Sesquiterpene trimer from Artemisia macrocarpa and its pharmaceutical composition and its preparation method and application

Abstract

The present invention provides a sesquiterpene trimer in Artemisia grandis and a pharmaceutical composition thereof and a preparation method and application thereof, belonging to the field of pharmaceutical technology. The present invention provides 8 new sesquiterpene trimers shown in structural formula (I), Artemsieverolactones A-H (1-8) have obvious cytotoxic activity to human hepatic stellate cells (HSC-LX2), can be combined with a pharmaceutically acceptable carrier or excipient to form a pharmaceutical composition, and can be used to prepare an anti-liver fibrosis drug.

Description

Technical field:

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to a sesquiterpene trimer compound artemsieverolactones AH (1-8), a preparation method thereof, a pharmaceutical composition containing the compound as an active ingredient, and application thereof in pharmaceutical manufacturing. Background technology:

[0002] Epidemiological survey results show that the average number of deaths from liver diseases worldwide is about 2 million per year, accounting for about 3.5% of the world's deaths, and has become one of the main causes of human death. Hepatic fibrosis (HF) is a common pathological process of liver damage caused by multiple causes (viral hepatitis, non-alcoholic fatty hepatitis, cholestasis, alcoholism or autoimmune diseases) and chronic liver diseases caused by inflammation. Liver fibrosis is a dynamic process, specifically manifested as abnormal activation of hepatic stellate cells (HSC), abnormal proliferation of fibrous tissue in the liver, and excessive deposition of extracellular matrix (ECM), which in turn affects the normal physiological structure and function of the human liver. If this process is not inhibited, it may further develop into cirrhosis or liver cancer. Liver fibrosis is a common pathological process of most liver diseases, that is, excessive deposition of extracellular matrix in the liver occurs under the action of various pathogenic factors, thereby affecting the physiological structure and function of the liver. If it is not inhibited or reversed in time, it will accelerate the occurrence of cirrhosis or liver cancer, seriously affecting the quality of life of the human body. At present, single control of etiology or traditional Chinese medicine liver protection is the main way to treat liver fibrosis. There is no chemical synthetic drug for the treatment of liver fibrosis in clinical practice. Although the research and development of anti-fibrosis drugs has made certain progress in recent years, some candidate drugs are difficult to achieve the expected therapeutic effect or have side effects in clinical trials. Hepatic stellate cells are the main source of myofibroblasts. In normal liver, HSC proliferates very slowly, and maintains its dynamic balance by regulating the synthesis and degradation of extracellular matrix. When the liver is continuously stimulated by external pathogenic factors, a series of complex mechanisms are activated, and the final result is that HSC is transformed from a quiescent type to an activated type, and secretes various profibrogenic factors such as TGF-β and PDGF, producing a large amount of disorderly accumulated extracellular matrix (ECM), which promotes the occurrence and development of fibrosis. Therefore, HSC is a potential target for anti-liver fibrosis treatment. Currently, many anti-liver fibrosis drugs under development aim to achieve the therapeutic purpose by inhibiting the proliferation and fibrogenic ability of HSC and promoting the apoptosis of HSC. The present invention aims to find new anti-liver fibrosis active ingredients from natural products based on a human hepatic stellate cells line LX2 (HSC-LX2) screening model.

[0003] Artemisia sieversiana is a perennial herbaceous plant of the genus Artemisia in the Asteraceae family. It grows on hillsides, grasslands, sparse forests and forest edges below 3,500 meters above sea level. It is mainly distributed in Heilongjiang, Liaoning, Inner Mongolia, Gansu, Sichuan, Yunnan, Tibet and other places. According to the "Chinese Medicine Dictionary", Artemisia sieversiana is sweet and bitter in nature, and is mainly used to treat wind-cold, dysentery, jaundice, etc. Mongolian and Tibetan medicines record that it has the effects of reducing swelling, stopping bleeding, clearing heat and relieving summer heat. It can be used to treat diarrhea, colds, jaundice, etc. after being decocted in water, and can be used to treat skin burns caused by ultraviolet radiation. Modern pharmacological studies have shown that Artemisia sieversiana has significant expectorant and antiasthmatic, anti-inflammatory and antibacterial effects. Literature research found that the main components of Artemisia sieversiana are sesquiterpenes (guaiacane type, gemmarane type, muroinane type, sesquiterpenoid dimer), lignans, flavonoids, volatile oils and other types of compounds. Pharmacological studies have shown that the compounds isolated from Artemisia annua have anti-tumor, anti-inflammatory, antibacterial and antioxidant activities.

