Two alkaloid compounds in narcissus roots as well as preparation method and application thereof

Abstract

The invention discloses two alkaloid compounds in narcissus roots as well as a preparation method and application thereof. According to the invention, two alkaloid compounds with unreported structures are separated from narcissus roots. Activity studies show that the two alkaloid compounds with novel structures can effectively inhibit NO release in RAW264.7 cells. As an anti-inflammatory model recognized in the field, an activity research result proves that the alkaloid compound separated from the narcissus root can be used for developing and preparing an anti-inflammatory drug.

Description

Technical Field

[0001] The invention belongs to the field of natural drug chemistry and relates to the discovery, preparation and application of new natural products, and specifically to two alkaloid compounds in narcissus roots and preparation methods and uses thereof. Background Art

[0002] Narcissus tazetta L.var.chinensis Roem. is the dried bulb of the Narcissus plant of the Amaryllis family. It is a perennial herb with an ovoid bulb and is mainly distributed in Jiangsu, Zhejiang, Fujian, Sichuan, Guizhou and other places in my country.

[0003] Pharmacopoeia records show that narcissus root has the effects of clearing away heat and detoxifying, dispersing nodules and reducing swelling. It can be used externally to treat sores and carbuncles, ulcers, mumps, parotitis and insect bites.

[0004] During the research on the chemical components of narcissus root, a new natural product was discovered, and the present invention is specially proposed. Summary of the invention

[0005] The first purpose of the present invention is to provide two alkaloid compounds in the root of narcissus, the second purpose is to provide a preparation method of the alkaloid compounds, and the third purpose is to provide uses of the alkaloid compounds.

[0006] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0007] An alkaloid compound having a structural formula as shown in Formula I or II, or a pharmaceutically acceptable salt or solvate thereof;

[0008]

[0009] A method for preparing the alkaloid compound represented by the above formula I comprises the steps of:

[0010] (1) extracting the root of narcissus root by heating with 95% by volume ethanol aqueous solution, filtering the extract, and concentrating until there is no alcohol taste to obtain an extract concentrate; loading the extract concentrate on a D-101 macroporous resin column, eluting with water, 30% by volume methanol aqueous solution, 60% by volume methanol aqueous solution, and pure methanol in sequence, combining the eluates of different solvents, and concentrating under reduced pressure to obtain extracts of different concentrations of eluted parts;

[0011] (2) taking the extract of the 60% methanol aqueous solution elution portion obtained in step (1), loading it onto a silica gel column, performing column chromatography separation, and then eluting it with a dichloromethane-methanol solvent gradient with a volume ratio of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, and 0:100 in sequence, and analyzing and combining them by thin layer chromatography to obtain 6 fractions A, B, C, D, E, and F;

[0012] (3) The fraction C obtained in step (2) was dissolved in methanol, separated by preparative liquid chromatography, and isocratically eluted with acetonitrile-water in a volume ratio of 18:82. The eluate corresponding to the chromatographic peak with a retention time of 57 min was collected, and concentrated and dried to obtain the obtained product. The chromatographic conditions of the preparative liquid chromatography were: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min.

[0013] Preferably, the solvent for heated extraction in step (1) is an ethanol aqueous solution with a volume fraction of 95%.

[0014] Preferably, in step (2), dry loading is adopted for silica gel column chromatography separation: the extract is dissolved in methanol, the sample is dry mixed with silica gel, and the sample is loaded onto the silica gel column.

[0015] A method for preparing the alkaloid compound represented by the above formula II comprises the steps of:

[0016] (1) extracting the root of narcissus root by heating with 95% by volume ethanol aqueous solution, filtering the extract, and concentrating until there is no alcohol taste to obtain an extract concentrate; loading the extract concentrate on a D-101 macroporous resin column, eluting with water, 30% by volume methanol aqueous solution, 60% by volume methanol aqueous solution, and pure methanol in sequence, combining the eluates of different solvents, and concentrating under reduced pressure to obtain extracts of different concentrations of eluted parts;

[0017] (2) taking the extract of the 60% methanol aqueous solution elution portion obtained in step (1), loading it onto a silica gel column, performing column chromatography separation, and then eluting it with a dichloromethane-methanol solvent gradient with a volume ratio of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, and 0:100 in sequence, and analyzing and combining them by thin layer chromatography to obtain 6 fractions A, B, C, D, E, and F;

