CN100528224C - Slow release microphere for injection containing interferon alpha-1b and its preparation method - Google Patents
Slow release microphere for injection containing interferon alpha-1b and its preparation method Download PDFInfo
- Publication number
- CN100528224C CN100528224C CNB2004100826944A CN200410082694A CN100528224C CN 100528224 C CN100528224 C CN 100528224C CN B2004100826944 A CNB2004100826944 A CN B2004100826944A CN 200410082694 A CN200410082694 A CN 200410082694A CN 100528224 C CN100528224 C CN 100528224C
- Authority
- CN
- China
- Prior art keywords
- alfacon
- microsphere
- slow release
- injection
- release microphere
- 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.)
- Expired - Fee Related
Links
- 239000007924 injection Substances 0.000 title claims abstract description 17
- 238000002347 injection Methods 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title abstract description 14
- 101000959820 Homo sapiens Interferon alpha-1/13 Proteins 0.000 title abstract 2
- 102100040019 Interferon alpha-1/13 Human genes 0.000 title abstract 2
- 238000000034 method Methods 0.000 claims abstract description 24
- 239000004005 microsphere Substances 0.000 claims description 46
- 239000000243 solution Substances 0.000 claims description 19
- 229920003168 pharmaceutical polymer Polymers 0.000 claims description 13
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 11
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 11
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 239000002202 Polyethylene glycol Substances 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 229920001577 copolymer Polymers 0.000 claims description 8
- 229920001223 polyethylene glycol Polymers 0.000 claims description 8
- 239000000741 silica gel Substances 0.000 claims description 8
- 229910002027 silica gel Inorganic materials 0.000 claims description 8
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 claims description 6
- 241001597008 Nomeidae Species 0.000 claims description 6
- 239000003381 stabilizer Substances 0.000 claims description 6
- BPMFZUMJYQTVII-UHFFFAOYSA-N alpha-guanidinoacetic acid Natural products NC(=N)NCC(O)=O BPMFZUMJYQTVII-UHFFFAOYSA-N 0.000 claims description 5
- 239000002502 liposome Substances 0.000 claims description 5
- -1 polypropylene Polymers 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- 239000003208 petroleum Substances 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 108010010803 Gelatin Proteins 0.000 claims description 3
- 239000008273 gelatin Substances 0.000 claims description 3
- 229920000159 gelatin Polymers 0.000 claims description 3
- 235000019322 gelatine Nutrition 0.000 claims description 3
- 235000011852 gelatine desserts Nutrition 0.000 claims description 3
- 229960004275 glycolic acid Drugs 0.000 claims description 3
- 239000002105 nanoparticle Substances 0.000 claims description 3
- 229920000747 poly(lactic acid) Polymers 0.000 claims description 3
- 239000004626 polylactic acid Substances 0.000 claims description 3
- 210000000582 semen Anatomy 0.000 claims description 3
- 102000009027 Albumins Human genes 0.000 claims description 2
- 108010088751 Albumins Proteins 0.000 claims description 2
- 229920001503 Glucan Polymers 0.000 claims description 2
- 229920002732 Polyanhydride Chemical group 0.000 claims description 2
- 229920000954 Polyglycolide Polymers 0.000 claims description 2
- 239000004743 Polypropylene Substances 0.000 claims description 2
- 150000002148 esters Chemical group 0.000 claims description 2
- 239000004310 lactic acid Substances 0.000 claims description 2
- 235000014655 lactic acid Nutrition 0.000 claims description 2
- TWNIBLMWSKIRAT-VFUOTHLCSA-N levoglucosan Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@H]2CO[C@@H]1O2 TWNIBLMWSKIRAT-VFUOTHLCSA-N 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000004633 polyglycolic acid Substances 0.000 claims description 2
- 239000002861 polymer material Substances 0.000 claims description 2
- 229920001155 polypropylene Polymers 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 6
- 238000001694 spray drying Methods 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 15
- 238000003756 stirring Methods 0.000 description 15
- 238000004108 freeze drying Methods 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 8
- 238000013268 sustained release Methods 0.000 description 7
- 239000012730 sustained-release form Substances 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 239000006101 laboratory sample Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical class OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 229930195725 Mannitol Natural products 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000001033 granulometry Methods 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Chemical class OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical class OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 238000012449 Kunming mouse Methods 0.000 description 2
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical group C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 2
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Chemical class O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical class O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005354 coacervation Methods 0.000 description 2
