WO2022105763A1

WO2022105763A1 - Method for treating gastric cancer targeting cd24

Info

Publication number: WO2022105763A1
Application number: PCT/CN2021/131010
Authority: WO
Inventors: 马斌; 高维强; 冀露
Original assignee: 上海交通大学
Priority date: 2020-11-19
Filing date: 2021-11-16
Publication date: 2022-05-27
Also published as: CN114533872A; CN114533872B

Abstract

Provided in the present invention is a method for treating gastric cancer targeting CD24. Specifically provided is the use of a CD24 inhibitor for preparing a pharmaceutical composition, the pharmaceutical composition being used for the prevention and/or treatment of gastric cancer. The present invention has discovered for the first time that CD24 inhibitors can be used for the treatment of gastric cancer, thereby providing a new targeted treatment method for gastric cancer.

Description

Gastric cancer therapy targeting CD24

technical field

The invention relates to the field of medicine, in particular to a method for treating gastric cancer targeting CD24.

Background technique

Gastric cancer is the fifth most common cancer type in the world and the third most common cause of cancer-related death, with very high morbidity and mortality in East Asian countries such as China. The incidence and mortality of gastric cancer are closely related to regional and environmental factors, such as poor diet, living habits, and Helicobacter pylori infection.

Treatment options for early gastric cancer include endoscopic resection, surgery (gastrectomy), antibiotic therapy to eradicate Helicobacter pylori, and adjuvant therapy. In the past 20 years, some progress has been made in the surgical treatment, postoperative care, and multidisciplinary treatment of gastric cancer, but the prognosis of gastric cancer has improved only slightly. It is very difficult to treat advanced gastric cancer, which is the main reason for the high mortality of gastric cancer. Currently, the first-line treatment for advanced gastric cancer is chemotherapy based on platinum drugs and 5-fluorouracil (5-Fu). In addition, trastuzumab has been approved for first-line treatment of patients with human epidermal growth factor receptor 2 (HER-2)-positive gastric cancer. The drug ramucirumab, which targets vascular endothelial growth factor (VEGF), has also been approved for patients with advanced gastric cancer who have failed first-line therapy. Although there are many treatments for gastric cancer, the overall survival rate for gastric cancer worldwide is only about 20%.

In recent years, research on tumor immunotherapy has brought new hope and prospects for the treatment of cancer. However, due to the complexity of the immune microenvironment of gastric cancer and the heterogeneity of tumors, the research on immunotherapy in the treatment of gastric cancer is still very lacking.

Therefore, there is an urgent need in the art to provide new gastric cancer targeted therapy methods.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide a new gastric cancer targeted therapy method.

The first aspect of the present invention provides the use of a CD24 inhibitor for preparing a pharmaceutical composition for preventing and/or treating gastric cancer.

In another preferred embodiment, the pharmaceutical composition is used to upregulate the infiltration of cells selected from the group consisting of CD4 ⁺ T cells, CD8 ⁺ T cells, B cells, M1 macrophages, or a combination thereof.

In another preferred embodiment, the pharmaceutical composition is used to downregulate the infiltration of T (Treg) cells and/or M2 macrophages into tumor tissue.

In another preferred example, the gastric cancer is gastric cancer that is insensitive to PD-1 and/or PD-L1 inhibitors or gastric cancer that is resistant to PD-1 and/or PD-L1 inhibitors.

In another preferred embodiment, the gastric cancer is a gastric cancer with high CD24 expression.

In another preferred embodiment, the gastric cancer is selected from the group consisting of in situ gastric cancer, intestinal-type gastric cancer, diffuse gastric cancer, gastric adenocarcinoma, Helicobacter pylori-induced gastric cancer, or a combination thereof.

In another preferred example, the gastric cancer is early gastric cancer, intermediate gastric cancer or advanced gastric cancer.

In another preferred embodiment, the inhibitor is selected from the group consisting of: an antibody or small molecule inhibitor targeting CD24 and/or its receptor protein; a targeting nucleic acid molecule targeting CD24 and/or its receptor gene or a gene editor; or a combination thereof.

In another preferred embodiment, the small molecule inhibitor includes small molecule inhibitor compounds and pharmaceutically acceptable salts thereof.

In another preferred embodiment, the CD24 receptor is sialic acid-binding immunoglobulin-like lectin 10 (Siglec-10).

In another preferred embodiment, the antibody is selected from the group consisting of polyclonal antibodies, monoclonal antibodies, chimeric antibodies, bispecific antibodies, antibody conjugates, small molecule antibodies, antibody fusion proteins, and combinations thereof.

