Glucoside compound as well as preparation method and application thereof

Abstract

The invention provides a glycoside compound as well as a preparation method and application thereof, and relates to the technical field of natural medicine extraction. The glycoside compound provided by the invention has the following structure, the glycoside compound disclosed by the invention is extracted from a phyllostachys plant; the glycoside compound can be used for preparing medicines for treating diabetes, medicines for treating diabetic complications or medicines for treating neurodegenerative diseases. Furthermore, the glycoside compound shown in the formula I participates in the processes of energy metabolism, inflammatory response, immune cell functions, cell growth and survival, response of cells to hypoxia and the like, and has a treatment effect on diabetes and complications thereof; the glycoside compound in the formula II mainly acts on a mitochondrial respiratory chain complex I, regulates mitochondrial functions and energy metabolism, and has a treatment effect on neurodegenerative diseases; the glycoside compound shown in the formula III and the glycoside compound shown in the formula IV focus on regulation of cell metabolism and signal transduction and have a treatment effect on nervous system diseases. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the technical field of natural drug extraction, and specifically relates to a glycoside compound and a preparation method and application thereof. Background Art

[0002] Bamboo has a long history of medicinal use in my country, and bamboo stalks, leaves and roots can all be used as medicine. Phyllostachys is the genus with the greatest economic value in the bamboo subfamily, with 70 species discovered. Among them, there are 11 species of medicinal bamboo, including P. reticulata, P. edulis, P. glauca, P. heteroclada, P. makinoi, P. nidularia, P. nigra, P. nigra var. henonis, P. sulphurea, P. violascens and P. sulphurea f. robertii, which have good therapeutic effects on symptoms such as inflammation, fever and cough.

[0003] The golden bamboo is a variant of purple bamboo, with a culm height of about 7 to 18 meters, a culm wall thickness of up to 5 mm, and very few dark brown spots on the top of the sheath. This bamboo species is widely distributed in provinces south of the Yangtze River Basin. It is one of the medicinal bamboo species. The culm part can be used as medicine, and the culm of the golden bamboo is the raw material for preparing the traditional Chinese medicine bamboo juice and bamboo shavings, which have the effects of expectoration and cough relief.

[0004] Modern pharmacological studies have shown that bamboo extracts have anti-inflammatory, anti-cancer, cholesterol-lowering, melanin-inhibiting and neuroprotective effects. The rich secondary metabolites in bamboo provide the material basis for its diverse pharmacological activities. However, there are relatively few studies on bamboo secondary metabolites, which cannot meet the requirements of the development of modern Chinese medicine. Therefore, the separation and identification of secondary metabolites of Phyllostachys plants is of great value for discovering new active ingredients or lead compounds. Summary of the invention

[0005] The object of the present invention is to provide a glycoside compound and a preparation method and application thereof. The present invention extracts a glycoside compound from a plant of the genus Phyllostachys, and the glycoside compound can be used to prepare a drug for treating diabetes, a drug for treating diabetic complications or a drug for neurodegenerative diseases.

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

[0007] A glycoside compound having the following structure:

[0008]

[0009] The present invention also provides a method for preparing the glycoside compound described in the above technical solution, comprising the following steps:

[0010] The plant of Phyllostachys genus is mixed with ethanol, extracted and concentrated to obtain an alcohol extract;

[0011] Dissolving and resuspending the alcohol extract in distilled water to obtain a suspension; mixing the suspension with butanol, extracting and concentrating the mixture to obtain a butanol phase extract;

[0012] The butanol phase extract is subjected to adsorption resin column separation, and after concentration, a 20% ethanol elution component and a 40% ethanol elution component are obtained; the eluents used in the adsorption resin column separation are water, 20% ethanol and 40% ethanol, respectively;

[0013] The 20% ethanol elution component is separated by a first chromatographic column, and the same fractions are combined and concentrated to obtain components A1 to A10; the eluents used for the first chromatographic column separation are 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 50% ethanol and 100% ethanol, respectively;

[0014] The A3 component is subjected to a first gel column chromatography separation, and the same fractions are combined and concentrated to obtain components A3a to A3s; the mobile phase of the first gel column chromatography separation is water;

[0015] The A3c component is separated by a second chromatographic column to obtain the glycoside compound of formula I, and the eluent used for the second chromatographic column separation is 20% methanol; the A3e component is separated by a third chromatographic column to obtain the glycoside compound of formula II, and the eluent used for the third chromatographic column separation is 15% methanol;

[0016] The 40% ethanol elution group is subjected to a fourth chromatographic column separation, and the same fractions are combined and concentrated to obtain components B1 to B9; the eluents used for the fourth chromatographic column separation are 20% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 60% ethanol, 80% ethanol and 100% ethanol, respectively;

[0017] The B1 component is subjected to a second gel column chromatography separation, and after concentration, components B1a to B1o are obtained; the mobile phase of the second gel column chromatography separation is water;

[0018] The B1a component is sequentially separated by a fifth chromatographic column to obtain the glycoside compound of formula III and the glycoside compound of formula IV. The eluent used in the fifth chromatographic column separation is 20% methanol.

[0019] Preferably, the extraction comprises: extracting with 90-100% ethanol for 1-2 times and with 70-80% ethanol for 2-3 times in sequence, each extraction lasting 20-30 hours; and the solid-liquid ratio of the Phyllostachys plant to ethanol is 1:3-5 kg / L.

[0020] Preferably, the extraction is performed 3 to 5 times; the butanol is water-saturated n-butanol; and the volume ratio of the butanol to the suspension is 1:1 to 3.

[0021] Preferably, the adsorption resin in the adsorption resin column separation is HP-20.

[0022] Preferably, the first chromatographic column separation and the fourth chromatographic column separation are independently ODS chromatographic column separations, and the filler is ODS-A.

[0023] Preferably, the fillers of the first gel column chromatography separation and the second gel column chromatography separation are independently Sephadex LH-20.

[0024] Preferably, the second chromatographic column separation, the third chromatographic column separation and the fifth chromatographic column separation are independently ODS chromatographic column separations, and the filler is ODS-AQ.

[0025] The present invention also provides the use of the glycoside compound described in the above technical solution in the preparation of a drug for treating diabetes, a drug for treating diabetic complications or a drug for neurodegenerative diseases.

[0026] The present invention provides a glycoside compound having the following structure; the glycoside compound of the present invention is extracted from a plant of the genus Phyllostachys; the glycoside compound can be used to prepare a drug for treating diabetes, a drug for treating diabetic complications, or a drug for treating neurodegenerative diseases. Further, the glycoside compound of formula I of the present invention mainly participates in processes such as energy metabolism, inflammatory response, immune cell function, cell growth and survival, and cell response to hypoxia, and has a therapeutic effect on diabetes and its complications; the glycoside compound of formula II mainly acts on mitochondrial respiratory chain complex I, regulates mitochondrial function and energy metabolism, and has a therapeutic effect on neurodegenerative diseases; the glycoside compound of formula III and the glycoside compound of formula IV focus on regulating cell metabolism and signal transduction, and also have a therapeutic effect on nervous system diseases.

[0027] BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0029] Figure 1 The predicted target PPIs map, key subnetwork map and key target map of the glycoside compound of formula I;

[0030] Figure 2 GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula I;

[0031] Figure 3 The predicted target PPIs map, key subnetwork map and key target map of the glycoside compound of formula II;

[0032] Figure 4 The GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula II;

[0033] Figure 5 The predicted target PPIs map, key subnetwork map and key target map of the glycoside compound of formula III;

[0034] Figure 6 The GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula III;

[0035] Figure 7 The predicted target PPIs map, key subnetwork map and key target map of the glycoside compound of formula IV;

[0036] Figure 8 GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula IV. DETAILED DESCRIPTION

[0037] The present invention provides a glycoside compound having the following structure:

[0038]

[0039] The present invention also provides a method for preparing the glycoside compound described in the above technical solution, comprising the following steps:

[0040] The plant of Phyllostachys genus is mixed with ethanol, extracted and concentrated to obtain an alcohol extract;

[0041] Dissolving and resuspending the alcohol extract in distilled water to obtain a suspension; mixing the suspension with butanol, extracting and concentrating the mixture to obtain a butanol phase extract;

