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Epoxide sterol composition, and preparation and application thereof

A technology of epoxy sterol and composition, applied in the field of medicine, can solve the problems of affecting the curative effect of drugs, hindering the blood-brain barrier, large toxic and side effects, etc., and achieve the effect of good application prospect.

Inactive Publication Date: 2015-05-27
ZHEJIANG UNIV
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  • Description
  • Claims
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Problems solved by technology

However, most of the existing drugs for the treatment of glioma have deficiencies: (1) These drugs are mostly chemical synthetic products with relatively high toxicity and side effects; (2) The increasing drug resistance of tumor cells seriously affects the efficacy of drugs; (3) The obstruction of the blood-brain barrier restricts the treatment of glioma drugs from reaching the brain and exerting their curative effect more effectively

Method used

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  • Epoxide sterol composition, and preparation and application thereof
  • Epoxide sterol composition, and preparation and application thereof
  • Epoxide sterol composition, and preparation and application thereof

Examples

Experimental program
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Embodiment 1

[0032] The preparation of embodiment 1ESC

[0033] (1) Extraction, separation and purification of ESC

[0034] The frozen green sea anemone (10) was cut into small pieces, extracted three times by percolation with methanol, the first time with 2000 mL of methanol for 24 hours, and the last two times with 1000 mL of methanol for 4 hours. The combined methanol percolates were concentrated under reduced pressure to obtain about 300 mL of methanol extract. The methanol extract was extracted three times with cyclohexane, each time with 300 mL of cyclohexane. The cyclohexane extracts were combined and concentrated under reduced pressure to obtain a cyclohexane extract extract (15.6 g). Cyclohexane extract extract was separated by silica gel column chromatography, respectively with different ratios of cyclohexane / ethyl acetate (4:1,2:1,1:1,1:2), ethyl acetate and methanol 1000mL were eluted sequentially. Collect the eluate of cyclohexane / ethyl acetate (1:2) in sections every 100 ...

Embodiment 2E

[0038] Example 2 ESC and ES-A inhibit the growth of glioma cells

[0039] Rat glioma C6 cells and human glioma U251 cells were cultured with DMEM and 10% FBS medium in an incubator at 37°C and 5% carbon dioxide, and the cells cultured for three generations were used for the experimental research of the present invention. Temozolomide (TMZ), the first-line drug currently used in clinical treatment of glioma, was used as a positive control. ESA is composed of two compounds (1:1 g), ES-A and ES-B, and the total content of the two is 97.8%.

[0040] The survival rate of tumor cells was determined by the sulforhodamine B (SRB) method. Cells were seeded in 96-well plates, and different concentrations of ESC and ES-A were added 24 hours after adherence. After 72 hours of drug treatment, stain with SRB, measure the absorbance value at 515nm with a microplate reader, detect the survival rate of tumor cells, and calculate the IC50 values ​​of ESC and ES-A. The experimental results sh...

Embodiment 3

[0041] Example 3 Effects of ES-A on Inducing Tumor Cell Apoptosis and Necrosis

[0042] Double staining with 4',6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) was used to determine the effect of ES-A on inducing apoptosis and necrosis of human glioma U251 cells. Tumor cells and different concentrations of ES-A were incubated in an incubator at 37°C for 72 hours and then stained with 10 μg / mL DAPI and 5 μg / mL PI for 20 minutes at room temperature. After washing twice with PBS, the apoptosis and necrosis of tumor cells were observed under a 40-fold fluorescence microscope. Apoptotic cells were stained with DAPI and stained in bright blue, and necrotic cells were stained with propidium iodide (PI) in red. The experimental results showed that ES-A40 (μM and 80μM) induced apoptosis and necrosis in human glioma U251 cells (attached Figure 13 ).

[0043] Table 1: Nuclear Magnetic Resonance Spectroscopy (NMR) data of ESC and ES-A (in pyridine-d 5 )

[0044]

[004...

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Abstract

The invention provides an epoxide sterol composition comprising epoxide sterol A and epoxide sterol B, of which the total content is greater than 95%. The epoxide sterol composition is prepared by percolation extraction, concentration and column chromatography isolation purification of a marine animal, namely green sea anemone. Experimental results show that the epoxide sterol composition (ESC) remarkably refrains the growth of rat glioma cells C6 and human glioma cells U251, induces tumor cell apoptosis or necrosis, and can be used for preparing medicines for curing brain gliomas.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to the preparation of an epoxy sterol composition and its application in the preparation of anti-glioma drugs. Specifically, the present invention relates to a method for preparing an epoxysterol composition by extracting and separating from the marine animal green sea anemone Anthopleura midori and the application of the composition in treating glioma. Background technique [0002] Glioma (glioma) is the most common intracranial tumor, accounting for about 46% of intracranial tumors. Statistics from the World Health Organization show that malignant glioma is the second cause of death for tumor patients under the age of 34, and the third cause of death for patients aged 35 to 54, seriously endangering human health and life. Similar to the treatment of other tumors, the treatment of glioma mainly includes surgery, radiotherapy and chemotherapy. Chemotherapy is still one of the main methods fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/58A61P35/00
Inventor 张治针虞斯然叶雪威连晓媛
Owner ZHEJIANG UNIV
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