HMGB3 inhibitor and application of HMGB3 inhibitor in preparation of drugs for inhibiting proliferation and migration of esophageal cancer cells

Abstract

The invention belongs to the technical field of esophageal cancer inhibitors, and particularly relates to an HMGB3 inhibitor and application of the HMGB3 inhibitor in preparation of drugs for inhibiting esophageal cancer cell proliferation and migration. It is found that compared with para-carcinoma tissue, the expression of HMGB3 in esophageal squamous carcinoma tissue is remarkably increased, so that proliferation and migration of esophageal cancer cells are inhibited by inhibiting the expression of HMGB3. The HMGB3 inhibitor provided by the invention can inhibit expression of HMGB3 in a targeted manner and inhibit proliferation and migration of esophageal squamous carcinoma cells.

Description

Technical Field

[0001] The present invention belongs to the technical field of esophageal cancer inhibitors, and in particular relates to an HMGB3 inhibitor and an application thereof 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, and esophageal squamous cell carcinoma is a specific histopathological classification. Clinically, most esophageal cancers are squamous cell carcinomas, and a few are adenocarcinomas. The full name of esophageal squamous cell carcinoma is esophageal squamous cell carcinoma, abbreviated as ESCC, which refers to esophageal malignant tumors caused by dysplasia of esophageal squamous epithelium. Early clinical symptoms include swallowing discomfort, foreign body sensation, pain behind the sternum, etc., and late 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 the structure of DNA 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 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 preparations that act on the therapeutic targets of esophageal cancer is of great significance for conquering esophageal cancer. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides an HMGB3 inhibitor and its use in preparing a drug for inhibiting the proliferation and migration of esophageal cancer cells. The HMGB3 inhibitor acts on the therapeutic target HMGB3 of 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 a HMGB3 inhibitor, wherein the HMGB3 inhibitor is pentagalloyl glucose, an alcohol extract of kudzu 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] Take fresh kudzu flower, wash, dry, crush into 60-100 mesh powder, then mix with 60%-70% ethanol solution, let stand for 6h-12h, filter to remove precipitation, concentrate the collected filtrate, freeze-dry to obtain kudzu flower alcohol extract.

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

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

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

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

[0014] The second object of the present invention is to provide an application of an HMGB3 inhibitor in the preparation of a drug for inhibiting the proliferation and migration of esophageal cancer. The present invention clearly found that the expression of HMGB3 in esophageal squamous cell carcinoma tissues was significantly increased compared with adjacent adjacent tissues, that is, the expression level of HMGB3 was highly correlated with esophageal cancer, and HMGB3 could be used as a therapeutic target for esophageal cancer. By inhibiting the expression of HMGB3, the proliferation and migration of esophageal cancer cells could be inhibited.

[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, talcum powder, and micro-powder 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 is deemed to be 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, which is pentagalloylglucose, an alcohol extract of kudzu flower, or a combination of the two. The HMGB3 inhibitor can target and inhibit the expression of HMGB3, and inhibit the proliferation and migration of esophageal cancer cells. It is a new preparation that acts on the therapeutic target of esophageal cancer, and is of great significance for conquering esophageal cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the IHC score of HMGB3 immunohistochemical staining of adjacent tissues and cancer tissues of esophageal squamous cell carcinoma.

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

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

[0031] Figure 4The results of Transwell cell migration / invasion assay 1.

[0032] Figure 5 The results of WB test showing that PGG inhibited the expression of HMGB3.

[0033] Figure 6 The results of a WB experiment showing that the alcohol extract of Pueraria lobata flower inhibited 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 in conjunction with 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, inhibiting the proliferation and migration of esophageal cancer cells. After combining multiple substances, the effect of inhibiting the expression of HMGB3 and stimulating the expression of HMGB3 is more obvious.

[0040] Based on the same inventive concept, the present invention provides an application 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 in the digestive tract tumors 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] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0047] Take fresh kudzu flower, i.e. the open flower of sweet kudzu vine, wash to remove dust and other impurities, keep the pedicel and petals, dry at 80°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, freeze-dry at -20°C to obtain the kudzu flower alcohol extract, and store at -20°C for future use.

[0048] Example 3

[0049] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0051] Take fresh kudzu flowers, i.e. unopened buds of sweet kudzu vine, wash to remove dust and other impurities, retain the pedicels and petals, dry at 80°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, freeze-dry at -20°C to obtain the kudzu flower alcohol extract, and store at -20°C for later use.

[0052] Example 4

[0053] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0055] Take fresh kudzu flowers, i.e. unopened 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, freeze-dry at -20°C to obtain the kudzu flower alcohol extract, and store at -20°C for later use.

[0056] Example 5

[0057] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

[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 sweet kudzu vine, wash to remove dust and other impurities, retain the pedicels and petals, dry at 40°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, freeze-dry at -20°C to obtain the kudzu flower alcohol extract, and store at -20°C for later use.

[0060] Example 6

[0061] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0063] 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 100-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, freeze-dry at -20°C to obtain the kudzu flower alcohol extract, and store at -20°C for later use.

[0064] Example 7

[0065] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0067] Take fresh kudzu flowers, i.e. unopened 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 70% 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.

[0068] Example 8

[0069] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0071] Take fresh kudzu flowers, i.e. unopened 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 12 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.

[0072] Example 9

[0073] A HMGB3 inhibitor, which is an alcohol extract of Pueraria lobata flower.

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

[0075] Take fresh kudzu flowers, i.e. unopened 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 -10°C to obtain the kudzu flower alcohol extract.

