombinant sources. It can be a tetramer of phosphorus molecules.

[0135] The term "antibody fragment" refers to an intact antibody that retains the antigen-binding properties of the parent's full-length antibody. This refers to at least one part of the recombinant variant. This could be, for example, recognition and conclusion. Sufficient to confer binding (for example, specific binding of an antibody fragment to a target such as an antigen) This refers to the antigen-binding domain (for example, the antigen-determining variable region of an intact antibody). An "antigen-binding fragment" refers to a part of an immunoglobulin molecule. An example of an antibody fragment is: Fab, Fab', F(ab')2, and Fv fragments, single-chain antibody (scFv), linear antibody , single-domain antibodies such as sdAb (either VL or VH), camel VHH domain Examples include, but are not limited to, multispecific antibodies formed from antibody fragments. .

[0136] The term "scFv" refers to an antibody fragment containing at least one variable region of the light chain and a heavy chain. This refers to a protein containing at least one antibody fragment that includes a variable region of . In the application configuration, the light chain variable region and the heavy chain variable region are connected via short, flexible polypeptide linkers. They are linked together in a continuous chain and can be expressed as a single-chain polypeptide, and scFv is its original The specificity of the intact antibody is preserved. Unless otherwise specified, when used herein, scFv may have the VL and VH variable regions in any order, for example, polyp Regarding the N-terminus and C-terminus of the cyd, scFv may include VL-linker-VH. It may also contain VH-linker-VL.

[0137] One embodiment of the present invention also provides an antigen-binding moiety of any of the antibodies described herein. The antigen-binding portion consists of Fab, F(ab')2, dsFv, sFv, diabody, and It may be any part having at least one antigen-binding site, such as a triabody.

[0138] In some embodiments, the antigen-binding fragment is a heavy chain complementarity determining region. Mentarity determining regions (HCDR) 1, 2, and / or 3, light chain complementarity determination Light chain complementarity determining regions (LCDR) 1, 2, and / or 3, heavy chain variable region (VH), or light chain variable region (VL), Fab, F(ab' )2, Fd, and Fv fragments, and either one VH domain or one VL domain It is a domain antibody (dAb) containing (for example, consisting of) the VH domain and the VL domain. The main components can be linked together via a linker, such as a synthetic linker.

[0139] "Complementarity determining regions (CDRs)" are found in antibodies. It is an antigen-binding site. CDR can be defined using various terms: (i) within VH There are three in (HCDR1, HCDR2, HCDR3) and three in VL (LCDR1, LCD The complementarity-determining region (CDR) of R2 (LCDR3) is based on sequence diversity (Wu and Kabat,J Exp Med 132:211-50,1970;Kaba t et al.,Sequences of Proteins of Immuno logical Interest,5th Ed.Public Health Se rvice,National Institutes of Health,Beth (esda, Md., 1991). (ii) Three within VH (H1, H2, H3) and within VL The three (L1, L2, L3) "super variable regions", "HVR", or "HV" are Cho thia and Lesk(Chothia and Lesk Mol Biol 196 As defined by (901-17, 1987), it is highly variable in structure, anti This refers to the region of the body variable domain. The cs(IMGT) database (http: / / www_imgt_org) is an antigenic database. Provides standard numbers and definitions for the combined parts. Summary of CDR, HV, and IMGT. The response to this is from Lefranc et al., Dev Comparat Immunol This is described in 27:55-77, 2003. The term "CDR" as used in this specification. , "HCDR1", "HCDR2", "HCDR3", "LCDR1", "LCDR2" The terms "LCDR3" and "LCDR3" refer to the above-mentioned Ka unless otherwise specified herein. Defined by any of the methods described by bat, Chothia, or IMGT. Includes the CDR.

[0140] Furthermore, the antibody or its antigen-binding portion may be, for example, a radioactive isotope, a fluorophore (for example) Fluorescein isothiocyanate (FITC), Coerythrin (phycoerythrin, PE), enzymes (e.g., alkaline phosphatase, West) A detectable label such as horseradish peroxidase and elemental particles (e.g., gold particles) It can be modified to include.

[0141] CARs, polypeptides, or proteins (functional portions and their functional components) as described herein A nucleic acid comprising a nucleotide sequence encoding any of the variants is described in this disclosure. It is provided as such.

[0142] A portion of the CAR containing an antibody or an antibody fragment may exist in various forms, in which case The antigen-binding domain is, for example, a single-domain antibody fragment (sdAb), scFv, and human It is expressed as part of a continuous polypeptide chain such as a chimeric antibody or a humanized antibody (Ha rlow et al., 1999, In: Using Antibodies:AL aboratory Manual,Cold Spring Harbor Labo ratory Press, NY; Harlow et al., 1989, In: Antibodies:A Laboratory Manual,Cold Spri ng Harbor, NY; Houston et al., 1988, Proc. Natl.Acad.Sci.USA 85:5879-5883;Bird et a l., 1988, Science 242:423-426). In one embodiment, C of the present invention The antigen-binding domain of the AR composition contains an antibody fragment. In one embodiment, the CAR contains scFv Contains antibody fragments.

[0143] The term "recombinant antibody" is used, for example, by bacteriophage or yeast expression systems. This refers to antibodies produced using recombinant DNA technology, such as expressed antibodies. These DNA molecules, which are generated by the synthesis of DNA molecules that code for the body, are antibody proteins. It should be interpreted as meaning an antibody that expresses an amino acid sequence that identifies the antibody. In that case, the DNA or amino acid sequence is available and known in the art. It is obtained using a recombinant DNA or amino acid sequencing technique.

[0144] The term "antigen" refers to a molecule that triggers an immune response. This immune response involves antibody production. This may involve the activation of specific immunocompetent cells, or both. This means that virtually any macromolecule, including all proteins or peptides, can function as an antigen. They will understand that. Furthermore, antigens can be recombinant or derived from genomic DNA. If you are a professional, any DNA can code for a nucleotype that generates a protein that triggers an immune response. The term is used herein by including a nucleotide sequence or a partial nucleotide sequence. They will understand that the antigen codes as such. Furthermore, those skilled in the art will understand that the antigen is It is important to understand that genes do not need to be encoded solely by their full-length nucleotide sequence. This disclosure involves the use of partial nucleotide sequences of two or more genes, but this is not applicable. Not limited to, and these nucleotide sequences are arranged in various combinations. It is clear that this encodes polypeptides that induce the desired immune response.

[0145] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and the extracellular antigen-binding domain is transmitted to the hK2 antigen. Combine.

[0146] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 1 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 4 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 2 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 4 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 3 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 6 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 3 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 4 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 2 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 6 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 1 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 6 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 3 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 5 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 1 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 5 CDR3; Light chain CDR1, CDR2, and CDR from the variable region containing the amino acid sequence of SEQ ID NO: 2 3, and heavy chain CDR1, CDR2, and from the variable region including the amino acid sequence of SEQ ID NO: 5 CDR3; Light chain CDR1, CDR2, and CD from the variable region containing the amino acid sequence of SEQ ID NO: 74 Heavy chain CDR1 and CDR2 from the variable region including R3 and the amino acid sequence of SEQ ID NO: 76. , and CDR3; Light chain CDR1, CDR2, and CD from the variable region containing the amino acid sequence of SEQ ID NO: 75 Heavy chain CDR1 and CDR2 from the variable region including R3 and the amino acid sequence of SEQ ID NO: 77 , and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 147 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 140. R2 and CDR3; Light chain CDR1, CDR2, and CD from the variable region containing the amino acid sequence of SEQ ID NO: 75 Heavy chain CDR1 and CDR from the variable region including R3 and the amino acid sequence of SEQ ID NO: 141. 2, and CDR3; Light chain CDR1, CDR2, and CD from the variable region containing the amino acid sequence of SEQ ID NO: 75 Heavy chain CDR1 and CDR from the variable region including R3 and the amino acid sequence of SEQ ID NO: 140. 2, and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 148 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 142. R2 and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 149 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 143. R2 and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 150 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 144. R2 and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 151 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 145. R2 and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 152 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 146. R2 and CDR3; Light chain CDR1, CDR2, and C from the variable region containing the amino acid sequence of SEQ ID NO: 318 Heavy chain CDR1 and CD from the variable region including DR3 and the amino acid sequence of SEQ ID NO: 317. R2 and CDR3 Includes, The extracellular antigen-binding domain binds to the hK2 antigen.

[0147] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 1. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 4. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0148] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 2. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 4. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0149] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 3. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 6. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0150] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 3. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 4. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0151] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 2. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 6. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0152] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 1. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 6. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0153] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 3. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 5. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0154] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 1. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 5. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0155] In one embodiment, the extracellular antigen-binding domain is a variable region containing the amino acid sequence of SEQ ID NO: 2. Includes the light chains CDR1, CDR2, and CDR3 from the region, as well as the amino acid sequence of SEQ ID NO: 5. It includes heavy chains CDR1, CDR2, and CDR3 from the variable region.

[0156] In one embodiment, the extracellular antigen-binding domain is variable, containing the amino acid sequence of SEQ ID NO: 74. Light chain CDR1, CDR2, and CDR3 from the region, and amino acid sequence of SEQ ID NO: 76 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region.

[0157] In one embodiment, the extracellular antigen-binding domain is variable, containing the amino acid sequence of SEQ ID NO: 75. Light chain CDR1, CDR2, and CDR3 from the region, and amino acid sequence of SEQ ID NO: 77 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region.

[0158] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 147. Light chain CDR1, CDR2, and CDR3 from the mutational region, and amino acids of SEQ ID NO: 140 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0159] In one embodiment, the extracellular antigen-binding domain is variable, containing the amino acid sequence of SEQ ID NO: 75. Light chains CDR1, CDR2, and CDR3 from the region, and amino acid sequence of SEQ ID NO: 141 Includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing columns.

[0160] In one embodiment, the extracellular antigen-binding domain is variable, containing the amino acid sequence of SEQ ID NO: 75. Light chains CDR1, CDR2, and CDR3 from the region, and amino acid sequence of SEQ ID NO: 140 Includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing columns.

[0161] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 148. Light chain CDR1, CDR2, and CDR3 from the mutation region, and amino acids of SEQ ID NO: 142 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0162] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 149. Light chain CDR1, CDR2, and CDR3 from the mutation region, and amino acids of SEQ ID NO: 143 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0163] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 150. Light chain CDR1, CDR2, and CDR3 from the mutation region, and amino acids of SEQ ID NO: 144 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0164] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 151. Light chain CDR1, CDR2, and CDR3 from the mutation region, and amino acids of SEQ ID NO: 145 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0165] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 152. Light chain CDR1, CDR2, and CDR3 from the mutation region, and amino acids of SEQ ID NO: 146 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0166] In one embodiment, the extracellular antigen-binding domain may include the amino acid sequence of SEQ ID NO: 318. Light chain CDR1, CDR2, and CDR3 from the mutation region, and amino acids of SEQ ID NO: 317 It includes heavy chains CDR1, CDR2, and CDR3 from a variable region containing sequences.

