CN106674311A - Benzofuran glycoside compounds as well as preparation method and application thereof - Google Patents
Benzofuran glycoside compounds as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN106674311A CN106674311A CN201611228845.1A CN201611228845A CN106674311A CN 106674311 A CN106674311 A CN 106674311A CN 201611228845 A CN201611228845 A CN 201611228845A CN 106674311 A CN106674311 A CN 106674311A
- Authority
- CN
- China
- Prior art keywords
- benzofuran
- sample
- methanol
- extraction
- water
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to benzofuran glycoside compounds as well as a preparation method and an application thereof. The preparation method comprises steps as follows: dried roots of heracleum dissectum are taken and subjected to reflux extraction and extraction, and an n-butyl alcohol extraction layer is obtained; a sample of the n-butyl alcohol extraction layer is loaded to a silica gel column and is subjected to gradient elution with a chloroform-methanol system as an eluent, fractions in eluent volume ratios being 10:1 and 8:1 are combined respectively, a solvent is removed, and a sample a1 and a sample b1 are obtained; the sample a1 and the sample b1 are loaded on a reverse phase silica gel chromatographic column, a methanol-water system is taken as an eluent for gradient elution, fractions with an eluent volume ratio being 100:0 are combined, the solvent is removed, and a sample a2 and a sample b2 are obtained; the sample a2 and the sample b2 are loaded on an HPLC separation column and subjected to isocratic elution with a mobile phase, and the benzofuran glycoside compounds are obtained. The compounds have good anti-inflammatory action and are expected to be developed into new anti-inflammatory drugs or used for preparing healthcare food with functions of preventing and treating inflammation.
Description
【Technical field】
The invention belongs to medicine and field of health care food, and in particular to a kind of benzofuran glycosides compound and its preparation side
Method and application.
【Background technology】
Inflammation is body for a kind of defense reaction for stimulating, and is the biological tissue with vascular system to damage factor institute
The defense reaction of generation.It is broadly divided into two kinds of acute inflammation and chronic inflammatory disease.Participating in the material of inflammatory reaction has histamine, 5- hydroxyl colors
The polypeptides such as the amine substances such as amine, bradykinin, kallidin, fibrin lyase, kallikrein protein enzyme etc..
Inflammation is closely related with various diseases such as arteriosclerosis, heart disease, apoplexy, cancer, diabetes, is to cause various diseases
The key factor of disease.Therefore, the constantly research and development of the anti-inflammatory drug with high-efficiency low-toxicity feature are particularly significant.
The plant of nature and the secondary metabolite of microorganism are always the important source of new drug discovery, because natural product
Thing has an important structure diversity and diverse biological activities, and the structure of many compounds is unpredictable in advance and is difficult to use
The method of synthesis is obtained.Although with the development of synthetic pharmacochemistry, now increasing new drug is chemical medicine, but still has phase
When the new drug of ratio is directly or indirectly to derive from natural product.Therefore, remain can not during new drug development for natural product
Or a scarce important compound treasure-house.
Xingan's Radix Angelicae Pubescentiss (Heracleum dissectum) are distributed mainly on the Northeast, are that Compositae cattle Radix Saposhnikoviae belongs to perennial
Herbaceous plant, Hubei Province language is referred to as Kan Kula, Kan Kula as the important medicine-food two-purpose of the Oroqen wild plant, spring and summer season its
The tender stem and leaf of children is used as delicious vegetable and eats by locals, and its root is used for expelling wind and removing dampness, pain relieving as traditional Hubei Province race medical material
Antidiarrheal etc..Therefore, Kan Kula is considered as jewellery by the Olunchuns, is just stewed robe meat with Kan Kula when only honored guest arrives and is received
Guest.But the research up to the present to its chemical composition is few, also parmacodynamics-less activity research is reported.
【The content of the invention】
It is an object of the invention to overcome problems of the prior art, there is provided a kind of benzofuran glycosides compound and
Its preparation method and application, can be obtained a kind of new benzofuran glycosides compound, for antiinflammatory.
In order to achieve the above object, the present invention is adopted the following technical scheme that：Its molecular formula is C23H30O13Or C22H28O13。
Further, its general structure is：
Wherein, R is-H or-CH3。
Preparation method of the present invention, comprises the following steps：
1) taking the dry root of Xingan's Radix Angelicae Pubescentiss carries out reflux, extract, several times, by extracting solution concentrating under reduced pressure, obtain total extractum or
Concentrated solution；
2) total extractum is suspended in water, obtains extractum liquid, extractum liquid or concentrated solution are Jing after ethyl acetate is extracted several times
Organic layer is isolated, remaining water layer passes through n-butanol extraction rear combining extraction liquid several times, n-butyl alcohol is removed under reduced pressure and obtains positive fourth
Alcohol extract layer；
3) n-butanol extraction layer is splined on into silicagel column, using chloroform-methanol system as eluent, by volume (100:
0)~(0:100) gradient elution is carried out, effluent is detected, be 10 by effluent volume ratio:1 stream part merges, and removes
Solvent, obtains sample a1；It is 8 by effluent volume ratio:1 stream part merges, and removes solvent, obtains sample b1；
4) sample a1 is splined on into reversed-phase silica gel chromatography post, using methanol-water system as eluent, by volume (0:
100)~(100:0) gradient elution is carried out, is 100 by effluent volume ratio:0 stream part merges, and removes solvent, obtains sample
a2；
Sample b1 is splined on into reversed-phase silica gel chromatography post, using methanol-water system as eluent, by volume (0:100)
~(100:0) gradient elution is carried out, is 70 by effluent volume ratio:30 stream part merges, and removes solvent, obtains sample b2；
5) sample a2 and sample b2 are splined on into respectively high performance liquid chromatography separation post, with mobile phase isocratic elution are carried out,
Obtain benzofuran glycosides compound.
