Sesquiterpenoids, preparation method thereof and application thereof in the field of anti-tumor

Abstract

The present invention discloses sesquiterpene compounds, a preparation method thereof and applications in the field of anti-tumor, belonging to the technical field of pharmaceutical compounds. The preparation method of the sesquiterpene compounds is as follows: (1) Take dry roots of Lindera glauca, slice them, add ethanol for extraction to obtain an extraction solution, combine the extraction solutions and concentrate to obtain an extract, and use petroleum ether to extract the extract to obtain a crude extract; (2) Purify the crude extract by normal-phase silica gel column chromatography, normal-phase silica gel column chromatography, reverse-phase silica gel column chromatography and semi-preparative HPLC to prepare the sesquiterpene compounds from the crude extract. The sesquiterpene compounds have certain anti-tumor activities and have good application prospects in the field of anti-tumor.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical compounds, and particularly relates to sesquiterpene compounds, a preparation method thereof, and their application in the field of anti-tumor. Background Art

[0002] Lindera glauca belongs to the genus Lindera of the Lauraceae family and is a shrub plant with the Latin name Lindera glauca (Siebold & Zucc.) Blume. Lindera glauca is mainly distributed in Shanxi, Shaanxi, Gansu, Henan, Sichuan, East China, Central South and other places in China. It has a long history of being used as medicine. Lindera glauca is pungent and very hot in taste. Its root is used to treat rheumatic arthralgia, overexertion, cold, tonsillitis, pharyngitis, edema, etc.

[0003] With the increase in the incidence and mortality of tumors, the development and isolation of anti-tumor drugs have become an urgent challenge. The complexity and heterogeneity of tumor cells make it difficult for traditional chemotherapy drugs to be effectively treated, and often drug resistance will occur. Natural small molecule compounds have unique advantages such as good curative effect and few side effects in anti-tumor, and are an important source for the discovery of new anti-tumor drugs.

[0004] For example, the sesquiterpene chamaejasmin A isolated by El-Desoky, A.H.H et al. from Stellera chamaejasme of the Thymelaeaceae family showed cytotoxic activity against cervical cancer HeLa cells, with an IC 50 value of 6.3 μmol·L -1 (El-Desoky, Ahmed H H et al. “Chamaejasmins, cytotoxic guaiane sesquiterpenes from the root of Stellera chamaejasme L.” Fitoterapia vol. 146 (2020): 104714.); Chen et al. isolated the compound 2β,7,3-trihydroxycalamenene 3-O-β-D-glucoside (110) from Abelmoschus sagittifolius and tested its effects on HeLa and HepG2 human cancer cell lines, and found that the compound had moderate cytotoxic activity, with IC 50 values of 12.88 μmol·L -1 and 18.15 μmol·L -1(Chen, De-Li et al. “A new cadinane sesquiterpenoid glucoside with cytotoxicity from Abelmoschus sagittifolius.” Natural product research vol. 33, 12 (2019): 1699-1704.); The Chinese patent document with the publication number CN110642822A discloses a group of sesquiterpenoid compounds, their preparation methods and applications. The sesquiterpenoid compounds in this invention are extracted from Wedelia trilobata and have anti-tumor and anti-inflammatory activities, and can be used to prepare anti-tumor drugs and anti-inflammatory drugs. The above studies all show that sesquiterpenoid compounds have great potential in the anti-tumor field.

[0005] The chemical constituents of the genus Lindera are mainly flavonoids, alkaloids and sesquiterpenoids, and sesquiterpenoid compounds are one of the characteristic chemical constituents of the genus Lindera. Therefore, the sesquiterpenoid compounds contained in Lindera glauca are worthy of in-depth exploration and research in order to provide more lead compounds for the research and development of new anti-tumor drugs. Summary of the Invention

[0006] The present invention provides a sesquiterpenoid compound, which is isolated from the tuberous roots of Lindera glauca, a plant of the genus Lindera in the Lauraceae family, and has certain anti-tumor activity.

