A labdan-type diterpene glycoside extracted from aquilaria yunnanensis and use thereof

Abstract

The application provides a labdan-type diterpene glycoside extracted from Yunnan linaloe, and a use thereof, and belongs to the technical field of natural medicinal chemistry. The labdan-type diterpene glycoside is a compound shown in formula I or a salt thereof. The compound has good inhibitory activity on triple-negative breast cancer cells, and is expected to be used for preparing a drug for preventing and / or treating triple-negative breast cancer, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of natural medicinal chemistry technology, specifically relating to a labdanum-type diterpenoid glycoside extracted from Yunnan agarwood and its uses. Background Technology

[0002] Yunnan agarwood (Excoecaria acerifolia Didr.) is a species of agarwood belonging to the genus Excoecaria in the family Euphorbiaceae. It is mainly distributed at altitudes of 1200-3000 meters in Yunnan, Guizhou, Sichuan, and Hubei provinces of my country. Its core distribution area is in the Hengduan Mountains of Yunnan and Sichuan provinces, found in the hillsides and streamside thickets of shrubs and bamboo groves, with the Nujiang, Lancang, and Jinsha Rivers and their major tributaries as its main distribution areas. Among the ethnic minorities of Yunnan, including the Yi, Dai, Lahu, Tibetan, and Li, this plant is used to treat various ailments such as poisoning, inflammation, rheumatic pain, constipation, and colds.

[0003] Studies have shown that the traditional Chinese medicine *Aquilaria sinensis* from Yunnan contains various active ingredients such as terpenes and ketones. However, the specific compounds extracted from *Aquilaria sinensis* are currently few in number and their composition is unclear. Obtaining specific compound structures with antitumor activity from *Aquilaria sinensis* would be of great significance to the development of this traditional Chinese medicine.

[0004] Although labdanane-type diterpenoid glycosides are one type of diterpenoid, few chemical components possess antitumor activity. The labdanane-type diterpenoid compound ent-11α-hydroxy-3-oxo-13-epi-manoyl oxide, disclosed in the literature "Chemical Constituents, Bioactivity and Chemotaxonomy of Two Kinds of Daphne and Excoecaria acuminata [D]" (Huang Shengzhuo, Chinese Academy of Sciences, Kunming, 2012.), lacks activity. The labdanane-type diterpenoid compound ribenone, disclosed in the literature "Terpenoids and their anti-HIV-1 activities from Excoecaria acerifolia" (Huang SZ et al., Fitoterapia 2013, 91224-230.), also lacks antitumor activity. The labdane-type diterpenoid glycoside Hemsloside-Ma6, disclosed in the literature "Two new penterpenoid saponins and anew diterpenoid glycoside from Hemsleya chinensis" (Na-Li Song et al., Phytochemistry Letters, 2015, 13, 103-107.), showed no inhibitory activity against various cancer cells, including breast cancer cells MCF7. The compound ent-13-epi-8,13-epoxy-14S,15-dihydroxylabdan-3-one, disclosed in the literature "Five New Labdane-Type Diterpenes from Excoecaria agallocha.IV" (Konishi T. et al., Chemical & Pharmaceutical Bulletin, 1998, 46(9):1393-1398.), also did not exhibit cytotoxic activity. Therefore, developing a labdane-type diterpenoid glycoside with antitumor activity is of great significance. Summary of the Invention

[0005] The purpose of this invention is to provide a labdanane-type diterpenoid glycoside extracted from Yunnan agarwood and its uses.

[0006] This invention provides compounds of Formula I or salts thereof:

[0007]

[0008] Furthermore, the compound is as shown in Formula II:

[0009]

[0010] The present invention also provides a method for preparing the aforementioned compound, comprising the following steps:

[0011] (1) Extraction of extract: Take Yunnan agarwood powder, reflux extract with 85-95% methanol, remove solvent by vacuum distillation of the extract to obtain crude methanol extract;

[0012] (2) Organic solvent extraction: The crude methanol extract was suspended in water and extracted sequentially with ethyl acetate and n-butanol. The solvent was removed by vacuum distillation to obtain the n-butanol extract fraction.

