HMGB3 Inhibitor and Its Application in the Preparation of Drugs for Inhibiting the Proliferation and Migration of Esophageal Cancer Cells

Abstract

The present invention belongs to the technical field of esophageal cancer inhibitors, specifically relating to an HMGB3 inhibitor and its use in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells. The present invention discovered that HMGB3 expression is significantly increased in esophageal squamous cell carcinoma tissue compared to adjacent adjacent tissue. Therefore, inhibiting HMGB3 expression can inhibit the proliferation and migration of esophageal cancer cells. The HMGB3 inhibitor provided by the present invention can target and inhibit HMGB3 expression, thereby inhibiting the proliferation and migration of esophageal squamous cell carcinoma cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of esophageal cancer inhibitors, and particularly relates to an HMGB3 inhibitor and its application in preparing a drug for inhibiting the proliferation and migration of esophageal cancer cells. Background Art

[0002] Esophageal cancer is a general term for diseases, while esophageal squamous cell carcinoma (ESCC) is a specific histopathological classification. Clinically, most esophageal cancers are squamous cell carcinomas, or squamous cell carcinomas, with a minority being adenocarcinomas. Esophageal squamous cell carcinoma, or ESCC, refers to a malignant esophageal tumor caused by dysplasia of the esophageal squamous epithelium. Early clinical symptoms include dysphagia, foreign body sensation, and retrosternal pain. Late-stage symptoms include progressive dysphagia, with or without pain.

[0003] HMGB3 is the abbreviation of high mobility group protein B3, which is a member of the high mobility group protein (HMG). These proteins can change DNA structure and promote the binding of transcription factors, thus playing an important role in the life activities of cells. Studies have shown that HMGB3 is highly expressed in a variety of human cancers, including breast cancer, gastric cancer, non-small cell lung cancer, bladder cancer, etc. Studies have shown that the expression level of HMGB3 is abnormally and significantly increased in a variety of tumor tissues. For details, please refer to the research results of "Lv Jun. Study on the mechanism of circRNA HMGB3 regulating the proliferation, metastasis and radiotherapy resistance of esophageal cancer [D]. Guangxi Medical University, 2022", "Jianxun L, Liling W, Xuesong L. HMGB3 promotes the proliferation and metastasis of glioblastoma and is negatively regulated by miR-200b-3p and miR-200c-3p [J]. Cell biochemistry and function, 2018 (7): 36. DOI: 10.1002 / cbf.3355". However, the detailed relationship between HMGB3 and esophageal cancer is still unclear.

[0004] In addition, esophageal cancer is difficult to diagnose early, prone to recurrence and metastasis, and has a poor prognosis. Developing new agents that act on therapeutic targets for esophageal cancer is of great significance for conquering esophageal cancer. Summary of the Invention

[0005] To solve the above technical problems, the present invention provides an HMGB3 inhibitor and its use in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells. HMGB3 inhibitors act on HMGB3, a therapeutic target for esophageal cancer, and can effectively inhibit the proliferation and migration of esophageal cancer cells.

[0006] The first object of the present invention is to provide an HMGB3 inhibitor, wherein the HMGB3 inhibitor is pentagalloylglucose, an alcohol extract of Pueraria lobata flower, or a combination of the two;

[0007] When the HMGB3 inhibitor is composed of pentagalloylglucose and the alcohol extract of Pueraria lobata flower, the mass ratio of the two is 1:2-4.

[0008] Preferably, the alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0009] Fresh kudzu flower is taken, washed, dried, and crushed into 60-100 mesh powder, then mixed with 60%-70% ethanol solution, allowed to stand for 6h-12h, filtered to remove precipitation, and the collected filtrate is concentrated and freeze-dried to obtain the kudzu flower alcohol extract.

[0010] Preferably, the drying is carried out at 30° C. to 40° C. to avoid high temperature destruction of active ingredients in the kudzu flower.

[0011] Preferably, the material-liquid ratio of the powder to the ethanol solution with a volume fraction of 60% to 70% is 1 kg:8 L.

[0012] Preferably, the freeze-drying condition is to freeze-dry at -20°C.

[0013] Preferably, the kudzu flower is the unopened flower bud of the leguminous plant kudzu or the unopened flower bud of the sweet kudzu vine. The unopened flower bud is in a wrapped state, and the effective substance is less likely to diffuse into the environment.

