KR20110022096A - Method for preparing microspheres of polymer and manufacturing apparatus therof - Google Patents

Abstract

PURPOSE: A method and apparatus for preparing polymer microspheres are provided to prevent denaturation and activity reduction of physiologically active drug of proteins. CONSTITUTION: A method for preparing polymer microspheres comprises: a step of spraying biocompatible soluble polymer solution through a nozzle at 0.5-15 bar of pressure to form a droplet; a step of contacting the droplet with liquefied vapor to form a frozen particle; and a step of removing liquefied vapor and freeze-drying the particles at -70°C to -1°C. The biocompatible soluble polymer is hyaluronic acid, dextran, gelatin, collagen, chitosan, methyl cellulose, ethyl cellulose, hydroxy ethyl cellulose, or hydroxy propyl methyl cellulose. The liquefied vapor is liquefied nitrogen, liquefied oxygen, or liquefied helium.

Description

TECHNICAL FOR PREPARING MICROSPHERES OF POLYMER AND MANUFACTURING APPARATUS THEROF

The present invention relates to a method and apparatus for producing microspheres using biocompatible polymers. More specifically, spraying the biocompatible polymer solution at a constant pressure through a nozzle to form a droplet; Contacting the formed droplets with a liquefied gas to form frozen particles; And removing the liquefied gas and lyophilizing the frozen particles to prepare a polymer microsphere. The present invention is characterized in that it is carried out at low temperature using a biocompatible polymer that is soluble in water, without using an organic solvent.

Over the past 30 years, microspheres have been studied as one of the drug delivery systems. Among them, in particular, it is applied to a drug that is effective only after repeated administration, and the prostate cancer treatment agent was developed first. In the case of prostate cancer, LHRH, which is a type of sex hormone, should be periodically administered to inhibit cancer growth, and a microsphere formulation has been developed in which frequent administration is very inconvenient for patients, and can be administered more than one month as a single administration. .

As a method for preparing microspheres, solvent evaporation drying method, phase separation method, spray drying method, low temperature solvent extraction method and the like are known, but most solvents for dissolving biodegradable polymers, such as dichloromethane or chloroform, are harmful to the human body.

Y Ogawa, et al., Chem. Pharm. Bull. 36 2576-2581 (1988) introduced a process for preparing Leuprorlelin acetate depot formualtion using polymers. Leuprorelin acetate is dissolved in an aqueous gelatin solution, which is then mixed with a methylene chloride solution containing polylactic glycolic acid to form a W / O emulsion. The mixed emulsion was dropped in 0.25% PVA aqueous solution to form a W / O / W emulsion, followed by removing the organic solvent methylene chloride and freeze-drying to remove the remaining water. The microspheres thus prepared sustained release of leuprorenin acetate for at least one month in a single injection. LHRH had no difficulty in preparing microspheres from peptides with a molecular weight of 1100, but protein drugs with higher molecular weights were more likely to lose protein-specific activity in response to organic solvents or temperature changes during microfabrication.

Jeffery L. Cleland et al., Journal of controlled Release 49 (1997) describe a human growth hormone microsphere as a cryogenic process that is a new method of improving human growth hormone (hGH) to existing W / O / W methods. Was prepared. This human growth hormone microsphere slowly released human growth hormone which has activity for one month after administration to the body.

U. S. Patent No. 5922253 describes a device and a manufacturing method related to a cryogenic process, which is one of processes for manufacturing human growth hormone microspheres. After dissolving polylactic glycolic acid (PLGA) or polylactic acid (PLA) in an organic solvent to make microspheres, the mixture is sprinkled with a liquid gas and kept at a low temperature to freeze. The droplets of were precipitated in a solvent that did not dissolve polylactic glycolic acid (PLGA) or polylactic acid (PLA) to extract the initial organic solvent. This manufacturing process has a number of advantages, the yield is high, it was possible to produce commercially available microparticles (microparticles). It was also possible to work under sterile conditions and to control the size of the microparticles. However, because organic solvents were used in this process, there was a risk of protein denaturation. Peptides or protein drugs decrease activity due to a number of factors, which are more severe when exposed to contact with organic solvents, pH changes, and shear forces.

