CN111303182B - Two novel naphthoquinone dimers in Li medicine red onion and preparation method and application thereof - Google Patents
Two novel naphthoquinone dimers in Li medicine red onion and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to two novel naphthoquinone dimers in Li's medicine red shallot, and a preparation method and application thereof. The compound is obtained by separation through a multistage extraction process, and experiments prove that the two compounds have obvious antibacterial and antitumor activities, and the application prospect in antibacterial and antitumor drugs or health-care products is good.
Description
Technical Field
The invention relates to two novel naphthoquinone dimers in Li medicine red shallots, a preparation method and application thereof, and belongs to the technical field of medical biology.
Background
The herba Alii Fistulosi (Eleutherine americana) is whole plant or bulb of plant of Eleutherine of Iridaceae (Iridaceae), and is also named as herba Alii Fistulosi, and caulis et folium fici Pumilae. According to the records of "Chinese plant journal", about 4 kinds of plants of the genus allium of the family iridaceae are distributed throughout the world, including E.blalbosa, E.angusta, E.citriodora, and E.latifolia, mainly in southeast Asia and American regions. The red onion is bitter in taste and cool in nature, has the effects of clearing heat and removing toxicity, inducing diuresis and removing dampness, removing stasis and reducing swelling, stopping bleeding and the like, can be used as a medicine by bulbs or whole herbs, is widely applied clinically, is collected by Li medicine in China, and is mainly planted and cultivated in Hainan, Guangxi and other areas in China. Due to the problem of sources, the research report of the allium mongolicum regel at home is almost blank at present, while the research of the allium mongolicum regel at home is shallow, and only a small amount of phenolic acid, steroid and a small amount of single quinine are obtained from the allium mongolicum regel. In the aspect of pharmacological activity research, the red onion extract is screened at home to find that the red onion extract has various pharmacological activities including antibacterial, anti-inflammatory, antiviral, antioxidant, cardiovascular and cerebrovascular effects and the like.
Disclosure of Invention
Based on the relatively few researches on new effective components of the red onion in the field, the inventor separates and identifies two novel naphthoquinone dimers from the Li medicament red onion through deep research, and the two novel naphthoquinone dimers are identified and confirmed to have brand-new carbon structure skeletons and have pharmacological activities such as antibiosis, antitumor and the like.
The technical scheme adopted by the invention is as follows:
the invention obtains two novel naphthoquinone dimers in Li medicine red shallots, and the structural formulas are respectively shown as a formula 1 and a formula 2:
the invention also specifically provides a preparation method of the naphthoquinone dimer, which comprises the following steps:
s1: extracting dried scallion bulb powder with methanol, and concentrating the methanol extractive solution under reduced pressure to obtain scallion extract;
s2: dispersing the red onion extract in water, and sequentially extracting with petroleum ether, chloroform, ethyl acetate and n-butanol to obtain different polar parts;
s3: taking the ethyl acetate part, carrying out silica gel column chromatography, taking petroleum ether-ethyl acetate-methanol as an eluent, and carrying out gradient elution to obtain different elution parts;
s4: taking ethyl acetate-methanol parts, carrying out silica gel column chromatography, taking petroleum ether-ethyl acetate as an eluent, carrying out gradient elution, and combining according to Rf value of thin layer chromatography to obtain 8 parts Fr1.A-H; fr1.A developing agent is petroleum ether-ethyl acetate with volume ratio of 1:1, and Rf value is 0.25-0.5; the Fr1.B developing agent is petroleum ether-ethyl acetate with the volume ratio of 1:8, and the Rf value is 0.2-0.4; fr1.C developing agent is petroleum ether-ethyl acetate with volume ratio of 0:1, and Rf value is 0.2-0.5; fr1.D developing agent is ethyl acetate-methanol with volume ratio of 99:1, and Rf value is 0.3-0.6; fr1.E developing agent is ethyl acetate-methanol with volume ratio of 95:5, and Rf value is 0.2-0.5; fr1.F developing agent is ethyl acetate-methanol with volume ratio of 9:1, and Rf value is 0.2-0.5; fr1.G developing agent is ethyl acetate-methanol with volume ratio of 6:1, and Rf value is 0.3-0.6; fr1.H developing agent is ethyl acetate-methanol with volume ratio of 4:1, and Rf value is 0.4-0.6.
