CN101787004A - Lignanoid compound contained in Yunnan daphne herb, as well as preparation method and application thereof - Google Patents
Lignanoid compound contained in Yunnan daphne herb, as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN101787004A CN101787004A CN 201010118201 CN201010118201A CN101787004A CN 101787004 A CN101787004 A CN 101787004A CN 201010118201 CN201010118201 CN 201010118201 CN 201010118201 A CN201010118201 A CN 201010118201A CN 101787004 A CN101787004 A CN 101787004A
- Authority
- CN
- China
- Prior art keywords
- compound
- daphne
- hiv
- preparation
- yunnan
- 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.)
- Granted
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a new lignanoid compound (I) contained in Yunnan daphne herb, as well as a preparation method and an application thereof. The preparation method comprises the following steps of: after pulverizing a Yunnan daphne herb sample, ultrasonically extracting the sample with methanol three times, and merging the extracting liquids; filtering and concentrating the extracting liquid in vacuum to a small volume; after settlement, filtering to remove deposits, and concentrating into an extract; preliminarily separating the extract through silica gel column chromatography; and finally, further separating the extract through highly efficient semipreparative chromatogram to obtain the new compound. The compound is screened for antioxidant activity and anti-HIV-1 activity, and test results show that the compound has high antioxidant activity and anti-HIV-1 activity.
Description
Technical field
The present invention relates to extraction separation compound from natural phant, more particularly, the present invention relates to contained Lignanoids compounds of a kind of new Shortpetal Daphne and its production and application.
Background technology
Shortpetal Daphne (Daphne acutiloba Rehd.) is the complete stool of thymelaeceae plant Shortpetal Daphne, and English name Shortpetal Daphne is distributed in ground such as Yunnan, Guizhou, Sichuan.Shortpetal Daphne is except that as the ornamental plant, and pharmaceutical use is also very high.Root of Winter Daphne, bark, Hua Junke are used as medicine.Nature and flavor are sweet, salty, can be promoting blood circulation and removing blood stasis.Alleviate spasm, pain relieving is dispelled rheumatism, and controls acute pyogenic infection of the pharynx, numbness in hands and feet, pernio, burn.Lignanoid (lignan) claim lignan again, and they are present in many kind of plant, and the position, place is different, even in the secretory product of plant, human and animal's urine discovery is arranged also; This compounds has wide biological activity, as: contained podophyllotoxin (podophyllotoxin) and the antineoplastic activity of derivative tool thereof in the multiple value things such as Berberidaceae Podophyllum emodi var chinense, schizandrin (schizandrin) can reduce gpt (GPT) and be used for the treatment of hepatitis, magnolol (maganolol) tool myorelaxant effects, Arctiin (arctiin) are effective etc. to common cold due to wind-heat.Because plant lignanoid constituent structure type is many, the stereochemistry complexity has multiple biological activity, and is very active to the research in this field both at home and abroad.The present invention separates the neolignan compounds that has obtained a kind of biologically active from Shortpetal Daphne, for the comprehensive utilization of Shortpetal Daphne provide newly by way of.
Summary of the invention
The object of the present invention is to provide a kind of new Lignanoids compounds.
Another object of the present invention provides a kind of method of extracting described compound from Shortpetal Daphne.
Further aim of the present invention provides the application of described compound in anti-oxidant and anti-HIV-1.
Purpose of the present invention is achieved by following technical proposals.
*Except as otherwise noted, the percentage ratio that is adopted among the present invention is mass percent.
A. the present invention isolates a kind of new Lignanoids compounds from Shortpetal Daphne, and this compound has significant anti-oxidant and anti-HIV-1 activity.
New Lignanoids compounds provided by the invention has following structural formula:
The called after of this compound: the A of winter daphne lignanoid (daphnelignan A).
B. the invention provides a kind of preparation method of described compound, this method adopts following steps:
1. the Shortpetal Daphne complete stool is pulverized back the branch with methyl alcohol and uses supersound extraction 3 times; 2. united extraction liquid, concentrating under reduced pressure extracting solution leave standstill back filtering throw out to small volume, and extracting solution is condensed into medicinal extract; 3. medicinal extract just divides with silica gel column chromatography, adopts high performance liquid phase half preparative chromatography further to separate then, promptly obtains this required new compound.
