CN105646151B - A kind of purposes of acetylene compound and preparation method thereof and the compound - Google Patents
A kind of purposes of acetylene compound and preparation method thereof and the compound Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C33/00—Unsaturated compounds having hydroxy or O-metal groups bound to acyclic carbon atoms
- C07C33/04—Acyclic alcohols with carbon-to-carbon triple bonds
- C07C33/048—Acyclic alcohols with carbon-to-carbon triple bonds with double and triple bonds
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- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
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- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
- C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
- C07C29/86—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment by liquid-liquid treatment
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- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
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- C07C67/58—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/02—Esters of acyclic saturated monocarboxylic acids having the carboxyl group bound to an acyclic carbon atom or to hydrogen
- C07C69/12—Acetic acid esters
- C07C69/14—Acetic acid esters of monohydroxylic compounds
- C07C69/145—Acetic acid esters of monohydroxylic compounds of unsaturated alcohols
Abstract
The present invention provides a kind of acetylene compounds and preparation method thereof and the purposes of the compound, belong to acetylene compound and extract Technique of Chinese Medicine Efficacious Ingredient field.Specifically extracted new compounds i isolated from the dry root of Umbelliferae Bupleunim. L radix bupleuri (Radix Bupleuri), II, IV and reported compound III.External monoamine neurotransmitter reuptake Inhibition test is carried out to above compound, the result shows that chemical compounds I, the reuptake of II, III and IV pair of norepinephrine (NA) are respectively provided with inhibiting effect, inhibition is equivalent to positive control medicine desipramine, and different degrees of inhibiting effect is generated to the reuptake of 5 hydroxytryptophans (5 HT).Above-mentioned four kinds of compounds have antidepressant activity, are expected to drug or its pro-drug of the exploitation to treat and prevent antidepression relevant disease.
Description
Technical field
The present invention relates to acetylene compounds and extract Technique of Chinese Medicine Efficacious Ingredient field, and in particular to a kind of acetylene compound
With the method for separation acetylene compound from radix bupleuri and the purposes of the acetylene compound.
Background technology
Depression is a kind of common mental disease for having the characteristics that high incidence, high recurrence rate and height and disabling.Modern times doctor
Learn research shows that, depression complicated mechanism, induced factor is more, and clinical disease is various.Most experts and scholars think, depressed
Monoamine hypothesis, that is, depression of disease is due to human brain monoamine neurotransmitter, caused by 5-HT, NA or DA defect.Present clinically institute
Main antidepressant is by the way that the reuptake of synaptic cleft 5-HT and NA is inhibited to increase monoamine neurotransmitter in synaptic cleft
Concentration.But synthesizing antidepressant, there are the shortcomings of antidepression spectrum is narrow, and onset time is slow and toxic side effect is big.Therefore, it is domestic
It is outer to start to pay attention to develop from conventional medicament and natural drug and drug of the exploitation with antidepressant activity.
Xiaoyao San is classics recipe, has the applicating history of nearly one thousand years so far.Modern clinic has proven to it with pharmacological research
With apparent antidepressant effect, and toxic side effect is not found.This seminar early period to Xiaoyao San each position carried out it is external and
Internal pharmacology activity research has filtered out petroleum ether active site, finds to be substantially better than former compound in terms of its antidepression curative effect, very
To the activity for being more than Western medicine, patent applied for (ZL201010110283.7), but still there are material base it is indefinite the problems such as.
This research, using serum drug chemistry as guidance, carries out the chemical composition of monarch drug in a prescription radix bupleuri based on the research to effective substance
Research finds that the content of wherein polyacetylene compound is higher, and can enter blood, may be antidepressant active ingredient, therefore right
It carries out in-depth study.
At present, polyyne constituents isolated from other plant once have significant activity by researcher's report.Such as
The effect for the antitumor cell that panaxytiol has；Known more than 20 kind polyacetylene class compound, which has, in rhizoma atractylodis prevents stomach from damaging
The effects that wound, anti-inflammatory, cholagogic and xanthine oxidase hinder.In recent years, correlative study person extracts from the plant of Bupleurum
The compound of isolated 17 kinds of polyacetylenes, wherein bupleurotoxin and acetylbupleurotoxin have significant neurotoxicity and cell
Toxic effect, other compounds have no the report of activity or toxicity.
