CN102826982A - Monoterpene compounds prepared from Senecio cannabifolius Less, and extraction method and application thereof - Google Patents
Monoterpene compounds prepared from Senecio cannabifolius Less, and extraction method and application thereof Download PDFInfo
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Abstract
The invention provides monoterpene compounds prepared from Senecio cannabifolius Less. Specifically speaking, the monoterpene compounds are compound I and compound II. The monoterpene compounds are prepared through the following steps: crushing of Senecio cannabifolius Less; heating extraction and condensation; separation with a positive phase silicagel column and elution; condensation and drying; and separation by using prepared liquid chromatogram. According to the invention, the monoterpene compounds have good anti-inflammatory activity and can be used in preparation of anti-inflammatory drugs; the general formula of the monoterpene compounds is described in the specification.
Description
Technical field
The invention belongs to the Chinese drug field, extraction, separation and the preparation technology field of pharmaceutically active ingredient relate in particular to extraction separation has anti-inflammatory activity in the Chinese medicine Herba Senecionis cannabifolii compound and application thereof in being specifically related to.
Background technology
Herba Senecionis cannabifolii is the dry aerial parts of composite family Senecio per nnial herb Senecio cannabifolius (Senecio cannabifolius Less.), has that heat-clearing is eliminated the phlegm, the effect of cough-relieving.Modern pharmacology research shows, that Herba Senecionis cannabifolii has is antibiotic, anti-inflammatory and effect such as antiviral (Yang Xiudong etc., Changchun University of Traditional Chinese Medicine's journal, 2006,22,70-71.); The preparation of at present having been developed has the peaceful particle of lung (Herba Senecionis cannabifolii particle), the peaceful oral liquid of lung etc.Be usually used in treating the chronic bronchitis cough clinically.Have liposoluble ingredient in the research report Herba Senecionis cannabifolii have anti-microbial activity (Wu Bin etc. herbal medicine, 2005,36 (10): 1447-1450; Wu Bin etc. Chinese pharmaceutical chemistry magazine, 2005,15 (3): 178～179; Wu Bin etc., research and development of natural products, 2005,17 (4): 440-443.), but the chemical ingredients that has anti-inflammatory activity in the Herba Senecionis cannabifolii it be unclear that.
Summary of the invention
The purpose of this invention is to provide a kind of monoterpenes compound that from Herba Senecionis cannabifolii, extracts, the general formula of described monoterpene is following:
Wherein:
Chemical compounds I: R
1+ R
2=O, R
3=OH, R
4=H, △
2,3,
Compound ii: R
1=OH, R
2=H, R
3=R
4=O.
The structural formula of chemical compounds I and compound ii is following:
The chemical compounds I compound ii
Specifically have chemical compounds I (E, 4R)-4-hydroxyl-4,5; 5-trimethylammonium-3-(3-oxo butylene) tetrahydrobenzene-2-ketone [(E, 4S)-4-hydroxy-4,5; 5-trimethyl-3-(3-oxobut-1-enyl) cyclo-hex-2-enone] and compound ii (E)-4-((1S, 3R, 4R)-1-hydroxyl-4; 5,5-trimethylammonium-7-oxo bridged ring [4.1.0] heptene)-1-butylene-3-ketone [(E, 3S)-4-(3-hydroxy-5; 5,6-trimethyl-7-oxabicyclo [4.1.0] heptan-1-yl) but-3-en-2-one].
Another object of the present invention is the preparation method of the monoterpenes compound that from Herba Senecionis cannabifolii, extracts, realizes through following steps: extract solvent with adding after the Herba Senecionis cannabifolii pulverizing medicinal materials, heating and extracting gets extracting solution; Extracting solution is condensed into medicinal extract, and itself and silica gel are mixed appearance, it is separated with normal phase silicagel column; With sherwood oil and ETHYLE ACETATE as eluting solvent; Get elutriant, elutriant obtains component behind concentrate drying, separates obtaining chemical compounds I and compound ii again through preparative liquid chromatography.
