CN108619183A - The simple extraction methods of coumarin kind compound and its application in a kind of root of Dahurain angelica - Google Patents

Abstract

A simple extraction method of coumarin compounds in Angelica dahurica, extracting Angelica dahurica with methanol, the extract mainly contains imperatorin, isoimperatorin and other coumarin compounds, and tested the extract of Angelica dahurica, Europe Antagonism of procuredin, isoimperatorin and aneurysin on histamine release from human mast cells KU812 induced by compound 48/80. The results show that the extraction and preparation process is simple and effective, and it is found that the extracts of Angelica dahurica, imperatorin, isoimperatorin and coralidin can effectively antagonize the release of histamine in human mast cells KU812 caused by compound 48/80. The invention provides a new method and idea for developing new antiallergic drugs.

Description

A simple extraction method and application of coumarin compounds in Angelica dahurica

technical field

The invention belongs to the technical field of medicine, and relates to a simple extraction method and application of coumarin compounds in Angelica dahurica.

Background technique

The chemical composition of Angelica dahurica is complex, mainly including coumarins, alkaloids, volatile oil, sitosterol, etc. Pharmacological studies have shown that Angelica dahurica has antipyretic, analgesic and anti-inflammatory effects, and has important effects on cardiovascular, central nervous system, smooth muscle, anti-tumor, and photosensitivity. significant effect. The good curative effect of coumarin has made the research on the extraction method of coumarin has been well developed, but the polarity of the active ingredients in Angelica dahurica is relatively close, so the extraction and preparation of coumarin substances in Angelica dahurica need cumbersome steps.

Drug-like allergic reactions are caused by drugs directly stimulating mast cells or basophils, causing mast cells to degranulate and release β-hexosaminidase, histamine and other active transmitters. Clinically, anaphylactoid reactions may occur when patients take medicine for the first time, which not only brings huge economic burden to patients, but also causes new pain and even life-threatening. Studies have found that human MrgprX2 receptors can mediate anaphylactoid reactions. When MrgprX2 receptors are stimulated by drugs, the calcium ion concentration in mast cells can increase, triggering degranulation reactions, and then leading to anaphylactoid reactions. Clinically used antiallergic drugs mainly include antihistamine drugs, allergic reaction mediator release agents and hormone drugs. Antagonizing the MrgprX2 receptor can stop the hypersensitivity reaction at the source, but the antiallergic drugs that antagonize the MrgprX2 receptor are still blank. Therefore, the research on MrgprX2 receptor antagonists has attracted more and more attention. The purpose of drugs to antagonize the MrgprX2 receptor is mainly To prevent the occurrence of anaphylactoid reactions from the source and fundamentally solve the hazards of anaphylactoid reactions, it is of great significance to develop and utilize MrgprX2 receptor antagonists or lead compounds to develop antiallergic drug candidates.

Contents of the invention

The purpose of the present invention is to provide a simple extraction method and application of coumarin compounds in Angelica dahurica.

In order to achieve the above object, the technical scheme adopted in the present invention is:

A simple method for extracting coumarin compounds from Angelica dahurica. The coarse powder of Angelica dahurica is ultrasonically filtered with methanol to obtain an extract, and the methanol is removed to obtain coumarin compounds.

The further improvement of the present invention is that the power of the ultrasound is 200W, the frequency is 40kHz, and the time is 30 minutes.

The further improvement of the present invention is that the ratio of the coarse powder of Angelica dahurica to methanol is 1g:3mL.

An application of the coumarin compound extracted based on the above method in the preparation of antiallergic medicine.

A further improvement of the present invention is that the antiallergic drug is a drug for antagonizing MrgprX2 receptor.

A further improvement of the present invention is that the antiallergic drug is a drug used to inhibit the release of histamine from human mast cells KU812.

A further improvement of the present invention is that the antiallergic drug is a clinically acceptable pharmaceutical preparation.

A further improvement of the present invention lies in that the pharmaceutical preparation is tablet, capsule, granule or injection.

Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention uses methanol to ultrasonically extract the traditional Chinese medicine Angelica dahurica to obtain coumarin compounds, the extraction method is simple, and it is clearly proposed that the coumarin extracts can resist anaphylaxis. The pharmacological activity of three coumarin compounds, imperatorin, isoimperatorin and coralidin, has been found to have anti-anaphylaxis pharmacological activity, and can be used to prepare drugs with anti-allergic effects. The present invention uses in vitro cell experiments to verify the anti-allergic activity of the coumarin extract obtained by this extraction method and the antiallergic activity of these three coumarin compounds with human mast cell model KU812, and finds that the extract obtained by this extraction method Both the coumarin extract and the three coumarin compounds can effectively antagonize the release of histamine from human mast cells KU812 induced by compound 48/80. It shows that the Angelica dahurica extract and these three furanocoumarin compounds obtained by applying the extraction method can be used to prepare medicines with antiallergic effects.

Description of drawings

Fig. 1 is the HPLC chromatogram of coumarin extract.

Figure 2 shows that coumarin extract inhibits the release of histamine from KU812 caused by compound 48/80, compound 48/80 is used as a positive control drug, and TM buffer is used as a negative control.

Figure 3 shows that imperatorin inhibits the histamine release of KU812 caused by compound 48/80, compound 48/80 is used as a positive control drug, and TM buffer is used as a negative control.

Figure 4 shows that isoimperatorin inhibits the release of histamine from KU812 caused by compound 48/80, compound 48/80 is used as a positive control drug, and TM buffer is used as a negative control.

Figure 5 shows that pectinoid inhibits the release of histamine from KU812 caused by compound 48/80, compound 48/80 is used as a positive control drug, and TM buffer is used as a negative control.

Detailed ways

The present invention will be described below in conjunction with the accompanying drawings and specific embodiments.

