WO2007073035A1 - Procede de preparation de microspheres de polymere, et appareil de fabrication correspondant - Google Patents

Procede de preparation de microspheres de polymere, et appareil de fabrication correspondant Download PDF

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Publication number
WO2007073035A1
WO2007073035A1 PCT/KR2006/004263 KR2006004263W WO2007073035A1 WO 2007073035 A1 WO2007073035 A1 WO 2007073035A1 KR 2006004263 W KR2006004263 W KR 2006004263W WO 2007073035 A1 WO2007073035 A1 WO 2007073035A1
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WIPO (PCT)
Prior art keywords
polymer
peo
reaction chamber
solution
droplets
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PCT/KR2006/004263
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English (en)
Inventor
Sang-Jun Lee
Min-Hyo Seo
Bong-Oh Kim
Myung-Seob Shim
Hong-Ki Kim
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Samyang Corporation
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Priority to KR1020087014278A priority Critical patent/KR101140360B1/ko
Publication of WO2007073035A1 publication Critical patent/WO2007073035A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
    • A61K9/1647—Polyesters, e.g. poly(lactide-co-glycolide)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1682—Processes
    • A61K9/1694—Processes resulting in granules or microspheres of the matrix type containing more than 5% of excipient

Definitions

  • the present invention relates to a method and apparatus for preparing polymer microspheres using a biocompatible polymer. More particularly, the present invention relates to a method for preparing polymer microspheres, comprising the steps of injecting the biocompatible polymer solution through a nozzle with a predetermined pressure to form droplets; freezing the droplets by contacting the droplets with a liquefied gas to form frozen particles; removing the liquefied gas; and performing freeze-drying of the frozen particles, to prepare the polymer microsphere.
  • the present invention is characterized by preparing the polymer microspheres using a water-soluble biocompatible polymer under a low temperature, without using any organic solvent.
  • microspheres As a drug delivery system.
  • the microspheres have been applied to a drug which may exhibit its therapeutic effect only when administrated repeatedly, and was applied first to a drug for prostate cancer.
  • LHRH which is one of sex hormones needs to be administrated periodically in order to suppress a growing of the prostate cancer.
  • a frequent administration of the LHRH is very inconvenient to patients, and thus a microsphere formulation that may be effective for a month or more by one administration was developed.
  • Y. Ogawa, et al. introduced a preparing method for a leuprorelin acetate depot formulation using a polymer in Chem. Pharm. Bull, 36, 2576-2581(1988). That is, the preparation was performed by dissolving the leuprorelin acetate in an aqueous gelatin solution, and mixing the solution with a methylene chloride solution containing a polylacticglycolic acid (PLGA) to form a W/O emulsion. Sequently, the mixed emulsion was dropped into 0.25% aqueous PVA solution to form a W/O/W emulsion. Then, methylene chloride was removed and freeze-drying was performed to remove remained water.
  • PLGA polylacticglycolic acid
  • the microsphere prepared by the method of Y. Ogawa, et al. continuously released the leuprorelin acetate over a month with one injection.
  • the LHRH is a peptide drug with a molecular weight of 1100, and thus, has no difficulty in being formed as microspheres.
  • protein drugs with a molecular weight of 1100 or more may have a high chance to lose their specific activities by reacting with an organic solvent or a temperature variation while being produced as microspheres.
  • US Patent No. 5,922,253 discloses a method and apparatus related to a cryogenic process which is one of processes for preparing the hGH microspheres.
  • a PLGA or a polylactic acid (PLA) is dissolved in an organic solvent, and the solution is sprayed over a region that maintains a low temperature using a liquefied gas, thereby being frozen. Then, the frozen droplets of the solution are precipitated in a solvent which may not dissolve the PLGA or the PLA in order to extract the initial organic solvent.
  • These processes have many advantages, such as a high yield and formation of microparticles that may be used commercially.
  • these processes may be performed under a sterile state, and a size of the microparticle may be controlled.
  • these processes also use the organic solvent, thereby having a chance that a protein is denaturalized.
  • An activity of a peptide or a protein drug may be reduced due to several factors, and particularly, such activity reduction becomes great when being contacted with an organic solvent, subjected to pH variation, or exposed to a shear force.
  • a thermosensitive polymer hydrogel has been developed.
  • a polymer used in this formulation exhibits a sol-gel phase transition depending on temperature when dissolved in water.
  • This polymer is a liquid phase at a low temperature, whereby it is capable of being mixed well with a protein or a peptide drug, and becomes a solid phase by a body temperature when administrated into a body, thereby slowly releasing the protein or the peptide.
  • any organic solvents that reduce an activity of the protein are not used, thereby minimizing the reduction of the activity.
  • An exemplary polymer developed for this hydrogel system is poloxamer 407.
  • the poloxamer 407 is a block copolymer of polyethylene glycol (PEG) and polypropylene glycol (PPG), and has a liquid phase when being dissolved in water at a low temperature and phase-changed to a solid gel phase when the temperature increases.
  • PEG polyethylene glycol
  • PPG polypropylene glycol
  • Such polymer is called as a thermosensitive polymer, and many studies for using the polymer solution as a peptide or a protein delivery system have been continued.
  • the concentration of the polymer solution is
  • US Patent No. 6,004,573 discloses a block copolymer consisting of PEG and PLGA.
  • the copolymer has PEG-PLGA-PEG composition and exhibits a phase transition similar to that of the poloxamer 407. That is, the copolymer is in a liquid sol phase at a low temperature and phase-changed into a solid gel phase when temperature increases.
  • the PEG-PLGA-PEG copolymer was studied as a formulation of releasing a paclitaxel, which is a water-insoluble anticancer agent, for one month.
  • An aqueous solution of the PEG-PLGA-PEG copolymer exhibiting the sol-gel phase transition may be used as a delivery system for a protein drug, due to the properties that the aqueous solution of the copolymer is in a liquid sol phase at a low temperature, thereby being easily mixed with a protein, and phases-changed into a solid phase by a body temperature when administrated into a body, thereby releasing the protein slowly.
