Preparation method and application of a new benzofuran component from Huazelan

Abstract

The present invention discloses a preparation method and application of a new benzofuran component in Huazelan. The compound is prepared from Huazelan ( Eupatorium chinense ) was isolated from the extracted part. The root extract of Huazelan was purified and separated by extraction method, macroporous adsorption resin for impurity removal, HW-40F gel column chromatography and high performance liquid chromatography to obtain a new benzofuran compound. The inhibitory activity of the above-isolated compound on the diabetes-related targets α-glucosidase and protein tyrosine phosphatase 1B (PTP1B) was tested. The experimental results showed that the compound had good inhibitory activity on the two enzymes. Molecular docking experiments also showed that the compound had good binding energy with the two target proteins α-glucosidase and protein tyrosine phosphatase 1B. Therefore, the compound can be used to prepare drugs for preventing, delaying or treating diseases mediated by α-glucosidase or PTP1B, especially type 2 diabetes and obesity, or as a lead compound for such drugs.

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 benzofuran compounds from the roots of Huazelan, and specifically to a separation, purification and preparation method of a new benzofuran compound from the roots of Huazelan, as well as its inhibitory activity on α-glucosidase and protein tyrosine phosphatase 1B (PTP1B). Background Art

[0002] Huazelan is a perennial herbaceous plant of the genus Eupatorium in the Asteraceae family. Its dried rhizomes are used as a folk medicine for the treatment of diphtheria, sore throat, and other throat diseases. It is known as the "holy medicine for throat diseases." It is a commonly used Chinese herbal medicine in western Hubei, the Tujia ethnic group in western Hunan, and Guangdong Province. Its chemical constituents mainly include terpenes, flavonoids, and benzofurans. Studies have shown that it has pharmacological activities such as anti-tumor, protein tyrosine phosphatase (PTP1B) inhibition, anti-inflammatory, antibacterial, antiviral, and hypoglycemic effects (Li Degang, Ke Yinqian, Zou Kun, et al. Study on the anti-inflammatory activity of chemical constituents from the petroleum ether fraction of Huazelan rhizomes. Chinese Pharmacology and Clinics. 2022, 38(03): 106-110.).

[0003] α-glucosidase, present in the small intestinal mucosa, is one of the primary target enzymes for controlling postprandial blood glucose. It converts carbohydrates in food into glucose, reversibly competing with sugar molecules for enzyme binding sites on the intestinal brush border epithelial cells to delay the formation of monosaccharides, thereby achieving balanced blood glucose levels. Currently, α-glucosidase inhibitors are one of the first-line drugs for the clinical treatment of type 2 diabetes.

[0004] The protein-tyrosine phosphatases (PTPs) family is a class of receptor-like cytoplasmic signal transduction enzymes that dephosphorylate the tyrosine residues of their substrates (Kim et al ., Int. J. Mol. Sci. 2018, 19: 2708), thereby regulating metabolic processes in various cells and maintaining body homeostasis. Protein tyrosine phosphatase 1B (PTP1B) is a typical non-transmembrane tyrosine phosphatase and a negative transducer of insulin signaling. Insulin resistance, especially in adipose tissue and muscle, is closely associated with the development of type 2 diabetes mellitus (T2DM) (Eur. J. Pharmacol. 2005, 519: 182-190; Kim et al., 2014). et al., J. Clin. Invest. 2000, 105: 1791-1797). Furthermore, PTP1B overexpression has been detected in studies of breast, ovarian, and prostate cancers, and at all stages of tumor development (Mol. Cell. Biol. 1998, 18, 2965-2975; J. Natl. Cancer Inst. 1996, 86, 372-378), indicating that PTP1B is closely involved in many important tumorigenic signaling pathways. Therefore, PTP1B inhibitors could be used to treat or prevent cancer, or to slow its progression once it develops. Simultaneous inhibition of both α-glucosidase and PTP1B can lower postprandial blood glucose levels while enhancing insulin sensitivity and reducing pancreatic β-cell load, ultimately maintaining stable blood glucose levels. Furthermore, by increasing leptin levels, PTP1B can reduce body weight in obese diabetic patients.

