CN1772026A - Whorlleaf stonecrop herb extract and its extraction process and prepn - Google Patents

Whorlleaf stonecrop herb extract and its extraction process and prepn Download PDF

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CN1772026A
CN1772026A CN 200510115124 CN200510115124A CN1772026A CN 1772026 A CN1772026 A CN 1772026A CN 200510115124 CN200510115124 CN 200510115124 CN 200510115124 A CN200510115124 A CN 200510115124A CN 1772026 A CN1772026 A CN 1772026A
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extract
herba senecionis
preparation
senecionis cannabifolii
water
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CN100367977C (en
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吴立军
高慧媛
刘传贵
金立群
朱继忠
关家华
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HUAKANG PHARMACEUTICAL CO Ltd JILIN
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HUAKANG PHARMACEUTICAL CO Ltd JILIN
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Abstract

The present invention is one kind one whorlleaf stonecrop herb extract and its extraction process and preparation. The whorlleaf stonecrop herb extract contains few alkaloid components, and is prepared through the following steps: water extraction, alcohol precipitation, and processing in cationic exchange resin column. The present invention also provides medicine preparation with the extract as the active component, and the medicine preparation is oral preparation, especially micro pill preparation.

Description

A kind of Herba Senecionis cannabifolii extract, its extracting method and preparation
Technical field:
The present invention relates to a kind of Chinese medicine extraction method, particularly a kind of extracting method of Chinese medicine Herba Senecionis cannabifolii, and the extract and the pharmaceutical preparation thereof that obtain with this method.
Background technology:
Upper respiratory tract infection (flu) is the frequently-occurring disease commonly encountered diseases.Be the modal a kind of infectious disease of respiratory tract, the patient is of all ages, sex, occupation and regional, and all can fall ill the whole year, the winter-spring season pilosity, people can have morbidity for several times in 1 year, and can cause severe complication, sickness rate height not only, and be multiple Disease Inducement.Upper respiratory tract infection 90% is viral infection simultaneously, and performance Chinese medicine antivirus action is imperative.
It is 3.82% that the chronic tracheitis China whole nation seventies 6000 ten thousand people generally investigate average attack rate, and prevalence increases with age, and patient's prevalence is up to 15% or more more than 50 years old.Boreal climate cold is higher than south, and the industrial and mineral area is higher than general city, has a strong impact on people's physical and mental health.Though this is had research, still there is not specific short so far, though there are serious toxic and side effects of certain curative effect and drug resistance to be difficult to overcome for the antibacterial Western medicine of antiinflammatory,
Herba Senecionis cannabifolii is the distinctive a kind of Chinese herbal medicine in Changbai Mountain, and the locals is ill, and oneself going up a hill gathers medicinal herbs endures water and drink custom, it is said that this medicine comes back to life, and returns the special effect of people's soul, the Herba Senecionis cannabifolii of gaining the name then.Herba Senecionis cannabifolii has cough-relieving, reduces phlegm, relieving asthma, and antiinflammatory, domestic somebody is with these medicine treatment emphysema, and Japan treats the report of pulmonary carcinoma in addition.Existing Herba Senecionis cannabifolii electuary has heat clearing away to eliminate the phlegm, the effect of cough-relieving.Be used for the chronic bronchitis cough.Its preparation method is deferred to the Chinese medicine standard that country formulates, and the preparation method in the standard is as follows:
[method for making] gets Herba Senecionis cannabifolii 1000g, decocts with water three times, and 2 hours for the first time, each 1 hour collecting decoction of second and third time filtered, and filtrate is condensed into cream, and adding sucrose is an amount of, and mixing is made granule, and drying is made 1000g, promptly.
Though Herba Senecionis cannabifolii electuary, tablet, oral liquid listing are arranged at present, be the simple extraction of medical material, effective site is indeterminate, and characteristic toxic substance alkaloid is not studied, and quality is wayward.The present invention makes new dosage form by with the Herba Senecionis cannabifolii deep processing, pellet preparations form particularly, the collection antiviral, antibacterial be one, the effect of have cough-relieving again, reduce phlegm, relievining asthma.The present invention extracts active ingredient earlier, removes total alkaloids by cation exchange resin subsequently, and it is many to obtain containing effective ingredient, and the new extract that by-product is few is made micropill through the galenic pharmacy technology then, and toxicity obviously reduces.
Summary of the invention:
The invention provides the new extract of a kind of Herba Senecionis cannabifolii, this extract is characterised in that, does not contain or contain hardly alkaloid component.
The present invention also provides the preparation method of Herba Senecionis cannabifolii extract of the present invention, and these method process following steps: water is carried, precipitate with ethanol, last cation exchange resin column.
The present invention also provides with the pharmaceutical preparation of extract of the present invention as the active constituents of medicine preparation, particularly oral pharmaceutical preparation, preferably pellet preparations.
The preparation method of Herba Senecionis cannabifolii extract of the present invention, wherein said water is carried, and is that process decocts with water 1-4 time, and the amount that at every turn adds entry is 5-15 times, each 1-3 hour.
Described precipitate with ethanol, be with water extract concentrate obtain clear paste after, add the ethanol of 50-100%, left standstill 6-24 hour, treat that precipitation separates out after-filtration and fall precipitation, reclaim ethanol, concentrate and obtain clear paste.
Described upward cation exchange resin column is that the clear paste that will obtain behind the precipitate with ethanol joins cation exchange resin column, adsorbs 10-60 minute, and the water eluting obtains water elution liquid then, and this eluent drying obtains extract of the present invention.。Wherein said resin is any cation exchange resin, and preferably polystyrene cation exchange resin, or acrylic acid type cation exchange resin, its consumption are clear paste: resin=1: 2-2: 1.
The form of extract that the present invention obtains can be the material of extractum form, can be that dry extract also can be a fluid extract, makes different concentration according to the different needs decision of preparation.