C. the present invention has carried out anti-oxidant to described new compound and the active detection of anti-HIV-1, and compound exhibits goes out good antioxidant activity and anti-HIV-1 effect, can be new compound or lead compound that medicine industry provides pharmaceutical use.
Description of drawings
Fig. 1 be The compounds of this invention nucleus magnetic resonance charcoal spectrum (
13C NMR);
Fig. 2 be The compounds of this invention proton nmr spectra (
1H NMR);
Fig. 3 is the high resolution mass spectrum (HRESIMS) of The compounds of this invention;
Fig. 4 is the structure contrast of The compounds of this invention and known compound Wikstromol;
Fig. 5 is that the main HMBC of The compounds of this invention is relevant.
Embodiment
In order to make purpose of the present invention, technical scheme and advantage clearer,, the present invention is described in further detail below in conjunction with drawings and Examples.Should be appreciated that specific embodiment described herein is only in order to explaining the present invention, and be not used in qualification the present invention.
Embodiment 1
---the preparation of compound
The Shortpetal Daphne sample is adopted in the Yunnan Lijing.2.0kg is crushed to 60 orders with the sampling of Shortpetal Daphne sample, and with methyl alcohol supersound extraction 3 times, extracting solution merges, and filters, and the concentrating under reduced pressure extracting solution leaves standstill back filtering throw out to small volume, is condensed into medicinal extract then; Get medicinal extract 46.5g.Medicinal extract is mixed sample with the thick silica gel of 80g (80-100 order) after with an amount of acetic acid ethyl dissolution, 0.5kg silica gel (160-200 order) dress post carry out silica gel column chromatography, sherwood oil: acetone (1: 0 → 0: 1) gradient elution, the TLC monitoring merges identical part, obtain 8 part (pure sherwood oils, sherwood oil-acetone 10: 1, sherwood oil-acetone 5: 1, sherwood oil-acetone 2: 1, sherwood oil-acetone 1: 1, sherwood oil-acetone 1: 2, sherwood oil-acetone 1: 5, pure acetone), wherein sherwood oil-acetone (2: 1) wash-out part 5.22g separates with prompt logical sequence 1,100 half preparative high-performance liquid chromatographics of peace, methyl alcohol with 65% is moving phase, ZorbaxSB-C
18(9.4 * 250mm, 5 μ m) semipreparative column is a stationary phase, it is 280nm that UV-detector detects wavelength, each sample introduction 50 μ L, the chromatographic peak of collection 21.2min, the back evaporate to dryness repeatedly adds up, use pure dissolve with methanol once more, be moving phase with pure methyl alcohol again, separate once more, can get described new compound with Sephadex LH-20 gel filtration chromatography.
Embodiment 2
---the evaluation of compound
The compounds of this invention is white amorphous powder; UV spectrum (solvent is a methyl alcohol), λ
Max(log ε): 276 (3.67), 210 (5.57) nm; Infrared spectra (pressing potassium bromide troche) 3427,2946,2857,1742,1654,1638,1535,1472,1447,1372,1219,1153,1092,1027,874cm
-1Optically-active (solvent is a methyl alcohol) [α]
D 25.3+ 18.4 (c 0.21).
Table-1 compound
1H and
13C NMR data (Pyridine-d
5)
HRESIMS shows The compounds of this invention quasi-molecular ion peak m/z 453.1521[M+Na]
+(calculated value is 453.1525), in conjunction with
1H and
13C NMR spectrum (figure-1 and figure-2, attribution data sees Table-1) provides its molecular formula C
23H
26O
8, degree of unsaturation is 11.
