Compound isolated from Artemisia argyi and its preparation method and application

Abstract

The present invention belongs to the field of anti-tumor drugs and relates to compounds isolated from Artemisia argyi and their preparation methods and applications. Compounds 1-7 isolated from Artemisia argyi provided in the present invention have the following structural formula: Compounds 1-7 isolated from Artemisia argyi have an inhibitory effect on a variety of cancer cells, and can effectively inhibit the growth of tumor cells at a concentration of 40 μM, and have good practical application value.

Description

Technical Field

[0001] The invention belongs to the field of anti-tumor drugs, and in particular relates to a compound separated from wormwood leaves, and a preparation method and application thereof. Background Art

[0002] The statements in this section merely provide background information related to the present invention and do not necessarily constitute prior art.

[0003] Ovarian cancer (OC) is one of the three major malignant tumors of the female reproductive system, accounting for 2.1% of all cancers. The recurrence rate and mortality rate of ovarian cancer are the highest among female reproductive tract malignant tumors. Cervical cancer (CC), also known as cervical cancer, is one of the most common gynecological malignancies and the fourth leading cause of death from malignant tumors in women. Although clinical treatment techniques such as surgery, radiotherapy, and chemotherapy for ovarian and cervical cancer have made great progress, the incidence of ovarian and cervical cancer in young women continues to increase, seriously threatening women's life and health. Therefore, finding new and effective anti-tumor drugs is one of the research focuses in the field of medicine today. Related studies have found that structural compounds such as sesquiterpenes and flavonoids have different biological activities, such as anti-tumor, anti-inflammatory, anti-malarial, and antibacterial pharmacological effects, and are a group of effective, low-toxic natural small molecules.

[0004] Artemisia argyi Lévl.et.Vant. is a plant of the genus Artemisia in the Asteraceae family. Qi Ai is the wormwood leaf from Qichun County, Hubei Province, and is a famous local medicinal material. Li Shizhen recorded in Compendium of Materia Medica that "in modern times, only the ones from Tangyin are called northern wormwood, and those from Siming are called sea wormwood. Since the Chenghua period, the ones from Qizhou are considered the best and are used as prescriptions. They are valued by the world and are called Qi Ai." Studies have found that Qi Ai contains a variety of chemical components such as terpenes, flavonoids, phenolic acids, polysaccharides and coumarins, among which terpenoid compounds have important physiological activity and medicinal value.

[0005] However, there are few studies on isolating compounds from Artemisia argyi for the prevention or treatment of tumor-related diseases. Summary of the invention

[0006] In order to solve the above problems, the present invention provides a compound separated from wormwood leaves, a preparation method thereof and an application of the compound in the preparation of a method for preventing or treating tumor-related diseases.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] The first aspect of the present invention provides a compound isolated from Artemisia argyi, selected from compounds having the following structural formula:

[0009]

[0010] The second aspect of the present invention provides a method for preparing the above-mentioned compound isolated from wormwood, comprising:

[0011] Heat and extract the mugwort leaves with 60% v / v-80% v / v ethanol solution, and concentrate the extract to obtain a crude extract;

[0012] The crude extract is dissolved in water, extracted with petroleum ether and ethyl acetate in sequence, and the ethyl acetate extract is collected and concentrated to obtain a crude extract;

[0013] The crude extract is separated and purified by column chromatography and high performance liquid chromatography to obtain a compound represented by any one of formulas 1-7.

[0014] In some embodiments, the temperature of the heating extraction is 55-65°C, and the heating extraction time is 8-12h;

[0015] In some embodiments, the heating and leaching is performed 2-4 times.

[0016] In some embodiments, the solid-to-liquid ratio of the wormwood leaf and the ethanol solution is 1:14-20, preferably 1:17.

[0017] Preferably, the concentration of the ethanol solution is 70% v / v.

[0018] In some embodiments, the column chromatography uses one or two of normal phase silica gel column chromatography, reverse phase D101 macroporous resin column chromatography and gel column chromatography.

