A sesquiterpene dimer compound, preparation method and application

Abstract

The application provides a sesquiterpene dimer compound, a preparation method and application, and belongs to the field of medical biotechnology. The sesquiterpene dimer compound is extracted and separated from a Lemnalia soft coral to obtain a sesquiterpene dimer compound lemnalinoid L. The sesquiterpene dimer compound lemnalinoid L has an antitumor effect, has inhibitory activity on a human breast cancer cell line MDA-MB-231, can be used for developing a candidate molecule for treating the disease, and can be used as a lead compound for chemical synthesis and structural modification of other drugs. The preparation method is simple and fast, and the extracted new diterpene glycoside compound has high purity.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a sesquiterpene dimer compound, its preparation method, and its application. Background Technology

[0002] Sesquiterpenes are the largest class of natural terpenes, with a rich variety of structures, including more than 5,000 types and more than 100 skeleton types. They have a variety of biological activities and are found not only in medicinal plants but also in marine organisms.

[0003] Current research has found that the main secondary metabolites of soft corals in the genus *Lemnalia* are sesquiterpenes, including sesquiterpenes with various carbon skeletons, such as nardosinane, neolemnane, and ylangane sesquiterpenes. Many sesquiterpenes with antitumor, antibacterial, antiviral, and neuroprotective properties have been discovered. Compared to monomeric compounds, dimers exhibit more significant and unique biological activities; however, the discovery of sesquiterpene dimers is extremely limited. Summary of the Invention

[0004] Based on the above-mentioned technical problems to be solved, the present invention provides a sesquiterpene dimer compound, a preparation method thereof, and its application.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: In one aspect, this application provides a sesquiterpene dimer compound extracted from soft corals of the genus *Lemnalia*. The sesquiterpene dimer compound in this application is named lemnalinoid L, is a white powder, and has the molecular formula C2. 32 H 45 NO6, chemical structural formula is: .

[0006] Secondly, this application provides a method for preparing a sesquiterpene dimer compound, the method comprising: S01: Soft coral is cut into pieces and soaked in methanol at room temperature to obtain an extract. The extract is then concentrated under reduced pressure, desalted with anhydrous methanol, and concentrated under reduced pressure again to obtain a crude extract.

[0007] Frozen Lemnalia soft corals were thawed at room temperature and cut into small pieces about the size of a fingernail. The cut corals were then soaked in room temperature methanol six times, for three days each time. After each soaking, the solutions were filtered and combined to obtain an extract. The extract was concentrated under reduced pressure at 37°C and 0.1 MPa to a paste-like consistency. Anhydrous methanol was added to the concentrated extract to redissolve it. After filtration and desalting, the extract was concentrated under reduced pressure at 37°C and 0.1 MPa to obtain a crude extract.

[0008] S02: The crude extract was separated by silica gel vacuum column chromatography, gradient elution, thin-layer chromatography colorimetric detection, and the same fractions were combined and concentrated to obtain 16 primary components F1-F16; wherein, the gradient elution was performed sequentially by petroleum ether, petroleum ether / acetone with volume ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, dichloromethane, and dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1.

[0009] The crude extract was separated by chromatographic chromatography using a 160mm × 500mm silica gel column with a particle size of 200-300 mesh under reduced pressure. The separated products were eluted sequentially with petroleum ether, petroleum ether / acetone (v / v ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1), dichloromethane, and dichloromethane / methanol (v / v ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1) to obtain the eluent fractions. Thin-layer chromatography was performed using petroleum ether and propanol (v / v ratio of 3:1) as the developing solvent and 10% sulfuric acid in ethanol as the colorimetric reagent. Fractions with the same volume were combined and concentrated to obtain 16 primary fractions F1-F16.

