Dihydroisocumarin compound, preparation method and application thereof

Abstract

The present application relates to the preparation method and application of a dihydroisocumarin compound (3R, 4R)-3-(beta-D-glucopyranosylmethyl)-3, 4-dihydro-5, 6, 7-trihydroxy-4-(3'-methoxy-4'-hydroxyphenyl)-1H-[2]-benzopyran-1-one in Cornus officinalis, which can effectively solve the medication problem of diabetes, and the solution is that the chemical structure of the dihydroisocumarin compound is as follows: The dihydroisocumarin compound in the present application can have a certain improvement effect on insulin-induced HepG2 cell damage, can be used as a drug for treating diabetes, and the drug composition can be a monomer or a derivative thereof, which is an innovation of the dihydroisocumarin compound in Cornus officinalis and its application.

Description

I. Technical Field

[0001] This invention relates to the pharmaceutical field, and in particular to a dihydroisocoumarin compound extracted from Cornus officinalis, its preparation method, and its application. II. Background Technology

[0002] Diabetes mellitus is a chronic metabolic disease characterized by elevated blood sugar levels due to insulin deficiency (type 1 diabetes) or insulin resistance (type 2 diabetes). According to the International Diabetes Federation (IDF), approximately 537 million people aged 20-79 worldwide had diabetes in 2021, and this number is projected to rise to 783 million by 2045. Diabetes has a significant impact on human health, potentially leading to a variety of complications affecting multiple organ systems. These complications include cardiovascular disease, kidney disease, retinopathy, neuropathy, and diabetic foot ulcers, posing a serious threat to quality of life and lifespan. Current pharmacological interventions for diabetes management primarily include insulin therapy and oral hypoglycemic agents such as metformin, sulfonylureas, and thiazolidinediones. However, these treatments have limitations. Insulin therapy may require injections and has poor adherence, while oral medications may cause gastrointestinal adverse reactions and carry risks of hypoglycemia and weight gain. Therefore, there is an urgent need to develop innovative antidiabetic drugs to improve treatment outcomes and enhance patient well-being.

[0003] Cornus officinalis fruit, a traditional Chinese medicine, is widely used to treat various ailments, including dizziness and tinnitus, lower back and knee weakness, impotence, seminal emission, enuresis, frequent urination, and internal heat and thirst. Chemical studies have shown that Cornus officinalis fruit contains iridoid glycosides, tannins, flavonoids, and small amounts of volatile oils, organic acids, and triterpenoids. Pharmacological studies have demonstrated its various therapeutic effects, including hypoglycemic, antioxidant, anti-inflammatory, antitumor, neuroprotective, and hepatoprotective properties. Many pharmacological studies have focused on the antidiabetic effects of Cornus officinalis. For example, Han and He's research showed that oral administration of Cornus officinalis extract (300 mg / kg every 2 days, 400 mg / kg daily) and isolated loganin, morroniside, and ursolic acid (200 mg / kg daily) significantly reduced fasting blood glucose levels in diabetic rats and alleviated symptoms such as polydipsia, polyphagia, polyuria, and weight loss caused by diabetes (Y. Han, HW Jung, YK Park, Selective therapeutic effect of Cornus officinalis fruits on the damage of different organs in STZ-induced diabetic rats, Am J Chin Med 42 (2014) 1169-82; K. He, S. Song, Z. Zou, M. Feng, D. Wang, Y. Wang, X. Li, X. Ye, The hypoglycemic and synergistic effect of loganin, morroniside, and ursolic acid isolated from the fruits of Cornus officinalis, Phytother Res 30(2016)283-91.). Research by Peng et al. indicates that certain secoidoid dimers and their bioogenetic inhibitors from the fruits of Cornus officinalis may have potential α-glucosidase inhibitory activity (ZCPeng, J.He, XGPan, J.Zhang, YMWang, XSYe, CYXia, WWLian, Y.Yan, XLHe, WKZhang, JKXu, Secoidoid dimers and their bioogenetic inhibitors from the fruits of Cornus officinalis with potential therapeutic effects on type 2 diabetes, Bioorg Chem 117(2021)105399.).The anti-DM effects of Cornus officinalis extract and its effective components are clear, but there are no studies on the anti-DM activity of dihydroisocoumarin in Cornus officinalis. III. Summary of the Invention

[0004] In view of the above situation and to overcome the shortcomings of the prior art, the purpose of this invention is to provide a dihydroisocoumarin compound extracted from Cornus officinalis, its preparation method and application, which can effectively solve the medication problem of type 2 diabetes.

