CN101716133A - Aseculin ophthalmic gel and production method and application thereof - Google Patents
Aseculin ophthalmic gel and production method and application thereof Download PDFInfo
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- CN101716133A CN101716133A CN200910242109A CN200910242109A CN101716133A CN 101716133 A CN101716133 A CN 101716133A CN 200910242109 A CN200910242109 A CN 200910242109A CN 200910242109 A CN200910242109 A CN 200910242109A CN 101716133 A CN101716133 A CN 101716133A
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- 229940100655 ophthalmic gel Drugs 0.000 title claims abstract description 32
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 238000002360 preparation method Methods 0.000 claims abstract description 14
- 238000000605 extraction Methods 0.000 claims abstract description 11
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000004327 boric acid Substances 0.000 claims abstract description 10
- LDCRTTXIJACKKU-ONEGZZNKSA-N dimethyl fumarate Chemical compound COC(=O)\C=C\C(=O)OC LDCRTTXIJACKKU-ONEGZZNKSA-N 0.000 claims abstract description 10
- 239000000203 mixture Substances 0.000 claims abstract description 9
- 229960004419 dimethyl fumarate Drugs 0.000 claims abstract description 8
- 239000000284 extract Substances 0.000 claims abstract description 7
- -1 hydroxypropyl Chemical group 0.000 claims description 25
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 239000008589 Cortex Fraxini Substances 0.000 claims description 16
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 239000013078 crystal Substances 0.000 claims description 15
- 239000000758 substrate Substances 0.000 claims description 14
- 239000000706 filtrate Substances 0.000 claims description 12
- 235000010413 sodium alginate Nutrition 0.000 claims description 11
- 239000000287 crude extract Substances 0.000 claims description 10
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 9
- 229940005550 sodium alginate Drugs 0.000 claims description 9
- 239000000661 sodium alginate Substances 0.000 claims description 9
- 238000004090 dissolution Methods 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 206010015943 Eye inflammation Diseases 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 6
- 230000008020 evaporation Effects 0.000 claims description 6
- 238000004440 column chromatography Methods 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 4
- 239000012982 microporous membrane Substances 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 3
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 claims description 3
- 229920005654 Sephadex Polymers 0.000 claims description 2
- 239000012507 Sephadex™ Substances 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 238000001953 recrystallisation Methods 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 239000000499 gel Substances 0.000 claims 4
- 230000000694 effects Effects 0.000 abstract description 6
- 206010061218 Inflammation Diseases 0.000 abstract description 4
- 230000004054 inflammatory process Effects 0.000 abstract description 4
- 238000005516 engineering process Methods 0.000 abstract description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 abstract 1
- 239000002253 acid Substances 0.000 abstract 1
- 238000009472 formulation Methods 0.000 abstract 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 abstract 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 abstract 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 abstract 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 229910052708 sodium Inorganic materials 0.000 abstract 1
- 239000011734 sodium Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 20
- 238000005303 weighing Methods 0.000 description 6
- 230000035876 healing Effects 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 241000565359 Fraxinus chinensis Species 0.000 description 4
- 239000003513 alkali Substances 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 210000003560 epithelium corneal Anatomy 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000011555 rabbit model Methods 0.000 description 3
- 229910001753 sapphirine Inorganic materials 0.000 description 3
- 241001536358 Fraxinus Species 0.000 description 2
- 241000588214 Fraxinus chinensis subsp. rhynchophylla Species 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000010266 Sephadex chromatography Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