In another preferred embodiment, the small molecule antibody is selected from the group consisting of single-chain antibody ScFv, Fab antibody, Fv fragment, and combinations thereof.

In another preferred embodiment, the ScFv antibody includes a secretory single-chain antibody expressed (including overexpressed) in the treatment cells.

In another preferred example, the therapeutic cells include mesenchymal stem cells, CAR-T cells, TCR-T cells, (CAR-)NK cells, macrophages, and dendritic cells.

In another preferred embodiment, the inhibitor is selected from the group consisting of: plant extract inhibitor, small molecule compound inhibitor, nucleic acid inhibitor, peptide inhibitor, polysaccharide inhibitor, viral vector inhibitor, lipid inhibitor Plastid carrier inhibitor, or nanoparticle carrier inhibitor.

In another preferred embodiment, the vector includes: bacterial plasmid, bacteriophage, yeast plasmid, plant cell virus, mammalian cell virus such as adenovirus, retrovirus, or other vectors.

In another preferred embodiment, the pharmaceutical composition further includes a second therapeutic agent, wherein the second therapeutic agent is selected from the group consisting of PARP1/2 inhibitors, chemotherapeutic drugs that induce DNA damage in cancer cells, DNA alkyl groups Chemotherapy drugs, DNA or RNA synthesis inhibitors, EGFR, ALK or FGFR tyrosine receptor kinase inhibitors, KRAS, MEK or ERK tumor signaling pathway inhibitors, tumor immunotherapy drugs (such as PD-1 inhibitors, PD -L ₁ inhibitor, etc.).

In another preferred embodiment, the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from the following group: liquid preparations (such as solution, emulsion, suspension), solid preparations (such as freeze-dried preparations).

In another preferred embodiment, the dosage form of the pharmaceutical composition is selected from the following group: injection, powder injection, capsule, tablet, pill, powder, granule, syrup, oral liquid or tincture.

A second aspect of the present invention provides a method for treating gastric cancer, comprising the steps of: administering a therapeutically effective amount of a CD24 inhibitor to a subject in need, thereby treating gastric cancer.

In another preferred example, the subject is a mammal.

In another preferred embodiment, the subject is selected from the group consisting of human, rat or mouse.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (eg, the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, it is not repeated here.

Description of drawings

Figure 1. (A) The mRNA expression level of CD24 in gastric adenocarcinoma was higher than that in healthy controls in TCGA database analysis; (B-C) in mouse models induced by Helicobacter Helicobacter felis (H. felis) and carcinogen MNU and in normal controls In mice, the expression of CD24 in gastric epithelial cells was analyzed by flow cytometry, and it was found that the expression of CD24 on the surface of gastric epithelial cell membrane in H.felis/MNU gastric cancer model was higher than that in normal mice. **p<0.001, ****p<0.0001.

Figure 2. (A) Analysis in the TCGA database found that the mRNA expression level of CD24 in gastric adenocarcinoma was significantly higher than that of the other two immunosuppressive molecules, CD47 and PD-L1; (B) in Helicobacter Helicobacter felis (H. felis) and oncogenic In the mouse model induced by MNU, the expression of three molecules in gastric epithelial cells was analyzed by flow cytometry, and it was found that the expression of CD24 was significantly higher than that of CD47 and PD-L1. **p<0.001, ***p<0.001.

Figure 3. In a mouse model induced by Helicobacter Helicobacter felis (H. felis) and the carcinogen MNU, tumor-forming mice were intraperitoneally injected with CD24 antibody, PD-1 antibody or IgG control, after 6 weeks of treatment It was found that CD24 antibody can significantly reduce the area of gastric tumor, while PD-1 antibody had no significant effect. *p<0.01, N.S. = not significant.

Figure 4. In a mouse model induced by Helicobacter Helicobacter felis (H. felis) and the carcinogen MNU, tumor-forming mice were intraperitoneally injected with CD24 antibody, PD-1 antibody or IgG control, after 6 weeks of treatment HE staining of gastric tissue sections showed that CD24 antibody could significantly reduce the size of gastric tumors and the abnormal phenotype of glandular structure, while PD-1 antibody had no significant effect.

Figure 5. In a mouse model induced by Helicobacter Helicobacter felis (H. felis) and carcinogen MNU, tumor-forming mice were intraperitoneally injected with CD24 antibody or IgG control, and gastric tissue was flown after 6 weeks of treatment. Cytometry analysis showed that CD24 antibody can significantly increase the proportion of CD4T, CD8T, B cells, and M1 macrophages and other immune cells with anti-tumor functions, and can reduce tumor-promoting regulatory T cells and M2 macrophages. Proportion. *p<0.01.