[0167] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Sequence ID 63, Sequence ID 72, Sequence ID 86, Sequence ID 92, Sequence ID 102, Sequence Number Number 105, Sequence ID 107, Sequence ID 110, Sequence ID 115, Sequence ID 118, Sequence Number Sequence ID 120, Sequence ID 123, Sequence ID 309, and Sequence ID 314, and their storage The extracellular environment contains a heavy chain CDR1 having an amino acid sequence selected from the group consisting of specific modifications. The antigen-binding domain binds to the hK2 antigen.

[0168] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 73, SEQ ID NO: 87, SEQ ID NO: 93, SEQ ID NO: 103, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 119, SEQ ID NO: 121, SEQ ID NO: 124, SEQ ID NO: Ami selected from the group consisting of 310, SEQ ID NO: 315, and their conservative modifications. It contains a heavy chain CDR2 having a noacid sequence, and the extracellular antigen-binding domain binds to the hK2 antigen. To combine.

[0169] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Sequence ID 66, Sequence ID 88, Sequence ID 94, Sequence ID 104, Sequence ID 109, Array Number 113, Sequence ID 117, Sequence ID 122, and Sequence ID 311, and their respective reserves The cell contains a heavy chain CDR3 having an amino acid sequence selected from the group consisting of residual modifications. The external antigen-binding domain binds to the hK2 antigen.

[0170] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 4, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, SEQ ID NO: A heavy chain CDR2 having a 315-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 311. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 309, SEQ ID NO: A heavy chain CDR2 having a 310-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 311. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having the amino acid sequence of 3, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 86, SEQ ID NO: 8 Heavy chain CDR2 having the amino acid sequence of 7, and heavy chain having the amino acid sequence of SEQ ID NO: 88 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 92, SEQ ID NO: 9 Heavy chain CDR2 having amino acid sequence 3, and heavy chain having amino acid sequence of SEQ ID NO: 94 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: A heavy chain CDR2 having the amino acid sequence of 103, and having the amino acid sequence of SEQ ID NO: 104 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: A heavy chain CDR2 having a sequence of 106 amino acids, and having the amino acid sequence of SEQ ID NO: 104 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: A heavy chain CDR2 having a sequence of 108 amino acids, and having the amino acid sequence of SEQ ID NO: 109 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 110, SEQ ID NO: A heavy chain CDR2 having an amino acid sequence of 111, and having the amino acid sequence of SEQ ID NO: 109 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: A heavy chain CDR2 having a 112-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 113. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: A heavy chain CDR2 having a sequence of 114 amino acids, and having the amino acid sequence of SEQ ID NO: 113. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: A heavy chain CDR2 having a sequence of 116 amino acids, and having the amino acid sequence of SEQ ID NO: 117 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: A heavy chain CDR2 having a sequence of 119 amino acids, and having the amino acid sequence of SEQ ID NO: 117 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: A heavy chain CDR2 having a 121-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 122. heavy chain CDR3; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 123, SEQ ID NO: A heavy chain CDR2 having a 124-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 122. heavy chain CDR3, Includes, The extracellular antigen-binding domain binds to the hK2 antigen.

[0171] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 63. Chain complementarity determination region 1 (CDR1), heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 64, It also contains a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 66.

[0172] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 314 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 315 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 311.

[0173] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 309. Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 310 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 311.

[0174] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 63. Chain complementarity determination region 1 (CDR1), heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 65, It also contains a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 66.

[0175] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 72 Chain complementarity determination region 1 (CDR1), heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 73, It also contains a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 66.

[0176] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 86. Chain complementarity determination region 1 (CDR1), heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 87, It also contains a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 88.

[0177] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 92 Chain complementarity determination region 1 (CDR1), heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 93, It also contains a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 94.

[0178] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 102 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 103 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 104.

[0179] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 105 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 106 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 104.

[0180] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 107 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 108 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 109.

[0181] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 110 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 111 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 109.

[0182] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 102 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 112 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 113.

[0183] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 105 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 114 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 113.

[0184] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 115 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 116 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 117.

[0185] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 118 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 119 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 117.

[0186] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 120 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 121 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 122.

[0187] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 123 Heavy chain CDR having heavy chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 124 It contains 2 and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 122.

[0188] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 89, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 125, SEQ ID NO: 128, SEQ ID NO: 133, SEQ ID NO: 136, and SEQ ID NO: 312, and It contains a light chain CDR1 having an amino acid sequence selected from the group consisting of these conserved modifications. Furthermore, the extracellular antigen-binding domain binds to the hK2 antigen.

[0189] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Sequence ID 69, Sequence ID 70, Sequence ID 90, Sequence ID 96, Sequence ID 126, Sequence Number Sequence No. 137, Sequence No. 129, Sequence No. 131, Sequence No. 134, Sequence No. 137, and The amino acid sequence having sequence number 313, and selected from the group consisting of these conserved modifications. It contains a light chain CDR2, and the extracellular antigen-binding domain binds to the hK2 antigen.

[0190] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Sequence ID 71, Sequence ID 91, Sequence ID 130, Sequence ID 132, Sequence ID 135, Distribution Selected from the group consisting of column number 138, sequence number 139, and their conservative modifications. It contains a light chain CDR3 having an amino acid sequence, and the extracellular antigen-binding domain is an hK2 anti- Combine with the original.

[0191] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 6 Light chain CDR2 having the amino acid sequence of 9, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, SEQ ID NO: 7 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 95, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence of 6, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 312, SEQ ID NO: A light chain CDR2 having a 313-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 91 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having amino acid sequence 0, and light chain having amino acid sequence SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 1 A light chain CDR2 having a 26-amino acid sequence, and a light chain having the amino acid sequence of SEQ ID NO: 71. chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 128, SEQ ID NO: A light chain CDR2 having a 129-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 130. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 125, SEQ ID NO: A light chain CDR2 having a 131-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 132. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 133, SEQ ID NO: A light chain CDR2 having a 134-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 135. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 136, SEQ ID NO: A light chain CDR2 having a sequence of 137 amino acids, and having the amino acid sequence of SEQ ID NO: 138. Light chain CDR3; or Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 97, SEQ ID NO: 1 A light chain CDR2 having a 27-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 139 Light chain CDR3, Includes, The extracellular antigen-binding domain binds to the hK2 antigen.

[0192] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 67. Chain complementarity determination region 1 (CDR1), light chain CDR2 having the amino acid sequence of SEQ ID NO: 69, It also contains a light chain CDR3 having the amino acid sequence of SEQ ID NO: 71.

[0193] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 68. Chain complementarity determination region 1 (CDR1), light chain CDR2 having the amino acid sequence of SEQ ID NO: 70, It also contains a light chain CDR3 having the amino acid sequence of SEQ ID NO: 71.

[0194] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 89. Chain complementarity determination region 1 (CDR1), light chain CDR2 having the amino acid sequence of SEQ ID NO: 90, It also contains a light chain CDR3 having the amino acid sequence of SEQ ID NO: 91.

[0195] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 95. Chain complementarity determination region 1 (CDR1), light chain CDR2 having the amino acid sequence of SEQ ID NO: 96, It also contains a light chain CDR3 having the amino acid sequence of SEQ ID NO: 71.

[0196] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 312 Light chain complementarity determination region 1 (CDR1), light chain CDR having the amino acid sequence of SEQ ID NO: 313 It contains a light chain CDR3 having the amino acid sequence of 2 and SEQ ID NO: 91.

[0197] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 89. Chain complementarity determination region 1 (CDR1), light chain CDR2 having the amino acid sequence of SEQ ID NO: 90, It also contains a light chain CDR3 having the amino acid sequence of SEQ ID NO: 91.

[0198] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 67. Light chain CDR2 having chain complementarity determination region 1 (CDR1) and the amino acid sequence of sequence number 126. , and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 71.

[0199] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 128 Light chain CDR having light chain complementarity determination region 1 (CDR1) and the amino acid sequence of sequence number 129 It contains a light chain CDR3 having the amino acid sequence of 2 and SEQ ID NO: 130.

[0200] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 125 Light chain CDR1 having the amino acid sequence of SEQ ID NO: 131 It contains a light chain CDR3 having the amino acid sequence of 2 and SEQ ID NO: 132.

[0201] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 133 Light chain CDR having light chain complementarity determination region 1 (CDR1) and the amino acid sequence of SEQ ID NO: 134 It contains a light chain CDR3 having the amino acid sequence of 2 and SEQ ID NO: 135.

[0202] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 136 Light chain CDR1, having the amino acid sequence of SEQ ID NO: 137 It contains a light chain CDR3 having the amino acid sequence of 2 and SEQ ID NO: 138.

[0203] In one embodiment, the extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 97. Light chain CDR2 having chain complementarity determination region 1 (CDR1) and the amino acid sequence of sequence number 127. , and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 139.

[0204] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 4, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, Light chain CDR2 having the amino acid sequence of sequence number 70, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 315, heavy chain having an amino acid sequence of SEQ ID NO: 311 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 312 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 313, amino acid sequence of SEQ ID NO: 91 Light chain CDR3 having a row; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having amino acid sequence 103, heavy chain having amino acid sequence of SEQ ID NO: 104 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 128 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 129, amino acids of SEQ ID NO: 130 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: Heavy chain CDR2 having a 108 amino acid sequence, heavy chain having the amino acid sequence of SEQ ID NO: 109 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 125 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 131, amino acids of SEQ ID NO: 132 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 112, heavy chain having an amino acid sequence of SEQ ID NO: 113 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and SEQ ID NO: 133 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 134, amino acids of SEQ ID NO: 135 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 116, heavy chain having an amino acid sequence of SEQ ID NO: 117 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of sequence number 136 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 137, amino acids of SEQ ID NO: 138 Light chain CDR3 having a sequence; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 121, heavy chain having an amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 138 Light chain CDR3 having a row, Includes, The extracellular antigen-binding domain binds to the hK2 antigen.

