A kind of Penthorum chinense Pursh compound, preparation method and application

Abstract

The present invention relates to the field of compounds of Penthorum chinense Pursh, and provides a compound of Penthorum chinense Pursh, a preparation method and an application, including crushing the Penthorum chinense Pursh medicinal materials, adding a 70% methanol-water mixture 10 times the amount of Penthorum chinense Pursh, heating and extracting 3 times at 50 °C, each time heating for 1.5 hours, filtering, concentrating the filtrate to 10 liters, and then adding one-fold water for dilution and filtration; performing C18 preparative crude fractionation, with a detection wavelength of 280 nm and a mobile phase of an acetonitrile-water system with 0.1% formic acid, including three segments, namely acetonitrile crude segment one, acetonitrile crude segment two, and acetonitrile crude segment three. Through research, the present invention finds that the new compound of Penthorum chinense Pursh, penthorin E, can have a good therapeutic effect on ethanol-induced hepatocyte diseases and reduce liver injury-related complications.

Description

Technical Field

[0001] The present invention relates to the field of compounds of Penthorum chinense Pursh, and more particularly to a compound of Penthorum chinense Pursh, a preparation method and an application thereof. Background Art

[0002] Penthorum chinense Pursh, scientifically named Penthorum chinense Pursh, was first recorded in "Herbal for Relief of Famine" in the Ming Dynasty and is a local Chinese medicinal material in Gulin County. It is mainly distributed in the Wumeng Mountain area at an altitude of about 1000 meters and has the effects of reducing jaundice and clearing damp-heat.

[0003] Ethanol, as a common raw material for beverages, often causes damage to liver cells and affects physical health after the beverages prepared with it are consumed.

[0004] Therefore, we propose a compound of Penthorum chinense Pursh, a preparation method and an application thereof and its preparation method to solve the above problems. Summary of the Invention

[0005] Technical Problems to be Solved

[0006] Aiming at the problems existing in the prior art, the purpose of the present invention is to provide a compound of Penthorum chinense Pursh, a preparation method and an application thereof, which solves the problem of ethanol-induced damage to liver cells.

[0007] Technical Solution

[0008] To solve the above problems, the present invention adopts the following technical solutions.

[0009] A compound of Penthorum chinense Pursh, including Figure 1 the structure shown.

[0010] A preparation method of the above compound of Penthorum chinense Pursh, including crushing Penthorum chinense Pursh medicinal materials, adding a 70% methanol-water mixture 10 times the amount of Penthorum chinense Pursh, heating and extracting 3 times at 50 °C for 1.5 hours each time, filtering, concentrating the filtrate to 10 liters, and then adding one-fold water for dilution and filtering;

[0011] C18 preparative crude fractionation, with a detection wavelength of 280 nm, and a mobile phase of an acetonitrile-water system with 0.1% formic acid, including three segments: acetonitrile crude segment one, acetonitrile crude segment two, and acetonitrile crude segment three;

[0012] The crude acetonitrile fraction two was concentrated to 1 L at 50 °C, filtered, and C18 was prepared again. The mobile phase was prepared with a 5% acetonitrile - water system containing 0.1% formic acid. The detection wavelength was 280 nm. The F4 target segment containing the target was collected, concentrated to a small volume at 50 °C, and repeatedly prepared and separated using a 14% methanol - water system and a 5% acetonitrile - water system. The qualified F4 and F4 - 2 solutions were collected, concentrated under reduced pressure at 50 °C, and then freeze - dried to obtain solid F4 and solid F4 - 2. Among them, F4 is xylocalyxin, and F4 - 2 is the above - mentioned penthorum chinense compound.

[0013] The crude acetonitrile fraction 1: Concentrated to 2 L at 50 °C, filtered, and separated again by C18 preparation. The mobile phase was a 1% acetonitrile - water system containing 0.1% formic acid. The F1 and F3 target segments containing the target substances were collected. The two collected solutions were concentrated to 5% of the original volume at 50 °C, and then refined using an acetonitrile system. The mobile phase was 1% acetonitrile - water. The qualified solution was collected, concentrated to an appropriate volume at 50 °C, and then freeze - dried to obtain 1.67 g of solid F1 and 4.24 g of solid F3. Through structural analysis, F1 was determined to be 1 - galloylglucose, and F3 was phyllemblin F.

[0014] Use of a penthorum chinense compound as described above in the preparation of foods, health products, or drugs for preventing and / or treating ethanol - induced hepatocyte damage.