  • a polymer composition consisting of the poloxamer 407 that is biodegradable in the body is disclosed in Controlled release society 3(? h annual meeting Proceedings # 167 by X. Zaho et al. According to X. Zaho et al., the poloxamer 407 is reacted with a disuccinimidyl carbonate (DSC) to prepare a compound wherein the poloxamer 407s are connected by degradable carbonate bonds.
  • DSC disuccinimidyl carbonate
  • US Patent No. 6,348,558 discloses a degradable polymer consisting of at least two polyalkylene oxide oligomers connected by hydrolysable carbonate bonds.
  • the object of the present invention is to provide a method for preparing microspheres using a polymer solution without using any organic solvents.
  • Another object of the present invention is to provide an apparatus for preparing microspheres using a polymer solution without using any organic solvents.
  • FIG. 1 is a schematic view of an apparatus for preparing microspheres by a direct injection method.
  • FIG. 2 is a schematic view of an apparatus for preparing microspheres by an indirect injection method.
  • FIG. 3 is a SEM image of a prepared microsphere.
  • FIG. 4 is a IH-NMR spectrum of a poloxamer disuccinate.
  • reaction chamber 20 reaction chamber 20: vacuum pump 22: vacuum tube
  • injection unit 40 injection unit 40: nozzle 42: pressure control pump
  • chamber side heat exchanger 62 nozzle side heat exchanger
  • Gl polymer solution
  • G2 spheres (frozen particles) DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
  • the present invention relates to a method of preparing a microsphere comprising the steps of: injecting a biocompatible polymer solution through a nozzle with a predetermined pressure to form droplets; freezing the droplets by contacting the droplets with a liquefied gas to form frozen particles; removing the liquefied gas; and performing freeze-drying of the frozen particles, to prepare the polymer microsphere, and an apparatus for preparing the polymer microsphere.
  • the microsphere is prepared by freezing and freeze-drying the droplets formed by injecting a biocompatible polymer solution without using an organic solvent.
  • the biocompatible polymer is water-soluble, preferably has a high molecular weight, and more preferably exhibits a sol-gel phase transition which is a sol phase at a low temperature and becomes a gel phase at a high temperature.
  • the biocompatible polymer is a multi-block copolymer as mentioned below. The following will describe definition of terminologies used in the present invention.
  • biodegradable or biocompatible means a property to be hydrolyzed or degraded by enzymes, thereby being absorbed into the body or safely eliminated from the body, after administrated into the body.
  • microsphere means a fine sphere having the size of lmm or less.
  • the microsphere is a sphere-shaped fine particle, generally called as microparticle, which has the size ranging from several ⁇ m to several hundreds ⁇ m.
  • polyxamer is a compound wherein hydrophilic polyethylene oxide (PEO) blocks and hydrophobic polypropylene oxide (PPO) blocks are connected as the form of a PEO-PPO-PEO tri-block copolymer by ether bonds.
  • the poloxamer has the weight average molecular weight of 1,000 to 20,000 Daltons and has hydroxyl groups at both terminuses.
  • a poloxamer 188 (Pluronic® F-68) and a poloxamer 407 (Pluronic® F- 127) which are commercialized compounds may be used.
  • sol-gel phase transition means a reversible reaction that a sol phase is maintained at a specific temperature or below and becomes changed into a gel phase at specific temperature or above, or a gel phase is maintained at a specific temperature or above and becomes changed into a sol phase at the specific temperature or below.
  • specific temperature may be varied depending on the types of polymers used, the concentration of an aqueous solution, the presence or absence of a salt, and the concentration of hydrogen ion, a range of the specific temperature may be 5 to 37 C, and preferably, 25 to 35 ° C.
  • freeze-drying generally means a drying method by dissolving various materials in water, freezing the solution at 0 ° C or below, and reducing the pressure using a vacuum pump, thereby generating sublimation of the water used as a solvent.
  • the present invention provides a method of preparing microspheres, comprising the steps of: injecting a biocompatible polymer solution through a nozzle with a predetermined pressure to form droplets; freezing the droplets by contacting the droplets with a liquefied gas to form frozen particles; removing the liquefied gas; and performing freeze-drying of the frozen particles, to prepare the polymer microsphere.
  • the method of preparing the microspheres according to the present invention comprises the steps of: i) injecting a biocompatible water-soluble polymer solution through a nozzle with a predetermined pressure to form droplets; ii) freezing the droplets by contacting the droplets with a liquefied gas to form frozen particles; iii) removing the liquefied gas and performing freeze-drying of the frozen particles to prepare the polymer microsphere.
  • the method of preparing the microspheres according to the present invention comprises the steps of: i) injecting a biocompatible water-soluble polymer solution having a high molecular weight through a nozzle with a pressure ranging from 0.5 to 15bar to form droplets; ii) freezing the droplets by contacting the droplets with a liquefied gas to form frozen particles; and iii) removing the liquefied gas and performing freeze-drying of the frozen droplets at a temperature ranging from -70 to -1 C and a pressure of lTorr or less.
  • the droplets may be formed by injecting a polymer solution through a nozzle with a predetermined pressure at a specific temperature or below and contacting the formed droplets with a liquefied gas, thereby forming frozen particles having the size ranging from several ⁇ m to several hundreds ⁇ m. Thereafter, the liquefied gas is removed, and a freeze-drying of the frozen particles is performed by reducing the pressure at a low temperature, thereby preparing the polymer microspheres.
  • the present invention provides an apparatus for preparing polymer microspheres, which comprises a reaction chamber, an injection unit, and a temperature control unit.
  • the apparatus according to the present invention is for preparing polymer microspheres from a biocompatible polymer solution, and comprises: a reaction chamber connected with a vacuum pump through a vacuum tube provided with an exhaust hole through which inner gas can be exhausted and an inclined bottom surface inclined toward a discharge hole; an injection unit including the nozzle penetrating and extending into the reaction chamber, the nozzle being connected with a pressure control pump to inject the polymer solution supplied under a predetermined pressure into the reaction chamber, thereby forming a uniform droplet; and a temperature control unit having a chamber side heat exchanger installed at a space formed on a wall of the reaction chamber and a nozzle side heat exchanger surrounding an exposed region of the nozzle out of the reaction chamber, these heat exchanger circulating a heat exchange medium pressure-fed by the temperature control unit, thereby maintaining a temperature around the reaction chamber and the nozzle below a predetermined temperature and preventing the polymer solution from being phase-changed into a gel state.
