EI24 agonist and application of EI24 agonist in preparation of preparation for inhibiting proliferation of esophageal squamous carcinoma

Abstract

The invention belongs to the technical field of esophageal squamous carcinoma inhibitors, and particularly relates to an EI24 agonist and application of the EI24 agonist in preparation of a preparation for inhibiting proliferation of esophageal squamous carcinoma. The EI24 agonist is pentagalloylglucose, FOXP1 protein or a combination of the pentagalloylglucose and the FOXP1 protein. The expression quantity of EI24 can be improved in a targeted manner, the proliferation of esophageal squamous carcinoma cells is inhibited, and the application prospect in the aspect of inhibiting esophageal squamous carcinoma is very good.

Description

Technical Field

[0001] The invention belongs to the technical field of esophageal squamous cell carcinoma preparations, and specifically relates to an EI24 agonist and an application thereof in preparing a preparation for inhibiting the proliferation of esophageal squamous cell carcinoma. Background Art

[0002] EI24 (etoposide-induced gene 24) is a DNA damage-related gene induced by etoposide and can be directly activated by the p53 gene. EI24 protein is mainly located on the endoplasmic reticulum membrane. It is an important protein that activates autophagy and participates in the formation of cell autophagy. At the same time, EI24 is also involved in biological processes such as cell apoptosis, proliferation, drug resistance, inflammation, cell metastasis, and epithelial-mesenchymal transition.

[0003] Digestive tract tumors have the characteristics of difficult early diagnosis, easy recurrence and metastasis, and poor prognosis. The development of new and more specific digestive tract tumor marker molecules and therapeutic targets is of great significance for conquering digestive tract tumors. EI24 is a newly discovered tumor-related gene marker in recent years. It is involved in regulating many tumor biological processes. The abnormal expression of E124 is closely related to the occurrence and development of digestive tract tumors such as pancreatic cancer, liver cancer, colorectal cancer, and esophageal cancer. Inhibiting tumor cell growth and sensitizing tumor cells to chemotherapy drugs by activating EI24 expression is the key to developing tumor treatment drugs. However, the existing technology focuses more on theoretical research on the expression level of EI24 in tumor patients, such as "Wei Siyu, Hong Liu, Yang Wanli, et al. Research progress of EI24 in digestive tract tumors [J]. Modern Oncology, 2024 (006): 032" and other documents disclosed in the literature. The research and development team of the present invention has also been committed to the research related to the expression level of EI24 in tumor patients, but these have not been well transformed into practical applications, and there is a lack of agonists that can effectively activate EI24. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides an EI24 agonist and its application in preparing a preparation for inhibiting the proliferation of esophageal squamous cell carcinoma. Theoretical research is transformed into practical application level, and the EI24 agonist is used to promote the expression of EI24, showing the therapeutic sensitivity of esophageal squamous cell carcinoma.

[0005] The object of the present invention is to provide an EI24 agonist, which is pentagalloylglucose, FOXP1 protein or a combination of the two.

[0006] When the EI24 agonist is composed of pentagalloylglucose and FOXP1 protein, the mass ratio of the two is 1:1-3.

[0007] Pentagalloylglucose, FOXP1 protein or a combination of the two all showed the effect of activating EI24 expression, and are effective agonists for activating EI24. They can transform theoretical research into practical application levels, promote EI24 expression through EI24 agonists, and show therapeutic sensitivity of esophageal squamous cell carcinoma.

[0008] The present invention also provides a composition comprising an EI24 agonist, wherein the composition is a combination of an EI24 agonist and an effect enhancer;

[0009] When the effect enhancer and FOXP1 protein are combined, the mass ratio of the two is 1:5-8.

[0010] When the effect enhancer is combined with pentagalloylglucose, the mass ratio of the two is 1:6-10.

[0011] When the effect enhancer is combined with pentagalloylglucose and FOXP1 protein, the mass ratio of the three is 1:3-5:3-5.

[0012] The effect enhancer is a substance that can increase the expression of EI24; the effect enhancer is fructose. After fructose is compounded with an EI24 agonist, it can enhance the effect of increasing the expression of EI24. However, studies have shown that fructose alone cannot effectively increase the expression of EI24, indicating that there is a synergistic promotion process between fructose and the EI24 agonist in the compounding effect.

