Novel diterpenoid glycoside compound, preparation method and application

Abstract

The invention provides a novel diterpene glycoside compound as well as a preparation method and application thereof, and belongs to the technical field of medical biology. The novel diterpenoid glycoside compound is extracted and separated from Lemnalia sarcophyllum for the first time, and the molecular structure, the separation method and the pharmacological activity research of the novel diterpenoid glycoside compound are not reported by the existing journal paper. The diterpene glycoside novel compound obtained through separation and purification has an anti-tumor effect, has inhibitory activity on a drug-resistant human small cell lung cancer cell strain NCI-H446 / EP, can be used for developing candidate molecules for treating the disease, and can also be used as a lead compound for chemical synthesis and structural modification of other drugs. The preparation method provided by the invention is simple and rapid, and the extracted new diterpene glycoside compound is high in purity.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and relates to a novel diterpene glycoside compound, a preparation method and an application thereof. Background Art

[0002] Compared with terrestrial organisms, the living environment of marine organisms has characteristics such as high salinity, high pressure, low oxygen, and low light, covering various ecological environments such as the intertidal zone, polar regions, deep-sea abysses, hydrothermal vents, and cold seeps. This has shaped the particularity of marine organisms in terms of metabolism, survival methods, information transmission, and adaptation mechanisms.

[0003] Soft corals are marine invertebrates of lower order. They are numerous in quantity and variety, and the secondary metabolites produced thereby are also diverse, having great potential for marine medicinal development. Lemnalia Genus Sinularia is widely distributed in the marine ecosystem. Its main secondary metabolites are terpene compounds, including sesquiterpenes, diterpenes, and diterpene glycoside compounds. These secondary metabolites have good and diverse biological activities, involving antibacterial, antiviral, neuroprotective, and cytotoxic activities. Therefore, the research on marine organisms including soft corals can obtain natural products with diverse structures, discover monomeric compounds with good biological activities, provide valuable candidate molecules for the development and synthesis of drugs, and also provide a scientific basis for the rational development and utilization of marine biological resources.

[0004] Lung cancer is a common malignant tumor that seriously threatens human health. According to histological characteristics, it can be divided into non-small cell lung cancer and small cell lung cancer. Small cell lung cancer is a pulmonary neuroendocrine tumor with rapid growth and easy metastasis, and has an extremely high mortality rate. At present, the clinical treatment of small cell lung cancer generally adopts a method combining radiotherapy and chemotherapy. This method usually responds well in the initial stage of treatment, but patients quickly develop drug resistance and relapse. The clinical tests of early developed targeted drugs are not very satisfactory, and there is an urgent need for new targeted drugs and effective treatment means for small cell lung cancer. Summary of the Invention

[0005] The purpose of the present invention is to provide a novel diterpene glycoside compound, a preparation method and an application thereof, so as to solve the problems of easy drug resistance and easy recurrence of the existing treatment methods for small cell lung cancer.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions: In the first aspect, the present application provides a novel diterpene glycoside compound, which is extracted from soft corals, especially Lemnalia Sinularia. The novel diterpene glycoside compound in the present application is named lemnaliside A , which is a colorless transparent oily substance, with a molecular formula of C 26 H 36 O6, and the molecular structural formula is: 。

[0007] In a second aspect, the present application provides a method for preparing a new diterpene glycoside compound lemnaliside A , and the method includes: S01: The soft coral is shredded and soaked in methanol at room temperature to obtain an extract. After the extract is concentrated under reduced pressure, desalted with anhydrous methanol, and concentrated under reduced pressure again, a crude extract is obtained.

[0008] Thawed the cryopreserved Lemnalia genus of soft coral at room temperature and shredded it into pieces the size of a fingernail. The shredded soft coral was soaked in methanol at room temperature 6 times, each time for 3 days. After the soaking, the filtrate was filtered and all the soaking solutions were combined to obtain an extract. The extract was concentrated under reduced pressure to a paste at a temperature of 37°C and a pressure of 0.1 MPa. Anhydrous methanol was added to the concentrated extract to redissolve it, and after desalting by filtration, it was concentrated under reduced pressure at a temperature of 37°C and a pressure of 0.1 MPa to obtain a crude extract.

[0009] S02: After the crude extract is separated by silica gel vacuum column chromatography, gradient elution, and thin-layer chromatography color development detection, the same fractions are combined and concentrated to obtain 16 primary components F1 - F16.