  • FIGs. 1 and 2 An apparatus for preparing microspheres according to an embodiment of the present invention is shown in FIGs. 1 and 2. An apparatus and method for preparing polymer microspheres will now be described with reference to FIGs. 1 and 2.
  • an apparatus for preparing a polymer microsphere comprises a reaction chamber 2, an injection unit 4 disposed at an outer side of the reaction chamber 2 and injecting a polymer solution into the reaction chamber 2, and a temperature control unit 6 maintaining an inside temperature of the reaction chamber 2 within a predetermined range.
  • the injection unit 4 disposed at a top-center or side of the reaction chamber 2 includes a nozzle 40 extending into the reaction chamber, and a pressure control pump 42 providing the polymer solution injected through the nozzle and forming uniform droplets in the reaction chamber 2.
  • the reaction chamber 2 may be a 20-30L cylinder, and formed of a material enduring a vacuum state below ltorr, such as steel or stainless steel.
  • a diameter of the nozzle 40 affects a size of the droplet, and thus may be varied according to a desired droplet size.
  • the diameter of the nozzle 40 may be ranging from 0.1mm to lmm.
  • a heat exchange medium may be disposed in a space defined between inner and outre surfaces of the reaction chamber 2.
  • a nozzle or an atomizer which is usually used in a spray-dryer may be used for the nozzle 40.
  • the atomizer may be an external air atomizer or an internal air atomizer, and the nozzle may be a spray-dryer such as a bucci.
  • the temperature control unit 6 includes a jacket or pipe shaped chamber side heat exchanger 60 installed at a space formed in a wall of the reaction chamber 2, and a pipe shaped nozzle side heat exchanger 62 surrounding the exposed nozzle 40 out of the reaction chamber 2.
  • a heat exchange medium is provided and circulated to these heat exchangers 60 and 62 by a temperature control unit 64 installed separately, thereby removing a liquefied gas, maintaining a temperature around the reaction chamber 2 and the nozzle 40 at a predetermined level of -40— 5 ° C, and preventing the polymer solution or the droplet from being phase-changed into a gel state.
  • a typical refrigerating unit may be used as the temperature control unit 64, and, in this case, a CFC-based refrigerant such as HCFC22, HFC 134a, R410a, or R407c may be used.
  • a CFC-based refrigerant such as HCFC22, HFC 134a, R410a, or R407c may be used.
  • a stirring blade 32 may be driven if frozen spheres that are frozen particles, are aggregated at a bottom of the reaction chamber 2 during the freezing of the droplets of the polymer solution by contacting the liquefied gas.
  • a motor or a decelerator (not shown) may be used to rotate the stirring blade 32.
  • the stirring blade 32 may be a propeller type, a vane type, a turbine type, a spiral type, an oar type, or an impeller type, and an embodiment of the present invention may use the propeller type stirring blade.
  • an upper side of the reaction chamber 2 may have a lid-structure which may be opened or closed. This may require a gasket or a packing at the lid in order to maintain a sealed state, which is for preventing an external air from inflowing inside, thereby enhancing a vacuum state of an inner space of the reaction chamber 2.
  • a handle 34 may be attached on a proper place of the reaction chamber 2 in order to have easy dissembling or assembling of the lid.
  • An exhaust hole 24 may be disposed at the top of the reaction chamber 2 for a vaporized gas generated from the liquefied gas. After the droplets of the injected polymer solution are frozen to form frozen particles by the liquefied gas, the vaporized gas from the liquefied gas is exhausted through the exhaust hole 24 during the removing of the liquefied gas. Further, when the droplets of the polymer solution contact the liquefied gas disposed at the bottom of the reaction chamber 2, a gas generated from the liquefied gas is exhausted through the exhaust hole 24, thereby preventing an increase of an inner pressure of the reaction chamber 2 caused by a large volume of the gas generated from the liquefied gas contacting a large volume of the droplets.
  • a bottom of a storage space storing the heat exchange medium may be inclined in order to collect the formed microspheres easily.
  • the preparing of the microspheres starts in a state where the inner space of the reaction chamber 2 is kept to a degree of vacuum below ltorr by a vacuuming operation of a vacuum pump 20 through a vacuum tube 22 and the liquefied gas is filled at the bottom of the reaction chamber 2 with a predetermined level.
  • a method for preparing the microspheres of the present invention comprises a step for forming the droplets by injecting the polymer solution through the nozzle 40 with the predetermined pressure.
  • An injection method of the polymer solution may be classified into a direct injection and an indirect injection depending on a position of an injection nozzle.
  • the direct injection has the nozzle 40 disposed at the top of the reaction chamber 2. Therefore, when the polymer solution Gl is injected, the droplets travel in the same direction as gravity, thereby rapidly contacting the liquefied gas disposed at the bottom and forming frozen spheres G2, that are frozen particles.
  • the direct injection may quickly freeze a large volume of the aqueous polymer solution, but an aggregation of frozen droplets may happen unless agitated enough.
  • the indirect injection has the nozzle 40 disposed at the side of the reaction chamber 2.
  • the indirect injection injects the polymer solution under a lower pressure than the pressure of the direct injection. Therefore, the injected polymer solution Gl travels in the horizontal direction to the nozzle 40 and in the perpendicular direction to the gravity for a while, then travels to the bottom of the reaction chamber 2 by the gravity thereby contacting the liquefied gas disposed at the bottom and forming frozen spheres G2 that are frozen particles.
  • the indirect injection has a slower freezing process, producing a less amount of the aggregated frozen droplets, but it needs a long time to freeze the droplet.
  • All synthetic or natural water-soluble polymers may be used as a water-soluble polymer of the present invention.
  • the polymers may have a high molecular weight and more preferably, exhibit a sol-gel phase transition.
  • the exemplary example of such polymers may be a multi-block copolymer described hereinafter.
  • the polymer of the present invention may be selected from the group consisting of hyaluronic acid, dextran, gelatin, collagen, chitosan, methylcellulose (MC), ethylcellulose (EC), hydroxyethylcellulose (HEC), methylhydroxyethylcellulose (MHEC), hydroxymethylcellulose, hyroxypropylmethylcellulose, hyroxypropylcellulose, and a multi-block copolymer wherein tri-block copolymers are connected through a dicarboxylic linker, and each tri-block copolymer has two polyethylene oxide (PEO) blocks and a polypropylene oxide (PPO) or a polybutylene oxide (PBO) that is positioned between the two PEO blocks.