[0019] In some embodiments, the elution system of the column chromatography includes methanol, dichloromethane-methanol, petroleum ether-ethyl acetate or methanol-water.

[0020] In some embodiments, the specific method of column chromatography is: first, the ethyl acetate extraction part flows through normal phase silica gel column chromatography, and then uses dichloromethane-methanol system to elute at a gradient of 100:0→0:100, and the gradient eluates of each gradient are collected and combined to obtain 7 components Fr.1-7, which are Fr.1, Fr.2, Fr.3, Fr.4, Fr.5, Fr.6, and Fr.7 respectively;

[0021] Fr.6 was subjected to silica gel column chromatography, and eluted with a dichloromethane-methanol system at a gradient of 100:0→0:100 to obtain 6 fractions Fr.6-1 to Fr.6-6; Fr.6-5 was separated by gel column chromatography to obtain 5 fractions Fr.6-5-1 to Fr.6-5-5, and Fr.6-5-1 was purified by HPLC preparative chromatography to obtain the compound shown in Formula 1;

[0022] Alternatively, Fr.7 was subjected to silica gel column chromatography, and eluted with a dichloromethane-methanol system at a gradient ratio of 100:0→0:100 to obtain five fractions Fr.7-1 to Fr.7-5;

[0023] Fr.7-1 was separated by gel column chromatography to obtain three fractions Fr.7-1-1 to Fr.7-1-3. Fr.7-1-1 was recrystallized to obtain the compound shown in Formula 2;

[0024] Alternatively, Fr.7-1-2 is subjected to silica gel column chromatography and eluted with petroleum ether-ethyl acetate to obtain a compound of formula 3;

[0025] Alternatively, Fr.7-1-3 is purified by HPLC preparative chromatography to obtain a compound shown in Formula 4;

[0026] Alternatively, after Fr.7-2 is subjected to gel column chromatography to remove the pigment, it is subjected to silica gel column chromatography and eluted with a petroleum ether-ethyl acetate system at a gradient of 7:1→1:1 to obtain three fractions Fr.7-2-1 to Fr.7-2-3;

[0027] Fr.7-2-1 was purified by HPLC preparative chromatography to obtain the compound shown in formula 5-7.

[0028] In some embodiments, the HPLC uses a reverse phase C18 column, the elution system is methanol-water or acetonitrile-water, the flow rate is 2-3 mL / min, the column temperature is 20-30°C, the injection volume per time is 20-100 μL, and a UV detector is used with detection wavelengths of 210 nm and 254 nm.

[0029] The third aspect of the present invention provides the use of the above-mentioned compound or the compound prepared by the above-mentioned method in the preparation of a drug for preventing or treating tumor-related diseases.

[0030] In some embodiments, the tumor includes: ovarian cancer, cervical cancer;

[0031] In some embodiments, the tumor cells include human ovarian cancer A2780 cells, human ovarian cancer SKOV3 cells, human cervical cancer C-33A cells, and human cervical squamous cell carcinoma SiHa cells.

[0032] Beneficial effects of the present invention

[0033] (1) The present invention provides 7 new compounds isolated from Artemisia argyi, namely 5-hydroxymellein, artecanin, 3-O-methyl-iso-secotanapartholide, 3-methoxytanapartholide, canin, Artanomalide C and guaianolide, whose structures are shown in Formulas 1-7, respectively, which fill the gap in the prior art and further expand the new application of Artemisia argyi.

[0034] (2) The preparation method of the compound provided by the present invention is simple to operate and has good stability; the raw materials are readily available, easy to operate, high in yield, and suitable for industrial production.

[0035] (3) The compound isolated from Artemisia argyi provided by the present invention effectively inhibits the growth of ovarian cancer and cervical cancer cells at 40 μM, and can be used to prepare anti-tumor drugs.