[0013] Preferably, in the above composition, when the effect enhancer and FOXP1 protein are combined, the mass ratio of the two is 1:5.

[0014] Preferably, in the above composition, when the effect enhancer is combined with pentagalloylglucose, the mass ratio of the two is 1:8.

[0015] Preferably, in the above composition, when the effect enhancer, pentagalloyl glucose and FOXP1 protein are combined, the mass ratio of the three is 1:4:4.

[0016] The present invention also provides an application of an EI24 agonist or a composition in preparing a preparation for inhibiting the proliferation of esophageal squamous cell carcinoma.

[0017] Preferably, in the above application, the inhibition of esophageal squamous cell carcinoma proliferation refers to the inhibition of proliferation of human esophageal cancer cells EC109 or human esophageal cancer cells EC9706.

[0018] Preferably, in the above application, the preparation is obtained by compounding the EI24 agonist with a pharmaceutically acceptable excipient; or, the preparation is obtained by compounding the composition with a pharmaceutically acceptable excipient.

[0019] Preferably, in the above application, the auxiliary materials include one or more of fillers, disintegrants, binders, lubricants, cosolvents, stabilizers and surfactants.

[0020] For example, the filler is one or more of lactose, sucrose and starch.

[0021] The disintegrant is one or more of sodium carboxymethylcellulose or cross-linked polyvinylpyrrolidone.

[0022] The adhesive is gelatin or polyvinyl pyrrolidone.

[0023] The lubricant is one or more of magnesium stearate, talcum powder, and micro-powder silica gel.

[0024] The cosolvent is polyethylene glycol or propylene glycol.

[0025] The stabilizer is an antioxidant or a preservative to prevent the drug from being denatured or contaminated. For example, the antioxidant is vitamin C or vitamin E, and the preservative is sodium benzoate or potassium sorbate.

[0026] The surfactant is Tween 80 or polysorbate 80.

[0027] The above are common pharmaceutically acceptable excipients. Without departing from the inventive concept of the present invention, any replacement of the above excipients made by those skilled in the art is deemed to be an equivalent replacement.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention provides an EI24 agonist, which is pentagalloylglucose, FOXP1 protein or a combination of the two. These substances or a combination of substances can target and increase the expression of EI24 and inhibit the proliferation of esophageal squamous cell carcinoma. After further combination with fructose, the effect of stimulating the expression of EI24 is more obvious, and the effect of inhibiting the proliferation of esophageal squamous cell carcinoma is also more obvious, and it has a good application prospect in inhibiting the proliferation of esophageal squamous cell carcinoma. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The results are the clinical significance analysis of EI24 expression in esophageal squamous cell carcinoma.

[0031] A, EI24 immunohistochemical staining scores of adjacent tissues and cancerous tissues of esophageal squamous cell carcinoma; B, Kaplan-Meier survival curve based on EI24 expression levels of patients with esophageal squamous cell carcinoma, log-rank test: p=0.0012.

[0032] Figure 2 These are the experimental results of ferroptosis in human esophageal cancer cells EC109.

[0033] Figure 3 These are the experimental results of ferroptosis in human esophageal cancer cells EC9706.

[0034] Figure 4 This is the WB (Western Blot) test result of PGG treatment.

[0035] Figure 5 These are the results of a WB test after treatment of the esophageal squamous cell carcinoma mouse model EC109-EO.

[0036] Figure 6 These are the results of a WB test after treating the esophageal squamous cell carcinoma mouse model EC109-NC. DETAILED DESCRIPTION

[0037] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below in conjunction with specific embodiments and drawings.

[0038] In the description of the present invention, unless otherwise specified, all reagents used are commercially available and all methods used are conventional techniques in the art.

[0039] In the description of the present invention, pentagalloyl glucose was purchased from Chengdu Caoyuankang Biotechnology Co., Ltd. FOXP1 protein was a recombinant FOXP1 protein purchased from Hubei Aipidi Bioengineering Co., Ltd., and the product name was FOXP1.

[0040] The present invention provides an EI24 agonist, which is pentagalloylglucose (PGG), FOXP1 protein (forkhead box protein P1) or a combination of the two.

[0041] These substances or combinations of substances can target and increase the expression of EI24 and inhibit the proliferation of esophageal squamous cell carcinoma.

