Erimophyllone-type sesquiterpenoids, methods of making, and uses thereof

Abstract

The application discloses an eriomol-type sesquiterpene compound, which is extracted from roots of Lindera aggregata, and is separated and purified by a silica gel column chromatography and the like. The two obtained compounds have good anti-proliferation activity on breast cancer cells, and can be used for preparing anti-tumor compounds.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more specifically, to two novel eromethoate-type sesquiterpenoid compounds from Lindera root, their preparation methods, and their application in the preparation of antitumor drugs. Background Technology

[0002] Breast cancer is a common malignant tumor in women. Despite extensive research on breast cancer by clinicians and basic scientists, the treatment outcomes for breast cancer patients remain unsatisfactory. Personalized treatment for breast cancer patients is one of the important principles of cancer treatment. Surgical resection, radiotherapy, chemotherapy, and targeted drug therapy have been successfully applied to the treatment of breast cancer patients. However, surgical resection is more effective for early-stage breast cancer, but 75-80% of patients with lymph node metastasis will experience recurrence or metastatic breast cancer within 10 years and eventually die from breast cancer. At the same time, surgical resection is not effective for patients with metastatic breast cancer. Radiotherapy and chemotherapy have significant side effects, which seriously affect the quality of life of breast cancer patients. Therefore, traditional Chinese medicine is increasingly gaining widespread attention in the treatment of breast cancer patients. For example, ginsenoside R3 can inhibit the proliferation and metastasis of estrogen-positive human breast cancer cells MCPF-7 and induce their apoptosis, inhibiting the invasive ability of MCF-7 cells (Chen Yun, Zhang Bing, Yu Yong. Effects of ginsenoside Rg3 on the proliferation and invasion of breast cancer MCF-7 cells [J]. Chinese Journal of Modern Applied Pharmacy, 2013, 30(7): 722-725).

[0003] Lindera aggregata (Sims) Kosterm, also known as Tiantai Lindera, Copper Coin Tree, Fragrant Leaf, and Fragrant Wood, is a plant belonging to the genus Lindera in the Lauraceae family. It is abundant in my country, mainly distributed in Zhejiang, Anhui, Hunan, Guangdong, and Guangxi provinces, and has significant development value. In 2018, due to its strong regional characteristics, outstanding therapeutic effects, and large-scale production, Lindera was designated as one of the new "Eight Treasures of Zhejiang". The medicinal part of Lindera is its spindle-shaped tuberous root, which is darker in the center, fragrant, pungent, and warm in nature, and enters the lung, spleen, kidney, and bladder meridians. The earliest record of Lindera's medicinal use is found in the Song Dynasty's *Kaibao Materia Medica*, where it was used internally to treat nausea, abdominal pain, indigestion, enuresis, and frequent urination. In addition, Lindera also has the effect of soothing the liver, regulating qi, and relieving depression. Formulas such as Tianhe Lindera Powder, Simo Decoction, and Nuangan Decoction use Lindera, which enters the Jueyin Liver Meridian, to regulate qi, soothe the liver, warm the liver, and dispel cold, serving as the principal ingredient. Lindera root, used in the Shugan Sanyu Decoction (Liver-Soothing and Blood-Stasis-Dispersing Decoction) primarily for treating liver cancer, blood stasis, and stabbing pain in the hypochondriac region, also plays a role in soothing and nourishing the liver. Modern natural product chemistry research shows that Lindera root possesses anti-tumor, anti-inflammatory, antibacterial, anticoagulant, and hepatoprotective activities, and its secondary metabolites have great development potential. The erymophenol-type sesquiterpenoid compounds (1, 2) derived from Lindera root involved in this invention have good anti-tumor effects and can be used to prepare anti-tumor drugs. Summary of the Invention

[0004] The purpose of this invention is to provide erymophenol-type sesquiterpenoid compounds (1, 2) from Lindera root, their preparation methods, and their application in antitumor drugs.

[0005] To achieve the above objectives, the present invention adopts the following technical solution.

[0006] In a first aspect, the present invention provides an irimophenol-type sesquiterpene compound, wherein the irimophenol-type sesquiterpene compound is one of the following:

[0007]

[0008] Preferably, the erymophenol-type sesquiterpene compound is the compound shown in Formula 1.

