Polyphenols and polyhydroxy furan compounds in black mulberry fruits, extraction method and application thereof

Abstract

This invention discloses polyphenols and polyhydroxyfurans from black mulberry fruit, their extraction methods, and applications, belonging to the field of traditional Chinese medicine extraction. Specifically, it relates to seven polyphenols and one polyhydroxyfuran compound isolated from black mulberry fruit, and their extraction methods. It also provides the effects of these compounds or their pharmaceutically acceptable salts, or pharmaceutical compositions containing these compounds, on the survival rate of mouse glial cells BV-2 after H2O2 modeling, which can be used to prepare antioxidant drugs. This invention further enriches the structural diversity of active substances in black mulberry, laying the foundation for subsequent bioactivity testing of the obtained monomeric compounds, providing active lead compounds for new drug development, and also providing a theoretical basis for in-depth research and development of black mulberry.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine extraction, specifically relating to a polyphenol and polyhydroxy furan compound isolated from black mulberry fruit, its extraction method, and its application in the preparation of antioxidant drugs. Background Technology

[0002] There are approximately 16 species of plants in the genus *Morus* of the family Moraceae, mainly distributed in the North Temperate Zone. Most of these are found in my country, including *Morus alba*, *Morus sorrata*, *Morus macroura*, *Morus notabilis*, *Morus mongolica*, *Morus australis*, *Morus cathayana*, *Morus nigra*, *Morus liboensis*, *Morus wittiorum*, and *Morus trilobata* (Flora of China Editorial Committee. Flora of China [M]. Beijing: Science Press, 1998, 23: 6-23). ​​The *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) records that many parts of *Morus* plants can be used medicinally; for example, the branches, leaves, fruits, and root bark of *Morus* contain special substances and functional components, possessing unique medicinal value. The 2020 edition of the Pharmacopoeia of the People's Republic of China (Part I) records that the fruit spike of the mulberry tree is called "mulberry," and is sweet, sour, and cold in nature. It enters the heart, liver, and kidney meridians. It has the effects of nourishing yin and blood, promoting body fluid and moistening dryness. It is often used to treat symptoms such as liver and kidney yin deficiency, dizziness and tinnitus, palpitations and insomnia, premature graying of hair, thirst due to fluid depletion, internal heat and thirst, and constipation due to intestinal dryness (National Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China. Part I [S]. Beijing: China Medical Science and Technology Press, 2020: 313).Black mulberry (Morus nigra), as a major species of mulberry plant distributed in my country, has been the subject of some modern pharmacological research. Published studies have shown that black mulberry possesses antioxidant, antibacterial, antitumor, antiviral, hypotensive, and hypoglycemic effects. (Ercisli S, Orhan E. Chemical composition of white (Morus alba), red (Morus rubra) and black (Morus nigra) mulberry fruits [J]. Food Chemistry, 2007, 103(4): 1380-1384; Fu Daxu, Chen Lei, Hou Aijun, et al. Study on chemical composition of black mulberry [J]. Chinese Traditional and Herbal Drugs, 2005, 36(9): 1296-1299; Wang Lei, Wang Hongqing, Chen Ruoyun. Study on chemical composition of black mulberry stem bark [J]. Chinese Journal of Traditional Chinese Medicine, 2007, 32(23): 2497-2499; Ma XL, Song F, Zhang H, et al. Quantitative determination and comparison of...) Polysaccharide from Morus nigra Linn by HPCE and HPLC[J]. Current Pharmaceutical Analysis, 2017, 13: 433-437; Arfan M, Khan R, Rybarczyk A, et al. Antioxidant activity of mulberry fruit extracts[J]. International Journal of Molecular Sciences, 2012, 13(12): 2472-2480). Zhu Cuiling et al. extracted mulberry with 75% ethanol and determined its anti-inflammatory activity: the cytotoxicity of different concentrations of samples on RAW264.7 cells, the effect of different concentrations of mulberry extract on LPS-induced expression of iNOS and COX-2 mRNA in RAW264.7 cells, and the detection of iNOS and COX-2 protein expression levels in RAW264.7 cells by Western Blot.Experimental results showed that mulberry extract could inhibit LPS-induced secretion of NO and PGE2 in RAW264.7 cells, inhibit the expression of COX-2 and iNOS mRNA, and inhibit the expression of iNOS and COX-2 proteins. These inhibitory effects showed a certain concentration dependence (Zhu Cuiling, Chen Ming, Wang Menghan, Shen Ting, Qiang Qian, Wang Xinfeng, Ji Lilian, Feng Zuoshan, Tao Yongxia, Bai Yujia, Hu Weicheng. Study on the in vitro anti-inflammatory effects and mechanisms of mulberry extract [J]. Modern Food Science and Technology, 2017, 33(04):61-66+37). In addition, Xiang Wei et al. extracted mulberry branches from Xinjiang using 70% ethanol cold soaking and isolated a class of flavonoid compounds with antioxidant activity, mulberry root ketone A, from the extract. The authors used the antioxidant BHT (2,6-di-tert-butyl-4-methylphenol) as a control to determine the scavenging of DPPH free radicals, OH free radicals, and total reducing power. The results showed that the compound scavenges the IC of DPPH radicals and OH radicals. 50 The values ​​were 50.3 and 96.5 mg / L, respectively (compared to 64.2 and 231.6 mg / L for the control BHT), and the total reducing power was also significantly higher than that of the control (Xiang Wei, Yu Yan, Liu Jing, Xu Li, Huang Xianzheng, Ding Tianlong, Zuo Shaochun. Antioxidant active ingredient sanggenon A in mulberry branches from Xinjiang [C]. Proceedings of the 12th Symposium on Genetics and Breeding of Domestic Silkworms (Abstract Collection), 2016:216). In order to maximize the medicinal value of Curcuma zedoaria, a systematic component study was conducted on the rhizome of Curcuma zedoaria, and new polyphenols and polyhydroxy furans were extracted. The structures of the compounds were confirmed by nuclear magnetic resonance, infrared spectroscopy, and mass spectrometry, and their anti-inflammatory activities were detected. Summary of the Invention

