Inula wissmannian extract and preparation thereof and application in preparation of antitumor drug

Abstract

The invention provides an inula wissmannian extract and preparation thereof and application in preparation of an antitumor drug. The extract is gemma alkane type sesquiterpene lactone inula wissmannian lactone methyl, ethyl, propyl, butyl and amyl or sheepear inula herb lactone. The extract has the following chemical structural formula and absolute configuration that: due to the toxicity killing function experiment of inula wissmannian lactone methyl, ethyl, propyl, butyl and amyl or sheepear inula herb lactone on human hepatoma cells HepG2, human prostatic cancer cells PC-3, human gastric carcinoma cells MGC-803, human leukemia cells K562, human nasopharynx cancer cells KB and normal human hepatocytes LO2, the result shows that the gemma alkane type sesquiterpene lactone has strong tumor cell killing function, and proliferation of tumor cells can be inhibited, so that the tumor cells are killed. The animal in-vivo experiment proves that the inula wissmannian plant extract and monomeric compounds have the anti-tumor effect and can serve as active ingredients for preparing the antitumor drug. A novel cancer treatment medicine is provided clinically, and high clinical application values and social benefits are generated.

Description

technical field

[0001] The present invention relates to natural medicines, in particular to the extraction and separation method and application of the effective components of S. yunnanensis; in particular, to the extraction and separation method of gemane-type sesquiterpene lactones from S. yunnanensis extract and the preparation of antitumor drugs in the application. Background technique

[0002] Malignant tumors have become a global public health problem. In the past 30 years, the incidence of malignant tumors has increased at an average annual rate of 3%-5%, and 3 / 4 of the new cases occurred in newly industrialized countries and developing countries. Malignant tumors have become the number one cause of death for human beings. Currently in China, malignant tumors have become one of the leading causes of death. With the rapid changes in the economy, society, environment and lifestyle, and the rapid transformation of the population structure towards aging, the cancer spec...

Examples

Embodiment 1

[0043] The preparation of embodiment 1 southern yunnan fennelide A, B, C, D, E and squirrel

[0044] After drying and pulverizing the whole herb of South Yunnan sheep ear chrysanthemum (10.0kg), it was cold soaked and extracted with 95% ethanol (30L) at room temperature 25°C for 4 times (24, 12, 12, 12h), the solvent was recovered, and concentrated to dryness without water soaking. cream (631.4g); the total extract was suspended in 1.5L water and extracted 3 times with 5L petroleum ether, dichloromethane and ethyl acetate successively, and the solvent was recovered under reduced pressure to obtain petroleum ether fraction (133.2g), dichloromethane Methane fractions (179.6 g) and ethyl acetate fractions (28.1 g).

[0045] Put 120 g of the dichloromethane part of the southern yunnan chrysanthemum on a normal-phase silica gel column, and use dichloromethane-methanol (100% dichloromethane, 100:1, 50:1, 20:1, 10:1, 5: 1 and 100% methanol) gradient elution, the eluent was detected ...

Embodiment 2

[0064] Example 2 Killing effect of syringolactone A, B, C, D, E and syringolactone (prepared in Example 1) from southern Yunnan on 5 kinds of human tumor cells

[0065] Effects of Diannan fennelide A, B, C, D, E and fennelide on human liver cancer cell HepG2, human gastric cancer cell MGC-803, human prostate cancer cell PC-3, human leukemia cell K562, human nose Pharyngeal carcinoma cell KB (purchased from the Cell Resource Center of Shanghai Academy of Biological Sciences) was studied.

[0066] The experimental drugs Diannan fennelide A, B, C, D, E and fennelide (prepared in Example 1) were co-incubated with tumor cells for 72 hours; the concentrations of the drugs were set to 0, 0.5, 5, 20 , 50 μM; drugs were dissolved in DMSO (dimethyl sulfoxide). Doxorubicin was used as a positive control, and DMSO was used as a negative control.

[0067] Experimental method: CCK-8 method (cell counting kit-8). Add concentration of 4~6×10 to each well of 96-well plate 4 100uL of cell s...

Embodiment 3

[0072] Example 3 Effects of southern Dianan fennelide A, B, C, D, E and saffrolactone (prepared in Example 1) on the proliferation of human liver cancer cell HepG2 and normal human liver cell LO2

[0073] Diannan fennelide A, B, C, D, E and saffrolactone have selective toxic and killing effects on human liver cancer cells (compared to normal human liver cell lines). Effects on the proliferation of human hepatoma cells HepG2 and normal human hepatocytes LO2.