[0010] S03: F12 in the primary components F1-F16 was separated by silica gel column chromatography, gradient elution, thin-layer chromatography colorimetric detection, and the same fractions were combined and concentrated to obtain four secondary components F121-F124; wherein, gradient elution was performed sequentially using petroleum ether / acetone in volume ratios of 20 / 1, 10 / 1, 5 / 1, and 0 / 1.

[0011] Component F12 from the primary fractions F1-F16 was separated chromatographically using a 60mm × 500mm silica gel column with 300-400 mesh particles. The separated products were sequentially eluted with petroleum ether / acetone at volume ratios of 20 / 1, 10 / 1, 5 / 1, and 0 / 1 to obtain the eluent fractions. Thin-layer chromatography was performed on the eluent fractions using petroleum ether and propanol at a volume ratio of 3:1 as the developing solvent and 10% sulfuric acid in ethanol as the colorimetric reagent. Fractions with the same distillate were combined and concentrated to obtain four secondary fractions F121-F124.

[0012] S04: After high performance liquid chromatography analysis, medium-pressure preparative liquid chromatography separation, and isocratic elution, tertiary components F124Q, F124F, and F124H are obtained from the secondary components F121-F124; wherein, isocratic elution is performed using methanol / water with a volume ratio of 80 / 20.

[0013] High-performance liquid chromatography (HPLC) and medium-pressure preparative liquid chromatography (PCLC) were used to analyze and separate F124 in the secondary fraction F121-F124, yielding the separated product. The HPLC analysis conditions were as follows: an analytical column of 4.6 mm × 250 mm with 5 μm particle size from *Lysimachia christinae* ODS C20. 18 The column was prepared at a flow rate of 1 ml / min with a methanol / water volume ratio of 70 / 30. The medium-pressure preparative liquid chromatography separation conditions were as follows: a 20 mm × 250 mm preparative column with a particle size of 5 μm, containing *Lysimachia christinae* ODS C. 18 The column was used at a flow rate of 10 ml / min.

[0014] The separated products were isocratically eluted with a methanol / water ratio of 70 / 30 (v / v) to obtain three tertiary fractions: F124Q, F124F, and F124H. Based on the retention times in the high-performance liquid chromatograms of fraction F124, the fractions were determined as follows: 0.00–3.50 min for F124Q; 3.50–7.60 min for F124F; and 7.60–9.50 min for F124H.

[0015] S05: The tertiary component F124H was analyzed by high performance liquid chromatography, separated by high performance preparative liquid chromatography, and eluted isocratically to obtain a sesquiterpene dimer compound; wherein isocratically eluted with acetonitrile / water at a volume ratio of 40 / 60.

[0016] The tertiary component F124H was purified and analyzed by high-performance liquid chromatography (HPLC). The separation and preparative chromatographic conditions were determined to be acetonitrile / water with a volume ratio of 40 / 60. The HPLC analysis conditions were as follows: an analytical column of 4.6 mm × 250 mm with a particle size of 5 μm, containing *Lysimachia christinae* ODS C. 18The column was used with a flow rate of 1 ml / min. High-performance preparative liquid chromatography (HPLC) was employed to separate and purify component F124H. The separated product was isocratically eluted with acetonitrile / water at a volume ratio of 40 / 60 at a retention time of 156 min to obtain the sesquiterpene dimer compound lemnalinoid L. The HPLC separation conditions were as follows: a 10 mm × 250 mm column with a particle size of 5 μm; and a *Lysimachia christinae* ODS C20 column. 18 The column was used at a flow rate of 2 ml / min.

[0017] Thirdly, this application provides an application of the sesquiterpene dimer compound lemnalinoid L, namely, its application in the preparation of drugs that inhibit human breast cancer cell lines.

[0018] The present invention has the following beneficial effects: (1) This application is the first to extract and isolate the sesquiterpene dimer compound lemnalinoid L from soft corals of the genus Lemnalia. The molecular structure, isolation method and pharmacological activity of lemnalinoid L have not been reported in existing journal articles.

