A polyphenol compound extracted from old fragrant yellow and application thereof in preparation of medicine for inhibiting enteritis

Abstract

This invention relates to a polyphenolic compound extracted from *Pterocarya stenoptera* and its application in the preparation of drugs for inhibiting enteritis. The polyphenolic compound is the first to be isolated from *Pterocarya stenoptera*, and is a novel, stable, and naturally occurring flavan-3-ol dimer monomer, rather than a polymer degradation product. The preparation method can stably produce high-purity polyphenolic compounds, suitable for the large-scale preparation of novel active ingredients from natural products. Experiments have shown that the polyphenolic compound of this invention possesses intestinal barrier repair capabilities, can inhibit inflammatory factors in the intestinal environment, and is more effective than commonly used clinical drugs. Furthermore, it is naturally derived, highly safe, and has significant clinical application value.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology for digestive system diseases, and in particular to a polyphenolic compound extracted from *Cinnamomum camphora* and its application in the preparation of drugs for inhibiting enteritis. Background Technology

[0002] Lao Xiang Huang (also known as Lao Xiang Yuan or Fo Shou Xiang Huang) is a unique preserved fruit made from the fruit of the bergamot (Citrus medica var. sarcodactylis), a plant in the Rutaceae family. It is produced through traditional processes including salting, rinsing, steaming, soaking in Chinese herbs, sun-drying, and long-term aging. A classic food and medicinal product of the Lingnan region, it is considered the foremost of the "Three Treasures of Chaozhou." The finished product has a dense texture and rich aroma. Traditionally used in folk medicine, it is believed to stimulate appetite, regulate qi, aid digestion, resolve phlegm, and promote saliva production. It is said to alleviate symptoms of upper gastrointestinal issues such as abdominal distension, belching, nausea, and stomach discomfort, thus possessing both nutritional and traditional health benefits.

[0003] In recent years, modern analytical techniques have been gradually applied to the study of the quality and composition of aged Xianghuang (a type of fermented rice wine). This has confirmed that the aging process is accompanied by dynamic changes in functional components such as volatile terpenes, total flavonoids, total phenols, and anthocyanins, and that significant differences exist in flavor compounds and physicochemical indicators across different aging years. Related research has largely focused on quality evaluation, age identification, volatile substance composition, and functional component content determination, providing preliminary basis for the quality grading and standardization of aged Xianghuang.

[0004] However, since traditional efficacy guidelines for the application of Lao Xiang Huang (a type of medicinal herb) have focused on aspects such as stagnation of Qi in the stomach and intestines and indigestion, modern research on Lao Xiang Huang has almost entirely concentrated on the upper digestive tract, quality evaluation, and component analysis. There has been no adaptive research on the entire digestive system or systematic research on key pharmacological pathways.

[0005] Therefore, it is necessary to break through the limitations of existing technology, systematically carry out research on the active ingredients and mechanisms of action of Lao Xiang Huang, fill the gaps in the field, and provide a basis for expanding its application scope and enhancing the scientific value of the product. Summary of the Invention

[0006] The purpose of this invention is to disclose a polyphenolic compound extracted from old sage and its application in the preparation of drugs for inhibiting enteritis, so as to solve one or more technical problems existing in the existing methods and provide at least one beneficial option or create conditions.

[0007] To achieve the above objectives, the present invention provides the following technical solution: A first aspect of this invention is to provide a polyphenolic compound. The structural formula of the polyphenolic compound is shown in Formula I: .

[0008] The polyphenolic compound is a novel flavan-3-ol dimer isolated for the first time from *Aster tataricus*. It exhibits a molecular weight (m / z 593), a clear NMR signal, regular mass spectrometry fragmentation, and a single chromatographic peak behavior. Furthermore, its structure contains a stable intramolecular ether-bonded dimer framework. These characteristics are fundamentally different from the random, unstable, and polydisperse degradation fragments produced by acid degradation of highly polymerized condensed tannins. Therefore, it can be proven that the polyphenolic compound is an independently existing, novel, stable, and naturally occurring flavan-3-ol dimer monomer, rather than a polymer degradation product.

