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Application of conyza blinii total saponins for preparing anti-neoplastic drug

An anti-tumor drug, the technology of gentian grass, which is applied in the direction of anti-tumor drugs, drug combinations, pharmaceutical formulas, etc., can solve problems such as unsatisfactory treatment

Inactive Publication Date: 2011-01-05
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cancer treatment methods based on surgery and radiotherapy can only achieve good curative effect on the early stages of cancer, and are not ideal for the treatment of middle and late stages

Method used

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  • Application of conyza blinii total saponins for preparing anti-neoplastic drug
  • Application of conyza blinii total saponins for preparing anti-neoplastic drug
  • Application of conyza blinii total saponins for preparing anti-neoplastic drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Preparation of total saponins of golden gentian:

[0021] The golden gentian was extracted twice with a 95% aqueous ethanol solution by volume, and then extracted once with an aqueous ethanol solution with a volume percentage of 60%; the extracts were filtered, combined, and concentrated under reduced pressure to no ethanol , Then suspended in water to a concentration of about 30%, followed by extraction with ethyl acetate and n-butanol; the n-butanol extract was concentrated under reduced pressure to obtain crude saponin; the crude saponin was subjected to macroporous resin column chromatography, and then water, volume concentration Gradient elution is 30% ethanol aqueous solution, 70% ethanol aqueous solution with volume concentration and 95% ethanol aqueous solution with volume concentration, and finally the 70% ethanol eluate is concentrated under reduced pressure to obtain golden gentian total saponin.

Embodiment 2

[0023] The inhibitory effect of the total saponins of gentian grass on cervical cancer cells (Hela cells):

[0024] Digest Hela cells with 0.25% trypsin digestion solution 1mL at 37°C for 5min. Observe cytoplasmic retraction and intercellular space enlargement under the microscope, that is, after the cells become rounded, stop the digestion; add 10% FBS medium to neutralize the pancreas Enzyme; cell count, dilute the cells to 2×10 4 Cells / mL, add to a 96-well plate, add 200μl cell suspension per well; place the culture plate in an incubator for 24 hours to allow the cells to adhere to the wall, and add drugs when the cells enter the exponential growth phase; Gentiana saponins, prepared in Example 1 of XB-5) and the control group (cisplatin, DDP) drugs are diluted 10 times in series, and the concentration (XB-5 is calculated with M=1200) is 10 -3 M, 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, and as a blank control, set up 5 replicate wells for each drug concentration, and take them out at...

Embodiment 3

[0028] The inhibitory effect of total saponins of gentian grass on lung cancer cells (SPC-A1).

[0029] Digest the SPC-A1 cells with 0.25% trypsin digestion solution 1mL, 37℃ for 5min, observe the cytoplasmic retraction, intercellular space enlargement, that is, after the cells become rounded under the microscope, stop the digestion; add to the culture medium containing 10% FBS And trypsin; count the cells and dilute the cells to 2×10 4 Cells / mL, add to a 96-well plate, add 200μl cell suspension per well; place the culture plate in an incubator for 24 hours to allow the cells to adhere to the wall, and add drugs when the cells enter the exponential growth phase; Gentiana saponins, XB-5) and the control group (Paclitaxel, Paclitaxel) are diluted by 10 times, the concentration (XB-5 is calculated by M=1200) is 10 -3 M, 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, and as a blank control, set up 5 replicate wells for each drug concentration, and take them out at 24, 48 and 72 hours respectivel...

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Abstract

The invention discloses application of conyza blinii total saponins for preparing an anti-neoplastic drug. An MTT method is used for detecting a growth inhibition ratio of the conyza blinii total saponins for cervical carcinoma Hela cells and lung carcinoma SPC-A1 cells and calculating IC50 (50 percent of Inhibiting Concentration); a result explains that the conyza blinii total saponins has time and concentration dependency for the growth inhibition of the Hela cells and the SPC-A1 cells; an agarose gel electrophoresis method is adopted to detect acted cells of the conyza blinii total saponins in order to display an apoptotic characteristic DNA ladder and indicate that the conyza blinii total saponins has an apoptotic function for the Hela cells and the SPC-A1 cells; and a Western-blot method is adopted to detect the influence on apoptotic key protein caspase-3 content by the conyza blinii total saponins, obtain that after the conyza blinii total saponins acts on the cells, casepase-3 is increased through a semiquantitative analysis and indicate that the growth inhibition of the cells by the conyza blinii total saponins is regulated by an apoptosis mechanism.

Description

Technical field [0001] The present invention relates to the use of total saponin of golden gentian. Background technique [0002] Cancer is one of the three major diseases that endanger human health. In the past two decades, the incidence of lung cancer has increased year by year due to increased air pollution, increased smokers, and the emergence of occupations that have long-term exposure to radioactive materials, asbestos and other chemical carcinogens. Lung cancer has become a male cancer patient. Leading cause of death. Cervical cancer is one of the main diseases that endanger women's lives and reduce their quality of life. Globally, more than 200,000 women die from cervical cancer each year. In developing countries, cervical cancer is a common and frequent gynecological tumor, ranking first. Currently in developing countries, the incidence of cervical cancer is the second highest among female malignancies. Surgery and radiotherapy dominated cancer treatment methods can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/28A61P35/00A61P15/00A61P11/00
Inventor 苏艳芳颜世伦周立军刘培
Owner TIANJIN UNIV
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