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- 239000000178 monomer Substances 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- GSDSWSVVBLHKDQ-UHFFFAOYSA-N 9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij]quinoline-6-carboxylic acid Chemical compound FC1=CC(C(C(C(O)=O)=C2)=O)=C3N2C(C)COC3=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-UHFFFAOYSA-N 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- GMRNMZUSKYJXGJ-UHFFFAOYSA-N Fraxetin Natural products C1=CC(=O)C(=O)C2=C1C=C(OC)C(O)=C2O GMRNMZUSKYJXGJ-UHFFFAOYSA-N 0.000 description 1
- 208000037319 Hepatitis infectious Diseases 0.000 description 1
- 206010027439 Metal poisoning Diseases 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 241000207834 Oleaceae Species 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- QNHQEUFMIKRNTB-UHFFFAOYSA-N aesculetin Natural products C1CC(=O)OC2=C1C=C(O)C(O)=C2 QNHQEUFMIKRNTB-UHFFFAOYSA-N 0.000 description 1
- GUAFOGOEJLSQBT-UHFFFAOYSA-N aesculetin dimethyl ether Natural products C1=CC(=O)OC2=C1C=C(OC)C(OC)=C2 GUAFOGOEJLSQBT-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000000146 antalgic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 208000030533 eye disease Diseases 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 229940020947 fluorescein sodium Drugs 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- HAVWRBANWNTOJX-UHFFFAOYSA-N fraxetin Chemical compound C1=CC(=O)OC2=C1C=C(OC)C(O)=C2O HAVWRBANWNTOJX-UHFFFAOYSA-N 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 208000005252 hepatitis A Diseases 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000008127 lead poisoning Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 description 1
- 229960001699 ofloxacin Drugs 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000007447 staining method Methods 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- GKCBAIGFKIBETG-UHFFFAOYSA-N tetracaine Chemical compound CCCCNC1=CC=C(C(=O)OCCN(C)C)C=C1 GKCBAIGFKIBETG-UHFFFAOYSA-N 0.000 description 1
- 229960002372 tetracaine Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
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- 239000003440 toxic substance Substances 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses aseculin ophthalmic gel and a production method and the application thereof; the aseculin ophthalmic gel provided by the invention is prepared by the following raw materials by mass parts: 4.8-5.2 parts of aseculin, 35-45 parts of boric acid, 380-420 parts of hydroxypropyl methyl cellulose, 180-220 parts of sodium alga acid, 23-27 parts of dimethyl fumarate and 49300-49350 parts of water; by adopting the formulation selection and preparation technology, the aseculin is produced into ophthalmic gel preparation for treating inflammation of eye part; the extraction method of the aseculin can lead the purity of the product to achieve 99.4 percent; the aseculin ophthalmic gel obtained in the invention is used for treating inflammation of eye part, the invention is firstly proposed and has high comfort degree and extract curative effect.
Description
Technical field
The present invention relates to a kind of Aseculin ophthalmic gel and production method thereof and application.
Background technology
Cortex Fraxini is dry branch skin or the dried bark of Oleaceae plant fraxinus rhynchophylla Hance (Fraxinus rhynchophylla Hance), Chinese ash (Fraxinus chinensis Roxb.), sharp leaf Chinese ash (Fraxinus szaboana Lingelsh.) or place post Chinese ash (Fraxinus stylosa Lingelsh.).Contain aseculin (C in the Cortex Fraxini
15H
16O
9).
Aseculin another name aesculin, esculetin glucoside have the effect of anti-inflammatory and antalgic, are used for the treatment of infectious hepatitis, acute biliary infection, lead poisoning, heat-clearing and toxic substances removing, vomiting and nausea uncomfortable in chest, cough due to lung-heat, tumor sore frequent fetal movement etc.
Summary of the invention
The purpose of this invention is to provide a kind of Aseculin ophthalmic gel and production method thereof and application.
Aseculin ophthalmic gel provided by the invention obtains with following feedstock production: 4.8-5.2 mass parts aseculin, 35-45 mass parts boric acid, 380-420 mass parts hydroxypropyl emthylcellulose, 180-220 mass parts sodium alginate, 23-27 mass parts dimethyl fumarate and 49300-49350 mass parts water.
Described Aseculin ophthalmic gel specifically can be and obtains with following feedstock production: 5 mass parts aseculin, 40 mass parts boric acid, 400 mass parts hydroxypropyl emthylcelluloses, 200 mass parts sodium alginates, 25 mass parts dimethyl fumarates and 49330 mass parts water.
