Pharmaceutical composition containing 11-HETE and application of pharmaceutical composition in preparation of anti-tumor or auxiliary anti-tumor drugs

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a pharmaceutical composition containing 11-HETE and application of the pharmaceutical composition in preparation of anti-tumor or auxiliary anti-tumor drugs. It is found for the first time that exogenous supplement of 11-HETE including (R) 11-HETE, (S) 11-HETE or (R / S) 11-HETE mixture can target CD8 + T cells, and the infiltration, proliferation and killing functions of the CD8 + T cells are enhanced. The 11-HETE can directly promote immune surveillance and inhibit tumor growth, and can sensitize anti-tumor treatments such as chemotherapy, immunotherapy, chemotherapy-immunotherapy combined therapy, CAR-T cell therapy and the like. Therefore, the pharmaceutical composition containing the 11-HETE and the application of the pharmaceutical composition in preparation of anti-tumor drugs or tumor adjuvant therapy and the like have wide clinical application prospects and commercial development values.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a pharmaceutical composition containing 11-HETE and use thereof in the preparation of an anti-tumor or adjuvant anti-tumor drug. BACKGROUND

[0002] Tumor is a major disease that seriously threatens human life and health, and its occurrence and development not only depends on the genetic and epigenetic abnormalities of tumor cells themselves, but also is deeply affected by a variety of non-tumor cell components in the tumor microenvironment and their interactions. In recent years, the role of the tumor microenvironment in determining tumor treatment response and prognosis has gradually been valued, and tumor treatment strategies have gradually developed from single targeting of tumor cells to comprehensive regulation of the tumor microenvironment.

[0003] At present, chemotherapy, immune checkpoint inhibitor therapy, and chimeric antigen receptor T cell (CAR-T) therapy have become important treatment methods for various solid tumors and hematological tumors. For example, in bladder cancer, neoadjuvant chemotherapy represented by gemcitabine combined with cisplatin (GP regimen) is a commonly used first-line treatment regimen in clinical practice; in colorectal cancer, bladder cancer and other tumors, immunotherapy represented by anti-PD-1 antibody has also shown certain efficacy; CAR-T cell therapy as a new generation of cell therapy technology is also expanding to the field of solid tumors. However, the above treatment methods generally face large differences in therapeutic effect, treatment tolerance, and non-response in some patients in clinical application, which limits their overall therapeutic effect.

[0004] A large number of studies have shown that whether it is chemotherapy, immunotherapy or CAR-T cell therapy, its anti-tumor effect depends to a large extent on the activation state of the body's anti-tumor immune response, especially the cytotoxic immune function mediated by CD8 + In solid tumors, the tumor microenvironment often inhibits the infiltration, proliferation and killing activity of CD8 + T cells through a variety of mechanisms, thereby weakening the anti-tumor treatment effect. Therefore, how to enhance the anti-tumor immune response through combined drug use or auxiliary intervention on the basis of existing treatment regimens is an important research direction in the field of tumor treatment.

[0005] Cancer-associated fibroblasts (CAFs) are one of the most abundant stromal cells in the tumor microenvironment, and their roles in tumor development and treatment response have attracted increasing attention. Previous studies have considered CAFs as pro-tumor factors, but as research deepens, more and more evidence shows that CAFs have significant functional heterogeneity and may participate in regulating anti-tumor immune response and affecting treatment effect under certain conditions. However, whether CAFs undergo metabolic and functional remodeling during anti-tumor therapy and how they affect immune cell function through which molecular mediators are still lacking clear conclusions.

[0006] In recent years, the role of lipid metabolism and its oxidation products in the tumor microenvironment has gradually become a research hotspot. Arachidonic acid is an important precursor of various bioactive lipids, and its oxidation can generate various oxidized fatty acids. Existing researches mainly focus on classic lipid mediators such as prostaglandins and leukotrienes, but the role of other oxidized fatty acids in tumor immune regulation is still limited. Different oxidized fatty acids may have different or even opposite biological effects in different cell types and disease backgrounds, and this complexity brings challenges to their application in tumor therapy.