[0009] A second aspect of the present invention is to provide a method for preparing the polyphenolic compounds described in the first aspect of the present invention. The method involves using aged sage as a raw material, employing accelerated solvent extraction to obtain a total extract; obtaining the active enriched fraction through organic solvent extraction; removing impurities by gel filtration chromatography; and finally separating the polyphenolic compounds using preparative normal-phase high-performance liquid chromatography.

[0010] In a further embodiment of the second aspect of the present invention, the specific steps of the accelerated solvent extraction include: placing aged huanghuasin in an accelerated solvent extractor, using anhydrous ethanol as the solvent, and extracting for 10-15 minutes at a temperature of 90-105 °C and a pressure of 10.0-10.5 MPa, with the extraction cycle repeated 2-5 times; the total extract is obtained after removing the solvent from the extracted product. By controlling the extraction temperature, pressure, time, and number of cycles, the polyphenolic components are more fully dissolved, significantly improving the extraction efficiency; at the same time, the oxidation and destruction of active ingredients caused by high-temperature and long-term treatment are avoided, maximizing the preservation of the anti-inflammatory activity of the compounds.

[0011] In a further embodiment of the second aspect of the present invention, the organic solvent extraction involves redissolving the total extract and then performing liquid-liquid extraction with dichloromethane at a volume ratio of 1:1, repeating this process 2 to 5 times; followed by freeze-drying to obtain the active enriched fraction. The total extract is redissolved in an appropriate amount of distilled water, and the subsequent addition of dichloromethane removes lipid-soluble impurities. Freeze-drying at -50 °C under vacuum allows the active enriched fraction to form a porous powder, which dissolves more quickly in subsequent processing, facilitating subsequent gel column and preparative normal-phase high-performance liquid chromatography loading, and improving separation efficiency.

[0012] In a further embodiment of the second aspect of the present invention, the impurity removal by gel filtration chromatography involves dissolving the active enriched fraction in methanol, filtering it, loading it onto a gel column for chromatography, eluting with methanol to remove impurities, and then eluting the target component with a methanol / acetone / water mixed solvent. The gel filtration chromatography may use a Sephadex series gel column (cross-linked dextran gel), preferably a 2.5 cm × 100 cm Sephadex LH-20 gel chromatography column, with a flow rate of 1.0 mL / min.

[0013] In a further embodiment of the second aspect of the present invention, the volume ratio of the methanol / acetone / water mixed solvent is 3:2:5. This ratio provides moderate elution strength and high selectivity, enabling precise elution of the target compound, reducing cross-components, improving the purity and yield of the target fraction, and ensuring the stability of monomer separation.

[0014] In a further embodiment of the second aspect of the present invention, the preparative normal-phase high-performance liquid chromatography uses a normal-phase glycol-based column with gradient elution in a hexane / ethyl acetate / methanol system, and a detection wavelength of 275-285 nm. The normal-phase glycol-based column and the hexane / ethyl acetate / methanol elution system provide high separation, good peak shape, and high recovery for polyphenolic compounds; 275-285 nm is the characteristic absorption wavelength of polyphenols, ensuring sensitive and stable detection, and allowing for the one-time acquisition of high-purity monomeric compounds with purity meeting the requirements for structural identification and pharmacological activity studies.

[0015] A third aspect of this invention involves providing application scenarios for the polyphenolic compounds described in the first aspect. Specifically, the polyphenolic compounds can be used to prepare drugs for preventing or treating enteritis and repairing intestinal mucosal barrier damage. Experiments have shown that the polyphenolic compounds can inhibit pro-inflammatory polarization of macrophage M1, promote anti-inflammatory polarization of M2, downregulate various inflammatory factors, and significantly upregulate the expression of tight junction proteins ZO-1 and Occludin, achieving a synergistic effect of anti-inflammation and intestinal barrier repair.

[0016] A fourth aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the polyphenolic compound described in the first aspect of the invention, and pharmaceutically acceptable excipients.