The invention extracts the Angelica dahurica with methanol, and the extract mainly contains coumarin compounds such as imperatorin and isoimperatorin. And investigated the antagonism of the extracts of Angelica dahurica, imperatorin, isoimperatorin and coralidin on the release of histamine from human mast cells KU812 induced by compound 48/80. The research results show that the extraction and preparation process is simple and effective, and it is found that the extracts of Angelica dahurica, imperatorin, isoimperatorin and coralidin can effectively antagonize the release of histamine in human mast cells KU812 caused by compound 48/80. The invention provides a new method and idea for developing new antiallergic drugs.

Example 1

1. Experimental method

The decoction slices of Angelica dahurica were crushed into coarse powder, and 50 g of Angelica dahurica coarse powder was weighed, placed in a stoppered Erlenmeyer flask, 150 mL of methanol was added, and ultrasonic extraction was performed for 30 minutes under the condition of ultrasonic power of 200 W and frequency of 40 kHz. The filtrate was filtered, and the solvent (methanol) was evaporated with a rotary evaporator to obtain the coumarin extract. Additionally, methanol can be recycled.

Example 2

1. Experimental method

(1) Histamine release

KU812 cells were seeded in 96-well plates at 30,000 cells/well and cultured overnight. After the 96-well plate was centrifuged at 1500rmp for 5min, the original culture medium was sucked out, and 90ul TM buffer (negative control group), 30ug/ml C48/80 solution of the drug (administration group) or 30ug/ml C48/80 solution (administration group) were added respectively. Positive control group) was incubated in an incubator at 37°C for 30 minutes, then centrifuged at 1500rmp for 5 minutes, and 50ul of each well was pipetted into ep tubes. Add 100ul of deuterated histamine acetonitrile solution, vortex, centrifuge at 12000×g for 10min, take the supernatant, and measure the content of histamine by HPLC-MS-MS.

2. Experimental results

The data are expressed as mean ± standard error (mean ± SEM), and the graphs are drawn using Graphpad Prism 5.0 software.

Fig. 1 is the HPLC chromatogram of coumarin extracts (1 is imperatorin, 2 is pectinoid, 3 is isoimperatorin).

Fig. 2 is the histamine figure (Control is negative control group, C48/80 is positive control group, *P<0.05, **P<0.01, ** *P<0.001, n=4).

Fig. 3 is the histamine graph of imperatorin inhibiting KU812 caused by compound 48/80 (Control is negative control group, C48/80 is positive control group, *P<0.05, **P<0.01, ***P <0.001, n=4).

Figure 4 is the histamine release graph of isoimperatorin inhibiting KU812 caused by compound 48/80 (Control is a negative control group, C48/80 is a positive control group, *P<0.05, **P<0.01, ** *P<0.001, n=4).

Figure 5 is a map of the histamine release of pectinoids inhibiting KU812 caused by compound 48/80 (Control is a negative control group, C48/80 is a positive control group, *P<0.05, **P<0.01, ***P <0.001, n=4).

Show by above-mentioned experimental result, extraction method of the present invention, convenient, fast, coumarin compound extraction rate height, to gained coumarin class extract, and main component imperatorin thereof, isoimperatorin and All the corallins could effectively antagonize the histamine release of KU812 caused by compound 48/80, among which imperatorin and isoimperatorin were the best.

The main components of coumarin compounds, imperatorin, isoimperatorin and acarinin all have the effect of antagonizing the release of histamine from human mast cells KU812 induced by compound 48/80 and antagonizing the release of histamine from human mast cells KU812 induced by compound 48/80. Role of histamine release from mast cells KU812.

It can be seen that the main components of the coumarin compound extracted in the present invention are imperatorin, isoimperatorin and pectinoid, and the coumarin compound can be used in the preparation of antiallergic drugs. The antiallergic drug is a drug for antagonizing MrgprX2 receptors or the antiallergic drug is a drug for inhibiting the release of histamine from human mast cells KU812. Meanwhile, the antiallergic medicine is a clinically acceptable pharmaceutical preparation, which is tablet, capsule, granule or injection.

The present invention discovers a simple extraction method of coumarin compounds in the traditional Chinese medicine Angelica dahuricae, and clearly proposes the obtained coumarin extract, and its main components imperatorin, isoimperatorin and pectinoids. The pharmacological activity of anaphylaxis, which is a new pharmacological effect of coumarin compounds, that is, it has a good antagonism on the release of KU812 caused by compound 48/80, and has opened up a new field of clinical application of traditional Chinese medicine components. It is of great significance to the development of antiallergic drugs.

Claims (8)

1. a simple extraction method of coumarin compounds in Angelica dahurica, it is characterized in that, the Angelica dahurica coarse powder is ultrasonically obtained with methanol, after filtering, extracting solution is obtained, and methanol is removed to obtain coumarin compounds. 2. the simple extraction method of coumarin compound in a kind of Angelica dahurica according to claim 1, is characterized in that, the power of ultrasonic is 200W, and frequency is 40kHz, and the time is 30 minutes. 3. the simple extraction method of the coumarin compound in a kind of Angelica dahurica according to claim 1, is characterized in that, the ratio of the coarse powder of Angelica dahurica and methanol is 1g: 3mL. 4. The application of the coumarin compounds extracted according to any one of the methods of claims 1-3 in the preparation of antiallergic drugs. 5. The antiallergic drug according to claim 4 is a drug for antagonizing the MrgprX2 receptor. 6. The antiallergic drug according to claim 4 is a drug for inhibiting the release of histamine from human mast cells KU812. 7. the antiallergic drug according to claim 4 is a clinically acceptable pharmaceutical preparation. 8. The pharmaceutical preparation according to claim 7 is tablet, capsule, granule or injection.