Preparation method and application of a new glycoside component in valeriana jatamansi

Abstract

The application discloses a preparation method and application of a new glycoside component in Metaplexis japonica, and the compound is separated from a Metaplexis japonica extract. The Metaplexis japonica root extract is separated by using extraction, macroporous adsorption resin impurity removal, HW-40F gel column chromatography and high performance liquid chromatography, and a new glycoside compound is obtained. The inhibitory activity of the separated compound on diabetes related target alpha-glucosidase and protein tyrosine phosphatase 1B (PTP1B) is tested, and the experimental results show that the compound has good inhibitory activity on the two enzymes, and molecular docking experiments also show that the compound has good binding energy with the two target proteins alpha-glucosidase and protein tyrosine phosphatase 1B. Therefore, the compound can be used for preparing medicines for preventing, delaying or treating diseases mediated by alpha-glucosidase or PTP1B, in particular, type II diabetes and obesity, or as a lead compound of the medicines.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biology and medicine, and relates to the separation and purification and application of glycosides from the roots of Onosma paniculatum Maxim., in particular to a method for separating and purifying new glycoside compounds from the roots of Onosma paniculatum Maxim., and a preparation method thereof, and the inhibitory activities of the new glycoside compounds on α-glucosidase and protein tyrosine phosphatase 1B (PTP1B). BACKGROUND

[0002] Onosma paniculatum Maxim. Eupatorium chinense Onosma paniculatum Maxim. (O.P.), a plant of the Asteraceae family, has a root like a bundle of hairs, and is also known as Du Xugong, a commonly used Chinese herbal medicine in the Tujia and Miao ethnic groups in western Hubei and Hunan provinces and Guangdong province. The chemical components of Onosma paniculatum Maxim. include flavonoids, terpenes, benzofuran, alkaloids, essential oils, etc. Onosma paniculatum Maxim. is recorded in Fujian Pharmacopoeia as having the effects of soothing the liver and relieving depression, opening the chest and relieving the diaphragm, regulating the menstrual cycle and promoting blood circulation, and relieving swelling and pain. In Jiangsu and Zhejiang provinces, Onosma paniculatum Maxim. is used by the public to make tea by cutting the roots into pieces, which can effectively control the increase of blood sugar. With the development of modern pharmacological research, it has been shown that Onosma paniculatum Maxim. has various pharmacological activities such as anti-inflammatory, antiviral, antitumor, and antibacterial activities, but there is less research on its antidepressant and hypoglycemic activities.

[0003] α-glucosidase inhibitors can promote the entry of part of undigested carbohydrates into the small intestine, stimulate the secretion of a large amount of glucagon-like peptide 1 (GLP-1) in the site, and this change in the level of GLP-1 secretion helps to further control blood sugar. In addition, α-glucosidase inhibitors can also effectively improve and prevent the occurrence and development of some T2DM complications, and have no obvious hepatorenal toxic side effects. Therefore, α-glucosidase inhibitors have good efficacy for Asian T2DM patients who mainly eat carbohydrates, and show unique advantages in the control of T2DM and its complications (Wang Meiyun. Design, synthesis, activity and mechanism of α-glucosidase and PTP1B dual-target inhibitors [D]. Tianjin Medical University, 2017.).

[0004] Protein tyrosine phosphatase 1B (PTP1B), the product of PTPN1 (protein tyrosine phosphatase, non-receptor type 1) gene, is a non-transmembrane protein tyrosine phosphatase. PTP1B plays a key role in the negative regulation of insulin and leptin signaling pathways, and maintains the balance of tyrosine protein phosphorylation with protein tyrosine kinase (Protein Tyrosine Kinase, PTK). PTP1B knockout mice show enhanced insulin sensitivity, improved blood glucose levels and obesity resistance (E S N, P T C, S M, et al. Potential role of glycogen synthase kinase-3 in skeletal muscle insulin resistance of type 2 diabetes. [J]. Diabetes, 2000, 49(2): 263-71.), and their islet morphology measurements show that the proliferation level of islet β cells is increased, and the isolated islet secretes high levels of insulin (O T J, Jacques E, R M J. Protein tyrosine phosphatase 1B inhibitors for diabetes. [J]. Nature reviews. Drug discovery, 2002, 1(9): 696-709.), which indicates that PTP1B is an important physiological regulator of insulin release in islets. PTP1B is induced by mediating ER stress to cause insulin resistance, which is an important way to develop insulin resistance. PTP1B is a direct target of miR122, and abnormal regulation of liver miR122 can induce PTP1B through the c-Jun N-terminal kinase 1 (JNK1)-hepatocytenuclear factor 4α axis, and then promote liver insulin resistance (Yazhen X, Xin X. Glucagon receptor mediates calcium signaling by coupling to G alpha q / 11 and G alpha i / o in HEK293 cells. [J]. Journal of receptor and signal transduction research, 2009, 29(6): 318-25.).PTP1B can increase the level of high-fat metabolism by strengthening leptin signal, thus reducing body weight, therefore, PTP1B inhibitor can reduce or even reverse the resistance of obese patients to insulin and leptin, which is of great significance for the treatment of T2DM patients, especially obese T2DM patients. Therefore, the simultaneous inhibition of the two target points of α-glucosidase and PTP1B can reduce postprandial blood glucose on one hand, and enhance insulin sensitivity on the other hand, reduce the load of pancreatic beta cells, and jointly maintain the stability of blood glucose level in the body, and can reduce the body weight of obese diabetic patients by increasing the level of leptin in the body. SUMMARY