[0015] Beneficial effects: Compared with the prior art, the advantages of the present invention are as follows:

[0016] Using Gulin penthorum chinense as the raw material, extracting by methanol reflux method, and then obtaining 4 active substances through multi - stage separation and directional collection and purification by preparative liquid chromatography. Through nuclear magnetic resonance and mass spectrometry data analysis, they were finally identified as 1 - galloylglucose, phyllemblin F, xylocalyxin, and a new penthorum chinense compound. The new penthorum chinense compound is tentatively named penthorum chinense glycoside E.

[0017] Through experiments, it was found that the new penthorum chinense compound, penthorum chinense glycoside E, can have a good therapeutic effect on ethanol - induced hepatocyte diseases and potentially treat alcoholic liver diseases. For example, alcoholic liver diseases (such as fatty liver, hepatitis, cirrhosis) are common liver diseases globally. If the safety and effectiveness of penthorum chinense flavone are verified, it can become a new option for clinical treatment, providing a more effective intervention means for alcoholics or patients with alcoholic liver diseases, and reducing the incidence of serious complications such as liver fibrosis and liver failure.

[0018] Reducing liver injury - related complications, ethanol - induced hepatocyte injury may trigger systemic inflammatory responses, abnormal liver function, etc., and then affect other organs (such as the kidneys, nervous system). Protecting hepatocytes can indirectly reduce the risk of multiple organ dysfunction and improve the overall prognosis of patients.

[0019] The possibility of combination therapy, in which the Senecio scandens Buch.-Ham. ex D. Don compounds can be used in combination with existing hepatoprotective drugs (such as polyene phosphatidylcholine, silymarin, etc.) to enhance the therapeutic effect, or reduce the dosage and side effects of existing drugs, providing more options for individualized treatment regimens. Brief Description of the Drawings

[0020] Figure 1 It is a schematic diagram of the molecular structure of the Senecio scandens Buch.-Ham. ex D. Don compounds proposed by the present invention;

[0021] Figure 2 It is a schematic diagram of the HMBC data of the Senecio scandens Buch.-Ham. ex D. Don compounds proposed by the present invention;

[0022] Figure 3 It is a schematic diagram of the 1H-1H COSY data of the Senecio scandens Buch.-Ham. ex D. Don compounds proposed by the present invention;

[0023] Figure 4 It is a schematic diagram of the molecular structure of 1-O-galloylglucopyranose of the Senecio scandens Buch.-Ham. ex D. Don compounds proposed by the present invention;

[0024] Figure 5 It is a schematic diagram of the molecular structure of phyllemblin F of the Senecio scandens Buch.-Ham. ex D. Don compounds proposed by the present invention;

[0025] Figure 6 It is a schematic diagram of the molecular structure of casuarine of the Senecio scandens Buch.-Ham. ex D. Don compounds proposed by the present invention. Detailed Description of the Embodiments

[0026] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention; obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0027] Example 1

[0028] Referring to Figure 1 , this application example provides a Senecio scandens Buch.-Ham. ex D. Don compound, including Figure 1 the structure shown.

[0029] Example 2

[0030] A preparation method of a Senecio scandens Buch.-Ham. ex D. Don compound, using Gulin Senecio scandens Buch.-Ham. ex D. Don as the raw material, extracting by methanol reflux method, and then obtaining 4 active substances through multi-stage separation and directional collection and purification by preparative liquid chromatography. Through nuclear magnetic resonance and mass spectrometry data analysis, they are finally identified as 1-galloylglucose, phyllemblin F, casuarine, and 1 new Senecio scandens Buch.-Ham. ex D. Don compound, and the new Senecio scandens Buch.-Ham. ex D. Don compound is tentatively named as Senecio scandens glycoside E in Chinese.

[0031] 10 kg of Penthorum chinense Pursh herbs were crushed, extracted 3 times with 10 times the amount of 70% methanol-water at 50 °C for 1.5 hours each time, filtered, the filtrate was concentrated to 10 L at 50 °C, then diluted with 1 time the amount of water and filtered. Coarse separation was carried out by C18 preparation. Using a preparative liquid chromatography system filled with C18 packing material, the raw material was roughly separated. The preparation process included column equilibration, sample injection, mobile phase elution, collection of eluent, detection wavelength 280 nm, and the mobile phase was a 5% acetonitrile-water system containing 0.1% formic acid, which was divided into 3 segments, namely acetonitrile crude segment 1, acetonitrile crude segment 2, and acetonitrile crude segment 3. According to the typical chromatogram of gallic acid-glucoside compounds, the elution order was sorted by time and recorded as acetonitrile crude segment one, acetonitrile crude segment two, and acetonitrile crude segment three. The collected solution was detected and judged by an analytical liquid chromatography system, and the qualified solution that met the quality requirements was selected.