[0007] The specific technical solutions adopted are as follows:

[0008] The sesquiterpenoid compound is selected from any one of the compounds (-)-1, (+)-1, compound 2, compound 3, compound 4, and compound 5 shown in the following formula:

[0009]

[0010] The sesquiterpenoid compound is isolated from the tuberous roots of Lindera glauca. The specific preparation method includes the following steps:

[0011] (1) Take the dried tuberous roots of Lindera glauca, slice them and extract with ethanol to obtain an extract. Combine the extracts and concentrate to obtain an extract paste. Extract the extract paste with petroleum ether to obtain a crude extract;

[0012] (2) Purify the crude extract by normal-phase silica gel column chromatography, reverse-phase silica gel column chromatography and semi-preparative HPLC to prepare compounds (-)-1, (+)-1, 2, 3, 4 and 5 from the crude extract.

[0013] Preferably, in step (1), the ethanol used is industrial ethanol with a concentration of 80-95%, the ethanol extraction method is the hot maceration method, the extraction temperature is 50-60°C, and the extraction is carried out 6-7 times, 20-24 hours each time.

[0014] More preferably, the ethanol used is industrial ethanol with a concentration of 95%, and the extraction is carried out at 55°C for 7 times, 24 hours each time.

[0015] Specifically, step (2) includes the following steps:

[0016] (2.1) The crude extract is separated by gradient elution on a normal-phase silica gel column, and the mobile phase is petroleum ether / ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is 40-0:1, and 7 fractions Fr.1-Fr.7 are collected;

[0017] (2.2) Fraction Fr.4 is separated by gradient elution on a normal-phase silica gel column, and the mobile phase is petroleum ether / ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is 12-0:1. The eluate is detected by thin-layer chromatography, and the same components are combined to obtain 21 fractions Fr.4.1-Fr.4.21;

[0018] (2.3) Fraction Fr.4.5 is separated by gradient elution on a reverse-phase silica gel column, and the mobile phase is methanol / water, and the volume ratio of methanol to water is 60-100:40-0, and 54 fractions Fr.4.5.1-Fr.4.5.54 are collected; Fraction Fr.4.5.4 is purified by semi-preparative HPLC to obtain compound 4 and compound 5;

[0019] (2.4) Fraction Fr.4.12 is separated by gradient elution on a reverse-phase silica gel column, and the mobile phase is methanol / water, and the volume ratio of methanol to water is 20-100:80-0, and 45 fractions Fr.4.12.1-Fr.4.12.45 are collected;

[0020] (2.5) Fraction Fr.4.12.8 is purified by semi-preparative HPLC three times to obtain compound 2 and compound 3; Fraction Fr.4.12.9 is purified by semi-preparative HPLC twice to obtain compound (-)-1 and compound (+)-1.

[0021] Preferably, in step (2.1), the volume ratio change of the petroleum ether / ethyl acetate gradient elution is 40:1, 20:1, 10:1, 5:1, 2:1, 1:1, 0:1;

[0022] In step (2.2), the volume ratio change of the petroleum ether / ethyl acetate gradient elution is 12:1, 10:1, 8:1, 5:1, 3:1, 0:1;

[0023] In step (2.3), the volume ratio change of methanol / water gradient elution is 60:40, 70:30, 80:20, 90:10, 100:0;

[0024] In step (2.4), the volume ratio change of methanol / water gradient elution is 20:80, 40:60, 60:40, 80:20, 100:0.

[0025] Preferably, in step (2.1), the silica gel column chromatography is normal-phase silica gel column chromatography, using 80 - 120 mesh silica gel particles.

[0026] Preferably, in step (2.2), the normal-phase silica gel column uses 200 - 300 mesh silica gel particles as the packing material.

[0027] Preferably, in steps (2.3) and (2.4), the reverse-phase silica gel column uses YMC GEL ODS-A-HG C18 silica gel packing material.

