CN1292750C - Total alkaloid composition from plant and its pharmaceutical preparation - Google Patents
Total alkaloid composition from plant and its pharmaceutical preparation Download PDFInfo
- Publication number
- CN1292750C CN1292750C CNB03123772XA CN03123772A CN1292750C CN 1292750 C CN1292750 C CN 1292750C CN B03123772X A CNB03123772X A CN B03123772XA CN 03123772 A CN03123772 A CN 03123772A CN 1292750 C CN1292750 C CN 1292750C
- Authority
- CN
- China
- Prior art keywords
- total alkaloids
- precipitation
- solution
- preparation
- add
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention discloses a total alkaloid composition having anticancer activities, which is derived from plants, an extraction method of the total alkaloid composition, an application of the total alkaloid composition, a medical preparation comprising the composition, and a preparation method of the medical preparation. The plants are taken from pink plumepoppy herb, greater celandine herb, eomecon chionantha hance or asiatic toddalia root. The preparation has the functions of strongly inhibiting MRP gene expression, and favorably inhibiting MDRI expression induced by Ara-C, and has favorable antitumoral effect.
Description
Technical field that the present invention belongs to
The present invention relates to have active anticancer plant origin total alkaloids compositions and extracting method and purposes, comprise pharmaceutical preparation of said composition and preparation method thereof, described plant is selected from Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance or Radix Toddaliae Asiaticae.
Background technology
Alkaloid is one of main effective medicinal components in many plants, also is one of important content of present Chinese medicine research.
Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae are the plant amedica that still excludes the Pharmacopoeia of the People's Republic of China.Wherein Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance all belong to Papaveraceae, formal name used at school is respectively Macleayacordata (Willd.) R.Br., Chelidonium majus L., Eomecon chionantha Hance, Radix Toddaliae Asiaticae is root, the leaf of Rutaceae Radix Toddaliae Asiaticae platymiscium Radix Toddaliae Asiaticae, formal name used at school is Toddaliaasiatica (L.) Lam. nature and flavor and effect: wherein Herba Macleayae Cordatae, Herba Chelidonii, the equal bitter in the mouth of Eomecon chionantha Hance, and cold in nature, poisonous.Have detumescence, detoxifcation, insecticidal function, medicine is used for the furunculosis abscess, malignant boil ulcer, and scald, stubborn dermatitis etc. are external, do not take orally.Radix Toddaliae Asiaticae acrid in the mouth, little hardship, temperature, poisonous; Function cures mainly: expelling wind and removing dampness, subduing swelling and detoxicating; Be used for traumatic injury, rheumatic arthritis, intercostal neuralgia is used for set a broken bone, venom outward.Among the peoplely many mash external application or fry in shallow oil water smoking and wash with fresh fruit, root or herb.
Extensive utilization to Herba Macleayae Cordatae is used as biological pesticide and biological double-effect manure when Yuexi County, first-elected Anhui Province.Chinese patent CN1256873, CN1351835 are the invention of Herba Macleayae Cordatae with used as pesticides.The Chinese invention patent that Herba Macleayae Cordatae is used for medicine has CN1199640, and the Chinese medicine liquid for eliminating acne and skin care is to make topical agent by Herba Macleayae Cordatae root bark and Herba Bidentis Bipinnatae after Chinese liquor or soak with ethanol; CN1060215, a kind of preparation method for the treatment of the liver antidote will go heart Semen Nelumbinis, Rhizoma Phyllostachydis henonis and root of Herba Macleayae Cordatae three flavor Chinese medicines to dry or dry, and grind into powder mixes YUGAN spirit powder with brown sugar.Obviously, these two patents still belong to more primary degree to the utilization of Herba Macleayae Cordatae.The Chinese patent that relates to Herba Chelidonii has one kind of CN1401368 to treat the Chinese medicine preparation Herba Chelidonii extracting solution of diabetes, and this medicinal liquid extracting method of this patent is simple, though through once using pure deproteinization operation, very coarse, so only be suitable for external.Relate to Herba Chelidonii and also have 20 of other patents, be compound preparation.The Chinese patent that relates to Radix Toddaliae Asiaticae has the exterior-applied medical wine of CN1342485 treatment wound, the CN1237428 medicated wine for curing traumatic injury, CN1173364 medicated wine and preparation method thereof (treatment traumatic injury), CN1156052 treats the sick medicine of toothache for one kind, these patent drugs are compound recipe, and medicine for external use is formed and belonged to mostly in maximum reaching by 37 flavor Chinese medicines.The Chinese patent that relates to Eomecon chionantha Hance has the CN1378853 charcoal powder capsule, is to be made up of 12 flavor medical materials; Its preparation technology is cleaning, immersion, decoction, filtration, concentrated, spraying, centrifugal, dry, incapsulates to form.Belong to a kind of obviously and extract more extensively, can't determine effective ingredient.Chinese patent CN1255061 chelerythrine and radiation method therapeutic alliance tumor, this patent is applied for by U.S. Azithromycin Development Co., Ltd.Chelerythrine is one of main component in the alkaloids of plants such as Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae, this patent to chelerythrine use which kind of dosage form not with explanation.Through retrieve FDA pharmacy resource information with active component, also can be found without any record, can think that therefore total alkaloids preparations such as all being difficult Herba Macleayae Cordatae that chelerythrine is a main component, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae at present both at home and abroad is in clinical practice.
Summary of the invention
Total alkaloids of the present invention or its pharmaceutical preparation (following abbreviate as sometimes " total alkaloids " or " total alkali " or " total alkaloids preparation " or " total alkali preparation ") are characterised in that the total alkaloids that utilizes natural plants Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae extract, through removing its invalid components, reduce the big composition of its side effect, keep a series of processing such as its effective ingredient of purification and form the salt of total alkaloids, consist predominantly of the salt of chelerythrine and the salt of Sanguinarine.
Its dosage form is a solid preparation, comprises tablet, slow releasing tablet, capsule, slow releasing capsule, soft capsule and pellet etc., and its dosage form can also be injection, injectable powder, infusion solution and fat milk injection.
