CN117229337A - Lignan compound containing tannin structure separated from phyllanthus emblica and liver protection application thereof - Google Patents
Lignan compound containing tannin structure separated from phyllanthus emblica and liver protection application thereof Download PDFInfo
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- CN117229337A CN117229337A CN202311201378.3A CN202311201378A CN117229337A CN 117229337 A CN117229337 A CN 117229337A CN 202311201378 A CN202311201378 A CN 202311201378A CN 117229337 A CN117229337 A CN 117229337A
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- 235000015489 Emblica officinalis Nutrition 0.000 title claims abstract description 44
- 229930013686 lignan Natural products 0.000 title claims abstract description 25
- 235000009408 lignans Nutrition 0.000 title claims abstract description 25
- 210000004185 liver Anatomy 0.000 title claims abstract description 25
- 235000018553 tannin Nutrition 0.000 title claims abstract description 24
- 229920001864 tannin Polymers 0.000 title claims abstract description 24
- 239000001648 tannin Substances 0.000 title claims abstract description 24
- -1 Lignan compound Chemical class 0.000 title claims abstract description 23
- 240000009120 Phyllanthus emblica Species 0.000 title claims abstract 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 32
- 239000003814 drug Substances 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 8
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- 206010067125 Liver injury Diseases 0.000 claims abstract description 7
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 48
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 20
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 12
- 238000000605 extraction Methods 0.000 claims description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
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- 238000000746 purification Methods 0.000 claims description 4
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- 238000003810 ethyl acetate extraction Methods 0.000 claims description 3
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- 238000010438 heat treatment Methods 0.000 claims description 3
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
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- FDQAOULAVFHKBX-UHFFFAOYSA-N Isosilybin A Natural products C1=C(O)C(OC)=CC(C2C(OC3=CC(=CC=C3O2)C2C(C(=O)C3=C(O)C=C(O)C=C3O2)O)CO)=C1 FDQAOULAVFHKBX-UHFFFAOYSA-N 0.000 description 3
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- VLGROHBNWZUINI-UHFFFAOYSA-N Silybin Natural products COc1cc(ccc1O)C2OC3C=C(C=CC3OC2CO)C4Oc5cc(O)cc(O)c5C(=O)C4O VLGROHBNWZUINI-UHFFFAOYSA-N 0.000 description 3
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 3
- 239000001768 carboxy methyl cellulose Substances 0.000 description 3
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- 238000011160 research Methods 0.000 description 3
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- SEBFKMXJBCUCAI-HKTJVKLFSA-N silibinin Chemical compound C1=C(O)C(OC)=CC([C@@H]2[C@H](OC3=CC=C(C=C3O2)[C@@H]2[C@H](C(=O)C3=C(O)C=C(O)C=C3O2)O)CO)=C1 SEBFKMXJBCUCAI-HKTJVKLFSA-N 0.000 description 3
- 229940043175 silybin Drugs 0.000 description 3
- 235000014899 silybin Nutrition 0.000 description 3
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 3
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 2
- 108010082126 Alanine transaminase Proteins 0.000 description 2
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 2
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 238000012449 Kunming mouse Methods 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 235000019483 Peanut oil Nutrition 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000009609 fructus phyllanthi Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 2
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 2
- 150000005692 lignans Chemical class 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 239000000312 peanut oil Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 1
- 241000221017 Euphorbiaceae Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000158764 Murraya Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241001130943 Phyllanthus <Aves> Species 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
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- 238000003556 assay Methods 0.000 description 1
- 235000019606 astringent taste Nutrition 0.000 description 1
- 235000015241 bacon Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
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- 238000001311 chemical methods and process Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
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- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
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- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
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- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
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- 210000002784 stomach Anatomy 0.000 description 1
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- 150000003648 triterpenes Chemical class 0.000 description 1
Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a lignan compound containing a tannin structure, which is separated from phyllanthus emblica and has a structure shown in a formula I. The inventors determined CCl by using a fully automatic biochemical analyzer and IFCC method 4 AST and ALT activities in serum of mice with liver injury models are obviously reduced, and the compound has liver protection activity. The invention also discloses application of the lignan compound containing the tannin structure in preparing liver protecting medicines.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a novel lignan compound containing a tannin structure, which is separated from phyllanthus emblica, and a preparation method and a liver protection application of the compound.