- 239000006184 cosolvent Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000004945 emulsification Methods 0.000 description 2
- 230000001804 emulsifying effect Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000005304 joining Methods 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000008176 lyophilized powder Substances 0.000 description 2
- 238000002663 nebulization Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000012875 nonionic emulsifier Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 229960000502 poloxamer Drugs 0.000 description 2
- 229920001983 poloxamer Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 239000003440 toxic substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- RKDVKSZUMVYZHH-UHFFFAOYSA-N 1,4-dioxane-2,5-dione Chemical compound O=C1COC(=O)CO1 RKDVKSZUMVYZHH-UHFFFAOYSA-N 0.000 description 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 210000000026 apposition eye Anatomy 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000006189 buccal tablet Substances 0.000 description 1
- 229940046011 buccal tablet Drugs 0.000 description 1
- 239000012531 culture fluid Substances 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000010556 emulsion polymerization method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000008394 flocculating agent Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- JJTUDXZGHPGLLC-UHFFFAOYSA-N lactide Chemical compound CC1OC(=O)C(C)OC1=O JJTUDXZGHPGLLC-UHFFFAOYSA-N 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 229960003511 macrogol Drugs 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000011806 microball Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- OHDXDNUPVVYWOV-UHFFFAOYSA-N n-methyl-1-(2-naphthalen-1-ylsulfanylphenyl)methanamine Chemical compound CNCC1=CC=CC=C1SC1=CC=CC2=CC=CC=C12 OHDXDNUPVVYWOV-UHFFFAOYSA-N 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000010603 pastilles Nutrition 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000151 polyglycol Polymers 0.000 description 1
- 239000010695 polyglycol Substances 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 229920002545 silicone oil Polymers 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to slow release microballoons for injection of alpha-1b and the process for preparation, wherein the slow release microballoons comprise (by weight ratio), 0.1-10% (w/w) of interferon alpha-1b or its derivatives, 50-99.9% (w/w) of biodegradable and biologically compatible medicinal macromolecular material with a molecular weight between 5000-70000 Daltons. The microballoons can be prepared through W/O/W method, S/O/O method and spray-drying method.
Description
Technical field
The present invention relates to slow release microphere for injection of Alfacon-1 b and preparation method thereof.
Background technology
Find by retrieval, the preparation of Alfacon-1 b or derivatives thereof or salt has buccal tablet, solution, injection, patch, ointment etc., these dosage forms or since half-life of Alfacon-1 b shorter, cause eliminating very fast, or owing to non local use makes medicine on the low side and the whole body toxic and side effects is big at partial dose, clinical practice is restricted.
Summary of the invention
The technical problem to be solved in the present invention, purpose promptly of the present invention provides a kind of slow release microphere for injection of Alfacon-1 b or derivatives thereof.
Through research for a long time, the present inventor finds, to account for Alfacon-1 b or derivatives thereof and 50～99.9% (w/w) molecular weight of 0.1～10% (w/w) of microsphere weight 5,000～70, the pharmaceutical polymers of the biodegradable and bio-compatible between 000 dalton is made slow release microphere for injection by appropriate methodology, can realize above-mentioned purpose of the present invention.
Therefore, the invention provides a kind of slow release microphere for injection, it is characterized in that it comprises in microsphere weight, 0.1 Alfacon-1 b or derivatives thereof and 50～99.9% (w/w) molecular weight of～10% (w/w) is 5,000～70, biodegradable between 000 dalton and have the pharmaceutical polymers of biocompatibility, the mean diameter of described microsphere is between 10～100 μ m.
Slow release microphere for injection of the present invention can prepare by the either party's method in the following method.
Multi-emulsion method, it is characterized in that after an amount of Alfacon-1 b or derivatives thereof and stabilizing agent dissolving or being suspended in the aqueous solution that contains nonionic emulsifier, join in the organic solution that contains described pharmaceutical polymers, after treatment, join the outer aqueous phase that contains nonionic emulsifier again and form microsphere.
No water law, it is characterized in that the micropowder of an amount of Alfacon-1 b or derivatives thereof and stabilizing agent thereof is joined in the organic solvent of pbz polymer material, behind the ultrasonic suspendible of water-bath, join again in a certain amount of Oleum Gossypii semen, make it be solidified into microsphere with the decentralized photo extraction, add light petroleum ether then, make the microsphere completion of cure.