[0007] Arachidonic acid (AA) is the main precursor substrate for the biosynthesis of numerous eicosanoids in tumors. However, the role of AA and its metabolites in tumor therapy is controversial: some studies have shown that it has pro-inflammatory and pro-tumor progression effects; there is also evidence that certain AA metabolites can induce apoptosis or inhibit angiogenesis. This contradiction suggests that the function of the AA metabolic network is highly context-dependent, and its final effect may depend on specific metabolic enzymes, specific eicosanoid species produced, and specific cell types affected. This complexity leads to uncertainty in the direction of therapeutic strategies targeting the AA pathway, making it difficult to design precisely.

[0008] 11-hydroxyeicosatetraenoic acid (11-HETE) is one of the important products of arachidonic acid oxidation metabolism, with a clear chemical structure, including (R) 11-HETE and (S) 11-HETE, and other stereoisomer forms. Existing literature has reported the role of 11-HETE in inflammatory response and related physiological processes, but its application value in the context of tumor therapy, especially in combination with chemotherapy, immunotherapy or CAR-T cell therapy, has not been systematically studied. Whether different stereoisomers of 11-HETE play different biological roles in the tumor microenvironment, and whether it can be used as an adjunct to existing anti-tumor treatment regimens to improve their efficacy, there is still a lack of clear technical solutions. SUMMARY

[0009] In order to overcome the defects and deficiencies in the prior art, the present application provides a pharmaceutical composition containing 11-HETE and its use in the preparation of an anti-tumor or adjuvant anti-tumor drug.

[0010] In particular, the present application provides an anti-tumor or adjuvant therapy drug based on 11-HETE (including (R) 11-HETE, (S) 11-HETE, or (R / S) 11-HETE mixture), especially an anti-tumor drug targeting chemotherapy, immunotherapy, chemo-immunotherapy, and CAR-T cell therapy for bladder cancer.

[0011] In order to achieve the above-mentioned application purposes, the present application adopts the following technical solutions: In a first aspect, the present application provides a pharmaceutical composition having an anti-tumor effect or an adjuvant anti-tumor effect, which contains 11-hydroxyeicosatetraenoic acid (11-HETE) as the only active ingredient or contains 11-HETE and other anti-tumor drugs.

[0012] As an optional mode, in the above-mentioned pharmaceutical composition, the 11-HETE is selected from one or more of (R) 11-HETE, (S) 11-HETE, or (R / S) 11-HETE mixture.

[0013] As an optional mode, in the above-mentioned pharmaceutical composition, the 11-HETE targets CD8 + T cells, enhances CD8 + T cell infiltration, proliferation, and killing function, directly promotes immune surveillance to inhibit tumor growth, and has a sensitizing effect on the other anti-tumor drugs.

[0014] As an optional mode, in the above-mentioned pharmaceutical composition, the other anti-tumor drugs include chemotherapy drugs or immune anti-tumor drugs.

[0015] As an optional mode, in the above-mentioned pharmaceutical composition, the chemotherapy drugs include one or more of alkylating agents, antimetabolites, anti-tumor antibiotics, plant alkaloids, or platinum anti-tumor drugs, and the immune anti-tumor drugs include immune checkpoint inhibitors or CAR-T cell therapy.

[0016] As an optional mode, in the above-mentioned pharmaceutical composition, the chemotherapy drugs are selected from one or more of capecitabine, gemcitabine, paclitaxel, docetaxel, cyclophosphamide, cisplatin, carboplatin, oxaliplatin, or 5-FU, and the immune checkpoint inhibitors are selected from one or more of PD-1 inhibitors, PD-L1 inhibitors, or CTLA-4.

[0017] Preferably, the chemotherapeutic drug is a combination of gemcitabine and cisplatin (i.e., GP chemotherapy regimen).

[0018] Preferably, the immune checkpoint inhibitor is a PD-1 antibody or a PD-L1 antibody.

