Stilbene derivative and application thereof

Abstract

The invention provides a stilbene derivative which is prepared by the following steps: crushing caragana angustifolia herb, adding an ethanol solution for reflux extraction, carrying out vacuum concentration and drying on an extracting solution to obtain an extract, dispersing with water to obtain an aqueous dispersion of the extract, and sequentially extracting with petroleum ether, chloroform and ethyl acetate which are equal in volume to obtain the stilbene derivative. And recovering the obtained ethyl acetate extract to obtain an ethyl acetate extract, eluting with silica gel column chromatography, carrying out gradient elution with chloroform and methanol as eluents to obtain six fractions, collecting an eluent part with a volume ratio of 5: 1, carrying out MCI column chromatography, eluting with a 40-70% methanol aqueous solution, collecting a 50% methanol aqueous solution elution part, carrying out high performance liquid chromatography, collecting a 15.7 min peak, and recovering the solvent to obtain the product. The stilbene derivative is obtained. The invention further provides application, and the stilbene derivative can remarkably inhibit proliferation of tumor cells HepG2, Hela and A549 and can be used for preparing antitumor drugs and drugs for preventing tumors.

Description

Technical Field

[0001] The present invention belongs to the technical field of stilbene derivatives, and particularly relates to a stilbene derivative and application thereof. Background Art

[0002] Caragana angustifolia Caragana stenophylla Pojark. is a plant of the genus Caragana in the Leguminosae family, distributed in dry hillsides, piedmont plains, river valleys, and sandy areas of northern Xinjiang, my country. Caragana angustifolia has benefits such as promoting blood circulation, dispelling wind, reducing swelling, and acting as a diuretic. However, due to its unique growth environment, little research has been conducted on its chemical composition, primarily flavonoids and lignans. Studies on its physiological functions are also limited, particularly regarding the potential effects of its astragalus components on tumors.

[0003] The prevention and treatment of tumors is a major challenge facing modern medicine. Stilbene derivatives from Caragana angustifolia have demonstrated promising activity against cervical, lung, and liver cancers, with minimal adverse reactions, low cost, and widespread availability, attracting considerable attention from researchers both in China and abroad. Caragana angustifolia also contains a rich supply of stilbene compounds, but their research, development, and utilization remain underresearched. Therefore, developing and utilizing stilbene derivatives from Caragana angustifolia, determining and characterizing the structures and physicochemical properties of these monomeric compounds, and further exploring their potential medicinal value are of great significance for the development and utilization of Caragana angustifolia. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a stilbene derivative and its application in view of the above-mentioned deficiencies in the prior art. The stilbene derivative can significantly inhibit the proliferation of tumor cells HepG2, Hela and A549, and is used to prepare anti-tumor and tumor prevention drugs.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is: a stilbene derivative, the structural formula of the stilbene derivative is: .

[0006] The present invention also provides a method for preparing the above-mentioned stilbene derivatives, which comprises: S1, Caragana angustifolia Caragana stenophylla Pojark. The whole plant is crushed to obtain anterior leaf Caragana crushed material; S2. Adding ethanol solution to the crushed material of Caragana angustifolia obtained in S1, and performing reflux extraction to obtain an extract; S3, concentrating the extract obtained in S2 under reduced pressure and drying to obtain an extract; S4, dispersing the extract obtained in S3 with water to obtain an aqueous dispersion of the extract; extracting the aqueous dispersion of the extract with an equal volume of petroleum ether, an equal volume of chloroform, and an equal volume of ethyl acetate, three times each, and recovering the obtained ethyl acetate extract to obtain an ethyl acetate extract; S5. The ethyl acetate extract obtained in S4 was eluted by silica gel column chromatography, and chloroform and methanol were used as eluents for gradient elution to obtain 6 fractions, with the volume ratio of chloroform to methanol being 10:1, 5:1, 4:1, 3:1, 2:1, and 1:1, respectively; the eluate portion with a volume ratio of chloroform to methanol of 5:1 was collected, and eluted with 40% to 70% methanol aqueous solution by MCI column chromatography, and the elution portion with 50% methanol aqueous solution was collected; and then subjected to high performance liquid chromatography, the peak at 15.7 min was collected, and the solvent was recovered to obtain a stilbene derivative; the structural formula of the stilbene derivative is: .

[0007] Preferably, the mass fraction of the ethanol solution in S2 is 95%; the mass ratio of the narrow-leaved Caragana crushed material and the ethanol solution is 1:8; the reflux conditions in S2 are: temperature 80°C, reflux extraction 3 times, each time 2 hours, and merging to obtain the extract.

[0008] Preferably, the conditions for the reduced pressure concentration in S3 are: 50° C., −0.09 MPa; the drying temperature is 60° C., and the drying time is 48 h.

[0009] Preferably, the ratio of the extract to water in S4 is 1 g:3 mL.

[0010] Preferably, the method for recovering the ethyl acetate extract in S4 is: concentration under reduced pressure.