[0010] The crude extract was chromatographically separated using a silica gel vacuum column with a column specification of 160 mm * 500 mm and a silica gel particle size of 200 - 300 mesh. The separated products were successively gradient eluted with petroleum ether, petroleum ether / acetone with volume ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, dichloromethane, dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1. The eluted fractions were subjected to thin-layer chromatography color development detection. The fractions with the same fractions were combined and concentrated to obtain 16 primary components F1 - F16. Among them, the thin-layer chromatography color development detection conditions were: the developing agent was petroleum ether and propanol with a volume ratio of 3:1, and the coloring agent was a sulfuric acid ethanol solution with a mass concentration of 10%.

[0011] S03: After the F16 among the primary components F1 - F16 is separated by silica gel column chromatography, gradient elution, and thin-layer chromatography color development detection, the same fractions are combined and concentrated to obtain 6 secondary components F161 - F166.

[0012] The component F16 in the primary components F1 - F16 was chromatographically separated using a silica gel column with a column specification of 60 mm * 500 mm and a silica gel particle size of 300 - 400 mesh. The separation products were successively gradient eluted with dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, and 1 / 1. The eluted components were detected by thin-layer chromatography color development, and the components with the same fractions were combined and concentrated to obtain 6 secondary components F161 - F166. Among them, the thin-layer chromatography color development detection conditions were: the developing agent was petroleum ether and propanol with a volume ratio of 3:1, and the color developing agent was a sulfuric acid ethanol solution with a mass concentration of 10%.

[0013] S04: After high-performance liquid chromatography analysis, medium-pressure preparative liquid chromatography separation, and isocratic elution of F165 among the secondary components F161 - F166, 4 tertiary components F1651 - F1654 were obtained.

[0014] Perform high-performance liquid chromatography analysis on the component F165 in the secondary components F161 - F166 to determine that the separation and preparation chromatographic conditions are methanol / water with a volume ratio of 80 / 20. Among them, the high-performance liquid chromatography analysis conditions are: the analytical chromatographic column is a Greenherb ODS C column with a size of 4.6 mm * 250 mm and a particle size of 5 μm 18 column, and the flow rate is 1 ml / min. Separate the component F165 using a medium-pressure preparative liquid chromatograph, and the separation products are isocratically eluted with methanol / water with a volume ratio of 80 / 20 to obtain 4 tertiary components F1651 - F1654. According to the retention time in the high-performance liquid chromatography chart of the component F165, it is determined that: 0.00 - 7.60 min is the component F1651; 7.60 - 12.50 min is the component F1652; 12.50 - 17.50 min is the component F1653; 17.50 - 25.00 min is the component F1654. Among them, the medium-pressure preparative liquid chromatography separation conditions are: the preparative chromatographic column is a Greenherb ODS C column with a size of 20 mm * 250 mm and a particle size of 5 μm 18 column, and the flow rate is 10 ml / min.

[0015] S05: After high-performance liquid chromatography analysis, high-performance preparative liquid chromatography separation, and isocratic elution of F1654 among the tertiary components F1651 - F1654, a new diterpene glycoside compound was obtained.

[0016] Perform high-performance liquid chromatography purification analysis on the component F1654 in the tertiary components F1651 - F1654 to determine that the separation and preparation chromatographic conditions are acetonitrile / water with a volume ratio of 35 / 65. Among them, the high-performance liquid chromatography analysis conditions are: the analytical chromatographic column is a Greenherb ODS C column with a size of 4.6 mm * 250 mm and a particle size of 5 μm 18Column, flow rate of 1 ml / min. The component F1654 was separated and purified by high-performance preparative liquid chromatography, and the separated product was isocratically eluted with acetonitrile / water with a volume ratio of 35 / 65 at a retention time of 127 min to obtain a new diterpene glycoside compound lemnaliside A . Among them, the separation conditions of high-performance preparative liquid chromatography are as follows: the chromatographic column is a Greenherbs ODS C column with a size of 10 mm * 250 mm and a particle size of 5 μm 18 , and the flow rate is 2 ml / min.

[0017] Thirdly, the new diterpene glycoside compound prepared in this application lemnaliside A is used to prepare a drug for treating drug-resistant human small cell lung cancer.