S5: performing isocratic elution on Fr1.B by using liquid chromatography and using methanol water as an eluent to obtain a compound 1;
s6: isocratic elution from Fr1.D using liquid chromatography with methanol water as eluent gave compound 2.
Preferably, in the step S1, the feed-liquid ratio of the red onion bulb powder to the methanol is 1kg (8-10) L.
Preferably, in step S3, the elution gradient is: the volume ratio of petroleum ether to ethyl acetate is 10:1, 5:1, 2:1, 1:1 and 0:1, and the volume ratio of ethyl acetate to methanol is 100:1, 80:1, 60:1, 40:1, 30:1, 20:1, 10:1 and 0: 100.
Preferably, in step S4, the ethyl acetate-methanol fraction is eluted with an ethyl acetate-methanol eluent at a volume ratio of 80: 1.
Preferably, in step S4, the elution gradient is: 80:1, 60:1, 40:1, 30:1, 20:1, 10:1, 5:1, 0: 100.
Preferably, in step S5, the volume fraction of methanol-water is 55-60%.
Preferably, in step S6, the volume fraction of methanol-water is 50 to 55%.
According to the invention, the antibacterial activity screening and the anti-tumor activity screening find that the obtained compound 1 and the compound 2 have obvious antibacterial and anti-tumor activities, and the compound has good application prospects in antibacterial and anti-tumor medicines or health-care products.
Compared with the prior art, the invention has the beneficial effects that:
A. the invention obtains two novel naphthoquinone dimers from the red onion, and experiments prove that the compound 1 and the compound 2 can inhibit the growth of various bacteria, wherein the inhibition effect on escherichia coli is most obvious, the MIC values are the lowest and are respectively 0.06 and 0.24 mu g/mL, the naphthoquinone dimer has obvious inhibition activity on methicillin-resistant staphylococcus aureus (MRSA), and the MIC values are respectively 0.78 and 0.32 mu g/mL. The compound 1 and the compound 2 have killing effects on various tumor cells at the concentration of 0.05-0.6 mu M to different degrees and show stronger proliferation inhibition activity, wherein the inhibition rate of the compound 1 on HuH-7, BEL-7402, H22 and S180 reaches more than 50% at the concentration of 0.3 mu M, and the inhibition rate on BEL-7402 and S180 reaches more than 90% at the concentration of 0.6 mu M. The compound 2 has an inhibition rate of more than 50% on HuH-7, S180 and A549 at a concentration of 0.3 mu M, and has an inhibition rate of more than 100% on HuH-7, S180, A549 and AGS at a concentration of 0.6 mu M. The results show that the compound has detailed antibacterial and antitumor activity and has good application prospect in medicaments or health-care products for preventing or treating tumor diseases.
B. The preparation method of the compound is relatively simple, and a high-toxicity reagent is not used in the extraction process, so that the method is safe and efficient.
Drawings
FIG. 1 the major COSY and HMBC correlations of Compounds 1-2
FIG. 2 ECD Spectroscopy of Compounds 1-2
FIG. 3 preparation of Compound 11H-NMR Spectroscopy (DMSO-d)6)
FIG. 4 preparation of Compound 113C-NMR Spectroscopy (DMSO-d)6)
FIG. 5 COSY spectrum of Compound 1 (DMSO-d)6)
FIG. 6 HSQC spectra (DMSO-d) of Compound 16)
FIG. 7 HMBC profile of Compound 1 (DMSO-d)6)
FIG. 8 NOESY spectrum of Compound 1 (DMSO-d)6)
FIG. 9 preparation of Compound 21H-NMR Spectroscopy (CDCl)3)
FIG. 10 preparation of Compound 213C-NMR Spectroscopy (CDCl)3)
FIG. 11 COSY spectrum of Compound 2 (CDCl)3)
FIG. 12 HSQC spectra (CDCl) of Compound 23)
FIG. 13 HMBC spectra (CDCl) of Compound 23)
FIG. 14 NOESY spectrum (CDCl) of Compound 23)
FIG. 15 HR-ESI-MS spectra of Compound 1
FIG. 16 HR-ESI-MS spectra of Compound 2
Detailed Description
The present invention will be described in further detail with reference to the following detailed description and accompanying drawings.