[0042] Based on the same inventive concept, the present invention provides a composition comprising an EI24 agonist, wherein the EI24 agonist is combined with an effect enhancer to obtain an agonist composition; the effect enhancer is fructose. When the effect enhancer is combined with FOXP1 protein, the mass ratio of the two is 1:5-8; when the effect enhancer is combined with pentagalloyl glucose, the mass ratio of the two is 1:6-10; when the effect enhancer is combined with pentagalloyl glucose and FOXP1 protein, the mass ratio of the two is 1:3-5:3-5.

[0043] Based on the same inventive concept, the present invention provides an application of an EI24 agonist in the preparation of a preparation for inhibiting the proliferation of esophageal squamous cell carcinoma.

[0044] The present invention provides an application of a composition containing an EI24 agonist in preparing a preparation for inhibiting proliferation of esophageal squamous cell carcinoma.

[0045] The present invention includes the following embodiments.

[0046] Example 1

[0047] An EI24 agonist is pentagalloylglucose.

[0048] Example 2

[0049] An EI24 agonist is the FOXP1 protein.

[0050] Example 3

[0051] An EI24 agonist is provided, wherein the mass ratio of pentagalloylglucose and FOXP1 protein is 1:1. The materials are mixed to obtain the EI24 agonist.

[0052] Example 4

[0053] An EI24 agonist is provided, wherein the mass ratio of pentagalloylglucose and FOXP1 protein is 1:3. The materials are mixed to obtain the EI24 agonist.

[0054] Example 5

[0055] A composition is a combination of fructose and FOXP1 protein, wherein the mass ratio of fructose to FOXP1 protein is 1:5. The materials are combined to obtain the composition of this embodiment.

[0056] Example 6

[0057] A composition is a combination of fructose and FOXP1 protein, wherein the mass ratio of fructose to FOXP1 protein is 1:8. The materials are combined to obtain the composition of this embodiment.

[0058] Example 7

[0059] A composition is composed of fructose and pentagalloylglucose, wherein the mass ratio of fructose to pentagalloylglucose is 1:8. The materials are combined to obtain the composition of this embodiment.

[0060] Example 8

[0061] A composition is composed of fructose and pentagalloylglucose, wherein the mass ratio of fructose to pentagalloylglucose is 1:6. The materials are combined to obtain the composition of this embodiment.

[0062] Example 9

[0063] A composition is composed of fructose and pentagalloylglucose, wherein the mass ratio of fructose to pentagalloylglucose is 1:10. The materials are combined to obtain the composition of this embodiment.

[0064] Example 10

[0065] A composition is composed of fructose, pentagalloylglucose and FOXP1 protein, wherein the mass ratio of fructose, pentagalloylglucose and FOXP1 protein is 1:4:4. The materials are combined to obtain the composition of this embodiment.

[0066] Embodiment 11

[0067] A composition is composed of fructose, pentagalloylglucose and FOXP1 protein, wherein the mass ratio of fructose, pentagalloylglucose and FOXP1 protein is 1:3:3. The materials are combined to obtain the composition of this embodiment.

[0068] Example 12

[0069] A composition is composed of fructose, pentagalloylglucose and FOXP1 protein, wherein the mass ratio of fructose, pentagalloylglucose and FOXP1 protein is 1:5:5. The materials are combined to obtain the composition of this embodiment.

[0070] The effects of the present invention are described below in conjunction with some experiments.

[0071] Experiment 1: EI24 is lowly expressed in esophageal squamous cell carcinoma, and low expression of EI24 is associated with poor prognosis of esophageal squamous cell carcinoma

[0072] According to the research results of "Duan L, Ma J, Yang W, et al. EI24 Inhibits Cell Proliferation and Drug Resistance of Esophageal Squamous Cell Carcinoma. [J]. Frontiers inoncology, 2020, 10: 1570. DOI: 10.3389 / fonc.2020.01570", EI24 is lowly expressed in esophageal squamous cell carcinoma, and the survival rate of esophageal squamous cell carcinoma patients with low expression of EI24 is lower.