[0005] Natural products, particularly those derived from plants, have long been a crucial source for the discovery of innovative drugs with novel structures and mechanisms of action (J. Nat. Prod. 2020, 83: 770-803). Plant-derived active ingredients are characterized by diverse skeletons, novel structures, multiple targets, and low toxicity and side effects. They are often selected and evolved through natural processes, capable of effectively binding to biomacromolecules and exhibiting excellent activity. Therefore, screening and discovering novel, highly effective α-glucosidase and PTP1B inhibitors from plant-derived active ingredients holds significant research value. Summary of the Invention

[0006] The first object of the present invention is to provide a benzofuran compound from the root of Herba Cynanchifoliae and a preparation method thereof. The second object is to provide a use of the compound in the preparation of a drug for preventing or treating diseases mediated by α-glucosidase or protein tyrosine phosphatase PTP1B.

[0007] The compound having the ability to inhibit α-glucosidase or PTP1B activity has the following structural formula:

[0008]

[0009] Eupbenzofuran A, compound 1.

[0010] The technical solution of the present invention is to use a benzofuran compound extracted from the root of Huazelan in the preparation of drugs for preventing, delaying or treating diseases mediated by α-glucosidase or protein tyrosine phosphatase PTP1B.

[0011] The preparation method of the Huazelan acetylene compound comprises the following steps:

[0012] A1. Solvent extraction: The root of Herba Cynanchifoliae was crushed and extracted with ethanol, petroleum ether, ethyl acetate, and n-butanol in sequence. The n-butanol extract was concentrated to obtain extract a.

[0013] B1. Extract a impurity removal by macroporous adsorption resin: Extract a was dissolved in deionized water by ultrasonication. Macroporous adsorption resin (D101) was soaked in deionized water and loaded onto the column. The column was rinsed, wet-loaded, and allowed to stand overnight for adsorption. The column was eluted with an ethanol / deionized water system. The eluate was concentrated under reduced pressure to obtain extract b.

[0014] C1. Dissolve the extract b in methanol and perform a crude fractionation of the extract by reverse-phase silica gel column chromatography. Use wet column packing and dry loading, perform gradient elution using a methanol / deionized water system, and collect the eluate.

[0015] D1, taking the eluate and performing preliminary separation on a reverse silica gel column chromatography to obtain the separated product;

[0016] E1. The separated product was separated by HW-40F gel column chromatography, and the obtained fragments were prepared by semi-preparative HPLC to obtain compound 1.

[0017] In step C1, gradient elution was 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. The eluate from the 60% methanol / 40% water portion, the eluate from the 80% methanol / 20% water portion, or the eluate from the 100% methanol portion was collected.

[0018] The eluate in step D1 was sequentially separated by reverse silica gel column chromatography with 10% methanol / water, 20% methanol / water, 30% methanol / water, 40% methanol / water, 50% methanol / water, 60% methanol / water, 70% methanol / water, 80% methanol / water, 90% methanol / water, and 100% methanol to collect the separated products of 90% methanol / water and / or 100% methanol.

[0019] In step E1, the separated product is separated by HW-40F gel column chromatography using methanol-water at a volume fraction of 60-75:25-40;

[0020] In the semi-preparative HPLC, the mobile phase is acetonitrile-water 60:40, and the flow rate is 1-2 mL / min.

[0021] A medicine for treating diabetes and obesity, comprising the prepared compound 1.

[0022] A compound for inhibiting alpha-glucosidase or PTP1B activity, comprising the prepared compound 1.

[0023] A drug for preventing, delaying or treating diabetes and obesity is prepared by inhibiting the activity of α-glucosidase or PTP1B, and the drug comprises the prepared compound 1.

[0024] The compound 1 is prepared by α -Glucosidase protein active site amino acids combine to form sites containing hydrophobic interactions and hydrogen bond interactions;

[0025] 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.