The preparation method of described Aseculin ophthalmic gel is as follows: with described aseculin water dissolution, filtering and collecting filter liquid; With described boric acid, described hydroxypropyl emthylcellulose, described sodium alginate, described dimethyl fumarate water dissolution, obtain aseptic substrate after the sterilization; Described filtrate and described aseptic substrate are mixed, add entry, obtain Aseculin ophthalmic gel.
Described filtrate can be adopted 0.22 μ m filtering with microporous membrane.Sterilising conditions when preparing described aseptic substrate can be 121 ℃, 104kPa, 20min.
The preparation method of described aseculin specifically comprises the steps:
(1) Cortex Fraxini is carried out alcohol extraction, obtain crude extract;
(2) crude extract with step (1) mixes with ethyl acetate, abandons ethyl acetate layer after the layering, obtains extract;
(3) extract with step (2) carries out hydroxypropyl polydextran gel column chromatography, and the water eluting is collected aseculin solution and aseculin-fraxin mixed liquor;
(4) aseculin-fraxin mixed liquor with step (3) carries out hydroxypropyl polydextran gel column chromatography, uses the ethanol water eluting, collects aseculin solution;
(5) the aseculin solution with step (3) and step (4) carries out condensing crystallizing, collects crystal;
(6) with methanol the crystal of step (5) is carried out recrystallization, collect crystal;
(7) obtain aseculin after the crystal drying with step (6).
In the step (1), described alcohol extraction can be: Cortex Fraxini was extracted 1.5-2.5 hour for ethanol water 65-75 ℃ with 95% (quality percentage composition); The proportioning of described Cortex Fraxini and described 95% ethanol water is 1g Cortex Fraxini: 5-7mL 95% ethanol water; Described alcohol extraction is preferably: Ash Bark was extracted 2 hours for 70 ℃ with 95% ethanol water; The proportioning of described Cortex Fraxini and described 95% ethanol water is 1g Cortex Fraxini: 6mL 95% ethanol water.In the step (1), carry out described alcohol extraction after, can 55 ℃, the 0.1MPa evaporation, evaporation back volume obtains described crude extract for the 1/23-1/20 of evaporation front volume.
In the step (2), but described crude extract and described ethyl acetate equal-volume mix.
In the step (3), available distilled water eluting, flow velocity are 3.5mL/min.
In the step (4), available 20% (quality percentage composition) ethanol water eluting, flow velocity is 3.5mL/min.
In step (3) and the step (4), described hydroxypropyl sephadex column specifically can be hydroxypropyl polydextran gel Sephadex LH-20.
In the step (5), can be with 55 ℃ of described aseculin solution, 0.1MPa concentrating under reduced pressure, 0-4 ℃ of crystallize.
In the step (6), can filter and collect filtrate, 0-4 ℃ of crystallize with the crystal dissolve with methanol of step (5) in 55 ℃ of water-baths.
The present invention also protects a kind of medicine for the treatment of eye inflammation, and its active component is above arbitrary described Aseculin ophthalmic gel.
The present invention becomes eye-gel preparation by prescription screening and preparation technique with aseculin production, is used for the treatment of eye inflammation.The extracting method of aseculin provided by the invention can make the purity of product reach 99.4%.The Aseculin ophthalmic gel that the present invention obtains is used for the treatment of eye inflammation, for proposing this product comfort level height, determined curative effect first.
The specific embodiment
Following embodiment is convenient to understand better the present invention, but does not limit the present invention.Experimental technique among the following embodiment if no special instructions, is conventional method.Used test material among the following embodiment if no special instructions, is to buy from routine biochemistry reagent shop and obtains.
Cortex Fraxini: purchase in Bozhou City hundred full Pharmaceutical Co., Ltds; Boric acid: purchase in the poly-trade Co., Ltd of last sea fire; Hydroxypropyl emthylcellulose: purchase in last sea God's grace industry development company limited; Sodium alginate: purchase in Qingdao Mingyue Marine Alga Group Corp., Ltd.; Dimethyl fumarate: purchase in Shanghai Ou Le chemical industry company limited.