Further, step 1) reflux, extract, in using the ethanol of methanol, water or volume fraction 10~95% as carrying
Agent is taken, the dry root of Xingan's Radix Angelicae Pubescentiss is 1kg with the mass volume ratio of extractant：(1~8) L；When extractant is methanol or volume integral
During the ethanol of number 10~95%, solvent recovery in the extracting solution for obtaining is obtained into total extractum；When extractant is water, by extracting solution
Volume concentration to 1/10 to ten/6ths, obtain concentrated solution.
Further, step 1) in extraction time be 1~6 time, every time 1~4 hour.
Further, step 2) in the volume ratio of total extractum and water be 1:1~1:5.
Further, step 2) in extractum liquid or concentrated solution be directly over ethyl acetate extraction；Or first Jing oil successively
After ether and chloroform extraction, then through ethyl acetate extraction；Every time extraction is equal-volume extraction；Every kind of solvent extracts respectively 1~6
It is secondary.
Further, step 3) in a stream part is collected per 300~800mL after gradient elution；Step 4) in gradient elution
Collect a stream part per 200~500mL afterwards；Step 3) and step 4) in effluent be to carry out TLC detections；Step 5) it is medium
The flow velocity of degree eluting is 3~6mL/min.
Further, step 5) in mobile phase be the acetic acid of methanol -0.5% water system or the acetic acid water system of acetonitrile -0.5%；
When carrying out isocratic elution to sample a2, the volume ratio of methanol, water and acetic acid is for 45 in the acetic acid water system of methanol -0.5% for adopting：
55：0.5, the volume ratio of acetonitrile, water and acetic acid is for 42 in the acetic acid water system of acetonitrile -0.5% for adopting：58：0.5, the benzene for obtaining
And furan glycosides compound molecular formula is C23H30O13；When carrying out isocratic elution to sample a2, the acetic acid water of methanol -0.5% of employing
The volume ratio of methanol, water and acetic acid is 35 in system：65：0.5, acetonitrile in the acetic acid water system of acetonitrile -0.5% of employing, water and
The volume ratio of acetic acid is 31：69：0.5, the benzofuran glycosides compound molecular formula for obtaining is C22H28O13。
The application of benzofuran glycosides compound aspect in anti-inflammatory drug and/or health product is prepared as above.
Compared with prior art, the invention has the beneficial effects as follows：
New benzofuran glycosides compound in the present invention has antiinflammatory action, tests prove that, to mice
RAW264.7 small cells have good antiinflammatory action, the compound have efficiently, low toxicity the characteristics of, be expected to develop into new anti-
Scorching medicine, or for preparing the health food with prevention and treatment inflammation.
Because the polarity of the compound in the present invention is larger, it is difficult to purification with general method and obtains, it is anti-as using
The phase chromatographic column compound cannot retain, and flow out with mobile phase, it is impossible to reach and the detached purpose of other compounds, this
Bright middle utilization hydrophilic Interaction Chromatography is isolated by benzofuran glycosides compound, and purity higher (＞ 98.5%), is to separate
The small molecule aglycon of the big polarity of purification connects the effective means of multiple saccharide compounds.
【Description of the drawings】
Fig. 1 is the compounds of this invention 11H-NMR collection of illustrative plates；
Fig. 2 is the compounds of this invention 113C-NMR collection of illustrative plates；
Fig. 3 is the DEPT collection of illustrative plates of the compounds of this invention 1；
Fig. 4 is the compounds of this invention 11H-1H COSY collection of illustrative plates；
Fig. 5 is the HMQC collection of illustrative plates of the compounds of this invention 1；
Fig. 6 is the HMBC collection of illustrative plates of the compounds of this invention 1；
Fig. 7 is the compounds of this invention 21H-NMR collection of illustrative plates；
Fig. 8 is the HMQC collection of illustrative plates of the compounds of this invention 2；
Fig. 9 is the compounds of this invention 21H-1H COSY collection of illustrative plates.
【Specific embodiment】
With reference to specific embodiment, the present invention is further illustrated, but is not limited thereto.
The structural formula of the new benzofuran glycosides compound of the present invention is as follows：
Wherein, the R in structural formula is：- H or-CH3。
The preparation method of the new benzofuran glycosides compound of the present invention, comprises the following steps：
1) Xingan's Radix Angelicae Pubescentiss dry root of certain mass (kg) is taken, is Xingan's Radix Angelicae Pubescentiss dry root 1~8 times of amount of quality with volume
Methanol, volume fraction are heating and refluxing extraction 1~6 time near respective boiling point of 10~95% ethanol or water (L), every time 1~4
Hour, when the ethanol that extractant is methanol or volume fraction 10~95%, united extraction liquid recovered under reduced pressure removes solvent, obtains
Total extractum, when extractant is water, united extraction liquid and by its volume concentration to 1/10 to ten/6ths, concentrated
Liquid；
2) total extractum is suspended in water, the volume ratio of total extractum and water is 1:1~1:5, obtain extractum liquid, extractum liquid or
Concentrated solution difference is extracted successively with isopyknic organic solvent, wherein, with petroleum ether to extractum liquid or concentration when extracting for the first time
Liquid carries out equal-volume extraction, and afterwards every time extraction is to isolate the organic layer after last extraction, by remaining water layer with having
Machine solvent equal-volume is extracted next time；Respectively extraction 1~6 time of every kind of solvent, combining extraction liquid, normal pressure or vacuum distillation are removed
Organic solvent, respectively obtains each extract layer and water layer.Organic solvent includes petroleum ether, chloroform, ethyl acetate and n-butyl alcohol etc., and
Extraction order is first to use the little solvent of polarity, then can be saved with the big organic solvent of polarity, petroleum ether and chloroform.
3) n-butanol extraction layer is taken, by using separation methods such as column chromatography purification, obtaining the benzofuran glycosides of the present invention
Class compound.
