Preparation method and application of novel glycoside component in eupatorium chinense

Abstract

The invention discloses a preparation method and application of a novel glycoside component in eupatorium chinense. The compound is separated from an eupatorium chinense extract. The eupatorium chinense root extract is separated by using an extraction method, macroporous adsorption resin impurity removal, HW-40F gel column chromatography, high performance liquid chromatography and other methods to obtain a new glycoside compound. The inhibitory activity of the separated compound on diabetes related targets alpha-glucosidase and protein tyrosine phosphatase 1B (PTP1B) is tested, the experimental result shows that the compound has good inhibitory activity on two enzymes, and the molecular docking experiment also shows that the compound has good inhibitory activity on the two enzymes. The compound has good binding energy with two target protein alpha-glucosidase and protein tyrosine phosphatase 1B. Therefore, the compound can be used for preparing medicines for preventing, delaying or treating alpha-glucosidase or PTP1B-mediated diseases, especially type II diabetes and obesity, or can be used as a lead compound of the medicines.

Description

Technical Field

[0001] The present invention belongs to the field of biology and medicine technology, and relates to the separation, purification and application of glycoside compounds from the roots of Herba Cynoglossi, and specifically to a separation, purification and preparation method of a new glycoside compound from the roots of Herba Cynoglossi, and its inhibitory activity on α-glucosidase and protein tyrosine phosphatase 1B (PTP1B). Background Art

[0002] Hua Zelan Eupatorium chinense . L), is a plant of the genus Eupatorium in the Asteraceae family. Its roots are like whiskers, so it is also known as Duoxugong. It is a commonly used Chinese herbal medicine in western Hubei, the Tujia ethnic group in western Hunan, and Guangdong Province. Its chemical components include flavonoids, terpenes, benzofurans, alkaloids, essential oils, etc. The "Fujian Pharmacopoeia" records that Huazelan has the effects of "soothing the liver and relieving depression, opening the chest and relieving diaphragm, regulating menstruation and blood circulation, and reducing swelling and relieving pain". In the Jiangsu and Zhejiang areas, the folks use Huazelan roots cut into pieces to make tea, which can effectively control the increase of blood sugar. With the development of modern pharmacological research, it has been shown that it has multiple pharmacological activities such as anti-inflammatory, antiviral, anti-tumor, and antibacterial, but its antidepressant and hypoglycemic pharmacological activities are less studied.

[0003] α-glucosidase inhibitors can promote the entry of some undigested carbohydrates into the small intestine, stimulating the secretion of glucagon-like peptide 1 (GLP-1) in this area. This change in the secretion level of GLP-1 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 without obvious liver and kidney toxicity. Therefore, α-glucosidase inhibitors have a good therapeutic effect on Asian T2DM patients who mainly eat carbohydrates, and show unique advantages in controlling T2DM and its complications. (Wang Meiyan. Design, synthesis, activity and mechanism of action of dual-target inhibitors of α-glucosidase and PTP1B [D]. Tianjin Medical University, 2017.).

[0004] Protein tyrosine phosphatase 1B (PTP1B) is a product encoded by the PTPN1 (protein tyrosine phosphatase, non-receptor type 1) gene and is a non-transmembrane protein tyrosine phosphohydrolase. PTP1B plays a key negative regulatory role in the insulin and leptin signaling pathways, and works with protein tyrosine kinase (PTK) to maintain the balance of tyrosine protein phosphorylation in the body. PTP1B knockout mice show enhanced insulin sensitivity, improved blood glucose levels and obesity resistance (ESN, PTC, SM, 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 pancreatic islet morphology measurement shows increased proliferation of pancreatic β cells, and the level of insulin secretion of isolated islets is high (OTJ, Jacques E, RMJ. Protein tyrosinephosphatase 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 pancreatic islets. PTP1B causes insulin resistance by mediating ER stress, which is an important pathway for the development of insulin resistance. PTP1B is a direct target of miR122. Abnormal regulation of hepatic miR122 can induce PTP1B through the c-Jun N-terminal kinase 1 (JNK1)-hepatocyte nuclear factor 4α axis, thereby promoting hepatic 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 fat metabolism by strengthening leptin signaling, thereby reducing body weight. Therefore, PTP1B inhibitors may 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 T2DM patients with obesity. Therefore, simultaneous inhibition of α-glucosidase and PTP1B can reduce postprandial blood sugar on the one hand, and enhance insulin sensitivity and reduce the load on pancreatic β cells on the other hand, jointly maintaining the stability of blood sugar levels in the body, and can reduce the body weight of obese diabetic patients by increasing leptin levels in the body. Summary of the invention