[0032] Acetonitrile crude segment 1: Concentrated to 2 L at 50 °C, filtered, and separated again by C18 preparation. The mobile phase was a 1% acetonitrile-water system containing 0.1% formic acid. The target segments F1 and F3 containing the target substance were collected. The two collected solutions were concentrated to 5% of the original volume at 50 °C, and then refined by an acetonitrile system. The mobile phase was 1% acetonitrile-water, and the qualified solution was collected. After concentrating to an appropriate volume at 50 °C, it was freeze-dried to obtain 1.67 g of solid F1 and 4.24 g of solid F3. After structural analysis, F1 was determined to be 1-galloylglucose, and F3 was determined to be phyllemblin F.

[0033] Acetonitrile crude segment 2: Concentrated to 1 L at 50 °C, filtered, and prepared again by C18. The mobile phase was a 5% acetonitrile-water system containing 0.1% formic acid for preparation, detection wavelength 280 nm, and the target segment F4 containing the target was collected. It was concentrated to a small volume at 50 °C, and then repeatedly prepared and separated by a 14% methanol-water system and a 5% acetonitrile-water system. The qualified F4 and F4-2 solutions were collected. After concentration under reduced pressure at 50 °C and then freeze-dried, 9.8 g of solid F4 and 0.369 g of solid F4-2 were obtained. After structural analysis, F4 was determined to be casuarinin, and F4-2 was a new Penthorum chinense Pursh compound, named penthorin E.

[0034] Structural identification results of the active substances in Penthorum chinense Pursh:

[0035] F1: 1-O-Galloyl-β-D-glucopyranose.

[0036] White powder, the quasi-molecular ion peak of LC-MS is m / z = 331.34 [M-H]-, and the molecular formula is C13H16O10.

[0037] 1H-NMR (400 MHz, Methonal-d4): δ 7.12 (2H, s, H-2', H-6'), 5.65 (1H, d, 7.6, H-1), 3.85 (1H, dd, 1.5, 12.1, H-6), 3.70 (1H, dd, J = 4.5, 12.1 Hz, H-6), 3.40~3.50 (4H, m, H-2, H-3, H-4, H-5). 13C-NMR (100 MHz, Methonal-d4) δ: 95.9 (C-1), 74.1 (C-2), 78.2 (C-3), 71.1 (C-4), 78.8 (C-5), 62.3 (C-6), 120.7 (C-1'), 110.5 (C-2'), 146.5 (C-3'), 140.3 (C-4'), 146.5 (C-5'), 110.5 (C-6'), 167.0 (C-7). The above 1H NMR and 13C NMR data are basically consistent with those reported in the literature for 1-O-galloylglucopyranose, so it was identified as 1-O-galloylglucopyranose. Its molecular formula is as Figure 4 shown.

[0038] F3: Phyllanemblinin F.

[0039] White powder, with the quasi-molecular ion peak m / z = 669.40 [M-H]- by LC-MS, and the molecular formula is C27H26O20.

[0040] 1H NMR (400 MHz, Methanol-d4) δ: 7.18 (2H, s, galloyl H-2’,6’), 7.05 (1H, s, neochebuloyl H-3’’), 5.65 (1H, d, 8.0, glc H-1), 5.29 (1H, d, 1.3, neochebuloyl H-2’’’), 4.50 (1H, dd, 2.3, 12.1, H-6a), 4.35 (1H, dd, 5.2, 12.0, H-6b), 3.91 (1H, dd, 1.3, 8.8, H-3’’’), 3.20 (1H, m, H-4’’’), 2.91 (1H, m, H-5’’’a), 2.42 (1H, dd, 4.9, 17.2, H-5’’’b), 13C NMR (100 MHz, Methanol-d4) δ: 175.2, 174.6, 172.3, 166.8, 164.4, (neochebuloyl C-1’’’,6’’’,7’’’,7’’, galloyl C-7’), 146.8 (C-6’’), 146.4 (C-3’,5’), 144.6 (C-4’’), 140.6 (C-5’’), 140.3 (C-4’), 120.7 (C-1’), 117.7 (C-2’’), 116.2 (C-1’’), 110.7 (C-2’,6’), 109.3 (C-3’’), 95.9 (C-1), 78.6 (C-2’’’), 78.0 (C-5), 76.0 (C-3), 74.0 (C-2), 71.1 (C-4), 64.9 (C-6), 45.6 (C-3’’’), 37.2 (C-4’’’), 35.0 (C-5’’’). The above 1H NMR and 13C NMR data are basically consistent with those reported in the literature. Therefore, this compound was identified as Phyllanemblinin F. Its molecular formula is as Figure 5 shown. F4: Strictinin, also known as Phyllanthostrictinin, Minustrictinin, and Puerlipidone (Strictinin).

