Sesquiterpene dimer in artemisia selengensis, pharmaceutical composition of sesquiterpene dimer, preparation method of sesquiterpene dimer and application of sesquiterpene dimer

Abstract

The invention provides a sesquiterpene dimer in artemisia selengensis, a pharmaceutical composition of the sesquiterpene dimer and a preparation method and application of the sesquiterpene dimer, and belongs to the technical field of medicines. According to the present invention, the eight new sesquiterpene dimers represented by the structural formula of the present invention, artemselenoids A-H (1-8) have significant inhibitory activity on three liver cancer cells (HepG2, Huh7 and SK-Hep-1), can form the pharmaceutical composition with the pharmaceutically acceptable carrier, and can be used for preparing the anti-liver cancer drug.

Description

Technical Field:

[0001] The present invention belongs to the field of pharmaceutical technology. Specifically, it relates to sesquiterpene dimer compounds artemselenoids A–H (1–8), their preparation methods, pharmaceutical compositions and their applications. Background Art:

[0002] Liver cancer is a common malignant tumor globally. Hepatocellular carcinoma (HCC) in primary liver cancer accounts for approximately 90%, which is highly malignant, has a poor prognosis, is prone to metastasis and recurrence, and the effects of traditional treatment methods are limited. Global cancer data in 2022 shows that the number of newly diagnosed liver cancer cases exceeded 865,000, ranking 6th in terms of incidence among cancers and 3rd in terms of the number of deaths. In China, the number of newly diagnosed liver cancer cases reached 4.1 million, accounting for 45.3% of the global total. The main therapeutic drugs for hepatocellular carcinoma are 5 kinase inhibitors (sorafenib, lenvatinib, regorafenib, cabozantinib, donafenib) and 3 monoclonal antibodies (nivolumab, pembrolizumab, ramucirumab). Although these drugs have greatly improved the survival status of HCC patients, they have the disadvantages of similar structures and overlapping targets, resulting in patients being prone to drug resistance. Therefore, the development of anti-liver cancer drugs with new mechanisms of action and high effectiveness is still an urgent need in China and even the world. Natural products have diverse chemical structures and significant biological activities, and are an important source for the development of anti-tumor drugs. 62% of the anti-cancer drugs approved by the FDA (1981 - 2019) are derived from natural products or inspired by natural products, which indicates the key role of natural products in the research and development of new anti-tumor drugs. The present invention aims to screen for new anti-liver cancer active ingredients from natural products based on a screening model of three liver cancer cell lines (HepG2, Huh7, and SK-Hep-1).

[0003] Artemisia selengensis Turcz. ex Besser is a perennial herbaceous plant of the genus Artemisia in the Compositae family, which is widely distributed in East Asia and grows mostly in wetland environments such as lakesides, riversides, and swamps. As a plant with both medicinal and edible uses, Artemisia selengensis has hemostatic, anti-inflammatory, and antitussive effects. Modern research shows that its extracts also have activities such as anti-hyperuricemia and antioxidant effects. Phytochemical studies have elucidated 71 chemical components of it, including polyynes, sesquiterpenes, sesquiterpene dimers, flavonoids, caffeoylquinic acid compounds, and other types of compounds.

[0004] So far, there is no report on the sesquiterpene dimer artemselenoids A–H (compounds 1–8) in the prior art, nor is there a report on compounds 1 - 8 and their pharmaceutical compositions as anti-liver cancer drugs. Summary of the Invention:

[0005] The object of the present invention is to provide a new class of sesquiterpene dimers with medicinal value, artemselenoids A–H (1–8), and their preparation methods and applications, a pharmaceutical composition containing artemselenoids A–H and their applications. These compounds have obvious inhibitory activity on the proliferation of three hepatocellular carcinoma cell lines (HepG2, Huh7, and SK-Hep-1), and can be used to prepare anti-hepatocellular carcinoma drugs.