[0205] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 4, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, Light chain CDR2 having the amino acid sequence of sequence number 70, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 315, heavy chain having an amino acid sequence of SEQ ID NO: 311 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 312 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 313, amino acid sequence of SEQ ID NO: 91 Light chain CDR3 having a row; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having amino acid sequence 103, heavy chain having amino acid sequence of SEQ ID NO: 104 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 128 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 129, amino acids of SEQ ID NO: 130 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: Heavy chain CDR2 having a 108 amino acid sequence, heavy chain having the amino acid sequence of SEQ ID NO: 109 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 125 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 131, amino acids of SEQ ID NO: 132 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 112, heavy chain having an amino acid sequence of SEQ ID NO: 113 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and SEQ ID NO: 133 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 134, amino acids of SEQ ID NO: 135 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 116, heavy chain having an amino acid sequence of SEQ ID NO: 117 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of sequence number 136 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 137, amino acids of SEQ ID NO: 138 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 121, heavy chain having an amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 138 Light chain CDR3 having a row; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, Light chain CDR2 having the amino acid sequence of sequence number 70, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 309, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 310, heavy chain having an amino acid sequence of SEQ ID NO: 311 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 312 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 313, amino acid sequence of SEQ ID NO: 91 Light chain CDR3 having a row; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 106, heavy chain having an amino acid sequence of SEQ ID NO: 104 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 128 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 129, amino acids of SEQ ID NO: 130 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 110, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 111, heavy chain having an amino acid sequence of SEQ ID NO: 109 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 125 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 131, amino acids of SEQ ID NO: 132 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 114, heavy chain having an amino acid sequence of SEQ ID NO: 113 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and SEQ ID NO: 133 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 134, amino acids of SEQ ID NO: 135 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 119, heavy chain having an amino acid sequence of SEQ ID NO: 117 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of sequence number 136 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 137, amino acids of SEQ ID NO: 138 Light chain CDR3 having a sequence; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 123, SEQ ID NO: Heavy chain CDR2 having a 124-amino acid sequence, heavy chain having the amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 139 Light chain CDR3 having a row; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 121, heavy chain having an amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 139 Light chain CDR3 having a column; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 86, SEQ ID NO: 8 Heavy chain CDR2 has the amino acid sequence of 7, and heavy chain CD has the amino acid sequence of SEQ ID NO: 88. R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, Light chain CDR2 having the amino acid sequence of sequence number 90, and having the amino acid sequence of sequence number 91 Light chain CDR3, Includes, The extracellular antigen-binding domain binds to the hK2 antigen.

[0206] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 4, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Contains light chain CDR3.

[0207] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, Light chain CDR2 having the amino acid sequence of sequence number 70, and having the amino acid sequence of sequence number 71 Contains light chain CDR3.

[0208] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. It contains light chain CDR3.

[0209] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Contains light chain CDR3.

[0210] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 with amino acid sequence 5, heavy chain CD with amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. It contains light chain CDR3.

[0211] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 315, heavy chain having an amino acid sequence of SEQ ID NO: 311 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 312 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 313, amino acid sequence of SEQ ID NO: 91 It contains a light chain CDR3 having a column.

[0212] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having amino acid sequence 103, heavy chain having amino acid sequence of SEQ ID NO: 104 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 128 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 129, amino acids of SEQ ID NO: 130 It contains a light chain CDR3 having a sequence.

[0213] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: Heavy chain CDR2 having a 108 amino acid sequence, heavy chain having the amino acid sequence of SEQ ID NO: 109 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 125 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 131, amino acids of SEQ ID NO: 132 It contains a light chain CDR3 having a sequence.

[0214] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 112, heavy chain having an amino acid sequence of SEQ ID NO: 113 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and SEQ ID NO: 133 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 134, amino acids of SEQ ID NO: 135 It contains a light chain CDR3 having a sequence.

[0215] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 116, heavy chain having an amino acid sequence of SEQ ID NO: 117 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of sequence number 136 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 137, amino acids of SEQ ID NO: 138 It contains a light chain CDR3 having a sequence.

[0216] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 121, heavy chain having an amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 138 It contains a light chain CDR3 having a column.

[0217] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Contains light chain CDR3.

[0218] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, Light chain CDR2 having the amino acid sequence of sequence number 70, and having the amino acid sequence of sequence number 71 Contains light chain CDR3.

[0219] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. It contains light chain CDR3.

[0220] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Contains light chain CDR3.

[0221] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having amino acid sequence 3, heavy chain CD having amino acid sequence of SEQ ID NO: 66 R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, The light chain CDR2 having the amino acid sequence of sequence number 126, and the amino acid sequence of sequence number 71 It contains the light chain CDR3.

[0222] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 309, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 310, heavy chain having an amino acid sequence of SEQ ID NO: 311 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 312 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 313, amino acid sequence of SEQ ID NO: 91 It contains a light chain CDR3 having a column.

[0223] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 106, heavy chain having an amino acid sequence of SEQ ID NO: 104 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 128 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 129, amino acids of SEQ ID NO: 130 It contains a light chain CDR3 having a sequence.

[0224] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 110, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 111, heavy chain having an amino acid sequence of SEQ ID NO: 109 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and sequence number 125 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 131, amino acids of SEQ ID NO: 132 It contains a light chain CDR3 having a sequence.

[0225] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 114, heavy chain having an amino acid sequence of SEQ ID NO: 113 Light chain complementarity determination region 1 (CDR) having the amino acid sequence of chain CDR3 and SEQ ID NO: 133 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 134, amino acids of SEQ ID NO: 135 It contains a light chain CDR3 having a sequence.

[0226] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 119, heavy chain having an amino acid sequence of SEQ ID NO: 117 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of sequence number 136 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 137, amino acids of SEQ ID NO: 138 It contains a light chain CDR3 having a sequence.

[0227] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 123, SEQ ID NO: Heavy chain CDR2 having a 124-amino acid sequence, heavy chain having the amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 139 It contains a light chain CDR3 having a column.

[0228] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 121, heavy chain having an amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and sequence number 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 139 It contains a light chain CDR3 having a column.

[0229] In one embodiment, the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 86, SEQ ID NO: 8 Heavy chain CDR2 has the amino acid sequence of 7, and heavy chain CD has the amino acid sequence of SEQ ID NO: 88. R3, and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, Light chain CDR2 having the amino acid sequence of sequence number 90, and having the amino acid sequence of sequence number 91 Contains light chain CDR3.

[0230] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. The CAR provides a transmission domain, and this extracellular antigen-binding domain is Light chain variable region (LCVR) containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-3 ), or a heavy chain variable region (H) containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 4-6. CVR), or LCVR containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-3 A combination of HCVR containing an amino acid sequence selected from the group consisting of SEQ ID NOs: 4-6 The extracellular antigen-binding domain contains and binds to the hK2 antigen.

[0231] In one embodiment, the extracellular antigen-binding domain is Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; The light chain variable region includes the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of SEQ ID NO: 76. heavy chain variable region; light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 77 Heavy chain variable region containing a mino acid sequence; Light chain variable region containing the amino acid sequence of SEQ ID NO: 147 and amino acid sequence of SEQ ID NO: 140 Heavy chain variable region including; The light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 141 Includes heavy chain variable region; The light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 140 Includes heavy chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 148 and amino acid sequence of SEQ ID NO: 142 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 149 and amino acid sequence of SEQ ID NO: 143 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 150 and amino acid sequence of SEQ ID NO: 144 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 151 and amino acid sequence of SEQ ID NO: 145 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 152 and amino acid sequence of SEQ ID NO: 146 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 318 and amino acid sequence of SEQ ID NO: 317 Heavy chain variable region including, Includes.

[0232] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 1. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4.

[0233] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 2. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4.

[0234] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 3. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6.

[0235] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 3. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4.

[0236] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 2. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6.

[0237] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 1. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6.

[0238] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 1. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5.

[0239] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 2. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5.

[0240] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 3. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5.

[0241] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 74. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 76.

[0242] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 75. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 77.

[0243] In one embodiment, the extracellular antigen-binding domain contains the amino acid sequence of SEQ ID NO: 147. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 140.

[0244] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 75. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 141.

[0245] In one embodiment, the extracellular antigen-binding domain is a light chain containing the amino acid sequence of SEQ ID NO: 75. It includes a variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 140.

[0246] In one embodiment, the extracellular antigen-binding domain contains the amino acid sequence of SEQ ID NO: 148. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 142.

[0247] In one embodiment, the extracellular antigen-binding domain contains the amino acid sequence of SEQ ID NO: 149. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 143.

[0248] In one embodiment, the extracellular antigen-binding domain includes the amino acid sequence of SEQ ID NO: 150. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 144.

[0249] In one embodiment, the extracellular antigen-binding domain includes the amino acid sequence of SEQ ID NO: 151. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 145.

[0250] In one embodiment, the extracellular antigen-binding domain contains the amino acid sequence of SEQ ID NO: 152. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 146.

[0251] In one embodiment, the extracellular antigen-binding domain contains the amino acid sequence of SEQ ID NO: 318. It includes a chain variable region and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 317.

[0252] In one embodiment, the extracellular antigen-binding domain is Sequence ID 1 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 4 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence ID 2 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 4 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence number 3 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 6 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence number 3 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 4 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence ID 2 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 6 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence number 3 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 4 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence ID 1 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 6 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence number 3 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 5 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence ID 1 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 5 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence ID 2 and at least 50, at least 55, at least 60, at least 65, small At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence, and at least SEQ ID NOs. 5 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A heavy chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity; Sequence ID 74 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having, and at least 50, and at least 76 and 50 At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Heavy chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity area; Number 75 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having such a sequence, and at least 50, and at least 50 of sequence numbers 77 and 50. At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Heavy chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity area; Sequence ID 147 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having a property, and at least sequence numbers 140 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain; Number 75 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having such a sequence, and at least 50 of sequence numbers 141 and 50. At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain variable containing an amino acid sequence with at least 98, or at least 99%, sequence identity. region; Number 75 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having, and at least SEQ ID NOs. 140 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain variable containing an amino acid sequence with at least 98, or at least 99%, sequence identity. region; Sequence ID 148 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having a property, and at least SEQ ID NOs. 142 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain; Sequence ID 149 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having a property, and at least SEQ ID NOs. 143 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain; Sequence ID 150 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having a property, and at least SEQ ID NOs. 144 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain; Sequence ID 151 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having a property, and at least SEQ ID NOs. 145 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain; Sequence ID 152 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having a property, and at least SEQ ID NOs. 146 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain; Sequence ID 318 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 A light chain variable region containing an amino acid sequence having properties, and at least 50 of sequence numbers 317 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, Heavy chain containing amino acid sequences with at least 98, or at least 99%, sequence identity. Variable domain Includes.

[0253] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 1 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence ID No. 4 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0254] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 2 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence ID No. 4 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0255] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 3 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence No. 6 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0256] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 3 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence ID No. 4 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0257] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 2 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence No. 6 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0258] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 3 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence ID No. 4 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0259] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 1 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence No. 6 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0260] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 3 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence No. 5 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0261] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 1 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence No. 5 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0262] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 2 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, and at least 80, at least 85, at least 90, at least 91, at least 92, and at least 93, at least 94, at least 95, at least 96, at least 97, and at least A light chain variable region containing an amino acid sequence having 98, or at least 99%, sequence identity, Also, Sequence No. 5 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, 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 It contains a heavy chain variable region that includes an amino acid sequence having properties.

[0263] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 74 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, less 80, at least 85, at least 90, at least 91, at least 92, less 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences having 98% or at least 99% sequence identity. , and also sequence number 76 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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% of the array It contains a heavy chain variable region that includes an identical amino acid sequence.

