Embryonic stem cell-derived tumor antigen epitope peptides and uses thereof

Abstract

This invention discloses tumor antigen epitope peptides derived from embryonic stem cells and their applications. The tumor antigen epitope peptides comprise one or more sequences selected from those shown in SEQ ID NO:1-10. This invention also provides antitumor pharmaceutical compositions comprising the aforementioned tumor antigen epitope peptides. This invention screens for highly expressed tumor antigens and their epitope peptides by comparing genes co-expressed by embryonic stem cells and tumor cells. Animal experiments have demonstrated that the extended peptides of these epitope peptides can stimulate specific cytotoxic T lymphocyte immune responses, stimulate the secretion of high levels of IFN-γ, and inhibit tumor growth.

Description

Technical Field

[0001] This invention relates to the field of biopharmaceutical technology, and in particular to tumor antigen epitope peptides derived from embryonic stem cells and their applications. Background Technology

[0002] Bladder cancer is a malignant tumor that occurs on the bladder mucosa. It is the most common malignant tumor of the urinary system and one of the ten most common cancers in the body. Bladder cancer generally occurs in the epithelial cells of the bladder, with urothelial carcinoma being the most common type, accounting for approximately 90%. Currently, the main treatments for bladder cancer are traditional methods (surgery and chemotherapy) and non-specific vaccine therapy such as BCG. Compared to traditional treatments, targeted therapy and immunotherapy are gaining popularity due to their high specificity and fewer side effects.

[0003] Tumor immunotherapy refers to a therapy that activates the body's immune system, relying on its own immune function to kill tumor cells and tissues. Unlike traditional surgery, chemotherapy, and radiotherapy, the direct target of tumor immunotherapy is not tumor tissue, but the body's own immune system. Tumor vaccines, also known as tumor-specific active immunotherapy, are a type of tumor immunotherapy. Their principle involves using tumor cells and tumor antigens to prepare vaccines in various forms, which are then injected into cancer patients. Antigen-presenting cells (APCs) take up the vaccines and present them to immune cells, such as T cells. T-cell-based immunotherapy targets peptide epitopes derived from tumor-associated or tumor-specific proteins presented by the major histocompatibility complex (MHC). The antigens recognized by tumor-specific T lymphocytes, i.e., their epitopes, can be molecules derived from all protein types, such as enzymes, receptors, and transcription factors. These are expressed in the cells of the corresponding tumor, and their expression is usually upregulated compared to homologous unaffected cells, thereby activating T lymphocytes and producing cytotoxic T lymphocytes (CTLs). Specific binding to and killing of tumor cells is one of the hot topics in tumor immunology research in recent years.

[0004] Currently, tumor vaccine development typically targets two types of antigens: tumor-associated antigens (TAAs) and tumor-specific antigens (TSAs). TAAs directly participate in the carcinogenic process, exhibiting high or characteristic expression in various tumor cells, primarily derived from gene amplification or post-translational modification. Targeted therapy against TAAs is a crucial approach to cancer treatment. TAA-derived peptide vaccines can effectively kill different types of tumor cells carrying the same tumor antigen (Shimizu Y et al., Cancer Sci 2018). Effective TAAs can inhibit cancer cell proliferation and survival; using such TAAs as targets for immunotherapy can minimize the risk of cancer cell immune escape. Cancer cell immune escape can be attributed to TAA deletion, mutation, or downregulation during cancer cell proliferation. Therefore, the identification of novel TAAs capable of inducing efficient and specific tumor immune responses is crucial for tumor vaccine development (Vansteenkiste JF et al., 2016; Shimizu Y et al., 2018).

