New enantiokaurane compound with anti-tumor activity as well as preparation method and application of new enantiokaurane compound

Abstract

The invention discloses an enantiokaurane compound with antitumor activity as well as a preparation method and application of the enantiokaurane compound. According to the present invention, chemical components of Western African plant I.trichantha tubers are deeply studied, and a variety of chromatographic separation methods such as macroporous resin, silica gel resin, medium pressure preparation-MCI column, gel normal pressure column and semi-preparative high performance liquid chromatography are combined to separate to obtain one enantiokaurane new compound Trichanthone B; experiments show that the compound obtained through separation has a good inhibition effect on an MIA PaCa-2 pancreatic cancer cell line and an A549 lung cancer cell line, and IC50 of the compound on the MIA PaCa-2 pancreatic cancer cell line reaches a nanomole level; the effect of the compound is stronger than that of positive drugs 5-fluorouracil and carboplatin, and the compound has the potential of being developed into new drugs for resisting pancreatic cancer and lung cancer.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a new enantiokaurane compound, a preparation method thereof, and a new anti-tumor use thereof. Background Art

[0002] I. trichantha is a plant of the genus Cornus that is endemic to Central and West Africa and is a medicinal plant used in Nigeria and neighboring countries. It plays an important role in traditional Nigerian medicine, with local people using its tubers for a wide range of ailments, including poisoning, constipation, vomiting, and malaria. The tubers of I. trichantha are rich in starch, and during years of famine, local people would harvest and repeatedly wash the tubers before eating them. Modern research indicates that I. trichantha possesses a variety of physiological activities, including hypoglycemic, anticonvulsant, sedative, analgesic, and antibacterial properties. Currently, little research has been conducted on the chemical composition of I. trichantha.

[0003] Therefore, it is necessary to conduct in-depth research on the active ingredients of I. trichantha tubers based on existing technologies to develop new active ingredients and provide a scientific basis for expanding their clinical use. Summary of the Invention

[0004] The purpose of the present invention is to conduct in-depth research on the active ingredients of I. trichantha tubers, extract and separate the active ingredients with potential activity, and provide a scientific basis for clinical application.

[0005] Technical solution: The present invention isolates a new enantiokaurane compound from the tuber of the plant I. trichantha, and the compound is named Trichanthone B.

[0006] The structural formula of the compound of the present invention is:

[0007]

[0008] The method for preparing a new enantio-kaurane compound of the present invention is characterized by comprising the following steps:

[0009] (1) Weigh the dried I. trichantha tubers, crush them, add ethanol and water to reflux and extract the medicinal materials, filter and collect the filtrate, and concentrate under reduced pressure until there is no alcohol smell to obtain a concentrate;

[0010] (2) extracting the concentrated solution obtained in step (1) with petroleum ether, dichloromethane, ethyl acetate and n-butanol, respectively, and concentrating under reduced pressure to obtain a petroleum ether fraction, a dichloromethane fraction, an ethyl acetate fraction, an n-butanol fraction and a raffinate fraction, respectively;

[0011] (3) The dichloromethane fraction obtained in step (2) was subjected to atmospheric pressure column chromatography using AB-8 macroporous resin and eluted with ethanol-water to obtain three sub-fractions DCM-1 to DCM-3;

[0012] (4) The eluate fraction DCM-3 obtained in step (3) was subjected to silica gel atmospheric pressure column chromatography and eluted with dichloromethane-methanol to obtain 7 sub-fractions DCM-3-1-DCM-3-7;

[0013] (5) The fraction DCM-3-2 obtained in step (4) was subjected to medium pressure preparative-MCI column chromatography with gradient elution using methanol-water as the mobile phase to obtain 7 fractions DCM-3-2-1-DCM-3-2-7;

[0014] (6) The fraction DCM-3-2-1 obtained in step (5) was subjected to gel column chromatography with gradient elution using methanol-water as the mobile phase to obtain two sub-fractions DCM-3-2-1-1-DCM-3-2-1-2; the fraction DCM-3-2-1-1 was subjected to semi-preparative high performance liquid chromatography to obtain the compound.

[0015] As a more limited solution, the preparation method of the present invention comprises the following steps:

[0016] (1) Weigh the dried tubers of I. trichantha, crush them, and add 95% ethanol by volume to reflux extract the medicinal material. The solid-liquid ratio is 1:6-10. Extract 1-3 times, each time for 1-3 hours. Filter and collect the filtrate. Combine the filtrates and concentrate under reduced pressure until there is no alcohol taste to obtain an extract.