[0019] (2) The sesquiterpene dimer compound lemnalinoid L has antitumor activity and inhibits human breast cancer cell line MDA-MB-231. It can be used to develop candidate molecules for the treatment of this disease, and can also be used as a lead compound for the chemical synthesis and structural modification of other drugs.

[0020] (3) The extraction method of the sesquiterpene dimer compound lemnalinoid L is simple and fast, and the extracted compound has high purity. Attached Figure Description

[0021] Figure 1 The UV (Ultraviolet Visible Absorption Spectroscopy) spectrum of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application is shown below. Figure 2 The IR (Infrared Spectroscopy) spectrum of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application is shown below. Figure 3 The image shows the HRESIMS (high-resolution electrospray ionization mass spectroscopy) spectrum of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application. Figure 4The sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application 1 1H-NMR (Nuclear Magnetic Resonance Spectroscopy of Hydrogen) spectrum; Figure 5 The sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application 13 C-NMR (Carbon-13 nuclear magnetic resonance) spectrum; Figure 6 The HSQC (heteronuclear singular quantum correlation) spectrum of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application is shown below. Figure 7 The HMBC (1H-detected heteronuclear multiple bond correlation) spectrum of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application is shown below. Figure 8 The image shows the 1H-1H COSY (Homonuclear Chemical Shift Correlation Spectroscopy) spectrum of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application. Figure 9 The NOSEY (Nuclear Overhauser Effect Spectroscopy) diagram of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application is shown below. Figure 10 This is a schematic diagram of the HMBC and 1H-1HCOSY of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application; Figure 11 This is a comparison chart of the TDDFT-ECD calculation and experimental CD value of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application. Detailed Implementation

[0022] The technical solution of the present invention will be further explained and described below through specific embodiments.

[0023] Example 1 This application provides a sesquiterpene dimer compound extracted from soft corals of the genus *Lemnalia*. The sesquiterpene dimer compound in this application is named lemnalinoid L, is a white powder, and has the molecular formula C2. 32 H 45 NO6, chemical structural formula is: .

[0024] Example 2 This application provides a method for preparing a sesquiterpene dimer compound, comprising: S201: Thaw frozen Lemnalia soft corals at room temperature and cut them into small pieces about the size of a fingernail. Soak the cut corals in room temperature methanol six times, for three days each time. After soaking, filter and combine all the soaking solutions to obtain the extract. Concentrate the extract under reduced pressure at 37°C and 0.1 MPa to a paste. Add anhydrous methanol to the concentrated extract to redissolve it, filter to desalinate, and then concentrate under reduced pressure at 37°C and 0.1 MPa to obtain a crude extract.

[0025] S202: The crude extract was separated by chromatographic chromatography using a 160mm×500mm silica gel column with a silica gel particle size of 200-300 mesh under reduced pressure. The separated products were eluted sequentially with petroleum ether, petroleum ether / acetone (v / v ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1), dichloromethane, and dichloromethane / methanol (v / v ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1) to obtain the eluent fractions. Thin-layer chromatography was performed using petroleum ether and propanol (v / v ratio of 3:1) as the developing solvent and 10% sulfuric acid in ethanol as the colorimetric reagent. Fractions with the same distillate were combined and concentrated to obtain 16 primary fractions F1-F16.

[0026] S203: Component F12 from the primary fractions F1-F16 was separated chromatographically using a 60mm×500mm silica gel column with 300-400 mesh particles. The separated products were eluted sequentially with petroleum ether / acetone at volume ratios of 20 / 1, 10 / 1, 5 / 1, and 0 / 1 to obtain the eluent fractions. Thin-layer chromatography was performed on the eluent fractions using petroleum ether and propanol at a volume ratio of 3:1 as the developing solvent and 10% sulfuric acid in ethanol as the colorimetric reagent. Components with the same fraction were combined and concentrated to obtain four secondary fractions F121-F124.

