New compound with anti-hepatoma activity in dendrobium and its separation and preparation method and application

Abstract

The application discloses a new compound with anti-hepatoma activity in dendrobium, and a separation and preparation method and application thereof, and belongs to the technical field of natural medicines. The preparation method comprises the following steps: step one, taking dried stems of dendrobium, crushing, and extracting at least two times by heating reflux with 95% ethanol for at least two hours each time, combining the extract after filtration, and concentrating into thick extract under reduced pressure to obtain a crude extract; step two, dispersing the crude extract with water, and sequentially extracting with petroleum ether and dichloromethane; concentrating the water phase under reduced pressure after extraction to obtain concentrated water liquid; step three, loading the concentrated water liquid on a macroporous resin column (filler is AB-8 or D101), and gradient eluting with an ethanol-water solvent system; and step four, isocratic eluting with 35% acetonitrile-water solvent by preparative liquid phase. The new sesquiterpene glycoside compound is separated from traditional Chinese medicine dendrobium, the new compound has good anti-hepatoma cell activity, and lays a foundation for development and utilization of dendrobium plant resources and new drug research and development.

Description

Technical Field

[0001] This invention relates to novel compounds isolated from the traditional Chinese medicine Dendrobium, their preparation methods and applications, specifically to a class of novel compounds in Dendrobium with anti-hepatocellular carcinoma activity, their isolation and preparation methods and applications, belonging to the field of natural medicine technology. Background Technology

[0002] Dendrobium nobile (scientific name: Dendrobium nobile) Dendrobium nobile Lindl. is a member of the Orchidaceae family ( Orchidaceae Dendrobium ( ) Dendrobium Dendrobium nobile is a perennial epiphytic herb. It is a traditional and precious Chinese medicinal herb, included in the Chinese Pharmacopoeia. Its dried stems have the effects of nourishing the stomach and promoting the production of body fluids, nourishing yin and clearing heat. Modern research shows that it also has various pharmacological effects such as enhancing immunity, anti-tumor activity, and anti-aging.

[0003] Existing technologies disclose that *Dendrobium nobile* mainly contains compounds such as dendrobine, dendrobine ether, dendrobamine, carotene, and methylene dendrobine. Although *Dendrobium nobile* is a traditional and precious medicinal herb, compared to bulk medicinal herbs such as ginseng and astragalus, its modern systematic research on chemical components lags significantly behind. An incomplete compound library is a real bottleneck restricting its in-depth development. Currently, research on the chemical components of *Dendrobium nobile* largely focuses on components with clearly defined pharmacological activities, with insufficient systematic exploration of structurally complex and functionally unknown trace components (such as special sesquiterpenes). Therefore, there is an urgent need for a new compound isolated from *Dendrobium nobile*, along with its preparation method and applications. Summary of the Invention

[0004] To address the aforementioned problems, the purpose of this invention is to provide a new compound isolated from Dendrobium nobile, specifically a new compound from Dendrobium nobile with anti-hepatocellular carcinoma activity, which is a novel sesquiterpene glycoside natural drug.

[0005] Meanwhile, this invention provides a method for the isolation and preparation of novel compounds from Dendrobium nobile.

[0006] Meanwhile, this invention provides the application of a novel compound isolated from Dendrobium nobile in the preparation of drugs for treating liver cancer.

[0007] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows: A new class of compounds from Dendrobium with anti-liver cancer activity has been identified. The structure of compound (I) is as follows: ; The structure of compound (II) is as follows: .