[0013] (3) First normal-phase silica gel column chromatography: The n-butanol extract was dissolved in a mixed solvent of dichloromethane and methanol. The dissolved sample was adsorbed onto silica gel and dry-packed. Gradient elution was performed with mixed solutions of dichloromethane and methanol at volume ratios of 1:0, 50:1, 20:1, 10:1, and 1:1. After removing the solvent by vacuum distillation, the corresponding eluted samples Fr.1, Fr.2, Fr.3, Fr.4, and Fr.5 were obtained. Fr.3 was dissolved in methanol and adsorbed onto an RP-18 reversed-phase silica gel column. Dry reversed-phase column chromatography was performed. Elution was performed sequentially with methanol aqueous solutions at concentrations of 40%, 60%, and 85%. After removing the solvent by vacuum distillation, the corresponding eluted samples Fr.3a, Fr.3b, and Fr.3c were obtained.

[0014] (4) Second normal phase silica gel column chromatography: The sample Fr.3b was dissolved and eluted using a mixed solvent of dichloromethane and methanol, adsorbed onto silica gel, and dry-packed into a column. Isocratic elution was performed using a mixed solvent of petroleum ether and ethyl acetate with a volume ratio of 3:1. The sulfuric acid color was detected by developing a purple-red spot with a volume ratio of petroleum ether to ethyl acetate of 2:1. The Rf value was 0.3 to 0.5. The fractions were combined and the solvent was removed under reduced pressure to obtain the crude compound.

[0015] (5) Gel chromatography purification: The crude compound was dissolved in methanol and subjected to dextran gel chromatography. Methanol was used as the eluent and petroleum ether and ethyl acetate were used as the developing solvent in a volume ratio of 2:1 to detect the purple-red spots of sulfuric acid. The Rf value was 0.3 to 0.5. The distillate of the trapezoidal spots was taken and the solvent was removed under reduced pressure to obtain the compound.

[0016] Furthermore,

[0017] In step (1), the reflux extraction is performed 1 to 3 times, and the mass-to-volume ratio of Yunnan agarwood powder to methanol is 1 kg: 5 to 10 L during each reflux extraction. The reflux extraction time is 1 to 5 hours and the temperature is 60 to 80 °C.

[0018] And / or, in step (3), the volume ratio of dichloromethane to methanol in the dichloromethane and methanol mixed solvent is 1:1;

[0019] And / or, in step (3), the silicone is 200-300 mesh silicone;

[0020] And / or, in step (3), the volume-to-mass ratio of the dichloromethane and methanol mixture solution with a volume ratio of 1:0, 50:1, 20:1, 10:1, and 1:1 to the n-butanol extract is 50L:355.0g, 50L:355.0g, 50L:355.0g, 50L:355.0g, and 20L:355.0g, respectively;

[0021] And / or, in step (3), the volume-to-mass ratio of the methanol aqueous solution with concentrations of 40%, 60%, and 85% to Fr.3 is 8.0L:13.0g, 8.0L:13.0g, and 5.0L:13.0g, respectively;

[0022] And / or, in step (4), the volume ratio of dichloromethane to methanol in the dichloromethane and methanol mixed solvent is 10:1;

[0023] And / or, in step (4), the silicone is 300-400 mesh silicone.

[0024] Furthermore,

[0025] In step (1), the reflux extraction is performed 3 times. The mass-to-volume ratio of Yunnan agarwood powder to methanol is 1 kg: 5 L during each reflux extraction. The reflux extraction time is 4 hours and the temperature is 70℃.

[0026] The present invention also provides the use of the aforementioned compounds or salts thereof in the preparation of medicaments for the prevention and / or treatment of cancer.

[0027] Furthermore, the cancer in question is breast cancer.

[0028] Furthermore, the breast cancer mentioned is triple-negative breast cancer.

[0029] The present invention also provides a medicament for the prevention and / or treatment of cancer, which is prepared by using the aforementioned compound or its salt as the active ingredient, plus pharmaceutically acceptable excipients or auxiliary ingredients.

[0030] Furthermore, the cancer is breast cancer; preferably, the cancer is triple-negative breast cancer.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] This invention extracts a labdanum-type diterpenoid glycoside compound from Yunnan agarwood. This compound has good inhibitory activity against triple-negative breast cancer cells and is expected to be used in the preparation of drugs for the prevention and / or treatment of triple-negative breast cancer, showing promising application prospects.

[0033] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions, or alterations can be made without departing from the basic technical concept of the present invention.

[0034] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0035] Figure 1 The nuclear magnetic resonance hydrogen spectrum of the labdanane-type diterpenoid glycosides of the present invention ( 1 H NMR spectrum).