The present invention extracts, petroleum ether in the research for radix bupleuri petroleum ether part antidepression active ingredient by ethyl alcohol
The method that extraction, chromatographic technique isolate and purify and activity experiment detection is combined, is finally obtained one with antidepressant activity
Group noval chemical compound, and by technologies such as high resolution NMRs, it is determined that the chemical constitution of each compound.Above compound is equal
With preferable antidepressant activity, the drug being made suitable for clinical treatment depression can be developed.If as the new chemical combination of guide
Object is conducted further research by structure of modification and to its structure-activity relationship, can synthesize a series of antidepressant and new
Pharmaceutical composition.
Invention content
The object of the present invention is to provide a kind of alkynes that the method for acetylene compound and this method are detached from radix bupleuri and is obtained
Class compound and its application in antidepressant is prepared.
A kind of acetylene compound provided by the invention, the structural formula with following I, II, III or IV：
The extracting method of above-mentioned acetylene compound I, II, III and IV, includes the following steps：
(1) prepared by radix bupleuri petroleum ether part：95% ethyl alcohol of radix bupleuri is impregnated, refluxing extraction, extracting solution is concentrated to give medicinal extract,
Medicinal extract petroleum ether extraction obtains radix bupleuri petroleum ether part；
(2) crude separation of radix bupleuri petroleum ether part：Will radix bupleuri petroleum ether part add in silicagel column in detach, with petroleum ether-
The mixed liquor elution of ethyl acetate different proportion, silica gel thin-layer is qualitative and efficient liquid phase tracer, collects petroleum ether-acetic acid second respectively
Ester ratio is 100：6 and 100：30 parts obtain crude product B and crude product C；
It uses a dry method on a sample, the mixed liquor of crude product B petroleum ether-ethyl acetate different proportions is eluted, collection petroleum ether-
Ethyl acetate ratio is 100：1 to 100：2 parts obtain the BA of crude product containing target compound.
Using wet method loading, the mixed liquor of crude product C petroleum ether-ethyl acetate different proportions is eluted, collection petroleum ether-
Ethyl acetate ratio is 100：30 parts obtain crude product CE.
Using wet method loading, the mixed liquor of crude product CE petroleum ether-ethyl acetate different proportions is eluted, collects oil
Ether-ethyl acetate ratio is 100：5 parts obtain the CEB of crude product containing target compound.
(3) it purifies：Crude product BA and crude product CEB by silica gel chromatographies column, ODS column chromatographies and Sephadex LH-20 are coagulated respectively
Rubber column gel column cleans, and is dissolved in 9：In 1 methanol-chloroform solvent, filter membrane filtration obtains sample BA-1 and CEB-3, spare.
(4) preparation of compound：Sample CEB-3 is taken, using high performance liquid chromatography, using methanol-water ratio as 88：12 are
Mobile phase is collected the chromatographic peak that retention time is 11.7-13.3min and 17.5-18.9min and is concentrated to dryness, obtains such as power respectively
Profit requires 1 chemical compounds I and compound ii.
Sample BA-1 is taken, using high performance liquid chromatography, using methanol-water ratio as 87.5：12.5 be mobile phase, is received respectively
The chromatographic peak for integrating retention time as 19.0-21.3min and 22.0-23.9min is concentrated to dryness, and obtains the compound III and chemical combination
Object IV.
Acetylene compound of the present invention is to extract to detach from bupleurum Chinese (Radix Bupleuri) dry root
It arrives, the methods of using NMR (1H-NMR and 13C-NMR) UV and MS, the structure of compound is identified, the present invention is from radix bupleuri stone
Isolated new compounds i, II, IV are the compounds do not reported in oily ether position, and compound III is the compound reported.This
Four kinds of compound specific chemical components, belong to low polarity component in plant, and have specific extraction preparation method, be expected to exploitation into
The specific new antidepressant of effective substance or antidepression pro-drug.
It is proved through experiment in vitro, chemical compounds I, II, III and IV can inhibit the reuptake behavior of NA, and inhibiting rate is right with the positive
It is suitable according to medicine desipramine；Though chemical compounds I, the reuptake behavior of II, III and IV couple of 5-HT are weaker than positive drug Fluoxetine hydrochloride,
Also there is different degrees of inhibiting effect.Prove that the acetylene compound can answer in the drug for preparing prevention and treatment depression
With.
Specific embodiment
Embodiment 1：The preparation of acetylene compound
(1) prepared by radix bupleuri petroleum ether part
Radix Bupleuri 5kg impregnates 12h with 95% ethyl alcohol, and three times, each 2h merges extracting solution and is concentrated to give leaching refluxing extraction
Cream, medicinal extract petroleum ether extraction obtain radix bupleuri petroleum ether part 150g.