[0028] Preferably, in step (2.3), the conditions of semi-preparative HPLC method are: Welch Ultimate XB-C18 liquid chromatography column, isocratic elution separation, the mobile phase is acetonitrile / water with a volume ratio of 65:35, and the flow rate is 2.0 mL / min.

[0029] Preferably, in step (2.5), the conditions for purifying compound 2 and compound 3 by three times of semi-preparative HPLC method are: for the first semi-preparative HPLC method, Welch Ultimate XB-C18 liquid chromatography column is selected, isocratic elution separation, the mobile phase is acetonitrile / water with a volume ratio of 35:65, and the flow rate is 2.0 mL / min; for the second semi-preparative HPLC method, Welch Ultimate XB-C18 liquid chromatography column is selected, isocratic elution separation, the mobile phase is methanol / water with a volume ratio of 40:60, and the flow rate is 2.0 mL / min; for the third semi-preparative HPLC method, Daicel IC column is selected, isocratic elution separation, the mobile phase is methanol / water with a volume ratio of 60:40, and the flow rate is 1.0 mL / min.

[0030] Preferably, in step (2.5), the conditions for purifying compound (-)-1 and compound (+)-1 by two times of semi-preparative HPLC method are: for the first semi-preparative HPLC method, Welch Ultimate XB-C18 liquid chromatography column is used, isocratic elution separation, the mobile phase is acetonitrile / water with a volume ratio of 36:64, and the flow rate is 2.0 mL / min; for the second semi-preparative HPLC method, Daicel IC column is selected, isocratic elution separation, the mobile phase is acetonitrile / water with a volume ratio of 25:75, and the flow rate is 1.0 mL / min.

[0031] The present invention also discloses the application of the sesquiterpene compounds in the field of anti-tumor.

[0032] The present invention also discloses an anti-tumor drug, which comprises the sesquiterpene compounds, and has an inhibitory effect on Bcap-37 (human breast cancer cells) and DU-145 (human prostate cancer cells).

[0033] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0034] (1) The sesquiterpene compounds provided in the present invention have not been reported in the prior art. They are all optically pure compounds with determined stereoconfigurations, and have strong anti-tumor activities, showing good application prospects in the field of anti-tumor.

[0035] (2) The preparation method of the present invention is simple and easy to implement. It only needs to use normal-phase silica gel column, reverse-phase silica gel column and chromatographic column for separation, with less reagent consumption, good reproducibility and high purity. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 is the HRESIMS spectrum of Compound 1;

[0037] Figure 2 is the HSQC spectrum of Compound 1;

[0038] Figure 3 is of Compound 1 1 H- 1 H COSY spectrum;

[0039] Figure 4 is the HMBC spectrum of Compound 1;

[0040] Figure 5 is the NOESY spectrum of Compound 1;

[0041] Figure 6 is the experimental and calculated ECD diagrams of Compound (-)-1 and Compound (+)-1;

[0042] Figure 7 is the HRESIMS spectrum of Compound 5;

[0043] Figure 8 is the HSQC spectrum of Compound 5;

[0044] Figure 9 is of Compound 5 1 H- 1 H COSY spectrum;

[0045] Figure 10 is the HMBC spectrum of Compound 5;

[0046] Figure 11 1H-1H NOESY spectrum of Compound 5

[0047] Figure 12 X-ray single crystal diffraction pattern of Compound 5 Detailed implementation manners

[0048] The present invention will be further illustrated below in conjunction with embodiments and drawings. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention.

[0049] The roots of Lindera glauca used in the examples are from Lindera glauca (Siebold & Zucc.) Blume of the genus Lindera in the Lauraceae family.