[0036] Figure 2 The carbon NMR spectrum of the labdanane-type diterpenoid glycosides of this invention ( 13 (C NMR and DEPT spectra). Detailed Implementation

[0037] Where specific techniques or conditions are not specified in the detailed embodiments of this invention, they shall be performed in accordance with the techniques or conditions described in the literature in this field or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased.

[0038] Unless otherwise stated, the proportions in this invention are volume ratios and the concentrations are volume percentage concentrations.

[0039] Example 1: Preparation and Identification of Laudanane-type Diterpenoid Glycosides of the Present Invention

[0040] 1. Preparation of the labdanane-type diterpenoid glycoside of the present invention

[0041] (1) Extraction of extract: Take 18.0 kg of dried Yunnan agarwood powder, crush it and extract it three times by heating and reflux with 90% methanol (MeOH) (70℃; extraction time for each extraction is 4 hours; the mass-volume ratio of Yunnan agarwood powder to 90% methanol is 1 kg: 5 L each time). Concentrate the extract under reduced pressure to remove the solvent and combine the extract to obtain crude methanol extract.

[0042] (2) Organic solvent extraction: The crude methanol extract was suspended in water (15.5L) and extracted sequentially with ethyl acetate and n-butanol (15L×3). The solvent was removed by vacuum concentration to obtain 355.0g of n-butanol extract.

[0043] (3) First normal-phase silica gel column chromatography: 355.0 g of n-butanol extract was dissolved in a mixed solvent of dichloromethane and methanol (volume ratio 1:1) (4.0 L), and the sample was adsorbed onto 200-300 mesh silica gel (0.80 kg); the column was dry packed, and gradient elution was performed with dichloromethane and methanol (volume ratios of 1:0; 50:1; 20:1; 10:1; 1:1, with volumes of 50 L, 50 L, 50 L, 50 L, and 20 L, respectively). After removing the solvent by vacuum distillation, the corresponding eluted samples Fr.1 (56.0 g), Fr.2 (7.0 g), Fr.3 (13.0 g), Fr.4 (76.0 g), and Fr.5 (152.5 g) were obtained. Sample Fr.3 (13.0 g) was dissolved in methanol and adsorbed onto an RP-18 reversed-phase silica gel column. Dry reversed-phase column chromatography was performed, with elution successively with 40% methanol (8.0 L), 60% methanol (8.0 L), and 85% methanol (5.0 L). After concentration under reduced pressure to remove the solvent, corresponding eluted samples Fr.3a (3.6 g), Fr.3b (4.0 g), and Fr.3c (2.0 g) were obtained.

[0044] (4) Second normal-phase silica gel column chromatography separation: The sample Fr.3b was dissolved and eluted with a mixed solvent of dichloromethane and methanol in a volume ratio of 10:1, and then adsorbed onto silica gel of 300-400 mesh and dry-packed; isocratic elution was performed with a mixed solvent of petroleum ether and ethyl acetate in a volume ratio of 3:1, and the sulfuric acid color development was detected with a developing solvent of petroleum ether and ethyl acetate in a volume ratio of 2:1, resulting in purple-red spots (Rf value 0.3-0.5). The fractions were combined and the solvent was removed under reduced pressure to obtain the crude labdanum-type diterpenoid glycoside.

[0045] (5) Gel chromatography purification: Dissolve the crude labdanane-type diterpenoid glycoside in methanol, take 2 mL to 3 mL of methanol to make the mass-volume ratio of crude product to methanol 1 g: 1 mL, and perform dextran gel chromatography. Use methanol as eluent and petroleum ether to ethyl acetate as the developing solvent in a volume ratio of 2:1 to detect the purple-red spots (Rf value 0.3 to 0.5) that are developed by sulfuric acid. Take the distillate of the trapezoidal spots, remove the solvent under reduced pressure, and obtain the pure labdanane-type diterpenoid glycoside (8.2 mg).

[0046] The structure of the pure labdanane-type diterpenoid glycosides obtained by the above method was determined by the following method.