[0036] Advantages of additional aspects of the present invention will be given in part in the following description, and in part will become obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention, and the exemplary embodiments of the present invention and their description are used to explain the present invention and do not constitute improper limitations on the present invention.

[0038] Figure 1 Example 2 Compound 1 1 H-NMR spectrum.

[0039] Figure 2 Example 2 Compound 1 13 C-NMR spectrum.

[0040] Figure 3 For Example 2 Compound 2 13 C-NMR spectrum.

[0041] Figure 4 For Example 2 Compound 3 1 H-NMR spectrum.

[0042] Figure 5 For Example 2 Compound 3 13 C-NMR spectrum.

[0043] Figure 6 For Example 2 Compound 4 13 H-NMR spectrum.

[0044] Figure 7 For Example 2 Compound 413 C-NMR spectrum.

[0045] Figure 8 For Example 2 Compound 5 1 H-NMR spectrum.

[0046] Fig. 9 For Example 2 Compound 5 13 C-NMR spectrum.

[0047] Fig.10 For Example 2 Compound 6 1 H-NMR spectrum.

[0048] Fig.11 For Example 2 Compound 6 13 C-NMR spectrum.

[0049] Fig.12 For Example 2 Compound 7 13 C-NMR spectrum. DETAILED DESCRIPTION

[0050] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0051] The present invention will be further described in detail below in conjunction with specific embodiments. It should be noted that the specific embodiments are intended to explain the present invention rather than to limit it.

[0052] Example 1

[0053] Preparation of compounds

[0054] (1) After washing and drying the fresh mugwort leaves, they were heated and extracted twice at 60°C using 70% ethanol (solid-to-liquid ratio of 1:17), the filtrates were combined, and the extracts were concentrated to obtain a crude extract;

[0055] (2) After the crude extract is fully dissolved in water, it is extracted six times with petroleum ether and ethyl acetate in sequence according to a volume ratio of crude extract aqueous solution to extractant of 1:1, the ethyl acetate extracts are combined, and concentrated to obtain a crude extract (i.e., the crude extract aqueous solution is first extracted multiple times with petroleum ether to remove low polar components; then the crude extract aqueous solution is extracted multiple times with ethyl acetate, and the ethyl acetate extracts are collected);

[0056] (3) The crude fraction extracted with ethyl acetate was subjected to normal phase silica gel (200-300 mesh) column chromatography, and gradient elution was performed with a dichloromethane-methanol system, with the elution gradient being 100:0, 100:1, 80:1, 70:1, 50:1, 30:1, 20:1, 5:1, and 0:100. After TLC thin layer chromatography analysis, it was divided into 7 fractions (Fr. 1-7);

[0057] Fr.6 was subjected to silica gel column chromatography (200-300 mesh) and gradient elution with dichloromethane:methanol (100:0→0:100) to obtain 6 fractions Fr.6-1 to Fr.6-6. Fr.6-5 was separated by gel column chromatography (dichloromethane:methanol 1:1) to obtain 5 fractions Fr.6-5-1 to Fr.6-5-5. Fr.6-5-1 was purified by HPLC preparative chromatography (methanol:water=27:73) to obtain compound 1.

[0058] Fr.7 was subjected to silica gel column chromatography (200-300 mesh) and gradient elution with dichloromethane:methanol (100:0→0:100) to obtain five fractions Fr.7-1 to Fr.7-5;

[0059] Fr.7-1 was separated by gel column chromatography (dichloromethane: methanol 1:1) to obtain three fractions Fr.7-1-1 to Fr.7-1-3. Fr.7-1-1 was recrystallized to obtain compound 2; Fr.7-1-2 was purified by silica gel column chromatography (300-400 mesh) and eluted with petroleum ether: ethyl acetate (3:1) to obtain compound 3; Fr.7-1-3 was purified by HPLC preparative chromatography (acetonitrile: water = 30:70) to obtain compound 4.