[0026] The compound or drug of the present invention further includes a pharmaceutically acceptable excipient.

[0027] Compared with the prior art, this application has at least one of the following beneficial effects:

[0028] 1. Compound 1 of the present application has significant α-glucosidase or PTP1B inhibitory activity. There are no public reports on the compound's α-glucosidase or PTP1B inhibitory activity and its use in related diseases.

[0029] 2. For the first time, a compound isolated from the n-butanol extract of the root of Huazelanthus chinensis has been found to inhibit α-glucosidase or PTP1B activity. This compound has broad application prospects for the development of drugs to prevent or treat diseases mediated by α-glucosidase or PTP1B, such as diabetes and obesity. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Figure 1 is the chemical structure of compound 1.

[0031] Figure 2 For compound 1 1 H-NMR.

[0032] Figure 3 For compound 1 13 C-NMR.

[0033] Figure 4 This is the DEPT135 spectrum of compound 1.

[0034] Figure 5 is the HSQC spectrum of compound 1.

[0035] Figure 6 For compound 1 1 H-1 H COSY spectrum.

[0036] Figure 7 is the HMBC spectrum of compound 1.

[0037] Figure 8 is the NOESY spectrum of compound 1.

[0038] Figure 9 This is the experimental ECD spectrum of compound 1.

[0039] Figure 10 This is the ECD spectrum calculated by TD-DFT theory for compound 1.

[0040] Figure 11 Interaction analysis diagram of compound 1 and α-glucosidase protein (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).

[0041] Figure 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

[0042] The following examples are used to further illustrate the present invention. Obviously, the examples described are only part of the embodiments of this application, not all of them. Based on the examples in this application, all other examples obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0043] Example 1: Separation and purification of a benzofuran compound extracted from the roots of Herba Cynanchifoliae

[0044] The preparation method comprises the following steps:

[0045] Step A, solvent extraction: The roots of Huazelan were dried and crushed. Extracted three times with 95% ethanol under reflux at room temperature. After concentration on a rotary evaporator, the extract was dispersed in water and extracted three times with petroleum ether, ethyl acetate, and n-butanol. The n-butanol extract was concentrated to obtain extract a.

[0046] Step B, impurity removal of extract a using a macroporous adsorption resin: extract a was taken, 300 g of deionized water was added and ultrasonically dissolved, 1000 g of macroporous adsorption resin (type D101) was soaked in deionized water for 6 h and then loaded onto a column, and rinsed with deionized water for at least three column volumes. The solvent was lowered to about 2 cm above the packing surface, and the sample was loaded using a wet method. The sample was allowed to stand overnight for adsorption, and eluted with 50% ethanol / 50% deionized water. The eluate was rinsed until it was colorless, and the eluate was concentrated under reduced pressure to obtain extract b.

[0047] Step C, column chromatography crude fractionation: Take the above extract b, dissolve it with a small amount of methanol, add it to 90 g of reverse-phase silica gel (200-300 mesh), and mix the sample; weigh 1200 g of 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 approximately 2 cm higher than the packing surface, use dry method to load the sample, after loading, plug it with an appropriate amount of absorbent cotton as a protective layer, and combine the results of high performance liquid chromatography 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, with each gradient washing for 2 column volumes to obtain 8 fractions (Fr.AH).

[0048] Thin layer chromatography, high performance liquid chromatography and HPLC-DAD scanning analysis showed that the 60% methanol / 40% water portion Fr.F was the component where the target compound was located.

[0049] Step D: The 60% methanol / 40% water fraction Fr.F was subjected to high-performance liquid chromatography (HPLC) analysis (analysis conditions: 10% methanol / water to 100% methanol, 30 minutes, flow rate 1 mL / min, wavelength 203 nm). After preliminary separation by reverse-phase silica gel column chromatography (10% methanol / water to 100% methanol), 10 fractions Fr.F (1 to 10) were obtained.