[0047] The obtained pure labdanane-type diterpenoid glycosides were white amorphous powders. High-resolution Fourier transform mass spectrometry (HR-FT-MS) of the pure labdanane-type diterpenoid glycosides showed a quasi-molecular ion peak at m / z 523.2874 [M+Na]. + (C 26 H 44 O9Na + [M+Na]+ The calcd.for 523.2877 indicates that the molecular formula of this compound is C. 26 H 44 O9, with an unsaturation degree of 5. Infrared spectrum ν max 3440cm -1 and 1702cm -1 This indicates that the compound contains hydroxyl and carbonyl groups. 1 Characteristic signals in H-NMR spectra (see Table 1): δ H 5.00 (d, J = 7.7 Hz, 1H), δ H The values ​​of 4.54 (d, J = 10.3 Hz, 1H) and 4.37 (dd, J = 11.6, 5.3 Hz, 1H) suggest that the compound contains a β-glucose structural fragment. GC analysis of the monosaccharide obtained from acid hydrolysis of this compound showed a retention time consistent with that of D-Glc (Rt = 23.18 min), confirming that the sugar fragment of this compound is D-glucanopyranose. 1 The characteristic signal of five angular methyl groups in the H-NMR spectrum: δ H 1.27(s), 1.34(s), 1.09(s), 0.98(s), 0.75(s). 13 C-NMR and DEPT spectra contain glycosyl signals δ C Aglycone signals at 105.8(d), 76.4(d), 78.8(d), 72.0(d), 78.8(d), 63.1(t) and 20 carbons, including 5 methyl groups: δ C 26.2(q,C-16), 24.4(q,C-17), 26.8(q,C-18), 21.4(q,C-19), 15.1(q,C-20), 7 methylene groups: δ C 38.1(t,C-1), 34.3(t,C-2), 21.2(t,C-6), 43.5(t,C-7), 15.7(t,C-11), 31.5(t,C-12), 73.0(t,C-15), 3 methines: δ C 55.0 (d, C-5), 54.9 (d, C-9), 75.7 (d, C-14) and 5 quaternary carbons: δ C216.3(s,C-3), 47.6(s,C-4), 75.2(s,C-8), 36.9(s,C-10), 75.2(s,C-13), these information suggest that the compound has a labdane-type diterpenoid skeleton. A careful comparison of the NMR data of this compound with that of the known compound ent-13-epi-8,13-epoxy-14S,15-dihydroxylabdan-3-one (prepared according to the literature Konishi T., Fujiwara Y., Konoshima T., Kiyosawa S. Five New Labdane-Type Diterpenes from Excoecaria agallocha.IV[J]. Chemical & Pharmaceutical Bulletin, 1998, 46(9):1393-1398., structure shown in the figure below), reveals that the NMR spectrum of this compound shows an additional signal of a β-glucose structural fragment, and the chemical shift at position C-15 is due to δ C 63.5 shifts to lower fields to δ C 73.0 indicates that the compound is a glycoside of ent-13-epi-8,13-epoxy-14S,15-dihydroxylabdan-3-one.

[0048]

[0049] The structural formula of ent-13-epi-8,13-epoxy-14S,15-dihydroxylabdan-3-one

[0050] In the HMBC spectrum, δ H 5.00 (d, J = 7.7 Hz, H⁻¹') and δ C A significant correlation exists between 73.0 (t, C-15), further demonstrating that the β-glucose structural fragment is linked to the C-15 position of the aglycone. Additionally, in the HMBC spectrum, δ H 1.09 (3H, s, C-18), δ H 0.98 (3H, s, C-18) and δ C 216.3 shows a significant correlation. In the NOESY plot, δ H 4.09 (m, H-14) and δ HThe presence of NOE in 1.27(s,H-16) suggests that the configuration of OH-14 is -α. Therefore, the compound obtained in this invention is identified as ent-13-epi-8,13-epoxy-14(α),15-dihydroxylabdan-3-one-15-O-β-D-glucopyranoside, with the structure shown below:

[0051]

[0052] The structural formula of the compound obtained in this invention

[0053] Table 1. NMR data (δinppm, Jin Hz) of the compounds of this invention and known compounds

[0054]

[0055]

[0056]

[0057] The following specific experimental examples demonstrate the beneficial effects of the present invention.

[0058] Experimental Example 1: The inhibitory activity of the compound ent-13-epi-8,13-epoxy-14(α),15-dihydroxylabdan-3-one-15-O-β-D-glucopyranoside, prepared in Example 1 of the present invention, against tumor cells.