[0264] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 75 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, less 80, at least 85, at least 90, at least 91, at least 92, less 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences having 98% or at least 99% sequence identity. , as well as sequence number 77 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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% of the array It contains a heavy chain variable region that includes an identical amino acid sequence.

[0265] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 147 and 50, at least At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 140 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0266] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 75 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, less 80, at least 85, at least 90, at least 91, at least 92, less 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences having 98% or at least 99% sequence identity. , and sequence number 141 and at least 50, at least 55, at least 60, and 65, at least 70, at least 75, at least 80, at least 85, and at least 90, at least 91, at least 92, at least 93, at least 94, and at least 95, at least 96, at least 97, at least 98, or at least 99% It contains a heavy chain variable region that includes an amino acid sequence having column identity.

[0267] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 75 and 50, and at least 55, at least 60, at least 65, at least 70, at least 75, less 80, at least 85, at least 90, at least 91, at least 92, less 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences having 98% or at least 99% sequence identity. , as well as sequence number 140 and at least 50, at least 55, at least 60, and 65, at least 70, at least 75, at least 80, at least 85, and at least 90, at least 91, at least 92, at least 93, at least 94, and at least 95, at least 96, at least 97, at least 98, or at least 99% It contains a heavy chain variable region that includes an amino acid sequence having column identity.

[0268] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 148 and 50, at least At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 142 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0269] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 149 and 50, and at least At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 143 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0270] In one embodiment, the extracellular antigen-binding domain is at least 50 of sequence number 150, and at least 50 At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 144 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0271] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 151 and 50, at least At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 145 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0272] In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 152 and at least 50. At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 146 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0273] In one embodiment, the extracellular antigen-binding domain is at least 50 of sequence number 318, and at least 50 At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least Light chain variable regions containing amino acid sequences with at least 98, or at least 99%, sequence identity The region, and sequence number 317 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95%, at least 96%, at least 97%, at least 98%, or at least 99% It includes a heavy chain variable region containing an amino acid sequence with sequence identity.

[0274] In one embodiment, the extracellular antigen-binding domain includes scFv. In several embodiments, scFv contains a linker polypeptide between the light chain variable region and the heavy chain variable region. In a specific embodiment, the extracellular antigen-binding domain is sequence numbers 8-23, 169-184, and The hK2 polypeptide contains an amino acid sequence selected from the group consisting of 340-343. For example, a human hK2 polypeptide or a fragment thereof having the amino acid sequence of SEQ ID NO: 62 This is a scFv that binds specifically.

[0275] In recombinant expression systems, the linker is a peptide linker, which is present in any naturally occurring environment. It may contain amino acids. Examples of amino acids that may be present in the linker are Gly, Ser, and Pr. These are o, Thr, Glu, Lys, Arg, Ile, Leu, His, and The. The Kerrs maintain a precise higher-order configuration relative to each other to preserve desired activities, such as binding to hK2. It is necessary to have an appropriate length to connect VH and VL in a way that forms a structure. be.

[0276] The linker can be approximately 5 to 50 amino acids long. In some embodiments, the linker - has an amino acid length of approximately 10 to 40. In some embodiments, the linker is approximately 1 The amino acid length is 0 to 35. In some embodiments, the linker is approximately 10 to 30. This is the length of the amino acids. In some embodiments, the linker is about 10 to 25 amino acids. It is long. In some embodiments, the linker is about 10 to 20 amino acids long. In some embodiments, the linker is about 15-20 amino acids long. In some embodiments, the linker has a length of 6 amino acids. It has a length of 7 amino acids. In some embodiments, the linker has a length of 8 amino acids. In some embodiments, the linker is 9 amino acid length. The linker is 10 amino acids long. In some embodiments, the linker is 11 The length is one amino acid. In some embodiments, the linker has a length of 12 amino acids. In some embodiments, the linker has a length of 13 amino acids. The linker has a length of 14 amino acids. In some embodiments, the linker has a length of 15 amino acids. The length is one amino acid. In some embodiments, the linker has a length of 16 amino acids. In some embodiments, the linker has a length of 17 amino acids. The linker has a length of 18 amino acids. In some embodiments, the linker has a length of 19 amino acids. The length is 1 amino acid. In some embodiments, the linker is 20 amino acid length. In some embodiments, the linker has a length of 21 amino acids. The linker has a length of 22 amino acids. In some embodiments, the linker has a length of 23 amino acids. The length is one amino acid. In some embodiments, the linker has a length of 24 amino acids. In some embodiments, the linker has a length of 25 amino acids. The linker has a length of 26 amino acids. In some embodiments, the linker is 27 The length is one amino acid. In some embodiments, the linker has a length of 28 amino acids. In some embodiments, the linker has a length of 29 amino acids. The linker is 30 amino acids long. In some embodiments, the linker is 31 The length is one amino acid. In some embodiments, the linker has a length of 32 amino acids. In some embodiments, the linker has a length of 33 amino acids. The linker has a length of 34 amino acids. In some embodiments, the linker has a length of 35 amino acids. The length is one amino acid. In some embodiments, the linker has a length of 36 amino acids. In some embodiments, the linker has a length of 37 amino acids. The linker has a length of 38 amino acids. In some embodiments, the linker has a length of 39 amino acids. The length is 1 amino acid. In some embodiments, the linker is 40 amino acid length. . Linkers containing carbides, glycerides and ala, linkers containing ala and ser, and other flexible linkers. It's a linker.

[0277] In one embodiment, the linker polypeptide contains the amino acid sequence of SEQ ID NO: 7. Morphologically, the linker polypeptide is at least SEQ ID NO: 7 and at least 50, at least 55, At least 60, at least 65, at least 70, at least 75, 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, and It contains an amino acid sequence having at least 99% sequence identity. In one embodiment, a linker - The polypeptide contains one of the amino acid sequences from SEQ ID NOs. 237-268. In the application form, the linker polypeptide is one of the sequence numbers 237-268 and a small number of others. 50, at least 55, at least 60, at least 65, at least 70, less 75, at least 80, at least 85, at least 90, at least 91, less 92, at least 93, at least 94, at least 95, at least 96, less Both amino acid sequences have 97%, at least 98%, or at least 99% sequence identity. include.