The most preferred preparation method of the present invention is as follows:
Get Herba Senecionis cannabifolii, decoct with water three times, add water 12-9 respectively doubly, 10-8 doubly, 10-8 doubly, each 1-1.5 hour, collecting decoction filtered, filtrate is concentrated into the clear paste that relative density is 1.10~1.30g/ml (80 ℃ of survey), puts coldly, adds 95% ethanol, mixing, to pure relative density 75%, left standstill precipitation 12 hours, filter, decompression filtrate recycling ethanol, being concentrated into relative density is the clear paste of 1.10~1.20g/ml (60 ℃ of surveys), puts cold, amount with clear paste: resin=1-2: 1-2, last cation exchange resin column adsorbs 20-40min on resin column, then the water eluting, collect water elution part spray drying, promptly get extract.
The extract that above method obtains, through the alkaloid detecting instrument detection of prior art established practice, wherein alkaloid is less than 0.5%.
Institute of the present invention art pharmaceutical preparation, be the extract that does not contain or contain alkaloid component hardly that said method is obtained as active constituents of medicine, through the materia medica routine techniques, be prepared into the pharmaceutical preparation of any form,
In preparation process, the weight of Herba Senecionis cannabifolii is calculated with crude drug, and this weight composition can be made into 1000 doses of pharmaceutical preparatioies.Described 1000 doses of fingers, the final drug preparation of making, as make 1000 of capsule preparations, 1000 in tablet, granule 1000 grams, oral liquid 1000ml etc., also can make big packing as granule, as the 100-500 bag, specifically can be 100 bags, 125 bags, 200 bags, 250 bags, 500 bags etc., every bag can be used as taking dose 1-2 time.More than form, can be made into the preparation of 50-1000 taking dose,, make 1000, make 1000 of capsules as tablet, each taking dose can be the 1-20 sheet/or, can take 50-1000 time altogether.As granule, make 125 bags, take the 1-2 bag at every turn, can take 62.5-125 time altogether.
More than form being by weight as proportioning, can increasing or reduce according to corresponding proportion when producing, can be unit with the kilogram as large-scale production, or is unit with the ton.
Pharmaceutically active substance in the Chinese medicine preparation of the present invention, its shared percentage by weight in preparation can be 0.1-99.9%, all the other are the medicine acceptable carrier.Pharmaceutical preparation of the present invention exists with unit dosage form, and described unit dosage form is meant the unit of preparation, as every of tablet, capsular every capsules, every of injection etc., in the unit dose, the amount that contains active substance is 5-800mg, preferably 20-500mg.
Chinese medicine preparation of the present invention can be any pharmaceutically useful dosage form, these dosage forms comprise: tablet, capsule, oral liquid, syrup, granule, pill, pellet, powder, unguentum, sublimed preparation, injection, suppository, cream, spray, drop pill, patch, slow releasing preparation, controlled release preparation, preferably pellet.
Pharmaceutical preparation of the present invention, the preparation of its oral administration can contain excipient commonly used, such as binding agent, filler, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent, can carry out coating to tablet in case of necessity.
The filler that is suitable for comprises cellulose, mannitol, lactose and other similar filler.Suitable disintegrating agent comprises starch, polyvinylpyrrolidone and starch derivatives, for example sodium starch glycollate.Suitable lubricant, for example magnesium stearate.The acceptable wetting agent of appropriate drug comprises sodium lauryl sulphate.
Can fill by mixing, the method that tabletting etc. are commonly used prepares solid oral composition.Mix repeatedly active substance is distributed in those compositionss of a large amount of filleies of whole use.
The form of oral liquid for example can be aqueous or oily suspensions, solution, Emulsion, syrup or elixir, perhaps can be a kind of available water before use or other suitable composite dry products of carrier.This liquid preparation can contain conventional additive, such as suspending agent, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl-cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenation edible fat, emulsifying agent, for example lecithin, anhydro sorbitol monooleate or arabic gum; Non-aqueous carrier (they can comprise edible oil), for example almond oil, fractionated coconut oil, such as oily ester, propylene glycol or the ethanol of the ester of glycerol; Antiseptic, for example para hydroxybenzene methyl ester or propyl p-hydroxybenzoate or sorbic acid, and if desired, can contain conventional flavouring agent or coloring agent.
For injection, the liquid unit dosage forms of preparation contains active substance of the present invention and sterile carrier.According to carrier and concentration, this chemical compound can be suspended or dissolving.The preparation of solution is normally by being dissolved in active substance in a kind of carrier filter-sterilized before it is packed into a kind of suitable bottle or ampoule, sealing then.For example a kind of local anesthetic of adjuvant, antiseptic and buffer agent also can be dissolved in this carrier.In order to improve its stability, can be after the bottle of packing into that this compositions is freezing, and under vacuum, water is removed.
Pharmaceutical preparation of the present invention, when being prepared into medicament, optionally add suitable medicine acceptable carrier, described medicine acceptable carrier is selected from: mannitol, sorbitol, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin C, the EDTA disodium, EDTA calcium sodium, the alkali-metal carbonate of monovalence, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulphuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, the phospholipid material, Kaolin, Pulvis Talci, micropowder silica gel, magnesium carbonate, calcium stearate, magnesium stearate etc.
Preparation of the present invention is determined usage and dosage according to patient's situation in use, but obeys 1-4 time every day, each 1-20 agent, as: 1-20 grain or sheet.