[0008] A method for isolating and preparing a new class of compounds with anti-hepatocellular carcinoma activity from Dendrobium, comprising the following steps: Step 1: Take Dendrobium nobile Dendrobium nobile Lindl. dried aerial stems, crushed, sieved, and extracted with 95% ethanol under reflux at least twice, each time using 12-15 times the amount of raw material, for at least 2 hours each time. After filtration, the extracts were combined and concentrated under reduced pressure to obtain a thick extract. Step 2: After dispersing the crude extract in water, it is extracted sequentially with petroleum ether and dichloromethane. The aqueous phase after extraction is then concentrated under reduced pressure to obtain a concentrated aqueous solution. Step 3: Load the concentrated aqueous solution onto a macroporous resin column (packing material is AB-8 or D101) and elute with a gradient elution system of ethanol-water solvent. Step 4: After concentrating the 30% ethanol eluent, the solution is then subjected to preparative liquid chromatography with 35% acetonitrile-water solution as the mobile phase and a detection wavelength of 210 nm. The chromatographic peak with a retention time of 21.5 min is collected to obtain the crude compound (I); the chromatographic peak with a retention time of 28.0 min is collected to obtain the crude compound (II). Step 5: The crude compounds (I) and (II) were purified by Sephadex LH-20 gel column chromatography. Compound (I) was obtained by elution with methanol-chloroform (1:1), and compound (II) was obtained by elution with methanol.

[0009] Preferably, in step one, the powder is passed through a 40-60 mesh sieve.

[0010] Preferably, in step two, the boiling range of petroleum ether is 60-90℃.

[0011] Preferably, in step two, the amount of water used is 10-14 times the weight of the crude extract; the amount of petroleum ether and dichloromethane used in each extraction is the same volume as water, and each extractant is extracted at least 3 times.

[0012] Preferably, in step three, the ethanol-water solvent system uses water, 30%, 60%, 80% and 100% EtOH / H2O as eluents in sequence, eluting 3-4 column volumes for each gradient; the diameter-to-height ratio of the macroporous resin column is 1:(6-10).

[0013] Preferably, in step four, the reversed-phase C18 column used for preparing the liquid phase is an Agela Venusil PrepG C18 column, the injection volume is 60-120 mg, and the flow rate is 15 mL / min.

[0014] Preferably, in step five, the diameter-to-height ratio of the Sephadex LH-20 gel column is 1:(30-50). After loading the sample, the column is eluted with methanol-chloroform (1:1) for at least 5 column volumes. The eluent is collected, concentrated, and dried to obtain compound (I). The column is then eluted with methanol for at least 5 column volumes. The eluent is collected, concentrated, and dried to obtain compound (II).

[0015] The application of a novel compound from Dendrobium with anti-hepatocellular carcinoma activity in the preparation of anti-hepatocellular carcinoma drugs.

[0016] Specifically, the use of compound (I) in the preparation of anti-hepatocellular carcinoma drugs.

[0017] Specifically, the application of compound (II) in the preparation of anti-liver cancer drugs.

[0018] An anti-liver cancer drug is prepared using a novel compound from Dendrobium that has anti-liver cancer activity.

[0019] Specifically, an anti-liver cancer drug is prepared using compound (I) of the present invention.

[0020] Specifically, an anti-liver cancer drug is prepared using compound (II) of the present invention.

[0021] Compared with the prior art, the present invention has the following advantages: This invention isolates two novel sesquiterpene glycosides from *Dendrobium nobile*. These sesquiterpene glycosides are unique sesquiterpenes found in *Dendrobium nobile*, characterized by low content, complex structures, and significant isolation challenges. These new compounds exhibit excellent anti-hepatocellular carcinoma activity. Hepatocellular carcinoma is one of the most common malignant solid tumors, and the search for highly effective and low-toxicity natural anti-tumor lead compounds has always been a hot topic in drug development. This invention lays the foundation for the development and utilization of *Dendrobium nobile*, enriches its chemical composition library, and provides important lead compounds for the development of new anti-hepatocellular carcinoma drugs. Attached Figure Description

[0022] Figure 1 For the compound (I) of the present invention 1 H-NMR spectrum; Figure 2 For the compound (I) of the present invention 13 C-NMR spectrum; Figure 3 The DEPT spectrum of compound (I) of this invention; Figure 4 The HSQC spectrum of compound (I) of this invention; Figure 5 The HMBC spectrum of compound (I) of this invention; Figure 6 The HH COSY spectrum of compound (I) of this invention; Figure 7 The NOESY spectrum of compound (I) of this invention; Figure 8 For the compound (II) of the present invention 1 H-NMR spectrum; Figure 9 For the compound (II) of the present invention13 C-NMR spectrum; Figure 10 The DEPT spectrum of compound (II) of this invention; Figure 11 The HSQC spectrum of compound (Ⅱ) of this invention; Figure 12 The HMBC spectrum of compound (II) of this invention; Figure 13 The HH COSY spectrum of compound (II) of this invention; Figure 14 This is the NOESY spectrum of compound (Ⅱ) of the present invention. Detailed Implementation