In order to eliminate the problems caused by such organic solvents, several different types of protein delivery systems have been studied, a typical temperature-sensitive polymer hydrogel. The polymer used in this formulation is a substance that causes the phase change of sol-gel when dissolved in water. It is liquid at low temperature and can be mixed with protein or peptide drug. It is a system that turns into a solid and slowly releases proteins or peptides. In such formulations, the decrease in activity can be reduced to a minimum because no organic solvent is used that reduces the activity of the protein. A commonly studied material for such hydrogel systems is poloxamer 407, a polymer. When the polymer is a block copolymer of polyethylene glycol (PEG) and polypropylene glycol (PPO), when dissolved in water to form an aqueous solution, the polymer is in a liquid state at low temperature, and then turns into a solid gel as the temperature increases. These polymers are called temperature sensitive polymers, and research into using the polymer solution as a carrier of proteins or peptides has continued.

However, in the case of the poloxamer 407, there must be at least 25% of a polymer solution in order to have a sustained release effect in the body, and moreover, in a real animal experiment, most protein or peptide drugs are released after 1 day of administration. It wasn't.

U.S. Patent No. 6004573 describes a block copolymer made of PEG and PLGA, which is a copolymer having a composition of PEG-PLGA-PEG, which is similar to poloxamer 407 when dissolved in water. That is, it exists as a sol in a liquid state at low temperature, but has a characteristic of changing into a gel when the temperature rises. The PEG-PLGA-PEG copolymer was studied in a formulation that releases paclitaxel, an anticancer agent that is insoluble in water for one month. A solution of PEG-PLGA-PEG copolymer having a sol-gel phase transition phenomenon can be used as a protein drug carrier. Since it is in a sol state at a low temperature, it can be easily mixed with a protein. Can be released slowly.

A polymer composition composed of poloxamer 407 that is degradable in vivo is known from X. Zhao et al. In "controlled release society 30th annual meeting Proceedings # 167". X. Zhao et al. Disclose a compound in which Ploxamer 407 is linked to a degradable carbonate bond by reacting Ploxamer 407 with DSC (disuccinimidyl carbonate), and US Pat. No. 6,348,558 discloses two or more compounds by hydrolyzable carbonate linkages. Degradable polymers linked by polyalkylene oxide oligomers are known.

Therefore, there is a demand for applying a microsphere that does not use an organic solvent and a novel polymer therefor.

An object of the present invention is to provide a method for producing microspheres using a polymer solution without using an organic solvent.

In addition, another object of the present invention is to provide a manufacturing apparatus capable of producing microspheres using a polymer solution without using an organic solvent.

The present invention comprises the steps of spraying a biocompatible water-soluble polymer solution at a constant pressure through a nozzle to form a droplet; Contacting the formed droplets with a liquefied gas to form frozen particles; And removing the liquefied gas and lyophilizing the frozen particles to produce microspheres, and a manufacturing apparatus for use therein.

The present invention is characterized in that the microspheres are prepared without using an organic solvent by freezing and lyophilizing the droplets generated by spraying the biocompatible polymer solution. The biocompatible polymer is preferably a water-soluble polymer, and preferably has a high molecular weight, more preferably, a sol-gel phase transition characteristic of forming a sol state at a low temperature and forming a gel state at a high temperature. In particular, it is preferable that it is the multiblock polymer mentioned below.

The terms used in the present invention are summarized as follows.

"Biodegradable or biocompatible" means a property that is administered into the body and then hydrolyzed or degraded by enzymes to be absorbed or safely released into the body.

"Microsphere" refers to microspheres up to 1 mm in size, which means that they are spherical in shape in a series of microparticles, commonly called microparticles, and may range from several micrometers to several hundred micrometers in size. will be.