[0005] The first purpose of the present invention is to provide a glycoside compound from the root of Herba Cyperi and a preparation method thereof. The second purpose is to provide a use of the glycoside compound in the preparation of a drug for preventing or treating diseases mediated by α-glucosidase or protein tyrosine phosphatase PTP1B.

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

[0007] Eupbenzofuranside C.

[0008] The technical scheme of the present invention is to use a glycoside compound in the root of Huazelan in the preparation of drugs for preventing, delaying or treating diseases mediated by α-glucosidase or protein tyrosine phosphatase PTP1B.

[0009] The preparation method of the Huazelan glycoside compound comprises the following steps: A1. Solvent extraction: The root of Huazelan is crushed and extracted with ethanol, petroleum ether, ethyl acetate and n-butanol in sequence, and the n-butanol extract is concentrated to obtain extract a; B1. Extract a macroporous adsorption resin impurity removal: extract a is added with deionized water and ultrasonically dissolved, macroporous adsorption resin (D101 type) is soaked in deionized water and loaded onto the column, rinsed, wet loaded, left to stand overnight for adsorption, eluted with ethanol / deionized water system, and the eluate is concentrated under reduced pressure to obtain extract b; C1. Dissolve the extract b in methanol, and use reversed-phase silica gel column chromatography to crudely separate the extract. Use wet column packing, dry sample loading, and use a methanol / deionized water system for gradient elution to collect the eluate. D1, taking the eluate and subjecting it to HW-40F gel column chromatography to obtain a separated product; E1. The separated product was subjected to preliminary separation by reverse silica gel column chromatography, and the obtained fragments were prepared by semi-preparative HPLC to obtain compound 1.

[0010] In step C1, 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 are used for gradient elution, and the eluate in the 30% methanol / 70% water portion, or the eluate in the 40% methanol / 60% water portion, or the eluate in the 60% methanol / 40% water portion is collected.

[0011] The eluate in step D1 was separated by gel column chromatography HW-40F with methanol-water at a volume fraction of 10-25: 70-85 to obtain a separated product.

[0012] In step E1, the separated product is separated by reverse silica gel column chromatography with methanol-water at a volume fraction of 55-70:55-70.

[0013] In the semi-preparative HPLC, the mobile phase is acetonitrile-water 60:40, and the flow rate is 1-2 mL / min.

[0014] A medicine for treating diabetes and obesity, comprising the prepared compound 1.

[0015] A compound for inhibiting the activity of alpha-glucosidase or PTP1B, comprising the prepared compound 1.

[0016] A drug for preventing, delaying or treating diabetes and obesity is prepared by inhibiting the activity of alpha-glucosidase or PTP1B, and the drug comprises the prepared compound 1.

[0017] The compound 1 is prepared by α - The amino acids in the active site of the glucosidase protein combine to form a site containing hydrophobic interactions and hydrogen bond interactions; The compound 1 combines with the amino acids in the active site of the PTP1B protein to form a site containing hydrophobic interactions and hydrogen bond interactions.

[0018] The compound or drug of the present invention further includes a pharmaceutically acceptable excipient.