[0050] The reversed-phase silica gel column chromatography employed the following conditions: 10% methanol / water, 20% methanol / water, 30% methanol / water, 40% methanol / water, 50% methanol / water, 60% methanol / water, 70% methanol / water, 80% methanol / water, 90% methanol / water, and 100% methanol. HPLC-DAD scanning analysis indicated that Fr.F-9 was the target compound.

[0051] Step E: Fr. F-9 was initially separated by HW-40F gel column chromatography (methanol-water 75:25, volume ratio) to obtain four fragments (Fr. F9-1 ​​to F9-4). Compound 1 (5.9 mg) was obtained from Fr. F-9-3 by semi-preparative HPLC (acetonitrile-water 60:40, 2 mL / min, YMC C18 column).

[0052] The NMR spectroscopy data and mass spectrometry data of compound 1 are as follows:

[0053] Compound 1: Colorless oily liquid. Optical rotation data [α] 25 D +8.6° (c 0.10, methanol), UV (methanol) λ max =239, 350 nm; by high resolution mass spectrometry HR-ESI-MS m / z : 313.1059 [M+Na] + (C 16 H 18 NaO5, calcd.313.1052), its molecular formula is determined to be C 16 H 18 O5, unsaturation degree is 8. 1 The H-NMR spectrum (Table 1) shows two olefin proton signals. δ H 8.31 (H-4) and 7.07 (H-7), the compound has 3 methyl groups ( δ H 2.72 (H-16), 1.50 (H-17) and 1.45 (H-14). 13 The C-NMR spectrum and DEPT135 spectrum showed 16 carbon signals, including 3 methyl carbon signals. δ C 27.9 (C-16), 27.9 (C-17), and 27.2 (C-14); 2 methylene carbon signals δ C 37.69 (C-12) and δ C 36.77 (C-11); 2 oxygen-linked carbon signals δ C 68.74 (C-10) and 65.69 (C-13); 1 carbonyl carbon signal δ C 205.05 (C-15); 8 olefin carbon signals δ C160.04 (C-6), 158.92 (C-8), 156.9 (C-2), 124.9 (C-4), 120.7 (C-5), 119.7 (C-9), 117.22 (C-3), 99.52 (C-7).

[0054] Table 1. Comparison of Compound 1 and Reference Compounds 1 H-NMR and 13 C-NMR data comparison (solvent DMSO- d 6)

[0055]

[0056] In the HMBC spectrum, it can be observed that δ H 1.50 (H-17), δ H 1.73 (H-11), respectively and δ C There is a correlation at 65.69 (C-10), confirming the presence of a hydroxyl group and a methyl group at the C-10 position. In the HMBC spectrum, there is a correlation between H-14 and C-13, C-12, and C-2, and between H-16 and C-15 and C-5; 1 H- 1 In the H COSY spectrum, there is a correlation between H-11 and H-12. Thus, the planar structure of compound 1 was obtained ( Figure 1 ).

[0057] In the NOESY spectrum ( Figure 8 ) observed in H-17 / H-11 β 、H-14 / H-11 β The correlation between H-14 and C-17 indicates that H-14 and C-17 are on the same side of the C ring. Finally, the relative configuration of compound 1 is determined. Its absolute configuration is calculated by TD-DFT theory ECD spectrum ( Figure 12 ) and experimental ECD spectrum ( Figure 11 ) (positive cotton effect at 242 nm and positive cotton effect at 360 nm) compared and determined to be 10 S、 13 R， After searching, it was found to be a new compound and named Eupbenzofuran A.