1H and
13C NMR spectrum signal shows an ethanoyl (δ in the compound
C169.9,21.2), 2 phenyl ring (comprise 6 two key methine carbon (δ
C112.9,113.5,114.9,116.6,121.4,123.1), 2 methylene radical (δ
C31.9,40.1), the methylene radical (δ of 1 oxidation
C71.2), 1 methyne (δ
C44.2), the season charcoal (δ of 1 oxidation
C78.0), 1 ester carbonyl group (δ
C179.5), 3 methoxyl group (δ
C55.9,56.0,56.1).The compounds of this invention
13C-NMR data and known compound Wikstromol (figure-4) comparison shows that both are closely similar, difference only is many one group of ethanoyl signals in this patent compound, illustrate with known compound Wikstromol and compare, ethanoyl replacement that The compounds of this invention is many, confirm that through HMBC relevant (figure-5) ethanoyl is substituted in the C-8 position in the The compounds of this invention, and C-3, C-3 ', C-4 ' position is substituted by methoxyl group, and it is hydroxyl that C-4 replaces the position; And a nearly step is through the relevant three-dimensional arrangement that has confirmed The compounds of this invention of ROESY.So far, the structural formula of compound finally is determined, the called after winter daphne A of lignanoid (daphnelignan A).
---compound with oxidation resistance is active to be detected
Anti-oxidant activity is represented with the size of removing DPPH free radical ability; With 50 μ g/mL is primary dcreening operation concentration, measures the activity that it removes fat free love base DPPH.Get costar 96 orifice plates, add freshly prepared DPPH methanol solution (6.5 * 10
5Mol/L) 190 μ L/ holes add testing sample 10 μ L/ holes, and blank well adds 10 μ L physiological saline, abundant mixing, left standstill 30 minutes with lucifuge under the room temperature behind the shrouding film shrouding, measure each hole absorbance on determinator on the UV2401 spectrophotometer, the mensuration wavelength is 517nm; Sample is calculated as follows fat free love base DPPH clearance rate:
DPPH clearance rate (%)=(A
Blank-A
Sample)/A
Blank* 100%
A
Blank: blank group absorbance; A
Sample: add the sample sets absorbance.
Sample detects for parallel 5 times, and calculating half removing concentration IC50 measurement result is 10.16 μ g/L, shows that compound has good antioxidant activity.
---the HIV (human immunodeficiency virus)-resistant activity of The compounds of this invention detects
1.HIV-1 infectious titration is pressed Johnson ﹠amp; The described method improvement of Byington carries out titration; Press Reed﹠amp; The Muench method is calculated the TCID of virus
50(50%Tissue Culture InfectionDose).
2. sample detects the cytotoxicity of C8166 host cell
4 * 10
5/ ml C8166 cell suspension 100ul mixes with testing compound solution, establishes three repeating holes.The control wells that does not contain compound, 37 ℃ of temperature, 5%CO are set simultaneously
2Cultivated three days, and adopted the MTT colorimetry to detect cytotoxicity.ELx800 ELISA instrument is measured the OD value, and the mensuration wavelength is 595nm, and reference wavelength is 630nm.Calculate CC
50Value (50%CytotoxicConcentration), the compound concentration during promptly to 50% normal T lymphocyte series C8166 toxigenicity.
3. sample is to HIV-1
IIIBInduce the inhibition test of C8166 cytopathy (CPE)
With 8 * 10
5/ mL C8166 cell 50 μ L/ holes are inoculated on the 96 porocyte culture plates that contain 100 μ L/ hole doubling dilution compounds, add the HIV-1 of 50 μ L then
IIIBDilution supernatant (M.O.I.0.0016).If three repeating holes.The normal cell control wells that does not contain compound is set simultaneously.37 ℃, 5%CO
2Cultivated three days, (100 *) count plasmodial formation under the inverted microscope.EC
50(50%Effective Concentration) forms 50% o'clock compound concentration for suppressing synplasm.
4. sample is to the provide protection test of HIV cells infected
With 8 * 10
5/ ml MT
4Cell 50ul/ hole is inoculated on the 96 porocyte culture plates that contain 100 μ l/ hole doubling dilution compounds, and half hole of culture plate adds the HIV-1 of 50 μ l
IIIBDilution (M.O.I.0.006), second half hole adds 50 μ l substratum.2 repeating holes of each concentration gradient are provided with control wells and the blank hole that does not contain compound, 37 ℃, 5%CO simultaneously
2Cultivate, 100 μ l fresh cultures were added in every hole in the 3rd day, adopted the MTT colorimetry to detect cell survival rate in the 5th day or the 6th day.ELx800 ELISA instrument is measured the OD value, and the mensuration wavelength is 595nm, and reference wavelength is 630nm.Calculate compound to Normocellular toxicity with to HIV-1 with formula
IIIBThe provide protection of cells infected.
