CN1319531C - Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine - Google Patents

Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine Download PDF

Info

Publication number
CN1319531C
CN1319531C CNB2005100265732A CN200510026573A CN1319531C CN 1319531 C CN1319531 C CN 1319531C CN B2005100265732 A CNB2005100265732 A CN B2005100265732A CN 200510026573 A CN200510026573 A CN 200510026573A CN 1319531 C CN1319531 C CN 1319531C
Authority
CN
China
Prior art keywords
alkaloid
hepatitis
preparation
methoxydicamnine
fagarine
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
CNB2005100265732A
Other languages
Chinese (zh)
Other versions
CN1698610A (en
Inventor
陈道峰
杨国红
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fudan University
Original Assignee
Fudan University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fudan University filed Critical Fudan University
Priority to CNB2005100265732A priority Critical patent/CN1319531C/en
Publication of CN1698610A publication Critical patent/CN1698610A/en
Application granted granted Critical
Publication of CN1319531C publication Critical patent/CN1319531C/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The present invention relates to the new purpose of quinoline alkaloid in the preparation of a drug for resisting hepatitis B viruses, particularly to the purpose of skimmianine and 5-methoxydictamnine in the preparation of a drug for treating hepatitis B and an extraction method thereof, which belongs to the field of traditional Chinese medicine pharmacy. In the present invention, active alkaloid, quinoline alkaloid, extracted from the alkaloid position of rutaceae zanthoxylum nitidum has the obvious action of resisting HBV through the verification of the result of a pharmacological test and has the characteristics of low effective concentration and less cytotoxicity. The measured alkaloid displays the inhibiting action on HBsAg and HBeAg. The inhibition ratio of the 5-methoxydictamnine is higher than that of positive control lamivudine, and the inhibiting action of skimmianine on the HBV is equivalent to that of the lamivudine.