[0041] White powder, with the quasi-molecular ion peak m / z = 634.01 [M-H]- by LC-MS, and the molecular formula C27H22O18.

[0042] 1H NMR (400 MHz, Methanol-d4) δ: 7.15 (2H, s, H-2″,6″), 6.70- (1H, s,H-2'), 6.56 (1H, s, H-6'), 5.68 (1H, d, J = 8.1 Hz, H-1), 5.24 (1H, dd, J =13.2, 6.3 Hz, H-6), 4.87 (1H, overlap), 4.05 (1H,dd, 9.9, 5.9, H-5), 3.82(1H, d, J = 13.0 Hz, H-3), 3.73 (1H, t, 9.3, H-6), 3.63 (1H, m, H-2); 13C NMR(100 MHz, Methanol-d4) δ 169.9 (C-7'), 169.6 (C-7'''), 166.8 (C-7''), 146.5(C-3', 5'), 145.8 (C-6'', 6'''), 144.8(C-4''), 144.8(C- 4'''), 140.4 (C-4'),137.6 (C-5''),137.3 (C-5'''), 126.5 (C-1''), 126.3 (C- 1'''), 120.5 (C-1'),116.8 (C-2''), 116.6 (C- 2'''), 110.5(C-2', 6'), 108.6 (C-3''), 108.6 (C-3'''), 96.2 (C-1), 76.0 (C-2), 74.7 (C-3), 73.2 (C-4), 73.6 (C-5), 64.2 (C-6). The above 1H NMR and 13C NMR data are basically consistent with those reported in the literature. Therefore, the compound was identified as penthorin. Its molecular formula is as Figure 6 shown.

[0043] F4-2: Penchinensin A.

[0044] This penthorum compound was isolated from the leaves of Penthorum chinense Pursh (Penthorum leaves), a plant of the genus Penthorum in the Saxifragaceae family. After a preliminary literature search, it is a new penthorum compound. For the convenience of description, it is tentatively named penthorin E in Chinese and Penchinensin A in English. As Figure 1 shown.

[0045] White powder, with a quasi-molecular ion peak of m / z = 669.40 [M-H]- by LC-MS, and a molecular formula of C27H26O20.

[0046] 1H NMR (Methanol-d4, 400 MHz) shows signals of three benzene ring hydrogen protons, δ 7.14 (2H, s, H-2''',6''') and 7.02 (1H, s, H-7), one anomeric hydrogen proton signal of β-configured glucose at δ 5.75 (1H, d, 8.2, H-1') and other hydrogen proton signals of the glucosyl group at δ 5.06 (1H, t, 9.6, H-4'), δ 3.83 (1H,t, 9.3, H-3'), δ 3.55 - 3.78 (4H, m, H-2', H-5' and H-6'), one oxygen-linked methine H signal at δ 5.27 (1H, d, 8.3, H-3), two methine H signals at δ 4.81 (1H,overlap, H-4) and 3.72 (1H, m,H-11) and one methylene H signal at δ 2.73 (1H, dd, 17.2, 5.0, H-12a), 2.66 (1H, dd, 17.1,6.4, H-12b).