The preparation of embodiment 1, aseculin
1, alcohol extraction
Get 500g Cortex Fraxini coarse powder, add the ethanol 3000ml of 95% (quality percentage composition), 70 ℃ were extracted 2 hours, and were slightly carried filtrate after the filtration.Utilize the rotary evaporation in vacuo instrument will slightly propose filtrate decompression under 55 ℃, the condition of 0.1MPa and remove ethanol, obtain about 140ml crude extract to there not being the alcohol flavor.
2, extraction
In crude extract, add isopyknic ethyl acetate, keep water layer, abandon ethyl acetate layer, get extract.
3, chromatography
Adopt hydroxypropyl polydextran gel column chromatography, step is as follows:
(1) pretreatment
Hydroxypropyl polydextran gel Sephadex LH-20 (purchasing) in source, Shanghai leaf Bioisystech Co., Ltd, add ethanol, soaked 12 hours, after bubble is removed in stirring, pour in the chromatographic column, closure piston makes hydroxypropyl polydextran gel natural subsidence, when sedimentation exceeds 10cm, opens the piston ethanol elution, it is transparent to be washed till eluent, change distilled water and be eluted to nothing alcohol flavor, emit unnecessary distilled water in the post, treat sample.Chromatographic column diameter 3cm, glue height of bed 38cm.
(2) post separates
The extract of step 2 is joined on the pretreated hydroxypropyl sephadex chromatography post; With the distilled water eluting, flow velocity 3.5ml/min; Detect with uviol lamp, chromatographic column presents aeruginous fluorescence (aseculin-fraxin mixed liquor), sapphirine fluorescence (aseculin solution), dirty-green fluorescence (fraxetin solution) from bottom to up, collects respectively to present the eluent of aeruginous fluorescence and the eluent of sapphirine fluorescence; Obtain 340ml aseculin-fraxin mixed liquor, 670ml aseculin solution.
4, chromatography again
Aseculin-fraxin mixed liquor is concentrated into 200ml, goes up pretreated hydroxypropyl sephadex chromatography post again; Separate flow velocity 3.5ml/min with 20% (quality percentage composition) ethanol elution; Uviol lamp detects, and collects the eluent (aseculin solution) of sapphirine fluorescence, obtains 520ml aseculin solution.
5, crystallize
The aseculin solution that combining step 3 and step 4 obtain, 55 ℃, 0.1MPa are evaporated to 200ml, and 0-4 ℃ of crystallize filters and collect crystal.
6, recrystallize
Slowly add absolute methanol in 55 ℃ of water-baths in crystal, the limit edged stirs, and no longer reduces until crystal, filters and collects solution, and 0-4 ℃ of crystallize filters and collect crystal.
7, drying
100 ℃ of normal pressures of crystal are dry down, obtain 158mg aseculin monomer.
8, purity detecting
(1) test sample preparation: the lucifuge operation, precision takes by weighing the aseculin monomer (sample) that step 7 prepares, and per 100 μ g samples add 1ml water, obtain the solution first.
(2) reference substance preparation: the lucifuge operation, precision takes by weighing the aseculin standard substance (purchasing in Nat'l Pharmaceutical ﹠ Biological Products Control Institute) of dry constant weight, and per 100 μ g standard substance add 1ml water, obtain solution second.
(3) detect solution first and solution second respectively.Detection method: with octadecylsilane chemically bonded silica is filler; With acetonitrile-0.1% phosphoric acid solution (12: 88) is mobile phase; The detection wavelength is 334nm; Number of theoretical plate calculates by aseculin and is not less than 8000.Write down the chromatogram of sample and standard substance respectively.External standard method is calculated peak area, is the content of aseculin.Sample is the purity of sample with respect to the peak area ratio of standard substance.
Testing result shows that the purity of sample is 99.4%.
The preparation of embodiment 2, Aseculin ophthalmic gel
1, take by weighing the aseculin of 50mg embodiment 1 preparation, use the 200ml water dissolution, solution is collected filtrate through the 0.22um filtering with microporous membrane.
2, take by weighing 400mg boric acid, 4g hydroxypropyl emthylcellulose, 2g sodium alginate, 250mg dimethyl fumarate, use the 250ml water dissolution, get substrate; The substrate 20min that sterilizes under 121 ℃, 104kPa pressure obtains aseptic substrate;
3, the filtrate of step 1 and the aseptic substrate of step 2 are mixed, add water to 500g, mixing obtains Aseculin ophthalmic gel.