[0017] In a further embodiment of the fourth aspect of the present invention, the excipients include at least one selected from fillers, binders, disintegrants, lubricants, wetting agents, solubilizers, pH adjusters, preservatives, and antioxidants.

[0018] The present invention has the following advantages and effects compared with the prior art: 1. A novel polyphenolic compound was extracted from aged huanghuali wine. Unlike the structural fragments formed by polymer degradation in aged wine, this polyphenolic compound possesses an intramolecular ether-bonded dimer structure, existing as a stable monomer. It is speculated that this structure may have formed during the aging process of aged huanghuali wine through complex reaction pathways involving polyphenol oxidation, degradation, and condensation.

[0019] 2. The provided preparation method can stably produce high-purity polyphenolic compounds, which is suitable for the large-scale preparation of novel active ingredients from natural products.

[0020] 3. The polyphenolic compounds described in this invention have been experimentally proven to have intestinal barrier repair capabilities and can inhibit inflammatory factors in the intestinal environment. Their effects are superior to commonly used clinical drugs, and they are naturally derived, highly safe, and have significant clinical application value. Attached Figure Description

[0021] Figure 1 This is the carbon NMR spectrum from Example 2; Figure 2 This is the LC-ESI-MS / MS spectrum from Example 2; Figure 3 This is a bar chart showing the effect of polyphenolic compounds on TEER of Caco-2 monolayer cells under the action of macrophage inflammatory factors in Example 3. Figure 4 These are the results of the FITC-dextran permeability test in Example 3; Figure 5 This refers to the Western blot detection results in Example 3; Figure 6 This refers to the qRT-PCR detection results in Example 3; Figure 7 These are the ELISA test results from Example 4; Figure 8 This is the HPLC detection result of the old fragrant yellow control experiment in Example 6. Detailed Implementation

[0022] The following embodiments further illustrate the content of the present invention, but should not be construed as limiting the present invention. Any modifications and substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and essence of the present invention are within the scope of the present invention.

[0023] Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0024] Example 1: Preparation of the polyphenolic compounds The preparation process is as follows: (1) Weigh about 10.0 g of the old fragrant yellow sample and place it in the extraction cell of an accelerated solvent extractor (ASE, Thermo Dionex). Use anhydrous ethanol as solvent and perform static extraction for 10 minutes at a temperature of 100 ℃ and a pressure of 10.3 MPa, and repeat the process 3 times. Combine the extracts and remove the solvent by rotary evaporation under reduced pressure at 40 ℃ to obtain the total extract.

[0025] (2) The total extract was redissolved in an appropriate amount of distilled water and then subjected to liquid-liquid extraction with dichloromethane at a volume ratio of 1:1, repeated 3 times to remove lipid-soluble impurities. Subsequently, it was freeze-dried under vacuum at -50 °C to obtain the active enriched fraction.

[0026] (3) Take the active enriched fraction, dissolve it in 50% (v / v) methanol and filter it through a 0.45 μm filter membrane, then load it onto a 2.5 cm × 100 cm Sephadex LH-20 gel chromatography column. The elution process is as follows: first, elute with 50% (v / v) methanol at 1.0 mL / min to remove small molecule impurities; then elute with a methanol / acetone / water (30:20:50, v / v / v) mixed solvent and collect the target fraction.

[0027] (4) The target component was concentrated under reduced pressure and freeze-dried, and then further separated and purified by preparative normal-phase high-performance liquid chromatography. The chromatographic column was a normal-phase Diol preparative column, and the mobile phase was a hexane / ethyl acetate / methanol system for gradient elution. The detection wavelength was set to 280 nm, and the flow rate was set according to the specifications of the preparative column. The eluted components were collected in segments according to the ultraviolet absorption peaks, and after concentration and freeze-drying, polyphenolic compounds as shown in Formula I were obtained.

[0028] Example 2, Identification of the polyphenolic compounds The compounds obtained in Example 1 were analyzed by carbon nuclear magnetic resonance spectroscopy and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS / MS).

