Two alkaloid compounds in narcissus roots and preparation method and use thereof

Abstract

The application discloses two alkaloid compounds in narcissus tazetta roots, a preparation method and application thereof. Two alkaloid compounds with novel structures are separated from narcissus tazetta roots. Activity research shows 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, the activity research result proves that the alkaloid compounds separated from narcissus tazetta roots can be used for developing and preparing anti-inflammatory drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of natural medicinal chemistry, and relates to discovery, preparation and application of a new natural product, in particular to two alkaloid compounds in narcissus tazetta L.var.chinensis Roem and a preparation method and application thereof. BACKGROUND

[0002] Narcissus tazetta L.var.chinensis Roem is the dried bulb of the plant Narcissus tazetta L.var.chinensis Roem in the Amaryllidaceae family, is a perennial herb, and the bulb is an ovate sphere, and is mainly distributed in Jiangsu, Zhejiang, Fujian, Sichuan and Guizhou in China.

[0003] The pharmacopoeia records show that narcissus tazetta L.var.chinensis Roem has the effects of clearing heat and resolving toxins, resolving masses and detumescence, and can be used for treating sores and carbuncles, ulcers, parotitis, parotitis and hundred-worm bites and the like.

[0004] In the process of studying the chemical components of narcissus tazetta L.var.chinensis Roem, a new natural product is found, and the present application is proposed. SUMMARY

[0005] The first object of the present application is to provide two alkaloid compounds in narcissus tazetta L.var.chinensis Roem, the second object is to provide a preparation method of the alkaloid compounds, and the third object is to provide the use of the alkaloid compounds.

[0006] The above objects of the present application are achieved by the following technical solutions.

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

[0008]

[0009] A preparation method of the alkaloid compound shown in formula I, comprising the following steps:

[0010] (1) taking narcissus tazetta L.var.chinensis Roem medicinal materials, heating extraction with 95% ethanol aqueous solution, filtering the extract, concentrating to no alcohol taste to obtain an extract concentrate; taking the extract concentrate, loading into a D-101 macroporous resin column, eluting with water, 30% methanol aqueous solution, 60% methanol aqueous solution and pure methanol in sequence, respectively combining the elution liquids of different solvents, and reducing pressure to concentrate to obtain the extract of different concentration elution parts;

[0011] (2) Take the 60% methanol aqueous solution elution part extract obtained in step (1) and load it into a silica gel column for column chromatography separation, then gradient elute with dichloromethane-methanol solvents with volume ratios of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, 0:100 in sequence, and analyze and combine using thin layer chromatography to obtain 6 flow parts A, B, C, D, E, F;

[0012] (3) Take the flow part C obtained in step (2) and dissolve it with methanol, separate it by preparative liquid chromatography, isocratically elute it with acetonitrile-water with a volume ratio of 18:82, collect the eluent corresponding to the chromatographic peak with a retention time of 57 min, and concentrate and dry it to obtain it; the chromatographic conditions of the preparative liquid chromatography are as follows: YMC-Triart C 18 chromatographic column, the chromatographic column size is 250 mm*10 mm, the particle size is 5 μm, and the eluent flow rate is 4 mL / min.

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

[0014] Preferably, in step (2), dry loading is used for silica gel column chromatography separation: the extract is dissolved with methanol, the silica gel is dry loaded, and then loaded into a silica gel column.