[0005] Studies have found overlap in gene expression between induced pluripotent stem cells (iPSCs), embryonic stem cells (ESCs), and tumor cells, with both expressing some tumor antigens. These tumor antigens can induce a specific immune response against the tumor, becoming effective targets for activating anti-tumor immune responses. Recent studies have further confirmed the application prospects of stem cell-derived tumor antigens as effective tumor vaccines. Using autologous iPSCs as a prophylactic tumor vaccine can effectively inhibit the growth of various cancers (melanoma, breast cancer, and mesenchymal tumors) and generate a specific immune response against tumor antigens (Kooreman NG et al., Cell Stem Cell 2018). Tumor-associated antigens (TAAs) derived from embryonic stem cells have been tested in preclinical studies, such as alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and cancer-testis antigens (CTAs). These peptide vaccines can effectively eliminate tumor cells expressing these antigens (Brichard VG et al., 2007; Atanackovic D et al., 2008). A recently discovered embryonic antigen, clandin 6 (CLDN6), is associated with various tumorigenesis processes. CLDN6-CAR-T cells, prepared using CLDN6 as a target protein captured by a chimeric antigen receptor (CAR), can effectively inhibit the growth of solid tumors such as colon cancer, ovarian cancer, and lung cancer (Reinhard Ke et al., Science 2020). Therefore, by analyzing and screening tumor antigens that are co-expressed highly among induced pluripotent stem cells, embryonic stem cells, and tumor cells, and analyzing the antigenic epitope peptides expressed on these antigens, highly effective tumor vaccines targeting various tumors can be prepared.

[0006] References:

[0007] (1)Brichard VG,Lejeune D.GSK's antigen-specific cancer immunotherapy program:pilot results leading to Phase III clinical development.Vaccine2007,25Suppl 2:B61-71.

[0008] (2)Atanackovic D,Altorki NK,Cao Y,Ritter E,Ferrara CA,Ritter G,Hoffman EW,Bokemeyer C,Old LJ,Gnjatic S.Booster vaccination of cancerpatients with MAGE-A3 protein reveals long-term immunological memory ortolerance depending on priming.Proc Natl Acad Sci U S A 2008,105(5):1650-1655.

[0009] (3)Vansteenkiste JF,Cho BC,Vanakesa T,De Pas T,Zielinski M,Kim MS,Jassem J,Yoshimura M,Dahabreh J,Nakayama H,Havel L,Kondo H,Mitsudomi T,Zarogoulidis K,Gladkov OA,Udud K,Tada H,Hoffman H,Bugge A,Taylor P,GonzalezEE,Liao ML,He J,Pujol JL,Louahed J,Debois M,Brichard V,Debruyne C,Therasse P,Altorki N.Efficacy of the MAGE-A3 cancer immunotherapeutic as adjuvanttherapy in patients with resected MAGE-A3-positive non-small-cell lung cancer(MAGRIT):a randomised,double-blind,placebo-controlled,phase 3trial.LancetOncol 2016,17(6):822-835.

[0010] (4)Shimizu Y, Suzuki T, Yoshikawa T, Tsuchiya N, Sawada Y, Endo I, Nakatsura T. Cancer immunotherapy-targeted glypican-3or neoantigens. Cancer Sci2018,109(3):531-541.

[0011] (5)Kooreman NG, Kim Y, de Almeida PE, Termglinchan V, Diecke S, Shao NY, Wei TT, Yi H, Dey D, Nelakanti R, Brouwer TP, Paik DT, Sagiv-Barfi I, Han A, QuaxPHA, Hamming JF, Levy R, Davis MM, Wu JC. Autologous iPSC-Based Vaccines ElicitAnti-tumor Responses In Vivo.Cell Stem Cell 2018,22(4):501-513e507.

[0012] (6)Reinhard K,Rengstl B,Oehm P,Michel K,Billmeier A,Hayduk N,Klein O,Kuna K,Ouchan Y,Woll S,Christ E,Weber D,Suchan M,Bukur T,Birtel M,Jahndel V,Mroz K,Hobohm K,Kranz L,Diken M,Kuhlcke K,Tureci O,Sahin U.An RNA vaccine drives expansion and efficacy of claudin-CAR-T cells against solidtumors.Science 2020,367(6476):446-453. Summary of the Invention

[0013] To address the issues of weak immunogenicity of tumor antigens and immune escape of tumor cells during tumor treatment, this invention provides tumor antigen epitope peptides derived from embryonic stem cells and their applications.

[0014] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0015] On one hand, the present invention provides a tumor antigen epitope peptide, wherein the tumor antigen epitope peptide comprises one or more sequences selected from those shown in SEQ ID NO:1-10.

[0016] In another aspect, the present invention provides the use of the above-mentioned tumor antigen epitope peptide in the preparation of antitumor drugs and / or vaccines.

[0017] Preferably, the antitumor drug and / or vaccine further includes an immune adjuvant.

[0018] Preferably, the immune adjuvant comprises CpG and / or Poly IC.