[0073] The present invention has been studied again with different samples, and the results are shown in Figure 1 ,Combining all the research results, it shows that EI24 is lowly expressed in esophageal squamous cell carcinoma, and low expression of EI24 is associated with poor prognosis of esophageal squamous cell carcinoma.

[0074] Experiment 2: EI24 promotes ferroptosis in esophageal squamous cell carcinoma

[0075] Test cells: human esophageal cancer cell line EC109, human esophageal cancer cell line EC9706.

[0076] Experimental method: When the above cells are cultured to a confluency of 85%, ferroptosis is induced by a ferroptosis inducer, and drug treatment is performed at the same time for 5 consecutive days, and the OD450nm value of the cell fluid is counted every day. The drug treatment settings are as follows:

[0077] EC109-LV-NC-EI24+DFO: every 10 6 Add 100 μL of 10 μg / L EI24 protein solution and 50 μL of 10 μg / L deferoxamine (DFO) to each EC109 cell.

[0078] EC109-LV-NC: EC109 cells without drug treatment.

[0079] EC9706-LV-NC-EI24+DFO: every 10 6 Add 100 μL of 10 μg / L EI24 protein solution and 50 μL of 10 μg / L DFO to each EC9706 cell.

[0080] EC9706-LV-NC: EC9706 cells without drug treatment.

[0081] Figure 2 EC109-LV-NC indicates that the cells were not treated with EI24 protein and DFO, EC109-LV-NC-EI24 indicates that the cells were treated with EI24 protein, and EC109-LV-NC-EI24+DFO indicates that the cells were treated with EI24 protein and DFO. Figure 2 The results showed that after treatment with EI24 protein, the OD450 of EC109 cells was significantly reduced. DFO is an iron chelator used to inhibit ferroptosis of EC109 cells, and can also inhibit EC109 cell death caused by EI24 protein.

[0082] Figure 3 EC9706-LV-NC means not treated with EI24 protein and DFO, EC9706-LV-NC-EI24 means treated with EI24 protein, and EC9706-LV-NC-EI24+DFO means treated with EI24 protein and DFO. Figure 3 The results showed that after treatment with EI24 protein, the OD450 of EC9706 cells was significantly reduced. DFO was used to inhibit ferroptosis of EC9706 cells and could also inhibit the death of EC9706 cells caused by EI24 protein.

[0083] Experiment 3: Experiment to increase EI24 expression

[0084] The EC109 human esophageal cancer cell line was transplanted subcutaneously into immunodeficient mice to establish the EC109-EO esophageal squamous cell carcinoma mouse model. In addition, the EC9706 human esophageal cancer cell line was transplanted subcutaneously into immunodeficient mice to establish the EC109-NC esophageal squamous cell carcinoma mouse model.

[0085] The esophageal squamous cell carcinoma mouse model EC109-EO or esophageal squamous cell carcinoma mouse model EC109-NC were treated with different concentrations of PGG, and the PGG dosage was 100 μL / kg mouse. β-ACTIN was used as an internal reference, and WB test was performed to test the expression of EI24. The results are shown in Figure 4 The results showed that PGG treatment could increase the expression of EI24, and the expression of EI24 increased with the increase of PGG concentration.

[0086] The esophageal squamous cell carcinoma mouse model EC109-EO or esophageal squamous cell carcinoma mouse model EC109-NC was treated with FOXP1 protein. The amount of FOXP1 protein solution was 100 μL / kg mouse, and the concentration of FOXP1 protein solution was 10 μg / L. β-ACTIN was used as an internal reference, and WB test was performed to test the expression of EI24. The results are shown in Figure 5 The results showed that FOXP1 protein treatment could increase the expression of EI24.

[0087] In addition, the esophageal squamous cell carcinoma mouse model EC109-EO was treated with the EI24 agonist of Example 3 and the composition of Example 5, Example 7, and Example 10; or the esophageal squamous cell carcinoma mouse model EC109-NC was treated with the EI24 agonist of Example 3 and the composition of Example 5, Example 7, and Example 10, and the dosage of the EI24 agonist or the composition was 100 μL / kg mouse, wherein the concentration of the EI24 agonist or the composition was 10 μg / L. Using β-ACTIN as an internal reference, a WB test was performed to test the expression of EI24. Results are shown in Figure 6 The results showed that whether it was the combination of PGG and FOXP1 protein or the addition of fructose, the expression of EI24 increased significantly compared with the control group, and the bands were obvious.

