CN103494915A - 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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- CN103494915A CN103494915A CN201310457112.5A CN201310457112A CN103494915A CN 103494915 A CN103494915 A CN 103494915A CN 201310457112 A CN201310457112 A CN 201310457112A CN 103494915 A CN103494915 A CN 103494915A
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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 the Chinese medicine preparation technical field, 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 flavors such as all the other Folium Citri tangerinae decoct with water secondary, each 1 hour, collecting decoction, filter, and it is 1.25～1.30(85 ℃ that filtrate is concentrated into relative density), let cool, standby; Pheretima, 70% alcohol reflux secondary for Semen Vaccariae, 2 hours for the first time, 1 hour for the second time, filter, merging filtrate, add in above-mentioned concentrated solution, adjusts amount of alcohol and reach 70%, stir, standing, reclaim ethanol and be concentrated into the thick paste shape, drying under reduced pressure becomes dry extract, pulverizes, and adds right amount of auxiliary materials, mix granulation, drying, be pressed into 1000, sugar coating, obtain.The 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 there is RUKUAIXIAO PIAN adopting the report of supercritical technology aspect the extraction preparation, and the method that adopts decocting to boil, 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.
Summary of the invention
Goal of the invention: in order to address the above 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 transfer strain B16-F10 cell proliferation medicine in preparation.
Technical scheme: the objective of the invention is to realize by following scheme:
A kind of preparation method of RUKUAIXIAO PIAN, by Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, as crude drug, made, described method is comprised 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 ℃, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1000, every heavy 0.5g.
The preparation method of above-mentioned a kind of RUKUAIXIAO PIAN, described CO
2the percent by volume that the 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, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma lung transfer strain B16-F10 cell proliferation medicine in preparation, RUKUAIXIAO PIAN is made as crude drug by Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, preparation method is comprised of the following step: get Radix Bupleuri, join CO
2in the 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 ℃, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1000, every heavy 0.5g.
Above-mentioned a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma lung transfer strain B16-F10 cell proliferation medicine, CO described in the preparation method of RUKUAIXIAO PIAN in preparation
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma lung transfer strain B16-F10 cell proliferation medicine, CO described in the preparation method of RUKUAIXIAO PIAN in preparation
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
In prior art, every 0.5g of RUKUAIXIAO PIAN, each 8-12 sheet, 2 times on the one, the every 0.5g of RUKUAIXIAO PIAN that adopts the present invention to be prepared into, but the medical material amount contained is original 2 times, therefore only needs 5 at every turn, within 1st, take 2 times, greatly reduced dose having under the condition of more active component.
The specific embodiment
Form by the following examples, foregoing of the present invention is described in further detail again, but this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits 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 the supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, 30 ℃ of temperature, CO
2flow 1m1/g crude drug min, extraction time 150min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 500, every heavy 0.5g.
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 the 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, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1000, every heavy 0.5g.
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 the supercritical extraction device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extracting pressure 20MPa, 40 ℃ of temperature, CO
2flow 2m1/g crude drug min, extraction time 160min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1000, every heavy 0.5g.Embodiment 4: RUKUAIXIAO PIAN suppresses the experimentation data of B16-F10 cell proliferation
1 experiment material
1.1 experiment cell strain
The murine melanoma lung shifts strain (B16-F10), Nanjing Zheng Liang Pharmaceutical Technology Co., Ltd laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: RUKUAIXIAO PIAN of the present invention: press embodiment 3 method preparations.
The medicinal liquid liquid storage: take the 100mg RUKUAIXIAO PIAN, be dissolved in the 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ ldoff pipe packing, and-20 ℃ of storages, 0.2 μ m filter filters the use of dehydrated alcohol in order to matched group simultaneously.