Column chromatography includes the three below stage：
First stage：N-butanol extraction layer is splined on into silicagel column, using chloroform-methanol system as eluent, by volume
Than (100:0)~(0:100) gradient elution is carried out, a stream part is collected per 300~800ml, TLC detections are carried out to effluent,
Merge identical stream part, 30 stream parts are obtained, be respectively designated as FrB1-FrB30, normal pressure or evaporated under reduced pressure solvent, take therein
FrB20 stream parts and FrB24 stream parts, i.e. effluent volume ratio respectively 10:1 and 8:1 stream part, as first time post part is crossed；
Second stage：The FrB20 stream parts that first time is crossed in post part and FrB24 stream parts, are splined on respectively reverse phase silica gel
Chromatographic column, using methanol-water system as eluent, by volume (0:100)~(100:0) gradient elution is carried out, per 200~
500mL collects a stream part, and TLC detections are carried out to effluent, merges identical stream part, obtains 4 Arius part (FrB20.1-
FrB20.4) and 2 Arius parts (FrB24.1-FrB24.2), removal of solvent under reduced pressure, obtain second and cross post part；
Phase III：It is 100 that post part FrB20.4 stream part, i.e. effluent volume ratio will be crossed for second:0 stream part；
FrB24.1 stream parts, i.e. effluent volume ratio is 70:30 stream part, is splined on respectively high performance liquid chromatography separation post, isolated
Benzofuran glycosides compound.Wherein, high performance liquid chromatography separation post for Jiangsu Chinese nation Megres C18 posts, high-efficient liquid phase color
Spectrum separation is to use differential refraction detector, using the acetic acid of methanol -0.5% water system or the acetic acid water system of acetonitrile -0.5% as flowing
Phase, by 3~6mL/min isocratic elution is carried out.
The volume ratio of methanol, water and acetic acid is respectively 45 in the acetic acid water system of methanol -0.5%：55：0.5 and 35：65：
0.5；The volume ratio of acetonitrile, water and acetic acid is respectively 42 in the acetic acid water system of acetonitrile -0.5%：58：0.5 and 31：69：0.5.
New benzofuran glycosides compound in the present invention can be applied in anti-inflammatory drug and/or health product is prepared.
Embodiment 1
1. benzofuran glycosides compound is isolated and purified
1) Kan Kula dry root 6kg are taken, with the methanol heating and refluxing extraction 3 times of 30L, 2 hours every time, now, and methanol
Volume be 5 times of quality of Xingan's Radix Angelicae Pubescentiss dry root, merge methanol extract liquid decompression and solvent recovery, obtain total extractum；
2) total extractum is suspended in 3 times of volume water, with petroleum ether equal-volume extraction 4 times, then Jing chloroforms successively, second
Acetoacetic ester and n-butyl alcohol equal-volume extraction, every kind of organic solvent is extracted 4 times, and organic layer normal pressure or vacuum distillation are removed after solvent
Respectively obtain petroleum ether layer, chloroform layer, ethyl acetate layer, n-butanol layer and remaining water layer.
3) n-butanol extraction layer 100g is taken, silica gel column chromatography is splined on, using chloroform-methanol system as eluent, by body
Product is (0 than (v/v):100)~(100:0) gradient elution is carried out, is collected once per 500mL, air-distillation recycling design, TLC
The identical stream part of merging is known in inspection, and 30 stream parts are obtained, and is respectively designated as FrB1-FrB30, takes FrB20 stream parts therein and FrB24
Stream part, i.e. effluent volume ratio respectively 10:1 and 8:1 stream part, as first time post part is crossed.
Wherein, the inverted silica gel column chromatography of FrB20 stream parts, with methanol-water system as eluent, by volume (v/v)
For 0:100~100:0 gradient elution, collects once, removal of solvent under reduced pressure per 200mL, and the identical stream part of merging is known in TLC inspections, obtains 4
Individual Arius part, is respectively designated as FrB20.1-FrB20.4.Then FrB20.4 stream parts, i.e. effluent volume ratio are 100:0 stream
Part, with half preparative high-performance liquid chromatographic instrument, using Megres C18 chromatographic columns, using methanol and 0.5% acetic acid water as flowing
Equality eluting (45:55), flow velocity is 3mL/min, obtains benzofuran glycosides compound 1 (retention time is 27min).
The inverted silica gel column chromatography of FrB24 stream parts, with methanol-water gradient elution (30:70,45:55,70:30,100:0,
V/v), collect once per 200mL, removal of solvent under reduced pressure, the identical stream part of merging is known in TLC inspections, obtains 2 Arius parts and is respectively designated as
FrB24.1-FrB24.2.Then FrB24.1 stream parts, i.e. effluent volume ratio are 70:30 stream part, with half efficient liquid phase is prepared
Chromatograph, using Megres C18 chromatographic columns, using methanol and 0.5% acetic acid water as mobile phase isocratic elution (35:65), flow
Speed is 3mL/min, obtains benzofuran glycosides compound 2 (retention time is 20min).
4) present invention is identified by physicochemical constant and Modern spectroscopy technological means (HR-ESI-MS, 1D-NMR, 2D-NMR)
The structure of compound, compound 1 is 6-methoxycarbonylethyl-benzofuran-5-O- β-D-xylopyranosyl
(1 → 2)-β-D-glucopyranoside, compound 2 be 6-hydroxycarbonylethyl-benzofuran-5-O- β-
D-xylopyranosyl(1→2)-β-D-glucopyranoside.The Structural Identification process of compound 1 and 2 is as described below.