[0088] Experiment 4: In vitro inhibition of esophageal squamous cell carcinoma cells

[0089] Test cells: human esophageal cancer cell line EC109, human esophageal cancer cell line EC9706, and human esophageal normal cell line HET-1A.

[0090] When the cells were cultured to a confluence of 85%, the cells were treated with drugs, and the cell growth inhibition rate was calculated after 5 days of incubation compared with that before drug treatment. The drug addition settings were as follows:

[0091] Blank control: every 10 6Add 100 μL of 0.9% saline to each EC109 cell.

[0092] Positive control: every 10 6 Add 100 μL of 10 μg / L DMSO to each EC109 cell.

[0093] Example 1 Group: Every 10 6 100 μL of the EI24 agonist of Example 1 at a concentration of 10 μg / L was added to each EC109 cell.

[0094] Example 2 Group: Every 10 6 100 μL of the EI24 agonist of Example 2 at a concentration of 10 μg / L was added to each EC109 cell.

[0095] Example 3 Group: Every 10 6 100 μL of the EI24 agonist of Example 3 at a concentration of 10 μg / L was added to each EC109 cell.

[0096] Example 4 Group: Every 10 6 100 μL of the EI24 agonist of Example 4 at a concentration of 10 μg / L was added to each EC109 cell.

[0097] Example 5 Group: Every 10 6 100 μL of the composition of Example 5 at a concentration of 10 μg / L was added to each EC109 cell.

[0098] Example 6 Group: Every 10 6 100 μL of the composition of Example 6 at a concentration of 10 μg / L was added to each EC109 cell.

[0099] Example 7 Group: Every 10 6 100 μL of the composition of Example 7 at a concentration of 10 μg / L was added to each EC109 cell.

[0100] Example 8 Group: Every 10 6 100 μL of the composition of Example 8 at a concentration of 10 μg / L was added to each EC109 cell.

[0101] Example 9 Group: Every 10 6 100 μL of the composition of Example 9 at a concentration of 10 μg / L was added to each EC109 cell.

[0102] Example 10 Group: Every 10 6 100 μL of the composition of Example 10 at a concentration of 10 μg / L was added to each EC109 cell.

[0103] Example 11 Group: Every 10 6100 μL of the composition of Example 11 at a concentration of 10 μg / L was added to each EC109 cell.

[0104] Example 12 Group: Every 10 6 100 μL of the composition of Example 12 at a concentration of 10 μg / L was added to each EC109 cell.

[0105] Experimental method: The cell growth inhibition rate was determined by MTT assay.

[0106] Result processing: Each test was set up with 3 parallel treatments, and the growth inhibition rate results were averaged.

[0107] Table 1 Cell growth inhibition rate results 1

[0108]

[0109] The results in Table 1 show that Example 1 to Example 12 all have good inhibitory effects on human esophageal cancer cells EC109 and EC9706, and the inhibition rate on human esophageal normal cells HET-1A is low, indicating that the damage to normal human esophageal normal cells HET-1A is very small.

[0110] Experiment 5: In vitro ferroptosis of esophageal cancer cells

[0111] Test cells: human esophageal cancer cell line EC109, human esophageal cancer cell line EC9706.

[0112] When the cells were cultured to a confluence of 85%, the cells were treated with drugs, and the cell growth inhibition rate was calculated after 5 days of incubation compared with that before drug treatment. The drug addition settings were as follows:

[0113] Blank control: every 10 6 Add 100 μL of 0.9% saline and 50 μL of 10 μg / L DFO to each EC109 cell.

[0114] Positive control: every 10 6 Add 100 μL of 10 μg / L DMSO and 50 μL of 10 μg / L DFO to each EC109 cell.

[0115] Example 1 Group: Every 10 6 100 μL of 10 μg / L EI24 agonist of Example 1 and 50 μL of 10 μg / L DFO were added to each EC109 cell.

[0116] Example 2 Group: Every 10 6100 μL of 10 μg / L EI24 agonist of Example 2 and 50 μL of 10 μg / L DFO were added to each EC109 cell.

[0117] Example 3 Group: Every 10 6 100 μL of 10 μg / L EI24 agonist of Example 3 and 50 μL of 10 μg / L DFO were added to each EC109 cell.