[0278] In one embodiment, scFv is sequence numbers 8-23, 169-184, and 340-34 It comprises an amino acid sequence selected from the group consisting of 3. In one embodiment, scFv is the sequence number Number 8 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 Having 96, at least 97, at least 98, or at least 99% sequence identity It includes an amino acid sequence or the sequence of SEQ ID NO: 8. In one embodiment, scFv is SEQ ID NO: 9 and at least 50, at least 55, at least 60, at least 65, at least 7 0, at least 75, at least 80, at least 85, at least 90, at least 9 1, at least 92, at least 93, at least 94, at least 95, at least 9 6. Having at least 97%, at least 98%, or at least 99% sequence identity It includes an amino acid sequence or the sequence of SEQ ID NO: 9. In one embodiment, scFv is SEQ ID NO: 1 0 and at least 50, at least 55, at least 60, at least 65, at least 7 0, at least 75, at least 80, at least 85, at least 90, at least 9 1, at least 92, at least 93, at least 94, at least 95, at least 9 6. Having at least 97%, at least 98%, or at least 99% sequence identity It includes an amino acid sequence or the sequence of SEQ ID NO: 10. In one embodiment, scFv is SEQ ID NO: 11 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 Having 96, at least 97, at least 98, or at least 99% sequence identity It includes an amino acid sequence or the sequence of SEQ ID NO. 11. In one embodiment, scFv is the sequence number No. 12 and at least 50, at least 55, at least 60, at least 65, and at least 70, at least 75, at least 80, at least 85, at least 90, and at least 91, at least 92, at least 93, at least 94, at least 95, and at least It also has 96%, at least 97%, at least 98%, or at least 99% sequence identity. It includes the amino acid sequence or the sequence of SEQ ID NO: 12. In one embodiment, scFv is the sequence Number 13 and at least 50, at least 55, at least 60, at least 65, less 70, at least 75, at least 80, at least 85, at least 90, less 91, at least 92, at least 93, at least 94, at least 95, less All have a sequence identity of 96%, at least 97%, at least 98%, or at least 99%. It contains an amino acid sequence, or the sequence of SEQ ID NO: 13. In one embodiment, scFv is distributed Column number 14 and at least 50, at least 55, at least 60, at least 65, less At least 70, at least 75, at least 80, at least 85, at least 90, less At least 91, at least 92, at least 93, at least 94, at least 95, less At least 96%, at least 97%, at least 98%, or at least 99% sequence identity It includes an amino acid sequence having, or the sequence of SEQ ID NO: 14. In one embodiment, scFv is Sequence ID 15 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 identical. It includes an amino acid sequence having a property, or the sequence of SEQ ID NO: 15. In one embodiment, scFv is , Sequence ID 16 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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% of the sequence It contains a monogeneous amino acid sequence or the sequence of SEQ ID NO: 16. In one embodiment, scFv This is sequence number 17 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, at least 80, at least 85, at least 90 , at least 91, at least 92, at least 93, at least 94, at least 95 , an array of at least 96, at least 97, at least 98, or at least 99% It contains an identical amino acid sequence, or the sequence of SEQ ID NO: 17. In one embodiment, scF v is sequence number 18 and at least 50, at least 55, at least 60, at least 6 5, at least 70, at least 75, at least 80, at least 85, at least 9 0, at least 91, at least 92, at least 93, at least 94, at least 9 5. At least 96, at least 97, at least 98, or at least 99% of the distribution It includes an amino acid sequence having column identity, or the sequence of SEQ ID NO: 18. In one embodiment, sc Fv is at least number 19 and at least 50, at least 55, at least 60, at 65, at least 70, at least 75, 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% It includes an amino acid sequence having sequence identity, or the sequence of SEQ ID NO: 19. In one embodiment, s cFv is at least sequence number 20 and at least 50, at least 55, at least 60, and 65, at least 70, at least 75, at least 80, at least 85, and at least 90, at least 91, at least 92, at least 93, at least 94, and at least 95, at least 96, at least 97, at least 98, or at least 99% It includes an amino acid sequence having sequence identity, or the sequence of SEQ ID NO: 20. In one embodiment, scFv is sequence number 21 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, less 90, at least 91, at least 92, at least 93, at least 94, less 95, at least 96, at least 97, at least 98, or at least 99 It includes an amino acid sequence having % sequence identity, or the sequence of SEQ ID NO: 21. In one embodiment, scFv is sequence number 22 and at least 50, at least 55, at least 60, less At least 65, at least 70, at least 75, at least 80, at least 85, less At least 90, at least 91, at least 92, at least 93, at least 94, less at least 95, at least 96, at least 97, at least 98, or at least 9 The amino acid sequence having 9% sequence identity, or the sequence of SEQ ID NO: 22, is included in one embodiment. scFv is sequence number 23 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 An amino acid sequence having 99% sequence identity, or the sequence of SEQ ID NO: 23, is included. One embodiment So, scFv is sequence number 169 and at least 50, at least 55, at least 60 , at least 65, at least 70, at least 75, 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 Both contain amino acid sequences with 99% sequence identity, or the sequence of SEQ ID NO: 169. In the application form, scFv is at least 50, at least 55, and at least number 170. Also 60, at least 65, at least 70, at least 75, at least 80, and at least 85, at least 90, at least 91, at least 92, at least 93, and at least 94, at least 95, at least 96, at least 97, at least 98, or An amino acid sequence having at least 99% sequence identity, or containing the sequence of SEQ ID NO: 170. In one embodiment, scFv is at least 50, at least 55, and less than 171. At least 60, at least 65, at least 70, at least 75, 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, Alternatively, an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 171. Includes. In one embodiment, scFv is sequence number 172 and at least 50, at least 5 5, at least 60, at least 65, at least 70, at least 75, at least 8 0, at least 85, at least 90, at least 91, at least 92, at least 9 3, at least 94, at least 95, at least 96, at least 97, at least 9 8, or an amino acid sequence having at least 99% sequence identity, or SEQ ID NO: 172 Includes an array of . In one embodiment, scFv includes at least array number 173 and 50, at least 55, at least 60, at least 65, at least 70, at least 75, less 80, at least 85, at least 90, at least 91, at least 92, less 93, at least 94, at least 95, at least 96, at least 97, at least Amino acid sequences or sequence numbers that have 98% or at least 99% sequence identity. It includes the sequence 173. In one embodiment, scFv includes sequence number 174 and at least 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, An amino acid sequence having at least 98, or at least 99%, sequence identity, or It includes the array with column number 174. In one embodiment, scFv includes array number 175 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 of amino acid sequences, or includes the sequence of sequence number 175. In one embodiment, scFv includes sequence number 176 and a small number of At least 50, at least 55, at least 60, at least 65, at least 70, less At least 75, at least 80, at least 85, at least 90, at least 91, less At least 92, at least 93, at least 94, at least 95, at least 96, less amino acids having at least 97, at least 98, or at least 99% sequence identity Includes an array, or the array of sequence number 176. In one embodiment, scFv is sequence number 177 and at least 50, at least 55, at least 60, at least 65, at least 70 , at least 75, 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 , having at least 97%, at least 98%, or at least 99% sequence identity It contains a mino acid sequence or the sequence of SEQ ID NO: 177. In one embodiment, scFv is SEQ ID NO: 1 78 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 Having 96, at least 97, at least 98, or at least 99% sequence identity It includes an amino acid sequence or the sequence of SEQ ID NO: 178. In one embodiment, scFv is a sequence Number 179 and at least 50, at least 55, at least 60, at least 65, fewer At least 70, at least 75, at least 80, at least 85, at least 90, less At least 91, at least 92, at least 93, at least 94, at least 95, less At least 96%, at least 97%, at least 98%, or at least 99% sequence identity The amino acid sequence having, or the sequence of SEQ ID NO: 179. In one embodiment, scFv is , Sequence ID 180 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, at least 80, at least 85, at least 90 , at least 91, at least 92, at least 93, at least 94, at least 95 , an array of at least 96, at least 97, at least 98, or at least 99% It contains an identical amino acid sequence, or the sequence of SEQ ID NO: 180. In one embodiment, sc Fv is at least 50, at least 55, at least 60, and at least 181. 65, at least 70, at least 75, at least 80, at least 85, and at least 90, at least 91, at least 92, at least 93, at least 94, and at least 95, at least 96, at least 97, at least 98, or at least 99% It includes an amino acid sequence having sequence identity, or the sequence of SEQ ID NO: 181. In one embodiment, scFv is sequence number 182 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 Contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 182. In this state, scFv is sequence number 183 and at least 50, at least 55, at least 6 0, at least 65, at least 70, at least 75, at least 80, at least 8 5, at least 90, at least 91, at least 92, at least 93, at least 9 4. At least 95, at least 96, at least 97, at least 98, or less It contains an amino acid sequence with at least 99% sequence identity, or the sequence of SEQ ID NO: 183. In this embodiment, scFv is at least 50, at least 55, and less than 184. 60, at least 65, at least 70, at least 75, at least 80, less 85, at least 90, at least 91, at least 92, at least 93, less 94, at least 95, at least 96, at least 97, at least 98, or It contains an amino acid sequence with at least 99% sequence identity, or the sequence of SEQ ID NO: 184. In one embodiment, scFv is sequence number 340 and at least 50, at least 55, At least 60, at least 65, at least 70, at least 75, 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, Alternatively, an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 340 Includes columns. In one embodiment, scFv includes sequence numbers 341 and at least 50, at 55, at least 60, at least 65, at least 70, at least 75, 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%, of amino acid sequences with sequence identity, or SEQ ID NO: 34 Includes an array of 1. In one embodiment, scFv includes at least arrays 342 and 50, and at least 50. At least 55, at least 60, at least 65, at least 70, at least 75, less At least 80, at least 85, at least 90, at least 91, at least 92, less At least 93, at least 94, at least 95, at least 96, at least 97, at least An amino acid sequence having at least 98, or at least 99%, sequence number Includes sequence number 342. In one embodiment, scFv includes sequence number 343 and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 , an amino acid sequence having at least 98, or at least 99%, sequence identity, Contains the sequence with sequence number 343.

[0279] In one embodiment, the extracellular antigen-binding domain includes a signal polypeptide. Polypeptides can be positioned at the N-terminus of the extracellular antigen-binding domain that binds to hK2. The signal polypeptide is extracellular during cell processing and localization of CAR to the cell membrane. The antigen-binding domain can be selectively cleaved. Various signaling molecules known to those skilled in the art Any of the thios may be used as the signal polypeptide. Non-limiting examples of peptides that may result include FcεR and human immunoglobulin (IgG) The variable region of the chain (HC), CD8α, or various other proteins secreted by T cells One of the following is a possible explanation. In various embodiments, the signal polypeptide acts as a secretory pathway for T cells. It is road and compatible. In some embodiments, the signal polypeptide is SEQ ID NO: 24 It contains an amino acid sequence. In one embodiment, the signal polypeptide is less than SEQ ID NO: 24. 50, at least 55, at least 60, at least 65, at least 70, less 75, at least 80, at least 85, at least 90, at least 91, less 92, at least 93, at least 94, at least 95, at least 96, less Both amino acid sequences have 97%, at least 98%, or at least 99% sequence identity. include.

[0280] In one embodiment, the disclosure relates to an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain. We provide a CAR containing a signaling domain, the extracellular antigen-binding domain of which is SEQ ID NO: 29 Select from the group consisting of ~44, 78~81, 221~236, and 273~308. Contains mino acid sequences. Sequence IDs 29-44, 78-81, 221-236, and 273-3 CA having an extracellular antigen-binding domain containing an amino acid sequence selected from the group consisting of 08 Another characteristic of R is that its extracellular antigen-binding domain binds to the hK2 antigen.

[0281] In one embodiment, the intracellular signaling domain is a TNF receptor superfamily The components of CD137 (CD9) and CD3 zeta chain (CD3z) of the T cell surface glycoprotein. , surface antigen classification (CD27) components, surface antigen classification superfamily members (e.g., Components such as CD28 or inducible T cell costimulator (ICOS), and combinations thereof. It contains a polypeptide component selected from the group consisting of the following:

[0282] In one embodiment, component CD137 includes the amino acid sequence of SEQ ID NO: 27. So, the CD137 component is at least SEQ ID NO: 27 and at least 50, at least 55, at 60, at least 65, at least 70, at least 75, 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 less Both contain amino acid sequences with 99% sequence identity.

[0283] In one embodiment, the CD3z component includes the amino acid sequence of SEQ ID NO: 28. The CD3z component is at least 50, at least 55, and at least 60 of sequence numbers 28 and 50. , at least 65, at least 70, at least 75, 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 It contains an amino acid sequence with 99% sequence identity.

[0284] In one embodiment, the intracellular signaling domain includes the amino acid sequence of SEQ ID NO: 45 In one embodiment, the intracellular signaling domain is sequence number 45 and at least 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, It contains an amino acid sequence with at least 98, or at least 99%, sequence identity.

[0285] In one embodiment, the transmembrane domain is a CD8a transmembrane region (CD8A-TM) polypeptide Contains thiosulfate. In some embodiments, the CD8a-TM polypeptide is the same as in SEQ ID NO: 26 It contains an amino acid sequence. In some embodiments, the CD8a-TM polypeptide is sequence number No. 26 and at least 50, at least 55, at least 60, at least 65, and at least 70, at least 75, at least 80, at least 85, at least 90, and at least 91, at least 92, at least 93, at least 94, at least 95, and at least It also has 96%, at least 97%, at least 98%, or at least 99% sequence identity. Contains amino acid sequence.

[0286] In one embodiment, the transmembrane domain is the α, β, or ζ chain of the T cell receptor, CD28, C D3 Epsilon, CD45, CD4, CD5, CD8, CD8α, CD9, CD16, C D22, CD33, CD37, CD40, CD64, CD80, CD86, CD134, It includes at least the transmembrane regions of CD137 and CD154. In another embodiment, transmembrane domain In is ζ, η, or FcεR1γ and -β, MB1(Igα), B29, or CD3-γ It includes at least a transmembrane domain of ζ or η. In another embodiment, the transmembrane domain The compound is synthetic, for example, a triplet of leucine and valine, and phenylalanine. It contains mainly hydrophobic residues such as tryptophan.

[0287] In one embodiment, the CAR has a transmembrane domain linked to an extracellular antigen-binding domain. It further includes a hinge region. In some embodiments, the hinge region is the CD8a hinge region. In some embodiments, the CD8a-hinge region includes the amino acid sequence of SEQ ID NO: 25. In some embodiments, the CD8a-hinge region is at least 50 of sequence number 25. , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 It contains an amino acid sequence having at least 98, or at least 99%, sequence identity. In some embodiments, the hinge region is array EPKSCDKTHTCPPCP(array array). 269) includes or at least EPKSCDKTHTCPPCP(sequence ID 269) Also 50, at least 55, at least 60, at least 65, at least 70, and 75, at least 80, at least 85, at least 90, at least 91, and at least 92, at least 93, at least 94, at least 95, at least 96, and at least It also includes an amino acid sequence having 97, at least 98, or at least 99% sequence identity. In some embodiments, the hinge region is array ERKCCVECPPCP(array sequence number). 270) includes or contains ERKCCVECPPCP (Sequence ID 270) and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 It contains an amino acid sequence having at least 98, or at least 99%, sequence identity. In some embodiments, the hinge region is the array ELKTPLGDTTHTCPRCP(EP Includes KSCDTPPPCPRCP)3 (Sequence ID 271) or ELKTPLGDT THTCPRCP(EPKSCDTPPPCPRCP)3 (Sequence ID 271) and at least Also 50, at least 55, at least 60, at least 65, at least 70, and 75, at least 80, at least 85, at least 90, at least 91, and at least 92, at least 93, at least 94, at least 95, at least 96, and at least It also includes an amino acid sequence having 97, at least 98, or at least 99% sequence identity. Hmm. In some embodiments, the hinge region is array ESKYGPPCPSCP(array array). 272) includes or ESKYGPPCPSCP (Sequence ID 272) and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 , comprising an amino acid sequence having at least 98, or at least 99%, sequence identity.