Preferred chemical compounds I and compound ii preparation method comprise the following steps: after the Herba Senecionis cannabifolii pulverizing medicinal materials, add 3-8 times of water gaging or 10% ~ 90% ethanol-water solution; Reflux 1-2 hour, extract united extraction liquid 1-3 time; Extracting solution is condensed into medicinal extract, and itself and silica gel are mixed appearance, it is separated with normal phase silicagel column; At first use volume ratio as the sherwood oil (60-90 ℃) of 5:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant E, abandon it.Use again volume ratio as the sherwood oil (60-90 ℃) of 3:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant F.Elutriant F concentrate drying obtains chemical compounds I and two monoterpenes compounds of II after preparative liquid chromatography separates.The separation condition of preparative hplc: chromatographic column is preparative column Agilent Zorbax SB-C
18Post 250 * 21.2mm, 7 μ m, moving phase is water and methyl alcohol, the gradient elution program is following: 0min, 10% methanol-water; 60min, 50% methanol-water; Flow velocity: 10ml/min, column temperature are room temperature, detect wavelength: 254nm.Collect the chromatographic peak of 31.0min, decompression and solvent recovery gets chemical compounds I.Collect the chromatographic peak of 34.5min, decompression and solvent recovery gets compound ii.
A further object of the present invention is the application of monoterpenes compound in the preparation anti-inflammatory drug of from Herba Senecionis cannabifolii, extracting.
Chemical compounds I of the present invention and compound ii can be used as activeconstituents, add drug excipient or the carrier accepted on the pharmaceutics, process preparation according to the method for putting down in writing on the pharmaceutics.The dosage form of said medicine comprises granule, tablet, electuary, powder, oral liquid, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, sucks agent, granule, pill, paste, sublimed preparation, sprays, pill, disintegrating agent, oral cavity disintegration tablet, micropill etc.
Beneficial effect of the present invention is: (1) the present invention discloses two kinds of All new compounds in the Chinese medicine Herba Senecionis cannabifolii first and the extraction preparation method of these two kinds of new compounds is provided.(2) the invention provides the anti-inflammatory activity evaluation result of two kinds of new compounds on LPS stimulation RAW246.7 cell strain in the Herba Senecionis cannabifolii, find that it all has anti-inflammatory activity preferably, can be developed as anti-inflammatory drug.
Embodiment
The present invention combines embodiment further explain flesh and blood of the present invention, and this embodiment only is used to the present invention is described and is not limitation of the present invention.
The extraction of chemical compounds I and compound ii preparation in embodiment one Herba Senecionis cannabifolii
After the Herba Senecionis cannabifolii pulverizing medicinal materials, add 8 times of water gagings, reflux 2 hours is extracted 3 times; United extraction liquid is condensed into medicinal extract with extracting solution, and itself and silica gel are mixed appearance; With normal phase silicagel column it is separated, at first use volume ratio as the sherwood oil (60-90 ℃) of 5:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant E; Abandon it, use again volume ratio as the sherwood oil (60-90 ℃) of 3:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant F.Elutriant F concentrate drying obtains two monoterpenes compounds after preparative liquid chromatography separates.The separation condition of preparative hplc: chromatographic column is preparative column AgilentZorbax SB-C
18Post 250 * 21.2mm, 7 μ m, moving phase is water and methyl alcohol, the gradient elution program is following: 0min, 10% methanol-water; 60min, 50% methanol-water; Flow velocity: 10ml/min, column temperature are room temperature, detect wavelength: 254nm.Collect the chromatographic peak of 31.0min, decompression and solvent recovery gets chemical compounds I.Collect the chromatographic peak of 34.5min, decompression and solvent recovery gets compound ii.