[0005] The technical solution provided by this invention is a dihydroisocoumarin compound extracted from Cornus officinalis, (3R,4R)-3-(β-D-pyranoseloxymethyl)-3,4-dihydro-5,6,7-trihydroxy-4-(3'-methoxy-4'-hydroxyphenyl)-1H-[2]-benzopyran-1-one (compound 1), with the following chemical structural formula:

[0006]

[0007] Its preparation method is as follows:

[0008] 1) Extract 10-100 kg of Cornus officinalis pulp by refluxing with 2-10 times the amount of deionized water at 80-100℃ 1-3 times. Concentrate the extract under reduced pressure to form an extract paste. Dissolve the extract in water and pass it through a D-101 macroporous adsorption resin column, eluting with ultrapure water and 95% ethanol respectively.

[0009] 2) The 95% ethanol eluent was concentrated under low temperature and reduced pressure to form a paste. After dissolving in water, it was extracted 3-10 times sequentially with petroleum ether, ethyl acetate, and n-butanol. The n-butanol extract was concentrated under pressure to form a paste, dissolved in water, and passed through a Diaion HP-20 macroporous adsorption resin column. It was eluted with an ethanol-water gradient (0:100, 20:80, 30:70, 40:60, 95:5; v / v) to obtain 5 fractions (BF1-BF5).

[0010] 3) Pass the BF2 fraction obtained in step 2) through an MCI column and elute with methanol-water (0:100, 20:80, 40:60, 60:40, 100:0; v / v) to obtain 5 fractions (BF2a-BF2e). Pass the BF2a fraction through a Sephadex LH-20 column and elute with a methanol-water gradient (20:80, 50:50, 70:30, 100:0; v / v) to obtain 5 fractions (BF2a1-BF2a5).

[0011] 4) Pass the BF2a4 obtained in step 3) through a Sephadex LH-2 column and elute with methanol-water (4:6-8:2; v / v) to obtain 9 fractions (BF2a4a-BF2a4i). Pass the BF2a4c through a silica gel column and elute with dichloromethane-methanol (12:1-5:1; v / v) to obtain 5 fractions (BF2a4c1-BF2a4c5).

[0012] 5) Pass the BF2a4c3 fraction obtained in step 4) through a silica gel column and elute with dichloromethane-methanol (10:1, v / v) to obtain 6 fractions (BF2a4c3a-BF2a4c3f). Pass the BF2a4c3d fraction through a silica gel column and elute with dichloromethane-methanol (10:1-0:1, v / v) to obtain 5 fractions (BF2a4c3d1-BF2a4c3d5).

[0013] 6) BF2a4c3d4 was purified by semi-preparative HPLC (9% methanol, 3 mL / min) using a COSMOSIL C18-MS-II column (5 μm, 10 × 250 mm), with 19% MeCN as the mobile phase and a flow rate of 3 mL / min. Compound 1 (t) was obtained. R =39.7min).

[0014] The application of the dihydroisocoumarin compounds described in this invention in the preparation of drugs for treating type 2 diabetes.

[0015] The present invention also provides a pharmaceutical composition comprising an active ingredient and a pharmaceutically acceptable carrier, wherein the active ingredient comprises the dihydroisocoumarin compound of claim 1 or a pharmaceutically acceptable salt, tautomer, stereoisomer, solvate, prodrug molecule, metabolite or mixture thereof.