[0023] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0024] Source of Dendrobium nobile: The Dendrobium nobile sample used in the experiment was collected in September 2023 from Tiantai Town, Chishui City, Guizhou Province (longitude: 105.755599, latitude: 28.562458, altitude: 825 meters). The specimen was identified as Dendrobium nobile of the Orchidaceae family by Professor Zhao Jiancheng of the School of Life Sciences, Hebei Normal University. Dendrobium nobile Dried stem of Lindl. Voucher specimen (20230901) is deposited in the Department of Processing of Traditional Chinese Medicine, Hebei University of Traditional Chinese Medicine. Example 1

[0025] A new compound (I) from Dendrobium has anti-liver cancer activity, and its structure is as follows: ; The structure of compound (II) is as follows: .

[0026] A method for isolating and preparing a new class of compounds with anti-hepatocellular carcinoma activity from Dendrobium, comprising the following steps: Step 1: Take Dendrobium nobile Dendrobium nobile Lindl. dried aerial stems, crushed and passed through a 40-mesh sieve, extracted twice with 95% ethanol under reflux, each time using 13 times the amount of raw material, each extraction lasting 2 hours, the extracts were combined after filtration and concentrated under reduced pressure to obtain a thick extract, thus obtaining the crude extract.

[0027] Step 2: After dispersing the crude extract in water, extract it sequentially with petroleum ether (boiling range 60-90℃) and dichloromethane. After extraction, concentrate the aqueous phase under reduced pressure to obtain concentrated aqueous solution. The amount of water used is 13 times the weight of the crude extract. The amount of petroleum ether and dichloromethane used in each extraction is the same volume as water, and each extractant is extracted 3 times.

[0028] Step 3: Load the concentrated aqueous solution onto a macroporous resin column (packing material is AB-8) and elute using an ethanol-water solvent system in a gradient. The ethanol-water solvent system uses water, 30%, 60%, 80%, and 100% EtOH / H2O as eluents in sequence, with each gradient eluting for 3 column volumes. The diameter-to-height ratio of the macroporous resin column is 1:8.

[0029] Step 4: Collect the 30% ethanol eluent. After concentrating the 30% ethanol eluent, perform preparative liquid chromatography (PCC) with a mobile phase of 35% acetonitrile-water solution and a detection wavelength of 210 nm. Collect the chromatographic peak with a retention time of 21.5 min to obtain crude compound (I); collect the chromatographic peak with a retention time of 28.0 min to obtain crude compound (II). The reversed-phase C18 column used in the PCC was an Agela Venusil PrepG C18 preparative column, with an injection volume of 100 mg and a flow rate of 15 mL / min. Step 5: Purify the crude compounds (I and II) using Sephadex LH-20 gel columns. The Sephadex LH-20 column has a diameter-to-height ratio of 1:40. After loading the sample, elute with methanol-chloroform (1:1) for 5 column volumes, collect the eluent, concentrate and dry to obtain compound (I); elute with methanol for 5 column volumes, collect the eluent, concentrate and dry to obtain compound (II).

[0030] The application of the novel compound in the preparation of an anti-liver cancer drug in this embodiment.

[0031] An anti-liver cancer drug was prepared using the novel compound described in this embodiment.

[0032] Example 2

[0033] The only difference between this embodiment and Embodiment 1 is that: A method for isolating and preparing a new class of compounds with anti-hepatocellular carcinoma activity from Dendrobium, comprising the following steps: Step 1: Take Dendrobium nobile Dendrobium nobile Lindl. dried aerial stems, crushed and passed through a 60-mesh sieve, extracted three times by heating and reflux with 95% ethanol, each time using a solvent amount equivalent to 12 times the amount of raw material, each time for 2 hours, after filtration, the extracts were combined and concentrated under reduced pressure to obtain a thick extract, thus obtaining the crude extract.