[0288] g) In one embodiment, the CAR comprises an extracellular antigen-binding domain, a hinge region, and a transmembrane domain. It includes a , and an intracellular signaling domain. In such one embodiment, the hinge region is The CD8α hinge region is the CD8α-TM domain, and the transmembrane domain is the cell The internal signal transduction domains are the CD3-zeta domain and the 4-1BB / CD137 domain. It includes n.

[0289] h) In such one embodiment, the hinge region includes the amino acid sequence of SEQ ID NO: 25. The D8α hinge region and the transmembrane domain contain the amino acid sequence of SEQ ID NO: 26. The α-TM domain is the intracellular signaling domain, and the CD3-zeta domain and It contains the 4-1BB / CD137 domain, which includes the amino acid sequence of SEQ ID NO: 45.

[0290] i) In another embodiment, the hinge region is CD8α containing the amino acid sequence of SEQ ID NO: 25 The transmembrane domain is the CD8α-TM region, which contains the amino acid sequence of SEQ ID NO: 26. The domain, and the intracellular signaling domain, contains 4 amino acids, including the amino acid sequence of SEQ ID NO: 27. CD3-zetad containing the -1BB / CD137 domain and the amino acid sequence of SEQ ID NO: 28 Includes the main course.

[0291] In one embodiment, the extracellular antigen-binding domain is sequence numbers 29-44, 78-81, 22 The amino acid sequence includes an amino acid sequence selected from the groups consisting of 1-236 and 273-308. Morphologically, the extracellular antigen-binding domain is at least in SEQ ID NOs. 29 and 50, and at least in SEQ ID NOs. 55 , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 29 Includes columns. In one embodiment, the extracellular antigen-binding domain is sequence number 30 and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 , an amino acid sequence having at least 98, or at least 99%, sequence identity, It includes the sequence of SEQ ID NO: 30. In one embodiment, the extracellular antigen-binding domain is SEQ ID NO: 31 and at least 50, at least 55, at least 60, at least 65, at least 70 , at least 75, 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 , having at least 97%, at least 98%, or at least 99% sequence identity It contains a mino acid sequence or the sequence of SEQ ID NO: 31. In one embodiment, it has an extracellular antigen-binding domain. This is sequence number 32 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, at least 80, at least 85, at least 90 , at least 91, at least 92, at least 93, at least 94, at least 95 , an array of at least 96, at least 97, at least 98, or at least 99% It contains an identical amino acid sequence, or the sequence of SEQ ID NO: 32. In one embodiment, it is extracellular. The antigen-binding domains are at least SEQ ID NO: 33, at least 50, at least 55, and at least 60. , at least 65, at least 70, at least 75, 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 Both contain amino acid sequences with 99% sequence identity, or the sequence of SEQ ID NO: 33. Morphologically, the extracellular antigen-binding domain is at least in SEQ ID NOs. 34 and 50, and at least in SEQ ID NOs. 55 , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 34 Includes columns. In one embodiment, the extracellular antigen-binding domain is sequence number 35 and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 , an amino acid sequence having at least 98, or at least 99%, sequence identity, It includes the sequence of SEQ ID NO: 35. In one embodiment, the extracellular antigen-binding domain is SEQ ID NO: 36 and at least 50, at least 55, at least 60, at least 65, at least 70 , at least 75, 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 , having at least 97%, at least 98%, or at least 99% sequence identity It contains a mino acid sequence or the sequence of SEQ ID NO: 36. In one embodiment, it contains an extracellular antigen-binding domain. This is sequence number 37 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, at least 80, at least 85, at least 90 , at least 91, at least 92, at least 93, at least 94, at least 95 , an array of at least 96, at least 97, at least 98, or at least 99% It contains an identical amino acid sequence, or the sequence of SEQ ID NO: 37. In one embodiment, it is extracellular. The antigen-binding domain is at least sequence numbers 38, 50, 55, and 60. , at least 65, at least 70, at least 75, 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 Both contain amino acid sequences with 99% sequence identity, or the sequence of SEQ ID NO: 38. Morphologically, the extracellular antigen-binding domain is at least in SEQ ID NOs. 39 and 50, and at least in SEQ ID NOs. 55 , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 39 Includes columns. In one embodiment, the extracellular antigen-binding domain is sequence number 40 and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 , an amino acid sequence having at least 98, or at least 99%, sequence identity, It includes the sequence of SEQ ID NO: 40. In one embodiment, the extracellular antigen-binding domain is SEQ ID NO: 41 and at least 50, at least 55, at least 60, at least 65, at least 70 , at least 75, 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 , having at least 97%, at least 98%, or at least 99% sequence identity It contains a mino acid sequence or the sequence of SEQ ID NO: 41. In one embodiment, it has an extracellular antigen-binding domain. This is sequence number 42 and at least 50, at least 55, at least 60, at least 65 , at least 70, at least 75, at least 80, at least 85, at least 90 , at least 91, at least 92, at least 93, at least 94, at least 95 , an array of at least 96, at least 97, at least 98, or at least 99% It contains an identical amino acid sequence, or the sequence of SEQ ID NO: 42. In one embodiment, it is extracellular. The antigen-binding domains are at least SEQ ID NOs. 43, 50, 55, and 60. , at least 65, at least 70, at least 75, 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 Both contain amino acid sequences with 99% sequence identity, or the sequence of SEQ ID NO: 43. Morphologically, the extracellular antigen-binding domain is at least in SEQ ID NOs. 44 and 50, and at least in SEQ ID NOs. 55 , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 44 Includes a column. In one embodiment, the extracellular antigen-binding domain is sequence number 78 and at least 50 , at least 55, at least 60, at least 65, at least 70, at least 75 , 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 , an amino acid sequence having at least 98, or at least 99%, sequence identity, It includes the sequence of SEQ ID NO: 78. In one embodiment, the extracellular antigen-binding domain is SEQ ID NO: 79 and at least 50, at least 55, at least 60, at least 65, at least 70 , at least 75, 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 , having at least 97%, at least 98%, or at least 99% sequence identity It contains a mino acid sequence or the sequence of Sequence ID No. 79. In one embodiment, it has an extracellular antigen-binding domain. This includes at least sequence numbers 80 and at least 50, at least 55, at least 60, and at least 65. , at least 70, at least 75, at least 80, at least 85, at least 90 , at least 91, at least 92, at least 93, at least 94, at least 95 , an array of at least 96, at least 97, at least 98, or at least 99% It contains an identical amino acid sequence, or the sequence of SEQ ID NO: 80. In one embodiment, it is extracellular. The antigen-binding domain is sequence number 81 and at least 50, at least 55, at least 60 , at least 65, at least 70, at least 75, 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 Both contain amino acid sequences with 99% sequence identity, or the sequence of SEQ ID NO: 81. Morphologically, the extracellular antigen-binding domain is at least 50 of sequence numbers 221 and 50. 5, at least 60, at least 65, at least 70, at least 75, at least 8 0, at least 85, at least 90, at least 91, at least 92, at least 9 3, at least 94, at least 95, at least 96, at least 97, at least 9 8, or an amino acid sequence having at least 99% sequence identity, or SEQ ID NO: 221 The sequence includes. In one embodiment, the extracellular antigen-binding domain is at least SEQ ID NO: 222. Also 50, at least 55, at least 60, at least 65, at least 70, and 75, at least 80, at least 85, at least 90, at least 91, and at least 92, at least 93, at least 94, at least 95, at least 96, and at least Amino acid sequences having 97%, at least 98%, or at least 99% sequence identity , or includes the sequence of Sequence ID No. 222. In one embodiment, the extracellular antigen-binding domain is the sequence Number 223 and at least 50, at least 55, at least 60, at least 65, less At least 70, at least 75, at least 80, at least 85, at least 90, less At least 91, at least 92, at least 93, at least 94, at least 95, less At least 96%, at least 97%, at least 98%, or at least 99% sequence identity The amino acid sequence having, or the sequence of SEQ ID NO: 223. In one embodiment, an extracellular antigen The binding domain is sequence number 224 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 It also contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 224. Morphologically, the extracellular antigen-binding domain is at least 50 of SEQ ID NO: 225, at least 5 5 , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 225 Includes a sequence. In one embodiment, the extracellular antigen-binding domain is sequence number 226 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 of amino acid sequences, or includes the sequence of sequence number 226. In one embodiment, the extracellular antigen-binding domain is sequence number No. 227 and at least 50, at least 55, at least 60, at least 65, less 70, at least 75, at least 80, at least 85, at least 90, less 91, at least 92, at least 93, at least 94, at least 95, less All have a sequence identity of 96%, at least 97%, at least 98%, or at least 99%. It contains an amino acid sequence, or the sequence of SEQ ID NO: 227. In one embodiment, extracellular antigen cluster The combined domain is sequence number 228 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 Contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 228. In this state, the extracellular antigen-binding domain is at least 50 and at least 55 of sequence number 229. , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or sequence number 229 Includes a sequence. In one embodiment, the extracellular antigen-binding domain is sequence number 230 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 of amino acid sequences, or includes the sequence of sequence number 230. In one embodiment, the extracellular antigen-binding domain is sequence number No. 231 and at least 50, at least 55, at least 60, at least 65, less 70, at least 75, at least 80, at least 85, at least 90, less 91, at least 92, at least 93, at least 94, at least 95, less All have a sequence identity of 96%, at least 97%, at least 98%, or at least 99%. It contains an amino acid sequence, or the sequence of SEQ ID NO: 231. In one embodiment, it contains an extracellular antigen The combined domain is sequence number 232 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 Contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 232. One implementation form In this state, the extracellular antigen-binding domain is at least 50 and at least 55 of sequence number 233. , at least 60, at least 65, at least 70, at least 75, 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 an amino acid sequence having at least 99% sequence identity, or sequence number 233 Includes a sequence. In one embodiment, the extracellular antigen-binding domain is sequence number 234 and at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, 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 of amino acid sequences, or includes the sequence of sequence number 234. In one embodiment, the extracellular antigen-binding domain is sequence number No. 235 and at least 50, at least 55, at least 60, at least 65, less 70, at least 75, at least 80, at least 85, at least 90, less 91, at least 92, at least 93, at least 94, at least 95, less All have a sequence identity of 96%, at least 97%, at least 98%, or at least 99%. It contains an amino acid sequence, or the sequence of SEQ ID NO: 235. In one embodiment, it contains an extracellular antigen The combined domain is sequence number 236 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 It contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 236.