[0074] Experimental time gradient: 0, 24, 48, 72 hours; concentration gradient setting of experimental drug Diannan fennelide A, B, C, D, E and fennelide: 0, 1, 5, 10, 25 , 50 μM; drug was dissolved in DMSO.

[0075] Experimental method: CCK-8 method. Add concentration of 4~6×10 to each well of 96-well plate 4 100uL of cell suspension per mL, placed in a 37°C, 5% CO2 incubator. After 24 hours, add 1uL / well of different concentrations of drugs, set up duplicate wells, and culture at 37°C and 5% CO2.

[0076] Evaluatio...

technical field

[0001] The present invention relates to natural medicines, in particular to the extraction and separation method and application of the effective components of S. yunnanensis; in particular, to the extraction and separation method of gemane-type sesquiterpene lactones from S. yunnanensis extract and the preparation of antitumor drugs in the application. Background technique

[0002] Malignant tumors have become a global public health problem. In the past 30 years, the incidence of malignant tumors has increased at an average annual rate of 3%-5%, and 3 / 4 of the new cases occurred in newly industrialized countries and developing countries. Malignant tumors have become the number one cause of death for human beings. Currently in China, malignant tumors have become one of the leading causes of death. With the rapid changes in the economy, society, environment and lifestyle, and the rapid transformation of the population structure towards aging, the cancer spec...

Examples

Embodiment 1

[0043] The preparation of embodiment 1 southern yunnan fennelide A, B, C, D, E and squirrel

[0044] After drying and pulverizing the whole herb of South Yunnan sheep ear chrysanthemum (10.0kg), it was cold soaked and extracted with 95% ethanol (30L) at room temperature 25°C for 4 times (24, 12, 12, 12h), the solvent was recovered, and concentrated to dryness without water soaking. cream (631.4g); the total extract was suspended in 1.5L water and extracted 3 times with 5L petroleum ether, dichloromethane and ethyl acetate successively, and the solvent was recovered under reduced pressure to obtain petroleum ether fraction (133.2g), dichloromethane Methane fractions (179.6 g) and ethyl acetate fractions (28.1 g).

[0045] Put 120 g of the dichloromethane part of the southern yunnan chrysanthemum on a normal-phase silica gel column, and use dichloromethane-methanol (100% dichloromethane, 100:1, 50:1, 20:1, 10:1, 5: 1 and 100% methanol) gradient elution, the eluent was detected ...

Embodiment 2

[0064] Example 2 Killing effect of syringolactone A, B, C, D, E and syringolactone (prepared in Example 1) from southern Yunnan on 5 kinds of human tumor cells

[0065] Effects of Diannan fennelide A, B, C, D, E and fennelide on human liver cancer cell HepG2, human gastric cancer cell MGC-803, human prostate cancer cell PC-3, human leukemia cell K562, human nose Pharyngeal carcinoma cell KB (purchased from the Cell Resource Center of Shanghai Academy of Biological Sciences) was studied.

[0066] The experimental drugs Diannan fennelide A, B, C, D, E and fennelide (prepared in Example 1) were co-incubated with tumor cells for 72 hours; the concentrations of the drugs were set to 0, 0.5, 5, 20 , 50 μM; drugs were dissolved in DMSO (dimethyl sulfoxide). Doxorubicin was used as a positive control, and DMSO was used as a negative control.

[0067] Experimental method: CCK-8 method (cell counting kit-8). Add concentration of 4~6×10 to each well of 96-well plate 4 100uL of cell s...

Embodiment 3

[0072] Example 3 Effects of southern Dianan fennelide A, B, C, D, E and saffrolactone (prepared in Example 1) on the proliferation of human liver cancer cell HepG2 and normal human liver cell LO2

[0073] Diannan fennelide A, B, C, D, E and saffrolactone have selective toxic and killing effects on human liver cancer cells (compared to normal human liver cell lines). Effects on the proliferation of human hepatoma cells HepG2 and normal human hepatocytes LO2.

[0074] Experimental time gradient: 0, 24, 48, 72 hours; concentration gradient setting of experimental drug Diannan fennelide A, B, C, D, E and fennelide: 0, 1, 5, 10, 25 , 50 μM; drug was dissolved in DMSO.

[0075] Experimental method: CCK-8 method. Add concentration of 4~6×10 to each well of 96-well plate 4 100uL of cell suspension per mL, placed in a 37°C, 5% CO2 incubator. After 24 hours, add 1uL / well of different concentrations of drugs, set up duplicate wells, and culture at 37°C and 5% CO2.

[0076] Evaluatio...

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