Novel compound with anti-tumor activity, and preparation method and application thereof

Abstract

The preparation method comprises the following steps: deeply researching the chemical components of the root bark of the meliocarpa nudiflora, extracting by adopting ethanol, hydrolyzing the extract by adopting acid, extracting by adopting chloroform, and repeatedly performing column chromatography by adopting silica gel resin, so as to obtain the meliocarpa nudiflora compound with the antitumor activity and the application of the meliocarpa nudiflora compound. And finally, separating and purifying by adopting semi-preparative high performance liquid chromatography to obtain a new compound. An in-vitro anti-tumor inhibition experiment proves that the compound obtained through separation and purification has relatively remarkable anti-tumor activity, particularly has a relatively obvious inhibition effect on myeloma, human cervical cancer and lung adenocarcinoma, is novel in structure and low in cross resistance with existing anti-tumor drugs, and is expected to be developed into an anti-tumor potential drug.

Description

Technical Field

[0001] The invention belongs to the technical field of medicine, and particularly relates to a new compound with anti-tumor activity separated from the bark of the root of Lophatherum truncatum and a preparation method thereof. Background Art

[0002] Meliosma rigida is another name for the plant Meliosma rigida. Its scientific name is Meliosma rigida. It belongs to the genus Meliosma in the family Sabiaceae. Meliosma rigida is an evergreen small tree, about 10 to 20 meters high. The trunk is straight, with gray-brown bark; the young branches are thick and densely covered with rusty soft hairs. The leaves are alternate, leathery, oblanceolate or elliptical, 8 to 25 cm long and 3 to 5 cm wide, with pointed or obtuse tips, cuneate base, entire or sparsely serrated on the upper part, hairy on the upper surface when young, rusty hairy on the lower surface, and the lateral veins are obvious on the back; the petiole is 2.5 to 3 cm long and has short hairs. The panicle is terminal and has rusty soft hairs; the flowers are small and white; there are 5 sepals; 5 petals; and 5 stamens. The drupe is nearly spherical, with an inclined base, a diameter of 5 to 8 mm, and the calyx is persistent.

[0003] The root bark and root bark of the radix strychnifoliae are used as medicine. The root bark is harvested in autumn when it matures and dried. It grows on hillsides, stream banks, forest edges or broad-leaved mixed woods and bushes below 1500m above sea level. It is distributed in Zhejiang, Jiangxi, Fujian, Taiwan, Hubei, Hunan, Guangdong, Guangxi, Guizhou, Yunnan and other places. Root bark: sour, flat. Detoxification, diuresis, swelling. Indications: edema, abdominal distension, nameless swelling, snake bites. Root bark: bitter; flat. Relieves the symptoms and relieves cough. Mainly used for colds and coughs.

[0004] Modern research shows that the schizonepeta has multiple physiological activities such as anti-rheumatic, hypoglycemic, anticonvulsant, sedative, analgesic and antibacterial. At present, there are few studies on the chemical components of the anti-tumor activity of the schizonepeta.

[0005] Previously, the applicant studied the supercritical extract of the fruit of the lycopodii fruit (application number 2023112419621) and found that it has certain anti-tumor activity. This application is based on the existing foundation and conducts a more in-depth study on the active ingredients of its root bark to develop monomer compounds with certain activity, providing a scientific basis for the resource utilization of the lycopodii fruit. Summary of the invention

[0006] The purpose of the present invention is to conduct in-depth research on the active ingredients of the root bark of Psoralea corylifolia, extract and separate the active ingredients with potential activity, and provide a scientific basis for clinical application.

[0007] Technical solution: The present invention separates a new compound with anti-tumor activity from the bark of the plant Pennisetum chinense. The structural formula of the new compound obtained by purification and separation of the present invention is as follows:

[0008]

[0009] Its molecular formula is C 17 H 24 O5.