[0292] CAR constructs and immune-responsive cells expressing CARs The term "coding" refers to a defined sequence of nucleotides (e.g., rR A biological organism having any of the defined sequences of amino acids (NA, tRNA, and mRNA) or amino acids. It serves as a template for the synthesis of other polymers and macromolecules in scientific processes. To identify nucleotides in polynucleotides such as genes, cDNA, or mRNA. This refers to the inherent characteristics of the sequence of cells, as well as the biological characteristics arising therefrom. Therefore, cells or In other biological systems, the transcription and translation of mRNA corresponding to genes produce proteins. In this case, the gene, cDNA, or RNA codes for a protein. A coding strand whose columns are identical to the mRNA sequence, and a transcription template for a gene or cDNA. Any non-coding strand used in this way is a protein or other product of the gene or cDNA. It can be referred to as coding things.

[0293] Unless otherwise stated, "nucleotide sequences encoding amino acid sequences" are mutually degenerate. All nucleotide sequences that are of the same type, and all nucleotide sequences that encode the same amino acid sequence. To include. The phrase "nucleotide sequence encoding a protein or RNA" also includes several. In that case, the nucleotide sequence encoding the protein may contain introns. It can encompass introns to a certain extent.

[0294] The term "expression vector" refers to a vector that is operablely linked to the nucleotide sequence to be expressed. This refers to a vector containing recombinant polynucleotides that include expression regulatory sequences. It contains sufficient cis-acting elements for expression, and other elements for expression are present in the host cell. Therefore, it can be supplied in an in vitro expression system. As an expression vector, recombinant Cosmids and plasmids incorporating polynucleotides (for example, naked or liposomes) (containers), and viruses (e.g., lentiviruses, retroviruses, adenoids) All vectors known in the art, including viruses and adeno-associated viruses. These are some examples.

[0295] In one embodiment, the present disclosure provides cells that express nucleic acid molecules encoding CARs. In one embodiment, the CARs of this disclosure are sequence numbers 46-61, 82-85, and 20. It contains an amino acid sequence selected from the group consisting of 5 to 220. In some embodiments, The disclosed CARs are sequence numbers 46 and at least 50, at least 55, at least 60, and At least 65, at least 70, at least 75, 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 It contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 46. In embodiments, the CAR of this disclosure includes at least SEQ ID NOs. 47 and 50, at least 55, and a small number. At least 60, at least 65, at least 70, at least 75, 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, Alternatively, an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 47 Includes. In some embodiments, the CARs of this disclosure include at least SEQ ID NOs. 48 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, An amino acid sequence having at least 98, or at least 99%, sequence identity, or sequence Includes sequence number 48. In some embodiments, the CAR of this disclosure includes sequence number 49 and a small number of others. At least 50, at least 55, at least 60, at least 65, at least 70, At least 75, 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, amino acids having at least 97%, at least 98%, or at least 99% sequence identity. The acid sequence or the sequence of SEQ ID NO: 49 is included. In some embodiments, the CAR of the present disclosure is Attenuation Cavity 50 and at least 50, at least 55, at least 60, at least 65, few At least 70, at least 75, 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 identical. The amino acid sequence having the property, or the sequence of SEQ ID NO: 50. In some embodiments, this The disclosed CARs are sequence numbers 51 and at least 50, at least 55, at least 60, At least 65, at least 70, at least 75, 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 It contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 51. In embodiments, the CAR of the present disclosure includes at least 50, at least 55, and at least 52. At least 60, at least 65, at least 70, at least 75, 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, Alternatively, an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 52 Includes. In some embodiments, the CARs of this disclosure include at least SEQ ID NO: 53 and 50, At least 55, at least 60, at least 65, at least 70, at least 75, 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, An amino acid sequence having at least 98, or at least 99%, sequence identity, or sequence Includes sequence number 53. In some embodiments, the CAR of the present disclosure includes sequence number 54 and a small number of others. At least 50, at least 55, at least 60, at least 65, at least 70, At least 75, 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, amino acids having at least 97%, at least 98%, or at least 99% sequence identity. The acid sequence or the sequence of SEQ ID NO: 54 is included. In some embodiments, the CAR of the present disclosure is Attenuation Cavity 55 and at least 50, at least 55, at least 60, at least 65, At least 70, at least 75, 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 identical. The amino acid sequence having the property, or the sequence of SEQ ID NO: 55. In some embodiments, this The disclosed CARs are sequence numbers 56 and at least 50, at least 55, at least 60, and At least 65, at least 70, at least 75, 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 It contains an amino acid sequence with 99% sequence identity, or the sequence of SEQ ID NO: 56. In embodiments, the CAR of the present disclosure includes at least 50, at least 55, and at least 57. At least 60, at least 65, at least 70, at least 75, 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, Alternatively, an amino acid sequence having at least 99% sequence identity, or the sequence of SEQ ID NO: 57 Includes. In some embodiments, the CARs of this disclosure include at lea...

Claims

1. It is a chimeric antigen receptor (CAR), (a) an extracellular domain that specifically binds to the human kallikrein-2 (hK2) antigen, (b) Transmembrane domain and (c) Intracellular signaling domains optionally containing at least one co-stimulatory domain and, CAR, including.

2. A CAR according to claim 1, (d) CD8a - Hinge region It further includes, The transmembrane domain comprises a CD8a transmembrane region (CD8a-TM) polypeptide; The intracellular signaling domain is a TNF receptor superfamily member 9(C) D137) Co-stimulatory domain containing component and T cell surface glycoprotein CD3 zeta chain (CD A CAR containing a primary signaling domain that includes the 3z component.

3. A CAR according to claim 2, The CD8a-hinge region corresponds to sequence numbers 25, 269, 270, 271, or 27 Containing an amino acid sequence that is at least 90% identical to 2, The transmembrane domain contains an amino acid sequence that is at least 90% identical to that of SEQ ID NO:

26. mu, and / or The intracellular signaling domain is at least 90% identical to that of SEQ ID NO:

27. A co-stimulatory domain having an acid sequence, and an amino acid sequence that is at least 90% identical to SEQ ID NO 28. A CAR containing a primary signaling domain with an acid sequence.

4. Including an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain, A chimeric antigen receptor (CAR), wherein the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 4, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, Heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 315, and the amino acid sequence of SEQ ID NO: 311 Heavy chain CDR3 having rows; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 309, SEQ ID NO: A heavy chain CDR2 having a 310-amino acid sequence, and having the amino acid sequence of SEQ ID NO:

311. heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 72, SEQ ID NO: 7 Heavy chain CDR2 having the amino acid sequence of 3, and heavy chain having the amino acid sequence of SEQ ID NO: 66 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 86, SEQ ID NO: 8 Heavy chain CDR2 having the amino acid sequence of 7, and heavy chain having the amino acid sequence of SEQ ID NO: 88 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 92, SEQ ID NO: 9 Heavy chain CDR2 having the amino acid sequence of 3, and heavy chain having the amino acid sequence of SEQ ID NO: 94 CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: A heavy chain CDR2 having the amino acid sequence of 103, and having the amino acid sequence of SEQ ID NO: 104 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: A heavy chain CDR2 having a sequence of 106 amino acids, and having the amino acid sequence of SEQ ID NO: 104 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: A heavy chain CDR2 having a sequence of 108 amino acids, and having the amino acid sequence of SEQ ID NO: 109 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 110, SEQ ID NO: A heavy chain CDR2 having an amino acid sequence of 111, and having the amino acid sequence of SEQ ID NO: 109 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: A heavy chain CDR2 having a sequence of 112 amino acids, and having the amino acid sequence of SEQ ID NO: 113 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 105, SEQ ID NO: A heavy chain CDR2 having a sequence of 114 amino acids, and having the amino acid sequence of SEQ ID NO: 113 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: A heavy chain CDR2 having a sequence of 116 amino acids, and having the amino acid sequence of SEQ ID NO: 117 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 118, SEQ ID NO: A heavy chain CDR2 having a sequence of 119 amino acids, and having the amino acid sequence of SEQ ID NO: 117 heavy chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: A heavy chain CDR2 having a 121-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 122 heavy chain CDR3; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 123, SEQ ID NO: A heavy chain CDR2 having a 124-amino acid sequence, and having the amino acid sequence of SEQ ID NO: 122 heavy chain CDR3, Includes, The extracellular antigen-binding domain is a CAR that binds to the hK2 antigen.

5. Including an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain, A chimeric antigen receptor (CAR), wherein the extracellular antigen-binding domain is Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 6 Light chain CDR2 having the amino acid sequence of 9, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, SEQ ID NO: 7 Light chain CDR2 having the amino acid sequence 0, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence 0, and light chain having the amino acid sequence of SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 95, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence of 6, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 312, SEQ ID NO: A light chain CDR2 having a sequence of 313 amino acids, and having the amino acid sequence of SEQ ID NO: 91 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence 0, and light chain having the amino acid sequence of SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 1 Light chain CDR2 having a 26-amino acid sequence, and light chain having the amino acid sequence of SEQ ID NO: 71 chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 128, SEQ ID NO: A light chain CDR2 having a sequence of 129 amino acids, and having the amino acid sequence of SEQ ID NO:

130. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 125, SEQ ID NO: A light chain CDR2 having the amino acid sequence of 131, and having the amino acid sequence of SEQ ID NO: 132 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 133, SEQ ID NO: A light chain CDR2 having a sequence of 134 amino acids, and having the amino acid sequence of SEQ ID NO: 135 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 136, SEQ ID NO: A light chain CDR2 having a sequence of 137 amino acids, and having the amino acid sequence of SEQ ID NO: 138 Light chain CDR3; or Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 97, SEQ ID NO: 1 A light chain CDR2 having a sequence of 27 amino acids, and having the amino acid sequence of SEQ ID NO: 139 Light chain CDR3, Includes, The extracellular antigen-binding domain of the CAR binds to the hK2 antigen.