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

[0007] The present invention provides a series of sesquiterpene dimer compounds, artemselenoids A–H (1–8), having the structure shown in the following formula (I):

[0008]

[0009] The present invention provides a preparation method of the above sesquiterpene dimer compounds 1-8. The dried aerial parts of Artemisia selengensis Turcz. are crushed and extracted twice with 90% ethanol by cold maceration for 4 days each time. The extraction solutions are combined and concentrated under reduced pressure to obtain an ethanol extract. It is dispersed in water and then extracted with ethyl acetate, and then concentrated to obtain an ethyl acetate extraction fraction. The ethyl acetate fraction is subjected to silica gel column chromatography and eluted with a gradient of acetone-petroleum ether mixed solvents (0:100, 1:200, 1:100, 2:98, 5:95, 10:90, 20:80, 40:60, 50:50, v / v) to obtain 6 fractions (Fr.1-Fr.6). Fraction Fr.4 is subjected to silica gel column chromatography and eluted with a gradient of acetone-petroleum ether mixed solvents (5:95, 10:90, 20:80, 40:60, 50:50, v / v) to obtain 6 fractions (Fr.4.1-Fr.4.6). Among them, fraction Fr.4.4 is subjected to repeated MCI gel CHP P20 column chromatography, silica gel column chromatography, gel column chromatography, reverse-phase column chromatography, and semi-preparative HPLC to obtain compounds 1 (3.5 mg), 2 (3.0 mg), 3 (2.0 mg), 4 (5.0 mg), 5 (4.0 mg), 6 (3.4 mg), 7 (8.0 mg), 8 (11.0 mg); then one or any combination of compounds 1–8 is taken, and a pharmaceutically acceptable carrier is added.

[0010] The present invention provides the application of the sesquiterpene dimer compounds 1-8 described in the above technical solutions in the preparation of anti-hepatocellular carcinoma drugs. The present invention has no special limitation on the method of the application, and a method well-known in the art can be selected.

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

[0012] In addition, the present invention also provides the use of the pharmaceutical composition described in the above technical solution in the preparation of anti-hepatocellular carcinoma drugs. Moreover, for the preparation method of the pharmaceutical composition, compounds 1-8 are first prepared according to the preparation method of the above compounds, and then one or any combination of compounds 1-8 is taken, and a pharmaceutically acceptable carrier is added.

[0013] When at least one of the compounds 1-8 is used for the preparation of anti-hepatocellular carcinoma drugs, 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 compounds 1-8 and a pharmaceutically acceptable carrier. In the present invention, the pharmaceutically acceptable carrier is preferably a solid, semi-solid or liquid diluent, filler and pharmaceutical product adjuvant. The present invention has no special limitation on the pharmaceutically acceptable carrier, and a pharmaceutically acceptable carrier well-known in the art, which is non-toxic and inert to humans and animals, can be selected.

[0015] The present invention has no special limitation on the preparation method of the pharmaceutical composition, and at least one of the compounds 1-8 can be directly mixed with a pharmaceutically acceptable carrier. The present invention has no special limitation on the mixing process, and 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 anti-hepatocellular carcinoma drugs. The present invention has no special limitation on the application method, and a method well-known in the art can be selected.

[0017] In the present invention, when the pharmaceutical composition is used for the preparation of anti-hepatocellular carcinoma drugs, 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 is preferably administered by 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 new sesquiterpene dimer compounds, artemselenoids A–H (1-8).

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

[0021] 3. The present invention provides a pharmaceutical composition comprising new compounds 1-8 as active ingredients, offering a new drug with good medicinal effects for treating liver cancer.

[0022] 4. Compounds 1-8 of the present invention exhibit inhibitory activity against three liver cancer cell lines. For HepG2 cells, compounds 3 and 8 showed strong inhibitory activity, with IC 50 values of 7.4 and 5.1 μM respectively, superior to the positive control drug sorafenib (IC 50 , 9.9 μM). Compounds 1, 4, 5 and 6 showed moderate activity, with IC 50 values in the range of 17.1 - 33.9 μM, while compounds 2 and 7 showed weak activity, with IC 50 values of 65.5 and 66.2 μM respectively; for Huh7 cells, compounds 3 and 8 showed significant inhibitory activity, with IC 50 values of 5.9 and 8.6 μM respectively, close to the positive drug sorafenib (IC 50 , 5.4 μM). Compounds 1, 4 and 5 showed moderate inhibitory activity, with IC 50 values in the range of 22.9 - 29.5 μM, while compounds 2, 6 and 7 showed slightly weaker inhibitory activity, with IC 50 values in the range of 50.7 - 70.4 μM; for SK-Hep-1 cells, compound 8 showed significant inhibitory activity, with IC 50 of 9.6 μM, superior to the positive drug sorafenib (IC 50 , 13.1 μM). The IC 50 values of compounds 1-7 were in the range of 20.9 - 65.4 μM. BRIEF DESCRIPTION OF THE DRAWINGS:

[0023] Figure 1 It is a schematic diagram of the structural formula of compounds 1-8 of the present invention. DETAILED DESCRIPTION OF THE INVENTION:

[0024] To better understand the essence of the present invention, the following will further illustrate the sesquiterpene dimer compounds, artemselenoids A-H (1-8) of the present invention, their preparation methods, structural identification, and pharmacological effects with reference to the drawings, test examples and examples of the present invention, but the present invention is not limited by these test examples and examples.

[0025] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0026] Example 1:

[0027] Preparation of the sesquiterpene dimer compounds, artemselenoids A–H (1–8) of the present invention:

[0028] The dried aerial parts of Artemisia sieversiana (22 kg) were crushed and extracted twice with 90% ethanol by cold maceration for 4 days each time. The extraction solutions were combined and concentrated under reduced pressure to obtain an ethanol extract. It was dispersed in water and then extracted with ethyl acetate, and subsequently concentrated to obtain an ethyl acetate extract fraction. The ethyl acetate fraction was subjected to silica gel column chromatography and eluted with a gradient of acetone - petroleum ether mixed solvents (0:100, 1:200, 1:100, 2:98, 5:95, 10:90, 20:80, 40:60, 50:50, v / v) to obtain 6 fractions (Fr.1-Fr.6). Fraction Fr.4 was subjected to silica gel column chromatography and eluted with a gradient of acetone - petroleum ether mixed solvents (5:95, 10:90, 20:80, 40:60, 50:50, v / v) to obtain 6 fractions (Fr.4.1-Fr.4.6). Among them, fraction Fr.4.4 was subjected to repeated MCI gel CHP P20 column chromatography, silica gel column chromatography, gel column chromatography, reverse-phase column chromatography and semi-preparative HPLC to obtain compounds 1 (3.5 mg), 2 (3.0 mg), 3 (2.0 mg), 4 (5.0 mg), 5 (4.0 mg), 6 (3.4 mg), 7 (8.0 mg), 8 (11.0 mg).

[0029] Structural data of compounds 1–8:

[0030] The specific rotation was measured by an Autopol VI polarimeter (Rudolph Research Analytical, Hackettstown, USA); the infrared spectrum was determined by the ATR attenuated total reflection-diamond crystal (ATR ITX-DIAMOND) method using a NICOLET iS10 Fourier transform infrared spectrometer (Thermo Fisher Scientific, Madison, USA); the ultraviolet spectrum was measured with a UV-2401PC ultraviolet spectrometer (Shimadzu, Kyoto, Japan); the ECD spectrum was recorded on an Applied Photophysics circular dichroism spectrometer (Applied Photophysics, Surrey, UK); the nuclear magnetic resonance spectra were obtained using an Avance III 600 (Bruker, Bremerhaven, Germany) superconducting nuclear magnetic resonance spectrometer; high-resolution mass spectra were measured on a Shimadzu LCMS-IT-TOF (Shimadzu, Kyoto, Japan), an Agilent UPLC / Q-TOF and a G6230 mass spectrometer (Agilent Technologies, Santa Clara, USA); the TLC silica gel plate HSGF254 was purchased from Yantai Jiangyou Silica Gel Development Co., Ltd.; the column chromatography silica gel (200 - 300 mesh, 300 - 400 mesh) was produced by Linyi Haixiang Chemical Co., Ltd.; the column chromatography Sephadex LH-20 was purchased from GE Healthcare Bio-Sciences AB; the high-performance liquid chromatography instrument was purchased from Shimadzu Corporation, with the controller model CBM-20A, the pump model LC-20AR, the detector model SPD-M20A, the column oven model AT-350, and the chromatographic column models Agilent-Eclipse XDB-C18 (5 μm, 9.4 × 250 mm) and YMC-Pack ODA-A (5 μm, 10.0 × 250 mm); the chromatographic grade acetonitrile and methanol were purchased from Merck Millipore, and the deionized water was purified by a MingCheTM-D 24UV Merk Millipore system; the medium-pressure liquid chromatography (Dr Flash-II) was a product of Shanghai Lishui Co., Ltd., and the MCI column was from Mitsubishi Corporation of Japan, model CHP-20P (75 - 150 μm); RP-C 18 was purchased from Fuji Chemical Industry Co., Ltd., with a specification of 40 - 75 μm, and the developer was 10% H 2 SO 4 -EtOH solution.