The cold nebulization extraction method, it is characterized in that an amount of Alfacon-1 b or derivatives thereof powder suspension in being dissolved with the organic solvent of macromolecular material, form droplet by spraying, freezing in liquid nitrogen, make with low temperature organic cosolvent extracting dissolve polymer.
The phase coacervation after it is characterized in that an amount of Alfacon-1 b or derivatives thereof is scattered in suitable macromolecule material solution, adds flocculating agent in this solution, the macromolecular material dissolubility is reduced and separate out, and forms good microsphere.
Sustained-release micro-spheres of the present invention also can be by after being prepared into liposome with Alfacon-1 b or derivatives thereof earlier, and the described pharmaceutical polymers that reuse is fit to is rolled into the method preparation of microsphere.
Sustained-release micro-spheres of the present invention also can be by after being prepared into nanoparticle with Alfacon-1 b or derivatives thereof earlier, and the described pharmaceutical polymers that reuse is fit to is rolled into the method preparation of microsphere.
Sustained-release micro-spheres of the present invention also can be by joining Alfacon-1 b or derivatives thereof solution in the micropowder silica gel, and after fully being adsorbed by micropowder silica gel, the described pharmaceutical polymers that reuse is fit to is rolled into the method preparation of microsphere.
Alfacon-1 b derivant of the present invention comprise various Alfacon-1 b salt, PEGization, methylate and the derivant of the various Alfacon-1 b that modify with other group.
Pharmaceutical polymers of the present invention is a gelatin, albumin, polylactide-co-glycolide, polylactic acid, polyglycolic acid, poly--the 3-butyric ester, poly-adjacent ester, polyanhydride, poly butyric ester-hydroxyl pentanoate copolymer, the polypropylene glucosan, polylactone, polyvinyl alcohol (PVA), Polyethylene Glycol (PEG), hydroxyacetic acid, polylactic acid-polyglycol, polylactic acid-glycolic guanidine-acetic acid copolymer (PLGA), a kind of in polyglycolic acid-Polyethylene Glycol or two or more mixture wherein.
Be preferably polylactic acid, polylactide-co-glycolide, polylactic acid-glycolic guanidine-acetic acid copolymer or poly butyric ester-hydroxyl pentanoate copolymer.
More preferably molecular weight is 5,000～70, polylactic acid-glycolic guanidine-acetic acid copolymer between 000 dalton, wherein the ratio of lactic acid and hydroxyacetic acid is 20: 80～80: 20, perhaps molecular weight is 5, polylactide-co-glycolide between 000～70,000 dalton, wherein the polymerization ratio of lactide and Acetic acid, hydroxy-, bimol. cyclic ester is 20: 80～80: 20.
Injectable microsphere of the present invention also can comprise pharmaceutically acceptable protein stabiliser, surfactant and other additives etc.
Described protein stabiliser is selected from HSA, mannitol, trehalose, EDTA, Zn salt, Mg (OH)
2, CaCO
3, ZnCO
3Deng.
Described surfactant is selected from poloxamer, lecithin, Macrogol 2000 or Polyethylene Glycol 5000 etc.
Described other additives are selected from micropowder silica gel, carry positive ionic material such as aminoacid etc.
Microball preparation envelop rate of the present invention is greater than 75%, and sustainable release reduced the medication number of times about 15 days, 30 days or 60 days, improved bioavailability, reduced systemic toxic side effect, increased patient compliance.
The specific embodiment
Further specify the preparation method of the slow release microphere for injection of Alfacon-1 b or derivatives thereof of the present invention by the following examples.
Embodiment 1: multi-emulsion method prepares recombinantinterferon-1b sustained-release micro-spheres
(about 5mg is with Zn with a certain amount of Alfacon-1 b
2+In conjunction with the back) be dissolved in the 0.5ml20mg/ml gelatin solution and (contain EDTA, 10% mannitol or trehalose, Mg (OH)
2, CaCO
3Or ZnCO
3Micropowder etc.) in, after 5000rpm stirs 30s, join among the dichloromethane solution 1.25ml that contains 150mgPLGA (molecular weight is 5000～70,000), under the rotating speed of 5000rpm, stirred 2 minutes, joining 20ml then contains in the aqueous solution of 6%PVA/PEG, with 200rpm speed magnetic agitation 5 minutes, and then join in the 500ml aqueous solution that contains 2% PVA/PEG, stir and carry out solvent evaporates, washing, centrifugal, lyophilization then collected promptly.