Background
Phyllanthus emblica (Phyllanthus emblica L.) is the dried mature fruit of Phyllanthus emblica (Phyllanthus Linn.) of Euphorbiaceae, also known as Emula Murraya, fructus Phyllanthi, phyllanthus emblica, etc. Fructus phyllanthi is used as a medicine and food dual-purpose traditional Chinese medicine, and has been used for more than two thousand years in China. The research shows that the phyllanthus emblica contains rich and diverse chemical components including phenolic acids, tannins, flavonoids, lignans, sterols, lignans, triterpenes, alkaloids and the like. Modern pharmacological researches have found that phyllanthus emblica has better effects of protecting liver, resisting oxidation, resisting tumor, resisting inflammation and aging, reducing blood sugar and blood fat, regulating immunity and the like. Since 1977, phyllanthus emblica is collected in each edition of Chinese pharmacopoeia in turn by using Tibetan conventional medicinal materials, has sweet, sour and astringent taste and cool nature, and has the effects of soothing liver, promoting bile flow, clearing heat, cooling blood, promoting digestion, invigorating stomach, promoting fluid production, relieving cough and the like. The Tibetan medicine is often treated by phyllanthus emblica for treating 'hematopathy', 'red barker' and 'bacon' and has a plurality of similarities with clinical manifestations of liver and gall diseases in modern medicine. Therefore, further development of the potential pharmaceutical value of phyllanthus emblica is necessary.
Disclosure of Invention
The inventor selects phyllanthus emblica which is a plant used as both medicine and food as a research object, adopts a biological activity guiding separation method to separate and purify the liver-protecting active part of the phyllanthus emblica, and obtains a novel lignan compound containing a tannin structure. Meanwhile, the inventors determined CCl by using a full-automatic biochemical analyzer and IFCC method 4 The activity of glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) in serum of a mouse with liver injury model is obviously reduced, and the novel compound has liver protection activity. The present invention aims to provide a novel lignan compound containing a tannin structure isolated from Emblica officinalis, and the chemical process thereofA preparation method of the compound and liver protection activity.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
novel lignan compound containing tannin structure and having a structure shown in formula I and being used for medical application and isolated from phyllanthus emblica:
compound i english name: 1-O-heretol-3, 6-ditolloyl-O- β -D-glucoside; chinese name: 1-O-bordeaux-3, 6-digalliyl-O- β -D-glucoside; molecular formula C 40 H 38 O 19 。
The preparation method of the lignan compound containing the tannin structure comprises the following steps:
crushing the dried phyllanthus emblica, soaking the crushed phyllanthus emblica in 90-100% ethanol for 20-24 hours, heating and refluxing for 3-5 times, each time for 4-6 hours, and combining the filtrates to obtain a phyllanthus emblica total extract;
step (2), carrying out vacuum concentration on the total extract of the phyllanthus emblica to obtain total thick phyllanthus emblica paste;
uniformly suspending the total phyllanthus emblica thick paste in distilled water, and sequentially extracting with petroleum ether, ethyl acetate and n-butanol to obtain petroleum ether extraction sites, ethyl acetate extraction sites and n-butanol extraction sites respectively;
step (4), the n-butanol extraction part is put on a macroporous adsorption resin column, and gradient elution is carried out by using 10% ethanol, 25% ethanol, 45% ethanol, 70% ethanol and 100% ethanol in sequence, so as to obtain 5 main fractions: main fraction A, main fraction B, main fraction C, main fraction D, main fraction E;
and (5) separating the main fraction B on a medium pressure chromatographic column to obtain 3 secondary fractions: secondary fraction B-1, secondary fraction B-2, secondary fraction B-3;
step (6), loading the secondary fraction B-2 on a Toyopearl HW-40C column, eluting with methylene dichloride-methanol volume ratio=1:2 to obtain a subfraction B-2-1 and a subfraction B-2-2;
step (7), loading the subfraction B-2-2 on a Sephadex LH-20 column, eluting with 70-90% methanol to obtain a fine fraction B-2-2-1 and a fine fraction B-2-2-2;
and (8) carrying out preparation liquid chromatography separation and purification on the secondary fraction fine fraction B-2-2-1 to obtain the compound I.