[0015] A preparation method of an alkaloid compound shown in the above formula II, comprising the steps of:

[0016] (1) Take narcissus root decoction pieces and extract them with 95% volume fraction of ethanol aqueous solution by heating, filter the extract, concentrate it to be alcohol-free, and obtain an extract concentrate; take the extract concentrate and load it into a D-101 macroporous resin column, elute it with water, 30% volume fraction of methanol aqueous solution, 60% volume fraction of methanol aqueous solution, and pure methanol in sequence, combine the eluents of different solvents respectively, and concentrate them under reduced pressure to obtain extract of different concentration elution parts;

[0017] (2) Take the 60% methanol aqueous solution elution part extract obtained in step (1) and load it into a silica gel column for column chromatography separation, then gradient elute with dichloromethane-methanol solvents with volume ratios of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, 0:100 in sequence, and analyze and combine using thin layer chromatography to obtain 6 flow parts A, B, C, D, E, F;

[0018] (3) Take the flow part C obtained in step (2) and dissolve it with methanol, separate it by preparative liquid chromatography, isocratically elute it with acetonitrile-water with a volume ratio of 18:82, collect the eluent corresponding to the chromatographic peak with a retention time of 57 min, and concentrate and dry it to obtain it; the chromatographic conditions of the preparative liquid chromatography are as follows: YMC-Triart C 18Chromatographic column, chromatographic column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min;

[0019] (4) The sub-fraction C-2 obtained in step (3) is dissolved in methanol, and then separated by preparative liquid chromatography, eluted with acetonitrile-water (volume ratio 10:90) as the mobile phase, and the eluent corresponding to the chromatographic peak with a retention time of 19 min is collected and concentrated to dryness to obtain the product; the chromatographic conditions of the preparative liquid chromatography are as follows: YMC-Triart C 18 Chromatographic column, chromatographic column size 250mm*10mm, particle size 5μm, eluent flow rate 4mL / min.

[0020] Preferably, the solvent for heating extraction in step (1) is 95% volume fraction of aqueous ethanol.

[0021] Preferably, in step (2), dry loading is used for silica gel column chromatography separation: the extract is dissolved in methanol, and the silica gel is dry-loaded and loaded into the silica gel column.

[0022] The use of the alkaloid compounds represented by the above formula I or II or the pharmaceutically acceptable salts or solvates thereof for preparing anti-inflammatory drugs.

[0023] Preferably, the anti-inflammatory drugs use the alkaloid compounds represented by formula I or II or the pharmaceutically acceptable salts or solvates thereof as the active ingredient, and are prepared into a pharmaceutically acceptable dosage form by using a pharmaceutically acceptable carrier or excipient.

[0024] Preferably, the dosage form includes tablets, capsules, injections, aerosols, pills and ointments.

[0025] Beneficial effects:

[0026] The two alkaloid compounds with novel structures isolated from the narcissus roots in the present application can effectively inhibit the release of NO in RAW264.7 cells. As a recognized anti-inflammatory model in the art, the activity research results prove that the alkaloid compounds isolated from the narcissus roots in the present application can be used for developing anti-inflammatory drugs. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 Chemical structural formula of the compounds 1 and 2 prepared in Example 1;

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

[0029] Figure 4 , 5 HNMR spectrum of the compounds 1 and 2 1 HNMR spectrum of the compounds 1 and 2

[0030] Figure 6 、 7 HSQC spectrum of compound 1, 2; 13 C NMR spectrum;

[0031] Figure 8 、 9 HSQC spectrum of compound 1, 2;

[0032] Figure 10 、 11 HMBC spectrum of compound 1, 2;

[0033] Figure 12 、 13 NOESY spectrum of compound 1, 2;

[0034] Figure 14 、 15 H-H COSY spectrum of compound 1, 2; 1 H- 1 H COSY spectrum. DETAILED DESCRIPTION

[0035] The essential content of the present application will be specifically described below in combination with examples, but the protection scope of the present application is not limited by this.

