CN103610794B - Preparation method and application of nerve-calming tablets - Google Patents
Preparation method and application of nerve-calming tablets Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 17
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 13
- 230000004663 cell proliferation Effects 0.000 claims abstract description 11
- 201000009410 rhabdomyosarcoma Diseases 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 229940079593 drug Drugs 0.000 claims description 13
- 241001619461 Poria <basidiomycete fungus> Species 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 9
- 239000008187 granular material Substances 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 6
- 235000019698 starch Nutrition 0.000 claims description 6
- 239000008107 starch Substances 0.000 claims description 6
- 230000001629 suppression Effects 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 abstract description 3
- 241000382455 Angelica sinensis Species 0.000 abstract 1
- 241000092665 Atractylodes macrocephala Species 0.000 abstract 1
- 244000303040 Glycyrrhiza glabra Species 0.000 abstract 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 abstract 1
- 241000112528 Ligusticum striatum Species 0.000 abstract 1
- 244000292697 Polygonum aviculare Species 0.000 abstract 1
- 235000006386 Polygonum aviculare Nutrition 0.000 abstract 1
- 244000197580 Poria cocos Species 0.000 abstract 1
- 235000008599 Poria cocos Nutrition 0.000 abstract 1
- 241000607122 Uncaria tomentosa Species 0.000 abstract 1
- 239000004480 active ingredient Substances 0.000 abstract 1
- 235000011472 cat’s claw Nutrition 0.000 abstract 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 abstract 1
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- 238000002474 experimental method Methods 0.000 description 8
- 238000000034 method Methods 0.000 description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
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- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- 241000345998 Calamus manan Species 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
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- FCPVYOBCFFNJFS-LQDWTQKMSA-M benzylpenicillin sodium Chemical compound [Na+].N([C@H]1[C@H]2SC([C@@H](N2C1=O)C([O-])=O)(C)C)C(=O)CC1=CC=CC=C1 FCPVYOBCFFNJFS-LQDWTQKMSA-M 0.000 description 1
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a preparation method of nerve-calming tablets. The nerve-calming tablets are prepared from active ingredients: 90g of radix bupleuri, 135g of angelica sinensis, 90g of ligusticum wallichii, 180g of poria cocos, 180g of uncaria tomentosa, 180g of vine of multiflower knotweed, 180g of fried atractylodes macrocephala and 68g of liquorice, and is prepared in a supercritical extraction mode. Therefore, the content is greatly increased, and the dosage is reduced. The invention further provides an application of the nerve-calming tablets in preparing a medicament for inhibiting rhabdomyosarcoma A-204 cell 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 ANLE PIAN.
Background technology
ANLE PIAN is recorded in Ministry of Public Health standard WS3-B-0274-90, prescription is Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g, above eight tastes, hook taking rattan, Radix Bupleuri half and half amount, be ground into fine powder, sieve; The Six-elements such as second half and all the other Poria decoct with water secondary, each 2 hours, collecting decoction, and filter, filtrate is concentrated into the clear paste that relative density is about 1.28, add the mixing of above-mentioned powder, make granule, dry, are pressed into 1000, sugar coating, obtain final product.Energy soothing liver-QI for relieving depression, arresting convulsion is calmed the nerves.For spirit depressing, have a sleepless night in terror, discomfort uncomfortable in chest, anorexia Mental fatigue, also can use neurosis, climacteric syndrome and the nocturnal fretfulness in infants, the symptom such as to grind one's teeth in sleep.
In prior art, not yet there is ANLE PIAN extracting the report adopting supercritical technology in preparation, and adopt the method for beating powder and soak by water, 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.
In prior art, the every sheet 0.5g of ANLE PIAN, each 4-6 sheet, 3 times on the one, adopt the every sheet 0.5g of ANLE PIAN that the present invention is prepared into, but the medical material amount contained is original 2 times, therefore only need 2-3 sheet at every turn, within 1st, take 3 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 ANLE PIAN.
Another object of the present invention is to provide a kind of ANLE PIAN to suppress the application in rhabdomyosarcoma A-204 cell proliferation in preparation.