[0016] The dihydroisocoumarin compound (3R,4R)-3-(β-D-glucopyranoylmethyl)-3,4-dihydro-5,6,7-trihydroxy-4-(3'-methoxy-4'-hydroxyphenyl)-1H-[2]-benzopyran-1-one described in this invention is obtained from Cornus officinalis pulp through solvent extraction, macroporous adsorption resin chromatography, silica gel column chromatography, gel column chromatography, MCI column chromatography, Sephadex LH-20 gel column chromatography, and preparative liquid phase separation. The preparation method is simple and easy to implement, and the product has high purity. Pharmacological activity studies show that this compound can significantly increase glucose consumption in an insulin-induced HepG2 cell insulin resistance model and significantly promote glucose uptake in the HepG2 cell insulin resistance model, indicating that this compound has potential therapeutic effects on diabetes and is of great significance in the development of antidiabetic drugs. IV. Description of the attached drawings

[0017] Figure 1 The dihydroisocoumarin compound 1 of this invention 1 H-NMR spectrum.

[0018] Figure 2 The dihydroisocoumarin compound 1 of this invention 13 C-NMR spectrum.

[0019] Figure 3 This is the DEPT-135 spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0020] Figure 4 The dihydroisocoumarin compound 1 of this invention 1 H- 1 H COSY spectrum.

[0021] Figure 5 This is the HSQC spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0022] Figure 6 This is the HMBC spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0023] Figure 7 This is the NOESY spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0024] Figure 8 This is the UV spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0025] Figure 9 The IR spectrum of compound 1, a dihydroisocoumarin compound of the present invention, is shown.

[0026] Figure 10 This is the HRESIMS spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0027] Figure 11 This is the CD spectrum of compound 1, a dihydroisocoumarin compound of the present invention.

[0028] Figure 12 This is a bar graph showing the effect of dihydroisocoumarin compound 1 of the present invention on the viability of HepG2 cells.

[0029] Figure 13 This is a line graph showing the effect of dihydroisocoumarin compound 1 of the present invention on glucose consumption in HepG2 cells.

[0030] Figure 14 This is a flow cytometry image of glucose uptake in HepG2 cells by dihydroisocoumarin compound 1 of the present invention.

[0031] Figure 15This is a fluorescence image of the expression of insulin receptor INSR in HepG2 cells by dihydroisocoumarin compound 1 of the present invention.

[0032] Figure 16 This is the structural diagram of a dihydroisocoumarin compound 1 of the present invention. V. Detailed Implementation Methods

[0033] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings.

[0034] Example 1:

[0035] In a specific implementation of the present invention, a method for preparing dihydroisocoumarin includes the following steps:

[0036] 1) 10 kg of Cornus officinalis pulp was extracted twice by reflux at 90°C with 3 times the amount of deionized water. The extract was concentrated under reduced pressure to form a paste. The paste was dissolved in water and passed through a D-101 macroporous adsorption resin column, and eluted with ultrapure water and 95% ethanol, respectively.

[0037] 2) The 95% ethanol eluent was concentrated under low temperature and reduced pressure to form a paste. After dissolving in water, it was extracted six times sequentially with petroleum ether, ethyl acetate, and n-butanol. The n-butanol extract was concentrated under pressure to form a paste, dissolved in water, and passed through a Diaion HP-20 macroporous adsorption resin column. It was eluted with an ethanol-water gradient (0:100, 20:80, 30:70, 40:60, 95:5; v / v) to obtain five fractions (BF1-BF5).

[0038] 3) Pass the BF2 fraction obtained in step 2) through an MCI column and elute with methanol-water (0:100, 20:80, 40:60, 60:40, 100:0; v / v) to obtain 5 fractions (BF2a-BF2e). Pass the BF2a fraction through a Sephadex LH-20 column and elute with a methanol-water gradient (20:80, 50:50, 70:30, 100:0; v / v) to obtain 5 fractions (BF2a1-BF2a5).

[0039] 4) Pass the BF2a4 obtained in step 3) through a Sephadex LH-2 column and elute with methanol-water (4:6-8:2; v / v) to obtain 9 fractions (BF2a4a-BF2a4i). Pass the BF2a4c through a silica gel column and elute with dichloromethane-methanol (12:1-5:1; v / v) to obtain 5 fractions (BF2a4c1-BF2a4c5).