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

[0037] I. Preparation method

[0038] The present example provides a preparation method of two alkaloid compounds in narcissus roots, and the steps are as follows:

[0039] (1) Take 10 kg of narcissus root decoction pieces, add 10 times the volume of 95% volume fraction of ethanol aqueous solution at a ratio of 1 kg:10 L, reflux extract 2 times at 78℃, each for 2 h, filter after extraction, combine the extract, and concentrate to no alcohol taste to obtain the extract concentrate; take the extract concentrate, load it into a D-101 macroporous resin column (inner diameter 20 cm x column bed height 100 cm), and elute with 60 L of water, 60 L of 30% volume fraction of methanol aqueous solution, 60 L of 60% volume fraction of methanol aqueous solution, and 60 L of pure methanol in sequence, combine the eluate of different solvents, and concentrate at 60℃ under reduced pressure to obtain the extract of different concentration elution parts;

[0040] (2) The 60% methanol aqueous solution elution part extract (29.42 g) obtained in step (1) was dissolved in methanol, and then dry-mixed with silica gel (100-200 mesh, 2 times the mass of the extract) to form a sample, which was loaded into a silica gel (100-200 mesh) column (inner diameter 8 cm x column bed height 80 cm) for column chromatography separation, and then gradient elution was performed with dichloromethane-methanol solvents with volume ratios of 100:1, 50:1, 25:1, 10:1, 5:1, 2:1, and 0:100, respectively, and each solvent was eluted for 20 L. Thin layer chromatography was used for analysis and combination (combining eluents with the same development height), and 6 flow parts A, B, C, D, E, and F were obtained.

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

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

[0043] II. Structure confirmation

[0044] Structure analysis of the compound: The structure was identified mainly by spectroscopic techniques, including mass spectrometry, nuclear magnetic resonance (H-NMR, C-NMR, 2D-NMR), and the like. 1 13 Figures 2-15

[0045] 1. Structure analysis of target compound 1

[0046] Compound 1 is a yellow powder, and the high-resolution mass spectrum ESI-TOF-MS is 326.1384 [M+H] + . The molecular formula of compound 1 is C​​​19 H 19 NO4, exact mass 326.1384, unsaturation 11.