As everyone knows, contain multiple composition in Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, the Radix Toddaliae Asiaticae total alkali, based on Sanguinarine (sanguinarine) and chelerythrine (chelerythrine), other also has protopine, allocryptopine etc.According to existing document announcement, in Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, the toddalia asiatica total alkali extractive, Sanguinarine and chelerythrine account for the 60%-80% of total alkali content.In the adverse reaction of tcm record, it is medicinal directly to utilize Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae herb or root bark to do, and causes cardiac toxicity when heavy dose of easily.
" emergency medicine " 1999 the 8th volume fourth phase has been reported " Herba Macleayae Cordatae poisoning induced malignant arrhythmia 1 example ", and poisoning symptom just appears in gram surplus decoction root of Herba Macleayae Cordatae 100 of patient after 10 minutes.And Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae total alkali main component be studies show that Sanguinarine can be used for treating for skin disease (Merch mdex seventh Edition, 1960; P918) studies show that further that Sanguinarine has the blocking oxide phosphorylation and suppresses the effect of reverse transcription.Simultaneously, Sanguinarine also demonstrates makes no exception to normal cell and cancerous cell, does not have selectivity; Its powerful cytotoxicity shows rapid and intensive glutathione depletion action; (" Cancer chemother Pharmacal " 2003, APr, 17).In addition, also there are some researches show.Sanguinarine and angiotensin I (ATI) acceptor interaction be with a kind of slowly, reversible and non-competing hardly mode carries out (" Eur.J.Pharmatol " 2003, Jan, 5; 458 (3): 257-262).Therefore, in total alkaloids of the present invention and pharmaceutical preparation thereof, just need to remove the invalid components in the total alkali, reduce the big composition of toxicity, active constituent-enriched, should make full use of the synergism of each composition simultaneously, with the effect of performance total alkaloids in anticancer.
In total alkaloids of the present invention and the pharmaceutical preparation thereof, the chelerythrine content range is 30%-100% (in the amount of total alkali), Sanguinarine content range 0%-40%, and protopines etc. and impurity content are 0%-30%.
Total alkaloidss such as Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance, Radix Toddaliae Asiaticae can form salt with multiple mineral acid and organic acid, therefore in total alkaloids of the present invention and the pharmaceutical preparation thereof, total alkaloids exists with the form of salt, and the salt of total alkaloids can be inorganic acid salt, also can be acylate.Example hydrochloric acid salt, sulfate, nitrate, citrate, acetate etc.Total alkaloids is used for preparation with the form of salt, helps the preparation convenient drug administration and guarantees the stable of preparation.
The invention discloses the preparation method of total alkaloids and pharmaceutical preparation thereof, comprise the preparation of extraction, purification and the preparation of total alkaloids.
The total alkaloids extracting method is to obtain extract by acid solution immersion, filter pressing, regulates by pH value and removes most of invalid components and the big composition of toxic and side effects, and active constituent-enriched, concrete steps are as follows:
A) Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance or Radix Toddaliae Asiaticae plant amedica are cut into the 0.5-2cm segment, add 4-8 and doubly measure solution, regulate soak PH to 2.0-3.0, stir frequently, soak 16-24h, filter pressing obtains extract.
B) filtering residue adds the acid solution immersion that 4-6 doubly measures PH 2.0-3.0 again, stir frequently, and soaked overnight, filter pressing obtains extract.
C) merge twice extract, regulating extract PH is 6.0-7.0, and standing over night makes the alkaloid precipitation.
D) filter, collect filtrate.
E) regulating filtrate PH is 9.0-9.5, and adds saline solution, makes that the concentration of salt reaches 3%-4% in the filtrate, and standing over night makes the alkaloid precipitation.
F) filter collecting precipitation, get the precipitate of total alkaloids.Yield is 1.5%-2%, and wherein the chelerythrine alkali content is 40%-60% (in the amount of total alkali, down together), and Sanguinarine content is 20%-30%, and other composition is 20%-30%.
Total alkaloids adsorbs through ion exchange resin, and solvent extraction method is refining, further removes invalid components, and is active constituent-enriched.May further comprise the steps:
A. the total alkaloids precipitation is washed to neutrality, adding PH is in the aqueous solution of 2.0-3.0, makes fully dissolving of precipitation.
B. solution is passed through cation exchange resin, be washed to neutrality, reuse ammonia-alcoholic solution eluting is collected eluent.
C. add equal amounts of chloroform in eluent, regulate PH to 9.0-9.5, thermal agitation 15min leaves standstill phase-splitting, tells ammonia-ethanol phase, adds the equal amounts of chloroform extraction more once, merges chloroform extraction liquid twice.
D. under 40-70 ℃, the reclaim under reduced pressure chloroform is to original volume 1/2-1/4.
E. in the chloroform concentrated extract, add the equivalent acidic aqueous solution, transfer solution PH 2.0-3.0, extract 2 times, merge water.
F. water transfer phase PH is 9.0-9.5, and standing over night makes the alkaloid precipitation, filters collecting precipitation.
G. precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, make fully dissolving of precipitation, add appropriate hydrochloric acid while hot, transfer solution PH 4.0-5.0, standing over night makes the alkaloid crystallization, collects crystallization, and is drying to obtain under 40 ℃.
The plant amedica that is used for extract total alkaloids is Herba Macleayae Cordatae herb, Herba Chelidonii herb, Eomecon chionantha Hance herb, the root of Radix Toddaliae Asiaticae, leaf.
The solvent that is used for total alkaloids extracting, extraction is one or both a mixed solvent of water, methanol, ethanol, chloroform, ammonia.
The salt that is used for regulating the total alkaloids solution ion concentration is one or both mixing salt solution of sodium chloride, zinc chloride.
The acid that is used for total alkaloids extracting and refining purification process adjusting pH can be mineral acid, and example hydrochloric acid, sulphuric acid, nitric acid also can be organic acid, as citric acid, acetic acid.
Being used for total alkaloids extracting and refining purification process, to regulate the alkali of pH can be a kind of in ammonia, sodium hydroxide, sodium carbonate, potassium hydroxide, the potassium carbonate or two kinds mixed solution wherein.