[0027] S204: High-performance liquid chromatography (HPLC) and medium-pressure preparative liquid chromatography (PCLC) were used sequentially to analyze and separate F124 from the secondary fraction F121-F124, yielding the separated product. The HPLC analysis conditions were as follows: an analytical column of 4.6 mm × 250 mm with 5 μm particle size, containing *Lysimachia christinae* ODS C. 18 The column was prepared at a flow rate of 1 ml / min with a methanol / water volume ratio of 70 / 30. The medium-pressure preparative liquid chromatography separation conditions were as follows: a 20 mm × 250 mm preparative column with a particle size of 5 μm, containing *Lysimachia christinae* ODS C. 18 The column was used at a flow rate of 10 ml / min.

[0028] The separated products were isocratically eluted with a methanol / water ratio of 70 / 30 (v / v) to obtain three tertiary fractions: F124Q, F124F, and F124H. Based on the retention times in the high-performance liquid chromatograms of fraction F124, the fractions were determined as follows: 0.00–3.50 min for F124Q; 3.50–7.60 min for F124F; and 7.60–9.50 min for F124H.

[0029] S205: The tertiary component F124H was purified and separated by high-performance liquid chromatography (HPLC). The HPLC conditions were determined to be an acetonitrile / water mixture with a volume ratio of 40 / 60. The HPLC analysis conditions were as follows: an analytical column of 4.6 mm × 250 mm with a particle size of 5 μm, containing *Lysimachia christinae* ODS C. 18 The column was used with a flow rate of 1 ml / min. High-performance preparative liquid chromatography (HPLC) was employed to separate and purify component F124H. The separated product was isocratically eluted with acetonitrile / water at a volume ratio of 40 / 60 at a retention time of 156 min to obtain the sesquiterpene dimer compound lemnalinoid L. The HPLC separation conditions were as follows: a 10 mm × 250 mm column with a particle size of 5 μm; and a *Lysimachia christinae* ODS C20 column. 18 The column was used at a flow rate of 2 ml / min.

[0030] Example 3 To determine the structure of the sesquiterpene dimer compound lemnalinoid L extracted by the preparation method provided in Example 2, this application subjected the sesquiterpene dimer compound lemnalinoid L extracted in Example 2 to ultraviolet spectroscopy, infrared spectroscopy, high-resolution mass spectrometry, and other methods. 1 H-NMR, 13 C-NMR, HSQC, HMBC, 1 H- 1 H COSY detection and ECD calculation determined its absolute configuration, and the attached... Figure 1 - Appendix Figure 11 And the one-dimensional and two-dimensional nuclear magnetic resonance data shown in Table 1.

[0031] From the appendix Figure 1 - Appendix Figure 3 It can be seen that in the sesquiterpene dimer compound lemnalinoid L, [ α ] 25 D +86.7 ( c 1.0,MeOH); UV (MeOH) λ max (log e ) = 193 (0.78) nm, λ max (log e = 220(1.20) nm; IR(KBr) n max =3415, 1716, 1595, 1363cm -1 HRESIMS shows that m / z 540.3328 of [M+H] + The quasi-molecular ion peak, with a calculated value of 540.3320, indicates that the molecular formula of this compound is C. 32 H 45 O6N has an unsaturation degree of 11.

[0032] Table 1: One-dimensional and two-dimensional NMR data of the sesquiterpene dimer compound lemnalinoid L (CDCl3, 500 MHz) From the appendix Figure 4 Appendix Figure 6 It can be seen that the sesquiterpene dimer lemnalinoid L has a single olefin proton signal in the low-field region. d H 5.51m, in the high field area d H 2.17, s; d H 1.20, s; d H 0.96, s, a total of 3 single-peak methyl signals and d H 1.98, d, J =1.4Hz; d H 1.54, d, J =7.4Hz; d H 1.06, d, J =6.8Hz; dH 0.99, d, J =7.1Hz, with a total of 4 bimodal methyl signals. (Observation of the attached...) Figure 5 Appendix Figure 7 As can be seen, the sesquiterpene dimer compound lemnalinoid L has 32 carbon signals, namely 7 methyl groups, 9 methylene groups, 6 methine groups, and 10 quaternary carbons. Among them, all 9 methylene groups are sp. 3 Hybridized carbon; 4 out of 6 methines are sp. 3 Hybridized carbon, 1 sp 2 Hybridized carbon and 1 oxygen-bound carbon; 2 sp atoms out of 10 quaternary carbons 3 Hybridized carbon, 3 sp 2 It has a hybrid carbon, one oxygen-bound carbon, three carbonyl groups, and one ketone group.