[0019] Compared with the prior art, the present invention has at least one of the following beneficial effects: 1. Compound 1 of the present application has significant α-glucosidase or PTP1B inhibitory activity, and there is no public report on the α-glucosidase or PTP1B inhibitory activity of the compound and its application in related diseases.

[0020] 2. For the first time, a compound with the ability to inhibit α-glucosidase or PTP1B activity was isolated from the n-butanol extract of the root of Huazelan. This has broad application prospects for the development of drugs that can prevent or treat α-glucosidase or PTP1B-mediated diseases, such as diabetes and obesity. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Chemical structure diagram and two-dimensional correlation diagram of compound 1.

[0022] Figure 2 For compound 1 1 H-NMR.

[0023] Figure 3 For compound 1 13 C-NMR.

[0024] Figure 4 This is the DEPT135 spectrum of compound 1.

[0025] Figure 5 is the HSQC spectrum of compound 1.

[0026] Figure 6 For compound 1 1 H- 1 H COSY spectrum.

[0027] Figure 7 is the HMBC spectrum of compound 1.

[0028] Figure 8 is the NOESY spectrum of compound 1.

[0029] Fig. 9 This is the experimental CD spectrum of compound 1.

[0030] Fig.10 This is the ECD spectrum calculated by TD-DFT theory for compound 1.

[0031] Fig.11 This is an analysis diagram of the interaction between compound 1 and α-glucosidase protein (wherein, A: 3D binding mode of compound 1 and α-glucosidase protein; B: amino acid residue binding mode of compound 1 and α-glucosidase protein; C: 2D binding mode of compound 1 and α-glucosidase protein).

[0032] Fig.12 This is an analysis diagram of the interaction between compound 1 and PTP1B protein (wherein, A: 3D binding mode of compound 1 and PTP1B protein; B: amino acid residue binding mode of compound 1 and PTP1B protein; C: 2D binding mode of compound 1 and PTP1B). DETAILED DESCRIPTION

[0033] The following implementation cases are used to further explain the present invention. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present application.

[0034] Example 1 Isolation, preparation and structural elucidation of a glycoside compound from the root of Huazelan The separation and preparation method comprises the following steps: Step A, solvent extraction: the root of Huazelan was dried and crushed, and refluxed and extracted with 95% ethanol for 3 times at room temperature. After concentration by rotary evaporator, the extract was dispersed in water, and extracted with petroleum ether, ethyl acetate, and n-butanol in sequence, and each was repeated three times. The n-butanol extract was concentrated to obtain extract a; Step B, impurity removal of extract a with macroporous adsorption resin: take extract a, add 300 g deionized water and ultrasonically dissolve it, soak 1000 g macroporous adsorption resin (type D101) in deionized water for 6 hours and then load it onto the column, rinse with deionized water for at least three column volumes, lower the solvent to about 2 cm above the filler surface, wet load, let stand overnight for adsorption, elute with 50% ethanol / 50% deionized water, rinse until the eluent is colorless, and concentrate the eluent under reduced pressure to obtain extract b.

[0035] Step C, column chromatography crude fractionation: Take the above extract b, dissolve it with a small amount of methanol, add it to 90g reverse phase silica gel (200-300 mesh), and mix the sample; weigh 1200g reverse phase silica gel, soak it with 10% methanol / 90% deionized water, use wet column packing, the column volume is 1 L, rinse with 10% methanol / 90% deionized water for at least three column volumes until the solvent level in the column is about 2 cm higher than the filler surface, use dry method to load the sample, after loading the sample, plug in an appropriate amount of absorbent cotton as a protective layer, combine the results of high performance liquid phase analysis, use methanol-water system to elute, and perform preliminary separation of extract b. Gradient elution was performed with 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, respectively, with each gradient washing for 2 column volumes to obtain 8 fractions (Fr.AH).

[0036] Thin layer chromatography, high performance liquid chromatography and HPLC-DAD scanning analysis showed that the 40% methanol / 60% water portion Fr.E was the component where the target compound was located.