[0019] In a second aspect, the present application provides use of the pharmaceutical composition of the first aspect described above in the manufacture of an anti-tumor or an adjuvant anti-tumor drug.

[0020] As an optional mode, in the use described above, the drug further comprises a pharmaceutically acceptable excipient.

[0021] As an optional mode, in the use described above, the drug of the present application can be used in various administration modes, including bladder local administration / infusion or oral, intravenous administration or CAR-T pretreatment, etc.

[0022] As an optional mode, in the use described above, the drug is an oral preparation or a parenteral preparation.

[0023] As an optional mode, in the use described above, the various active ingredients in the drug can be in the same preparation or in different preparations.

[0024] As an optional mode, in the use described above, the various active ingredients in the drug can be administered simultaneously or not simultaneously.

[0025] As an optional mode, in the use described above, the tumor is a solid tumor.

[0026] As an optional mode, in the use described above, the tumor is selected from one or more of the following: bladder cancer, colon cancer, prostate cancer, glioblastoma, endometrial cancer, breast cancer, colorectal cancer, ovarian cancer, melanoma, head and neck squamous cell carcinoma or non-small cell lung cancer.

[0027] Preferably, the tumor is bladder cancer or colorectal cancer.

[0028] Preferably, in the use described above, the drug of the present application is used in the following anti-tumor treatment regimen: 11-HETE alone for treating bladder cancer; 11-HETE in combination with GP chemotherapy regimen for treating bladder cancer; 11-HETE in combination with anti-PD1 immunotherapy for treating bladder cancer; 11-HETE, GP chemotherapy regimen and anti-PD1 immunotherapy for treating bladder cancer; 11-HETE combined with CAR-T cell therapy for treating bladder cancer; and 11-HETE combined with anti-PD1 immunotherapy for treating colorectal cancer.

[0029] Compared with the prior art, the present application has the following beneficial effects: The present application has been verified by in vitro and in vivo experiments that CD8 + T cell infiltration, proliferation and killing function in the tumor microenvironment are inhibited by the activity of PTEN enzyme in T cells, thereby limiting the efficacy of tumor chemotherapy, immunotherapy, combined chemotherapy and immunotherapy, and CAR-T therapy. + The present application first found that exogenous supplement of 11-HETE (including (R) 11-HETE, (S) 11-HETE, and (R / S) 11-HETE mixture) can target CD8 + T cells, enhance CD8 + T cell infiltration, proliferation and killing function. 11-HETE can directly promote immune surveillance to inhibit tumor growth, and can enhance the sensitivity of chemical drug treatment, immunotherapy, combined chemotherapy and immunotherapy, and CAR-T cell therapy, etc. Anti-tumor treatment. Therefore, the pharmaceutical composition comprising 11-HETE and its use in the preparation of anti-tumor drugs provided by the present application have broad clinical application prospects and commercial development value. BRIEF DESCRIPTION OF DRAWINGS

[0030] The accompanying drawings are included to provide a further understanding of the present application, and constitute a part of the specification, together with the embodiments of the present application, to explain the present application, and do not constitute a limitation of the present application. In the drawings: Figure 1 : In in vivo experiments, (R / S) 11-HETE mixture and its separated chiral substances (R) 11-HETE and (S) 11-HETE single drug control bladder tumor growth and enhance CD8 + T cell infiltration and killing function in the tumor microenvironment.

[0031] Figure 2 : In in vitro experiments, (R / S) 11-HETE mixture promotes CAR-T cell killing of bladder tumor; in in vivo experiments, (R / S) 11-HETE mixture combined with CAR-T cell therapy controls bladder tumor growth.

[0032] Figure 3 : In in vivo experiments, (R / S) 11-HETE mixture improves the response rate of bladder tumor chemotherapy, and prolongs the survival period of tumor-bearing mice with bladder tumor treated by chemotherapy.

[0033] Figure 4In vivo, (R / S) 11-HETE mixture combined with immunotherapy controlled the growth of bladder tumor and enhanced CD8 + T cell infiltration and killing function.