[0019] In another aspect, the present invention provides an antitumor pharmaceutical composition comprising the above-mentioned tumor antigen epitope peptide.

[0020] Preferably, the antitumor drug composition further includes a pharmaceutically acceptable carrier.

[0021] In the technical solution of the present invention, the tumor includes bladder cancer, esophageal cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, stomach cancer, and uterine cancer; preferably bladder cancer.

[0022] The above technical solution has the following advantages or beneficial effects:

[0023] This invention employs a combination of theoretical and experimental methods to screen tumor antigens expressed by ESCs and identify CTL epitope peptides that effectively induce specific tumor immune responses. Multiple different epitope peptides are then applied together to tumor cells to avoid off-target effects and tumor cell immune escape during tumor treatment.

[0024] This invention performed transcriptome sequencing analysis on mouse MB49 cell line and embryonic stem cells, selecting 100 genes (top 100 in signal-to-noise ratio) from over 2000 cancer-related genes that tend to be highly expressed in iPSC, ESC, and MB49 cell lines, but lowly expressed or not expressed in normal tissues. Then, 59 relatively highly expressed genes were screened out. Through software predictive analysis, four potentially effective tumor antigens were identified: epidermal cell transformation sequence 2 (ECT2), actin-binding protein (ANLN), maternal embryoleucine zipper kinase (MELK), and cyclin B1 (CCNB1), along with 10 corresponding epitope peptides. These proteins are highly expressed in bladder cancer and various other tumors, but expressed at low levels in normal tissues. When the body generates specific T cells targeting these peptides, these cells can attack and kill tumor cells containing the same peptides. This invention further validated the inhibitory effect of five epitope peptides on tumor growth in a bladder cancer tumor model through animal experiments, demonstrating that mice treated with the peptides produced specific T lymphocytes that secrete IFN-γ.

[0025] The tumor antigens and their epitope peptides screened by this invention can effectively improve the screening efficiency of tumor antigens and their epitope peptides. After verification, the screened epitope peptides all have a good inhibitory effect on tumor growth. Attached Figure Description

[0026] Figure 1 This is a graph showing the expression analysis of tumor-expressing genes screened in Example 2 in various tumor cells.

[0027] Figure 2 This is a graph showing the test results of the inhibitory effect of ESC on tumor growth in Example 3.

[0028] Figure 3 This is a graph showing the immunogenicity test results of the peptide in Example 4.

[0029] Figure 4 This is a graph showing the test results of the inhibitory effect of the peptide in Example 5 on tumor growth.

[0030] Figures 5a-5e This is a graph showing the test results of the ability of the peptide in Example 6 to stimulate a T-cell immune response. Detailed Implementation

[0031] The following embodiments are merely some, not all, of the embodiments of the present invention. Therefore, the detailed descriptions of the embodiments provided below are not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0032] In this invention, unless otherwise specified, all equipment and raw materials are commercially available or commonly used in the industry. The methods described in the following embodiments are conventional methods in the art, unless otherwise specified.

[0033] Example 1: Gene Screening

[0034] Transcriptome sequencing and analysis of mouse tumor cells and embryonic stem cells revealed genes that are highly expressed in both tumor cells and embryonic stem cells. Based on literature review and in vitro experimental verification, effective genes were initially identified, including four genes: ECT2, ANLN, CCNB1, and MELK. The antigenic epitopes of these genes were also obtained through analysis. In addition, the antigenic epitope peptides and elongation peptides of these genes were synthesized in collaboration with Jier Biochemical (Shanghai) Co., Ltd.

[0035] The sequences of the above proteins and their epitope peptides are shown in Table 1.

[0036] Table 1

[0037]

[0038]

[0039] In Table 1, the elongated peptide contains the epitope peptide sequence and functions the same as the epitope peptide. Because dendritic cells require a cleavage process to present antigenic epitopes, the actual synthesis involves an elongated peptide containing the epitope peptide. Population coverage refers to the proportion of the population possessing at least one HLA allele that can bind to the peptide, calculated using the method described in http: / / tools.iedb.org / population / based on the frequency of each HLA allele in the population. HLA alleles are part of the human major histocompatibility complex (MHC). Based on the characteristics of the encoding molecules, the genes of the entire complex can be divided into three classes: class I, class II, and class III genes.