[0037] Step D: Take the 40% methanol / 60% water fraction Fr.E and perform isocratic elution on a gel column chromatography HW-40F (methanol / water = 2:8) to obtain fractions Fr.E (1-17).

[0038] Step E: Fr. E8 was initially separated by reverse silica gel column chromatography (methanol / water = 5:5) to obtain compound 33 (10.8 mg) and four fragments (Fr. E8-1~4). Compound 1 (28.9 mg) was obtained from Fr. E-8-2 by semi-preparative HPLC separation (acetonitrile-water 60:40, 2 mL / min, YMC C18 column).

[0039] Structural analysis of compound 1: The data of NMR spectrum and mass spectrum 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; by high resolution mass spectrometry HR-ESI-MS: m / z 405.1532 [M+Na] + (C 19 H 26 NaO 8 , calculated value: 405.1525), molecular formula is C 19 H 26 O 8 The unsaturation degree is 8. 1 H-NMR spectrum shows that the compound has 3 methyl groups (proton signal δ H 1.51 (s, H-13), 1.51 (s, H-14) and 1.41 (d, J = 6.5 Hz, H-11)) and 1 olefin proton signal δ H 6.62 (d, J = 0.8 Hz, H-3). From the 13 C-NMR spectrum and DEPT135 spectrum show that there are 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 C164.8 (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-linked carbon signals δ C 68.1 (C-12) and 74.1 (C-10). 1 H-NMR δ H 3.89 (d, J = 7.5 Hz, H-1') indicates the presence of one terminal carbon. There is a set of glucose in the spectral data. 1 H-NMR and 13 C-NMR spectral signal ( δ C 99.9, 77.4, 77.2, 74.1, 70.8, 61.7), it can be seen that compound 1 has 1 glycoside compound. The compound was hydrolyzed with 2.0 M HCl and HPLC analysis confirmed that it contained 1 D-glucose. 1 H-NMR shows that the coupling constant of the terminal proton signal at δH 3.89 ( J = 7.5 Hz) NOESY showed that H-1' and H-6' had no correlation, indicating that the configuration of glucose was β configuration. 1 H-NMR and 13 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) in compound 1 becomes ( δ C 74.1), it is speculated that the sugar group may be connected at this position, which is confirmed by the relationship between H-1' and C-10 in the HMBC spectrum. Comprehensive analysis of the two-dimensional spectrum of compound 1 shows that its aglycone structure is exactly the same as that of the reference compound. Thus, the planar structure of compound 1 was obtained.

[0040] The absolute configuration of position 10 is compared with the ECD spectrum calculated by TD-DFT theory and the experimental ECD spectrum (negative cotton effect at 210 nm and positive cotton effect at 280 nm) ( Fig.10), which was determined to be S configuration, and was found to be a new compound after retrieval and named Eupbenzofuranside C.

[0041] Table 1. Comparison of compound 1 with reference compounds 1 H-NMR and 13 C-NMR data comparison (solvent DMSO- d 6 )

[0042] Example 2: Test of the inhibitory activity of compound 1 on α-glucosidase and PTP1B Experimental methods α-Glucosidase activity assay The α-glucosidase assay was modified according to the method of Tao et al. (Biomedical chromatography: BMC. 2013, 27(2): 148-155). The p-nitrophenyl-β-pyranogalactoside method was used. pNPG was used as a substrate and was decomposed into pNP and glucose under the catalysis of α-glucosidase. pNP was yellow and had strong ultraviolet absorption at 405 nm. Acarbose was used as a positive control to detect the amount of product after adding the sample to determine the inhibition rate of the sample. 80 μL of sample (n-butanol extract of Huazelan or compound 1) solution (PBS buffer at pH 7.2) and 20 μL of α -glucosidase (2 U / mL, pH 7.2 PBS buffer) was added and mixed thoroughly. After incubation at 37 °C for 15 min, 20 μL of 5 mmol / L pNPG was added to start the reaction. After incubation for 15 min, 80 μL of 1 mol / L Na 2 CO 3 Terminate the reaction. Add 80 μL of 1% PBS to the blank control group instead of the sample solution. The absorbance (A) at 405 nm was used to quantify the release of pNPG. Set up 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), and use acarbose solution as the positive control. The inhibition rate is calculated as follows: inhibition rate = 1-(A sample-A sample blank) / (A enzyme activity-A enzyme blank).