[0047] The1H-NMR spectrum of compound 1 shows a signal peak at δ 1 H-NMR spectrum of compound 1 shows a signal peak at δ H 2.82 (3H, s) which is assigned to a N-methyl signal, δ H 3.08 (1H, dd, J = 9.8, 1.8 Hz, H-6), δ H 3.83 (1H, dd, J = 9.6, 1.8 Hz, H-6) which is assigned to two different chemically environment protons on the methylene group, δ H 3.47 (3H, s) which is assigned to a methoxy signal, δ H 4.47 (1H, d, J = 7.8 Hz, H-3) which is assigned to a methine proton signal, the aromatic region shows 7 aromatic ring protons, of which δ H 6.64 (1H, d, J = 6.0 Hz, H-5"), δ H 6.82 (1H, d, J = 1.8 Hz, H-2"), δ H 6.83 (1H, dd, J = 7.8, 1.8 Hz, H-6") which is assigned to a proton signal on the 1,3,4-trisubstituted benzene ring, δ H 6.65 (2H, d, J = 1.8 Hz, H-3', 5'), δ H 6.98 (2H, d, J = 8.4 Hz, H-2', 6') which is assigned to a proton signal on the other benzene ring which is para-disubstituted, δ H 7.20 (1H, d, J = 2.4 Hz, H-4) which is assigned to an olefinic proton signal based on the chemical shift value. In 13 The13C-NMR spectrum shows 19 carbon signals in the low field region δ C 169.0 (C-2) which is a carbonyl carbon signal, from the DEPT spectrum it can be assigned to 2 methyl, 1 methylene, 9 methine, 6 quaternary carbon signals (Table 1). In combination with DEPT, HSQC and HMBC spectra, δ C 156.5 (C-4') and δ C 126.3 (C-1') which is a quaternary carbon on the para-disubstituted benzene ring, δ C 128.2 (C-2', 6') and δ C 116.0 (C-3', 5') which is a tertiary carbon on the para-disubstituted benzene ring. δ C 147.8 (C-3"), δ C 147.7 (C-4") and δ C131.2 (C-1") is the signal of three carbons 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 signal of carbon on the 1,3,4-trisubstituted benzene ring. The combination of 1 H-NMR data, δ C 131.2 (C-4) is the signal of one carbon in the double bond. The deduced δ C 56.6 (C-6) is the signal of methylene carbon at C-6 position, the deduced δ C 55.6 is the signal of methoxy carbon on the 1,3,4-trisubstituted benzene ring, the deduced δ C 30.1 is the signal of the nitrogen methyl carbon. In 1 H- 1 In the H COSY spectrum, H-3 (δ H 4.47, 1H, d, J = 7.8 Hz) is directly correlated with H-4 (δ H 7.20, 1H, d, J = 2.4 Hz) (forming a spin-coupled system), H-3 (δ H 4.47, 1H, d, J = 7.8 Hz) is correlated with H-6 (δ H 3.08, 1H, dd, J = 9.8, 1.8 Hz, δ H 3.83, 1H, dd, J = 9.6, 1.8 Hz) and H-4 (δ H 7.20, 1H, d, J = 2.4 Hz) is correlated with H-6 (δ H 3.08, 1H, dd, J = 9.8, 1.8 Hz). In combination with the HMBC spectrum, N-CH3 is correlated with C-2, C-6, indicating that C-2, C-6, N-CH3 are connected through the nitrogen atom, H-3 is correlated with C-2, C-4, C-5 and C-6, H-4 is correlated with C-2, C-3 and C-5, H-6 is correlated with 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 the two ends are connected with the nitrogen atom. In the HMBC spectrum, H-2' / H-6' is correlated with C-3, C-4', H-3' / H-5' is correlated with C-1', C-4', and H-3 is correlated with C-1' and C-2' / C-6', indicating that C-1' on the 1,3-disubstituted benzene ring is connected with C-3. H-2" / H-6" is correlated with C-5, H-4 and H-6 are correlated with C-1", indicating that C-1" on the 1,3,4-trisubstituted benzene ring is connected with C-5. O-CH3 is correlated with C-2", C-3" / C-4", indicating that O-CH3 is connected with C-3". According to the above data, the planar structure of compound 1 can be determined asFigure 1 The compound was shown in Fig. 1 and named as 5-(4-hydroxy-3- methoxyphenyl)-3-(4-hydroxyphenyl)-N-methyl-3,6-dihydropyridin-2(lH)-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 = 8.4 Hz, H-2', 6'), δ H 6.83 (1H, dd, J = 7.8, 1.8 Hz, H-6”), δ H 6.82 (1H, d, J = 1.8 Hz, H-2”), δ H 6.65 (2H, d, J = 1.8 Hz, H-3', 5'), δ H 6.64 (1H, d, J = 6.0 Hz, H-5”), δ H 4.47 (1H, d, J = 7.8 Hz, H-3), δ H 3.83 (1H, dd, J = 9.6, 1.8 Hz, H-6), δ H 3.47 (3H, s, O-CH3), δ H 3.08 (1H, dd, J = 9.8, 1.8 Hz, H-6), δ H 2.82 (3H, s, N-CH3). 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-CH3), 40.5 (C-3), 30.1 (N-CH3).

[0049] 2. Structure analysis of target compound 2

[0050] Compound 2 was a yellow powder, and the high resolution mass spectrum ESI-TOF-MS: 296.1282 [M+H] + . The molecular formula of compound 2 was C 18 H 17 NO3, the accurate molecular weight was 296.1282, and the unsaturation degree was 11.