  • PEO polyethylene oxide
  • PPO polypropylene oxide
  • PBO polybutylene oxide
  • microspheres having more densified structure may be obtained.
  • the weight average molecular weight of the polymer may be from 25,000 to 1,000,000 Daltons, and preferably from 40,000 to 1,000,000 Daltons.
  • the microspheres, which are prepared from a polymer with a low molecular weight, such as poloxamer have a problem to be dissolved before being injected into the body after reconstituted in an aqueous solution.
  • microspheres prepared from a polymer with a high molecular weight have a high strength, and thus, is capable of being injected into the body with maintaining the spherical shape.
  • the polymer solution may be obtained by dissolving the water-soluble polymer into an aqueous solvent, such as distilled water, an acetate buffer solution, a phosphate buffer solution, or a peptide or a protein solution.
  • a concentration of the polymer solution may be from 3% to 25% (w/w), preferably from 3% to 15% (w/w). When the concentration of the polymer solution is lower than the range, a particle may not be formed, and when the concentration is higher than the range, an excessively high pressure is required to spray the polymer solution.
  • the water-soluble polymer solution used in the present invention may exhibit the sol-gel phase transition, which may be applied as a good drug delivery material capable of a controlled release of a drug when administrated into the body with the drug included therein.
  • the water-soluble polymer exhibiting the sol-gel phase transition may be preferably a multi-block copolymer as described hereinafter.
  • the water-soluble polymer solution preferably the multi-block copolymer solution becomes a gel phase at a higher temperature than a specific sol-gel phase transition temperature due to its sol-gel phase transition property. Therefore, the injection temperature of the multi-block copolymer solution may be maintained below the sol-gel phase transition temperature. When the injection temperature is maintained below the sol-gel phase transition temperature, a viscosity of the multi-block copolymer solution is lowered, whereby the microsphere may be formed. According to the present invention, the injection temperature of the multi-block copolymer solution may be maintained 15 ° C or below, preferably from 5 ° C to 12 ° C. Because the polymer solution Gl exhibits the sol-gel phase transition, the temperature of the solution may also be maintained below a specific temperature by the nozzle side heat exchanger 62 that is installed around the nozzle 40, thereby preventing gelation caused by increased temperature during the injection.
  • the polymer solution is injected through the nozzle 40 under a predetermined pressure provided from a pressure control pump 42, and a size of the droplet formed by the injection is reduced when an injection pressure increases. Therefore, the size of the droplet may be controlled by varying the injection pressure. Further, the injection speed may also be controlled by varying the injection pressure.
  • the injection pressure may be determined considering the size of the droplet to be prepared, the injection speed, and the like. For example, the injection pressure may be from 0.5bar to 15bar, and preferably from lbar to 5bar. If the injection pressure is lower than the range, an injection of the aqueous solution may not occur, or the size of the droplet increases and a long time is required for the injection. If the injection pressure is higher than the range, the formed droplets may be aggregated since the injection speed is too high.
  • the droplets formed by injecting the polymer solution Gl fall to the bottom of the reaction chamber 2, and are changed into spheres G2, i.e., frozen particles by being contacted with the liquefied gas.
  • the droplets of the polymer solution injected from the nozzle 40 are rapidly frozen as soon as being contacted with the liquefied gas, which forms the frozen particles with a same size of the droplet formed by the injection. Therefore, by controlling the size of the droplet formed by the injection, the size of the frozen particle may be controlled, and the size of the droplet is controlled by varying a diameter of the injection nozzle and/or the injection pressure. It is preferable to stir the liquefied gas in order to prevent the aggregation of the droplets during the freezing of the droplets by contacting with the liquefied gas. If the stirring speed is too high, the frozen particles G2 of the droplets may be aggregated.
  • the stirring speed may be determined considering the kind of the polymer solution to be injected and an injection method.
  • the preferable stirring may be from 5rpm to 300rpm, and this may be varied according to a viscosity or a concentration of the polymer solution to be injected, or manufacturing processes. More preferably, the stirring speed may be from 50rpm to lOOrpm during the injection of the polymer solution, and from 60rpm to 150rpm after completion of the injecting.
  • the liquefied gas is not limited, and preferably, has a boiling point of -20 C or below in order to freeze the droplets rapidly. More preferably, the liquefied gas may be selected from the group consisting of liquefied nitrogen, liquefied oxygen, and liquefied helium.
  • the method of preparing the microspheres according to the present invention includes a freeze-drying process after removing the liquefied gas from the frozen droplets, i.e., the frozen particles G2. More specifically, the frozen spheres, i.e., the frozen particles G2, are freeze-dried in a freeze-drying condition formed by the chamber side heat exchanger 60, the temperature control unit 64, and the vacuum pump 20, thereby producing porous microspheres. The produced microspheres are collected through an outlet 26 after rolling along the inclined surface 28 of the bottom of the reaction chamber 2.
  • the temperature control unit 64 connected with the chamber side heat exchanger 60 located at the bottom of the reaction chamber 2 is driven, to increase a temperature of the bottom of the reaction chamber 2 to a level higher than the boiling point of the liquefied gas, whereby the liquefied gas is vaporized and removed.
  • the vaporized gas is discharged through the exhaust hole 24 located at the top of the reaction chamber 2.
  • the temperature of the bottom of the reaction chamber 2 is maintained higher than the boiling point of the liquefied gas.
  • the temperature of the bottom of the reaction chamber 2 may be from -70 ° C to -1 ° C, preferably from -40 ° Cto -5 ° C, further preferably about -20 " C.
  • the temperature of the bottom of the reaction chamber 2 from -70 C to -1 C, preferably from -40 C to -5 C, and further preferably about -20 C, and to reduce the pressure of the inner space of the reaction chamber 2 to ltorr or below by driving the vacuum pump 20.