[0004] So far, there is no report on artemsieverolactones AH (1-8) in the prior art, nor is there any report on compounds 1-8 and their pharmaceutical compositions as anti-liver fibrosis drugs. Summary of the invention:

[0005] The purpose of the present invention is to provide a new class of sesquiterpene trimers with medicinal value, artemsieverolactones AH (1-8), and a preparation method and application thereof, a pharmaceutical composition containing artemsieverolactones AH and an application thereof. In the preliminary work of the present invention, it was found that the Artemisia macrophylla ethanol extract has significant HSC-LX2 cytotoxic activity (the inhibition rate is 58.0% when the concentration is 100 μg / mL), and 8 new sesquiterpene trimers, artemsieverolactones AH (1-8), are obtained by activity-oriented separation. This class of compounds has obvious cytotoxic activity on hepatic stellate cells and can be used to prepare anti-liver fibrosis 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 trimer compounds, artemsieverolactones AH (1-8), having the structure shown in the following formula (I):

[0008]

[0009] The present invention provides a preparation method of the above-mentioned sesquiterpene trimer compounds 1-8. The dried aerial part of Artemisia grandis is crushed, and cold-extracted with 95% ethanol twice, each time for 4 days, the ethanol extract is combined, and the ethanol extract is recovered under reduced pressure. The extract is dispersed in water and extracted with ethyl acetate, and then concentrated to obtain an ethyl acetate extraction part, and then the ethyl acetate extraction part is subjected to silica gel column chromatography, and acetone-petroleum ether is calculated by v / v, 5:95, 10:90, 20:80, 3 0:70 and 100:0 as eluent, gradient elution gave five fractions Fr.A-Fr.E (149g, 101g, 98g, 153g and 164g); fraction Fr.D was eluted by silica gel column chromatography with methanol-chloroform (v / v) at 5:95, 10:90 and 20:80 to give five fractions Fr.D1-Fr.D5 (25g, 10g, 60g, 30g and 25g); fraction Fr.D4 (30g) was chromatographed on an MCI gel CHP 20P column, using water-methanol at 10:90 and 0:100 gradient elution and semi-preparative HPLC to give compounds 1 (22mg), 2 (4mg), 3 (9mg), 4 (10mg), 5 (5mg), 6 (6mg), 7 (3mg), 8 (5mg).

[0010] The present invention provides the use of the sesquiterpene trimer compounds 1-8 described in the above technical solution in the preparation of anti-liver fibrosis 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.

[0011] The present invention provides a pharmaceutical composition, which comprises at least one of the sesquiterpene trimer compounds 1-8 described in the above technical solution and a pharmaceutically acceptable carrier or excipient.

[0012] In addition, the invention also provides the use of the pharmaceutical composition described in the above technical solution in the preparation of anti-liver fibrosis drugs. Also, the preparation method of the pharmaceutical composition is to first prepare compounds 1-8 according to the preparation method of the above compounds, and then take one of the compounds 1-8 or any combination thereof, and add a pharmaceutically acceptable carrier.

[0013] When at least one of the compounds 1-8 is used to prepare an anti-liver fibrosis drug, the present invention preferably uses the compounds 1-8 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-mentioned compounds 1-8 and a pharmaceutically acceptable carrier or excipient. In the present invention, the pharmaceutically acceptable carrier or excipient 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 or excipient, and any pharmaceutically acceptable carrier and / or excipient 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 1-8 can be directly mixed with a pharmaceutically acceptable carrier or excipient. 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 described in the above technical solution in the preparation of an anti-liver fibrosis drug. The present invention has no particular 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 fibrosis 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 1-8 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 can preferably be 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 trimer compounds, artemsieverolactones AH (1-8).

[0020] 2. The present invention provides a new method for preparing new compounds 1-8, which has readily available raw materials, is easy to operate, and is suitable for industrial production.

[0021] 3. The present invention provides a pharmaceutical composition with new compounds 1-8 as active ingredients, providing a new anti-liver fibrosis drug with better medicinal effects.