[0018] The present invention has the following beneficial effects: (1) For the first time in this application, a new diterpene glycoside compound was extracted and separated from soft corals of the genus Lemnalia , and the molecular structure, separation method, and pharmacological activity of this new diterpene glycoside compound lemnaliside A have not been reported in existing journal papers. lemnaliside A (2) This new diterpene glycoside compound

[0019] has an anti-tumor effect, has inhibitory activity against the drug-resistant human small cell lung cancer cell line NCI-H446 / EP, can be used to develop candidate molecules for treating this disease, and can also be used as a lead compound for other pharmaceutical chemical synthesis and structural modification. lemnaliside A (3) The preparation method of this new diterpene glycoside compound

[0020] is simple and fast, and the purity of the extracted new diterpene glycoside compound is high. lemnaliside A (4) The UV (Ultraviolet Visible Absorption Spectroscopy) diagram of the new diterpene glycoside compound prepared in Example 1 of this application Description of the drawings

[0021] Figure 1 is the high-performance liquid analysis chromatogram of the F165 component during the separation process of the new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A ; Figure 2 is the UV (Ultraviolet Visible Absorption Spectroscopy) diagram of the new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A ; Figure 3 is the IR (Infrared Spectroscopy) diagram of the new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A ; Figure 4 is the NMR (Nuclear Magnetic Resonance) spectrum diagram of the new diterpene glycoside compound prepared in Example 1 of this application lemnaliside AHRESIMS (high resolution electrospray ionization mass spectroscopy) spectrum; Figure 5 is a new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A of 1 1H-NMR (Nuclear Magnetic Resonance Spectroscopy of Hydrogen) spectrum; Figure 6 is a new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A of 13 13C-NMR (Carbon-13 nuclear magnetic resonance) spectrum; Figure 7 is a new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A HSQC (heteronuclear singular quantum correlation) spectrum; Figure 8 is a new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A HMBC (1H detected heteronuclear multiple bond correlation) spectrum; Figure 9 is a new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A 1H-1H COSY (Homonuclear Chemical Shift Correlation Spectroscopy) spectrum; Figure 10 is a new diterpene glycoside compound prepared in Example 1 of this application lemnaliside A Comparison chart of TDDFT-ECD calculation and experimental CD values; Detailed implementation method

[0022] The technical solutions of the present invention will be further explained and illustrated through specific examples below.

[0023] Example 1 The example of this application provides a new diterpene glycoside compound lemnaliside A which lemnaliside A is a colorless transparent oil, and its molecular formula is C26 H 36 O6, with the molecular structural formula as follows: .

[0024] Example 2 The embodiment of the present application provides a preparation method of a new diterpene glycoside compound lemnaliside A which includes: S201: Thaw the frozen Lemnalia soft coral at room temperature and cut it into pieces the size of a fingernail. Immerse the cut soft coral in normal-temperature methanol 6 times, with each immersion lasting for 3 days. After the immersion, filter and combine all the immersion liquids to obtain an extract. Concentrate the extract under reduced pressure at a temperature of 37°C and a pressure of 0.1 MPa until it becomes a paste. Add anhydrous methanol to the concentrated paste extract to redissolve it, filter to remove salts, and then concentrate it under reduced pressure at a temperature of 37°C and a pressure of 0.1 MPa to obtain a crude extract.

[0025] S202: Perform column chromatography separation on the crude extract using a silica gel decompression column with a column chromatography specification of 160 mm * 500 mm and a silica gel particle size of 200 - 300 mesh. The separated products are eluted successively with petroleum ether, petroleum ether / acetone with volume ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, dichloromethane, and dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1 for gradient elution. The obtained elution components are subjected to thin-layer chromatography color development detection, and the components with the same fractions are combined and concentrated to obtain 16 primary components F1 - F16. Among them, the thin-layer chromatography color development detection conditions are: the developing agent is petroleum ether and propanol with a volume ratio of 3:1, and the color-developing agent is a sulfuric acid ethanol solution with a mass concentration of 10%.