[0031]

[0032] artemselenoid A(1)

[0033] Molecular formula: C 30 H 34 O 6

[0034] Molecular weight: 490

[0035] Appearance: White amorphous powder

[0036] Optical rotation: (c 0.11, methanol)

[0037] HRESIMS(+) m / z: Experimental value 491.2426 [M+H] + , Calculated value 491.2428 [M+H] + .

[0038] IR v max : 3437, 1765, 1665, 1636, 1619, 1221, 1149 cm –1 .

[0039] ECD (methanol) λ max (Δε) 196 (+24.55), 221 (-16.85), 252 (+8.06) nm.

[0040] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1 and Table 3.

[0041]

[0042] artemselenoid B(2)

[0043] Molecular formula: C 32 H 40 O 8

[0044] Molecular weight: 552

[0045] Appearance: White amorphous powder

[0046] Optical rotation: (c 0.13, methanol)

[0047] HRESIMS(+) m / z: Experimental value 553.2791 [M+H] + , Calculated value 553.2796 [M+H] – . IR v max : 3436, 1760, 1630, 1458, 1382, 1149, 986 cm -1 .

[0048] ECD (methanol) λ max (Δε) 209 (-6.39), 235 (-6.08), 3.20 (-1.15) nm.

[0049] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1 and Table 3.

[0050]

[0051] artemselenoid C(3)

[0052] Molecular formula: C 30 H 36 O 6

[0053] Molecular weight: 492

[0054] Appearance: White amorphous powder

[0055] Optical rotation: (c 0.07, methanol)

[0056] HRESIMS(+) m / z: Experimental value 493.2580 [M+H] + , Calculated value 493.2585 [M+H] + . IR v max : 3460, 1749, 1637, 1459, 1384, 1139 cm -1 .

[0057] ECD (methanol) λ max (Δε) 195 (-10.65), 204 (+0.68), 224 (-6.93) nm.

[0058] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 1 and Table 3.

[0059]

[0060] artemselenoid D(4)

[0061] Molecular formula: C 30 H 36 O 6

[0062] Molecular weight: 492

[0063] Appearance: White amorphous powder

[0064] Optical rotation: (c 0.04, methanol)

[0065] HRESIMS(+) m / z: experimental value 493.2594 [M+H] + , calculated value 493.2585 [M+H] + . IR ν max : 3439, 1748, 1638, 1458, 1384, 1126 cm -1 .

[0066] ECD (methanol) λ max (Δε) 195 (-8.13), 206 (-3.37), 223 (-5.49) nm.

[0067] 1 1H NMR and 13 13C NMR (DEPT) data are shown in Table 1 and Table 3.

[0068]

[0069] artemselenoid E(5)

[0070] Molecular formula: C 31 H 38 O 6

[0071] Molecular weight: 506

[0072] Appearance: white amorphous powder

[0073] Optical rotation: (c 0.09, methanol)

[0074] HRESIMS(+) m / z: experimental value 525.2472 [M+Na] + , calculated value 525.2483 [M+Na] + . IR ν max : 3465, 1763, 1637, 1458, 1383, 1129 cm -1 .

[0075] ECD (methanol) λ max 199 (+15.70), 224 (-1.57) nm.

[0076] 1 1H NMR and 13 13C NMR (DEPT) data are shown in Table 2 and Table 3.