[0005] The first object of the present application is to provide a kind of radix eupatorium glycosides compound and its preparation method.The second object is to provide its use in the preparation of drugs for preventing or treating α-glucosidase or protein tyrosine phosphatase PTP1B mediated diseases.

[0006] The compound with inhibitory activity of α-glucosidase or PTP1B has the following structural formula:

[0007]

[0008] Eupbenzofuranside C.

[0009] The technical scheme of the present application is the use of a kind of glycosides compound in radix eupatorium in the preparation of drugs for preventing, delaying or treating α-glucosidase or protein tyrosine phosphatase PTP1B mediated diseases.

[0010] The preparation method of the radix eupatorium glycosides compound comprises the following steps:

[0011] A1, solvent extraction: after the radix eupatorium is crushed, it is sequentially extracted by ethanol, petroleum ether, ethyl acetate and n-butanol, and then the n-butanol extract is concentrated to obtain extract a;

[0012] B1, macroporous adsorption resin impurity removal of extract a: extract a is dissolved by ultrasonic in deionized water, macroporous adsorption resin (D101 type) is soaked in deionized water, washed, wet loaded, and then adsorbed overnight, eluted with ethanol / deionized water system, and the eluate is concentrated under reduced pressure to obtain extract b;

[0013] C1, the extract b is dissolved with methanol, and the extract is roughly separated by reverse phase silica gel column chromatography, wet loaded, dry loaded, and gradient eluted with methanol / deionized water system, and the eluate is collected;

[0014] D1, the eluate is separated by HW-40F gel column chromatography to obtain the separation product;

[0015] E1, the separation product is subjected to reverse silica gel column chromatography for preliminary separation, and the obtained fragment is subjected to semi-preparative HPLC to prepare compound 1.

[0016] In step C1, gradient elution is performed using 10% methanol / 90% water, 15% methanol / 85% water, 20% methanol / 80% water, 30% methanol / 70% water, 40% methanol / 60% water, 60% methanol / 40% water, 80% methanol / 20% water and 100% methanol, and the eluate in the part of 30% methanol / 70% water is collected, or the eluate in the part of 40% methanol / 60% water is collected, or the eluate in the part of 60% methanol / 40% water is collected.

[0017] The eluate in step D1 is subjected to gel column chromatography HW-40F and separated by methanol-water in a volume fraction of 10-25:70-85 to obtain the separation product.

[0018] In step E1, the separation product is subjected to reverse silica gel column chromatography and separated by methanol-water in a volume fraction of 55-70:55-70.

[0019] In the semi-preparative HPLC, the mobile phase is acetonitrile-water 60:40, and the flow rate is 1-2 mL / min.

[0020] A medicine for treating diabetes and obesity, the medicine comprising the prepared compound 1.

[0021] A compound for inhibiting the activity of α-glucosidase or PTP1B, the compound comprising the prepared compound 1.

[0022] A medicine for preventing, delaying or treating diabetes and obesity by inhibiting the activity of α-glucosidase or PTP1B, the medicine comprising the prepared compound 1.

[0023] The compound 1 is combined with α amino acids in the active site of the glucosidase protein to form a site containing hydrophobic interaction and hydrogen bond interaction;

[0024] The compound 1 is combined with amino acids in the active site of the PTP1B protein to form a site containing hydrophobic interaction and hydrogen bond interaction.

[0025] The compound or medicine of the present application also comprises a pharmaceutically acceptable excipient.

