Application of mebendazole in preparation of medicine with osteosarcoma prevention and / or treatment effect

Abstract

The invention relates to the technical field of medicinal chemistry, in particular to application of mebendazole in preparation of a medicine with an osteosarcoma prevention and / or treatment effect. The invention provides a brand new application of mebendazole. Researches show that mebendazole can significantly inhibit proliferation, invasion and migration capabilities of osteosarcoma cells in vitro, can inhibit in-situ growth of osteosarcoma cells in vivo, and significantly reduces the number and area of pulmonary metastatic nodules; therefore, mebendazole serving as an active ingredient is used for preparing the medicine with the effect of preventing and / or treating osteosarcoma, especially the medicine with the effect of preventing and / or treating metastasis, and has a wide application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical chemistry, and in particular to the application of mebendazole in preparing a medicine capable of preventing and / or treating osteosarcoma. Background Art

[0002] Osteosarcoma (OS) is a primary malignant bone tumor that originates from primitive mesenchymal tissue and is common in children and adolescents. Currently, the treatment of osteosarcoma mainly uses surgery and chemotherapy based on cisplatin, cyclophosphamide, doxorubicin and methotrexate. Chemotherapy can increase the survival rate of osteosarcoma patients to 60-70% within 5 years, but it cannot prevent its metastasis. At present, there is a lack of preventive or therapeutic drugs for metastatic osteosarcoma in clinical practice. Therefore, the development of drugs to prevent or treat metastatic osteosarcoma has important clinical application value.

[0003] Mebendazole is a broad-spectrum antiparasitic drug. Studies have shown that mebendazole inhibits the function of microtubules in worms and animal cells by inhibiting β-tubulin polymerization, resulting in reduced secretory vesicle transport, reduced glucose uptake and increased glycogenolysis. Preclinical studies have found that mebendazole can inhibit the progression of melanoma, adrenocortical carcinoma, medulloblastoma, colorectal cancer, head and neck squamous cell carcinoma, breast cancer and acute lymphoblastic leukemia by inhibiting HIF1 / 2, USP5 and VEGFR2, degrading integrin 4, MYB and c-Maf.

[0004] However, the application and research of mebendazole in osteosarcoma, especially metastatic osteosarcoma, have not been reported. In addition, melanoma, adrenocortical carcinoma, medulloblastoma, colorectal cancer, head and neck squamous cell carcinoma, breast cancer and acute lymphoblastic leukemia are different cancers from osteosarcoma; most of the drugs that can treat and prevent cancers such as melanoma, adrenocortical carcinoma, medulloblastoma, colorectal cancer, head and neck squamous cell carcinoma, breast cancer and acute lymphoblastic leukemia cannot be used to treat and prevent osteosarcoma. Therefore, those skilled in the art cannot predict whether mebendazole has the effect of inhibiting osteosarcoma cells when mebendazole can inhibit the above-mentioned cancer cells; and it is even more impossible to predict whether mebendazole can inhibit the metastasis of osteosarcoma cells. Summary of the invention

[0005] In order to overcome at least one of the technical problems existing in the prior art, the present invention provides the use of mebendazole in preparing a medicament having the function of preventing and / or treating osteosarcoma.

[0006] The present invention firstly provides the use of mebendazole in preparing a medicine capable of preventing and / or treating osteosarcoma.

[0007] Preferably, the osteosarcoma is metastatic osteosarcoma.

[0008] Preferably, mebendazole prevents and / or treats metastatic osteosarcoma by inhibiting the migration of osteosarcoma cells.

[0009] Preferably, mebendazole prevents and / or treats metastatic osteosarcoma by inhibiting the invasive ability of osteosarcoma cells.

[0010] Preferably, mebendazole achieves the effect of preventing and / or treating metastatic osteosarcoma by inhibiting the metastasis of osteosarcoma cells.

[0011] Preferably, the metastatic osteosarcoma is pulmonary metastatic osteosarcoma.

[0012] Preferably, mebendazole achieves the effect of preventing and / or treating pulmonary metastatic osteosarcoma by inhibiting the pulmonary metastasis of osteosarcoma cells.