[0033] The following section analyzes the planar structure of the sesquiterpene dimer compound lemnalinoid L using NMR data. The details are as follows: According to the appendix Figure 8 The continuous proton correlation signals in the data can be used to connect three spin-coupled systems: H-1 / H-2 / H-3 / H-4 / H3-14, H-8 / H-9, and H-16 / H-18. According to the appendix... Figure 7 Relevant signals in: H3-15 ( d H 1.20) and C-4 ( d c34.3), C-5( d c45.3), C-6( d c159.1), C-10( d Related to c140.9); H2-9 ( d H 2.52, 2.16) and C-1 ( d c123.2), C-10( d Related to c140.9); H3-13 ( d H 1.98) and C-6 ( d c159.1), C-11( d c122.4), C-12( d Related to c172.9); H-7 ( d H 4.12) and C-6 ( d c159.1), C-12( d Related to c172.9); H-16 ( d H 4.58) and C-7 ( d c59.5), C-17 ( d (c171.1) is related, and can connect three spin-coupled systems, and construct such as by combining relevant chemical shift values. Figure 10 The planar structure of sesquiterpenes in part A is shown.

[0034] Subsequently, according to the appendix Figure 8 The continuous proton correlation signals in the data can connect two spin-coupled systems: H-1' / H-2' / H-3' / H-4' / H3-13' and H-8' / H-9'. According to the appendix... Figure 7 Relevant signals in: H3-14' ( d H 0.96) and C-4'( d c34.3), C-5'( d c45.7), C-6'( d c48.4), C-10'( d c89.6) related; H2-9'( d H 2.28, 2.16) and C-1'( d c73.4), C-10'( d c89.6) related; H2-8'( d H 2.64, 2.49) and C-7' ( d Related to c176.1), two spin-coupled systems can be connected, and a helical ring structure connecting six rings to five rings can be constructed by combining relevant chemical shift values. Then, according to the appendix... Figure 7 H3-12' (in the relevant signals) d H 2.17) and C-6' ( d c48.4), C-11'( d Related to c208.5), a methyl ketone fragment with a six-membered ring side chain can be constructed. Then, by combining chemical shift data, a fragment can be constructed as follows: Figure 10 Part B shows the sesquiterpene fragment. Finally, according to the appendix... Figure 7 H-1' (in the relevant signal) d H 4.98) and C-17 ( d Related to c171.1), the two sesquiterpene fragments A and B can be connected and combined, and the complete planar structure of the unsaturated compound lemnalinoid L can be constructed by combining the calculated structure.

[0035] The relative configuration of the sesquiterpene dimer compound lemnalinoid L is determined by the attachment Figure 9 To determine this. Specifically, for sesquiterpenes in part A, H3-15 ( d H1.20) and H3-14 d H 1.06), H-7 ( d H 4.12) Related, it can be determined that H-7, H3-14, and H3-15 are located on the same side of the ring. For part B sesquiterpenes, H-9'b ( d H 2.16) and H-1' ( d H 4.98), H3-13'( d H 0.99), H3-14'( d H Based on the correlation of 0.96), it can be determined that H-1', H-9'b, H3-13', and H3-14' are located on the same side of the ring. The relative configuration of the C-16 position in the remaining A portion cannot be determined by valid NOESY, but considering the NMR data alignment and the biosynthetic pathway, the C-16 configuration should be... S Therefore, the absolute configuration of the sesquiterpene dimer compound lemnalinoid L can be determined to be 4. S , 5 R , 7 S , 16 S , 1' S , 4' S , 5' R , 10' R Finally, through the appendix Figure 11 The comparison between the TDDFT-ECD calculation and the experimental CD value shown verifies the absolute configuration of the compound lemnalinoid L.