[0058] Example 2: Inhibitory activity test of compound 1 on α-glucosidase and PTP1B

[0059] Experimental methods

[0060] α-glucosidase activity assay

[0061] 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-pyranoside 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 UV absorption at 405 nm. Acarbose was used as a positive control. The amount of product after adding the sample was detected to determine the inhibition rate of the sample. 80 μL of sample (n-butanol extract of Huazelan or compound 1) solution (PBS buffer, pH 7.2) was added to the test wells of a 96-well plate, and 20 μL of α β-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 initiate the reaction. After another 15 min incubation, 80 μL of 1 mol / L Na₂CO₃ was added to terminate the reaction. To the blank control, 80 μL of 1% PBS was added instead of the sample solution. The absorbance (A) at 405 nm was used to quantify the amount of pNPG released. The following groups were set up: enzyme activity group (20 μL enzyme + 80 μL reaction buffer + 20 μL substrate), enzyme blank group (100 μL reaction buffer + 20 μL substrate), sample group (80 μL sample + 20 μL enzyme + 20 μL substrate), and sample blank group (80 μL sample + 20 μL reaction buffer + 20 μL substrate). Acarbose solution was used as a positive control. The inhibition rate was calculated as follows: Inhibition rate = 1 - (A sample - A sample blank) / (A enzyme activity - A enzyme blank).

[0062] PTP1B enzyme activity assay

[0063] The determination method of PTP1B was referred to the literature ( Funct. Foods, 2018, 41:232-239; Acta Pharmaceutica Sinica, 2019, 54(3): 510-513. 10 μL of sample (n-butanol extract of Huazelan or compound 1) was added to 170 μL of reaction buffer (50 mM citric acid (pH 7.4), 50 mM NaCl, 2 mM dithiothreitol (DTT), and 1.1 mM EDTA). 20 μL of recombinant PTP1B solution (1 mg / mL, 1 μL) was added to each well and mixed. The reaction mixture was preheated at 37°C for 15 min using a block heater. 10 μL of the reaction substrate p-nitrophenyl phosphate (pNPP) (33 mM) was added and reacted 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. According to the above steps, the enzyme activity group (20 μL of enzyme + 170 μL of reaction buffer solution + 10 μL of substrate), enzyme blank group (190 μL of reaction buffer + 10 μL of reaction substrate), sample group (170 μL of sample + 20 μL of enzyme + 10 μL of reaction substrate), and sample blank group (170 μL of sample + 20 μL of reaction buffer solution + 10 μL of reaction substrate) were set up, and sodium orthovanadate aqueous solution and oleanolic acid solution were used as positive controls.

[0064] The inhibition rate was calculated according to the following formula: Inhibition rate = 1-(A sample-A sample blank) / (A enzyme activity-A enzyme blank)

[0065] 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:

[0066] Table 2. Results of compound 1 inhibiting α-glucosidase activity

[0067]

[0068] The test results show that the above-mentioned benzofuran compounds from the roots of Huazelan have good α-glucosidase inhibitory activity.

[0069] Table 3. Results of compound 1 inhibiting PTP1B enzyme activity

[0070]

[0071] The test results show that the above-mentioned benzofuran compounds in the roots of Huazelan have certain PTP1B enzyme inhibitory activity.

[0072] Example 3: Docking of compound 1 with two protein target molecules

[0073] Compound 1 for this docking was constructed using ChemDraw, then imported into Chem3D software for optimization and energy minimization using the MM2 module. The result was saved as an sdf file as the ligand for molecular docking. The structure 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) were obtained from the RCSB database ( https: / / www.rcsb.org / The protein structure was processed using Pymol and Autodock platforms, including water molecule removal, ligand removal, and hydrogenation. Energy minimization and geometry optimization were performed, and the protein was exported as a PDBQT file. Molecular docking processing and optimization were performed using the Grid module in Autodock. The PDBQT file was imported into the software, fully encapsulated, and then molecular docking was performed. Complexes between the protein and small molecules were visualized and analyzed using Pymol.

[0074] Docking results

[0075] Molecular docking results

[0076] Table 4. Docking results of compound 1 and target protein

[0077]

[0078] Compound-protein interaction analysis

[0079] In this experiment, compound 1 was combined with α Molecular docking of the target protein of -glucosidase and PTP1B was performed. The molecular docking results showed that the compound had a good binding effect with the target protein and a high degree of match ( Figure 11 and Figure 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 amino acids PHE-163 and ALA-200 in the active site of the β-glucosidase protein form multiple hydrophobic interactions and multiple hydrogen bond interactions with amino acids GLN-256, ASP-327, GLN-328, and ARG-411.