  • the temperature may be about -70 to-1 ° C, preferably -40 to -5 ° C, and the pressure may be ltorr or less. If the temperature and the pressure of the inner space of the reaction chamber 2 are higher than the range above, the frozen particles may be aggregated or melt, which may not allow maintaining the spherical shape.
  • the freeze-drying may be performed using the vacuum pump 20 for 6 to 12 hours, and the temperature for the initial 3hrs may be -70 to -1 C, preferably -40 to -5 C, more preferably about -20 C. Thereafter, the freeze-drying may be further performed with increasing the temperature slowly up to a room temperature.
  • a size of the produced microsphere may be controlled by the diameter of the nozzle 40 and the pressure generated during the injection process.
  • the size of the produced microsphere may be 10 to 500 ⁇ m, and provided with a porous surface.
  • the porous surface is formed by the sublimation of water contained in the polymer solution during the freeze-drying.
  • a biocompatible polymer solution may include further bioactive materials.
  • Drugs useful in the present invention may include protein or peptide drugs selected from the group consisting of various protein or peptide hormones, various antibiotics, growth hormone (GH), interferon (INF), Granulocyte colony stimulating factor (G-CSF), erythropoietin (EPO), interleukin (IL), follicle stimulating hormone (FSH), nerve growth factor (NGP), octreotide, insulin, calcitonin, tumor necrosis factor (TNF), vascular endothelial growth factor (VEGF), epithelial growth factor (EGF), platelet derived growth factor (PDGF), bone morphogenic protein (BMP), tissue plasminogen activator (TPA), and oligonucleotide.
  • the protein or the peptide may be contained in the buffer solution.
  • the biocompatible polymer When the biocompatible polymer may be dissolved into the buffer solution including the protein or the peptide drug(s) and then, freeze-dried, the accordingly obtained microsphere containing the protein or the peptide drug(s) may be stored for a long period since there is no water therein.
  • the content of the protein or the peptide drug may be 0.01 to 50 % by weight based on the weight of the polymer.
  • the content of the protein or peptide drug may be controlled considering the releasing time, the releasing type, and the like. It is preferable that the content thereof may be 0.1 to 20 % by weight in order for the prepared microspheres to properly maintain their manufacturing features or shape.
  • microspheres of the present invention may be used in the form of microspheres themselves or nanospheres. Further, the microspheres of the present invention may be applied as a controlled-released drug delivery system formulated in the form of a hybrid sol-gel depot system for drug delivery, a strip type, a bar type or a film type.
  • the microspheres of the present invention may further include the polymer selected from the group consisting of PEG, hyaluronic acid, dextran, gelatin, collagen, chitosan, poloxamer 407, poloxamer 188, methylcellulose, ethylcellulose (EC), hydroxyethylcellulose (HEC), methylhydroxyethylcellulose (MHEC), hydroxymethylcellulose, hydroxypropylmethylcellulose (HPMC), and hydroxypropylcellulose.
  • the polymer may be additionally contained in the polymer solution in the amount of 0.1 to 50% by weight of based on the weight of the polymer.
  • the microspheres of the present invention may be applied as a hybrid thermosensitive depot system, a microsphere type, a nanosphere type, a strip type, a bar type, or a film type.
  • the gelation temperature or the gel strength of the polymer may be varied when preparing the hybrid-type drug delivery system.
  • various excipients and/or stabilizers for stabilizing the protein or the peptide drugs may be further added.
  • the stabilizers and the excipients allow the protein or the peptide drugs to retain their activity without denaturing in the polymer solution, and supply a buffering effect during the freeze-drying process for preparing the microspheres, thereby maintaining a specific activity of the protein or the peptide drug.
  • the stabilizer may be selected from the group consisting of a phosphate buffer solution (PBS), amino acids, carbohydrates, fatty acids, and surfactants.
  • the excipient may be selected from the group consisting of lactose, dextran, manitol, and sucrose.
  • amount of the stabilizer and/or the excipient may be varied according to a property of a desired microsphere. Since the physical property of the microsphere may be changed by an excessive amount of the stabilizer and/or the excipient, it is required to maintain the amount of the stabilizer and/or the excipient within a specific range, preferably 1 to 20% by weight based on the weight of the polymer.
  • the proteins or peptides When a metal cation is added into some proteins or peptides, the proteins or peptides may exhibit a high stability even after the freeze-drying, and control the release rate.
  • the addition amount of the cation is varied depending on the type of the proteins or the peptides.
  • the proteins or the peptides and the metal cation may be mixed at the mole ratio of about 1:10 to 1:5000 (mole of the protein or the peptide : mole of the metal cation).
  • the metal cation having the effect described above may be Ca 2+ , Zn 2+ , or Mg 2+ .
  • An administration method and dosage of the drug delivery system is determined according to a medicinal activity, a target site in the body, physiochemical characteristics, and the like.
  • the administration method may be an oral, a subcutaneous, an intravenous, or an intraperitoneal administration.
  • a conventional method for preparing a microsphere includes dissolving a PLGA or PLA into an organic solvent, dropping the solution into an aqueous solution, forming spherical droplets, and removing the organic solvent.
  • a preparing method of the present invention includes dissolving a biocompatible polymer such as a multi-block copolymer into an aqueous solvent instead of the organic solvent, forming droplets by injecting the solution; and performing a freeze-drying using a liquefied gas.
  • the microspheres produced by the method of the present invention is not harmful to a human body and prevents the denaturation or deterioration of an activity of a bioactive drug such as a protein which may be included in the microspheres, thereby enhancing a stability of the drug.
  • the controlled-release depot-type formulation may be prepared by mixing a bioactive drug such as a protein or a peptide after dissolving the biocompatible polymer of the present invention in the aqueous solvent such as water.
  • the sol phase- polymer solution may be administrated by the subcutaneous injection with a syringe, therefore a temperature of the syringe storing the polymer solution needs to be maintained at a low temperature until the polymer solution is administrated.
  • the sol phase polymer solution stored in the syringe may be phase-changed into a solid phase, thereby making it difficult to perform the administration thereof.
  • the sol-gel phase transition may be avoided until the administration when the formulation is produced in the form of the microspheres.