[0018] (3) The fraction C obtained in step (2) was dissolved in methanol, separated by preparative liquid chromatography, and isocratically eluted with acetonitrile-water in a volume ratio of 18:82. The eluate at 32-33 min was collected, concentrated and dried to obtain subfraction C-1; the eluate at 40-41 min was collected, concentrated and dried to obtain subfraction C-2; the chromatographic conditions of the preparative liquid chromatography were: YMC-Triart C 18Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min;

[0019] (4) Dissolve the sub-fraction C-2 obtained in step (3) in methanol, separate by preparative liquid chromatography, use acetonitrile-water with a volume ratio of 10:90 as the mobile phase for elution, collect the eluate corresponding to the chromatographic peak with a retention time of 19 min, and concentrate and dry to obtain the eluate; the chromatographic conditions of the preparative liquid chromatography are: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent 4mL / min.

[0020] Preferably, the solvent for heated extraction in step (1) is an ethanol aqueous solution with a volume fraction of 95%.

[0021] Preferably, in step (2), dry loading is adopted for silica gel column chromatography separation: the extract is dissolved in methanol, the sample is dry mixed with silica gel, and the sample is loaded onto the silica gel column.

[0022] The alkaloid compound represented by the above formula I or II or its pharmaceutically acceptable salt or solvate is used for preparing anti-inflammatory drugs.

[0023] Preferably, the anti-inflammatory drug uses the alkaloid compound represented by Formula I or II or its pharmaceutically acceptable salt or solvate as an active ingredient, and is prepared into a pharmaceutically acceptable dosage form through a pharmaceutically acceptable carrier or excipient.

[0024] Preferably, the dosage forms include tablets, capsules, injections, aerosols, pills and pastes.

[0025] Beneficial effects:

[0026] The present invention separates two alkaloid compounds whose structures have not been reported from the root of narcissus. Activity studies show that these two novel alkaloid compounds can effectively inhibit the release of NO in RAW264.7 cells. As an anti-inflammatory model recognized in the art, the activity study results prove that the alkaloid compounds separated from the root of narcissus in the present invention can be used to develop and prepare anti-inflammatory drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The chemical structural formulas of compounds 1 and 2 prepared in Example 1;

[0028] Figure 2 , 3 is the mass spectrum of compounds 1 and 2;

[0029] Figure 4 , 5 For compounds 1 and 2 1 HNMR spectrum;

[0030] Figure 6 , 7 For compounds 1 and 2 13 C NMR spectrum;

[0031] Figure 8 , 9 HSQC spectra of compounds 1 and 2;

[0032] Fig.10 , 11 is the HMBC spectrum of compounds 1 and 2;

[0033] Fig.12 , 13 NOESY spectra of compounds 1 and 2;

[0034] Fig.14 , 15 For compounds 1 and 2 1 H- 1 H COSY spectrum. DETAILED DESCRIPTION

[0035] The essential contents of the present invention are described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0036] Example 1: Preparation and structural confirmation of new compounds

[0037] 1. Preparation method

[0038] This embodiment provides a method for preparing two alkaloid compounds from the root of narcissus, the steps are:

[0039] (1) 10 kg of narcissus root slices were taken, 10 times the volume of 95% ethanol aqueous solution was added at a ratio of 1 kg:10 L, and refluxed for extraction twice at 78° C., each extraction for 2 h, filtered after extraction, combined the extracts, and concentrated until there was no alcohol taste to obtain an extract concentrate; the extract concentrate was loaded on a D-101 macroporous resin column (inner diameter 20 cm×column bed height 100 cm), and eluted with 60 L of water, 60 L of 30% methanol aqueous solution, 60 L of 60% methanol aqueous solution, and 60 L of pure methanol in sequence, and the eluates from different solvents were combined respectively, and concentrated under reduced pressure at 60° C. to obtain extracts of different concentrations of eluted parts;

[0040] (2) Take the extract (29.42 g) of the 60% methanol aqueous solution elution part obtained in step (1), dissolve it with methanol, dry-mix it with silica gel (100-200 mesh, mass is twice the mass of the extract), load it onto a silica gel (100-200 mesh) column (inner diameter 8 cm×column bed height 80 cm), perform column chromatography separation, and then use dichloromethane-methanol solvent gradient elution with volume ratios of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, and 0:100 in sequence, eluting with 20 L of each solvent, and analyzing and combining it by thin layer chromatography (combining the eluates with the same thin layer chromatography development height) to obtain 6 fractions A, B, C, D, E, and F;