The experimental methods used in the examples of the present invention are all conventional methods unless otherwise specified.
The materials, reagents and the like used in the examples of the present invention can be obtained commercially without specific description.
Example 1:
the preparation method of the naphthoquinone dimer comprises the following steps:
s1: taking dried red onion bulb powder, and leaching for 3 times by using methanol at room temperature, wherein the ratio of material to liquid is 1kg: extracting for 24h each time for 10L, and concentrating the methanol extractive solution under reduced pressure to obtain herba Alii Fistulosi extract;
s2: dispersing the red onion extract in water, and sequentially extracting with petroleum ether, chloroform, ethyl acetate and n-butanol to obtain different polar parts;
s3: taking ethyl acetate parts, loading the ethyl acetate parts by a dry method, loading the ethyl acetate parts by a wet method, carrying out silica gel column chromatography (14 multiplied by 100cm,100 meshes and 200 meshes), and carrying out gradient elution by using petroleum ether-ethyl acetate-methanol as an eluent to obtain different elution parts; the elution gradient was: the volume ratio of petroleum ether to ethyl acetate is 10:1, 5:1, 2:1, 1:1 and 0:1, and the volume ratio of ethyl acetate to methanol is 100:1, 80:1, 60:1, 40:1, 30:1, 20:1, 10:1 and 0: 100;
s4: subjecting ethyl acetate-methanol fraction (80:1) to silica gel column chromatography, eluting with petroleum ether-ethyl acetate (80:1, 60:1, 40:1, 30:1, 20:1, 10:1, 5:1, 0:100) as eluent, gradient eluting, and combining Rf values according to thin layer chromatography to obtain 8 fractions Fr1. A-H; fr1.A developing agent is petroleum ether-ethyl acetate (1:1), and Rf value is 0.25-0.5; fr1.B developing agent is petroleum ether-ethyl acetate (1:8), Rf value is 0.2-0.4; fr1.C developing agent is petroleum ether-ethyl acetate (0:1), Rf value is 0.2-0.5; fr1.D developing agent is ethyl acetate-methanol (99:1), Rf value is 0.3-0.6; fr1.E developing agent is ethyl acetate-methanol (95:5), Rf value is 0.2-0.5; fr1.F developing agent is ethyl acetate-methanol (9:1), Rf value is 0.2-0.5; fr1.G developing agent is ethyl acetate-methanol (6:1), Rf value is 0.3-0.6; fr1.H developing agent is ethyl acetate-methanol (4:1), Rf value is 0.4-0.6;
s5: performing HPLC semi-preparative liquid chromatography on Fr1.B, and isocratically eluting with SB-Phenyl column and 55% methanol water as eluent to obtain compound 1;
s6: and (5) performing HPLC semi-preparative liquid chromatography on Fr1.D by using an SB-Phenyl chromatographic column and using 50% methanol water as an eluent, and isocratically eluting to obtain a compound 2.