5. calculation formula
Draw dose response curve according to experimental result, press Reed﹠amp; The Muench method calculates the 50% effective concentration (EC that compound suppresses virus
50), 50% cell growth inhibiting concentration (CC
50) and the active therapeutic index TI value of anti-HIV-1 (Therapeutic index) be: TI=CC
50/ EC
50
Cell growth survival rate (%)=experimental port OD value/control wells OD value * 100
Inhibitory rate of cell growth (%)=(1-experimental port OD value/control wells OD value) * 100
The cytopathogenic inhibiting rate of HIV-1 (%)=(1-experimental port synplasm number/control wells synplasm number) * 100
The protection ratio of cells infected (%)=
(experimental port OD value-positive control hole OD value)/(negative control hole OD value-positive control hole OD value) * 100
6. experimental result
Experimental result clearly illustrates that this compound exhibits goes out certain anti-HIV-1 activity, and its therapeutic index is 47.5, and having disclosed compound of the present invention has good prospects for application in the medicine of preparation AIDS resisting.
*The above only is preferred embodiment of the present invention, not in order to restriction the present invention.All any modifications of being done within the spirit and principles in the present invention, be equal to and replace and improvement etc., all should be included within protection scope of the present invention.
Claims (4)
1. compound with following structural formula:
2. the preparation method of the described compound of claim 1, this method adopts following steps:
(1) the Shortpetal Daphne sample is pulverized back the branch with methyl alcohol and is used supersound extraction 3 times, united extraction liquid;
(2) the concentrating under reduced pressure extracting solution leaves standstill back filtering throw out and is condensed into medicinal extract to small volume.
(3) medicinal extract just divides with silica gel column chromatography, adopts high performance liquid phase half preparative chromatography further to separate then, promptly obtains required compound.
3. the described compound of claim 1 is as the application of antioxidant.
4. the described compound of claim 1 is used for anti-HIV-1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101182013A CN101787004B (en) | 2010-03-05 | 2010-03-05 | Lignanoid compound contained in Yunnan daphne herb, as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010101182013A CN101787004B (en) | 2010-03-05 | 2010-03-05 | Lignanoid compound contained in Yunnan daphne herb, as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101787004A true CN101787004A (en) | 2010-07-28 |
| CN101787004B CN101787004B (en) | 2012-05-02 |
Family
ID=42530401
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010101182013A Expired - Fee Related CN101787004B (en) | 2010-03-05 | 2010-03-05 | Lignanoid compound contained in Yunnan daphne herb, as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101787004B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101973883A (en) * | 2010-09-26 | 2011-02-16 | 云南烟草科学研究院 | Phenolic compound in tobaccos and preparation method and application thereof |
| CN109197914A (en) * | 2018-09-06 | 2019-01-15 | 临沂市德力康医疗康复器械有限公司 | A kind of washing lotion of the extract containing yunnan daphne herb is improving the application in pet living environment |
| CN110538243A (en) * | 2018-05-29 | 2019-12-06 | 复旦大学 | Preparation method of active total diterpene in Thymelaeaceae plant and application thereof in pharmacy |
-
2010
- 2010-03-05 CN CN2010101182013A patent/CN101787004B/en not_active Expired - Fee Related
Non-Patent Citations (5)
| Title |
|---|
| 《Nitric Oxide》 20060228 Bhushan Shashi,Singh Jaswant,Rao J.Madhusudana,et al A novel lignan composition from Cedrus deodara induces apoptosis and early nitric oxide generation in human leukemia Molt-4 and HL-60 cells 第72-87页 1-4 第14卷, 第1期 2 * |
| 《中国中药杂志》 20090630 赵琪，刘娟等 短柄小连翘中的木脂素类化学成分研究 第1374页"2 提取分离"部分 2 第34卷, 第11期 2 * |
