CN103720799B - A kind of preparation method of active pain relieving tablet and application - Google Patents
A kind of preparation method of active pain relieving tablet and application Download PDFInfo
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- CN103720799B CN103720799B CN201310612517.1A CN201310612517A CN103720799B CN 103720799 B CN103720799 B CN 103720799B CN 201310612517 A CN201310612517 A CN 201310612517A CN 103720799 B CN103720799 B CN 103720799B
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Abstract
The invention provides a kind of preparation method of active pain relieving tablet, by wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g, red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g, above ten tastes, get Radix Angelicae Sinensis 120g, red sage root 120g is as bulk drug, employing supercritical extract is prepared from, content is improved a lot, dose reduces, and the present invention also provides active pain relieving tablet to suppress the application in rat neurogliocytoma cells C6 cell proliferation medicine in preparation.
Description
Technical field
The present invention relates to Chinese medicine preparation technical field, be specifically related to a kind of preparation method and application of active pain relieving tablet.
Background technology
Active pain relieving tablet is recorded in the standard WS3-B-0355-90 of the Ministry of Public Health, write out a prescription into wilsonii 100g, root of Chinese clematis 100g,Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g, red sage root 150g, frankincense 15g, myrrh 15g,Chinese ephedra 30g, above ten tastes, get Radix Angelicae Sinensis 120g, red sage root 120g, the root of Chinese clematis and Chinese ephedra, boiling secondary, each 4Hour, collecting decoction, filters, and filtrate is concentrated into the clear cream that relative density is 1.28～1.30 (80 DEG C); Separately get river processedCrow, wild aconite root, frankincense, myrrh, Radde Anemone Rhizome and remaining Radix Angelicae Sinensis, danshen powder are broken into fine powder, sieve, with above-mentioned clearCream, the each right amount of auxiliary materials of Extractum Acanthopanacis Senticosi fully mix, and granulate, dry, are pressed into 1000, and sugar coating, to obtain final product.Can be clearing and activating the channels and collaterals, relaxing muscles and tendons pain relieving. For wind-cold-dampness arthralgia, channels and collaterals obturation, arthralgia and myalgia, numb limb.
In prior art, not yet there is active pain relieving tablet adopting the report of supercritical technology aspect extraction preparation, and adoptBeat the method that powder and decocting boil, technique is coarse, backward, and impurity is many, causes patient's consumption excessive, is inconvenient to take, tightGhost image has rung 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 active pain relieving tablet.
Another object of the present invention is to provide a kind of active pain relieving tablet to suppress rat neurogliocytoma cells C6 in preparationApplication in cell proliferation medicine.
Technical scheme: the object of the invention is to realize by following scheme:
A preparation method for active pain relieving tablet, by wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g,Wild aconite root 40g, Radde Anemone Rhizome 20g, red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g, above ten tastes, getRadix Angelicae Sinensis 120g, red sage root 120g joins CO2In supercritical extract device, ethanol is as entrainer, and entrainer accounts for total extractionThe percent by volume of solvent is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2Flow 1-3ml/g is rawMedicine min, extraction time 150-180min, obtains supercritical extract, adds starch, and 70% ethanol particle processed is dry,Compressing tablet, makes 500, every heavy 0.35g.
The preparation method of above-mentioned a kind of active pain relieving tablet, described CO2Supercritical extract entrainer accounts for the body of total extractantLong-pending percentage is 5%.
The preparation method of above-mentioned a kind of active pain relieving tablet, described CO2The extracting pressure 20MPa of supercritical extract, temperature40℃，CO2Flow 2ml/g crude drug min, extraction time 160min.
Above-mentioned a kind of active pain relieving tablet suppresses the application in rat neurogliocytoma cells C6 cell proliferation medicine in preparation,Active pain relieving tablet is by wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, ring perfume (or spice)Attached 20g, red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g, above ten tastes, get Radix Angelicae Sinensis 120g, red sage root 120gMake as bulk drug, preparation method is made up of the following step: get wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g,Aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g, red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g,Above ten tastes, get Radix Angelicae Sinensis 120g, and red sage root 120g joins CO2In supercritical extract device, ethanol, as entrainer, presss from both sidesThe percent by volume that accounts for total extractant with agent is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2StreamAmount 1-3ml/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol systemParticle, dry, compressing tablet, makes 500, every heavy 0.35g.
Above-mentioned a kind of active pain relieving tablet suppresses the application in rat neurogliocytoma cells C6 cell proliferation medicine in preparation,CO described in the preparation method of active pain relieving tablet2The percent by volume that supercritical extract entrainer accounts for total extractant is 5%.
