CN103494878A - Preparation method and application of compound danshen tablets - Google Patents
Preparation method and application of compound danshen tablets Download PDFInfo
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- CN103494878A CN103494878A CN201310465801.0A CN201310465801A CN103494878A CN 103494878 A CN103494878 A CN 103494878A CN 201310465801 A CN201310465801 A CN 201310465801A CN 103494878 A CN103494878 A CN 103494878A
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Abstract
The invention provides a preparation method of compound danshen tablets. The compound danshen tablets are prepared from 450g of danshen, 141g of pseudo-ginseng and 8g of borneol which are used as raw medicinal materials by adopting a supercritical extraction method, so that the content is greatly increased and the intake dosage is reduced. The invention also provides an application of the compound danshen tablets in preparation of a medicament for restricting cell proliferation of mouse melanoma cells B16.
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 FUFANG DANSHEN PIAN.
Background technology
FUFANG DANSHEN PIAN is recorded in pharmacopeia, writes out a prescription as Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, above three flavors, and Radix Salviae Miltiorrhizae extracts three times, adds for the first time alcohol reflux 1.5 hours, filters, filtrate recycling ethanol, be concentrated into relative density 1.30(55～60 ℃); Add for the second time 50% alcohol reflux 1.5 hours, filter; Add for the third time water and reflux 2 hours, filter, merge filtrate second and third time, reclaim ethanol, be concentrated into relative density 1.40(55～60 ℃), with primary concentrated solution, merge, mix, making relative density is the clear paste of 1.35～1.39 (55 ℃).Radix Notoginseng powder is broken into to fine powder, mixes thoroughly with the Radix Salviae Miltiorrhizae clear paste, drying, granulation, by the Borneolum Syntheticum porphyrize, mix with above-mentioned granule, is pressed into 1000, or sugar coating or film-coat, obtains.
In prior art, not yet there is FUFANG DANSHEN PIAN to adopt the report of supercritical technology extracting aspect preparation, and adopt the method that powder and decocting boil of beating, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, be inconvenient to take, had a strong impact on this product and 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 FUFANG DANSHEN PIAN.
Another object of the present invention is to provide a kind of FUFANG DANSHEN PIAN to suppress the application in B16 mouse melanoma cell line cell proliferation medicine in preparation.
Technical scheme: the objective of the invention is to realize by following scheme:
A kind of preparation method of FUFANG DANSHEN PIAN, by Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, as crude drug, made, described method is comprised of the following step: get Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, 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.
The preparation method of above-mentioned a kind of FUFANG DANSHEN 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 FUFANG DANSHEN 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 FUFANG DANSHEN PIAN suppresses the application in B16 mouse melanoma cell line cell proliferation medicine in preparation, FUFANG DANSHEN PIAN is made as crude drug by Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, preparation method is comprised of the following step: get Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, 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, and 70%
ethanolgranule processed, drying, tabletting, make 500, every heavy 0.5g.
Above-mentioned FUFANG DANSHEN PIAN suppresses the application in B16 mouse melanoma cell line cell proliferation medicine, CO described in the compound Salviae Miltiorrhizae piece preparation method in preparation
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned FUFANG DANSHEN PIAN suppresses the application in B16 mouse melanoma cell line cell proliferation medicine, CO described in the compound Salviae Miltiorrhizae piece preparation method 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 FUFANG DANSHEN PIAN, each 8-10 sheet, 2 times on the one, the every 0.5g of FUFANG DANSHEN PIAN that adopts the present invention to be prepared into, but the medical material amount contained is original 2 times, therefore only needs 3 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 Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g joins 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 Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g joins 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 500, every heavy 0.5g.
Embodiment 3
Get Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g joins 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 500, every heavy 0.5g.
Embodiment 4: FUFANG DANSHEN PIAN suppresses the experimentation data of B16 cell proliferation
1 experiment material
1.1 experiment cell strain
Mouse melanin tumor cell (B16), Nanjing Zheng Liang Pharmaceutical Technology Co., Ltd laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: FUFANG DANSHEN PIAN of the present invention: press embodiment 3 method preparations.