[0014] A second objective of the present invention is to provide a method for preparing a drug for inhibiting the proliferation and migration of esophageal cancer. The present invention clearly demonstrates that HMGB3 expression is significantly increased in esophageal squamous cell carcinoma tissue compared to adjacent adjacent tissues, indicating that HMGB3 expression is highly correlated with esophageal cancer. Therefore, HMGB3 can serve as a therapeutic target for esophageal cancer, and inhibiting HMGB3 expression can inhibit the proliferation and migration of esophageal cancer cells.

[0015] Preferably, the esophageal cancer cells are one or more of human esophageal cancer cells EC109, human esophageal cancer cells EC9706, and human esophageal cancer cells ECA109.

[0016] Preferably, the drug is obtained by compounding the HMGB3 inhibitor with pharmaceutically acceptable excipients.

[0017] The auxiliary materials include one or more of fillers, disintegrants, binders, lubricants, cosolvents, stabilizers and surfactants.

[0018] For example, the filler is one or more of lactose, sucrose and starch.

[0019] The disintegrant is one or more of sodium carboxymethylcellulose or cross-linked polyvinylpyrrolidone.

[0020] The adhesive is gelatin or polyvinyl pyrrolidone.

[0021] The lubricant is one or more of magnesium stearate, talc, and micro-powdered silica gel.

[0022] The cosolvent is polyethylene glycol or propylene glycol.

[0023] 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.

[0024] The surfactant is Tween 80 or polysorbate 80.

[0025] 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 will be deemed as an equivalent replacement.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The present invention provides an HMGB3 inhibitor comprising pentagalloylglucose, an alcohol extract of Pueraria lobata flower, or a combination thereof. This HMGB3 inhibitor can target HMGB3 expression, inhibiting the proliferation and migration of esophageal cancer cells. This novel agent acts on this therapeutic target and is of great significance for combating esophageal cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The IHC score of HMGB3 immunohistochemical staining of adjacent tissues and cancerous tissues of esophageal squamous cell carcinoma was obtained.

[0029] Figure 2 Kaplan-Meier survival curves were drawn according to HMGB3 expression levels, and the log-rank test showed P < 0.01.

[0030] Figure 3 The WB (Western Blot) test results of HMGB3 expression in different cells.

[0031] Figure 4These are the results of Transwell cell migration / invasion assay 1.

[0032] Figure 5 The results of WB test show that PGG inhibits HMGB3 expression.

[0033] Figure 6 These are the WB test results showing that the alcohol extract of Pueraria lobata flower inhibits the expression of HMGB3.

[0034] Figure 7 These are the WB test results of Example 4 and Example 10 for inhibiting HMGB3 expression. DETAILED DESCRIPTION

[0035] 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 with reference to specific embodiments and drawings.

[0036] 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.

[0037] In the description of the present invention, pentagalloylglucose was purchased from Chengdu Caoyuankang Biotechnology Co., Ltd.

[0038] The present invention provides an HMGB3 inhibitor, which is pentagalloylglucose (PGG), an alcohol extract of kudzu flower, or a combination of the two.

[0039] These substances or combinations of substances can target and inhibit the expression of HMGB3, thereby inhibiting the proliferation and migration of esophageal cancer cells. Combining multiple substances can significantly inhibit HMGB3 expression and stimulate its expression.

[0040] Based on the same inventive concept, the present invention provides the use of an HMGB3 inhibitor in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells. By inhibiting the expression of HMGB3, the proliferation and migration of esophageal cancer cells, a digestive tract tumor, can be inhibited, thereby achieving the purpose of treating esophageal cancer.

[0041] The present invention includes the following embodiments.

[0042] Example 1

[0043] An HMGB3 inhibitor is pentagalloylglucose.

[0044] Example 2

[0045] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0046] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0047] Fresh kudzu flowers, i.e., open flowers of Pueraria lobata, are taken, washed to remove dust and other impurities, the pedicels and petals are retained, dried at 80°C, crushed into 60-mesh powder, then mixed with 60% by volume ethanol solution, allowed to stand for 6 hours, filtered through a 120-mesh filter to remove precipitates, the collected filtrate is concentrated, and freeze-dried at -20°C to obtain the kudzu flower alcohol extract, which is stored at -20°C for later use.

