CN102241661B - Preparation method of compound 5'-methoxy-3',4'-methylenedioxy cinnamic acid iso-butyl amide - Google Patents
Preparation method of compound 5'-methoxy-3',4'-methylenedioxy cinnamic acid iso-butyl amide Download PDFInfo
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- 229940125898 compound 5 Drugs 0.000 title abstract description 3
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- 238000000605 extraction Methods 0.000 claims abstract description 109
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- 238000000034 method Methods 0.000 claims description 67
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Abstract
The invention belongs to the field of traditional Chinese medicine pharmacy, and relates to a preparation method of high-purity amide compound 5'-methoxy-3',4'-methylenedioxy cinnamic acid iso-butyl amide by extracting from Piper plants. The preparation method provided by the invention comprises the following steps: (1) extraction: adding medicinal materials into an extraction tank, adding a solvent for extraction, filtering, and concentrating the extraction liquid to obtain an extract; (2) refining: dissolving the extract with a solvent, adding the dissolved extract to a chromatographic column, eluting, collecting the eluate, and concentrating the eluate to be dry to obtain an extract; and (3) separation and purification: dissolving the extract with a solvent, adding the dissolved extract to a chromatographic column, eluting, detecting by thin-layer chromatography, combining the eluate containing 5'-methoxy-3',4'-methylenedioxy cinnamic acid iso-butyl amide, concentrating the combined eluate, recrystallizing, and drying to obtain 5'-methoxy-3',4'-methylenedioxy cinnamic acid iso-butyl amide.
Description
Technical field
The invention belongs to field of traditional Chinese medicine pharmacy, particularly a kind of from Piper plant, extract preparation high purity amides 5 '-methoxyl group-3 ', the method for 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Background technology
Dysthymia disorders is to take a kind of mental disorder that remarkable and lasting emotional handicap is cardinal symptom, take depressed, lose interest, pessimism as main, the symptoms such as, appetite stimulator too low with psychomotor agitation or sluggishness, self-assessment and insomnia.Depression rate is very high, almost in every 7 grownups, just have 1 patients with depression, so it is called as the flu in psychiatry.The < < World Health Report > > in 2002 delivering according to the World Health Organization, dysthymia disorders has become the fourth-largest illness in the world at present, to the year two thousand twenty dysthymia disorders, may become and be only second to cardiopathic second largest disease.The misery that dysthymia disorders causes patient and family members thereof, the loss that society is caused is that other diseases is incomparable.The major cause that causes this situation is that society lacks correct understanding to dysthymia disorders, and prejudice makes patient be reluctant that psychiatric department is medical.In China, instrument has 2% patients with depression to accept treatment, and a large amount of patients can not get diagnosis and treatment timely, and sb.'s illness took a turn for the worse, even occurs the serious consequence of committing suiside.On the other hand, because the common people lack the knowledge of relevant dysthymia disorders, to there is depressive symptom, person thinks it is to be disgruntled by mistake, can not give due understanding and Emotional support, and patient is caused to larger psychological pressure, and the state of an illness is further worsened.
Dysthymia disorders is mainly divided into following several types:
One, endogenous depression has lazyness, slow-witted, change, sorrow, worry " five levy " (brain biogenic amine draw to or definitely not enough).
Two, masked depression Hypoemotivity and melancholy symptom are not obvious, usually show as various body malaise symptoms, as palpitaition, uncomfortable in chest, middle epigastric discomfort, breathe hard, perspire, become thin, insomnia etc.
Three, adolescent depression, can cause student to produce difficulty of learning, and attention laxes, hypomnesis, and achievement declines comprehensively or declines suddenly, is weary of studying, probably learns, plays truant or refuse and learn.
Four, secondary depression if any hyperpietic, take after depressor, cause mood to continue melancholy, downhearted.
Five, post-natal depression its particularly to the baby of oneself produce strong compunction, feel oneself inferior, hate, frightened or detest child's abnormality psychology.The thing of cryying, have a sleepless night, do not feel like eating, melancholy, is the common sympton of this class patients with depression.
Six, white collar dysthymia disorders, the young women disorders of the neuroendocrine system of suffering from dysthymia disorders, normal physiological period is also upset, symptom is varied, except depression, depressed, occupied with nothing, like sulky, worry beyond measure, insomnia, dreaminess, giddy, the main mental symptom such as forgetful, apocleisis, feel sick, the digestion and absorption function diagonosis of disorder such as vomiting, abdominal distension, gynaecology's symptom is quite a few sees for menoxenia, abdominal pain in menstruation etc.