[0040] 5) Pass the BF2a4c3 fraction obtained in step 4) through a silica gel column and elute with dichloromethane-methanol (10:1, v / v) to obtain 6 fractions (BF2a4c3a-BF2a4c3f). Pass the BF2a4c3d fraction through a silica gel column and elute with dichloromethane-methanol (10:1-0:1, v / v) to obtain 5 fractions (BF2a4c3d1-BF2a4c3d5).

[0041] 6) BF2a4c3d4 was purified by semi-preparative HPLC (9% methanol, 3 mL / min) using a COSMOSIL C18-MS-II column (5 μm, 10 × 250 mm), with 19% MeCN as the mobile phase and a flow rate of 3 mL / min. Compound 1 (t) was obtained. R =39.7min).

[0042] Example 2:

[0043] In a specific implementation of the present invention, a method for preparing dihydroisocoumarin includes the following steps:

[0044] 1) 10 kg of Cornus officinalis pulp was extracted three times by reflux with three times the amount of deionized water at 90°C. The extract was concentrated under reduced pressure to form a paste. The paste was dissolved in water and passed through a D-101 macroporous adsorption resin column, and eluted with ultrapure water and 95% ethanol, respectively.

[0045] 2) The 95% ethanol eluent was concentrated under low temperature and reduced pressure to form a paste. After dissolving in water, it was extracted four times sequentially with petroleum ether, ethyl acetate, and n-butanol. The n-butanol extract was concentrated under pressure to form a paste, dissolved in water, and passed through a Diaion HP-20 macroporous adsorption resin column. Elution was performed using an ethanol-water gradient (0:100, 20:80, 30:70, 40:60, 95:5; v / v) to obtain five fractions (BF1-BF5).

[0046] 3) Pass the BF2 fraction obtained in step 2) through an MCI column and elute with methanol-water (0:100, 20:80, 40:60, 60:40, 100:0; v / v) to obtain 5 fractions (BF2a-BF2e). Pass the BF2a fraction through a Sephadex LH-20 column and elute with a methanol-water gradient (20:80, 50:50, 70:30, 100:0; v / v) to obtain 5 fractions (BF2a1-BF2a5).

[0047] 4) Pass the BF2a4 obtained in step 3) through a Sephadex LH-2 column and elute with methanol-water (4:6-8:2; v / v) to obtain 9 fractions (BF2a4a-BF2a4i). Pass the BF2a4c through a silica gel column and elute with dichloromethane-methanol (12:1-5:1; v / v) to obtain 5 fractions (BF2a4c1-BF2a4c5).

[0048] 5) Pass the BF2a4c3 fraction obtained in step 4) through a silica gel column and elute with dichloromethane-methanol (10:1, v / v) to obtain 6 fractions (BF2a4c3a-BF2a4c3f). Pass the BF2a4c3d fraction through a silica gel column and elute with dichloromethane-methanol (10:1-0:1, v / v) to obtain 5 fractions (BF2a4c3d1-BF2a4c3d5).

[0049] 6) BF2a4c3d4 was purified by semi-preparative HPLC (9% methanol, 3 mL / min) using a COSMOSIL C18-MS-II column (5 μm, 10 × 250 mm), with 19% MeCN as the mobile phase and a flow rate of 3 mL / min. Compound 1 (t) was obtained. R =39.7min).

[0050] Example 3:

[0051] In a specific implementation of the present invention, a method for preparing dihydroisocoumarin includes the following steps:

[0052] 1) 10 kg of Cornus officinalis pulp was extracted three times by reflux with an equal amount of deionized water at 90°C. The extract was concentrated under reduced pressure to form a paste. The paste was dissolved in water and passed through a D-101 macroporous adsorption resin column, and eluted with ultrapure water and 95% ethanol, respectively.