[0050] Example 1

[0051] Take 45 kg of dried roots of Lindera glauca, slice them, and perform hot maceration extraction with 75 L of ethanol (80%-95% industrial ethanol) each time. The extraction temperature is 55 °C, and the extraction is carried out 7 times, 24 hours each time; combine the extraction solutions and concentrate to obtain an extract, and extract the extract with petroleum ether to obtain a crude extract;

[0052] Perform gradient elution separation of the crude extract by normal-phase silica gel column chromatography (silica gel particles are 80-120 mesh). The mobile phase is petroleum ether / ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is 40:1, 20:1, 10:1, 5:1, 2:1, 1:1, 0:1. Seven fractions Fr.1-Fr.7 are collected;

[0053] Perform gradient elution separation of fraction Fr.4 by normal-phase silica gel column chromatography. The mobile phase is petroleum ether / ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate is 12:1, 10:1, 8:1, 5:1, 3:1, 0:1. The eluate is detected by thin-layer chromatography, and the same components are combined to obtain 21 fractions Fr.4.1-Fr.4.21;

[0054] Perform gradient elution separation of fraction Fr.4.5 by reverse-phase silica gel column chromatography (YMC GEL ODS-A-HG C18 silica gel packing). The mobile phase is methanol / water, and the volume ratio of methanol to water is 60:40, 70:30, 80:20, 90:10, 100:0. Fifty-four fractions Fr.4.5.1-Fr.4.5.54 are collected;

[0055] Fraction Fr.4.5.4 is purified by semi-preparative HPLC method (Welch Ultimate XB-C18 liquid chromatography column, MeCN / H2O, 65:35, v / v, 2.0 mL / min) to obtain Compound 4 and Compound 5;

[0056] Fraction Fr.4.12 was separated by gradient elution through reverse-phase silica gel column chromatography (YMC GEL ODS-A-HG C18 silica gel packing material), with the mobile phase being methanol / water, and the volume ratio of methanol to water being 20:80, 40:60, 60:40, 80:20, 100:0, and 45 fractions Fr.4.12.1 - Fr.4.12.45 were collected;

[0057] Fraction Fr.4.12.8 was further separated on semi-preparative HPLC (Welch Ultimate XB-C18 liquid chromatography column, MeCN / H2O, 35:65, v / v, 2.0 mL / min) to obtain liquid chromatography peaks Fr.4.12.8.1 - Fr.4.12.8.18; Liquid chromatography peak Fr.4.12.8.8 was separated on semi-preparative HPLC (Welch Ultimate XB-C18 liquid chromatography column, MeOH / H2O, 40:60, v / v, 2.0 mL / min) to obtain liquid chromatography peak Fr.4.12.8.8.3; Liquid chromatography peak Fr.4.12.8.8.3 was purified again by semi-preparative HPLC (Daicel IC column, MeOH / H2O, 60:40, v / v, 1.0 mL / min) to obtain Compound 3 and Compound 2;

[0058] Fraction Fr.4.12.9 was further separated on semi-preparative HPLC (Welch Ultimate XB-C18 liquid chromatography column, MeCN / H2O, 36:64, v / v, 2.0 mL / min) to obtain liquid chromatography peaks Fr.4.12.9.1 - Fr.4.12.9.12, and liquid chromatography peak Fr.4.12.9.10 was purified on semi-preparative HPLC (Daicel IC column, MeCN / H2O, 25:75, v / v, 1.0 mL / min) to obtain Compound (-)-1 and (+)-1.

[0059] Sample analysis

[0060] (1) Structure identification

[0061] The structures of Compounds 1 (including (-)-1 and (+)-1), 2, 3, 4, and 5 were identified using a variety of spectroscopic techniques, electronic circular dichroism (ECD) theoretical calculations, and single crystal derivatives.

