CN105640933A - Uses of flavonoid compounds in preparation of anti-hepatitis virus medicines - Google Patents
Uses of flavonoid compounds in preparation of anti-hepatitis virus medicines Download PDFInfo
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- CN105640933A CN105640933A CN201410657691.2A CN201410657691A CN105640933A CN 105640933 A CN105640933 A CN 105640933A CN 201410657691 A CN201410657691 A CN 201410657691A CN 105640933 A CN105640933 A CN 105640933A
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Abstract
The invention belongs to the field of medicines and relates to uses of flavonoid compounds in preparation of anti-hepatitis virus medicines. The flavonoid compounds which are 5,7,8-trihydroxy-4'-methoxyflavone (1) and potengriffioside A (2) are extracted and separated from helicteres angustifolia. Tests prove that the flavonoid compounds have anti-hepatitis virus activity. Pharmacological test results with a lamivudine positive control show that the flavonoid compounds have obvious anti-hepatitis virus functions, wherein the IC50 value for HBsAg of the 5,7,8-trihydroxy-4'-methoxyflavone (1) is 49.3 mug/mL, the IC50 value for HBeAg of the 5,7,8-trihydroxy-4'-methoxyflavone (1) is 68.4 mug/mL, the IC50 value for HBsAg of the potengriffioside A (2) is 14.6 mug/mL, the IC50 value for HBeAg of the potengriffioside A (2) is 41.5 mug/mL, the above mentioned IC50 values are obviously higher than those of the lamivudine positive control. The flavonoid compounds can be adopted as active components and used for preparing the anti-hepatitis virus medicines.
Description
Technical field
The invention belongs to field of medicaments, it relates to from medicinal plant Root of Narrowleaf Screwtree, be separated the flavonoid compound obtained in the purposes preparing in anti-hepatic-B virus medicine.
Background technology
Showing according to the relevent statistics, artificial chronic viral hepatitis B virus (HBV) carrier in the whole world nearly more than 300,000,000, China accounts for half. Hepatitis B virus infection has become one of global main disease affecting human health, owing to HBV infection can cause chronic hepatitis, liver cirrhosis and primary hepatocarcinoma, therefore becomes one of nine big diseases affecting human longevity. Owing to chronic HBV infection person occurs the relative risk rate of liver cancer to be 217 compared with noncarrier, therefore in the whole world, nearly 2,000,000 people die from liver cirrhosis and the liver cancer that hepatitis B causes every year, and China is the district occurred frequently of hepatitis B, 250,000 people are about had to die from the chronic hepatopathy (liver cirrhosis and liver cancer) relevant to hepatitis B every year. Above-mentioned disease brings massive losses to people ' s health and national economy. Therefore, the Anti-HBV drugs seeking high-efficiency low-toxicity has become very urgent problem.
Pharmaceutical chemists has carried out unremitting effort in the process of screening Anti-HBV drugs, but the specifics that up to the present can cure HBV infection is also being found. The alpha-interferon and the nucleoside medicine lamivudine that have obtained FDA official approval use on clinical can only obtain result for the treatment of to a certain extent, most patient is not also reached the object of healing, stopping " knock-on " phenomenon after treatment is that medical personal feels stubborn problem most, so the Anti-HBV activity new drug developing poison efficient, low, not " knock-on " proposes challenge to pharmaceutical chemists. In the process of screening Anti-HBV activity new drug, more and more pharmaceutical chemists has turned to attention that plant origin is abundant, cheap and easy to get, toxic side effect is little, the complicated and diversified natural compounds of molecular structure.
Root of Narrowleaf Screwtree (HelicteresangustifoliaL.) is Sterculiaceae plant, mainly it is distributed in Lingnan area, its root or complete stool are medicinal, there is the function of heat-clearing of inducing sweat, subduing swelling and detoxicating, it is usually used in the diseases such as treatment cold, fever, cough due to lung-heat, swelling and pain in the throat, headache, enteritis, carbuncle be swollen, also often occurs in formula of herbal tea. Pharmacological research shows that Root of Narrowleaf Screwtree has inflammation antibacterial, anti-, antiviral and antitumor isoreactivity. Our research shows that Root of Narrowleaf Screwtree crude extract has good Anti-HBV effect, but the effective substance of its Anti-HBV activity is lacked research, and the flavonoid compound that the present invention relates to there is not yet Anti-HBV effect report.