[0040] Example 2: Gene expression in normal and tumor tissues:

[0041] Transcriptome sequencing and analysis of mouse tumor cells and normal tissues identified differentially expressed genes in tumor and normal tissues, as shown in the following results. Figure 1 As shown, the expression levels of ECT2, ANLN, CCNB1, and MELK genes are high in various tumor tissues but low in normal tissues, including the bladder, esophagus, kidney, liver, lung, ovary, pancreas, prostate, stomach, and uterus. This suggests that epitope peptides derived from these genes have the potential to serve as tumor antigens.

[0042] Example 3: Inhibitory effect of two mouse embryonic stem cell vaccines on bladder cancer:

[0043] In this embodiment, 4-6 week old C57B / 6J mice were used as the experimental model.

[0044] The specific experimental protocol was as follows: On Day 0, each mouse was subcutaneously inoculated with 5^10 5 MB49 bladder cancer cells, injection of 2^10 starting on Day 10. 6 Irradiated ESC (C57B / 6J source) or ESC (129 source) vaccine, or ESC vaccine plus CPG adjuvant plus GM-CSF, were administered to each animal at a total dose of 100 μL. The amount of CPG was 1 μM and the amount of GM-CSF was 10 μg per animal. Immunization was performed weekly for a total of four times. The negative control was immunized with the same amount of PBS, with each animal receiving a total dose of 100 μL. The experimental animals were euthanized five days after the fourth immunization, and tumor size was measured twice a week.

[0045] In this embodiment, the inhibitory effects of different injections on tumor growth are shown in [the following text is missing]. Figure 2 :

[0046] in, Figure 2 The top part is a schematic diagram of the mouse tumor inoculation and immunization protocol;

[0047] Figure 2 The middle section shows the inhibitory effect of the ESC (C57B / 6J-derived) vaccine on mouse tumor growth. The left side shows a comparison of the actual tumor size of mice in different groups, and the right side shows the tumor growth curve of mice. The horizontal axis represents the growth cycle of mice after tumor implantation, and the vertical axis represents the tumor volume of mice. The data are expressed as mean ± SD. As can be seen from the figure, the ESC (C57B / 6J-derived) vaccine can significantly inhibit the growth of tumor tissue compared with the control group injected with PBS.

[0048] Figure 2The bottom part shows the effect of ESC(129-derived) vaccine on the inhibition of mouse tumor growth. The left side shows the comparison of the actual size of the tumors in different groups of mice, and the right side shows the tumor growth curve of mice. The horizontal axis is the growth cycle after tumor implantation in mice, and the vertical axis is the tumor volume in mice. The data are expressed as mean ± SD. As can be seen from the figure, the combination of ESC(129) and CPG has a certain tumor inhibition effect. However, the combined effect of ESC(129-derived) vaccine, adjuvant CPG and immunomodulatory factor GM-CSF has a better tumor inhibition effect than the CPG+GM-CSF group and the ESC(129)+CPG group.

[0049] Example 4: Immunogenicity identification of epitope peptide extension peptide:

[0050] In this embodiment, 4-6 week old C57B / 6J mice were used as the experimental model.

[0051] The specific experimental protocol was as follows: Each mouse in the experimental group was subcutaneously injected with ESC vaccine, or with ESC vaccine plus CPG adjuvant plus GM-CSF, with a total injection volume of 100 μl per mouse. The amount of CPG was 1 μM, and the amount of GM-CSF was 10 μg per mouse. Immunization was repeated every 7 days, for a total of two immunizations. The negative control group was immunized with the same amount of PBS, with a total injection volume of 100 μl per mouse. Five days after the second immunization, the experimental animals were euthanized, and the spleen tissue was aseptically removed, ground, and monocytes were separated using lymphocyte separation medium. Monocytes were incubated in ELISPOT plate wells and stimulated with extended peptides of the selected epitope peptides: Ccnb1peptide (SEQ ID NO:11), Anln peptide (SEQ ID NO:14), Ect2-1 peptide (SEQ ID NO:16), Ect2-2 peptide (SEQ ID NO:18), and Melk peptide (SEQ ID NO:12). The cells were incubated at 37°C for 20 hours, and spot formation was detected to determine the number of T cells specifically secreting IFN-γ in the spleen cells of immunized mice.