Embodiment 2: anhydrous legal system is equipped with recombinantinterferon-1b sustained-release micro-spheres
The about 150mg of PLGA is dissolved among acetonitrile or the ethyl acetate 1ml, then to the micropowder that wherein adds Alfacon-1 b or derivatives thereof and stabilizing agent thereof, behind the ultrasonic suspendible of water-bath, join in the Oleum Gossypii semen (containing lecithin or poloxamer etc.) of 20-30ml as emulsifying agent, stirring and emulsifying, the acetonitrile solution that constantly extracts in the emulsion droplet with decentralized photo makes it be solidified into microsphere, add light petroleum ether (boiling point 60-90 ℃) then, continue to stir and to make the microsphere completion of cure, centrifugal, petroleum ether, lyophilization are promptly.
Embodiment 3: the cold nebulization extraction method prepares recombinantinterferon-1b sustained-release micro-spheres
Alfacon-1 b or derivatives thereof powder suspension is in being dissolved with the organic solvent of macromolecular material, freezing in liquid nitrogen by spraying to such an extent that method forms droplet, get solvent promptly with a low temperature organic cosolvent (for example ethanol) extracting dissolve polymer.
Embodiment 4: be rolled into microsphere again after making liposome earlier
Adopt film dispersion method or freeze-drying etc. to prepare Alfacon-1 b liposome, then the liposome powder after an amount of lyophilization is joined among the dichloromethane solution 1ml of 100mg/ml PLGA, behind ultrasonic/stirring suspendible, join among the aqueous solution 15ml of PVA, join in the dilution PVA solution of 200ml after stirring 5min with the rotating speed of about 2000rpm, after continue stirring 4h with the slow-speed of revolution, washing, centrifugal, lyophilization are promptly.
Embodiment 5: be rolled into microsphere again after making nanoparticle earlier
After adopting natural polymer method and emulsion polymerization method to be prepared into nano-granule freeze-dried powder, join among the dichloromethane solution 1ml of 100mg/ml PLGA, behind ultrasonic/stirring suspendible, join among the aqueous solution 15ml of PVA, join in the dilution PVA solution of 200ml after stirring 5min with the rotating speed of about 2000rpm, after continue stirring 4h with the slow-speed of revolution, washing, centrifugal, lyophilization are promptly.
Embodiment 6: be rolled into microsphere again after micropowder silica gel absorption
Earlier Alfacon-1 b buffer is joined in the micropowder silica gel; after fully being adsorbed by micropowder silica gel; add freeze drying protectants such as mannitol; after lyophilization is prepared into lyophilized powder, join in the dichloromethane solution of 100mg/ml PLGA, join among the PVA aqueous solution 15ml behind ultrasonic/stirring suspendible; join after the rotating speed stirring and emulsifying with about 2000rpm in the PVA solution of dilution; continue to stir 4h with the slow-speed of revolution, solvent flashing, after washing, centrifugal, the lyophilization promptly.
Embodiment 7: the phase coacervation prepares microsphere
Alfacon-1 b lyophilized powder or Alfacon-1 b solution are joined in the PLGA dichloromethane solution of 150mg/ml in right amount, and behind the rotating speed stirring 1min with about 2000rpm, the reduction rotating speed is 200rpm, and injects silicone oil.After being separated, suspension is changed in the hexane of 500ml, after 20 ℃ of rotating speeds with about 300rpm of constant temperature stir 30min, collect microsphere, the 500ml hexane wash, lyophilization is promptly.
The mensuration of microspherulite diameter
Test specimen: according to the microsphere of the embodiment of the invention 1,2,3,4,5,6,7 methods preparation.
Experimental apparatus: the optical microscope or the laser granulometry of band graduated eyepiece.
Experimental technique:
Light microscope determining: will observe down with last embodiment microsphere at microscope (laser counter) behind the microsphere powder homodisperse, and write down microspherulite diameter, the microsphere sum is no less than 200, and data are carried out statistical disposition, investigates the mean diameter and the particle size distribution situation of microsphere.