In the step (1), the dried phyllanthus emblica is crushed to 80-100 meshes.
In the step (2), the total extract of the phyllanthus emblica is concentrated in vacuum at the temperature of 50-60 ℃ to obtain the total thick phyllanthus emblica paste.
In the step (3), the total soft extract of the phyllanthus emblica is uniformly suspended in distilled water according to the volume ratio of the total soft extract of the phyllanthus emblica to the distilled water=1:1.5.
And (3) screening the protective activity of different extraction parts by adopting an IFCC method, and determining the n-butanol extraction part as a liver protective active part.
In the step (4), the macroporous adsorption resin is AB-8 type macroporous adsorption resin.
The result shows that the main fraction B has obvious liver protecting activity.
In the step (5), the medium pressure chromatographic column is se:Sup>A YMC-Pack ODS-A column (250X 50mm,5 μm); gradient elution is carried out by taking 15% methanol, 25% methanol and 35% methanol as mobile phases, and absorption wavelength is as follows: 205-230nm, flow rate: 15-20mL/min.
In the step (8), the column for preparing the liquid chromatograph is se:Sup>A YMC-Pack ODS-A column (250X 20mm,5 μm); mobile phase: 10-25% methanol, absorption wavelength: 200-220nm, flow rate: 2-4mL/min.
The inventor adopts a full-automatic biochemical analyzer and an IFCC method to measure ALT and AST activities in serum, and screens and evaluates liver protection activities of a novel compound I, and the result shows that the compound has obvious liver protection activities.
The invention provides application of the lignan compound containing a tannin structure in preparing liver protecting medicines.
The invention provides application of the lignan compound containing a tannin structure in preparing a medicament for treating liver injury.
The invention also provides a pharmaceutical composition with liver protection activity, which takes the lignan compound containing a tannin structure as an active ingredient.
The invention has the beneficial effects that:
the novel lignan compound containing a tannin structure is separated from phyllanthus emblica, and the plant resource is rich and the raw materials are easy to obtain. The compound has simple separation method, easy operation, novel and unique structure, and liver protecting activity, and can be made into liver protecting medicine.
Drawings
FIG. 1 is a scheme for the preparation of Compound I.
Detailed Description
The following describes the essential aspects of the invention in connection with specific examples, but is not intended to limit the scope of the invention.