[0029] (1) The compound was dissolved in deuterated methanol, and the results of the carbon NMR spectroscopy analysis are as follows: Figure 1As shown, the chemical shifts of the carbon atom signals were observed to be δ = 157.3 ppm, 145.3 ppm, 137.1 ppm, 118.4 ppm, 116.2 ppm, 114.3 ppm, 110.1 ppm, 96.7 ppm, 78.2 ppm, and 60.1 ppm, respectively. These chemical shifts can be determined to originate from C7 / C5 / C8a (157.3), C3' / C4' (145.3), C1' (137.1), C6' (118.4), C5' (116.2), C2' (114.3), C6 (110.1), C8 / C4a (96.7), C2 (78.2), and C4 (60.1) on the flavan-3-ol skeleton. This indicates that the compound has a typical flavan-3-ol skeleton and belongs to the condensation tannin class. Notably, the C4 signal peak appears at 60.1 ppm, significantly higher than the C4 signals of other flavan-3-ols (typically 35–40 ppm). This indicates that oxygen substitution has occurred at the C4 position. Because oxygen is highly electronegative, it reduces the electron cloud density of the carbon atom, leading to a deshielding effect and thus increasing the chemical shift.

[0030] (2) The LC-ESI-MS / MS spectral results are as follows Figure 2 As shown. In positive ion mode, the characteristic peaks of the secondary mass spectrometry of this compound are m / z = 593, 575, 441, 425, 303, and 289. Among them, m / z = 593 is the molecular ion peak; m / z = 575 is generated by the dehydration of hydroxyl groups; m / z = 425 and 441 are characteristic fragments generated by the RDA cleavage of heterocycles; m / z = 303 is a gallocatechin fragment containing a trihydroxy B ring; and m / z = 289 is a catechin fragment.

[0031] Testing confirmed that the compound has a typical flavan-3-ol dimer skeleton. Carbon spectroscopy data showed that there was a characteristic oxygen substitution at the C4 position. The fragmentation pattern and molecular weight information of secondary mass spectrometry further confirmed that it is a novel flavan-3-ol polyphenol compound and not a degradation fragment of a known polymer.

[0032] Example 3: Verification of the intestinal barrier repair ability of the polyphenolic compounds Caco-2 intestinal cells were cultured in the upper layer of a transwell, and macrophages were seeded in the lower layer to establish an intestinal cell-macrophage co-culture model. The specific method is as follows: (1) Caco-2 cells were cultured in DMEM high-glucose medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. RAW264.7 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. All cells were cultured in a 37 ℃, 5% CO2 incubator. After culture, Caco-2 cells were seeded in the upper chamber of a Transwell chamber at a seeding density of 1×10⁻⁶ cells / mL. 5 For each cell / insert, add 200 μL of culture medium to the upper chamber and 600 μL to the lower chamber. Change the culture medium every 2 days during cell culture. Culture continuously for 14 days to allow Caco-2 to form a dense monolayer barrier. TEER (transepithelial resistance) values ​​are then measured. Cells are considered healthy when the TEER value is stable and greater than 350 Ω·cm. 2 RAW264.7 cells were then seeded in the lower chamber of the Transwell at a density of 2 × 10⁻⁶ cells / mL. 5 Cells / well. Subsequently, LPS was added to the lower chamber to a final concentration of 1 μg / mL to induce macrophages to release inflammatory factors, thereby disrupting the Caco-2 monolayer barrier. After 24 hours, once a significant decrease in TEER (denoted as TEER0) was recorded, 10 μmol / L of the polyphenolic compound involved in this invention was added. Cells without the polyphenolic compound served as a blank control, while cells with 10 μmol / L 5-aminosalicylic acid and 10 μmol / L dexamethasone served as positive controls. Cells were cultured for another 24 hours. TEER values ​​were measured before polyphenol treatment and at 8, 16, and 24 hours after treatment, and the relative changes in TEER were calculated.