At present, the antidepressant drug of application is mainly the 5 one hydroxy-tryptamine reuptake inhibitors such as fluoxetine, paroxetine and Sertraline clinically, but because depression mechanism is complicated, induced factor is more, medicine for a certain single link is often difficult to obtain satisfactory effect, synthetic thymoleptic exist that antidepressant spectrum is narrow, side effect large, medicine valency is high and stop the defects such as approximately easily recurrence mostly, also may occur that dry, blurred vision, sinus tachycardia, constipation, uroschesis, glaucoma are aggravated, memory dysfunction in clinical use procedure; Block suprarenin a1 acceptor, may occur strengthening hypotensive effect, postural hypotension, giddy, the reflex tachycardia of Prazosin; Cloudy stagnant histamine h1 acceptor, can occur strengthening central depressant effect, calmness, drowsiness, put on weight, reduce blood pressure; , can there is the problems such as extrapyramidal symptoms, endocrine alteration in cloudy stagnant Dopamine HCL d2 acceptor.Therefore, aspect the research and development of thymoleptic, more and more focus on finding activated lead compound both at home and abroad from natural product, by structural modification, find new antidepressant drug.
Chinese patent (application number: 200410025493.0) disclose laetispicine in great Ye Betel and suppressed the application of central nervous system monoamine transmitters reuptake relevant pharmaceutical composition kind in preparation treatment.US Patent No. 6858648 also discloses laetispicine and the application of homologue aspect the mental diseases such as antidepressant thereof.
5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides is a kind of active substance in laetispicine, its structure is:
The inventor is further carrying out great Ye Betel antidepressant basic substance when research, find 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides also has obvious antidepressant activity, and external activity is better than laetispicine.Secondly, 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides is the highest amides of content in great Ye Betel, can infer thus 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides is one of important activity composition of great Ye Betel, therefore in batches prepare 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides as bulk drug by necessary.
In prior art, prepare 5 '-methoxyl group-3 ', the method for 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides adopts organic solvent extraction, silica gel column chromatography is separated repeatedly, often, complicated operation, should not prepare in batches in separation, and the compound purity obtaining is not high, quality unstable is poor.
In order to address the above problem, we constantly study intensively and explore, and have passed through a large amount of experiment screenings, have obtained a kind of technique simple, scientific and reasonable preparation 5 '-methoxyl group-3 ' and, the method for 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Summary of the invention
The object of the invention be to provide a kind of prepare 5 '-methoxyl group-3 ', the method for 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
5 '-methoxyl group-3 that the present invention prepares ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides purity is high, and quality stability is good.
The present invention prepare 5 '-methoxyl group-3 ', the medicinal material of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides is selected: the over-ground part of Piper plant great Ye Betel or P. austrosinense.
Preparation process process following steps of the present invention:
Step 1: extract, use solvent extraction medicinal material, extracting solution, through concentrated, obtains medicinal extract;
Step 2: chromatographic column is refining, and medicinal extract dissolution with solvents, joins in chromatographic column, solvent wash-out, collects elutriant, concentrates to obtain refining extract;
Step 3: chromatographic column separation and purification, after dissolving, refining extract joins in chromatographic column, and solvent wash-out, collects elutriant, thin layer chromatography detects, merge contain 5 '-methoxyl group-3 ', the elutriant of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, concentrated, recrystallization, obtains;
Wherein,
In step 1: medicinal material is the over-ground part of Piper plant great Ye Betel or P. austrosinense.
Solvent is selected from: methyl alcohol, ethanol or water.Preferred alcohol or its aqueous solution.Extracting mode is selected: refluxing extraction, supersound extraction, flash extraction, decocting method, temperature are soaked method, enzyme process assisted extraction, continuous dynamic countercurrent extraction, diacolation extraction, microwave extraction or ultra-high voltage and extracted.Be preferably refluxing extraction.