[0003] The primary objective of this invention is to provide seven polyphenols and one polyhydroxyfuran compound.

[0004] A second objective of this invention is to provide a method for extracting seven polyphenols and one polyhydroxyfuran compound.

[0005] A third object of the present invention is to provide a pharmaceutical composition containing seven polyphenols and one polyhydroxyfuran compound.

[0006] A fourth object of the present invention is to provide the use of seven polyphenols and one polyhydroxy furan compound or isomers thereof, pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising such compounds in the preparation of antioxidant medicaments.

[0007] The technical solution of this invention is summarized as follows:

[0008] The seven polyphenols and one polyhydroxyfuran compound are any one of the following compounds or isomers of the compound, or any one of pharmaceutically acceptable salts of the compound;

[0009]

[0010] The pharmaceutically acceptable salts include sodium salts, potassium salts, ammonium salts, hydrochloride salts, and sulfate salts.

[0011] The isomers include: optical isomers, cis-trans isomers, racemates, and mixtures thereof.

[0012] The present invention also provides a method for extracting the polyphenols and polyhydroxyfuran compounds 1-8, comprising the following steps:

[0013] (1) Using dried fruit of black mulberry as raw material, add 8 to 15 times the mass of the raw material in a 95% ethanol aqueous solution, reflux extract 2 to 4 times, each extraction for 2 to 4 hours, combine the extracts, recover the solvent under reduced pressure, and concentrate to obtain total extract.

[0014] (2) Disperse the total extract in 5 to 10 times its weight of water and extract with ethyl acetate to obtain an ethyl acetate layer and an aqueous layer;

[0015] (3) The ethyl acetate extract concentrate was separated by silica gel column chromatography, using a gradient elution of dichloromethane-methanol with a volume ratio of 100:0 to 0:1. The fractions E1 (100:0), E2 (50:1), E3 (30:1), E4 (20:1), E5 (10:1), E6 (6:1), E7 (4:1), E8 (2:1), and E9 (0:1) were collected.

[0016] (4) The concentrated fraction E7 was separated by Sephadex LH-20 column chromatography (CH3OH) and further purified to obtain compounds 1-4 and 6-8;

[0017] (5) The concentrated fraction E8 was separated by silica gel column chromatography with dichloromethane-methanol and further purified to obtain compounds 5 and 7.

