New lignan compound extracted and separated from cyclocarya paliurus leaf, and preparation method and application thereof

Abstract

The present application belongs to the technical field of natural medicinal chemistry, and particularly relates to a new lignan compound extracted and separated from Cyclobalanopsis glaucoides leaves. Meanwhile, the present application also relates to a method for extracting and separating the new lignan compound from Cyclobalanopsis glaucoides leaves, which comprises the following steps: taking dried Cyclobalanopsis glaucoides leaves, crushing the leaves, and then impregnating and extracting the crushed leaves with ethanol; concentrating the extract, and then extracting the concentrated extract with petroleum ether, ethyl acetate and water-saturated n-butanol in sequence; subjecting the n-butanol extract to D101 macroporous resin, silica gel chromatographic column, Sephadex LH-20 and ODS column chromatography in sequence, and then purifying the new compound by semi-preparative HPLC. In addition, the new compound has obvious inhibitory effect on the generation of NO at a concentration of 10 muM, and the inhibition rate is 52.66%, so that the new compound has good anti-inflammatory activity.

Description

Technical Field

[0001] This invention belongs to the field of natural medicinal chemistry technology, specifically relating to a novel lignan compound extracted and isolated from the leaves of Eucommia ulmoides and its application. Background Technology

[0002] *Cyclocarya paliurus* (Batal.) Iljinsk., also known as the money tree, sweet-leaf tree, and sweet tea tree, is a deciduous tree belonging to the genus *Cyclocarya* in the family Juglandaceae. It is a monotypic genus endemic to my country and is known as the "giant panda of the plant kingdom," the "third tree of medicine," and a natural source of insulin. It is widely distributed in Central and Eastern my country, typically growing in valleys, riverbanks, or humid forests at altitudes of 500-2500 m. *Cyclocarya paliurus* is neutral in nature, pungent and slightly bitter in taste. Traditional medical records indicate that it has various effects, including clearing heat and reducing swelling, anti-inflammatory and analgesic properties, lowering blood pressure and strengthening the heart, and external treatment of stubborn tinea. Modern pharmacological research also shows that *Cyclocarya paliurus* not only has anti-inflammatory and hypoglycemic effects but also plays a significant role in lowering lipids, lowering blood pressure, and antioxidation. It can effectively prevent and treat various chronic diseases and has considerable medicinal and health benefits.

[0003] Current research on the chemical components of *Cyclocarya paliurus* mainly focuses on polysaccharides, flavonoids, and triterpenoids, with limited research on other components. The chemical composition of *Cyclocarya paliurus* is complex, containing numerous components with similar molecular weights and polarities, making the isolation of new compounds challenging. Pharmacological activity studies of *Cyclocarya paliurus* also primarily focus on extracts, with limited research on the pharmacological activity of individual chemical components. Related product development mainly involves processing the raw material or its extracts into tea bags, health tonics, and compressed candies, resulting in relatively crude processes and low economic added value. NO receptors, as important drug targets widely present in the human body, are closely related to vascular endothelial function, pancreatic function, neural regulation, and immune responses. This invention isolates a new compound from *Cyclocarya paliurus* that exhibits good pharmacological activity in inhibiting NO production, providing a material basis and new pathway for the development of new drugs related to *Cyclocarya paliurus*. Summary of the Invention

[0004] One of the objectives of this invention is to provide a novel lignan compound extracted and isolated from the leaves of *Cyclocarya paliurus*, the structure of which is shown in formula (I).

[0005] (I).

[0006] The chemical formula of the compound obtained in formula (Ⅰ) is C 27 H 36 O 12 Its name is (-)-cantharis resinol-4-O-β-D-xylanoside.

[0007] A second objective of this invention is to provide a method for preparing the novel lignan compound described in one objective of this invention, the method comprising the following steps:

[0008] (1) Extract the leaves of Cyclocarya paliurus by soaking in ethanol, concentrate the extract to obtain an ethanol extract in paste form, and suspend the ethanol extract in water to obtain a suspension.

[0009] (2) Petroleum ether, ethyl acetate and water-saturated n-butanol of the same volume as the suspension were added to the suspension in sequence for extraction. The water-saturated n-butanol extract was retained and the others were discarded. The water-saturated n-butanol extract was concentrated to a paste to obtain the water-saturated n-butanol extract fraction.