[0034] Step 2: After dispersing the crude extract in water, extract it sequentially with petroleum ether and dichloromethane. After extraction, concentrate the aqueous phase under reduced pressure to obtain concentrated aqueous solution. The amount of water used is 10 times the weight of the crude extract. The amount of petroleum ether and dichloromethane used in each extraction is the same volume as water, and each extractant is extracted 4 times.

[0035] Step 3: Load the concentrated aqueous solution onto a macroporous resin column (packing material is D101) and elute using an ethanol-water solvent system in a gradient. The ethanol-water solvent system uses water, 30%, 60%, 80%, and 100% EtOH / H2O as eluents in sequence, with each gradient eluting for 4 column volumes. The diameter-to-height ratio of the macroporous resin column is 1:6.

[0036] Step 4: Collect the 30% ethanol eluent. After concentrating the 30% ethanol eluent, perform preparative liquid chromatography using 35% acetonitrile-water solution as the mobile phase and a detection wavelength of 210 nm. Collect the chromatographic peak with a retention time of 21.5 min to obtain the crude compound (I); collect the chromatographic peak with a retention time of 28.0 min to obtain the crude compound (II). The reversed-phase C18 column used in the preparative liquid chromatography was an Agela Venusil PrepG C18 preparative column, with an injection volume of 60 mg and a flow rate of 15 mL / min.

[0037] Step 5: The crude compounds (I) and (II) were purified by Sephadex LH-20 gel column purification. The diameter-to-height ratio of the Sephadex LH-20 gel column was 1:30. After loading the sample, 6 column volumes of methanol-chloroform (1:1) were eluted. The eluent was collected, concentrated, and dried to obtain compound (I). 6 column volumes of methanol were eluted. The eluent was collected, concentrated, and dried to obtain compound (II).

[0038] Example 3

[0039] The only difference between this embodiment and Embodiment 1 is that: A method for isolating and preparing a new class of compounds with anti-hepatocellular carcinoma activity from Dendrobium, comprising the following steps: Step 1: Take Dendrobium nobile Dendrobium nobile Lindl. dried aerial stems, crushed and passed through a 50-mesh sieve, extracted four times with 95% ethanol under reflux, each time using 12 times the amount of solvent as feed, for 3 hours each time. After filtration, the extracts were combined and concentrated under reduced pressure to obtain a thick extract, thus obtaining the crude extract.

[0040] Step 2: After dispersing the crude extract with hot water, it is extracted sequentially with petroleum ether and dichloromethane. The aqueous phase is then concentrated under reduced pressure to obtain a concentrate. The amount of hot water used is 14 times the weight of the crude extract. The amount of petroleum ether and dichloromethane used in each extraction is the same volume as the hot water. Each extractant is extracted 3 times. The temperature of the hot water is 90℃.

[0041] Step 3: Load the concentrated aqueous solution onto a macroporous resin column (packing material is D101) and elute using an ethanol-water solvent system in a gradient. The ethanol-water solvent system uses water, 20%, 50%, 70%, and 100% EtOH / H2O as eluents in sequence, with each gradient eluting for 4 column volumes. The diameter-to-height ratio of the macroporous resin column is 1:10.

[0042] Step 4: Collect the 30% ethanol eluent. After concentrating the 30% ethanol eluent, perform preparative liquid chromatography using 35% acetonitrile-water solution as the mobile phase and a detection wavelength of 210 nm. Collect the chromatographic peak with a retention time of 21.5 min to obtain the crude compound (I); collect the chromatographic peak with a retention time of 28.0 min to obtain the crude compound (II). The reversed-phase C18 column used in the preparative liquid chromatography was an Agela Venusil PrepG C18 preparative column, with an injection volume of 120 mg and a flow rate of 15 mL / min.

