Holothuria moebii saponin A, and preparation and application thereof

A technology of sea cucumber saponins and sea cucumber total saponins, which is applied in the field of medicine and can solve problems such as severe drug resistance, limited curative effect of glioma drugs, and hindering drug curative effect

Inactive Publication Date: 2014-09-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the curative effect of the existing drugs for treating glioma including temozolomide is limited, and most of them have serious deficiencies. The outstanding performances are: (1) most of the

Method used

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  • Holothuria moebii saponin A, and preparation and application thereof
  • Holothuria moebii saponin A, and preparation and application thereof
  • Holothuria moebii saponin A, and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of sea cucumber saponin A

[0031] (1) Extraction, separation and purification of sea cucumber saponin A

[0032] Cut the frozen sea cucumbers (fifty) into small pieces, and extract them five times by methanol percolation (5000 ml of methanol for the first time, and 3000 ml of methanol each time), the methanol percolation fluids are combined and concentrated under reduced pressure A concentrated methanolic extract was obtained. The concentrated methanol extract was successively extracted with cyclohexane, ethyl acetate and n-butanol, and the n-butanol extracts were combined and concentrated under reduced pressure to obtain n-butanol extract extract (40.0 g). The n-butanol extract extract was dissolved in water, and the water soluble matter was separated by macroporous resin (600 ml) column chromatography, and 10% (v / v) methanol water (4000 ml) and 80% (v / v) Methanol and water (4000 mL) were eluted sequentially. The 80% (v / v) methanol eluates w...

Embodiment 2

[0037] Example 2 The effect of sea cucumber saponin A on inhibiting the proliferation of glioma cells

[0038] Rat glioma C6 cells and human glioma U87-MG, U251 and HSG-44 cells were incubated with DMEM and 10% FBS at 37 and 5% carbon dioxide in an incubator, and the cells that were 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.

[0039] The survival rate of tumor cells was determined by the sulforhodamine B method (SRB). Cells were seeded in 96-well plates, and different concentrations of test drugs were added 24 h after adherence. After 72 h of drug treatment, stain with SRB, measure the absorbance at 515 nm with a microplate reader, detect the survival rate of tumor cells, and calculate the IC 50 value. The results showed that sea cucumber saponin A significantly inhibited the proliferation of glioma...

Embodiment 3

[0041] Example 3 Effects of Mie's sea cucumber saponin A on inducing apoptosis and necrosis of tumor cells

[0042] Double staining with 4',6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) to determine the effect of Mie's sea cucumber saponin A on apoptosis and necrosis in human glioma U87-MG cells . Tumor cells and different concentrations of sea cucumber saponin A at 37 After 72 hours in the incubator, use 10 / mL DAPI and 5 / mL of PI for 20 min 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 prove that: Mie's sea cucumber saponin A (4.0 ) significantly induced apoptosis and necrosis in human glioma U87-MG cells (attached Figure 9 ).

[0043] Using Annexin V-FITC / PI double-staining analysis method...

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Abstract

The invention provides Holothuria moebii saponin A. The Holothuria moebii saponin A is prepared by separating total Holothuria moebii saponin from the marine invertebrate Holothuria moebii through percolation, extraction and reverse phase silica gel (ODS) column chromatography, then carrying out reverse phase silica gel column chromatographic separation and carrying out pressure-reduced concentration. The Holothuria moebii saponin A provided by the invention can substantially inhibit proliferation of a plurality of cells like rat brain glioma C6 and human brain glioma U87-MG, U251 and SHG-44, induce tumor cell apoptosis, reduce protein expression of a plurality of key enzymes in characteristic metabolism process of tumor cells and has a unique multi-target anti-glioma characteristic. Thus, the Holothuria moebii saponin A can be applied in preparation of drugs used for treating brain glioma. The chemical structural formula of the Holothuria moebii saponin A is described in the specification.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to Moebioside A (Moebioside A), and sea cucumber Moebioside A obtained from marine invertebrates (Moebioside A) Holothuria moebii ) in the method for preparing the anti-tumor active compound Moebioside A (Moebioside A) and its application in the preparation of medicaments for treating glioma. Background technique [0002] Glioma (glioma) is the most common and malignant brain tumor in the brain, accounting for about 46% of intracranial tumors and 70% of all malignant brain 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. Combining radiotherapy and temozolomide chemotherapy after surgery is the most common method for the treatment of malignant glioma. For newly diagnosed patients, temozolomide...

Claims

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Application Information

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IPC IPC(8): C07J71/00A61K31/7048A61P35/00
Inventor 张治针虞斯然陈璐连晓媛
Owner ZHEJIANG UNIV
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