Detailed ways

After extensive and in-depth research and extensive screening and testing, the present inventors provide a gastric cancer treatment method by targeting CD24. The present invention finds for the first time that CD24 is significantly highly expressed in gastric cancer cells, and the administration of CD24 antibody can effectively treat gastric cancer. Further, it was found that CD24 antibody can significantly up-regulate the proportion of immune cells with anti-tumor functions such as CD4T, CD8T, B cells and M1-type macrophages, and can up-regulate the proportion of tumor-promoting regulatory T cells and M2-type macrophages. thereby inhibiting gastric cancer. In addition, CD24 inhibitors were also found to be able to suppress gastric cancers that are not sensitive to PD-1 and/or PD-L1 inhibitors. The present invention has been completed on this basis.

the term

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, when used in reference to a specifically recited value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 and (eg, 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the terms "containing" or "including (including)" can be open, semi-closed, and closed. In other words, the term also includes "consisting essentially of," or "consisting of."

As used herein, the term "room temperature" refers to a temperature of 4-40°C, preferably, 25±5°C.

As used herein, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and/or animals without undue adverse side effects (eg, toxicity, irritation, and allergy), ie, with a reasonable benefit/risk ratio.

As used herein, the term "up-regulate" refers to an increase in the amount of an indicator by 1.2-fold, preferably, 1.5-fold, 2.0-fold or 3.0-fold, compared to not administering the active ingredient of the present invention.

As used herein, the term "down-regulated" refers to a reduction in the amount of a certain indicator by a factor of 0.8, preferably, by a factor of 0.5 or 0.3, compared to not administering the active ingredient of the present invention.

CD24

CD24, also known as a thermostable antigen, is a highly glycosylated glycosylphosphatidylinositol-anchored cell surface protein. CD24 interacts with Siglec-10 molecules on innate immune cells to transmit immunosuppressive signals to suppress inflammatory responses.

CD24 inhibitor

CD24 inhibitors of the present invention include antibodies or small molecule inhibitors targeting CD24 and/or its receptor proteins; targeting nucleic acid molecules or gene editors targeting CD24 and/or its receptor genes; or combinations thereof.

Particularly preferred are CD24 monoclonal antibodies, Siglec-10 monoclonal antibodies, or a combination thereof.

Pharmaceutical composition and application

The pharmaceutical composition of the present invention contains the above-mentioned CD24 inhibitor as an active ingredient.

Experiments have proved that the pharmaceutical composition of the present invention has a significant effect on the treatment of gastric cancer, and can be used as a drug for the treatment of gastric cancer.

In particular, the pharmaceutical composition of the present invention can be used for the treatment of gastric cancer in patients who are insensitive to PD-1 and/or PD-L1 inhibitors or resistant to PD-1 and/or PD-L1 inhibitors .

The pharmaceutical composition of the present invention comprises the inhibitor of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier within a safe and effective amount.

As used herein, the term "therapeutically effective dose" refers to any amount of a drug, as described below, that, when used alone or in combination with another therapeutic agent, promotes disease regression, which manifests as disease symptoms decrease the severity of disease, increase the frequency and duration of disease-free periods, or prevent impairment or disability resulting from the disease.

A "therapeutically effective dose" of a drug of the present invention also includes a "prophylactically effective dose", which is any amount of the drug as described below, when the amount of the drug is administered alone or in combination with another therapeutic agent In a subject at risk of developing the disease or suffering from relapse of the disease, the occurrence or relapse of the disease can be inhibited.

Usually, the pharmaceutical composition contains 1-2000 mg of the active ingredient/dose of the present invention, more preferably, 10-500 mg of the active ingredient/dose of the present invention. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" means: one or more compatible solid or liquid filler or gel substances which are suitable for human use and which must be of sufficient purity and sufficiently low toxicity. "Compatibility" as used herein means that the components of the composition can be blended with the active ingredients of the present invention and with each other without significantly reducing the efficacy of the active ingredients. Examples of pharmaceutically acceptable carrier moieties include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid) , magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween)

), wetting agents (such as sodium lauryl sulfate), colorants, flavors, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The mode of administration of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration medicine.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active ingredient is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or extenders, such as , starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders such as hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants , for example, glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) ) absorption accelerators, such as quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, Calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active ingredient in such compositions may be in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be employed are polymeric substances and waxes. If desired, the active ingredient may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active active ingredient, liquid dosage forms may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-Butanediol, dimethylformamide and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures of these substances, and the like.