Below by pharmacodynamics test beneficial effect of the present invention is described:
One, Herba Senecionis cannabifolii extract antiviral drug effect research report
Result of the test
1. the TCID of various viruses 50
Influenza virus FM 1: Log TCID 50 = - 2 + 0 . 5 - 100 + 100 + 50 100 = - 4
CVB 3 Log TCID 50 = - 2 + 0 . 5 - 100 + 100 + 100 + 50 100 = - 5
HSV-1SM 44 Log TCID 50 = - 2 + 0.5 - 100 + 100 + 100 + 30 100 = - 4.8
AdV-3: Log TCID 50 = - 2 + 0.5 - 100 + 100 + 50 100 = - 4
RSV: Log TCID 50 = - 2 + 0.5 - 100 + 100 + 100 + 50 100 = - 5
Parainfluenza virus: Log TCID 50 = - 2 + 0.5 - 100 + 100 + 50 100 = - 4
Influenza B virus: Log TCID 50 = - 2 + 0.5 - 100 + 100 + 50 100 = - 4
2. medicine pair cell toxicity test result
1) medicine pair cell toxicity test result
Herba Senecionis cannabifolii extract is to Hep-2, Hela cell maximal non-toxic concentration (TC 0) be 12.50g/L, half toxic concentration (TC 50) be 98.53g/L, concrete amount effect relation curve sees Fig. 1 for details.SHUANGHUANLIAN is to Hep-2, Hela cell
Maximal non-toxic concentration (TC 0) be 18.75g/L, half toxic concentration (TC 50) be 90.61g/L.
2) medicine is to the animal toxicity measurement result
Herba Senecionis cannabifolii extract mouse stomach administration half lethal dose (LD 50) be 2662g crude drug/kg, the 95% credible 2134~3320g/kg that is limited to.
3. medicine is to pathological changes caused by virus protective effect result
Infecting 30-100TCID 50The situation of 6 kinds of viruses under, Herba Senecionis cannabifolii extract has than its cytopathic effect of obvious suppression, to Coxsackie B virus influenza virus, parainfluenza virus, respiratory syncytial virus, herpes simplex virus I-type in 6.25g/L~12.5g/L concentration 3Not obvious with the adenovirus type III effect.Its exercising result sees table 1,2 for details.
Table 1 Herba Senecionis cannabifolii extract is to the protective effect of pathological changes caused by virus effect
Strain Infective dose SHUANGHUANLIAN Herba Senecionis cannabifolii extract Virus control The cell contrast
TCID 5 18.75g/L 12.5g/L 6.25g/L 3.125g/L
Influenza FM 1 CVB 3AdV-3 parainfluenza RSV HSV-I 100 30 100 100 50 30 - + - - - + + - - + ++ ++ + +++ +++ + ++++ ++++ ++++ ++++ ++++ ++++ - - - - - -
Annotate :-acellular pathological changes shown; ± be shown with and delay cytopathic effect; + show the cytopathy below 1/4; ++ show that 1/4~1/2 cell has pathological changes; The cell of +++show 1/2~3/4 has pathological changes; ++ ++ show that the cell more than 3/4 has pathological changes.
Table 2 Herba Senecionis cannabifolii extract suppresses medium effective concentration and the therapeutic index that cell virus infects
Influenza FM 1 CVB3 AdV-3 Parainfluenza RSV HSV-I
Herba Senecionis cannabifolii extract IC 50(g/L) TI 3.388 29.082 33.467 2.944 19.317 5.101 1.765 55.824 1.965 50.142 2.714 36.304
4. medicine is to the influence of mice influenza virus property pneumonia
1) Herba Senecionis cannabifolii extract adopts the EXCEL statistical software to the result that influences of mice pneumonia, carries out two groups of mean t checks, sees table 3,4 for details.Herba Senecionis cannabifolii extract all has significant therapeutic effect to first type and Influenza B virus pneumonia when 9.430g/kg/d and 28.300g/kg/d dosage.
Table 3 Herba Senecionis cannabifolii extract is to the effect of mice influenza A virus pneumonia (X ± SD)
Group Dosage (g/kg/d) Animal (only) Lung exponential quantity (g/g * 100%) Pneumonopathy becomes and alleviates (%) The P value
Virus control normal control SHUANGHUANLIAN Herba Senecionis cannabifolii extract - - 0.78 28.300 9.430 10 10 10 10 10 1.604±0.177 1.314±0.136 1.408±0.096 1.387±0.132 1.429±0.096 - - 12.22 13.53 10.91 - - <0.01 <0.01 <0.05
2.830 0.943 10 10 1.488±0.119 1.503±0.111 7.23 6.30 >0.05 >0.05
Table 4 Herba Senecionis cannabifolii extract is to the effect of mice Influenza B virus pneumonia (X ± SD)
Group Dosage (g/kg/d) Animal (only) Lung exponential quantity (g/g * 100%) Pneumonopathy becomes and alleviates (%) The P value
Virus control normal control SHUANGHUANLIAN Herba Senecionis cannabifolii extract - - 0.78 28.300 9.430 2.830 0.943 10 10 10 10 10 10 10 1.700±0.186 1.314±0.136 1.458±0.102 1.443±0.068 1.499±0.156 1.573±0.116 1.674±0.132 - - 14.24 15.12 11.82 7.47 1.53 - - <0.01 <0.01 <0.05 >0.05 >0.05
2) pneumonopathy becomes histology and pathology testing result
Viral pneumonia model arrangement of mirrors is following as seen: interstitial lung hyperemia, edema and lymphocyte, monocyte infiltrations such as bronchus, bronchioles wall, alveolar wall, and alveolar wall broadening, alveolar is inflammatory reaction.Herba Senecionis cannabifolii extract treatment 28.300g/kg/d and the apparition of 9.430g/kg/d dosage group Mus pneumonopathy alleviate, and the lung tissue segment morphosis is normal, and optimal therapeutic dosage is 28.300g/kg.
3) virus antigen titer determination----hemagglutination test result in the mouse lung tissue
The hemagglutination test result asks for an interview table 5-6.