[0017] (2) The extract obtained in step (1) is made into a suspension with an appropriate amount of water, and then extracted with equal volumes of petroleum ether, dichloromethane, ethyl acetate and n-butanol, each extracted 3 to 6 times, to obtain a petroleum ether fraction, a dichloromethane fraction, an ethyl acetate fraction, an n-butanol fraction and a raffinate water fraction, respectively.

[0018] (3) The dichloromethane fraction obtained in step (2) was mixed and subjected to macroporous resin atmospheric pressure column chromatography, with gradient elution using ethanol-water mobile phases in volume ratios of 30:70, 60:40, 90:10, and 100:0, with each gradient elution lasting 5 column volumes; after thin layer chromatography and ultra-high performance liquid chromatography analysis, the fractions were concentrated and combined using a rotary evaporator to obtain three sub-fractions DCM-1 to DCM-3;

[0019] (4) The eluate fraction DCM-3 obtained in step (3) was mixed with a sample and subjected to silica gel atmospheric pressure column chromatography, with dichloromethane-methanol as the mobile phase in a volume ratio of 100:0, 50:1, 30:1, 20:1, 10:1, 7:1, 5:1, 3:1, 2:1, 1:1, 1:3, 1:7, and 0:100 as the gradient elution, with each gradient elution lasting 4 column volumes; after thin layer chromatography and ultra-performance liquid chromatography analysis, the fractions were concentrated and combined using a rotary evaporator to obtain 7 sub-fractions DCM-3-1-DCM-3-7;

[0020] (5) The fraction DCM-3-2 obtained in step (4) was mixed and subjected to gradient elution on a medium pressure preparative MCI column chromatography with methanol-water as the mobile phase, wherein A was water and B was methanol; the elution gradient program was: 0.01-20.00 min, 25%-25% B; 20.00-50.00 min, 35%-35% B; 50.00-80.00 min, 37%-37% B; 80.00-110.00 min, 38%-38% B; 110.00-140.00 min, 39%-39% B; 140.00-170.00 min, 41%-41% B; 170.00-200.00 min, 44%-44% B; 200.00-230.00 min, 49%-49% B; 230.00-260.00 min, 60%-60% B; 260.00-300.00 min, 100%-100% B, elution flow rate 23 mL / min, detection wavelengths 240 nm and 310 nm; after ultra-performance liquid chromatography analysis, the concentrated and combined fractions obtained were 7 fractions DCM-3-2-1-DCM-3-2-7;

[0021] (6) The 150-170 min eluate fraction DCM-3-2-1 obtained in step (5) was subjected to gel column chromatography at atmospheric pressure, eluted with methanol, analyzed by thin layer chromatography and ultra-high performance liquid chromatography, and then concentrated and combined using a rotary evaporator to obtain two sub-fractions DCM-3-2-1 and DCM-3-2-2;

[0022] (7) The eluate fraction DCM-3-2-1-1 obtained in step (6) was subjected to semi-preparative high performance liquid chromatography to obtain a monomer compound.

[0023] As a preferred embodiment, the chromatographic conditions for the semi-preparative HPLC described above are: an X-Bridge C18 preparative column (10 nm, 5 μm, 10×250 mm), an HPLC column pressure of 9.8 MPa, a column temperature of 22-26°C, an injection volume of 60 μL, a mobile phase of MeOH-H2O (v / v, 30:70), a flow rate of 3 mL / min, and detection wavelengths of 240 nm and 310 nm. The monomer compound Trichanthone B was obtained.

[0024] Beneficial effects:

[0025] This study, conducted through in-depth research on the chemical components of the tubers of the West African plant I. trichantha, isolated a new enantiokaurane compound, Trichanthone B, by combining multiple chromatographic separation methods, including macroporous resin, silica gel resin, medium-pressure preparative MCI column, gel atmospheric pressure column, and semi-preparative high-performance liquid chromatography. Pharmaceutical experiments revealed that the compound prepared by the present invention exhibits significant anti-tumor activity (including against pancreatic cancer, colon cancer, lung cancer, or liver cancer).