[0010] A method for preparing a new Meliosma oldhamii compound with anti-tumor activity, which comprises the following steps:

[0011] (1) Weigh the dried root bark of Meliosma oldhamii, crush it appropriately, add a certain amount of ethanol for extraction, combine the extraction solutions, concentrate, add an acid with a certain volume concentration to the concentrate for water bath hydrolysis, after the hydrolysis is completed, filter by suction, wash with water until neutral and then dry to obtain a hydrolysis product. Take the hydrolysis product and add chloroform or ethyl acetate for extraction, combine the filtrates, and concentrate under reduced pressure to obtain a chloroform or ethyl acetate extract;

[0012] (2) Take the chloroform or ethyl acetate extract obtained in step (1), use normal pressure silica gel column chromatography, and elute with petroleum ether - ethyl acetate with different volume ratios as the eluent in gradient; after spotting and analyzing by thin layer chromatography, combine the same elution fractions, concentrate, and obtain 6 elution fractions LF-1 to LF-6;

[0013] (3) After dry loading of the fraction LF-2 obtained in step (2), load it onto a silica gel column chromatography again, and elute with chloroform - methanol with different volume ratios as the eluent in gradient; after spotting and analyzing by thin layer chromatography, combine the same eluents, concentrate, and obtain 4 elution fractions BLZ-1 to BLZ-4;

[0014] (4) Separate and purify the BLZ-4 elution fraction obtained in step (3) by semi-preparative high performance liquid chromatography to obtain a monomeric compound.

[0015] As a preferred embodiment, the method for preparing the new Meliosma oldhamii compound with anti-tumor activity described above comprises the following steps:

[0016] (1) Weigh the dried root bark of Meliosma oldhamii, crush it appropriately, add 6 - 15 times the volume of ethanol, use continuous reflux extraction for 1 - 3 times, each time for 1 - 2 hours, combine the extraction solutions, concentrate, add sulfuric acid with a certain concentration to the concentrate for water bath hydrolysis for 1 - 3 hours, after the hydrolysis is completed, filter by suction, wash with water until neutral and then dry to obtain a hydrolysis product. Take the hydrolysis product and add 6 - 12 times the volume of chloroform for continuous reflux extraction for 1 - 3 times, each time for 0.5 - 2 hours, filter and collect the filtrate, combine the filtrates, and concentrate under reduced pressure to obtain a chloroform extract;

[0017] (2) Take the chloroform extract from step (1), and perform silica gel normal pressure column chromatography. Use petroleum ether - ethyl acetate with volume ratios of 40:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, and 1:4 as eluents for gradient elution, with each gradient eluting for 4 - 6 column volumes; after thin layer chromatography spotting analysis, combine the same eluents, concentrate, and obtain 6 elution fractions LF - 1 to LF - 6;

[0018] (3) Take fraction LF - 2 obtained in step (2), after dry sample loading, perform silica gel column chromatography again. Use chloroform - methanol with volume ratios of 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4, and 1:8 as eluents for gradient elution, with each gradient eluting for 3 - 6 column volumes; after thin layer chromatography spotting analysis, combine the same eluents, concentrate, and obtain 4 elution fractions BLZ - 1 to BLZ - 4;

[0019] (4) Take the elution fraction of BLZ - 4 obtained in step (3), and perform separation and purification using semi - preparative high - performance liquid chromatography. Use acetonitrile (A) - water (B) as the mobile phase for gradient elution, and collect the monomeric compound.

[0020] As a more preferred embodiment, the preparation method of the Melicope patulin compound with anti - tumor activity includes the following steps:

[0021] (1) Weigh dry Melicope patulin root bark, crush it appropriately, add ethanol with a volume 15 times that of the root bark, and perform continuous reflux extraction 3 times. The first extraction is for 2 hours, and the latter two extractions are for 1 hour each. Combine the 3 extraction solutions, concentrate, add sulfuric acid with a volume concentration of 10% in a water bath for hydrolysis for 2 hours according to a volume ratio of 5:1. After hydrolysis, wash with water until neutral and then dry. Take the hydrolysis product, add chloroform with a volume 10 times that of the product, and perform continuous reflux extraction 3 times. The first 2 extractions are for 1 hour each, and the last extraction is for 0.5 hour. Filter and collect the filtrate, combine the filtrates, and concentrate under reduced pressure to obtain the chloroform extract;

[0022] (2) Take the chloroform extract from step (1), and perform silica gel normal pressure column chromatography. Use petroleum ether - ethyl acetate with volume ratios of 40:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, and 1:4 as eluents for gradient elution, with each gradient eluting for 5 column volumes; after thin layer chromatography spotting analysis, combine the same eluents, concentrate, and obtain 6 elution fractions LF - 1 to LF - 6;

[0023] (3) After dry loading the fraction LF-2 obtained in step (2), it is subjected to silica gel column chromatography again, and eluted successively with chloroform-methanol with volume ratios of 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4, and 1:8 as the eluent gradient, with each gradient eluting for 3 column volumes; after thin layer chromatography spotting analysis, the same eluents are combined and concentrated to obtain 4 elution fractions BLZ-1 to BLZ-4;

[0024] (4) The BLZ-4 elution fraction obtained in step (3) is separated and purified by semi-preparative high performance liquid chromatography, and gradient eluted with acetonitrile (A)-water (B) as the mobile phase to collect the monomeric compound.