[0022] 4. Compounds 1-8 of the present invention have strong cytotoxic activity against hepatic stellate cells (HSC-LX2), and compounds 1, 2 and 4 have strong cytotoxic activity with IC 50 The values ​​were 49.2, 37.8 and 64.7 μM, respectively, and the activity was 2-4 times that of the positive drug silybin; compounds 3 and 7 also showed certain cytotoxic activity, and their IC50 The values ​​were 94.4 and 117.1 μM, respectively, which were better than the positive drug silybin. The other compounds had certain HSC-LX2 cytotoxic activity, and their inhibition rates were between 25.1-40.6% at a concentration of 200 μg / mL. Description of the drawings:

[0023] Figure 1 is the structural formula of compound 1-8 of the present invention, artemsieverolactones AH (1-8);

[0024] Figure 2 Schematic diagram of the X-ray single crystal diffraction structure of compound 1, artemsieverolactones A of the present invention. Specific implementation method:

[0025] In order to better understand the essence of the present invention, the test examples and embodiments of the present invention are used below in conjunction with the accompanying drawings to further illustrate the sesquiterpene trimer compound of the present invention, artemsieverolactones AH (1-8) and its preparation method, structural identification, and pharmacological effects, but the present invention is not limited to these test examples and embodiments.

[0026] 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 technical solutions obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] Embodiment 1:

[0028] Preparation of the sesquiterpenoid trimer compound of the present invention, artemsieverolactones AH (1-8):

[0029] The dried aerial part of Artemisia sieversiana (33 kg) was crushed and extracted twice with 95% ethanol for 4 days each time. The ethanol extracts were combined and the ethanol extract was recovered under reduced pressure. The extract was dispersed in water and extracted with ethyl acetate. The ethyl acetate extract was then concentrated to obtain the ethyl acetate extract. The ethyl acetate extract was then subjected to silica gel column chromatography with acetone-petroleum ether in a v / v ratio of 5:95, 10:90, 20:80, 30:7 0 and 100:0 as eluent, gradient elution gave five fractions Fr.A-Fr.E (149g, 101g, 98g, 153g and 164g); fraction Fr.D was eluted by silica gel column chromatography with methanol-chloroform at v / v 5:95, 10:90 and 20:80 to give five fractions Fr.D1-Fr.D5 (25g, 10g, 60g, 30g and 25g); fraction Fr.D4 (30g) was chromatographed on MCI gel CHP 20P column, gradient elution with water-methanol at 10:90 and 0:100 and semi-preparative HPLC to give compounds 1 (22mg), 2 (4mg), 3 (9mg), 4 (10mg), 5 (5mg), 6 (6mg), 7 (3mg), 8 (5mg).

[0030] Structural data of compounds 1-8:

[0031] Melting point X-4B micro melting point instrument (Shanghai Precision Scientific Instrument Co., Ltd.); specific rotation was measured by Autopol VI polarimeter (Rudolph Research Analytical, Hackettstown, USA); infrared spectrum was measured by ATR attenuated total reflection-diamond crystal (ATR ITX-DIAMOND) method, using NICOLET iS10 infrared spectrometer (Thermo Fisher Scientific, Madison, USA); UV spectrum was measured by UV-2401PC UV spectrometer (Shimadzu, Kyoto, Japan); ECD spectrum was measured by Applied Photophysics circular dichroism spectrometer (Applied Photophysics, Surrey, UK); NMR spectrum was measured by Avance III 600 (Bruker, Bremerhaven, Germany) superconducting nuclear magnetic resonance spectrometer was used for determination, and TMS (tetramethylsilane) was used as the internal standard. High-resolution mass spectrometry was determined using Shimadzu LC-MS-IT-TOF (Shimadzu, Kyoto, Japan), Agilent UPLC / Q-TOF and G6230 mass spectrometer (Agilent Technologies, Santa Clara, USA). Thin-layer chromatography silica gel plate HSGF254 was purchased from Yantai Jiangyou Silica Gel Development Co., Ltd.; column chromatography silica gel (200-300 mesh) was produced by Linyi Haixiang Chemical Co., Ltd.; column chromatography dextran gel LH-20 was purchased from GE Healthcare Bio-Sciences AB; high-performance liquid chromatograph was purchased from Shimadzu, and 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 was Agilent-Eclipse XDB-C18 (5 μm, 9.4×250 mm); chromatographically pure acetonitrile was purchased from Merida, and deionized channel water was purified by MingCheTM-D 24UV Merk Millipore system; medium pressure liquid phase (Dr Flash-Ⅱ) was a product of Shanghai Lisui Company, and MCI column was from Mitsubishi Corporation of Japan, model CHP-20P (75-150 μm); analytically pure methanol and acetonitrile were purchased from Tianjin Damao Chemical Reagent Factory; the color developer was 10% H 2 SO 4 -EtOH solution.