[0060] After the pigment of Fr.7-2 was removed by gel column chromatography (dichloromethane: methanol 1:1), it was then chromatographed on a silica gel column (300-400 mesh) and gradient eluted with petroleum ether: ethyl acetate (7:1→1:1) to obtain three fractions Fr.7-2-1 to Fr.7-2-3; Fr.7-2-1 was purified by HPLC preparative chromatography (methanol: water = 45:55) to obtain compounds 5, 6, and 7.

[0061] Example 2

[0062] Compound structure identification

[0063] After all compounds were identified by NMR, the properties and spectral data of the obtained compounds are as follows:

[0064] Compound 1: 5-hydroxymellein, colorless crystals, molecular formula: C 10 H 10 O 4. 1H-NMR (600MHz, Pyr) δ: 7.01 (1H, d, J = 9.0Hz, H-6), 6.69 (1H, d, J = 8.7Hz, H-7), 4.65 (1H,m,H-3),3.39(1H,dd,J=3.4,16.8Hz,H-4b),2.71(1H,m,H-4a),1.39(3H,s,H-9) ; 13C-NMR (126MHz, Pyr) δ: 171.53 (C-1), 156.49 (C-8), 147.91 (C-5), 126.39 (C-10), 125.53 (C-6), 116.95 (C-7), 110.06 (C-11), 77.33 (C-3), 29.95 (C-4), 21.69 (C-9). The above data are consistent with those reported in the literature, so compound 1 is identified as 5-hydroxymellein.

[0065] Compound 2: artecanin, colorless oil, molecular formula C 15 H 18 O 5 . 1H-NMR (600MHz, CDCl3) δ: 6.21 (1H, d, J = 3.5Hz, H-13a), 5.38 (1H, d, J = 3.0Hz, H-13b), 4.47 (1H, dd, J = 10.3, 11.5Hz, H-13b), 3.66 (1H, br s,H-2),3.54(1H,br s, H-3), 1.64 (3H, s, H-15), 1.24 (3H, s, H-14); 13C-NMR (125MHz, CDCl3) δ: 169.7 (C-12), 141.7 (C-11), 118.8 (C-13), 80.8 (C-6), 79.1 (C-1), 73.3 (C-4), 72.3 (C-10), 58.6 (C-2), 58.5 (C-3), 50.3 (C-7), 43.5 (C-5), 33.9 (C-9), 27.5 (C-14), 24.4 (C-8), 19.8 (C-15). The above data are consistent with those reported in the literature, so the compound 2 was identified as artecanin.

[0066] Compound 3: 3-O-methyl-iso-secotanapartholide, colorless oil, molecular formula: C 16 H 20 O 5. 1H-NMR (600MHz, CD3OD) δ: 6.25 (1H, d, J = 2.9 Hz, H-13), 5.76 (1H, d, J = 2.6 Hz, H-13'), 5.09 (1H, d, J = 5.5 Hz, H-6), 4.40 (3H, s, H-16), 3.19 (1H,m,H-3),3.11(1H,m,H-7),2.70(1H,dd,J=7.2,19.0Hz,H-3),2.6(1H,m,H-9),2.45(1H,m,H-2),2.13(1H,s,H-14),2.05(1H,s,H-15) ,1.98(1H,m,H-8');13C-NMR(125MHz,CD3OD)δ:210.30(C-10),205.02(C-1),178.44(C-4),172.71(C-12),140.66(C-5),140.39(C-11),122.77(C-13),77.62(C-3),77.74(C-6),54.59(C-16),43.65(C-7),40.52(C-2),38.30(C-9),31.97(C-14),29.84(C-8),14.16(C-15). The above data are consistent with those reported in the literature, so the compound 3 is identified as 3-O-methyl-iso-secotanapartholide.