Table 5 Herba Senecionis cannabifolii extract is organized the influence of blood clotting titre to influenza A virus pneumonia mouse lung
Group Dosage (g/kg/d) Animal (only) The blood clotting titre (X ± S) Suppression ratio (%) P
Virus control normal control SHUANGHUANLIAN Herba Senecionis cannabifolii extract - - 0.78 28.300 9.430 2.830 0.943 10 10 10 10 10 10 10 32.40±3.50 0.00 24.00±4.00 22.00±3.65 28.60±3.53 30.20±3.33 31.00±3.30 - - 25.93 32.10 11.73 6.79 4.32 - - <0.01 <0.01 <0.05 >0.05 >0.05
Table 6 Herba Senecionis cannabifolii extract is organized the influence of blood clotting titre to Influenza B virus pneumonia mouse lung
Group Dosage (g/kg/d) Animal (only) The blood clotting titre (X ± S) Suppression ratio (%) P
Virus control normal control SHUANGHUANLIAN Herba Senecionis cannabifolii extract - - 0.78 28.300 9.430 2.830 0.943 10 10 10 10 10 10 10 33.20±3.43 0.00 23.40±3.43 22.60±3.53 29.20±3.55 30.40±3.37 31.00±3.16 - - 29.52 31.93 12.05 8.43 6.63 - - <0.01 <0.01 <0.05 >0.05 >0.05
From above-mentioned two tables as seen, Herba Senecionis cannabifolii extract treatment 28.300g/kg and 9.430g/kg dosage group, first type and Influenza B virus titre all are starkly lower than the virus control group in the Mus lung.
Conclusion: Herba Senecionis cannabifolii extract is in the external effect that obvious suppression influenza virus, parainfluenza virus, respiratory syncytial virus and herpes simplex virus I-type are arranged, and is not obvious to the inhibitory action of coxsackie B 3 viruses and adenovirus type III.The interior resisting virus result of the test shows that Herba Senecionis cannabifolii extract can effectively suppress the viral pneumonia due to first type and the B-mode influenza virus, compares significant difference with model control group.Remain further to be furtherd investigate about the antiviral mechanism of action of this medicine.
Two, the antibiotic results of pharmacodynamic test of Herba Senecionis cannabifolii extract
(1) in-vitro antibacterial test result of the test
Herba Senecionis cannabifolii extract sees Table 17 to the minimal inhibitory concentration (MIC) of antibacterial.
Table 17 Herba Senecionis cannabifolii extract is to the outer MIC measurement result of 6 kinds of bacterial bodies
Antibacterial The strain number Medicine MIC scope (g/L) MIC 50 (g/L) MIC 90 (g/L)
Staphylococcus aureus beta hemolytic streptococcus pneumococcus EHEC Klebsiella Pneumoniae pseudomonas aeruginosa 8 11 6 9 8 9 The two coptiss of the two coptis Herba Senecionis cannabifolii extracts of the two coptis Herba Senecionis cannabifolii extracts of the two coptis Herba Senecionis cannabifolii extracts of the two coptis Herba Senecionis cannabifolii extracts of the two coptis Herba Senecionis cannabifolii extracts of Herba Senecionis cannabifolii extract 62.5~125 75~300 62.5~125 37.5~300 31.25~62.5 18.75~300 62.5~125 37.5~300 62.5~125 37.5~150 62.5~125 150~300 62.5 75 62.5 37.5 31.25 62.5 62.5 37.5 62.5 37.5 62.5 150 125 300 125 300 6.25 300 125 300 125 150 125 300
Conclusion (of pressure testing)
Extracorporeal bacteria inhibitor test shows that Herba Senecionis cannabifolii extract has certain inhibitory action to streptococcus pneumoniae, to the inhibitory action of staphylococcus aureus, Pseudomonas aeruginosa, escherichia coli, beta hemolytic streptococcus, Klebsiella Pneumoniae relatively a little less than.
(2) antibacterial tests result in the body
Herba Senecionis cannabifolii extract sees Table 18 to the endogenous protective result of the test of streptococcus pneumoniae infection mice.
Table 18 Herba Senecionis cannabifolii extract is to the endogenous protective test of pneumococcal infection mice
Medicine Dosage (g/kg) Log10 dose Number of animals (only) Death toll (only) Mortality rate (%)
Herba Senecionis cannabifolii extract 28.300 9.430 2.830 1.4518 0.9745 0.4518 10 10 10 2 5 7 20 50 70
The SHUANGHUANLIAN normal saline 0.943 0.283 0.780 -0.0255 -0.5482 10 10 10 10 8 10 2 10 80 100 20 100
According to result of the test, adopt the Bliss software processes, ED 50For: 7.211g/kg
95% fiducial limit: Ig (1.9756 ± 1.2454)
95% credibility interval: (2.0754-25.0545)
95% fiducial limit: Ig (1.9756 ± 2.2858)
95% credibility interval: (0.7333-70.9117)
Conclusion
The in vivo test result shows that infection has significant protective effect, its ED to Herba Senecionis cannabifolii extract to the S. pneumoniae strains induced mice 50Be respectively 7.211g/kg, 2.0754-25.0545. with 95% fiducial limit
Three, other Pharmacodynamic test of active extract researchs
Result of the test
One, antiinflammatory test
1, the Herba Senecionis cannabifolii extract xylol causes the influence of Mice Auricle inflammation edema
Result's (seeing Table 20-1) shows, the ear swelling that xylol causes mice behind Herba Senecionis cannabifolii extract 8g/kg (low dose group), 16g/kg (middle dosage group), 32g/kg (high dose group) the administration 1h all has significant inhibitory effect (p<0.05, p<0.001, p<0.001), suppression ratio is respectively 16.1%, 24.9%, 38.0%, and action intensity and dosage have certain dependency.Positive drug Herba Senecionis cannabifolii granule 16g/kg also has obvious suppression effect (p<0.01) to ear swelling in this model, and suppression ratio is 23.7%.
Table 20-1 Herba Senecionis cannabifolii extract xylol cause mice ear influence (X ± SD, n=12)
Group Dosage (g/kg) Swelling degree (mg) Suppression ratio (%)
Model control group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 8 16 32 16 16.02±2.63 13.44±2.02 * 12.03±1.60 *** 9.93±2.44 *** 12.23±2.35 ** - 16.1 24.9 38.0 23.7
*P<0.05 *P<0.01 * *Compare with model control group p<0.001
2, the Herba Senecionis cannabifolii extract on Carrageenan causes the influence of rat toes swelling
Result's (seeing Table 20-2) shows that this model 4h after causing inflammation reaches the edema peak.Herba Senecionis cannabifolii extract is a produce effects causing scorching back 2h, compares with the solvent matched group, and Herba Senecionis cannabifolii extract 5.4,10.8,21.6g/kg all can significantly suppress the rat toes swelling (p<0.05, p<0.01, p<0.001) due to 1% carrageenin.Positive drug Herba Senecionis cannabifolii granule 10.8g/kg is causing scorching back 2h produce effects, and its effect can continue until and causes scorching back 6h (p<0.05, p<0.01).