[0026] S203: Perform chromatographic separation on component F16 among the primary components F1 - F16 using a silica gel column with a column chromatography specification of 60 mm * 500 mm and a silica gel particle size of 300 - 400 mesh. The separated products are eluted successively with dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, 1 / 1 for gradient elution. The elution components are subjected to thin-layer chromatography color development detection, and the components with the same fractions are combined and concentrated to obtain 6 secondary components F161 - F166. Among them, the thin-layer chromatography color development detection conditions are: the developing agent is petroleum ether and propanol with a volume ratio of 3:1, and the color-developing agent is a sulfuric acid ethanol solution with a mass concentration of 10%.

[0027] S204: Perform high-performance liquid chromatography analysis on component F165 among the secondary components F161 - F166 to determine that the separation and preparation chromatographic conditions are methanol / water with a volume ratio of 80 / 20, and obtain Figure 1The high-performance liquid chromatography (HPLC) chromatogram shown, where the HPLC analysis conditions are as follows: the analytical chromatographic column is a Green Herb ODS C column with dimensions of 4.6 mm * 250 mm and a particle size of 5 μm. 18 The column flow rate is 1 ml / min. According to the attached Figure 1 Based on the HPLC chromatogram shown, a medium-pressure preparative liquid chromatograph was used to separate component F165. The separated product was isocratically eluted with methanol / water at a volume ratio of 80 / 20, yielding four tertiary components F1651 - F1654. Specifically, based on the retention time, the four tertiary components F1651 - F1654 were determined as follows: 0.00 - 7.60 min was component F1651; 7.60 - 12.50 min was component F1652; 12.50 - 17.50 min was component F1653; 17.50 - 25.00 min was component F1654. Among them, the medium-pressure preparative liquid chromatography separation conditions were: the preparative chromatographic column was a Green Herb ODS C column with dimensions of 20 mm * 250 mm and a particle size of 5 μm. 18 The column flow rate was 10 ml / min.

[0028] S205: Component F1654 in the tertiary components F1651 - F1654 was subjected to HPLC purification analysis to determine that the separation and preparation chromatographic conditions were acetonitrile / water at a volume ratio of 35 / 65. Among them, the HPLC analysis conditions were: the analytical chromatographic column was a Green Herb ODS C column with dimensions of 4.6 mm * 250 mm and a particle size of 5 μm. 18 The column flow rate was 1 ml / min. Component F1654 was separated and purified using a high-performance preparative liquid chromatograph. The separated product was isocratically eluted with acetonitrile / water at a volume ratio of 35 / 65 at a retention time of 127 min, yielding a new diterpene glycoside compound. lemnaliside A Among them, the high-performance preparative liquid chromatography separation conditions were: the chromatographic column was a Green Herb ODS C column with dimensions of 10 mm * 250 mm and a particle size of 5 μm. 18 The column flow rate was 2 ml / min.

[0029] Example 3 To determine the structure of the new diterpene glycoside compound obtained by the preparation method provided in Example 2, the new diterpene glycoside compound extracted in Example 2 in this application of the present application lemnaliside A was respectively subjected to ultraviolet spectroscopy, infrared spectroscopy, high-resolution mass spectrometry, lemnaliside A 1H-NMR, 1 13C-NMR, HSQC, HMBC, 13 1H- 1 1H COSY detections, and ECD calculations were performed to determine its absolute configuration, obtaining the attached 1 and the one-dimensional and two-dimensional nuclear magnetic resonance data shown in Table 1. Figures 2 - 10 ​

[0030] From the attached Figures 2 - 4 It can be seen that the new diterpene glycoside compounds lemnaliside A In α 25 D -30.1 ( c 1.0, MeOH); ECD ( c 0.5, MeOH) = Δ ε 195 -1.56, Δ ε 208 –0.81, Δ ε 224 +4.45; UV (MeOH) λ max (log ε ) = 231(1.78) nm; IR (KBr) ν max = 3421, 1631, 1592, 1369 cm -1 ; HRESIMS m / z 489.2499[M + CH2O2 - H] - , the calculated value is C 27 H 37 O8, 489.2494, which indicates that the molecular formula of this new compound is C 26 H 36 O6, and the degree of unsaturation is 6.