[0034] Figure 5 In vivo, (R / S) 11-HETE mixture combined with immunotherapy controlled the growth of colon tumor and enhanced CD8 + T cell infiltration and killing function.

[0035] Figure 6 In vivo, (R / S) 11-HETE mixture further enhanced the inhibitory effect of chemotherapy combined with immunotherapy on the growth of bladder tumor. DETAILED DESCRIPTION

[0036] The present application will be further described below with reference to specific examples. It should be understood that the specific examples described herein are intended to explain the present application and are not intended to limit the scope of the present application.

[0037] Example: Unless otherwise specified, the techniques or conditions in the examples were carried out according to the techniques or conditions described in the literature in the art, or according to the product instructions. Unless otherwise specified, the reagents or instruments used were conventional products that can be purchased through regular channels.

[0038] Unless otherwise specified, the experimental methods in the following examples were conventional methods. Unless otherwise specified, the experimental materials used in the following examples were commercially available products.

[0039] 11-HETE (including (R) 11-HETE, (S) 11-HETE or (R / S) 11-HETE mixture) can be purchased through commercial channels.

[0040] In the following experiments, (R / S) 11-HETE mixture can also be represented as (±) 11-HETE. In addition, (R) 11-HETE, (S) 11-HETE or (R / S) 11-HETE mixture can be used interchangeably with 11(R)-HETE, 11(S)-HETE or 11(R / S)-HETE mixture, respectively, all of which represent the same meaning.

[0041] 1. Experimental methods In vivo experiments, C57BL / 6 mice were purchased from Guangdong Medical Laboratory Animal Center (Foshan, China), NCG mice were purchased from GemPharmatech (Jiangsu, China). In subcutaneous tumor model, C57BL / 6, NCG mice were inoculated with the indicated tumor cells subcutaneously in the right flank, and the mice were sacrificed when the tumors reached the preset size for subsequent analysis; for mice that need further treatment, the mice were randomly divided into groups based on similar tumor volume and body weight, and then given subsequent drug or treatment. In adoptive transfer experiment, the in vitro activated CAR-CD8 + T cells were transferred to tumor-bearing mice. In N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)-induced spontaneous bladder cancer model, 6-8-week-old wild-type C57BL / 6 mice were used, 0.05% BBN was added to the drinking water for free drinking, and high-resolution ultrasound (HR-Ultrasound) was used to monitor tumor occurrence and carry out treatment research based on the monitoring. In orthotopic bladder tumor model, 1x10 6 MB49 luciferase Cells were inoculated into the bladder of mice and retained for 1 h, and subsequent in vivo experiments were carried out according to the random principle after inoculation.

[0042] Cell lines used: SYBC1, UMUC3, T24, MC38 and MB49 cells were cultured in DMEM or RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS). All cell lines were identified by STR typing and confirmed to be free of mycoplasma contamination.

[0043] CAR-T cell cytotoxicity experiment: activated CD8 + T cells were inoculated in culture plates coated with RetroNectin, and then HER2-CAR lentivirus was added to the culture plates; 8 h later, the culture medium was replaced, and the culture was continued for 72 h. The obtained HER2-CAR-expressed CD8 + T cells were evaluated by flow cytometry. Tumor cells were inoculated in 6-well plates, and CAR-T cells were added at an effector to target (E:T) ratio of 2:1 after the cells adhered, and incubated at 37°C for 24 h; then all cells were collected and further analyzed by flow cytometry.

[0044] LDH release experiment: CytoTox 96 Non-Radioactive Cytotoxicity Assay Kit (Promega) was used according to the instructions to detect the activity of lactate dehydrogenase (LDH) in the cell culture supernatant under different treatment conditions, to evaluate the level of pyroptosis.

[0045] Flow cytometry: For cell surface marker analysis, cells were resuspended in PBS containing 1% FBS and stained with fluorescent conjugated antibodies against CD45, CD3, CD8 at 4°C for 30 min. For intracellular cytokine staining, cells were incubated with PMA and ionomycin (eBioscience) at 37°C for 4 h with shaking; then fixed and permeabilized according to the intracellular staining kit (eBioscience) instruction and stained with anti-IFN-γ and TNF-a antibodies. All samples were detected and analyzed on a CytoFLEX LX flow cytometer.