[0042] The butanol phase extract is subjected to adsorption resin column separation, and after concentration, a 20% ethanol elution component and a 40% ethanol elution component are obtained; the eluents used in the adsorption resin column separation are water, 20% ethanol and 40% ethanol, respectively;

[0043] The 20% ethanol elution component is separated by a first chromatographic column, and the same fractions are combined and concentrated to obtain components A1 to A10; the eluents used for the first chromatographic column separation are 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 50% ethanol and 100% ethanol, respectively;

[0044] The A3 component is subjected to a first gel column chromatography separation, and the same fractions are combined and concentrated to obtain components A3a to A3s; the mobile phase of the first gel column chromatography separation is water;

[0045] The A3c component is separated by a second chromatographic column to obtain the glycoside compound of formula I, and the eluent used for the second chromatographic column separation is 20% methanol; the A3e component is separated by a third chromatographic column to obtain the glycoside compound of formula II, and the eluent used for the third chromatographic column separation is 15% methanol;

[0046] The 40% ethanol elution group is subjected to a fourth chromatographic column separation, and the same fractions are combined and concentrated to obtain components B1 to B9; the eluents used for the fourth chromatographic column separation are 20% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 60% ethanol, 80% ethanol and 100% ethanol, respectively;

[0047] The B1 component is subjected to a second gel column chromatography separation, and after concentration, components B1a to B1o are obtained; the mobile phase of the second gel column chromatography separation is water;

[0048] The B1a component is sequentially separated by a fifth chromatographic column to obtain the glycoside compound of formula III and the glycoside compound of formula IV. The eluent used in the fifth chromatographic column separation is 20% methanol.

[0049] The present invention mixes a plant of the genus Phyllostachys with ethanol, extracts and concentrates it to obtain an alcohol extract. In the present invention, the plant of the genus Phyllostachys preferably includes one of Phyllostachys pubescens, Phyllostachys laurel, Phyllostachys pubescens, Phyllostachys pubescens, Phyllostachys edulis, Phyllostachys truncatum, Phyllostachys truncatum, Phyllostachys edulis, Phyllostachys truncatum, Phyllostachys purpurogenum, Phyllostachys praecox, and Phyllostachys leucophylla, more preferably Phyllostachys purpurogenum, and the purple bamboo is more preferably Phyllostachys pubescens; the Phyllostachys pubescens is preferably one or more of the bamboo stalks, bamboo leaves and bamboo whips of Phyllostachys pubescens, more preferably bamboo stalks; the particle size of the plant of the genus Phyllostachys genus is preferably 20 to 60 meshes, more preferably 20 to 40 meshes; if the particle size of the plant of the genus Phyllostachys genus does not meet the above range, it is also preferably crushed; the present invention has no special limitation on the crushing, as long as the desired particle size is reached. In the present invention, the solid-liquid ratio of the plant of the genus Phyllostachys genus to ethanol is preferably 1:3 to 5 kg / L, more preferably 1:3 to 4 kg / L.

[0050] In the present invention, the extraction preferably includes: extracting with 90-100% ethanol 1-2 times and extracting with 70-80% ethanol 2-3 times in sequence; specifically preferably: extracting with 90-100% ethanol 1-2 times, after solid-liquid separation, obtaining filter residue and filtrate; extracting the filter residue with 70-80% ethanol 2-3 times, obtaining filter residue and filtrate, and concentrating the filtrate; more preferably: extracting with 95% ethanol once, after solid-liquid separation, obtaining filter residue and filtrate; extracting the filter residue with 75% ethanol twice. In the present invention, the time for each extraction is preferably 20-30h, more preferably 24-26h; during each extraction process, stirring is performed every 6h; the temperature of the extraction is preferably room temperature; the method of solid-liquid separation is preferably suction filtration. In the present invention, the concentration is preferably reduced pressure concentration; the temperature of the concentration is preferably 30-60°C, more preferably 45-50°C; the present invention has no special requirements for the pressure of the reduced pressure concentration, and the common method used by those skilled in the art can be used.

[0051] After obtaining the alcohol extract, the present invention dissolves and resuspends the alcohol extract in distilled water to obtain a suspension; the suspension is mixed with butanol, extracted and concentrated to obtain a butanol phase extract. In the present invention, the butanol is preferably water-saturated n-butanol; the volume ratio of the butanol to the suspension is preferably 1:1-3; the number of extractions is preferably 3-5 times, more preferably 3-4 times.

[0052] In the present invention, the extraction temperature is preferably room temperature; the concentration is preferably consistent with the above-mentioned concentration, which will not be repeated here; the concentration is also preferably dried; and the drying is preferably freeze-drying.

[0053] After obtaining the butanol phase extract, the present invention performs adsorption resin column separation on the butanol phase extract, and obtains a 20% ethanol elution component and a 40% ethanol elution component after concentration. In the present invention, the adsorption resin in the adsorption resin column separation is preferably HP-20; the mass ratio of the adsorption resin and the butanol phase extract is preferably 8 to 16:1; the eluents used in the adsorption resin column separation are water, 20% ethanol and 40% ethanol in sequence; the water is preferably distilled water; during the elution process, the column volumes of the water, 20% ethanol and 40% ethanol are independently preferably 2 to 3BV. In the present invention, the 20% ethanol and 40% ethanol refer to 20% volume fraction ethanol and 40% volume fraction ethanol, and the remaining ethanol is volume fraction.

[0054] After obtaining the 20% ethanol elution component, the present invention performs a first chromatographic column separation on the 20% ethanol elution component, combines the same fractions, and concentrates to obtain components A1 to A10. In the present invention, the first chromatographic column separation is preferably ODS chromatographic column separation, and the filler is preferably ODS-A; the specification of the filler is preferably 12nm S-50μm; the chromatographic column separation preferably includes: adsorption by the filler, volatilization and removal of the solvent, and then dry column packing, and separation by medium pressure preparative chromatography. In the present invention, the 20% ethanol elution component is also preferably dissolved before the first chromatographic column separation; the dissolving reagent is preferably ethanol.

[0055] In the present invention, the eluents used for separation in the first chromatographic column are 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 50% ethanol and 100% ethanol, respectively; the column volume of each gradient is preferably 2 to 3 BV; in the separation in the first chromatographic column, when no chromatographic peak appears at 254 nm, the mobile phase with the next ethanol concentration is replaced for elution; the merging of the same elution fractions is preferably performed by merging the same fractions after detection by a photodiode array detector (PDAD) to obtain A1 to A10 fractions (10 elution fractions); the A3 fraction is concentrated to obtain the A3 component; the concentration is preferably consistent with the above-mentioned concentration.

[0056] After obtaining component A3, the present invention performs a first gel column chromatography separation on the component A3, combines the same fractions, and concentrates to obtain components A3a to A3s; performs a second chromatography column separation on the component A3c to obtain the glycoside compound of formula I; and performs a third chromatography column separation on the component A3e to obtain the glycoside compound of formula II.

[0057] In the present invention, the filler of the first gel column chromatography separation is preferably Sephadex LH-20; the mobile phase of the first gel column chromatography separation is water, preferably distilled water; the loading amount of the A3 component is preferably 1-5% of the gel column volume, more preferably 1-2%; during the first gel column chromatography separation process, each 1 / 20 column volume is preferably one fraction; after the first gel column chromatography separation, the same fractions are preferably combined after PDAD detection to obtain A3a~A3s fractions (19 elution fractions); the A3c fraction and the A3e fraction are concentrated to obtain A3c component and A3e component.

[0058] In the present invention, the second chromatographic column separation and the third chromatographic column separation are independently preferably ODS chromatographic column separation, and the filler is independently preferably ODS-AQ; the specification of the filler is preferably 250×20mm; 12nm; the elution mode in the second chromatographic column separation and the third chromatographic column separation is independently preferably isocratic elution; the eluent used in the second chromatographic column separation is preferably 20% methanol, and the detection wavelength is preferably 256nm; the eluent used in the third chromatographic column separation is 15% methanol, and the detection wavelength is preferably 256nm.

[0059] The 40% ethanol elution group is separated by a fourth chromatographic column, and the same fractions are combined and concentrated to obtain components B1 to B9. In the present invention, the method of separation by the fourth chromatographic column is preferably consistent with the method of separation by the first chromatographic column, which will not be repeated here; the eluents used for separation by the fourth chromatographic column are 20% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 60% ethanol, 80% ethanol and 100% ethanol in sequence;

[0060] After obtaining the B1 component, the present invention performs a second gel column chromatography separation on the B1 component, and obtains B1a to B1o components after concentration; the second gel column chromatography separation is preferably consistent with the first gel column chromatography separation, to obtain B1a to B1o fractions (15 elution fractions), and fraction B1a is concentrated to obtain component B1a.