[0043] Step 5: The crude compounds (I) and (II) were purified by Sephadex LH-20 gel column purification. The diameter-to-height ratio of the Sephadex LH-20 gel column was 1:50. After loading the sample, 5 column volumes of methanol-chloroform (1:1) were eluted. The eluent was collected, concentrated, and dried to obtain compound (I). 5 column volumes of methanol were eluted. The eluent was collected, concentrated, and dried to obtain compound (II).

[0044] The English name of compound (I) is: 2,4,13-Trihydroxy-9-hydroxymethyl-1,12-dimethyl-3-oxatricyclo[1.6]undecan-15-one-11-O- β -D-glucopyranoside.

[0045] Compound (I): Off-white solid; maximum ultraviolet absorption wavelength λmax = 194 nm; =-8.86° (C=0.079, MeOH).

[0046] .

[0047] Identification of the structure of compound (I): such as Figures 1-7 As shown, the NMR data of compound (I) are shown in Table 1 below.

[0048] Table 1. NMR data (600 MHz, C5D5N) for compound (I)

[0049] Compound (I) HR-ESIMS m / z The display shows [M+NH4]. + Ion signal 480.2479 (theoretical value: 462.4881). 13 C1-NMR data showed 21 carbon atoms, which together led to the deduction that the molecular formula is C10. 21 H 34 O11 The degree of unsaturation is 5. 1 H-NMR, 13 C-NMR and DEPT135 signals show [ δ H 1.14 (3H, s, H-10), 1.26 (3H, d, J =6.7Hz, H-14)] is 2 methyl groups, [ δ C [48.5 (C-1), 80.8 (C-4)] represents two quaternary carbons, [ δ C 179.7 (C-15)] is the ester carbonyl signal. H-5 and H-6, H-6 and H-7, H-7 and H-8, H-8 and H-9, H-9 and H-1 1 H- 1 The HMBC correlations between H-2 and C-4, H-3 and C-1, C-2, H-9 and C-1, C-8, and H-10 and C-1, C-6 collectively confirm that the parent ring structure of this compound is a six-membered ring coupled to a five-membered ring. The HMBC relationships between H-3 and C-15, and H-5 and C-15 confirm that the C-3 hydroxyl group forms a lactone bond with the C-5 carboxyl group. The HMBC correlation signals between H-1' and C-3', H-2' and C-4', H-4' and C-3', C-5', and H-6' and C-5', along with the relatively large coupling constant of 7.7 Hz for H-1', collectively indicate that... β -D-glucose, H-1' and C-11, H-11 and C-1' HMBC related signals are shown. β -D-glucose C-1'-OH is attached at C-11. HMBC-related signals of H-13 with C-4 and C-12, and H-14 with C-12 confirm the connection mode of the C-4 substituent. Compound (I) is named 2,4,13-Trihydroxy-9-hydroxymethyl-1,12-dimethyl-3-oxatricyclo[1.6]undecan-15-one-11-O- β -D-glucopyranoside.

[0050] The English name of compound (II) is: 5 β, 8 β, 12-trihydroxy-cyclopacamphane-12-O- β -D-glucopyranoside.

[0051] Compound (II): White solid; maximum ultraviolet absorption wavelength λmax = 194 nm; =+15.76° (C=0.165, MeOH).

[0052]

[0053] Structural identification of compound (II): such as Figures 8-14 As shown, the NMR data of compound (II) are shown in Table 2 below.

[0054] Table 2 NMR data of compound (II) (600MHz, C5D5N)