[0011] Preferably, the mass ratio of ethyl acetate extract to silica gel in the silica gel column chromatography in S5 is 1:10; the amount of each eluent in S5 is 5 times the column bed volume.

[0012] Preferably, the chromatographic column in the high performance liquid chromatography in S5 is ODS C 18 The mobile phase was methanol and water in a volume ratio of 56%:44%.

[0013] The present invention also provides the use of the stilbene derivatives prepared by the above preparation method, wherein the stilbene derivatives are used for preparing anti-tumor and tumor prevention drugs.

[0014] Preferably, the stilbene derivatives are used to inhibit the proliferation of tumor cells HepG2, Hela and A549.

[0015] The stilbene derivatives in the present invention may be compounds represented by the following structural formula or physiologically acceptable salts thereof: Wherein, R1 is one of hydrogen, methyl, methoxy, OGlc, and carbonyl; R2 is one of hydrogen, methyl, methoxy, OGlc, and carbonyl; Preferably, R1 and R2 are the same, hydroxy or methoxy.

[0016] Compared with the prior art, the present invention has the following advantages: The stilbene derivatives of the invention can significantly inhibit the proliferation of tumor cells HepG2, Hela and A549, and can be used for preparing anti-tumor drugs and tumor prevention drugs.

[0017] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 It is the stilbene derivative of Example 1 of the present invention 1 H NMR spectrum.

[0019] Figure 2 It is the stilbene derivative of Example 1 of the present invention 13 C NMR spectrum. DETAILED DESCRIPTION

[0020] Example 1 The structural formula of the stilbene derivatives in this embodiment is: .

[0021] The preparation method of the stilbene derivative is as follows: S1, Caragana angustifolia Caragana stenophylla Pojark. 10kg of the whole herb was crushed to obtain anterior leaf Caragana chinensis crushed material; S2. Add 95% by mass ethanol solution to the angustifolia crushed material obtained in S1, perform reflux extraction at 80° C. for 3 times, each time for 2 h, and combine to obtain an extract; the mass ratio of the angustifolia crushed material to the 95% by mass ethanol solution is 1:8; The mass fraction of the ethanol solution in this embodiment can also be 50% to 95%; the number of reflux extractions can also be 2 times; S3, the extract obtained in S2 is concentrated under reduced pressure and dried to obtain an extract; the conditions for the reduced pressure concentration are 50°C, -0.09MPa; the drying temperature is 60°C, and the drying time is 48h; S4. Dispersing the extract obtained in S3 with water in a ratio of 1 g:3 mL to obtain an aqueous dispersion of the extract; extracting the aqueous dispersion with an equal volume of petroleum ether, an equal volume of chloroform, and an equal volume of ethyl acetate, three times each; and recovering the obtained ethyl acetate extract to obtain an ethyl acetate extract; recovering the ethyl acetate extract by: concentrating under reduced pressure; S5. The ethyl acetate extract obtained in S4 was eluted by silica gel column chromatography (200-300 mesh column chromatography silica gel, the mass ratio of ethyl acetate extract to silica gel was 1:10), and chloroform and methanol were used as eluents for gradient elution to obtain 6 fractions, and the volume ratios of chloroform and methanol were 10:1, 5:1, 4:1, 3:1, 2:1, and 1:1, respectively; the amount of each eluent was 5 times the volume of the column bed, and the eluate with a volume ratio of chloroform and methanol of 5:1 was collected, and eluted by MCI column chromatography with 40% to 70% methanol aqueous solution, and the elution portion with 50% methanol aqueous solution was collected; and then subjected to high performance liquid chromatography, the peak at 15.7 min was collected, and the solvent was recovered to obtain the target compound, which was a stilbene derivative; the chromatographic column in the high performance liquid chromatography was ODS C 18 Column, mobile phase = methanol + water (56% v + 44% v) The structural formula of the stilbene derivative is: .

[0022] The stilbene derivative compounds of this embodiment 1 H-NMR, 13 C-NMR spectrum data are shown in Table 1. 1 H NMR spectrum is shown in Figure 1 、 13 C NMR see Figure 2 .

[0023] Table 1 NMR data of compounds (CD3OD) Example 2 This example describes the use of the stilbene derivatives prepared in Example 1 for inhibiting the proliferation of tumor cells HepG2 (liver cancer cells), Hela (cervical cancer cells), and A549 (non-small cell lung cancer cells), and for preparing anti-tumor and tumor prevention drugs.

[0024] (1) Cell culture Tumor cells HepG2, Hela and A549 were cultured in DMEM (1X) medium (gibco) containing 10% fetal bovine serum (FBS, gibco), 100 U / mL penicillin, 100 μ g / mL streptomycin was cultured at 37°C and 5% CO2 saturated humidity.