[0026] The compound Trichanthone B obtained by extraction and separation of the present invention can be prepared into various dosage forms with pharmaceutically acceptable carriers. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 (+)-HR-ESI-MS spectrum of the compound;

[0028] Figure 2 UV images of compounds;

[0029] Figure 3 Compound 1 H NMR spectrum (500 MHz, Methanol-d4);

[0030] Figure 4 Compound 13 C NMR spectrum (125 MHz, Methanol-d4);

[0031] Figure 5 DEPT 135 pattern of the compound (125 MHz, Methanol-d4);

[0032] Figure 6 Compound 1 H- 1 H COSY diagram (500MHz, Methanol-d4);

[0033] Figure 7 HSQC diagram of the compound ( 1 H:500MHz,13 C:125MHz,Methanol-d4);

[0034] Figure 8 The HMBC diagram of the compound ( 1 H:500MHz, 13 C:125MHz,Methanol-d4);

[0035] Figure 9 NOSEY pattern of the compound (500 MHz, Methanol-d4);

[0036] Figure 10 Compound 1 H- 1 Correlation plots of H COSY, key HMBC, and NOESY. DETAILED DESCRIPTION

[0037] Example 1

[0038] 1. Instruments and Materials

[0039] 1.1 Instrument

[0040]

[0041] Experimental Materials

[0042] X-Bridge C18 preparative column (10 nm, 5 μm, 10 × 250 mm); Waters ACQUITY UPLC BEH C 18 (2.1 mm × 100 mm, 1.7 μm) chromatographic column; MCI GEL (CHP20, 75–150 μm), column chromatography silica gel (200–300 mesh); column chromatography gel (Sephadex LH-20); chromatographic grade acetonitrile, methanol, and formic acid were purchased from Merck, USA; analytical grade petroleum ether, ethyl acetate, etc. were purchased from Nanjing Wanqing Chemical Reagent Co., Ltd.

[0043] 2. The preparation method of the compound is carried out according to the following steps

[0044] (1) Weigh the dried I. trichantha tubers, crush them, and add 95% ethanol (volume concentration) to reflux extract the medicinal material. The solid-liquid ratio is 1:10. Extract three times, each time for 2.5 h. Filter and collect the filtrate. Combine the filtrates and concentrate under reduced pressure until there is no alcohol smell to obtain an extract.

[0045] (2) The extract obtained in step (1) is made into a suspension with an appropriate amount of water, and then extracted with equal volumes of petroleum ether, dichloromethane, ethyl acetate and n-butanol, each extracted 5 times, to obtain a petroleum ether fraction, a dichloromethane fraction, an ethyl acetate fraction, an n-butanol fraction and a raffinate fraction, respectively.

[0046] (3) The dichloromethane fraction obtained in step (2) was mixed with the sample and macroporous resin at a ratio of 1:1.5, and subjected to macroporous resin atmospheric pressure column chromatography, with ethanol-water mobile phases having a volume ratio of 30:70, 60:40, 90:10, and 100:0, with each gradient elution lasting 5 column volumes. After thin layer chromatography and ultra-high performance liquid chromatography analysis, the fractions were concentrated and combined using a rotary evaporator to obtain three sub-fractions, DCM-1 to DCM-3.

[0047] (4) The eluate fraction DCM-3 obtained in step (3) was mixed with sample: silica gel at a ratio of 1:1.5, and subjected to silica gel atmospheric pressure column chromatography, with dichloromethane-methanol as the mobile phase in a volume ratio of 100:0, 50:1, 30:1, 20:1, 10:1, 7:1, 5:1, 3:1, 2:1, 1:1, 1:3, 1:7, and 0:100 as the gradient elution, with each gradient elution lasting 4 column volumes; after thin layer chromatography and ultra-performance liquid chromatography analysis, the fractions were concentrated and combined using a rotary evaporator to obtain 7 sub-fractions DCM-3-1-DCM-3-7;

[0048] (5) The fraction DCM-3-2 obtained in step (4) was mixed with the sample:MCI=1:1.2, and gradient elution was performed on a medium pressure preparative-MCI column chromatography with methanol-water as the mobile phase, wherein A is water and B is methanol; the elution gradient program was: 0.01-20.00 min, 25%-25% B; 20.00-50.00 min, 35%-35% B; 50.00-80.00 min, 37%-37% B; 80.00-110.00 min, 38%-38% B; 110.00-140.00 min, 39%-39% B; 140.00-170.00 min, 41%-41% B; 170.00-200.00 min, 44%-44% B; 200.00-230.00 min, 49%-49% B; 230.00-260.00 min, 60%-60% B; 260.00-300.00 min, 100%-100% B, elution rate 23 mL / min, detection wavelengths 240 nm and 310 nm; after ultra-performance liquid chromatography analysis, the products were concentrated and combined to obtain 7 fractions DCM-3-2-1-DCM-3-2-7;