[0018] The specific separation and purification process for fractions E7 and E8 is as follows:

[0019] Fraction E7 was concentrated and then separated by Sephadex LH-20 column chromatography (CH3OH) to obtain fraction E72;

[0020] After concentration, fraction E72 was separated by ODS column chromatography with a gradient elution of methanol-water with a volume ratio of 2:8 to 10:0. Fractions with volume ratios of 2:8 and 6:4 were collected and designated as E721 and E723.

[0021] After concentration, fraction E721 was separated by silica gel column chromatography and eluted with dichloromethane-methanol at a volume ratio of 20:1 to 1:1. Fractions with a volume ratio of dichloromethane to methanol of 15:1 and 5:1 were collected and designated as E7213 and E7215, respectively.

[0022] Fraction E7213 was concentrated and purified by preparative HPLC with methanol-water (30:70 v / v) as the mobile phase to obtain compounds 2, 3, 6 and 7.

[0023] Fraction E7215 was purified by preparative HPLC with methanol-water (v / v) as the mobile phase (v / v) to obtain compound 4.

[0024] After concentration, fraction E723 was separated by silica gel column chromatography and eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1. The fraction with a volume ratio of dichloromethane to methanol of 10:1 was collected and denoted as E7235.

[0025] Fraction E7235 was purified by preparative HPLC using a methanol-water mixture with a volume ratio of 60:40 as the mobile phase to obtain compound 1.

[0026] After concentration, fraction E8 was separated by silica gel column chromatography and eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1. The fraction with a volume ratio of dichloromethane to methanol of 10:1 was collected and denoted as E82.

[0027] After concentration, fraction E82 was separated by ODS column chromatography with a gradient elution of methanol-water with a volume ratio of 2:8 to 10:0. The fraction with a volume ratio of 2:8 was collected and designated as E821.

[0028] After concentration, fraction E821 was separated by silica gel column chromatography and eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1. The fraction with a volume ratio of dichloromethane to methanol of 10:1 was collected and designated as E8213.

[0029] Fraction E8213 was purified by preparative HPLC with methanol-water (30:70 v / v) as the mobile phase to obtain compounds 5 and 8.

[0030] An extract from black mulberry fruit containing polyphenols and polyhydroxyfurans.

[0031] Application of black mulberry fruit extract in the preparation of antioxidant drugs.

[0032] A pharmaceutical composition comprising one or more of the aforementioned polyphenols and polyhydroxyfurans, isomers of these compounds, and pharmaceutically acceptable salts of these compounds; further comprising one or a combination of a pharmaceutically acceptable carrier, excipient, and diluent. The pharmaceutical composition is administered orally or by injection, and in dosage forms including tablets, capsules, powders, syrups, and injections.

[0033] The present invention also provides the use of the aforementioned polyphenols and polyhydroxy furans, isomers of these compounds, pharmaceutically acceptable salts of these compounds, or the pharmaceutical compositions thereof in the preparation of antioxidant drugs.

[0034] Advantages of this invention:

[0035] The polyphenols and polyhydroxyfurans or their isomers, pharmaceutically acceptable salts, or pharmaceutical compositions containing these compounds of the present invention significantly enhance the survival rate of mouse glial cells BV-2 after hydrogen peroxide modeling, and can be used to prepare antioxidant drugs. This invention further enriches the structural diversity of active substances in black mulberry fruit, laying the foundation for subsequent bioactivity testing of the obtained monomeric compounds, providing active lead compounds for new drug development, and also providing a theoretical basis for in-depth research and development of black mulberry fruit. Attached Figure Description

[0036] Figure 1 Effects of compounds 1-8 on the survival rate of mouse glial cells BV-2 after hydrogen peroxide modeling. Detailed Implementation

[0037] The technical solution of the present invention will be further described below with reference to specific embodiments.

[0038] Example 1

[0039] The method for extracting polyphenols and polyhydroxyfurans from black mulberry fruit includes the following steps:

[0040] (1) Using dried fruit of black mulberry as raw material, add 10 times the mass of the raw material with a volume concentration of 95% ethanol aqueous solution, reflux extract 3 times, each extraction for 2 hours, combine the extracts, recover the solvent under reduced pressure, concentrate to obtain total extract (7.7 kg).