Description

The purposes of quinoline alkaloid in the preparation anti-hepatic-B virus medicine
Technical field
The invention belongs to field of traditional Chinese medicine pharmacy, relate to the new purposes of quinoline alkaloid in the preparation anti-hepatic-B virus medicine.Especially .beta.-fagarine and the application of 5-methoxydicamnine in anti-hepatic-B virus medicine.
Background technology
It is global one of the principal disease of human health that influences that hepatitis B virus (HBV) infects, according to statistics, the whole world nearly more than 300,000,000 artificial Chronic HBV carriers, China accounts for half.Can cause chronic hepatitis, liver cirrhosis and primary hepatocarcinoma because HBV infects, therefore become one of nine big diseases that influence human longevity.Because the relative risk rate of the more non-carrier generation of chronic HBV infection person hepatocarcinoma is 217, therefore annual nearly 2,000,000 people die from liver cirrhosis and the hepatocarcinoma that hepatitis B causes in the whole world, and China is the district occurred frequently of hepatitis B, and annual have 250,000 people to die from the chronic hepatopathy relevant with hepatitis B (liver cirrhosis and hepatocarcinoma) approximately.This brings massive losses for people ' s health and national economy.Therefore, the anti-HBV medicine of seeking high-efficiency low-toxicity has become instant problem.
Although pharmaceutical chemists has been carried out unremitting effort in the process of the anti-HBV medicine of screening, up to the present people also do not find a kind of specific drug to cure the HBV infection.The alpha-interferon and the nucleoside medicine lamivudine that have obtained FDA official approval use clinically can only obtain therapeutic effect to a certain extent, most patient is not also reached the purpose of healing, " knock-on " phenomenon after stopping to treat is that medical worker feels stubborn problem most, so develop efficient, low toxicity, the anti-HBV new drug of " knock-on " has not proposed challenge to pharmaceutical chemists.In the process of the anti-HBV new drug of screening, increasing pharmaceutical chemists has turned to attention plant origin abundant, and cheap and easy to get, toxic and side effects is little, the complicated and diversified native compound of molecular structure.
Chinese medicine Radix Zanthoxyli (Zanthoxylum nitidum) belongs to Rutaceae Pericarpium Zanthoxyli plant, mainly is distributed in provinces and regions, China south, and as Guangxi, Guangdong, Fujian and other places, the Guangxi resource is especially abundant.Its root commonly used among the people, rhizome, root bark and peel of stem are used as medicine, and have the effect that promoting the circulation of QI to relieve pain, promoting blood circulation to remove blood stasis, collateral dredging are dispeled the wind, and are used for traumatic injury, rheumatic arthralgia, toothache stomachache, venom.Being applied to clean the teeth, is the primary raw material of making shiny pricklyash toothpaste; In recent years, discover that it has anti-tumor activity.This medical material contains benzene a pair of horses going side by side aza-phenanthrenes Alkaloid, quinoline alkaloid and Ingredients Such As Coumarin, and wherein benzene a pair of horses going side by side aza-phenanthrenes Alkaloid is the antitumor effective substance of this medical material.But do not see the effect of the antagonism of quinoline alkaloid monomer in Radix Zanthoxyli HBV so far as yet and be used for the treatment of the report of hepatitis B.
Summary of the invention
The purpose of this invention is to provide new antiviral active substance, be specifically related to the new purposes of quinoline alkaloid in the preparation anti-hepatic-B virus medicine.Be particularly related to .beta.-fagarine and 5-methoxydicamnine purposes and the extracting method thereof in preparation treatment hepatitis B medicine.
The present invention is according to " eliminating pathogenic factor for supporting vital QI " principle, use the modern pharmacology screening technique, the active substance that suppresses HBV in the plant amedica is studied, and the alkaloid position is extracted the active alkaloid quinoline alkaloid and is confirmed that it has the resisting HBV virus activity from Rutaceae xanthoxylum Radix Zanthoxyli (Zanthoxylumnitidum).
Quinoline alkaloid of the present invention comprises .beta.-fagarine (1) and 5-methoxydicamnine (2).
Active alkaloid compounds of the present invention has the chemical constitution of formula I:
Wherein: 1 R 1=H, R 2=R 3=OCH 3,
2 R 1＝OCH 3，R 2＝R 3＝H。
Chemical compound quinoline alkaloid of the present invention prepares by following method:
Get Radix Zanthoxyli rhizome coarse powder 10kg, be not the alkaloid reaction with 2% sulphuric acid (v/v) room temperature merceration (30L * 4 time) to acid liquid, merge acid water extracting liquid, alkalizing through NaOH repeatedly extracts fully with the chloroform equal-volume to pH10～11, and the reclaim under reduced pressure chloroform gets total extract 110g (extraction ratio 1.10%).Sample carries out silica gel column chromatography on the total alkali extractum 100g dry method, with chloroform, chloroform-methanol, methanol gradient elution, the gained flow point carries out repeatedly silica gel column chromatography and sephadex LH-20 column chromatography with different eluant, separates to obtain .beta.-fagarine (1), 5-methoxydicamnine (2).