2. the Structural Identification of new benzofuran glycosides compound
(1) Structural Identification of noval chemical compound 1
Compound 1 is yellow oil, and its HR-ESIMS shows that quasi-molecular ion peak is [M+Na]+at m/z
537.1584 (calcd 537.1579), it is thus determined that molecular formula is C23H30O13.Infrared spectrum shows hydroxyl group absorption band
(3403cm-1), alkyl absorption band (2933cm-1), carbonyl absorption band (1718cm-1) etc..Fig. 1's1In H NMR, four hydrogen matter
Subsignal δH7.64 (1H, d, J=2.2Hz), 7.18 (1H, m), 7.11 (1H, d, J=8.4Hz), 7.19 (1H m) shows point
There is disubstituted benzofuran structure in son, and two substituent groups are in the replacement of ortho position two.Fig. 2's13In C NMR, except one
Outer (the δ of individual carbonylC174.8), also eight fragrant carbon signals, including four methines, four quaternary carbons, the neighbour with aforementioned supposition
Position disubstituted benzofuran structure fragment is consistent.Compose with reference to the DEPT of Fig. 3, Fig. 4's1H-1The HMQC of H COSY spectrums and Fig. 5, two groups
Multiplet proton signal δH3.30 (1H, m), 3.01 (1H, m), δH2.72 (1H, m), 2.65 (1H, m) with their corresponding carbon
Signal (δC25.2 and δC34.9) show there are two methylene being joined directly together in molecule.In the HMBC spectrums of Fig. 6, this two
Individual methylene and a carbonyl carbon (δC174.8) it is related, and this carbonyl carbon is connected (δ with a methoxyl groupH3.68, δC
50.6).1H and13In C H NMR spectroscopies, two anomeric proton signal δH5.09 (1H, d, J=7.6Hz) and 4.79 (1H, d, J
=7.3Hz) and their corresponding carbon signal (δC101.8, δC104.5) show there are two sugared units in molecule, use after acid hydrolysis
GC Analysis and Identification both sugar are D-Glucose and D- xyloses, and the two sugared end group configurations are beta comfiguration, because their end
The coupling constant of substrate is 7.6Hz and 7.3Hz.The position of substitution and the order of connection of substituent group and sugar passes through integration analysis1H–1H-COSY, HMQC and HMBC are determined.
Therefore, the compound identification is：6-methoxycarbonylethyl-benzofuran-5-O-β-D-
Xylopyranosyl (1 → 2)-β-D-glucopyranoside, i.e. 6- dion e-benzofuran -5-O- β-D-
Xylopyranose-(1 → 2)-β-D- pyranglucoside, it is a noval chemical compound with new aglycon for having no document report.
Its structure sees below formula：
Nuclear magnetic data is shown in Table 1.
(2) Structural Identification of noval chemical compound 2
Compound 2 is yellow oil, and its HR-ESIMS shows that quasi-molecular ion peak is [M+Na]+at m/z
523.1427 (calcd 523.1422), determine that molecular formula is C22H28O13.The hydrogen spectrum and the carbon modal data and chemical combination of Fig. 8 of its Fig. 7
Thing 1 is quite similar, and a methoxyl group signal has simply been lacked in hydrogen spectrum, therefore speculates that compound 2 is the structure of acid rather than methyl ester,
This is consistent with molecular weight.Integration analysis Fig. 9's1H–1H-COSY, HMQC and HMBC are composed, and determine that compound 2 is 6-
hydroxycarbonylethyl-benzofuran-5-O-β-D-xylopyranosyl(1→2)-β-D-
Glucopyranoside, i.e. 6- dion e-benzofuran -5-O- β-D- xylopyranoses-(1 → 2)-β-D- pyrans
Glucoside, is a noval chemical compound for having no document report.The structural formula of compound 2 is as follows：
Compound 1 and 2 in the present invention of table 11H NMR and13C NMR datas
Further do pharmacologically active detection to the compound 1 and 2 of separated identification in the present invention below.
3. extracorporeal anti-inflammatory experiment
Experimental technique：
RAW264.7 cells are inoculated in 96 orifice plates, after culture 24h LPS (lipopolysaccharide) is added.Then different component is other
The sample (the compounds of this invention 1 and 2, positive drug indomethacin) of variable concentrations, blank group is added to add equal-volume
vehicle.50 μ L cell culture fluid mixing equal-volume Griess reagents I are added in above-mentioned 96 orifice plate, are subsequently added into Griess examinations
Agent II.Microplate reader determines OD value of each group sample under 546nm wavelength, calculates NO using following equation and produces suppression ratio, is used in combination
Half-inhibition concentration (the IC of the tested sample of SPSS computed in software50Value, μM).
NO suppression ratio (%)=[1-(sample sets/blank group)] × 100%.
Experimental result：
The compounds of this invention 1 of table 2 and the cell RAW264.7 of 2 pairs of LPS stimulations produce the inhibitory action of NO
Interpretation of result：The as shown by data of table 2, the compound 1 and 2 in the present invention is acted on good extracorporeal anti-inflammatory, right
Lipopolysaccharide LPS stimulates mice RAW264.7 cells generation NO to have good inhibiting effect, its half-inhibition concentration IC50Respectively
28.6 ± 1.6 μM, 32.9 ± 2.1 μM, it is superior to 39.5 ± 3.2 μM of positive control medicine indomethacin.In illustrating the present invention
Benzofuran glycosides compound 1 and 2 have preferable antiinflammatory action.
Embodiment 2
1) Kan Kula dry root 10kg are taken, is extracted 3 times with 70% alcohol heating reflux of 40L, 2 hours every time, merging was carried
Liquid and decompression and solvent recovery are taken, total extractum is obtained；
2) total extractum is suspended in 4 times of amount volume of water, after being first extracted with ethyl acetate 3 times, then uses n-butyl alcohol equal-volume
Extraction 4 times, is removed under reduced pressure after organic solvent and respectively obtains ethyl acetate layer and n-butanol layer.