The medicinal liquid liquid storage: take the 100mg FUFANG DANSHEN 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 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 FUFANG DANSHEN 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 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 FUFANG DANSHEN PIAN is on B16 cell inhibitory effect impact research
Annotate: with matched group, compare,
*p<0.01;
*p<0.001
5 experiment conclusion
FUFANG DANSHEN PIAN can suppress the B16 cell proliferation, reduces the Growth of Cells number of B16 cell, and this effect is dose dependent.
Claims (6)
1. the preparation method of a FUFANG DANSHEN PIAN, by Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, as crude drug, made, it is characterized in that described method is comprised of the following step: get Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, 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 500, every heavy 0.5g.
2. a kind of preparation method of FUFANG DANSHEN 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 FUFANG DANSHEN 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 FUFANG DANSHEN PIAN suppresses the application in B16 mouse melanoma cell line cell proliferation medicine in preparation according to claim 1, it is characterized in that FUFANG DANSHEN PIAN made as crude drug by Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, preparation method is comprised of the following step: get Radix Salviae Miltiorrhizae 450g, Radix Notoginseng 141g, Borneolum Syntheticum 8g, 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 500, every heavy 0.5g.
5. a kind of FUFANG DANSHEN PIAN suppresses the application in B16 mouse melanoma cell line cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in the compound Salviae Miltiorrhizae piece preparation method
2the percent by volume that the supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of FUFANG DANSHEN PIAN suppresses the application in B16 mouse melanoma cell line cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in the compound Salviae Miltiorrhizae piece preparation method
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
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| CN105395581A (en) * | 2015-12-24 | 2016-03-16 | 济南新时代医药科技有限公司 | Application of folium chimonanthi nitentis tablet to preparation of drug for inhibiting skin melanophore A375 cell proliferation |
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| CN105663719A (en) * | 2016-02-17 | 2016-06-15 | 济南新时代医药科技有限公司 | Application of cough and asthma treating Bulbophyllum calodictyon Schlecht tablets in preparation of medicine for inhibiting proliferation of histocyte lymphoma cells U937 |
| CN105535514A (en) * | 2016-02-17 | 2016-05-04 | 济南新时代医药科技有限公司 | Application of Lvji cough asthma tablets to preparation of medicine for inhibiting cell proliferation of lung adenocarcinma cells H1299 |
| CN105521298A (en) * | 2016-02-17 | 2016-04-27 | 济南新时代医药科技有限公司 | Application of cough and asthma treating tablet containing seudobulb of corymbose coelogyne in preparing medicine for inhibiting cell proliferation of human oral epidermoid carcinoma cells KB |
| CN105535513A (en) * | 2016-02-17 | 2016-05-04 | 济南新时代医药科技有限公司 | Application of Lvji cough asthma tablets to preparation of medicine for inhibiting cell proliferation of gastric adenocarcinoma cells BGC-823 |
| CN105535515A (en) * | 2016-02-17 | 2016-05-04 | 济南新时代医药科技有限公司 | Application of Lvji cough asthma tablets to preparation of medicine for inhibiting cell proliferation of prostate cancer cells PC-3 |
| CN105726960A (en) * | 2016-02-17 | 2016-07-06 | 济南新时代医药科技有限公司 | Application of pseudobulb-of-corymbose-coelogyne cough and asthma tablets to preparation of medicine for restraining human osteosarcoma cell MG-63 multiplication |