[0048] Example 3

[0049] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0050] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0051] Take fresh kudzu flowers, i.e., unopened flower buds of sweet kudzu vine, wash them to remove dust and other impurities, retain the pedicels and petals, dry them at 80°C, crush them into 60-mesh powder, then mix them with 60% ethanol solution by volume, let them stand for 6 hours, filter them with a 120-mesh filter to remove the precipitate, concentrate the collected filtrate, and freeze-dry at -20°C to obtain the kudzu flower alcohol extract, which is stored at -20°C for future use.

[0052] Example 4

[0053] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0054] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0055] Take fresh kudzu flowers, i.e., unopened flower buds of sweet kudzu vine, wash to remove dust and other impurities, retain the pedicels and petals, dry at 30°C, crush into 60-mesh powder, then mix with 60% by volume ethanol solution, let stand for 6 hours, filter with 120-mesh filter to remove precipitate, concentrate the collected filtrate, and freeze-dry at -20°C to obtain the kudzu flower alcohol extract, which is stored at -20°C for future use.

[0056] Example 5

[0057] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0058] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0059] Take fresh kudzu flowers, i.e., unopened buds of Pueraria lobata, wash them to remove dust and other impurities, retain the pedicels and petals, dry them at 40°C, grind them into 60-mesh powder, then mix them with 60% ethanol solution by volume, let them stand for 6 hours, filter them with a 120-mesh filter to remove the precipitate, concentrate the collected filtrate, and freeze-dry at -20°C to obtain the kudzu flower alcohol extract, which is stored at -20°C for future use.

[0060] Example 6

[0061] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0062] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0063] Fresh kudzu flowers, i.e., unopened flower buds of sweet kudzu vine, were taken, washed to remove dust and other impurities, the pedicels and petals were retained, dried at 30°C, crushed into 100-mesh powder, and then mixed with 60% by volume ethanol solution, allowed to stand for 6 hours, filtered through a 120-mesh filter to remove the precipitate, the collected filtrate was concentrated, and freeze-dried at -20°C to obtain the kudzu flower alcohol extract, which was stored at -20°C for later use.

[0064] Example 7

[0065] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0066] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0067] Fresh kudzu flowers, i.e., unopened buds of Pueraria lobata, were taken, washed to remove dust and other impurities, while retaining the pedicels and petals. The flowers were dried at 30°C and crushed into a 60-mesh powder. The powder was then mixed with a 70% by volume ethanol solution, allowed to stand for 6 hours, filtered through a 120-mesh filter to remove the precipitate, and the collected filtrate was concentrated and freeze-dried at -20°C to obtain the kudzu flower alcohol extract.

[0068] Example 8

[0069] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0070] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0071] Fresh kudzu flowers, i.e., unopened buds of Pueraria lobata, were taken, washed to remove dust and other impurities, while retaining the pedicels and petals. The flowers were dried at 30°C and crushed into a 60-mesh powder. The powder was then mixed with a 60% by volume ethanol solution, allowed to stand for 12 hours, filtered through a 120-mesh filter to remove the precipitate, and the collected filtrate was concentrated and freeze-dried at -20°C to obtain the kudzu flower alcohol extract.

[0072] Example 9

[0073] An HMGB3 inhibitor, an alcohol extract of Pueraria lobata flowers.

[0074] The alcohol extract of Pueraria lobata flower is prepared according to the following method:

[0075] Fresh kudzu flowers, i.e., unopened buds of Pueraria lobata, were taken, washed to remove dust and other impurities, while retaining the pedicels and petals. The flowers were dried at 30°C and crushed into a 60-mesh powder. The powder was then mixed with a 60% by volume ethanol solution, allowed to stand for 6 hours, filtered through a 120-mesh filter to remove the precipitate, and the collected filtrate was concentrated and freeze-dried at -10°C to obtain the kudzu flower alcohol extract.

[0076] Example 10

[0077] An HMGB3 inhibitor is provided, comprising pentagalloylglucose and an alcohol extract of kudzu flower in a mass ratio of 1:2. The HMGB3 inhibitor is obtained by mixing the materials. The alcohol extract of kudzu flower is prepared according to the method of Example 4.

[0078] Example 11

[0079] An HMGB3 inhibitor is provided, comprising a combination of pentagalloylglucose and an alcohol extract of kudzu flower in a mass ratio of 1:3. The HMGB3 inhibitor is obtained by mixing the materials. The alcohol extract of kudzu flower is prepared according to the method of Example 4.

[0080] Example 12

[0081] An HMGB3 inhibitor is provided, comprising pentagalloylglucose and an alcohol extract of kudzu flower in a mass ratio of 1:4. The HMGB3 inhibitor is obtained by mixing the materials. The alcohol extract of kudzu flower is prepared according to the method of Example 4.

