CN109721636A - A kind of novel triterpene saponin compound acanthopanax gracilistylus glycosides S and preparation method thereof - Google Patents
A kind of novel triterpene saponin compound acanthopanax gracilistylus glycosides S and preparation method thereof Download PDFInfo
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- CN109721636A CN109721636A CN201811349260.4A CN201811349260A CN109721636A CN 109721636 A CN109721636 A CN 109721636A CN 201811349260 A CN201811349260 A CN 201811349260A CN 109721636 A CN109721636 A CN 109721636A
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Abstract
The present invention provides a kind of novel triterpene saponin compound and preparation method thereof, and specifically, the molecular formula of noval chemical compound of the present invention is C48H76O20Entitled 1 β of chemistry of compound, 3 alpha-dihydroxys-lupin -20 (29)-alkene -23,28- diacid -28-O- [α-L- rhamnose-(1 → 4)-β-D-Glucose-(1 → 6)-β D-Glucose]-glycosides, structural formula are determined by wave spectrum analysis method.It is specific the preparation method comprises the following steps: using acanthopanax gracilistylus leaf as raw material, it is extracted after crushing with organic solvent, it is water-dispersible after filtrate concentration, after low polar solvent extracts depigmentaton, aliphatic ester, extracting n-butyl alcohol are used respectively, it takes n-butyl alcohol extract to be separated on high-speed counter-current chromatograph, after on-line ultraviolet detects, is isolated and purified to obtain noval chemical compound shown in formula 1 with preparation liquid phase.
Description
Technical field
The present invention relates to field of medicaments, in particular to a kind of novel triterpene saponin compound acanthopanax gracilistylus glycosides S and its preparation side
Method.
Background technique
Acanthopanax gracilistylus (Acanthopanax gracilistylus W.W.Smith) is studies on plants of Acanthopanax Miq. in Araliaceae, root
Skin is its main medicinal effects, has the effect of strengthening muscles and bones, dispelling wind and eliminating dampness, inducing diuresis for removing edema, tonify the liver and kidney, inducing diuresis for removing edema.
By the research of its root skin chemical component of recent decades scientific worker, contain lilac as medicinal acanthopanax gracilistylus root skin
The ingredients such as glycosides, cupreol, acanthoic acid, Syringin 1, shell olefin(e) acid, daucosterol, various organic acids, pigment, volatile oil.
In recent years, domestic and international positive the omnibearing exploration platymiscium, including stem, leaf and fruit etc., pharmacological experiments show acanthopanax gracilistylus
Lupinane type triterpene compound abundant can inhibit lipopolysaccharide-induced macrophage RAW264.7, inflammatory factor in leaf
The secretion of TNF-α, IL-1 β, HMGB1, triterpene compound have exploitation well in fields such as anti-inflammatory drug, liver disease drugs
Prospect, the system research by nearly more than ten years to acanthopanax gracilistylus leaf, for the first time isolated natural new chemical combination from acanthopanax gracilistylus leaf
Object acanthopanax gracilistylus glycosides S.
Summary of the invention
The purpose of the present invention is to provide a kind of novel triterpene saponin compound acanthopanax gracilistylus glycosides S and preparation method thereof.
A kind of noval chemical compound isolated from acanthopanax gracilistylus leaf of the invention, the chemical name of noval chemical compound are as follows: 1 β, 3 α-
Dihydroxy-lupin -20 (29)-alkene -23,28- diacid -28-O- [α-L- rhamnose-(1 → 4)-β-D-Glucose-(1 → 6) -
β-D-Glucose]-glycosides, it is named as acanthopanax gracilistylus glycosides S, chemical structural formula are as follows:
The preparation method for the novel triterpene saponin compound isolated from acanthopanax gracilistylus leaf, which is characterized in that including following
Step:
(1) acanthopanax gracilistylus leaf is weighed, the solvent extraction of 6-15 times of volume is added after crushing, it is water-dispersible after filtrate concentration,
After low polar solvent extracts depigmentaton, aliphatic ester, extracting n-butyl alcohol are used respectively, obtains aliphatic ester and n-butyl alcohol extract；
(2) take n-butyl alcohol extract with the aliphatic ester of volume ratio 0.8-5: 0.1-0.9: 0.5-2.0: 1-6: fatty alcohol:
Methanol: water is separated on high-speed counter-current chromatograph as two phase solvent system, which is uniformly mixed after standing,
Phase up and down, ultrasonic 15-40min are separated, the upper phase of solvent system is stationary phase, and lower phase is mobile phase, with the stream of 12-40ml/min
Upper phase is pumped into pipeline by speed, and after phase to be fixed is full of entire pipeline, termination of pumping starts host, then with the flow velocity of 1.5-3ml/min
It is pumped into mobile phase, when balancing each other up and down, the volume of record stationary phase outflow is calculated the retention of stationary phase, takes step
(1) n-butyl alcohol extract prepared, is dissolved in stationary phase, sample introduction, according to ultraviolet detection spectrogram, collects component, is concentrated, dry
Obtain crude product.
