CN103919885A - Preparation method and application of nasosinusitis soft capsule - Google Patents
Preparation method and application of nasosinusitis soft capsule Download PDFInfo
- Publication number
- CN103919885A CN103919885A CN201410169286.6A CN201410169286A CN103919885A CN 103919885 A CN103919885 A CN 103919885A CN 201410169286 A CN201410169286 A CN 201410169286A CN 103919885 A CN103919885 A CN 103919885A
- Authority
- CN
- China
- Prior art keywords
- nasosinusitis
- soft capsule
- preparation
- flos
- crude drug
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000007901 soft capsule Substances 0.000 title claims abstract description 75
- 238000002360 preparation method Methods 0.000 title claims abstract description 44
- 241000628997 Flos Species 0.000 claims abstract description 61
- 239000003814 drug Substances 0.000 claims abstract description 57
- 229940079593 drug Drugs 0.000 claims abstract description 37
- 238000000194 supercritical-fluid extraction Methods 0.000 claims abstract description 24
- 239000009670 cang er zi wan Substances 0.000 claims abstract description 23
- 235000012424 soybean oil Nutrition 0.000 claims abstract description 20
- 239000003549 soybean oil Substances 0.000 claims abstract description 20
- 230000004663 cell proliferation Effects 0.000 claims abstract description 14
- 208000032612 Glial tumor Diseases 0.000 claims abstract description 13
- 206010018338 Glioma Diseases 0.000 claims abstract description 13
- 206010010774 Constipation Diseases 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 35
- 238000000605 extraction Methods 0.000 claims description 18
- 239000000284 extract Substances 0.000 claims description 13
- 239000000084 colloidal system Substances 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 241000205585 Aquilegia canadensis Species 0.000 abstract 1
- 240000005250 Chrysanthemum indicum Species 0.000 abstract 1
- 235000018959 Chrysanthemum indicum Nutrition 0.000 abstract 1
- 241000218378 Magnolia Species 0.000 abstract 1
- 241000123069 Ocyurus chrysurus Species 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 229960004756 ethanol Drugs 0.000 description 12
- 241000699670 Mus sp. Species 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 9
- RBRPTFMVULVGIC-ZTIIIDENSA-N Xanthatin Chemical compound C1C=C(\C=C\C(C)=O)[C@@H](C)C[C@@H]2OC(=O)C(=C)[C@H]21 RBRPTFMVULVGIC-ZTIIIDENSA-N 0.000 description 7
- 230000000968 intestinal effect Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- RBRPTFMVULVGIC-UHFFFAOYSA-N xanthatin Natural products C1C=C(C=CC(C)=O)C(C)CC2OC(=O)C(=C)C21 RBRPTFMVULVGIC-UHFFFAOYSA-N 0.000 description 7
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000007788 liquid Substances 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
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 239000003610 charcoal Substances 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 239000013558 reference substance Substances 0.000 description 3
- 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
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 238000006073 displacement reaction Methods 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000003304 gavage Methods 0.000 description 2
- 238000005325 percolation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000006424 Flood reaction Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 229920002334 Spandex Polymers 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000001785 acacia senegal l. willd gum Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- 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
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012738 dissolution medium Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000005176 gastrointestinal motility Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000004900 laundering Methods 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000021590 normal diet Nutrition 0.000 description 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000004759 spandex Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229960002385 streptomycin sulfate Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012353 t test Methods 0.000 description 1
- 235000019640 taste Nutrition 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a preparation method of a nasosinusitis soft capsule. The nasosinusitis soft capsule is prepared by taking 1344g of fructus xanthii, 252g of flos magnolia, 84g of honeysuckle, 84g of madder, 84g of chrysanthemum indicum and proper amount of soybean oil as raw material medicines through supercritical extraction, so that content is greatly improved and dose is reduced. The invention further provides application of the nasosinusitis soft capsule in preparing drugs for inhibiting cell proliferation of human glioma cells SF295 and drugs for inhibiting constipation.
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 soft capsule for nasosinusitis.