[0009] Secondly, this invention also provides a method for preparing the aforementioned erymophenol-type sesquiterpenoid compounds. The sesquiterpenoid compounds 1 and 2 of this invention are extracted from the roots of Lindera aggregata (Sims) Kosterm. Lindera aggregata, the dried tuberous root of the Lauraceae plant Lindera aggregata, is widely distributed in southern China and has the effects of regulating qi and relieving pain, warming the kidneys and dispelling cold. The specific method is as follows:

[0010] (1) Extract the dried Lindera aggregata (Sims) Kosterm root fragments (preferably Lindera root powder obtained by crushing and sieving (80 mesh)) with a 95% ethanol aqueous solution, filter, and concentrate the filtrate under reduced pressure to obtain a crude extract.

[0011] (2) Disperse the crude extract obtained in step (1) evenly in water, extract with ethyl acetate, combine the ethyl acetate layers, concentrate under reduced pressure to obtain ethyl acetate extract;

[0012] (3) The ethyl acetate extract obtained in step (2) was subjected to silica gel column chromatography. The silica gel column chromatography was performed by gradient elution with a mixed solvent of petroleum ether and ethyl acetate in volume ratios of 15:1, 10:1, 8:1, 6:1, 4:1, 2:1 and 1:1, respectively. Each gradient elution was performed for 2 to 3 column volumes. The elution fractions in volume ratios of 4:1 and 2:1 were collected and concentrated under reduced pressure to obtain component A and component B.

[0013] (4) Take component B from step (3) and elute it with a mixed solvent of methanol and water in a volume ratio of 50:50 using an MCI column. Use a mixed solvent of chloroform and methanol in a volume ratio of 30:1 as the developing solvent for TLC detection. Combine the fractions with an Rf value of 0.3 and concentrate under reduced pressure to obtain concentrate B1.

[0014] Concentrate B1 was subjected to silica gel column chromatography, with gradient elution using a mixed solvent of petroleum ether and ethyl acetate at volume ratios of 5:1, 4:1, 3:1, 2:1, and 1:1, two column volumes for each gradient. TLC was performed using a mixed solvent of chloroform and methanol at a volume ratio of 30:1 as the developing solvent. Fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate B2.

[0015] The concentrate B2 was dissolved in methanol and subjected to ODS C-18 column chromatography. The ODS C-18 column chromatography was performed by gradient elution with mixed solvents of methanol and water in volume ratios of 30:70, 40:60, 50:50, 60:60, 70:30, 80:20, 90:10, and 100:0, with each gradient eluting two column volumes. The fractions were analyzed by TLC with a mixed solvent of chloroform and methanol in a volume ratio of 30:1. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain the erymophenol-type sesquiterpene compound shown in Formula 1.

[0016] (5) Take component A from step (3) and use an MCI column to perform gradient elution with a mixed solvent of methanol and water in volume ratios of 30:70, 40:60, 50:50, 60:60, 70:30, 80:20, 90:10, and 100:0. Each gradient elution is two column volumes. Use a mixed solvent of chloroform and methanol in a volume ratio of 50:1 as the developing solvent for TLC detection. Combine the fractions with an Rf value of 0.3 and concentrate under reduced pressure to obtain concentrate A1.

[0017] The concentrate A1 was subjected to silica gel column chromatography with a mixed solvent of petroleum ether and ethyl acetate in a volume ratio of 10:1 as the eluent and TLC detection with a mixed solvent of chloroform and methanol in a volume ratio of 50:1 as the developing solvent. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate A2.

[0018] The concentrate A2 was dissolved in methanol and subjected to ODS C-18 column chromatography with a methanol-water mixture of 40:60 (v / v) as the eluent and a chloroform-methanol mixture of 50:1 (v / v) as the developing solvent. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate A3.

[0019] The concentrate A3 was subjected to silica gel column chromatography using a mixed solvent of chloroform and methanol at a volume ratio of 100:1 as the eluent, and TLC was performed using a mixed solvent of chloroform and methanol at a volume ratio of 50:1 as the developing solvent. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate A4.

[0020] The concentrate A4 was eluted by semi-preparative high-performance liquid chromatography using a methanol-water mixture (79:21 v / v) as the eluent, and t was collected. R The eluent fraction of 22.3 min was concentrated under reduced pressure to obtain the erymophenol-type sesquiterpene compound shown in Formula 2.

[0021] Furthermore, the volume of the ethanol-water solution in step (1) is 5 L / kg based on the mass of the Lindera root.