[0061] After obtaining the B1a component, the present invention sequentially separates the B1a component by a fifth chromatographic column to obtain the glycoside compound of formula III and the glycoside compound of formula IV. In the present invention, the separation method of the fifth chromatographic column is preferably consistent with the separation method of the second and third chromatographic columns; the eluent used for the separation of the fifth chromatographic column is 20% methanol; and the detection wavelength is preferably 239 nm.

[0062] The present invention also provides the use of the glycoside compound described in the above technical solution in the preparation of a drug for treating diabetes, a drug for treating diabetic complications or a drug for neurodegenerative diseases.

[0063] In the present invention, the glycoside compounds of formula I are mainly involved in processes such as energy metabolism, inflammatory response, immune cell function, cell growth and survival, and cell response to hypoxia, and have therapeutic effects on diabetes and its complications; the glycoside compounds of formula II mainly act on mitochondrial respiratory chain complex I, regulate mitochondrial function and energy metabolism, and have therapeutic effects on neurodegenerative diseases; the glycoside compounds of formula III and formula IV focus on regulating cell metabolism and signal transduction, and also have therapeutic effects on nervous system diseases.

[0064] In order to further illustrate the present invention, the glycoside compounds provided by the present invention and their preparation methods and applications are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0065] Example 1

[0066] The bamboo stalks of Phyllostachys pubescens were collected, and the leaves were removed and the bamboo pieces were cut into bamboo pieces with a width of less than 3 cm, and the pieces were crushed into 20-40 meshes by a hammer mill; 53 kg of bamboo stalk powder was weighed, and 95% ethanol was used for extraction at room temperature at a solid-liquid ratio of 1:4 kg / L, and the mixture was filtered, and the obtained filter residue was extracted twice by 75% ethanol, and each extraction was performed for 24 hours. During the extraction process, the mixture was stirred every 6 hours, and the extract was concentrated under reduced pressure at 45° C. to obtain 2246 g of ethanol extract of Phyllostachys pubescens stalks;

[0067] The ethanol extract was dissolved and resuspended in an appropriate amount of distilled water, and extracted four times at a volume ratio of ethanol extract to water-saturated n-butanol of 1:1 kg / L. The n-butanol extract phase was concentrated under reduced pressure at 45°C, and freeze-dried to obtain 446 g of n-butanol phase extract;

[0068] The n-butanol phase extract was dissolved and resuspended with an appropriate amount of distilled water, separated with a macroporous resin HP-20, and eluted with distilled water (2BV), 20% ethanol (3BV) and 40% ethanol (3BV) as mobile phases in sequence, and the 20% ethanol eluate and the 40% ethanol eluate were concentrated under reduced pressure to obtain 61g of the 20% ethanol portion and 86g of the 40% ethanol portion;

[0069] The 20% ethanol portion was dissolved in ethanol, adsorbed by C18 filler ODS-A (12nm S-50μm), evaporated to remove the solvent, and then dry-packed on a column. The mixture was separated by medium-pressure preparative chromatography (Sepacore X10), and eluted with 5%, 10%, 15%, 20%, 25%, 30%, 50% and 100% ethanol (2BV) in sequence. When no chromatographic peak appeared at 254nm, the mobile phase was replaced with a mobile phase of the next concentration. After detection by a photodiode array detector (PDAD), the same fractions were combined to obtain A1 to A10, a total of 10 fractions. The A3 fraction was concentrated under reduced pressure to obtain 10.2g of A3.

[0070] A3 was dissolved in distilled water and separated by chromatography on a Sephadex LH-20 column. The loading amount was 1% of the volume of the gel column. Each 1 / 20 column volume was a fraction. After PDAD detection, the same fractions were combined to obtain a total of 19 fractions, which were represented by letters a to s. Fractions c and e were concentrated under reduced pressure to obtain 1022.0 mg A3c component and 588.8 mg A3e component, respectively.

[0071] A3c was dissolved in 10 mL of distilled water and prepared by semi-preparative liquid chromatography (Shimadzu), the chromatographic column was ODS-AQ (250×20 mm; 12 nm), the mobile phase was 20% methanol, isocratic elution, the detection wavelength was 256 nm, and repeated accumulation was performed to obtain 41.0 mg of the glycoside compound of formula I, and the purity of the glycoside compound of formula I was tested; the HPLC purity test conditions were: the chromatographic column was ODS-AQ (250×4.6 mm; 12 nm), 20% methanol isocratic, the flow rate was 1 mL / min, the full wavelength (200-400 nm), and the retention time was 8.5 min;

[0072] A3e was dissolved in 10 mL of distilled water and prepared by semi-preparative liquid chromatography (Shimadzu), the chromatographic column was ODS-AQ (250×20 mm; 12 nm), the mobile phase was 15% methanol, isocratic elution, the detection wavelength was 239 nm, and repeated accumulation was performed to obtain 5.8 mg of the glycoside compound of formula II, and the purity of the glycoside compound of formula II was tested; the HPLC purity test conditions were: the chromatographic column was ODS-AQ (250×4.6 mm; 12 nm), 15% methanol isocratic, the flow rate was 1 mL / min, the full wavelength (200-400 nm), and the retention time was 22.3 min;

[0073] The 40% ethanol portion was dissolved in ethanol, adsorbed by C18 filler ODS-A (12nm S-50μm), evaporated and removed the solvent, and then dry packed on a column, and separated by medium pressure preparative chromatography (Sepacore X10). Eluted with 20%, 30%, 35%, 40%, 45%, 50%, 60%, 80%, and 100% ethanol (3BV) in sequence. When no chromatographic peak appeared at 254nm, the mobile phase was replaced with the next concentration. After PDAD detection, the same fractions were combined to obtain B1 to B9, a total of 9 fractions, and the B1 fraction was concentrated under reduced pressure to obtain 42.0g of B1;

[0074] B1 was dissolved in distilled water and separated by chromatography on a Sephadex LH-20 column. The loading amount was 1% of the gel column volume. Each 1 / 20 column volume was a fraction. After PDAD detection, the same fractions were combined to obtain 15 fractions in total, which were represented by letters a to o. Fraction a was concentrated under reduced pressure to obtain 724.0 mg of B1a.

[0075] B1a was dissolved in 10 mL of distilled water and prepared by semi-preparative liquid chromatography (Shimadzu), the chromatographic column was ODS-AQ (250×20 mm; 12 nm), the mobile phase was 20% methanol, isocratic elution, the detection wavelength was 254 nm, and repeated cumulative preparation was performed to obtain 15.0 mg of the glycoside compound of formula III and 31.5 mg of the glycoside compound of formula IV, respectively. The purity of the glycoside compound of formula III and the glycoside compound of formula IV were tested; the HPLC purity test conditions were: the chromatographic column was ODS-AQ (250×4.6 mm; 12 nm), 20% methanol isocratic, the flow rate was 1 mL / min, the full wavelength (200-400 nm), and the retention times were 7.2 min and 6.4 min, respectively.