Laser granulometry is measured: take by weighing suitable laboratory sample, use laser granulometry to measure.
The mensuration of microsphere drug loading and envelop rate
Test specimen: according to the microsphere of the embodiment of the invention 1,2,3,4,5 methods preparation.
Experiment reagent: BCA determining the protein quantity test kit, NaOH, SDS
Experimental apparatus: ultraviolet spectrophotometer, vortex instrument, centrifuge
Experimental technique: precision takes by weighing the about 10mg of laboratory sample, adds in the mixed solution of 5.0ml 0.1MNaOH/0.5SDS (w/v), and 37 ℃ of water-baths vibration 24h dissolve microsphere fully, adds the about 280 μ l of 2MHCL and transfers pH about 7.0, carries out BCA method determining the protein quantity.
Microsphere prominent released the mensuration with release profiles
Laboratory sample: according to the microsphere of the embodiment of the invention 1,2,3 described method preparations.
Experiment reagent: pH is 7.4 phosphate buffer (contain 0.02% Hydrazoic acid,sodium salt as antibacterial, the 0.02%F-68 wetting agent is as release medium)
Experimental apparatus: constant temperature shaker, centrifuge, HPLC instrument.
Experiment condition: temperature: 37 ℃, rotating speed: rpm.
Experimental technique: precision takes by weighing pastille microsphere 10-30mg and puts in the 10ml centrifuge tube, adds 1.5ml10mM pH 7.4 buffer, places the water bath with thermostatic control shaking table, carries out the release in vitro degree of microsphere and measure under 100rpm hunting speed, 37 ℃ ± 0.5 ℃ temperature conditions.
Sampling method: take out centrifuge tube at 2h, 1d, 2d respectively, in the centrifugal 10min of 2000rpm,, took a sample once with above operational approach every 3-4 days later on, and measure the content of IFN with the BCA method with the release medium that whole release medium are taken out and more renewed.
Microsphere is measured at the intravital sustained release property of mice
Laboratory sample: according to the microsphere of the embodiment of the invention 1,2,3 described method preparations.
Laboratory animal: Kunming mouse
Experimental technique: get healthy Kunming mouse, after the random packet, vein gives the microspheres solution of rIFN α, and dosage is 1.5 * 10
4KU rIFN α/kg, 0,1,2,5,8,10,12,15 day eye socket is got blood after administration, and blood sample is got supernatant and is measured wherein IFN-α biological activity behind the centrifugal 15min of 3000rpm.
The bioactive assay method of Alfacon-1 b
Laboratory sample: plasma sample.
Experimental apparatus: freezer dryer, centrifuge.
Experimental technique: the bioactive mensuration of Alfacon-1 b: WISH cell through 0.25% trypsinization, is made into 2.5 * 10 with complete culture solution
5-3.5 * 10
5Individual cell/ml is inoculated in 96 porocyte culture plates, every hole 100 μ l.37 ℃, 5%CO
2Cultivated 4-6 hour under the condition, supernatant discarded, to disturb plain α-1b sample solution and standard solution and carry out adding 1-12 hole (sample solution that concentration is high dilutes in advance) from high to low by concentration behind the doubling dilution, set up the contrast of cell contrast and virus control and sample blank simultaneously.Similarity condition was cultivated 18-24 hour, and supernatant discarded adds VSV counteracting toxic substances culture fluid (dosage 100TCID50) 100 μ l, counteracting toxic substances 24 hours, and microscopy cell 50% pathological changes point under the inverted microscope, calculating is disturbed plain α-1b and is tired.
Claims (7)
1. slow release microphere for injection, it is characterized in that it comprises in microsphere weight, 0.1 the Alfacon-1 b of～10% (w/w) or its derivant and 50～99.9% (w/w) molecular weight of being selected from salt, the PEGization Alfacon-1 b of Alfacon-1 b or the Alfacon-1 b that methylates are 5,000～70, biodegradable between 000 dalton and have the pharmaceutical polymers of biocompatibility, the mean diameter of described microsphere is between 10～100 μ m.
2. according to the slow release microphere for injection of claim 1, wherein said pharmaceutical polymers is selected from a kind of in gelatin, albumin, polylactic acid, polyglycolic acid, poly--the 3-butyric ester, poly-adjacent ester, polyanhydride, poly butyric ester-hydroxyl pentanoate copolymer, polypropylene glucosan, polylactone, polyvinyl alcohol, Polyethylene Glycol, polylactic acid-glycolic guanidine-acetic acid copolymer and the polyglycolic acid-Polyethylene Glycol or two or more mixture wherein.