1.3 experiment reagent
The Cat.No.12100-061Lot.No.758137 of DMEM(GIBCO company); Hyclone (Hangzhoupro, sky, Zhejiang bio tech ltd 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); The autogamy of PBS(laboratory); 1.4 experiment equipment
Lycra inverted microscope (German Leica model: DM1L); Visible-ultraviolet light microwell plate detector (U.S. MD company model: SPECTRA MAX190); CO2 incubator (FORMA model: 3111); (safe and sound company of Su Jing group manufactures model to super-clean bench: SW-CJ-ZFD); Pure water instrument (U.S. Spring company 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 (Shanghai accurate experimental facilities company 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 μ m 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) the B16-F10 cell carries out cellar culture (10cm culture dish) with DMEM+10%FBS in 37 ℃, 5%CO2, when Growth of Cells during to logarithmic (log) phase, collecting cell, discard culture fluid, PBS fine laundering 3 times, add 3ml0.25% trypsin-0.04%EDTA, after 37 ℃ of digestion 2min, add wherein 5ml complete medium neutralization reaction, after the piping and druming cell, it is proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjust 3 * 104/ml of concentration of cell suspension.
2) the cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, culture plate put into cell culture incubator (37 ℃, 5%CO2) cellar culture.
3) according to the Growth of Cells situation, 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, the buckle method is removed supernatant, with absorbent paper, pats dry gently, and every hole adds 200 μ l dimethyl sulfoxide, puts low-speed oscillation 10min on shaking table, and crystal is fully dissolved.Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place.6) background (do not add cell, only add culture fluid) is set simultaneously, control wells (the medicine dissolution medium of cell, same concentrations, culture fluid, MTT, dimethyl sulfoxide), set 6 multiple holes for every group.
7) with medicine, the suppression ratio to cell means result:
Cell increment suppression ratio (%)=(control wells OD value-dosing holes OD value)/control wells OD value * 100%.Experiment repeats 3 times.
3 statistical dispositions
Adopt correlation analysis and Student t check in Microsoft Excel2003 software, data mean with mean ± S.D..
4 experimental results
Statistical result showed after the mtt assay experiment, with matched group, compare, 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), and utmost point significant difference (P<0.001) is arranged when dosage reaches 15-20mg/ml.
Table 1 RUKUAIXIAO PIAN is on B16-F10 cell inhibitory effect impact research (X ± SD)
Annotate: compare * P<0.01 with matched group; * P<0.001
5 experiment conclusion
RUKUAIXIAO PIAN can suppress the B16-F10 cell proliferation, reduces the Growth of Cells number of B16-F10 cell, and this effect is dose dependent.
Claims (6)
1. the preparation method of a RUKUAIXIAO PIAN, by Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, as crude drug, made, it is characterized in that described method is comprised of the following step: get Radix Bupleuri, join in CO2 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 ℃, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1000, every heavy 0.5g.
2. a kind of preparation method of RUKUAIXIAO PIAN according to claim 1, is characterized in that described CO
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of preparation method of RUKUAIXIAO PIAN according to claim 1, is characterized in that described CO
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
4. a kind of RUKUAIXIAO PIAN suppresses the application in murine melanoma lung transfer strain B16-F10 cell proliferation medicine in preparation according to claim 1, it is characterized in that RUKUAIXIAO PIAN made as crude drug by Folium Citri tangerinae 825g, Radix Salviae Miltiorrhizae 825g, Spina Gleditsiae 550g, Semen Vaccariae 550g, Fructus Toosendan 550g, Pheretima 550g, preparation method is comprised of the following step: get Radix Bupleuri, join CO
2in the 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 ℃, CO
2flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, adds starch, 70% ethanol granule processed, drying, tabletting, make 1000, every heavy 0.5g.
5. a kind of RUKUAIXIAO PIAN suppresses the application of murine melanoma lung in shifting strain B16-F10 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in the preparation method of RUKUAIXIAO PIAN
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of RUKUAIXIAO PIAN suppresses the application of murine melanoma lung in shifting strain B16-F10 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in the preparation method of RUKUAIXIAO PIAN
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106511915A (en) * | 2017-01-16 | 2017-03-22 | 江苏省中医院 | Applications of compound traditional Chinese medicinal composition in preparing medicines for inhibiting skin melanoma lung metastasis |
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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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Non-Patent Citations (2)
| Title |
|---|
| 国家药典委员会: "《中华人民共和国药典一部》", 31 January 2005 * |
| 廖传华等: "《超临界CO2流体萃取技术——工艺开发及其应用》", 31 July 2004 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106511915A (en) * | 2017-01-16 | 2017-03-22 | 江苏省中医院 | Applications of compound traditional Chinese medicinal composition in preparing medicines for inhibiting skin melanoma lung metastasis |
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