Artemilactone dimer A-G as well as pharmaceutical composition, preparation method and application of Artemilactone dimer A-G

Abstract

The invention provides seven guaiane type sesquiterpene dimer compounds as shown in a structural formula (I), artemisinin dimers A-G (compounds 1-7) as well as a preparation method, a pharmaceutical composition and application of the artemisinin dimers A-G (compounds 1-7), and belongs to the technical field of medicines. The compound has inhibitory activity on human hepatoma cell lines HepG2, Huh7 and SK-Hep-1, can form a pharmaceutical composition with a pharmaceutically acceptable carrier, and can be used for preparing anti-hepatoma drugs.

Description

Technical field:

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to a compound artemisinin dimer AG (1-7), a preparation method and application thereof, a pharmaceutical composition containing the compound and application thereof. Background technology:

[0002] Hepatic carcinoma is the sixth most common cancer and the fourth leading cause of cancer-related death worldwide. It is characterized by high malignancy, rapid progression, and poor prognosis, and its mortality rate is almost close to the incidence rate. Currently, there are 8 drugs used in the clinic to treat hepatic cancer, including 1 VEGFR-2 inhibitor (ramucirumab), 2 PD-1 inhibitors (nivolumab, pembrolizumab), and 5 small molecule tyrosine kinase inhibitors (sorafenib, regorafenib, lenvatinib, cabozantinib, and dorafenib). However, due to their similar structures and targets, drug resistance often occurs. Therefore, it is urgent to find new anti-hepatic cancer drugs with different structures and mechanisms.

[0003] Artemisia myriantha Wall.ex Bessser is also known as Artemisia branch (Sichuan), Artemisia bittersense, and Artemisia nigra (Yunnan). It is mainly distributed in Shanxi (Zhongtiao Mountain), southern Gansu, Qinghai, Sichuan, Guizhou, Yunnan, and Guangxi. Except for Guangxi and southern Yunnan, which are distributed at low altitudes up to 1,000 meters, other provinces are distributed at altitudes of 1,000-2,800 meters. It is commonly found on hillsides, roadsides, and in bushes. It is also distributed in northern India, Bhutan, Nepal, Kashmir, northern Myanmar, and northern Thailand. Artemisia myriantha is used as a folk medicine (Yunnan) with anti-inflammatory effects and is also used as a catalyst for the germination of broad beans. It tastes bitter and is cold in nature. It has the effects of clearing heat, dispelling summer heat, cooling blood, and stopping bleeding. It is mainly used for summer colds, heatstroke fever, bone steaming, hot flashes, vomiting blood, and epistaxis.

[0004] So far, there is no report on the artemisinin dimer AG (1-7) in the existing literature, no report on the pharmaceutical composition with it as an active ingredient, and no report on the use of the pharmaceutical composition in the preparation or treatment of liver cancer drugs. Summary of the invention:

[0005] The purpose of the present invention is to provide a new class of artemisinin dimers AG (1-7) represented by formula (I) with medicinal value and a preparation method and application thereof, a pharmaceutical composition containing artemisinin dimers AG (1-7) and an application thereof. This class of compounds has obvious inhibitory activity against liver cancer cell lines and can be used to prepare anti-liver cancer drugs.

[0006] In order to achieve the above-mentioned object of the present invention, the present invention provides the following technical solutions:

[0007] The present invention provides a series of sesquiterpene dimer compounds, artemisinin dimer AG (1-7), having a structure shown in the following formula (I):

[0008]