[0041] (3) The fraction C obtained in step (2) was dissolved in methanol, separated by preparative liquid chromatography, and isocratically eluted with acetonitrile-water in a volume ratio of 18:82. The eluate corresponding to the chromatographic peak with a retention time of 57 min was collected, concentrated and dried to obtain the target compound 1 (3.1 mg) with a purity of 98%; the eluate at 32-33 min was collected, concentrated and dried to obtain subfraction C-1; the eluate at 40-41 min was collected, concentrated and dried to obtain subfraction C-2; the chromatographic conditions of the preparative liquid chromatography were: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min;

[0042] (4) The sub-fraction C-2 obtained in step (3) was dissolved in methanol and separated by preparative liquid chromatography. Acetonitrile-water with a volume ratio of 10:90 was used as the mobile phase for elution. The eluate corresponding to the chromatographic peak with a retention time of 19 min was collected and concentrated to dryness to obtain the target compound 2 (4.3 mg) with a purity of 98%. The chromatographic conditions of the preparative liquid chromatography were: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent 4mL / min.

[0043] 2. Structural Confirmation

[0044] Structural analysis of compounds: mainly using spectroscopy techniques, including mass spectrometry, nuclear magnetic resonance ( 1 H-NMR, 13 C-NMR, 2D-NMR) to identify its structure, the spectrum is as follows Figure 2 to Figure 15 As shown, the spectral data and analysis process are as follows.

[0045] 1. Structural analysis of target compound 1

[0046] Compound 1 is a yellow powder, high-resolution mass spectrum ESI-TOF-MS: 326.1384 [M+H] + The molecular formula of compound 1 is C19 H 19 NO 4 The exact molecular weight is 326.1384 and the degree of unsaturation is 11.

[0047] Compound 1 1 H-NMR can be observed in δ H A signal peak is given at 2.82 (3H, s), which is presumed to be a nitrogen methyl signal, δ H 3.08 (1H, dd, J=9.8, 1.8Hz, H-6), δ H 3.83 (1H, dd, J = 9.6, 1.8 Hz, H-6) is speculated to be the proton signal of two different chemical environments on the methylene group. H 3.47 (3H, s) is speculated to be a methoxy signal, δ H 4.47 (1H, d, J = 7.8 Hz, H-3) is speculated to be the proton signal on the methine. The aromatic region shows proton signals on 7 aromatic rings, among which δ H 6.64 (1H, d, J=6.0Hz, H-5”), δ H 6.82(1H,d,J=1.8Hz,H-2”),δ H 6.83 (1H, dd, J = 7.8, 1.8 Hz, H-6") is speculated to be the proton signal on the 1,3,4-trisubstituted benzene ring, δ H 6.65(2H, d, J=1.8Hz, H-3', 5'), δ H 6.98 (2H, d, J = 8.4 Hz, H-2', 6') is speculated to be the proton signal of the other disubstituted proton on the benzene ring, δ H 7.20 (1H, d, J = 2.4 Hz, H-4) is inferred to be the olefin proton signal based on the chemical shift value. 13 The C-NMR spectrum shows 19 carbon signals, with a low field area of ​​δ C 169.0 (C-2) is the carbonyl carbon signal, which can be assigned to 2 methyl, 1 methylene, 9 methine, and 6 quaternary carbon signals according to the DEPT spectrum (Table 1). Combining the DEPT, HSQC, and HMBC spectra, it is inferred that δ C 156.5(C-4') and δ C 126.3 (C-1') is a quaternary carbon on a disubstituted benzene ring, δ C 128.2 (C-2', 6') and δ C 116.0 (C-3', 5') is a tertiary carbon on a disubstituted benzene ring. C 147.8(C-3”),δ C 147.7 (C-4") and δ C131.2 (C-1") is the three carbon signals on the 1,3,4-trisubstituted benzene ring, δ C 124.8(C-6”),δ C 115.7 (C-5") and δ C 113.4 (C-2") is the carbon signal on the 1,3,4-trisubstituted benzene ring. 1 H-NMR data, estimated δ C 131.2 (C-4) is a carbon signal in a double bond. C 56.6 (C-6) is the methylene carbon signal at C-6, and it is inferred that δ C 55.6 is the signal of the methoxy carbon on the 1,3,4-trisubstituted benzene ring, and it is inferred that δ C 30.1 is the nitrogen methyl carbon signal. 1 H- 1 In the H COSY spectrum, H-3(δ H 4.47, 1H, d, J = 7.8 Hz) and H-4 (δ H 7.20, 1H, d, J = 2.4 Hz) is directly related (forming a spin coupling system), H-3 (δ H 4.47, 1H, d, J = 7.8 Hz) and H-6 (δ H 3.08, 1H, dd, J=9.8, 1.8Hz, δ H 3.83, 1H, dd, J = 9.6, 1.8 Hz), H-4 (δ H 7.20, 1H, d, J = 2.4 Hz) and H-6 (δ H 3.08, 1H, dd, J = 9.8, 1.8 Hz). Combined with the HMBC spectrum, it can be observed that N-CH 3 Related to C-2 and C-6, indicating that C-2, C-6, N-CH 3 They are connected through nitrogen atoms. H-3 is related to C-2, C-4, C-5 and C-6, H-4 is related to C-2, C-3 and C-5, and H-6 is related to C-2, C-3, C-4 and C-5, indicating that C-2-C-3-C-4-C-5-C-6 is an integral fragment and connected to nitrogen atoms at both ends. In the HMBC spectrum, H-2' / H-6' is related to C-3 and C-4', H-3' / H-5' is related to C-1' and C-4', and H-3 is related to C-1' and C-2' / C-6', indicating that C-1' on the disubstituted benzene ring is connected to C-3. H-2" / H-6" is related to C-5, and H-4 and H-6 are related to C-1", indicating that C-1" on the 1,3,4-trisubstituted benzene ring is connected to C-5. O-CH 3 Related to C-2”, C-3” / C-4”, indicating O-CH3 Connected to C-3". Based on the above data, the planar structure of compound 1 can be determined as Figure 1 As shown in 1, the compound was named 5-(4-hydroxy-3-methoxyphenyl)-3-(4-hydroxyphenyl)-N-methyl-3,6-dihydropyridin-2(1H)-one.