[0043] PTP1B enzyme activity assay The determination method of PTP1B was carried out according to the literature (Funct. Foods, 2018, 41:232-239; Acta Pharmaceutica Sinica, 2019, 54(3): 510-513.). 10 μL of sample (H. huazelan n-butanol extract or compound 1) was added to 170 μL of reaction buffer solution (composed of 50 mM citric acid (pH 7.4), 50 mM NaCl, 2 mM dithiothreitol (DTT), 1.1 mM EDTA), and 20 μL of recombinant PTP1B solution (1 mg / mL, 1 μL) was added to each well. The reaction mixture was preheated for 15 min using a block heater at 37°C, and 10 μL of reaction substrate p-nitrophenyl phosphate pNPP (33 mM) was added. The reaction was reacted at 37°C for 15 min, and NaOH solution (10 μL, 0.1 M) was added to stop the reaction. The absorbance was recorded at 405 nm. According to the above steps, 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 μL + enzyme 20 μL + reaction substrate 10 μL), and sample blank group (sample 170 μL + reaction buffer solution 20 μL + reaction substrate 10 μL) were set up, and sodium orthovanadate aqueous solution and oleanolic acid solution were used as positive controls.

[0044] The inhibition rate was calculated according to the following formula: Inhibition rate = 1-(A sample-A sample blank) / (A enzyme activity-A enzyme blank) Experimental results: The α-glucosidase and PTP1B enzyme inhibition activities of the n-butanol extract of Huazelan and compound 1 were tested. The results are shown in Tables 2~3: Table 2. Results of the inhibition of α-glucosidase activity by compound 1

[0045] The test results show that the above-mentioned glycoside compounds from the roots of Huazelan have good α-glucosidase inhibitory activity.

[0046] Table 3. Results of compound 1 inhibiting PTP1B enzyme activity

[0047] The test results show that the above-mentioned phenylglycoside compounds from the root of Huazelan have certain PTP1B enzyme inhibitory activity.

[0048] Example 3: Docking of Compound 1 with two protein target molecules The compound 1 for this docking was constructed using ChemDraw, and then imported into Chem3D software for optimization and energy minimization using the MM2 module, and saved as an sdf format file as the ligand molecule for molecular docking. It was then imported into Pymol and Autodock software for optimization and exported as a PDBQT file. The protein structures of α-glucosidase (PDB ID: 5ZCE) and PTP1B (PDB ID: 5QG3) are from the RCSB database ( https: / / www.rcsb.org / ), the protein structure was treated with water molecules, ligands, hydrogenation, etc. on the Pymol and Autodock platforms, and the protein was energy minimized and the geometry optimized and exported as a PDBQT file. The molecular docking processing and optimization 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 fully wrapped around the protein, and then molecular docking was performed. In addition, the complex of protein and small molecule was visualized and analyzed using Pymol.