[0041] (2) The total extract was dispersed in 8 times its weight of water and extracted with ethyl acetate to obtain an ethyl acetate layer (954 g) and an aqueous layer.

[0042] (3) The ethyl acetate extract concentrate was separated by silica gel column chromatography, using a gradient elution of dichloromethane-methanol with a volume ratio of 100:0 to 0:1. The fractions E1 (100:0), E2 (50:1), E3 (30:1), E4 (20:1), E5 (10:1), E6 (6:1), E7 (4:1), E8 (2:1), and E9 (0:1) were collected.

[0043] Fraction E7 (45.5g) was concentrated and then separated by Sephadex LH-20 column chromatography (CH3OH) to obtain fraction E72;

[0044] After concentration, fraction E72 (21.3g) was separated by ODS column chromatography. The fractions were eluted by a gradient of methanol-water with volume ratios of 2:8, 4:6, 6:4, 7:3, and 10:0. The fractions with volume ratios of 2:8 and 6:4 were collected and designated as E721 and E723.

[0045] Fraction E721 (10.8 g) was concentrated and separated by silica gel column chromatography. It was eluted with dichloromethane-methanol at a volume ratio of 20:1 to 1:1. Fractions with a volume ratio of dichloromethane to methanol of 15:1 and 5:1 were collected and designated as E7213 and E7215, respectively.

[0046] The concentrated fraction E7213 (2.8 g) was purified by preparative HPLC with methanol-water (30:70 v / v) as the mobile phase to give compounds 2 (2.1 mg), 3 (10.4 mg), 6 (2.0 mg) and 7 (2.4 mg).

[0047] Fraction E7215 (810 mg) was purified by preparative HPLC with methanol-water (v / v) as the mobile phase (v / v) to give compound 4 (11.7 mg).

[0048] Fraction E723 (3.8g) was concentrated and separated by silica gel column chromatography. It was eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1. The fraction with a volume ratio of dichloromethane to methanol of 10:1 was collected and denoted as E7235.

[0049] Fraction E7235 was purified by preparative HPLC with methanol-water (60:40 v / v) as the mobile phase to obtain compound 1 (1.8 g).

[0050] Fraction E8 (41.8 g) was concentrated and separated by silica gel column chromatography. It was eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1. The fraction with a volume ratio of dichloromethane to methanol of 10:1 was collected and denoted as E82.

[0051] After concentration, fraction E82 (28.1g) was separated by ODS column chromatography. The fraction was eluted with methanol-water gradient eluent at volume ratios of 2:8, 4:6, 6:4, 7:3, and 10:0. The fraction with a volume ratio of 2:8 was collected and designated as E821.

[0052] The fraction E821 (14.2 g) was concentrated and separated by silica gel column chromatography. It was eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1. The fraction with a volume ratio of dichloromethane to methanol of 10:1 was collected and designated as E8213.

[0053] Fraction E8213 (2.1 g) was concentrated and purified by preparative HPLC with methanol-water (30:70 v / v) as the mobile phase to give compounds 5 (6.4 mg) and 8 (7.4 mg).

[0054] Compound 1 is a yellow amorphous powder. HR-ESI-MS m / z 521.1592 [M+Cl] - (calcd for C 26 H 30 O9 - ,521.1584), molecular formula C 26 H 30 O9, 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 1.

[0055] Compound 2 is a yellow amorphous powder. HR-ESI-MS m / z 280.0829 [MH] - (calcd for C 13 H 14 O6N - ,280.0827), molecular formula C 13 H 15 O6N, 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 2.

[0056] Compound 3 is a yellow amorphous powder. HR-ESI-MS m / z 263.1134 [M+H] + (calcd for C 11 H 19 O7 +,263.1125), molecular formula C 11 H 18 O7, 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 3.

[0057] Compound 4 is a yellow amorphous powder. HR-ESI-MS m / z 267.0850 [M+H] + (calcd for C 13 H 15 O6 + ,267.0863), molecular formula C 13 H 14 O6, 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR(150MHz,DMSO-d6)Table 4.

[0058] Compound 5 is a yellow amorphous powder. HR-ESI-MS m / z 665.2043 [M+Na] + (calcd for C 29 H 38 O 16 Na + ,665.2052), molecular formula C 29 H 38 O 16 , 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 5.