[0032]

[0033] Artemsieverolactone A(1)

[0034] Molecular formula: C 45 H 58 O 9

[0035] Molecular weight: 742

[0036] Appearance: Colorless needle crystal

[0037] Optical rotation: (c 0.071, methanol)

[0038] HRESIMS (–) m / z: Observed value 787.4063 [M+HCOO] – , calculated value 787.4063[M+HCOO] – .

[0039] IR max :3438,1758,1631,1454,1381,1241,1176,1055cm –1 .

[0040] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1.

[0041]

[0042] Artemsieverolactone B(2)

[0043] Molecular formula: C 45 H 58 O 9

[0044] Molecular weight: 742

[0045] Appearance: White powder

[0046] Optical rotation: (c 0.041, methanol)

[0047] HRESIMS (–) m / z: Observed value 787.4051 [M+HCOO] – , calculated value 787.4063[M+HCOO] – .

[0048] IR max :3439,1757,1631,1455,1381,1225,1178,1031cm -1 .

[0049] ECD(Methanol)λ max(Δε):201(+13.95),227(–1.16)nm.

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

[0051]

[0052] Artemsieverolactone C(3)

[0053] Molecular formula: C 45 H 58 O 10

[0054] Molecular weight: 758

[0055] Appearance: White powder

[0056] Optical rotation: (c 0.10, methanol)

[0057] HRESIMS (–) m / z: Observed value 803.4017 [M+HCOO] – , calculated value 803.4012 [M+HCOO] – .

[0058] IR max :3438,1760,1632,1454,1297,1178,1003cm -1 .

[0059] ECD(Methanol)λ max (Δε):213(+19.22)nm.

[0060] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1.

[0061]

[0062] Artemsieverolactone D(4)

[0063] Molecular formula: C 45 H 58 O 9

[0064] Molecular weight: 742

[0065] Appearance: White powder

[0066] Optical rotation: (c 0.065, methanol)

[0067] HRESIMS (–) m / z: Observed value 787.4063 [M+HCOO] – , calculated value 787.4063[M+HCOO] – .

[0068] IR max :3436,1761,1633,1454,1382,1318,1178,1148,1000cm -1 .

[0069] ECD(Methanol)λ max (Δε):195(+36.29),225(–5.97)nm.

[0070] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1.

[0071]

[0072] Artemsieverolactone E(5)

[0073] Molecular formula: C 45 H 58 O 10

[0074] Molecular weight: 758

[0075] Appearance: White powder

[0076] Optical rotation: (c 0.051, methanol)

[0077] HRESIMS (–) m / z: Observed value 803.4016 [M+HCOO] – , calculated value 803.4012 [M+HCOO] – .

[0078] IR max :3437,1764,1632,1455,1381,1319,1240,1150,993cm -1 .

[0079] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 2.

[0080]

[0081] Artemsieverolactone F(6)

[0082] Molecular formula: C 45 H 58 O 9

[0083] Molecular weight: 742

[0084] Appearance: White powder

[0085] Optical rotation: (c 0.069, methanol)

[0086] HRESIMS (–) m / z: Observed value 787.4053 [M+HCOO] – , calculated value 787.4063[M+HCOO] – .

[0087] IR max :3436,1767,1632,1455,1382,1315,1178,1141cm –1 .

[0088] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 2.

[0089]

[0090] Artemsieverolactone G(7)

[0091] Molecular formula: C 45 H 58 O 9

[0092] Molecular weight: 742

[0093] Appearance: White powder

[0094] Optical rotation: (c 0.073, methanol)

[0095] HRESIMS (–) m / z: Observed value 787.4052 [M+HCOO] – , calculated value 787.4063[M+HCOO] – .

[0096] IR max :3438,1760,1635,1457,1382,1319,1176cm -1 .

[0097] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 2.

[0098]

[0099] Artemsieverolactone H(8)

[0100] Molecular formula: C 45 H 58 O 9

[0101] Molecular weight: 742

[0102] Appearance: White powder

[0103] Optical rotation: (c 0.053, methanol)

[0104] HRESIMS (–) m / z: Observed value 787.4063 [M+HCOO] – , calculated value 787.4063[M+HCOO] – .