  • the depot formulation When the depot formulation is produced using the multi-block copolymer of the present invention, water may be included in the depot formulation, whereby hydrolysis may occur, and thus, it is difficult to be stored for a long period. Also, in the case of a freeze-storage, if the formulation is not formed in the microsphere type, the formulation becomes hardened, thereby having a difficulty in thawing. However, the microsphere formulation of the present invention does not include the water and is formed in a relatively small size, thereby capable of solving the problems. Further preferably, the biocompatible polymer of the present invention may be a multi-block copolymer as described below.
  • the multi-block copolymer is an ionic polymer which is in a sol state at a low temperature and phase-changed into a gel state at a high temperature, wherein tri-block copolymers are connected through dicarboxylic linkers, and each tri-block copolymer has two PEO blocks and a PPO block or a PBO block that is positioned between the two
  • the multi-block copolymer of the present invention forms hydrogel at a specific concentration and a specific temperature, exhibits a sol-gel phase transition, and is biodegradable. Also, the multi-block copolymer is formed by coupling through the dicarboxylic linkers, thereby increasing the molecular weight and thus, enhancing a durability of the gel. Further, since the multi-block copolymer has an ionic terminal group, the drug can be slowly released from the gel.
  • the multi-block copolymer may be represented by the following Chemical Formula 1.
  • PEO is a polyethylene oxide block
  • Y is PPO(polypropylene oxide block), PBO(polybutylene oxide block), or a combination thereof
  • X is H or an anionic group
  • n is an integer between 1 and 100, preferably between 3 and 100;
  • R is-(CH 2 ) m - or an aryl group of -C m '-, wherein m is an integer between 0 and 20 and m' is an integer between 6 and 12;
  • M is H or a cationic group preferably selected from the group consisting of Li, Na, K, Ag, Au, Ca, Mg, Zn, Fe, Cu, Co, and Ni, when X is not H; and
  • M does not be present when X is H.
  • the polymer of the present invention may be represented as the following Chemical Formula 2.
  • Y is PPO, PBO, or a combination thereof
  • n is an integer between 1 and 100, preferably between 3 and 100;
  • R is -(CH 2 ) m - or an aryl group of -C m -, wherein m is an integer between 0 and 20 and m' is an integer between 6 and 2; if X is not H, M is H or a cationic group having a univalent or divalent preferably selected from the group consisting of Li, Na, K, Ag, Au, Ca, Mg, Zn, Fe, Cu, Co, and Ni, if X is H, M does not exist.
  • the PEO block of the polymer described above may consist of ethylene oxide units having a unit number ranging from 2 to 2000, preferably from 5 to 500, and more preferably from 80 to 120.
  • the ethylene oxide unit number of each of the two PEO blocks may be same as or different from each other.
  • the PPO or the PBO block may have 2 to 200 units, preferably 20 to 500 units, further preferably 30 to 250 units.
  • the multi-block copolymer includes at least 2 PEO-PPO (or PBO)-PEO units, and each unit may be such as PEO-PPO-PEO, PEO- PBO-PEO, or a PEO-(a combination of PPO and PBO)-PEO.
  • the multi-block copolymer of the present invention may have a weight average molecular weight of 25,000 to 1,000,000 Daltons, preferably 40,000 to 1,000,000 Daltons, more preferably 40,000 to 500,000 Daltons, and further preferably 80,000 to 120,000 Daltons.
  • multi-block copolymer in the present invention refers to a copolymer wherein a polyethyleneoxide block is linked to a polypropyleneoxide or polybutyleneoxide block, which is, in turn, linked to a polyethyleneoxide block and the resulting at least 2 PEO-PPO (or PBO)-PEO blocks are connected through biodegradable dicarboxylic linkers.
  • a ratio of the molecular weight between PEO and PPO or PBO may be varied so long as the water-soluble property of the polymer is retained, for example ranging from 0.2:1 to 40:1, preferably 1:1 to 7.5:1, and more preferably 1:1 to 5:1.
  • the PEO block may be contained in the amount of 10 to 85% by weight, preferably 40 to 85% by weight, based on a PEO-PPO (or PBO)-PEO unit.
  • the multi-block copolymer of the present invention i.e., the PEO-PPO (or PBO)-PEO unit is connected through the dicarboxylic linker capable of being hydrolyzed in the body.
  • dicarboxylic linker used in the present invention refers to an alkyl or an aryl compound having 2 carboxyl groups in a molecule, such as oxalic acid, malonic acid, succinic acid, adipic acid, etc, and preferably, a nontoxic compound to the human body.
  • the dicarboxylic linker may be selected from the group consisting of alkyl dicarboxylic acids including malic acid, oxalic acid, succinic acid, glutaric acid, adipic acid, sebacoyl acid, suberic acid, dodecanoic acid, and the like; unsaturated dicarboxylic acids including fumaric acid, maleic acid, and the like; and aryl dicarboxylinc acids including phthalic acid, terephthalic acid, and the like.
  • alkyl dicarboxylic acids including malic acid, oxalic acid, succinic acid, glutaric acid, adipic acid, sebacoyl acid, suberic acid, dodecanoic acid, and the like
  • unsaturated dicarboxylic acids including fumaric acid, maleic acid, and the like
  • aryl dicarboxylinc acids including phthalic acid, terephthalic acid, and the like.
  • the dicarboxylic linker may be connected with hydroxyl groups positioned at both termini of PEO-PPO (or PBO)-PEO by ester bonds, and the ester bonds may be hydrolyzed in an aqueous solution or in the body or degraded by an enzyme, whereby the multi-block copolymer is degraded to carboxylic acid and PEO-PPO (or PBO)-PEO unit.
  • the both terminal ends of the multi-block copolymer of the present invention are hydroxyl or ionic groups.
  • a salt corresponding to the anionic group may be a monovalent metal cation such as Li, Na, K, Ag, or Au, or a divalent cation such as Ca, Mg, Zn, Fe, Cu, Co, or Ni.
  • the polymer having anionic groups at the both terminal ends forms a complex by reacting with divalent cation
  • the polymer may maintain the more stable gel state, thereby capable of continuously releasing a drug therefrom.
  • an ion salt may be formed, thereby decreasing the initial burst rate of the drug from the multi-block copolymer gel, and enhancing the durability of the drug release.
  • the multi-block copolymer of the present invention can be effectively applied as a non-ionic and ionic drug delivery system.