Plant total alkaloid of the present invention and pharmaceutical preparation thereof are made up of total alkali salt, pharmaceutic adjuvant and water such as Herba Macleayae Cordatae, and wherein preparation comprises tablet, capsule, soft capsule, slow releasing tablet, slow releasing capsule, pellet, injection, injectable powder (comprising lyophilized injectable powder), infusion solution and fat milk injection.
The pharmaceutical preparation of preparation total alkaloids is all carried out in the workshop more than 100 grades.
The total alkaloids preparation can be all preparation formulations pharmaceutically, mainly comprises solid preparation and ejection preparation two big classes.
The total alkaloids solid preparation mainly contains tablet, capsule, soft capsule, slow releasing tablet, slow releasing capsule, pellet etc.Its preparation method is that total alkaloid salt and pharmaceutic adjuvant are comprised mix homogeneously such as excipient, filler, disintegrating agent, coating materials, slow releasing agent, cosolvent, dispersant, antiseptic, by tabletting/pelletize, existing pharmaceutical equipment such as dress (fillings) capsule etc. and the realization of conventional pharmaceutical technology.
The solid constituent of total alkaloids preparation injection is made up of total alkaloid salt and pharmaceutic adjuvant, and wherein total alkaloid content is 30%-99.8%, and surplus is an adjuvant.
The total alkaloids injection mainly contains injection, injectable powder (comprising lyophilized injectable powder), infusion solution and fat milk injection.Its preparation method is different according to different dosage forms.
Total alkaloid salt is dissolved with water for injection, after the injection of adding metering dissolved with the water solublity pharmaceutic adjuvant, regulator solution PH to 4.0-5.0 was after the microporous filter membrane ultrafiltration, add the injection water and add to ormal weight, carry out packing or carry out dried according to the dosage form needs again.
Preparation total alkaloids injection, the interior total alkaloid contents (in chelerythrine) of injection is 50mg/ml or 100mg/ml before the packing, be sub-packed in the ampoule bottle, every 2ml, sealing by fusing, after the assay was approved, packing is promptly.
Preparation total alkaloids infusion solution, total alkaloid contents (in chelerythrine) is 1mg/ml in the preceding injection of packing, is packed as 100ml, 150ml, 200ml in corresponding infusion bottle, rolls lid, after the assay was approved, packing is promptly.
Preparation total alkaloids injectable powder, after the salt of purified total alkaloids doubly measured the water for injection dissolving with 5-10, after the injection of adding metering fully dissolves with the water solublity pharmaceutic adjuvant, transfer PH to 4.0-5.0, through ultrafiltration, in 40 ℃ of following vacuum dryings, aseptic subpackaged in the 5ml cillin bottle, every bottle contains total alkaloids 100mg or 200mg (in chelerythrine), rolls lid, after the assay was approved, packing promptly.
Preparation total alkaloids lyophilized injectable powder, with the salt of the total alkaloids after refining, doubly measure the dissolving of (in chelerythrine) water for injection with 5-10 after, the injection water solublity pharmaceutic adjuvant that adds metering, transfer PH to 4.0-5.0,, be diluted to every milliliter with water for injection and contain total alkali (in chelerythrine) 100mg or 200mg through ultrafiltration, be sub-packed in the ampoule, every 1ml, after lyophilization, sealing by fusing, after the assay was approved, packing promptly.
Preparation total alkaloids lipomul is that total alkali salt is formed total alkali by pH regulator, is dissolved in the fat solvent, adds water for injection, emulsifying agent and pharmaceutic adjuvant and prepares.
In the injection of total alkaloids preparation, described injection comprises cosolvents such as propylene glycol, glycerol with the water solublity pharmaceutic adjuvant, antioxidant such as sodium sulfite, sodium pyrosulfite, pH buffer agent such as organic principle such as glucose, lactose, mannitol, low molecular dextran, aminoacid and phosphate
Research both at home and abroad shows that all one of main component in total alkaloids chelerythrine is Protein kinase C (PKC) inhibitor.
According to us the in vitro tests that the total alkaloids preparation carries out is shown that this preparation has good inhibition effect to PKC.
Test 1
Existing studies show that.PKC has activation to multiple drug resistance protein (MRP) expression of gene, and pkc inhibitor is expressed the performance inhibitory action by suppressing the PKC activity to mrp gene.
With of the effect of the various pkc inhibitors of H-9 cell analysis to mrp gene.The result shows that when without any pkc inhibitor, the expression of mrp gene is normal in the cell.And educate altogether with following inhibitor, mrp gene is expressed by strong inhibition.
Table 1: the inhibitory action that kinases inhibitor is expressed mrp gene
| Pkc inhibitor | Concentration (/ml) | Test result |
| staurosporine | 100nM | ++++ |
| Total alkaloids preparation of the present invention | 1-5muM | ++++ |
| Bykomycin | 10mM | ++++ |
| Methyl-1，5-dihydroxycinnamate | 32muM | ++++ |
++ ++=suppress mrp gene fully to express
Test 2
There are some researches show, in chemotherapy, cancerous cell is easy to generate the multiple drug resistance under induced by chemotherapeutic agents, H-9 cell and Ara-C educate altogether, make the H-9 cell produce the multiple drug resistance, and this resistance and mrp gene expression have dependency, and H-9 cell and Ara-C (25muM/ml) are educated 10h altogether, can induce MDR1 to express.
Test is incubated following pkc inhibitor Ara-C (25muM/ml) altogether with the H-9 cell, the expression of test MDR1.
Table 2: kinases inhibitor suppresses the expressional function that Ara-c induces MDR1
| Pkc inhibitor | Concentration (/ml) | Test result |
| staurosporine | 30nM | +++ |
| Total alkaloids preparation of the present invention | 5muM | +++ |
| Bykomycin | 10muM | +++ |
| Methyl-1，5-dihydroxycinna mate | 32muM | +++ |
| genisterine | 150muM | + |
| D，L-threo-sphingosine | 5muM | - |
++ ++=suppress the inducing action of Ara-c fully
The inducing action of the inhibition Ara-c of +=weak
-=not suppresses the inducing action of Ara-c
Sanguinarine is a kind of good red pigments, and has the excellent antibiotic effect, and domestic main have a lot of researchs that Sanguinarine is used for topical agent and sterilizing oral preparation abroad as food color, rarely foundly is used for oral and injection.