[0051] Compound 2 1 H-NMR, 13 The C-NMR spectrum (Table 1) was very similar to that of compound 1, with the exception of the absence of a methoxy signal for compound 2. By combining DEPT, HMQC, HMBC, 1 H- 1 The structure of the compound was determined by H-COSY spectrum as Figure 1 Compound 2 was named 3,5-di(hydroxyphenyl)-N-methyl-3,6-dihydropyridin-2(lH)-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 = 8.4 Hz, H-2', 6'), δ H 7.18 (1H, d, J = 8.4 Hz, H-6"), δ H 7.18 (1H, d, J = 8.4 Hz, H-2"), δ H 6.63 (2H, d, J = 8.4 Hz, H-3', 3"), δ H 6.61 (2H, d, J = 8.4 Hz, H-5', 5"), δ H 4.41 (1H, d, J = 7.8 Hz, H-3), δ H 3.81 (1H, dd, J = 9.6, 1.2 Hz, H-6), δ H 3.09 (1H, dd, J = 10.2, 1.8 Hz, H-6), δ H 2.81 (3H, s, N-CH3). 13 CNMR (151 MHz, 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-l"), 128.2 (C-2', 6'), 125.8 (C-l'), 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 the new compound

[0054] I. Experimental materials and instruments

[0055] Compound 1, 2 prepared in Example 1, mouse monocyte macrophage RAW264.7 (Shanghai Cell Bank of Chinese Academy of Sciences), DMEM complete medium (Beijing Solabio Technology Co., Ltd.), fetal bovine serum (FBS) (Beijing Solabio Technology Co., Ltd.), lipopolysaccharide (LPS) (Beijing Solabio Technology Co., Ltd.), DMSO (Shanghai Yisen Biological Technology Co., Ltd.), NO detection kit (Shanghai Biyun Tian Biological Technology Co., Ltd.), phosphate buffer PBS (Beijing Solabio Technology Co., Ltd.), enzyme marker (Molecular Devices: SpectraMax i3, USA), MCO-20ACI type CO2 incubator (Thermo, USA), BS125S type one hundredth balance (Mettler-Toledo, Switzerland).

[0056] II. Experimental method

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

[0058] The compound was dissolved in dimethyl sulfoxide (DMSO) to form a 100 mM stock solution, which was stored in a-20℃ refrigerator, and then diluted to the appropriate concentration with DMEM medium containing 1% fetal bovine serum (FBS) for use. In the clean bench, 5 mg of lipopolysaccharide was dissolved in 5 mL PBS and aliquoted with a sterile centrifuge tube, 1 mL per tube was sealed with a sealing film and stored in a-20℃ refrigerator.

[0059] The logarithmic growth period of RAW264.7 cells was taken at a density of 2×10 5 cells / well in a 24-well plate, and cultured for 24 hours. The blank group and the model group were replaced with 500 μL of DMEM complete medium per well, and the drug administration group was replaced with 500 μL of DMEM complete medium containing different concentrations (3.125 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM) of compounds per well. After 2 hours, 0.5 μL of LPS solution was added to each well except the blank group, and the culture was continued for 24 hours. Then 50 μL of cell supernatant was collected from each well and replicated three times, and the absorbance at 540 nm of each well was detected by enzyme marker. The cell NO release inhibition rate was calculated according to the formula [(A 模型 -A 给药 ) / (A 模型 -A 空白 )]×100%.

[0060] III. Experimental results

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

[0062] Table 1 Inhibition of NO of LPS-induced RAW264.7 cells by compound 1 at different concentrations

[0063]

[0064] Table 2 Inhibition of NO of LPS-induced RAW264.7 cells by compound 2 at different concentrations

[0065]

[0066] The determination results show that the NO inhibitory activities of the compounds 1 and 2 provided by the embodiments are continuously enhanced with the increase of the concentration, and present a certain dose dependence.

[0067] The above examples serve to specifically introduce the essential content of the present application, but those skilled in the art should know that the protection scope of the present application should not be limited to the specific examples.

