CN104906059A - Preparation method and application of anti-inflammatory and antasthmatic tablet - Google Patents
Preparation method and application of anti-inflammatory and antasthmatic tablet Download PDFInfo
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- CN104906059A CN104906059A CN201510347459.3A CN201510347459A CN104906059A CN 104906059 A CN104906059 A CN 104906059A CN 201510347459 A CN201510347459 A CN 201510347459A CN 104906059 A CN104906059 A CN 104906059A
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Abstract
The invention belongs to the field of traditional Chinese medicine preparations, and particularly provides a preparation method of an anti-inflammatory and antasthmatic tablet. The anti-inflammatory and antasthmatic tablet is prepared from the following raw materials by weight: 200g of thorny elaeagnus leaf, 150g of platycodon grandiflorum, 200g of radix pseudostellariae, 100g of radix stemonae, 12.5g of pericarpium papaveris, 25g of ephedra, 125g of negundo chastetree fruit, 37.5g of adenophora tetraphylla and 125g of common andrographis herb, and is prepared by a supercritical extraction method, so that the content is greatly improved, and the dosage is reduced. The invention further provides an application of the anti-inflammatory and antasthmatic tablet in preparation of medicines for inhibiting human bladder cancer cell EJ proliferation.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicine preparation, be specifically related to a kind of preparation method and application of inflammation-diminishing and cough-controlling sheet.
Background technology
Inflammation-diminishing and cough-controlling sheet is recorded in national drug food Surveillance Authority standard, is made up as crude drug of Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g.Above nine tastes, Radix Pseudostellariae, Radix Platycodonis, the Radix Stemonae decoct with water three times, 4 hours first times, second time 3 hours, 2 hours third times; Collecting decoction, filters; Folium Elaeagni decocts with water 4 hours, filters, merges with above-mentioned filtrate, is concentrated into the clear paste that relative density is 1.24 (heat is surveyed); The ground spices such as all the other Herba Ephedraes are broken into fine powder, sieve, and mixing, mixes with above-mentioned clear paste, stir evenly, make granule, dry, are pressed into 1200, sugar coating, obtain final product.Antiinflammatory, antitussive, reduces phlegm, Dingchuan.For cough with copious phlegm, fullness in the chest QI rising in reverse order, tracheitis.
In prior art, not yet there is inflammation-diminishing and cough-controlling sheet extracting the report adopting supercritical technology in preparation, and adopt the methods such as water extraction, technique is coarse, complicated, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and applied clinically.
In prior art, the every sheet 0.5g of inflammation-diminishing and cough-controlling sheet, oral, one time 2,3 times on the one, adopt the every sheet 0.5g of inflammation-diminishing and cough-controlling sheet that is prepared into of the present invention, but the medical material amount contained is more than originally, under the condition with more active component, greatly reduce dose.
Summary of the invention
Goal of the invention: in order to solve the problem, the object of the present invention is to provide a kind of preparation method of inflammation-diminishing and cough-controlling sheet.
Another object of the present invention is to provide a kind of inflammation-diminishing and cough-controlling sheet to suppress the application in human bladder cancer cell EJ hyperproliferation agent in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of inflammation-diminishing and cough-controlling sheet, by Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g makes as crude drug, described method is made up of the following step: get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material joins in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, pulverize, add starch 20g, granulate, tabletting, every sheet 0.5g.
The preparation method of described inflammation-diminishing and cough-controlling sheet, the percent by volume that described CO2 supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of described inflammation-diminishing and cough-controlling sheet, the extracting pressure 20MPa of described CO2 supercritical extraction, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
Described inflammation-diminishing and cough-controlling sheet suppresses the application in human bladder cancer cell EJ hyperproliferation agent in preparation, inflammation-diminishing and cough-controlling sheet is by Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g makes as crude drug, get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material joins in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, pulverize, add starch 20g, granulate, tabletting, every sheet 0.5g.
Described inflammation-diminishing and cough-controlling sheet suppresses the application in human bladder cancer cell EJ hyperproliferation agent in preparation, and described in the preparation method that it is characterized in that inflammation-diminishing and cough-controlling sheet, CO2 supercritical extraction entrainer accounts for the percent by volume of total extractant is 5%.
Described inflammation-diminishing and cough-controlling sheet suppresses the application in human bladder cancer cell EJ hyperproliferation agent in preparation, the extracting pressure 20MPa of CO2 supercritical extraction described in the preparation method that it is characterized in that inflammation-diminishing and cough-controlling sheet, temperature 40 DEG C, CO2 flow 2ml/g crude drug min, extraction time 160min.
Detailed description of the invention
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following example, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1: get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 30MPa, temperature 30 DEG C, CO2 flow 3m1/g crude drug min, extraction time 150min, obtain supercritical extract, pulverize, add chlorphenamine maleate, starch, granulate, tabletting, every sheet 0.5g.
Embodiment 2: get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 15MPa, temperature 50 C, CO2 flow 1m1/g crude drug min, extraction time 180min, obtain supercritical extract, pulverize, add chlorphenamine maleate, starch, granulate, tabletting, every sheet 0.5g.
Embodiment 3: get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 25MPa, temperature 40 DEG C, CO2 flow 2m1/g crude drug min, extraction time 160min, obtain supercritical extract, pulverize, add chlorphenamine maleate, starch, granulate, tabletting, every sheet 0.5g.
Folium Elaeagni is the blade of Elaeangnaceae plant Fructus Elaeagni.Fructus Viticis Negundo derives from the fruit of Verbenaceae five-leaved chaste tree Vitex negundo L..