  • the multi-block copolymer PEO- PPO (or PBO)-PEO may be a commercialized poloxamer.
  • the poloxamer is a tri-block copolymer having a hydrophilic PEO block and a hydrophobic PPO block that are interconnected in the PEO-PPO-PEO tri-block form by ether bonds.
  • the poloxamer has a weight average molecular weight of 1,000 to 20,000 Daltons and has hydroxyl groups as both terminal ends.
  • the poloxamer may be poloxamer 188 (Pluronic ® F-68), or poloxamer 407 (Pluronic® F-127).
  • the poloxamer may or may not be purified, but a polymer having a high molecular weight may be produced easily when the poloxamer is purified.
  • the purification of the poloxamer may be implemented by precipitating in hexane after dissolving the poloxamer in methylene chloride, or by a phase separation in n- propanol/H 2 O solvent as disclosed in US Patent No. 5,800,711.
  • the method of preparing a multi-block copolymer with terminal hydroxyl ends comprising:
  • the method of preparing a multi-block copolymer with terminal carboxyl ends comprising:
  • the polymer of the present invention having carboxyl groups at both terminal ends may be dissolved in a solvent capable of being mixed with water, such as acetone, acetonitrile, or dioxane, and subsequently, neutralized with sodium carbonate or sodium hydrogen carbonate, thereby preparing a multi-block poloxamer having a sodium dicarboxylic acid salt at both terminal ends.
  • a solvent capable of being mixed with water such as acetone, acetonitrile, or dioxane
  • the polymer having the sodium dicarboxylic acid salt at the both terminal ends may be treated with an aqueous solution of calcium chloride, zinc chloride, magnesium chloride, iron chloride, copper chloride, silver nitrate, potassium chloride, or lithium chloride, and then, a dialysis be preformed, to produce a polymer having metal carboxylate groups at both terminal ends.
  • a multi-block copolymer having a sulfate or phosphate groups at terminal ends may be produced by a method including:
  • a multi-block copolymer having other anionic groups at terminal ends may also be produced by any method well-known in the relevant art using the multi-block copolymer of the present invention.
  • a dicarboxylic dihalide may be directly reacted as a dicarboxylic linker, and if a dicarboxylic acid is a starting material, the dicarboxylic acid may be activated by oxalic chloride which transforms the dicarboxylic acid to a dicarboxylic dichloride.
  • the reaction may be performed with a solvent or without any solvent, and preferably, the solvent may be selected from the group consisting of dichloromethane, chloroform, tetrahydrofuran, acetonitrile, acetone, toluene, and dioxane.
  • the solvent may be selected from the group consisting of dichloromethane, chloroform, tetrahydrofuran, acetonitrile, acetone, toluene, and dioxane.
  • the polymerization rate and degree capable of determining the average molecular weight of the polymer of the present invention may be controlled by controlling the reaction temperature and time.
  • the reaction temperature may be varied depending on the boiling point of the solvent used, preferably the reaction temperature may be within the range of 60 to 12O 0 C, and the reaction time may be within the range of 12 to 72 hours.
  • a catalyst such as tin octoate, zinc chloride, and the like, may be used in order to increase the reaction rate.
  • the reaction rate may also be increased by using an amine such as pyridine, dimethylaminopyridine, imidazole, triethylamine, and the like, in the amount of 2 equivalents based on 1 equivalent of the dicarboxylic acid.
  • an amine such as pyridine, dimethylaminopyridine, imidazole, triethylamine, and the like
  • the polymer may be purified by any known method in the relevant art, and preferably by a precipitation using a solvent that dissolves the reactant but does not dissolve the product.
  • the polymer of Chemical Formula 1 dissolved in distilled water with the concentration of 2 to 40% forms a sol state at a temperature of 2 to 8 0 C and phase- changed to a gel state at a temperature of 30 to 50 0 C.
  • a polymer of the present invention may form a gel state at a lower concentration of 10% compared to poloxamer, thereby reducing toxicity to the body, and has a higher gelation temperature, thereby being easily injected. Further, the polymer of the present invention may increase its molecular weight by forming the multi-block of PEO-PPO (or PBO)-PEO units, thereby maintain the gel state for a long period in the body or aqueous solution. Therefore, when the polymer of the present invention is used as a drug delivery system, the drug may be continuously released for 24 hours or more with one injection, thereby overcoming the disadvantage of the conventional poloxamer which has a short sustain period.
  • the multi-block copolymer of the present invention is hydrolysable due to an ester bond formed by the carboxylic linker, and the degraded PEO-PPO (or PBO)-PEO unit having a low molecular weight and dicarboxylic acid are water-soluble, thereby being easily eliminated out of the body. Since a degradation rate of the polymer is proportional to the number of the dicarboxylic linkers contained in the polymer, the degradation rate and the size of hydrolysed products may be controlled by a size and number of the blocks.
  • a microsphere produced with the multi-block copolymer of the present invention may be used as a drug delivery system, when the drug is included in the microsphere.
  • the applicable drug may be any drug, for example a nonionic drug or an ionic drug, preferably an ionic drug such as a peptide or a protein containing a large number of carboxylic and amino ionic groups in the molecule.
  • the peptide or the protein may be e growth hormone (GH), interferon (INF), granulocyte colony stimulating factor (G-CSF), erythropoietin (EPO), interleukin (IL), follicle stimulating hormone (FSH), nerve growth factor (NGP), octreotide, insulin, calcitonin, tumor necrosis factor (TNF), vascular endothelial growth factor (VEGF), epithelial growth factor (EGF), platelet derived growth factor (PDGF), bone morphogenic protein (BMP), tissue plasminogen activator (TPA), and oligonucleotide.
  • GH growth hormone
  • INF interferon
  • G-CSF granulocyte colony stimulating factor
  • the protein may be a natural form or a modified form with a polymer such as polyethylene glycol
  • some proteins may be mixed with a metal ion such as Zn 2+ , Ca 2+ , Cu 2+ , Mg 2+ , and the like, to form a complex, leading to delay the release of the drug from a gel.
  • the microsphere of the present invention may include the multi-block copolymer of 40 to 99.9% by weight, and the drug of 0.01 to 60% by weight.
