CN103494915B - Preparation method and application of breast lump dissipating tablet - Google Patents
Preparation method and application of breast lump dissipating tablet Download PDFInfo
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- CN103494915B CN103494915B CN201310457112.5A CN201310457112A CN103494915B CN 103494915 B CN103494915 B CN 103494915B CN 201310457112 A CN201310457112 A CN 201310457112A CN 103494915 B CN103494915 B CN 103494915B
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Abstract
The invention provides a preparation method of a breast lump dissipating tablet. The breast lump dissipating tablet is prepared from bulk drugs, including 825g of tangerine leaves, 825g of roots of red-rooted salvia, 550g of spina gleditsiae, 550g of seeds of cowherb, 550g of szechwan chinaberry fruit, and 550g of earthworms, through supercritical extraction, so that the content is improved greatly, and the dose is reduced. The invention also provides an application of the breast lump dissipating tablet in the preparation of medicines for inhibiting cell proliferation of mouse melanoma lung metastatic lines B16-F10.
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 RUKUAIXIAO PIAN.
Background technology
RUKUAIXIAO PIAN is recorded in pharmacopeia, prescription is Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, above Six-element, except Pheretima, Semen Vaccariae, four tastes such as all the other Folium Citri tangerinae decoct with water secondary, each 1 hour, collecting decoction, filter, it is 1.25 ~ 1.30(85 DEG C that filtrate is concentrated into relative density), let cool, for subsequent use; Pheretima, Semen Vaccariae 70% alcohol reflux secondary, 2 hours first times, second time 1 hour, filters, merging filtrate, adds in above-mentioned concentrated solution, and adjustment amount of alcohol reaches 70%, stir, leave standstill, reclaim ethanol and be concentrated into paste, drying under reduced pressure becomes dry extract, pulverizes, adds right amount of auxiliary materials, mixing, makes granule, dry, be pressed into 1000, sugar coating, obtain final product.Energy depressed liver-energy dispersing and QI regulating, blood circulation promoting and blood stasis dispelling, dissolving breast mass.For depression of liver-QI, qi depression to blood stasis, cyclomastopathy, distending pain of the breast.
In prior art, not yet have RUKUAIXIAO PIAN extracting the report adopting supercritical technology in preparation, and adopt the method for soak by water, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, is inconvenient to take, and has had a strong impact on this product and has applied clinically.
In prior art, the every sheet 0.5g of RUKUAIXIAO PIAN, each 8-12 sheet, 2 times on the one, adopt the every sheet 0.5g of RUKUAIXIAO PIAN that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only needs 5 at every turn, within 1st, take 2 times, 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 RUKUAIXIAO PIAN.
Another object of the present invention is to provide RUKUAIXIAO PIAN to suppress the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of RUKUAIXIAO PIAN, be made up as crude drug of Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, described method is made up of the following step: get Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, join 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, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 1000, the heavy 0.5g of every sheet.
The preparation method of above-mentioned a kind of RUKUAIXIAO PIAN, described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
The preparation method of above-mentioned a kind of RUKUAIXIAO PIAN, described CO
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation, RUKUAIXIAO PIAN is made up as crude drug of Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, preparation method is made up of the following step: get Radix Bupleuri, joins 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, adds starch, 70% ethanol granule, and dry, tabletting, makes 1000, the heavy 0.5g of every sheet.
Above-mentioned a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation, CO described in the preparation method of RUKUAIXIAO PIAN
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation, CO described in the preparation method of RUKUAIXIAO PIAN
2the extracting pressure 20MPa of supercritical extraction, temperature 40 DEG C, CO
2flow 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 Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO
2flow 1m1/g crude drug min, extraction time 150min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 2
Get Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extracting pressure 30MPa, temperature 50 C, CO
2flow 3m1/g crude drug min, extraction time 180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 1000, the heavy 0.5g of every sheet.
Embodiment 3
Get Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g join CO
2in supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, temperature 40 DEG C, CO
2flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 1000, the heavy 0.5g of every sheet.Embodiment 4: RUKUAIXIAO PIAN suppresses the experimentation data of B16-F10 cell proliferation
1 experiment material
1.1 experiment cell strains
Murine melanoma Lung metastases strain (B16-F10), Nanjing Zhengliang Pharmaceutical Technology Co., Ltd.'s laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: RUKUAIXIAO PIAN of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg RUKUAIXIAO PIAN, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ ldoff pipe subpackages ,-20 DEG C of storages, and simultaneously 0.2 μm of frit dehydrated alcohol is in order to the use of matched group.
1.3 experiment reagent
DMEM(GIBCO company Cat.No.12100-061Lot.No.758137); Hyclone (Tian Hang bio tech ltd, Zhejiang Lot.No.100419); NaHCO3(Shanghai hundred million chemical reagent company limited Cat.No.11810-033Lot.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 equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (MD company of U.S. model: SPECTRA MAX190); 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/N020579); 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) B16-F10 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 3ml0.25% 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 RUKUAIXIAO PIAN 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 statistical dispositions
Adopt the correlation analysis in Microsoft Excel2003 software and Student t to check, data represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, compare with matched group, when dosage reaches 5mg/ml, to B16-F10 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 RUKUAIXIAO PIAN is to B16-F10 cell inhibitory effect influence research (X ± SD)
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
RUKUAIXIAO PIAN can suppress B16-F10 cell proliferation, and reduce the Growth of Cells number of B16-F10 cell, this effect is dose dependent.
Claims (3)
1. the application of RUKUAIXIAO PIAN in preparation suppression murine melanoma Lung metastases strain B16-F10 cell proliferation, it is characterized in that RUKUAIXIAO PIAN is made up as crude drug of Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, preparation method is made up of the following step: get Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, 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, adds starch, 70% ethanol granule, and dry, tabletting, makes 1000, the heavy 0.5g of every sheet.
2. a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation according to claim 1, CO described in the preparation method that it is characterized in that RUKUAIXIAO PIAN
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma Lung metastases strain B16-F10 cell proliferation in preparation according to claim 1, CO described in the preparation method that it is characterized in that RUKUAIXIAO PIAN
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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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107441150A (en) * | 2017-06-28 | 2017-12-08 | 江苏省徐州医药高等职业学校 | The technique of general flavone in a kind of spina gleditsiae using supercritical carbon dioxide extraction |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106511915B (en) * | 2017-01-16 | 2019-06-21 | 江苏省中医院 | A kind of compound traditional Chinese medicine composite is inhibiting the application in cutaneous melanoma Lung metastases drug |
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Non-Patent Citations (2)
| Title |
|---|
| 国家药典委员会.乳块消片.《中华人民共和国药典一部》.2005,第500-501页. * |
| 廖传华等.超临界CO2流体萃取技术——工艺开发及其应用.《超临界CO2流体萃取技术——工艺开发及其应用》.2004,第245-249页. * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107441150A (en) * | 2017-06-28 | 2017-12-08 | 江苏省徐州医药高等职业学校 | The technique of general flavone in a kind of spina gleditsiae using supercritical carbon dioxide extraction |
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