[0026] Compared with the prior art, the present application has at least one of the following beneficial effects:

[0027] 1. The compound 1 of the present application has significant α-glucosidase or PTP1B inhibitory activity, and there is no any public report about the compound having α-glucosidase or PTP1B inhibitory activity and application in related diseases.

[0028] 2. The compound having α-glucosidase or PTP1B inhibitory activity isolated from the n-butanol extract of the root of the plant Daphne giraldii is found for the first time, and has broad application prospect for development into a drug for preventing or treating α-glucosidase or PTP1B mediated related diseases, such as diabetes, obesity. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 is a chemical structure diagram and a two-dimensional correlation diagram of the compound 1.

[0030] Figure 2 is a 1H-NMR spectrum of the compound 1. 1

[0031] Figure 3 is a 13C-NMR spectrum of the compound 1. 13

[0032] Figure 4 is a DEPT135 spectrum of the compound 1.

[0033] Figure 5 is a HSQC spectrum of the compound 1.

[0034] Figure 6 is a 1H-1H COSY spectrum of the compound 1. 1 1

[0035] Figure 7 is a HMBC spectrum of the compound 1.

[0036] Figure 8 is a NOESY spectrum of the compound 1.

[0037] Figure 9 is an experimental CD spectrum of the compound 1.

[0038] Figure 10 is a TD-DFT theoretical calculation ECD spectrum of the compound 1.

[0039] Figure 11 is an analysis diagram of the interaction of the compound 1 with α-glucosidase protein (wherein, A: 3D binding mode of the compound 1 with α-glucosidase protein; B: amino acid residue binding mode of the compound 1 with α-glucosidase protein; C: 2D binding mode of the compound 1 with α-glucosidase protein).

[0040] Figure 12 ​​​​Figure for analysis of the interaction of compound 1 with PTP1B protein, (wherein, A: 3D binding mode of compound 1 with PTP1B protein; B: amino acid residue binding mode of compound 1 with PTP1B protein; C: 2D binding mode of compound 1 with PTP1B). DETAILED DESCRIPTION

[0041] The following examples are provided to further explain the present application, and it is evident that the examples described are only a part of all the embodiments of the present application, but not all. Based on the examples in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present application.

[0042] Example 1

[0043] Isolation, preparation and structure analysis of one glycoside compound from the roots of Eupatorium fortunei Hara

[0044] The isolation and preparation method comprises the following steps:

[0045] Step A, solvent extraction: dry and crush the roots of Eupatorium fortunei Hara, extract 3 times with 95% ethanol under room temperature, concentrate by rotary evaporator, disperse the extract in water, and extract with petroleum ether, ethyl acetate and n-butanol in turn, each for 3 times. Concentrate the n-butanol extract to obtain extract a;

[0046] Step B, macroporous adsorption resin purification of extract a: take extract a, add 300 g of deionized water to dissolve under ultrasonic, soak 1000 g of macroporous adsorption resin (D101 type) with deionized water for 6 h, then column, wash with deionized water for at least three column volumes, put under solvent to be higher than the filler surface about 2 cm, wet sample, stand overnight adsorption, elute with 50% ethanol / 50% deionized water, wash until the eluent is colorless, and concentrate the eluent under reduced pressure to obtain extract b.

[0047] Step C, column chromatography: Take the above extract b, dissolved with a small amount of methanol, added to 90 g of reversed-phase silica gel (200-300 mesh) for mixing; take 1200 g of reversed-phase silica gel, soaked with 10% methanol / 90% deionized water, and use wet method to pack the column, with a column volume of 1 L, use 10% methanol / 90% deionized water to flush at least three column volumes, until the solvent level in the column is about 2 cm higher than the filler level, then use dry method to load, and after loading, plug in appropriate amount of defatted cotton as a protective layer, combine with high performance liquid analysis results, use methanol-water system to elute, and preliminarily separate the extract b. Use 10% methanol / 90% water, 15% methanol / 85% water, 20% methanol / 80% water, 30% methanol / 70% water, 40% methanol / 60% water, 60% methanol / 40% water, 80% methanol / 20% water and 100% methanol for gradient elution, each gradient elution at 2 column volumes, to obtain 8 fractions (Fr. A-H).

[0048] Use thin layer chromatography, high performance liquid chromatography and HPLC-DAD scanning analysis, it is shown that the 40% methanol / 60% water part Fr. E is the component where the target compound is located.