[0077]

[0078] artemselenoid F(6)

[0079] Molecular formula: C 30 H 36 O 8

[0080] Molecular weight: 524

[0081] Appearance: White amorphous powder

[0082] Optical rotation: (c 0.05, methanol)

[0083] HRESIMS(+) m / z: Experimental value 525.2472 [M+H] + , Calculated value 525.2483 [M+H] + . IR ν max : 3450, 1754, 1632, 1456, 1384, 1150 cm –1 .

[0084] ECD (methanol) λ max 197 (+13.41), 213 (+15.29), 238 (-15.31) nm.

[0085] 1 H NMR and 13 C NMR (DEPT) data are shown in Table 2 and Table 3.

[0086]

[0087] artemselenoid G(7)

[0088] Molecular formula: C 30 H 36 O 8

[0089] Molecular weight: 524

[0090] Appearance: White amorphous powder

[0091] Optical rotation: (c 0.08, methanol)

[0092] HRESIMS(+) m / z: Experimental value 525.2480 [M+H] + , Calculated value 525.2483 [M+H] + . IR ν max : 3465, 1651, 1460, 1382, 1175, 986 cm -1。

[0093] ECD (methanol) λ max (Δε) 198 (-21.33), 229 (+21.34), 280 (+10.47), 295 (+11.18) nm. 1 HNMR and 13 C NMR (DEPT) data are shown in Tables 2 and 3.

[0094]

[0095] artemselenoid H(8)

[0096] Molecular formula: C 30 H 34 O 6

[0097] Molecular weight: 490

[0098] Appearance: white amorphous powder

[0099] Optical rotation: (c 0.13, methanol)

[0100] HRESIMS(+) m / z: experimental value 491.2423 [M+H] + , calculated value 491.2428 [M+H] + . IR v max : 3438, 1756, 1683, 1637, 1618, 1444, 1258, 1150, 1007 cm -1 .

[0101] ECD (methanol) λ max (Δε) 197 (+18.09), 225 (+0.25), 250 (+6.02) nm.

[0102] 1 H NMR and 13 C NMR (DEPT) data are shown in Tables 2 and 3.

[0103] Table 1. 1 H NMR (600 MHz) data (δ in ppm, J in Hz, CDCl 3 )

[0104]

[0105] a“ol” is used to indicate overlapped signals, for which the coupling constants could not be read.

[0106] Table 2. 1 1H NMR (600 MHz) data (δ in ppm, J in Hz, CDCl 3 )

[0107]

[0108] a “ol” is used to indicate overlapped signals, for which the coupling constants could not be read.

[0109] Table 3. 13 13C NMR data (150 MHz, δ in ppm, J in Hz, CDCl 3 )

[0110]

[0111] Example 2:

[0112] The inhibitory activities of sesquiterpene dimer compounds, artemselenoids A–H (1–8) on the proliferation of three hepatoma cell lines (HepG2, Huh7 and SK-Hep-1).

[0113] 1. Materials and methods

[0114] 1.1 Materials

[0115] Human hepatoma cell lines (HepG2, Huh7 and SK-Hep-1) were purchased from Shanghai Jining Biotechnology Co., Ltd.; RPMI-1640 medium and fetal bovine serum were both purchased from Gibco BRL (NY, USA); MTT was purchased from Guangzhou Saiguo Biotechnology Co., Ltd.

[0116] 1.2 Instruments

[0117] Flex Station 3 desktop multi-functional microplate reader (Bio-RAD 680, USA); analytical balance (AG135, Metler Toledo, China); incubator (DHP-9082, Shanghai).