[0031] Table 1: One-dimensional and two-dimensional nuclear magnetic resonance data of the new diterpene glycoside compounds lemnaliside A (CDOD3, 500 MHz) From the attached Figure 5 、 7 It can be seen that in the low field region there are δ H 7.94, m; δ H 7.93, m; δ H 7.36, d, J = 9.2 Hz; δ H 7.24, m; δ H 7.22, m, a total of 5 benzene ring olefinic hydrogen proton signals, and in the high field region there are δ H 2.63, s; δ H 2.55, s, a total of 2 singlet methyl signals and​δ H 1.34, d, J = 6.9 Hz; δ H 0.84, d, J = 6.8 Hz, two doublet methyl signals in total. Combining with Figure 6 and 8 it can be seen that this compound has 26 carbon signals, among which 6 carbons are of a sugar unit structure, that is δ C / δ H 103.1 / 4.78, 77.5 / 3.49, 78.1 / 3.47, 67.5 / 4.01, 82.0 / 3.70, 68.7 / 3.88 / 3.45. For the remaining 20 carbon signals, combining the characteristics of the number of its methyl groups and olefinic hydrogens, it can be inferred that it is a diterpene structural fragment.

[0032] Furthermore, through the two-dimensional NMR data, the planar structure was analyzed. According to the Figure 9 continuous proton correlation signals in Figure 8 and the lemnaliside A correlation signals, the planar structure of the compound lemnaliside A can be constructed. Compared with the known compounds of this type, the difference of the compound

[0033] is that the decalin ring in its diterpene part becomes naphthalene. lemnaliside A For the determination of the configuration of the compound lemnaliside A , first, by comparing the NMR data with the same type of compounds with single crystal structures and considering from the perspective of biogenetic synthesis, it is determined that the glycoside part of the compound β is R -D-glucose, and the configurations at C-15 and C-16 positions are both S . For the determination of the configuration of the diterpene part, only the configuration at C-11 position of this compound needs to be determined. Considering from the perspective of the biogenetic synthesis pathway, it can be determined that the configuration of C-11 in its diterpene part is 11 Figure 10 . Finally, through the comparison of the TDDFT-ECD calculation shown in lemnaliside A with the experimental CD value, the absolute configuration of the compound

[0034] Example 4 The new diterpene glycoside compound provided in Example 1 of this application lemnaliside A can be used as a candidate molecule for preparing anti-tumor drugs, that is, the new compound lemnaliside A has obvious inhibitory activity against the drug-resistant human small cell lung cancer cell line NCI-H446 / EP. The specific content is as follows: Testing the new diterpene glycoside compound provided in Example 1 by the MTT methodlemnaliside A The inhibitory effect on human chronic myeloid leukemia cell line K562 was also tested by the SRB method for the novel diterpene glycoside compounds provided in Example 1. lemnaliside A The cytotoxic activities against human normal liver cell line L-02, human metastatic pancreatic adenocarcinoma cell line ASPC-1, human breast cancer cell line MDA-MB-231, human small cell lung cancer cell line NCI-H446, and drug-resistant human small cell lung cancer cell line NCI-H446 / EP were tested. Among them, doxorubicin was used as the positive drug, and the test results are shown in Tables 2 and 3 respectively.

[0035] Table 2: Novel diterpene glycoside compounds lemnaliside A Inhibition rate % of the primary screening for anti-tumor activity Table 3: Novel diterpene glycoside compounds lemnaliside A Results of the rescreening for the inhibitory effect on NCI-H446 / EP cells As can be seen from Tables 1 and 2, the novel diterpene glycoside compounds lemnaliside A have a selective inhibitory effect on the drug-resistant human small cell lung cancer cell line NCI-H446 / EP, and its IC 50 value is 17.75 µ μM. This novel diterpene glycoside compound lemnaliside A can be used to prepare drugs for treating drug-resistant human small cell lung cancer.

[0036] The above are only the preferred embodiments of the present application and are not used to limit the present application. For those skilled in the art, various changes and modifications can be made to the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included within the protection scope of the present application.