Example 2:
the preparation method of the naphthoquinone dimer comprises the following steps:
s1: taking dried red onion bulb powder, and leaching for 3 times by using methanol at room temperature, wherein the ratio of material to liquid is 1kg: extracting for 24h for 8L each time, and concentrating the methanol extractive solution under reduced pressure to obtain herba Alii Fistulosi extract;
s2: dispersing the red onion extract in water, and sequentially extracting with petroleum ether, chloroform, ethyl acetate and n-butanol to obtain different polar parts;
s3: taking ethyl acetate parts, loading the ethyl acetate parts by a dry method, loading the ethyl acetate parts by a wet method, carrying out silica gel column chromatography (14 multiplied by 100cm,100 meshes and 200 meshes), and carrying out gradient elution by using petroleum ether-ethyl acetate-methanol as an eluent to obtain different elution parts; the elution gradient was: the volume ratio of petroleum ether to ethyl acetate is 10:1, 5:1, 2:1, 1:1 and 0:1, and the volume ratio of ethyl acetate to methanol is 100:1, 80:1, 60:1, 40:1, 30:1, 20:1, 10:1 and 0: 100;
s4: subjecting ethyl acetate-methanol fraction (80:1) to silica gel column chromatography, eluting with petroleum ether-ethyl acetate (80:1, 60:1, 40:1, 30:1, 20:1, 10:1, 5:1, 0:100) as eluent, gradient eluting, and combining Rf values according to thin layer chromatography to obtain 8 fractions Fr1. A-H; fr1.A developing agent is petroleum ether-ethyl acetate (1:1), and Rf value is 0.25-0.5; fr1.B developing agent is petroleum ether-ethyl acetate (1:8), Rf value is 0.2-0.4; fr1.C developing agent is petroleum ether-ethyl acetate (0:1), Rf value is 0.2-0.5; fr1.D developing agent is ethyl acetate-methanol (99:1), Rf value is 0.3-0.6; fr1.E developing agent is ethyl acetate-methanol (95:5), Rf value is 0.2-0.5; fr1.F developing agent is ethyl acetate-methanol (9:1), Rf value is 0.2-0.5; fr1.G developing agent is ethyl acetate-methanol (6:1), Rf value is 0.3-0.6; fr1.H developing agent is ethyl acetate-methanol (4:1), Rf value is 0.4-0.6;
s5: performing HPLC semi-preparative liquid chromatography on Fr1.B, and isocratic eluting with SB-Phenyl column and 60% methanol water as eluent to obtain compound 1;
s6: and (5) performing HPLC semi-preparative liquid chromatography on Fr1.D by using an SB-Phenyl chromatographic column and using 55% methanol water as an eluent, and isocratically eluting to obtain a compound 2.
Test example 1: structure identification of compounds
The chemical structure of the obtained active ingredient is determined by applying modern structure identification technology, and the active ingredient with a novel structure is discovered. The results are shown in FIGS. 1-16.
Compound 1:
eleucanasinone A (1) is light yellow powder with molecular formula of C32H28O9(m/z 579.1648[M+Na]+) (calculation value C)32H28NaO9579.1631). The UV spectrum shows an absorption maximum at 213,269,308 nm. The infrared spectrum shows at vmax3450cm-1，νmax1575,1510,1445cm-1，νmax1215,1175,1065cm-1，νmax1705,1655,1610cm-1There is infrared absorption.
1H NMR showed 7 aromatic protic hydrogens, 3 methylene hydrogens, 1 methine hydrogen, 2 methoxy hydrogens and 2 methyl hydrogen signals. 3 aromatic protic hydrogens [ H-7 ] (H7.63,1H,t,J＝8.4Hz),H-8(H7.80,1H,t,J＝8.4Hz),H-9(H7.55,1H,t,J＝8.4Hz)]Is/are as follows1H-1The H coupling constant indicates the presence of an AMX structure in compound 1. Simultaneously, 4 adjacent proton hydrogensH6.68(d,J＝2.4Hz,H-3′),6.28(d,J＝2.4Hz,H-5′),7.11(d,J＝2.4Hz,H-8′)，6.31(d,J＝2.4Hz,H-10′)]Showing 2 AB system structures.13C NMR showed 32 carbon signals in total, including 2 carbonyl carbons ((C-C))C184.0,180.9), 15 quaternary carbons (C: (A)C160.0-71.4), 8 methine carbons: (C118.2,135.2,119.4,111.6,103.8,105.9,99.2), 3 methylene carbons (C: (C)C40.3,28.9,63.4), 2 methoxy groups (A), (B), (C), (D), (C56.3,55.2), 2 methyl carbons: (C24.2,20.4). All hydrogen-carbon signal assignments were obtained by HSQC spectroscopy. The 2D NMR spectrum data are comprehensively analyzed, and the compound contains two structural fragments of 1a and 1 b. 1a has a structure similar to that of the compound eleutherin F, except that the methyl group of C-2 in eleutherin F is at C-2 (C-2: (C-2)C96.2) and C-12(C714) substitution by cyclic ethers with 1 methyl group(s) ((ii)C24.2) is attached to C-12. These structural assignments were confirmed by HMBC spectroscopy. In HMBC spectrogram, H-1(H3.15,d,J＝13.2Hz；H2.29, d, J ═ 13.2Hz) and C-2, C-12, C-13, CH3-13(H1.49, s) and C-12.