Embodiment 4: inflammation-diminishing and cough-controlling sheet suppresses the experimentation data of human bladder cancer cell EJ propagation
1 experiment material
1.1 experiment cell strains: human bladder cancer cell EJ, Nanjing Medical University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents: drugs: inflammation-diminishing and cough-controlling sheet of the present invention: prepare by embodiment 3 method.Medicinal liquid liquid storage: take 100mg inflammation-diminishing and cough-controlling sheet content, be dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ l doff pipe subpackages ,-20 DEG C of storages, simultaneously 0.2 μm of frit dehydrated alcohol is in order to the use of matched group.
1.3 experiment reagents: DMEM (GIBCO company Cat.No.12100-061 Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3 (Shanghai hundred million chemical reagent company limited Cat.No.11810-033 Lot.No.1088387 of a specified duration); Trypsin (AMRESCO company lot number: 2010/04); EDTA (AMRESCO company lot number: 2009/10); Penicillin G Sodium Salt (AMRESCO company lot number: 2010242); Streptomycin Sulfate (AMRESCO company lot number: 2010382); Dehydrated alcohol (Nanjing Chemistry Reagent Co., Ltd.'s lot number: 080310182); MTT (Biosharp lot number: 0793); PBS (laboratory autogamy);
1.4 experiment equipments: Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRA MAX 190); CO2 incubator (FORMA model: 3111); Super-clean bench (safe and sound Inc. of Su Jing group moulding number: SW-CJ-ZFD); Pure water instrument (Spring company of U.S. model: S/N 020579); Accurate pipettor (French Gilson Inc model: P2); Electronic balance (German Sai Duolisi company limited model: BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air dry oven (the accurate experimental facilities company in Shanghai model: DHG9123A); Refrigerator (Siemens Company's model: KG18V21TI); Liquid nitrogen container (CBS model: 2001); Low speed centrifuge (Anting Scientific Instrument Factory, Shanghai's model: KA-1000); 0.2 μm of filter (MILLIPORE model: SLGP033RB); 10cm culture dish (NEST company), 96 well culture plates (NEST company); Cell counting count board; Centrifuge tube, pipet, Tips are some.
2 experimental techniques: 1) EJ cell DMEM+10%FBS in 37 DEG C, 5%CO2 carries out cellar culture (10cm culture dish), when Growth of Cells is to logarithmic (log) phase, collecting cell, discards culture fluid, PBS fine laundering 3 times, add 3ml 0.25% trypsin-0.04%EDTA, after 37 DEG C of digestion 2min, add 5ml complete medium neutralization reaction wherein, proceeded in centrifuge tube after piping and druming cell, the centrifugal 5min of 1000rpm, adjustment concentration of cell suspension 3 × 104/ml.2) enter in 96 well culture plates by cell kind, every hole adds cell suspension 180 μ l, and culture plate puts into cell culture incubator (37 DEG C, 5%CO2) cellar culture.3) according to cell growth status, generally grow to 50%-70%, add inflammation-diminishing and cough-controlling sheet solution, continue to cultivate 24h.4) add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT) after 24h, continue to cultivate 4h.5) after 4h, buckle method removes supernatant, pats dry gently with absorbent paper, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, crystal is fully dissolved.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm place.6) background (do not add cell, only add culture fluid) is set, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide) simultaneously, often organizes the multiple hole of setting 6.7) result represents with the suppression ratio of medicine to cell: cell proliferation suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%.Experiment repetition 3 times.
3 experimental results: statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to EJ cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has pole significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 inflammation-diminishing and cough-controlling sheet is to human bladder cancer cell EJ Proliferation Ability shadow
research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
4 experiment conclusion: inflammation-diminishing and cough-controlling sheet can suppress human bladder cancer cell EJ to breed, reduce the Growth of Cells number of human bladder cancer cell EJ, this effect is dose dependent.
Claims (6)
1. the preparation method of an inflammation-diminishing and cough-controlling sheet, be made up as crude drug of Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g, it is characterized in that described method is made up of the following step: get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, pulverizes, adds starch 20g, granulates, tabletting, every sheet 0.5g.
2. the preparation method of inflammation-diminishing and cough-controlling sheet according to claim 1, is characterized in that described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. the preparation method of inflammation-diminishing and cough-controlling sheet according to claim 1, is characterized in that described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
4. inflammation-diminishing and cough-controlling sheet suppresses the application in human bladder cancer cell EJ hyperproliferation agent in preparation according to claim 1, it is characterized in that inflammation-diminishing and cough-controlling sheet is by Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g makes as crude drug, preparation method is: get Folium Elaeagni 200g, Radix Platycodonis 150g, Radix Pseudostellariae 200g, Radix Stemonae 100g, Pericarpium Papaveris 12.5g, Herba Ephedrae 25g, Fructus Viticis Negundo 125g, Radix Adenophorae 37.5g, Herba Andrographis 125g medical material joins in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, pulverize, add starch 20g, granulate, tabletting, every sheet 0.5g.
5. inflammation-diminishing and cough-controlling sheet suppresses the application in human bladder cancer cell EJ hyperproliferation agent in preparation according to claim 4, CO described in the preparation method that it is characterized in that inflammation-diminishing and cough-controlling sheet
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. inflammation-diminishing and cough-controlling sheet suppresses the application in human bladder cancer cell EJ hyperproliferation agent in preparation according to claim 4, CO described in the preparation method that it is characterized in that inflammation-diminishing and cough-controlling sheet
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
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