3) n-butanol extraction layer 200g is taken, silica gel column chromatography is splined on, using chloroform-methanol system as eluent, by body
Product is 100 than (v/v):0~0:100 carry out gradient elution, collect once per 800mL, air-distillation recycling design, and TLC inspections are known
Merge identical stream part, 30 stream parts are obtained, be respectively designated as FrB1-FrB30, take FrB20 stream parts therein and FrB24 streams
Part, i.e. effluent volume ratio respectively 10:1 and 8:1 stream part, as first time post part is crossed.
Wherein, the inverted silica gel column chromatography of FrB20 stream parts, with methanol-water system as eluent, by volume (v/v)
For 100:0~0:100 carry out gradient elution, collect once per 400mL, removal of solvent under reduced pressure, and the identical stream part of merging is known in TLC inspections,
4 Arius parts are obtained, FrB20.1-FrB20.4 is respectively designated as.Then FrB20.4 stream parts, i.e. effluent volume ratio are 100%
Methanol-eluted fractions, with half preparative high-performance liquid chromatographic instrument, using Megres C18 chromatographic columns, with acetonitrile and 0.5% acetic acid
Water is used as mobile phase isocratic elution (42:58), flow velocity is 3mL/min, and (retention time is to obtain benzofuran glycosides compound 1
25min)。
The inverted silica gel column chromatography of FrB24 stream parts, is 100 with methanol-water volume ratio (v/v):0~0:100 carry out gradient
Eluting, collects once, removal of solvent under reduced pressure per 400mL, and the identical stream part of merging is known in TLC inspections, obtains 2 Arius parts and is respectively
FrB24.1-FrB24.2.Then FrB24.1 stream parts, i.e. effluent volume ratio are 70:30 stream part, with half efficient liquid phase is prepared
Chromatograph, using Megres C18 chromatographic columns, using acetonitrile and 0.5% acetic acid water as mobile phase isocratic elution (31:69), flow
Speed is 3mL/min, obtains benzofuran glycosides compound 2 (retention time is 18min).
Embodiment 3
1) Kan Kula dry root 10kg are taken, with the water heating and refluxing extraction 1 time of 30L, 4 hours every time, united extraction liquid was simultaneously
Decompression and solvent recovery, obtains total extractum；
2) total extractum is suspended in 2 times of amount volume of water, after being first extracted with ethyl acetate 3 times, then uses n-butyl alcohol equal-volume
Extraction 6 times, is removed under reduced pressure after organic solvent and respectively obtains ethyl acetate layer and n-butanol layer.
3) n-butanol extraction layer 200g is taken, silica gel column chromatography is splined on, using chloroform-methanol system as eluent, by body
Product is 100 than (v/v):0~0:100 carry out gradient elution, collect once per 800mL, air-distillation recycling design, and TLC inspections are known
Merge identical stream part, 30 stream parts are obtained, be respectively designated as FrB1-FrB30, take FrB20 stream parts therein and FrB24 streams
Part, i.e. effluent volume ratio respectively 10:1 and 8:1 stream part, as first time post part is crossed.
Wherein, the inverted silica gel column chromatography of FrB20 stream parts, with methanol-water system as eluent, by volume (v/v)
For 100:0~0:100 carry out gradient elution, collect once per 500mL, removal of solvent under reduced pressure, and the identical stream part of merging is known in TLC inspections,
4 Arius parts are obtained, FrB20.1-FrB20.4 is respectively designated as.Then FrB20.4 stream parts, i.e. effluent volume ratio are 100%
Methanol-eluted fractions, with half preparative high-performance liquid chromatographic instrument, using Megres C18 chromatographic columns, with acetonitrile and 0.5% acetic acid
Water is used as mobile phase isocratic elution (42:58), flow velocity is 3mL/min, and (retention time is to obtain benzofuran glycosides compound 1
25min)。
The inverted silica gel column chromatography of FrB24 stream parts, with methanol-water, by volume (v/v) is 100:0~0:100 carry out ladder
Degree eluting, collects once, removal of solvent under reduced pressure per 400mL, and TLC inspection knowledges merge identical stream part, obtain 2 Arius parts and are respectively
FrB24.1-FrB24.2.Then FrB24.1 stream parts, i.e. effluent volume ratio are 70:30 stream part, with half efficient liquid phase is prepared
Chromatograph, using Megres C18 chromatographic columns, using acetonitrile and 0.5% acetic acid water as mobile phase isocratic elution (31:69), flow
Speed is 3mL/min, obtains benzofuran glycosides compound 2 (retention time is 18min).
Embodiment 4
1) Kan Kula dry root 10kg are taken, is extracted 6 times with 10% alcohol heating reflux of 80L, 1 hour every time, merging was carried
Liquid and decompression and solvent recovery are taken, total extractum is obtained；
2) by total extractum be suspended in 5 times amount volume of water in, first with petroleum ether extraction 2 times after, then successively use chloroform, acetic acid second
Ester and n-butyl alcohol equal-volume extraction 1 time, are removed under reduced pressure after organic solvent and respectively obtain ethyl acetate layer and n-butanol layer.
3) n-butanol extraction layer 200g is taken, silica gel column chromatography is splined on, using chloroform-methanol system as eluent, by body
Product is 100 than (v/v):0~0:100 carry out gradient elution, collect once per 300mL, air-distillation recycling design, and TLC inspections are known
Merge identical stream part, 30 stream parts are obtained, be respectively designated as FrB1-FrB30, take FrB20 stream parts therein and FrB24 streams
Part, i.e. effluent volume ratio respectively 10:1 and 8:1 stream part, as first time post part is crossed.