Above-mentioned a kind of active pain relieving tablet suppresses the application in rat neurogliocytoma cells C6 cell proliferation medicine in preparation,CO described in the preparation method of active pain relieving tablet2The extracting pressure 20MPa of supercritical extract, 40 DEG C of temperature, CO2Flow2ml/g crude drug min, extraction time 160min.
In prior art, every 0.35g of active pain relieving tablet,, 3 times on the one, adopts the present invention to be prepared into by each 4Every 0.35g of active pain relieving tablet, but the medicinal material amount containing is original 2 times, therefore only needs 2 at every turn, clothes on the oneWith 2 times, under the condition with more active components, greatly reduce dose.
Detailed description of the invention
Form by the following examples, is described in further detail foregoing of the present invention again, but should be by thisThe scope that is interpreted as the above-mentioned theme of the present invention only limits to following example, all technology realizing based on foregoing of the present inventionAll belong to scope of the present invention.
Embodiment 1
Get wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g,Red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g, above ten tastes, get Radix Angelicae Sinensis 120g, and red sage root 120g joinsCO2In supercritical extract device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 4%, extractionPressure 15MPa, 30 DEG C of temperature, CO2Flow 1ml/g crude drug min, extraction time 150min, obtains overcritical extractionGet thing, add starch, 70% ethanol particle processed, dry, compressing tablet, makes 500, every heavy 0.35g.
Embodiment 2
Get wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g,Red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g, above ten tastes, get Radix Angelicae Sinensis 120g, and red sage root 120g joins CO2In supercritical extract device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 6%, extractionPressure power 30MPa, temperature 50 C, CO2Flow 3ml/g crude drug min, extraction time 180min, obtains overcriticalExtract, adds starch, 70% ethanol particle processed, and dry, compressing tablet, makes 500, every heavy 0.35g.
Embodiment 3
Get wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g,Red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g, above ten tastes, get Radix Angelicae Sinensis 120g, and red sage root 120g joinsCO2In supercritical extract device, ethanol is as entrainer, and the percent by volume that entrainer accounts for total extractant is 5%, extractionPressure power 20MPa, 40 DEG C of temperature, CO2Flow 2ml/g crude drug min, extraction time 160min, obtains overcritical extractionGet thing, add starch, 70% ethanol particle processed, dry, compressing tablet, makes 500, every heavy 0.35g.
Embodiment 4: active pain relieving tablet suppresses the experimental study data of C6 cell proliferation
1 experiment material
1.1 experiment cell lines
Rat neurogliocytoma cells (C6), Nanjing Medical University's laboratory cell bank, DMEM+10%FBS routineCultivate.
1.2 Experimental agents
Drugs: the active pain relieving tablet of the present invention: press embodiment 3 method preparations.
Liquid liquid storage: take the active pain relieving tablet of 100mg, be dissolved in 5ml absolute ethyl alcohol, 0.2 μ m filter filters, 500 μ lThe packing of doff pipe ,-20 DEG C of storages, 0.2 μ m filter filters the use of absolute ethyl alcohol in order to control group simultaneously.
1.3 experiment reagent
The Cat.No.12100-061Lot.No.758137 of DMEM(GIBCO company); Hyclone (Hangzhoupro, sky, ZhejiangThe Lot.No.100419 of bio tech ltd); NaHCO3(Shanghai hundred million Cat. of chemical reagent Co., Ltd of a specified durationNo.11810-033Lot.No.1088387); Trypsin(AMRESCO company lot number: 2010/04); EDTA(AMRESCO company lot number: 2009/10); PenicillinGSodiumSalt(AMRESCO company lot number:2010242); StreptomycinSulfate(AMRESCO company lot number: 2010382); Absolute ethyl alcohol (NanjingLearn reagent Co., Ltd 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 (U.S. MDCompany's model: SPECTRAMAX190); CO2 incubator (FORMA model: 3111); Super-clean bench (Su JingSafe and sound company of group manufactures model: 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 Co., Ltd model:BT323S); Full-automatic high-pressure autoclave (Japanese SANYO company model: MLS-3020); Table electrothermal air blast is dryDry case (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, pipette, Tips are some.
2 experimental techniques
1) C6 cell carries out cellar culture (10cm culture dish) with DMEM+10%FBS in 37 DEG C, 5%CO2, whenGrowth of Cells is during to logarithmic phase, and collecting cell, discards nutrient solution, and PBS fine laundering 3 times, adds 3ml0.25% tryptoseEnzyme-0.04%EDTA, after 37 DEG C of digestion 2min, adds 5ml complete medium neutralization reaction wherein, and piping and druming is thinAfter born of the same parents, proceeded in centrifuge tube, the centrifugal 5min of 1000rpm, adjusts 3 × 104/ml of concentration of cell suspension.