[0025] As a preferred embodiment, in the preparation method of the new compound of Meliosma rigida Sieb. et Zucc., the chromatographic conditions of the semi-preparative high performance liquid chromatography are: Eclipse XDB Prep C 18 Preparative chromatographic column, specifications 10nm, 5μm, 10×250mm, column temperature 25–30 °C, injection volume 40 μL, with acetonitrile (A)-water (B) as the mobile phase, gradient elution (0-5 min, 80% A, 5–12 min, 40% A, 12–21 min, 20% A, 21–40 min, 45% A), flow rate 3 mL / min, detection wavelength 248 nm, and the monomeric compound is collected in the 21–34 min time period.

[0026] Beneficial effects: The present invention has the following innovative points:

[0027] The present invention deeply studies the chemical constituents of the root bark of Meliosma rigida Sieb. et Zucc., extracts with ethanol, hydrolyzes the glycosidic bond of the extract with acid and then extracts with chloroform, then repeatedly performs column chromatography on silica resin, and finally separates and purifies with semi-preparative high performance liquid chromatography to obtain a new compound. Through in vitro anti-tumor inhibition experiments, it is proved that the compound separated and purified by the present invention has relatively significant anti-tumor activity, especially has obvious inhibitory effects on myeloma, human cervical cancer and lung adenocarcinoma. The compound extracted and separated by the present invention can be prepared into various dosage forms with pharmaceutically acceptable carriers. Description of the drawings

[0028] Figure 1 (+)-HR-ESI-MS diagram of the compound of the present invention.

[0029] Figure 2 The compound of the present invention 1 1H-NMR diagram.

[0030] Figure 3 The compound of the present invention 13 13C-NMR diagram.

[0031] Figure 4 It is the structural formula of the invented compound. Detailed implementation manners

[0032] The present invention can be better understood according to the following embodiments. However, those skilled in the art can easily understand that the specific material ratios, process conditions and their results described in the embodiments are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims. All the materials and reagents for the illustration of the present invention are obtained conventionally in the market.

[0033] Example 1

[0034] 1. Experimental instruments and materials

[0035] Agilent 1260 Infinity II preparative liquid chromatography system, Eclipse XDB Prep C18 preparative chromatographic column (10 nm, 5 μm, 10×250 mm); column chromatography silica gel (200–300 mesh); chromatographic grade acetonitrile was purchased from Merck, USA; analytical grade petroleum ether, chloroform, ethyl acetate, methanol, etc. were purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd.

[0036] 2. The preparation method of the monomer compound comprises the following steps:

[0037] (1) Weigh the dried bark of Meliosma oldhamii, crush it appropriately, add ethanol with a volume 15 times that of the medicinal material, and perform continuous reflux extraction 3 times, 2 hours for the first time, and 1 hour for the latter two times. Combine the 3 extraction solutions, concentrate to obtain a concentrate, add sulfuric acid with a volume concentration of 10% in a water bath for hydrolysis for 2 hours according to a volume ratio of 5:1. After the hydrolysis is completed, filter by suction, wash with water until nearly neutral, and then dry. Take the hydrolysis product and add chloroform with a volume 10 times that of the product for continuous reflux extraction 3 times, 1 hour for the first 2 times, and 0.5 hour for the last time. Filter and collect the filtrate, combine the filtrates, and concentrate under reduced pressure to obtain a chloroform extract;

[0038] (2) Take the chloroform extract obtained in step (1), mix it with silica gel by dry method, and perform silica gel normal pressure column chromatography. Use petroleum ether - ethyl acetate with volume ratios of 40:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4 as the eluent for gradient elution in turn, and elute 5 column volumes for each gradient; after thin layer chromatography spotting analysis, combine the same eluents and concentrate to obtain 6 elution fractions LF-1 to LF-6;