[0033] Relative TEER (%) = 100 × TEER / TEER0 at different times after polyphenol treatment The results are as follows Figure 3 As shown, after treatment with polyphenolic compounds, the relative TEER value of Caco-2 cells gradually increased, reaching 179% at 24 hours; after treatment with the same dose of 5-aminosalicylic acid, the relative TEER value was 156%; and after treatment with dexamethasone, it was 126%. This result indicates that the polyphenolic compounds involved in this invention can significantly improve the barrier integrity of Caco-2 monolayer cells in the inflammatory microenvironment, with effects superior to commonly used positive controls 5-aminosalicylic acid and dexamethasone, demonstrating that this compound has a strong intestinal epithelial barrier repair capacity.

[0034] (2) After the transepithelial resistance test, the culture medium in the upper chamber was discarded, and the cell monolayer was gently washed with PBS. Then, FITC-dextran solution (4 kDa) was added to the upper chamber to a final concentration of 1 mg / mL. HBSS buffer was added to the lower chamber. After incubation at 37 °C in the dark for 1-2 hours, the liquid in the lower chamber was collected, and the fluorescence intensity was detected using a fluorescence microplate reader.

[0035] The results are as follows Figure 4 As shown, the FITC-dextran exudation from the Caco-2 cell layer after polyphenol treatment was significantly less than that in the blank control, indicating a significant reduction in the permeability of the Caco-2 cell layer and effective restoration of barrier integrity. Polyphenolic compounds can alleviate inflammation-induced epithelial barrier damage and inhibit intestinal hyperpermeability. Furthermore, the exudation of FITC-dextran was significantly less than that in the 5-aminosalicylic acid and dexamethasone treatment groups, indicating that this polyphenolic compound has a superior protective effect in reducing intestinal epithelial permeability and restoring tight junction structure. Its barrier repair effect is superior to the aforementioned commonly used anti-inflammatory drugs, further demonstrating its greater application potential in improving inflammation-related intestinal barrier dysfunction.

[0036] (3) After appropriate treatment, total protein was extracted from Caco-2 cells using RIPA lysis buffer (containing protease inhibitors). The cells were lysed on ice for 30 minutes and then centrifuged at 4 °C and 12,000 rpm for 10 minutes. The supernatant was collected. Protein concentration was determined using the BCA method: equal amounts of protein samples were separated by SDS-PAGE and transferred to a PVDF membrane. After blocking the PVDF membrane with 5% skim milk powder at room temperature, primary antibody ZO-1, Occludin, and internal control protein β-actin were added, and the membrane was incubated overnight at 4 °C. The next day, after washing, HRP-labeled secondary antibody was added, and the membrane was incubated at room temperature for 1 hour. After washing with TBST, the membrane was developed using ECL chemiluminescence reagent, and the signal was acquired using an imaging system to analyze the expression levels of each protein.

[0037] Western blot results are as follows Figure 5 As shown in the figure, compared with the healthy cell group, the expression of ZO-1 and Occludin in the model group Caco-2 cells was significantly reduced, suggesting that inflammatory stimulation disrupted the tight junction structure of the intestinal epithelium. Treatment with polyphenolic compounds significantly increased the expression of ZO-1 and Occludin proteins, and the recovery effect was better than that of the 5-aminosalicylic acid and dexamethasone treatment groups. These results indicate that the polyphenolic compounds involved in this invention can effectively reverse the inflammation-induced downregulation of tight junction proteins, thereby enhancing the integrity of the Caco-2 monolayer cell barrier.

[0038] (4) qRT-PCR results were used to evaluate the regulatory effect of polyphenolic compounds on macrophage polarization. M1 markers included iNOS and CD86, and M2 markers included Arg-1 and CD206.

[0039] The results are as follows Figure 6 As shown in the figure, compared with the healthy cell group, the expression of M1 markers iNOS and CD86 was significantly upregulated in the model group, while the expression of M2 markers Arg-1 and CD206 was significantly downregulated, suggesting that inflammatory stimulation induces macrophage polarization towards the pro-inflammatory M1 phenotype. After treatment with the polyphenolic compounds, the expression of iNOS and CD86 was significantly decreased, while the expression of Arg-1 and CD206 was significantly increased, with statistically significant differences (p<0.05), indicating that polyphenolic compounds can inhibit M1 polarization and promote M2 polarization.