Wherein, the inverted silica gel column chromatography of FrB20 stream parts, with methanol-water system as eluent, by volume (v/v)
For 100:0~0:100 carry out gradient elution, collect once per 300mL, removal of solvent under reduced pressure, and the identical stream part of merging is known in TLC inspections,
4 Arius parts are obtained, FrB20.1-FrB20.4 is respectively designated as.Then FrB20.4 stream parts, i.e. effluent volume ratio are 100%
Methanol-eluted fractions, with half preparative high-performance liquid chromatographic instrument, using Megres C18 chromatographic columns, with acetonitrile and 0.5% acetic acid
Water is used as mobile phase isocratic elution (42:58), flow velocity is 3mL/min, and (retention time is to obtain benzofuran glycosides compound 1
25min)。
The inverted silica gel column chromatography of FrB24 stream parts, with methanol-water, by volume (v/v) is 100:0~0:100 carry out ladder
Degree eluting, collects once, removal of solvent under reduced pressure per 400mL, and TLC inspection knowledges merge identical stream part, obtain 2 Arius parts and are respectively
FrB24.1-FrB24.2.Then FrB24.1 stream parts, i.e. effluent volume ratio are 70:30 stream part, with half efficient liquid phase is prepared
Chromatograph, using Megres C18 chromatographic columns, using acetonitrile and 0.5% acetic acid water as mobile phase isocratic elution (31:69), flow
Speed is 3mL/min, obtains benzofuran glycosides compound 2 (retention time is 18min).
Embodiment 5
1) Kan Kula dry root 10kg are taken, is extracted 4 times with 95% alcohol heating reflux of 35L, 3 hours every time, merging was carried
Liquid and decompression and solvent recovery are taken, total extractum is obtained；
2) by total extractum be suspended in 1 times amount volume of water in, first with petroleum ether extraction 1 time after, then successively use chloroform, acetic acid second
Ester and n-butyl alcohol equal-volume extraction 2 times, are removed under reduced pressure after organic solvent and respectively obtain ethyl acetate layer and n-butanol layer.
3) n-butanol extraction layer 200g is taken, silica gel column chromatography is splined on, using chloroform-methanol system as eluent, by body
Product is 100 than (v/v):0~0:100 carry out gradient elution, collect once per 600mL, air-distillation recycling design, and TLC inspections are known
Merge identical stream part, 30 stream parts are obtained, be respectively designated as FrB1-FrB30, take FrB20 stream parts therein and FrB24 streams
Part, i.e. effluent volume ratio respectively 10:1 and 8:1 stream part, as first time post part is crossed.
Wherein, the inverted silica gel column chromatography of FrB20 stream parts, with methanol-water system as eluent, by volume (v/v)
For 100:0~0:100 carry out gradient elution, collect once per 200mL, removal of solvent under reduced pressure, and the identical stream part of merging is known in TLC inspections,
4 Arius parts are obtained, FrB20.1-FrB20.4 is respectively designated as.Then FrB20.4 stream parts, i.e. effluent volume ratio are 100%
Methanol-eluted fractions, with half preparative high-performance liquid chromatographic instrument, using Megres C18 chromatographic columns, with methanol and 0.5% acetic acid
Water is used as mobile phase isocratic elution (45:55), flow velocity is 3mL/min, and (retention time is to obtain benzofuran glycosides compound 1
27min)。
The inverted silica gel column chromatography of FrB24 stream parts, with methanol-water, by volume (v/v) is 100:0~0:100 carry out ladder
Degree eluting, collects once, removal of solvent under reduced pressure per 400mL, and TLC inspection knowledges merge identical stream part, obtain 2 Arius parts and are respectively
FrB24.1-FrB24.2.Then FrB24.1 stream parts, i.e. effluent volume ratio are 70:30 stream part, with half efficient liquid phase is prepared
Chromatograph, using Megres C18 chromatographic columns, using methanol and 0.5% acetic acid water as mobile phase isocratic elution (35:65), flow
Speed is 3mL/min, obtains benzofuran glycosides compound 2 (retention time is 20min).
Experiment and its experimental result, show that the benzofuran glycosides compound in the present invention has good body with reference to more than
The effect of outer antiinflammatory, is expected to exploitation for new anti-inflammatory drug；Or for preparing the health food of prevention and treatment inflammation.
Comparative example 1
By step 5) in mobile phase change the identical mobile phase of other mobile phases either other ratios into (or chromatographic column replaced
Change common reverse phase silica gel chromatographic column into), other steps and condition it is same as Example 1.It was found that benzofuran glycosides cannot be isolated
Class compound.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention not by above-described embodiment
Limit, other any spirit without departing from the present invention and the change, modification, replacement made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (10)
1. benzofuran glycosides compound, it is characterised in that：Its molecular formula is C23H30O13Or C22H28O13。
2. benzofuran glycosides compound according to claim 1, it is characterised in that：Its general structure is：
Wherein, R is-H or-CH3。
3. the preparation method of benzofuran glycosides compound, it is characterised in that：Comprise the following steps：
1) taking the dry root of Xingan's Radix Angelicae Pubescentiss carries out reflux, extract, several times, by extracting solution concentrating under reduced pressure, obtains total extractum or concentration
Liquid；
2) total extractum is suspended in water, obtains extractum liquid, extractum liquid or concentrated solution are separated Jing after ethyl acetate is extracted several times
Go out organic layer, remaining water layer passes through n-butanol extraction rear combining extraction liquid several times, n-butyl alcohol is removed under reduced pressure and obtains n-butyl alcohol extraction
Take layer；
3) n-butanol extraction layer is splined on into silicagel column, using chloroform-methanol system as eluent, by volume (100:0)~
(0:100) gradient elution is carried out, effluent is detected, be 10 by effluent volume ratio:1 stream part merges, and removes solvent,
Obtain sample a1；It is 8 by effluent volume ratio:1 stream part merges, and removes solvent, obtains sample b1；
4) sample a1 is splined on into reversed-phase silica gel chromatography post, using methanol-water system as eluent, by volume (0:100)~
(100:0) gradient elution is carried out, is 100 by effluent volume ratio:0 stream part merges, and removes solvent, obtains sample a2；
Sample b1 is splined on into reversed-phase silica gel chromatography post, using methanol-water system as eluent, by volume (0:100)~
(100:0) gradient elution is carried out, is 70 by effluent volume ratio:30 stream part merges, and removes solvent, obtains sample b2；
5) sample a2 and sample b2 are splined on into respectively high performance liquid chromatography separation post, with mobile phase isocratic elution is carried out, obtained
Benzofuran glycosides compound.