[0059] Compound 6 is a yellow amorphous powder. HR-ESI-MS m / z 513.1753 [M+Na] + (calcd for C 25 H 30 O 10 Na + ,513.1731), molecular formula C 25 H 30 O 10 , 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 6.

[0060] Compound 7 is a yellow amorphous powder. HR-ESI-MS m / z 513.1753 [M+Na] + (calcd for C 25 H 30 O 10 Na + ,513.1731), molecular formula C 25 H 30 O 10 , 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 6.

[0061] Compound 8 is a yellow amorphous powder. HR-ESI-MS m / z 473.1065 [M+Na] + (calcd for C 21 H 22 O 11 Na + ,473.1054), molecular formula C 21 H 22 O 11 , 1 H-NMR (600MHz, DMSO-d6) and 13 C-NMR (150MHz, DMSO-d6) data are shown in Table 7.

[0062] Table 1 1 H-NMR and 13 C-NMR data of compound 1(DMSO-d6,δin ppm,J in Hz)

[0063]

[0064]

[0065] Table 2 1 H-NMR and 13 C-NMR data of compound 2(DMSO-d6,δin ppm,J in Hz)

[0066]

[0067] Table 3 1 H-NMR and 13C-NMR data of compound 3(DMSO-d6,δin ppm,J in Hz)

[0068]

[0069] Table 4 1 H-NMR and 13 C-NMR data of compound 4(DMSO-d6,δin ppm,J in Hz)

[0070]

[0071]

[0072] Table 5 1 H-NMR and 13 C-NMR data of compound 5(CDCl3,δin ppm,J in Hz)

[0073]

[0074] Table 6 1 H-NMR and 13 C-NMR data of compounds 6 and 7(DMSO-d6,δin ppm,Jin Hz)

[0075]

[0076]

[0077] Table 7 1 H-NMR and 13 C-NMR data of compound 7(DMSO-d6,δin ppm,J in Hz)

[0078]

[0079] Using physicochemical data and modern spectroscopic techniques (HRESIMS and NMR), combined with relevant data from published literature, the structures of the above compounds were identified, confirming that compounds 1-8 are all novel compounds not previously reported in the literature, as shown below:

[0080]

[0081]

[0082] Example 2

[0083] (1) Using dried fruit of black mulberry as raw material, add 8 times the mass of the raw material to a 95% ethanol aqueous solution with a volume concentration of 95%, reflux extract twice, extract for 2 hours each time, combine the extracts, recover the solvent under reduced pressure, concentrate to obtain total extract.

[0084] (2) Disperse the total extract in 6 times its mass of water and extract with ethyl acetate to obtain an ethyl acetate layer and an aqueous layer;

[0085] (3) The ethyl acetate extract concentrate was separated by silica gel column chromatography, using a gradient elution of dichloromethane-methanol at a volume ratio of 100:0 to 0:1. Fractions E1 (100:0), E2 (50:1), E3 (30:1), E4 (20:1), E5 (10:1), E6 (6:1), E7 (4:1), E8 (2:1), and E9 (0:1) were collected. Fractions E7 and E8 were further purified to obtain compounds 1-8. The specific separation and purification process was the same as in Example 1.

[0086] The effects of compounds 1-8 extracted above on the survival rate of mouse glial cells BV-2 after H2O2 modeling are studied in detail below:

[0087] 1. Principle: Hydroxyl radicals are highly toxic substances that can participate in various reactions within the body, such as electron transfer and dehydrogenation, causing irreversible oxidative damage to lipids, proteins, and nucleic acids. Hydrogen peroxide, as a strong oxidant, can freely cross cell membranes and react with intracellular F... e 3+ The reaction generates hydroxyl radicals, which induce oxidative stress in the body. H₂O₂ is relatively stable and does not readily decompose in the body, making it widely used to induce oxidative damage models in animal tissues or cells. Oxidative damage in cells is often accompanied by a decrease in cell viability; therefore, detecting cell viability is used to characterize the degree of oxidative damage.