[0040] In contrast, the 5-aminosalicylic acid and dexamethasone treatment groups also downregulated M1 markers and upregulated M2 markers to some extent, but the overall regulatory effect was weaker than that of the polyphenol treatment group. These results indicate that polyphenolic compounds can effectively regulate macrophage polarization, promoting their transformation from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype, thereby exerting an anti-inflammatory effect.

[0041] Example 4: The effect of the polyphenolic compounds on intestinal inflammatory factors. Similarly, after the treatment in Example 2, the supernatant from the upper and lower chambers of the Transwell culture was collected, centrifuged at 4 ℃ and 1000~3000 rpm for 10 minutes to remove cell debris, and the supernatant was used for subsequent detection.

[0042] Inflammatory factors were detected in the culture supernatants of the upper chamber (Caco-2 side) and the lower chamber (macrophage side) to differentiate between inflammatory responses originating from intestinal epithelial cells and immune cells. The upper chamber detected IL-8, IL-6, and MCP-1 levels to evaluate the inflammatory status of Caco-2 cells; the lower chamber primarily detected TNF-α and IL-1β levels to evaluate macrophage inflammatory responses. The levels of each inflammatory factor were measured using a commercially available enzyme-linked immunosorbent assay (ELISA) kit according to the manufacturer's instructions.

[0043] Test results as follows Figure 7As shown in the figure, compared with healthy cells, the levels of IL-6, IL-8, TNF-α, IL-1β, and MCP-1 in the model group were significantly increased, indicating that the inflammation model was successfully constructed. Specifically, the levels of TNF-α and IL-1β in the lower chamber were significantly increased, suggesting successful activation of macrophages; simultaneously, the levels of IL-8 and IL-6 in the upper chamber were increased, indicating that the inflammatory microenvironment induced the inflammatory response of Caco-2 cells. After treatment with polyphenolic compounds, the levels of inflammatory factors in both the upper and lower chambers were significantly reduced, with IL-6 decreasing from 185 pg / mL to 78 pg / mL, IL-8 from 240 pg / mL to 92 pg / mL, TNF-α from 165 pg / mL to 66 pg / mL, IL-1β from 138 pg / mL to 48 pg / mL, and MCP-1 from 210 pg / mL to 86 pg / mL, with statistically significant differences (p<0.05). In comparison, while 5-aminosalicylic acid and dexamethasone also reduced inflammatory cytokine levels, their overall effects were weaker than those of the polyphenol-treated group. These results indicate that the polyphenolic compounds involved in this invention can not only inhibit macrophage inflammatory responses but also alleviate inflammatory damage to intestinal epithelial cells, demonstrating good anti-inflammatory activity.

[0044] It inhibits the expression of iNOS, CD86, TNF-α, and IL-1β, while promoting their polarization towards the anti-inflammatory M2 and upregulating the expression of Arg-1 and CD206. It further upregulates the expression of intestinal epithelial tight junction proteins ZO-1 and Occludin and reduces the levels of inflammatory factors IL-6, IL-8, and MCP-1. Example 5: The regulatory effect of the polyphenolic compounds on macrophage polarization and intestinal epithelial tight junction proteins. Different concentrations of the polyphenolic compounds were used as samples (the amounts of the polyphenolic compounds in samples 1# to 9# were 10, 11, 12, 13, 14, 15, 16, 17, and 18 μmol / L, respectively). 5-Aminosalicylic acid at a concentration of 10 μmol / L and dexamethasone at a concentration of 10 μmol / L were used as control group 1 and control group 2, respectively, to treat intestinal cells / macrophages under inflammatory conditions. The various detection parameters of the treated cells are shown in Table 1.