[0046] Apoptosis analysis: Cells treated with indicated chemotherapy drugs were digested and separated with 0.25% trypsin (without EDTA) and collected by centrifugation. Cells were stained and detected according to the kit instruction using the apoptosis detection kit (BioLegend).

[0047] Quantification and statistical analysis: Cell experiments were performed in biological triplicates unless otherwise specified. Statistical analysis was completed using GraphPad Prism 9.0 or SPSS 24 software. Student’s t-test was used for comparison between two groups (treatment group vs. control group); one-way ANOVA was used for comparison of continuous variables among multiple groups. Survival analysis was estimated by the Kaplan-Meier method. Unless otherwise stated, statistical details (including the statistical test method used, the exact number of sample size n, and the precision index (mean ± SD or SEM)) were indicated in the figure legends.

[0048] 2. Experimental procedures and results 2.1 11-HETE monotherapy inhibits bladder tumor and enhances CD8+ T cell anti-tumor activity Figure 1 MB49 cells were subcutaneously inoculated into C57BL / 6 mice (n=6 per group), and tumor-bearing mice were intraperitoneally injected with (±) 11-HETE (500 ng per mouse) daily from day 0. The upper left is the tumor volume-time curve. The results showed that compared with the DMSO control group, the tumor growth in the (±) 11-HETE treatment group was significantly slowed down (p=0.0002). Figure 1 The upper right is the flow cytometry analysis results of tumor tissues. (±) 11-HETE treatment can significantly increase the proportion of CD3 + CD8 + / CD45 + cells in the tumor microenvironment (p=0.002), and significantly increase the proportion of IFN-γ + producing CD8 +p=0.003) with TNF-a + p=0.004), suggesting that (±) 11-HETE not only inhibited tumor progression, but also promoted CD8 + T cell infiltration and enhanced killing effect.

[0049] Figure 1 MB49 cells were subcutaneously inoculated into C57BL / 6 mice (n=6 per group). Tumor-bearing mice were intraperitoneally injected with 11(S)-HETE or 11(R)-HETE daily from day 0. The lower left side is the tumor volume curve after treatment with different chiral configurations of 11-HETE, and the results show that both 11(S)-HETE and 11(R)-HETE can significantly inhibit the growth of C57BL / 6-MB49 tumors (p<0.0001). Figure 1 The lower right side is the results of tumor tissue immune cell analysis, and both chiral configurations can increase the proportion of CD3 + CD8 + / CD45 + cells, among which 11(S)-HETE increased significantly compared with the control group (p=0.02), 11(R)-HETE increased more significantly (p=0.008), and the difference between 11(S)-HETE and 11(R)-HETE was not significant (p=0.84). In terms of functional indicators, both 11(S)-HETE and 11(R)-HETE can increase the proportion of CD8 + T cells IFN-γ + and TNF-a + ; compared with the control group, 11(S)-HETE was p=0.002 (IFN-γ + ) and p=0.009 (TNF-a + ) respectively, and 11(R)-HETE was p<0.0001 (IFN-γ + ) and p<0.0001 (TNF-a + ) respectively. Further comparison of the two chiral configurations showed that 11(R)-HETE had a stronger trend in increasing IFN-γ + (p=0.05) and TNF-a + (p=0.01), suggesting that the immune enhancement and anti-tumor activity of 11-HETE may have a chiral difference, and the effect of 11(R)-HETE is more prominent.

