Dihydrochalcone glycoside compound and lipid-lowering application thereof
By extracting dihydrochalkol glycoside compounds from Russian tea, building a high-fat model to verify their lipid-lowering effect, solving the problem of large side effects and limited efficacy of existing lipid-lowering drugs, achieving high-efficiency and low-toxic lipid-lowering effects, and being suitable for the preparation of drugs and functional foods.
Patent Information
- Application Number
- CN202510688240.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-27
- Publication Date
- 2025-08-12
AI Technical Summary
Existing lipid-lowering drugs have great side effects and limited efficacy in treating dyslipidemia, especially in some patients, and it is necessary to develop safer and more effective lipid-lowering products.
Dihydrochalkol glycoside compounds were isolated from Russian tea, and a high-lipid model was constructed by oleic acid induced by HepG2 cells to construct a high-lipid model, verifying that it has a significant inhibitory effect on the accumulation of triglycerides in human liver cancer cells at low concentrations, and prepared into drugs or functional foods to play a lipid-lowering effect.
Dihydrochalkol glycoside compounds significantly reduce the number of lipid droplets in human liver cancer cells at low concentrations, and are better than the positive drug fenofibrate, providing a highly efficient and low-toxic lipid-lowering pathway. The raw materials are easy to obtain and are simple to prepare and industrialized.
Smart Images
Figure CN120463758A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a dihydrochalcone glycoside compound and application of the dihydrochalcone glycoside compound in the preparation of lipid-lowering products. Background Art
[0002] Dyslipidemia, characterized by elevated blood lipid levels, is a significant risk factor for cardiovascular disease, including coronary heart disease and stroke. It is often associated with metabolic disorders such as obesity, diabetes, and insulin resistance. Dysregulation of lipid metabolism can lead to the accumulation of triglycerides and cholesterol in the blood, which in turn promotes the development of atherosclerosis and subsequent cardiovascular complications.
[0003] The management of dyslipidemia typically involves lifestyle changes, including diet and exercise, and, when necessary, pharmacological intervention. Although a variety of lipid-lowering drugs, such as statins and fibrates, are commercially available, the search for new therapeutic agents with fewer side effects and improved efficacy continues, particularly for patients who do not respond adequately to or cannot tolerate existing treatments.
[0004] Natural products have emerged as a promising source of novel therapeutic agents due to their potential health benefits and low toxicity. Among them, polyphenols have attracted considerable attention due to their antioxidant, anti-inflammatory, and lipid-lowering properties. These compounds can modulate key enzymes and receptors in lipid metabolism, such as HMG-CoA reductase and peroxisome proliferator-activated receptors (PPARs), thereby reducing the synthesis and absorption of cholesterol and triglycerides.
[0005] O's tea is a kind of tea with the leaves of Begonia variegata ( Malus toringoides (Rehd.) Hughes) leaves, meticulously crafted using traditional techniques, this healthy tea boasts a long history of consumption in Tibet. Its sweet, slightly bitter flavor and cooling properties are associated with heat-clearing and detoxifying effects, lowering blood pressure and lipids, and providing antioxidant benefits. It has traditionally been used to treat hypertension, hyperlipidemia, and cardiovascular disease, and these medicinal benefits are documented in the book "Compendium of Tibetan Medicine." Modern phytochemical research reveals that it primarily contains flavonoids and phenolic acids, while modern pharmacological studies demonstrate its antioxidant, blood sugar-lowering, anti-inflammatory, immune-enhancing, and lipid-lowering activities. The discovery of compounds with lipid-lowering activity in this ethnic tea is of great significance. Summary of the Invention
[0006] The present invention provides a dihydrochalcone glycoside compound, which is separated from tea leaves and has the following chemical structure: .
[0007] Another object of the present invention is to use the above-mentioned dihydrochalcone glycoside compound in the preparation of lipid-lowering products.
[0008] The lipid-lowering product of the present invention comprises a dihydrochalcone glycoside compound as its ingredient (or active ingredient), and may further comprise one or more pharmaceutically acceptable excipients or functional food excipients, or may be combined with other active ingredients to exert therapeutic effects. The product may be prepared into pharmaceutical or functional food tablets, pills, powders, capsules, granules, etc., and may also be prepared into various forms such as oral liquids.