Technical scheme: the object of the invention is by following scheme realize:
A kind of preparation method of ANLE PIAN, be made up as crude drug of Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g, described method is made up of the following step: get Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g, 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 1-3ml/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
The preparation method of above-mentioned a kind of ANLE 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 ANLE 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 ANLE PIAN suppresses the application in rhabdomyosarcoma A-204 cell proliferation in preparation, ANLE PIAN is made up as crude drug of Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g, preparation method is made up of the following step: get Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g, 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 1-3ml/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Above-mentioned ANLE PIAN suppresses the application in rhabdomyosarcoma A-204 cell proliferation in preparation, CO described in ANLE PIAN preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned ANLE PIAN suppresses the application in rhabdomyosarcoma A-204 cell proliferation in preparation, it is characterized in that CO described in ANLE PIAN preparation method
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 Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g join in CO2 supercritical extraction device, ethanol is as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, extracting pressure 15MPa, temperature 30 DEG C, CO
2flow 1ml/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 Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g 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 3ml/g crude drug min, extraction time 180min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 3
Get Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g 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 2ml/g crude drug min, extraction time 160min, obtains supercritical extract, adds starch, 70% ethanol granule, and dry, tabletting, makes 500, the heavy 0.5g of every sheet.
Embodiment 4: ANLE PIAN suppresses the experimentation data of A-204 cell proliferation
1 experiment material
1.1 experiment cell strains
Rhabdomyosarcoma (A-204), Nanjing Medical University's laboratory cell storehouse, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: ANLE PIAN of the present invention: prepare by embodiment 3 method.
Medicinal liquid liquid storage: take 100mg ANLE PIAN, is dissolved in 5ml dehydrated alcohol, 0.2 μm of frit, 500 μ l doff 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) A-204 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 ANLE 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 A-204 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 ANLE PIAN is to A-204 cell inhibitory effect influence research
Note: compare with matched group, * P<0.01; * P<0.001
5 experiment conclusion
ANLE PIAN can suppress A-204 cell proliferation, and reduce the Growth of Cells number of A-204 cell, this effect is dose dependent.
Claims (3)
1. the application of ANLE PIAN in preparation suppression rhabdomyosarcoma A-204 cell proliferation, it is characterized in that ANLE PIAN is by Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g makes as crude drug, preparation method is made up of the following step: get Radix Bupleuri 90g, Radix Angelicae Sinensis 135g, Rhizoma Chuanxiong 90g, Poria 180g, Ramulus Uncariae Cum Uncis 180g, Caulis Polygoni Multiflori 180g, Rhizoma Atractylodis Macrocephalae (parched) 180g, Radix Glycyrrhizae 68g, 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, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add starch, 70% ethanol granule, dry, tabletting, make 500, the heavy 0.5g of every sheet.
2. a kind of ANLE PIAN suppresses the application in rhabdomyosarcoma A-204 cell proliferation in preparation according to claim 1, it is characterized in that CO described in ANLE PIAN preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of ANLE PIAN suppresses the application in rhabdomyosarcoma A-204 cell proliferation in preparation according to claim 1, it is characterized in that CO described in ANLE PIAN preparation method
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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| CN102824438A (en) * | 2012-09-20 | 2012-12-19 | 南京正亮医药科技有限公司 | Preparation method of Anle tablets |
| CN102988502A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of Desheng tablet |
| CN102988522A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of naodesheng tablets |
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| CN102824438A (en) * | 2012-09-20 | 2012-12-19 | 南京正亮医药科技有限公司 | Preparation method of Anle tablets |
| CN102988502A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of Desheng tablet |
| CN102988522A (en) * | 2012-10-15 | 2013-03-27 | 李正梅 | Preparation method and application of naodesheng tablets |
Non-Patent Citations (3)
| Title |
|---|
| 中药治疗恶性肿瘤的多种治法体现;张钢亭等;《中国药师》;20121231;第15卷(第8期);第1194-1196页，尤其第1195页右栏第2段 * |
| 周金黄等.白术的抗肿瘤作用及其免疫学机理.《中药药理与临床研究进展 第一册》.中国科学技术出版社,1992,(第1版),第329-330页. * |
| 郁仁存.横纹肌肉瘤.《郁仁存中西医结合肿瘤学》.中国协和医科大学出版社,2008,(第1版),第422页. * |
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