(3) above-mentioned dry crude product is separated with preparative high-performance liquid chromatographic, chromatographic column is to prepare column CST C18
(300*30mm, 10 μm), mobile phase are acetonitrile-water (23: 77, v/v) isocratic elution, Detection wavelength 210nm, flow velocity 20ml/
Min, collects appearance solution, and concentration is drying to obtain novel triterpene saponin compound acanthopanax gracilistylus glycosides S.
The preparation method for the novel triterpene saponin compound isolated from acanthopanax gracilistylus leaf, which is characterized in that in step (1)
Extraction solvent be one of methanol, ethyl alcohol, water or a variety of.
The preparation method for the novel triterpene saponin compound isolated from acanthopanax gracilistylus leaf, which is characterized in that step (2)
In, two-phase solvent system uses four kinds of ethyl acetate, n-butanol, first alcohol and water volume components ratios for 3: 0.3: 0.8: 4, before use
Phase up and down is separated, ultrasonic 40min, using the upper phase of solvent system as stationary phase, lower phase is mobile phase, will with the flow velocity of 25ml/min
Upper phase is pumped into pipeline, and full of after entire pipeline, termination of pumping starts host, revolving speed is transferred to 900r/min band stationary phase, then with
The flow pump of 2ml/min enters mobile phase, sample volume 15ml, and ultraviolet detection wavelength is 210nm, according to ultraviolet detection spectrogram, receives
Collect component, concentration, dry crude product.
Detailed description of the invention
Fig. 1 acanthopanax gracilistylus glycosides S's1H-NMR schemes (400MHz, CD3OD)
Fig. 2 acanthopanax gracilistylus glycosides S's13C-NMR schemes (100MHz, CD3OD)
Fig. 3 acanthopanax gracilistylus glycosides S's is main1H-1HCOSY (runic) is related to HMBC (arrow)
The main NOESY of Fig. 4 acanthopanax gracilistylus glycosides S is related
Specific embodiment
Noval chemical compound of the present invention is extracted after crushing with organic solvent, filtrate is dense using acanthopanax gracilistylus leaf as raw material
It is water-dispersible after contracting, after low polar solvent extracts depigmentaton, aliphatic ester, extracting n-butyl alcohol are used respectively, takes n-butyl alcohol extract
It is separated on high-speed counter-current chromatograph, after on-line ultraviolet detects, isolates and purifies to obtain with preparation liquid phase.Below in conjunction with reality
Border situation, a specific embodiment of the invention are described in detail:
One, the extraction and separation of compound
(1) acanthopanax gracilistylus leaf is weighed, the methanol that 8 times of volumes are added after crushing extracts, water-dispersible, petroleum after filtrate concentration
After ether extracts depigmentaton, ethyl acetate, extracting n-butyl alcohol are used respectively, obtains ethyl acetate and n-butyl alcohol extract；
(2) n-butyl alcohol extract is using the ethyl acetate of volume ratio 2: 0.7: 1.5: 4: n-butanol: methanol: water mixes as two
Agent system is separated on high-speed counter-current chromatograph, which is uniformly mixed after standing, and phase up and down, ultrasound are separated
20min, the upper phase of solvent system are stationary phase, and lower phase is mobile phase, upper phase are pumped into pipeline with the flow velocity of 25ml/min, to solid
Full of after entire pipeline, termination of pumping starts host, then enters mobile phase with the flow pump of 2ml/min, balance each other when up and down fixed phase
When, the retention of stationary phase is calculated in the volume of record stationary phase outflow, and the n-butyl alcohol extract for taking step (1) to prepare is molten
Solution is in stationary phase, sample introduction, detects spectrogram according to detector, collects component, is concentrated, dry.
(3) above-mentioned dry crude product is separated with preparation liquid phase, chromatographic column be prepare column CST C18 (300*30mm, 10
μm), mobile phase is acetonitrile-water (23: 77, v/v) isocratic elution, and Detection wavelength 210nm, flow velocity 20ml/min collect appearance
Solution, solution concentration, is drying to obtain effective component.
Two, compound structure is identified
White powder (MeOH), Liebermann-Burchard reaction and Molish reaction are positive, thus it is speculated that are triterpene
Compound or its saponin(e.MS show that its molecular ion peak is m/z972.5 [M]+, high resolution mass spectrum ESI anionic molecule amount is
971.4865 with molecular formula C48H75O20[M-H]-1Theoretical value (971.4857) coincide, infer its molecular formula be C48H76O20。
Nuclear magnetic resonance modal data is as shown in table 1.1H-NMR (400MHz, CD3OD) following signal: five methyl is shown in spectrum
Peak δ 0.95,0.98,1.03,1.09 and 1.70 (3H, s), a methylene peak δ 1.25 (3H, d, J=4.96Hz), two allyls
The sub- δ 4.58 (1H, brs) of matrix and 4.72 (1H, brs), two methine protons δ 3.67 (1H, m) and 3.80 (1H, m).Three
Anomeric proton peak δ 4.37 (1H, d, J=6.28Hz), 4.84 (1H, overlapped) and 5.45 (1H, d, J=6.56Hz), card
There are three sugar to be connected on the bright compound aglycon.
13C-NMR (100MHz, CD3OD) spectrum display contain 48 carbon signals, DEPT spectrum display containing 6 primary carbons, 13 it is secondary
Carbon, 21 tertiary carbons and 8 quaternary carbons.Wherein 2 ethylene linkage carbon δ 151.77 and δ 110.41 are that the outer end of lupin alkane type triterpenoid ring is double
The characteristic signal of key: δ 176.4 and 182.60 is the carbon signal peak of carbonyl: it is composed in conjunction with DEPT, δ 13.17,15.10,17.14,
17.84,18.08,19.49 be 6 methyl carbon signal peaks；There are a series of typical sugared hydroxyl carbon signal peaks, root between δ 60-105
According to three sugared anomeric protons, it is inferred on aglycon replace containing two hydroxyls.
1D-NMR spectrum is combined with 2D-NMR spectrum, the pass being connected directly between carbon and hydrogen has been determined by hsqc spectrum
System, by1H-1The adjacent even correlativity of H COSY spectrum display has determined that long-range relationship between carbon and hydrogen, NOESY spectrum determine by HMBC spectrum
NOE relationship between carbon and hydrogen.Mainly1H-1For H COSY and HMBC correlation as shown in table 1 and Fig. 3, NOESY is as shown in Figure 4.
NOSEY spectrum has determined NOE relationship between hydrogen and hydrogen, also thereby determines that on aglycon containing two hydroxyls, is located at 1 and 3.With
3 replace relevant periphery C chemical shift C-5 (δ 46.11), the chemical potential phase shift of C-24 (δ 18.08) and 3- β related compound
Than C-5 (δ 52.4), the significantly different (document: Adam G., Lischewski M., Phiet H.V., Preiss of C-24 (δ 11.3)
A., Schmidt J., Sung T.V., Phytochemistry, 21,1385-1387 (1982)), tentatively it is judged as that α type takes
In generation, another aspect NOESY, which composes upper 3 hydrogen and 5 hydrogen, does not have NOE effect, has NOE effect with 24 methyl, it was demonstrated that 3 hydrogen bond positions
E key illustrates that 3-OH substitutions should be α type and replace (a key), and-the COOH on the position C-4 is also that α type replaces；1 hydrogen and 2,5,25
NOE effect occurs for hydrogen, is inferred to 1-OH and is substituted by the substitution of β type.In HMBC spectrum, δ 176.4 (C-28) and δ 1.68 (H-18), δ
1.48 is related to 1.94 (H-22)；δ 182.60 (C-23) and δ 1.09 (H-24), δ 1.87 (H-5) is related, thus judges two carbonyls
Base is connected with C17 and C4 respectively.19 H chemical shift of proton are that 3.0 and 28 carbon chemical shifts are 176.40, and C-28 exists
It is related to sugared anomeric proton δ 5.45 (glcH-1) in HMBC spectrum, show that 28 replace for glycosidic bond.HMBC is composed while being given
Three sugared specific orders of connection.
1H-NMR (400MHz, CD3OD) and13C-NMR (100MHz, CD3OD) all 1D-NMR and 2D-NMR data of data are all
Confirmation compound 1 is 1 β of lupinane type triterpenoid saponin, 3 alpha-dihydroxys-lupin -20 (29)-alkene -23,28- diacid -28-O-
[α-L- rhamnose-(1 → 4)-β-D-Glucose-(1 → 6)-β-D-Glucose] glycosides.