[0050] 2.2 11-HETE sensitizes CAR-T and inhibits bladder tumors in combination with CAR-T CAR-T cells were pre-treated with (±)11-HETE (40 ng / mL) before co-cultured with bladder cancer (BC) cells, and the level of LDH in the culture supernatant was detected by ELISA. In the same co-culture system, flow cytometry was used to analyze CAR-T induced apoptosis. In vivo experiments, SYBC1 or UMUC-3 cells were subcutaneously inoculated into NCG mice (n=6 in each group). SYBC1 tumor-bearing mice were intraperitoneally injected with (±)11-HETE (500 ng / mouse) daily from day 0, and CAR-T was adoptively transferred every 4 days from day 6, a total of 3 times, and the tumor growth curve was continuously monitored; UMUC-3 tumor-bearing mice were also intraperitoneally injected with (±)11-HETE (500 ng / mouse) daily from day 0, and CAR-T was adoptively transferred every 6 days from day 9, a total of 3 times, and the tumor growth curve was continuously monitored.

[0051] Figure 2 The upper part is the result of in vitro co-culture, CAR-CD8 + After co-culturing T cells with bladder cancer cells (SYBC1 or UMUC-3), the addition of (±)11-HETE can significantly increase the release level of LDH in the culture supernatant (indicating an increase in cell lysis / death; SYBC1: p=0.0001 compared with DMSO; UMUC-3: p<0.0001), and flow cytometry detection shows that (±)11-HETE can significantly increase the proportion of tumor cell death, and this phenomenon can be observed in SYBC1, UMUC-3 and T24 cells (SYBC1: p=0.004; UMUC-3: p=0.01; T24: p=0.02), indicating that (±)11-HETE can enhance CAR-T mediated killing of target cells. The lower part is the result of xenotransplantation model in vivo, NCG mice were inoculated with SYBC1 (5x10 5 ) or UMUC-3 (1x10 6 ) and treated with DMSO, (±)11-HETE alone, CAR-T alone and (±)11-HETE+CAR-T combination therapy (the arrow in the figure indicates the administration / transfusion time point), the tumor growth curve shows that (±)11-HETE alone has no significant antitumor effect compared with DMSO (SYBC1: p=0.61; UMUC-3: p=0.10), while the tumor volume in the combination therapy group is significantly lower than that in the CAR-T alone group, and in both models, it shows a stronger antitumor effect (SYBC1: p<0.0001; UMUC-3: p=0.005), suggesting that (±)11-HETE can significantly enhance the ability of CAR-T therapy to control the growth of bladder tumors in vivo.

[0052] 2.3 11-HETE improves bladder tumor chemotherapy response and prolongs survival Figure 3 Left in situ MB49 bladder luc Schematic diagram of experimental procedure of (±)11-HETE combined with gemcitabine plus cisplatin (GP) chemotherapy in the model. Figure 3 Left middle, right upper pair of in situ MB49 bladder luc After the model was treated with (±)11-HETE, GP chemotherapy or the combination of the two respectively, the in vivo bioluminescence imaging results were monitored, and the relative luciferase signal fold change (13th day compared with 1st day) was calculated (n=8 in each group). Figure 3 Right bottom, under the same treatment conditions, the in situ MB49 bladder luc Kaplan-Meier survival curve of the model (n=8 in each group). The results showed that, Figure 3 Middle is the representative figure of in vivo bioluminescence imaging (BLI) on the 13th day after treatment, and the four rows correspond to the four groups respectively. It can be seen from the results that the luminescence signal in the bladder region of the DMSO group is generally stronger, indicating a higher tumor burden, and individual death occurred on the 11th day; in contrast, the GP-chemo group and the (±)11-HETE group showed varying degrees of signal reduction; the overall luminescence signal of the combination treatment group was the weakest, indicating the most obvious control of the in situ tumor burden. Figure 3 Right upper is the quantitative result of BLI (fold change of luciferase signal on the 13th day compared with the 1st day, each column represents one mouse, n=8 / group), the overall trend is consistent with the imaging: the fold change of the DMSO group is the most obvious, while the fold change of the combination group is the lowest, indicating that (±)11-HETE can further enhance the tumor inhibition effect of GP-chemo. Figure 3 Right bottom is the Kaplan-Meier survival curve (n=8 / group), the survival time of the combination treatment group is the longest, better than the single drug group and the control group, indicating that (±)11-HETE combined with GP-chemo not only reduces the in situ tumor burden, but also translates into definite survival benefit in vivo.