[0048] 1 H NMR (600 MHz, DMSO): δ H 7.20 (1H, d, J = 2.4 Hz, H-4), δ H 6.98(2H,d,J=8.4Hz,H-2',6'), δ H 6.83(1H,dd,J=7.8,1.8Hz,H-6”), δ H 6.82(1H,d,J=1.8Hz,H-2”), δ H 6.65(2H,d,J=1.8Hz,H-3',5'), δ H 6.64(1H,d,J=6.0Hz,H-5”), δ H 4.47 (1H, d, J = 7.8 Hz, H-3), δ H 3.83(1H,dd,J=9.6,1.8Hz,H-6), δ H 3.47(3H,s,O-CH 3 ), δ H 3.08(1H,dd,J=9.8,1.8Hz,H-6), δ H 2.82(3H,s,N-CH 3 ). 13 C NMR (151 MHz, DMSO): δ C 169.0(C-2),156.5(C-4'),147.8(C-3”),147.7(C-4”),133.8(C-5),131.2(C-4),131.2(C-1”),128.2(C- 2',6'),126.3(C-1'),124.9(C-6”),116.0(C-3’,5’),115.7(C-5”),113.4(C-2”),56.6(C-6),55.6(O-CH 3 ), 40.5(C-3), 30.1(N-CH 3 ).

[0049] 2. Structural analysis of target compound 2

[0050] Compound 2 is a yellow powder, high resolution mass spectrum ESI-TOF-MS: 296.1282 [M+H]+ The molecular formula of compound 2 is C 18 H 17 NO 3 The exact molecular weight is 296.1282 and the degree of unsaturation is 11.

[0051] Compound 2 1 H-NMR, 13 The C-NMR spectrum (Table 1) is very similar to the data of compound 1, except that compound 2 lacks a methoxy signal. By combining DEPT, HMQC, and HMBC, 1 H- 1 HCOSY spectrum confirmed the structure of the compound as Figure 1 As shown in Figure 2, it is named 3,5-bis(hydroxyphenyl)-N-methyl-3,6-dihydropyridin-2(1H)-one.