[0009] Compared with the prior art, the advantages and technical effects of the present invention are: 1. The present invention isolates a dihydrochalcone glycoside compound from tea leaves, and uses oleic acid to induce HepG2 cells to obtain a high-fat model. The high-fat model is treated with the dihydrochalcone glycoside compound. The experimental results show that the dihydrochalcone glycoside compound can effectively downregulate the accumulation of TG in human liver cancer cells (HepG2) treated with oleic acid at low concentrations and reduce the number of lipid droplets. The effect is better than that of the positive drug fenofibrate. The results show that the dihydrochalcone glycoside compound has a lipid-lowering effect; 2. The raw materials for obtaining the dihydrochalcone glycoside compound of the present invention are readily available, the preparation method is simple and rapid, the purity is high, and it is easy to industrialize and produce; the present invention provides a new approach for developing highly effective and low-toxic lipid-lowering products. BRIEF DESCRIPTION OF THE DRAWINGS
[0010] Figure 1 The results of the cytotoxicity test of dihydrochalcone glycoside compounds on HepG2 cells are shown; Figure 2 The results of the effect of dihydrochalcone glycoside compounds on TG accumulation in HepG2 cells induced by oleic acid (OA) are shown in the figure. ## p < 0.01 for comparison between OA group and blank group; ** p < 0.01 for comparison between sample group and OA group; Figure 3 It is the result of the effect of dihydrochalcone glycoside compounds on lipid droplets in HepG2 cells induced by oleic acid (OA). DETAILED DESCRIPTION
[0011] The present invention will be further described below with reference to the examples. However, the scope of the present invention is not limited to the following examples. Those skilled in the art will appreciate that various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention. The instruments, reagents, and materials involved in the following examples, unless otherwise specified, are conventional instruments, reagents, and materials already available in the prior art and can be obtained through regular commercial channels. The experimental methods, detection methods, etc. involved in the following examples, unless otherwise specified, are conventional experimental methods and detection methods already available in the prior art.
[0012] Example 1: Extraction and separation of dihydrochalcone glycoside compounds 1. After pulverizing 10 kg of Ouse tea with a powder grinder, ultrasonically extract the mixture three times with 80% methanol aqueous solution for 30 min each time. The combined extracts were collected and concentrated using a rotary evaporator at 55°C to obtain 1.2 kg of Ouse tea extract. 2. Dissolve the Russian tea extract in water and extract with ethyl acetate five times. Collect and combine the ethyl acetate phases and remove the solvent by rotary evaporation at 50°C to obtain 260 g of crude extract. 3. The crude extract was chromatographed on a silica gel column using dichloromethane-methanol (100:1-1:1) as the eluent. Thin layer chromatography (TLC) was used for detection. The eluates containing the same fractions were combined and the solvent was removed by rotary evaporation at 50°C to obtain seven components, Fr.1-Fr.7. 4. Component Fr.5 (9 g) was separated and purified by medium-pressure preparative chromatography using a gradient elution solution of 10%, 30%, 50%, 70% and 90% methanol in water. The eluate was collected and rotary evaporated to obtain five components (Fr. AE). Fr. B (800 mg) was separated on a silica gel column and eluted with chloroform and acetone (15:1, 10:1 and 5:1) to obtain three components (Fr. ac). Fr. b (120 mg) was then further separated and purified by preparative liquid chromatography to obtain a dihydrochalcone glycoside compound (9.3 mg).
[0013] The structural identification of the dihydrochalcone glycoside compound is as follows: Phloretin-2'-xyloside, yellow powder, HR-ESI-MS m / z : 405.39 [MH] - ;Molecular formula: C 20 H 22 O9. 1 H NMR (400 MHz, DMSO) δ H : 7.06 (2H, d, J = 8.2 Hz, H-2′, 6′), 6.69(2H, d, J = 8.2 Hz, H-3′, 5′), 6.18 (1H, d, J = 1.5 Hz, H-6), 5.96 (1H, d, J = 1.4 Hz, H-8), 3.44 (2H, m, H-2), 2.86 (2H, m, H-3); 13 C NMR (125 MHz, DMSO) δ C: 204.67 (s, C-4), 165.35 (s, C-7), 164.39 (s, C-9), 160.53 (s, C-5), 155.29 (s, C-4′), 131.49 (s, C-1′), 129.15 (d, C-3′, 5′), 114.99 (d, C-2′,6′), 105.26 (d, C-10), 101.22 (d, C-1′′), 96.90 (d, C-8), 94.20 (d, C-6),76.41 (d, C-3′′), 72.98 (d, C-4′′), 69.21 (d, C-2′′), 65.82 (t, C-5′′), 44.99 (t, C-2), 29.02 (t, C-3).