The extraction of chemical compounds I and compound ii preparation in embodiment two Herba Senecionis cannabifoliis
After the Herba Senecionis cannabifolii pulverizing medicinal materials, add 3 times of amount 70% ethanol-water solutions, reflux 1.5 hours is extracted 1 time; United extraction liquid is condensed into medicinal extract with extracting solution, and itself and silica gel are mixed appearance; With normal phase silicagel column it is separated, at first use volume ratio as the sherwood oil (60-90 ℃) of 5:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant E; Use again volume ratio as the sherwood oil (60-90 ℃) of 3:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant F, abandon it.Elutriant F concentrate drying obtains two monoterpenes compounds after preparative liquid chromatography separates.The separation condition of preparative hplc: chromatographic column is preparative column Agilent Zorbax SB-C
18Post 250 * 21.2mm, 7 μ m, moving phase is water and methyl alcohol, the gradient elution program is following: 0min, 10% methanol-water; 60min, 50% methanol-water; Flow velocity: 10ml/min, column temperature are room temperature, detect wavelength: 254nm.Collect the chromatographic peak of 31.0min, decompression and solvent recovery gets chemical compounds I.Collect the chromatographic peak of 34.5min, decompression and solvent recovery gets compound ii.
The extraction of chemical compounds I and compound ii preparation in embodiment three Herba Senecionis cannabifoliis
After the Herba Senecionis cannabifolii pulverizing medicinal materials, add 6 times of amount 20% ethanol-water solutions, reflux 1 hour is extracted 2 times; United extraction liquid is condensed into medicinal extract with extracting solution, and itself and silica gel are mixed appearance; With normal phase silicagel column it is separated, at first use volume ratio as the sherwood oil (60-90 ℃) of 5:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant E; Abandon it, use again volume ratio as the sherwood oil (60-90 ℃) of 3:1 and ETHYLE ACETATE as the eluting solvent wash-out, elutriant F.Elutriant F concentrate drying obtains two monoterpenes compounds after preparative liquid chromatography separates.The separation condition of preparative hplc: chromatographic column is preparative column Agilent Zorbax SB-C
18Post 250 * 21.2mm, 7 μ m, moving phase is water and methyl alcohol, the gradient elution program is following: 0min, 10% methanol-water; 60min, 50% methanol-water; Flow velocity: 10ml/min, column temperature are room temperature, detect wavelength: 254nm.Collect the chromatographic peak of 31.0min, decompression and solvent recovery gets chemical compounds I.Collect the chromatographic peak of 34.5min, decompression and solvent recovery gets compound ii.
Chemical compounds I and compound ii structure are identified in embodiment four Herba Senecionis cannabifoliis
Chemical compounds I is a red oil, [α]
20 D=+10.8 (c=0.1, MeOH); IR (KBr): υ
Max3447,1659,1188,1122,1027,987cm
-1 1H spectrum (DMSO, 500MHz) with
13The C spectrum (DMSO, 125MHz), its NMR data see the following form 1; HR-ESI-MS m/z: molecular ion peak 221.1180 [M-H]
-(C
13H
17O
3, calculated value 221.1183).
Table 1
The nuclear-magnetism C-H of chemical compounds I is long-range to be correlated with as follows:
H-2 and C-4, C-6, H-6 and C-1, C-4, H-7 and C-3, C-4 exist long-range coherent signal between H-8, H-9 and the C-6, have 4-hydroxyl-4,5,5-trimethylcyclohexyl-2-ketenes ring in this explanation chemical compounds I molecule.δ 6.28 (d, J=16.0Hz, H-2 '), 6.99 (d, J=16.0Hz, H-1 '), there is (E)-ketone crotonyl in these three hydrogen signal explanation compound molecules of 2.29 (d, H-4 ').H-1 ' and C-3, long-range coherent signal explanation (the E)-ketone crotonyl between the C-4 is connected 4-hydroxyl-4,5, on 3 carbon of 5-trimethylcyclohexyl-2-ketenes ring.