[0049] Step D, take the 40% methanol / 60% water part Fr. E, use gel column chromatography HW-40F (methanol / water=2:8) to perform isocratic elution, to obtain component Fr. E (1-17).

[0050] Step E, take Fr. E8, after preliminary separation by reversed-phase silica gel column chromatography (methanol / water=5:5), to obtain compound 33 (10.8 mg) and 4 fragments (Fr. E8-1-4), and separate compound 1 (28.9 mg) from Fr. E-8-2 by semi-preparative HPLC separation (acetonitrile-water 60:40, 2 mL / min, YMC C18 column).

[0051] Structure analysis of compound 1:

[0052] Nuclear magnetic resonance spectrum data and mass spectrum data are as follows: compound 1: yellow solid. Optical rotation data [α] 25 D -36.7° (c 0.10, methanol), UV (methanol) λmax=247, 278 nm; from high resolution mass spectrometry HR-ESI-MS: m / z 405.1532 [M+Na] + (C 19 H 26 NaO8, the calculated value is: 405.1525), the molecular formula is C 19 H 26 O8, the unsaturation degree is 8. Analyze the structure of compound 1 by 1The H-NMR spectrum revealed that the compound contained three methyl groups (proton signal). δ H 1.51 (s, H-13), 1.51 (s, H-14) and 1.41 (d, J =6.5Hz, H-11) and 1 olefin proton signal δ H 6.62 (d, J =0.8 Hz, H-3). From compound 1 13 The C-NMR and DEPT135 spectra show 19 carbons, including 3 methyl carbon signals. δ C 29.4 (C-13), 29.4 (C-14), and 25.2 (C-11); two methylene carbon signals δ C 61.7 (Glc-6'); 8 olefin carbon signals δ C 164.8 (C-2), 154.1 (C-8), 137.3 (C-5), 128.2 (C-9), 122.9 (C-6), 119.2 (C-4), 111.0 (C-7), 100.4 (C-3); 2 oxygen-carbon signals δ C 68.1 (C-12) and 74.1 (C-10). Compound 1 1 H-NMR δ H 3.89 (d, J = 7.5 Hz, H-1') indicates the presence of one terminal carbon, and the spectral data contains a set of glucose... 1 H-NMR and 13 C-NMR spectral signal ( δ C From 99.9, 77.4, 77.2, 74.1, 70.8, and 61.7, it can be seen that compound 1 contains one glycoside. Hydrolysis of the compound with 2.0 M HCl and HPLC analysis confirmed that it contains one D-glucose molecule. 1 H-NMR shows the coupling constant of the terminal proton signal at δH 3.89 ( J = 7.5 Hz)

[0053] NOESY analysis showed no correlation between H-1' and H-6', indicating that glucose has a β-configuration. The aglycone moiety of this compound was compared with that of literature compounds. 1 H-NMR and 13 Comparison of C-NMR spectra (Bohlmann F, Ziesche J, King RM,et al . Neue melampolide aus Smallanthus fruticosus[J]. Phytochemistry,1980, 19(5): 973-974.), C-10 of the literature compound ( δ C 68.7) becomes ( ) in compound 1. δ C (74.1) It was speculated that the glycosyl group might be attached at this position, which was confirmed by the relationship between H-1' and C-10 in the HMBC spectrum. Comprehensive analysis of the two-dimensional spectrum of compound 1 showed that its aglycone structure was completely identical to that of the literature compound. Thus, the planar structure of compound 1 was obtained.

[0054] The absolute configuration at position 10 was compared with the experimental ECD spectrum (which showed a negative cotton effect at 210 nm and a positive cotton effect at 280 nm) calculated using TD-DFT theory. Figure 10 It was determined to be in the S configuration, and after a search, it was found to be a new compound, named Eupbenzofuranside C.

[0055] Table 1. Comparison of Compound 1 with that of the literature compound 1 H-NMR and 13 Comparison of C-NMR data (solvent DMSO- d 6)

[0056] Example 2: Inhibitory activity test of compound 1 against α-glucosidase and PTP1B