[0009] The present invention provides a preparation method of the above-mentioned compound 1-7. The leaves of Artemisia multiflora are dried and crushed, and cold-extracted with 90% ethanol at room temperature for three times, each time for 3 days, and the ethanol extracts are combined. The ethanol is recovered under reduced pressure to obtain 9.1 kg of extract, which is then subjected to silica gel column chromatography, and acetone-petroleum ether is used in a v / v ratio, and gradient elution (0:100, 10:90, 20:80, 30:70, 40:60, 50:50) is used. After TLC development, the fractions are combined to obtain seven fractions Fr.A-Fr.G; the fraction Fr.F is subjected to MCI CHP 20P column (945g, 7×50cm) chromatography, using water-methanol (v / v) and gradient elution (70:30, 50:50, 30:70, 10:90) to obtain five fractions Fr.F-1-Fr.F-5; fraction Fr.F-4 was chromatographed on a silica gel column, using acetic acid acetone-petroleum ether (v / v) and gradient elution (5:95, 10:90, 20:80 and 30:70) to obtain four fractions Fr.F-4a-Fr.F-4d; fraction Fr.F-4b was chromatographed on a silica gel column, using ethyl acetate-petroleum ether (5:95) and isocratic elution to obtain six fractions Fr.F-4b-1-Fr.F-4b-6. Fraction Fr.F-4b-3 was subjected to silica gel column chromatography and isocratically eluted with methanol-chloroform (5:95) to obtain six fractions Fr.F-4b-3a-Fr.F-4b-3f. Fraction Fr.F-4b-3c was subjected to semi-preparative HPLC and eluted with water-acetonitrile (55:45) to obtain compounds 1-3. Fraction Fr.F-4b-3d was subjected to semi-preparative HPLC and eluted with water-acetonitrile (52:48) to obtain compounds 4 and 5. Fraction Fr.F-4b-3e was subjected to semi-preparative HPLC and eluted with water-acetonitrile (64:36) to obtain compounds 6 and 7.

[0010] The present invention provides the use of the above compounds 1-7 in the preparation of anti-liver cancer drugs. The present invention has no particular limitation on the method of the use, and any method known in the art can be used.

[0011] The present invention also provides a pharmaceutical composition, which comprises at least one of the above-mentioned compounds 1-7 of formula (I) and a pharmaceutically acceptable carrier. At the same time, the present invention provides a method for preparing the pharmaceutical composition, wherein the above-mentioned method for preparing the compound obtains the compound 1-7, and at least one of the compounds 1-7 is taken and a pharmaceutically acceptable carrier is added.

[0012] Furthermore, the invention also provides the use of the pharmaceutical composition in preparing anti-liver cancer drugs.

[0013] When at least one of the compounds 1-7 is used to prepare an anti-liver cancer drug, the present invention preferably uses the compound 1-7 directly or in the form of a pharmaceutical composition.

[0014] The pharmaceutical composition provided by the present invention comprises at least one of the above compounds 1-7 in a pharmaceutically acceptable carrier. In the present invention, the pharmaceutically acceptable carrier is preferably a solid, semisolid or liquid diluent, a filler and a pharmaceutical product adjuvant. The present invention has no particular limitation on the pharmaceutically acceptable carrier, and any pharmaceutically acceptable carrier well known in the art, non-toxic and inert to humans and animals can be selected.

[0015] The present invention has no particular limitation on the method for preparing the pharmaceutical composition. At least one of compounds 1-7 can be directly mixed with a pharmaceutically acceptable carrier. The present invention has no particular limitation on the mixing process. A pharmaceutical composition can be obtained by selecting a process well known in the art.

[0016] The present invention provides the use of the pharmaceutical composition of the above technical solution in the preparation of anti-liver cancer drugs. The present invention has no special limitation on the method of the application, and any method well known in the art can be used.

[0017] In the present invention, when the pharmaceutical composition is used to prepare an anti-liver cancer drug, the content of the composition in the drug is preferably 0.1-99%; in the pharmaceutical composition, the content of at least one of the compounds 1-7 in the pharmaceutical composition is preferably 0.5-90%. The pharmaceutical composition of the present invention is preferably used in the form of a dosage per unit body weight. In the present invention, the prepared drug is preferably administered in two forms: injection (intravenous injection, intramuscular injection) and oral administration.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] 1. The present invention provides a series of novel sesquiterpene dimer compounds, artemisia lactone dimer AG (1-7). The present invention found that the ethanol extract of the dry aerial part of Artemisia flos-flora has certain inhibitory activity against three liver cancer cell lines (HepG 2, Huh7 and SK-Hep-1), and further separated six sesquiterpene dimer compounds, artemisia lactone dimer AG (1-7), which have inhibitory activity against three liver cancer cell lines.

[0020] 2. The present invention provides a new method for preparing new compounds 1-7, which has readily available raw materials and is easy to operate.

[0021] 3. The present invention provides a pharmaceutical composition with new compounds 1-7 as active ingredients, providing a new anti-liver cancer drug with better medicinal effects.