[0062] Compound 1: White amorphous powder; UV (MeOH) λ max (logε) = 207 (4.16) nm; IR (KBr) ν max = 3448, 2925, 2360, 1767, 1715, 1636, 1384, 1148 cm-1 ; ECD (MeOH) λ max (Δε) = 201 (-12.36), 270 (+16.44) nm; As Figure 1 shown, HRESIMS gave the quasi-molecular ion peak [M+Na] + m / z 243.1365 (calcd for C 14 H 20 O2Na + , 243.1361), molecular weight was 220.1463, and the molecular formula was C 14 H 20 O2.

[0063] For compound 1 1 H (600 MHz) and 13 C (150 MHz) NMR data (CDCl3) indicated that compounds (-)-1 and (+)-1 were a pair of enantiomers, and their 1 H NMR and 13 C NMR results were the same as those of compound 1;

[0064] The HSQC spectrum, 1 H- 1 H COSY spectrum, HMBC spectrum, and NOESY spectrum of compound 1 were further measured ( Figures 2 - 5 ), and the planar structure of 1 was determined. The optical rotation of compound 1 was nearly zero to guide chiral separation, and the results showed that compound 1 was a pair of enantiomers.

[0065] To determine the absolute configuration of (±)-1, the calculated ECDs of 4 possible epimers (S,S-1, R,S-1, S,R-1, R,R-1) were measured. As Figure 6 shown, based on the good consistency trend between the experimental and calculated ECD curves, (-)-1 and (+)-1 were finally determined to be 7R,10R and 7S,10S.

[0066] It was tested that (-)-1: ECD (MeOH) λ max (Δε) = 294 (-25.44) nm; (+)-1: ECD (MeOH) λ max (Δε) = 296 (-23.82) nm.

[0067] Compound 2: White amorphous powder; UV (MeOH) λ max (logε) = 239 (4.43) nm; IR (KBr) ν max= 3422, 2921, 2851, 1702, 1647, 1395, 1288, 1231, 1171 cm -1 ; ECD (MeOH) λ max (Δε) = 192 (-14.39), 206 (+7.63), 228 (-10.02), 249 (+2.03), 278 (-12.40) nm; HRESIMS [M+H] + m / z 221.1534 (calcd for C 14 H 21 O2 + , 221.1542), molecular weight 220.1463, molecular formula C 14 H 20 O2.

[0068] Compound 3: White amorphous powder; UV (MeOH) λmax (logε) = 241 (4.06) nm; IR (KBr) ν max = 3356, 2920, 2850, 2360, 2342, 1697, 1636, 1470, 1457 cm -1 ; ECD (MeOH) λ max (Δε) = 211 (-15.80), 256 (+2.06), 295 (-6.29) nm; HRESIMS [M+H] + m / z 221.1542 (calcd for C 14 H 21 O2 + , 221.1542), molecular weight 220.1463, molecular formula C 14 H 20 O2.

[0069] For compounds 2 and 3 1 H (600 MHz) and 13 C (150 MHz) NMR data (CDCl3) are shown in Table 1:

[0070] Table 1 Test data of 1 H NMR and 13 C NMR for compounds 2 and 3

[0071]

[0072] The HSQC spectra, 1 H- 1 H COSY spectra, HMBC spectra and NOESY spectra of compounds 2 and 3 were further tested; By 13The relative and absolute configurations of compounds 2 and 3 were determined by CNMR and ECD calculations. The absolute configurations of 2 and 3 are 4S,7R,10S and 4R,7S,10S, respectively.

[0073] Compound 4: Colorless cubic crystals; mp 178 - 179 °C; UV(MeOH)λ max (logε) = 198(4.68) nm; IR(KBr)ν max = 2926, 2360, 1677, 1621, 1436, 1277, 1237 cm -1 ; ECD(MeOH)λ max (Δε) = 197(-15.88), and 241(-5.21) nm; HRESIMS[M+Na] + m / z 273.1468 (calcd for C 15 H 22 O3Na + , 273.1467), molecular weight is 250.1569, and the molecular formula is C 15 H 22 O3.