According to our studies show that, Sanguinarine content in total alkaloids is higher, and microorganism is had stronger inhibitory action, and cancerous cell is also had the good restraining effect, educating altogether with prostate gland cancer cell and Sanguinarine, the result shows that Sanguinarine is 0.9-3.3mM/ml to the fatal dose of cancerous cell.Undoubtedly, Sanguinarine is a kind of good cytotoxic factor.Studies show that further Sanguinarine also has similar toxicity to normal cell, that is to say its poor selectivity, this is to have limited one of principal element that it uses on medicine.
Therefore in a kind of anticancer total alkaloids preparation of development, need to reduce the content of Sanguinarine.Consider that simultaneously Sanguinarine do not find the report of its toxic and side effects as the food color extensive use, also consider the antibiotic and anti-cancer function of Sanguinarine, suitably keep a small amount of Sanguinarine,, can bring into play the antitumaous effect of total alkaloids better with the chelerythrine synergism.Test shows, keeps a spot of Sanguinarine and does not see that under test dose obvious toxic and side effects is arranged.
Pharmacodynamic study shows: under the total alkaloids preparation injection low dosage (mice 60mg/kg, rat 40mg/kg), obviously suppress mice auricle swelling due to the dimethylbenzene and the rat paw edema due to the Ovum Gallus domesticus album, suppression ratio reaches more than 40%.In vitro tests shows that total alkaloids preparation 1-5muM/ml is dose-dependence to the inhibitory action of cancerous cell.
Total alkaloids preparation of the present invention is used for anticancer research, has shown good anti-cancer activity.With preparation of the present invention respectively with human liver cell tumor HepG
2, HL-60 cell, COLO 250 knots, rectum cancer cell educate altogether, find not change Bcl-2 and Bax albumen after the medication, but can cause the proteic significant change of P53 and C-myc, by changing multiple oncogene protein and apoptosis-induced proteic expression, cancer cell specific induction of apoptosis.
Studies show that further total alkali preparation of the present invention suppresses enzymatic activitys such as PKC, PKA, CPK, thus the anticancer increment; Find also that simultaneously total alkali preparation of the present invention has obvious inhibitory action to the angiogenesis of Human umbilical vein endothelial cells, can also cause preformed angiorrhexis; Research and analyse and think, it may suppress angiogenesis by regulating MMP activities.
Toxicologic study shows that the total alkaloids preparation is respectively the acute toxicity of mice and rat: to the LD of mice iv
50With 95% credible 178mg (the 160-198mg/kg)/kg that is limited to; LD to rat sc
50With the 95% credible 251mg/kg (224-281mg/kg) that is limited to.
The total alkaloids preparation shows the Canis familiaris L. long term toxicity test, with 30 times of the daily dosage of clinical plan that is equivalent to respectively to be grown up, 15 times, three kinds of dosage 120mg/kg of 6 times total alkaloids preparation, 60mg/kg, 24mg/kg intramuscular injection respectively, continuous 13 weeks, observe the activity of Canis familiaris L., appetite, defecation, by hair, body weight and breathing, the variation of vital signs such as heart rate, carry out routine blood test, routine urinalysis, liver function, biochemical analysis such as renal function, electrocardiogram, organ coefficient and pathological examination result show: high dose group part Canis familiaris L. decreased heart rate occurs after 13 weeks of administration, the P-R interval, prolong, the basic recovery normally after the drug withdrawal, all the other groups show no obvious abnormalities.Mild edema between the visible cardiac muscle of high dose group part Canis familiaris L., blood vessel is slightly expanded, outside the hyperemia, all the other no abnormality seens.Each organizes the Canis familiaris L. intramuscular injection site a small amount of inflammatory cell infiltration, but does not see obviously hemorrhage, downright bad, cicatrix etc.
The rat long term toxicity test shows, with be equivalent to be grown up 27.5 times, 17.5 times, 10 times three kinds of dosage (110mg/kg, 70mg/kg, the 40mg/kg) intramuscular injection of the daily dosage of clinical plan of total alkaloids preparation as rat, every day 1 time, continuous 13 weeks, the variation of vital signs such as activity, appetite, defecation and the breathing of observation rat, heart rate, carry out detections such as hematology, serum biochemistry, dead rat carries out obduction and pathologic finding.The result shows that the high dose group rat has 2 death after 10 weeks, and pathological section raises, and causes minority rat injection site the scleroma of diameter less than 1.5cm to occur, recovers normal substantially after 2 weeks of drug withdrawal.All the other indexs there is no significant change.
The safety experiment of total alkaloids preparation adopts experimental techniques such as muscle irritation test, hypersensitive test, hemolytic test, pyrogen test, to estimate the safety of total alkaloids preparation.The result shows that the intramuscular injection of total alkaloids injection does not cause Cavia porcellus allergy to tame rabbit muscle nonirritant, and no hemolytic reaction does not contain pyrogen.Prompting total alkaloids preparation safety meets the requirement of injecting drug use.
Below by embodiment anticancer total alkaloids preparation of the present invention and preparation method thereof is described further.
Embodiment 1:
Get Herba Macleayae Cordatae medical material 1kg, be cut into the 1-2cm segment, add the immersion of 8 premium on currency, transfer solution PH to 2.0-3.0 with hydrochloric acid, often stir, soak 16-24h, filter pressing gets 5 liters of extracts; The aqueous solution soaking 16h that it is 2.0-3.0 that filtering residue adds 5 liters of PH again, filter pressing gets 5 liters of extracts.Merge extract twice, add concentrated NaOH solution, regulate extract pH to 6.0-7.0, standing over night makes the alkaloid precipitation; Filter, collect filtrate; Add NaOH solution in filtrate, regulate filtrate pH to 9.0-9.5, and add NaCl, make that NaCl concentration reaches 3.8% in the filtrate, stir NaCl is dissolved fully, standing over night makes the alkaloid precipitation; Filter collecting precipitation, get the total alkaloids precipitate of plant origin of the present invention, yield (in the constant weight product) is 1.5%, and after testing, chelerythrine in the gained precipitate (in total alkali content) is 50%, and Sanguinarine content is 35%, and other composition is 15%.