[0105] IR max :3462,1755,1638,1457,1382,1176,1117cm -1 .

[0106] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 2.

[0107] Table 1. Compounds 1-4 1 H and 13 C-NMR data (600 MHz, δ in ppm, J in Hz, CDCl 3 )

[0108]

[0109] "ol" means the signals overlap and the coupling constant cannot be calculated

[0110] Table 2. Compounds 5-8 1 H and 13 C-NMR data (600 MHz, δ in ppm, J in Hz, CDCl 3 )

[0111]

[0112] "ol" means the signals overlap and the coupling constant cannot be calculated

[0113] Embodiment 2:

[0114] Cytotoxic activity of artemsieverolactones AH(1-8), a sesquiterpenoid trimer, against HSC-LX2 cells.

[0115] 1. Materials and Methods

[0116] 1.1 Materials

[0117] Human hepatic stellate cell line LX2 (HSC-LX2) was purchased from Shanghai Jining Biotechnology Co., Ltd.; RPMI-1640 culture medium and fetal bovine serum were purchased from Gibco BRL (NY, USA); MTT was purchased from Guangzhou Saiguo Biotechnology Co., Ltd.

[0118] 1.2 Instruments

[0119] Flex Station 3 multifunctional microplate reader (Bio-RAD 680, USA); analytical balance (AG135, Metler Toledo, China); constant temperature box (DHP-9082, Shanghai).

[0120] 1.3 Experimental process

[0121] The cytotoxic activity of the samples on HSC-LX2 cells was determined by MTT method. HSC-LX2 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum. Cells in logarithmic phase were taken and plated at 1×10 4 / well were inoculated in a 96-well plate. After 24 hours, the maintenance solution was replaced with culture medium containing different concentrations of the test sample. At the same time, a cell control group with only maintenance solution was set up, and silybin was used as a positive drug control. After 48 hours of culture, the culture medium was discarded and 100μL MTT solution (concentration of 1mg / mL) was added; incubated in a 37℃ incubator for 4 hours, the MTT solution was discarded and 100μL DMSO was added to dissolve the formazan crystals; finally, the absorbance value of each well was detected at 490nm using an enzyme reader. The inhibition rate of HSC-LX2 cells was calculated as inhibition rate (%) = [A(blank) – A(sample)] / A(blank) × 100%. Its half inhibitory concentration (50% inhibitory concentration, IC 50 ) was calculated using Graphpad Prism 5 software.

[0122] 2. Results

[0123] All the isolated compounds were evaluated for their in vitro HSC-LX2 cytotoxic activity (Table 3). Compounds 1-8 all had strong cytotoxic activity against hepatic stellate cells (HSC-LX2), and compounds 1, 2 and 4 had strong cytotoxic activity with IC 50 The values ​​were 49.2, 37.8 and 64.7 μM, respectively, and the activity was 2-4 times that of the positive drug silybin; compounds 3 and 7 also showed certain cytotoxic activity, and their IC 50 The values ​​were 94.4 and 117.1 μM, respectively, which were better than the positive drug silybin. The other compounds had certain HSC-LX2 cytotoxic activity, and their inhibition rates were between 25.1-40.6% at a concentration of 200 μg / mL.

[0124] Table 3. HSC-LX2 cytotoxic activity of compounds

[0125]

[0126]

[0127] Inhibition rate and IC 50 All were obtained from three independent experiments.

[0128] 3. Conclusion

[0129] The experimental results showed that compounds 1-8 had strong cytotoxic activity against hepatic stellate cells (HSC-LX2), and compounds 1, 2 and 4 had strong cytotoxic activity with IC 50 The values ​​were 49.2, 37.8 and 64.7 μM, respectively, and the activity was 2-4 times that of the positive drug silybin; compounds 3 and 7 also showed certain cytotoxic activity, and their IC 50 The values ​​were 94.4 and 117.1 μM, respectively, which were better than the positive drug silybin. The other compounds had certain HSC-LX2 cytotoxic activity, and their inhibition rates were between 25.1-40.6% at a concentration of 200 μg / mL.

[0130] The above results indicate that artemsieverolactones AH(1-8) in Artemisia annua can be used as drugs for the preparation of drugs against liver fibrosis.