[0074] 1H NMR (600 MHz) and 1 13C NMR (150 MHz) data of compound 4 (CDCl3) are shown in Table 2: 13

[0075] Table 2 1H NMR and 1 13C NMR test data of compound 4 13 Based on one - dimensional and two - dimensional NMR data, it was confirmed that compound 4 is a guaiane - type sesquiterpenoid compound. The single - crystal X - ray diffraction pattern (CCDC 2302537) was obtained using a Cu target single - crystal diffraction technique to analyze the absolute configuration of compound 4, and the absolute configuration of 4 was determined to be 1R,4S,5R,7R,10S.

[0076]

[0077] Compound 5: Colorless needle - shaped crystals; mp 178 - 179 °C;

[0078] UV(MeOH)λ UV(MeOH)λ max (logε) = 200(4.06) nm; IR(KBr)ν max = 3210, 2973, 2926, 2361, 1720, 1676, 1622, 1412, 1385, 1202, 1173 cm -1 ; ECD(MeOH)λmax (Δε) = 199 (-15.83), 224 (-1.89) and 231 (-7.84) nm; as Figure 7 shown, HRESIMS [M-H] - m / z 293.1763 (calcd for C 17 H 25 O4 - , 293.1753), the molecular weight is 294.1831, and the molecular formula is C 17 H 24 O4.

[0079] For compound 5 1 H (600 MHz) and 13 C (150 MHz) NMR data (CDCl3) are shown in Table 3:

[0080] Table 3 1 1H NMR and 13 13C NMR test data of compound 5

[0081]

[0082] The NMR data of compound 5 are similar to those of compound 4, indicating that the structure of compound 5 is similar to that of compound 4. The HSQC spectrum, 1 1H- 1 1H COSY spectrum, HMBC spectrum and NOESY spectrum of compound 5 were further measured ( Figures 8 - 11 ); A single crystal X-ray diffraction experiment (CCDC 2302538) was carried out, and the results are as Figure 12 shown, confirming that the absolute configuration of compound 5 is 1R, 4S, 5R, 7R, 10S.

[0083] (2) Antitumor activity

[0084] The cytotoxicity of compounds (-)-1, (+)-1, 2, 3, 4, and 5 against Du-145, A549, HepG2, HCT116, and three breast cancer cell lines (Bcap-37, MCF-7, and MDA-MB-231) was determined using CCK-8. Breast cancer cells (Bcap-37, MCF-7, and MDA-MB-231), Du-145, A549, HepG2, and HCT116 cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM, Gibco) containing 10% fetal bovine serum (FBS, Gibco), 100 U / mL penicillin, and 100 μg / mL streptomycin (Beyotime biotech). And they were cultured in a 5% CO2 incubator at 37 °C. After culturing, breast cancer cells (Bcap-37, MCF-7, and MDA-MB-231), Du-145, A549, HepG2, and HCT116 (1×10 4 cells) were seeded into 96-well plates and treated with different concentrations (2.5, 5, 10, 20, 40 μM) of the compounds for 24 hours. Cell Counting Kit-8 (CCK-8, Biosharp Life sciences) was used to detect cytotoxicity according to the instructions.

[0085] As shown in Table 4, the results showed that compounds (-)-1 and 5 selectively inhibited Bcap-37 and Du-145 cells, with IC 50 values of 5.60 ± 0.228 and 5.52 ± 1.96 μM, respectively, and the results showed that they were significantly superior to the positive controls (cisplatin and paclitaxel). The other compounds were inactive against these cell lines at 40 μM.