[0057] Experimental methods

[0058] α-Glucosidase Activity Assay

[0059] The method for α-glucosidase assay was modified based on the method of Tao et al. (Biomedical chromatography: BMC.2013, 27(2): 148-155). The p-nitrophenyl-β-galactopyranoside method was used, in which pNPG, as a substrate, was decomposed into pNP and glucose under the catalysis of α-glucosidase. pNP showed a yellow color and strong UV absorption at 405 nm. Acarbose was used as a positive control. The amount of product after sample addition was measured to determine the inhibition rate of the sample. 80 μL of sample (butanol extract of *Euphorbia lathyris* or compound 1) solution (PBS buffer, pH 7.2) was added to each well of a 96-well plate. α- Glucosidase (2 U / mL, PBS buffer pH 7.2), mixed well. After incubation at 37 ℃ for 15 min, 5 mmol / L pNPG 20 μL was added to start the reaction. After incubation for 15 min, 80 μL 1 mol / L Na2CO3 was added to stop the reaction. In the blank control group, 1% PBS 80 μL was added instead of the sample solution. The absorbance (A) at 405 nm was used to quantify the amount of pNPG released. The enzyme activity group (enzyme 20 μL + reaction buffer 80 μL + substrate 20 μL), enzyme blank group (reaction buffer 100 μL + substrate 20 μL), sample group (sample 80 μL + enzyme 20 μL + substrate 20 μL), sample blank group (sample 80 μL + reaction buffer 20 μL + substrate 20 μL) were set up, with acarbose solution as the positive control, and the inhibition rate was calculated according to the formula: inhibition rate = 1 - (A sample - A sample blank) / (A enzyme activity - A enzyme blank).

[0060] PTP1B enzyme activity determination

[0061] The determination method of PTP1B was carried out according to the literature (Funct. Foods, 2018, 41:232-239; Chinese Journal of Pharmacy, 2019, 54(3): 510-513.). The sample 10 μL (n-butanol extract of Huazelan or compound 1) was added to 170 μL of reaction buffer solution (consisting of 50 mM citric acid (pH 7.4), 50 mM NaCl, 2 mM dithiothreitol (DTT), 1.1 mM EDTA), 20 μL of recombinant PTP1B solution (1 mg / mL, 1 μL) was mixed in each well, the reaction mixture was preheated using a 37°C block heater for 15 min, 10 μL of reaction substrate pNPP (33 mM) was added, and the reaction was carried out at 37°C for 15 min, NaOH solution (10 μL, 0.1 M) was added to stop the reaction. The absorbance was recorded at 405 nm. The enzyme activity group (enzyme 20 μL + reaction buffer solution 170 μL + substrate 10 μL), enzyme blank group (reaction buffer 190 μL + reaction substrate 10 μL), sample group (sample 170 μ + enzyme 20 μL + reaction substrate 10 μL), sample blank group (sample 170 μL + reaction buffer solution 20 μL + reaction substrate 10 μL) were set up according to the above steps, with sodium orthovanadate solution and oleanolic acid solution as the positive control.

[0062] The inhibition rate was calculated according to the following formula: inhibition rate = 1 - (A sample - A sample blank) / (A enzyme activity - A enzyme blank)

[0063] Experimental results: The n-butanol extract of E. wallichii and compound 1 were tested for α-glucosidase and PTP1B enzyme inhibition activity, and the results are shown in Tables 2-3:

[0064] Table 2. Results of α-glucosidase inhibition activity of compound 1

[0065]

[0066] The test results show that the above-mentioned ginsenoside compounds from the roots of E. wallichii have good α-glucosidase inhibition activity.

[0067] Table 3. Results of PTP1B enzyme inhibition activity of compound 1

[0068]

[0069] The test results show that the above-mentioned ginsenoside compounds from the roots of E. wallichii have good α-glucosidase inhibition activity.

[0070] Example 3: Docking of compound 1 with two protein target molecules

[0071] Compound 1 in this docking was constructed using ChemDraw, and then imported into Chem3D software to optimize and minimize energy using the MM2 module, and saved as an sdf format file as a ligand molecule for molecular docking. Then, it was imported into Pymol and Autodock software for optimization processing, and exported as a PDBQT file. The α-glucosidase (PDB ID: 5ZCE) and PTP1B (PDB ID: 5QG3) protein structures were obtained from the RCSB database (https: / / www.rcsb.org / structure / 5ZCE; https: / / www.rcsb.org / structure / 5QG3) https: / / www.rcsb.org / ), and the protein structures were processed by removing water molecules, removing ligands, adding hydrogen, etc. in Pymol and Autodock platforms, and exported as PDBQT files. The processing and optimization of molecular docking were completed by the Grid module in the Autodock software. The PDBQT file in Example 2 was imported into the software, and the box was wrapped around the protein, and then molecular docking was performed. In addition, the complex of the protein and the small molecule was visualized and analyzed using Pymol.