[0055] Compound (II) HRESIMS (m / z) shows [M+NH4] + Ion signal 432.2605 (theoretical value: 414.4950). 13 C-NMR data showed that 21 carbon atoms together led to the molecular formula being C. 21 H 34 O8 has an unsaturation degree of 5. 1 H-NMR, 13 C-NMR and DEPT135 signals show [ δ H 1.96 (3H, s, H-14), 1.49 (3H, s, H-15), 1.10 (3H, d, J =6.7Hz,H-13)] is 3 methyl groups, [ δ C 24.1 (C-3), 49.9 (C-7)] represent two quaternary carbon signals. H-2 and H-4, H-4 and H-5, H-5 and H-6, H-8 and H-9, H-9 and H-10, H-10 and H-11... 1 H- 1 H COSY correlation, H-1 with C-1, C-3, H-4 with C-6, H-9 with C-7, C-8, C-10, H-13 with C-11, C-12, H-14 with C-7, H-15 with C-3, C-4 HMBC correlation, and H-1 with H-2, H-1 with H-6, H-2 with H-4 NOEY correlation. H-1' with H-2', H-3' with H-4, and 'H-4' with H-5' HMBC correlation. 1 H- 1 The correlation between H-4' and C-3', C-5', the NOESY correlation between H-1' and H-3', and H-1' and H-5', along with the large coupling constant of 7.7Hz of H-1', collectively confirm that... β -D-glucose, H-1' and C-12, H-12 and C-1' HMBC related signals are shown.β -D-glucose C-1'-OH is attached at the C-12 position. Compound (II) is named 5. β, 8 β, 12-trihydroxy-cyclopacamphane-12-O- β -D-glucopyranoside.

[0056] Pharmacological activity: Human hepatocellular carcinoma cell lines HepG2, Hep3B, and HuH-7 were cultured in high-glucose DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. All cells were cultured in a 37°C, 5% CO2 incubator. The culture medium was replaced every 1-2 days. When cell confluence reached 80%-90%, cells were trypsinized and passaged at a ratio of 1:3-1:4, approximately every 3-4 days. All experiments were performed when cells were in good condition and in the logarithmic growth phase.

[0057] MTT assay was used to determine cell proliferation. HepG2, Hep3B, and HuH-7 cells in logarithmic growth phase were injected at a dose of 1×10⁻⁶. 5 Cells were seeded at 100 μL / well in 96-well plates, with 100 μL of cell suspension added to each well. The plates were incubated at 37°C with 5% CO2 for 24 h until complete cell adhesion. Drug treatment was then initiated. Three control groups were established: a blank control group (containing only complete culture medium, no cells), a negative control group (containing cells and complete culture medium without drug), and drug treatment groups (containing cells and culture medium with different drug concentration gradients). Each group had six replicates. The original culture medium in the 96-well plates was discarded. 100 μL of drug-free complete culture medium was added to each well of the blank and negative control groups. 100 μL of culture medium containing different concentrations (100, 50, 25, 12.5, 6.25 μmol / L) of drug was added to each well of the drug treatment groups. The plates were gently shaken to mix the drug, and the plates were incubated for another 48 h. Then, 10 μL of MTT (5 mg / mL) solution was added to each well, and the plates were incubated in the dark for 4 h. Carefully aspirate the supernatant, add 150 μL of DMSO to each well, shake in the dark for 10 min, and measure the absorbance (A) of each well at 570 nm using a microplate reader. Calculate the cell viability using the formula.

[0058] Cell viability (%) = (Mean A value of drug-treated groups - Mean A value of blank control group) / (Mean A value of negative control group - Mean A value of blank control group) × 100% Table 3. Cytotoxicity of the new compounds

[0059] Pharmacological activity results showed that compounds (I) and (II) exhibited varying degrees of inhibitory activity against the proliferation of HepG2, Hep3B, and HuH-7 cells. Specifically, compound (I) showed the greatest inhibitory effect on the IC50 of HepG2 cells. 50 The concentration was (7.58 ± 0.85) μmol / L, which was not significantly different from the positive control cisplatin (6.75 ± 0.42) μmol / L, indicating comparable activity; compound (II) showed an IC50 response in Hep3B cells. 50 The concentration was (5.13 ± 0.47) μmol / L, which was not significantly different from that of cisplatin (4.92 ± 0.56) μmol / L, indicating comparable activity.

[0060] It should be understood that, in order to simplify this disclosure and aid in understanding one or more of the various aspects of the invention, features of the invention are sometimes grouped together in a single embodiment or in its description in the foregoing description of exemplary embodiments of the invention. However, this method of disclosure should not be construed as reflecting an intention that the claimed invention requires more features than expressly recited in each claim. Rather, as reflected in the claims, inventive aspects lie in fewer than all the features of the foregoingly disclosed embodiments. Therefore, the claims, following the detailed description, are hereby expressly incorporated into that detailed description, wherein each claim itself is a separate embodiment of the invention.