[0022] 4. Compounds 1-7 of the present invention have inhibitory activity against three liver cancer cell lines (HepG2, SK-HEP-1 and Huh7). For HepG2 cells, compounds 1 and 4-7 have strong inhibitory activity, and their IC 50 The values ​​ranged from 7.4 to 12.6 μM; compounds 2 and 3 also showed good inhibitory activity, with their IC 50 The values ​​were 17.3 and 16.0 μM, respectively. For Huh7 cells, compounds 4-7 showed comparable inhibitory activity to the positive control sorafenib, with IC 50 The values ​​ranged from 6.9 to 9.7 μM; compounds 1 and 3 showed strong inhibitory activity against Huh7, with IC 50 The values ​​were 18.2 and 13.8 μM, respectively; while compound 2 had a moderate inhibitory activity against Huh7, with an IC 50 The value was 26.5 μM. For SK-Hep-1 cells, compounds 1-2 and 4-7 showed inhibitory activity comparable to that of sorafenib, with IC 50 The values ​​were 11.2, 10.3, 8.3, 9.8, 12.7 and 13.0 μM respectively. The above results indicate that compounds 1-7 isolated from Artemisia annua can be used as drugs to treat liver cancer-related diseases. Description of the drawings:

[0023] Figure 1 It is a schematic diagram of the structural formula of the compound artemisinin dimer AG (1-7) of the present invention. Specific implementation method:

[0024] In order to better understand the essence of the present invention, the following is a description of the artemisinin dimer 1-7 and its preparation method, structural identification, and pharmacological effects in combination with the accompanying drawings using the test examples and embodiments of the present invention, but the present invention is not limited to these test examples and embodiments.

[0025] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Embodiment 1:

[0027] Preparation of artemisinin dimer AG (1-7):

[0028] The leaves of Artemisia multiflora (91 kg) were dried and crushed, and then cold-extracted with 90% ethanol at room temperature for three times, each time for 3 days, and the ethanol extracts were combined. The ethanol was recovered under reduced pressure to obtain 9.1 kg of extract, which was then chromatographed on a silica gel column (27.0 kg, 200-300 mesh, 30×90 cm), using acetone-petroleum ether in v / v, gradient elution (0:100, 10:90, 20:80, 30:70, 40:60, 50:50), and the fractions were combined after TLC development to obtain seven fractions Fr.A-Fr.G; the fraction Fr.F (520 g) was purified by MCI CHP 20P column (945g, 7×50cm) was chromatographed, and water-methanol was used in v / v, and gradient elution was performed (70:30, 50:50, 30:70, 10:90) to obtain five fractions Fr.F-1-Fr.F-5; fraction Fr.F-4 (71.0g) was chromatographed on a silica gel column (700g, 9×25cm), and acetic acid acetone-petroleum ether was used in v / v, and gradient elution was performed (5:95, 10:90, 20:80, 30:70) to obtain four fractions Fr.F-4a-Fr.F-4d; fraction Fr.F-4b (7.2g) was chromatographed on a silica gel column (100g, 3.5×25cm), and ethyl acetate-petroleum ether (5:95) was used in an isocratic manner to obtain six fractions Fr.F-4b-1-Fr.F-4b-6. The fraction Fr.F-4b-3 (2.4 g) was chromatographed on a silica gel column (30 g, 2×25 cm) and eluted isocratically with methanol-chloroform (5:95) to obtain six fractions Fr.F-4b-3a-Fr.F-4b-3f. The fraction Fr.F-4b-3c (240 mg) was subjected to semi-preparative HPLC and eluted with water-acetonitrile (55:45) to obtain compound 1 (8 mg, t R =16.3min), 2(10.1mg,t R =25.9min) and 3 (13mg, t R =24.1min). The fraction Fr.F-4b-3d (190 mg) was subjected to semi-preparative HPLC and eluted with water-acetonitrile (52:48) to give compound 4 (18 mg, t R =27.8min) and 5 (11mg, t R =24.6min). The fraction Fr.F-4b-3e (160 mg) was subjected to semi-preparative HPLC and eluted with water-acetonitrile (64:36) to give compound 6 (12 mg, t R =18.6min) and 7 (11mg, t R =22.7min).