Example 1
The preparation method of the compound I is shown in figure 1, and comprises the following steps:
crushing dried phyllanthus emblica (8.5 kg) to 90 meshes, soaking the crushed phyllanthus emblica in 90% ethanol for 22 hours, heating and reflux-extracting for 4 times, each time for 5 hours, and combining the filtrates to obtain a phyllanthus emblica total extract;
concentrating the total extract of the phyllanthus emblica in vacuum at the temperature of 55 ℃ to obtain total thick phyllanthus emblica paste (1.1 kg);
step (3), uniformly suspending the total thick phyllanthus emblica paste in distilled water (total thick phyllanthus emblica paste and distilled water=1:1.5, V/V), and sequentially extracting with petroleum ether, ethyl acetate and n-butanol to obtain petroleum ether extraction parts (98.6 g), ethyl acetate extraction parts (207.8 g) and n-butanol extraction parts (554.6 g) respectively;
step (4), screening the different extraction parts obtained in the step (3) for activity protection by adopting an IFCC method, and determining the n-butanol extraction part as a liver protection active part;
step (5), loading the liver-protecting active site obtained in the step (4) on an AB-8 type macroporous adsorption resin column, and sequentially carrying out gradient elution by using 10% ethanol, 25% ethanol, 45% ethanol, 70% ethanol and 100% ethanol to obtain 5 main fractions: a (75.6 g), B (98.0 g), C (60.2 g), D (45.1 g), E (36.7 g);
and (6) screening the liver protection activity of the 5 main fractions obtained in the step (5) by an IFCC method, wherein the result shows that the main fraction B has obvious liver protection activity. The main fraction B was subjected to se:Sup>A medium pressure chromatography column and se:Sup>A YMC-Pack ODS-A column (column length 250X inner diameter 50mm, particle size 5 μm) and eluted with se:Sup>A gradient of 15% methanol, 25% methanol, 35% methanol as mobile phase, with absorption wavelength: 205-230nm, flow rate: 15-20mL/min, 3 secondary fractions were obtained: b-1 (35.7 g), B-2 (26.8 g), B-3 (11.3 g);
step (7), loading the secondary fraction B-2 on a Toyopearl HW-40C column, eluting with methylene dichloride-methanol=1:2 (V/V) to obtain a subfraction B-2-1 (9.6 g) and B-2-2 (13.5 g);
step (8), loading the subfraction B-2-2 on a Sephadex LH-20 column, eluting with 85% methanol to obtain fine fractions B-2-2-1 (4.3 g) and B-2-2-2 (3.1 g);
step (9), carrying out preparative liquid chromatography separation and purification on the fine fraction B-2-2-1, wherein the YMC-Pack ODS-A column (column length 250 multiplied by inner diameter 20mm, granularity 5 μm) is provided with se:Sup>A mobile phase: 20% methanol, absorption wavelength: 210nm, flow rate: 3ml/min, and after multiple preparation and purification, new compound I (12.25 mg) was finally obtained.
Compound I is a white solid, HR-ESI-MS m/z 844.1827[ M+Na ]] + Prompting its molecular formula to be C 40 H 38 O 19 (calcd.for C 40 H 37 O 19 Na, 844.1823). IR v of Compound I max :3291、1701、1618、1378cm -1 ,UV(MeOH)λ max 205, 210, 306, 320nm. In the compound I 1 H NMR 13 In the C NMR spectrum, there are two typical galloyl signals: delta H 7.08 109.1 (C-2 '/6'), 166.2 (C-7 '), 109.1 (C-2'/6 '), 166.2 (C-7'), 146.0 (C-3 '/5'), 138.9 (C-4 ') the galloyl group was linked to the C-3 position of glucose as determined by correlation of H-3 and C-7' in the HMBC spectra. Delta H 7.04 (2H, s, H-2""/6 "") and δc122.1 (C-1 ""), 108.8 (C-2 ""/6 ""), 166.0 (C-7 ""), 145.0 (C-3 ""/5 "").) 