The 1b structural fragment was identified as 4 ', 13 ' -dihydroxy-9 ' -methoxy-6 ', 7 ' -benzofurans. In HMBC spectrum, H-3' (H6.68, d, J ═ 2.4Hz) and C-4' ((C-d)C158.2) related, H-5' ((II)H6.28, d, J ═ 2.4Hz) and C-4 ', C-6 ' ((C-4, C-6 ')C154.0) related, H-8' ((II)H7.11, d, J ═ 2.4Hz) and C-7' ((d, J ═ 2.4Hz) andC149.9),C-9′(C160.0) related, H-10' ((II)H6.31, d, J ═ 2.4Hz) and C-9 ', C-12 related, H-13' (C-12) and (C-H4.60,dd,J＝12.0,3.0Hz；H4.73, dd, J ═ 12.0,6.6Hz) and C-1' ((d)C114.5),C-2′(C140.3),C-3′(C111.6) related, OH-4' ((OH-2)H9.74, s) and C-4 'related, OH-13' ((R)H5.47, dd, J ═ 6.6,3.0Hz) and C-13', OCH3-9′(C55.2) and C-9'. Critical related CH313 is related to C-12, C-12 ', H-10' is related to C-12, indicating that the 1b structural fragment is attached at the C-12 position of the 1a structural fragment by a single bond.
The relative configuration of the chiral centers is judged by NOESY spectra and coupling constants. H-4 and H-1a, H-1a and CH313 correlation, indicating that these hydrogens are in the same plane. The ether linkage between C-2 and C-12 is in one orientation. OH-4 ' and OH-13 ', H-3 ', H-5 ' related, H-10 ' and OCH3-9 ', H-8' are related, indicating that they are coplanar. The absolute configuration of the compound cannot be judged by X-ray because of the chemical torsion force caused by the single bond of C-12/C-11' and the prevention of the formation of single crystal of the compound. Therefore, the absolute configuration was measured by Electron Circular Dichroism (ECD), and was judged to be 2S,4S,12R.
From the above analysis, the structure of compound 1 was determined as follows:
compound 2:
eleucanasinone B (2), red brown powder, [ M + H]+m/z 527.1717 (calculated as C)31H27O8527.1706). Compound 2 showed maximum uv absorption at 213,250,269. Infrared absorption peak at vmax3425,1710,1665,1615cm-1。1The H NMR spectrum showed that Compound 2 had an AMX system [ 2 ]H7.45(H-6,1H,d,J＝8.4Hz),H7.53(H-7,1H,t,J＝8.4Hz)，H7.22(H-8,1H,d,J＝8.4Hz)]6 methine hydrogens [ alpha ]H5.14(H-1,q,J＝6.6Hz),H4.49(H-3,m),H4.51(H-4,d,J＝5.4Hz),H6.48(H-1′,s),H7.83(H-5′,s),H6.26(H-10′,s)]4 methyl hydrogen [ alpha ]H1.73(1-CH3,d,J＝6.6Hz),H1.29(3-CH3,d,J＝6.6Hz),H2.96(6′-CH3,s),H2.48(11′-CH3,s)]1 methoxy hydrogen [ alpha ]H3.99(9-OCH3,s)]And 1 hydroxyhydro [ 2 ]H8.55(s)].13C NMR spectrum showed that Compound 2 has 31 carbon atoms including 4 methyl groups, 1 methoxy group, 9 methine groups, 14 quaternary carbons and 3 carbonyl signals (C:)C183.2,183.6,178.9). Hydrocarbon signal attribution was determined by HSQC spectroscopy.