3. according to the slow release microphere for injection of claim 2, wherein said pharmaceutical polymers is a polylactic acid-glycolic guanidine-acetic acid copolymer, and wherein molecular weight is 5,000～70, and between 000 dalton, the ratio of lactic acid and hydroxyacetic acid is 20: 80～80: 20.
4. one kind prepares any one the method for slow release microphere for injection of claim 1 to 3, it is characterized in that the micropowder of an amount of Alfacon-1 b or described its derivant and stabilizing agent thereof is joined in the organic solvent of pbz polymer material, behind the ultrasonic suspendible of water-bath, join again in a certain amount of Oleum Gossypii semen, make it be solidified into microsphere with the decentralized photo extraction, add light petroleum ether then, make the microsphere completion of cure.
5. one kind prepares any one the method for slow release microphere for injection of claim 1 to 3, and it is characterized in that earlier will an amount of Alfacon-1 b or after described its derivant was prepared into liposome, the suitable described pharmaceutical polymers of reuse was rolled into microsphere.
6. one kind prepares any one the method for slow release microphere for injection of claim 1 to 3, and it is characterized in that earlier will an amount of Alfacon-1 b or after described its derivant was prepared into nanoparticle, the suitable described pharmaceutical polymers of reuse was rolled into microsphere.
7. one kind prepares any one the method for slow release microphere for injection of claim 1 to 3, it is characterized in that an amount of Alfacon-1 b or described its derivative solution are joined in the micropowder silica gel, after fully being adsorbed by micropowder silica gel, the described pharmaceutical polymers that reuse is fit to is rolled into microsphere.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100826944A CN100528224C (en) | 2004-09-27 | 2004-09-27 | Slow release microphere for injection containing interferon alpha-1b and its preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100826944A CN100528224C (en) | 2004-09-27 | 2004-09-27 | Slow release microphere for injection containing interferon alpha-1b and its preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1754572A CN1754572A (en) | 2006-04-05 |
| CN100528224C true CN100528224C (en) | 2009-08-19 |
Family
ID=36688303
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100826944A Expired - Fee Related CN100528224C (en) | 2004-09-27 | 2004-09-27 | Slow release microphere for injection containing interferon alpha-1b and its preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100528224C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| UY33517A (en) * | 2010-07-19 | 2012-02-29 | Astrazeneca Ab | Pharmaceutical depot for 5-fluoro-2 - [[(1S) -1- (5-fluoro-2-pyridyl) ethyl] amino] -6 - [(5-isopropoxy-1H-pyrazol-3-yl) amino] pyridin -3-carbonitrile ?. |
| CN105878191B (en) * | 2016-04-26 | 2021-01-22 | 广州帝奇医药技术有限公司 | Preparation method of sustained-release microparticles, prepared sustained-release microparticles and application thereof |
| CN105878190B (en) * | 2016-04-26 | 2021-01-22 | 广州帝奇医药技术有限公司 | Preparation method of sustained-release microparticles, prepared sustained-release microparticles and application thereof |
| EP4065086A1 (en) | 2019-11-25 | 2022-10-05 | Regeneron Pharmaceuticals, Inc. | Sustained release formulations using non-aqueous emulsions |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1154653A (en) * | 1994-07-25 | 1997-07-16 | 阿尔克姆斯控制治疗公司 | Controlled release of metal cation-stabilized interferon |
| WO1999020301A1 (en) * | 1997-10-18 | 1999-04-29 | Boehringer Ingelheim International Gmbh | Tumour vaccine |
-
2004
- 2004-09-27 CN CNB2004100826944A patent/CN100528224C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1154653A (en) * | 1994-07-25 | 1997-07-16 | 阿尔克姆斯控制治疗公司 | Controlled release of metal cation-stabilized interferon |
| US6379701B1 (en) * | 1994-07-25 | 2002-04-30 | Alkermes Controlled Therapeutics, Inc. | Controlled release of metal cation-stabilized interferon |
| WO1999020301A1 (en) * | 1997-10-18 | 1999-04-29 | Boehringer Ingelheim International Gmbh | Tumour vaccine |