[0067] Compound 4: 3-methoxytanapartholide, colorless oil, molecular formula: C 16 H 20 O 5. 1H-NMR (600MHz, CDCl3) δ: 6.35 (1H, d, J = 2.6 Hz, H-13), 5.66 (1H, d, J = 2.6 Hz, H-13'), 4.97 (1H, d, J = 5.0 Hz, H-6), 4.30 (1H, br d,J=6.0Hz,H-3),3.40(3H,s,H-16),3.05(1,m,H-7),2.69(1H,dd,J=6.0,18.0Hz,H-3),2.5(1H,m,H-9),2 .32(1H,dd,J=2.2,18.0Hz,H-2),2.21(1H,s,H-14),2.15(1H,s,H-15),1.8(1H,m,H-8');13C-NMR(125MHz, CDCl3)δ:205(C-1),202.2(C-10),170.2(C-12),138.6(C-11),129.6(C-5),126.4(C-4),121.6(C-13),81.4(C-6),75.2(C-3),58.2(C-16),42.2(C-7),40.1(C-9),38.7(C-2),28.9(C-14),28.7(C-8),13.7(C-15). The above data are consistent with those reported in the literature, so the compound 4 is identified as 3-methoxytanapartholide.

[0068] Compound 5: Canin, colorless oil, molecular formula C 15 H 18 O 5. 1H-NMR (600MHz, CDCl3) δ: 6.21 (1H, d, J = 3.0Hz, H-13b), 5.49 (1H, d, J = 3.0Hz, H-13a), 4.34 (1H, dd, J = 9.5, 11.5Hz, 4.07 (1H, br s, H-3), 3.70 (1H, br s,H-2),3.45(1H,m,H-7),2.63(1H,d,J=11.5Hz,H-5),2.33(1H,m,H-8b),2.10(1H,m,H-8a),2.05(1H,m, H-9b),1.84(1H,m,H-9a),1.57(3H,s,H-15),1.27(3H,s,H-14);13C-NMR(125MHz,CDCl3)d:169.4(C-12) ,139.3(C-11),120.0(C-13),83.3(C-4),79.8(C-6),73.3(C-1),72.2(C-10),64.6(C-2),64.3(C-3),57.8(C-5),45.0(C-7),35.0(C-9),26.5(C-14),23.5(C-8),22.1(C-15). The above data are consistent with those reported in the literature, so the compound 5 is identified as Canin.

[0069] Compound 6: artanomalide C, colorless oil, molecular formula C 15 H 18 O 5 . 1H-NMR (600MHz, CDCl3) δ: 6.21 (2H, d, J = 3.5Hz, H-13), 6.06 (1H, d, J = 1.5Hz, C-3), 5.49 (2H, d, J = 2.9Hz, H-13), 4.4 1(1H,d,J=9.5Hz,H-5),3.41(1H,m,H-7),2.65(1H,s,H-1),2.29(3H,d,J=1.5Hz,H-15),1.8(2H,m,H-8),1.60(3H, s, H-14); 13C-NMR (125MHz, CDCl3) δ: 203.8 (C-2), 177.8 (C-4), 169.5 (C-12), 139.1 (C-11), 131.7 (C-3), 120.1 (C-13), 86.0 (C-6), 82.4 (C-5), 71.6 (C-10), 68.0 (C-1), 41.0 (C-7), 38.6 (C-9), 32.7 (C-14), 22.5 (C-8), 16.2 (C-15). The above data are consistent with those reported in the literature, so compound 6 was identified as artanomalide C.

[0070] Compound 7: guaianolide, colorless oil, molecular formula is C14H17ClO5. 1H-NMR (600MHz, CDCl3) δ: 6.19 (1H, d, J = 3.6 Hz, H-13a), 5.46 (1H, d, J = 3.4 Hz, H-13b), 4.38 (1H, dd, J = 9.6, 11.0 Hz, H-6), 4.10 (1H, d, J = 0.8 Hz, H-4), 3.86 (1H, d, J = 0.8 Hz, H-2), 3.58 (1H, m, H-7), 3.22 (1H, br s,4-OH),2.81(1H,d,J=11Hz,H-5),2.32(1H,m,H-8a),1.91(1H,m,H-9a),1.88(1H,m,H -9b),1.62(1H,m,H-8b),1.57(3H,s,H-14),1.23(3H,s,H-15); 13C-NMR(125MHz, CDCl3) δ: 170.5 (C-12), 140.5 (C-11), 80.0 (C-4), 78.5 (C-6), 73.0 (C-1), 72.0 (C-0), 64.0 (C-3), 63.5 (C-2), 50.0 (C-5), 43.5 (C-7), 33.5 (C-9), 28.0 (C-14), 24.0 (C-15), 22.5 (C-8). The above data are consistent with those reported in the literature, so compound 7 was identified as guaianolide.