[0013] The present invention also provides the use of mebendazole in preparing a medicine having the function of inhibiting the proliferation of osteosarcoma cells.

[0014] The present invention also provides the use of mebendazole in preparing a medicine having the function of inhibiting the migration of osteosarcoma cells.

[0015] The present invention also provides the use of mebendazole in preparing a medicine capable of inhibiting the invasion ability of osteosarcoma cells.

[0016] The present invention also provides the use of mebendazole in preparing a medicine having an anti-osteosarcoma metastasis effect.

[0017] Preferably, the osteosarcoma metastasis specifically refers to osteosarcoma lung metastasis.

[0018] Preferably, the drug contains mebendazole or a physiologically acceptable salt of mebendazole as an active ingredient, and also contains a pharmaceutically acceptable administration carrier and / or excipient.

[0019] Preferably, the physiologically acceptable salts include various acidic salts formed with organic acids and inorganic acids and various basic salts formed with organic bases and inorganic bases.

[0020] Beneficial effects: The present invention provides a new application of mebendazole; studies have shown that mebendazole can significantly inhibit the proliferation, invasion and migration of osteosarcoma cells in vitro, inhibit the in situ growth of osteosarcoma cells in vivo, and significantly reduce the number and area of ​​pulmonary metastatic nodules; therefore, mebendazole is used as an active ingredient in the preparation of drugs with the effect of preventing and / or treating osteosarcoma, especially in the preparation of drugs with the effect of preventing and / or treating metastatic osteosarcoma, especially in the preparation of drugs with the effect of resisting osteosarcoma pulmonary metastasis, which has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The inhibitory effect of mebendazole on the migration ability of 143B and SJSA-1 (*P<0.05vs DMSO control group).

[0022] Figure 2 The inhibitory effect of mebendazole on the invasion ability of 143B and SJSA-1 cells (*P<0.05, **P<0.001 and ***P<0.001 vs DMSO control group). DETAILED DESCRIPTION

[0023] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.

[0024] The chemical structure of mebendazole involved in the embodiments of the present invention is as follows:

[0025]

[0026] Example 1 Inhibitory effect of mebendazole on the proliferation of human osteosarcoma cells

[0027] Experimental method: HOS, MNNG / HOS, 143B, SJSA-1 and NOB (normal human osteoblasts) cells in the logarithmic growth phase were digested and seeded into 96-well plates at a seeding density of 3×10 3 cells / well. After overnight culture, the culture medium was removed, and culture medium containing different drug concentrations was added to each well, and an equal volume of DMSO was used as a negative control. After 48 hours of cell culture, the culture medium was removed, and culture medium containing CCK8 (1:100 dilution) was added to each well. The cells were incubated in a cell culture incubator for 2 hours, and the absorbance at 450nm was measured using an ELISA reader. Cell survival rate % = (average OD value of the drug treatment group - average OD value of the blank well) / (average OD value of the control group - average OD value of the blank well) × 100%.

[0028] Experimental results: Low concentrations of mebendazole can inhibit the proliferation of osteosarcoma cells, but have little effect on the proliferation of normal human osteoblasts (see Table 1).

[0029] Table 1. Inhibitory effect of mebendazole on the proliferation of osteosarcoma cells

[0030]

[0031]

[0032] Example 2 Inhibitory effect of mebendazole on the migration ability of 143B and SJSA-1 cells

[0033] Experimental method: Digest and centrifuge the cells in the logarithmic growth phase, resuspend the cells, and adjust the cell suspension concentration to 4×10 5 / ml. Use tweezers to place the sterile Ibidi plug in a 12-well plate, add 70 μL of cell suspension on both sides of the plug, and incubate overnight. Remove the plug, wash with PBS, add culture medium containing 1% FBS and different concentrations of mebendazole, and take pictures at 0h and 24h.

[0034] Experimental results: Compared with the DMSO group, mebendazole can significantly inhibit the migration ability of 143B and SJSA-1 cells at a non-toxic dose (see Figure 1 ).