Besides these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

Suspensions, in addition to the active ingredient, may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like .

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the active ingredients of the present invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required if necessary.

The active ingredients of the present invention may be administered alone or in combination with other pharmaceutically acceptable therapeutic agents. In another preferred embodiment, the pharmaceutical composition further comprises one or more anticancer agents and/or immunosuppressive agents, preferably, the anticancer agents and/or immunosuppressive agents are selected from the group consisting of: PARP1/ 2 Inhibitors, chemotherapy drugs that induce DNA damage in cancer cells, DNA alkylation drugs, DNA or RNA synthesis inhibitors, EGFR, ALK or FGFR tyrosine receptor kinase inhibitors, KRAS, MEK or ERK tumor signaling pathways Inhibitors, tumor immunotherapy drugs (such as PD-1 inhibitors, PD-L ₁ inhibitors, etc.).

In another preferred embodiment, the pharmaceutical composition further comprises one or more anticancer agents and/or immunosuppressive agents, preferably, the anticancer agents and/or immunosuppressive agents are selected from the group consisting of: Panib, Lukapanib, Niraparib, Methotrexate, Capecitabine, Gemcitabine, Deoxyfluridine, Pemetrexed Disodium, Pazopanib, Imatinib, Errol tinib, lapatinib, gefitinib, vandetanib, herceptin, bevacizumab, rituximab, trastuzumab, paclitaxel, vinorelbine, docetaxel, Doxorubicin, Hydroxycamptothecin, Mitomycin, Epirubicin, Pirarubicin, Bleomycin, Letrozole, Tamoxifen, Fulvestrant, Triptorelin, Fluta Amine, leuprolide, anastrozole, ifosfamide, busulfan, cyclophosphamide, carmustine, nimustine, semustine, nitrogen mustard, melphalan, leucovorin, carboplatin , Cisplatin, Oxaliplatin, Triboplatin, Topotecan, Camptothecin, Topotecan, Everolimus, Sirolimus, Tetracarcinoma, 6-mercaptopurine, 6-thioguanine, sulfur Azathioprine, mycocin D, daunorubicin, doxorubicin, mitoxantrone, lucidomycin, prucamycin, or aminolutamide; such as nivolumab, pembrolizumab ), ipilimumab, avelumab, durvalumab, atezolizumab, or pidilizumab, or a combination thereof.

In certain embodiments, an active ingredient of the present invention is administered to a cancer-affected subject in combination with other conventional cancer treatments, eg, radiation therapy or surgery. Radiation therapy is well known in the art and includes X-ray therapy, such as gamma radiation, and radiopharmaceutical therapy.

In certain embodiments, the active ingredients of the present invention are administered concomitantly with, or sequentially with, other agents as part of a combination therapy regimen, in the same or separate formulations.

Typical ranges for therapeutically effective doses of compositions of active ingredients of the present invention will be: about 1-2000 mg/day, about 10 to about 1000 mg/day, about 10 to about 500 mg/day, about 10 to about 250 mg/day, about 10 to about 100 mg/day, or about 10 to about 50 mg/day. A therapeutically effective dose will be administered in one or more doses. It is to be understood, however, that the particular dosage of a compound of the present invention for any particular patient will depend on a variety of factors, eg, the age, sex, weight, general health, diet, individual response, time of administration, and treatment of the patient to be treated. disease severity, activity of the particular compound administered, dosage form, mode of application, and concomitant medications. A therapeutically effective amount for a given situation can be determined using routine experimentation and is within the ability and judgment of the clinician or physician. In any event, the compound or composition will be administered in multiple doses based on the individual condition of the patient and in a manner that allows delivery of a therapeutically effective amount.

The main advantages of the present invention include:

1. The present invention finds for the first time that a CD24 inhibitor can effectively treat gastric cancer, and provides a therapeutic method for treating gastric cancer by targeting CD24. In particular, the method of the present invention can be used to treat intermediate and advanced gastric cancer.

2. The present invention also finds that CD24 inhibitors can treat gastric cancer that is not sensitive to PD-1 inhibitors, thereby providing a new treatment method for such patients.