[0086] Table 4 Results of cytotoxicity experiments of sesquiterpene compounds against tumor cells

[0087]

[0088] The above-described embodiments have described the technical solutions of the present invention in detail. It should be understood that the above are only specific embodiments of the present invention and are not used to limit the present invention. Any modifications, supplements, or substitutions in a similar manner within the scope of the principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Sesquiterpene compounds, characterized in that, Either compound (-)-1 or compound 5 selected from the compounds shown by the following formula:

2. The preparation method of the sesquiterpene compound according to claim 1, wherein Comprising the following steps: (1) Take dry Lindera aggregata roots, slice them, extract with ethanol to obtain an extract, combine the extracts and concentrate to obtain an extract paste, and extract the extract paste with petroleum ether to obtain a crude extract; (2) Purify the crude extract by normal-phase silica gel column chromatography, reverse-phase silica gel column chromatography and semi-preparative HPLC to prepare compounds (-)-1 and 5 from the crude extract; Step (2) includes the following steps: (2.1) Gradient elution separation of the crude extract by normal-phase silica gel column chromatography, with the mobile phase being petroleum ether / ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate being 40-0:1, and collect 7 fractions Fr.1-Fr.7; (2.2) Gradient elution separation of fraction Fr.4 by normal-phase silica gel column chromatography, with the mobile phase being petroleum ether / ethyl acetate, and the volume ratio of petroleum ether to ethyl acetate being 12-0:1, and detect the eluate by thin-layer chromatography, combine the same components, and obtain 21 fractions Fr.4.1-Fr.4.21; (2.3) Gradient elution separation of fraction Fr.4.5 by reverse-phase silica gel column chromatography, with the mobile phase being methanol / water, and the volume ratio of methanol to water being 60-100:40-0, and collect 54 fractions Fr.4.5.1-Fr.4.5.54; fraction Fr.4.5.4 is purified by semi-preparative HPLC to obtain compound 5; (2.4) Gradient elution separation of fraction Fr.4.12 by reverse-phase silica gel column chromatography, with the mobile phase being methanol / water, and the volume ratio of methanol to water being 20-100:80-0, and collect 45 fractions Fr.4.12.1-Fr.4.12.45; (2.5) Fraction Fr.4.12.9 is purified by semi-preparative HPLC twice to obtain compound (-)-1.

3. The preparation method of the sesquiterpene compound according to claim 2, wherein In step (1), during the ethanol extraction process, the extraction temperature is 50-60 °C, the extraction is carried out 6-7 times, and each time is 20-24 hours.

4. The method for preparing sesquiterpenoid compounds according to claim 2, wherein In step (2.1), the volume ratio change of the petroleum ether / ethyl acetate gradient elution is 40:1, 20:1, 10:1, 5:1, 2:1, 1:1, 0:1; In step (2.2), the volume ratio change of the petroleum ether / ethyl acetate gradient elution is 12:1, 10:1, 8:1, 5:1, 3:1, 0:1; In step (2.3), the volume ratio change of the methanol / water gradient elution is 60:40, 70:30, 80:20, 90:10, 100:0; In step (2.4), the volume ratio change of the methanol / water gradient elution is 20:80, 40:60, 60:40, 80:20, 100:

0.

5. The method for preparing sesquiterpenoid compounds according to claim 2, wherein In step (2.3), the conditions of the semi-preparative HPLC method are: Welch Ultimate XB-C18 liquid chromatography column, isocratic elution separation, and the mobile phase is acetonitrile / water with a volume ratio of 65:

35.

6. The preparation method of the sesquiterpene compound according to claim 2, wherein In step (2.5), the conditions for purifying the compound (-)-1 by semi-preparative HPLC twice are as follows: for the first semi-preparative HPLC, a Welch Ultimate XB-C18 liquid chromatography column is selected, and isocratic elution separation is carried out, and the mobile phase is acetonitrile / water with a volume ratio of 36:64; for the second semi-preparative HPLC, a Daicel IC column is selected, and isocratic elution separation is carried out, and the mobile phase is acetonitrile / water with a volume ratio of 25:

75.

7. Use of the sesquiterpenoid compound according to claim 1 in the preparation of an anti-tumor drug, characterized in that, The compound (-)-1 is used for preparing an anti-breast cancer drug, and the compound 5 is used for preparing an anti-prostate cancer drug.

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