[0052] 1 H NMR (600 MHz, DMSO): δ H 7.17 (1H, d, J = 2.4 Hz, H-4), δ H 6.95(2H,d,J=8.4Hz,H-2',6'), δ H 7.18(1H,d,J=8.4Hz,H-6”), δ H 7.18(1H,d,J=8.4Hz,H-2”), δ H 6.63(2H,d,J=8.4Hz,H-3',3”), δ H 6.61(2H,d,J=8.4Hz,H-5',5”), δ H 4.41 (1H, d, J = 7.8 Hz, H-3), δ H 3.81(1H,dd,J=9.6,1.2Hz,H-6), δ H 3.09(1H,dd,J=10.2,1.8Hz,H-6), δ H 2.81(3H,s,N-CH 3 ). 13 CNMR (151MHz, DMSO): δ C 169.0(C-2),156.4(C-4'),115.9(C-3”),158.5(C-4”),133.7(C-5),130.7(C-4),131.5(C-1”),128.2(C-2’ ,6'),125.8(C-1'),132.1(C-2”,6”),115.9(C-3’,3”),115.7(C-5’,5”),56.7(C-6),40.5(C-3),30.1(N-CH3 ).

[0053] Example 2: Activity of new compounds

[0054] 1. Experimental Materials and Instruments

[0055] Compounds 1 and 2 prepared in Example 1; mouse mononuclear macrophage RAW264.7 (Shanghai Cell Bank, Chinese Academy of Sciences), DMEM complete medium (Beijing Solebold Technology Co., Ltd.), fetal bovine serum (FBS) (Beijing Solebold Technology Co., Ltd.), lipopolysaccharide (LPS) (Beijing Solebold Technology Co., Ltd.), DMSO (Shanghai Yisheng Biotechnology Co., Ltd.), NO detection kit (Shanghai Biyuntian Biotechnology Co., Ltd.), phosphate buffered saline (PBS) (Beijing Solebold Technology Co., Ltd.), microplate reader (Molecular Devices: SpectraMax i3, USA), MCO-20ACI CO 2 Constant temperature incubator (Thermo Corporation, USA), BS125S 1 / 10,000 balance (Mettler-Toledo, Switzerland).

[0056] 2. Experimental Methods

[0057] The anti-inflammatory activity of compounds 1 and 2 was evaluated by NO inhibition rate in the commonly used RAW264.7 cells. The specific evaluation method is as follows:

[0058] The compound was dissolved in dimethyl sulfoxide (DMSO) to a 100 mM stock solution and stored in a -20°C refrigerator. When used, it was adjusted to an appropriate concentration using DMEM medium containing 1% fetal bovine serum (FBS). In a clean bench, 5 mg of lipopolysaccharide was dissolved in 5 mL of PBS and dispensed into sterile centrifuge tubes. Each tube of 1 mL was sealed with a sealing film and stored in a -20°C refrigerator.

[0059] RAW264.7 cells in the logarithmic growth phase were taken at a density of 2×10 5 Cells were inoculated at a density of 10 cells / well in a 24-well plate and cultured for 24 hours. The blank group and the model group were replaced with 500 μL DMEM complete medium per well, and the drug-treated group was replaced with 500 μL DMEM complete medium containing different concentrations (3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM) of the compound per well. After 2 hours, 0.5 μL of LPS solution was added to each well except the blank group. After 24 hours of continuous culture, 50 μL of cell supernatant was collected from each well and the wells were repeated 3 times. The absorbance of each well at 540 nm was detected by an enzyme reader. According to the formula [(A 模型 -A 给药 ) / (A 模型 -A 空白)]×100% to calculate the inhibition rate of cellular NO release.

[0060] 3. Experimental Results

[0061] The measurement results are shown in Tables 1 and 2.

[0062] Table 1 Inhibitory effect of different concentrations of compound 1 on LPS-induced NO production in RAW264.7 cells

[0063]

[0064] Table 2 Inhibitory effect of different concentrations of compound 2 on LPS-induced NO production in RAW264.7 cells

[0065]

[0066] The results of the assay showed that as the concentration increased, the NO inhibitory activity of compounds 1 and 2 provided in the examples continued to increase, and showed a certain dose dependence.

[0067] The purpose of the above-mentioned embodiments is to specifically introduce the essential content of the present invention, but those skilled in the art should know that the protection scope of the present invention should not be limited to the specific embodiments.