[0022] The extraction operation described in step (1) of this invention is as follows: at room temperature (20-30℃), immerse the fragments of Lindera root in the ethanol aqueous solution and let it stand for 7 days.

[0023] In an embodiment of the present invention, for economic reasons, the ethanol recovered from the filtrate during the vacuum concentration process in step (1) can be used to extract the filter residue twice more, and then the crude extract obtained from the three extractions can be combined for the next step of extraction.

[0024] Furthermore, the volume of water in step (2) is 9-11 L / kg (preferably 10 L / kg) based on the mass of the crude extract.

[0025] Furthermore, in step (2), the silica gel column chromatography is carried out using 200-300 mesh silica gel dry packing, and the mass ratio of the silica gel to the ethyl acetate extract is 1:1.

[0026] Furthermore, in step (5), the column used for the semi-preparative high-performance liquid chromatography is a Zorbax SB-C18 with a diameter of 9.4 mm, a length of 250 mm, and a particle size of 5 μm; the flow rate of the eluent is 1.5 mL / min.

[0027] The above elution process, including step (3), can be performed by TLC detection. Using chloroform:methanol at a volume ratio of 50:1 as the developing solvent, there is no absorption under ultraviolet light. The TLC is immersed in 10% sulfuric acid ethanol colorimetric reagent and heated to show a yellow-green color with an Rf value of 0.3, which is the point of target compound 2. Using chloroform:methanol at a volume ratio of 30:1 as the developing solvent, there is no absorption under ultraviolet light. The TLC is immersed in 10% sulfuric acid ethanol colorimetric reagent and heated to show a yellow-green color with an Rf value of 0.3, which is the point of target compound 1.

[0028] Thirdly, the present invention provides the application of the aforementioned erymophenol-type sesquiterpene compounds in the preparation of antitumor drugs.

[0029] Preferably, the erymophenol-type sesquiterpene compound is the compound shown in Formula 1.

[0030] In one embodiment of the present invention, the tumor is MCF-7 cells.

[0031] The novel sesquiterpenoid compounds involved in this invention can inhibit the proliferation of anti-MCF-7 cells and have good anti-tumor activity, and can be used to prepare anti-tumor drugs.

[0032] Compared with the prior art, the beneficial effects of this invention are as follows: This invention relates to two novel compounds 1 and 2, which have not been reported in existing anti-breast cancer drugs. Cell-level experiments have verified that these compounds exhibit good anti-proliferative activity against breast cancer cells and can be used to prepare anti-tumor compounds. Detailed Implementation

[0033] The present invention will be further described below with reference to specific embodiments, but the scope of protection of the present invention is not limited thereto:

[0034] Example 1: Isolation and extraction of new compounds 1 and 2

[0035] 10 kg of Lindera root was crushed and ground into powder, then sieved through an 80-mesh sieve. It was extracted by soaking in 50 L of 95% (v / v) ethanol (containing water) at room temperature (20–30°C) for seven days, then filtered. The ethanol in the filtrate was recovered under reduced pressure to obtain a crude extract. The recovered ethanol was used to soak the filter residue again. This extraction was repeated three times, and the crude extracts were combined to obtain a total of 1200 g. The crude extract was suspended in 13 L of water and extracted three times with 3 L of ethyl acetate. The ethyl acetate layer was concentrated under reduced pressure to obtain 265 g of ethyl acetate extract.

[0036] 256g of ethyl acetate extract was mixed with 256g of 200-300 mesh silica gel and packed into a 5120g column using a dry packing method with 200-300 mesh silica gel. The silica gel was eluted sequentially with a petroleum ether:ethyl acetate ratio of 15:1 → 1:1 (15:1, 10:1, 8:1, 6:1, 4:1, 2:1, 1:1), with each gradient eluting for 2-3 column volumes. The elution fractions at petroleum ether:ethyl acetate volumes of 4:1 and 2:1 were collected and concentrated under reduced pressure to obtain fractions A and B, respectively. During gradient elution, TLC detection can be performed. In segment A, using chloroform:methanol at a volume ratio of 50:1 as the developing solvent, there is no absorption under UV light. The TLC is immersed in 10% sulfuric acid ethanol colorimetric reagent, and the point where a yellow-green color is formed upon heating and the Rf value is 0.3 is the target compound 2. In segment B, using chloroform:methanol at a volume ratio of 30:1 as the developing solvent, there is no absorption under UV light. The TLC is immersed in 10% sulfuric acid ethanol colorimetric reagent, and the point where a yellow-green color is formed upon heating and the Rf value is 0.3 is the target compound 1.