"Poloxamer" is a compound in which a hydrophilic block of polyethylene oxide (PEO) and a hydrophobic block of polypropylleoxide (PPO) are linked by ether bonds in a triple block of PEO-PPO-PEO type, and the weight average molecular weight of the polymer It refers to a block copolymer of 1,000 Daltons to 20,000 Daltons, both terminal group is a hydroxyl group, in particular poloxamer 188 (Fluronic ^® F-68) and poloxamer 407 (Fluronice ^® F-127) commercially available compounds Can be.

A sol-gel phase transition exists in a sol state below a certain temperature and then turns into a gel state when the temperature rises above a certain temperature, and in a gel state above a certain temperature and then falls below a certain temperature. It means to change to a sol state, which is a reversible reaction. The specific temperature is difficult to specify because it changes depending on the type of polymer, the concentration of the aqueous solution, the presence of salt, the hydrogen ion concentration, etc., the specific temperature range may be a specific temperature between 5 to 37 ℃, preferably, 25 To a specific temperature in the range from 35 ° C.

"Freeze-drying" is a method in which various substances are generally dissolved in water, frozen at 0 ° C. or lower, and decompressed using a vacuum pump to cause sublimation of water used as a solvent, followed by drying.

First, the present invention provides a method for preparing microspheres by spraying a polymer solution at a constant pressure through a nozzle to form droplets, freezing and lyophilizing them.

More specifically, the manufacturing method of the present invention,

Spraying the biocompatible water-soluble polymer solution at a constant pressure through a nozzle to form droplets;

Contacting the formed droplets with a liquefied gas to form frozen particles;

Removing the liquefied gas and lyophilizing the frozen particles to prepare a polymer microsphere.

Preferably the present invention,

Spraying a high molecular weight biocompatible water soluble polymer solution through a nozzle at a pressure ranging from 0.5 to 15 bar to form droplets;

The formed droplets are contacted with the liquefied gas and frozen to form frozen particles;

Removing the liquefied gas and freeze drying the frozen particles at a temperature in the range of −70 ° C. to −1 ° C. and a pressure of 1 Torr or less,

Provided is a method for producing a polymeric microsphere.

In the present invention, a droplet is formed by spraying a polymer solution at a predetermined pressure in a range below a predetermined temperature through a nozzle, and the resulting droplet is contacted with a liquid gas to freeze particles having a size of several micrometers to several hundred micrometers. Create Then, the liquefied gas is removed, and the freeze particles are lyophilized by decompression at low temperature to prepare a polymer microsphere.

The present invention also provides an apparatus for producing a polymer microsphere comprising a reaction vessel, an injection unit and a temperature control unit.

Specifically, as a manufacturing apparatus for producing microspheres from a biocompatible polymer solution,

A reaction vessel in communication with the vacuum pump through the negative pressure pipe, the inside of which is formed as a vacuum, and the gas generated therein can be exhausted through the exhaust port, and an inclined surface inclined around the outlet port is formed on the inner bottom surface;

The nozzle includes a nozzle extending inwardly through the reaction vessel, the nozzle is in communication with the pressure control pump to spray the polymer solution supplied at a constant pressure into the reaction vessel to form a droplet of uniform shape Injection unit to be;

And a container side heat exchange means installed in a space formed on the wall of the reaction vessel 2, and nozzle side heat exchange means surrounding a portion exposed from the nozzle to the outside of the reaction vessel. A temperature control unit which circulates the heat exchange medium that is pumped by to maintain the internal atmosphere of the reaction vessel and the periphery of the nozzle at a predetermined temperature or less so that the polymer solution is not gelled;

It provides a microsphere manufacturing apparatus equipped with.

In an embodiment of the present invention, the configuration of a device suitable for the production of the polymer microsphere may be that shown in Figs.

Hereinafter, the configuration of the manufacturing apparatus illustrated in FIGS. 1 and 2 and the microsphere manufacturing method using the same will be described in detail.

1 and 2, the microsphere manufacturing apparatus according to the present invention is disposed in the reaction vessel 2 and the outer side of the reaction vessel 2, the injection portion 4 for injecting a polymer solution therein And it is composed of a temperature control unit 6 for keeping the internal temperature of the reaction vessel 2 within a certain range.