[0052] In this embodiment, the immunogenicity test results of the peptide are shown in the figure. Figure 3 :

[0053] in, Figure 3 The left side shows a dot plot of IFN-γ-specific secretion by mouse T cells after immunization with ESC vaccine stimulated by different peptides; the right side shows the statistical results, where the horizontal axis represents different peptide types, the vertical axis represents the number of T cells that specifically secrete IFN-γ, and the data are expressed as mean ± SD; SFU represents dot-forming unit; it can be seen from the figure that mice immunized by ESC can produce tumor-specific T cells that secrete IFN-γ in response to the selected peptides.

[0054] Example 5: Inhibitory effect of tumor antigen epitope peptides on bladder cancer:

[0055] In this embodiment, 4-6 week old C57B / 6J mice were used as the experimental model.

[0056] The specific protocol involved subcutaneously injecting 5*10 mg of the experimental mice on Day 0. 5 MB49 bladder cancer cells were immunized with peptides on days 3, 10, and 14, and the animals were euthanized on day 18. Each mouse in the experimental group was subcutaneously injected with an extended peptide of a synthetic epitope peptide plus adjuvant CPG, with a total injection volume of 100 μL per mouse. The CPG concentration was 1 μM. The peptide immunizations were divided into: CCNB1 (corresponding to SEQ ID NO:11), MELK (corresponding to SEQ ID NO:12) + ANLN (corresponding to SEQ ID NO:14), and ECT2-1 (corresponding to SEQ ID NO:16) + ECT2-2 (corresponding to SEQ ID NO:18). Mice in the control group were immunized with the same amount of PBS, with a total injection volume of 100 μL per mouse. Tumor size was measured twice a week.

[0057] In this embodiment, the test results of the inhibitory effect of the peptide on tumor growth are shown in the figure. Figure 4 :

[0058] Figure 4 The top figure is a schematic diagram of mouse tumor inoculation and mouse immunization protocols; the bottom figure is a mouse tumor growth curve, with the horizontal axis representing the growth cycle after tumor inoculation and the vertical axis representing the tumor volume. The data are expressed as mean ± SD. As can be seen from the figure, compared with the control group PBS, the tumor growth of mice injected with extended peptide plus adjuvant was inhibited. Among them, the combination of MELK and ANLN treatment showed the best effect.

[0059] Example 6: Detection of the ability of specific T cells to secrete IFN-γ induced by tumor antigen epitope peptides:

[0060] The spleen tissue of the euthanized experimental animals from Example 5 was aseptically removed, ground, and then monocytes were isolated using lymphocyte separation medium. The monocytes were incubated in ELISPOT plate wells and simultaneously stimulated. The cells were cultured at 37°C and cytokine captured. After 20 hours, spot formation was detected, and the number of T cells specifically secreting IFN-γ in the spleen cells of immunized mice was determined.

[0061] In this embodiment, the test results of the peptide's ability to stimulate a T-cell immune response are shown in the figure. Figures 5a-5e :

[0062] Figures 5a-5eThe left image shows a dot plot of IFN-γ-specific secretion by mouse T cells after peptide-stimulated immunization with peptide vaccine; the right image shows the statistical results, where the horizontal axis represents different experimental groups, the vertical axis represents the number of T cells that specifically secrete IFN-γ, and the data are expressed as mean ± SD; SFU represents dot-forming unit; it can be seen from the figure that the number of T cells secreting IFN-γ increased significantly after stimulation with the five elongation peptides.

[0063] In summary, the tumor antigens and their epitope peptides screened by this invention can effectively improve the screening efficiency of tumor antigens and their epitope peptides. Verification has shown that the screened epitope peptides all have a good inhibitory effect on tumor growth.