Table 20-2 Herba Senecionis cannabifolii extract on Carrageenan cause the swelling of rat toes influence (X ± SD, n=12)
Group Dosage (g/kg) Different time points swelling degree (%) and inhibitory rate of intumesce
1h 2h 3h 4h 5h 6h
Control group Herba Senecionis cannabifolii extract Fanhuncao Granule 5.4 10.8 21.6 10.8 28.66±15.33 23.43±11.38 18.2% 23.12±4.81 19.3% 23.01±10.79 19.7% 23.62±5.10 17.6% 57.97±23.17 36.04±5.61 * 37.8% 34.27±9.68 * 40.9% 28.57±12.03 ** 50.7% 33.89±13.38 * 41.5% 72.87±25.12 50.43±11.58 * 30.8% 47.46±6.28 ** 34.9% 45.14±13.37 ** 38.1% 49.12±16..53 * 32.6% 91.95±29.23 61.16±11.25 * 33.5% 53.37±17.05 ** 42.0% 52.03±12.98 *** 43.4% 59.48±4.97 * 35.3% 79.58±30.43 53.97±15.14 * 32.2% 48.37±11.51 * 39.2% 45.92±14.05 ** 42.3% 53.05±11.12 * 33.3% 80.05±25.39 54.19±10.20 * 32.3% 48.34±10.83 * 39.6% 46.51±13.95 ** 41.9% 54.13±12.05 * 32.4%
*P<0.05 *Compare with matched group p<0.01
3, the inhibitory action of Herba Senecionis cannabifolii extract Chinese People's Anti-Japanese Military and Political College Mus granuloma induced by implantation of cotton pellets
Experimental result (seeing Table 20-3) shows that each dosage group Herba Senecionis cannabifolii extract has obvious suppression effect: p<0.05, p<0.01, p<0.001 to rat granuloma is swollen.
Table 20-3 Herba Senecionis cannabifolii extract to the bullate inhibitory action of rat granuloma (X ± SD, n=12)
Group Dosage (g/kg) Granuloma dry weight (mg) Granuloma suppression ratio (%)
Blank group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 5.4 10.8 21.6 10.8 95.2±25.8 68.2±21.0 * 25.9±11.8 ** 19.6±10.4 *** 32.3±13.2 ** - 28.3 72.8 79.4 66.1
Two, separate heat test
4, Herba Senecionis cannabifolii extract is to the influence of fever in rabbits due to the endotoxin
The result shows (seeing Table 20-4), and auricular vein injection endotoxin is after 1 hour, and rabbit obviously generates heat.Gastric infusion 0.5h, Herba Senecionis cannabifolii extract 2.81,5.62,11.24g/kg can obviously bring into play refrigeration function: p<0.05, p<0.01, p<0.001, and effect can continue 4 hours.Herba Senecionis cannabifolii granule 5.62g/kg also can obviously suppress the fervescence (p<0.05, p<0.01) of each time point rabbit.
5, Herba Senecionis cannabifolii extract is to the influence of rat fever due to the beer yeast suspension
The result shows (seeing Table 20-5), the subcutaneous injection beer yeast after 4 hours rat temperature obviously raise.Compare with model control group, Herba Senecionis cannabifolii extract 5.4g/kg (low dose group), 10.8g/kg (middle dosage group), 21.6g/kg (high dose group) gastric infusion 1.0h can significantly suppress pyrogenic action (p<0.05 of beer yeast, p<0.01, p<0.001), and refrigeration function can continue the long period; Herba Senecionis cannabifolii granule 10.8g/kg also can obviously suppress the fervescence (p<0.05, p<0.01) of rat at each time point.
Table 20-4 Herba Senecionis cannabifolii extract to endotoxin cause fever in rabbits influence (X ± SD, n=8)
Group Dosage (g/kg) Basal body temperature (℃) The pyrogenicity temperature (℃) Each time point temperature after the administration (℃)
0.5h 1h 2h 3h 4h 5h 6h
Model control group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii groups of grains - 2.81 5.62 11.24 5.62 38.6±0.3 38.6±0.2 38.6±0.3 38.6±0.4 38.6±0.2 40.1±0.4 39.9±0.5 40.0±0.3 40.1±0.3 40.2±0.3 40.3±0.2 39.9±0.4 * 39.8±0.1 ** 39.7±0.2 ** 39.8±0.2 ** 40.4±0.3 40.0±0.2 * 39.8±0.1 ** 39.4±0.2 *** 39.7±0.4 ** 40.7±0.3 40.2±0.3 * 39.6±0.1 *** 39.3±0.1 *** 40.1±0.3 * 40.2±0.2 39.9±0.2 * 39.6±0.3 ** 39.2±0.2 *** 39.8±0.2 * 39.5±0.3 39.3±0.3 39.2±0.2 * 38.9±0.1 ** 39.2±0.2 * 39.0±0.2 38.8±0.3 38.8±0.2 38.8±0.1 39.0±0.2 38.6±0.2 38.6±0.2 38.6±0.3 38.7±0.2 38.8±0.3
*P<0.05 *P<0.01 * *Compare with model control group p<0.001
Table 20-5 Herba Senecionis cannabifolii extract to beer yeast cause rat fever influence (X ± SD, n=12)
Group Dosage (g/kg) Basal body temperature (℃) The pyrogenicity temperature (℃) Each time point temperature after the administration (℃)
0.5h 1h 2h 3h 4h 5h 6h
Model control group Herba Senecionis cannabifolii extract group Herba Senecionis cannabifolii groups of grains - 5.4 10.8 21.6 10.8 37.5±0.2 37.4±0.2 37.3±0.2 37.3±0.3 37.4±0.4 39.0±0.2 39.0±0.2 39.0±0.2 39.0±0.3 38.9±0.3 39.4±0.2 39.0±0.4 * 38.9±0.2 ** 38.8±0.2 ** 39.0±0.2 ** 39.6±0.1 39.0±0.2 ** 38.9±0.1 ** 38.7±0.2 *** 38.8±0.3 ** 39.4±0.2 38.8±0.3 * 38.5±0.3 ** 38.3±0.2 *** 38.5±0.4 ** 39.2±0.2 38.7±0.4 * 38.4±0.2 ** 38.3±0.1 *** 38.6±0.3 * 39.0±0.2 38.6±0.4 * 38.4±0.2 ** 38.3±0.1 *** 38.7±0.2 * 38.9±0.4 38.5±0.4 * 38.4±0.3 * 38.3±0.2 ** 38.5±0.3 * 38.7±0.2 38.5±0.3 38.4±0.2 * 38.1±0.1 ** 38.5±0.2
