Terpene compounds, preparation methods and applications thereof

Abstract

The present invention belongs to the field of medical technology, and specifically, relates to a terpenoid compound, a preparation method and an application thereof. The terpenoid compound of the present invention includes Sinunarongter A and Sinunanolobatone A, the structural formula of Sinunarongter A is shown in Formula I, and the structural formula of Sinunanolobatone A is shown in Formula II: The terpenoid compound of the present invention can not only be used as a lead compound or a leading compound for the development of new anti-inflammatory drugs, but also can be used as a secondary metabolite to study the changes in the marine ecological environment.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular, relates to a terpene compound, a preparation method and application thereof. Background Art

[0002] my country has vast sea areas with many coral reefs and coral islands, which contain rich coral resources. It is one of the seas with concentrated distribution of soft corals in the world. In-depth and systematic research on the chemical composition and biological activity of such marine organisms is one of the important ways to discover new drug leads. Soft corals are the general name of the order of Anthozoa and the order of Soft Corals. They are marine coelenterates among invertebrates. They are mainly distributed in tropical and subtropical shallow seas, and a few are distributed in temperate, cold and deep sea areas. Among them, short-fingered soft corals are favored by researchers at home and abroad. Researchers have discovered a large number of new compounds in short-fingered soft corals, including terpenes, steroids, long-chain fatty acids, etc. Among them, there are novel structures and significant active ingredients, which show that soft corals have rich chemical diversity and biodiversity, and have potential application value in drug development.

[0003] Secondary metabolites are a class of small molecule organic compounds produced by secondary metabolism that are not essential for the normal operation of cellular life activities or plant growth and development. Biological secondary metabolites are an adaptation of organisms to the environment and are the result of the interaction with biological and non-biological factors in long-term evolution. Soft corals are rich in secondary metabolites, which are small molecule compounds that are not essential for cellular life activities and normal growth and development of organisms. Studying the secondary metabolites of soft corals can also provide ecologists with a way to study changes in the marine ecological environment.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] The primary purpose of the present invention is to provide two terpenoid compounds.

[0006] The second object of the present invention is to provide a method for preparing the terpenoid compound.

[0007] The third invention objective of the present invention is to propose the application of the terpene compound.

[0008] In order to achieve the purpose of the invention, the technical solution adopted is:

[0009] The present invention relates to a terpenoid compound, which is selected from the compounds represented by formula I or formula II:

[0010]

[0011] The present invention also relates to a method for preparing the terpenoid compound, which at least comprises the following steps:

[0012] S1, chopping short-fingered soft coral into pieces, and soaking and extracting with a first organic solvent to obtain an extract, wherein the first organic solvent is a ketone solvent;

[0013] S2, concentrating the extract to obtain a crude extract, distributing the crude extract between water and a second organic solvent in a volume ratio of 1:2-3, retaining the extract of the second organic solvent and concentrating it to obtain an organic phase extract, wherein the second organic solvent is selected from at least one of an ether solvent or an alcohol solvent;

[0014] S3, subjecting the organic phase extract to 200-300 mesh silica gel column chromatography, using petroleum ether-diethyl ether with a volume ratio of 100:0, 95:5, 8:2, 7:3, 6:4, 5:5, and 0:100 for gradient elution, with a flow rate of preferably 50 mL / min, collecting the elution fractions from 100:0 to 95:5 as component A, and collecting the elution fractions from 95:5 to 8:2 as component B;

[0015] S4, subjecting component A to Sephadex LH-20 gel column chromatography to obtain subcomponent A-1, and subjecting component B to Sephadex LH-20 gel column chromatography to obtain subcomponent B-1;

[0016] S5, subjecting the subcomponent A-1 to 300-400 silica gel column chromatography to obtain component A-2; subjecting the subcomponent B-1 to 300-400 silica gel column chromatography to obtain component B-2;

[0017] S6. The component A-2 is separated and purified by reversed-phase high performance liquid chromatography to obtain a compound of formula II; the component B-2 is separated and purified by reversed-phase high performance liquid chromatography to obtain a compound of formula I.

[0018] The invention also relates to the application of the terpenoid compound in the preparation of anti-inflammatory drugs.

[0019] The invention also relates to the application of the terpenoid compound as a secondary metabolite in the research of marine ecological environment.