The injection unit 4 disposed at the upper center or side of the reaction vessel 2 includes a nozzle 40 having a tip extending therein through the reaction vessel 2, and a polymer injected through the nozzle 40. It is composed of a pressure regulating pump 42 to supply a solution at a constant pressure to form a droplet of a uniform shape inside the reaction vessel (2). The reaction vessel 2 may be preferably a cylindrical volume of 20L to 30L, the material of the reaction vessel may be of a material capable of withstanding a vacuum of 1 torr or less, preferably steel or stainless steel Can be. Since the diameter of the nozzle 40 affects the size of the sprayed droplets, the diameter of the nozzle 40 may be variously adjusted according to the desired droplet size, and preferably, 0.1 to 1 mm. In addition, a predetermined space may be formed between a wall of the reaction vessel, that is, an inner surface and an outer surface of the wall, and a heat exchange medium may exist in the space.

The nozzle 40 may be a nozzle and an atomizer used in a general spray-dryer. Generally available atomizers may be external air atomizers or internal air atomizers and the nozzles may be spray-dryers (bucci).

In addition, the temperature control part 6 is a jacket-side pipe-side heat exchange means 60 is installed in the space formed in the wall of the reaction vessel (2) and the outside of the reaction vessel (2) in the nozzle (40) And a nozzle-side heat exchange means 62 in the form of a pipe that surrounds the exposed surroundings, and these heat exchange means 60 and 62 are provided by a temperature controller 64 separately provided from the reaction vessel 2. The heat exchange medium is circulated to supply liquid to remove the liquefied gas and maintain the inner atmosphere of the reaction vessel 2 and the periphery of the nozzle 40 in the range of -40 to -5 ° C to prevent gelation of the polymer solution or droplets. have.

The temperature regulating device 64 may employ a conventional cooling device, and in this case, a heat exchange medium, that is, a CFC-based solvent such as HCFC22, HFC134a, R410a, and R407c may be used.

In addition, when the droplet of the polymer solution is in contact with the liquefied gas in the lower side of the reaction vessel (2) to freeze, when the problem that the condensed spherical body, that is, frozen particles are entangled with each other occurs, the stirring blade 32 is operated to solve the problem. can do. In addition, it may further include means such as a motor and a speed reducer (not shown) for rotating the stirring blade (32). The stirring blade may be a propeller type, a furnace type, a turbine type, a spiral weight type, an Or type or an impeller type, and in one embodiment of the present invention, may be a propeller type.

In addition, the reaction vessel (2) may be implemented in a structure that can be opened and closed by using the upper portion as a cover, in which case the cover must be provided with a gasket or packing for airtight. This is to remove the air inside the container by using a pressure control pump, etc., to adjust the pressure, so as to block the penetration of external air to increase the degree of vacuum inside the container.

In addition, in a structure in which a lid is added to the reaction vessel 2, a handle 34 for simplifying the removal and assembly of the lid may be attached to an appropriate position of the reaction vessel 2.

The gas exhaust port 24 through which the vaporized liquefied gas is discharged may be positioned above the reaction vessel 2. The exhaust port 24 freezes the polymer solution droplets injected by the liquefied gas to form frozen particles, and then, when the liquefied gas is removed, the vaporized liquefied gas is discharged. By removing the gas generated in contact with the liquefied gas, a large amount of droplets when the contact with the liquefied gas is generated a large amount of gas also serves to prevent the increase in the internal pressure of the reaction vessel (2).

At the bottom of the reaction vessel 2, there is an outlet 26 for collecting the microspheres generated by the lyophilization and freezing of the droplets of the polymer solution. In order to facilitate the collection of the generated microspheres, the bottom of the storage space of the heat exchange medium may be manufactured to be inclined.

In the microsphere apparatus having the above configuration, the reaction vessel 2 is formed in a vacuum state of 1 torr or less by the negative pressure action of the vacuum pump 20 through the negative pressure pipe 22, and the anti-container ( 2) The inside of the inside is to start the manufacture of microspheres in a state where the liquefied gas is filled to a certain level.