[0047] 13C NMR (Methanol-d4, 100 MHz) showed 27 carbon signals. There were 5 carbonyl carbon signals at δ 179.8, 174.7, 173.8, 168.7, 166.9; 12 benzene ring carbon signals at δ 146.5×2, 146.4, 145.6, 140.4, 138.9, 122.9, 120.6, 118.4, 111.1, 110.5×2; 7 oxygenated methylene or methine carbon signals at δ 95.8, 79.4, 76.6, 75.8, 74.3, 72.4, 62.0 and 3 methylene or methine carbon signals at δ 44.3, 42.6, 35.0. Combining the above data, it was determined that the compound contained 1 β-glucosyl fragment [δH 5.75 (1H, d, 8.2, H-1’), 3.83 (1H, t, 9.3, H-3’), 3.55 - 3.78 (4H, m, H-2’, H-5’ and H-6’) and δC 95.8, 76.6, 75.8, 74.3, 72.4, 62.0], 1 1,3,4,5-tetrasubstituted galloyl fragment [δH 7.14 (2H, s, H-2’’’,6’’’) and δC 166.9, 146.5×2, 140.4, 120.6, 110.5×2], 1 1,2,3,4,5-pentasubstituted galloyl fragment [δH 7.02 (1H, s, H-7) and δC 168.7, 146.4,145.6, 138.9, 122.9, 118.4, 111.1]. According to the HNMR and combined with the HSQC spectrum, it was determined that C-3 [δH 5.27 and δC 79.4], C-4 [δH 4.81 and δC 44.3] and C-11 [δH 3.72 and δC 42.6] were methine carbon signals, and C-12 [δH 2.73, 2.66 and δC 35.0] was a methylene carbon signal.

[0048] In the 1H-1H COSY spectrum, it was obvious that there were cross-correlations between H-12 / H-11, H-11 / H-4, H-4 / H-3. Combining with H-12 / C-13, H-12 / C-14, H-12 / C-4, H-11 / H-13, H-11 / H-12, H-4 / 12, H-4 / C-12, H-4 / C-2, H-4 / C-11, H-3 / C-4 in the HMBC, the connection order of C-3, C-4, C-11 and C-12 could be determined.

[0049] The correlation between H-1’ / C-7’’ in the HMBC spectrum can determine that the 1,3,4,5-tetrasubstituted galloyl group is acyl-linked to the C-1’ position of the glucosyl group; the correlation between H-4’ / C-15 can determine that the 1,2,3,4,5-pentasubstituted galloyl group is acyl-linked to the C-4’ position of the glucosyl group; the correlations of H-3 / C-5, H-4 / C-5, and H-11 / C-5 can determine that C-4 and C-5 are directly connected. The configurations of C-3, C-4, and C-11 of this Senecio scandens Buch.-Ham. compound cannot be determined. This Senecio scandens Buch.-Ham. compound is a new compound, named Senecio scandens Buch.-Ham. glycoside E. The main HMBC and 1H-1H COSY data are as Figure 2 and Figure 3 shown.

[0050] Example 3

[0051] Use of a Senecio scandens Buch.-Ham. compound as described above in the preparation of a food, health product, or drug for preventing and / or treating ethanol-induced hepatocyte damage.

[0052] The above Senecio scandens Buch.-Ham. compound was used to conduct experiments on ethanol-induced hepatocytes, and the experimental data are as follows:

[0053] Statistical analysis was performed using IBM SPSS Statistics 22.0 software, and one-way ANOVA was used for comparative analysis:

[0054] Table 1 Changes in ALT in cells of each group ( ±SD)

[0055] Group Number of samples (example) ALT (U / L) Blank group 3 10.411±1.358 Model group 3 <![CDATA[20.760±1.840 *** > Model + F1 group 3 <![CDATA[15.365±0.343 ### > Model + F3 group 3 <![CDATA[14.615±1.886 ### > Model + F4 group 3 18.912±1.411 Model + F4-2 group 3 <![CDATA[15.515±2.282 ### >

[0056] Note: Compared with the blank group, in the model group, * P < 0.05, ** P < 0.01, *** P < 0.001;

[0057] Compared with the model group, in the remaining groups, # P < 0.05, ## P < 0.01, ### P < 0.001;

[0058] The statistical results in Table 1 show that compared with the blank group, the alanine aminotransferase (ALT) in the cells of the model group increased (P < 0.001). Among them, the model group was the ethanol-induced hepatocyte damage group. Human hepatocellular carcinoma cells: code HepG2, catalog number icell-h092, icell.

[0059] Compared with the model group, the alanine ammonia ALT in the cells of the model + F1 group, model + F3 group, model + F4 group, and model + F4-2 group decreased (P < 0.001).