[0059] 1. Detection Principle

[0060] The antitumor activity of the compounds prepared in this invention was determined using the MTT assay. The MTT assay involves the reduction of exogenous thiazolyl blue (MTT) into blue-violet crystals by succinate dehydrogenase in the mitochondria of living cells; this process differs from the reaction in dead cells. Subsequently, formazan was dissolved in cell-grade dimethyl sulfoxide, mixed thoroughly, and its absorbance (OD) value was measured at 570 nm using a microplate reader. Within a certain cell number range, the amount of MTT crystals formed was directly proportional to the cell number. The antitumor cell activity was calculated based on the OD value.

[0061] The following cell lines were used in this part of the experiment: triple-negative breast cancer (TNBC) cell line (HCC1806), human B lymphocyte cell line (ST486), colon cancer cell line (CT26), human cervical cancer cell line (HeLa), and human lung cancer cell line (A549).

[0062] 2. Testing Steps

[0063] 1) Culture cells in RPMI 1640 medium containing 10% fetal bovine serum. Based on the cell line's growth characteristics, take cells in the logarithmic growth phase and culture them at 5.0 × 10⁻⁶ cells / mL. 3 Add the required number of cells to the culture plate at 100 μL / well, and spread them evenly on a 96-well culture plate. To avoid edge effects, fill the edge wells with an equal volume of PBS.

[0064] 2) Cells were incubated at 37°C with 5% CO2 for 24 hours. Compounds at concentrations of 2.5, 5, 10, 20, 40, and 80 μM were then used after dissolution in DMSO. 100 μL of each concentration was added to each well as a blank control. Three replicates were set up for each concentration, and the average value was calculated.

[0065] 3) After incubating in an incubator at 37°C with 5% CO2 for 48 hours, observe cell growth. Weigh MTT and prepare a 5 mg / mL solution with PBS, then repeatedly shake to mix.

[0066] 4) Add 20 μL of MTT solution per well, and incubate at 37°C for 4 hours under 5% CO2 conditions.

[0067] 5) Gently remove the supernatant culture medium from each well until completely dissolved, add 150 μL of DMSO per well, and shake evenly for a few seconds with a microplate shaker to ensure that MTT-formazan is dissolved evenly.

[0068] 6) Turn on the microplate reader and measure the absorbance at 570 nm.

[0069] 7) Use Origin software (www.originlab.com) to calculate the half-maximal inhibitory concentration (IC50) for each compound. 50 , half-maximal inhibitory concentration).

[0070] 3. Results

[0071] The inhibitory activities of the compounds of this invention on various tumor cells are shown in Table 2.

[0072] Table 2. Inhibitory activity of the compounds of the present invention against various tumor cells

[0073]

[0074] a IC 50 Data This indicates that repeated testing was performed using 3 wells each time.

[0075] Table 2 shows that: The literature "Five New Labdane-Type Diterpenes from Excoecaria agallocha.IV" (Konishi T. et al., Chemical & Pharmaceutical Bulletin, 1998, 46)

[0076] (9):1393-1398.) The known labdanane-type diterpenoid compound ent-13-epi-8,13-epoxy-14S,15-dihydroxylabdan-3-one, and the compound ent-11α- in the literature "Chemical Constituents, Bioactivity and Chemotaxonomy of Two Kinds of Daphne and Scutellaria barbata [D]" (Huang Shengzhuo. Chinese Academy of Sciences, Kunming, 2012.)

[0077] Neither hydroxy-3-oxo-13-13-epi-manoyl oxide nor ent-11α-hydroxy-3-oxo-13-epi-manoyl oxide, nor the compound ribenone mentioned in the literature "Terpenoids and their anti-HIV-1 activities from Excoecaria acerifolia" (Huang SZ et al., Fiteterapia 2013, 91224-230.) showed any inhibitory activity against tumor cells.

[0078] In addition, the literature "Two new penterpenoid saponins and a new diterpenoid glycoside from Hemsleya chinensis" (Na-Li Song et al., Phytochemistry Letters)

[0079] The compound Hemsloside-Ma6, described in 2015, 13, 103-107, showed no inhibitory activity against any of the following cell lines: breast cancer cell line (MCF7), liver cancer cell line (HepG2), colon cancer cell line (HT29), and gastric cancer cell line (MKN28).

[0080] The compound of this invention showed good inhibitory activity against triple-negative breast cancer cells (HCC1806), IC50. 50 The value was (61.91±7.99) μM.