[0039] (3) The fraction LF-2 obtained in step (2) was subjected to dry silica gel loading and then subjected to silica gel column chromatography again. Chloroform-methanol with volume ratios of 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4, and 1:8 was used as the eluent for gradient elution, with 3 column volumes eluted for each gradient; after analysis by thin-layer chromatography spotting, the same eluents were combined and concentrated to obtain 4 elution fractions BLZ-1 to BLZ-4;

[0040] (4) The BLZ-4 elution fraction obtained in step (3) was separated and purified by semi-preparative high-performance liquid chromatography, with acetonitrile (A)-water (B) used as the mobile phase for gradient elution (0 - 5 min, 80% A; 5 - 12 min, 40% A; 12 - 21 min, 20% A; 21 - 40 min, 45% A). The monomeric compound was collected in the time period of 21 - 34 min, and the purity was 99%.

[0041] 3. Structure identification of the compound

[0042] The compound is a white amorphous powder, +40.0 (c 0.01, MeOH). Figure 1 , based on the quasi-molecular ion peak m / z 309.1701 ([M + H] + , C 17 H 25 O5 + calculated value of 309.1704) given by the high-resolution mass spectrum (Waters Synapt G2-Si QTOF) of the compound, the molecular formula of the compound was deduced to be C 17 H 24 O5.

[0043] The following is the structural analysis based on the hydrogen and carbon nuclear magnetic data as shown in Figure 2 and Figure 3 :

[0044] As shown in the structural analysis of the hydrogen spectrum and carbon spectrum in Table 1, the 1 1H-NMR of the compound showed signals of 2 methyl protons at δ H (1.10, CH3-15; 1.02, CH3-16), 1 signal of an oxygenated methylene proton at δ H (4.00 / 3.81, H-17), 2 signals of oxygenated methine protons at δ H (3.68, H-1; 3.75, H-6) and 1 signal of an olefinic proton (δ H 6.24, CH-14). Signals of multiple methylene protons at δ H (1.50, 1.51, 1.52, 1.54, H-3 and H-4), δH (2.09, 2.12, 2.42, 2.53, H-11 and H-12).

[0045] For this compound, 13 the C-NMR and DEPT spectra indicate signals of 17 carbons. Among them, the carbon spectrum shows 2 methyl carbons at δ C 21.6 (CH3-15), 32.0 (CH3-16); 4 methylene carbons, which are δ C 30.1 (C-2), 32.0 (C-3), 28.0 (C-11), 39.5 (C-12) and 1 oxygen-linked methylene carbon at 65.1 (C-17); 3 methine carbons at δ C 73.3 (C-1), 61.1 (C-5), 72.6 (C-6), and an olefinic methine carbon signal at 124.2 (C-14); a carbonyl carbon at 202.8 (C-13).

[0046] By comprehensively analyzing HMBC and NOE to determine the linkage positions and configurations of substituents, it can be speculated that the correlations between H-5 (δ H 1.27) / H-6 (δ H 3.75) / H-9 (δ H 2.55) / CH3-15 (δ H 1.10) and the correlations between H-1 (δ H 3.68) / CH3-16 (δ H 1.02) / H-17 (δ H 4.00, 3.82) indicate that H-5, H-6, H-9, and CH3-15 are all in the β orientation. While H-1, CH3-16, and CH2-17 are in the α orientation. The determined structural formula is as follows:

[0047]

[0048] Table 1 H-NMR and 1 C-NMR data of the compound ( 13 H, 500 MHz; 1 C, 125 MHz, Methanol-d4) 13

[0049]

[0050] Example 2 Experiment on in vitro anti-tumor activity

[0051] 1. Experimental materials

[0052] ​Tumor cell lines: multiple myeloma (MM) cell line ARP-1 cells, lung adenocarcinoma cell line A549, human colon cancer cell SW620, human cervical cancer cell line Hela, human liver cancer cell line SMMC-7721.

[0053] Test drug: the monomeric compound of Melicope pteleifolia prepared in Example 1.

[0054] The 96-well plates were purchased from Costar; RPMI1640 medium was purchased from Gibco; DMEM medium was purchased from Gibco; thiazolyl blue MTT was purchased from Amresco. Newborn bovine serum was purchased from Hangzhou Sijiqing Company.

[0055] The experimental instruments used in the following examples are as follows:

[0056] Experimental instrument: BIO-RAD 680 microplate reader.