[0064] The above description is merely a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent transformations made based on the content of the present invention's specification and drawings, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention. SEQUENCE LISTING <110> Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences <120> Tumor antigen epitope peptides derived from embryonic stem cells and their applications <130> 2021 <160> 20 <170> PatentIn version 3.3 <210> 1 <211> 9 <212> PRT <213> Artificial synthesis <400> 1 Tyr Val Lys Asp Ile Tyr Ala Tyr Leu 1 5 <210> 2 <211> 9 <212> PRT <213> Artificial synthesis <400> 2 Ser Ser Ile Leu Leu Leu Gln Gln Met 1 5 <210> 3 <211> 9 <212> PRT <213> Artificial synthesis <400> 3 His Tyr Asn Val Thr Thr Thr Arg Leu 1 5 <210> 4 <211> 9 <212> PRT <213> Artificial synthesis <400> 4 Ser Ser Met Ser Leu Leu Ala Pro Leu 1 5 <210> 5 <211> 9 <212> PRT <213> Artificial synthesis <400> 5 Val Ile Tyr Gln Ala Ser Gln Ala Leu 1 5 <210> 6 <211> 9 <212> PRT <213> Artificial synthesis <400> 6 Val Ala Val Ser Leu Gly Thr Pro Ile 1 5 <210> 7 <211> 9 <212> PRT <213> Artificial synthesis <400> 7 Asn Tyr Val Asn Ile Leu Ala Thr Ile 1 5 <210> 8 <211> 9 <212> PRT <213> Artificial synthesis <400> 8 Lys Thr Tyr Pro Pro Phe Val Asn Phe 1 5 <210> 9 <211> 9 <212> PRT <213> Artificial synthesis <400> 9 Arg Leu Pro Ser Val Ala Leu Leu Leu 1 5 <210> 10 <211> 9 <212> PRT <213> Synthetic <400> 10 Thr Tyr Pro Pro Phe Val Asn Phe Phe 1 5 <210> 11 <211> 25 <212> PRT <213> Synthetic <400> 11 Ala Asp Pro Asn Leu Cys Ser Glu Tyr Val Lys Asp Ile Tyr Ala Tyr 1 5 10 15 Leu Arg Gln Leu Glu Glu Glu Gln Ala 20 25 <210> 12 <211> 25 <212> PRT <213> Synthetic <400> 12 Asp Val Pro Lys Trp Leu Ser Pro Ser Ser Ile Leu Leu Leu Gln Gln 1 5 10 15 Met Leu Gln Val Asp Pro Lys Lys Arg 20 25 <210> 13 <211> 25 <212> PRT <213> Synthetic <400> 13 Asp Gly Pro Arg Arg Leu Lys Leu His Tyr Asn Val Thr Thr Thr Arg 1 5 10 15 Leu Val Asn Pro Asp Gln Leu Leu Asn 20 25 <210> 14 <211> 25 <212> PRT <213> Artificial synthesis <400> 14 Glu Gln Glu Asp Ala Leu Asn Ile Ser Ser Met Ser Leu Leu Ala Pro 1 5 10 15 Leu Ala Gln Thr Val Gly Val Val Ser 20 25 <210> 15 <211> 25 <212> PRT <213> Artificial synthesis <400> 15 Asn Glu Ile Asn Met Gln Gln Thr Val Ile Tyr Gln Ala Ser Gln Ala 1 5 10 15 Leu Asn Cys Cys Val Asp Glu Glu His 20 25 <210> 16 <211> 25 <212> PRT <213> Artificial synthesis <400> 16 Cys Thr Gln Gly Glu Lys Phe Arg Val Ala Val Ser Leu Gly Thr Pro 1 5 10 15 Ile Met Lys Pro Glu Trp Ile Tyr Lys 20 25 <210> 17 <211> 25 <212> PRT <213> Artificial synthesis <400> 17 Lys Glu Leu Tyr Gln Thr Glu Ser Asn Tyr Val Asn Ile Leu Ala Thr 1 5 10 15 Ile Ile Gln Leu Phe Gln Val Pro Leu 20 25 <210> 18 <211> 25 <212> PRT <213> Synthetic <400> 18 Leu Lys Tyr Ser Lys Asp Leu Val Lys Thr Tyr Pro Pro Phe Val Asn 1 5 10 15 Phe Phe Glu Met Ser Lys Glu Thr Ile 20 25 <210> 19 <211> 25 <212> PRT <213> Synthetic <400> 19 Glu Leu Leu Ile Arg Pro Val Gln Arg Leu Pro Ser Val Ala Leu Leu 1 5 10 15 Leu Asn Asp Leu Lys Lys His Thr Ala 20 25 <​​​​​​​​​​​​​​​​

Claims

1. The use of tumor antigen epitope peptides in the preparation of antitumor drugs and / or vaccines, characterized in that, The sequence of the tumor antigen epitope peptide is shown in SEQ ID NO:11; the tumor is bladder cancer.

2. The use according to claim 1, characterized in that, The antitumor drugs and / or vaccines also include immune adjuvants.

3. The use according to claim 2, characterized in that, The immune adjuvants include CpG and / or Poly IC.

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