[0118] Example 4 Group: Every 10 6 100 μL of 10 μg / L EI24 agonist of Example 4 and 50 μL of 10 μg / L DFO were added to each EC109 cell.

[0119] Example 5 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 5 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0120] Example 6 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 6 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0121] Example 7 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 7 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0122] Example 8 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 8 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0123] Example 9 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 9 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0124] Example 10 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 10 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0125] Example 11 Group: Every 10 6 To each EC109 cell, 100 μL of the composition of Example 11 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0126] Example 12 Group: Every 106 To each EC109 cell, 100 μL of the composition of Example 12 at a concentration of 10 μg / L and 50 μL of 10 μg / L DFO were added.

[0127] Experimental method: The cell growth inhibition rate was determined by MTT assay.

[0128] Results processing: Each test was set up with 3 parallel treatments, and the growth inhibition rate results were averaged. The results in Table 2 show that in the in vitro ferroptosis test of esophageal cancer cells, Example 1 to Example 12 groups all had good inhibitory effects on human esophageal cancer cells EC109 and human esophageal cancer cells EC9706, with high cell death rates.

[0129] Table 2 Cell growth inhibition rate results 2

[0130]

[0131]

[0132] It should be noted that when the present invention involves a numerical range, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes a preferred embodiment. Although the preferred embodiments of the present invention have been described, once those skilled in the art know the basic inventive concept, they can make other changes and modifications to these embodiments, and these changes and modifications all fall within the scope of the present invention.

[0133] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. If these modifications and variations of the present invention fall within the scope of the equivalent technology of the present invention, the present invention is also intended to include these modifications and variations.

Claims

1. An EI24 agonist, characterized in that is pentagalloylglucose, FOXP1 protein, or a combination of the two; When the EI24 agonist is composed of pentagalloylglucose and FOXP1 protein, the mass ratio of the two is 1:1-3.

2. A composition comprising the EI24 agonist according to claim 1, characterized in that The composition is a combination of an EI24 agonist and an effect enhancer; When the effect enhancer and the FOXP1 protein are combined, the mass ratio of the two is 1:5-8; When the effect enhancer is combined with the pentagalloylglucose, the mass ratio of the two is 1:6-10; When the effect enhancer, the pentagalloylglucose and the FOXP1 protein are combined, the mass ratio of the three is 1:3-5:3-5; The effect enhancer is a substance that can increase the expression of EI24; The effect enhancer is fructose.

3. The composition according to claim 2, characterized in that When the effect enhancer and the FOXP1 protein are combined, the mass ratio of the two is 1:

5.

4. The composition according to claim 2, characterized in that When the effect enhancer is combined with the pentagalloylglucose, the mass ratio of the two is 1:

8.

5. The composition according to claim 2, characterized in that When the effect enhancer, the pentagalloylglucose and the FOXP1 protein are combined, the mass ratio of the three is 1:4:

4.

6. Use of the EI24 agonist according to claim 1 or the composition according to claim 2 in the preparation of a preparation for inhibiting the proliferation of esophageal squamous cell carcinoma.

7. The use according to claim 6, characterized in that: The inhibition of esophageal squamous cell carcinoma proliferation refers to the inhibition of proliferation of human esophageal cancer cells EC109 or human esophageal cancer cells EC9706.

8. The use according to claim 6, characterized in that: The preparation is obtained by compounding the EI24 agonist with pharmaceutically acceptable excipients; Alternatively, the preparation is obtained by compounding the composition with pharmaceutically acceptable excipients.

9. The use according to claim 8, characterized in that: The auxiliary materials include one or more of fillers, disintegrants, binders, lubricants, cosolvents, stabilizers and surfactants.

10. The use according to claim 9, characterized in that: The filler is one or more of lactose, sucrose and starch; The disintegrant is one or more of sodium carboxymethylcellulose or cross-linked polyvinylpyrrolidone; The adhesive is gelatin or polyvinyl pyrrolidone; The lubricant is one or more of magnesium stearate, talcum powder, and micro-powdered silica gel; The cosolvent is polyethylene glycol or propylene glycol; The stabilizer is an antioxidant or a preservative; The surfactant is Tween 80 or polysorbate 80.

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