Through
1The H spectrum,
13C spectrum, two-dimensional spectrum identify obtain the chemical compounds I structure for (E, 4R)-4-hydroxyl-4,5,5-trimethylammonium-3-(3-oxo butylene) tetrahydrobenzene-2-ketone.
The The compounds of this invention II is a red oil, [α]
20 D=-4.1 (c=0.1, MeOH); IR (KBr): υ
Max3432,1676,1258,1177,1042,989cm
-1 1H spectrum (DMSO, 500MHz) with
13The C spectrum (DMSO, 125MHz) nuclear magnetic resonance data sees the following form 2; HR-ESI-MS m/z: molecular ion peak 223.1341 [M-H]
-(calculated value C
13H
19O
3, 223.1340).
Table 2
The nuclear-magnetism C-H of compound ii is long-range to be correlated with as follows:
Different with chemical compounds I is δ
C194.7 ketone group carbon and δ
C127.0, the substitute is two oxo carbon atom (δ with 162.1 olefinic carbon blackout
C62.2 with 67.4) and a δ
C40.8 carbon atom.H-2 and C-4, C-6, H-6 and C-4, H-7 and C-3 exist long-range coherent signal between H-8, H-9 and the C-6, have the hexanaphthene skeleton in this explanation compound ii molecule.δ 6.02 (d, J=15.5Hz, H-2 '), 7.09 (d, J=15.5Hz, H-1 '), there is (E)-ketone crotonyl in these three hydrogen signal explanation compound molecules of 2.24 (d, H-4 ').H-1 ' and C-4 exist long-range coherent signal to explain that (E)-ketone crotonyl is connected 3 of hexanaphthene skeleton between H-2 and the C-1 '.
Through
1The H spectrum,
13C spectrum, two-dimensional spectrum identify that obtaining the compound ii structure is (E)-4-((1S, 3R, 4R)-1-hydroxyl-4,5,5-trimethylammonium-7-oxo bridged ring [4.1.0] heptene)-1-butylene-3-ketone.
The anti-inflammatory activity evaluation of chemical compounds I and compound ii in embodiment five Herba Senecionis cannabifoliis
Set normal cell group, LPS (LPS) stimulation control group and dosing group in the anti-inflammatory activity evaluation experimental.
Use the LPS of 200ng/ml to stimulate the RAW264.7 scavenger cell, the chemical compounds I and the compound ii that add 0,1.5625,3.125,6.25,12.5,25,50 μ mol/L concentration are respectively hatched, and the normal cell group does not add LPS to stimulate.Hatch and adopt the nitrogen protoxide detection kit to detect nitric oxide production secretory volume in the cell culture fluid after 24 hours.Measuring method adopts test kit that method is provided; Be specially and get 100 μ l normal group, control group and dosing group cell culture fluid and a series of concentration Sodium Nitrite standard solution adds in 96 orifice plates; Add Griess reagent I and reagent II respectively; The vibration mixing is placed in the ELIASA, selects 550nm to measure each hole absorbance, calculates anti-inflammatory activity.
Record chemical compounds I and have significant anti-inflammatory activity, the inhibiting rate that under 50 μ mol/L concentration, suppresses LPS inductive NO nucleus formation is 77.5%, and it suppresses NO excretory IC
50Value is 30.65 μ M.
Record compound ii and have certain anti-inflammatory activity, the inhibiting rate that under 50 μ mol/L concentration, suppresses LPS inductive NO nucleus formation is 47.3%.
The preparation of embodiment six dropping pill formulations
Get chemical compounds I with anti-inflammatory activity or compound ii 0.05g and 10.5g polyoxyethylene glycol-20000 and mix, heating and melting moves in the dripping pill drip irrigation after changing material, and in ℃ whiteruss of medicine liquid droplet to 6 ~ 8, oil removing makes 400 of dripping pills.