[0049] (6) The 150-170 min eluate fraction DCM-3-2-1 obtained in step (5) was subjected to gel column chromatography at atmospheric pressure, eluted with methanol, analyzed by thin layer chromatography and ultra-high performance liquid chromatography, and then concentrated and combined using a rotary evaporator to obtain two sub-fractions DCM-3-2-1-1-DCM-3-2-1-2;

[0050] (7) The eluate fraction DCM-3-2-1-1 obtained in step (6) was subjected to semi-preparative high performance liquid chromatography using pure water (A)-methanol (B) as the mobile phase (MeOH-H2O, v / v, 30:70) to obtain the monomer compound in the 38-42 min period.

[0051] 3. Structural analysis of compounds

[0052] 3.1 Structural identification of Trichanthone B(1)

[0053]

[0054] The compound is a white amorphous powder. (c 0.01, MeOH). Combining 13C NMR data with high resolution mass spectrometry (HRESIMS) gave a quasi-molecular ion peak m / z 309.1701 ([M+H] + , C 17 H 25 O5 + The calculated value is 309.1704), and the molecular formula of the compound can be deduced to be C 17 H 24 O5, the degree of unsaturation is 6. The hydrogen spectrum of the compound (Table 1) shows the signal of two methyl protons δ H (1.10, CH3-18; 1.02, CH3-19), two oxygen-containing methylene proton signals δ H (4.00 / 3.82, H-20), two oxygenated methine proton signals δ H (3.68, H-1; 3.75, H-6) and an olefin proton signal (δ H 6.24, CH-14). Combined with the carbon spectrum of the compound (Table 1), DEPT spectrum and HSQC spectrum, it was shown that 1 had 17 carbon signals, which can be attributed to: 2 methyl carbon δ C 21.6 (CH3-18), 32.0 (CH3-19); 5 methylene carbons (including 1 oxymethylene) δ C 30.1 (C-2), 40.0 (C-3), 28.0 (C-11), 39.5 (C-12) and 65.1 (C-20); 5 methine carbons (including 2 oxymethylenes and one olefinic methine) δC 73.3 (C-1), 61.1 (C-5), 72.6 (C-6), 50.4 (C-9) and 124.2 (C-14); 5 carbon atoms not connected to hydrogen (including 1 oxygen-connected carbon, 1 olefin carbon and 1 carbonyl carbon) δ C 34.2 (C-4), 96.8 (C-7), 169.0 (C-8), 43.8 (C-10) and 43.8 (C-10). Comprehensive analysis of the NMR data of the compound revealed that it was very similar to the reported compound pharcusin B. The significant feature of compound 1 was the C-1 acetyl group in pharcusin B (δ H 5.10,δc75.6) is replaced by hydroxyl groups (δ H 3.68,δc73.3), 1 H- 1 This inference was confirmed by H COSY and HMBC correlation.

[0055] The relative configuration of the compound was determined by NOESY spectrum analysis. Based on the fact that all the reported enantio-kaurane diterpenoids have the 5β-H configuration characteristics, it is preliminarily speculated that the 5-position hydrogen in compound 1 is in the β orientation. H 1.27) / H-1(δ H 3.68) / H-9(δ H 2.55) / CH3-19(δ H 1.02) and the correlation between H-20(δ H 4.00,3.82) / CH3-18(δ H 1.10) / H-6(δ H 3.75) between the two groups, indicating that H-1, H-5, H-9, and CH3-19 are all β-oriented, while H-20, H-6, and CH3-18 are α-oriented. Experimental determination of the electronic circular dichroism (ECD) curve of compound 1 closely matches the theoretical calculation of the (1S, 5R, 6S, 7S, 9S, 10S)-1 configuration. Therefore, the complete structure of the new compound 1 was determined and named Trichanthone B.

[0056] Table 1 Compound 1 1 H and 13 C NMR data ( 1 H, 500MHz; 13 C,125MHz)

[0057]

[0058]

[0059] Example 2

[0060] The anti-tumor cell activity test study of the present invention is carried out according to the following steps:

[0061] 1. Tumor Cell Culture

[0062] Pancreatic cancer cell line MIA PaCa-2, colon cancer cell line HT-29, lung cancer cell line A549, and liver cancer cell line HepG2 (Cell Bank, Chinese Academy of Sciences) were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum at 37°C and 5% CO2. The culture medium for HT-29, A549, and HepG2 cells was supplemented with 10% FBS and 1% PSN, while the culture medium for MIA PaCa-2 cells was supplemented with 10% FBS, 2.5% HS, and 1% PSN.