(2) crude separation of radix bupleuri petroleum ether part
Above-mentioned radix bupleuri petroleum ether part sample is added in silica gel column chromatography, is by the use of petroleum ether-ethyl acetate as elution
System, uses petroleum ether successively：Ethyl acetate ratio is 100:0~0:100 carry out gradient elution.With silica G F254Thin layer chromatography board,
Petroleum ether-ethyl acetate detects fraction for development system, when thin-layer chromatography monitoring component has significant change, replaces next concentration and washes
De- gradient.With reference to high performance liquid chromatography tracking result, petroleum ether is collected：Ethyl acetate ratio is 100：6 and 100：30 parts, obtain
To crude product B and crude product C.
Crude product B is taken, is used a dry method on a sample, is placed in dring silicon rubber powder (200-300 mesh) chromatographic column (chromatographic column specification：Diameter
3cm, high 45cm), petroleum ether is used successively：Ethyl acetate ratio is 100：1~100：6 carry out gradient elution, with reference to efficient liquid phase
Chromatogram tracking is as a result, collect petroleum ether：Ethyl acetate ratio is 100：1 to 100：2 parts obtain crude product containing target compound
BA。
Crude product C is taken, using wet method loading, is placed in ODS columns and prepares, use petroleum ether successively：Ethyl acetate ratio is 100：6
~100：30 carry out gradient elution, with reference to high performance liquid chromatography tracking result, collect petroleum ether：Ethyl acetate ratio is 100：30
Part obtains crude product CE.
Crude product CE is taken, using wet method loading, is placed in ODS columns and prepares, use petroleum ether successively：Ethyl acetate ratio is 100：
3~100：15 carry out gradient elution, with reference to high performance liquid chromatography tracking result, collect petroleum ether：Ethyl acetate ratio is 100：5
Part obtains the CEB of crude product containing target compound.
(3) it purifies
Crude product BA is taken, through chloroform：Methanol (1:1) Sephadex LH-20 gel columns are detached, and obtain sample BA-1, so
Sample is dissolved in methanol afterwards：Chloroform (9:1) in solvent, 0.22 μm of membrane filtration is spare.
Crude product CEB is taken through chloroform：Methanol (1:1) Sephadex LH-20 gel columns are detached, and obtain sample CEB-3,
Then sample is dissolved in methanol：Chloroform (9:1) in solvent, 0.22 μm of membrane filtration is spare.
(4) preparation of compound
Membrane filtration sample CEB-3 in (3) is taken, using high performance liquid chromatography, with methanol：Water=88：12, it collects respectively
Retention time is that the chromatographic peak of 11.7-13.3min and 17.5-18.9min is concentrated to dryness, and obtains target compound I and compound
Ⅱ。
Membrane filtration sample BA-1 in (3) is taken, using high performance liquid chromatography, with methanol：Water=87.5：12.5 respectively
It collects the chromatographic peak that retention time is 19.0-21.3min and 22.0-23.9min to be concentrated to dryness, obtains compound III and compound
Ⅳ。
(5) structural confirmation of compound
This experimental compound structural confirmation, is belonged to using Correlated Spectroscopy, using Bruker 600MHz Avance III
NMR Spectrometer nuclear magnetic resonance spectrometers measure13H- and13C-NMR is composed, and Waters ACQUITY PULCTM PDA chromatographs measure
Ultraviolet spectra and Thermo Q Extractive LTQ Orbitrap Mass analyzer mass spectrographs measure ESI-MS, and join
The structure of kind of compound is identified according to document.
The Structural Identification result of compound is as follows：
1. chemical compounds I：
Chinese name：(2E, 8E, 10E) -2,8,10- pentadecane triolefin -4,6- diine -1- alcohol
English name：(2E,8E,10E)-pentadeca-2,8,10-trien-4,6-diyn-1-ol
Character：Colourless crystallization body (methanol)
UV(MeOH)λmax252nm 315nm 337nm
MS HR-ESI-MS m/z 215.1391[M+H]+
1H-NMR(CDCl3,/600MHz) and13C-NMR(CDCl3,/150MHz) attribution data is as shown in table 1.According to wave spectrum
Data and Scifinder retrievals, determine chemical compounds I as a noval chemical compound having not been reported, structural formula：
Molecular formula C15H18O, molecular weight 214.Hydrocarbon attribution data is shown in Table 1.