[0057] 2. Experimental method

[0058] The present invention adopts the improved MTT method. The multiple myeloma MM cell line ARP-1, lung adenocarcinoma cell line A549, human colon cancer cell SW620, human cervical cancer Hela, and human liver cancer SMMC-7721 were routinely cultured in RPMI-1640 medium containing 10% calf serum, 100 U / mL penicillin, and 0.1 mg / ml streptomycin in an incubator at 37°C and 5% CO2. And digested and subcultured with 0.25% trypsin plus 0.02% EDTA. Then, cells in the logarithmic growth phase were taken. After digestion with trypsin, they were prepared into cell suspensions with RPMI-1640 culture solution containing 10% calf serum to make the cell concentration about 1×10 5 cells / ml. Then they were inoculated into 96-well culture plates, 100 μl of cell suspension was added to each well, and 3 replicate wells were set in each group. Then the test drug (the monomeric compound of Melicope pteleifolia prepared in Example 1) was diluted with incomplete medium to different required gradient concentrations, and 100 μL of the corresponding drug-containing medium was added to each well. At the same time, a negative control group without adding the drug was set up and incubated in an incubator at 37°C and 5% CO2 for 48 h; 5 mg / ml of freshly prepared MTT reagent was added to each well and incubated in an incubator at 37°C and 5% CO2 for another 4 h; centrifuged in a centrifuge, and 150 μL of DMSO was added to each well; then the OD values of each well were measured at a wavelength of 570 nm with a microplate reader. After data processing, with the concentration of each group as the abscissa and the cell survival rate as the ordinate, the growth curve of the cells was plotted, and the IC 50 value of each group of test drugs was calculated using the two-point method (Reed and Muench method).

[0059] 3. Experimental results

[0060] The specific experimental results of the inhibitory rates of the monomeric compounds of the present invention against various tumor cells are shown in Table 1 below.

[0061] Table 1 Inhibitory effects of monomeric compounds on tumor cells (IC 50 value)

[0062] Tumor cell line <![CDATA[IC of the compound 50 (μg / ml)]]> ARP1 8.31 SW620 28.72 Hela 12.90 SMMC-7721 46.27 A549 10.42 。

[0063] The experimental results in Table 1 above show that the new monomeric compounds isolated in the present invention have good anti-proliferative activities against multiple myeloma (MM) cell line ARP-1 cells, human cervical cancer cell line Hela and lung adenocarcinoma cell line A549, and have potential activities for developing anti-tumor agents.

Claims

1. A novel compound of pyrifos with antitumor activity, characterized in that: The structural formula of the compound is as follows:

2. The method for preparing the new pyrocatechol compound with antitumor activity according to claim 1, characterized in that: The following steps are involved: (1) Weighing the dried bark of the root of Lophatherum truncatum, crushing it appropriately, adding a certain amount of ethanol to extract, combining the extracts, concentrating, adding a certain volume concentration of acid to the concentrate for water bath hydrolysis, filtering after the hydrolysis is completed, washing with water until neutral, and then drying to obtain a hydrolyzate, taking the hydrolyzate and adding chloroform or ethyl acetate to extract it, combining the filtrates, and concentrating under reduced pressure to obtain a chloroform or ethyl acetate extract; (2) taking the chloroform or ethyl acetate extract of step (1), applying silica gel atmospheric pressure column chromatography, and gradient eluting with petroleum ether-ethyl acetate in different volume ratios as eluents; after thin layer chromatography spot analysis, combining the same elution fractions, concentrating, and obtaining 6 elution fractions LF-1 to LF-6; (3) The fraction LF-2 obtained in step (2) was dried and then subjected to silica gel column chromatography again, and gradient eluted with chloroform-methanol in different volume ratios as eluents; after thin layer chromatography spot analysis, the same eluents were combined and concentrated to obtain four eluted fractions BLZ-1 to BLZ-4; (4) The BLZ-4 eluate fraction obtained in step (3) is separated and purified by semi-preparative high performance liquid chromatography to obtain a monomer compound.