[0029] 1. Instruments and Materials

[0030] The nuclear magnetic resonance spectrum was measured using Avance III 600 (Bruker, Fallanden, Switzerland) with TMS (tetramethylsilane) as the internal standard. The optical rotation was measured using an Autopol VI polarimeter (Rudolph Research Analytical, Hackettstown, USA). The infrared spectrum (IR) was measured using a NICOLET iS10 infrared spectrometer (Thermo Fisher Scientific, Madison, USA) using the KBr pellet method. High-resolution mass spectrometry was measured using a Shimadzu LC-MS-IT-TOF (Shimadzu, Kyoto, Japan). CD and UV spectra were measured using a Chirascan instrument (Applied Photophysics, Surrey, UK). Melting point was measured using X-4B micro melting point instrument was used for determination, and the melting point instrument was purchased from Shanghai Precision Scientific Instrument Co., Ltd. The high performance liquid chromatograph was produced by Shimadzu Corporation, the controller model was CBM-20A, the pump model was LC-20AR, the detector model was SPD-M20A, the column oven model was AT-350, and the chromatographic column model used was Agilent-Eclipse XDB-C20 (5μm, 9.4×250mm). The medium pressure liquid phase was a product of Beijing Innovation Hengtong Company, and the MCI column was a product of Mitsubishi Corporation of Japan, model CHP-20P (75~150μm). Thin layer chromatography silica gel plate HSGF254 and column chromatography silica gel (200~300 mesh) were purchased from Yantai Xinnuo Chemical Co., Ltd.; column chromatography dextran gel LH-20 was purchased from GE Healthcare Bio-Sciences AB. Chromatographically pure acetonitrile and methanol were purchased from Merida (Beijing, China), and deionized channel water was purified by MingCheTM-D24UV Merk Millipore system. Analytical pure methanol and acetonitrile were purchased from Tianjin Damao Chemical Reagent Factory. The color developing agent was 10% H2SO4-EtOH solution.

[0031] 2. Structural data of compounds 1-7:

[0032]

[0033] Artemisinin dimer A(1)

[0034] Molecular formula: C 30 H 36 O7

[0035] Molecular weight: 508.25

[0036] Appearance: White powder

[0037] Optical rotation: (c 0.057, methanol)

[0038] HRESIMS (-) m / z: Observed value 553.2437 [M+HCOO] - , calculated value 553.2442 [M+HCOO] - .

[0039] IR(KBr)ν max :3450,1763,1634,1606,1511,1435cm -1 .

[0040] ECD(Methanol)λ max (Δε):216(+29.6),277(+0.77),245(-8.46)nm.

[0041] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0042]

[0043] Artemisinin dimer B(2)

[0044] Molecular formula: C 30 H 36 O8

[0045] Molecular weight: 524.24

[0046] Appearance: White powder

[0047] Optical rotation: (c 0.052, methanol)

[0048] HRESIMS (+) m / z: Observed value 525.2464 [M+H] + , calculated value 525.2483[M+H] + IR(KBr)ν max :3444,1756,1633,1444,1383cm -1 .

[0049] ECD(Methanol)λ max (Δε):200(+8.55),229(-1.11)nm.

[0050] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0051]

[0052] Artemisia lactone dimer C(3)

[0053] Molecular formula: C 30 H 38 O7

[0054] Molecular weight: 510.26

[0055] Appearance: White powder

[0056] Optical rotation: (c 0.110, methanol)

[0057] HRESIMS (+) m / z: Observed value 533.2493 [M+Na] + , calculated value 533.2510 [M+Na] + IR(KBr)ν max :3454,1746,1638,1453,1406cm -1 .

[0058] ECD(Methanol)λ max (Δε):232(+1.11),211(-9.61)nm.

[0059] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0060]

[0061] Artemisia lactone dimer D(4)

[0062] Molecular formula: C 30 H 38 O7

[0063] Molecular weight: 510.26

[0064] Appearance: White powder

[0065] Optical rotation: (c 0.040, methanol)

[0066] HRESIMS (+) m / z: Observed value 533.2521 ([M+Na] + , calculated value 533.2510 [M+Na] + ; IR(KBr)ν max :3448,1745,1640,1458,1381cm -1 .

[0067] ECD(Methanol)λ max (Δε):201(+11.2),226(-1.40)nm.