*P<0.05 *P<0.01 * *Compare with model control group p<0.001
Four, analgesic test
6, Herba Senecionis cannabifolii extract causes the influence of pain to the mice hot plate method
Close before each test group mice administration to the threshold value of hot plate method pain reaction, after administration, each dosage group mice of Herba Senecionis cannabifolii extract licks the time of metapedes all than blank group significant prolongation at 1h, 2h, 3h, improve the threshold value of pain reaction, show that Herba Senecionis cannabifolii extract has significant analgesic activity, the Herba Senecionis cannabifolii granule also significantly improves the threshold value of mice hot plate method pain reaction.(table 20-6)
Table 20-6 Herba Senecionis cannabifolii extract is to the analgesic activity of mice hot plate method (X ± s)
Group Number of animals (only) Dosage (g/kg) The preceding threshold of pain of administration (s) The threshold of pain after the administration (s)
1h 2h 3h
Dosage group Herba Senecionis cannabifolii extract high dose group Fanhuncao Granule group in the blank group Herba Senecionis cannabifolii extract low dose group Herba Senecionis cannabifolii extract 12 12 12 12 12 - 8 16 32 16 19.15±3.78 19.38±2.96 20.24±2.56 19.76±3.74 19.15±3.46 20.08±2.97 29.16±4.67 ** 35.50±6.54 ** 40.83±4.15 *** 31.08±4.26 ** 20.24±3.16 26.00±2.79 * 32.12±2.95 ** 34.65±3.84 *** 30.35±3.29 ** 20.43±2.85 24.25±3.16 * 30.75±3.15 ** 33.58±2.15 *** 28.16±2.97 **
*P<0.05, *P<0.01 * *Compare with the blank group P<0.001
7, Herba Senecionis cannabifolii extract is to the influence of mice optical heat radiation method pain reaction
It is approaching before each test group mice administration photo-thermal to be caused the threshold of reaction bitterly, after administration, each dosage group mice of Herba Senecionis cannabifolii extract 1h, 2h, 3h whipping time all than the model control group significant prolongation, improve the threshold value of pain reaction, show that Herba Senecionis cannabifolii extract has significant analgesic activity (table 20-7).
Table 20-7 Herba Senecionis cannabifolii extract is to the analgesic activity of mice photo-thermal method (X ± s)
Group Number of animals (only) Dosage (gkg) The administration threshold of pain (s) The threshold of pain after the administration (s)
1h 2h 3h
Dosage group Herba Senecionis cannabifolii extract high dose group Herba Senecionis cannabifolii groups of grains in the blank group Herba Senecionis cannabifolii extract low dose group Herba Senecionis cannabifolii extract 12 12 12 12 12 - 8 16 32 16 6.66±2.78 6.55±2.18 6.58±1.85 6.24±2.05 6.38±1.95 6.25±2.48 9.44±2.13 ** 11.26±2.27 ** 12.18±3.15 *** 10.38±2.75 ** 6.17±2.52 10.52±2.18 ** 10.38±3.10 ** 11.85±2.97 *** 9.37±3.12 ** 6.44±1.98 8.25±2.04 * 10.11±2.08 ** 10.98±2.87 ** 8.98±2.84 **
*P<0.05, *P<0.01 * *Compare with the blank group P<0.001
8, the Herba Senecionis cannabifolii extract Dichlorodiphenyl Acetate causes the influence of mouse writhing experiment
The result shows: Herba Senecionis cannabifolii extract all can significantly suppress the number of times that acetic acid causes mouse writhing under used dosage, and its effect is suitable with the Herba Senecionis cannabifolii granule, and is dose-effect relationship.See Table 20-8.
Table 20-8 Herba Senecionis cannabifolii extract Dichlorodiphenyl Acetate cause the mouse writhing experiment influence (X ± SD, n=12)
Group Dosage (g/kg) Turn round the body number of times Analgesia rate (%)
Model control group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 8 16 32 16 40.1±4.7 21.5±11.0 *** 17.5±7.1 *** 15.9±4.1 *** 19.7±6.9 *** - 46.4 56.4 60.3 50.8
*P<0.05, *Compare with model group p<0.01
Five, immunity test
9, to the influence of mice reticuloendothelial system (RES) phagocytic function
The result shows: Herba Senecionis cannabifolii extract all can significantly strengthen its RES phagocytic function under used dosage, and the K value is all seen and obviously increased and be dose-effect relationship.See Table 20-9.
Table 20-9 Herba Senecionis cannabifolii extract to the influence of mice RES phagocytic function (X ± SD, n=10)
Group Dosage (g/kg) Phagocytic index K The P value
Normal saline group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 8 16 32 16 0.0301±0.0101 0.0409±0.0098 0.0442±0.0065 0.0471±0.0083 0.0423±0.0097 - <0.05 <0.05 <0.01 <0.05
10, the influence that sheep red blood cell (SRBC) induced mice hemolytic antibody is generated
The result shows: Herba Senecionis cannabifolii extract hemolysin under used dosage increases, and the OD value increases, and then explanation has the effect that strengthens humoral immunization.See Table 20-10.