[0072] Docking results

[0073] Molecular docking results

[0074] Table 4. Docking results of compound 1 with target proteins

[0075] Target protein Compound Binding energy (kcal / mol) α - glucosidases 1 -8.5 PTP1B 1 -21.4

[0076] Compound and protein interaction analysis

[0077] In this experiment, compound 1 was docked withα Molecular docking was performed between the compound and the target protein PTP1B. The docking results showed that the compound and the target protein had a good binding affinity and a high degree of match. Figure 11 and Figure 12 The binding energies of all compounds were less than -6 kcal / mol. The complexes formed by the docking compounds and proteins were visualized using Pymol 2.1 software to obtain the binding patterns. Based on the binding patterns, the amino acid residues binding between the compounds and the protein pockets were clearly visible. Compound 1 and... α - The PHE-144 amino acid at the active site of the glucosidase protein forms a hydrophobic interaction and forms multiple hydrogen bond interactions with amino acids ASP-60, ILE-143, GLN-256, THR-409, and ARG-411.

[0078] Compound 1 forms multiple hydrophobic interactions with TYR-46 and VAL-49 amino acids at the active site of PTP1B protein, and multiple hydrogen bond interactions with TYR-46, ASP-48, ASP-181, and GLN-266 amino acids; it has strong binding ability and plays an important role in anchoring small molecules in the protein pocket.

[0079] The above detailed embodiments provide a specific description of the analytical methods involved in this invention. It should be noted that the above description is only intended to help those skilled in the art better understand the methods and ideas of this invention, and is not intended to limit the scope of the invention. Without departing from the principles of this invention, those skilled in the art can make appropriate adjustments or modifications to this invention, and such adjustments and modifications should also fall within the protection scope of this invention.

Claims

1. A ginseng root glycoside compound, characterized in that, The structural formula is as follows: 。 2. The method for separating and extracting the root glycosides of Eupatorium fortunei according to claim 1, characterized in that, The method comprises the following steps: A1, solvent extraction: the roots of Eupatorium Fortunei Hance are crushed and then sequentially subjected to ethanol extraction, petroleum ether extraction, ethyl acetate extraction, and n-butanol extraction, and the n-butanol extract is concentrated to obtain an extract a; B1, macroporous adsorption resin impurity removal of the extract a: the extract a is dissolved in deionized water by ultrasonic, macroporous adsorption resin is soaked in deionized water and then column-mounted, washed, and then column-mounted by wet method, and then adsorbed overnight, and then eluted by an ethanol / deionized water system, and then the eluate is concentrated under reduced pressure to obtain an extract b; C1, the extract b is dissolved in methanol, and then subjected to column chromatography by using reverse phase silica gel, column-mounted by wet method, column-mounted by dry method, and then gradient eluted by using a methanol / deionized water system, and then the eluate is collected, and then gradient eluted by using 10% methanol / 90% water, 15% methanol / 85% water, 20% methanol / 80% water, 30% methanol / 70% water, 40% methanol / 60% water, 60% methanol / 40% water, 80% methanol / 20% water, and 100% methanol, and then the eluate in the part of 30% methanol / 70% water, or the eluate in the part of 40% methanol / 60% water, or the eluate in the part of 60% methanol / 40% water is collected; D1, the eluate is separated by HW-40F gel column chromatography to obtain a separation product, and the eluate is separated by gel column chromatography HW-40F by using methanol-water with a volume fraction of 10-25:70-85 to obtain the separation product; E1, the separation product is preliminarily separated by reverse phase silica gel column chromatography, and then the fragment is prepared by semi-preparative HPLC to obtain the Eupatorium Fortunei Hance root glycoside compound; The reverse phase silica gel column chromatography is separated by using methanol-water with a volume fraction of 55-70:55-70; In the semi-preparative HPLC, the mobile phase is acetonitrile-water 60:40, and the flow rate is 1-2 mL / min.

3. A medicament for treating a disease including diabetes or obesity, characterized by, The medicine comprises the compound of claim 1, and the medicine is prepared by inhibiting the activity of alpha-glucosidase or PTP1B to realize the preparation of a medicine for treating diabetes or obesity.

4. The medicament according to claim 3, characterized in that, The compound of claim 1 by binding to α - amino acids of the active site of the glucosidase protein, forming a site containing hydrophobic interactions and hydrogen bond interactions; The compound of claim 1 is combined with amino acids in the active site of PTP1B protein to form a site containing hydrophobic interaction and hydrogen bond interaction.

5. The medicament according to any one of claims 3-4, characterized in that, The medicine further comprises a pharmaceutically acceptable excipient.

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