[0068] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0069]

[0070] Artemisinin dimer E(5)

[0071] Molecular formula: C 30 H 34 O7

[0072] Molecular weight: 506.23

[0073] Appearance: White powder

[0074] Optical rotation: (c 0.050, methanol)

[0075] HRESIMS (+) m / z: Observed value 507.2353 [M+H] + , calculated value 507.2377[M+H] + IR(KBr)ν max :3449,1752,1685,1631,1459,1379cm -1 .

[0076] ECD(Methanol)λ max (Δε):212(+3.35),299(+0.36),231(-2.02),266(-4.49)nm. 1 HNMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0077]

[0078] Artemisia lactone dimer F(6)

[0079] Molecular formula: C 30 H 34 O7

[0080] Molecular weight: 506.23

[0081] Optical rotation: (c 0.090, methanol)

[0082] HRESIMS (-) m / z: Observed value 551.2301 [M+HCOO] -, calculated value 551.2287[M+HCOO] - .

[0083] IR(KBr)ν max :3438,1769,1734,1637,1449cm -1 .

[0084] ECD(Methanol)λ max (Δε):201(+14.8),227(-1.24)nm.

[0085] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0086]

[0087] Artemisia lactone dimer G (7)

[0088] Molecular formula: C 30 H 34 O7

[0089] Molecular weight: 506.23

[0090] Optical rotation: (c 0.055, methanol)

[0091] HRESIMS (-) m / z: Observed value 551.2269 [M+HCOO] - , calculated value 551.2287[M+HCOO] - .

[0092] IR(KBr)ν max :3445,1768,1711,1648,1449,1383cm -1 .

[0093] ECD(Methanol)λ max (Δε):200(+17.47),230(+2.77),216(-1.05)nm.

[0094] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 1 and 2.

[0095]

[0096] Table 2. Compounds 1-7 (δ in ppm) 13 C NMR (150 MHz) data

[0097]

[0098] a CDCl3 was used as solvent. b Use C5D5N as solvent

[0099] Embodiment 2:

[0100] Inhibitory activity of compounds 1-7 against three liver cancer cell lines.

[0101] 1. Materials and Methods

[0102] 1.1 Materials

[0103] HepG2, Huh7, and SK-Hep-1 cell lines were purchased from Shanghai Jining Biotechnology Co., Ltd.; culture medium and serum (EME and FBS) were purchased from Shanghai Longtian Biotechnology Co., Ltd.; MTT was purchased from Guizhou Saiguo Biotechnology Co., Ltd.; Sorafenib was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; DMSO was purchased from Soleberg Technology (Beijing, China)

[0104] 1.2 Instruments

[0105] Flex Station 3 multifunctional microplate reader (Bio-RAD 680, USA); analytical balance (AG135, Metler Toledo, China); constant temperature box (DHP-9082, Shanghai).

[0106] 1.3 Experimental process

[0107] 1). Take the hepatoma cells in logarithmic phase, discard the old culture medium, wash twice with PBS, and discard the PBS;

[0108] 2) Digest the cells with 0.25% trypsin. When the cell outline becomes darker and tends to become round under the microscope, quickly remove the trypsin.

[0109] 3) Use DMEM complete medium containing 10% FBS to terminate digestion and resuspend the cells. Take 10 μL of the cell suspension, count it with a cell counter, and adjust the cell concentration to 1×10 4 / mL, inoculated on a 96-well plate, 100 μL of cell suspension was added to each well, and incubated in a 37°C, 5% CO2 incubator for 24 h to allow the cells to adhere;

[0110] 4). Aspirate the culture medium and add the diluted sample to the plate, add 100 μL to each well, set up 3 replicate wells for each concentration, and continue incubating in the incubator for 48 hours;

[0111] 5). Aspirate the culture medium, add the prepared MTT solution (1 mg / mL), add 100 μL to each well, and incubate in the incubator for 4 hours;

[0112] 6). Aspirate the MTT solution, add DMSO, add 100 μL to each well, and incubate in the incubator for 10 minutes;

[0113] 7). Use an ELISA reader to measure the absorbance at 490 nm, and calculate the cell inhibition rate using the formula: inhibition rate = (negative - experimental group) / (negative - blank group) × 100%, and use the statistical software GraphPad prism 5 to calculate IC 50 , the experiment was repeated 3 times.