With Aseculin ophthalmic gel aluminum pipe fill, 5g/ props up.
Above-mentioned steps is all operated under aseptic condition, and employed vessel and aluminum pipe all carry out aseptic process in advance.
Embodiment 3, Aseculin ophthalmic gel are to the curative effect of eye inflammation
One, makes the model of alkali burned rabbit model
(Liaoning Medical University's Experimental Animal Center, the quality certification number: No. 038, distant real kinoplaszm card word), body weight is 2.5～3.5Kg, male and female dual-purpose, healthy no oculopathy to adopt Japanese white big ear rabbit.With 1% tetracaine topical anesthesia; With diameter is that the filter paper (being soaked with 0.5mol/L NaOH solution) of 8mm places anterior corneal surface central authorities; Take off filter paper behind the 1min; Wash cornea 2min continuously with sterile saline; Obtain the model of alkali burned rabbit model.
Two, Aseculin ophthalmic gel is to the curative effect of eye inflammation
The model of alkali burned rabbit model is divided into 3 groups at random, 10 every group, carry out administration respectively, specific as follows:
First group (blank group): drip normal saline, four times a day, splashes into and suffers from eye by each one;
Second group (positive controls): drip ofloxacin eye drops (Cangzhou Guangming Pharmaceutical Co., Ltd), four times a day, splashes into and suffers from eye by each one;
The 3rd group (treatment group): drip the Aseculin ophthalmic gel of embodiment 2 preparations, four times a day, splashes into and suffers from eye by each one.
2 weeks of successive administration, observe the inflammation that test sample causes alkali burn and the relief capabilities of edema, calculate the corneal epithelium healing rate, to estimate the therapeutic effect of each test sample.
Calculate the corneal epithelium healing rate and adopt fluorescent staining method: with one of 20% fluorescein sodium sterile solution, splash in the rabbit conjunctival sac, make its passive 5min of closing one's eyes, stop a moment then slightly, if ophthalmic still has more dyestuff, available normal saline solution flushing is removed it, carried out slit lamp examination in 0,1,2,3,5,7,10,14 day respectively at hindering the back, fluorescent staining is taken a picture, and photo machine image analysis system is as calculated handled, and calculates its corneal epithelium healing rate.。
Corneal epithelial wound area (stained area) sees Table 1.Corneal epithelial cell healing rate (%) sees Table 2.
Table 1 corneal epithelial wound area (mm
2, X ± SD)
| Group | First group | Second group | The 3rd group |
| 0 day | ??50.37±0.16 | ??50.12±0.15 | ??49.93±0.26 |
| The 1st day | ??47.62±1.44 | ??36.62±1.42 | ??35.73±2.37 |
| The 2nd day | ??41.49±1.83 | ??36.48±1.87 | ??35.71±2.5 |
| The 3rd day | ??39.47±3.5 | ??35.87±3.3 | ??33.29±1.6 |
| The 5th day | ??39.17±3.21 | ??33.17±3.19 | ??28.77±1.25 |
| The 7th day | ??35.5±2.5 | ??32.5±2.5 | ??25.68±3.81 |
| The 10th day | ??34.57±3.32 | ??24.67±1.91 | ??17.95±1.34 |
| The 14th day | ??31.69±1.91 | ??22.57±3.32 | ??16.67±0.44 |
Table 2 corneal epithelial cell healing rate (%)
| Group | First group | Second group | The 3rd group |
| 0 day | ??0 | ??0 | ??0 |
| The 1st day | ??5.5 | ??26.9 | ??28.4 |
| The 2nd day | ??17.6 | ??27.2 | ??28.4 |
| The 3rd day | ??21.6 | ??28.4 | ??33.3 |
| The 5th day | ??22.2 | ??33.8 | ??42.4 |
| The 7th day | ??29.5 | ??35.2 | ??48.6 |
| The 10th day | ??31.4 | ??50.8 | ??64.0 |
| The 14th day | ??37.1 | ??54.0 | ??66.6 |
Experimental result shows that positive controls and treatment group be amelioration of inflammation effectively, promotes wound healing, and the effect of treatment group is best.