[0060] Table 2 Changes of AST in cells of each group ( ±SD)

[0061] Group Number of samples (example) AST (U / L) Blank group 3 35.009±4.620 Model group 3 <![CDATA[60.594±4.143 *** > Model + F1 group 3 <![CDATA[48.754±3.995 # > Model + F3 group 3 <![CDATA[41.670±7.452 ## > Model + F4 group 3 <![CDATA[46.331±2.593 ## <!-- 5 -->]]> Model + F4-2 group 3 <![CDATA[45.170±4.624 ## >

[0062] Note: Compared with the blank group, the model group * P<0.05, ** P<0.01, *** P < 0.001;

[0063] Compared with the model group, the other groups # P<0.05, ## P<0.01, ### P < 0.001;

[0064] The statistical results in Table 2 showed that compared with the blank group, the aspartate aminotransferase (AST) in the cells of the model group was increased (P<0.001).

[0065] Compared with the model group, the AST in the cells of the model+F1 group was decreased (P<0.05), and the AST in the cells of the model+F3 group, model+F4 group, and model+F4-2 group was decreased (P<0.01).

[0066] Table 3 Changes of LDH in cells of each group ( ±SD)

[0067] Group Number of samples (example) LDH (U / gprot) Blank group 3 3527.104±28.847 Model group 3 <![CDATA[5434.782±382.677 *** > Model + F1 group 3 <![CDATA[4421.666±347.759 ### > Model + F3 group 3 <![CDATA[4397.538±335.541 ### > Model + F4 group 3 5155.494±267.119 Model + F4-2 group 3 <![CDATA[4406.431±179.905 ### >

[0068] Note: Compared with the blank group, the model group * P<0.05, ** P<0.01, *** P < 0.001;

[0069] Compared with the model group, the other groups # P<0.05, ## P<0.01, ### P < 0.001;

[0070] The statistical results in Table 3 showed that compared with the blank group, the lactate dehydrogenase (LDH) in the cells of the model group was increased (P<0.001).

[0071] Compared with the model group, the LDH levels in the model+F1, model+F3, and F4-2 groups were decreased (P<0.001), while there was no significant difference in the LDH levels in the model+F4 group (P>0.05).

[0072] Statistics were performed using IBM SPSS Statistics 22.0 software, and comparative analysis was performed using one-way ANOVA.

[0073] Table 4 Changes in SOD in cells of each group ( ±SD)

[0074] Group Number of samples (example) SOD (U / mgprot) Blank group 3 39.452±1.060 Model group 3 <![CDATA[28.794±2.712 *** > Model + F1 group 3 <![CDATA[33.243±1.024 # > Model + F3 group 3 <![CDATA[36.706±3.151 ### > Model + F4 group 3 <![CDATA[32.733±1.323 # > Model + F4-2 group 3 <![CDATA[34.817±2.030 ## >

[0075] Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001;

[0076] Compared with the model group, # P < 0.05, ## P < 0.01, ### P < 0.001;

[0077] The statistical results in Table 4 show that compared with the blank group, SOD in the cells of the model group decreased (P < 0.001).

[0078] Compared with the model group, SOD increased in the cells of the model + F1 group, model + F4 group, and model + F4-2 group (P < 0.05), and SOD increased in the cells of the model + F3 group (P < 0.001).

[0079] Table 5 Changes in MDA in cells of each group ( ±SD)

[0080] Group Number of samples (example) MDA (nmol / mgprot) Blank group 3 1.837±0.307 Model group 3 <![CDATA[3.406±0.225 *** > Model + F1 group 3 <![CDATA[2.095±0.414 ## > Model + F3 group 3 <![CDATA[2.459±0.605 # > Model + F4 group 3 2.966±0.444 Model + F4-2 group 3 <![CDATA[2.459±0.508 # >

[0081] Note: Compared with the blank group, * P < 0.05, ** P < 0.01, *** P < 0.001;

[0082] Compared with the model group, # P < 0.05, ## P < 0.01, ### P < 0.001;

[0083] The statistical results in Table 5 show that compared with the blank group, malondialdehyde (MDA) in the cells of the model group increased (P < 0.001).

[0084] Compared with the model group, MDA decreased significantly in the cells of the model + F1 group (P < 0.01), MDA decreased in the cells of the model + F3 group and model + F4-2 group (P < 0.05), and there was no significant difference in MDA in the cells of the model + F4 group (P > 0.05).