[0088] 2. Methods: Mouse glial cells (BV-2) were cultured in DMEM medium containing 10% fetal bovine serum, 100 μg / mL streptomycin, and 100 U / mL penicillin sodium, and incubated at 37℃ in a 5% CO2 incubator. BV-2 cells in the logarithmic growth phase were collected and their concentration adjusted to 1×10⁻⁶ cells / mL. 4Cells / wells were seeded in 96-well plates, with 80 μL of cell suspension added to each well. The experiment included a control group (BV-2 cells, DMSO), a model group (BV-2 cells, DMSO, 300 μM H2O2), and a test drug group (BV-2 cells, the test compound, 300 μM H2O2). After cells adhered and grew to 70% confluence, the drug-treated groups were given 20 μL of drug-containing medium, while the control and model groups received the same volume of medium. The cells were incubated in a 5% CO2, 37°C incubator for 24 h. The model and drug-treated groups were given 300 μM H2O2 for 0.5 h, followed by the addition of 10 μL of CCK-8 reagent to each well. The 96-well plates were then incubated for 1–4 h, and the absorbance at 450 nm was measured using a microplate reader. The cell viability formula is as follows:

[0089] Cell viability (%) = [(As-Ab) / (Ac-Ab)] × 100%

[0090] As = Absorbance of experimental wells (absorbance of wells containing cells, culture medium, CCK-8, and the analyte compound)

[0091] Ab = Absorbance of blank wells (absorbance of wells containing culture medium and CCK-8)

[0092] Ac = Absorbance of control wells (absorbance of wells containing cells, culture medium, and CCK-8)

[0093] 3. Results: After the addition of 20 μM compounds 1, 5, and 7, the survival rate of BV-2 glial cells in mice induced by H2O2 significantly increased. Figure 1 ).

[0094] The above description of the embodiments is only for the purpose of helping to understand the method and central idea of ​​the present invention. It should be noted that those skilled in the art can make several improvements and modifications to the present invention without departing from the principle of the present invention, and these improvements and modifications also fall under the protection of the claims of the present invention.

Claims

1. Polyphenols and polyhydroxyfurans in black mulberry fruit, characterized in that, It is a compound as shown below or a pharmaceutically acceptable salt thereof; 。 2. A method for extracting polyphenols and polyhydroxyfurans from the black mulberry fruit as described in claim 1, characterized in that, Includes the following steps: (1) Using dried fruit of black mulberry as raw material, add 8 to 15 times the mass of the raw material in a 95% ethanol aqueous solution, reflux extract 2 to 4 times, each extraction for 2 to 4 hours, combine the extracts, recover the solvent under reduced pressure, concentrate and obtain total extract. (2) Disperse the total extract in 5 to 10 times its weight of water and extract with ethyl acetate to obtain an ethyl acetate layer and an aqueous layer; (3) The ethyl acetate extract concentrate was separated by silica gel column chromatography, using dichloromethane-methanol gradient elution with a volume ratio of 100:0~0:

1. The fractions E1 (100:0), E2 (50:1), E3 (30:1), E4 (20:1), E5 (10:1), E6 (6:1), E7 (4:1), E8 (2:1), and E9 (0:1) were collected. (4) The concentrated fraction E8 was separated by silica gel column chromatography with dichloromethane-methanol and further purified to obtain compound 5.

3. The method for extracting polyphenols and polyhydroxyfurans from black mulberry fruit according to claim 2, characterized in that, The specific separation and purification process of fraction E8 in step (4) is as follows: After concentration, fraction E8 was separated by silica gel column chromatography, eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1, and the fraction was collected and designated as E82. After concentration, fraction E82 was separated by ODS column chromatography, using methanol-water gradient elution with a volume ratio of 2:8 to 10:

0. The fraction was collected and designated as E821. After concentration, fraction E821 was separated by silica gel column chromatography, eluted with dichloromethane-methanol at a volume ratio of 15:1 to 1:1, and the fraction was collected and designated as E8213. Fraction E8213 was purified by preparative HPLC with a mobile phase of methanol-water at a volume ratio of 30:70 to obtain compound 5.

4. A pharmaceutical composition, characterized in that, It comprises one of the polyphenols and polyhydroxyfurans of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier; The drug composition is administered orally or by injection, and the dosage forms include tablets, capsules, powders, syrups, and injections.

5. The use of the pharmaceutical composition of claim 4 in the preparation of an antioxidant drug.

6. The use of the polyphenols and polyhydroxyfurans of claim 1, or pharmaceutically acceptable salts thereof, in the preparation of antioxidant drugs.

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