[0114] 2. Results

[0115] The inhibitory activities of compounds 1-7 against three liver cancer cell lines (HepG2, Huh7 and SK-Hep-1) are shown in Table 3. For HepG2 cells, compounds 1 and 4-7 have strong inhibitory activities, and their IC 50 The values ​​were 9.8, 9.2, 7.4, 11.4 and 12.6 μM, respectively.

[0116] For Huh7 cells, compounds 4-7 showed stronger or equivalent inhibitory activity to the positive drug sorafenib, with IC 50 The values ​​were 6.9, 9.7, 7.9 and 8.6 μM, respectively; the inhibitory activity of compounds 1 and 3 on Huh-7 cells, their IC 50 The values ​​were 18.2 and 13.8 μM, respectively.

[0117] Compounds 1-2 and 4-7 showed strong inhibitory activity against SK-Hep-1 cells, with IC 50 The values ​​are 11.2, 10.3, 8.3, 9.8, 12.7 and 13.0 μM, respectively.

[0118] Table 3 Inhibitory activity of compounds 1-7 on three liver cancer cell lines

[0119]

[0120] a The value is expressed as IC 50 ±SD

[0121] 3. Conclusion

[0122] Based on the above experimental results, it can be concluded that compounds 1-7 showed certain inhibitory activity against three liver cancer cell lines (HepG2, Huh7 and SK-Hep-1). For HepG2 cells, compounds 1 and 4-7 had strong inhibitory activity, with IC 50The values ​​ranged from 7.4 to 12.6 μM, among which compound 5 had an inhibitory activity comparable to that of the positive drug sorafenib, with an IC 50 The value was 7.7 μM. For Huh7 cells, compounds 4-7 showed stronger or equivalent inhibitory activity than sorafenib, and their IC 50 The values ​​ranged from 6.9 to 9.7 μM. For SK-Hep-1 cells, compounds 1-2 and 4-5 showed inhibitory activity comparable to that of the positive drug sorafenib, with IC 50 The values ​​were 11.2, 10.3, 8.3 and 9.8 μM, respectively. The above results indicate that compounds 1-7 isolated from Artemisia leucophylla can be used as drugs for liver cancer-related diseases.

[0123] Preparation Example:

[0124] In the following formulation examples, conventional reagents are selected and the formulations are prepared according to conventional methods. This formulation example only reflects that at least one of the compounds 1-7 described in the present invention can be prepared into different formulations, and the specific reagents and operations are not specifically limited:

[0125] 1. Dissolve at least one of the compounds 1-7 of the present invention in DMSO, add water for injection according to conventional methods, filter, and sterilize by filling to prepare an injection solution, wherein the concentration of the injection solution is 0.5-5 mg / mL.

[0126] 2. Dissolve at least one of the compounds 1-7 of the present invention in DMSO, dissolve it in sterile water for injection, stir to dissolve it, filter it with a sterile suction funnel, filter it with sterile fine filtration, pack it into ampoules, freeze-dry it at low temperature, and seal it with sterile sealing to obtain a powder injection.

[0127] 3. Add at least one of the compounds 1-7 of the present invention to an excipient at a mass ratio of 9:1 to the excipient to prepare a powder.

[0128] 4. Add at least one of the compounds 1-7 of the present invention to the excipient at a mass ratio of 5:1, and granulate and tablet.

[0129] 5. Prepare at least one of the compounds 1-7 of the present invention into an oral liquid according to a conventional oral liquid preparation method.

[0130] 6. At least one of the compounds 1-7 of the present invention is added to an excipient at a mass ratio of 5:1 to the excipient to prepare a capsule.

[0131] 7. At least one of the compounds 1-7 of the present invention is added to an excipient at a mass ratio of 5:1 to the excipient to prepare granules.

[0132] It can be seen from the above embodiments that the present invention provides a compound in Artemisia polyanthus and its preparation method and application, a pharmaceutical composition and its application. The Artemisia polyanthus lactone dimer provided by the present invention mainly includes 7 compounds with novel structures, which have different degrees of inhibitory activity on liver cancer cells, can be combined with pharmaceutically acceptable carriers to form a pharmaceutical composition, and can be used to prepare anti-liver cancer drugs.