In step 2: solvent is selected from: methyl alcohol, ethanol or water.Preferred alcohol or its aqueous solution.Chromatographic column is selected: macroporous adsorptive resins, ion exchange resin column.Preferred macroporous adsorbent resin chromatography post, the preferred AB-8 of resin and D101, resin-crude drug ratio is 1: 4-1: 1.Wash-out solvent is selected from: methyl alcohol, ethanol or water; Preferred alcohol or its aqueous solution.
In step 3: chromatographic column is selected according to filler: macroporous adsorptive resins, ion exchange resin column, silica gel, dextrane gel and C18 bonded silica gel.Preferred silica gel, dextrane gel and C18 bonded silica gel chromatographic column.More preferably 200-300 object silica gel, moving phase is selected from: a kind of or more than one mixing in methyl alcohol, ethanol, propyl alcohol, propyl carbinol, ethyl acetate, Virahol, sherwood oil, isopropylcarbinol, water.Preferred petroleum ether-ethyl acetate system.The solvent that recrystallization is selected is: a kind of or more than one mixing in methyl alcohol, ethanol, propyl alcohol, propyl carbinol, ethyl acetate, Virahol, sherwood oil, ethyl acetate, isopropylcarbinol, water.Be preferably ethanol-water system.
The preferred preparation method of the present invention is:
Step 1,
The material of getting it filled, coarse reduction, is placed in extractor, adds the solvent of the 30-95% of 5-20 times of weight, refluxing extraction 1-4 time, each 5 minutes-4 hours, filter, concentrating under reduced pressure becomes medicinal extract;
Step 2,
The dissolution with solvents of 30-95% for medicinal extract, adds in chromatographic column, and with the eluent wash-out of 30-95%, wash-out 1-4BV, collects elutriant, is concentrated into dryly, obtains extract;
Step 3,
Extract is through mixing sample, adding in chromatographic column, is the petroleum ether-ethyl acetate gradient elution of 1-1: 10-1 by volume ratio, and thin-layer method detects, merge contain 5 '-methoxyl group-3 ', the elutriant of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, is concentrated into absence of liquid, separates out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
The preferred preparation method of the present invention is:
Step,
The get it filled over-ground part of material Piper plant great Ye Betel or P. austrosinense, coarse reduction, be placed in extractor, add the solvent of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filter, the solvent of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filters, merge filtrate twice, 65 ℃ of concentrating under reduced pressure become the medicinal extract of RD1.3-1.35;
Step 2,
The dissolution with solvents of 30-60% for medicinal extract, adds in the chromatographic column of D101 macroporous adsorbent resin, and with the eluent wash-out of 30-60%, elutriant discards, and uses 70-95% eluent wash-out instead, collects elutriant, is concentrated into dryly, obtains extract;
Step 3,
Extract is through mixing sample, adding in the chromatographic column that is filled with silica gel, is 5: 1 and 3: 1 petroleum ether-ethyl acetate gradient elutions by volume ratio respectively, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through eluent washing, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
The most preferred preparation method of the present invention is:
Step 1,
The get it filled over-ground part of material Piper plant great Ye Betel or P. austrosinense, coarse reduction, be placed in extractor, add the ethanol of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filter, the ethanol of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filters, merge filtrate twice, 65 ℃ of concentrating under reduced pressure become the medicinal extract of RD1.3-1.35;
Step 2,
The dissolve with ethanol of 30-60% for medicinal extract, adds in the chromatographic column of D101 macroporous adsorbent resin, and with the ethanol elution of 30-60%, elutriant discards, and uses 70-95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains extract;
Step 3,
Extract, through mixing sample, add in the chromatographic column that is filled with silica gel, is 5: 1 and 3: 1 petroleum ether-ethyl acetate gradient elutions by volume ratio respectively, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaims solvent, is concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior first two dioxy base TRANSCINNAMIC ACID isobutyl-acid amides, altogether 1.68g.
The most preferred preparation method's of the present invention specific examples in an embodiment.
The compound preparing by preparation method of the present invention through LC-MS,
1hNMR,
13cNMR and DEPT spectrum is measured, in conjunction with data in literature, can prove 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
5 '-methoxyl group-3 that the present invention prepares ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides has good effect for treatment mental disorder.
Preparation method of the present invention is that the screening through science gets, compared with prior art, only through twice chromatographic, preparation technology simplifies greatly, safe preparation process, and materials cost is low, be applicable to scale operation, 5 '-methoxyl group-3 that obtain after preparation ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides purity is high, and quality stability is good.