[0071] Example 3

[0072] Antitumor activity experiment

[0073] Experimental method: Compound 1-7 obtained in Example 2 and positive drug (paclitaxel) were prepared into 50mM stock solution with DMSO and stored at -20°C. The blank group contained no cells and the sample to be tested, the control well contained cells but no sample to be tested, and the test well contained cells and the sample to be tested. The CCK8 method was used to detect the inhibition rate of the samples on ovarian cancer and cervical cancer cells, and 3 replicate wells were set for each sample concentration.

[0074] Specific operation steps: human ovarian cancer A2780 cells (purchased from Haixing Biotechnology Co., Ltd.) were cultured with RPMI-1640 complete medium (10% FBS, 1% penicillin, 1% streptomycin); human ovarian cancer SKOV3 cells (purchased from Wuhan Pronosai Company) were cultured with McCoy's A complete medium (10% FBS, 1% penicillin, 1% streptomycin); human cervical cancer C-33A cells (purchased from Haixing Biotechnology Co., Ltd.) were cultured with MEM complete medium (10% FBS, 1% penicillin, 1% streptomycin); human cervical squamous cell carcinoma SiHa (purchased from Haixing Biotechnology Co., Ltd.) were cultured with DMEM complete medium (10% FBS, 1% penicillin, 1% streptomycin).

[0075] The cells were placed at 37°C and 5% CO 2 A2780 cells cultured to the logarithmic growth phase were cultured at 5×10 3 100μL cell suspension was evenly spread into a 96-well plate and cultured for 24h until the cells adhered to the wall. Then, 100μL of drug-containing culture medium with a concentration of 40μM was prepared for the test sample, and 200μL of drug-containing culture medium with a concentration of 20μM for the positive drug. The solution was changed and the drug was administered. 24h after administration, the supernatant was discarded, and 100μL of culture medium containing 10% CCK8 was added to each well. After 1.5-2.5h of incubation in the incubator, the absorbance at 450nm was measured in an ELISA instrument.

[0076] A2780 cells, SKOV3 cells, C-33A cells and SiHa cells were treated according to the above method, and the positive drug used was paclitaxel. The results are shown in Table 1.

[0077] Table 1 shows the inhibition results of compounds 1-7 on 4 tumor cell lines at 40 μM.

[0078] Table 1

[0079]

[0080] It can be seen from the data in Table 1 that the compounds 1-7 have high inhibition effects on ovarian cancer cells (A2780, SKOV3).

[0081] It can be seen that compound 1-7 can effectively inhibit the growth of tumor cells at a concentration of 40 μM.

[0082] Experimental Example 4

[0083] Antitumor activity experiment

[0084] Experimental method: The data in Example 3 were screened to select ovarian cancer A2780 and SKOV3 cells that were more sensitive to compounds 1-7. The CCK8 method was used to detect the inhibition rate of the samples on ovarian cancer cells, and 3 replicate wells were set for each sample concentration.