The total alkaloids precipitation is washed with water to neutrality, dispersion suspension is in 10 times of water gagings, transfer pH of suspension to 2.0-3.0, precipitation is dissolved fully, solution is passed through cation exchange column, earlier be washed till neutrality with distilled water, be eluted to the inanimate object alkali reaction with ammonia-alcoholic solution then till, collect the common 300ml of eluent; Add the 300ml chloroform in eluent, thermal agitation left standstill phase-splitting after 15 minutes, told ammonia-ethanol phase, added the 300ml chloroform extraction more once; Merge chloroform extraction liquid twice, under 50 ℃, reclaim under reduced pressure chloroform to extract volume be original volume 1/3 till.Add the 200ml acidic aqueous solution in spissated chloroform extraction liquid, regulate mixed solution pH2.0-3.0, concuss mixed solution 15 minutes leaves standstill phase-splitting, tells water; Repeat aqueous solution extraction 1 time, merge 2 times aqueous solution; Regulator solution pH is 9.0-9.5, and standing over night makes the alkaloid precipitation, filters to such an extent that precipitate; Precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, make resolution of precipitate, add hydrochloric acid while hot and transfer pH to 4.0-5.0, cooling, standing over night makes the alkaloid crystallization; Filter crystallization.Crystallization promptly obtains the salt of the total alkaloids of plant origin of the present invention in 40 ℃ of following vacuum dryings, and yield is 0.8%, and wherein the chelerythrine salt content is 80%-99.8%, and the Sanguinarine salt content is 0%-15%, and other component content is 0%-5%.
The total alkaloids and the pharmaceutical preparation in the pharmaceutical preparation thereof of plant origin of the present invention can be solid preparations, comprise tablet, capsule, soft capsule, slow releasing capsule, slow releasing tablet and pellet.These dosage forms all are to measure total alkaloid salt as required to add pharmaceutic adjuvant and filler, abundant mix homogeneously, and the existing pharmaceutical equipment of application and common process tabletting, pelletize, dress (filling) capsule are realized.
In the total alkaloids of plant origin of the present invention and the pharmaceutical preparation of pharmaceutical preparation thereof, wherein the preparation of injection and infusion solution is that total alkaloid salt is dissolved with 10 times of waters for injection by metering, the injection water solublity pharmaceutic adjuvant that adds metering, transfer pH to proper range, after the microporous filter membrane ultrafiltration, restock water for injection is to ormal weight, by the fill of preparation specification or make further dried and realize.
Get the total alkaloid salt 100g of plant origin of the present invention, with the dissolving of 1000ml water for injection, add an amount of antioxidant, cosolvent, aminoacid and phosphate, the adjusting pH value is 4.0-5.0, after the ultrafiltration of 0.2um microporous filter membrane, replenish water for injection to the ormal weight sterile filling.The injection fill in ampoule bottle, every 2ml, every total alkaloids (in chelerythrine) 100-200mg that contains plant origin of the present invention, sealing by fusing, through after the assay was approved the packing promptly.The infusion solution fill is in suitable infusion bottle, and specification is 100ml, 150ml, 200ml, contains total alkaloids (in chelerythrine) 100mg, 150mg, the 200mg of plant origin of the present invention respectively, rolls lid, and through after the assay was approved, packing promptly.
The total alkaloids injectable powder method of preparation plant origin of the present invention is, getting plumepoppy total alkaloid salt is dissolved in 5 times of amount waters for injection, add the injection water solublity pharmaceutic adjuvant of metering, as antioxidant, cosolvent, aminoacid or glucose or dextran, adjust pH is 4.0-5.0, through ultrafiltration, in 40 ℃ of following vacuum dryings, be sub-packed in the cillin bottle every bottle of 200mg after the pulverizing, roll lid, packing promptly gets injectable powder after the assay was approved.
The total alkaloids lyophilized injectable powder of preparation plant origin of the present invention; the total alkaloid salt of getting plant origin of the present invention is dissolved in 5 times of amount waters for injection; add the injection medicinal auxiliary agent of water solublity of metering; as antioxidant; cosolvent; aminoacid or glucose or dextran; adjust pH is 4.0-5.0; through ultrafiltration; be sub-packed in the ampoule; every 1ml; contain total alkaloids (in the chelerythrine alkali salt) 100-200mg of plant origin of the present invention, after lyophilization, sealing by fusing; through after the assay was approved, packing promptly gets the lyophilized injectable powder of plant macleaya cordata total alkaloid of the present invention and preparation thereof.
Embodiment 2:
Get Herba Macleayae Cordatae medical material 2kg, be cut into the 1-2cm segment, put in the extracting container, add 8 liters of methanol, add 8g citric acid and 8ml hydrochloric acid subsequently in the extracting container, soak 16-24h, sucking filtration gets 5 liters of extracts; Add 5 liters of methanol again in container, replenish citric acid 5g, hydrochloric acid 5ml soaks 8-12h, and sucking filtration gets 5 liters of extracts.Merge extract twice, transferring pH with high alkali liquid is 6.0-7.0, and standing over night makes the alkaloid precipitation; Filter, collect filtrate; It is 9.0-9.5 that continuation is transferred filtrate pH with concentrated base, and adds 30% liquor zinci chloridi, makes that zinc oxide concentration reaches 2%-4% in the filtrate, standing over night makes the alkaloid precipitation, filters collecting precipitation, get the total alkaloids precipitate of plant origin of the present invention, yield is 1.8%-2.2%.