Detailed 1D NMR and 2D NMR analyses showed that the compound consisted of two parts, 2a and 2 b. 2a is a naphthoquinone skeleton, and has a structure similar to that of known compound eleutherin except that CH in 2a is CH3The relative configuration of-1 is the alpha-position. The 2b structural fragment was identified as a naphthalenone backbone, similar in structure to the compound dihydroeleutherinol except for the position of the hydroxyl and methyl groups. In HMBC spectrum, OH-2' (H8.55, s) and C-1' ((C-1)C107.5), C-2 ', C-3' related, CH3-6′(H2.96, s) and C-5' ((S)C122.7)，C-6′(C136.1) depending on the low field mobility of C-7 ' and the molecular weight, OH-7 ' is attached to C-7 ' ((II) ((III))C157.3). H-4 and C-7 ', C-8' ((C-H-C-C111.5), H-3 and C-8 ', indicating that 2b is linked to 2a by a C-8'/C-4 single bond.
The relative configuration of compound 2 was determined by NOESY spectroscopy. CH (CH)3-3 and H-1, H-4 are related, indicating that they are in the same plane. The single crystal cultivation was unsuccessful due to the chemical torsion of the single bond of C-4/C-8'. Therefore, the ECD spectrum is adopted to measure the absolute configuration of the compound 2, and the absolute configuration is shown to be (1S,3S,4R) -2 and is consistent with the ECD experimental spectrum.
From the above analysis, the structure of compound 2 was determined as follows:
TABLE 1 NMR data for Compound 1 (DMSO-d)6)
TABLE 2 NMR data (CDCl) of Compound 23)
Test example 2: screening for antibacterial Activity
2 compounds obtained in example 1 were screened for their activity against methicillin-resistant Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Vibrio cholerae, Klebsiella pneumoniae and Mycobacterium tuberculosis.
Inoculating the strains on culture medium, culturing at 37 deg.C for 24 hr, scraping colonies from the plate, diluting with sterile normal saline, measuring bacterial concentration with Mach's turbidimeter tube, and making into 9 × 106CFU/mL bacterial dilution, 0.05mL bacterial liquid from each bacterial dilution tube, respectively, was spotted on drug plates of various concentrations and drug-free control plates, after incubation at 37 ℃ for 24h, MIC values of the compounds were observed and recorded, and the experiment was repeated 3 times, with the results shown in Table 3.
TABLE 3
The results show that: the compound 1 and the compound 2 can inhibit the growth of various bacteria, wherein the inhibition effect on escherichia coli is the most remarkable, and the MIC values are respectively 0.06 and 0.24 mu g/mL.
Test example 3: screening for antitumor Activity
The MTT method is adopted to research the proliferation inhibition of 2 compounds on Hep G2, SK-HEP-1, HuH-7, BEL-7402, H22, S180, A549, MCF-7, AGS and HCCC-9810, and the growth inhibition effect is evaluated by the inhibition rate. The results are shown in tables 4 and 5.
TABLE 4 tumor inhibition Rate of Compound 1
TABLE 5 tumor inhibition Rate of Compound 2
The results show that: the compound 1 and the compound 2 have killing effects of different degrees on Hep G2, SK-HEP-1, HuH-7, BEL-7402, H22, S180, A549, MCF-7, AGS and HCCC-9810 cell strains under the concentration of 0.05-0.6 mu M, and show stronger proliferation inhibition activity.
The compound 1 has an inhibition rate of more than 50% on HuH-7, BEL-7402, H22 and S180 at a concentration of 0.3 mu M, and has an inhibition rate of more than 90% on BEL-7402 and S180 at a concentration of 0.6 mu M.
The compound 2 has an inhibition rate of more than 50% on HuH-7, S180 and A549 at a concentration of 0.3 mu M, and has an inhibition rate of more than 100% on HuH-7, S180, A549 and AGS at a concentration of 0.6 mu M.