[0076] Test Example 1

[0077] The glycoside compounds of the present invention were characterized, and the results are as follows:

[0078] The chemical structural formula of the glycoside compound of formula I is:

[0079]

[0080] The NMR data of the glycoside compound of formula I are shown in Table 1:

[0081] Table 1 NMR data results of glycoside compounds of formula I (DMSO-d6)

[0082]

[0083]

[0084] From the results in Table 1, we can see that UV(H2O)λ max (log·ε)255.8nm HRESIMS m / z:567.1566[M+HCOO] - (calculated for 567.1567). High-resolution mass spectrometry and nuclear magnetic resonance data suggest that the molecular formula of the compound is C 21 H 30 O 15 , the unsaturation degree is 7. The H NMR spectrum data showed that the glycoside compound of formula I has two aromatic ring hydrogen signals δ H 7.49 (d, J = 1.9, 1H), δ H 7.28 (d, J = 1.9, 1H), the coupling constant is 1.9 Hz, indicating 4J coupling, which is a typical AB spin coupling system; the two anomeric hydrogen signals δ H 5.08 (d, J = 7.6, 1H), δ H 4.82 (d, J = 7.1, 1H), the coupling constants are 7.6 and 7.1 respectively, indicating that there are two D-glucose linked by β-glycosidic bonds; a methoxy signal 3.82 (s, 6H), indicating that it contains two methoxy groups. The C NMR spectrum data showed that compound 1 contains a carbonyl carbon signal δ C 165.7, six carbon signals on the benzene ring: δ C 152.9, δ C 150.4, δ C 139.7, δ C 125.1, δ C 111.6, δ C 108.2; two methoxyl signals δ C 52.3, δ C 56.5; δ C 102.9 and δ C 102.3 are respectively attributed to the anomeric carbon of the two β-D-glucose units. H 7.49) / C-7(δ C 165.7), H-2(δ H 7.28) / C-7(δ C 165.7), suggesting that the carbonyl carbon (C-7) is connected to the benzene ring (C-1); the two methoxyl hydrogen spectrum signals are respectively related to C-7 (δ C 165.7) and C-3 (δ C152.9), suggesting that the methoxy group is linked to C-7 and C-3, respectively; the two anomeric hydrogen signals (H-1" and H-1') are respectively linked to C-4 (δ C 139.7) and C-5 (δ C 150.4), it is speculated that the two β-D-glucose residues are connected to C-4 and C-5 on the benzene ring respectively. In summary, the glycoside compound of formula I is named methyl3-methoxybenzoate-4,5-di-O-β-D-glucopyranoside.

[0085] The chemical structural formula of the glycoside compound of formula II is:

[0086]

[0087] Table 2 NMR data results of glycoside compounds of formula II (DMSO-d6)

[0088]

[0089]

[0090] From the results in Table 2, we can see that UV(H2O)λ max (log·ε)274.7nm; HRESIMS m / z:457.1349[M+HCOO] - , (calculated for 457.1351). High-resolution mass spectrometry and nuclear magnetic resonance data suggest that the molecular formula of the compound is C 19 H 24 O 10 , the unsaturation degree is 8. The hydrogen spectrum data show that the glycoside compound of formula II contains a methoxy signal peak δ H 3.77 (s, 3H), three aromatic ring hydrogen signals: δ H 7.05 (d, 8.4 Hz, 1H), δ H 6.98 (d, 2.0 Hz, 1H), δ H 6.87 (dd, 8.4, 2.0 Hz, 1H), which belongs to a typical ABX spin coupling system. In addition, an anomeric hydrogen signal δ H 4.89 (d, 7.3 Hz, 1H), the coupling constant is 7.3, suggesting a β-glycosidic bond. Carbon spectrum data show that II glycoside compounds contain a carbonyl carbon signal δ C 178.8; 6 benzene ring carbon signals δ C 149.0, δ C 146.1, δ C 133.8, δ C 118.2, δC 115.1, δ C 110.5, a methoxyl signal δ C 55.7, δ C 100.0 is the anomeric carbon, and the remaining five carbon signals of the β-D-glucose unit are δ C 77.1, δ C 76.9, δ C 73.2, δ C 69.7 and δ C 60.7. At this point, the remaining 5 carbon signals need to be assigned to the remaining 2 unsaturations, which suggests that the II glycoside compounds may have a bridged ring structure. The key long-range correlation data of the HMBC spectrum show that H-1" (δ H 4.89) / C-4'(δ C 146.1), suggesting that the β-D-glucose unit is connected to C-4'; the methoxy group (δ H 3.77) and C-3'(δ C 149.0), suggesting that a methoxy group is connected to the C-3' position; H-3 (δ H 4.69) and C-2'(δ C 110.5) and C-6'(δ C 118.2), suggesting that C-3 is connected to C-1'. H 4.19, 3.95) and carbonyl carbon C-1 (δ C 178.8), suggesting that C-4 is connected to C-1 via a COC bond, while H-3 (δ H 4.69) and oxygen-containing carbon C-2 (δ C 70.3), suggesting that C-2 and C-3 are also connected by a COC bond. Combined with 1H-1H COSY data, H-2 (δ H 4.48, 4.35) / H-1a(δ H 3.12) / H-3a(δ H 3.59), indicating that C-2, C-1a, C-3a, and C-3 form a tetrahydrofuran ring; H-1a (δ H 3.12) / H-3a(δ H 3.59) / H-4(δ H4.19, 3.95), indicating that C-1a, C-3a, C-4 and C-1 form a γ-butyrolactone ring, and the two rings share C-1a and C-3a. Therefore, the planar structural formula of II glycoside compound is 3-(4'-O-β-D-glucopyranosyl-3'-methoxyphenyl)-5,6-dioxa-bicyclo-[3.3.0]octane-1-one. The results of circular dichroism showed that II glycoside compound had negative Cotton effect at 231.5nm and 274.5nm, and its absolute configuration was determined to be 1aS, 3aS, 3R, so it was named (1aS, 3aS, 3R)3-(4'-O-β-D-glucopyranosyl-3'-methoxyphenyl)-5,6-dioxa-bicyclo-[3.3.0]octane-1-one.

[0091] The chemical structural formula of the glycoside compound of formula III is:

[0092]

[0093] Table 3 NMR data results of glycoside compounds of formula III (DMSO-d6)

[0094]

[0095] From the results in Table 3, we can see that UV(H2O)λ max (log·ε)266.4nm; HRESIMS m / z:519.1714[M+HCOO] - , (calculated for 519.1719). High-resolution mass spectrometry and nuclear magnetic resonance data suggest that the molecular formula of the compound is C 21 H 30 O 12 The unsaturation degree is 7. The hydrogen spectrum data show that the glycoside compound of formula III contains three aromatic ring hydrogen signals δ H 7.47 (d, 2.1 Hz, 1H), δ H 7.60 (dd, 8.6, 2.1 Hz, 1H), δ H 7.23 (d, 8.6 Hz, 1H), which belongs to a typical ABX spin coupling system. H 3.82 (s, 3H) is the methoxy signal; δ H 1.07 (t, 7.2 Hz, 3H) and δ H 3.01 (q, 7.3 Hz, 2H) are located in the high field region, which are methyl and methine signals respectively. H 5.00 (d, 7.8 Hz, 1H) and δH 4.15 (d, 7.6 Hz, 1H) are two anomeric hydrogens, which can be preliminarily judged to be β-D-glucose and xylose according to the chemical shift and coupling constant. The carbon spectrum data showed that the glycoside compound of formula III contained 21 carbon signals, including a carbonyl δ C 199.1, one benzene ring δ C 150.4, δ C 148.6, δ C 130.6, δ C 122.4, δ C 114.6 and δ C 110.7, one methyl delta C 8.4, a methylene δ C 30.9, one methoxyl group δ C 55.1, two anomeric carbons δ C 103.8 and δ C 99.5; the remaining 9 carbon signals are located in the high field range of 60-80 ppm, which are oxygen-containing carbons and belong to the sugar unit. Together with the two anomeric carbons, there are 11 carbons in total. It can be seen that the glycoside compound of formula III contains hexose and pentose units, which is consistent with the results of hydrogen spectrum analysis. According to the HMBC spectrum, the connection position of each structural unit can be known, and the methoxy group (δ H 3.82) and C-3(δ C 148.6), suggesting that the methoxy group is connected to C-3; H-1'(δ H 5.00) and C-4(δ C 150.4), suggesting that the glucose unit is connected to C-4; H-1" (δ H 4.15) and C-6'(δ C 68.0), suggesting that xylose is linked to the C-6' position of glucose. H 1.07), H-8(δ H 3.01) are all related to C-7(δ C 199.1), combined with the 1H-1H COSY spectrum suggesting that H-9 is related to H-8, it can be seen that C-9 is connected to C-8 to form an ethyl group, and then connected to the carbonyl group (C-7) through a CC bond. In summary, the glycoside compound of formula III is 3-methoxy-propiophenone 4-O-(6'-β-D-xylopyranosyl)-β-D-glucopyranoside.