Get the total alkaloids of plant origin of the present invention, be washed to neutrality after, join in the aqueous solution that 800ml pH is 2.0-3.0, it is fully dissolved, form the total alkaloids saline solution.Solution by cation exchange resin column, is washed to neutrality, be eluted to the inanimate object alkali reaction with ammonia-alcoholic solution then till, collect eluent 400ml altogether.Add the 400ml chloroform in eluent, transferring mixed solution pH is 9.0-9.2, thermal agitation 10-15 minute, leave standstill phase-splitting, and tell ammonia-ethanol phase, add chloroform 400ml again, accent pH is 9.0-9.2, extraction is once again; Merge chloroform extraction liquid twice, the reclaim under reduced pressure chloroform is to 400ml; Each 400ml water that adds is transferred mixed solution pH2.0-3.0 in spissated chloroform extraction liquid, extracts 2 times; Merge water, the water transfer pH value of solution is 9.0-9.5, makes the alkaloid precipitation, filters to such an extent that precipitate; Precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, precipitation is dissolved fully, add hydrochloric acid solution while hot, transfer pH to 4.0-5.0, standing over night makes the alkaloid crystallization, filter crystallization; Vacuum drying promptly gets the total alkaloid salt of plant origin of the present invention.
The total alkaloids preparation of pharmaceutical formulations method of plant origin of the present invention is with embodiment 1.
Embodiment 3:
Get Eomecon chionantha Hance medical material 1kg, be cut into the 1-2cm segment, add the immersion of 8 premium on currency, transfer pH value of solution to 2.0-3.0 with hydrochloric acid, often stir, soak 16-24h, filter pressing gets 5 liters of extracts; The aqueous solution soaking 16h that it is 2.0-3.0 that filtering residue adds 5 liters of pH again, filter pressing gets 5 liters of extracts.Merge extract twice, add dense Na
2CO
3Solution is regulated extract pH to 6.0-7.0, and standing over night makes the alkaloid precipitation; Filter, collect filtrate; In filtrate, add Na
2CO
3Solution is regulated filtrate pH to 9.0-9.5, and adds NaCl, makes that NaCl concentration reaches 3.8% in the filtrate, stirs NaCl is dissolved fully, and standing over night makes the alkaloid precipitation; Filter collecting precipitation, get the total alkaloids precipitate of plant origin of the present invention, yield (in the constant weight product) is 1.5%, and after testing, chelerythrine in the gained precipitate (in total alkali content) is 50%, and Sanguinarine content is 25%, and other composition is 25%.
The total alkaloids precipitation is washed with water to neutrality, dispersion suspension is in 10 times of water gagings, transfer pH of suspension to 2.0-3.0, precipitation is dissolved fully, solution is passed through cation exchange column, earlier be washed till neutrality with distilled water, be eluted to the inanimate object alkali reaction with ammonia-alcoholic solution then till, collect the common 300ml of eluent; Add the 300ml chloroform in eluent, accent pH is 9.0-9.5, and thermal agitation left standstill phase-splitting after 15 minutes, told ammonia-ethanol phase, added the 300ml chloroform extraction more once; Merge chloroform extraction liquid twice, under 50 ℃, reclaim under reduced pressure chloroform to extract volume be original volume 1/3 till.Add the 200ml acidic aqueous solution in spissated chloroform extraction liquid, regulate mixed solution pH2.0-3.0, concuss mixed solution 15 minutes leaves standstill phase-splitting, tells water; Repeat aqueous solution extraction 1 time, merge 2 times aqueous solution; Regulator solution pH is 9.0-9.5, and standing over night makes the alkaloid precipitation, filters to such an extent that precipitate; Precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, make resolution of precipitate, add hydrochloric acid while hot and transfer PH to 4.0-5.0, cooling, standing over night makes the alkaloid crystallization; Filter crystallization.Crystallization promptly obtains the salt of the total alkaloids of plant origin of the present invention in 40 ℃ of following vacuum dryings, and yield is 0.9%, and wherein the chelerythrine salt content is 80%-99.8%, and the Sanguinarine salt content is 0%-15%, and other component content is 0%-5%.
The preparation method of the total alkaloids pharmaceutical preparation of plant origin of the present invention of the present invention is with embodiment 1.
Embodiment 4:
Get Herba Chelidonii medical material 2kg, be cut into the 1-2cm segment, put in the extracting container, add 8 liters of methanol, add 8g citric acid and 8ml hydrochloric acid subsequently in the extracting container, soak 16-24h, sucking filtration gets 5 liters of extracts; Add 5 liters of methanol again in container, replenish citric acid 5g, hydrochloric acid 5ml soaks 8-12h, and sucking filtration gets 5 liters of extracts.Merge extract twice, transferring pH with high alkali liquid is 6.0-7.0, and standing over night makes the alkaloid precipitation; Filter, collect filtrate; It is 9.0-9.5 that continuation is transferred filtrate pH with concentrated base, and adds 30% ZnCl solution, makes that ZnCl concentration reaches 2%-4% in the filtrate, and standing over night makes the alkaloid precipitation, filters collecting precipitation, the total alkaloids precipitate, yield is 1.7%-2.1%.
Get total alkaloids, be washed to neutrality after, join in the aqueous solution that 800ml pH is 2.0-3.0, it is fully dissolved, form the total alkaloids saline solution.Solution by cation exchange resin column, is washed to neutrality, be eluted to the inanimate object alkali reaction with ammonia-alcoholic solution then till, collect eluent 400ml altogether.Add the 400ml chloroform in eluent, transferring mixed solution pH is 9.0-9.2, thermal agitation 10-15 minute, leave standstill phase-splitting, and tell ammonia-ethanol phase, add chloroform 400ml again, accent pH is 9.0-9.2, extraction is once again; Merge chloroform extraction liquid twice, the reclaim under reduced pressure chloroform is to 400ml; Each 400ml water that adds is transferred mixed solution pH2.0-3.0 in spissated chloroform extraction liquid, extracts 2 times; Merge water, the water transfer pH value of solution is 9.0-9.5, makes the alkaloid precipitation, filters to such an extent that precipitate; Precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, precipitation is dissolved fully, add hydrochloric acid solution while hot, transfer pH to 4.0-5.0, standing over night makes the alkaloid crystallization, filter crystallization; Vacuum drying promptly gets total alkaloid salt.
The total alkaloids preparation of pharmaceutical formulations method of plant origin of the present invention is with embodiment 1.