[0049] Docking results Molecular docking results Table 4. Docking results of compound 1 and target protein Target protein Compound Binding energy (kcal / mol) α-Glucosidase 1 -8.5 PTP1B 1 -21.4 Analysis of compound-protein interactions In this experiment, compound 1 was combined with α -glucosidase and PTP1B target protein were molecularly docked. The molecular docking results showed that the compound had a good binding effect with the target protein and the matching degree was high ( Fig.11 and Fig.12 ), the binding energy is less than -6 kcal / mol. The complex formed by the docking compound and the protein was visualized using Pymol2.1 software to obtain the binding mode of the compound and the protein. According to the binding mode, the amino acid residues that bind to the compound and the protein pocket can be clearly seen. Compound 1 and α - The PHE-144 amino acid in the active site of the glucosidase protein forms hydrophobic interactions and multiple hydrogen bond interactions with ASP-60, ILE-143, GLN-256, THR-409, and ARG-411 amino acids; Compound 1 forms multiple hydrophobic interactions with amino acids TYR-46 and VAL-49 in the active site of the PTP1B protein, and multiple hydrogen bond interactions with amino acids TYR-46, ASP-48, ASP-181, and GLN-266; it has strong binding ability and plays an important role in anchoring small molecules in the protein pocket; The above specific implementation method part specifically introduces the analytical method involved in the present invention. It should be noted that the above introduction is only to help those skilled in the art better understand the method and ideas of the present invention, rather than limiting the relevant content. Without departing from the principle of the present invention, those skilled in the art may also make appropriate adjustments or modifications to the present invention, and the above adjustments and modifications shall also fall within the scope of protection of the present invention.

Claims

1. A glycoside compound from the root of Herba Cynanchifoliae, characterized in that: The structural formula is as follows: 。 2. The method for separating and extracting glycoside compounds from the root of Herba Cynanchifoliae according to claim 1, characterized in that: The steps include: A1. Solvent extraction: The root of Huazelan is crushed and extracted with ethanol, petroleum ether, ethyl acetate and n-butanol in sequence, and the n-butanol extract is concentrated to obtain extract a; B1. Extract a macroporous adsorption resin impurity removal: extract a is added with deionized water for ultrasonic dissolution, macroporous adsorption resin is soaked in deionized water and loaded onto the column, rinsed, wet loaded, left to stand overnight for adsorption, eluted with ethanol / deionized water system, and the eluate is concentrated under reduced pressure to obtain extract b; C1. Dissolve the extract b in methanol, and use reversed-phase silica gel column chromatography to crudely separate the extract. Use wet column packing, dry sample loading, and use a methanol / deionized water system for gradient elution to collect the eluate. D1, taking the eluate and subjecting it to HW-40F gel column chromatography to obtain a separated product; E1. The separated product was subjected to preliminary separation by reverse silica gel column chromatography, and the obtained fragments were prepared by semi-preparative HPLC to obtain compound 1.

3. The method for separating and extracting glycoside compounds from the root of Herba Cynanchifoliae according to claim 2, characterized in that: In step C1, 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 are used for gradient elution, and the eluate in the 30% methanol / 70% water portion, or the eluate in the 40% methanol / 60% water portion, or the eluate in the 60% methanol / 40% water portion is collected.

4. The method for separating and extracting glycoside compounds from the root of Herba Cynanchifoliae according to claim 2, characterized in that: The eluate in step D1 was separated by gel column chromatography HW-40F with methanol-water at a volume fraction of 10-25: 70-85 to obtain a separated product.

5. The method for separating and extracting glycoside compounds from the root of Herba Cynanchifoliae according to claim 2, characterized in that: In step E1, the separated product is separated by reverse silica gel column chromatography with methanol-water at 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.

6. A compound for inhibiting the activity of α-glucosidase or PTP1B, characterized in that: The compound includes the compound according to claim 1.

7. A drug for treating diabetes and obesity, characterized in that: The drug comprises the compound according to claim 1, and the drug is prepared for treating diabetes and obesity by inhibiting the activity of α-glucosidase or PTP1B.

8. The compound or drug according to claim 6 or 7, characterized in that The compound according to claim 1 is α - The amino acids in the active site of the glucosidase protein combine to form a site containing hydrophobic interactions and hydrogen bond interactions; The compound described in claim 1 combines with the amino acids in the active site of the PTP1B protein to form a site containing hydrophobic interactions and hydrogen bond interactions.

9. The compound or drug according to any one of claims 6 to 8, characterized in that The compound or medicament also includes a pharmaceutically acceptable excipient.

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