3. The method for preparing the new compound with anti-tumor activity according to claim 2, characterized in that: The following steps are involved: (1) Weigh the dried bark of the root of Lophatherum truncatum, crush it appropriately, add 6 to 15 times the volume of ethanol, perform continuous reflux extraction for 1 to 3 times, each time for 1 to 2 hours, combine the extracts, concentrate, add a certain concentration of sulfuric acid to the concentrate for hydrolysis in a water bath for 1 to 3 hours, filter it after the hydrolysis is completed, wash it with water until it is neutral, and then dry it to obtain a hydrolyzate, take the hydrolyzate and add 6 to 12 times the volume of chloroform to continuously reflux extract for 1 to 3 times, each time for 0.5 to 2 hours, filter it, collect the filtrate, combine the filtrate, and concentrate it under reduced pressure to obtain a chloroform extract; (2) taking the chloroform extract of step (1), applying silica gel atmospheric pressure column chromatography, and gradient eluting with petroleum ether-ethyl acetate in volume ratios of 40:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4 as eluents, each gradient elution being 4 to 6 column volumes; after thin layer chromatography spot analysis, combining the same eluents and concentrating to obtain 6 elution fractions LF-1 to LF-6; (3) The fraction LF-2 obtained in step (2) was dried and then subjected to silica gel column chromatography again, and gradient eluted with chloroform-methanol in volume ratios of 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4:1:8, with each gradient elution lasting 3 to 6 column volumes; after thin layer chromatography spot analysis, the same eluents were combined and concentrated to obtain four eluted fractions BLZ-1 to BLZ-4; (4) The BLZ-4 eluate fraction obtained in step (3) is separated and purified by semi-preparative high performance liquid chromatography using acetonitrile (A)-water (B) as the mobile phase for gradient elution to collect the monomer compound.

4. The method for preparing the new compound with anti-tumor activity according to claim 3, characterized in that: The following steps are involved: (1) Weigh the dried bark of the root of Lophatherum chinense, crush it appropriately, add 15 times the volume of ethanol, and extract it by continuous reflux for 3 times, the first time for 2 hours, the second time for 1 hour, combine the 3 extracts, concentrate, add 10% volume concentration of sulfuric acid in a volume ratio of 5:1, and hydrolyze it in a water bath for 2 hours. After the hydrolysis is completed, wash it with water until it is neutral and then dry it. Take the hydrolyzate and add 10 times the volume of chloroform to extract it by continuous reflux for 3 times, the first two times for 1 hour, and the last time for 0.5 hour. After filtering, collect the filtrate, combine the filtrates, and concentrate under reduced pressure to obtain a chloroform extract; (2) taking the chloroform extract of step (1), applying silica gel atmospheric pressure column chromatography, and gradient eluting with petroleum ether-ethyl acetate in volume ratios of 40:1, 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4 as eluents, each gradient elution having 5 column volumes; after thin layer chromatography spot analysis, combining the same eluents, concentrating, and obtaining 6 elution fractions LF-1 to LF-6; (3) The fraction LF-2 obtained in step (2) was dried and then subjected to silica gel column chromatography again, and gradient eluted with chloroform-methanol in volume ratios of 20:1, 10:1, 5:1, 3:1, 2:1, 1:1, 1:2, 1:4:1:8, with each gradient elution lasting 3 column volumes; after thin layer chromatography spot analysis, the same eluents were combined and concentrated to obtain 4 eluted fractions BLZ-1 to BLZ-4; (4) The BLZ-4 eluate fraction obtained in step (3) is separated and purified by semi-preparative high performance liquid chromatography using acetonitrile (A)-water (B) as the mobile phase for gradient elution to collect the monomer compound.

5. The method for preparing the pyralidin compound according to any one of claims 2 to 4, characterized in that: The chromatographic conditions for semi-preparative HPLC were: EclipseXDB Prep C 18 A chromatographic column was prepared with specifications of 10 nm, 5 μm, 10×250 mm, column temperature of 25–30 °C, injection volume of 40 μL, acetonitrile (A)-water (B) as the mobile phase, gradient elution (0-5 min, 80% A, 5–12 min, 40% A, 12–21 min, 20% A, 21–40 min, 45% A), flow rate of 3 mL / min, detection wavelength of 248 nm, and monomer compounds were collected in the 21–34 min time period.

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 myeloma, lung adenocarcinoma, colon cancer, cervical cancer or liver cancer.

8. According to the use of claim 6 or 7, the compound and a pharmaceutically acceptable carrier are prepared into a pharmaceutical preparation.

9. The use according to claim 8, wherein the pharmaceutical preparation is a tablet, capsule, granule, pill, injection, powder injection, mixture or oral solution.

10. Use of the compound according to claim 1 in combination with other anti-tumor drugs in the preparation of anti-tumor drugs.

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