[0133] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. The compound represented by the structural formula (I) is artemisinin dimer AG, namely compound 1-7, 2. The method for preparing the compound of formula (I) artemisinin dimer AG, i.e. compound 1-7, according to claim 1, characterized in that: The method comprises the following steps: drying and crushing the leaves of Artemisia multiflora, cold soaking and extracting them three times with 90% ethanol at room temperature, each time for 3 days, combining the ethanol extracts, recovering the ethanol under reduced pressure to obtain an extract, and then performing silica gel column chromatography, using acetone-petroleum ether gradient elution with a v / v ratio of 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, and combining the fractions after TLC development to obtain seven fractions Fr.A-Fr.G; the fraction Fr.F is subjected to MCI CHP 20P column chromatography, using v / v 70:30, 50:50, 30:70, 10:90 water-methanol gradient elution to obtain five fractions Fr.F-1-Fr.F-5; fraction Fr.F-4 was subjected to silica gel column chromatography, using v / v 5:95, 10:90, 20:80 and 30:70 acetone-petroleum ether gradient elution to obtain four fractions Fr.F-4a-Fr.F-4d; fraction Fr.F-4b was subjected to silica gel column chromatography, using 5:95 ethyl acetate-petroleum ether elution to obtain six fractions Fr.F-4b-1-Fr.F-4 b-6; fraction Fr.F-4b-3 was subjected to silica gel column chromatography and eluted with 5:95 methanol-chloroform to obtain six fractions Fr.F-4b-3a-Fr.F-4b-3f; fraction Fr.F-4b-3c was subjected to semi-preparative HPLC and eluted with 55:45 water-acetonitrile to obtain compounds 1-3; fraction Fr.F-4b-3d was subjected to semi-preparative HPLC and eluted with 52:48 water-acetonitrile to obtain compounds 4 and 5; fraction Fr.F-4b-3e was subjected to semi-preparative HPLC and eluted with 64:36 water-acetonitrile to obtain compounds 6 and 7.

3. Use of the compound of formula (I) artemisinin dimer AG, i.e. compound 1-7, as claimed in claim 1 in the preparation of anti-liver cancer drugs.

4. A pharmaceutical composition comprising at least one of the compounds of formula (I) artemisinin dimer AG, i.e., compounds 1-7, as described in claim 1 and a pharmaceutically acceptable carrier.

5. Use of the pharmaceutical composition according to claim 4 in the preparation of anti-liver cancer drugs.

6. The method for preparing the pharmaceutical composition according to claim 4, characterized in that: The method comprises the following steps: drying and crushing the leaves of Artemisia multiflora, cold soaking and extracting them three times with 90% ethanol at room temperature, each time for 3 days, combining the ethanol extracts, recovering the ethanol under reduced pressure to obtain an extract, and then performing silica gel column chromatography, using acetone-petroleum ether gradient elution with a v / v ratio of 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, and combining the fractions after TLC development to obtain seven fractions Fr.A-Fr.G; the fraction Fr.F is subjected to MCI CHP 20P column chromatography, using v / v 70:30, 50:50, 30:70, 10:90 water-methanol gradient elution to obtain five fractions Fr.F-1-Fr.F-5; fraction Fr.F-4 was subjected to silica gel column chromatography, using v / v 5:95, 10:90, 20:80 and 30:70 acetone-petroleum ether gradient elution to obtain four fractions Fr.F-4a-Fr.F-4d; fraction Fr.F-4b was subjected to silica gel column chromatography, using 5:95 ethyl acetate-petroleum ether elution to obtain six fractions Fr.F-4b-1-Fr.F-4b-6; fraction Fr.F-4b-3 After silica gel column chromatography, eluted with 5:95 methanol-chloroform to obtain six fractions Fr.F-4b-3a-Fr.F-4b-3f; fraction Fr.F-4b-3c was subjected to semi-preparative HPLC and eluted with 55:45 water-acetonitrile to obtain compounds 1-3; fraction Fr.F-4b-3d was subjected to semi-preparative HPLC and eluted with 52:48 water-acetonitrile to obtain compounds 4 and 5; fraction Fr.F-4b-3e was subjected to semi-preparative HPLC and eluted with 64:36 water-acetonitrile to obtain compounds 6 and 7; any one of compounds 1-7 or any combination thereof was added with a pharmaceutically acceptable carrier.

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