[0053] 2.4 11-HETE combined with immunotherapy suppresses bladder tumor and activates CD8 + T cells MB49 cells were subcutaneously inoculated in C57BL / 6 mice (n=6 in each group). The tumor-bearing mice were intraperitoneally injected with (±)11-HETE (500 ng per mouse) daily from the 0th day, and anti-PD-1 antibody was injected every 3 days from the 6th day, a total of 5 times; the tumor growth curve was continuously monitored. The mice were sacrificed after the last treatment, and the tumors were removed and weighed, and flow cytometry analysis was performed on the tumor tissue to detect the proportion of CD3 + CD8 + T cells, IFNγ + / CD3 + CD8 + T cells and TNFα+ CD3 + CD8 + T cell ratio. Left panel shows that both (±)11-HETE + IgG and PD-1 neutralizing antibody can slow down tumor volume growth to different extents compared with DMSO + IgG control, while the curve of the combination group is the lowest, suggesting that the combination treatment has the strongest control of tumor growth; arrows in the curve mark the dosing time points. Figure 4 In the meantime, the scatter plot of endpoint tumor weight further verifies the above trend: the tumor weight of the combination group is the lowest, and the difference is statistically significant compared with the control and single-drug groups (the corresponding p values are given in the figure), indicating that the combination treatment can significantly reduce the overall tumor burden. The right panel is the flow cytometry result of tumor tissue: the combination group has the highest CD3 + CD8 + / CD45 + T cell ratio, suggesting that the CD8 + T cell infiltration in the tumor is enhanced; at the same time, the combination group has significantly increased CD8 + T cell IFN-γ + and TNF-α + ratios (p values are given in the figure), suggesting that their effector / killing functions are further activated. Overall, this figure shows that in bladder tumors, (±)11-HETE can enhance the anti-tumor effect of PD-1 blockade treatment, and achieve stronger immune-mediated tumor control by promoting CD8 + T cell infiltration in the tumor and increasing the production of effector factors such as IFN-γ and TNF-α.

[0054] 2.5 11-HETE combined immunotherapy inhibits colon tumors and enhances CD8 + T cell infiltration MC38 cells were subcutaneously inoculated into C57BL / 6 mice (n = 6 per group). Tumor-bearing mice were intraperitoneally injected with (±)11-HETE (500 ng per mouse) daily from day 0, and anti-PD-1 antibody was injected every 3 days from day 6, a total of 5 times; the tumor growth curve was continuously monitored. After the last treatment, the tumors were removed and weighed, and flow cytometry analysis was performed on the tumor tissue to detect CD3 + CD8 + T cell ratio, IFNγ + / CD3 + CD8 + T cell ratio, and TNFα + / CD3 + CD8 +T cell ratio. Left panel shows that both (±)11-HETE + IgG and PD-1 neutralizing antibody can slow down the tumor volume growth to different extents compared with DMSO + IgG control, while the curve of the combination group is the lowest, suggesting that the combination treatment has the strongest control on tumor growth; the arrows in the curve mark the dosing time points. Figure 5 In the meantime, the endpoint tumor weight scatter plot further verifies the above trend: the tumor weight of the combination group is the lowest, and the difference is statistically significant compared with the control and single drug groups (p values are given in the figure), indicating that the combination treatment can significantly reduce the overall tumor burden. The right panel is the flow cytometry result of tumor tissue: the combination group has the highest CD3 + CD8 + / CD45 + cell ratio, suggesting that the CD8 + T cell infiltration in the tumor is enhanced; at the same time, the combination group has significantly increased CD8 + T cell IFN-γ + and TNF-α + ratios (p values are given in the figure), suggesting that their effector / killing functions are further activated. Overall, this figure shows that in colorectal tumors, (±)11-HETE can enhance the anti-tumor effect of PD-1 blocking treatment, and achieve stronger immune-mediated tumor control by promoting CD8 + T cell infiltration in the tumor and increasing the production of effector factors such as IFN-γ and TNF-α.