[0061] Although the invention has been described with reference to a limited number of embodiments, those skilled in the art will understand from the foregoing description that other embodiments are conceivable within the scope of the invention described herein. Furthermore, it should be noted that the language used in this specification has been chosen primarily for readability and instructional purposes, and not for the purpose of interpreting or limiting the subject matter of the invention. Therefore, many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the appended claims. The disclosure of the invention is illustrative and not restrictive, and the scope of the invention is defined by the appended claims.

[0062] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A new class of compounds from Dendrobium with anti-hepatocellular carcinoma activity, characterized in that, Its structure is as follows: ； 。 2. The method for isolating and preparing a new type of compound with anti-hepatocellular carcinoma activity from Dendrobium according to claim 1, characterized in that, Includes the following steps: Step 1, take Dendrobium Dendrobium nobile Lindl. dried aerial stems, crushed, sieved, and extracted with 95% ethanol under reflux at least twice, each time using 12-15 times the amount of raw material, for at least 2 hours each time. After filtration, the extracts were combined and concentrated under reduced pressure to obtain a thick extract. Step 2: After dispersing the crude extract in water, it is extracted sequentially with petroleum ether and dichloromethane. The aqueous phase after extraction is then concentrated under reduced pressure to obtain a concentrated aqueous solution. Step 3: Load the concentrated aqueous solution onto a macroporous resin column and elute with a gradient using an ethanol-water solvent system; Step 4: After concentrating the 30% ethanol eluent, the solution is then subjected to preparative liquid chromatography with 35% acetonitrile-water solution as the mobile phase and a detection wavelength of 210 nm. The chromatographic peak with a retention time of 21.5 min is collected to obtain the crude compound (I). The chromatographic peak with a retention time of 28.0 min was collected to obtain crude compound (II); Step 5: The crude compounds (I) and (II) were purified by Sephadex LH-20 gel column chromatography. Compound (I) was obtained by elution with methanol-chloroform (1:1), and compound (II) was obtained by elution with methanol.

3. The separation and preparation method according to claim 2, characterized in that, In step one, the powder is pulverized and then passed through a 40-60 mesh sieve.

4. The separation and preparation method according to claim 2, characterized in that, In step two, the boiling range of petroleum ether is 60-90℃.

5. The separation and preparation method according to claim 2, characterized in that, In step two, the amount of water used is 10-14 times the weight of the crude extract; the amount of petroleum ether and dichloromethane used in each extraction is the same volume as the water, and each extractant is extracted at least 3 times.

6. The separation and preparation method according to claim 2, characterized in that, In step three, the ethanol-water solvent system uses water, 30%, 60%, 80%, and 100% EtOH / H2O as eluents in sequence, eluting 3-4 column volumes for each gradient; the diameter-to-height ratio of the macroporous resin column is 1:(6-10); the macroporous resin column packing is AB-8 or D101.

7. The separation and preparation method according to claim 2, characterized in that, In step four, the reversed-phase C18 column used for preparing the liquid phase is an Agela Venusil PrepG C18 column, with an injection volume of 60-120 mg and a flow rate of 15 mL / min.

8. The separation and preparation method according to claim 2, characterized in that, In step five, the diameter-to-height ratio of the Sephadex LH-20 gel column is 1:(30-50). After loading the sample, elute with methanol-chloroform (1:1) for at least 5 column volumes, collect the eluent, concentrate and dry to obtain compound (I); elute with methanol for at least 5 column volumes, collect the eluent, concentrate and dry to obtain compound (II).

9. The application of a novel compound from Dendrobium with anti-hepatocellular carcinoma activity as described in claim 1 in the preparation of anti-hepatocellular carcinoma drugs.

10. An anti-liver cancer drug, characterized in that, It was prepared using a novel compound from Dendrobium with anti-liver cancer activity as described in claim 1.

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