[0014] Example 2: Effect of dihydrochalcone glycoside compounds on triglyceride (TG) accumulation and oil droplet formation in oleic acid-treated human hepatoma cells (HepG2) A high-fat model was established by inducing HepG2 cells with oleic acid (OA). This stimulation causes lipid deposition in HepG2 cells, mimicking the pathogenesis of fatty liver disease caused by lipid metabolism disorders and resulting in the accumulation of large amounts of triglycerides (TG). A triglyceride (TG) assay kit was used to measure TG levels in the control and experimental groups to assess the inhibitory effect of the compounds on TG accumulation.
[0015] 1. Cell culture Human hepatic cell lines (HepG2, purchased from the Kunming Institute of Zoology, Chinese Academy of Sciences) were used for the experiments. The cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (Gibco, Grand Island, NY, USA) in an incubator at 37°C and 5% CO ; 2. Cytotoxicity evaluation 200 μL of HepG2 cells were seeded into 96-well cell culture plates (2×10 4cells), incubated at 37°C for 24 h, then 200 μL of different concentrations of dihydrochalcone glycoside compounds (10-100 μmol / L) were added to a 96-well plate. After incubation for 24 h, the culture medium was removed, and 200 μL of MTT (0.5 mg / mL) solution dissolved in PBS was added to the 96-well cell culture plate. The plate was placed in a 37°C, 5% CO2 incubator for further culturing for 4 hours, and then 200 μL of DMSO was added for 10 min. The absorbance was measured at 490 nm using a microplate reader, and the cell viability was calculated. The toxicity of dihydrochalcone glycoside compounds on HepG2 cells was evaluated by MTT assay. Cell viability (%) = (A 样品 / A 空白 )×100%, where A 样品 is the absorbance value of the experimental group with the addition of dihydrochalcone glycoside compounds, A 空白 is the absorbance value of the blank control group without adding dihydrochalcone glycoside compound; See the results Figure 1 ,The results in the figure show that dihydrochalcone glycoside compounds showed no toxicity to HepG2 cells within the concentration range of 10-100 μmol / L; 3. Evaluation of triglyceride (TG) accumulation HepG2 cells in the logarithmic growth phase were seeded in 6-well cell culture plates (3×10 5 cells, 2 mL), incubated at 37°C for 12 h, then washed with PBS, added with complete medium containing 0.4 mM oleic acid, and continued to culture at 37°C for 24 h. Then, different concentrations of dihydrochalcone glycoside compounds (10, 20, 40, 80, 100 μM) and fenofibrate (FNB, 40 μM) were added to the culture medium, and incubated at 37°C, 5% CO2 for 24 h. The accumulation of triglyceride (TG) in the cells was determined using a triglyceride (TG) content detection kit; See the results Figure 2 The dihydrochalcone glycoside compound had a significant inhibitory effect on the accumulation of TG at low concentrations (80μM and 100μM), and the effect was better than that of the positive drug fenofibrate, especially at 100μM, it showed good inhibitory activity.
[0016] 4. Oil Red O staining HepG2 cells (2 mL, concentration 3 × 10 5HepG2 cells were seeded at 100 μg / mL in 6-well plates and cultured at 37°C for 24 hours. Subsequently, 2.0 mL of complete medium containing 0.4 mM oleic acid was added to the 6-well plates and cultured at 37°C for another 24 hours. The cells were then treated with 2.0 mL of complete medium containing 40 μM dihydrochalcone glycosides and 40 μM fenofibrate and cultured for an additional 24 hours. HepG2 cells were stained with Oil Red O staining solution according to the kit instructions. Finally, cells were observed and photographed at 200x magnification using an Olympus microscope. See the results Figure 3 After HepG2 cells were treated with 400 μmol / L oleic acid, Oil Red O staining revealed dense accumulation of lipid droplets within the cells, indicating successful establishment of a lipid deposition model. In contrast, treatment with a dihydrochalcone glycoside compound (40 μM) reduced the number of lipid droplets to varying degrees, demonstrating an inhibitory effect on lipid deposition, even comparable to that of the positive control drug fenofibrate. These experimental results demonstrate the potent lipid-lowering activity of the compounds described herein, providing a new avenue for the development and utilization of oleic tea and the treatment of non-alcoholic fatty liver disease.
[0017] The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present invention without departing from the content of the technical solution of the present invention shall still fall within the scope of the technical solution of the present invention.
Claims
1. A dihydrochalcone glycoside compound having the following chemical structural formula: 。 2. Use of the dihydrochalcone glycoside compound according to claim 1 in the preparation of lipid-lowering products.