[0055] 2.6 11-HETE enhances the inhibitory effect of chemotherapy combined with immunotherapy on bladder tumors MB49 cells were subcutaneously inoculated into C57BL / 6 mice (n=6 per group). The tumor-bearing mice were intraperitoneally injected with (±)11-HETE (500 ng / mouse) daily from day 6, and anti-PD-1 antibody was injected every 3 days from day 6, for a total of 5 times; at the same time, GP chemotherapy was given every 6 days from day 6, where G is 1.5 mg / kg and P is 5 mg / kg, for a total of 3 times. The tumor growth curve was continuously monitored. The results show that the DMSO + IgG control group has the fastest tumor growth; GP + IgG can delay tumor growth to a certain extent; after adding PD-1 neutralizing antibody to GP (GP + PD1 neutralizing antibody), the tumor growth is further inhibited; while the GP + PD1 neutralizing antibody + (±)11-HETE combination group has the lowest curve, suggesting that the triple regimen has the strongest control on tumor growth. The p values marked in the figure show that the anti-tumor effect of the combination group is statistically significant compared with the control or the corresponding comparison group (p<0.0001 or p<0.001), supporting that (±)11-HETE can further enhance the anti-tumor effect in vivo on the basis of PD-1 blocking (combined with chemotherapy / treatment).

[0056] It is apparent that the above-mentioned embodiments are only examples for clearly illustrating the present application and are not intended to limit the embodiments of the present application. Those skilled in the art can make various modifications and variations without departing from the spirit and scope of the present application. Thus, if these modifications and variations of the present application fall within the scope of the claims of the present application and their equivalents, it is also intended to include these modifications and variations.

Claims

1. A pharmaceutical composition having an antitumor effect or an auxiliary antitumor effect, characterized by: The pharmaceutical composition comprises 11-hydroxyeicosatetraenoic acid (11-HETE) as the only active ingredient or comprises 11-HETE and other anti-tumor drugs.

2. The pharmaceutical composition of claim 1, wherein: The 11-HETE is selected from one or more of (R) 11-HETE, (S) 11-HETE or (R / S) 11-HETE mixture.

3. The pharmaceutical composition of claim 1, wherein: The 11-HETE targets CD8 + T cells, enhances CD8 + T cell infiltration, proliferation, killing function, the 11-HETE directly promotes immune surveillance and inhibits tumor growth, and has a sensitizing effect on the other anti-tumor drugs.

4. The pharmaceutical composition of claim 1, wherein: The other anti-tumor drugs include chemotherapeutic drugs or immune anti-tumor drugs.

5. The pharmaceutical composition of claim 4, wherein: The chemotherapeutic drugs include one or more of alkylating agents, antimetabolites, anti-tumor antibiotics, plant alkaloids or platinum anti-tumor drugs, and the immune anti-tumor drugs include immune checkpoint inhibitors or CAR-T cell therapy.

6. The pharmaceutical composition of claim 5, wherein: The chemotherapeutic drugs are selected from one or more of capecitabine, gemcitabine, paclitaxel, docetaxel, cyclophosphamide, cisplatin, carboplatin, oxaliplatin or 5-FU, and the immune checkpoint inhibitors are selected from one or more of PD-1 inhibitors, PD-L1 inhibitors or CTLA-4 inhibitors.

7. Use of the pharmaceutical composition of any one of claims 1 to 6 in the preparation of an anti-tumor or adjuvant anti-tumor drug.

8. Use according to claim 7, characterized in that: The drug further comprises a pharmaceutically acceptable excipient, and the drug is an oral preparation or a parenteral preparation.

9. Use according to claim 8, characterized in that: The tumor is a solid tumor.

10. Use according to claim 9, characterized in that: The tumor is selected from one or more of bladder cancer, colon cancer, prostate cancer, glioblastoma, endometrial cancer, breast cancer, colorectal cancer, ovarian cancer, melanoma, head and neck squamous cell carcinoma or non-small cell lung cancer.

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