Summary of the invention
It is an object of the invention to provide the flavonoid compound being separated from Root of Narrowleaf Screwtree and obtaining in the purposes preparing in anti-hepatic-B virus medicine.
The present invention's application modern pharmacology screening method, belong to extraction and isolation (Helicteres) plant Root of Narrowleaf Screwtree (H.angustifolia) from Sterculiaceae Root of Narrowleaf Screwtree and obtain flavonoid compound: 5,7,8-trihydroxy--4'-methoxy flavone (1) and potengriffioside A (2), and confirm that described compound has Anti-HBV effect through test.
Active compound of the present invention has following chemical structure:
In the present invention,
Compound 5,7,8-trihydroxy--4'-methoxy flavone (1), wherein, R1=OH, R2=H, R3=OCH3;
Compound potengriffioside A (2), wherein, R1=H, R2=OA, R3=OH, and the group A in substituent R 2 is:
The compound of the present invention is prepared by following method:
Get Root of Narrowleaf Screwtree dry aerial parts 15kg, 95% alcohol reflux, respectively with sherwood oil, ethyl acetate and n-butanol extraction after concentrated, it is total to obtain 1.05kg, get acetic acid ethyl ester extract 160g and carry out silica gel column chromatography, with methylene chloride-methanol gradient elution, gained stream part carries out repeatedly silica gel column chromatography, MPLC with SephadexLH-20 gel column chromatography etc. with different eluent and be separated means of purification and be separated and obtain compound 5,7,8-trihydroxy--4'-methoxy flavone (1) and potengriffioside A (2).
Flavonoid compound 5 of the present invention, 7,8-trihydroxy--4'-methoxy flavone (1) and potengriffioside A (2) have carried out the experiment of external Anti-HBV activity, result confirms, described compound has significant Anti-HBV effect, and effective concentration is low, and wherein 5,7,8-trihydroxy--4'-methoxy flavone (1) is to the IC of HBsAg50Value is 49.3 �� g/ml, to the IC of HBeAg50Value is 68.4 �� g/ml; Potengriffioside A (2) is to the IC of HBsAg50Value is 14.6 �� g/ml, to the IC of HBeAg50Value is 41.5 �� g/ml, is significantly higher than positive control lamivudine; Described compound can be used as the medicine of activeconstituents for the preparation of Anti-HBV activity.
Embodiment
Embodiment 1 extracting and developing, obtained flavonoid compound
Get Root of Narrowleaf Screwtree dry root 15kg, pulverize, with 95% alcohol reflux 3 times (50L �� 3), each 2h, united extraction liquid also concentrated to obtain medicinal extract 1.05kg, and add water (4L) is mixed outstanding, respectively with equal-volume sherwood oil, ethyl acetate and n-butanol extraction 5 times, combining extraction liquid also concentrates to dry, obtains acetic acid ethyl ester extract 160g. By Ethyl acetate fraction through silica gel (200-300 order) pillar layer separation, successively with methylene chloride-methanol (50:1-0:1) gradient elution, obtain 11 streams part (Fr.1-11), wherein flow part Fr.3 (30g) again through silica gel column chromatography (methylene chloride-methanol, 30:1,20:1,10:1,5:1,3:1,1:1), MPLC (methanol-water, 20:80-80:20 gradient elution) and the means purifying such as SephadexLH-20 gel column chromatography, separation obtains compound 5,7,8-trihydroxy--4'-methoxy flavone (1); Stream part Fr.4 (25g) is again through silica gel column chromatography (methylene chloride-methanol, 30:1,20:1,10:1,5:1,3:1,1:1), MPLC (methanol-water, 20:80-80:20 gradient elution) and the means purifying such as SephadexLH-20 gel column chromatography, separation obtains compound potengriffioside A (2).