[0053] 2) The 95% ethanol eluent was concentrated under low temperature and reduced pressure to form a paste. After dissolving in water, it was extracted eight times sequentially with petroleum ether, ethyl acetate, and n-butanol. The n-butanol extract was concentrated under pressure to form a paste, dissolved in water, and passed through a Diaion HP-20 macroporous adsorption resin column. Elution was performed using an ethanol-water gradient (0:100, 20:80, 30:70, 40:60, 95:5; v / v) to obtain five fractions (BF1-BF5).

[0054] 3) Pass the BF2 fraction obtained in step 2) through an MCI column and elute with methanol-water (0:100, 20:80, 40:60, 60:40, 100:0; v / v) to obtain 5 fractions (BF2a-BF2e). Pass the BF2a fraction through a Sephadex LH-20 column and elute with a methanol-water gradient (20:80, 50:50, 70:30, 100:0; v / v) to obtain 5 fractions (BF2a1-BF2a5).

[0055] 4) Pass the BF2a4 obtained in step 3) through a Sephadex LH-2 column and elute with methanol-water (4:6-8:2; v / v) to obtain 9 fractions (BF2a4a-BF2a4i). Pass the BF2a4c through a silica gel column and elute with dichloromethane-methanol (12:1-5:1; v / v) to obtain 5 fractions (BF2a4c1-BF2a4c5).

[0056] 5) Pass the BF2a4c3 fraction obtained in step 4) through a silica gel column and elute with dichloromethane-methanol (10:1, v / v) to obtain 6 fractions (BF2a4c3a-BF2a4c3f). Pass the BF2a4c3d fraction through a silica gel column and elute with dichloromethane-methanol (10:1-0:1, v / v) to obtain 5 fractions (BF2a4c3d1-BF2a4c3d5).

[0057] 6) BF2a4c3d4 was purified by semi-preparative HPLC (9% methanol, 3 mL / min) using a COSMOSIL C18-MS-II column (5 μm, 10 × 250 mm), with 19% MeCN as the mobile phase and a flow rate of 3 mL / min. Compound 1 (t) was obtained. R =39.7min).

[0058] The pharmacological activity results of the dihydroisocoumarin compounds prepared in this invention show that they can significantly increase glucose consumption in an insulin-induced HepG2 cell insulin resistance model, and can be used as a drug for the treatment of type 2 diabetes. Relevant experimental data are as follows:

[0059] I. Structural identification of the compounds of this invention:

[0060] Compound 1 is an amorphous powder (MeOH). Its molecular formula was determined to be C0.05 by HRESIMS. 23 H 26 O 13 (m / z533.1286,[M+Na] +Its infrared spectrum shows the presence of hydroxyl (3457 cm⁻¹) and carbonyl (1682 cm⁻¹). Its NMR data are very similar to (3S,4S)³-(β-D-glucopyranosyloxymethyl)-3,4-dihydro-5,6,7-trihydro-xy-4-(4′-hydroxy-3′-methoxyphenyl)-1H-[2]-benzopyran-1-one [AMAbdulmagid,VNLaurence,G.Moroy,C.Moretti,C.Lavaud,Dihydroisocoumarin glucosides fromstem bark of Caryocar glabrum,Phytochemistry 68(2007)2439-2443.], which indicates that compound 1 may have the same skeleton as the previously reported compound, which was verified by 2DNMR spectroscopy. 1 H NMR, 13 CNMR, 1 H- 1 H COSY, HSQC, and HMBC spectra were used to determine its planar chemical structure. Chiral HPLC analysis confirmed that the attached sugar group was D-glucose, and further analysis was conducted using... 3 J H-1″,H-2″ (7.7 Hz) determined that its glucoside group was β-configuration. Its NOESY spectrum showed the correlation between H-3a / H-4, H-1” / H-4, H-3 / H-2', and H-3 / H-6', indicating that H-3 and 3-methoxy-4-hydroxyphenyl groups were located on the same side of the lactone ring (α orientation), while H-4 and β-D-pyranoseloxymethyl groups were located on the other side (β orientation). The absolute configuration of the compound was determined by comparing experimental ECD spectra and calculated ECD spectra. Therefore, the structure of compound 2 was determined to be (3R,4R)-3-(β-D-pyranoseloxymethyl)-3,4-dihydro-5,6,7-trihydroxy-4-(3'-methoxy-4'-hydroxyphenyl)-1H-[2]-benzopyran-1-one.