2. compound ii
Chinese name：(2E, 8E, 10E) -2,8,10- heptadecane triolefin -4,6- diine -1- alcohol
English name：(2E,8E,10E)-heptadeca-2,8,10-trien-4,6-diyn-1-ol
Character：Colourless crystallization body (methanol)
UV(MeOH)λmax253nm 315nm 337nm
MS HR-ESI-MS m/z 243.1704[M+H]+
1H-NMR(CDCl3,/600MHz) and13C-NMR(CDCl3,/150MHz) attribution data is as shown in table 1.According to wave spectrum
Data and Scifinder retrievals, determine compound ii as a noval chemical compound having not been reported, structural formula：
Molecular formula C17H22O, molecular weight 242.Hydrocarbon attribution data is shown in Table 1.
3. compound III
Chinese name：Three dilute -4,6- diines -1- alcohol ethyl esters of (2Z, 8E, 10E)-pentadecane
English name：(2Z,8E,10E)-pentadeca-2,8,10-trien-4,6-diyn-1-yl acetate
Character：Colourless crystallization body (methanol)
UV(MeOH)λmax 253nm 316nm 339nm
MS HR-ESI-MS m/z 257.1497[M+H]+
1H-NMR(CDCl3,/600MHz) and13C-NMR(CDCl3,/150MHz) attribution data is as shown in table 2.According to wave spectrum
Data and pertinent literature, determine compound III for (2Z, 8E, 10E)-pentadecane three it is dilute -4,6- diine -1- alcohol ethyl esters, structural formula：
Molecular formula C17H20O2, molecular weight 256.Hydrocarbon attribution data is shown in Table 2.
4. compounds Ⅳ
Chinese name：Three dilute -4,6- diines -1- alcohol ethyl esters of (2E, 8E, 10E)-pentadecane
English name：(2E,8E,10E)-pentadeca-2,8,10-trien-4,6-diyn-1-yl acetate
Character：Colourless crystallization body (methanol)
UV(MeOH)λmax 254nm 316nm 339nm
MS HREIMS m/z 257.1497[M+H]+
1H-NMR(CDCl3,/600MHz) and13C-NMR(CDCl3,/150MHz) attribution data is as shown in table 2.According to wave spectrum
Data and Scifinder retrievals, determine compounds Ⅳ as a noval chemical compound having not been reported, structural formula：
Molecular formula C17H20O2, molecular weight 256.Hydrocarbon attribution data is shown in Table 2.
Table 1 chemical compounds I and II13C-NMR (150MHz, CDCl3) and1H-NMR (600MHz, CDCl3) spectroscopic data
Table 2 compound III and IV13C-NMR (150MHz, CDCl3) and1H-NMR (600MHz, CDCl3) spectroscopic data
Embodiment 2：Acetylene compound active determination in vitro
External 5-HT, NA and DA monoamine neurotransmitter reuptake has been carried out to each compound of said extracted, separation and has inhibited experiment.
1. experiment material source
1. animal
Male SD rat is purchased from Shanghai Si Laike experimental animal responsibilities Co., Ltd, weight：200-220g, the animal quality certification
Number：SCXK (Shanghai) 2012-0002, feeding room rank：SPF grades of animal house conventinal breedings.
2. reference substance
Fluoxetine hydrochloride is purchased from chemistry division department of Shanghai Institute of Pharmaceutical Industry, and desipramine is purchased from Sigma companies, 6- hydroxyls
Base dopamine is purchased from Sigma companies.
2. prepared by experiment material
1. the preparation of test sample
I. the preparation of compound
Chemical compounds I, II, III and/or IV are dissolved in 10%DMSO (dimethyl sulfoxide (DMSO), a kind of pharmacology common solvent) respectively, match
The solution of 1mg/ml is made, then diluting 10 times again makes a concentration of 100 μ g/ml.