Wherein, 5,7,8-trihydroxy--4'-methoxy flavone (5,7,8-trihydroxy-4'-methoxyflavone, 1) for brown color without sizing powder, UV (MeOH) ��maxNm (log ��): 360 (2.54), 324 (3.27), 300 (2.86), 272 (3.11);IR (KBr): ��max(cm-1): 3380,1664,1616,1582; ESI-MSm/z:301 [M+H]+��1H-NMR(400MHz,DMSO-d6)��H: 6.81 (1H, s, H-3), 6.27 (1H, s, H-6), 8.11 (2H, d, J=8.0Hz, H-2', 6'), 7.11 (2H, d, J=8.0Hz, H-3', 5'), 12.34 (1H, s, 5-OH), 3.85 (3H, s, 4'-OCH3);13C-NMR(100MHz,DMSO-d6)��C:56.2(-OCH3),99.1(C-6),103.4(C-3),103.8(C-10),114.5(C-3',C-5'),121.2(C-8),122.7(C-1'),128.6(C-2',C-6'),149.5(C-9),156.8(C-5),157.2(C-7),162.4(C-4'),163.3(C-2),182.1(C-4)��
Potengriffioside A (potengriffiosideA, 2) is the amorphous powder of yellow,(c0.25, MeOH); UV (MeOH) ��maxNm (log ��): 360 (3.21), 315 (3.47), 302 (2.95), 276 (3.43); IR (KBr): ��max(cm-1): 3385,1660,1606,1512,1448,1366,1167,1077,844; ESI-MSm/z:595 [M+H]+��1H-NMR(400MHz,DMSO-d6)��H: 8.07 (2H, d, J=9.0Hz, H-2', 6'), 7.49 (1H, d, J=15.6Hz, H-7 " '), 7.38 (2H, d, J=8.4Hz, H-2 " ', 6 " '), 6.90 (2H, d, J=9.0Hz, H-3', 5'), 6.88 (2H, d, J=9.0Hz, H-3 " ', 5 " '), 6.36 (1H, d, J=2.4Hz, H-8), 6.20 (1H, d, J=2.4Hz, H-6), 6.17 (1H, d, J=15.6Hz, H-8 " '), 5.34 (1H, m, J=4.2Hz, H-1 "), 4.41 (1H, dd, J=12.0, 11.4Hz, H-6 is " b), 4.30 (1H, dd, J=11.4, 12.0Hz, H-6 is " a),13C-NMR(100MHz,DMSO-d6)��C:158.6(C-2),135.5(C-3),179.6(C-4),163.2(C-5),100.2(C-6),166.1(C-7),95.1(C-8),159.6(C-9),105.9(C-10),123.0(C-1'),132.5(C-2',6'),116.3(C-3',5'),161.4(C-4'),104.3(C-1��),76.1(C-2��),78.3(C-3��),72.0(C-4��),76.0(C-5��),64.7(C-6��),127.3(C-1��'),131.5(C-2��',6��'),117.0(C-3��',5��'),161.8(C-4��'),146.8(C-7��'),115.0(C-8��'),169.1(C-9��')��
The external Anti-HBV activity experiment of embodiment 2
The 2.2.15 cell strain (the Ministry of Education/medical molecular virology key lab of the Ministry of Health, Shanghai) of application HepG2, with every hole 10 �� 105Individual cell is inoculated in 24 orifice plates, and substratum is DMEM, and growth media is containing 10% foetal calf serum, and 380 �� g/mlG418,0.03% glutamines, each 100 �� g/ml of penicillin, Streptomycin sulphate, at 5%CO2Incubate 37 DEG C of cultivations in case, after 48 hours, change the nutrient solution that dimethyl sulfoxide (DMSO) helps molten drug containing into, 3��5 concentration established by often kind of medicine, each concentration establishes 4 parallel holes, continues to cultivate 9 days (changing liquid once in every 3 days), collects supernatant liquor ELISA and detects HBsAg and HBeAg content. Under same condition with the nutrient solution supernatant liquor of not drug containing as a control group. Use above-mentioned cell strain simultaneously, measure the cytotoxicity of medicine with mtt assay. Positive control is lamivudine (3TC). Result confirms that compound of the present invention has significant Anti-HBV effect, and effective concentration is low, and wherein 5,7,8-trihydroxy--4'-methoxy flavone (1) is to the IC of HBsAg50Value is 49.3 �� g/ml, to the IC of HBeAg50Value is 68.4 �� g/ml; Potengriffioside A (2) is to the IC of HBsAg50Value is 14.6 �� g/ml, to the IC of HBeAg50Value is 41.5 �� g/ml, is significantly higher than positive control lamivudine (table 1).
The restraining effect that HBsAg and HBeAg is secreted by table 1. compound 1 and 2
/: inhibiting rate test is not carried out when cytotoxicity >=25%
The reagent testing employing in the present invention is techniques well known, commercially available.