[0096] The chemical structural formula of the glycoside compound of formula IV is:

[0097]

[0098] Table 4 NMR data results of glycoside compounds of formula IV (DMSO-d6)

[0099]

[0100] From the results in Table 4, we can see that UV(H2O)λ max (logε)254.6nm; HRESIMS m / z:521.1511[M+HCOO] - , (calculated for 521.1512). High-resolution mass spectrometry and nuclear magnetic resonance data suggest that the molecular formula of the compound is C 20 H 28 O 13 The unsaturation degree is 7. The nuclear magnetic resonance hydrogen spectrum shows that the glycoside compound of formula IV contains three aromatic ring hydrogen signals δ H 7.47 (d, 2.0 Hz, 1H), δ H 7.56 (dd, 8.5, 2.0 Hz, 1H), δ H 7.24 (d, 8.5 Hz, 1H); two methoxy groups δ H 3.82 (s, 6H); two anomeric hydrogen δ H 4.13 (d, 7.6 Hz, 1H), δ H 4.97 (d, 7.7 Hz, 1H); the carbon NMR spectrum showed that the glycoside compound of formula IV contained 20 carbon signals, which can be attributed to a carbonyl δ C 166.0, two methoxyl groups C 52.0 and δ C 55.7, one benzene ring δ C 150.6, δ C 148.5, δ C 112.3, δ C 123.0, δ C 123.1 and δ C 114.8, two anomeric carbon δ C 99.7 and Delta C 104.0; the remaining 9 carbon signals are located in the high field 60-80ppm region, plus two anomeric carbons, a total of 11 carbon signals, respectively belonging to a hexose and a pentose. From the above analysis, it can be seen that the nuclear magnetic hydrogen spectrum and carbon spectrum data of the glycoside compound of formula IV are very similar to those of the glycoside compound of formula III. It is speculated that the structure of the glycoside compound of formula IV is similar to that of the glycoside compound of formula III. Combined with the HMBC spectrum analysis, the glycoside compound of formula IV has one less ethyl unit and one more methoxy unit than the glycoside compound of formula III. The methoxy group (δ H3.82) is related to C-7 (166.0), suggesting that the methoxy group is connected to the carbonyl group (C-7), and the rest of the structural information analysis results are the same as those of the glycoside compound of formula III. Therefore, the glycoside compound of formula IV is methyl3-methoxybenzoate 4-O-(6'-β-D-xylopyranosyl)-β-D-glucopyranoside.

[0101] Test Example 2

[0102] The results of virtual screening of the biological activity of the glycoside compounds of the present invention are as follows:

[0103] The reverse virtual screening technology based on the pharmacophore model and structure was comprehensively utilized to predict the biological activities of the glycoside compounds described in Formula I to Formula IV in Example 1 of the present invention. The 3D structural model of the glycoside compounds described in Formula I to Formula IV was constructed using Chemdraw 20.0, and the energy was minimized by the MM2 force field. The target prediction and screening were performed based on Pharm Mapper (z score>0.8), SEA (z score>10), Super-Pred (probability>60%; Model accuracy>80%), NetInfer (score>0.8), and SwissTargetPrediction databases to generate the target information of the glycoside compounds described in Formula I to Formula IV. The results of the above five databases were combined, and the duplicate targets were removed to obtain the predicted biological activity targets of the glycoside compounds described in Formula I to Formula IV.

[0104] The protein-protein interaction (PPIs) network was predicted using the STRING database, and the species was set to Homo sapiens to obtain protein interaction information. The topological index was calculated using Cytoscape (V3.10.2), and the protein interaction network was constructed based on the degree. The top 20 key targets were screened using the MCC algorithm, and the key subnetworks were obtained by calculating the indexes including betweenness centrality (BC), closeness centrality (CC), degree centrality (DC), eigenvector centrality (EC), and local average connectivity method (LAC) based on the MCODE algorithm. Among the key targets, AKT1 is the most critical target of the glycoside compound of Formula I, with an MCC value of 3.92E+08. In the key subnetwork, IL2 is the core target and the network center, and AKT1 is also in a core position in the subnetwork. By merging the key target set with the targets in the key sub-network and removing duplications, the core targets were obtained: NFKB1, SMAD3, GAPDH, STAT1, EPAS1, FGF2, IL2, AKT1, KIT, PTGS2, and HRAS.

[0105] The calculated core targets of the glycoside compounds of formula II, formula III and formula IV are: NDUFA10, NDUFB11, NDUFB9, NDUFAF1, NDUFC2, MT-ND1, NDUFB4, NDUFS3, NDUFAF2, NDUFB7, NDUFS4, NDUFS7, NDUFB10, NDUFS6, NDUFA2; GSK3B, PIK3CA, JAK2, FYN, ERBB2, FGF2, CDC42, EGFR, HRAS, KIT, RHOA, NFKB1, PIK3R1, STAT1; GAPDH, FGF2, KIT, PIK3R1, IL2.

[0106] Table 5 Topological values ​​of key target sites for protein interaction of glycoside compounds of formula I and glycoside compounds of formula II

[0107]

[0108]

[0109] Table 6 Topological values ​​of key target sites for protein interaction of glycoside compounds of formula III and glycoside compounds of formula IV

[0110]

[0111]

[0112] Table 7 Topological values ​​of key subnetworks of protein interactions of glycoside compounds of formula I

[0113] Target Points Node Status BC CC DC EC LAC IL2 11.74 Seed 1.05 0.72 11.00 2.00 9.09 MMP13 9.62 Clustered 0.00 0.64 8.00 2.00 7.00 CDC42 9.63 Clustered 1.98 0.72 11.00 2.00 8.55 RAF1 10.00 Clustered 0.75 0.64 8.00 2.00 6.25 LGALS3 9.70 Clustered 1.64 0.69 10.00 2.00 7.80 LCK 9.55 Clustered 2.30 0.72 11.00 2.00 8.36 RHOA 10.06 Clustered 1.51 0.69 10.00 2.00 7.80 NFKB1 9.66 Clustered 15.40 1.00 18.00 1.00 11.33 NFE2L2 10.52 Clustered 0.50 0.67 9.00 2.00 7.56 ATM 9.62 Clustered 0.40 0.64 8.00 2.00 6.50 SMAD3 10.51 Clustered 10.47 0.90 16.00 2.00 10.38 GAPDH 9.66 Clustered 15.40 1.00 18.00 1.00 11.33 STAT1 10.46 Clustered 7.11 0.86 15.00 2.00 10.53 EPAS1 9.99 Clustered 1.48 0.72 11.00 2.00 8.91 FGF2 10.39 Clustered 4.87 0.82 14.00 2.00 10.29 AKT1 9.66 Clustered 15.40 1.00 18.00 1.00 11.33 KIT 11.43 Clustered 4.57 0.78 13.00 2.00 9.23 PTGS2 10.04 Clustered 5.33 0.82 14.00 2.00 10.14 HRAS 10.07 Clustered 11.83 0.95 17.00 2.00 11.18

[0114] Table 8 Topological values ​​of key subnetworks of protein interactions of glycoside compounds of formula II

[0115]

[0116]

[0117] Table 9 Topological values ​​of key subnetworks of protein interactions of glycoside compounds of formula III

[0118]

[0119]

[0120] Table 10 Topological values ​​of key subnetworks of protein interactions of glycoside compounds of formula IV

[0121] Target Points Node Status BC CC DC EC LAC LGALS3 8.32 Seed 9.06 0.59 8.00 2.00 4.75 IL2 6.74 Clustered 133.61 0.77 19.00 2.00 5.58 AKT2 8.19 Clustered 14.57 0.59 9.00 3.00 3.78 ZAP70 7.64 Clustered 5.39 0.56 7.00 3.00 3.71 ACE 8.08 Clustered 0.62 0.57 7.00 2.00 5.43 LCK 7.42 Clustered 5.51 0.60 9.00 2.00 5.56 DPP4 7.42 Clustered 11.50 0.59 8.00 2.00 4.25 PIK3R1 7.78 Clustered 25.45 0.64 12.00 2.00 5.17 CCR1 7.15 Clustered 0.00 0.50 3.00 3.00 2.00 FGF2 6.89 Clustered 30.95 0.66 13.00 2.00 5.38 ARG2 7.00 Clustered 0.50 0.54 4.00 2.00 2.50 AOC3 7.00 Clustered 0.83 0.56 6.00 2.00 4.33 EPHA2 7.15 Clustered 2.07 0.53 5.00 3.00 2.80 CHUK 7.00 Clustered 0.56 0.53 3.00 2.00 1.33 SLC2A1 6.81 Clustered 1.17 0.55 5.00 2.00 3.20 NFE2L2 8.00 Clustered 1.17 0.51 4.00 3.00 2.00 NOS2 7.42 Clustered 8.74 0.57 7.00 2.00 3.14 CD209 7.00 Clustered 0.00 0.47 3.00 3.00 2.00 SELL 7.29 Clustered 17.04 0.60 9.00 2.00 4.22 KIT 8.05 Clustered 31.82 0.64 12.00 2.00 5.00 GAPDH 6.75 Clustered 210.17 0.90 24.00 2.00 6.33 CDK2 8.19 Clustered 5.37 0.56 6.00 2.00 2.67 MMP1 7.63 Clustered 3.40 0.59 8.00 2.00 5.50 AR 8.32 Clustered 4.04 0.55 6.00 3.00 3.33 ELANE 7.27 Clustered 1.83 0.54 5.00 3.00 2.80 RAC2 7.27 Clustered 7.39 0.57 8.00 3.00 4.25 SERPINE1 7.91 Clustered 14.10 0.61 10.00 2.00 4.80 MAPK12 7.27 Clustered 5.14 0.54 6.00 3.00 3.00