4. the preparation method of benzofuran glycosides compound according to claim 3, it is characterised in that：Step 1) backflow
Using the ethanol of methanol, water or volume fraction 10~95% as extractant, the dry root and extractant of Xingan's Radix Angelicae Pubescentiss in extraction
Mass volume ratio be 1kg：(1~8) L；When the ethanol that extractant is methanol or volume fraction 10~95%, by carrying for obtaining
Take solvent recovery in liquid and obtain total extractum；When extractant be water when, by the volume concentration of extracting solution to ten/6ths/
One, obtain concentrated solution.
5. the preparation method of benzofuran glycosides compound according to claim 3, it is characterised in that：Step 1) middle extraction
Number of times is 1~6 time, every time 1~4 hour.
6. the preparation method of benzofuran glycosides compound according to claim 3, it is characterised in that：Step 2) in always soak
The volume ratio of cream and water is 1:1~1:5.
7. the preparation method of benzofuran glycosides compound according to claim 3, it is characterised in that：Step 2) in extractum
Liquid or concentrated solution are directly over ethyl acetate extraction；Or first successively Jing after petroleum ether and chloroform extraction, then through ethyl acetate
Extraction；Every time extraction is equal-volume extraction；Every kind of solvent extracts respectively 1~6 time.
8. the preparation method of benzofuran glycosides compound according to claim 3, it is characterised in that：Step 3) in gradient
After eluting per 300~800mL collect a stream part；Step 4) in a stream part is collected per 200~500mL after gradient elution；Step
It is rapid 3) and step 4) in effluent be to carry out TLC detections；Step 5) in isocratic elution flow velocity be 3~6mL/min.
9. the preparation method of benzofuran glycosides compound according to claim 3, it is characterised in that：Step 5) middle flowing
It is mutually the acetic acid of methanol -0.5% water system or the acetic acid water system of acetonitrile -0.5%；When carrying out isocratic elution to sample a2, employing
The volume ratio of methanol, water and acetic acid is 45 in the acetic acid water system of methanol -0.5%：55：0.5, the acetic acid water of acetonitrile -0.5% of employing
The volume ratio of acetonitrile, water and acetic acid is 42 in system：58：0.5, the benzofuran glycosides compound molecular formula for obtaining is
C23H30O13；When carrying out isocratic elution to sample a2, the body of methanol, water and acetic acid in the acetic acid water system of methanol -0.5% of employing
Product is than being 35：65：0.5, the volume ratio of acetonitrile, water and acetic acid is for 31 in the acetic acid water system of acetonitrile -0.5% for adopting：69：0.5,
The benzofuran glycosides compound molecular formula for obtaining is C22H28O13。
10. answering in terms of benzofuran glycosides compound as claimed in claim 1 is in anti-inflammatory drug and/or health product is prepared
With.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611228845.1A CN106674311B (en) | 2016-12-27 | 2016-12-27 | A kind of benzofuran glycosides compound and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611228845.1A CN106674311B (en) | 2016-12-27 | 2016-12-27 | A kind of benzofuran glycosides compound and its preparation method and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106674311A true CN106674311A (en) | 2017-05-17 |
| CN106674311B CN106674311B (en) | 2019-02-05 |
Family
ID=58872997
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611228845.1A Expired - Fee Related CN106674311B (en) | 2016-12-27 | 2016-12-27 | A kind of benzofuran glycosides compound and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106674311B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112851612A (en) * | 2021-01-30 | 2021-05-28 | 河南中医药大学 | Active compound extracted from burdock leaves and having cholesterol reducing effect, and preparation method and application thereof |
| CN112920148A (en) * | 2021-01-30 | 2021-06-08 | 河南中医药大学 | New compound NBY-16 extracted from folium Arctii and having antiinflammatory activity, and its preparation method and application |
| WO2021261844A1 (en) * | 2020-06-26 | 2021-12-30 | 한국과학기술연구원 | Anti-obesity, anti-dyslipidemia, antioxidant or anti-inflammatory composition containing angelica dahurica extract |
| CN114644674A (en) * | 2020-12-21 | 2022-06-21 | 广西大学 | Method for separating eclipta saponin I from ovate leaf holly bark |
| CN115703753A (en) * | 2021-08-16 | 2023-02-17 | 江苏康缘药业股份有限公司 | Benzofuran type derivative and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1633432A (en) * | 2002-04-08 | 2005-06-29 | 科学与工业研究委员会 | Novel glucopyranoside, process for isolation thereof, pharmaceutical composition containing same and use thereof |
| CN1830976A (en) * | 2005-03-08 | 2006-09-13 | 成都地奥制药集团有限公司 | Extractive of total benzofuran of benzoin plants, prepn. method and use thereof |
| US20070148268A1 (en) * | 2005-12-23 | 2007-06-28 | Hong Kong Jockey Club Institute Of Chinese Medicine Limted | Chemical compounds and their use as therapeutic agent and means of quality assessment of herbal medicines |
| CN101502507A (en) * | 2008-02-04 | 2009-08-12 | 成都地奥九泓制药厂 | Egonol type benzofuran and novel use in pharmacy of glycosides thereof |
-
2016
- 2016-12-27 CN CN201611228845.1A patent/CN106674311B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1633432A (en) * | 2002-04-08 | 2005-06-29 | 科学与工业研究委员会 | Novel glucopyranoside, process for isolation thereof, pharmaceutical composition containing same and use thereof |
| CN1830976A (en) * | 2005-03-08 | 2006-09-13 | 成都地奥制药集团有限公司 | Extractive of total benzofuran of benzoin plants, prepn. method and use thereof |