Embodiment 5:
Get Radix Toddaliae Asiaticae medical material 2kg, be cut into the 0.5-1cm segment, put in the extracting container, add 8 liters of methanol, add 8g citric acid and 8ml hydrochloric acid subsequently in the extracting container, soak 16-24h, sucking filtration gets 5 liters of extracts; Add 5 liters of methanol again in container, replenish citric acid 5g, hydrochloric acid 5ml soaks 8-12h, and sucking filtration gets 5 liters of extracts.Merge extract twice, transferring pH with high alkali liquid is 6.0-7.0, and standing over night makes the alkaloid precipitation; Filter, collect filtrate; It is 9.0-9.5 that continuation is transferred filtrate pH with concentrated base, and adds 30% NaCl solution, makes that NaCl concentration reaches 2%-4% in the filtrate, and standing over night makes the alkaloid precipitation, filters collecting precipitation, the total alkaloids precipitate, yield is 1.9%-2.3%.
Get total alkaloids, be washed to neutrality after, join in the aqueous solution that 800ml pH is 2.0-3.0, it is fully dissolved, form the total alkaloids saline solution.Solution by cation exchange resin column, is washed to neutrality, be eluted to the inanimate object alkali reaction with ammonia-alcoholic solution then till, collect eluent 400ml altogether.Add the 400ml chloroform in eluent, transferring mixed solution pH is 9.0-9.2, thermal agitation 10-15 minute, leave standstill phase-splitting, and tell ammonia-ethanol phase, add chloroform 400ml again, accent pH is 9.0-9.2, extraction is once again; Merge chloroform extraction liquid twice, the reclaim under reduced pressure chloroform is to 400ml; Each 400ml water that adds is transferred mixed solution pH2.0-3.0 in spissated chloroform extraction liquid, extracts 2 times; Merge water, the water transfer pH value of solution is 9.0-9.5, makes the alkaloid precipitation, filters to such an extent that precipitate; Precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, precipitation is dissolved fully, add hydrochloric acid solution while hot, transfer pH to 4.0-5.0, standing over night makes the alkaloid crystallization, filter crystallization; Vacuum drying promptly gets the total alkaloid salt of plant origin of the present invention.
The total alkaloids preparation of pharmaceutical formulations method of plant origin of the present invention is with embodiment 1.
Embodiment 6:
Get Herba Chelidonii and Eomecon chionantha Hance medical material 2kg, be cut into the 0.5-1cm segment, put in the extracting container, add 8 liters of methanol, add 8g citric acid and 8ml hydrochloric acid subsequently in the extracting container, soak 16-24h, sucking filtration gets 5 liters of extracts; Add 5 liters of methanol again in container, replenish citric acid 5g, hydrochloric acid 5ml soaks 8-12h, and sucking filtration gets 5 liters of extracts.Merge extract twice, transferring pH with high alkali liquid is 6.0-7.0, and standing over night makes the alkaloid precipitation; Filter, collect filtrate; It is 9.0-9.5 that continuation is transferred filtrate pH with concentrated base, and adds 30% NaCl solution, makes that NaCl concentration reaches 2%-4% in the filtrate, and standing over night makes the alkaloid precipitation, filters collecting precipitation, the total alkaloids precipitate, yield is 2.1%-2.5%.
Get total alkaloids, be washed to neutrality after, join in the aqueous solution that 800ml pH is 2.0-3.0, it is fully dissolved, form the total alkaloids saline solution.Solution by cation exchange resin column, is washed to neutrality, be eluted to the inanimate object alkali reaction with ammonia-alcoholic solution then till, collect eluent 400ml altogether.Add the 400ml chloroform in eluent, transferring mixed solution pH is 9.0-9.2, thermal agitation 10-15 minute, leave standstill phase-splitting, and tell ammonia-ethanol phase, add chloroform 400ml again, accent pH is 9.0-9.2, extraction is once again; Merge chloroform extraction liquid twice, the reclaim under reduced pressure chloroform is to 400ml; Each 400ml water that adds is transferred mixed solution pH2.0-3.0 in spissated chloroform extraction liquid, extracts 2 times; Merge water, the water transfer pH value of solution is 9.0-9.5, makes the alkaloid precipitation, filters to such an extent that precipitate; Precipitation is joined in 3 times of amount ethanol, be heated to 40-70 ℃, precipitation is dissolved fully, add hydrochloric acid solution while hot, transfer pH to 4.0-5.0, standing over night makes the alkaloid crystallization, filter crystallization; Vacuum drying promptly gets the total alkaloid salt of plant origin of the present invention.
The total alkaloids preparation of pharmaceutical formulations method of plant origin of the present invention is with embodiment 1.
Claims (2)
1. method of extracting the total alkaloids with active anticancer from Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance or Radix Toddaliae Asiaticae is characterized in that:
A. get Herba Macleayae Cordatae, Herba Chelidonii, Eomecon chionantha Hance or Radix Toddaliae Asiaticae medical material, be cut into the segment of 0.5-2cm, add the acid solution immersion 16-24h that 4-8 doubly measures pH2.0-3.0, filter pressing gets extract;
B. the acid solution that adds the pH 2.0-3.0 that 4-6 doubly measures in filtering residue again soaks 8-16h, and filter pressing gets extract;
C. merge extract twice, to 6.0-7.0, standing over night makes the alkaloid precipitation with the aqueous slkali adjust pH;
D. filter, collect filtrate;
E. filtrate continues to transfer pH to 9.0-9.5 with aqueous slkali, and adds saline solution, makes the salinity in the filtrate reach 3%-4%, and standing over night makes the alkaloid precipitation; Filter, collecting precipitation gets the total alkaloids precipitate.