1 compound 1 of table1H-NMR and13C-NMR data (CD3OD, δ ppm)
The basic principles, main features and the advantages of the invention have been shown and described in above embodiments.The industry
Technical staff it should be appreciated that the present invention is not limited to the above embodiments, what is described in the above embodiment and the description is only
Illustrate the principle of the present invention, rather than limits the scope of the invention in any way, without departing from the scope of the invention,
Various changes and improvements may be made to the invention, these changes and improvements are both fallen in claimed range.The present invention claims guarantors
Shield range is defined by the appending claims and its equivalent thereof.
Claims (4)
1. a kind of from the isolated novel triterpene saponin compound of acanthopanax gracilistylus leaf, which is characterized in that the structural formula of the compound
As shown in Equation 1, chemical name are as follows: 1 β, 3 alpha-dihydroxys-lupin -20 (29)-alkene -23,28- diacid -28-O- [α-L- rhamnose -
(1 → 4)-β-D-Glucose-(1 → 6)-β-D-Glucose]-glycosides, it is named as acanthopanax gracilistylus glycosides S.
2. the preparation method of the novel triterpene saponin compound according to claim 1 isolated from acanthopanax gracilistylus leaf,
It is characterized in that, comprising the following steps:
(1) acanthopanax gracilistylus leaf is weighed, the solvent extraction of 6-15 times of volume, water-dispersible, low pole after filtrate concentration are added after crushing
Property solvent extraction depigmentaton after, respectively use aliphatic ester, extracting n-butyl alcohol, obtain aliphatic ester and n-butyl alcohol extract；
(2) take n-butyl alcohol extract with the aliphatic ester of volume ratio 0.8-5: 0.1-0.9: 0.5-2.0: 1-6: fatty alcohol: methanol:
Water is separated on high-speed counter-current chromatograph as two phase solvent system, which is uniformly mixed after standing, is separated
Phase up and down, ultrasonic 15-40min, the upper phase of solvent system are stationary phase, and lower phase is mobile phase, will with the flow velocity of 12-40ml/min
Upper phase is pumped into pipeline, and after phase to be fixed is full of entire pipeline, termination of pumping is started host, is then pumped into the flow velocity of 1.5-3ml/min
Mobile phase, when balancing each other up and down, the volume of record stationary phase outflow is calculated the retention of stationary phase, step (1) is taken to make
Standby n-butyl alcohol extract, is dissolved in stationary phase, sample introduction, according to ultraviolet detection spectrogram, collects component, concentration is slightly dry
Product.
(3) above-mentioned dry crude product is separated with preparative high-performance liquid chromatographic, chromatographic column is to prepare column CST C18 (300*
30mm, 10 μm), mobile phase is acetonitrile-water (23: 77, v/v) isocratic elution, and Detection wavelength 210nm, flow velocity 20ml/min are received
Collect appearance solution, concentration is drying to obtain novel triterpene saponin compound acanthopanax gracilistylus glycosides S.
3. the preparation method of the novel triterpene saponin compound according to claim 2 isolated from acanthopanax gracilistylus leaf,
It is characterized in that, the Extraction solvent in step (1) is one of methanol, ethyl alcohol, water or a variety of.
4. the preparation method of the novel triterpene saponin compound according to claim 2 isolated from acanthopanax gracilistylus leaf,
It is characterized in that, in step (2), two-phase solvent system uses four kinds of ethyl acetate, n-butanol, first alcohol and water volume components ratios for 3:
0.3: 0.8: 4, phase up and down is separated before use, ultrasonic 40min, using the upper phase of solvent system as stationary phase, lower phase is mobile phase, with
Upper phase is pumped into pipeline by the flow velocity of 25ml/min, and after phase to be fixed is full of entire pipeline, termination of pumping starts host, revolving speed is transferred to
Then 900r/min enters mobile phase, sample volume 15ml with the flow pump of 2ml/min, ultraviolet detection wavelength is 210nm, according to
Ultraviolet detection spectrogram collects component, concentration, dry crude product.
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| CN116903576A (en) * | 2023-09-13 | 2023-10-20 | 江西中医药大学 | Triterpene compound in syringa oblata, and preparation method and application thereof |
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| CN116903576A (en) * | 2023-09-13 | 2023-10-20 | 江西中医药大学 | Triterpene compound in syringa oblata, and preparation method and application thereof |
| CN116903576B (en) * | 2023-09-13 | 2023-11-24 | 江西中医药大学 | Triterpene compound in syringa oblata, and preparation method and application thereof |
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Application publication date: 20190507 |