[0082] The effects of the present invention will be described below in conjunction with some experiments.

[0083] Study 1: HMGB3 is highly expressed in esophageal squamous cell carcinoma and is associated with poor prognosis

[0084] The research results can be found in Figure 1 and Figure 2 , Figure 1 The HMGB3 immunohistochemical staining IHC score of adjacent tissues and cancerous tissues of esophageal squamous cell carcinoma is Figure 2 Kaplan-Meier survival curves were drawn based on HMGB3 expression levels, and the log-rank test showed P < 0.01. The results showed that HMGB3 was significantly overexpressed in esophageal squamous cell carcinoma, and high expression of HMGB3 was associated with poor prognosis of esophageal squamous cell carcinoma.

[0085] Experiment 2: Expression of HMGB3 in different cells

[0086] Test cells: human esophageal cancer cell lines EC109, EC9706, ECA109, and normal human esophageal cells HET-1A.

[0087] Experimental Methods: Human esophageal cancer cells EC109 were subcutaneously transplanted into immunodeficient mice to establish the esophageal squamous cell carcinoma mouse model EC109-EO. Human esophageal cancer cells EC9706 were subcutaneously transplanted into immunodeficient mice to establish the esophageal squamous cell carcinoma mouse model EC9706-NC. Human esophageal cancer cells ECA109 were subcutaneously transplanted into immunodeficient mice to establish the esophageal cancer mouse model ECA109-EO. Human esophageal normal cells HET-1A were subcutaneously transplanted into immunodeficient mice to serve as a healthy control group.

[0088] Cancer tissue blocks were taken and WB assay was performed using β-actin as an internal reference. Figure 3 , Figure 3 In the data, HET-1A is the result of the healthy control group, human esophageal cancer cell EC9706 is the result of the esophageal squamous cell carcinoma mouse model EC9706-NC, EC109 is the result of the esophageal squamous cell carcinoma mouse model EC109-EO, and human esophageal cancer cell ECA109 is the result of the esophageal cancer mouse model ECA109-EO. Figure 3 It can be seen that the expression level of HMGB3 in the healthy control group is the lowest, while the expression levels in each model group are high, and the band brightness is obvious.

[0089] Experiment 3, Transwell cell migration / invasion assay 1

[0090] The experimental subjects were human esophageal cancer cells EC9706, and the experimental results are shown in Figure 4 , Figure 4 In the study, LV-control was the HMGB3 knockout group, and LV-HMGB3 was the HMGB3 expression group (non-knockout). The results showed that HMGB3 knockout inhibited the invasion and metastasis of EC9706 cells. Conversely, increased HMGB3 expression stimulated the invasion and metastasis of esophageal cancer cells.

[0091] Experiment 4: Inhibition of HMGB3 Expression

[0092] Test cells: human esophageal cancer cell lines EC109, EC9706, ECA109, and normal human esophageal cells HET-1A.

[0093] Experimental Methods: Human esophageal cancer cells EC109 were subcutaneously transplanted into immunodeficient mice to establish the esophageal squamous cell carcinoma mouse model EC109-EO. Human esophageal cancer cells EC9706 were subcutaneously transplanted into immunodeficient mice to establish the esophageal squamous cell carcinoma mouse model EC9706-NC. Human esophageal cancer cells ECA109 were subcutaneously transplanted into immunodeficient mice to establish the esophageal cancer mouse model ECA109-EO. Human esophageal normal cells HET-1A were subcutaneously transplanted into immunodeficient mice to serve as a healthy control group.

[0094] Test treatment and results:

[0095] (1) Esophageal squamous cell carcinoma mouse model EC109-EO, esophageal squamous cell carcinoma mouse model EC109-NC, or esophageal squamous cell carcinoma mouse model ECA109-EO were treated with 10 μg / L PGG at a dose of 100 μL / kg mouse. Healthy mice served as controls. β-ACTIN was used as an internal reference, and WB assays were performed to measure HMGB3 expression. Results are shown in Figure 5 The results showed that PGG treatment could inhibit HMGB3 expression.