Accompanying drawing explanation
Fig. 1 great Ye Betel extractive HPLC color atlas
Figure 25 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides product HPLC color atlas
The product hydrogen spectrum that Fig. 3 the present invention prepares
The product carbon spectrum that Fig. 4 the present invention prepares
Embodiment
By following specific embodiment, the invention will be further described, but not as limitation of the present invention.
Embodiment 1, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, concentrated as for, obtain 33g great Ye Betel extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization (alcohol concn is 95%), obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, altogether 1.68g.(seeing accompanying drawing 1-4)
Embodiment 2, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains 21g P. austrosinense extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 400g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, altogether 0.85g.
Embodiment 3: preparation method of the present invention
Substantially the same manner as Example 1, difference is that extracting method adopts 10 times of amount 95% ethanol ultrasonic extraction twice, each 30 minutes, finally obtain 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides 1.35g.
Embodiment 4: preparation method of the present invention
Substantially the same manner as Example 1, difference is that extracting method adopts the flash extraction twice of 10 times of amount 95% ethanol, each 5 minutes, finally obtain 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides 1.8g.
Embodiment 5, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 70% ethanol of 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 6, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 70% ethanol of 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 60% dissolve with ethanol, uses 60% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 7, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 70% ethanol of 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 30% dissolve with ethanol, uses 30% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 8, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 70% ethanol of 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 30% dissolve with ethanol, uses 30% ethanol elution, and elutriant discards, and uses 70% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 9, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 150kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 150kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 10, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 80% ethanol of 100kg, refluxing extraction 2 hours, filters, 80% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 40% dissolve with ethanol, uses 40% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 11, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 80% ethanol of 100kg, refluxing extraction 1 hour, filters, 80% alcohol reflux of dregs of a decoction continuation use 100kg 3 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 40% dissolve with ethanol, uses 40% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 12, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 0.5 hour, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 3 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 13, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 1.5 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 1.5 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 14, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 80% ethanol of 100kg, refluxing extraction 2 hours, filters, 80% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgAB-8 macroporous adsorbent resin to 2L with 40% dissolve with ethanol, uses 40% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 15, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgAB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 16, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 1 hour, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 3 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgAB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 17, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% methyl alcohol of 100kg, refluxing extraction 2 hours, filters, and the dregs of a decoction continue to extract 2 hours with 95% methanol eddy of 100kg, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgAB-8 macroporous adsorbent resin to 2L with 50% dissolve with methanol, and with 50% interior alcohol wash-out, elutriant discards, and uses 95% methanol-eluted fractions instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 18, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds and is filled with in ion exchange resin column to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 19, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% methyl alcohol of 100kg, refluxing extraction 2 hours, filters, and the dregs of a decoction continue to extract 2 hours with 95% methanol eddy of 100kg, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds and is filled with in ion exchange resin column to 2L with 50% dissolve with methanol, uses 50% methanol-eluted fractions, and elutriant discards, and uses 95% methanol-eluted fractions instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 20, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract adds in the chromatographic column that is filled with 600g glycan gel, use 60% methanol-eluted fractions, thin-layer method detects, and merges the single-point composition of sample size maximum, reclaims solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 21, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600gC18 bonded silica gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 22, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgAB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600g glycan gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 23, preparation method of the present invention
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kg AB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600gC18 bonded silica gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 24, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 70% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 25, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 70% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 60% dissolve with ethanol, uses 60% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 26, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 70% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 30% dissolve with ethanol, uses 30% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 27, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 70% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 70% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 30% dissolve with ethanol, uses 30% ethanol elution, and elutriant discards, and uses 70% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 28, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 150kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 150kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃).
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract.