[0063] 2. Preparation of Experimental Drugs

[0064] An appropriate amount of Compound 1 prepared in Example 1 was weighed and dissolved in DMSO to a final concentration of 40 mM. The stock solution was stored at 4°C. Prior to the experiment, the stock solution was diluted with DMEM medium, ensuring that the final DMSO concentration was less than 0.1%. Different volumes of DMEM medium were added to dilute the compound to different concentrations. DMEM medium containing 0.1% DMSO was also used as a negative control.

[0065] 3. Drug toxicity to tumor cell lines

[0066] Tumor cells were suspended in culture medium and seeded at a cell density of 8,000 for MIA PaCa-2, HT-29, and HepG2 cells, and 3,000 for A549 cells in 96-well plates (100 μL / well). The cells were cultured at 37°C, 5% CO₂ for 24 hours. During the logarithmic growth phase of the tumor cell lines, various concentrations of the compound (0, 0.004, 0.04, 0.4, 4, and 40 μM) were added and incubated at 37°C, 5% CO₂ for 72 hours.

[0067] 4. CCK-8 assay to detect cell viability

[0068] After the drug interacted with the tumor cells for 72 hours, 10 μL of CCK-8 solution was added to each well and incubated in a sterile incubator for 1 hour. The cells were taken out and the OD value was measured at a wavelength of 450 nm using a microplate reader. The IC value of the compound was calculated using GraphPad Prism 8 software. 50 (half inhibitory concentration), with 5-FU and Carboplatin as positive controls.

[0069] 5. Experimental results are shown in Table 2

[0070] Table 2. Cytotoxic activity of compound 1 (IC 50 :μM)

[0071]

[0072] Experimental conclusion: The cytotoxic activity evaluation of compound 1 revealed that it has significant inhibitory effects on the MIA PaCa-2 pancreatic cancer cell line and the A549 lung cancer cell line. Its effects are stronger than those of the positive drugs 5-Fluorouracil and Carboplatin. The inhibitory effect on the MIA PaCa-2 pancreatic cancer cell line reaches the nanomolar level, and it has the potential to be developed into a new drug for pancreatic cancer and lung cancer.

Claims

1. A novel enantiokaurane compound having antitumor activity, characterized in that: Its structural formula is as follows:

2. The method for preparing the compound according to claim 1, wherein The following steps are involved: (1) Weigh the dried I. trichantha tubers, crush them, add ethanol and water to reflux extraction, filter, collect the filtrate, and concentrate under reduced pressure until there is no alcohol smell to obtain a concentrate; (2) extracting the concentrated solution obtained in step (1) with petroleum ether, dichloromethane, ethyl acetate and n-butanol, respectively, and concentrating under reduced pressure to obtain a petroleum ether fraction, a dichloromethane fraction, an ethyl acetate fraction, an n-butanol fraction and a raffinate fraction, respectively; (3) The dichloromethane fraction obtained in step (2) was subjected to atmospheric pressure column chromatography using AB-8 macroporous resin and eluted with ethanol-water to obtain three sub-fractions DCM-1 to DCM-3; (4) The eluate fraction DCM-3 obtained in step (3) was subjected to silica gel atmospheric pressure column chromatography and eluted with dichloromethane-methanol to obtain 7 sub-fractions DCM-3-1-DCM-3-7; (5) The fraction DCM-3-2 obtained in step (4) was subjected to medium pressure preparative-MCI column chromatography with gradient elution using methanol-water as the mobile phase to obtain 7 fractions DCM-3-2-1-DCM-3-2-7; (6) The fraction DCM-3-2-1 obtained in step (5) was subjected to gel column chromatography gradient elution with methanol-water as the mobile phase to obtain two sub-fractions DCM-3-2-1-1-DCM-3-2-1-2; the fraction DCM-3-2-1-1 was subjected to semi-preparative high performance liquid chromatography to obtain the compound.