[0096] (2) The esophageal squamous cell carcinoma mouse model EC109-EO, esophageal squamous cell carcinoma mouse model EC109-NC, or esophageal squamous cell carcinoma mouse model ECA109-EO were treated with the alcohol extract of Pueraria lobata. The alcohol extract of Pueraria lobata was prepared according to the method of Example 4 and its concentration was 10 μg / L. β-ACTIN was used as an internal reference, and WB assay was performed to test the expression of HMGB3. The results are shown in Table 2. Figure 6 The results showed that the alcohol extract of Pueraria lobata could inhibit the expression of HMGB3.

[0097] In addition, the esophageal squamous cell carcinoma mouse model EC109-EO was treated with the HMGB3 inhibitors of Examples 4 and 10. The dosage of the HMGB3 inhibitor was 100 μL / kg mouse, and the concentration of the HMGB3 inhibitor was 10 μg / L. Using β-ACTIN as an internal reference, a WB assay was performed to test the expression of HMGB3. The results are shown in Figure 7 The results showed that the HMGB3 inhibitors of Examples 4 and 10 could inhibit HMGB3 expression. Example 10 is a combined HMGB3 inhibitor with a more significant inhibitory effect.

[0098] Experiment 5: In vitro inhibition of esophageal squamous cell carcinoma cells

[0099] Test cells: human esophageal cancer cell lines EC109, EC9706, ECA109, and normal human esophageal cells HET-1A.

[0100] When the cells were cultured to 85% confluency, the cells were treated with drugs. After 5 days of incubation, the cell growth inhibition rate was calculated compared to that before drug treatment. The drug addition settings were as follows:

[0101] Blank control: every 10 6 Add 100 μL of 0.9% saline to each EC109 cell.

[0102] Positive control: every 10 6100 μL of 10 μg / L DMSO (dimethyl sulfoxide) was added to each EC109 cell.

[0103] Example 1 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 1 at a concentration of 10 μg / L was added to each EC109 cell.

[0104] Example 2 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 2 at a concentration of 10 μg / L was added to each EC109 cell.

[0105] Example 3 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 3 at a concentration of 10 μg / L was added to each EC109 cell.

[0106] Example 4 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 4 at a concentration of 10 μg / L was added to each EC109 cell.

[0107] Example 5: Every 10 6 100 μL of the HMGB3 inhibitor of Example 5 at a concentration of 10 μg / L was added to each EC109 cell.

[0108] Example 6: Every 10 6 100 μL of the HMGB3 inhibitor of Example 6 at a concentration of 10 μg / L was added to each EC109 cell.

[0109] Example 7: Every 10 6 100 μL of the HMGB3 inhibitor of Example 7 at a concentration of 10 μg / L was added to each EC109 cell.

[0110] Example 8: Every 10 6 100 μL of the HMGB3 inhibitor of Example 8 at a concentration of 10 μg / L was added to each EC109 cell.

[0111] Example 9: Every 10 6 100 μL of the HMGB3 inhibitor of Example 9 at a concentration of 10 μg / L was added to each EC109 cell.

[0112] Example 10 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 10 at a concentration of 10 μg / L was added to each EC109 cell.

[0113] Example 11 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 11 at a concentration of 10 μg / L was added to each EC109 cell.

[0114] Example 12: Every 10 6 100 μL of the HMGB3 inhibitor of Example 12 at a concentration of 10 μg / L was added to each EC109 cell.

[0115] Experimental method: The cell growth inhibition rate was determined by MTT assay.

[0116] Result processing: Each experiment was set up with 3 parallel treatments, and the growth inhibition rate results were averaged.

[0117] The results in Table 1 show that Examples 1 through 12 demonstrated excellent inhibition rates against various cancer cells, with Examples 10 through 12 showing particularly significant effects. Damage to normal human esophageal HET-1A cells was minimal. Although DMSO was used as a positive control, the inhibitory effect was poor due to the low concentration.

[0118] Table 1 Cell growth inhibition rate results

[0119]

[0120]

[0121] Experiment 6: Transwell cell migration / invasion assay 2

[0122] The experimental subject is human esophageal cancer cell EC9706.

[0123] Blank control: every 10 6 Add 100 μL of 0.9% saline to each EC109 cell.

[0124] Positive control: every 10 6 Add 100 μL of 10 μg / L DMSO to each EC109 cell.

[0125] Example 1 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 1 at a concentration of 10 μg / L was added to each EC109 cell.

[0126] Example 2 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 2 at a concentration of 10 μg / L was added to each EC109 cell.

[0127] Example 3 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 3 at a concentration of 10 μg / L was added to each EC109 cell.