Extract is through mixing sample, add in the chromatographic column that is filled with 600g 200-300 order silica gel, with petroleum ether-ethyl acetate (5: 1) and (3: 1) gradient elution, thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle.Yellow particle becomes white particle through petroleum ether, then through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 29, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 80% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 80% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 40% dissolve with ethanol, uses 40% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 30, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 80% ethanol that adds 100kg, refluxing extraction 1 hour, filters, 80% alcohol reflux of dregs of a decoction continuation use 100kg 3 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 40% dissolve with ethanol, uses 40% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 31, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 0.5 hour, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 3 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 32, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 1.5 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 1.5 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 33, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 80% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 80% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD AB-8 macroporous adsorbent resin to 2L with 40% dissolve with ethanol, uses 40% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 34, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD AB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 35, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 1 hour, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 3 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgAB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 36, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% methyl alcohol that adds 100kg, refluxing extraction 2 hours, filters, the dregs of a decoction continue to extract 2 hours with 95% methanol eddy of 100kg, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD AB-8 macroporous adsorbent resin to 2L with 50% dissolve with methanol, and with 50% propyl alcohol wash-out, elutriant discards, and uses 95% methanol-eluted fractions instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate wash-out of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 37, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds and is filled with in ion exchange resin column to 2L with 50% dissolve with ethanol, uses 50% ethanol elution, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 38, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% methyl alcohol that adds 100kg, refluxing extraction 2 hours, filters, the dregs of a decoction continue to extract 2 hours with 95% methanol eddy of 100kg, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds and is filled with in ion exchange resin column to 2L with 50% dissolve with methanol, uses 50% methanol-eluted fractions, and elutriant discards, and uses 95% methanol-eluted fractions instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g 200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5: 1 and 3: 1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3: 1 systems, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 39, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600g glycan gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 40, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600gC18 bonded silica gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 41, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kg AB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600g glycan gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 42, preparation method of the present invention
Get P. austrosinense over-ground part 10kg, coarse reduction, put in pilot extraction tank, 95% ethanol that adds 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, be evaporated to the medicinal extract of RD1.3-1.35 (65 ℃);
Medicinal extract, adds in the chromatographic column that is filled with 3kg AB-8 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, uses 50% wash-out, and elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, add in the chromatographic column that is filled with 600gC18 bonded silica gel, use 60% methanol-eluted fractions, thin-layer method detects, the single-point composition that merges sample size maximum, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 43, preparation method of the present invention
Substantially the same manner as Example 2, difference is that extracting method adopts 10 times of amount 95% ethanol ultrasonic extraction twice, each 30 minutes, finally obtain 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Embodiment 44, preparation method of the present invention
Substantially the same manner as Example 2, difference is that extracting method adopts the flash extraction twice of 10 times of amount 95% ethanol, each 5 minutes, finally obtain 5 '-methoxyl group-3 ', 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
Claims (6)
1.5 '-methoxyl group-3 ', the preparation method of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, is characterized in that, process following steps:
Step 1,
The material of getting it filled, coarse reduction, is placed in extractor, adds the ethanol of the 30-95% of 5-20 times of weight, refluxing extraction 1-4 time, each 5 minutes-4 hours, filter, concentrating under reduced pressure becomes medicinal extract;
Step 2,
The dissolve with ethanol of 30-95% for medicinal extract, adds in chromatographic column, and with the ethanol elution of 30-95%, wash-out 1-4BV, collects elutriant, is concentrated into dryly, obtains extract;
Step 3,
Extract is through mixing sample, add in chromatographic column, the petroleum ether-ethyl acetate gradient elution that is 1-1:10-1 by volume ratio, thin-layer method detects, merge contain 5 '-methoxyl group-3 ', the elutriant of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, is concentrated into absence of liquid, separates out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides;
Wherein,
In step 1: medicinal material is the over-ground part of Piper plant great Ye Betel or P. austrosinense;
In step 2: chromatographic column is selected macroporous adsorbent resin D101, resin-crude drug is than being 1:4-1:1;
In step 3: chromatographic column filler is selected 200-300 object silica gel, and moving phase is selected from: petroleum ether-ethyl acetate system, the solvent that recrystallization is selected is ethanol-water system.