[0061] Compound 1 1 H-NMR and 13 The C-NMR spectral data are shown in Table 1.

[0062] Table 1. Compound 1 1 H NMR (500MHz, CD3OD) and 13 C NMR spectral data

[0063]

[0064]

[0065] *no = This signal was not observed.

[0066] II. Antidiabetic effects of the dihydroisocoumarin compounds of this invention

[0067] 1) CCK-8 assay to determine the effect of compound 1 on the cell viability of HepG2 cells

[0068] HepG2 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum at 37°C in a 5% CO2 incubator. Cells were seeded in 96-well plates at a density of 1 × 10⁶ cells / well. 5 Cells / well. Preparation of insulin resistance (IR): HepG2 cells were starved in serum-free DMEM for 24 h, cultured for 24 h, then the medium was changed to serum-free DMEM containing 20 μM insulin, and incubated for another 24 h. Then, DMEM medium, metformin medium (10 μM), or medium containing monomeric compound (10.0 μM) were added, and incubated for 24 h. The medium was changed, 100 μL of fresh serum-free medium was added, followed by 10 μL of CCK-8 reagent, and incubated at 37℃ for 1 h. The optical density (OD) value of each well was measured at 450 nm using a microplate spectrophotometer. Results are shown below. Figure 12 Compound 1 had no significant effect on the cell viability of HepG2 cells at doses of 0.3125, 0.625, 1.25, 2.5, 5.0, 10.0, and 20.0 μM.

[0069] 2) Determination of relative glucose consumption

[0070] HepG2 cells (1×10⁵ cells / well) cultured in 96-well plates were treated with insulin and compound 1. The culture medium was then collected, and glucose concentration was measured using a glucose assay kit. Glucose consumption (GC) was obtained by subtracting the glucose content of the experimental group's culture medium from the glucose content of the original DMEM medium. The relative glucose consumption (RGC) was calculated using the formula: RGC = GC / OD.

[0071] See results Figure 13 Compound 1 significantly increased the relative glucose consumption in the HepG2 insulin resistance model in a dose-dependent manner, with an optimal concentration of 20.0 μM.

[0072] 3) Detection of glucose uptake capacity using the 2-NBDG probe method

[0073] HepG2 cells were cultured in 6-well plates (1×10⁻⁶ cells / well). 5Cells were treated with insulin and compound 1 (1 mL / well) and heated for 24 h with 10.0 μM insulin for 24 h, as described above. Then, the culture medium was gently poured out, and 1.0 mL of DMEM medium was added to each well, along with a glucose uptake fluorescent probe (2-NBDG, 20.0 μM). The cells were incubated in a cell culture incubator for 1 h. Cells were collected and transferred to flow cytometry tubes, washed twice with PBS, and resuspended in 500 μL of DRAQ5 (1:500) nuclear dye. Cell fluorescence intensity was detected using Flow Sight.

[0074] See results Figure 14 The fluorescence of cells in the compound 1 group was significantly enhanced, suggesting that compound 1 can significantly improve the glucose uptake capacity of the HepG2 insulin resistance model.

[0075] 4) Detection of insulin receptor (INSR) expression by cellular immunofluorescence

[0076] Cells at 1×10 5 Cell Carrier was seeded at a density of 100 μL / well. TM -96 plates were prepared, and compound 1 was administered at a dose of 10.0 μM, following the same procedure as in 1). After 24 h of culture, the plates were fixed with 4% paraformaldehyde (room temperature, 20 min). They were then blocked with 10% goat serum (room temperature, 90 min), followed by incubation overnight at 4°C with INSR (1:500) and Tubulin (1:1000), and then goat anti-rabbit IgG H&L (Alexa) was added. 488) and goat anti-mouse IgG H&L (Alexa) 594) was pre-adsorbed with secondary antibody (1:1000), incubated at 37℃ for 1 h, and then the cell nuclei were re-stained with DAPI. Fluorescence intensity was measured using a high-content cell imaging system with excitation wavelengths of 488 nm and 597 nm. The relative expression levels of the protein were normalized using Harmony 4.8. The results are shown below. Figure 15 The fluorescence of cells in the compound 1 group was significantly enhanced, suggesting that compound 1 can significantly increase the number of insulin receptors (INSR) in the cell membrane of the HepG2 insulin resistance model and promote glucose transport and utilization.