[0122] DAVID (Updated on April 17, 2024) was used to perform gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis on the predicted targets, and enrichment analysis based on the DisGeNET database was performed. GO entries and KEGG pathways with p-values ​​< 0.01 were considered significantly enriched, and the most critical biological activity information of glycoside compounds of Formula I to Formula IV was obtained.

[0123] Figure 1 The predicted target PPIs diagram, key subnetwork diagram and key target diagram of the glycoside compound of formula I are shown in FIG. Figure 1 a~c are respectively the predicted target PPIs graph, key subnetwork graph and key target graph, Figure 1 The results show that the glycoside compounds of formula I are mainly involved in biological processes such as single organism metabolism, small molecule metabolism, phosphorus metabolism, and phosphate-containing compound metabolism. They have biological functions such as catalytic activity, ion binding, small molecule binding, and carbohydrate derivative binding. The sites of action include extracellular exosomes, extracellular vesicles, extracellular organelles, vesicles, etc.

[0124] Figure 2 The GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula I are shown in FIG. Figure 2 a and b are the GO map and KEGG enrichment result map of the predicted target, respectively. Figure 2It can be seen that the action pathways of the glycoside compounds of formula I are metabolic pathways, cancer pathways, VEGF signaling pathways, starch and sucrose metabolism, etc. The results of the DisGeNET database enrichment analysis show that the diseases that the glycoside compounds of formula I may act on include myocardial ischemia, diabetes-experimental, vascular diseases, cerebral infarction, etc.

[0125] Figure 3 The predicted target PPIs diagram, key subnetwork diagram and key target diagram of the glycoside compound of formula II are shown in FIG. Figure 3 a~c are respectively the predicted target PPIs graph, key subnetwork graph and key target graph, Figure 3 The results show that the glycoside compounds of formula II are mainly involved in biological processes such as mitochondrial electron transfer, NADH to ubiquinone, mitochondrial ATP synthesis coupled proton transfer, energy coupled proton transport-electrochemical gradient decrease, etc., and have biological functions such as NADH dehydrogenase (ubiquinone) activity, NADH dehydrogenase (quinone) activity, NADH dehydrogenase activity, and oxidoreductase activity. The sites of action include respiratory chain complex I, mitochondrial respiratory chain complex I, NADH dehydrogenase complex, respiratory chain complex, etc.

[0126] Figure 4 The GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula II are shown in FIG. Figure 4 a and b are the GO map and KEGG enrichment result map of the predicted target, respectively. Figure 4 It can be seen that the action pathways are chemical carcinogenesis-reactive oxygen, retrograde endocannabinoid signaling, oxidative phosphorylation, thermogenesis, etc. The results of DisGeNET database enrichment analysis show that the diseases that the formula II glycoside compounds may act on include nicotinamide adenine dinucleotide coenzyme Q reductase deficiency, increased cerebrospinal fluid lactate, mitochondrial complex deficiency, mitochondrial myopathy, etc.

[0127] Figure 5 The predicted target PPIs map, key subnetwork map and key target map of the glycoside compound of formula III are shown in FIG. Figure 5 a~c are respectively the predicted target PPIs graph, key subnetwork graph and key target graph, Figure 5 The results show that the glycoside compounds of formula III are mainly involved in biological processes such as single-organism metabolism, reaction to organic matter, reaction to chemical substances, reaction to oxygen-containing compounds, etc., and have biological functions such as catalytic activity, small molecule binding, ion binding, and carbohydrate derivative binding. The sites of action include extracellular exosomes, vesicles, extracellular vesicles, extracellular organelles, etc.

[0128] Figure 6 The GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula III are shown in FIG. Figure 6a and b are the GO map and KEGG enrichment result map of the predicted target, respectively. Figure 6 It can be seen that the action pathways are metabolic pathways, neuroactive ligand-receptor interactions, cancer pathways, purine metabolism, etc. The results of the DisGeNET database enrichment analysis show that the diseases that the formula III glycoside compounds may act on include neuralgia, memory disorders, migraine diseases, central nervous system diseases, etc.

[0129] Figure 7 The predicted target PPIs diagram, key subnetwork diagram and key target diagram of the glycoside compound of formula IV are shown in FIG. Figure 7 a~c are the predicted target PPIs diagram, key subnetwork diagram and key target diagram, which are Figure 7 The results show that the glycoside compounds of formula IV are mainly involved in biological processes such as reactions to organic matter, reactions to chemical substances, single-organism metabolic processes, reactions to oxygen-containing compounds, etc., and have biological functions such as catalytic activity, small molecule binding, ion binding, and carbohydrate derivative binding. The sites of action include extracellular exosomes, extracellular vesicles, extracellular organelles, vesicles, etc. ( Figure 7 ).

[0130] Figure 8 The GO map and KEGG enrichment result map of the predicted targets of the glycoside compounds of formula IV are shown in FIG. Figure 8 a and b are the GO map and KEGG enrichment result map of the predicted target, respectively. Figure 8 It can be seen that the action pathways are neuroactive ligand-receptor interaction, metabolic pathway, cancer pathway, Rap1 signaling pathway, etc. ( Figure 8 ). The results of enrichment analysis of the DisGeNET database showed that the diseases that the glycoside compounds of Formula IV may act on include neuralgia, memory impairment, depression, unipolar depression, cognitive impairment, mood disorders, etc.

[0131] Based on the above analysis results, it can be seen that the glycoside compounds of formula I mainly play a potential therapeutic role in cardiovascular diseases, diabetes and its complications, tumors and other diseases by regulating energy metabolism, inflammatory response, immune cell function, cell growth and survival, and cell response to hypoxia. GO and KEGG enrichment analysis showed that the glycoside compounds of formula I mainly participated in metabolism-related biological processes, especially the metabolism of phosphates, carbohydrates and nucleotides, suggesting that they may affect cell function by regulating cell energy metabolism. The structural characteristics of the glycoside compounds of formula I also support this speculation, and their multiple hydroxyl and carbonyl groups may be involved in carbohydrate metabolism and phosphorylation reactions. The core target analysis further revealed the possible mechanism of action of the glycoside compounds of formula I. For example, core targets such as nuclear factor NF-κBp105 subunit, SMAD family member 3, signal transducer and activator of transcription 1 and interleukin 2 are involved in inflammatory response and immune regulation, suggesting that the glycoside compounds of formula I may affect the inflammatory response and immune cell function by regulating these targets, thereby playing a therapeutic role in inflammatory diseases and immune-related diseases. Core targets such as glyceraldehyde-3-phosphate dehydrogenase, protein kinase B and HRas oncogene are involved in cell proliferation and apoptosis, suggesting that glycoside compounds of formula I may affect cell growth and survival by regulating these targets, thereby playing a therapeutic role in diseases such as tumors. Core targets such as endothelial PAS domain protein 1 and hypoxia-inducible factor 1-α are involved in the response of cells to hypoxia, suggesting that glycoside compounds of formula I may affect the adaptability of cells under hypoxic conditions by regulating these targets, thereby playing a therapeutic role in ischemic diseases. The results of pathway analysis are mutually confirmed by core target analysis. The vascular endothelial growth factor signaling pathway, advanced glycation end product receptor signaling pathway and hypoxia-inducible factor-1 signaling pathway involved in glycoside compounds of formula I are all related to angiogenesis, inflammatory response and cell response to hypoxia, and the key molecules in these pathways are the above core targets. DisGeNET database analysis predicts that glycoside compounds of formula I may act on myocardial ischemia, diabetes and its complications, and tumors, which is consistent with the results of pathway analysis and target analysis. For example, the vascular endothelial growth factor signaling pathway is related to angiogenesis, myocardial ischemia and vascular diseases; the advanced glycation end product receptor signaling pathway is related to diabetic complications; the hypoxia-inducible factor-1 signaling pathway is related to tumor growth and metastasis; and the core targets nuclear factor NF-κB p105 subunit, SMAD family member 3, signal transducer and activator of transcription 1 and interleukin 2 are also related to inflammatory response and immune regulation, which are consistent with the predicted disease types.