[0036] Example 4 The sesquiterpene dimer compound lemnalinoid L prepared in Example 2 of this application exhibits inhibitory activity against the human breast cancer cell line MDA-MB-231, and can serve as a candidate molecule for the preparation of antitumor drugs. To verify the inhibitory effect of the sesquiterpene dimer compound lemnalinoid L on the human breast cancer cell line MDA-MB-231, biological tests were conducted in this application, the details of which are as follows: The inhibitory effect of the sesquiterpene dimer compound lemnalinoid L prepared in Example 2 on human chronic myeloid leukemia cells K562 was tested by the MTT assay. The inhibitory effect of the sesquiterpene dimer compound lemnalinoid L on human normal hepatocytes L-02, human metastatic pancreatic adenocarcinoma cells ASPC-1, human breast cancer cells MDA-MB-231, human small cell lung cancer cells NCI-H446, and drug-resistant human small cell lung cancer cells NCI-H446 / EP was tested by the SRB assay. The positive control drug was doxorubicin.

[0037] In the initial screening test, the sesquiterpene dimer compound lemnalinoid L at a test concentration of 30 μM was preliminarily determined to have an inhibitory effect on human breast cancer cells MDA-MB-231. The test results are shown in Table 2. In the secondary screening test, the test concentrations of the sesquiterpene dimer compound lemnalinoid L were set at six levels: 30 μM, 15 μM, 7.5 μM, 3.25 μM, 1.875 μM, and 0.9375 μM. The secondary screening further verified that the sesquiterpene dimer compound lemnalinoid L had an inhibitory effect on human breast cancer cells MDA-MB-231 and determined its IC50. 50 Values; test results are shown in Table 3.

[0038] Table 2: Initial screening results of the inhibitory effect of the sesquiterpene dimer compound lemnalinoid L on tumor cells (inhibition rate %) Table 3: Screening results of the inhibitory effect of the sesquiterpene dimer compound lemnalinoid L on MDA-MB-231 cells As shown in Tables 2 and 3, the sesquiterpene dimer compound lemnalinoid L exhibits selective inhibitory activity against the human breast cancer cell line MDA-MB-231, with an IC50 value of [missing information]. 50 The value is 15.56 µ M. This indicates that the sesquiterpene dimer compound lemnalinoid L can be used to prepare drugs that inhibit human breast cancer cell lines.

[0039] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A sesquiterpene dimer compound, characterized in that, The chemical structural formula of the sesquiterpene dimer compound is as follows: 。 2. The method for preparing the sesquiterpene dimer compound according to claim 1, characterized in that, include: S01: Soft coral is cut into pieces and soaked in methanol at room temperature to obtain an extract. The extract is then concentrated under reduced pressure, desalted with anhydrous methanol, and concentrated under reduced pressure again to obtain a crude extract. S02: The crude extract was separated by silica gel vacuum column chromatography, gradient elution, thin-layer chromatography colorimetric detection, and the same fractions were combined and concentrated to obtain 16 primary components F1-F16; wherein, the gradient elution was performed sequentially with petroleum ether, petroleum ether / acetone in volume ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, dichloromethane, and dichloromethane / methanol in volume ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1; S03: F12 in the primary components F1-F16 was separated by silica gel column chromatography, gradient elution, thin-layer chromatography colorimetric detection, and the same fractions were combined and concentrated to obtain four secondary components F121-F124; wherein, gradient elution was performed sequentially using petroleum ether / acetone at volume ratios of 20 / 1, 10 / 1, 5 / 1, and 0 / 1. S04: After high performance liquid chromatography analysis, medium-pressure preparative liquid chromatography separation, and isocratic elution, tertiary components F124Q, F124F, and F124H were obtained from the secondary components F121-F124; wherein, isocratic elution was performed using methanol / water with a volume ratio of 80 / 20. S05: The tertiary component F124H was analyzed by high performance liquid chromatography, separated by high performance preparative liquid chromatography, and eluted isocratically to obtain a sesquiterpene dimer compound; wherein isocratically eluted with acetonitrile / water at a volume ratio of 40 / 60.