[0035] Example 3 Inhibitory effect of mebendazole on the invasion ability of 143B and SJSA-1 cells

[0036] Experimental method: The cells in the logarithmic growth phase were digested, centrifuged, resuspended in culture medium, and the culture medium was adjusted to 1×10 4 Cells were inoculated into the upper chamber of a Transwell nest with a pore size of 8.0 μm and 20 μL of 2.5 mg / mL Matrigel matrix gel (Lambolide) at a volume of 100 μL of cell suspension, and 500 μL of complete culture medium containing different concentrations of tobendazole was added to the lower chamber. After 48 hours of incubation in the incubator, the culture medium in the upper and lower chambers was removed, the cells were fixed with 4% paraformaldehyde for 30 minutes, rinsed with PBS, and 500 μL of 0.1% crystal violet solution was added to the lower chamber for staining. The chamber was rinsed with PBS, and the cells in the upper chamber of the nest were wiped off with a cotton swab and dried. Five fields of view were randomly selected from each chamber for photography, and the number of migrated cells was counted using ImageJ.

[0037] Experimental results: Compared with the DMSO group, non-toxic doses of mebendazole could significantly inhibit the invasion ability of 143B and SJSA-1 cells (see Figure 2 ).

[0038] Example 4 Inhibitory effect of mebendazole on the growth and lung metastasis of 143B and SJSA-1 cells orthotopically transplanted into nude mice

[0039] Experimental method: 143B and SJSA-1 cells in the logarithmic growth phase were collected, rinsed 3 times with PBS, centrifuged, resuspended with PBS, and the cell density was adjusted to 4×10 7 Use an islet syringe to draw up 50 μL of cell suspension and inject it into the proximal tibia of 4-week-old male BALB / C nude mice. When the tumor volume grows to ~80mm 3About 10 minutes later, the tumor-bearing mice were randomly divided into a solvent control group and a mebendazole administration group (medium and high dose groups), with 8 mice in each group. Mebendazole was dissolved in sesame oil to make a suspension, which was administered by gavage. The dosages were 50 mg / kg and 100 mg / kg, once a day. The body weight and tumor diameter of the nude mice were recorded. The tumor diameter calculation formula for osteosarcoma is: V = 4 / 3π[1 / 4(a+b)]2, where a and b are the long and short diameters of the tumor perpendicular to the tibia, respectively. After 4 weeks of drug treatment, the mice were killed, and the tumors and lung tissues were dissected. The tumor tissues were weighed and photographed. The tumor and lung tissues were fixed and embedded with 4% paraformaldehyde, sectioned, stained with H&E, and analyzed.

[0040] Experimental results: Mebendazole can inhibit the growth of 143B and SJSA-1 transplanted tumors, with tumor inhibition rates of 34.04%, 55.24% and 37.65%, 49.89%, respectively, and has no significant effect on the body weight of nude mice (Table 2). Mebendazole can significantly reduce the number and area of ​​143B and SJSA-1 cell lung metastases (Table 3).

[0041] Table 2. Inhibitory effect of mebendazole on the growth of 143B and SJSA-1 transplanted tumors

[0042]

[0043] Table 3. Inhibitory effect of mebendazole on lung metastasis of 143B and SJSA-1 transplanted tumors

[0044]

Claims

1. The use of mebendazole in the preparation of a drug for preventing and / or treating osteosarcoma.

2. The use according to claim 1, characterized in that: The osteosarcoma is metastatic osteosarcoma.

3. The use of mebendazole in the preparation of drugs having the effect of inhibiting the proliferation of osteosarcoma cells.

4. The use of mebendazole in the preparation of drugs that inhibit the migration of osteosarcoma cells.

5. The use of mebendazole in the preparation of drugs capable of inhibiting the invasive ability of osteosarcoma cells.

6. The use of mebendazole in the preparation of drugs with anti-osteosarcoma metastasis effect.

7. The use according to any one of claims 1 to 6, characterized in that: The medicine uses mebendazole or a physiologically acceptable salt of mebendazole as an active ingredient, and also contains a pharmaceutically acceptable administration carrier and / or excipient.

8. The use according to claim 7, characterized in that: The physiologically acceptable salts include various acidic salts formed with organic acids and inorganic acids and various basic salts formed with organic bases and inorganic bases.

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