| CN105521297A (en) * | 2016-02-17 | 2016-04-27 | 济南新时代医药科技有限公司 | Application of cough and asthma treating tablet containing seudobulb of corymbose coelogyne in preparing medicine for inhibiting cell proliferation of human ileocecal carcinoma cells HCT-8 |
| CN105616995A (en) * | 2016-02-17 | 2016-06-01 | 济南新时代医药科技有限公司 | Application of bulbophyllum and rhizoma bletillae cough and asthma tablets to preparing uterine high-differentiation squamous cancer cell HCC (hepatic cellular cancer) proliferation suppressing medicines |
| CN105616996A (en) * | 2016-02-17 | 2016-06-01 | 济南新时代医药科技有限公司 | Application of bulbophyllum and rhizoma bletillae cough and asthma tablets to preparing embryoma cell F9 proliferation suppressing medicines |
| CN105641447A (en) * | 2016-02-17 | 2016-06-08 | 济南新时代医药科技有限公司 | Application of antitussive and antiasthmatic tablets containing bulbophyllum in preparation of drugs for inhibiting proliferation of human bladder transitional cell carcinoma cells T-24 |
| CN105641446A (en) * | 2016-02-17 | 2016-06-08 | 济南新时代医药科技有限公司 | Application of antitussive and antiasthmatic tablets containing bulbophyllum in preparation of drugs for inhibiting proliferation of human tongue cancer cells TCA 8113 |
| CN105641448A (en) * | 2016-02-17 | 2016-06-08 | 济南新时代医药科技有限公司 | Application of antitussive and antiasthmatic tablets containing bulbophyllum in preparation of drugs for inhibiting proliferation of cervical cancer cells U14 |
| CN105641449A (en) * | 2016-02-17 | 2016-06-08 | 济南新时代医药科技有限公司 | Application of antitussive and antiasthmatic tablets containing bulbophyllum in preparation of drugs for inhibiting proliferation of melanoma cells MM96L |
| CN105663715A (en) * | 2016-02-17 | 2016-06-15 | 济南新时代医药科技有限公司 | Application of cough and asthma treating Bulbophyllum calodictyon Schlecht tablets in preparation of medicine for inhibiting proliferation of astrocytoma cells U251 |
| CN105663718A (en) * | 2016-02-17 | 2016-06-15 | 济南新时代医药科技有限公司 | Application of cough and asthma treating Bulbophyllum calodictyon Schlecht tablets in preparation of medicine for inhibiting proliferation of glioma cells SHG-44 |
| CN105521296A (en) * | 2016-02-17 | 2016-04-27 | 济南新时代医药科技有限公司 | Application of cough and asthma treating tablet containing seudobulb of corymbose coelogyne in preparing medicine for inhibiting cell proliferation of human choriocarcinoma cells JAR |
| CN105663717A (en) * | 2016-02-17 | 2016-06-15 | 济南新时代医药科技有限公司 | Application of cough and asthma treating Bulbophyllum calodictyon Schlecht tablets in preparation of medicine for inhibiting proliferation of kidney cancer cells ACHN |
| CN105663716A (en) * | 2016-02-17 | 2016-06-15 | 济南新时代医药科技有限公司 | Application of cough and asthma treating Bulbophyllum calodictyon Schlecht tablets in preparation of medicine for inhibiting proliferation of leukemia cells K562 |
| CN105687726A (en) * | 2016-02-17 | 2016-06-22 | 济南新时代医药科技有限公司 | Application of Bulbophyllum calodictyon Schlecht.cough-asthma tablets to preparation of medicine for inhibiting proliferation of breast duct cancer cells T47D |
| CN105687728A (en) * | 2016-02-17 | 2016-06-22 | 济南新时代医药科技有限公司 | Application of antitussive and antiasthmatic tablets containing Bulbophyllum calodictyon to preparation of drugs for inhibiting SCaBER cell proliferation |
| CN105687727A (en) * | 2016-02-17 | 2016-06-22 | 济南新时代医药科技有限公司 | Application of Bulbophyllum calodictyon Schlecht.cough-asthma tablets to preparation of medicine for inhibiting proliferation of small intestine cancer cells HIC |
| CN105726961A (en) * | 2016-02-17 | 2016-07-06 | 济南新时代医药科技有限公司 | Application of pseudobulb-of-corymbose-coelogyne cough and asthma tablets to preparation of medicine for restraining well-differentiated nasopharyngeal carcinoma cell CNE-1 multiplication |
| CN105582051A (en) * | 2016-03-02 | 2016-05-18 | 南京正亮医药科技有限公司 | Preparation method and application of compound salvia miltiorrhiza tablet |
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Application publication date: 20140108 |