Wherein .beta.-fagarine (skimmianine) (1) white plates crystallization (methanol), mp 168-170 ℃; C 14H 13NO 4, molecular weight 259; 1HNMR (CDCl 3, 400MHz) δ ppm:8.00 (1H, d, J=9.39Hz, 5-H), 7.57 (1H, d, J=2.74Hz, 2-H), 7.22 (1H, d, J=9.39Hz, 6-H), 7.02 (1H, d, J=2.74Hz, 3-H), 4.42 (3H, s, 4-OCH 3), 4.11 (3H, s, OCH 3), 4.02 (3H, s, OCH 3); 13C-NMR (CDCl 3, 100MHz) δ ppm:164.3 (C-9 '), 157.2 (C-4), 152.1 (C-8 '), 143.0 (C-2), 141.8 (C-7), 141.4 (C-8), 118.2 (C-5), 114.8 (C-3 '), 111.9 (C-6), 104.6 (C-3), 101.9 (C-4 '), 61.6 (8-OCH 3), 58.9 (4-OCH 3), 56.7 (7-OCH 3); EI-MS m/z:259 ([M] +) (70), 258 (26), 244 (100), 230 (53), 229 (26), 216 (24), 213 (26), 201 (26).
Wherein, the white needle of 5-methoxydicamnine (5-methoxydictamnine) (2) (acetone-petroleum ether), mp 126-128 ℃; C 13H 11NO 3, molecular weight is 229; IR (KBr) υ Max(cm -1): 2952,1629,1618,1580,1552,1516,1464,1446,1393,1365,1329,1303,1260,1188,1156,1047,979,812,770,747,725,641; 1HNMR (CDCl 3, 400MHz) δ ppm:7.83 (1H, d, J=8.61Hz, 8-H), 7.63 (1H, d, J=2.74Hz, 2-H), 7.35 (1H, t, J=8.61Hz, 7-H), 7.05 (1H, d, J=8.61Hz, 6-H), 7.06 (1H, d, J=2.74Hz, 3-H), 4.43 (3H, s, 4-OCH 3), 4.07 (3H, s, 5-OCH 3); 13C-NMR (CDCl 3, 100MHz) δ ppm:163.1 (C-9 '), 156.8 (C-4), 154.5 (C-5), 143.9 (C-2), (137.4 C-8 '), 123.4 (C-7), 119.6 (C-4 '), 114.0 (C-8), 107.6 (C-6), 104.5 (C-3), 103.8 (C-3 '), 59.0 (8-OCH 3), 55.9 (4-OCH 3); EI-MS m/z:229 ([M] +) (100), 228 (72), 214 (29), 200 (65), 199 (30), 185 (17), 184 (24), 156 (25).
Above-mentioned alkaloid is through pharmacological testing, and the result confirms to have the significant anti-HBV effect, and valid density is low, cytotoxicity features of smaller (table 1).The alkaloid of surveying HBsAg and HBeAg are all shown inhibitory action.Described 5-methoxydicamnine suppression ratio to HBsAg when 0.2 μ mol/mL is higher than positive control lamivudine (suppression ratio is 29.6% during 1.0 μ mol/mL).Described .beta.-fagarine also has certain inhibitory action to HBeAg when 0.2 μ mol/mL, suppression ratio is 31.9% and lamivudine quite (during 1.0 μ mol/mL is 35.4%).5-methoxydicamnine suppression ratio to HBeAg when 0.2 μ mol/mL is higher than positive control lamivudine (suppression ratio is 35.4% during 1.0 μ mol/mL).
Description of drawings:
Fig. 1 is chloroform extraction position, a Radix Zanthoxyli rhizome acid extraction alkalization back extraction separation flow chart.
The specific embodiment
Embodiment 1 preparation quinoline alkaloid
Radix Zanthoxyli rhizome coarse powder 10kg, be not the alkaloid reaction with 2% sulphuric acid (v/v) room temperature merceration (30L * 4 time) to acid liquid, merge acid water extracting liquid, alkalizing through NaOH repeatedly extracts fully with the chloroform equal-volume to pH10～11, and the reclaim under reduced pressure chloroform gets total extract 110g (extraction ratio 1.10%).Sample carries out silica gel column chromatography on the total alkali extractum 100g dry method, and with chloroform, chloroform-methanol, methanol gradient elution, further concrete operations are as follows:
1. chloroform-methanol (98: 2) eluting gained stream part is carried out silica gel column chromatography, petroleum ether-ethyl acetate (10: 1～6: 4) eluting.Petroleum ether-ethyl acetate (7: 3) stream part, silica gel column chromatography gets white plates crystallization .beta.-fagarine (1) 8mg through Sephedax LH-20 post (80% methanol) more repeatedly.
2. stream part of chloroform-methanol (95: 5) eluting gained is carried out silica gel column chromatography, petroleum ether-ethyl acetate (7: 3) eluting.Petroleum ether-ethyl acetate (6: 4) stream part, silica gel column chromatography gets white, needle-shaped crystals 5-methoxydicamnine (2) 10mg through sephedax LH-20 post (80% methanol) more repeatedly.
Embodiment 2 external anti-HBV tests
HepG2 2.2.1.5 (Ministry of Education/Ministry of Public Health medical molecular virology key lab, Shanghai) after one week of cultivation, adds the sample liquid of 0.2 μ mol/ml in 96 porocyte culture plates, and 37 ℃, 5%CO 2Cultivate 16h, with the toxicity of mtt assay detection compound to HepG2 2.2.1.5 cell, microplate reader reads OD570 (nm) value.
HepG2 2.2.1.5 cultivates a week in 96 porocyte culture plates after, add the sample liquid of 0.2 μ mol/ml, 37 ℃, 5%CO 2Cultivate 24h, collect supernatant, detect HBV antigen with the ELISA method and change.Concrete operations are as follows: HepG2 2.2.1.5 supernatant 10 μ l and 5% confining liquid, 40 μ l are added 96 orifice plates, two multiple hole; Every hole adds 50 μ l enzyme conjugates, mixing, 37 ℃ of water-bath 30min; Abandon liquid, every hole adds 200 μ l PBST, leaves standstill 20 seconds, abandons liquid again, repeats five times; Add 50 μ l colour developing liquid A and 50 μ l colour developing liquid B, mixing, 37 ℃ of water-bath 15min; Every hole adds stop buffer 50 μ l; Microplate reader reads OD450 (nm) value, reference wavelength 650 (nm).
Table 1 is an alkaloid of the present invention to the toxicity of HepG2 2.2.15 cell and to the inhibition effect of HBsAg and HBeAg.
Table 1
Chemical compound (concentration 0.2 μ mol/ml) Cytotoxicity To HBsAg suppression ratio % To HBeAg suppression ratio %
1 2 3TC(1.0μmol/ml) - - / 49.3 29.6 31.9 43.2 35.4
Wherein ,-: " not seeing overt toxicity " means with mtt assay and detects cell survival rate 〉=75%; +: " demonstration toxicity " phalangeal cell survival rate＜75%.