[0085] Table 6 Changes in GSH in cells of each group ( ±SD)

[0086] Group Number of samples (example) GSH (μmol / gprot) Blank group 3 126.433±6.870 Model group 3 <![CDATA[87.305±6.979 *** > Model + F1 group 3 <![CDATA[99.661±5.216 # > Model + F3 group 3 <![CDATA[103.301±5.487 ## > Model + F4 group 3 <![CDATA[100.669±4.852 # > Model + F4-2 group 3 <![CDATA[105.993±7.296 ## >

[0087] Note: Compared with the blank group, the * P < 0.05, ** P < 0.01, *** P < 0.001;

[0088] Compared with the model group, the other # P < 0.05, ## P < 0.01, ### P < 0.001;

[0089] Table 6 Statistical results show that compared with the blank group, glutathione (GSH) in the cells of the model group decreased (P < 0.001).

[0090] Compared with the model group, GSH increased in the cells of the model + F1 group and the model + F4 group (P < 0.05), and GSH increased in the cells of the model + F3 group and the model + F4-2 group (P < 0.01).

[0091] Table 7 Changes in CAT in the cells of each group ( ±SD)

[0092] Group Number of samples (example) CAT (U / mgprot) Blank group 3 6.511±0.484 Model group 3 <![CDATA[3.833±0.208 *** > Model + F1 group 3 <![CDATA[4.920±0.516 # > Model + F3 group 3 <![CDATA[5.294±0.551 ## > Model + F4 group 3 4.405±0.331 Model + F4-2 group 3 <![CDATA[4.959±0.329 # >

[0093] Note: Compared with the blank group, the * P < 0.05, ** P < 0.01, *** P < 0.001;

[0094] Compared with the model group, the other # P < 0.05, ## P < 0.01, ### P < 0.001;

[0095] Table 7 Statistical results show that compared with the blank group, catalase (CAT) in the cells of the model group decreased (P < 0.001).

[0096] Compared with the model group, CAT increased in the cells of the model + F1 group and the model + F4-2 group (P < 0.05), CAT increased in the cells of the model + F3 group (P < 0.01), and there was no significant difference in CAT in the cells of the model + F4 group (P > 0.05).

[0097] In practical applications, it has the following advantages.

[0098] I. Significance for the study of liver diseases

[0099] Revealing the mechanism of liver injury: The protective effect of this Phyllanthus confertus compound may help scientists further clarify the specific molecular mechanisms of ethanol-induced hepatocyte injury (such as oxidative stress, inflammatory response, apoptosis pathway, etc.). By studying the interaction between the Phyllanthus confertus compound and its targets, the pathological process of liver injury can be deeply understood, providing a new perspective for subsequent research.

[0100] Expanding the research direction of liver protection drugs: Currently, there are limited specific drugs for alcoholic liver injury. This Phyllanthus confertus compound may become a lead compound for new liver protection drugs. Its target or pathway (such as activation of antioxidant enzymes, inhibition of inflammatory factors, etc.) can be used as a key basis for drug design, promoting the research and development process of related drugs.

[0101] II. Significance for clinical treatment

[0102] Potential drugs for the treatment of alcoholic liver disease: Alcoholic liver disease (such as fatty liver, hepatitis, cirrhosis) is a common liver disease globally. If this Phyllanthus confertus compound can pass the safety and efficacy verification, it can become a new choice for clinical treatment, providing more effective intervention measures for alcoholics or patients with alcoholic liver disease, and reducing the incidence of serious complications such as liver fibrosis and liver failure.

[0103] Reducing liver injury-related complications: Ethanol-induced hepatocyte injury may trigger systemic inflammatory responses, abnormal liver function, etc., which in turn affect other organs (such as the kidneys, nervous system). Protecting hepatocytes can indirectly reduce the risk of multiple organ dysfunction and improve the overall prognosis of patients.

[0104] Possibility of combination therapy: This Phyllanthus confertus compound can be used in combination with existing liver protection drugs (such as polyene phosphatidylcholine, silymarin, etc.) to enhance the therapeutic effect, or reduce the dosage and side effects of existing drugs, providing more options for individualized treatment regimens.

[0105] III. Significance for public health

[0106] Reducing the social burden of alcohol-related liver diseases: Liver injury caused by alcohol consumption is one of the major global health problems. According to World Health Organization data, alcoholic liver disease accounts for 30% - 50% of the causes of cirrhosis. If this Phyllanthus confertus compound can be translated into practical applications, it can effectively reduce the incidence and mortality of alcoholic liver disease, reduce the consumption of medical resources, and alleviate the burden on families and society.