[0131] Formulation Examples 1-7:

[0132] 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 1-8 described in the present invention can be prepared into different formulations, and no specific restrictions are imposed on the specific reagents and operations.

[0133] 1. Dissolve at least one of compounds 1-8 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.

[0134] 2. Dissolve at least one of the compounds 1-8 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, dispense it into ampoules, freeze-dry it at low temperature, and seal it with sterile sealing to obtain a powder injection.

[0135] 3. Add at least one of compounds 1-8 to an excipient at a mass ratio of 9:1 to the excipient to prepare a powder.

[0136] 4. Add at least one of compounds 1-8 to the excipient at a mass ratio of 5:1, and granulate and tablet.

[0137] 5. Prepare at least one of compounds 1-8 into an oral liquid according to a conventional oral liquid preparation method.

[0138] 6. Add at least one of compounds 1-8 to the excipient at a mass ratio of 5:1 to the excipient to prepare capsules.

[0139] 7. Add at least one of compounds 1-8 to an excipient at a mass ratio of 5:1 to the excipient to prepare granules.

[0140] As can be seen from the above examples, the present invention provides a new sesquiterpenoid compound in Artemisia grandis and its preparation method and application, a pharmaceutical composition and its application. The eight new sesquiterpenoid compounds provided by the present invention have different degrees of cytotoxic activity against human hepatic stellate cells HSC-LX2, can be combined with a pharmaceutically acceptable carrier or excipient to form a pharmaceutical composition, and can be used to prepare an anti-liver fibrosis drug.

[0141] 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. A sesquiterpene trimer compound 1-8 as shown in the following structural formula (I), 2. The method for preparing the sesquiterpene trimer compound 1-8 represented by the structural formula (I) according to claim 1, characterized in that: The method comprises the following steps: the dried aerial part of Artemisia grandis is crushed, and cold-extracted twice with 95% ethanol, each time for 4 days, the ethanol extract is combined, the ethanol extract is recovered under reduced pressure, the extract is dispersed in water and extracted with ethyl acetate, and then concentrated to obtain the ethyl acetate extraction part, and then the ethyl acetate extraction part is subjected to silica gel column chromatography, acetone-petroleum ether is used as eluent in a ratio of 5:95, 10:90, 20:80, 30:70 and 100:0 by v / v, and gradient elution is performed to obtain five fractions Fr.A-Fr.E; the fraction Fr.D is subjected to silica gel column chromatography, and methanol-chloroform is used in a ratio of 5:95, 10:90 and 20:80 by v / v to obtain five fractions Fr.D1-Fr.D5; the fraction Fr.D4 is subjected to MCI gel CHP 20P column chromatography, and water-methanol is used as gradient elution with a ratio of 10:90 and 0:100 and semi-preparative HPLC to prepare compounds 1, 2, 3, 4, 5, 6, 7 and 8.

3. Use of the sesquiterpene trimer compounds 1-8 represented by the structural formula (I) according to claim 1 in the preparation of anti-liver fibrosis drugs.

4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises at least one of the compounds 1-8 represented by the structural formula (I) according to claim 1 and a pharmaceutically acceptable carrier.

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

6. The method for preparing the pharmaceutical composition according to claim 4, characterized in that: The method comprises the following steps: the dried aerial part of Artemisia grandis is crushed, and cold-extracted with 95% ethanol twice, each time for 4 days, the ethanol extract is combined, the ethanol extract is recovered under reduced pressure, the extract is dispersed in water and extracted with ethyl acetate, and then concentrated to obtain the ethyl acetate extraction part, and then the ethyl acetate extraction part is subjected to silica gel column chromatography, acetone-petroleum ether is used as an eluent at a ratio of 5:95, 10:90, 20:80, 30:70 and 100:0 in v / v, and gradient elution is performed to obtain five fractions Fr.A-Fr.E; the fraction Fr.D is subjected to silica gel column chromatography and eluted with methanol-chloroform at a ratio of 5:95, 10:90 and 20:80 in v / v to obtain five fractions Fr.D1-Fr.D5; the fraction Fr.D4 is subjected to MCI gel chromatography Compounds 1, 2, 3, 4, 5, 6, 7, and 8 were prepared by CHP20P column chromatography, gradient elution with water-methanol 10:90 and 0:100, and semi-preparative HPLC; then one of compounds 1-8 or any combination thereof was added with a pharmaceutically acceptable carrier.

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