[0132] The reverse virtual screening analysis results of formula II glycosides revealed their potential key roles in cellular energy metabolism and mitochondrial function, suggesting their potential application value in the treatment of various mitochondrial-related diseases. The results of GO and KEGG enrichment analysis showed that formula II glycosides were mainly involved in the mitochondrial electron transport chain, ATP synthesis and purine nucleotide biosynthesis, which constitute the core of cellular energy metabolism. Gene ontology enrichment analysis further predicted that formula II glycosides may have nicotinamide adenine dinucleotide dehydrogenase activity, which is a key component of mitochondrial respiratory chain complex I, responsible for starting the electron transport chain and ultimately driving the generation of ATP. Its action sites are mainly concentrated in the mitochondrial respiratory chain complex, especially complex I, which is the starting step of the mitochondrial electron transport chain and is also a susceptible link for mitochondrial dysfunction. Core target analysis further confirmed the effect of formula II glycosides on mitochondrial function. The core targets such as NADH dehydrogenase subunit A10, NADH dehydrogenase subunit B11, NADH dehydrogenase subunit B9, NADH dehydrogenase assembly factor 1, NADH dehydrogenase subunit C2, and mitochondrial encoded NADH dehydrogenase 1 are all subunits or assembly factors of mitochondrial respiratory chain complex I, suggesting that formula II glycoside compounds may directly act on complex I to affect its assembly, activity or stability, thereby regulating the function of mitochondrial electron transport chain. The results of pathway analysis are mutually confirmed with the core target analysis. The "chemical carcinogenesis-reactive oxygen" and "oxidative phosphorylation" pathways involved in formula II glycoside compounds are closely related to oxidative stress and mitochondrial dysfunction, suggesting that formula II glycoside compounds may counteract oxidative stress by improving mitochondrial function and reducing reactive oxygen generation, thereby producing therapeutic effects on related diseases. In addition, formula II glycoside compounds are also associated with neurodegenerative disease pathways such as Parkinson's disease, Alzheimer's disease and Huntington's disease, and the occurrence and development of these diseases are often accompanied by mitochondrial dysfunction and increased oxidative stress. This suggests that the glycoside compounds of formula II may play a therapeutic role in neurodegenerative diseases by protecting mitochondrial function and delaying neuronal damage. DisGeNET database analysis predicts that the glycoside compounds of formula II may act on a variety of mitochondrial diseases, such as nicotinamide adenine dinucleotide coenzyme Q reductase deficiency, mitochondrial complex deficiency, mitochondrial myopathy, etc., all of which are related to mitochondrial dysfunction. The glycoside compounds of formula II may directly act on the mitochondrial respiratory chain complex I to regulate mitochondrial function and energy metabolism, thereby playing a therapeutic role in a variety of mitochondrial-related diseases, especially neurodegenerative diseases.

[0133] The reverse virtual screening analysis results of the glycoside compounds of formula III revealed their complex biological activities, suggesting that they may play a therapeutic role in various diseases through multi-target and multi-pathway regulatory mechanisms. The results of gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the glycoside compounds of formula III are mainly involved in metabolism-related biological processes, and have functions such as catalytic activity, small molecule binding, ion binding, as well as signal transduction-related functions such as signal transducer activity and receptor activity. This suggests that the glycoside compounds of formula III may affect cell function by regulating cell metabolism and signal transduction. The core target analysis further supports this speculation. The core targets of the formula III glycoside compounds include glycogen synthase kinase-3β, phosphatidylinositol 3-kinase catalytic subunit α, Janus kinase 2, Fyn proto-oncogene tyrosine kinase, Erb-B2 receptor tyrosine kinase 2, fibroblast growth factor 2, cell division cycle protein 42, epidermal growth factor receptor, HRas oncogene, KIT proto-oncogene receptor tyrosine kinase, Rho-related GTP binding protein RhoA, nuclear factor NF-κB p105 subunit, phosphatidylinositol 3-kinase regulatory subunit 1 and signal transducer and activator of transcription 1, etc. These targets are involved in a variety of cell signaling pathways, such as Ras signaling pathway, Rap1 signaling pathway, cAMP signaling pathway, EGFR tyrosine kinase inhibitor resistance pathway, etc., and affect cell growth, proliferation, differentiation, apoptosis, immune regulation and inflammatory response. The biological activity of the formula III glycoside compounds is closely related to their potential therapeutic application value. DisGeNET database analysis predicts that the glycoside compounds of formula III may act on a variety of diseases, such as neuralgia, memory impairment, diabetes, cerebral infarction, pneumonia, myocardial ischemia, endothelial dysfunction, breast cancer, hypertension, central nervous system diseases and vascular diseases. These diseases are associated with the core targets of the glycoside compounds of formula III and the signal pathways involved. For example, the glycoside compounds of formula III may affect tumor growth and angiogenesis by regulating targets such as fibroblast growth factor 2 and KIT proto-oncogene receptor tyrosine kinase, thereby exerting a therapeutic effect on tumor diseases such as breast cancer. The glycoside compounds of formula III may also affect neurotransmitter receptors and cell signal transduction by regulating neuroactive ligand-receptor interactions, Rap1 signaling pathways and cAMP signaling pathways, thereby exerting a therapeutic effect on neuralgia, memory impairment and central nervous system diseases.

[0134] The reverse virtual screening analysis results of the glycosides of formula IV revealed their wide range of biological activities, suggesting that they may play a therapeutic role in a variety of diseases through multi-target and multi-pathway regulatory mechanisms. The results of gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the glycosides of formula IV are mainly involved in metabolism-related biological processes, and have functions such as catalytic activity, small molecule binding, ion binding, as well as signal transducer activity, purinergic receptor activity and other functions related to signal transduction. This suggests that the glycosides of formula IV may affect cell function by regulating cell metabolism and signal transduction. The core target analysis further supports this speculation. The core targets of the glycosides of formula IV are the same as those of the glycosides of formula III, including glyceraldehyde-3-phosphate dehydrogenase, fibroblast growth factor 2, KIT proto-oncogene receptor tyrosine kinase, phosphatidylinositol 3-kinase regulatory subunit 1 and interleukin 2. These targets are involved in a variety of cell signaling pathways, such as the Ras signaling pathway, the Rap1 signaling pathway, the calcium signaling pathway, the HIF-1 signaling pathway, the EGFR tyrosine kinase inhibitor resistance pathway, etc., and affect processes such as cell growth, proliferation, differentiation, apoptosis, immunoregulation and inflammatory response. The biological activity of the glycoside compounds of formula IV is closely related to their potential therapeutic application value. DisGeNET database analysis predicts that the glycoside compounds of formula IV may act on a variety of diseases, such as diabetes, vascular diseases, neuralgia, myocardial ischemia, lung diseases, cerebral infarction, pneumonia, endothelial dysfunction, memory impairment, hypercholesterolemia, head and neck cancer, reperfusion injury, idiopathic pulmonary hypertension, malignant head and neck tumors, lymphoma, depression, cognitive impairment, mood disorders, etc. These diseases are associated with the core targets of the glycoside compounds of formula IV and the involved signaling pathways. For example, the glycoside compounds of formula IV may affect tumor growth and angiogenesis by regulating targets such as fibroblast growth factor 2 and KIT proto-oncogene receptor tyrosine kinase, thereby exerting a therapeutic effect on tumor diseases such as head and neck cancer and lymphoma. The glycoside compounds of formula IV may also affect neurotransmitter receptors, cell signal transduction and inflammatory response by regulating neuroactive ligand-receptor interactions, calcium signaling pathways and AGE-RAGE signaling pathways, thereby playing a therapeutic role in neuralgia, memory disorders, diabetes and its complications. It is worth noting that the glycoside compounds of formula III and the glycoside compounds of formula IV have the same core targets, but there are certain differences in the pathways involved and the predicted disease types. This suggests that although the two may act on the same target, their mechanisms of action and downstream effects may be different, resulting in differences in the therapeutic effects on different diseases.