The foregoing is a more detailed description of the present invention that is presented in conjunction with specific embodiments, and the practice of the invention is not to be considered limited to those descriptions. It will be apparent to those skilled in the art that a number of simple derivations or substitutions can be made without departing from the inventive concept.
Claims (10)
1. Two novel naphthoquinone dimers in Li Yao red onion are characterized in that the structural formulas of a compound 1 and a compound 2 are respectively shown as a formula 1 and a formula 2:
2. the method for producing a naphthoquinone dimer according to claim 1, comprising the steps of:
s1: extracting dried scallion bulb powder with methanol, and concentrating the methanol extractive solution under reduced pressure to obtain scallion extract;
s2: dispersing the red onion extract in water, and sequentially extracting with petroleum ether, chloroform, ethyl acetate and n-butanol to obtain different polar parts;
s3: taking the ethyl acetate part, carrying out silica gel column chromatography, taking petroleum ether-ethyl acetate-methanol as an eluent, and carrying out gradient elution to obtain different elution parts;
s4: taking ethyl acetate-methanol parts, carrying out silica gel column chromatography, taking petroleum ether-ethyl acetate as an eluent, carrying out gradient elution, and combining according to Rf value of thin layer chromatography to obtain 8 parts Fr1.A-H;
s5: performing isocratic elution on Fr1.B by using liquid chromatography and using methanol water as an eluent to obtain a compound 1;
s6: isocratic elution from Fr1.D using liquid chromatography with methanol water as eluent gave compound 2.
3. The method for preparing naphthoquinone dimer according to claim 2, wherein in step S1, the ratio of red onion bulb powder to methanol is 1kg (8-10) L.
4. The method for producing the naphthoquinone dimer according to claim 2, wherein in step S3, the elution gradient is: the volume ratio of petroleum ether to ethyl acetate is 10:1, 5:1, 2:1, 1:1 and 0:1, and the volume ratio of ethyl acetate to methanol is 100:1, 80:1, 60:1, 40:1, 30:1, 20:1, 10:1 and 0: 100.
5. The method for producing the naphthoquinone dimer according to claim 2, wherein in step S4, the ethyl acetate-methanol portion is eluted with ethyl acetate-methanol at a volume ratio of 80:1 as an eluent.
6. The method for producing the naphthoquinone dimer according to claim 2, wherein in step S4, the elution gradient is: the volume ratio of petroleum ether to ethyl acetate is 80:1, 60:1, 40:1, 30:1, 20:1, 10:1, 5:1 and 0: 100.
7. The method for producing the naphthoquinone dimer according to claim 2, wherein in step S4, the fr1.a developing solvent is petroleum ether-ethyl acetate in a volume ratio of 1:1, and Rf value is 0.25 to 0.5; the Fr1.B developing agent is petroleum ether-ethyl acetate with the volume ratio of 1:8, and the Rf value is 0.2-0.4; fr1.C developing agent is petroleum ether-ethyl acetate with volume ratio of 0:1, and Rf value is 0.2-0.5; fr1.D developing agent is ethyl acetate-methanol with volume ratio of 99:1, and Rf value is 0.3-0.6; fr1.E developing agent is ethyl acetate-methanol with volume ratio of 95:5, and Rf value is 0.2-0.5; fr1.F developing agent is ethyl acetate-methanol with volume ratio of 9:1, and Rf value is 0.2-0.5; fr1.G developing agent is ethyl acetate-methanol with volume ratio of 6:1, and Rf value is 0.3-0.6; fr1.H developing agent is ethyl acetate-methanol with volume ratio of 4:1, and Rf value is 0.4-0.6.
8. The method for producing the naphthoquinone dimer according to claim 2, wherein in step S5, the volume fraction of methanol and water is 55 to 60%.
9. The method for producing the naphthoquinone dimer according to claim 2, wherein in step S6, the volume fraction of methanol and water is 50 to 55%.
10. The use of the naphthoquinone dimer of claim 1, wherein the use of compound 1 and/or compound 2 in the preparation of a medicament for preventing or treating a neoplastic disease, or in the preparation of a health product for preventing a neoplastic disease.
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