[0107] Potential for preventive application: For people who need to be exposed to alcohol for a long time (such as alcoholics, alcohol industry practitioners), this Phyllanthus confertus compound may be used as a preventive drug to reduce the risk of liver injury, achieve the goal of "preventing disease before it occurs", and improve the public health level.

[0108] IV. Significance for drug development and the industry

[0109] Promote the industrialization of innovative drugs: If the swertia compound has good drug properties (such as high stability, high bioavailability, and low side effects), it can attract pharmaceutical companies to invest in research and development, form new pharmaceutical products, promote innovation in the pharmaceutical industry, and create economic value.

[0110] Expand the application of natural products or synthetic swertia compounds: If the swertia compound is derived from natural products (such as plants, microorganisms), it can promote the development and utilization of natural drugs; if it is a synthetic swertia compound, the synthesis route can be optimized to reduce production costs and lay a foundation for large-scale production.

[0111] V. Significance for Basic Sciences and Interdisciplinary Research

[0112] Enrich the theory of cell protection mechanisms: This research may discover new cell protection pathways or targets, provide new theories for fields such as cell biology and pharmacology, and promote the development of basic sciences. For example, if the swertia compound exerts its effect by regulating the gut microbiota-liver axis, it can expand the research on the "gut-liver axis" in liver injury.

[0113] Promote interdisciplinary cooperation: From the screening of swertia compounds, mechanism research to clinical translation, multidisciplinary collaboration is required (such as medicinal chemistry, molecular biology, clinical medicine, epidemiology, etc.). This research can promote the cross-integration of different fields and accelerate the transformation of scientific research results.

[0114] VI. Limitations and Future Directions

[0115] Although this discovery is of great significance, further verification is still needed: Safety assessment: Animal toxicology experiments and clinical trials are required to confirm the safety and tolerance of swertia compounds in vivo.

[0116] In-depth study of the mechanism of action: Identify key targets and signaling pathways to avoid the problem of unclear mechanisms caused by "pleiotropy".

[0117] Explore clinical applicability: Stratified studies are required for the efficacy differences in different populations (such as different alcohol consumption levels and stages of liver injury).

[0118] The above are only specific implementation manners of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A kind of Penthorum chinense Pursh compound, characterized in that, It has the structure described in Formula 1: Formula 1.

2. A preparation method of the swertia compound as described in claim 1, characterized in that, It includes: The herba penthori is crushed, and a 70% methanol-water mixture 10 times the amount of herba penthori is added, heated and extracted 3 times at 50 °C, each heating for 1.5 hours, filtered, the filtrate is concentrated to 10 L, and then diluted with one-fold water and filtered; Coarse separation by C18 preparation. Using a preparative liquid phase system filled with C18 packing, the raw materials are coarsely separated. The preparation process includes column equilibration, sample injection, mobile phase elution, collection of eluent, detection wavelength is 280 nm, and the mobile phase is an acetonitrile-water system with 0.1% formic acid, including three segments: acetonitrile coarse segment 1, acetonitrile coarse segment 2, and acetonitrile coarse segment 3. According to the typical chromatogram of gallic acid-glucoside compounds, the elution peak order is sorted by time and recorded as acetonitrile coarse segment 1, acetonitrile coarse segment, and acetonitrile coarse segment 3. The collected solution is detected and judged by an analytical liquid phase system, and the solution meeting the quality requirements is screened out and judged as a qualified solution; The acetonitrile coarse segment 2 is concentrated to 1 L at 50 °C, filtered, and C18 is prepared again. The mobile phase is prepared with a 5% acetonitrile-water system containing 0.1% formic acid, the detection wavelength is 280 nm, the target F4 segment is collected, concentrated to a small volume at 50 °C, and repeatedly prepared and separated with a 14% methanol-water system and a 5% acetonitrile-water system. The qualified F4 and F4-2 solutions are collected, concentrated under reduced pressure at 50 °C and then freeze-dried to obtain F4 solid and F4-2 solid, wherein F4 is casuarine, and F4-2 is the penthorum chinense pursh compound described in claim 1.

3. Use of the penthorum chinense pursh compound described in claim 1 in the preparation of a medicament for preventing and / or treating ethanol-induced hepatocyte injury.

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