Background technology
Soft capsule for nasosinusitis is recorded in ministry standard, prescription for Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil appropriate, the above five tastes, Flos Lonicerae, Flos Chrysanthemi Indici, Flos Magnoliae extract volatile oil 4 hours, the another device of aqueous solution after distillation is collected, medicinal residues and Fructus Xanthii add 8 times of water gagings and decoct secondary, each 3 hours, collecting decoction, filtered, filtrate and above-mentioned aqueous solution merge, being concentrated into relative density is 1.10～1.20 (80 ℃), adds ethanol and makes to be 60%, to stir containing alcohol amount, standing, filter; Precipitation is used 60% washing with alcohol, and washing liquid is incorporated in filtrate.Radix Rubiae, according to the percolation (appendix I O of Chinese Pharmacopoeia version in 2010) under fluid extract and extractum item, makes solvent with 70% ethanol, floods after 24 hours, slowly percolation, collects 12 times of amount percolates, merges with above-mentioned filtrate, reclaim ethanol and be concentrated into the thick paste that relative density is about 1.35 (60 ℃), spraying into above-mentioned volatile oil, adding soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
In prior art, not yet there is soft capsule for nasosinusitis aspect extraction preparation, adopting the report of supercritical technology, and the method that adopts vapor distillation and decocting to boil, 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 soft capsule for nasosinusitis.
Another object of the present invention is to provide a kind of soft capsule for nasosinusitis to suppress the application in human glioma cell's SF295 cell proliferation medicine and constipation medicine in preparation.
Technical scheme: the object of the invention is to realize by following scheme:
A kind of preparation method of soft capsule for nasosinusitis, by Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil, made in right amount as crude drug, described method is comprised of the following step: Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
The preparation method of above-mentioned a kind of soft capsule for nasosinusitis, 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 soft capsule for nasosinusitis, 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 soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine in preparation, soft capsule for nasosinusitis is by by Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil is made as crude drug in right amount, preparation method is comprised of the following step: get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
Above-mentioned a kind of soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine, CO described in soft capsule for nasosinusitis preparation method in preparation
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned a kind of soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine, CO described in soft capsule for nasosinusitis preparation method in preparation
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
The application of above-mentioned soft capsule for nasosinusitis in preparation treatment constipation medicine, soft capsule for nasosinusitis is by by Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil is made as crude drug in right amount, preparation method is comprised of the following step: get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
Above-mentioned soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine, CO described in soft capsule for nasosinusitis preparation method in preparation
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
Above-mentioned soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine, CO described in soft capsule for nasosinusitis 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 0.34g of former soft capsule for nasosinusitis, each 3-4 grain, 3 times on the one, the every 0.3g of soft capsule for nasosinusitis that adopts the present invention to be prepared into, but the medical material amount containing is approximately original 2 times, therefore only needs 2 at every turn, within 1st, take 3 times, greatly reduced dose having under the condition of more active component.
The comparison of Xanthatin content in soft capsule for nasosinusitis prepared by test one, distinct methods
1, instrument and reagent soft capsule for nasosinusitis of the present invention: press embodiment 3 methods preparations, get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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, 40 ℃ of temperature, CO
2flow 2m1/g crude drug? min, extraction time 160min, obtains supercritical extract, adds soybean oil appropriate, mixes, and with colloid mill, grinds, and is chilled to room temperature, is pressed into 1000 of soft capsules, dry, deoils, and dries, and obtains every 0.3g.Former soft capsule for nasosinusitis, by the preparation of ministry standard method, gets Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, through extracting, makes 1000, dry, deoils, and dries, and obtains every 0.34g.Agilent1200 high performance liquid chromatograph; METTLER AE240 electronic analytical balance; Xanthatin standard substance (Nat'l Pharmaceutical & Biological Products Control Institute).
2, method
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica, be filler; Methanol-water (55:45) is mobile phase; Detection wavelength is 278nm.Number of theoretical plate calculates by Xanthatin peak, should be not less than 3000.
The preparation of reference substance solution: precision takes at 60 ℃ of drying under reduced pressure Xanthatin reference substance of 4 hours appropriate, adds methanol and makes every 1ml containing the solution of 18 μ g, obtains.
The preparation of need testing solution: get soft capsule for nasosinusitis of the present invention and former soft capsule for nasosinusitis, porphyrize, mixes, and gets 1g, accurately weighed, precision adds 70% ethanol 20ml, close plug, supersound process 10 minutes, centrifugal, get supernatant, obtain.
Algoscopy is accurate reference substance solution and each 20 μ l of need testing solution of drawing respectively, and injection liquid chromatography, measures, and obtains.
3, result
Result shows, in soft capsule for nasosinusitis of the present invention, the content of Xanthatin is 0.89mg/ grain; And the content of Xanthatin is 0.43mg/ grain in former soft capsule for nasosinusitis, in the situation that dose reduces, Xanthatin content improves a lot.
Above-mentioned research shows, the soft capsule for nasosinusitis that adopts the present invention to prepare, and active constituent content is higher than the standby soft capsule for nasosinusitis of ministry standard legal system, and dose can reduce.