Claims (2)

1, the quinoline alkaloid .beta.-fagarine of formula I structure or the 5-methoxydicamnine purposes in preparation inhibition hepatitis B virus medicine,
1 R 1＝H R 2＝R 3＝OCH 3
2 R 1＝OCH 3 R 2＝R 3＝H。
2, purposes according to claim 1, wherein said quinoline alkaloid .beta.-fagarine or 5-methoxydicamnine prepare by following step:
(1) the Radix Zanthoxyli rhizome is with 2% sulphuric acid (v/v) room temperature merceration, and 30L * 4 time are not the alkaloid reaction to acid liquid, merge acid water extracting liquid, and alkalizing through NaOH repeatedly extracts fully with the chloroform equal-volume to pH10～11, and the reclaim under reduced pressure chloroform gets biology total alkali;
(2) the Radix Zanthoxyli biology total alkali obtains quinoline alkaloid .beta.-fagarine (1), 5-methoxydicamnine (2) through silica gel column chromatography and sephadex LH-20 column chromatography repeatedly.
CNB2005100265732A 2005-06-08 2005-06-08 Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine Expired - Fee Related CN1319531C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNB2005100265732A CN1319531C (en) 2005-06-08 2005-06-08 Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB2005100265732A CN1319531C (en) 2005-06-08 2005-06-08 Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine

Publications (2)

Publication Number Publication Date
CN1698610A CN1698610A (en) 2005-11-23
CN1319531C true CN1319531C (en) 2007-06-06

Family

ID=35474961

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB2005100265732A Expired - Fee Related CN1319531C (en) 2005-06-08 2005-06-08 Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine

Country Status (1)

Country Link
CN (1) CN1319531C (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103041015B (en) * 2012-12-29 2014-09-10 南宁多灵生物科技有限公司 Medicine for treatment of hepatitis