[0020] The technical solution of the present invention has at least the following technical effects:

[0021] The terpenoid compound of the present invention is a new compound extracted from short-fingered soft coral (Sinularia nanolobata), or is used to provide a lead compound or a leading compound for the development of new anti-inflammatory drugs. In addition, it is used as a secondary metabolite of soft coral to study the changes in the marine ecological environment. DETAILED DESCRIPTION

[0022] In order to enable persons with ordinary knowledge in the art to understand the characteristics and effects of the present invention, the following is a general description and definition of the terms and expressions mentioned in the specification and the scope of the patent application. Unless otherwise specified, all technical and scientific words used in the text have the common meanings understood by those skilled in the art for the present invention. In case of conflict, the definitions in this specification shall prevail.

[0023] In the present invention, if the quantity or other numerical value or parameter is expressed as a range, a preferred range or a series of upper and lower limits, it should be understood that all ranges consisting of any upper limit or preferred value of the range and the lower limit or preferred value of the range have been specifically disclosed herein, regardless of whether these ranges are disclosed separately. In addition, if a range of numerical values ​​is mentioned herein, unless otherwise specified, the range should include its endpoints and all integers and fractions within the range. In the present invention, under the premise that the purpose of the invention can be achieved, the numerical value should be understood to have the accuracy of the effective digits of the numerical value. For example, the number 40.0 should be understood to cover the range from 39.50 to 40.49.

[0024] The following detailed description is merely illustrative in nature and is not intended to limit the present invention and its use. In addition, this document is not limited by any theory described in the above prior art or invention summary or the following detailed description or examples.

[0025] In a first aspect of an embodiment of the present invention, a terpenoid compound is provided, selected from Sinunarongter A and Sinunanolobatone A, wherein the structural formula of Sinunarongter A is shown in Formula I, and the structural formula of Sinunanolobatone A is shown in Formula II:

[0026]

[0027] The terpene compounds of the embodiments of the present invention are extracted from short-fingered soft coral (Sinularia nanolobata), or used to provide lead compounds or lead compounds for the development of new anti-inflammatory drugs. As secondary metabolites of soft corals, they can provide ecologists with a way to study changes in the marine ecological environment.

[0028] The second aspect of the embodiment of the present invention provides a method for preparing the terpene compound, which comprises at least the following steps:

[0029] S1, chopping short-finger soft coral (Sinularia nanolobata) into pieces, and soaking and extracting with a first organic solvent to obtain an extract;

[0030] S2, concentrating the extract to obtain a crude extract, distributing the crude extract between water and a second organic solvent, retaining the extract of the second organic solvent and concentrating it to obtain an organic phase extract,

[0031] S3. The organic phase extract is subjected to 200-300 mesh silica gel column chromatography, and gradient elution is performed with an eluent having a petroleum ether-to-ethyl ether volume ratio of (100:0, 95:5, 8:2, 7:3, 6:4, 5:5, 0:100), and the elution fractions from 100:0 to 95:5 are collected as component A, and the elution fractions from 95:5 to 8:2 are collected as component B; the flow rate is preferably 50 mL / min.

[0032] S4, subjecting component A to Sephadex LH-20 gel column chromatography to obtain subcomponent A-1, and subjecting component B to Sephadex LH-20 gel column chromatography to obtain subcomponent B-1;

[0033] S5, subjecting subcomponent A-1 to 300-400 silica gel column chromatography to obtain component A-2; subjecting subcomponent B-1 to 300-400 silica gel column chromatography to obtain component B-2;

[0034] S6. Component A-2 is separated and purified by reversed-phase high-performance liquid chromatography to obtain the terpene compound Sinunanolobatone A; component B-2 is separated and purified by reversed-phase high-performance liquid chromatography to obtain the new terpene compound Sinunarongter A.

[0035] As an implementation of the present invention, in S1, the first organic solvent is a ketone solvent, and preferably acetone. The first organic solvent is preferably used for immersion extraction 2 to 5 times to fully dissolve the organic components in the short-fingered soft coral.

[0036] As an implementation of the embodiment of the present invention, in S1, the immersion extraction is carried out under ultrasonic conditions; specifically, the ultrasonic conditions are a frequency of 53 kHz, a time of 15 minutes, and a water temperature of 25 to 30°C.

[0037] As an implementation of an embodiment of the present invention, in S2, the crude extract is distributed between water and a second organic solvent, i.e., the crude extract is extracted using the second organic solvent and water, wherein the volume ratio of water to the second organic solvent can be 1:2 to 3. The second organic solvent can be selected from at least one of an ether solvent or an alcohol solvent, specifically, the ether solvent can be selected from diethyl ether, and the alcohol solvent can be selected from n-butanol.

[0038] As an implementation of an embodiment of the present invention, in S2, the concentration may be performed by a commonly used concentration method in the art, and specifically may be performed under reduced pressure conditions.

[0039] As an implementation of an embodiment of the present invention, step S3 is column chromatography, the purpose of which is gradient elution to obtain an eluent component containing the target component for further purification. During the research process, the components of each gradient-changing eluent were analyzed, and it was found that the elution fractions from 100:0 to 95:5 and the elution fractions from 95:5 to 8:2 contained new compounds, and the content was relatively high, which can be used as component A and component B for further purification, respectively. Specifically, the silica gel column I is 200-300 mesh, which can achieve a good crude separation effect.

[0040] As an implementation of an embodiment of the present invention, component A and component B are purified by steps S4, S5, and S6, respectively, so as to obtain target compounds Sinunanolobatone A and Sinunarongter A, respectively.

[0041] In S4, the gel column chromatography is selected from Sephadex LH-20 gel column, the eluent of the gel column chromatography is selected from the eluent with a volume ratio of petroleum ether: dichloromethane: methanol of 2:1:1, and the flow rate is preferably 3 mL / min.

[0042] In S5, the silica gel column is 300-400 mesh, which can achieve a better separation effect of each component. The gradient eluent of subcomponent A-1 is selected from petroleum ether-diethyl ether with a volume ratio of 100:0 to 100:1; the flow rate is preferably 20 mL / min. The gradient eluent of subcomponent B-1 is selected from petroleum ether-diethyl ether with a volume ratio of 90:5 to 90:9, and the flow rate is preferably 20 mL / min.

[0043] In S6, a Dikma Diamonsil Plus HPLC column is used in the reverse phase high performance liquid chromatography; wherein, component A-2 is isocratically eluted using an eluent having a volume ratio of acetonitrile:water of 8:2; and the flow rate is preferably 1.0 mL / min.

[0044] More preferably, component B-2 is isocratically eluted using an eluent having a volume ratio of acetonitrile to water of 7:3, and the flow rate is preferably 1.0 mL / min.

[0045] As an implementation method of an embodiment of the present invention, the method for separating terpenoid compounds comprises the following steps:

[0046] (1) Extraction: mince the short-fingered soft coral Sinularia nanolobata, soak it in acetone and perform ultrasonic extraction for 2-5 times to obtain an extract;

[0047] (2) Extraction: The extract in step (1) is concentrated to obtain a crude extract, the crude extract is extracted with ether or n-butanol, the organic extract is retained and concentrated under reduced pressure to obtain an organic phase extract;

[0048] (3) Column chromatography: The organic phase extract was subjected to silica gel column chromatography, and gradient elution was performed with petroleum ether-diethyl ether (100:0, 95:5, 8:2, 7:3, 6:4, 5:5, and 0:100, respectively). The elution fractions from 100:0 to 95:5 were collected as component A, and the elution fractions from 95:5 to 8:2 were collected as component B;

[0049] (4) Component A was subjected to gel column chromatography (Sephadex LH-20 gel column) to obtain subcomponent A-1, and component B was subjected to gel column chromatography (Sephadex LH-20 gel column) to obtain subcomponent B-1; the flow rate was 3 mL / min;

[0050] (5) Subcomponent A-1 was subjected to silica gel column chromatography, and the fraction was eluted with a gradient of petroleum ether to diethyl ether in a volume ratio of 100:0 to 100:1 as component A-2; subcomponent B-1 was subjected to silica gel column chromatography, and the fraction was eluted with a gradient of petroleum ether to diethyl ether in a volume ratio of 90:5 to 90:9 as component B-2; the flow rate was 20 mL / min;

[0051] (6) Component A-2 was separated and purified by reverse phase high performance liquid chromatography (Dikma Diamonsil Plus HPLC column, 5 μm C18-A, 250×4.6 mm column), eluted with an acetonitrile: water volume ratio of 8:2 at a flow rate of 1.0 mL / min, to obtain a new terpene skeleton compound Sinunanolobatone A; component B-2 was separated and purified by reverse phase high performance liquid chromatography (Dikma Diamonsil Plus HPLC column, 5 μm C18-A, 250×4.6 mm column), eluted with an acetonitrile: water volume ratio of 7:3 at a flow rate of 1.0 mL / min, to obtain a new terpene compound Sinunarongter A.

[0052] The third aspect of the embodiments of the present invention relates to the use of the terpene compound in the preparation of anti-inflammatory drugs. Experiments have found that the terpene compound has a certain anti-inflammatory effect and can be used to provide a lead compound or a lead compound for the development of new anti-inflammatory drugs.

[0053] The fourth aspect of the embodiments of the present invention relates to the application of the terpenoid compound as a secondary metabolite in the study of marine ecological environment. Secondary metabolites are small molecule compounds that are not essential for cell life activities and normal growth and development of organisms. The terpenoid compound of the present invention can provide a way to study changes in the marine ecological environment.

[0054] The present invention will be further described below in conjunction with embodiments, but the present invention is not limited thereto.

[0055] Example

[0056] The reagents and equipment used include:

[0057] Acetone, anhydrous ethyl ether, petroleum ether: Sinopharm Chemical Reagent Co., Ltd., China;

[0058] 200-300 mesh silica gel: Santai Technology Co., Ltd., China;

[0059] 300-400 mesh silica gel: Sinopharm Chemical Reagent Co., Ltd., China;

[0060] Dikma Diamonsil Plus HPLC column (5 μm C18-A, 250 × 4.6 mm): Dikma Technology;

[0061] Agilent 1260 detector: Agilent Technologies; Bruker DRX-500 spectrometer: Bruker Biospin AG, Germany.

[0062] Example 1 Preparation method of terpenoid compound Sinunanolobatone A

[0063] (1) The short-finger soft coral (Sinularia nanolobata) sample was collected from the waters of Xidao Island, Hainan, China in 2017. After collection, it was frozen and extracted with acetone five times;

[0064] (2) combining the extracts, concentrating under reduced pressure to obtain a crude extract, distributing the extract between water and a second organic solvent, anhydrous ether, in a volume ratio of 1:2 to 3, extracting the crude extract until the supernatant is colorless, and concentrating under reduced pressure to obtain an anhydrous ether extract;

[0065] (3) The anhydrous ether extract was subjected to silica gel (200-300 mesh) column chromatography, and gradient elution was performed with an organic solvent of petroleum ether-ether (100:0, 95:5, 8:2, 7:3, 6:4, 5:5, 0:100, in sequence), and the elution fractions from 100:0 to 95:5 (component A) were collected; the flow rate was 50 mL / min;

[0066] (4) The collected elution fractions were subjected to gel column chromatography (Sephadex LH-20) using an eluent having a volume ratio of petroleum ether: dichloromethane: methanol of about 2:1:1 to obtain subfraction A-1; the flow rate was 3 mL / min;

[0067] (5) subjecting subcomponent A-1 to silica gel (300-400 mesh) column chromatography, using an organic solvent of petroleum ether-diethyl ether (volume ratio 100:0 to 100:1) for gradient elution, and collecting the eluted fractions (component A-2); the flow rate is 20 mL / min;

[0068] (6) A-2 was separated and purified by reverse phase high performance liquid chromatography (Dikma Diamonsil Plus HPLC column, 5 μm C18-A, 250×4.6 mm) (acetonitrile: water = 8:2, 1.0 mL / min) to obtain 1.0 mg of the terpene compound Sinunanolobatone A.

[0069] The above terpenoid compounds have the following structure after spectral analysis:

[0070]

[0071] Sinunanolobatone A structure and physicochemical properties of the compound:

[0072] 1 H NMR data: d H 0.50(1H,dd,J=3.7,6.5Hz),1.39(3H,s),1.54(3H,s),1.59(3H,s),1.63(3H,s),1.75(1H,m),1 .83(1H,dd,J=3.7,9.2Hz),1.93(1H,m),2.01(1H,m),2.06(1H,m),2.11(1H,m),2.16(1H,m),2. 16(1H,m),2.17(1H,m),2.24(1H,m),2.34(1H,d,J=4.6Hz),2.36(1H,m),2.45(1H,m),2.85(1H, ddd), 4.90 (1H, d, J = 8.2Hz), 4.97 (1H, dd, J = 3.6, 9.5Hz), 5.14 (1H, ddt, J = 1.3, 2.7, 5.3Hz) ppm; 13 C NMR data: d C 15.5 (C-18), 15.6 (C-19), 15.8 (C-17), 16.7 (C-20), 22.9 (C-16), 23.9 (C-10), 25.1 (C-6), 30.8 (C-1), 31.3 (C-2), 36.5 (C-13), 38.0 (C-14), 39.0 (C-5), 39.0 (C-9), 123.1 (C-13), 125.3 (C-11), 125.4 (C-7), 133.7 (C-12), 134.4 (C-8), 138.8 (C-4), 213.6 (C-15) ppm. Mass spectral data: HR-EIMS: m / z 286.2282 [M] + (C 20 H 30O, calculated value 286.2291).

[0073] NMR test with CDCl 3 (δ H 7.26ppm,δ C 77.2ppm) as internal standard.

[0074] Example 2 Preparation method of terpenoid compound Sinunarongter A

[0075] (1) The short-finger soft coral (Sinularia nanolobata) sample was collected from the waters of Xidao Island, Hainan, China in 2017. After collection, it was frozen and extracted with acetone five times;

[0076] (2) combining the extracts, concentrating under reduced pressure to obtain a crude extract, distributing the extract between water and a second organic solvent, anhydrous ether, in a volume ratio of 1:2 to 3, extracting the crude extract until the supernatant is colorless, and concentrating under reduced pressure to obtain an anhydrous ether extract;

[0077] (3) The anhydrous ether extract was subjected to silica gel (200-300 mesh) column chromatography, and gradient elution was performed with an organic solvent of petroleum ether-ether (100:0, 95:5, 8:2, 7:3, 6:4, 5:5, 0:100, in sequence), and the 95:5 to 8:2 elution fractions (component B) were collected; the flow rate was 50 mL / min;

[0078] (4) The collected elution fractions were subjected to gel column chromatography (Sephadex LH-20) using an eluent having a volume ratio of petroleum ether: dichloromethane: methanol of 2:1:1 to obtain subfraction B-1; the flow rate was 3 mL / min;

[0079] (5) subjecting subcomponent B-1 to silica gel (300-400 mesh) column chromatography, using an organic solvent of petroleum ether-diethyl ether (volume ratio 90:5 to 90:9) for gradient elution, and collecting the eluted fraction (component B-2); the flow rate was 20 mL / min;

[0080] (6) B-2 was separated and purified by reverse phase high performance liquid chromatography (Agilent 1260 detector, Dikma Diamonsil Plus HPLC column, 5 μm C18-A, 250×4.6 mm) (acetonitrile: water = 7:3, 1.0 mL / min) to obtain 11.3 mg of the terpene compound Sinunarongter A.

[0081] The above terpenoid compounds have the following structure after spectral analysis:

[0082]

[0083] The physical and chemical properties of the compound Sinunarongter A:

[0084] 1 H NMR data: d H 0.81(1H,m),0.93(3H,d,J=6.8Hz),1.15(1H,m),1.19(3H,s),1.31(1H,m),1.50(1H, m),1.56(1H,m),1.67(3H,s),1.70(3H,s),1.99(1H,m),2.16(1H,m),2.17(1H,m),2. 19(1H,m),2.21(1H,m),2.22(1H,m),2.27(1H,m),2.43(1H,dd,J=7.7,15.4Hz),2.89 (1H,d,J=13.4Hz),3.05(1H,d,J=13.4Hz),3.64(3H,s),4.77(1H,m),5.11(1H,m)ppm; 13 C NMR data: d C 9.6 (C-16), 16.7 (C-18), 17.4 (C-19), 21.3 (C-20), 23.0 (C-14), 25.1 (C-6), 27.4 (C-15), 28.3 (C-2), 30.0 (C-12), 31.3 (C-1), 37.4 (C-13), 39.0 (C-5), 50.9 (C-11), 52.1 (C-21), 52.8 (C-9), 117.7 (C-3), 129.3 (C-7), 129.6 (C-8), 138.6 (C-4), 176.8 (C-17), 209.8 (C-10) ppm. Mass spectral data: HR-ESIMS: m / z 333.2428 [M+H] + (C 21 H 32 O 3 , calculated value 333.2424).

[0085] NMR test with CDCl 3 (δ H 7.26ppm,δ C 77.2ppm) as internal standard.

[0086] Example 3 Experiment on the improvement of lipopolysaccharide-induced BV2 cell inflammation

[0087] 1. Experimental Principle

[0088] Lipopolysaccharide (LPS)-induced BV2 cell inflammation model

[0089] Microglia are widely distributed in the central nervous system. They are the main cells that play an immune function in the nervous system. Under physiological conditions, they play a monitoring role and stabilize the internal environment. Microglia are very sensitive to external stimuli. Once a slight pathological change occurs in the central nervous system, microglia can be activated. On the one hand, the activated microglia play a phagocytic role, engulf cell fragments, and secrete growth factors to promote nerve repair. On the other hand, they can also secrete some inflammatory factors, glutamate, nitric oxide, etc., which aggravate the inflammatory response and cause secondary damage. The experiment used 100ng / mL LPS to stimulate microglia and release a large number of inflammatory factors, including NO. The Griess reagent method was used to detect the content of NO in the supernatant to preliminarily judge the level of microglial inflammatory response.

[0090] 2. Experimental Methods

[0091] 1. Lipopolysaccharide (LPS)-induced BV2 cell inflammation model

[0092] (1) BV2 cells (purchased from ATCC) were digested with 0.25% trypsin and suspended in DMEM high-glucose culture medium containing 10% fetal bovine serum.

[0093] (2) 2×10 5 BV2 cells were inoculated in a 96-well culture plate at a density of 100 μL / well and placed in a 5% CO 2 Cultured in a 37°C constant temperature incubator.

[0094] (3) After culturing BV2 cells for 24 hours, the culture medium of each group was replaced with fresh DMEM high-glucose culture medium containing 10% fetal bovine serum.

[0095] (4) The test compound of corresponding concentration was added to the drug-treated group (10 μL / well), and DMEM high-glucose culture medium containing 10% fetal bovine serum (10 μL / well) was added to the normal control group and LPS model group.

[0096] (5) After 2 hours of incubation, 0.001 mg / mL LPS (10 μL / well) was added to the drug administration group and LPS model group, respectively, with a final concentration of 100 ng / mL. DMEM high-glucose culture medium containing 10% fetal bovine serum (10 μL / well) was added to the normal control group.

[0097] (6) After culturing for 24 hours, 50 μL of supernatant and 50 μL of Greiss buffer were taken from each well and reacted at room temperature for 15 min. The OD value of each group was measured at a wavelength of 540 nm.

[0098] 2. Effects of compounds on cell viability

[0099] (1) BV2 cells were digested with 0.25% trypsin and suspended in DMEM high-glucose culture medium containing 10% fetal bovine serum.

[0100] (2) 1.25×10 4 BV2 cells were inoculated in a 96-well culture plate at a density of 100 μL / well and placed in a 5% CO 2 Cultured in a 37°C constant temperature incubator.

[0101] (3) After culturing BV2 cells for 24 hours, the culture medium of each group was replaced with fresh DMEM high-glucose culture medium containing 10% fetal bovine serum.

[0102] (4) 20 μM of the test compound (10 μL / well) was added to the drug-treated group, 20 μM resveratrol (10 μL / well) was added to the positive control group, and DMEM high-glucose culture medium containing 10% fetal bovine serum (10 μL / well) was added to the normal control group.

[0103] (5) After culturing for 24 hours, 5 mg / ml MTT (10 μL / well) was added for live cell staining.

[0104] (6) After incubation for 3 hours, the culture medium was discarded, 100% DMSO (100 μL / well) was added, and the cells were shaken on a plate shaker to fully dissolve.

[0105] (7) The OD value of each group was measured at a wavelength of 490 nm.

[0106] 3. Experimental Conclusion

[0107] The NO inhibition rate of Sinunanolobatone A at 20 μM was 70.54±1.44%, and the NO inhibition rate of Sinunarongter A at 20 μM was 85.93±4.35%, both of which were higher than the inhibition rate of the positive control resveratrol at 20 μM (69.38±7.76%) on LPS-induced BV2 cell inflammation. And at 20 μM, compared with the positive control resveratrol (cell survival rate 54.58±6.30%), Sinunanolobatone A (cell survival rate 98.05±0.88%) and Sinunarongter A (cell survival rate 96.14±2.29%) had no obvious cytotoxicity.

[0108] The above embodiments are essentially only for auxiliary explanation and are not intended to limit the embodiments of the application target or the application or use of these embodiments. In this article, the term "exemplary" means "as an example, example or illustration". Any exemplary embodiment in this article is not necessarily interpreted as being preferred or more advantageous than other embodiments.

[0109] In addition, although at least one exemplary embodiment or comparative example has been proposed in the aforementioned embodiments, it should be understood that the present invention can still have a large number of changes. It should also be understood that the embodiments described herein are not intended to limit the scope, use or configuration of the requested application target by any means. On the contrary, the aforementioned embodiments will provide a simple guide for those with ordinary knowledge in the art to implement one or more of the embodiments described. Furthermore, various changes can be made to the functions and arrangements of the elements without departing from the scope defined by the scope of the application, and the scope of the application includes known equivalents and all foreseeable equivalents when the present patent application is filed.

Claims

1. A terpenoid compound, which is selected from the compounds represented by formula I or formula II:

2. The method for preparing terpenoid compounds according to claim 1, characterized in that: At least the following steps are included: S1, chopping short-fingered soft coral into pieces, and soaking and extracting with a first organic solvent to obtain an extract, wherein the first organic solvent is a ketone solvent; S2, concentrating the extract to obtain a crude extract, distributing the crude extract between water and a second organic solvent in a volume ratio of 1:2-3, retaining the extract of the second organic solvent and concentrating it to obtain an organic phase extract, wherein the second organic solvent is selected from at least one of an ether solvent or an alcohol solvent; S3, subjecting the organic phase extract to 200-300 mesh silica gel column chromatography, gradient elution with petroleum ether-diethyl ether in volume ratios of 100:0, 95:5, 8:2, 7:3, 6:4, 5:5, and 0:100, collecting the elution fractions from 100:0 to 95:5 as component A, and collecting the elution fractions from 95:5 to 8:2 as component B; S4, subjecting component A to Sephadex LH-20 gel column chromatography to obtain subcomponent A-1, and subjecting component B to Sephadex LH-20 gel column chromatography to obtain subcomponent B-1; S5, subjecting the subcomponent A-1 to 300-400 silica gel column chromatography to obtain component A-2; subjecting the subcomponent B-1 to 300-400 silica gel column chromatography to obtain component B-2; S6. The component A-2 is separated and purified by reverse phase high performance liquid chromatography to obtain a compound represented by formula II; The component B-2 is separated and purified by reverse phase high performance liquid chromatography to obtain a compound shown in formula I.

3. The preparation method according to claim 2, characterized in that: In S1, the first organic solvent is acetone.

4. The preparation method according to claim 2, characterized in that: In S1, the first organic solvent is used for immersion extraction 2 to 5 times.

5. The preparation method according to claim 2, characterized in that: In S1, the soaking extraction is performed under ultrasonic conditions.

6. The preparation method according to claim 2, characterized in that: In S2, the ether solvent is diethyl ether, and the alcohol solvent is n-butanol.

7. The preparation method according to claim 2, characterized in that: In S2, the concentration is performed under reduced pressure.

8. The preparation method according to claim 2, characterized in that: In S3, the flow rate of the gradient elution is 50 mL / min.

9. The preparation method according to claim 2, characterized in that: In S4, the eluent of the gel column chromatography is selected from an eluent having a volume ratio of petroleum ether: dichloromethane: methanol of 2:1:

1.

10. The preparation method according to claim 9, characterized in that: The flow rate was 3 mL / min.

11. The preparation method according to claim 2, characterized in that: In S5, the gradient eluent of the subcomponent A-1 is selected from petroleum ether-ethyl ether in a volume ratio of 100:0 to 100:

1.

12. The preparation method according to claim 11, characterized in that: The flow rate is 20 mL / min.

13. The preparation method according to claim 2, characterized in that: In S5, the gradient eluent of the subcomponent B-1 is selected from petroleum ether-ethyl ether in a volume ratio of 90:5 to 90:

9.

14. The preparation method according to claim 13, characterized in that: The flow rate is 20 mL / min.

15. The preparation method according to claim 2, characterized in that: In S6, the reverse phase high performance liquid chromatography uses a Dikma Diamonsil Plus HPLC chromatography column.

16. The preparation method according to claim 15, characterized in that: The component A-2 is isocratically eluted using an eluent having a volume ratio of acetonitrile to water of 8:2; The component B-2 was isocratically eluted using an eluent having a volume ratio of acetonitrile to water of 7:

3.

17. Use of the terpenoid compound according to claim 1 in the preparation of anti-inflammatory drugs.

Images