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 realizing based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1
Get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins in CO2 supercritical extraction device, ethanol is as entrainer, 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, obtains supercritical extract, adds soybean oil appropriate, mixes, and with colloid mill, grinds, and is chilled to room temperature, is pressed into 1000 of soft capsules, dry, deoils, and dries, and obtains.
Embodiment 2
Get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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 soybean oil appropriate, mixes, and with colloid mill, grinds, and is chilled to room temperature, is pressed into 1000 of soft capsules, dry, deoils, and dries, and obtains.
Embodiment 3
Get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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, 40 ℃ of temperature, CO
2flow 2m1/g crude drug min, extraction time 160min, obtains supercritical extract, adds soybean oil appropriate, mixes, and with colloid mill, grinds, and is chilled to room temperature, is pressed into 1000 of soft capsules, dry, deoils, and dries, and obtains.
Test two: soft capsule for nasosinusitis suppresses the experimentation data of SF295 cell proliferation
1 experiment material
1.1 experiment cell strains
Human glioma cell SF295 cell, Nanjing Medical University's laboratory cell bank, DMEM+10%FBS cellar culture.
1.2 Experimental agents
Drugs: soft capsule for nasosinusitis of the present invention: press embodiment 3 method preparations.
Medicinal liquid liquid storage: take 100mg soft capsule for nasosinusitis, be dissolved in 5ml dehydrated alcohol, 0.2 μ m filter filters, 500 μ ldoff pipe subpackages ,-20 ℃ of storages, 0.2 μ m filter filters dehydrated alcohol in order to the use of matched group simultaneously.The control sample of preparing Fructus Xanthii by same method.
1.3 experiment reagent
DMEM (Cat.No.12100-061Lot.No.758137 of GIBCO company); Hyclone (Lot.No.100419 of Tian Hang bio tech ltd, Zhejiang); NaHCO3 (the Cat. No.11810-033Lot.No.1088387 of Shanghai Jiu Yi chemical reagent company limited); 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 (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) SF295 cell carries out cellar culture (10cm culture dish) with DMEM+10%FBS in 37 ℃, 5%CO2, when Growth of Cells is during to logarithmic (log) phase, collecting cell, discards 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 piping and druming cell, 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, culture plate put into cell culture incubator (37 ℃, 5%CO2) cellar culture.
3) according to Growth of Cells situation, generally grow to 50%-70%, add the control sample of soft capsule for nasosinusitis solution or Fructus Xanthii, continue to cultivate 24h.
4) after 24h, add 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT), continue to cultivate 4h.
5) after 4h, 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.At enzyme-linked immunosorbent assay instrument 490nm place, measure the light absorption value in each hole.
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), sets 6 multiple holes for every group.
7) result represents the suppression ratio of cell with medicine:
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 represent with mean ± S.D..
4 experimental results
Statistical result showed after mtt assay experiment, with matched group comparison, when dosage reaches 5mg/ml, to SF295 cell inhibitory effect variant (P<0.05), dosage this difference when 10mg/ml has significance (P<0.01), has utmost point significant difference (P<0.001) when dosage reaches 15-20mg/ml.
Table 1 soft capsule for nasosinusitis is on SF295 cell inhibitory effect impact research (X ± SD)
Note: with matched group comparison, * P<0.01; * P<0.001, with the comparison of Herba Xanthii subgroup,
#p<0.01;
##p<0.001
5 experiment conclusion
Soft capsule for nasosinusitis can suppress SF295 cell proliferation, reduces the Growth of Cells number of SF295 cell, and this effect is dose dependent, and compares and also have significant difference with single Herba Xanthii subgroup.
Test three: the impact of soft capsule for nasosinusitis of the present invention on the motion of mice with constipation intestinal propulsion
1, animal: healthy ICR mice, male and female half and half, heavy 18-22g, is provided production licence number SCXK (Soviet Union) 2002-0031 by Nanjing Medical University's animal center.
2, medicine and reagent: press above-described embodiment 3 preparation soft capsule for nasosinusitis of the present invention, get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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, 40 ℃ of temperature, CO
2flow 2m1/g crude drug min, extraction time 160min, obtain supercritical extract, add soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain 1000 soft capsules, 0.3 gram every, in every soft capsule, contain crude drug 1.8g, by Yunnan Yunlong Pharmaceutical Co., Ltd., provided, lot number 20140611, clinical people's consumption per day is each 10, be that clinical people's consumption per day is 18g crude drug, therefore determine that every mice consumption is 18x0.0026=0.05g crude drug, with a kilogram calculating, dosage is 2.5g crude drug/kg, this is low dosage, dosage 5g crude drug/kg is middle dosage, dosage 10g crude drug/kg is high dose.Phenolphthalein tablets (phenolphthalein sugar-tablet), Junan, Shandong Province pharmaceutical factory, specification: 100mg*100 sheet/box, lot number 20140506, people's consumption per day 0.8g/ days, being equivalent to every mice consumption is 0.8gx0.0026=0.002g, with a kilogram calculating, dosage is 0.1g/kg, and the 3 times of amounts of take are dosage, and positive group gives phenolphthalein tablets 0.3g/kg.All the other reagent are domestic analytical pure.R-1132, Wujin pharmaceutical factory, 20130724.