[0076] Example 10

[0077] A HMGB3 inhibitor is provided, wherein the HMGB3 inhibitor is composed of pentagalloylglucose and an alcohol extract of kudzu flower in a mass ratio of 1:2. The materials are mixed to obtain the HMGB3 inhibitor. The alcohol extract of kudzu flower is prepared according to the method of Example 4.

[0078] Embodiment 11

[0079] A HMGB3 inhibitor is provided, wherein the HMGB3 inhibitor is composed of pentagalloylglucose and an alcohol extract of kudzu flower in a mass ratio of 1:3. The materials are mixed to obtain the HMGB3 inhibitor. The alcohol extract of kudzu flower is prepared according to the method of Example 4.

[0080] Example 12

[0081] A HMGB3 inhibitor is provided, wherein the HMGB3 inhibitor is composed of pentagalloylglucose and an alcohol extract of kudzu flower in a mass ratio of 1:4. The materials are mixed to obtain the HMGB3 inhibitor. The alcohol extract of kudzu flower is prepared according to the method of Example 4.

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

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

[0084] The results of the study can be found in Figure 1 and Figure 2 , Figure 1 The IHC score of HMGB3 immunohistochemical staining of adjacent tissues and cancer tissues of esophageal squamous cell carcinoma. Figure 2 Kaplan-Meier survival curve was drawn according to the expression level of HMGB3, and the log-rank test 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 line EC109, human esophageal cancer cell line EC9706, human esophageal cancer cell line ECA109, and human esophageal normal cell line HET-1A.

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

[0088] Cancer tissue blocks were taken and WB test was performed with β-actin as 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. 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 test subjects were human esophageal cancer cells EC9706, and the test results are shown in Figure 4 , Figure 4 In the experiment, LV-control was the HMGB3 knockout group, and LV-HMGB3 was the HMGB3 expression group (not knocked out). The results showed that HMGB3 knockout inhibited the invasion and metastasis of EC9706 cells. On the contrary, the increase of HMGB3 expression stimulated the invasion and metastasis of esophageal cancer cells.

[0091] Experiment 4: Experiment on inhibiting HMGB3 expression

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

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

[0094] Test treatment and results:

[0095] (1) PGG was used to treat 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 at a concentration of 10 μg / L. The PGG dosage was 100 μL / kg mouse. Healthy mice were used as controls. β-ACTIN was used as an internal reference, and WB experiments were performed to test the expression of HMGB3. The results are shown in Figure 5 The results showed that PGG treatment could inhibit HMGB3 expression.

[0096] (2) The alcohol extract of Pueraria lobata was used to treat the esophageal squamous cell carcinoma mouse model EC109-EO, the esophageal squamous cell carcinoma mouse model EC109-NC or the esophageal carcinoma mouse model ECA109-EO, respectively. The alcohol extract of Pueraria lobata was used at a dosage of 100 μL / kg mouse. 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 a WB test was performed to test the expression of HMGB3. The results are shown in Figure 6 The results showed that the alcohol extract of Pueraria lobata flower 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 Example 4 and Example 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 test was performed to test the expression of HMGB3. The results are shown in Figure 7 The results show that the HMGB3 inhibitors of Example 4 and Example 10 can 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 line EC109, human esophageal cancer cell line EC9706, human esophageal cancer cell line ECA109, and human esophageal normal cell line HET-1A.

[0100] 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:

[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 Group: 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 Group: 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 Group: 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 Group: 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 Group: 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 Group: 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 test was set up with 3 parallel treatments, and the growth inhibition rate results were averaged.

[0117] The results in Table 1 show that Example 1 to Example 12 have a good inhibition rate on various cancer cells, among which Example 10 to Example 12 have a particularly obvious effect. However, the damage to normal human esophageal normal cells HET-1A is very small. Although DMSO was used as the positive control, the inhibition effect was poor due to the low concentration.

[0118] Table 1 Cell growth inhibition rate results

[0119]

[0120]

[0121] Experiment VI: Transwell cell migration / invasion assay II

[0122] The experimental subject was 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 Group: 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 Group: 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 Group: 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 Group: 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 Group: 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 Group: 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 test 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 Group 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 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.

[0141] 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. A HMGB3 inhibitor, characterized in that The HMGB3 inhibitor is pentagalloyl glucose, alcohol extract of Pueraria lobata flower, or a combination of the two; 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.

2. The HMGB3 inhibitor according to claim 1, characterized in that The alcohol extract of Pueraria lobata flower was prepared according to the following method: Take fresh kudzu flower, wash, dry, crush into 60-100 mesh powder, then mix with 60%-70% ethanol solution, let stand for 6h-12h, filter to remove precipitation, concentrate the collected filtrate, freeze-dry to obtain kudzu flower alcohol extract.

3. The HMGB3 inhibitor according to claim 2, characterized in that The drying is carried out at 30°C to 40°C.

4. The HMGB3 inhibitor according to claim 2, characterized in that The solid-liquid ratio of the powder to the ethanol solution with a volume fraction of 60% to 70% is 1kg:8L.

5. The HMGB3 inhibitor according to claim 2, characterized in that The freeze-drying condition is to freeze-dry at -20°C.

6. The HMGB3 inhibitor according to claim 2, characterized in that The kudzu flowers are unopened flower buds of kudzu or unopened flower buds of sweet kudzu.

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.

8. The use according to claim 7, 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.

9. The use according to claim 7, characterized in that: The drug is obtained by compounding the HMGB3 inhibitor with pharmaceutically acceptable excipients.

10. The use according to claim 9, characterized in that: The drug contains an HMGB3 inhibitor as the only active ingredient.

Images