[0135] In summary, based on the current target prediction and enrichment analysis, it can be inferred that the glycoside compounds of formula I have a wide range of biological activities and may have therapeutic potential for a variety of diseases, especially cardiovascular diseases, diabetes and its complications, cancer, etc., which provides an important reference for the subsequent pharmacodynamic studies of the compound. The reverse virtual screening analysis results of the glycoside compounds of formula II strongly suggest that they have the potential to regulate mitochondrial function and energy metabolism, and may have potential therapeutic value for a variety of mitochondrial-related diseases, especially neurodegenerative diseases, cardiomyopathy and metabolic diseases. The reverse virtual screening analysis results of the glycoside compounds of formula III suggest that they have the potential to regulate cell metabolism and signal transduction, and may affect cell function by regulating cell metabolism and signal transduction, thereby exerting a therapeutic effect on a variety of diseases, especially nervous system diseases, cardiovascular diseases, tumors and metabolic diseases, and have potential therapeutic value. Reverse virtual screening analysis revealed that the glycoside compounds of formula IV may exert their biological activity by intervening in cell metabolism and signal transduction, thereby exerting a therapeutic effect on a variety of diseases. Its potential therapeutic application directions include metabolic diseases, cardiovascular diseases and tumors.

[0136] GO and KEGG pathway enrichment analysis of glycosides of formula I and III showed that both were involved in metabolism-related biological processes. Glycosides of formula I focused more on the metabolism of phosphates, carbohydrates, and nucleotides, while glycosides of formula III focused more on the metabolism of organic matter and organic nitrogen compounds. Structural analysis showed that glycosides of formula I had multiple hydroxyl groups and carbonyl groups, suggesting that they may be involved in carbohydrate metabolism and phosphorylation reactions. Glycosides of formula III contained aromatic rings and carbonyl groups, suggesting that they may be involved in lipid metabolism and redox reactions. Pathway analysis showed that glycosides of formula I were involved in VEGF, AGE-RAGE, and HIF-1 signaling pathways, suggesting that they may play a role by regulating angiogenesis, inflammatory responses, and cellular responses to hypoxia. Glycosides of formula III were involved in neuroactive ligand-receptor interactions, Rap1 signaling pathways, and cAMP signaling pathways, suggesting that they may play a role by regulating neurotransmitter receptors and immune cell functions. DisGeNET analysis predicted that the glycoside compounds of formula I may act on cardiovascular diseases, diabetes and its complications, and tumors, while the glycoside compounds of formula III may act on nervous system diseases, immune system diseases, and tumors, etc.

[0137] The glycoside compounds of formula IV and formula III are similar in structure and core targets. Both may affect cell function by regulating cell metabolism and signal transduction, thereby playing a therapeutic role in a variety of diseases. The glycoside compounds of formula IV and formula III may participate in lipid metabolism, redox reaction, and cell signal transduction. The core targets of both include glyceraldehyde-3-phosphate dehydrogenase, fibroblast growth factor 2, KIT proto-oncogene receptor tyrosine kinase, phosphatidylinositol 3-kinase regulatory subunit 1 and interleukin 2, etc., and participate in Ras, Rap1, hypoxia-inducible factor-1 and calcium signaling pathways. The glycoside compounds of formula IV are more inclined to regulate the calcium signaling pathway and the advanced glycation end product receptor signaling pathway, and play a therapeutic role in metabolic diseases such as diabetes and its complications, while the glycoside compounds of formula III are more inclined to regulate the cAMP signaling pathway and the prolactin signaling pathway, and play a therapeutic role in nervous system diseases and immune system diseases.

[0138] In summary, the glycoside compounds of formula I to formula IV all exhibit a wide range of biological activities and may play a therapeutic role in a variety of diseases through multi-target and multi-pathway regulatory mechanisms. The glycoside compounds of formula I are mainly involved in energy metabolism, inflammatory response, immune cell function, cell growth and survival, and cell response to hypoxia, suggesting that they have potential therapeutic value for cardiovascular diseases, diabetes and its complications, tumors, etc. The glycoside compounds of formula II mainly act on mitochondrial respiratory chain complex I, regulate mitochondrial function and energy metabolism, and may have therapeutic potential for mitochondrial-related diseases such as neurodegenerative diseases. The glycoside compounds of formula III and formula IV focus more on regulating cell metabolism and signal transduction, and may play a therapeutic role in nervous system diseases, cardiovascular diseases, tumors and metabolic diseases.

[0139] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A glycoside compound, characterized in that: It has the following structure:

2. The method for preparing the glycoside compound according to claim 1, characterized in that: The following steps are involved: The plant of Phyllostachys genus is mixed with ethanol, extracted and concentrated to obtain an alcohol extract; Dissolving and resuspending the alcohol extract in distilled water to obtain a suspension; mixing the suspension with butanol, extracting and concentrating the mixture to obtain a butanol phase extract; The butanol phase extract is subjected to adsorption resin column separation, and after concentration, a 20% ethanol elution component and a 40% ethanol elution component are obtained; the eluents used in the adsorption resin column separation are water, 20% ethanol and 40% ethanol, respectively; The 20% ethanol elution component is separated by a first chromatographic column, and the same fractions are combined and concentrated to obtain components A1 to A10; the eluents used for the first chromatographic column separation are 5% ethanol, 10% ethanol, 15% ethanol, 20% ethanol, 25% ethanol, 30% ethanol, 50% ethanol and 100% ethanol, respectively; The A3 component is subjected to a first gel column chromatography separation, and the same fractions are combined and concentrated to obtain components A3a to A3s; the mobile phase of the first gel column chromatography separation is water; The A3c component is separated by a second chromatographic column to obtain the glycoside compound of formula I, and the eluent used for the second chromatographic column separation is 20% methanol; the A3e component is separated by a third chromatographic column to obtain the glycoside compound of formula II, and the eluent used for the third chromatographic column separation is 15% methanol; The 40% ethanol elution group is subjected to a fourth chromatographic column separation, and the same fractions are combined and concentrated to obtain components B1 to B9; the eluents used for the fourth chromatographic column separation are 20% ethanol, 30% ethanol, 35% ethanol, 40% ethanol, 45% ethanol, 50% ethanol, 60% ethanol, 80% ethanol and 100% ethanol, respectively; The B1 component is subjected to a second gel column chromatography separation, and after concentration, components B1a to B1o are obtained; the mobile phase of the second gel column chromatography separation is water; The B1a component is sequentially separated by a fifth chromatographic column to obtain the glycoside compound of formula III and the glycoside compound of formula IV. The eluent used in the fifth chromatographic column separation is 20% methanol.

3. The preparation method according to claim 2, characterized in that: The extraction comprises: extracting with 90-100% ethanol for 1-2 times and extracting with 70-80% ethanol for 2-3 times in sequence, each extraction lasting 20-30 hours; the solid-liquid ratio of the Phyllostachys plant to ethanol is 1:3-5kg / L.

4. The preparation method according to claim 2, characterized in that: The extraction times are 3 to 5 times; the butanol is water-saturated n-butanol; and the volume ratio of the butanol to the suspension is 1:1 to 3.

5. The preparation method according to claim 2, characterized in that: The adsorption resin in the adsorption resin column separation is HP-20.

6. The preparation method according to claim 2, characterized in that: The first chromatographic column separation and the fourth chromatographic column separation are independently ODS chromatographic column separations, and the filler is ODS-A.

7. The preparation method according to claim 2, characterized in that: The fillers of the first gel column chromatography separation and the second gel column chromatography separation are independently Sephadex LH-20.

8. The preparation method according to claim 2, characterized in that: The second chromatographic column separation, the third chromatographic column separation and the fifth chromatographic column separation are independently ODS chromatographic column separations, and the filler is ODS-AQ.

9. Use of the glycoside compound according to claim 1 in the preparation of a drug for treating diabetes, a drug for treating diabetic complications or a drug for neurodegenerative diseases.

Images