3, process of the test: get 60 of ICR mices, with the modeling of R-1132 50m/kg gavage, continuous 10 days, normal diet during this time, form Constipation Model, random packet, every group approximately 10, male and female half and half, be divided into normal group, model group, positive drug group, soft capsule of the present invention is high, in, low three dosage groups, administration is the same, successive administration 14d, before experiment, water 12h is can't help in fasting, 2h after last administration, the charcoal that is 5% by volume fraction respectively end arabic gum suspension lml/g gavages, after 20min, put to death and respectively organize mice, cut open the belly and measure every Mus small intestinal charcoal end displacement and small intestinal total length, be converted into the percentage rate that charcoal end displacement accounts for small intestinal total length, t test and judge group difference, the results are shown in Table 2.
The impact (n=10) of table 2 soft capsule of the present invention on the motion of mice with constipation intestinal propulsion
With model group comparison, * * P<0.01*P<0.05, with the comparison of positive drug group, ##P<0.01#P<0.05
4, result: medicine group of the present invention and model group comparison, there is obvious promotion mice with constipation intestinal propulsion motion, promote gastrointestinal motility in mice, there is significant difference, with positive drug comparison, medicine of the present invention senior middle school dosage group is promoting to have significant difference aspect the motion of mice with constipation intestinal propulsion, and effect is better than positive drug.
Claims (9)
1. the preparation method of a soft capsule for nasosinusitis, by Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil, as crude drug, made in right amount, it is characterized in that described method is comprised of the following step: Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtains supercritical extract, adds soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
2. a kind of preparation method of soft capsule for nasosinusitis according to claim 1, is characterized in that described CO
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
3. a kind of preparation method of soft capsule for nasosinusitis 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 soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine in preparation according to claim 1, it is characterized in that soft capsule for nasosinusitis is by by Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil is made as crude drug in right amount, preparation method is comprised of the following step: get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
5. a kind of soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in soft capsule for nasosinusitis preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
6. a kind of soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in soft capsule for nasosinusitis preparation method
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
7. the application of soft capsule for nasosinusitis according to claim 1 in preparation treatment constipation medicine, it is characterized in that soft capsule for nasosinusitis is by by Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g, soybean oil is made as crude drug in right amount, preparation method is comprised of the following step: get Fructus Xanthii 1344g, Flos Magnoliae 252g, Flos Lonicerae 84g, Radix Rubiae 84g, Flos Chrysanthemi Indici 84g joins 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 ℃, CO2 flow 1-3m1/g crude drug min, extraction time 150-180min, obtain supercritical extract, add soybean oil appropriate, mix, with colloid mill, grind, be chilled to room temperature, be pressed into 1000 of soft capsules, dry, deoil, dry, obtain.
8. a kind of soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in soft capsule for nasosinusitis preparation method
2the percent by volume that supercritical extraction entrainer accounts for total extractant is 5%.