3. The method for preparing the sesquiterpene dimer compound according to claim 2, characterized in that, In S02, the silica gel reduced pressure column chromatography separation conditions are as follows: the chromatography column size is 160mm×500mm, and the silica gel particle size is 200-300 mesh; the thin layer chromatography colorimetric detection conditions are as follows: the developing solvent is petroleum ether and propanol in a volume ratio of 3:1, and the colorimetric reagent is a 10% sulfuric acid ethanol solution.

4. The method for preparing the sesquiterpene dimer compound according to claim 2, characterized in that, In S03, the silica gel column chromatography separation conditions are as follows: the column size is 60mm×500mm; the silica gel particle size is 300-400 mesh; the thin-layer chromatography colorimetric detection conditions are as follows: the developing solvent is petroleum ether and propanol in a volume ratio of 3:1, and the colorimetric reagent is a 10% sulfuric acid ethanol solution.

5. The method for preparing the sesquiterpene dimer compound according to claim 2, characterized in that, In S04, the high performance liquid chromatography analysis conditions are as follows: the analytical chromatographic column is Greenherb ODS C column with a size of 4.6 mm × 250 mm and a particle size of 5 μm, and the flow rate is 1 ml / min; the medium pressure preparative liquid chromatography separation conditions are as follows: the preparative chromatographic column is Greenherb ODS C column with a size of 20 mm × 250 mm and a particle size of 5 μm, and the flow rate is 10 ml / min. 18 column, and the flow rate is 1 ml / min; the medium pressure preparative liquid chromatography separation conditions are as follows: the preparative chromatographic column is Greenherb ODS C column with a size of 20 mm × 250 mm and a particle size of 5 μm, 18 column, and the flow rate is 10 ml / min.

6. The method for preparing the sesquiterpene dimer compound according to claim 2, characterized in that, In S05, the high performance liquid chromatography analysis conditions are as follows: the analytical chromatographic column is a Greenherb ODS C column with a size of 4.6 mm × 250 mm and a particle size of 5 μm, and the flow rate is 1 ml / min; the high performance preparative liquid chromatography separation conditions are as follows: the chromatographic column is a Greenherb ODS C column with a size of 10 mm × 250 mm and a particle size of 5 μm, and the flow rate is 2 ml / min. 18 column, and the flow rate is 1 ml / min; the high performance preparative liquid chromatography separation conditions are as follows: the chromatographic column is a Greenherb ODS C column with a size of 10 mm × 250 mm and a particle size of 5 μm, 18 column, and the flow rate is 2 ml / min.

7. The method for preparing the sesquiterpene dimer compound according to claim 2, characterized in that, In S04, the retention times of the tertiary components F124Q, F124F, and F124H in medium-pressure preparative liquid chromatography are 0.00-3.50 min, 3.50-7.60 min, and 7.60-9.50 min, respectively.

8. The method for preparing the sesquiterpene dimer compound according to claim 2, characterized in that, In S01, the temperature for the two-stage vacuum concentration process is 37°C and the pressure is 0.1 MPa.

9. The use of the sesquiterpene dimer compound of claim 1 or the sesquiterpene dimer compound prepared by any one of claims 2-8 in the preparation of a drug for inhibiting human breast cancer cell lines.

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