[0010] (3) The water-saturated n-butanol extraction fraction was added to a macroporous adsorption resin chromatography column and eluted sequentially with water and 30% ethanol. The eluent was discarded, and then eluted with 60% ethanol. The eluent was collected and evaporated to dryness to obtain sample A.

[0011] (4) Sample A was added to a 10% deactivated silica gel chromatography column and eluted sequentially with chloroform-methanol at a volume ratio of 10:1 and chloroform-methanol at a volume ratio of 7:1. The eluent was discarded, and then eluted with chloroform-methanol at a volume ratio of 10:3. The eluent was collected and evaporated to dryness to obtain sample B.

[0012] (5) Sample B was added to a dextran gel chromatography column, eluted with methanol, and monitored by thin-layer chromatography to obtain two subfractions, B1 and B2.

[0013] (6) Sample B2 was subjected to Flash fast ODS column chromatography and eluted with a methanol-water gradient at a volume ratio of 10:100-100:0 to obtain two subfractions, sample B2a and B2b.

[0014] (7) The B2b was eluted by semi-preparative HPLC isocratic elution to obtain the compound of formula (Ⅰ).

[0015] In a preferred embodiment of the present invention, in step (1), the mass ratio of the Eucommia ulmoides leaves to the ethanol is 1:6~12.

[0016] In a preferred embodiment of the present invention, in step (1), the concentration of ethanol is 70% to 80%.

[0017] In a preferred embodiment of the present invention, in step (1), the immersion extraction is performed three times, each time for three days.

[0018] In a preferred embodiment of the present invention, in step (2),

[0019] The petroleum ether extraction was performed three times; and / or

[0020] The ethyl acetate was extracted three times; and / or

[0021] The water-saturated n-butanol was extracted three times.

[0022] In a preferred embodiment of the present invention, in step (7), the semi-preparative HPLC elution conditions are acetonitrile-water with a volume ratio of 18:82, a flow rate of 10.0 mL / min, and a detection wavelength of 203 nm.

[0023] In a preferred embodiment of the present invention

[0024] In step (3), the macroporous adsorption resin in the macroporous adsorption resin chromatography column is type D101; and / or

[0025] In step (5), the dextran gel chromatography column is Sephadex LH-20.

[0026] A third objective of this invention is to provide the application of the novel lignan compound described in one objective of this invention in the preparation of anti-inflammatory drugs.

[0027] Compared with the prior art, the present invention has the following advantages:

[0028] This invention extracts and isolates a novel compound, (-)-cantharides resinol-4-O-β-D-xylanoside, from the leaves of *Cyclocarya paliurus*. Furthermore, this invention evaluates the anti-inflammatory activity of this novel compound at the cellular level. The compound significantly inhibits NO production at a concentration of 10 μM, with an inhibition rate of 52.66%, indicating that the compound possesses good anti-inflammatory activity. Attached Figure Description

[0029] Figure 1 These are the HPLC spectra of the derivative of the new compound obtained in Example 1 of this invention and the derivative of the reference standard.

[0030] Figure 2 This is the UV spectrum of the new compound obtained in Example 1 of this invention.

[0031] Figure 3 This is the IR spectrum of the new compound obtained in Example 1 of the present invention.

[0032] Figure 4 This is the HR-TOF-MS spectrum of the new compound obtained in Example 1 of this invention.

[0033] Figure 5 This is the new compound obtained in Example 1 of the present invention. 1 H-NMR spectrum.

[0034] Figure 6 This is the new compound obtained in Example 1 of the present invention. 13C-NMR spectrum.

[0035] Figure 7 This is the HSQC spectrum of the new compound obtained in Example 1 of this invention.

[0036] Figure 8 This is the new compound obtained in Example 1 of the present invention. 1 H- 1 H-COSY spectrum.

[0037] Figure 9 This is the HMBC spectrum of the new compound obtained in Example 1 of this invention.

[0038] Figure 10 This is the NOESY spectrum of the new compound obtained in Example 1 of this invention.

[0039] Figure 11 This is the CD spectrum of the new compound obtained in Example 1 of this invention. Detailed Implementation

[0040] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments and accompanying drawings, so that those skilled in the art can better understand the present invention, but this does not limit the present invention.

[0041] In this invention, unless otherwise specified, all percentages are volume percentages.

[0042] In this invention, RAW 264.7 refers to mouse mononuclear macrophages; LPS stands for Lipopolysaccharide.

[0043] Example 1 This example involves the extraction, separation, and structural confirmation of compounds.

[0044] Step 1: Take 49.59 kg of dried Eucommia ulmoides leaves, pulverize them, and add 80% ethanol at a material-to-liquid ratio of 1:6. Extract three times by maceration, each time for three days. Combine the extracts and concentrate under reduced pressure until there is no alcohol odor to obtain a paste-like ethanol extract (9.383 kg).

[0045] Step 2: Add 20L of purified water to the paste-like ethanol extract for dispersion, and extract three times each with equal volumes of petroleum ether, ethyl acetate and water-saturated n-butanol. Retain the water-saturated n-butanol extract and discard the others. Concentrate the water-saturated n-butanol extract to a paste-like state to obtain the water-saturated n-butanol extract fraction (1.333 kg).

[0046] Step 3: The water-saturated n-butanol fraction was subjected to D101 macroporous adsorption resin column chromatography, eluted sequentially with water and 30% ethanol until the eluent was colorless. The eluent was discarded, and then eluted with 60% ethanol for 2 column volumes. The eluent was collected and evaporated to dryness to obtain sample A (183.56g).

[0047] Step 4: Add sample A to a 10% deactivated silica gel chromatography column and elute sequentially with chloroform-methanol at a volume ratio of 10:1 and 7:1 until the eluent is colorless. Discard the eluent and then elute with chloroform-methanol at a volume ratio of 10:3. Collect the eluent and evaporate it to dryness to obtain sample B (5.8 g).

[0048] Step 5: Sample B is added to a dextran gel chromatography column and eluted isocratically with methanol. Thin-layer chromatography is used to monitor the eluent, yielding two subfractions: sample B1 (eluted after 0.7-1.0 column volumes) and B2 (eluted after 1.2-1.4 column volumes). The dextran gel chromatography column is a Sephadex LH-20.

[0049] Step 6: Sample B2 was subjected to Flash rapid ODS column chromatography with a methanol-water gradient elution of 10:100-100:0 (v / v). Specifically, starting with 10:100 methanol-water, the methanol concentration was increased by 5% per minute until 100% methanol was reached, followed by continuous elution with 100% methanol for 20 minutes. The entire elution process took 38 minutes. The UV absorbance of the eluent and sample was measured at 254 nm, and two subfractions, B2a and B2b, were collected sequentially. The column type was Flash pureID C18; the column volume was 114 ml; and the flow rate was set to 60 ml / min.

[0050] Step 7: After isocratic elution of B2b by semi-preparative HPLC, a new compound was obtained, with a retention time range of 25.2-26.5 min. The semi-preparative HPLC elution conditions were as follows: eluent was 18% acetonitrile-water solution, flow rate was 10.0 mL / min, detection wavelength was 203 nm; the chromatographic column was a COSMOSIL 5C18-AR-II with dimensions of 250 × 20 mm and 5 μm.

[0051] The new compound obtained in Example 1 was a white amorphous powder (5.7 mg). The structure of the new compound obtained in Example 1 was identified as follows: First, the specific rotation of the compound obtained in Example 1 was measured, and the result was [α]20 D -26.0° (c 0.1, MeOH). Second, the compound obtained in Example 1 was subjected to a vanillin-concentrated sulfuric acid colorimetric test, and the result showed a pink color. Third, after acid hydrolysis and derivatization, the compound obtained in Example 1 was analyzed by HPLC and found to contain D-xylose (see [link to example]). Figure 1 It should be noted that... Figure 1 The lower curve corresponds to the D-xyloside acid hydrolysis derivative, while the upper curve corresponds to the acid hydrolysis derivative of the compound obtained in Example 1. Finally, UV, IR, HR-TOF-MS, and other methods were used to analyze the results. 1 HNMR, 13 CNMR and two-dimensional nuclear magnetic resonance spectroscopy (HSQC, HMBC, ... 1 H- 1 The structures of the new compounds obtained in Example 1 were identified by H-COSY, NOSEY, and CD, and the results are as follows: Figures 2-11 As shown.

[0052] The method for testing optical rotation is as follows: Take an appropriate amount of the sample to be tested, prepare a test solution with methanol to a concentration of 0.1 g / L, inject the test solution into the test tube, place it in a polarimeter to detect the reading, and calculate the specific rotation. The method for testing the vanillin-concentrated sulfuric acid reaction is as follows: Weigh 1.0 g of vanillin and dissolve it in concentrated sulfuric acid to prepare a solution with a concentration of 10 g·L. -1 The vanillin-concentrated sulfuric acid colorimetric reagent was used. The sample solution for optical rotation determination was spotted onto a silica gel plate, and the vanillin-concentrated sulfuric acid colorimetric reagent was sprayed onto the spotted area. The acid hydrolysis derivatization method was as follows: 1.0 mg of the new compound obtained in Example 1 was accurately weighed and added to a hydrochloric acid solution (0.6 mL, 0.5 mol / L). The mixture was reacted at 90 °C for 2 h to allow complete hydrolysis. After hydrolysis, anion exchange resin IRA 400 was added to adjust the pH to neutral. The resin was removed by filtration, and the filtrate was concentrated under reduced pressure and dried under vacuum. 0.2 mL of a pyridine solution containing 1.0 mg / L cysteine ​​methyl ester hydrochloride was added to the filtrate residue, and the mixture was reacted at 60 °C for 1 h. Then, 0.2 mL of a pyridine solution containing 1.0 mg of o-toluene isothiocyanate was added to the reactants, and the mixture was reacted at 60 °C for 1 h to obtain the derivative of the sugar fraction in the sample. The sugar standard was also derivatized under the same conditions as a reference derivative. The acid-hydrolyzed derivatives (as test solutions) and reference derivatives (as reference solutions) of the new compound obtained in Example 1 were both injected into HPLC for detection. The results are as follows: Figure 1 As shown. According to Figure 1 It can be seen that the peak elution times of the test solution and the reference solution are consistent, thus confirming that D-xylose is present in the new lignan compound obtained in Example 1. Figures 2-11 The testing method used in the Chinese Spectrum is a standard method in this field and will not be described in detail here.

[0053] Figure 2 The UV spectrum of the compound in Example 1 is shown. Figure 2 It can be seen that its maximum absorption wavelength is 214 nm. Figure 3 The IR spectra of the compounds in Example 1 are shown. Figure 3 It can be seen that hydroxyl groups are present in the IR spectrum (3376.4 cm⁻¹). -1 ), aromatic ring C=C bond (1600.7 cm) -1 ) and CO bonds (1076.9, 1032.9 cm) -1 Absorption peak. Figure 4 The HR-TOF-MS spectra of the compounds in Example 1 are shown. Figure 4 The quasi-molecular ion peak m / z is 575.20894 [M+Na]. + (C 27 H 36 O 12 Na, calcd. 575.2099), thus the molecular formula of the new compound obtained in Example 1 can be determined to be C. 27 H 36 O 12 Its molecular weight is 552.

[0054] Figure 5 Table 1 shows the compounds in Example 1. 1 H NMR spectrum information, from Figure 5 As shown in Table 1, it displays a set of 1,2,3,4,5-pentasubstituted aromatic ring hydrogen signals δ 6.71 (1H, s), a set of 1,3,4,5-tetrasubstituted aromatic ring hydrogen signals δ 6.44 (2H, s); a sugar terminal hydrogen signal δ 5.01 (1H, d, J = 6.6 Hz); four singlet methoxy hydrogen signals δ 3.90, 3.80×2, and 3.41; and other hydroxyl hydrogen signals δ 3.96 ~ 3.19. Figure 6 Table 1 shows the compounds in Example 1. 13 C NMR spectrum information, from Figure 6 As shown in Table 1, this compound has 27 carbon signals, of which δ 153.4, 153.3, 149.1×2, 139.2, 138.0, 136.2, 134.6, 126.8, 108.8, and 106.9×2 represent twelve aromatic ring carbon signals, δ 104.7 represents the sugar terminal carbon signal, δ 76.7 ~ 66.7 represent oxygen-bound carbon signals on the sugar ring, δ 66.5 and 63.9 represent two oxygen-bound methylene carbon signals, and δ 61.1, 56.8×2, and 56.7 represent four oxygen-bound methyl carbon signals. (Combined with...) Figure 5 and Figure 6 middle 1 H NMR, 13 According to the C18 NMR data, the compound is a 2,7'-cyclolignan compound. Combined with the aforementioned acid hydrolysis results, it is known that the compound in Example 1 contains a β-D-xylose.

[0055] Further determination of the carbon and hydrogen correlation signals of this new compound was made based on 2D NMR, and in Figure 9 The HMBC spectrum was used to determine the connection positions and order of sugars in the structure. This was combined with the δ values ​​in the HMBC spectrum. H-1'' 5.01 and δ C-4 The correlation signal at 138.0 further confirmed that xylose is linked at the C-4 position of the aglycone. In the NOESY spectrum (e.g.) Figure 10 ), δ H-7a 2.80, δ H-7' 4.36 and δ H-8 The presence of a relevant signal at 1.70 indicates that H-8 and H-7' are β configurations; δ H-7b 2.68 and δ H-8' A signal at 2.03 indicates that H-8' is in the α configuration. Furthermore, the absolute configuration at the C-7' position can be determined by circular dichroism (CD) (e.g., Figure 11 The determination is made by [the relevant authority / organization]. Figure 11 It can be seen that the compound in Example 1 exhibits a positive Cotton effect at 241 nm, proving that the C-7' position is in the R configuration.

[0056] Table 1. NMR data of the new compound (CD3OD, J in Hz)

[0057] Position delta H a) ]]> C b) ]]> ​ <![CDATA[ 1 H- 1 H COSY]]> HMBC (H→C) Agly 1 — 136.2 — — 2 — 126.8 — — 3 — 153.3 — — 4 — 138.0 — — 5 — 153.4 — — 6 6.71 (s) 108.8 — C-1, 2, 4, 5, 7 7 2.79 (dd, 15.3, 4.6), 2.68 (dd, 15.3, 11.3) 33.8 H-8 C-1, 2, 6, 8, 9, 8' 8 1.70 (m) 40.6 H-7, 9, 8' — 9 <![CDATA[3.67 (dd, 11.0, 5.0), 3.57 c) ]]> 66.5 H-8 C-7, 8, 8' 3-OMe 3.41 (s) 61.1 — C-3 5-OMe 3.90 (s) 56.7 — C-5 1' — 139.2 — — 2' 6.44 (s) 106.9 — C-1', 3', 4', 6', 7' 3' — 149.1 — — 4' — 134.6 — — 5' — 149.1 — — 6' 6.44 (s) 106.9 — C-1', 2', 4', 5', 7' 7' 4.36 (d, 5.7) 42.4 H-8' C-1, 2, 3, 8, 1', 8', 9' 8' 2.03 (m) 48.7 H-8, 7', 9' C-1, 2, 8, 9, 7', 9' 9' <![CDATA[3.57 c) ]]> 63.9 H-8' C-8, 7', 8' 3'-OMe 3.80 (s) 56.8 — C-3' 5'-OMe 3.80 (s) 56.8 — C-5' Xyl 1'' 5.01 (d, 6.6) 104.7 H-2'' C-4, 5'' 2'' <![CDATA[3.55 c) ]]> 74.8 H-1'', 3'' C-1'', 3'' 3'' 3.46 (t, 8.3) 76.7 H-2'', 4'' C-2'', 4'' 4'' 3.61 (br dd, 8.3,4.9) 71.1 H-3'', 5'' C-3'' 5'' 3.96 (dd, 11.7, 4.9), 3.19 (dd, 11.7, 8.8) 66.7 H-4'' C-1'', 3'', 4''

[0058] a) Measured at 500 MHz; b) Measured at 125 MHz; c) Overlapped signals;Xyl: xylopyranosyl.

[0059] Based on the above analysis, the results were obtained through vanillin-concentrated sulfuric acid colorimetric test, UV, IR, HR-TOF-MS, 1 H NMR, 13 CNMR and two-dimensional nuclear magnetic resonance spectroscopy (HSQC, HMBC, ... 1 H- 1 H-COSY, NOSEY, CD) confirmed that the structure of the compound in Example 1 is (-)-lyoniresinol-4-O-β-D-xylopyranoside [(-)-lyoniresinol-4-O-β-D-xylopyranoside], and its chemical structural formula is as follows:

[0060]

[0061] (I).

[0062] Example 2 This example relates to the pharmacological activity testing of the new compound obtained in Example 1.

[0063] Take a RAW264.7 cell cryopreservation tube and place it in a 37°C water bath until the cryopreservation solution thaws. In a biosafety cabinet, transfer the cryopreservation solution to a centrifuge tube, centrifuge, and discard the supernatant. Add 2 mL of DMEM complete culture medium, disperse the cells, and evenly spread them in a culture dish. Incubate in a cell culture incubator. When the cells reach 80%-90% confluence, passage them for optimal cell condition and then plate them.

[0064] Take healthy RAW264.7 cells and use 1×10⁻⁶ cells. 4 Cells were seeded per well in 96-well plates and tested when the cells reached 60% confluence. The experiment consisted of four groups: a blank control group, a model group, a positive control group, and a drug-treated group. The blank control group was cultured in DMEM complete medium containing 10% fetal bovine serum. The drug-treated group received 10 μM of the novel lignan compound obtained in Example 1, followed by the addition of LPS to a final concentration of 1 μg / mL 2 hours later. The positive control group received 10 μM indomethacin, followed by the addition of LPS to a final concentration of 1 μg / mL 2 hours later. The model group also received LPS to a final concentration of 1 μg / mL, with the addition time the same as the drug-treated and positive control groups. All four groups of cells were incubated for 24 hours, with three replicates per group. After 24 hours, 50 μL of supernatant from each group was transferred to a 96-well cell culture plate, and the appropriate reagent was added according to the Griess reagent method, gently shaking several times to ensure thorough mixing. The absorbance A of the four groups was measured at a wavelength of 540 nm using an enzyme-linked immunosorbent assay (ELISA) reader, and the NO content was calculated based on the standard curve.

[0065] The results showed that after 24 hours of LPS stimulation, the NO content in the model group was 42.68 μmol / L, significantly higher than that in the control group (11.85 μmol / L), indicating successful establishment of the cell model. After drug treatment, the NO content in the model group was 26.43 μmol / L, and in the positive control group it was 26.09 μmol / L, both showing significant inhibitory effects on NO production. The inhibition rate in the drug-treated group was 52.66%, and in the positive control group it was 53.81%. Therefore, the novel lignan compound of this invention possesses good anti-inflammatory activity.

Claims

1. A new compound of lignan extracted and separated from Cyclobalanopsis glauca, characterized in that: The structure of the lignan new compound is shown as formula (I) ​ (Ⅰ)。 2. A method for preparing the novel lignan compound according to claim 1, characterized by: The method comprises the following steps: (1) extracting the leaves of Adenocarpus foliolatus with ethanol to obtain an ethanol extract in the form of extract, and suspending the ethanol extract in water to obtain a suspension; (2) sequentially adding petroleum ether, ethyl acetate and water-saturated n-butanol to the suspension to extract, retaining the water-saturated n-butanol extract and discarding the others; and concentrating the water-saturated n-butanol extract to the form of extract to obtain a water-saturated n-butanol extract part; (3) loading the water-saturated n-butanol extract part into a macroporous adsorption resin chromatographic column, sequentially eluting with water and 30% ethanol, discarding the eluate, then eluting with 60% ethanol, collecting the eluate and evaporating to dryness to obtain sample A; (4) loading the sample A into a 10% inactivated silica gel chromatographic column, sequentially eluting with chloroform-methanol (10:1 by volume ratio), chloroform-methanol (7:1 by volume ratio) and chloroform-methanol (10:3 by volume ratio), discarding the eluate, then eluting with chloroform-methanol (10:3 by volume ratio), collecting the eluate and evaporating to dryness to obtain sample B; (5) loading the sample B into a dextran gel chromatographic column, isocratically eluting with methanol, and monitoring by thin layer chromatography to obtain two subfractions, sample B1 and B2; (6) loading the sample B2 into a Flash rapid ODS column, gradient eluting with methanol-water (10:100-100:0 by volume ratio) to obtain two subfractions, sample B2a and B2b; (7) isocratically eluting the B2b by semi-preparative HPLC to obtain the compound of formula (I).

3. The method of claim 2, wherein, In step (1), the mass ratio of the leaves of Adenocarpus foliolatus to the ethanol is 1:6-12.

4. The preparation method according to claim 2, characterized in that, In step (1), the concentration of the ethanol is 70%-80%.

5. The preparation method according to claim 2, characterized in that, In step (1), the extraction is performed for three times, each for three days.

6. The preparation method according to claim 2, characterized in that, In step (2), the extraction with petroleum ether is performed for 3 times; and / or the extraction with ethyl acetate is performed for 3 times; and / or the extraction with water-saturated n-butanol is performed for 3 times.

7. The method of claim 2, wherein: In step (7), the semi-preparative HPLC elution condition is that the eluent is 18% acetonitrile-water solution, the flow rate is 10.0 mL / min, and the detection wavelength is 203 nm.

8. The preparation method according to claim 2, characterized in that: in step (3), the macroporous adsorption resin in the macroporous adsorption resin chromatographic column is D101 type; and / or in step (5), the dextran gel chromatographic column is Sephadex LH-20.

9. Use of the lignan new compound according to claim 1 in the preparation of an anti-inflammatory drug.

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