  • the multi-block copolymer solution with the concentration of 0.5 to 50% may be used as a drug delivery system so long as it exhibits the phase transition.
  • an aqueous solution containing the multi-block copolymer of the present invention Since the polymer of the present invention is easily dissolved at a low temperature of about 4 0 C and not easily dissolved at a room temperature of about 25 0 C, it is preferable to dissolve the polymer at the low temperature.
  • An amount of the multi-block copolymer capable of being dissolved may be limited according to a molecular weight. When the molecular weight of the multi-block copolymer is about 100,000, it may be dissolved in water with the maximum concentration of 30%, preferably with the concentration of 4 to 20%.
  • the aqueous solution becomes a gel state at the body temperature, thereby releasing slowly the peptide, the protein, or the water-soluble drug included therein.
  • EXAMPLE 2 Synthesis of a poloxamer oligomer using oxalyl chloride as a linker
  • EXAMPLE 1 The same method was used as EXAMPLE 1 except using oxalic chloride as a dicarboxylic linker.
  • the molecular weight of an obtained poloxamer oligomer was 91,300.
  • EXAMPLE 1 The same method was used as EXAMPLE 1 except using adipoyl dichloride as a dicarboxylic linker.
  • the molecular weight of an obtained poloxamer oligomer was 96,300.
  • EXAMPLE 4 Synthesis of a poloxamer oligomer using suberoyl dichloride as a linker
  • EXAMPLE 1 The same method was used as EXAMPLE 1 except using suberoyl dichloride as a dicarboxylic linker.
  • the molecular weight of an obtained poloxamer oligomer was 97,800.
  • EXAMPLE 5 Synthesis of a poloxamer oligomer using sebacoyl dichloride as a linker The same method was used as EXAMPLE 1 except using sebacoyl dichloride as a dicarboxylic linker. The molecular weight of an obtained poloxamer oligomer was 124,000.
  • EXAMPLE 6 Synthesis of a poloxamer oligomer using dodecandioyl dichloride as a linker
  • EXAMPLE 7 Synthesis of a poloxamer oligomer using terephthaloyl dichloride as a linker
  • EXAMPLE 1 The same method was used as EXAMPLE 1 except using terephthaloyl dichloride as a dicarboxylic linker.
  • the molecular weight of an obtained poloxamer oligomer was 87,000.
  • EXAMPLE 8 Synthesis of a poloxamer oligomer using fu malic acid as a linker 1Og of fumaric acid and 22g of oxalic chloride (2 equivalents based on 1 equivalent of fumaric acid) were quantified and added in 5OmL of acetonitrile, and reacted for 6 hours at 5O 0 C. After reaction was completed, excess oxalyl chloride was removed in vacuum condition, thereby obtaining reactive fumaroyl dichloride. The synthesized fumaroyl dichloride was used as a dicarboxylic linker, and the same method was used as EXAMPLE 1 to synthesize a poloxamer oligomer. The molecular weight of the obtained poloxamer oligomer was 85,400.
  • EXAMPLE 9 Synthesis of a poloxamer oligomer using maleic acid as a linker The same method was used as EXAMPLE 8 except using maleic acid. The molecular weight of an obtained poloxamer oligomer was 82,700.
  • EXAMPLE 10 Synthesis of a poloxamer oligomer using malic acid as a linker The same method was used as EXAMPLE 8 except using malic acid. The molecular weight of an obtained poloxamer oligomer was 84,000.
  • EXAMPLE 11 preparing a microsphere using a multi-block copolymer solution 700 mg of the multi-block copolymer prepared in EXAMPLE 1 (molecular weight: 101,500) was dissolved in 9.3g of distilled water, and the solution was left at 4 0 C for 2 hours forming 7% multi-block copolymer solution.
  • the multi-block copolymer solution was injected through a 0.7mm nozzle with maintaining a temperature at 1O 0 C, thereby being contacted with liquid nitrogen. Subsequently, the temperature of the reaction chamber was maintained at -2O 0 C, and freeze-drying is performed for 6 hours using a vacuum pump, to prepare a microsphere.
  • the microsphere was observed through an electron microscope, and a result is shown in FIG. 3. As shown in FIG. 3, the size of the microsphere is within a range of 10 to 200 ⁇ m.
  • EXAMPLE 12 preparing a microsphere using a multi-block copolymer solution mixed with an interferon 1.28mg of interferon-alpha (Sigma®) was dissolved in 1OmL of distilled water, and 1.46mL of zinc acetate (lmg/mL) was added thereto so that the mole ratio between Zn and interferon would be 100:1, to form interferon-Zn complex.
  • 840mg of the multi- block copolymer prepared in EXAMPLE 1 was added to the solution, and the solution was left at 4 0 C for 3 hours, thereby forming 7% interferon-contained multi-block copolymer solution.
  • a microsphere is manufactured by the same method as EXAMPLE 11 using the interferon-multi-block copolymer solution. The size of the obtained interferon-contained multi-block copolymer microsphere is within a range of
  • EXAMPLE 13 preparing a microsphere using a multi-block copolymer solution mixed with a hyaluronic acid
  • EXAMPLE 13 preparing a microsphere using a multi-block copolymer solution mixed with a human growth hormone
  • HGH human growth hormone

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Abstract

L'invention concerne un appareil et un procédé de préparation de microsphères de polymère. Le procédé comprend les étapes suivantes : injection d'une solution d'un polymère hydrosoluble biocompatible, à haut poids moléculaire, à travers une tuyère, sous une pression de l'ordre de 0,5 bar à 15 bar, de manière à former des gouttelettes, congélation des gouttelettes en mettant en contact ces gouttelettes avec un gaz liquéfié, de manière à former des particules congelées ; retrait du gaz liquéfié ; et lyophilisation des particules congelées à une température de l'ordre de -70 °C à 1 °C et sous une pression égale ou inférieure à 1 torr.
PCT/KR2006/004263 2005-12-20 2006-10-19 Procede de preparation de microspheres de polymere, et appareil de fabrication correspondant WO2007073035A1 (fr)

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WO2009012449A1 (fr) * 2007-07-18 2009-01-22 The Board Of Trustees Of The University Of Illinois Libération temporaire de facteurs de croissance à partir d'échafaudages de micro-tige en 3d pour régénération de tissu
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