| US20070148268A1 (en) * | 2005-12-23 | 2007-06-28 | Hong Kong Jockey Club Institute Of Chinese Medicine Limted | Chemical compounds and their use as therapeutic agent and means of quality assessment of herbal medicines |
| CN101502507A (en) * | 2008-02-04 | 2009-08-12 | 成都地奥九泓制药厂 | Egonol type benzofuran and novel use in pharmacy of glycosides thereof |
Non-Patent Citations (2)
| Title |
|---|
| CHUN-FENG QIAO ET AL.: "Psoralenoside and Isopsoralenoside, Two New Benzofuran Glycosides from Psoralea corylifolia", 《CHEM. PHARM. BULL.》 * |
| KAZUKO YOSHIKAWA ET AL.: "A Benzofuran Glycoside and an Acetylenic Acid from the Fungus Laetiporus sulphureus var. miniatus", 《CHEM. PHARM. BULL.》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021261844A1 (en) * | 2020-06-26 | 2021-12-30 | 한국과학기술연구원 | Anti-obesity, anti-dyslipidemia, antioxidant or anti-inflammatory composition containing angelica dahurica extract |
| KR20220000640A (en) * | 2020-06-26 | 2022-01-04 | 한국과학기술연구원 | A composition for anti-obesity, anti-dyslipidemia, anti-oxidation or anti-inflammation comprising extracts of heracleum dissectum |
| KR102466616B1 (en) * | 2020-06-26 | 2022-11-15 | 한국과학기술연구원 | A composition for anti-obesity, anti-dyslipidemia, anti-oxidation or anti-inflammation comprising extracts of heracleum dissectum |
| CN114644674A (en) * | 2020-12-21 | 2022-06-21 | 广西大学 | Method for separating eclipta saponin I from ovate leaf holly bark |
| CN114644674B (en) * | 2020-12-21 | 2023-05-12 | 广西大学 | Method for separating eclipta saponin I from holly bark |
| CN112851612A (en) * | 2021-01-30 | 2021-05-28 | 河南中医药大学 | Active compound extracted from burdock leaves and having cholesterol reducing effect, and preparation method and application thereof |
| CN112920148A (en) * | 2021-01-30 | 2021-06-08 | 河南中医药大学 | New compound NBY-16 extracted from folium Arctii and having antiinflammatory activity, and its preparation method and application |
| CN112851612B (en) * | 2021-01-30 | 2023-11-10 | 河南中医药大学 | Active compound extracted from burdock leaves and capable of reducing cholesterol, and preparation method and application thereof |
| CN112920148B (en) * | 2021-01-30 | 2023-11-14 | 河南中医药大学 | Novel compound NBY-16 extracted from burdock leaves and having anti-inflammatory activity, and preparation method and application thereof |
| CN115703753A (en) * | 2021-08-16 | 2023-02-17 | 江苏康缘药业股份有限公司 | Benzofuran type derivative and preparation method and application thereof |
| CN115703753B (en) * | 2021-08-16 | 2024-02-06 | 江苏康缘药业股份有限公司 | Benzofuran derivative and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106674311B (en) | 2019-02-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106674311B (en) | A kind of benzofuran glycosides compound and its preparation method and application | |
| Jiang et al. | Chemical constituents of Cynanchum wilfordii and the chemotaxonomy of two species of the family Asclepiadacease, C. wilfordii and C. auriculatum | |
| CN102247393B (en) | Preparation method of oleanolic acid saponin component and application thereof | |
| CN109942649B (en) | Indole glycoside compound and extraction and separation method and application thereof | |
| CN101648937B (en) | Isoflavones compound and preparation and application thereof | |
| CN106674309B (en) | Tonka bean camphor glycosides compounds and its preparation method and application | |
| CN103169727A (en) | General-flavonoid compound in chionanthus as well as preparation method and application thereof | |
| CN102942605B (en) | The preparation method of iridoid and application thereof in Jasminum lanceolarium Roxb. | |
| CN102178688A (en) | Preparation method of ivy saponins ingredient and application of the ingredient to resisting tumors | |
| CN102772501A (en) | Rheum emodi Wall. extract and its preparing method | |
| CN106674299B (en) | Carbene glycosides compound and its preparation method and application | |
| CN103739660B (en) | A kind of compound, its extracting method, its application preparing antitumor drug and antitumor drug of preparation thereof | |
| CN104059123B (en) | The antitumor drug of a kind of Sasanguasaponin compound, its preparation method, application and preparation thereof | |
| CN103113439A (en) | Method for preparing kaempferol-3-O-Beta-D-glucuronide in euphorbia sororia | |
| CN103833818B (en) | The antitumor drug of a kind of Sasanguasaponin compound, its preparation method, application and preparation thereof | |
| CN106565444A (en) | Extraction method and application of phenanthrene compounds from overground part of Chinese yam | |
| CN107722087B (en) | Gynostemma pentaphylla flavonoid compound, preparation method thereof and application thereof in antitumor drugs | |
| CN103113196B (en) | Glechoma longituba phenol, and preparation method and application thereof | |
| CN103739657B (en) | A kind of Sasanguasaponin compound, its preparation method, the antitumor drug applying and prepare | |
| CN102188502B (en) | Extraction method and composition of common souliea rhizome total saponins with anti-tumor effect | |
| CN103665090B (en) | Ilex cornuta saponin compound, its preparation method and application | |
| CN109232602A (en) | A kind of linear furocoumarins class compound and its preparation method and application | |
| CN106854227B (en) | Benzenpropanoic acid glycosides compound and its preparation method and application | |
| CN102349937B (en) | Preparation method of ixeris sonchifolia hance extractive | |
| CN103169724A (en) | Preparation method and application of oleanolic acid trisaccharide saponin component |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190205 Termination date: 20211227 |