2. method according to claim 1, wherein the total alkaloids precipitate adopts ion exchange resin absorption and solvent-extracted method further to purify, and the method for its purification is:
A. the total alkaloids precipitation is washed to neutrality, adds 10-20 and doubly measure the acid solution that pH is 2.0-3.0, make fully dissolving of precipitation;
B. solution is passed through cation exchange resin column, be washed to neutrality,, collect eluent with ammonia-alcoholic solution eluting;
C. add equal amounts of chloroform in eluent, adjusting pH is 9.0-9.5, and concuss 15min leaves standstill phase-splitting, tells ammonia-ethanol phase, adds the equal amounts of chloroform extraction more once, merges chloroform extraction liquid twice;
D. under 40-70 ℃, the reclaim under reduced pressure chloroform is to original volume 1/2-1/4;
E. in the chloroform concentrated extract, add the equivalent acidic aqueous solution, transfer pH value of solution 2.0-3.0, extract 2 times, merge water;
F. water transfer phase pH is 9.0-9.5, and standing over night makes the alkaloid precipitation, filters to such an extent that precipitate;
G. precipitation is joined 3 times and measure in the ethanol, be heated to 40-70 ℃, make resolution of precipitate, add hydrochloric acid solution while hot and transfer pH4.0-5.0, cooling, standing over night makes the alkaloid crystallization, filters, and collects crystallization, and dry under 40 ℃, promptly.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB03123772XA CN1292750C (en) | 2003-06-02 | 2003-06-02 | Total alkaloid composition from plant and its pharmaceutical preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB03123772XA CN1292750C (en) | 2003-06-02 | 2003-06-02 | Total alkaloid composition from plant and its pharmaceutical preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1457780A CN1457780A (en) | 2003-11-26 |
| CN1292750C true CN1292750C (en) | 2007-01-03 |
Family
ID=29430350
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB03123772XA Expired - Fee Related CN1292750C (en) | 2003-06-02 | 2003-06-02 | Total alkaloid composition from plant and its pharmaceutical preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1292750C (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100366621C (en) * | 2005-08-29 | 2008-02-06 | 长沙世唯科技有限公司 | Plumepoppy total alkaloid salt and its prepn and application |
| CN101732417B (en) * | 2010-02-09 | 2011-11-09 | 曾建国 | Preparation method and application of ion pair mixture of macleaya cordata total alkaloid |
| CN101904849A (en) * | 2010-08-19 | 2010-12-08 | 苏州大学 | Application of sanguinarine for preparing tumour radiotherapy sensitized medicine |
| CN104069218A (en) * | 2013-03-29 | 2014-10-01 | 上海中医药大学 | Preparation method and usage of Asiatic toddalia root total alkaloid effective part extract |
| CN105267309A (en) * | 2015-11-24 | 2016-01-27 | 湖南汉清生物技术有限公司 | Oral liquid medicine for canine parvovirus and preparation method thereof |
| CN110141605A (en) * | 2019-04-19 | 2019-08-20 | 南京康齐生物科技有限公司 | A kind of preparation method of snowpoppy total alkaloid nanoparticle |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4767861A (en) * | 1986-01-28 | 1988-08-30 | Vipont Laboratories | Recovery of benzo-c-phenanthridine alkaloids |
| US4769452A (en) * | 1986-02-07 | 1988-09-06 | Vipont Laboratories, Inc. | Production of purity benzo-c-phenanthridine alkaloid salts |
| US4818533A (en) * | 1985-07-09 | 1989-04-04 | Vipont Pharmaceutical, Inc. | Production of high purity alkaloids |
| CN1255061A (en) * | 1997-03-25 | 2000-05-31 | 阿奇发展有限公司 | Use of chelerythrine and radiation for tumor therapy |
-
2003
- 2003-06-02 CN CNB03123772XA patent/CN1292750C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4818533A (en) * | 1985-07-09 | 1989-04-04 | Vipont Pharmaceutical, Inc. | Production of high purity alkaloids |
| US4767861A (en) * | 1986-01-28 | 1988-08-30 | Vipont Laboratories | Recovery of benzo-c-phenanthridine alkaloids |
| US4769452A (en) * | 1986-02-07 | 1988-09-06 | Vipont Laboratories, Inc. | Production of purity benzo-c-phenanthridine alkaloid salts |
| CN1255061A (en) * | 1997-03-25 | 2000-05-31 | 阿奇发展有限公司 | Use of chelerythrine and radiation for tumor therapy |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1457780A (en) | 2003-11-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1857343A (en) | Chinese medicine composition for preventing and treating viral cold and throat disease and its preparing method | |
| CN1257969C (en) | Polygonum capitatum extract and preparation method and application thereof | |
| CN1127648A (en) | Chinese medicine composition with antipyretic and antidote functions and use thereof | |
| CN1049973A (en) | Has new medicinal preparation of antitumaous effect and preparation method thereof | |
| CN1292750C (en) | Total alkaloid composition from plant and its pharmaceutical preparation | |
| CN1299702C (en) | Preparation of medicine preparation of orthopedics department and its use | |
| CN1748748A (en) | Little leaf deervetch herb preparation for pregnant woman and new preparing method | |
| CN101411779A (en) | Chinese medicine effective component composition for treating liver cancer and method for preparing the same | |
| CN1823982A (en) | Chinese medicinal preparation for liver soothing and speen fortifying and its manufacturing method | |
| CN1903351A (en) | Funing gel prepn. and its prepn. method | |
| CN1686424A (en) | Medicinal composition containing scutellaria and bupleurum and its preparation method | |
| CN1233401C (en) | Agastache capsule for restoring healthy energy andits prepn and application | |
| CN1698820A (en) | Dripping pills with jaundice eliminating liver protecting functions and its preparation method | |
| CN1272026C (en) | Medicinal composition for treating cardiocerebral vasculr disease and its preparing method | |
| CN1742926A (en) | Zuoci preparation for deaf and new preparing method thereof | |
| CN1278731C (en) | Compound bupleurum root medicinal preparation | |
| CN1824099A (en) | Diffusing-freeing lung rectifying medicinal preparation and its new preparation method | |
| CN1406585A (en) | Medicinal composition for viral myocarditis | |
| CN1785212A (en) | Novel cinobufogenin freeze-drying powder injection, its prepn. method and quality control method | |
| CN1762362A (en) | Medicinal composition with red sage root component for treating cardiovascular and cerebrovascular disease and its application | |
| CN1650971A (en) | Preparation method of rape myricyl powder effective portion group | |
| CN1634509A (en) | Yitongshu drop pill for treating pain and its preparation method | |
| CN1079396A (en) | Intelligence-enhancing pharmaceutical preparation | |
| CN1278698C (en) | Freeze dried injection of seeding of autumnal sowthistle-leaf ixeris and preparation method thereof | |
| CN1207002C (en) | Antineoplastic composite and its prepn process and use |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |