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Application of niclosamide in treatment or prevention of melanoma

A technology for niclosamide and melanoma, applied in the application field of niclosamide in the treatment or prevention of melanoma, can solve the problems of high risk, slow progress of drug resistance mechanism research, unknown biological safety, etc. The effect of biosecurity

Inactive Publication Date: 2018-11-16
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Even so, vemurafenib will produce severe drug resistance after a period of treatment, which limits the wide application of vemurafenib in the field of melanoma treatment
[0005] Therefore, the following challenges still exist in the chemotherapy treatment of melanoma: first, more effective chemotherapy drugs need to be developed urgently; second, new drug research cycle is long, risky, unknown biological safety and other issues limit its development, and the research on drug resistance mechanism is slow

Method used

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  • Application of niclosamide in treatment or prevention of melanoma
  • Application of niclosamide in treatment or prevention of melanoma
  • Application of niclosamide in treatment or prevention of melanoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Niclosamide is cytotoxic to melanoma cell A375 and can inhibit its proliferation

[0059] 1.1 Trypan blue staining live cell counting method

[0060] The A375 cell lines treated with different concentrations of niclosamide (0, 1, 2, 4, 8, 16 μmol / L) were observed under an optical microscope at three time points of 24 / 48 / 72 hours, and it was found that with the concentration increased, the number of cells at each time point was significantly reduced, and in the 16μM concentration group at 72h, a large number of cells died (see figure 1 ). Detected by trypan blue staining and viable cell counting method, consistent with the results observed under the microscope, it was found that the A375 cells after drug treatment were significantly reduced compared with the control group, and the higher the drug concentration, the less the number of remaining viable cells (P figure 2 ).

[0061] 1.2CCK-8 method

[0062] Using the CCK-8 method, the A375 cell lines treated with differe...

Embodiment 2

[0066] Niclosamide can inhibit the migration ability of melanoma cell A375

[0067] 2.1 Cell scratch experiment

[0068] The migration ability of A375 cell line was tested by cell scratch test, and ImageJ 1.48v was used for image analysis. The results showed that the scratch closure speed of the niclosamide treatment group was significantly slower than that of the control group (P Figure 5 ).

[0069] 2.2 Transwell method

[0070] Transwell method was used to detect the invasion ability of A375 cell line, and after crystal violet staining, ImageJ 1.48v was used for image analysis. The results showed that after 4 / 8 / 12h of A375 cells treated with different concentrations of niclosamide (0, 2, 4, 8 μmol / L), the cell membrane penetration ability of the drug treatment group was significantly lower than that of the control group (P Image 6 ).

[0071] The cells stained with crystal violet were eluted with 10% acetic acid and then read with a spectrophotometer. The results showed...

Embodiment 3

[0073] 3.1 Hoechst staining

[0074] Hoechst staining was used to detect the effect of niclosamide on the apoptosis of A375 cell line. A375 cells were treated with different concentrations of niclosamide (0, 1, 2, 4, 8, 16 μmol / L) for 4 / 8 / 12 hours, and the cells were collected for Hoechst staining, photographed and analyzed.

[0075] The results showed that, compared with the control group, the number of apoptotic cells observed under the high-power lens field after niclosamide treated cells for 8 hours was significantly increased, which was statistically significant (P Figure 8 ), and the higher the drug concentration, the more apoptotic cells were observed.

[0076] 3.2 Analysis of cell apoptosis by flow cytometry

[0077] Cell apoptosis analysis was carried out by flow cytometry, and the ability of niclosamide to promote apoptosis of A375 cell line was verified again. Treat A375 cells with different concentrations of niclosamide (0, 2, 4, 8 μmol / L) for 4 / 8 / 12 hours, colle...

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Abstract

The invention discloses application of niclosamide in treatment or prevention of melanoma, and relates to the field of melanoma treatment or prevention drugs. A research of the invention finds that the niclosamide has an obvious anti-tumor effect; the niclosamide can effectively inhibit multiplication of the melanoma; the niclosamide can inhibit migration and invasion of the melanoma; the niclosamide can promote apoptosis of the melanoma; and the research further finds that the niclosamide agent can improve the activity of tyrosinase in melanoma cells and promote secretion of melanin, and indicates that the niclosamide agent can promote terminal differentiation of the melanoma cells. The research of the invention indicates that the niclosamide has a wide application value in the field of treatment or prevention of the melanoma and the like and has a prospect of serving as a melanoma resisting drug.

Description

technical field [0001] The present invention relates to the field of medicine for treating or preventing melanoma, in particular, relates to the application of niclosamide in treating or preventing melanoma. Background technique [0002] Melanoma has a high degree of malignancy, early metastasis, and a high mortality rate, accounting for 80% of skin cancers. Very early-stage melanoma can be cured with surgery. Since the early symptoms of melanoma are not obvious, most patients are already in the middle and late stage when they are discovered. They are not sensitive to radiotherapy and mainly rely on chemotherapy, but have poor sensitivity to chemotherapy drugs. Drug resistance is the bottleneck of current melanoma treatment. Therefore, the treatment of melanoma needs to be carried out by developing new drugs to suppress tumors or researching the mechanism of drug resistance. However, the research cycle of new drugs is long, the risk is high, and the biological safety is un...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/167A61P35/00
CPCA61K31/167A61P35/00
Inventor 邓芳张志辉张浩辛浩然
Owner ARMY MEDICAL UNIV
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