6. The CAR according to claim 4, wherein the extracellular antigen-binding domain is Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 6 Light chain CDR2 having the amino acid sequence of 9, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, SEQ ID NO: 7 Light chain CDR2 having the amino acid sequence 0, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence 0, and light chain having the amino acid sequence of SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 95, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence of 6, and light chain having the amino acid sequence of SEQ ID NO: 71 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 89, SEQ ID NO: 9 Light chain CDR2 having the amino acid sequence 0, and light chain having the amino acid sequence of SEQ ID NO: 91 CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, SEQ ID NO: 1 Light chain CDR2 having a 26-amino acid sequence, and light chain having the amino acid sequence of SEQ ID NO: 71 chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 128, SEQ ID NO: A light chain CDR2 having a sequence of 129 amino acids, and having the amino acid sequence of SEQ ID NO:

130. Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 125, SEQ ID NO: A light chain CDR2 having the amino acid sequence of 131, and having the amino acid sequence of SEQ ID NO: 132 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 133, SEQ ID NO: A light chain CDR2 having a sequence of 134 amino acids, and having the amino acid sequence of SEQ ID NO: 135 Light chain CDR3; Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 136, SEQ ID NO: A light chain CDR2 having a sequence of 137 amino acids, and having the amino acid sequence of SEQ ID NO: 138 Light chain CDR3; or Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 97, SEQ ID NO: 1 A light chain CDR2 having a sequence of 27 amino acids, and having the amino acid sequence of SEQ ID NO: 139 Light chain CDR3, CAR, which further includes...

7. The CAR according to claim 6, wherein the extracellular antigen-binding domain is Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 4, and heavy chain CD having the amino acid sequence of SEQ ID NO:

66. R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain CD having the amino acid sequence of SEQ ID NO:

66. R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 68, Light chain CDR2 having the amino acid sequence of sequence number 70, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain CD having the amino acid sequence of SEQ ID NO:

66. R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain CD having the amino acid sequence of SEQ ID NO:

66. R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 69, and having the amino acid sequence of sequence number 71 Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 63, SEQ ID NO: 6 Heavy chain CDR2 having the amino acid sequence of 5, and heavy chain CD having the amino acid sequence of SEQ ID NO:

66. R3 and light chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 67, Light chain CDR2 having the amino acid sequence of sequence number 126, and having the amino acid sequence of sequence number 71. Light chain CDR3; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 314, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 315, heavy chain having an amino acid sequence of SEQ ID NO: 311 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of chain CDR3 and SEQ ID NO: 312 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 313, and the amino acid sequence of SEQ ID NO: 91 Light chain CDR3 having a row; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having amino acid sequence 103, heavy chain having amino acid sequence 104 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of chain CDR3 and sequence number 128 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 129, and the amino acids of SEQ ID NO: 130 Light chain CDR3 having an arrangement; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 107, SEQ ID NO: Heavy chain CDR2 having a sequence of 108 amino acids, heavy chain having the amino acid sequence of SEQ ID NO: 109 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of chain CDR3 and sequence number 125 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 131, and the amino acids of SEQ ID NO: 132 Light chain CDR3 having an arrangement; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 102, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 112, heavy chain having an amino acid sequence of SEQ ID NO: 113 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of chain CDR3 and SEQ ID NO: 133 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 134, and the amino acids of SEQ ID NO: 135 Light chain CDR3 having an arrangement; Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 115, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 116, heavy chain having an amino acid sequence of SEQ ID NO: 117 Light chain complementarity determination region 1 (CDR3) having the amino acid sequence of chain CDR3 and SEQ ID NO: 136 1) Light chain CDR2 having the amino acid sequence of SEQ ID NO: 137, and the amino acids of SEQ ID NO: 138 Light chain CDR3 having an arrangement; or Heavy chain complementarity determination region 1 (CDR1) having the amino acid sequence of SEQ ID NO: 120, SEQ ID NO: Heavy chain CDR2 having an amino acid sequence of 121, heavy chain having an amino acid sequence of SEQ ID NO: 122 Light chain complementarity determination region 1 (CDR1) having the amino acid sequence of chain CDR3 and SEQ ID NO: 97 ), light chain CDR2 having the amino acid sequence of SEQ ID NO: 127, amino acid sequence of SEQ ID NO: 138 A CAR comprising a light chain CDR3 having a row.

8. The extracellular antigen-binding domain has the amino acid sequence of SEQ ID NO: 86, which determines heavy chain complementarity. Region 1 (CDR1), heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 87, SEQ ID NO: 88 A heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 89, and a light chain phase having the amino acid sequence of SEQ ID NO:

89. Complementarity-determining region 1 (CDR1), light chain CDR2 having the amino acid sequence of SEQ ID NO: 90, sequence The CAR according to claim 6, comprising a light chain CDR3 having amino acid sequence number 91.

9. Including an extracellular antigen-binding domain, a transmembrane domain, and an intracellular signaling domain, A chimeric antigen receptor (CAR), wherein the extracellular antigen-binding domain is Sequence IDs 1, 2, 3, 74, 75, 147, 148, 149, 150, 151, 152 , and a light chain variable domain (VL) comprising an amino acid sequence selected from 318, and / or teeth Sequence numbers 4, 5, 6, 76, 77, 140, 141, 142, 143, 144, 145 A heavy chain variable domain (VH) comprising an amino acid sequence selected from 146 and 317, Includes, The extracellular antigen-binding domain binds to the human kallikrein-2 (hK2) antigen. AR.

10. The CAR according to claim 9, wherein the extracellular antigen-binding domain is Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 4 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 6 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 2 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 3 and heavy chain variable region containing the amino acid sequence of SEQ ID NO: 5 Chain variable region; The light chain variable region includes the amino acid sequence of SEQ ID NO: 74 and the amino acid sequence of SEQ ID NO:

76. Heavy chain variable region; The light chain variable region includes the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO:

77. Heavy chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 147 and amino acid sequence of SEQ ID NO: 140 Heavy chain variable region including; The light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 141 Includes heavy chain variable region; The light chain variable region containing the amino acid sequence of SEQ ID NO: 75 and the amino acid sequence of SEQ ID NO: 140 Includes heavy chain variable region; Light chain variable region containing the amino acid sequence of SEQ ID NO: 148 and amino acid sequence of SEQ ID NO: 142 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 149 and amino acid sequence of SEQ ID NO: 143 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 150 and amino acid sequence of SEQ ID NO: 144 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 151 and amino acid sequence of SEQ ID NO: 145 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 152 and amino acid sequence of SEQ ID NO: 146 Heavy chain variable region including; Light chain variable region containing the amino acid sequence of SEQ ID NO: 318 and amino acid sequence of SEQ ID NO: 317 Heavy chain variable region including, CAR, including.

11. The extracellular antigen-binding domain comprises a single-stranded variable fragment (scFv), and the scFv is , comprising a light chain variable region (VL) and a heavy chain variable region (VH), any one of claims 1 to 10 The CAR as described.

12. The scFv is a phosphorus between the light chain variable region (VL) and the heavy chain variable region (VH). The CAR according to claim 11, comprising a car polypeptide.

13. The linker polypeptide comprises the amino acid sequence of SEQ ID NO: 7, according to claim 12. CAR.

14. The scFv is a group consisting of sequence numbers 8-23, 169-184, and 340-343. A CAR according to any one of claims 11 to 13, comprising an amino acid sequence selected from.

15. The extracellular antigen-binding domain comprises a signal polypeptide, according to any of claims 1 to 14. The CAR listed on one of the cards.

16. The signal polypeptide comprises the amino acid sequence of SEQ ID NO: 24, according to claim 15. The CAR.

17. The intracellular signaling domain is a TNF receptor superfamily member 9(C) D137) component, T cell surface glycoprotein CD3 zeta chain (CD3z) component, surface antigen component (CD27) components, surface antigen classification superfamily member components, and combinations thereof A polypeptide component selected from the group consisting of combinations, according to any one of claims 4 to 16 The CAR as described.

18. The CA according to claim 17, wherein the CD137 component includes the amino acid sequence of SEQ ID NO:

27. R.

19. The CD3z component comprises the amino acid sequence of SEQ ID NO: 28, according to claim 17 or 18. The CAR.

20. The intracellular signaling domain includes the amino acid sequence of SEQ ID NO: 45, or any other In the selection, the intracellular signaling domain contains the amino acid sequence of SEQ ID NO: 27 as a co-stimulatory element. A claim including a primary signaling domain containing the domain and the amino acid sequence of SEQ ID NO:

28. A CAR as described in any of items 17 to 19.

21. The transmembrane domain includes a CD8a transmembrane region (CD8a-TM) polypeptide. A CAR according to any one of claims 4 to 20.

22. Claim 21, wherein the CD8a-TM polypeptide comprises the amino acid sequence of SEQ ID NO: 26 The CAR listed.

23. The transmembrane domain further comprises a hinge region that connects the transmembrane domain to the extracellular antigen-binding domain, A CAR according to any one of claims 4 to 22.

24. The CAR according to claim 23, wherein the hinge region is the CD8a-hinge region.

25. The CD8a-hinge region is the same as sequence numbers 25, 269, 270, 271, or 272. The CAR according to claim 24, comprising an amino acid sequence.

26. The extracellular antigen-binding domain is sequence numbers 29-44, 78-81, 221-236, and an amino acid sequence selected from the group consisting of 273 to 308, as described in claims 1 to 25. The CAR listed in any of the above.

27. A net selected from the group consisting of sequence numbers 46-61, 82-85, and 205-220. A CAR according to any one of claims 1 to 26, comprising a no-acid sequence.

28. Isolated lymphocytes expressing the CAR described in any one of claims 1 to 27.

29. The isolated lymphocyte according to claim 28, which is a T lymphocyte.

30. The isolated lymphocyte according to claim 28, which is a natural killer (NK) cell.

31. An isolated nucleic acid molecule encoding the CAR described in any of claims 1 to 27.

32. A vector comprising the nucleic acid molecule described in claim 31.

33. A cell expressing the nucleic acid molecule described in claim 31.

34. A pharmaceutical composition comprising an effective amount of lymphocytes according to any one of claims 28 to 30.

35. An effective amount of lymphocytes according to any one of claims 28 to 30 and a pharmaceutically acceptable excipient A pharmaceutical composition containing the following.

36. A CAR according to any one of claims 1 to 27 for use in treatment, or claim The pharmaceutical composition described in 34 or 35.

37. A method for treating a subject with cancer, according to any one of claims 1 to 27 CAR, or the pharmaceutical composition according to claim 34 or 35.

38. A method for treating a subject with cancer, A therapeutically effective amount of lymphocytes according to any one of claims 28 to 30, which is required The substance is administered to the target, thereby inducing the lymphocytes to kill cancer cells in the target. A method that includes [this].

39. The method according to claim 38, wherein the cancer is prostate cancer or androgen receptor-expressing breast cancer. or a CAR or pharmaceutical composition for use in the method according to claim 37.

40. A method for targeting and killing cancer cells, The cancer cells are brought into contact with the lymphocytes described in any one of claims 28 to 30, thereby, A method comprising the lymphocytes inducing the killing of the cancer cells.

41. Claim 40, wherein the cancer cells are prostate cancer cells or androgen receptor-expressing breast cancer cells. Methods used.

42. A method for detecting the presence of cancer in a subject, The cell sample obtained from the subject is brought into contact with the CAR according to any one of claims 1 to 27. This causes the formation of a CAR-cell complex, The detection of the complex, where the detection of the complex indicates the presence of cancer in the subject. To indicate presence Methods that include...

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