[0081] In summary, this invention extracts a labdanum-type diterpenoid glycoside compound from Yunnan agarwood. This compound exhibits good inhibitory activity against triple-negative breast cancer cells and is expected to be used in the preparation of drugs for the prevention and / or treatment of triple-negative breast cancer, showing promising application prospects.

Claims

1. A compound or a salt thereof, characterized in that: The compound is shown in Formula II: Formula II.

2. A method for preparing the compound of claim 1, characterized in that: It includes the following steps: (1) Extraction of extract: Take Yunnan agarwood powder, reflux extract with 85-95% methanol, remove solvent by vacuum distillation of extract to obtain crude methanol extract; (2) Organic solvent extraction: The crude methanol extract was suspended in water and extracted with ethyl acetate and n-butanol in sequence. The solvent was removed by vacuum distillation to obtain the n-butanol extract fraction. (3) First normal-phase silica gel column chromatography: The n-butanol extract was dissolved in a mixed solvent of dichloromethane and methanol. The dissolved sample was adsorbed onto silica gel and dry-packed. Gradient elution was performed with mixed solutions of dichloromethane and methanol at volume ratios of 1:0, 50:1, 20:1, 10:1, and 1:

1. After removing the solvent by vacuum distillation, the corresponding eluted samples Fr.1, Fr.2, Fr.3, Fr.4, and Fr.5 were obtained. Fr.3 was dissolved in methanol and adsorbed onto an RP-18 reversed-phase silica gel column. Dry reversed-phase column chromatography was performed. Elution was performed sequentially with methanol aqueous solutions at concentrations of 40%, 60%, and 85%. After removing the solvent by vacuum distillation, the corresponding eluted samples Fr.3a, Fr.3b, and Fr.3c were obtained. (4) Second normal phase silica gel column chromatography: The sample Fr.3b was dissolved and eluted using a mixed solvent of dichloromethane and methanol, adsorbed onto silica gel, and dry-packed into a column. Isocratic elution was performed using a mixed solvent of petroleum ether and ethyl acetate with a volume ratio of 3:

1. The sulfuric acid color was detected by developing a purple-red spot with a volume ratio of petroleum ether to ethyl acetate of 2:

1. The Rf value was 0.3~0.

5. The fractions were combined and the solvent was removed under reduced pressure to obtain the crude compound. (5) Gel chromatography purification: The crude compound was dissolved in methanol and subjected to dextran gel chromatography. Methanol was used as the eluent and petroleum ether and ethyl acetate were used as the developing solvent in a volume ratio of 2:1 to detect the purple-red spots of sulfuric acid. The Rf value was 0.3~0.

5. The distillate of the trapezoidal spots was taken and the solvent was removed under reduced pressure to obtain the compound.

3. The method according to claim 2, characterized in that: In step (1), the reflux extraction is performed 1 to 3 times, and the mass-to-volume ratio of Yunnan agarwood powder to methanol is 1 kg: 5 to 10 L during each reflux extraction. The reflux extraction time is 1 to 5 hours and the temperature is 60 to 80 °C. In step (3), the volume ratio of dichloromethane to methanol in the dichloromethane and methanol mixed solvent is 1:1; In step (3), the silicone is 200-300 mesh silicone; In step (3), the volume-to-mass ratios of the dichloromethane and methanol mixed solutions with volume ratios of 1:0, 50:1, 20:1, 10:1, and 1:1 to the n-butanol extract are 50 L: 355.0 g, 50 L: 355.0 g, 50 L: 355.0 g, 50 L: 355.0 g, and 20 L: 355.0 g, respectively. In step (3), the volume-to-mass ratios of the 40%, 60%, and 85% methanol aqueous solutions and Fr.3 are 8.0 L:13.0 g, 8.0 L:13.0 g, and 5.0 L:13.0 g, respectively. In step (4), the volume ratio of dichloromethane to methanol in the dichloromethane and methanol mixed solvent is 10:1; In step (4), the silicone is 300~400 mesh silicone.

4. The method according to claim 3, characterized in that: In step (1), the reflux extraction is performed 3 times. The mass-to-volume ratio of Yunnan agarwood powder to methanol is 1 kg: 5 L during each reflux extraction. The reflux extraction time is 4 hours and the temperature is 70 °C.

5. Use of the compound of claim 1 or a salt thereof in the preparation of a medicament for the prevention and / or treatment of cancer, wherein the cancer is triple-negative breast cancer.

Images