[0118] 1.3 Experimental procedures

[0119] The MTT method was used to evaluate the inhibitory activity of the test samples against HepG2, Huh7, and SK-Hep-1 cells. HepG2 and SK-Hep-1 cells were cultured in MEM medium with 10% fetal bovine serum in an incubator at 37 °C and 5% CO 2 , while Huh7 cells were cultured in Dulbecco's modified Eagle medium with 10% fetal bovine serum. Cells (5×10 3 cells / well) were seeded into 96-well plates and cultured at 37 °C and 5% CO 2 for 24 hours. Then, the medium containing different concentrations of the test samples was added, and a blank control group was set up, with sorafenib as the positive control. After incubation for 48 h, the culture medium was aspirated, and then 100 μL of MTT solution (concentration 1 mg / mL) was added to each well; after culturing for 4 h, the MTT solution was aspirated and 100 μL of DMSO was added; the absorbance value was measured at 490 nm using a multi-functional microplate reader. The formula: Inhibition rate (%) = [A (对照) – A (实验) / A (对照) × 100% was used to calculate the inhibition rate. Graphpad Prism 5 software was used to calculate the IC 50 .

[0120] 2. Results

[0121] The results of the inhibitory activities of compounds 1–8 against three hepatocarcinoma cell lines (HepG2, Huh7, and SK-Hep-1) are shown in Table 4. Compounds 3, 4, and 5 showed strong inhibitory activities, with IC 50 values of 52.6, 58.4, and 54.8 μM, respectively, which were 3 times that of the positive drug silybin (IC 50 , 149.4 μM); the IC 50 value of compound 2 was 2 times that of the positive drug, being 105.2 μM; the IC 50 value of compound 1 was comparable to that of the positive drug, being 143.3 μM. In comparison, the activity of compound 6 was weak, with an IC 50 value of 194.6 μM, while the inhibition rate of compound 7 was 39.0% at a concentration of 200 μM.

[0122] Table 4. Proliferation inhibitory activities of compounds 1-8 against three hepatocarcinoma cell lines

[0123]

[0124]

[0125] a IC 50 was obtained from three independent experiments; bSorafenib was used as the positive control.

[0126] 3. Conclusions

[0127] The experimental results showed that compounds 1–8 had varying degrees of inhibitory activity against three hepatocellular carcinoma cell lines (HepG2, Huh7, and SK-Hep-1). For HepG2 cells, compounds 3 and 8 showed strong inhibitory activity, with IC 50 values of 7.4 and 5.1 μM, respectively, which were superior to the positive control drug sorafenib (IC 50 , 9.9 μM). Compounds 1, 4, 5, and 6 showed moderate activity, with IC 50 values in the range of 17.1–33.9 μM, while compounds 2 and 7 showed weak activity, with IC 50 values of 65.5 and 66.2 μM, respectively. For Huh7 cells, compounds 3 and 8 showed significant inhibitory activity, with IC 50 values of 5.9 and 8.6 μM, respectively, which were close to the positive drug sorafenib (IC 50 , 5.4 μM). Compounds 1, 4, and 5 showed moderate inhibitory activity, with IC 50 values in the range of 22.9–29.5 μM, while compounds 2, 6, and 7 showed slightly weaker inhibitory activity, with IC 50 values in the range of 50.7–70.4 μM. For SK-Hep-1 cells, compound 8 showed significant inhibitory activity, with an IC 50 value of 9.6 μM, which was superior to the positive drug sorafenib (IC 50 , 13.1 μM). The IC 50 values of compounds 1–7 were in the range of 20.9–65.4 μM.

[0128] The above results indicate that the sesquiterpene dimer compounds, artemselenoids A–H (1–8) from Artemisia selengensis Turcz. ex Bess., can be used as drugs for the preparation of anti-hepatocellular carcinoma drugs.

[0129] Formulation Examples 1-8:

[0130] In the following formulation examples, conventional reagents were selected and formulation preparation was carried out according to existing conventional methods. This application example only demonstrates that at least one of the compounds 1-8 of the present invention can be prepared into different formulations, and specific reagents and operations are not specifically limited:

[0131] 1. At least one of compounds 1-8 was dissolved in DMSO, and then water for injection was added according to the conventional method, followed by fine filtration, filling, and sterilization to prepare an injection solution, and the concentration of the injection solution was 0.5-5 mg / mL.

[0132] 2. Dissolve at least one of Compounds 1-8 in DMSO, then dissolve it in sterile injection water, stir to dissolve, filter with a sterile suction funnel, and then filter through sterile fine filtration. Aliquot into ampoules, freeze-dry at low temperature and then seal aseptically to obtain a powder injection.