[0080] Compound 1 forms multiple hydrophobic interactions with GLU-75, GLU-76, LEU-234, LYS-248, VAL-249, and GLU-252 amino acids in the active site of the PTP1B protein, and multiple hydrogen bond interactions with GLU-76, ALA-77, ARG-238, and VAL-249 amino acids; it has strong binding ability and plays an important role in anchoring small molecules in the protein pocket.

[0081] The above detailed description of the analytical methods involved in the present invention provides a detailed introduction. It should be noted that the above description is intended solely to help those skilled in the art better understand the methods and concepts of the present invention, and is not intended to limit the relevant content. Without departing from the principles of the present invention, those skilled in the art may make appropriate adjustments or modifications to the present invention, and such adjustments and modifications shall also fall within the scope of protection of the present invention.

Claims

1. Benzofuran compounds from the root of Herba Cynanchifoliae, characterized in that: The structural formula is as follows: Compound 1.

2. The method for separating and extracting benzofuran compounds from the root of Herba Cynanchifoliae according to claim 1, characterized in that: The steps include: A1. Solvent extraction: The root of Herba Cynanchifoliae was crushed and extracted with ethanol, petroleum ether, ethyl acetate, and n-butanol in sequence. The n-butanol extract was concentrated to obtain extract a. B1. Extract a impurity removal with macroporous adsorption resin: Extract a was dissolved in deionized water by ultrasonication. The macroporous adsorption resin was soaked in deionized water and loaded onto the column. The column was rinsed, wet-loaded, and allowed to stand overnight for adsorption. The column was eluted with an ethanol / deionized water system, and the eluate was concentrated under reduced pressure to obtain extract b. C1. Dissolve the extract b in methanol and perform a crude fractionation of the extract by reverse-phase silica gel column chromatography. Use wet column packing and dry loading, perform gradient elution using a methanol / deionized water system, and collect the eluate. D1, taking the eluate and performing preliminary separation on a reverse silica gel column chromatography to obtain the separated product; E1. The separated product was separated by HW-40F gel column chromatography, and the obtained fragments were prepared by semi-preparative HPLC to obtain compound 1.

3. The method for separating and extracting benzofuran compounds from the root of Herba Cynanchifoliae according to claim 2, wherein: In step C1, gradient elution was 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. The eluate from the 60% methanol / 40% water portion, the eluate from the 80% methanol / 20% water portion, or the eluate from the 100% methanol portion was collected.

4. The method for separating and extracting benzofuran compounds from the root of Herba Cynanchifoliae according to claim 2, wherein: The eluate in step D1 was sequentially separated by reverse silica gel column chromatography with 10% methanol / water, 20% methanol / water, 30% methanol / water, 40% methanol / water, 50% methanol / water, 60% methanol / water, 70% methanol / water, 80% methanol / water, 90% methanol / water, and 100% methanol to collect the separated products of 90% methanol / water and / or 100% methanol.

5. The method for separating and extracting benzofuran compounds from the root of Herba Cynanchifoliae according to claim 2, wherein: In step E1, the separated product is separated by HW-40F gel column chromatography using methanol-water at a volume fraction of 60-75:25-40; In the semi-preparative HPLC, the mobile phase is acetonitrile-water 60:40, and the flow rate is 1-2 mL / min.

6. A drug for treating diabetes and obesity, characterized in that: The drug comprises the compound 1 according to claim 1.

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

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

9. The compound or drug according to claim 7 or 8, characterized in that The compound 1 according to claim 1 is prepared by α -Glucosidase protein active site amino acids combine to form sites containing hydrophobic interactions and hydrogen bond interactions; The compound 1 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.

10. The compound or drug according to claim 9, characterized in that The compound or drug also includes a pharmaceutically acceptable excipient.

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