[0128] Example 4 Group: Every 106 100 μL of the HMGB3 inhibitor of Example 4 at a concentration of 10 μg / L was added to each EC109 cell.

[0129] Example 5: Every 10 6 100 μL of the HMGB3 inhibitor of Example 5 at a concentration of 10 μg / L was added to each EC109 cell.

[0130] Example 6: Every 10 6 100 μL of the HMGB3 inhibitor of Example 6 at a concentration of 10 μg / L was added to each EC109 cell.

[0131] Example 7: Every 10 6 100 μL of the HMGB3 inhibitor of Example 7 at a concentration of 10 μg / L was added to each EC109 cell.

[0132] Example 8: Every 10 6 100 μL of the HMGB3 inhibitor of Example 8 at a concentration of 10 μg / L was added to each EC109 cell.

[0133] Example 9: Every 10 6 100 μL of the HMGB3 inhibitor of Example 9 at a concentration of 10 μg / L was added to each EC109 cell.

[0134] Example 10 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 10 at a concentration of 10 μg / L was added to each EC109 cell.

[0135] Example 11 Group: Every 10 6 100 μL of the HMGB3 inhibitor of Example 11 at a concentration of 10 μg / L was added to each EC109 cell.

[0136] Example 12: Every 10 6 100 μL of the HMGB3 inhibitor of Example 12 at a concentration of 10 μg / L was added to each EC109 cell.

[0137] Results processing: Each experiment was set up for 3 parallel treatments, and the cell migration and invasion results were averaged. The results in Table 2 show that the inhibitors of Example 1 to Example 12 can effectively inhibit the migration and invasion of esophageal cancer cells. Although DMSO was used as the positive control, the inhibitory effect was poor due to the low concentration.

[0138] Table 2 Cell migration and invasion results

[0139] drug Migration Results Invasion results Blank control 100±7 142±6 Positive control 79±5 116±6 Example 1 group 50±3 71±4 Example 2 group 71±4 85±3 Example 3 group 65±5 71±6 Example 4 Group 51±6 60±3 Example 5 group 50±3 60±4 Example 6 50±2 58±3 Example 7 Group 51±2 59±2 Example 8 Group 52±2 60±3 Example 9 Group 62±3 70±4 Example 10 Group 42±2 52±3 Example 11 Group 40±3 50±2 Example 12 Group 41±4 50±1

[0140] It should be noted that when numerical ranges are involved in the present invention, it should be understood that both 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 preferred embodiments. Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they understand the basic inventive concept, and such changes and modifications fall within the scope of the present invention.

[0141] Obviously, those skilled in the art may 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 intended to include these modifications and variations.

Claims

1. Use of an HMGB3 inhibitor in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The HMGB3 inhibitor is an alcohol extract of Pueraria lobata flower, or a combination of pentagalloylglucose and an alcohol extract of Pueraria lobata flower; When the esophageal cancer cell proliferation and migration drug is composed of pentagalloylglucose and the alcohol extract of Pueraria lobata flower, the mass ratio of the two is 1:2~4; The alcohol extract of Pueraria lobata flower is prepared according to the following method: Fresh kudzu flower was taken, washed, dried, and crushed into 60-100 mesh powder, then mixed with 60% ethanol solution by volume, allowed to stand for 6-12 hours, filtered to remove the precipitate, and the collected filtrate was concentrated and freeze-dried to obtain the kudzu flower alcohol extract.

2. Use of the HMGB3 inhibitor according to claim 1 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The drying is carried out at 30°C to 40°C.

3. Use of the HMGB3 inhibitor according to claim 1 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The material-liquid ratio of powder to 60% volume fraction ethanol solution is 1kg:8L.

4. Use of the HMGB3 inhibitor according to claim 1 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The freeze-drying condition is to freeze-dry at -20℃.

5. Use of the HMGB3 inhibitor according to claim 1 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The kudzu flowers are the unopened flower buds of the kudzu vine.

6. Use of the HMGB3 inhibitor according to claim 1 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The esophageal cancer cells are one or more of human esophageal cancer cells EC109, human esophageal cancer cells EC9706, and human esophageal cancer cells ECA109.

7. Use of the HMGB3 inhibitor according to claim 1 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: It is also compounded with pharmaceutically acceptable excipients.

8. Use of the HMGB3 inhibitor according to claim 7 in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer cells, characterized in that: The esophageal cancer cell proliferation and migration drug has an HMGB3 inhibitor as the only active ingredient.

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