[0133] 3. Add at least one of Compounds 1-8 to an excipient in a mass ratio of 9:1 to the excipient to prepare a powder.

[0134] 4. Add at least one of Compounds 1-8 to an excipient in a mass ratio of 5:1 to the excipient, granulate and press into tablets.

[0135] 5. Prepare an oral liquid by a conventional method for preparing oral liquids using at least one of Compounds 1-8.

[0136] 6. Add at least one of Compounds 1-8 to an excipient in a mass ratio of 5:1 to the excipient to prepare capsules.

[0137] 7. Add at least one of Compounds 1-8 to an excipient in a mass ratio of 5:1 to the excipient to prepare granules.

[0138] As can be seen from the above examples, the present invention provides a compound in Artemisia selengensis Turcz., its preparation method and application, a pharmaceutical composition and its application. The eight new sesquiterpene compounds provided by the present invention have different degrees of inhibitory activity against the proliferation of three hepatoma cell lines (HepG2, Huh7 and SK-Hep-1), can form a pharmaceutical composition with a pharmaceutically acceptable carrier or excipient, and can be used for the preparation of anti-hepatoma drugs.

[0139] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. Sesquiterpene dimer compounds 1-8 as shown in the following structural formula, 2. The method for preparing the sesquiterpene dimer compound 1-8 represented by the structural formula according to claim 1, characterized in that: The method comprises the following steps: the dried aerial part of Artemisia selengensis is crushed, and the aerial part is extracted twice by cold soaking with 90% ethanol, each time for 4 days, and the extracts are combined and concentrated under reduced pressure to obtain an ethanol extract; the aerial part is dispersed with water, extracted with ethyl acetate, and then concentrated to obtain an ethyl acetate extraction part; the ethyl acetate extraction part is subjected to silica gel column chromatography, acetone-petroleum ether is calculated by v / v, 0:100, 1:200, 1:100, 2:98, 5:95, 10:90, 20:80, 40:60, 50:50 as eluent, and gradient elution is performed to obtain 6 fractions Fr.1-Fr.6; the fraction Fr.4 is subjected to silica gel column chromatography, acetone-petroleum ether is calculated by v / v, 5:95, 10:90, 20:80, 40:60, 50:50 as eluent, and gradient elution is performed to obtain 6 fractions Fr.4.1-Fr.4.6, wherein the fraction Fr.4.4 is subjected to repeated MCI gel chromatography Compounds 1, 2, 3, 4, 5, 6, 7, 8 were obtained by CHP P20 column chromatography, silica gel column chromatography, gel column chromatography, reverse phase column chromatography and semi-preparative HPLC.

3. Use of the sesquiterpene dimer compounds 1-8 represented by the structural formula according to claim 1 in the preparation of anti-liver cancer 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 of 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: the dried aerial part of Artemisia selengensis is crushed, and the aerial part is extracted twice by cold soaking with 90% ethanol, each time for 4 days, and the extracts are combined and concentrated under reduced pressure to obtain an ethanol extract; the aerial part is dispersed with water, extracted with ethyl acetate, and then concentrated to obtain an ethyl acetate extraction part; the ethyl acetate extraction part is subjected to silica gel column chromatography, acetone-petroleum ether is calculated by v / v, 0:100, 1:200, 1:100, 2:98, 5:95, 10:90, 20:80, 40:60, 50:50 as eluent, and gradient elution is performed to obtain 6 fractions Fr.1-Fr.6; the fraction Fr.4 is subjected to silica gel column chromatography, acetone-petroleum ether is calculated by v / v, 5:95, 10:90, 20:80, 40:60, 50:50 as eluent, and gradient elution is performed to obtain 6 fractions Fr.4.1-Fr.4.6, wherein the fraction Fr.4.4 is subjected to repeated MCI gel chromatography Compounds 1, 2, 3, 4, 5, 6, 7, and 8 are obtained by CHP P20 column chromatography, silica gel column chromatography, gel column chromatography, reverse phase column chromatography, and semi-preparative HPLC; then one of compounds 1-8 or any combination thereof is taken and a pharmaceutically acceptable carrier is added.

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