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4767626A (en) * 1985-03-11 1988-08-30 Theodore Cheng Remedy for anemia and arthritis
CN1399558A (en) * 1999-08-23 2003-02-26 免疫平衡技术有限公司 Treatment of viral infections
US6790961B2 (en) * 1992-08-28 2004-09-14 3M Innovative Properties Company Process for preparing 1-substituted, 2-substituted 1H-imidazo[4,5-C]quinolin-4-amines
CN1535151A (en) * 2001-06-07 2004-10-06 ʷ��˿�������ȳ�ķ���޹�˾ Novel anti-infectives

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4767626A (en) * 1985-03-11 1988-08-30 Theodore Cheng Remedy for anemia and arthritis
US6790961B2 (en) * 1992-08-28 2004-09-14 3M Innovative Properties Company Process for preparing 1-substituted, 2-substituted 1H-imidazo[4,5-C]quinolin-4-amines
CN1399558A (en) * 1999-08-23 2003-02-26 免疫平衡技术有限公司 Treatment of viral infections
CN1535151A (en) * 2001-06-07 2004-10-06 ʷ��˿�������ȳ�ķ���޹�˾ Novel anti-infectives

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
生物碱的抗HIV逆转酶活性 中草药，第28卷第10期 1997 *

Also Published As

Publication number Publication date
CN1698610A (en) 2005-11-23

Similar Documents

Publication Publication Date Title
Li et al. Anti-hepatitis B virus activities of Geranium carolinianum L. extracts and identification of the active components
Geng et al. Three new anti-HBV active constituents from the traditional Chinese herb of Yin-Chen (Artemisia scoparia)
CN102861112B (en) Kosteletzkya virginica lignan extract and preparation method and application thereof
WO2008138243A1 (en) A preparation method of icaritin
CN102302685B (en) Common lophatherum herb extract and preparation method and application thereof
CN101759544A (en) Novel chalcone compound and preparation method and application thereof
CN101240005A (en) Method for preparing platycodin D from balloon-flower root and application thereof in anti-cancer medicament
CN101328114B (en) Method for extracting walnuts ketone from walnut green husk
CN105920064A (en) Natural active ingredient extracted and separated from leaves and stems of panax quinuefolium L and application of natural active ingredient
CN106008543A (en) Novel diterpenoid compound and preparation method thereof
CN105037464A (en) Plant flavone compounds, and preparation method and application thereof
CN104873560A (en) Plant general flavone and preparation method and application thereof
CN1319531C (en) Application of quinolines alkaloid in preparation of hepatitis B virus resisting medicine
CN102600268A (en) Gallbladder wood leaf composition containing strictosamide and preparation thereof
CN103784427B (en) Containing the pharmaceutical composition of eudesmane type sesquiterpene and the application in pharmacy thereof
CN1698606A (en) Application of quinolizidine kind alkaloid in preparation of hepatitis B virus resisting medicine
CN101375841B (en) Daucane type sesquiterpenes and preparation method and application thereof
CN101375842B (en) Application of lignan of biphenyl cyclooctene series in preparing anti-hepatitis B virus medicament
CN102603522B (en) Phenol derivatives and application thereof in preparation of anti-HBV (hepatitis B virus) medicines
CN105198943A (en) Acylation flavone glycoside named camellikaempferoside A and preparing method and application thereof
CN104523733A (en) Pharmaceutical composition with anti-aging effect
CN114634537A (en) Preparation method and application of diterpene in gold leaves
CN100341888C (en) Anticancer new compound of Xiacaogan I, preparation method, and application in pharmacy
CN105640933A (en) Uses of flavonoid compounds in preparation of anti-hepatitis virus medicines
CN105663110A (en) Application of lignan compound 4-keto pinoresinol in preparing anti-hepatitis B virus medicine

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20070606

Termination date: 20140608

EXPY Termination of patent right or utility model