The influence that table 20-10 Herba Senecionis cannabifolii extract generates sheep red blood cell (SRBC) induced mice hemolytic antibody (X ± SD, n=10)
Group Dosage (g/kg) Antibody concentration (OD) value The P value
Normal saline group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 8 16 32 16 0.26±0.059 0.31±0.047 0.35±0.044 0.42±0.056 0.32±0.049 - >0.05 <0.05 <0.01 <0.05
Six, antiallergic test
11, to the influence of mice DCHR
The results are shown in Table 20-11.By the result as can be known, each dosage group of Herba Senecionis cannabifolii extract all can significantly suppress the mice DCHR, significantly suppresses mice ear, and high dose group increases mouse spleen index and thymus index in the Herba Senecionis cannabifolii extract.
Table 20-11 Herba Senecionis cannabifolii extract to the influence of mice DCHR (X ± SD, n=10)
Group Dosage (g/kg) Ear swelling degree (mg) Spleen index (mg/10g) Thymus index (mg/10g)
Blank group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 32 16 8 16 18.8±2.65 10.5±3.15 ** 11.6±2.97 ** 15.7±3.23 13.7±2.45 * 52.1±11.2 68.0±11.0 ** 63.2±7.3 * 60.1±10.5 62.9±8.2 * 23.1±3.0 31.9±2.9 * 29.2±3.0 * 26.8±3.2 28.7±2.1 *
*P<0.05, *Compare with the blank group p<0.01
Seven, cough-relieving, relieving asthma, expectorant test
12, the Cavia porcellus medicine is drawn the protective effect of breathing heavily
The results are shown in Table 20-12.By the result as can be known, but Herba Senecionis cannabifolii extract and Herba Senecionis cannabifolii granule all significant prolongation cause the incubation period that Cavia porcellus pants by the spraying of acetylcholine and histamine mixed liquor.
Table 20-12 Herba Senecionis cannabifolii extract to the Cavia porcellus medicine draw the protective effect of breathing heavily influence (X ± SD, n=8)
Group Dosage (g/kg) The breathing heavily time (s) Protective rate (%)
Model control group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 4.8 9.5 19.0 9.5 59.6±15.8 160.5±115.8 * 222.6±137.4 * 266.4±121.6 ** 193.4±138.8 * - 25 50 62.5 37.5
*P<0.05, *Compare with model group p<0.01
13, to the influence of phenol red output of mouse breathing road
The result shows: Herba Senecionis cannabifolii extract has tangible phlegm-dispelling functions under used dosage, and is dose-effect relationship.See Table 20-13
Table 20-13 Herba Senecionis cannabifolii extract to the influence of phenol red output of mouse breathing road (X ± SD, n=10)
Group Dosage (g/kg) Phenol red output (OD), be worth
Normal saline group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 8 16 32 16 0.184±0.111 0.209±0.055 * 0.266±0.027 ** 0.302±0.053 *** 0.255±0.037 **
*P<0.05, *P<0.01, * *Compare with the normal saline group p<0.001
14, mice ammonia is caused the influence of coughing
The result: Herba Senecionis cannabifolii extract has significant antitussive effect in each dosage group.See Table 20-14
Table 20-14 injection Herba Senecionis cannabifolii extract to mice ammonia cause the influence coughed (X ± SD, n=10)
Group Dosage (mg/kg) Cough number of times (in the 2min) Suppression ratio
Model group Herba Senecionis cannabifolii extract Herba Senecionis cannabifolii granule - 32 16 8 16 41.1±17.1 19.3±11.8 *** 24.5±14.6 ** 31.6±17.6 * 26.6±14.7 * - 53.4 40.4 23.1 35.4
*P<0.05, *P<0.01, * *Compare with model group p<0.001
In sum, Herba Senecionis cannabifolii extract of the present invention has antiviral significantly, antibiotic, analgesic, antiinflammatory, analgesia cough-relieving, relieving asthma, eliminate the phlegm and the effect of enhance immunity, and certain anti-allergic effects is arranged, so the present invention also comprises with the application in the medicine of the above-mentioned disease of extract preparation treatment of the present invention.Its function is that lung qi dispersing is eliminating evil with curing mainly, heat-clearing and toxic substances removing, antitussive and antiasthmatic.Be used for upper respiratory tract infection, acute tracheobronchitis, acute episode of chronic bronchitis wind heat excess syndrome person, disease is seen heating, pharyngalgia, cough, expectoration, the turbid tears of stream or nasal obstruction, is convenient to yellowish urine, red tongue with yellow fur, floating pulse, sliding number etc.
The specific embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
Embodiment 1: the preparation of extract
Get the 5000g Herba Senecionis cannabifolii, decoct with water three times, add 10 times in water respectively, 8 times, 8 times, each 1 hour, collecting decoction filtered, filtrate is concentrated into the clear paste that relative density is 1.15~1.25g/ml (80 ℃ of survey), puts coldly, adds 95% ethanol, mixing, to pure relative density 75%, left standstill precipitation 12 hours, filter, decompression filtrate recycling ethanol, being concentrated into relative density is the clear paste of 1.10~1.20g/ml (60 ℃ of surveys), puts cold, with clear paste: the amount of resin=1: 1, last cation exchange resin column adsorbs 30min on resin column, then the water eluting, collect water elution part spray drying, promptly get extract 70g.
Embodiment 2: the preparation of extract
Get the 5000g Herba Senecionis cannabifolii, decoct with water three times, add 12 times in water respectively, 10 times, 10 times, each 2 hours, collecting decoction filtered, filtrate is concentrated into the clear paste that relative density is 1.15~1.25g/ml (80 ℃ of survey), puts coldly, adds 95% ethanol, mixing, to pure relative density 75%, left standstill precipitation 24 hours, filter, decompression filtrate recycling ethanol, being concentrated into relative density is the clear paste of 1.10~1.20g/ml (60 ℃ of surveys), puts cold, with clear paste: the amount of resin=1: 1, last cation exchange resin column adsorbs 60min on resin column, then the water eluting, collect water elution part spray drying, promptly get extract 75g.