Claims

1. A new diterpene glycoside compound, characterized in that, The chemical structural formula of the new diterpene glycoside compound is as follows: 。 2. A preparation method of a novel diterpene glycoside compound, characterized in that, Including: S01: The soft coral is cut into pieces and soaked in methanol at room temperature to obtain an extract. After the extract is concentrated under reduced pressure, desalted with anhydrous methanol, and concentrated under reduced pressure again, a crude extract is obtained. S02: After the crude extract is separated by silica gel column chromatography under reduced pressure, eluted with a gradient, and detected by thin-layer chromatography development, the same fractions are combined and concentrated to obtain 16 primary components F1 - F16. Among them, the conditions for silica gel column chromatography under reduced pressure separation are: the chromatography column specifications are 160mm * 500mm, and the silica gel particle size is 200 - 300 mesh; the conditions for thin-layer chromatography development detection are: the developing agent is petroleum ether and propanol with a volume ratio of 3:1, and the developer is a sulfuric acid ethanol solution with a mass concentration of 10%. S03: F16 among the primary components F1 - F16 is separated by silica gel column chromatography, eluted with a gradient, and detected by thin-layer chromatography development. The same fractions are combined and concentrated to obtain 6 secondary components F161 - F166. Among them, the conditions for silica gel column chromatography separation are: the chromatography column specifications are 60mm * 500mm; the silica gel particle size is 300 - 400 mesh; the conditions for thin-layer chromatography development detection are: the developing agent is petroleum ether and propanol with a volume ratio of 3:1, and the developer is a sulfuric acid ethanol solution with a mass concentration of 10%. S04: After the F165 in the secondary components F161 - F166 is analyzed by high performance liquid chromatography, separated by medium pressure preparative liquid chromatography, and isocratically eluted, 4 tertiary components F1651 - F1654 are obtained; among them, the conditions for the high performance liquid chromatography analysis are: the analytical chromatographic column is a Greenherb ODS C column with a size of 4.6 mm * 250 mm and a particle size of 5 μm, and the flow rate is 1 ml / min; the conditions for the medium pressure preparative liquid chromatography separation are: the preparative chromatographic column is a Greenherb ODS C column with a size of 20 mm * 250 mm and a particle size of 5 μm, and the flow rate is 10 ml / min; 18 column, and the flow rate is 1 ml / min; the conditions for the medium pressure preparative liquid chromatography separation are: the preparative chromatographic column is a Greenherb ODS C column with a size of 20 mm * 250 mm and a particle size of 5 μm, 18 column, and the flow rate is 10 ml / min; S05: After the F1654 in the tertiary components F1651 - F1654 is analyzed by high performance liquid chromatography, separated by high performance preparative liquid chromatography, and isocratically eluted, a new diterpene glycoside compound is obtained; among them, the conditions for the high performance liquid chromatography analysis are: the analytical chromatographic column is a Greenherb ODS C column with a size of 4.6 mm * 250 mm and a particle size of 5 μm, and the flow rate is 1 ml / min; the conditions for the high performance preparative liquid chromatography separation are: the chromatographic column is a Greenherb ODS C column with a size of 10 mm * 250 mm and a particle size of 5 μm, and the flow rate is 2 ml / min. 18 column, and the flow rate is 1 ml / min; the conditions for the high performance preparative liquid chromatography separation are: the chromatographic column is a Greenherb ODS C column with a size of 10 mm * 250 mm and a particle size of 5 μm, 18 column, and the flow rate is 2 ml / min.

3. The preparation method of the diterpene glycoside new compound according to claim 2, characterized in that, The soaking in methanol at room temperature includes soaking the cut soft coral in methanol at room temperature 6 times, with each soaking for 3 days; the temperature for concentration under reduced pressure is 37°C and the pressure is 0.1 MPa.

4. The preparation method of the new diterpene glycoside compound according to claim 2, characterized in that, In S02, gradient elution is successively carried out with petroleum ether, petroleum ether / acetone with volume ratios of 100 / 1, 50 / 1, 30 / 1, 20 / 1, 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, dichloromethane, dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, 2 / 1, 1 / 1, 0 / 1.

5. The preparation method of the novel diterpene glycoside compound according to claim 2, characterized in that, In S03, gradient elution is successively carried out with dichloromethane / methanol with volume ratios of 10 / 1, 5 / 1, 3 / 1, 1 / 1.

6. The preparation method of the diterpene glycoside new compound according to claim 2, characterized in that, In S04, isocratic elution is carried out with methanol / water with a volume ratio of 80 / 20.

7. The preparation method of the new diterpene glycoside compound according to claim 2, characterized in that, In S05, isocratic elution is carried out with acetonitrile / water with a volume ratio of 35 / 65.

8. The new diterpene glycoside compound described in claim 1 or the new diterpene glycoside compound prepared by the preparation method described in any one of claims 2 - 7 is used for preparing a drug for treating drug-resistant human small cell lung cancer.

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