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Application of combined stephanine and autophagy inhibitor for preparing drugs for treating liver cancer

A technology of autophagy inhibitor and fenugreek, applied in the field of medicine, can solve the problems of reducing the effect of anti-tumor drugs, different conclusions, and not too many reports, and achieves simple and economical acquisition methods, wide sources, enhanced The effect of anticancer action

Active Publication Date: 2019-11-26
GUANGZHOU JINAN BIOMEDICINE RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And more and more studies have shown that stephine can reverse the drug resistance of tumors and treat tumors as an adjuvant drug, but because autophagy can promote tumor survival and reduce the effect of anti-tumor drugs, stephine alone inhibits tumors. There are not too many reports on the role of autophagy. There are only a few reports on stephanine and autophagy so far, but the conclusions are not the same.
We discovered the protective mechanism of autophagy on tumor cells, and then combined stephine and autophagy inhibitors to achieve anti-tumor effects, but the combination of stephine and autophagy inhibitors has not been reported yet, and the dose of their combination There is no relevant report

Method used

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  • Application of combined stephanine and autophagy inhibitor for preparing drugs for treating liver cancer
  • Application of combined stephanine and autophagy inhibitor for preparing drugs for treating liver cancer
  • Application of combined stephanine and autophagy inhibitor for preparing drugs for treating liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Weigh stepherine hydrochloride powder with an electronic balance, add it into DMSO to dissolve, make a mother solution with a concentration of 100mM, and then dilute it with DMSO to different concentrations. The MTT method was used to detect the toxicity of stephine hydrochloride to normal liver cells HL-7702 and liver cancer cells SMCC-7721, Huh7, HepG2 cells for 24h and 48h, and the IC of stephine hydrochloride to different liver cancer cells was calculated 50 value.

[0038] Specific method: Normal liver cells HL-7702 were digested with trypsin and inoculated into 96-well plates, and 0.4×10 cells were inoculated in each well. 4 After culturing overnight to adhere to the wall, add 0-100uM stephine hydrochloride to the culture dish, add 10uL MTT (5mg / mL) when culturing for 24h and 48h respectively, incubate at 37°C for 4h and suck off the culture medium and MTT , add DMSO, react on a shaker at room temperature for 30 minutes, measure the absorbance at 570 nm, and then...

Embodiment 2

[0041] On the premise of Example 1, the present invention further verified that stephine hydrochloride does have an effect on the growth of liver cancer cells, and then further carried out scratch, clone formation, TransWell, and flow sorting experiments to detect apoptotic cells.

[0042] The specific method of the scratch test: first draw a black horizontal line on the bottom of the outer surface of the 12-well plate, then digest the Huh7 cells with trypsin and inoculate 20×10 4 Cells were seeded into 12-well plates and cultured overnight. After the cells adhere to the wall, wash it once with PBS, and then use a 200uL pipette tip to draw a horizontal line on the bottom of the inner surface of the 12-well plate, which is perpendicular to the horizontal line on the outer surface, causing cell scratches. After the process is completed, wash with PBS and divide into 3 groups (control group (indicated by control), cisplatin group (indicated by CDDP), stephine hydrochloride group ...

Embodiment 3

[0048] On the basis of Examples 1 and 2, stephine hydrochloride and autophagy inhibitors were used in combination to inhibit liver cancer cells. The liver cancer cell Huh7 was digested with trypsin, and the cell density was adjusted to 20×10 4 cells / mL, add 100uL / well of cell culture medium into a 96-well plate, remove the culture medium after culturing for 12 hours, and use different concentrations of stephine hydrochloride (m=1-1000) and different concentrations of autophagy inhibitor chloroquine (n=1-1000) mixed according to the ratio of 1:7.5, added to the 96-well plate, after culturing for 48 hours, the survival rate of the cells was measured by MTT, and the survival rate of liver cancer cells when combined with stephine hydrochloride and chloroquine alone , CompuSyn software was used to calculate and analyze the combination index of the two drugs. The combined index analysis chart of stephine hydrochloride and autophagy inhibitor combined drug Figure 6 As shown, diffe...

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Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of combined stephanine and an autophagy inhibitor for preparing drugs for treating a liver cancer. Itis found that drug combination of the cepharanthine hydrochloride and the autophagy inhibitor has a good cancer inhibition effect in vitro and vivo, more treatment solid tumor schemes are provided, and two combined drugs are wide in source and are economic and effective. According to the application of the combined stephanine and the autophagy inhibitor for preparing the drugs for treating the liver cancer, it is firstly found that by using the autophagy inhibitor to reinforce the antitumor effect of the cepharanthine hydrochloride in vitro and vivo, the antitumor effect of the cepharanthinehydrochloride in vitro and vivo is verified on a vivo mouse model, and a novel scheme for effectively and feasibly treating a solid tumor is provided.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to the application of Stephanine in combination with an autophagy inhibitor in the preparation of a drug for treating liver cancer. Background technique [0002] Cephalanthine (CEP) is a monomeric compound extracted from Stephania chinensis, a plant of the family Fangjiceae. It belongs to the class of alkaloids. It has been approved by the Japanese Ministry of Health for the clinical treatment of leukopenia and alopecia areata caused by radiotherapy. Wait. Previous studies have shown that stephine has antiviral, anti-inflammatory, anti-tumor and other activities, and no safety problems have been found, and there are few reports of side effects. Stephine can also regulate immunity, maintain cell membrane stability, and promote vascular circulation under reasonable medication. Generally, Stephanine will be prepared in the form of powder, tablet, and injection for treatm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/06A61K31/4741A61K31/4706A61P1/16A61P35/00
CPCA61K31/4706A61K31/4741A61K45/06A61P1/16A61P35/00A61K2300/00
Inventor 王一飞王瑶苏桂锋黄泽秀
Owner GUANGZHOU JINAN BIOMEDICINE RES & DEV CENT
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