9. a kind of soft capsule for nasosinusitis suppresses the application in human glioma cell SF295 cell proliferation medicine in preparation according to claim 4, it is characterized in that CO described in soft capsule for nasosinusitis preparation method
2the extracting pressure 20MPa of supercritical extraction, 40 ℃ of temperature, CO
2flow 2ml/g crude drug min, extraction time 160min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410169286.6A CN103919885B (en) | 2014-04-25 | 2014-04-25 | A kind of preparation method of soft capsule for nasosinusitis and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410169286.6A CN103919885B (en) | 2014-04-25 | 2014-04-25 | A kind of preparation method of soft capsule for nasosinusitis and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103919885A true CN103919885A (en) | 2014-07-16 |
| CN103919885B CN103919885B (en) | 2015-09-30 |
Family
ID=51138427
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410169286.6A Active CN103919885B (en) | 2014-04-25 | 2014-04-25 | A kind of preparation method of soft capsule for nasosinusitis and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103919885B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104840797A (en) * | 2015-05-28 | 2015-08-19 | 吉林省正和药业集团股份有限公司 | Preparation method and application of Yixuanning capsule |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1634263A (en) * | 2004-11-03 | 2005-07-06 | 贵州海泰药业技术有限公司 | Soft capsule for nasosinusitis and its preparation |
| CN101569677A (en) * | 2009-03-16 | 2009-11-04 | 云南云龙制药有限公司 | Chinese patent medicament for treating conjunctival congestion and preparation method thereof |
| KR20130116500A (en) * | 2012-04-16 | 2013-10-24 | 김현정 | Korean medical compositions for improving rhinitis and empyema and the set |
| CN103494885A (en) * | 2013-10-11 | 2014-01-08 | 南京正亮医药科技有限公司 | Preparation method and application of orifice-freeing rhinitis tablets |
-
2014
- 2014-04-25 CN CN201410169286.6A patent/CN103919885B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1634263A (en) * | 2004-11-03 | 2005-07-06 | 贵州海泰药业技术有限公司 | Soft capsule for nasosinusitis and its preparation |
| CN101569677A (en) * | 2009-03-16 | 2009-11-04 | 云南云龙制药有限公司 | Chinese patent medicament for treating conjunctival congestion and preparation method thereof |
| KR20130116500A (en) * | 2012-04-16 | 2013-10-24 | 김현정 | Korean medical compositions for improving rhinitis and empyema and the set |
| CN103494885A (en) * | 2013-10-11 | 2014-01-08 | 南京正亮医药科技有限公司 | Preparation method and application of orifice-freeing rhinitis tablets |
Non-Patent Citations (2)
| Title |
|---|
| 俞发荣等: "苍耳子药物血清对H4细胞毒性作用的实验研究", 《中国临床研究》 * |
| 金雪梅等: "天然蒽醌抗肿瘤作用机制研究进展", 《延边大学医学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104840797A (en) * | 2015-05-28 | 2015-08-19 | 吉林省正和药业集团股份有限公司 | Preparation method and application of Yixuanning capsule |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103919885B (en) | 2015-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103494878A (en) | Preparation method and application of compound danshen tablets | |
| CN102988526B (en) | Method for preparing Bailing tablets and application | |
| CN102988612B (en) | Preparation method and application of tablet for clearing away heat | |
| CN102988480B (en) | Method for preparing bile and patchouli rhinitis tablets and application | |
| CN102988665B (en) | Method for preparing gynecological menstruation regulating tablets and application | |
| CN102988545B (en) | Preparation method and application of Yulian tablet | |
| CN102988489B (en) | Preparation method and application of nose comforting tablet | |
| CN103028005B (en) | Preparation method of anti-lumbago tablet and application thereof | |
| CN103495117A (en) | Preparation method and application of thrombus removing and collaterals dredging tablets | |
| CN102988614B (en) | Preparation method and application of antiphlogosis tablet | |
| CN102988479B (en) | Preparation method and application of anti-dazzling tablet | |
| CN102988550B (en) | Preparation method and application of qinlian tablet | |
| CN103977391A (en) | Preparation method and applications of bupleurum tenue capsule | |
| CN103919885B (en) | A kind of preparation method of soft capsule for nasosinusitis and application | |
| CN103494885A (en) | Preparation method and application of orifice-freeing rhinitis tablets | |
| CN102836381A (en) | Preparation method and application of middle calming tablet | |
| CN102988611B (en) | Preparation method and application of antihypertension tablet | |
| CN102988500B (en) | Method for preparing compound Dantong tablets and application | |
| CN102988881B (en) | Preparation method and application of pulse unblocking tablet | |
| CN103479749A (en) | Preparation method and application of Gegenqinlian tablet | |
| CN102988522B (en) | Preparation method and application of naodesheng tablets | |
| CN103735960A (en) | Application of boenninghausenia sessilicarpa cough-asthma tablet to prepare medicaments for inhibiting lung cancer cell LLC proliferation | |
| CN102988691B (en) | Preparation method and application of liver-strengthening tablet | |
| CN102988748B (en) | Preparation method and application of yin tonifying and blood sugar reducing tablet | |
| CN103705825A (en) | Application of lespedeza cuneata turbidity removing tablet in preparation of medicine for inhibiting cell proliferation of embryonal carcinoma F9 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CP02 | Change in the address of a patent holder |
Address after: No. 5 Jinghong Road, Changhong Community, Ala Street Office, Kunming Economic Development Zone, China (Yunnan) Pilot Free Trade Zone, Kunming City, Yunnan Province, 650217 Patentee after: YUNNAN YUNLONG PHARMACEUTICAL Co.,Ltd. Address before: No.5 Jinghong Road, Kunming Economic and Technological Development Zone, Yunnan Province Patentee before: YUNNAN YUNLONG PHARMACEUTICAL Co.,Ltd. |
|
| CP02 | Change in the address of a patent holder |