3. The method for preparing the compound according to claim 2, wherein The following steps are involved: (1) Weigh the dried tubers of I. trichantha, crush them, and add 95% ethanol by volume to reflux extract the medicinal material. The solid-liquid ratio is 1:6-10. Extract 1-3 times, each time for 1-3 hours. Filter and collect the filtrate. Combine the filtrates and concentrate under reduced pressure until there is no alcohol taste to obtain an extract. (2) The extract obtained in step (1) is made into a suspension with an appropriate amount of water, and then extracted with equal volumes of petroleum ether, dichloromethane, ethyl acetate and n-butanol, each extracted 3 to 6 times, to obtain a petroleum ether fraction, a dichloromethane fraction, an ethyl acetate fraction, an n-butanol fraction and a raffinate water fraction, respectively. (3) The dichloromethane fraction obtained in step (2) was subjected to macroporous resin atmospheric pressure column chromatography, with ethanol-water mobile phases having a volume ratio of 30:70, 60:40, 90:10, and 100:0 as gradient elution, with each gradient elution lasting 3 to 6 column volumes; after thin layer chromatography and ultra-high performance liquid chromatography analysis, the fractions were concentrated and combined using a rotary evaporator to obtain three sub-fractions DCM-1 to DCM-3; (4) The eluate fraction DCM-3 obtained in step (3) was subjected to silica gel atmospheric pressure column chromatography, with dichloromethane-methanol as the mobile phase gradient elution in the following order: 100:0, 50:1, 30:1, 20:1, 10:1, 7:1, 5:1, 3:1, 2:1, 1:1, 1:3, 1:7, 0:100, with each gradient elution lasting 3 to 5 column volumes; after thin layer chromatography and ultra-high performance liquid chromatography analysis, the fractions were concentrated and combined using a rotary evaporator to obtain 7 sub-fractions DCM-3-1-DCM-3-7; (5) The fraction DCM-3-2 obtained in step (4) was subjected to gradient elution on a medium pressure preparative MCI column chromatography with methanol-water as the mobile phase, wherein A is water and B is methanol; the elution gradient program is: 0.01-20.00 min, 25%-25% B; 20.00-50.00 min, 35%-35% B; 50.00-80.00 min, 37%-37% B; 80.00-110.00 min, 38%-38% B; 110.00-140.00 min, 39%-39% B; 140.00 -170.00min, 41%-41% B; 170.00-200.00min, 44%-44% B; 200.00-230.00min, 49%-49% B; 230.00-260.00min, 60%-60% B; 260.00-300.00min, 100%-100% B, elution rate 23mL / min, detection wavelengths 256nm and 310nm; after ultra performance liquid chromatography analysis, the products were concentrated and combined to obtain 7 fractions DCM-3-2-1-DCM-3-2-7; (6) The 150-170 min eluate fraction DCM-3-2-1 obtained in step (5) was subjected to gel column chromatography at atmospheric pressure, eluted with methanol, analyzed by thin layer chromatography and ultra-high performance liquid chromatography, and then concentrated and combined using a rotary evaporator to obtain two sub-fractions DCM-3-2-1-1-DCM-3-2-1-2; (7) The eluate fraction DCM-3-2-1-1 obtained in step (6) was subjected to semi-preparative high performance liquid chromatography to obtain a monomer compound.

4. The method for preparing compound 1 according to claim 3, characterized in that: The specific conditions for ultra-high performance liquid chromatography analysis are as follows: chromatographic column model ACQITY uplc BEH C18 column, size 2.1×100 mm, 1.7 μm, ultra-high performance liquid chromatography column pressure 4135 psi-11328 psi, column temperature 25°C, injection volume 3 μL, 0.1% formic acid water A1-B2 as the mobile phase, elution gradient: 0-10 min, 5%-100% B2, 10-11 min, 100%-5% B2, 11-12 min, 5% B2, flow rate 0.4 mL / min, detection wavelength 190-400 nm.

5. The method for preparing compound 1 according to claim 3, characterized in that: The chromatographic conditions of semi-preparative HPLC were as follows: the chromatographic column model was an X-Bridge C18 preparative column with specifications of 10 nm, 5 μm, 10×250 mm, the HPLC column pressure was 9.8 MPa, the column temperature was 22-26°C, the injection volume was 60 μL, 0.1% formic acid water A-methanol B was used as the mobile phase, the elution was isocratic at a volume ratio of 30:70, the flow rate was 3 mL / min, and the detection wavelengths were 240 nm and 310 nm.

6. Use of the compound according to claim 1 in the preparation of anti-tumor drugs.

7. Use of the compound according to claim 1 in the preparation of drugs for treating pancreatic cancer, colon cancer, lung cancer or liver cancer.

8. A pharmaceutical preparation, characterized in that The compound according to claim 1 and a pharmaceutically acceptable carrier are prepared into a pharmaceutical preparation.

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