Embodiment 3: the preparation of extract
Get the 5000g Herba Senecionis cannabifolii, decoct with water three times, add 9 times in water respectively, 8 times, 8 times, each 3 hours, collecting decoction filtered, filtrate is concentrated into the clear paste that relative density is 1.15~1.25g/ml (80 ℃ of survey), puts coldly, adds 85% ethanol, mixing, to pure relative density 75%, left standstill precipitation 6 hours, filter, decompression filtrate recycling ethanol, being concentrated into relative density is the clear paste of 1.10~1.20g/ml (60 ℃ of surveys), puts cold, with clear paste: the amount of resin=1: 1, last cation exchange resin column adsorbs 60min on resin column, then the water eluting, collect water elution part spray drying, promptly get extract 65g.
Embodiment 4 capsule preparations
Extract among the embodiment 1 is pulverized and is granulated, and adds micropowder silica gel and the dextrin composite auxiliary material by 4: 1 compositions, magnesium carbonate, and mixing promptly can be made into capsule by the capsule method for making.
Embodiment 5
Tablet formulation is as follows:
The extract 70g of embodiment 1
Starch 60g
Lactose 30g
Hydroxypropyl cellulose 1.25g
Magnesium stearate 0.5g
Pulvis Talci 5g
Make 1000
Technology
Get extract 60g, add starch 60g, lactose 30g, hydroxypropyl cellulose 1.25g, make granule, drying, granulate adds Pulvis Talci 5g and magnesium stearate 0.5g mix homogeneously, makes 1000, sugar coating, promptly.
Embodiment 6
The granule prescription is as follows:
The extract 60g of embodiment 1
Starch 60g
Sucrose 30g
Hydroxypropyl cellulose 1.25g
Make 1000g
Preparation process
Get the extract 60g of embodiment 1, add starch 60g, sucrose 30g, hydroxypropyl cellulose 1.25g, make granule, drying, granulate, pack 1000g is promptly.
Embodiment 7
The pellet prescription is as follows:
The extract 60g of embodiment 1
Starch 60g
Sucrose 30g
Hydroxypropyl cellulose 1.25g
Make 1000g
Preparation process
Get the extract 60g of embodiment 1, add starch 60g, sucrose 30g, hydroxypropyl cellulose 1.25g, be put in the micropill machine, make micropill, drying, granulate, pack 1000g is promptly.

Claims (10)

1, a kind of Herba Senecionis cannabifolii extract is characterized in that, does not contain or contain hardly alkaloid component in the described extract.
2, the extract of claim 1 is characterized in that, alkaloid is less than 0.5% in the described extract.
3, the extract of claim 1 is characterized in that, described extract is carried through water, precipitate with ethanol, last cation exchange resin column step preparation.
4, the extract of claim 3 is characterized in that, wherein said water is carried, and is that process decocts with water 1-4 time, and the amount that at every turn adds entry is 5-15 times, each 1-3 hour; Described precipitate with ethanol, be with water extract concentrate obtain clear paste after, add the ethanol of 50-100%, left standstill 6-24 hour, treat that precipitation separates out after-filtration and fall precipitation, reclaim ethanol, concentrate and obtain clear paste; Described upward cation exchange resin column is that the clear paste that will obtain behind the precipitate with ethanol joins cation exchange resin column, adsorbs 10-60 minute, and the water eluting obtains water elution liquid then, and this eluent drying obtains extract.
5, the extract of claim 4 is characterized in that, wherein said resin is the polystyrene cation exchange resin, or acrylic acid type cation exchange resin, and its consumption is a clear paste: resin=1: 2-2: 1.
6, the preparation method of extract of claim 1 is characterized in that, puies forward precipitate with ethanol, last cation exchange resin column step through water.
7, the preparation method of claim 6 is characterized in that, process following steps: get Herba Senecionis cannabifolii, decoct with water three times, add water 12-9 respectively doubly, 10-8 doubly, 10-8 times, each 1-1.5 hour, collecting decoction, filter, filtrate is concentrated into the clear paste that relative density is 1.10~1.30g/ml (80 ℃ of survey), puts coldly, adds 95% ethanol, mixing, to pure relative density 75%, left standstill precipitation 12 hours, filter, decompression filtrate recycling ethanol, being concentrated into relative density is the clear paste of 1.10~1.20g/ml (60 ℃ of surveys), puts cold, amount with clear paste: resin=1-2: 1-2, last cation exchange resin column adsorbs 20-40min on resin column, then the water eluting, collect water elution part spray drying, promptly get extract.
8, contain the pharmaceutical preparation of the extract of claim 1.
9, the described pharmaceutical preparation of claim 8 is tablet, capsule, oral liquid, syrup, granule, pill, pellet, powder, unguentum, sublimed preparation, injection, suppository, cream, spray, drop pill, patch, slow releasing preparation or controlled release preparation.
10, the extract with claim 1 prepares a kind of treatment upper respiratory tract infection, acute tracheobronchitis, acute episode of chronic bronchitis wind heat excess syndrome person, disease sees heating, pharyngalgia, cough, expectoration, the turbid tears of stream or nasal obstruction, dry stool yellowish urine, red tongue with yellow fur, floating pulse, sliding number disease or antiviral, antibiotic, analgesic, antiinflammatory, analgesia cough-relieving, relieving asthma, eliminate the phlegm and the medicine of enhance immunity in application.
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CN112569268A (en) * 2020-12-27 2021-03-30 吉林修正药业新药开发有限公司 A composition for treating respiratory system diseases and its preparation method
CN115969874A (en) * 2023-03-20 2023-04-18 吉林华康药业股份有限公司 Pharmaceutical composition for resisting influenza virus

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