[0037] Component B was concentrated under reduced pressure and eluted isocratically on an MCI column with a methanol-water ratio of 50:50. The developing conditions were chloroform:methanol ratio of 30:1. No color development was observed under UV light at 365 and 254 nm. The TLC fraction was then immersed in 10% sulfuric acid-ethanol reagent and heated to a yellow-green color with an Rf value of 0.3. The fractions were then combined and concentrated under reduced pressure. Subsequently, silica gel column chromatography was performed, eluting sequentially with petroleum ether:ethyl acetate gradients of 5:1, 4:1, 3:1, 2:1, and 1:1 (two column volumes for each gradient). The developing conditions were chloroform:methanol ratio of 30:1. No color development was observed under UV light at 365 and 254 nm. The TLC fraction was then immersed in 10% sulfuric acid-ethanol reagent and heated to a yellow-green color with an Rf value of 0.3. The fractions were then combined and concentrated under reduced pressure. 122 mg of the concentrate was dissolved in 1 mL of methanol and further eluted by ODS C-18 column chromatography with methanol:water volume ratios of 30:70, 40:60, 50:50, 60:60, 70:30, 80:20, 90:10, and 100:0, using chloroform:methanol volume ratio of 30:1 as the development condition. No color development was observed under UV light at 365 and 254 nm. Subsequently, the TLC fraction was immersed in 10% sulfuric acid ethanol colorimetric reagent and heated to produce a yellow-green color with an Rf value of 0.3. The fractions were combined and concentrated under reduced pressure to give compound 1 (34 mg).

[0038] Component A was eluted using an MCI column with gradients of methanol:water ratios of 30:70, 40:60, 50:50, 60:60, 70:30, 80:20, 90:10, and 100:0, eluting two column volumes for each gradient. The development conditions were chloroform:methanol at a ratio of 50:1. No color development was observed under UV light at 365 and 254 nm. Subsequently, TLC was performed by immersing the sample in 10% sulfuric acid-ethanol reagent and heating to develop a yellow-green color with an Rf value of 0.3. The fractions were then combined and concentrated under reduced pressure. Following this, silica gel chromatography was performed with isocratic elution using petroleum ether:ethyl acetate at a ratio of 10:1, with a chloroform:methanol ratio of 50:1. No color development was observed under UV light at 365 and 254 nm. The TLC was then performed by immersing the sample in 10% sulfuric acid-ethanol reagent and heating to develop a yellow-green color with an Rf value of 0.3. The fractions were then combined and concentrated under reduced pressure. The concentrate was dissolved in methanol and then eluted isocratically with an ODS C-18 column at a methanol:water ratio of 40:60, developing under chloroform:methanol ratio of 50:1. No color development was observed under UV light. The TLC fraction was then immersed in 10% sulfuric acid ethanol reagent and heated to a yellow-green color with an Rf value of 0.3. The fractions were then combined and concentrated under reduced pressure. Subsequently, the concentrate was eluted isocratically with a chloroform:methanol ratio of 100:1, developing under a chloroform:methanol ratio of 50:1 (v / v). No color development was observed under UV light. The TLC fraction was then immersed in 10% sulfuric acid ethanol reagent and heated to a yellow-green color with an Rf value of 0.3. The fractions were then combined and concentrated under reduced pressure. Finally, the concentrate was subjected to semi-preparative high-performance liquid chromatography (HPLC) with a methanol:water ratio of 79:21, and the t... R =22.3 min of the eluent was concentrated under reduced pressure to give compound 2 (16 mg).

[0039] Example 2: Structural identification of compounds 1-2

[0040] The structural formulas of the new compounds 1 and 2 were determined by mass spectrometry and nuclear magnetic resonance as follows:

[0041]

[0042] The physicochemical data of compound 1 are as follows: colorless monoclinic crystal, C 15 H 26 O2; (c 0.09,MeOH); 1HNMR(400MHz,CD3OD)δ5.31(s,1H),4.26(s,1H),2.43(dd,J=13.2,8.8Hz,1H),2.27–2.18(m,1H),1.99(dd,J=8.7,3.6Hz,2H),1.71(d,J= 13.7Hz,1H),1.54–1.50(m,2H),1.48(d,J=10.5Hz,2H),1.31(s,3H),1.19(s,3H),1.18–1.14(m,1H),0.97(s,3H),0.92(d,J=6.1Hz,3H). 13 C NMR (101MHz, CD3OD) δ142.3,123.1,73.9,71.8,46.4,41.2,39.5,35.6,30.9,28.6,28.5,28.1,26.1,21.0,16.4.; IR(ν max ):3328,2966,2920,1464,1435,1380,1276,1215,1146,1031,994,956,839,693cm -1 ;HR-ESI-MS:m / z 261.1827[M+Na] + (calcd.for C 15 H 26 NaO2 261.1825).

[0043]

[0044] The physicochemical data of compound 2 are as follows: colorless monoclinic crystals, C 17 H 28 O3; 63.6(c 0.11, MeOH); 1HNMR (400MHz, CDCl3) δ5.38 (d, J = 3.7Hz, 1H), 5.22–5.14 (m, 1H), 2.74 (dd, J=13.7,8.9Hz,1H),2.16–2.10(m,1H),2.09(d,J=1.3Hz,3H),1.99(d,J=7.6Hz,2H),1.81 –1.75(m,1H),1.53–1.49(m,2H),1.48–1.46(m,1H),1.47–1.43(m,1H),1.38–1.32(m,1H) ,1.21(d,J=1.4Hz,3H),1.18(d,J=1.4Hz,3H),0.96(d,J=1.4Hz,3H),0.93–0.89(m,3H).; 13C NMR (101MHz, CDCl3) δ170.4,139.8,123.9,75.1,72.0,45.0,38.3,37.4,34.6,30.9,28.5,28.4,26.9,25.3,21.7,20.7,16.1.; IR(ν max ):3423,1734,1463,1374,1245,1145,1025,963,834cm -1 ;HR-ESI-MS:m / z 303.1928[M+Na] + (calcd.for C 17 H 28 NaO3 303.1931).

[0045] Example 3: Test of antitumor activity of compounds

[0046] Experimental methods:

[0047] 1. Cell Culture: MCF-7 cells were cultured in Dalberg's essential medium (DMEM) containing 10% fetal bovine serum and 1% penicillin and streptomycin, in an incubator at 37°C and 5% CO2. When the cells reached the logarithmic growth phase, relevant experiments were performed.

[0048] 2. Cell viability assay: MCF-7 cells in logarithmic growth phase were prepared into a cell suspension and subjected to 5 × 10⁻⁶ cells / cells. 3 Add 100 μL of cell suspension to each well of a 96-well plate. After 24 h, treat the cells with 20 μM concentrations of compounds 1 and 2, respectively. Use 0.2 μM PTX (paclitaxel) as a control group. After culturing for 24 h, discard the culture medium. Dilute CCK-8 (Cell Counting Kit-8) and DMEM complete culture medium at a ratio of 1:10, and add 100 μL of diluent to each well. Incubate in the dark for another 2 h, and then measure the absorbance (OD) at 450 nm using a microplate reader.

[0049] Experimental results: The cell viability test results (Table 1) show that when MCF-7 cells were treated with 20 μM compound 1-2 for 24 h, the 20 μM compound 1 had a significant effect on cell viability.

[0050] Table 1. Inhibitory activity of compounds 1-2 on the proliferation of MCF-7 cells.

[0051]