The present invention will be further described below in conjunction with specific implementation. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental method of unreceipted specific conditions in the following examples, usually according to conventional conditions, such as people such as Sambrook, molecular cloning: conditions described in laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989), or according to manufacture conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise indicated.

general approach

Flow analysis: Mouse gastric tissue was treated with tissue homogenizer gentleMACS Octo dissociator (Miltenyi Biotec) and 1 mg/mL collagenase VI (Thermo Fisher Scientific) and 50 mg/mL DNase I (Sigma-Aldrich), dispersed into single cells The suspension was subsequently treated with erythrocyte lysate (BioLegend). Fluorescence data were acquired on a FACS Aria II cytometer (BD Biosciences) and data analysis was performed using FlowJo software.

Flow antibody details are as follows:

CD3e(145-2C11), CD11c(N418), CD11b(M1/70), CD279(PD-1; 29F.1A12), F4/80(BM8), CD24(M1/69) and MHC-II(I-A/ I-E; M5/114.15.2) from BioLegend; CD4 (GK1.5) and CD8a (53-6.7) from BD Biosciences; CD45 (30-F11) and Ly-6G/Ly-6C (Gr1; RB6-8C5) ) were purchased from eBioscience.

Immunohistochemistry: The gastric tissue of fresh gastric cancer mice was fixed with 4% paraformaldehyde for 2 hours, then dehydrated with alcohol gradient, cleared with xylene, embedded in paraffin and sliced. Tissue sections were deparaffinized and rehydrated, treated with hematoxylin and eosin (Biyuntian), and photographed under a microscope (Zeiss LSM 710).

Example 1

A mouse model of Helicobacter pylori (H. felis) and N-methyl-N-nitrosourea (MNU)-induced gastric cancer: wild-type C57BL/6 mice were infected with Helicobacter felis by gavage (H. felis) (ATCC 49179). Two weeks after H. felis infection, mice were given drinking water containing 240 ppm MNU every other week for a total of 12 weeks (a total of 6 weeks of exposure), and a gastric cancer (middle and advanced stage) mouse model was obtained 36 weeks after the start of MNU treatment.

Analysis of gastric adenocarcinoma nucleic acid sequencing data in the TCGA database, as shown in Figure 1A, found that the mRNA expression of CD24 was significantly up-regulated in gastric cancer samples compared with normal gastric tissues.

In mouse models induced by Helicobacter Helicobacter felis (H. felis) and carcinogen MNU, as well as in normal control mice, the expression of CD24 in gastric epithelial cells was analyzed by flow cytometry, as shown in Figure 1B-C. The expression of CD24 protein in gastric epithelium of gastric cancer mice was significantly higher than that of normal controls. These results indicated that the expression of CD24 was significantly up-regulated during gastric carcinogenesis.

In addition, as shown in Figure 2A, the analysis in the TCGA database found that the expression level of CD24 mRNA in gastric cancer tissues was higher than that of the other two immunosuppressive molecules CD47 and PD-L1. In H.felis and MNU-induced mouse gastric cancer models (36 weeks after MNU treatment), it was also found that the expression of CD24 in E-Cadherin-positive epithelial cells was higher than that of CD47 and PD-L1 by flow cytometry analysis. This suggests that CD24 may be a more ideal therapeutic target than CD47 and PD-L1 in gastric cancer.

Example 2

Treatment regimen for gastric cancer mice: For antibody treatment, starting from 37 weeks after starting to treat mice with MNU drinking water, 50 μg CD24 monoclonal antibody/mouse/needle (R&D Systems), 100 μg PD-1 monoclonal antibody was injected twice a week Clone antibody/only/needle (Bio X Cell) or control IgG. Continuous treatment for 6 weeks. Mice were sacrificed after treatment, and gastric tissue was examined.

As shown in Figure 3, in the gastric cancer mouse model, CD24 antibody, PD-1 antibody or IgG control were intraperitoneally injected into the tumor-forming mice. The PD-1 antibody had no significant effect. This suggests that CD24 antibody can be used for the treatment of advanced gastric cancer, especially gastric cancer that is insensitive and/or resistant to PD-1.

The immunostaining results are shown in Figure 4. HE staining of gastric tissue sections showed that CD24 antibody could significantly reduce the size of gastric tumors and the abnormal phenotype of glandular structure, while PD-1 antibody had no significant effect.

The results of flow cytometry analysis are shown in Figure 5. The results show that CD24 antibody can significantly increase the proportion of immune cells with anti-tumor functions such as CD4T, CD8T, B cells and M1 macrophages, and can reduce the tumor-promoting regulatory T The proportion of cells and M2 macrophages.