2) cell kind is entered in 96 well culture plates, every hole adds cell suspension 180 μ l, and culture plate is put into cell culture incubator(37 DEG C, 5%CO2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add active pain relieving tablet solution, continue to cultivate 24h.
4) after 24h, add 20 μ lMTT solution (5mg/ml, i.e. 0.5%MTT), continue to cultivate 4h.
5) after 4h, buckle method is removed supernatant, pats dry gently with blotting paper, and every hole adds 200 μ l dimethyl sulfoxide (DMSO)s, puts on shaking tableLow-speed oscillation 10min, fully dissolves crystal. Measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument 490nm place.
6) background (do not add cell, only add nutrient solution) is set simultaneously, control wells (the medicine dissolving medium of cell, same concentrations,Nutrient solution, MTT, dimethyl sulfoxide (DMSO)), set 6 multiple holes for every group.
7) result represents the inhibiting rate of cell with medicine:
Cell increment inhibiting rate (%)=(control wells OD value-dosing holes OD value)/control wells OD value × 100%. Experiment is heavyMultiple 3 times.
3 statistical dispositions
Adopt correlation analysis and Studentt inspection in MicrosoftExcel2003 software, data are with mean ± S.D.Represent.
4 experimental results
Statistical result showed after mtt assay experiment, with control group comparison, in the time that dosage reaches 5mg/ml, to C6 cellPropagation suppresses variant (P < 0.05), and dosage this difference in the time of 10mg/ml has conspicuousness (P < 0.01), when dosage reachesDuring to 15-20mg/ml, there is utmost point significant difference (P < 0.001).
The active pain relieving tablet of table 1 is on C6 cell inhibitory effect impact research (X ± SD)
Note: with control group comparison, * P < 0.01; * P < 0.001
5 experiment conclusion
Active pain relieving tablet can suppress C6 cell proliferation, reduces the Growth of Cells number of C6 cell, and this effect is dosageDependence.
Claims (3)
1. active pain relieving tablet suppresses the application in rat neurogliocytoma cells C6 cell proliferation medicine, its feature in preparationBe that active pain relieving tablet is by wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, ring perfume (or spice)Attached 20g, red sage root 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g make as bulk drug, and preparation method is by the following step groupBecome: get wilsonii 100g, root of Chinese clematis 100g, Radix Angelicae Sinensis 150g, aconiti preparata,radix 40g, wild aconite root 40g, Radde Anemone Rhizome 20g, pelletGinseng 150g, frankincense 15g, myrrh 15g, Chinese ephedra 30g join CO2In supercritical extract device, ethanol, as entrainer, is carried secretlyThe percent by volume that agent accounts for total extractant is 4-6%, extracting pressure 15-30MPa, temperature 30-50 DEG C, CO2Flow 1-3m1/gCrude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, and 70% ethanol particle processed is dry, pressesSheet, makes 500, every heavy 0.35g.
2. a kind of active pain relieving tablet suppresses rat neurogliocytoma cells C6 cell proliferation medicine in preparation according to claim 1Application in thing, is characterized in that CO described in the preparation method of active pain relieving tablet2Supercritical extract entrainer accounts for total extractantPercent by volume be 5%.
3. a kind of active pain relieving tablet suppresses rat neurogliocytoma cells C6 cell proliferation medicine in preparation according to claim 1Application in thing, is characterized in that CO described in the preparation method of active pain relieving tablet2The extracting pressure 20MPa of supercritical extract,40 DEG C of temperature, CO2Flow 2ml/g crude drug min, extraction time 160min.
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| CN106214770B (en) * | 2016-09-18 | 2020-04-10 | 天地恒一制药股份有限公司 | Application of collateral activating and pain relieving capsule in preparation of medicine for treating diabetic peripheral neuropathy |
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Inventor after: Jiang Jinyan Inventor before: Gao Zhongqing |
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Effective date of registration: 20160419 Address after: 266035 Shandong province Qingdao City, Hefei Road No. 758 Applicant after: QILU HOSPITAL OF SHANDONG UNIVERSITY (QINGDAO) Address before: Industrial Park No. 87 Yu Bridge 255086 Shandong province Zibo high tech Zone Wanjie Road Room 301 Applicant before: Zibo Qidingli Patent Information Consulting Co., Ltd. |
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Effective date of registration: 20200427 Address after: 363000 No.16, Weihui Road, Suian Industrial Zone, Zhangpu County, Zhangzhou City, Fujian Province Patentee after: Fujian tongyitangguo Pharmaceutical Development Co., Ltd Address before: 266035 Shandong province Qingdao City, Hefei Road No. 758 Patentee before: QILU HOSPITAL OF SHANDONG UNIVERSITY (QINGDAO) |