Claims

1. A method for preparing an erymophenol-type sesquiterpene compound, characterized in that... The erymophenol-type sesquiterpenoid compound is one of the following: The preparation method is as follows: (1) Fragments of dried Lindera aggregata (Sims) Kosterm root were extracted with 95% ethanol aqueous solution, filtered, and the filtrate was concentrated under reduced pressure to obtain crude extract. (2) Disperse the crude extract obtained in step (1) evenly in water, extract with ethyl acetate, combine the ethyl acetate layers, concentrate under reduced pressure to obtain ethyl acetate extract; (3) The ethyl acetate extract obtained in step (2) was subjected to silica gel column chromatography. The silica gel column chromatography was performed by gradient elution with a mixed solvent of petroleum ether and ethyl acetate in volume ratios of 15:1, 10:1, 8:1, 6:1, 4:1, 2:1 and 1:1, respectively. Each gradient elution was performed for 2 to 3 column volumes. The elution fractions in volume ratios of 4:1 and 2:1 were collected and concentrated under reduced pressure to obtain component A and component B. (4) Take component B from step (3) and elute it with a mixed solvent of methanol and water in a volume ratio of 50:50 using an MCI column. Use a mixed solvent of chloroform and methanol in a volume ratio of 30:1 as the developing solvent for TLC detection. Combine the fractions with an Rf value of 0.3 and concentrate under reduced pressure to obtain concentrate B1. Concentrate B1 was subjected to silica gel column chromatography, with gradient elution using a mixed solvent of petroleum ether and ethyl acetate at volume ratios of 5:1, 4:1, 3:1, 2:1, and 1:1, two column volumes for each gradient. TLC was performed using a mixed solvent of chloroform and methanol at a volume ratio of 30:1 as the developing solvent. Fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate B2. The concentrate B2 was dissolved in methanol and subjected to ODS C-18 column chromatography. The ODS C-18 column chromatography was performed by gradient elution with mixed solvents of methanol and water in volume ratios of 30:70, 40:60, 50:50, 60:60, 70:30, 80:20, 90:10, and 100:0, with each gradient eluting two column volumes. The fractions were analyzed by TLC with a mixed solvent of chloroform and methanol in a volume ratio of 30:

1. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain the erymophenol-type sesquiterpene compound shown in Formula 1. (5) Take component A from step (3) and use an MCI column to perform gradient elution with a mixed solvent of methanol and water in volume ratios of 30:70, 40:60, 50:50, 60:60, 70:30, 80:20, 90:10, and 100:

0. Each gradient elution is two column volumes. Use a mixed solvent of chloroform and methanol in a volume ratio of 50:1 as the developing solvent for TLC detection. Combine the fractions with an Rf value of 0.3 and concentrate under reduced pressure to obtain concentrate A1. The concentrate A1 was subjected to silica gel column chromatography with a mixed solvent of petroleum ether and ethyl acetate in a volume ratio of 10:1 as the eluent and TLC detection with a mixed solvent of chloroform and methanol in a volume ratio of 50:1 as the developing solvent. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate A2. The concentrate A2 was dissolved in methanol and subjected to ODS C-18 column chromatography with a methanol-water mixture of 40:60 (v / v) as the eluent and a chloroform-methanol mixture of 50:1 (v / v) as the developing solvent. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate A3. The concentrate A3 was subjected to silica gel column chromatography using a mixed solvent of chloroform and methanol at a volume ratio of 100:1 as the eluent, and TLC was performed using a mixed solvent of chloroform and methanol at a volume ratio of 50:1 as the developing solvent. The fractions with an Rf value of 0.3 were combined and concentrated under reduced pressure to obtain concentrate A4. The concentrate A4 was eluted by semi-preparative high-performance liquid chromatography using a methanol-water mixture (79:21 v / v) as the eluent, and t was collected. R The eluent fraction of 22.3 min was concentrated under reduced pressure to obtain the erymophenol-type sesquiterpene compound shown in Formula 2.

2. The method for preparing the erymophenol-type sesquiterpene compound as described in claim 1, characterized in that: The volume of the ethanol-water solution in step (1) is 5 L / kg based on the mass of the Lindera root.

3. The method for preparing the erymophenol-type sesquiterpene compound as described in claim 1, characterized in that... The extraction process described in step (1) is as follows: the fragments of Lindera root are immersed in the ethanol aqueous solution at room temperature and left to stand for 7 days.

4. The method for preparing the erymophenol-type sesquiterpene compound as described in claim 1, characterized in that: The volume of water in step (2) is 9-11 L / kg based on the mass of the crude extract.

5. The method for preparing the erymophenol-type sesquiterpene compound as described in claim 1, characterized in that: In step (2), the silica gel column chromatography is performed using a 200-300 mesh silica gel dry packing method, and the mass ratio of the silica gel to the ethyl acetate extract is 1:

1.

6. The method for preparing the erymophenol-type sesquiterpene compound as described in claim 1, characterized in that: In step (5), the column used for the semi-preparative high-performance liquid chromatography is a Zorbax SB-C18 with a diameter of 9.4 mm, a length of 250 mm, and a particle size of 5 μm; the flow rate of the eluent is 1.5 mL / min.

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