2. preparation method according to claim 1, is characterized in that, process following steps:
Step 1,
The get it filled over-ground part of material Piper plant great Ye Betel or P. austrosinense, coarse reduction, be placed in extractor, add the ethanol of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filter, the ethanol of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filters, merge filtrate twice, 65 ℃ of concentrating under reduced pressure become the medicinal extract of RD1.3-1.35;
Step 2,
The dissolve with ethanol of 30-60% for medicinal extract, adds in the chromatographic column of D101 macroporous adsorbent resin, and with the ethanol elution of 30-60%, elutriant discards, and uses 70-95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains extract;
Step 3,
Extract is through mixing sample, adding in the chromatographic column that is filled with silica gel, is 5:1 and 3:1 petroleum ether-ethyl acetate gradient elution by volume ratio respectively, and thin-layer method detects, the single-point composition that merges sample size maximum under 3:1 system, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
3. preparation method according to claim 1, is characterized in that, process following steps:
Step 1,
The get it filled over-ground part of material Piper plant great Ye Betel or P. austrosinense, coarse reduction, be placed in extractor, add the ethanol of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filter, the ethanol of the 70-95% of 10-15 times of weight, refluxing extraction 2 hours, filters, merge filtrate twice, 65 ℃ of concentrating under reduced pressure become the medicinal extract of RD1.3-1.35;
Step 2,
The dissolve with ethanol of 30-60% for medicinal extract, adds in the chromatographic column of D101 macroporous adsorbent resin, and with the ethanol elution of 30-60%, elutriant discards, and uses 70-95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains extract;
Step 3,
Extract is through mixing sample, adding in the chromatographic column that is filled with silica gel, is 5:1 and 3:1 petroleum ether-ethyl acetate gradient elution by volume ratio respectively, and thin-layer method detects, the single-point composition that merges sample size maximum under 3:1 system, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through 95% ethyl alcohol recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
4. preparation method according to claim 1, is characterized in that, described preparation method is:
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, 65 ℃ of medicinal extract that are evaporated to RD1.3-1.35;
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, and with 50% ethanol elution, elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5:1 and 3:1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3:1 system, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
5. preparation method according to claim 1, is characterized in that, described preparation method is:
Get P. austrosinense over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds 95% ethanol of 100kg, refluxing extraction 2 hours, filters, 95% alcohol reflux of dregs of a decoction continuation use 100kg 2 hours, filter, merge twice filtrate, 65 ℃ of medicinal extract that are evaporated to RD1.3-1.35;
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, and with 50% ethanol elution, elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains P. austrosinense extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 400g200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5:1 and 3:1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3:1 system, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
6.5 '-methoxyl group-3 ', the preparation method of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides, is characterized in that, process following steps:
Get great Ye Betel over-ground part 10kg, coarse reduction, puts in pilot extraction tank, adds the flash extraction twice of 95% ethanol of 100kg, and each 5 minutes, filter, merge twice filtrate, 65 ℃ of medicinal extract that are evaporated to RD1.3-1.35;
Medicinal extract, adds in the chromatographic column that is filled with 3kgD101 macroporous adsorbent resin to 2L with 50% dissolve with ethanol, and with 50% ethanol elution, elutriant discards, and uses 95% ethanol elution instead, collects elutriant, is concentrated into dryly, obtains 33g great Ye Betel extract;
Extract is through mixing sample, adding in the chromatographic column that is filled with 600g200-300 order silica gel, is the petroleum ether-ethyl acetate gradient elution of 5:1 and 3:1 by volume ratio, and thin-layer method detects, the single-point composition that merges sample size maximum under 3:1 system, reclaim solvent, be concentrated into absence of liquid, separate out yellow particle, yellow particle becomes white particle through petroleum ether, again through ethanol-water system recrystallization, obtain 5 '-methoxyl group-3 ', the white crystals of 4 '-inferior methylenedioxy group TRANSCINNAMIC ACID isobutyl-acid amides.
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| CN113209165A (en) * | 2017-11-01 | 2021-08-06 | 苏州颐华生物医药技术股份有限公司 | Piper laetispicum extract and preparation method and application thereof |
| CN109053403A (en) * | 2018-06-04 | 2018-12-21 | 上海博桂文化传播有限公司 | A method of extracting separation 2- methoxyl group -1,4- naphthoquinones from CAULIS IMPATIENTIS |
| CN112079692B (en) * | 2020-10-29 | 2023-05-12 | 中国热带农业科学院环境与植物保护研究所 | Compound for preventing and treating plant pathogenic bacteria and application thereof |
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Non-Patent Citations (2)
| Title |
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| 刘红宇等.华南胡椒化学成分的研究.《天然产物研究与开发》.1995,第7卷(第2期),第20-23页. |
| 华南胡椒化学成分的研究;刘红宇等;《天然产物研究与开发》;19950630;第7卷(第2期);第20-23页 * |
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