[0045] Table 1

[0046] Table 1 shows that treatment with the polyphenolic compounds described in this invention significantly inhibited pro-inflammatory M1 polarization of macrophages, and significantly downregulated the expression of iNOS, CD86, TNF-α, and IL-1β. Simultaneously, treatment significantly promoted anti-inflammatory M2 polarization of macrophages, with marked upregulation of Arg-1 and CD206 expression. Furthermore, the expression of intestinal epithelial tight junction proteins ZO-1 and Occludin was significantly increased, while the levels of inflammatory factors such as IL-6, IL-8, and MCP-1 were significantly decreased, indicating that intestinal barrier function was effectively restored. These results confirm that the compounds of this invention can exert an inhibitory effect on enteritis by synergistically regulating macrophage polarization and intestinal epithelial barrier repair, and their effect is significantly superior to that of 5-aminosalicylic acid and dexamethasone.

[0047] Example 6: Effect of preparation time of aged huanghuali on the formation of polyphenolic compounds The preparation process of aged Xianghuang fruit includes fresh fruit pretreatment, salt fermentation, nine-times steaming and sun-drying, sugar soaking, and aging. To verify the formation time of the polyphenolic compounds, samples were taken from the preparation process up to the completion of the sugar soaking step as a control group; another group was aged Xianghuang fruit for one year as an experimental group. The polyphenolic compounds were extracted from both groups according to the method provided in Example 1. The HPLC detection results of the obtained products are as follows: Figure 8 As shown, Figure 8 The result shown at point A in the middle is the detection result of the experimental group. The characteristic peak of the polyphenolic compound appears at about 11 minutes. Figure 8 The result shown at point B is for the control group, which did not exhibit any obvious characteristic peaks. This further illustrates that the polyphenolic compounds are formed during the aging process. It will be apparent to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered illustrative and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of the equivalents of the claims are intended to be included within the present invention.

Claims

1. A polyphenolic compound, characterized in that, The structural formula is shown in Formula I: 。 2. The method for preparing the polyphenolic compound according to claim 1, characterized in that, Using old huanghuang as raw material, the total extract was obtained by accelerated solvent extraction; the active enriched fraction was obtained by organic solvent extraction; impurities were removed by gel filtration chromatography; and finally, the polyphenolic compounds were obtained by preparative normal-phase high-performance liquid chromatography.

3. The preparation method according to claim 2, characterized in that, The specific steps of the accelerated solvent extraction include: placing old huanghuang in an accelerated solvent extractor, using anhydrous ethanol as the solvent, and extracting for 10-15 minutes at a temperature of 90-105 ℃ and a pressure of 10.0-10.5 MPa, and repeating the extraction 2-5 times; after removing the solvent from the extract, the total extract is obtained.

4. The preparation method according to claim 3, characterized in that, The organic solvent extraction involves resuspending the total extract and then performing liquid-liquid extraction with dichloromethane at a volume ratio of 1:1, repeating the process 2 to 5 times; followed by freeze-drying to obtain the active enriched fraction.

5. The preparation method according to claim 4, characterized in that, The impurity removal by gel filtration chromatography involves dissolving and filtering the active enriched fraction with methanol, loading it onto a gel column for chromatography, eluting with methanol to remove impurities, and then eluting the target component with a methanol / acetone / water mixed solvent.

6. The preparation method according to claim 5, characterized in that, The volume ratio of the methanol / acetone / water mixed solvent is 3:2:

5.

7. The preparation method according to claim 5 or 6, characterized in that, The preparative normal-phase high-performance liquid chromatography uses a normal-phase diol-based column, with gradient elution using a hexane / ethyl acetate / methanol system, and a detection wavelength of 275~285 nm.

8. The use of the polyphenolic compound of claim 1 in the preparation of a medicament for the prevention or treatment of enteritis and the repair of intestinal mucosal barrier damage.

9. A pharmaceutical composition, characterized in that, The compound comprises a therapeutically effective amount of the polyphenolic compound of claim 1, and pharmaceutically acceptable excipients.

10. The pharmaceutical composition according to claim 9, characterized in that, The excipients include at least one of the following: fillers, binders, disintegrants, lubricants, wetting agents, solubilizers, pH adjusters, preservatives, and antioxidants.

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