The eye comfort level of embodiment 4, Aseculin ophthalmic gel and availability are investigated
1, take by weighing the aseculin of 50mg embodiment 1 preparation, use the 200ml water dissolution, solution is collected filtrate through the 0.22um filtering with microporous membrane.
2, take by weighing 400mg boric acid, hydroxypropyl emthylcellulose (consumption sees Table 3), sodium alginate (consumption sees Table 4), 250mg dimethyl fumarate, use the 250ml water dissolution, get substrate; The substrate 20min that sterilizes under 121 ℃, 104kPa pressure obtains aseptic substrate;
3, the filtrate of step 1 and the aseptic substrate of step 2 are mixed, add water to 500g, mixing obtains Aseculin ophthalmic gel.
The addition of hydroxypropyl emthylcellulose and sodium alginate in four kinds of samples of table 3.
| Sample number | Hydroxypropyl emthylcellulose | Sodium alginate |
| ??1 | ??6g | ??0g |
| ??2 | ??4g | ??2g |
| ??3 | ??2g | ??4g |
| ??4 | ??0 | ??6g |
Investigate the comfort level of four kinds of samples.Comfort level evaluation criteria (after the administration in 15 seconds number of winks): 0-4 time: ++ ++; 4-6 time: +++; 6-8 time: ++; More than 8 times :+.The results are shown in Table 4.
The comfort level of four kinds of samples of table 4
Wherein sample 2 comfort levels are best.
Claims (10)
1. an Aseculin ophthalmic gel obtains with following feedstock production: 4.8-5.2 mass parts aseculin, 35-45 mass parts boric acid, 380-420 mass parts hydroxypropyl emthylcellulose, 180-220 mass parts sodium alginate, 23-27 mass parts dimethyl fumarate and 49300-49350 mass parts water.
2. Aseculin ophthalmic gel as claimed in claim 1 is characterized in that: obtain with following feedstock production: 5 mass parts aseculin, 40 mass parts boric acid, 400 mass parts hydroxypropyl emthylcelluloses, 200 mass parts sodium alginates, 25 mass parts dimethyl fumarates and 49330 mass parts water.
3. Aseculin ophthalmic gel as claimed in claim 1 or 2 is characterized in that: prepare with the following method: with described aseculin water dissolution, filtering and collecting filter liquid; With described boric acid, described hydroxypropyl emthylcellulose, described sodium alginate, described dimethyl fumarate water dissolution, obtain aseptic substrate after the sterilization; Described filtrate and described aseptic substrate are mixed, add entry, obtain Aseculin ophthalmic gel.
4. Aseculin ophthalmic gel as claimed in claim 3 is characterized in that: described filtrate is adopted 0.22 μ m filtering with microporous membrane; Sterilising conditions when preparing described aseptic substrate is 121 ℃, 104kPa, 20min.
5. as arbitrary described Aseculin ophthalmic gel in the claim 1 to 4, it is characterized in that: the preparation method of described aseculin comprises the steps:
(1) Cortex Fraxini is carried out alcohol extraction, obtain crude extract;
(2) crude extract with step (1) mixes with ethyl acetate, abandons ethyl acetate layer after the layering, obtains extract;
(3) extract with step (2) carries out hydroxypropyl polydextran gel column chromatography, and the water eluting is collected aseculin solution and aseculin-fraxin mixed liquor;
(4) aseculin-fraxin mixed liquor with step (3) carries out hydroxypropyl polydextran gel column chromatography, uses the ethanol water eluting, collects aseculin solution;
(5) the aseculin solution with step (3) and step (4) carries out condensing crystallizing, collects crystal;
(6) with methanol the crystal of step (5) is carried out recrystallization, collect crystal;
(7) obtain aseculin after the crystal drying with step (6).