2. the preparation of control drug：
I. Fluoxetine hydrochloride 1.73mg is dissolved in 0.5ml distilled waters, and making a concentration of 0.1mM of its end reaction, (100% inhibits3The reuptake of H-5-HT)
II. desipramine 1.51mg is dissolved in 0.5ml distilled waters, and making a concentration of 0.1mM of its end reaction, (100% inhibits3H-
The reuptake of NA)
III.6- hydroxyl dopamines 1.03mg is dissolved in 0.5ml distilled waters, makes its end reaction concentration：(100% inhibits 0.1mM
3The reuptake of H-DA)
3. the preparation of brain synaptosome
I.5-HT with NA can brain synaptosome preparation
Brain is taken out rapidly after rat broken end, is placed in the physiological saline of precooling (4 DEG C), removes pia mater and blood vessel group
It knits.3g cerebral cortexes are taken, are put into the cold sucrose solution of 30ml 0.32M.It is homogenized and (kept low with supersonic cell pulverizer
Temperature), 4 DEG C of equilibrium centrifugations (1500g, 10min) then take supernatant to centrifuge (20000g, 30min).Supernatant is abandoned or adopted, it is heavy to continue to employ
It forms sediment, i.e. the crude extract of synaptic knob.This sediment fraction again with the cold sucrose solution of 0.32M suspend after be carefully layered on by tube bottom according to
In the secondary cold Sucrose gradient solutions for spreading 1.2M and 0.8M (each 10ml), 4 DEG C of equilibrium centrifugation (38000g) 60min.Use puncture needle
Suspension band at 0.8-1.2M sucrose interfaces of careful collection, is placed in mixing in the cold sucrose solution of 10ml 0.32M, 4 DEG C balance from
The heart (20000g) 30min, sediment are refined brain synaptosome.Sediment is suspended in a small amount of Tris-Krebs buffer solutions
(Tris:15mM；NaCl:94.7mM；KCl:4.7mM；CaCl2:0.5mM；MgSO4:2.4mM；NaH2PO4:1.2mM；
Glucose:10.6mM；Ascorbic:0.57mM；Pargyline:In 0.01mM), suspension is measured with total protein assay kit
In protein content.
The preparation of II.DA energy brain synaptosomes
Brain is taken out rapidly after rat broken end, is placed in the physiological saline of precooling (4 DEG C), removes pia mater and blood vessel group
It knits.The remaining brain tissue that 2g brains is taken to go after cortex, is put into the cold sucrose solution of 20ml 0.32M.With supersonic cell powder
Millstone homogenate (keeps low temperature), and 4 DEG C of equilibrium centrifugations (1500g, 10min) then take supernatant to centrifuge (20000g) 30min.It abandons
Supernatant is taken, continues to employ the crude extract of precipitation, i.e. synaptic knob.Sediment is suspended in a small amount of Tris-Krebs buffer solutions, is used
Total protein assay kit measures the protein content in suspension.
3. the reuptake of monoamine inhibits experiment
1.0ml Tris-Krebs buffer solutions (logical oxygen 15min in advance) are first added in test tube, it is small then to add in 20 μ l cynapses
Then body suspension adds in control sample or 10 μ l of drug to be measured (being operated in 4 DEG C of environment), be uniformly mixed, in 37 DEG C of water-baths
Warm bath 5min.Added in 4 DEG C of environment 10 μ l substrates (3H-5HT、3H-NA or3H-DA；End reaction concentration：300nM), mixing, 37
Warm bath 5min in DEG C water-bath.Then often pipe adds in 3ml and gives cold Tris-Krebs buffer solutions termination reaction, and thin with bull rapidly
Born of the same parents' collector is filtered by glass fiber filter, then rinses test tube and filter 2 times with the same solution of same volume.Remove filter membrane,
60-70 DEG C of drying, filter membrane is put into scintillation vial, adds in toluene scintillation solution, is counted in β-scintillation counter.Blank, 37 DEG C and
Different drug concentrations is 3 multiple pipes.The net intake of synaptosome：37 DEG C of cpm values (active reuptake) subtract 0 DEG C of cpm
It is worth (non-specific aggregation).
The end reaction concentration of positive control drug (Fluoxetine hydrochloride, desipramine, 6- hydroxyls DOPA) is 0.1mM.It is dense herein
Under degree, positive control drug can be with 100% inhibition rat brain synaptosomes to the reuptake of 5-HT, NA and DA.Therefore, with
0.1mM is the end reaction activity of positive control drug, and the inhibiting effect of sample to be tested and corresponding positive control drug are compared
Compared with the screening for carrying out inhibiting effect.
4. experimental result
In order to be prepared for two batches sample respectively in research, tested twice, the CPM values of each group sample are measured, with sun
The inhibition percentage that calculating is compared with the CPM values of drug to be measured, obtains sample of property medicine (positive drug is in terms of 100%).It calculates public
Formula is：
Sample inhibits monoamine neurotransmitter percentage=[1- (CPMSample-CPMControl)/(CPM37℃-CPMControl)] × 100%.
The results are shown in Table 3.