Non-Patent Citations (6)
| Title |
|---|
| Biodegradable micro- and nanoparticles as long-term deliveryvehicles for interferon-alpha. Alejandro Sanchez, Maria Tobio, Libia Gonzalez, et al.European Journal of Pharmaceutical Sciences,Vol.18 . 2003 |
| Biodegradable micro-and nanoparticles as long-term deliveryvehicles for interferon-alpha. Alejandro Sanchez, Maria Tobio, Libia Gonzalez, et al.European Journal of Pharmaceutical Sciences,Vol.18. 2003 * |
| Stabilization of recombinant interferon-α by pegylation forencapsulation in PLGA microspheres. Manish Diwan, Tae Gwan Park.International Journal of Pharmaceutics,Vol.252 . 2003 |
| Stabilization of recombinant interferon-αby pegylation for encapsulation in PLGA microspheres. Manish Diwan1&Tae Gwan Park.International Journal of Pharmaceutics,Vol.252 . 2003 |
| 重组人干扰素α长效注射微球的研究. 吴诚，7,18-20,33,63. 2004 |
| 重组人干扰素α长效注射微球的研究. 吴诚，7,18-20,33,63. 2004 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1754572A (en) | 2006-04-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| FI104954B (en) | Controlled drug delivery system comprising protein or polypeptide interaction and biodegradable polymers | |
| CA2338031C (en) | Biodegradable compositions for the controlled release of encapsulated substances | |
| Mwangi et al. | Synthesis and characterization of silk fibroin microparticles for intra-articular drug delivery | |
| Jameela et al. | Progesterone-loaded chitosan microspheres: a long acting biodegradable controlled delivery system | |
| Cui et al. | Preparation and characterization of melittin-loaded poly (DL-lactic acid) or poly (DL-lactic-co-glycolic acid) microspheres made by the double emulsion method | |
| ES2391315T5 (en) | Method for preparing microparticles having a selected polymer molecular weight | |
| Dong et al. | Stability of poly (D, L-lactide-co-glycolide) and leuprolide acetate in in-situ forming drug delivery systems | |
| ES2770273T3 (en) | Injectable administration of microparticles and compositions therefor | |
| BR122021025344B1 (en) | COMPOSITIONS COMPRISING SYNTHETIC NANOVEHICLES, THEIR USES, METHOD FOR PRODUCING THE SAME AND KIT | |
| US20090169632A1 (en) | Sustained release composition and manufacturing method thereof | |
| JP2001515862A (en) | Controlled release microsphere delivery system | |
| Yapar et al. | Injectable in situ forming microparticles: a novel drug delivery system | |
| PT3010962T (en) | Preparation of polylactide-polyglycolide microparticles having a sigmoidal release profile | |
| Küçüktürkmen et al. | In situ hydrogel formulation for intra-articular application of diclofenac sodium-loaded polymeric nanoparticles | |
| DK1532985T3 (en) | PROCEDURE FOR PREPARING A COMPOSITION WITH LONG-TERM RELEASE | |
| Alcalá-Alcalá et al. | A biodegradable polymeric system for peptide–protein delivery assembled with porous microspheres and nanoparticles, using an adsorption/infiltration process | |
| CN101797232B (en) | Method for preparing small-granularity recombination human vascular endothelium inhibin slowly-released particle for injection | |
| Chen et al. | Novel triptorelin acetate-loaded microspheres prepared by a liquid/oil/oil method with high encapsulation efficiency and low initial burst release | |
| Wichert et al. | Low molecular weight PLA: a suitable polymer for pulmonary administered microparticles? | |
| CN100475264C (en) | Slow release microphere for injection containing interferon or its analog, and preparation method thereof | |
| CN100528224C (en) | Slow release microphere for injection containing interferon alpha-1b and its preparation method | |
| Modi et al. | Copolymers of pharmaceutical grade lactic acid and sebacic acid: Drug release behavior and biocompatibility | |
| CN104013578A (en) | Paliperidone derivative slow release microsphere preparation and preparation method thereof | |
| Bhaskaran et al. | Biodegradable microspheres of ketorolac tromethamine for parenteral administration | |
| Shete | Microspheres as a unique drug carrier for controlled drug delivery: A review |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090819 Termination date: 20150927 |
|
| EXPY | Termination of patent right or utility model |