In summary, the expression of CD24 in gastric epithelial cells of gastric cancer mice was significantly higher than that of control samples, and was also significantly higher than that of PD-L1 and CD47. This phenomenon was also consistent in the human tumor TCGA database. .

Through the treatment of gastric cancer mouse model induced by Helicobacter felis (H. felis) and carcinogen MNU, CD24 antibody was found to have a very good therapeutic effect. After flow cytometry analysis, it was found that the mechanism by which CD24 exerts its effect is to increase the infiltration of effector T cells (especially CD4 ⁺ and CD8 ⁺ T cells) and reduce the infiltration of regulatory T cells. In addition, the results of treatment found that treatment also It promoted the transformation of tumor-associated macrophages from M2 type to M1 type, suggesting that by CD24 antibody treatment, the immune effect was aroused in mice to fight against the immunosuppressed tumor microenvironment, inhibit tumor progression, and achieve good therapeutic effects.

discuss

The thermostable antigen CD24 is an important tumor stem cell marker in tissues and tumor stem cells, and is closely related to tumor metastasis and invasion. In addition, it is widely used as a marker for differentiating hematopoietic cells and neuronal cells. High levels of CD24 expression are found in many different tumor types, and in a new study, researchers have also identified CD24 as a new 'don't eat me' signal ), suggesting that CD24 is a potential target for cancer immunotherapy.

Although CD24 is usually widely expressed in tumor cells, it does not mean that inhibiting CD24 can definitely have a therapeutic effect on cancer. Cancers such as leukemia are not sensitive to CD24 inhibitors, so the method of inhibiting CD24 for cancer treatment still needs further research.

And the present invention proves the remarkable curative effect of CD24 inhibitor on gastric cancer (especially middle-advanced gastric cancer) through experiments for the first time, thereby providing a new gastric cancer treatment method.

All documents mentioned herein are incorporated by reference in this application as if each document were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

A use of a CD24 inhibitor, characterized in that it is used to prepare a pharmaceutical composition for preventing and/or treating gastric cancer.
The use according to claim 1, wherein the pharmaceutical composition is used to up-regulate the infiltration of cells selected from the group consisting of CD4 + T cells, CD8 + T cells, B cells, M1 type macrophages cells, or a combination thereof.
The use according to claim 1, wherein the gastric cancer is gastric cancer insensitive to PD-1 and/or PD-L1 inhibitor or resistant to PD-1 and/or PD-L1 inhibitor gastric cancer.
The use of claim 1, wherein the gastric cancer is selected from the group consisting of in situ gastric cancer, intestinal-type gastric cancer, diffuse gastric cancer, gastric adenocarcinoma, Helicobacter pylori-induced gastric cancer, or a combination thereof.
The use according to claim 1 or 3, wherein the gastric cancer is early gastric cancer, intermediate gastric cancer or advanced gastric cancer.
The use according to claim 1, wherein the inhibitor is selected from the group consisting of: antibody or small molecule inhibitor targeting CD24 and/or its receptor protein; targeting CD24 and/or its receptor gene A targeted nucleic acid molecule or gene editor; or a combination thereof.
The use of claim 1, wherein the CD24 receptor is sialic acid-binding immunoglobulin-like lectin 10 (Siglec-10).
The use of claim 1, wherein the antibody is selected from the group consisting of polyclonal antibody, monoclonal antibody, chimeric antibody, bispecific antibody, antibody conjugate, small molecule antibody, antibody fusion protein , and their combinations.
The use of claim 1, wherein the pharmaceutical composition further comprises a second therapeutic agent, wherein the second therapeutic agent is selected from the group consisting of PARP1/2 inhibitors, DNA damage-inducing agents in cancer cells Chemotherapy drugs, DNA alkylation drugs, DNA or RNA synthesis inhibitors, EGFR, ALK or FGFR tyrosine receptor kinase inhibitors, KRAS, MEK or ERK tumor signaling pathway inhibitors, tumor immunotherapy drugs (such as PD -1 inhibitors, PD-L 1 inhibitors, etc.).
The use of claim 1, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.
The use according to claim 1, wherein the dosage form of the pharmaceutical composition is selected from the group consisting of injection, powder injection, capsule, tablet, pill, powder, granule, syrup, oral liquid or tincture.
A method for treating gastric cancer, comprising the steps of: administering a therapeutically effective amount of a CD24 inhibitor to a subject in need, thereby treating gastric cancer.
The method of claim 12, wherein the gastric cancer is gastric cancer insensitive to PD-1 and/or PD-L1 inhibitor or resistant to PD-1 and/or PD-L1 inhibitor gastric cancer.