34 samples to be tested of table are to the inhibiting effect of rat brain synaptosomes intake 5-HT, NA, DA
4th, preliminary conclusion：
The reuptake Inhibition test of external monoamine neurotransmitter shows that compound can inhibit the reuptake behavior of NA,
Inhibiting rate is suitable with positive control medicine desipramine；Compound is weaker than same concentrations positive drug salt to the reuptake behavior of 5-HT
Sour Prozac, but also have different degrees of inhibiting effect；Compound does not have inhibiting effect to DA.
Conclusion：
The present invention from radix bupleuri petroleum ether part isolated new compounds i, II, IV and reported compound III.Through body
Outer experiment proves that chemical compounds I, II, III and IV can effectively inhibit the reuptake behavior of NA, and inhibiting rate is with positive control medicine
Xi Paming is suitable；Though and be weaker than positive control drug Fluoxetine hydrochloride to the reuptake behavior of 5-HT, but still have different degrees of suppression
It makes and uses.In conclusion these four compound specific chemical components, belong to low polarity component in plant, and there is specific extraction
Preparation method is expected to exploitation into the specific new antidepressant of effective substance or antidepression pro-drug.
Claims (3)
1. a kind of acetylene compound, which is characterized in that there is following IV structural formula：
2. a kind of method that acetylene compound as described in claim 1 is detached from radix bupleuri, includes the following steps：
(1) prepared by radix bupleuri petroleum ether part：95% ethyl alcohol of radix bupleuri is impregnated, refluxing extraction, extracting solution is concentrated to give medicinal extract, medicinal extract
With petroleum ether extraction, radix bupleuri petroleum ether part is obtained；
(2) crude separation of radix bupleuri petroleum ether part：Radix bupleuri petroleum ether part is added in silicagel column and is detached, with petroleum ether-acetic acid
The mixed liquor elution of ethyl ester different proportion, silica gel thin-layer is qualitative and efficient liquid phase tracer, collects petroleum ether-ethyl acetate ratio respectively
Example is 100:6 parts obtain crude product B；
It uses a dry method on a sample, the mixed liquor of crude product B petroleum ether-ethyl acetate different proportions is eluted, collect petroleum ether-acetic acid
Ethyl ester ratio is 100:1 to 100:2 parts obtain the BA of crude product containing target compound；
(3) it purifies：Crude product BA by silica gel chromatographies column, ODS column chromatographies and SephadexLH-20 gel columns are cleaned respectively, dissolving
In 9:In 1 methanol-chloroform solvent, filter membrane filtration obtains sample BA-1；
(4) preparation of compound：Sample BA-1 is taken, using high performance liquid chromatography, using methanol-water ratio as 87.5:12.5 are
Mobile phase is collected the chromatographic peak that retention time is 22.0-23.9min and is concentrated to dryness, obtains compounds Ⅳ.
3. application of the acetylene compound as described in claim 1 in antidepressant is prepared.
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|---|---|---|---|---|
| WO2003037342A1 (en) * | 2001-10-29 | 2003-05-08 | Ardent Pharmaceuticals, Inc. | Method of treating depression with delta receptor agonist compounds |
| CN103565781A (en) * | 2013-11-06 | 2014-02-12 | 山西大学 | Composite, preparation thereof and application of composite in preparing antidepressant drug |
-
2016
- 2016-02-26 CN CN201610107988.0A patent/CN105646151B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003037342A1 (en) * | 2001-10-29 | 2003-05-08 | Ardent Pharmaceuticals, Inc. | Method of treating depression with delta receptor agonist compounds |
| CN103565781A (en) * | 2013-11-06 | 2014-02-12 | 山西大学 | Composite, preparation thereof and application of composite in preparing antidepressant drug |
Non-Patent Citations (4)
| Title |
|---|
| A qualitative, and quantitative determination and pharmacokineti;MSJiali Liu等;《Journal of Pharmaceutical and Biomedical Analysis》;20150409;第111卷;第257-265页 * |
| Inhibitory effects of oenanthotoxin analogues on GABAergic currents in cultured rat hippocampal neurons depend on the polyacetylenes" polarity;Paulina Wyrembek等;《European Journal of Pharmacology》;20120315;第683卷(第1-3期);第35-42页 * |
| Lignans and Polyacetylenes from Bupleurum acutifolium;Alejandro F. Barrero等;《J. Nat. Prod.》;19990619;第62卷(第7期);第946-948页 * |
| Notiz uber die Inhaltsstoffe von Bupleuuum-Arten;Ferdinand Bohlmann等;《Chem. Ber.》;19711231;第104卷(第6期);第2030-2032页 * |
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