Claims

1. An alkaloid compound having a structural formula as shown in Formula I or II, or a pharmaceutically acceptable salt or solvate thereof; 2. A method for preparing the alkaloid compound represented by formula I in claim 1, characterized in that: Includes steps: (1) extracting the root of narcissus root by heating with 95% by volume ethanol aqueous solution, filtering the extract, and concentrating until there is no alcohol taste to obtain an extract concentrate; loading the extract concentrate on a D-101 macroporous resin column, eluting with water, 30% by volume methanol aqueous solution, 60% by volume methanol aqueous solution, and pure methanol in sequence, combining the eluates of different solvents, and concentrating under reduced pressure to obtain extracts of different concentrations of eluted parts; (2) taking the extract of the 60% methanol aqueous solution elution portion obtained in step (1), loading it onto a silica gel column, performing column chromatography separation, and then eluting it with a dichloromethane-methanol solvent gradient with a volume ratio of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, and 0:100 in sequence, and analyzing and combining them by thin layer chromatography to obtain 6 fractions A, B, C, D, E, and F; (3) The fraction C obtained in step (2) was dissolved in methanol, separated by preparative liquid chromatography, and isocratically eluted with acetonitrile-water in a volume ratio of 18:

82. The eluate corresponding to the chromatographic peak with a retention time of 57 min was collected, and concentrated and dried to obtain the obtained product. The chromatographic conditions of the preparative liquid chromatography were: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min.

3. The preparation method according to claim 2, characterized in that: The solvent for heating and extraction in step (1) is an ethanol aqueous solution with a volume fraction of 95%.

4. The preparation method according to claim 2, characterized in that: In step (2), dry loading is adopted for separation by silica gel column chromatography: the extract is dissolved in methanol, the sample is dry mixed with silica gel, and the sample is loaded onto the silica gel column.

5. A method for preparing the alkaloid compound represented by formula II in claim 1, characterized in that: Includes steps: (1) extracting the root of narcissus root by heating with 95% by volume ethanol aqueous solution, filtering the extract, and concentrating until there is no alcohol taste to obtain an extract concentrate; loading the extract concentrate on a D-101 macroporous resin column, eluting with water, 30% by volume methanol aqueous solution, 60% by volume methanol aqueous solution, and pure methanol in sequence, combining the eluates of different solvents, and concentrating under reduced pressure to obtain extracts of different concentrations of eluted parts; (2) taking the extract of the 60% methanol aqueous solution elution portion obtained in step (1), loading it onto a silica gel column, performing column chromatography separation, and then eluting it with a dichloromethane-methanol solvent gradient with a volume ratio of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, and 0:100 in sequence, and analyzing and combining them by thin layer chromatography to obtain 6 fractions A, B, C, D, E, and F; (3) Dissolve the fraction C obtained in step (2) in methanol, separate by preparative liquid chromatography, and isocratically elute with acetonitrile-water in a volume ratio of 18:

82. Collect the eluate after 32-33 min, concentrate and dry to obtain subfraction C-1. The eluate at 40-41 min was collected, concentrated and dried to obtain subfraction C-2; the chromatographic conditions of the preparative liquid chromatography were: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min; (4) Dissolve the sub-fraction C-2 obtained in step (3) in methanol, separate by preparative liquid chromatography, use acetonitrile-water with a volume ratio of 10:90 as the mobile phase for elution, collect the eluate corresponding to the chromatographic peak with a retention time of 19 min, and concentrate and dry to obtain the eluate; the chromatographic conditions of the preparative liquid chromatography are: YMC-Triart C 18 Chromatographic column, column size 250mm*10mm, particle size 5μm, eluent 4mL / min.

6. The preparation method according to claim 5, characterized in that: The solvent for heating and extraction in step (1) is an ethanol aqueous solution with a volume fraction of 95%.

7. The preparation method according to claim 5, characterized in that: In step (2), dry loading is adopted for separation by silica gel column chromatography: the extract is dissolved in methanol, the sample is dry mixed with silica gel, and the sample is loaded onto the silica gel column.

8. Use of the alkaloid compound represented by formula I or II or its pharmaceutically acceptable salt or solvate as claimed in claim 1 for preparing anti-inflammatory drugs.

9. The use according to claim 8, characterized in that: The anti-inflammatory drug uses the alkaloid compound represented by formula I or II or its pharmaceutically acceptable salt or solvate as an active ingredient, and is prepared into a pharmaceutically acceptable dosage form through a pharmaceutically acceptable carrier or excipient.

10. The use according to claim 9, characterized in that: The dosage forms include tablets, capsules, injections, aerosols, pills and ointments.

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