[0025] (2) Cell proliferation test The MTT colorimetric assay was used to test the effects of the stilbene derivatives of the present invention on the proliferation of tumor cells HepG2, Hela, and A549. Specifically, tumor cells in the logarithmic growth phase were seeded in a 96-well plate, with 5,000 cells per well, and cultured in a cell culture incubator for 24 hours. The cells were removed and the supernatant was discarded. Different concentrations of the stilbene derivatives of the present invention (1, 5, 10, 20, 40, 60, 70, 80, 90, 100, 150, 200, 300, 400, 800, 900, 1500, 2000, 3000, 4000, 8000, 9000, 8000, 9000, 5000, 8000 μ M) 200 μ L, continue to culture for 48 hours. Take out, add 5 mg / mL MTT solution 20 μ L, culture for 4 hours, take the supernatant, add 150 μ The cells were shaken in 1 L of DMSO for 10 minutes. The OD value was measured at 492 nm using a microplate reader, and the cell inhibition rate was calculated. A blank experimental group and a control group were also established. The control group received paclitaxel. The results are shown in Table 2.

[0026] Table 2 Effects of compounds on tumor cell proliferation As shown in Table 2, the new stilbene derivatives have a μ M range can inhibit the proliferation of tumor cells HepG2, Hela and A549, IC 50 16.39 ± 0.88, 12.55 ± 0.64, and 37.43 ± 1.07, respectively μ M, showed good antitumor activity.

[0027] The above experimental results show that the new stilbene derivatives can significantly inhibit the proliferation of tumor cells HepG2, Hela and A549, indicating that the stilbene derivatives prepared by the present invention have good anti-tumor activity and can be used for the preparation of anti-tumor drugs and tumor prevention drugs.

[0028] The stilbene derivatives in the present invention may be compounds represented by the following structural formula or physiologically acceptable salts thereof: Wherein, R1 is one of hydrogen, methyl, methoxy, OGlc, and carbonyl; R2 is one of hydrogen, methyl, methoxy, OGlc, and carbonyl.

[0029] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent variation made to the above embodiment based on the essence of the invention technology shall still fall within the scope of protection of the technical solution of the present invention.

Claims

1. A stilbene derivative, wherein the structural formula of the stilbene derivative is: 。 2. A method for preparing a stilbene derivative according to claim 1, characterized in that: The method is: S1, Caragana angustifolia Caragana stenophylla Pojark. The whole plant is crushed to obtain anterior leaf Caragana crushed material; S2. Adding ethanol solution to the crushed material of Caragana angustifolia obtained in S1, and performing reflux extraction to obtain an extract; S3, concentrating the extract obtained in S2 under reduced pressure and drying to obtain an extract; S4, dispersing the extract obtained in S3 with water to obtain an aqueous dispersion of the extract; extracting the aqueous dispersion of the extract with an equal volume of petroleum ether, an equal volume of chloroform, and an equal volume of ethyl acetate, three times each, and recovering the obtained ethyl acetate extract to obtain an ethyl acetate extract; S5. The ethyl acetate extract obtained in S4 was eluted by silica gel column chromatography, and chloroform and methanol were used as eluents for gradient elution to obtain 6 fractions, with the volume ratio of chloroform to methanol being 10:1, 5:1, 4:1, 3:1, 2:1, and 1:1, respectively; the eluate portion with a volume ratio of chloroform to methanol of 5:1 was collected, and eluted with 40% to 70% methanol aqueous solution by MCI column chromatography, and the elution portion with 50% methanol aqueous solution was collected; and then subjected to high performance liquid chromatography, the peak at 15.7 min was collected, and the solvent was recovered to obtain a stilbene derivative; the structural formula of the stilbene derivative is: 。 3. The method for preparing a stilbene derivative according to claim 2, wherein: The mass fraction of the ethanol solution in S2 is 95%; the mass ratio of the narrow-leaved Caragana crushed material to the ethanol solution is 1:8; the reflux conditions in S2 are: temperature 80°C, reflux extraction 3 times, each time 2 hours, and merging to obtain the extract.

4. The method for preparing a stilbene derivative according to claim 2, wherein: The conditions for the reduced pressure concentration in S3 are: 50° C., −0.09 MPa; the drying temperature is 60° C., and the drying time is 48 h.

5. The method for preparing a stilbene derivative according to claim 2, wherein: The ratio of the extract to water in S4 is 1 g:3 mL.

6. The method for preparing a stilbene derivative according to claim 2, wherein: The recovery method of the ethyl acetate extract in S4 is: concentration under reduced pressure.

7. The method for preparing a stilbene derivative according to claim 2, wherein: The mass ratio of ethyl acetate extract to silica gel in the silica gel column chromatography in S5 is 1:10; the amount of each eluent in S5 is 5 times the column bed volume.

8. The method for preparing a stilbene derivative according to claim 2, wherein: The HPLC column in S5 is ODS C 18 The mobile phase was methanol and water in a volume ratio of 56%:44%.

9. Use of a stilbene derivative prepared by the preparation method according to any one of claims 2 to 8, characterized in that: The stilbene derivatives are used for preparing anti-tumor and tumor-preventing drugs.

10. The use according to claim 9, characterized in that The stilbene derivatives are used for inhibiting the proliferation of tumor cells HepG2, Hela and A549.

Images