The preparation of embodiment seven oral liquids
Get chemical compounds I or compound ii 0.1g, sucrose 200g and zero(ppm) water 1000ml with anti-inflammatory activity, after said components mixed, 100 of packing promptly got after the sterilization.
Claims (4)
1. monoterpenes compound from Herba Senecionis cannabifolii is characterized in that the general formula of described monoterpene is following:
Wherein:
Chemical compounds I: R
1+ R
2=O, R
3=OH, R
4=H, △
2,3,
Compound ii: R
1=OH, R
2=H, R
3=R
4=O.
2. the preparation method of the described a kind of monoterpenes compound from Herba Senecionis cannabifolii of claim 1 is characterized in that, realizes through following steps: after the Herba Senecionis cannabifolii pulverizing medicinal materials; Add 3-8 times of water gaging or 10% ~ 90% ethanol-water solution, reflux 1-2 hour, extract 1-3 time; United extraction liquid is condensed into medicinal extract with extracting solution, and itself and silica gel are mixed appearance; With normal phase silicagel column it is separated, at first use sherwood oil and the eluent ethyl acetate of volume ratio, get elutriant E as 5:1; Abandon it, use sherwood oil and the eluent ethyl acetate of volume ratio again, get elutriant F as 3:1; Elutriant F concentrate drying obtains chemical compounds I and compound ii, the separation condition of preparative hplc after preparative liquid chromatography separates: chromatographic column is preparative column Agilent Zorbax SB-C
18Post 250 * 21.2 mm, 7 μ m, moving phase is water and methyl alcohol, the gradient elution program is following: 0 min, 10% methanol-water; 60 min, 50% methanol-water; Flow velocity: 10ml/min, column temperature are room temperature, detect wavelength: 254 nm, and the chromatographic peak of collection 31.0min, decompression and solvent recovery gets chemical compounds I, collects the chromatographic peak of 34.5min, and decompression and solvent recovery gets compound ii.
3. a kind of application of monoterpenes compound in the preparation anti-inflammatory drug from Herba Senecionis cannabifolii according to claim 1 is characterized in that said monoterpenes compound is chemical compounds I and compound ii.
4. application according to claim 3 is characterized in that, the dosage form of said medicine is solid preparation or liquid preparation.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110143991A (en) * | 2019-06-21 | 2019-08-20 | 中国科学院新疆理化技术研究所 | Monocyclic monoterpene glucoside compound and its preparation method and application |
| CN117946059A (en) * | 2024-03-27 | 2024-04-30 | 江西中医药大学 | Monoterpene compound in herba Senecionis Scandentis of radish leaves, and preparation method and application thereof |
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|---|---|---|---|---|
| CN101396389A (en) * | 2008-10-16 | 2009-04-01 | 吉林修正药业新药开发有限公司 | Senecio cannabifolius effective constituent extraction method and preparation method of granules thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101396389A (en) * | 2008-10-16 | 2009-04-01 | 吉林修正药业新药开发有限公司 | Senecio cannabifolius effective constituent extraction method and preparation method of granules thereof |
Non-Patent Citations (1)
| Title |
|---|
| YI TAO等: "Two new compounds from Senecio cannabifolius", 《JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110143991A (en) * | 2019-06-21 | 2019-08-20 | 中国科学院新疆理化技术研究所 | Monocyclic monoterpene glucoside compound and its preparation method and application |
| CN110143991B (en) * | 2019-06-21 | 2022-06-24 | 中国科学院新疆理化技术研究所 | Monocyclic monoterpene glucoside compound and preparation method and application thereof |
| CN117946059A (en) * | 2024-03-27 | 2024-04-30 | 江西中医药大学 | Monoterpene compound in herba Senecionis Scandentis of radish leaves, and preparation method and application thereof |
| CN117946059B (en) * | 2024-03-27 | 2024-06-04 | 江西中医药大学 | Monoterpene compound in herba Senecionis Scandentis of radish leaves, and preparation method and application thereof |
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Application publication date: 20121219 |