[0085] Specific operation steps: Human ovarian cancer A2780 cells (purchased from Haixing Biotechnology Co., Ltd.) were cultured in RPMI-1640 complete medium (10% FBS, 1% penicillin, 1% streptomycin); human ovarian cancer SKOV3 cells (purchased from Wuhan Pronocell Co., Ltd.) were cultured in McCoy's A complete medium (10% FBS, 1% penicillin, 1% streptomycin). The cells were placed at 37°C and 5% CO 2 A2780 cells cultured to the logarithmic growth phase were cultured at 5×10 3 100 μL of cell suspension was evenly spread into a 96-well plate, and cultured for 24 hours until the cells adhered to the wall and grew. The test samples were respectively prepared with 200 μL of drug-containing culture medium with concentrations of (0.625, 1.25, 2.5, 5, 10, 20 μM), and the positive drug was prepared with 200 μL of drug-containing culture medium with a concentration of 10 μM. The liquid was changed for administration. After 24 hours of administration, the supernatant was discarded, and 100 μL of culture medium containing 10% CCK8 was added to each well. After 1.5-2.5 hours of incubation in the incubator, the absorbance at 450 nm was measured in an ELISA instrument.

[0086] A2780 cells and SKOV3 cells were treated according to the above method. The positive drugs used were paclitaxel, IC 50 represents the cell half inhibitory concentration, and the results are shown in Table 2.

[0087] Table 2

[0088]

[0089] As shown in Table 2, compounds 1-7 have better inhibitory effects on ovarian cancer A2780 and SKOV3 cells.

[0090] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a compound isolated from Artemisia argyi, characterized in that: The compound isolated from Folium Artemisiae Argyi is: ； The preparation method of the compound separated from wormwood leaf comprises: Heat and extract the mugwort leaves with 60% v / v -80% v / v ethanol solution, and concentrate the extract to obtain a crude extract; The crude extract is dissolved in water, extracted with petroleum ether and ethyl acetate in sequence, and the ethyl acetate extract is collected and concentrated to obtain a crude extract; The crude extract is separated and purified by column chromatography and high performance liquid chromatography to obtain a compound represented by formula 7; The column chromatography adopts normal phase silica gel column chromatography; the elution system of the column chromatography is methanol, dichloromethane-methanol, petroleum ether-ethyl acetate or methanol-water; The high performance liquid chromatography uses a reverse phase C18 column, and the elution system is methanol-water or acetonitrile-water; The specific method of the column chromatography is: first, the ethyl acetate extraction part flows through the normal phase silica gel column chromatography, and then the dichloromethane-methanol system is used for gradient elution at 100:0→0:100, and the gradient eluates of each gradient are collected and combined to obtain 7 components Fr.1-7, which are Fr.1, Fr.2, Fr.3, Fr.4, Fr.5, Fr.6, and Fr.7 respectively; Fr.7 was subjected to silica gel column chromatography using a dichloromethane-methanol system at a gradient elution ratio of 100:0→0:100 to obtain five fractions Fr.7-1 to Fr.7-5; After the pigment was removed from Fr.7-2 by gel column chromatography, it was subjected to silica gel column chromatography and eluted with a petroleum ether-ethyl acetate system at a gradient of 7:1→1:1 to obtain three fractions Fr.7-2-1 to Fr.7-2-3; Fr.7-2-1 was purified by HPLC preparative chromatography to obtain the compound shown in Formula 7.

2. The method for preparing a compound isolated from wormwood according to claim 1, characterized in that: The heating extraction temperature is 55-65°C, and the heating extraction time is 8-12 h; Or, the heating and leaching are performed 2-4 times.

3. The method for preparing a compound isolated from wormwood according to claim 1, characterized in that: The solid-to-liquid ratio of the wormwood leaf to the ethanol is 1:14-20; Alternatively, the concentration of the ethanol solution is 70% v / v.

4. The method for preparing a compound isolated from wormwood as claimed in claim 3, characterized in that: The solid-to-liquid ratio of the wormwood leaf and ethanol is 1:

17.

5. The method for preparing a compound isolated from wormwood according to claim 1, characterized in that: The HPLC flow rate was 2-3 mL / min, the column temperature was 20-30 °C, the injection volume was 20-100 μL, and an ultraviolet detector was used with detection wavelengths of 210 nm and 254 nm.

Images