138.7 (C-4 '") and the galloyl group was linked to the C-6 position of glucose as determined by correlation of H-6 and C-7' in HMBC spectra. In addition, a typical ABX system signal: delta H 6.53 (1 h, d, j=2.0 hz, h-2 '), 6.50 (1 h, d, j=8.0, 2.0hz, h-6 '), 7.30 (1 h, d, j=8.0 hz, h-5 ') and δc 159.7 (C-1 '), 100.3 (C-2 '), 160.3 (C-3 '), 110.9 (C-4 '), 127.5 (C-5 '), 107.1 (C-6 '), are shown in compound i 1 H NMR 13 In the C NMR spectrum, it was shown that the ABX system was attached at the C-1 'position based on the correlation of H-1 and C-1' in the HMBC spectra. Further analysis found that 1 '-allyloxy-8' -hydroxymethyl-3 "-methoxy-phenylpropanold fragment signal was present 1 H NMR 13 C NMR spectrum: delta H 6.98 (1H, s, H-2 "), 7.26 (1H, s, H-6"), 6.64 (1H, d, J=15.8 Hz, H-7 "), 6.25 (1H, d, J=15.8 Hz, H-8"), 4.19 (2H, m, H-9 "), 4.72 (2H, s, H-9 ') and δc132.1 (C-1"), 105.3 (C-2 "), 144.7 (C-3"), 148.7 (C-4 "), 125.3 (C-5"), 108.9 (C-6 "), 130.2 (C-7"), 127.5 (C-8 "), 64.3 (C-9"), 151.1 (C-7'), 115.2 (C-8 '), 50.0 (C-9'), and correlating the fragment at the C-4 'position according to H-5 and C-7' in the HMBC spectrum. The compound is 1 HNMR(DMSO-d 6 600 MHz) and 13 C NMR(DMSO-d 6 150 MHz) data are shown in table 1. Based on the spectroscopic data of compound I and HMBC, 1 H- 1 H COSY coupling related information, through SciFinder search, compound I is a novel lignan compound containing a tannin structure.
TABLE 1 Compounds I 1 H NMR、 13 C NMR and HMBC related data
Example 2
Liver protection Activity assay of Compound I
1.1 laboratory animals, reagents and instrumentation
SPF-grade male Kunming mice; carbon tetrachloride, silybin, edible peanut oil, glutamic-oxaloacetic transaminase (AST), glutamic-pyruvic transaminase (ALT), CS-600B full-automatic biochemical analyzer, HC-3618R high-speed low-temperature refrigerated centrifuge, ZT-14V2 biological tissue dehydrator, BX51 microscope and the like.
1.2 moulding and administration
SPF-class male Kunming mice were randomly assigned to 6 groups, which were model, control, positive drug, compound I high dose, compound I medium dose, and compound I low dose, respectively. The model group and the control group were perfused with a 0.5% sodium carboxymethyl cellulose solution (dosage 10 mL/kg) daily; the positive control group was administered by gastric lavage with 0.0842g/kg of silybin (0.5% sodium carboxymethylcellulose); the high, medium and low dose groups were each given 2.38g/kg, 1.19g/kg, 0.595g/kg of Compound I (0.5% sodium carboxymethylcellulose solution). Each group was given a continuous dose for 15 days, and after 1h of the last dose, all mice except the control group were given a concentration of 0.15% by intraperitoneal injection of CCl 4 Peanut oil solution (dosage 10 mL/kg) was eaten, then no water was forbidden, after 16h, eyeballs were removed, blood was collected intravenously, and centrifugation was performed at 5000rpm for 15min, and ALT and AST activities in serum were measured using a full-automatic biochemical analyzer and IFCC method.
1.3 Effect of Compound I on ALT and AST Activity in mouse serum
The experimental results are shown in Table 2, and ALT and AST activities in serum of mice in the model group are significantly higher than those in the control group, with significant differences (P<0.01 And) represents CCl 4 The mouse model of the liver injury is successfully established. ALT and AST activities in the serum of mice in the positive control group (silybin) and the three dose group of compound I were reduced compared to the model group; and compared with the model group, ALT and AST activities in serum of mice in the three dose groups of the compound I are significantly reduced (P<0.05 The novel compound I has a certain protection effect on the liver injury of mice, and the compound I shows remarkable liver protection activity onThe novel liver protecting medicine developed from phyllanthus emblica has important scientific value.
