A kind of timasteroid saponin compound and its preparation method and application

Abstract

The invention discloses a timasteroid saponin compound, a preparation method thereof and the application of the compound in the preparation of antitumor drugs. The compound is a spirostanol-type steroidal saponin with the molecular formula C 45 h 74 o 17 , the molecular weight is 886. The compound is extracted from the medicinal plant Zhimu by 75% ethanol, water-saturated n-butanol, and separated and purified by macroporous adsorption resin, silica gel column chromatography, ODS reversed-phase column chromatography, gel column chromatography and other methods. . The results of MTT experiments show that the compound has significant anti-tumor activity against human breast cancer cell line (MCF‑7) and human liver cancer cell line (HepG2), and is expected to become an anti-tumor drug candidate.

Description

technical field

[0001] The invention belongs to the technical field of medicine, and in particular relates to a timsteroid saponin compound, a preparation method thereof, and an application in the preparation of antitumor drugs. Background technique

[0002] As the main cause of human death, cancer has become a global public health problem. Because cancer is characterized by high mortality, low treatment efficiency, and difficulty in controlling, many scientists have devoted themselves to developing new treatments and novel chemotherapy strategies for the treatment of various cancers. However, because some treatment options have obvious side effects and high treatment costs, countries have turned their attention to natural medicines in recent years.

[0003] Natural medicines have gradually become the focus of research on cancer therapeutic drugs due to their wide range of targets and low toxicity, and great progress has been made in cancer treatment. At present, many drug...

Examples

Embodiment 1

[0035] Preparation of Steroidal Saponin Compounds

[0036] Weigh 10kg of dried rhizome of Anemarrhena, put it in an extraction tank, add 50L of 75% ethanol, soak overnight, heat and reflux in a water bath for extraction for 3 hours, filter, and repeat the extraction with 75% ethanol (50L) twice, each time for 3 hours, and combine three times The filtrate was concentrated to obtain extract-like ethanol extract (3.09kg). Suspend the ethanol extract in pure water to obtain a suspension, extract three times successively with equal volumes of petroleum ether, ethyl acetate, and water-saturated n-butanol, and concentrate the extract to obtain three parts of the extract. The n-butanol part extract (600g) is roughly divided by macroporous adsorption resin, and the first gradient elution is carried out with 10%, 30%, 50%, 70%, 95% (v / v) ethanol-water, The eluate from the 70% ethanol part was crudely separated by silica gel column chromatography. Then use 20:1-1:1 (v / v) dichloromethan...

Embodiment 2

[0050] Antitumor activity test of compound I

[0051] 1. Cell lines and reagent materials Human breast cancer cell line MCF-7 and human liver cancer cell line HepG2 are provided by the American Type Culture Collection (ATCC); DMEM medium, RPMI-1640 medium, fetal bovine serum, trypsin, DMSO and MTT et al.

[0052] 2. MTT method to detect cell viability

[0053] Cultivate MCF-7 and HepG2 cell lines with RPMI-1640 medium containing 10% fetal bovine serum, respectively take MCF-7 and HepG2 cell lines in logarithmic growth phase and in good condition, discard the medium, and pre-cool aseptic Wash with PBS three times, then digest with trypsin solution and observe with an inverted biological microscope. After the digestion is completed, the cells are mixed evenly. Spread the MCF-7 and HepG2 cell lines on 96-well plates respectively to ensure that the MCF-7 cell line is 2500 cells / well, and the HepG2 cell line is 2000 cells / well, and the drug concentration is set to 100 μmol / L for ...

technical field

[0001] The invention belongs to the technical field of medicine, and in particular relates to a timsteroid saponin compound, a preparation method thereof, and an application in the preparation of antitumor drugs. Background technique

[0002] As the main cause of human death, cancer has become a global public health problem. Because cancer is characterized by high mortality, low treatment efficiency, and difficulty in controlling, many scientists have devoted themselves to developing new treatments and novel chemotherapy strategies for the treatment of various cancers. However, because some treatment options have obvious side effects and high treatment costs, countries have turned their attention to natural medicines in recent years.

[0003] Natural medicines have gradually become the focus of research on cancer therapeutic drugs due to their wide range of targets and low toxicity, and great progress has been made in cancer treatment. At present, many drug...

Examples

Embodiment 1

[0035] Preparation of Steroidal Saponin Compounds

[0036] Weigh 10kg of dried rhizome of Anemarrhena, put it in an extraction tank, add 50L of 75% ethanol, soak overnight, heat and reflux in a water bath for extraction for 3 hours, filter, and repeat the extraction with 75% ethanol (50L) twice, each time for 3 hours, and combine three times The filtrate was concentrated to obtain extract-like ethanol extract (3.09kg). Suspend the ethanol extract in pure water to obtain a suspension, extract three times successively with equal volumes of petroleum ether, ethyl acetate, and water-saturated n-butanol, and concentrate the extract to obtain three parts of the extract. The n-butanol part extract (600g) is roughly divided by macroporous adsorption resin, and the first gradient elution is carried out with 10%, 30%, 50%, 70%, 95% (v / v) ethanol-water, The eluate from the 70% ethanol part was crudely separated by silica gel column chromatography. Then use 20:1-1:1 (v / v) dichloromethan...

Embodiment 2

[0050] Antitumor activity test of compound I

[0051] 1. Cell lines and reagent materials Human breast cancer cell line MCF-7 and human liver cancer cell line HepG2 are provided by the American Type Culture Collection (ATCC); DMEM medium, RPMI-1640 medium, fetal bovine serum, trypsin, DMSO and MTT et al.

[0052] 2. MTT method to detect cell viability

[0053] Cultivate MCF-7 and HepG2 cell lines with RPMI-1640 medium containing 10% fetal bovine serum, respectively take MCF-7 and HepG2 cell lines in logarithmic growth phase and in good condition, discard the medium, and pre-cool aseptic Wash with PBS three times, then digest with trypsin solution and observe with an inverted biological microscope. After the digestion is completed, the cells are mixed evenly. Spread the MCF-7 and HepG2 cell lines on 96-well plates respectively to ensure that the MCF-7 cell line is 2500 cells / well, and the HepG2 cell line is 2000 cells / well, and the drug concentration is set to 100 μmol / L for ...

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