| 《化学进展》 20030731 杨毅，张成路，王喆，潘鑫复 木脂素抗艾滋病病毒研究 参见对比文件1第328页右栏第3-5段 1、3、4 第15卷, 第4期 2 * |
| 《现代农业科技》 20080930 郭铁英、李名扬 木脂素类化合物的研究进展 第199-201页 1-4 , 第9期 2 * |
| 《药学学报》 20031231 冯卫生、郑晓珂、王彦志、毕跃峰 马尾松松针中木脂素类成分的分离与鉴定 第927-930页 1-4 第38卷, 第12期 2 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101973883A (en) * | 2010-09-26 | 2011-02-16 | 云南烟草科学研究院 | Phenolic compound in tobaccos and preparation method and application thereof |
| CN101973883B (en) * | 2010-09-26 | 2013-05-29 | 云南烟草科学研究院 | Phenolic compound in tobaccos and preparation method and application thereof |
| CN110538243A (en) * | 2018-05-29 | 2019-12-06 | 复旦大学 | Preparation method of active total diterpene in Thymelaeaceae plant and application thereof in pharmacy |
| CN109197914A (en) * | 2018-09-06 | 2019-01-15 | 临沂市德力康医疗康复器械有限公司 | A kind of washing lotion of the extract containing yunnan daphne herb is improving the application in pet living environment |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101787004B (en) | 2012-05-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103524472B (en) | Phenolic compound, and preparation method and application thereof | |
| CN103304530A (en) | Coumarin compound and preparation method and application thereof | |
| CN105348192A (en) | Antiviral-activity isoquinoline alkaloid compound in Cassia alata L. and preparation method of antiviral-activity isoquinoline alkaloid compound | |
| CN103554077B (en) | Chromone compound as well as preparation method and application thereof | |
| CN101787004B (en) | Lignanoid compound contained in Yunnan daphne herb, as well as preparation method and application thereof | |
| CN102775375B (en) | Chromone compound, preparation method and application of chromone compound, anti-aids pharmaceutical composition prepared from chromone compound and preparation of anti-aids pharmaceutical composition | |
| CN110357894B (en) | Tricyclic alkaloid compound and preparation method and application thereof | |
| CN101555206B (en) | Angustifolia lignans, preparing method and application thereof | |
| CN104829580A (en) | Isoflavone compound contained in tobacco and preparation method and application thereof | |
| CN103896755B (en) | A kind of chalcone compounds preparation method | |
| CN113200911B (en) | Quinoline alkaloid compound and preparation method and application thereof | |
| CN102320947B (en) | Polyphenol compound contained in tobacco, preparation method and application thereof | |
| CN101696202B (en) | Phenylpropanoid compound contained in rhodiola, preparation method thereof and application thereof | |
| CN101648929B (en) | Lignanoid-like compound contained in cabo and preparation method and application thereof | |
| CN101648857B (en) | Dihydrochalcone-like compound contained in tobacco and preparation method and application thereof | |
| CN103113196B (en) | Glechoma longituba phenol, and preparation method and application thereof | |
| CN101429107B (en) | Lignanoid compounds, producing method, identification method and uses thereof | |
| CN103012099B (en) | Fluorenone compounds and preparation method and application thereof | |
| CN102786529B (en) | Pterocarpan compound and preparation process and application thereof | |
| CN110372707B (en) | Benzazepine alkaloid compound and preparation method and application thereof | |
| CN111518070B (en) | Rotavirus-resistant compound in cassia wingnut, preparation method and application thereof | |
| CN103922913B (en) | A kind of Chalcone Compounds and its preparation method and application | |
| CN107445934A (en) | A kind of flavone compound and its preparation method and application | |
| CN103664862B (en) | Polyphenolic compound and its preparation method and application in a kind of Turkish tobaccos | |
| CN101597229B (en) | Acetoxy-benzo-ketones compound, preparation method and application thereof |
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 | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120502 Termination date: 20130305 |