TABLE 2 Compound I vs CCl 4 Influence of AST and ALT in serum of mice with liver injury
| Group of | Animal number (only) | Dosage (g/kg) | AST(U/L) | AST(U/L) |
| Model group | 12 | - | 241.84±35.46 | 230.54±41.57 |
| Control group | 12 | - | 114.26±23.89** | 89.53±16.52** |
| Positive pharmaceutical group | 12 | 0.0842 | 202.05±56.17* | 170.32±70.36* |
| High dose group | 12 | 2.38 | 194.28±61.24** | 160±50.52** |
| Medium dose group | 12 | 1.19 | 197.54±42.43* | 163.57±61.19* |
| Low dose group | 12 | 0.595 | 199.76±53.14* | 166.37±56.27* |
Claims (9)
1. Lignan compound containing tannin structure and shown in formula I:
2. the method for producing a tannin-containing lignan compound according to claim 1, characterized by comprising: the method comprises the following steps:
crushing the dried phyllanthus emblica, soaking the crushed phyllanthus emblica in 90-100% ethanol for 20-24 hours, heating and refluxing for 3-5 times, each time for 4-6 hours, and combining the filtrates to obtain a phyllanthus emblica total extract;
step (2), carrying out vacuum concentration on the total extract of the phyllanthus emblica to obtain total thick phyllanthus emblica paste;
uniformly suspending the total phyllanthus emblica thick paste in distilled water, and sequentially extracting with petroleum ether, ethyl acetate and n-butanol to obtain petroleum ether extraction sites, ethyl acetate extraction sites and n-butanol extraction sites respectively;
step (4), the n-butanol extraction part is put on a macroporous adsorption resin column, and gradient elution is carried out by using 10% ethanol, 25% ethanol, 45% ethanol, 70% ethanol and 100% ethanol in sequence, so as to obtain 5 main fractions: main fraction A, main fraction B, main fraction C, main fraction D, main fraction E;
and (5) separating the main fraction B on a medium pressure chromatographic column to obtain 3 secondary fractions: secondary fraction B-1, secondary fraction B-2, secondary fraction B-3;
step (6), loading the secondary fraction B-2 on a Toyopearl HW-40C column, eluting with methylene dichloride-methanol volume ratio=1:2 to obtain a subfraction B-2-1 and a subfraction B-2-2;
step (7), loading the subfraction B-2-2 on a Sephadex LH-20 column, eluting with 70-90% methanol to obtain a fine fraction B-2-2-1 and a fine fraction B-2-2-2;
and (8) carrying out preparation liquid chromatography separation and purification on the secondary fraction fine fraction B-2-2-1 to obtain the compound I.
3. The method for producing a tannin-containing lignan compound according to claim 2, characterized by comprising: in the step (1), the dried phyllanthus emblica is crushed to 80-100 meshes.
4. The method for producing a tannin-containing lignan compound according to claim 2, characterized by comprising: in the step (4), the macroporous adsorption resin is AB-8 type macroporous adsorption resin.
5. The method for producing a tannin-containing lignan compound according to claim 2, characterized by comprising: in the step (5), the medium-pressure chromatographic column is se:Sup>A YMC-Pack ODS-A column: 250X 50mm,5 μm; gradient elution is carried out by taking 15% methanol, 25% methanol and 35% methanol as mobile phases, and absorption wavelength is as follows: 205-230nm, flow rate: 15-20mL/min.
6. The method for producing a tannin-containing lignan compound according to claim 2, characterized by comprising: in the step (8), the column for preparing the liquid chromatograph is se:Sup>A YMC-Pack ODS-A column: 250X 20mm,5 μm; mobile phase: 10-25% methanol, absorption wavelength: 200-220nm, flow rate: 2-4mL/min.
7. The use of a lignan compound having a tannin structure according to claim 1 for preparing a liver protecting agent.
8. The use of a lignan compound having a tannin structure according to claim 1 for preparing a medicament for treating liver injury.
9. A pharmaceutical composition having liver protecting activity, characterized in that: the pharmaceutical composition takes the lignan compound containing a tannin structure as an active ingredient.
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