6. Aseculin ophthalmic gel as claimed in claim 5 is characterized in that: in the step (1), described alcohol extraction is: Cortex Fraxini was extracted 1.5-2.5 hour for ethanol water 65-75 ℃ with 95% (quality percentage composition); The proportioning of described Cortex Fraxini and described 95% ethanol water is 1g Cortex Fraxini: 5-7mL 95% ethanol water;
Described alcohol extraction is preferably: Ash Bark was extracted 2 hours for 70 ℃ with 95% ethanol water; The proportioning of described Cortex Fraxini and described 95% ethanol water is 1g Cortex Fraxini: 6mL 95% ethanol water.
7. as claim 5 or 6 described Aseculin ophthalmic gels, it is characterized in that: in the step (1), carry out described alcohol extraction after, 55 ℃, 0.1MPa evaporation, evaporation back volume obtains described crude extract for the 1/23-1/20 of evaporation front volume.
8. as arbitrary described Aseculin ophthalmic gel in the claim 5 to 7, it is characterized in that: in the step (2), described crude extract and described ethyl acetate equal-volume mix; In the step (5), with 55 ℃ of described aseculin solution, 0.1MPa concentrating under reduced pressure, 0-4 ℃ of crystallize; In the step (6), with the crystal dissolve with methanol of step (5), filter and collect filtrate, 0-4 ℃ of crystallize in 55 ℃ of water-baths.
9. as arbitrary described Aseculin ophthalmic gel in the claim 5 to 8, it is characterized in that: in the step (3), use the distilled water eluting, flow velocity is 3.5mL/min; In the step (4), with 20% (quality percentage composition) ethanol water eluting, flow velocity is 3.5mL/min; In step (3) and the step (4), described hydroxypropyl sephadex column is hydroxypropyl polydextran gel Sephadex LH-20.
10. medicine for the treatment of eye inflammation, its active component is arbitrary described Aseculin ophthalmic gel in claim 1 or 9.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102091132A (en) * | 2011-01-25 | 2011-06-15 | 西南民族大学 | Method for detecting esculin, aesculetin, fraxin and fraxetin in cortex fraxini or extract thereof |
| CN104844670A (en) * | 2014-02-18 | 2015-08-19 | 中国医学科学院药物研究所 | Esculin crystal form B substance, preparation method, composition, and uses thereof |
| CN109134562A (en) * | 2018-10-26 | 2019-01-04 | 宁县恒瑞康生物科技有限公司 | A kind of aesculin purification process |
| CN110538137A (en) * | 2019-09-30 | 2019-12-06 | 辽宁大学 | aesculin nano suspension gel and preparation method and application thereof |
| CN113143862A (en) * | 2021-06-23 | 2021-07-23 | 顾凌雯 | Dimethyl fumarate eye drops, preparation method thereof and application of dimethyl fumarate eye drops as fungal keratitis treatment medicine |
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2009
- 2009-12-08 CN CN2009102421095A patent/CN101716133B/en active Active
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102091132A (en) * | 2011-01-25 | 2011-06-15 | 西南民族大学 | Method for detecting esculin, aesculetin, fraxin and fraxetin in cortex fraxini or extract thereof |
| CN102091132B (en) * | 2011-01-25 | 2013-05-01 | 西南民族大学 | Method for detecting esculin, aesculetin, fraxin and fraxetin in cortex fraxini or extract thereof |
| CN104844670A (en) * | 2014-02-18 | 2015-08-19 | 中国医学科学院药物研究所 | Esculin crystal form B substance, preparation method, composition, and uses thereof |
| CN109134562A (en) * | 2018-10-26 | 2019-01-04 | 宁县恒瑞康生物科技有限公司 | A kind of aesculin purification process |
| CN110538137A (en) * | 2019-09-30 | 2019-12-06 | 辽宁大学 | aesculin nano suspension gel and preparation method and application thereof |
| CN113143862A (en) * | 2021-06-23 | 2021-07-23 | 顾凌雯 | Dimethyl fumarate eye drops, preparation method thereof and application of dimethyl fumarate eye drops as fungal keratitis treatment medicine |
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|---|---|
| CN101716133B (en) | 2011-10-05 |
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