Claims (2)
1. the flavonoid compound with having structure in the purposes preparing in anti-hepatic-B virus medicine,
Described compound is 5,7,8-trihydroxy--4'-methoxy flavone (1) and potengriffioside A (2),
Wherein,
Compound 5,7,8-trihydroxy--4'-methoxy flavone (1), wherein, R1=OH, R2=H, R3=OCH3;
Compound potengriffioside A (2), wherein, R1=H, R2=OA, R3=OH, and the group A in substituent R 2 is:
2. by purposes according to claim 1, it is characterized in that, described compound is prepared by following method: get Root of Narrowleaf Screwtree dry root, pulverizes, with 95% alcohol reflux 3 times, each 2h, united extraction liquid also concentrated to obtain medicinal extract, adds water mixed outstanding, respectively with equal-volume sherwood oil, ethyl acetate and n-butanol extraction 5 times, combining extraction liquid also concentrates to dry, obtains acetic acid ethyl ester extract;
By Ethyl acetate fraction through silica gel 200-300 order pillar layer separation, successively with methylene chloride-methanol 50:1-0:1 gradient elution, obtain 11 stream part Fr.1-11, wherein flow part Fr.3 again through silica gel column chromatography (methylene chloride-methanol, 30:1,20:1,10:1,5:1,3:1,1:1), MPLC (methanol-water, 20:80-80:20 gradient elution) and SephadexLH-20 gel column chromatography purifying, separation obtains compound 5,7,8-trihydroxy--4'-methoxy flavone (1); Stream part Fr.4 is again through silica gel column chromatography (methylene chloride-methanol, 30:1,20:1,10:1,5:1,3:1,1:1), MPLC (methanol-water, 20:80-80:20 gradient elution) and SephadexLH-20 gel column chromatography purifying, separation obtains compound potengriffioside A (2).
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020053249A1 (en) * | 2018-09-14 | 2020-03-19 | F. Hoffmann-La Roche Ag | Flavone compounds for the treatment and prophylaxis of hepatitis b virus disease |
| CN111265539A (en) * | 2020-03-19 | 2020-06-12 | 暨南大学 | Application of tiliroside in preparation of anti-influenza drugs |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001247470A (en) * | 2000-03-03 | 2001-09-11 | Morishita Jintan Kk | Agent for protecting liver |
-
2014
- 2014-11-18 CN CN201410657691.2A patent/CN105640933A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001247470A (en) * | 2000-03-03 | 2001-09-11 | Morishita Jintan Kk | Agent for protecting liver |
Non-Patent Citations (5)
| Title |
|---|
| AMEHA SEYOUM等: "Structure–radical scavenging activity relationships of flavonoids", 《PHYTOCHEMISTRY》 * |
| ARACELI SALA等: "Assessment of the anti-inflammatory activity and free radical scavenger activity of tiliroside", 《EUROPEAN JOURNAL OF PHARMACOLOGY》 * |
| FRANCESCA DANESI等: "Bioactive-rich Sideritis scardica tea (mountain tea) is as potent as Camellia sinensis tea at inducing cellular antioxidant defences and preventing oxidative stress", 《J SCI FOOD AGRIC》 * |
| MIYAHARA M等: "Structure-activity relationship of flavonoids in suppressing rat liver lipid peroxidation", 《JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN》 * |
| 张冠群 等: "乙型肝炎与脂质过氧化作用的研究", 《广东医学》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020053249A1 (en) * | 2018-09-14 | 2020-03-19 | F. Hoffmann-La Roche Ag | Flavone compounds for the treatment and prophylaxis of hepatitis b virus disease |
| WO2020052774A1 (en) * | 2018-09-14 | 2020-03-19 | F. Hoffmann-La Roche Ag | Flavone derivatives for the treatment and prophylaxis of hepatitis b virus disease |
| CN112601743A (en) * | 2018-09-14 | 2021-04-02 | 豪夫迈·罗氏有限公司 | Flavone compounds for treating and preventing hepatitis B virus diseases |
| CN112601743B (en) * | 2018-09-14 | 2023-12-19 | 豪夫迈·罗氏有限公司 | Flavone compounds for the treatment and prevention of hepatitis b virus diseases |
| US12012390B2 (en) | 2018-09-14 | 2024-06-18 | Hoffmann-La Roche Inc. | Flavone compounds for the treatment and prophylaxis of hepatitis B virus disease |
| CN111265539A (en) * | 2020-03-19 | 2020-06-12 | 暨南大学 | Application of tiliroside in preparation of anti-influenza drugs |
| CN111265539B (en) * | 2020-03-19 | 2021-05-18 | 暨南大学 | Application of tiliroside in preparation of anti-influenza drugs |
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