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Application of withaferin A in preparation of antitumor drugs

An anti-tumor drug, ashwagandha technology, applied in the field of biomedicine, can solve the problem of not significantly improving the survival rate of lung cancer, and achieve the effect of inhibiting the migration ability of NSCLC cells, promoting the apoptosis of NSCLC cells, and strong synergistic effect

Pending Publication Date: 2022-02-11
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, the curative effect of traditional cytotoxic drugs has reached a plateau, even with a variety of comprehensive treatment methods, it still cannot significantly improve the survival rate of lung cancer

Method used

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  • Application of withaferin A in preparation of antitumor drugs
  • Application of withaferin A in preparation of antitumor drugs
  • Application of withaferin A in preparation of antitumor drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] In this example, the anti-proliferation effect of WA on three types of lung cancer cells (H460, H1975 and PC-9) was detected by MTT assay. The specific process is as follows:

[0024] Spread tumor cells in a 96-well plate at a density of 3000-5000 / well, and place them in a constant temperature incubator at 37°C overnight to allow them to adhere to the wall. The next day, 100 μL of fresh complete medium was replaced, and 1 μL / well of corresponding drug concentration (0.1-20 μM) was added. After 48 hours, 25 μL / well of MTT working solution (5 mg / ml) was added in a dark environment. Discard the supernatant after 3-4 hours, add 150 μL / well of DMSO, shake for 5 minutes to fully dissolve, place in a microplate reader, select the detection wavelength at 490 nm, and measure the absorbance (OD) of each well. The result is as figure 1 As shown in B, the half maximal inhibitory concentrations (Half maximalinhibitory concentration, IC50) of WA in H460, H1975 and PC-9 cells were ...

Embodiment 2

[0028] This example explores the effect of WA on the apoptosis of NSCLC cell lines. After the cells were treated with gradient concentrations of WA, the cells were double-labeled with Annexin V-FITC and PI. The results showed that WA treatment could promote the apoptosis of H460, PC-9 and H1975 cells ( figure 2 A). Using Hoechest combined with live cell DNA to distinguish normal cells from mid-to-early apoptotic cells, the results showed that the proportion of mid-to-early apoptotic cells increased after WA treatment. Further detection of apoptotic proteins suggested that WA treatment could induce the expression of apoptotic proteins Cl-PARP and Bax, while inhibiting the expression of anti-apoptotic protein Bcl-2. These results suggest that WA can promote apoptosis in NSCLC cells.

Embodiment 3

[0030] Transwell migration assay was used to detect the effect of WA on cell crawling ability. Add 500 μL / well basal medium to a 24-well plate and put it into a small chamber with a polycarbonate filter membrane. After the cells were digested and centrifuged, the supernatant was discarded, and the cells were resuspended with serum-free basal medium, counted and diluted, and added to the chamber, about 8000 cells / well. After 24 hours, add 500 μL / well fresh complete medium to a new 24-well plate, transfer the small chamber to a new plate, replace 100 μL serum-free basal medium in the small chamber, and then add 1 μL / well of the corresponding concentration of the drug ( 0, 0.25, 0.5, 0.75mM). After 12-24 hours, fix with 4% paraformaldehyde for 15 minutes, and stain with 1% crystal violet for 15 minutes. Gently wipe off the non-migrated cells inside the chamber with a cotton swab, observe and count the number of migrated cells under an optical microscope. The results showed tha...

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Abstract

The invention discloses an application of withaferin A in preparation of an anti-tumor drug. The anti-tumor drug is used for treating lung cancer. A test result shows that the withaferin A can inhibit NSCLC cell proliferation, promote NSCLC cell apoptosis and inhibit NSCLC cell migration ability; and meanwhile, the combination of the withaferin A and erlotinib or the osimertinib can play a relatively strong synergistic effect, so that the effects of inhibiting NSCLC proliferation and promoting NSCLC apoptosis are achieved.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of withaferin A in the preparation of antitumor drugs. Background technique [0002] Lung cancer is one of the most common malignant tumors in clinical practice, and it is extremely harmful to human health. Related reports show that in recent years, the incidence of lung cancer in various countries has increased by an average of 15% to 65%, and it has become one of the cancers with the highest morbidity and mortality in the world. The 5-year overall survival rate ranges from 4% to 17%, still lower than most other types of cancer. Among all lung cancers, non-small cell lung cancer (NSCLC) accounts for 75% to 80%, and is the most common clinical histological type of lung cancer. Since the early stage disease is usually asymptomatic, most patients (61%) are diagnosed as stage III or stage IV, at which time the chance of surgery has been lost, and chemotherapy...

Claims

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Application Information

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IPC IPC(8): A61K31/585A61K31/517A61K31/506A61P35/00
CPCA61K31/585A61K31/517A61K31/506A61P35/00A61K2300/00
Inventor 黄晓颖赵承光杨乐和郑丹丹周峰朱文静王岩茂王丹丹姚丹王良兴
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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