Claims

1. An alkaloid compound of Formula I or II, or a pharmaceutically acceptable salt thereof.

2. A method for preparing an alkaloid compound of formula I as described in claim 1, characterized in that, comprising the steps of: (1) taking the Zizanin root medicinal material, heating extraction with 95% ethanol aqueous solution, filtering the extract, concentrating to remove alcohol taste, to obtain extract concentrate; taking the extract concentrate, loading onto a D-101 macroporous resin column, eluting with water, 30% methanol aqueous solution, 60% methanol aqueous solution, and pure methanol in sequence, and combining the eluate of different solvents, to obtain extract of different elution fractions after vacuum concentration; (2) taking the extract of the 60% methanol aqueous solution elution fraction obtained in step (1), loading onto a silica gel column, performing column chromatography separation, then gradient eluting with dichloromethane-methanol solvents in 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 by thin layer chromatography, to obtain 6 flow fractions A, B, C, D, E, and F; (3) The stream fraction C obtained in step (2) was dissolved with methanol, and separated by preparative liquid chromatography, eluted with acetonitrile-water (18:82 by volume) isocratic, and the eluent corresponding to the chromatographic peak with a retention time of 57 min was collected and concentrated to dryness to obtain the product; the chromatographic conditions of the preparative liquid chromatography were as follows: YMC-Triart C 18 Chromatographic column, column size 250 mm*10 mm, particle size 5 μm, eluent flow rate 4 mL / min.

3. The method of claim 2, wherein: The solvent for heating extraction in step (1) is 95% ethanol aqueous solution.

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

5. A method for preparing an alkaloid compound of formula II as described in claim 1, characterized in that, comprising the steps of: (1) taking the Zizanin root medicinal material, heating extraction with 95% ethanol aqueous solution, filtering the extract, concentrating to remove alcohol taste, to obtain extract concentrate; taking the extract concentrate, loading onto a D-101 macroporous resin column, eluting with water, 30% methanol aqueous solution, 60% methanol aqueous solution, and pure methanol in sequence, and combining the eluate of different solvents, to obtain extract of different elution fractions after vacuum concentration; (2) taking the extract of the 60% methanol aqueous solution elution fraction obtained in step (1), loading onto a silica gel column, performing column chromatography separation, then gradient eluting with dichloromethane-methanol solvents in 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 by thin layer chromatography, to obtain 6 flow fractions A, B, C, D, E, and F; (3) taking the flow fraction C obtained in step (2), dissolving with methanol, and separating by preparative liquid chromatography, eluting with acetonitrile-water in a volume ratio of 18:82, collecting the eluate at 32-33 min, and concentrating and drying to obtain sub-flow fraction C-1; The 40-41 min eluate was collected, concentrated and dried to obtain sub-fraction C-2; the chromatographic conditions of preparative liquid chromatography were as follows: YMC-Triart C 18 Chromatographic column, column size 250 mm*10 mm, particle size 5 μm, eluent flow rate 4 mL / min; (4) The sub-fraction C-2 obtained in step (3) was dissolved in methanol, and then separated by preparative liquid chromatography, eluted with acetonitrile-water (10:90 by volume) as the mobile phase, and the eluate corresponding to the chromatographic peak with a retention time of 19 min was collected and concentrated to dryness to obtain the product; the chromatographic conditions of the preparative liquid chromatography were as follows: YMC-Triart C 18 Chromatographic column, column size 250 mm*10 mm, particle size 5 μm, eluent 4 mL / min.

6. The method of claim 5, wherein: The solvent for heating extraction in step (1) is 95% ethanol aqueous solution.

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

8. Use of the alkaloid compound of Formula I or II or a pharmaceutically acceptable salt thereof in claim 1 for preparing an anti-inflammatory drug.

9. Use according to claim 8, characterized in that: The anti-inflammatory drug takes the alkaloid compound of Formula I or II or a pharmaceutically acceptable salt thereof as an active ingredient, and is prepared into a pharmaceutically acceptable dosage form through a pharmaceutically acceptable carrier or excipient.

10. Use according to claim 9, characterized in that: The dosage form includes tablets, capsules, injections, aerosols, pills, and ointments.

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