[0077] This invention utilizes the pulp of Cornus officinalis, a plant belonging to the genus Cornus in the family Cornaceae. Through solvent extraction, macroporous adsorption resin column chromatography, MCI gel, Sephadex LH-20 column chromatography, silica gel column chromatography, and preparative high-performance liquid chromatography, a novel dihydroisocoumarin compound is obtained. Pharmacological activity results show that this compound can significantly increase glucose consumption in an insulin-induced HepG2 cell insulin resistance model. By increasing the number of insulin receptors (INSRs) on the cell membrane in the HepG2 insulin resistance model, it enhances cellular glucose uptake and consumption, making it suitable for use as a treatment for type 2 diabetes. This invention represents an innovation in dihydroisocoumarins and their applications, possessing significant application and promotional value.

Claims

1. A dihydroisocoumarin compound extracted from Cornus officinalis, characterized in that, For (3) R 4 R )-3-( β -D-pyranoglucosylmethyl)-3,4-dihydro-5,6,7-trihydroxy-4-(3'-methoxy-4'-hydroxyphenyl)-1 H -[2]-Benzopyran-1-one (1), chemical structural formula is: 。 2. The method for preparing a dihydroisocoumarin compound extracted from Cornus officinalis according to claim 1, characterized in that, Includes the following steps: 1) Extract 10-100kg of Cornus officinalis pulp by refluxing with 2-10 times the amount of deionized water at 80-100℃ 1-3 times. Concentrate the extract under reduced pressure to form an extract paste. Dissolve the extract in water and pass it through a D-101 macroporous adsorption resin column, and elute with ultrapure water and 95% ethanol respectively. 2) The 95% ethanol eluent was concentrated into a paste under low temperature and reduced pressure. After being dissolved in water, it was extracted 3-10 times sequentially with petroleum ether, ethyl acetate and n-butanol. The n-butanol extract was concentrated into a paste under pressure and dissolved in water. It was then passed through a Diaion HP-20 macroporous adsorption resin column and eluted with gradients of ethanol-water at volume ratios of 0:100, 20:80, 30:70, 40:60 and 95:5 to obtain 5 fractions. 3) Pass the second fraction obtained in step 2) through an MCI column and elute with a methanol-water gradient of volume ratios of 0:100, 20:80, 40:60, 60:40, and 100:0 to obtain 5 fractions; pass the first fraction through a Sephadex LH-20 column and elute with a methanol-water gradient of volume ratios of 20:80, 50:50, 70:30, and 100:0 to obtain 5 fractions. 4) Pass the fourth fraction obtained in step 3) through a Sephadex LH-2 column and elute with methanol-water at a volume ratio of 4:6-8:2 to obtain 9 fractions; pass the third fraction through a silica gel column and elute with dichloromethane-methanol at a volume ratio of 12:1-5:1 to obtain 5 fractions. 5) Pass the third part obtained in step 4) through a silica gel column and elute with dichloromethane-methanol at a volume ratio of 10:1 to obtain 6 parts; pass the fourth part through a silica gel column and elute with dichloromethane-methanol at a volume ratio of 10:1-0:1 to obtain 5 parts; 6) The fourth part obtained in step 5) was purified by semi-preparative HPLC. The chromatography column was a COSMOSIL C18-MS-II column with a specification of 5 μm and a diameter of 10 × 250 mm. The mobile phase was 9% methanol, and the flow rate was 3 mL / min. Compound 1 was obtained, with a retention time t. R =39.7min.

3. The use of the dihydroisocoumarin compound of claim 1 in the preparation of a drug for treating type 2 diabetes.

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