In the microsphere manufacturing method of the present invention, first, the polymer solution is sprayed through a nozzle 40 at a constant pressure to perform a step of forming droplets.

The injection of the polymer solution is classified into a direct injection method and an indirect injection method according to the position of the injection nozzle. In the direct injection method, as shown in FIG. 1, the nozzle 40 is positioned above the reaction vessel 2, and when the polymer solution G1 is injected by pressure, the sprayed droplet moves in the same direction as gravity, It is rapidly contacted with the liquefied gas located to freeze to form condensed globules (G2), that is, frozen particles. According to this method, a large amount of aqueous solution can be frozen in a short time, but if it is not sufficiently stirred, there is a possibility that the droplets become entangled upon contact with the liquefied gas.

Indirect injection method is the injection nozzle is located on the side of the container (Fig. 2). Indirect injection method is sprayed by lowering the pressure than the direct injection method, the polymer solution (G1) injected from the nozzle 40 proceeds in the direction of gravity and vertical horizontally with the nozzle 40, the reaction vessel (2) by gravity As it falls down and freezes in contact with the liquefied gas, condensed globules G2, that is, frozen particles are formed. With this method, the freezing process will occur slowly, resulting in relatively small droplets, but will take a long time to freeze the droplets.

As the water-soluble polymer of the present invention, all synthetic or natural water-soluble polymers can be used, and those having a high molecular weight are most preferred, and those having sol-gel phase transition properties are most preferable. The polymer of the present invention is hyaluronic acid (Hyaluronic acid), dextran, gelatin, collagen, chitosan, methyl cellulose (MC), ethyl cellulose (EC), hydroxyethyl cellulose (HEC), methyl hydroxyethyl cellulose ( MHEC), hydroxymethylcellulose, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose and triplet with polypropylene oxide (PPO) or polybutylene oxide (PBO) between two polyethylene oxide (PEO) blocks The block copolymer may be selected from the group consisting of polymers multiblocked by a dicarboxyl linker.

In the method of the present invention, the higher the molecular weight of the polymer used, or the higher the concentration of the aqueous solution, it is possible to produce a microsphere having a dense structure. The polymer of the present invention may have a weight average molecular weight of 25,000 Daltons to 1,000,000 Daltons, preferably 40,000 Daltons to 1,000,000 Daltons. Microspheres made of low molecular weight polymers, such as poloxamers, have a problem of dissolving before re-injection after reconstitution in aqueous solution, but high molecular weight polymers have a high strength of the prepared microspheres, thus maintaining the shape of the microspheres in the body. It is possible to inject. In addition, low-molecular weight polymers are liable to be broken during freeze-drying, so the recovery rate may drop. However, the high-molecular-weight polymers of the present invention are highly preferable because they retain their elasticity even after freeze-drying, so that the low-molecular weight is low.

The polymer solution is obtained by dissolving a water-soluble polymer in an aqueous solvent such as distilled water, acetate buffer solution, phosphate buffer solution, peptide or protein solution, and the concentration of the polymer solution is 3 to 25% (w / w), more preferably 3 To 15% (w / w). When the concentration of the polymer solution is lower than the above range, the particle formation is not good, and when the concentration of the polymer solution is above the above range, excessively high pressure is required at the time of spraying, which is not preferable.

The water-soluble polymer solution used in the present invention is most preferably in a form capable of sol-gel phase transition, and is an excellent drug delivery material capable of sustained release of drugs and the like when the drug is encapsulated and administered into the body. It is preferable that the water-soluble polymer having the sol-gel phase transition property is the following multiblock polymer.

Due to the sol-gel phase transition property of such a water-soluble polymer, preferably, a multi-block polymer solution, when the temperature of the solution becomes higher than the sol-gel phase transition temperature, it becomes difficult to form droplets in the gel state. The spraying temperature of the solution is preferably kept below the sol-gel phase transition temperature. As such, the spraying temperature of the polymer solution must be maintained below the sol-gel phase transition temperature to lower the viscosity of the solution to prepare the microspheres. In the present invention, the temperature of the multi-block polymer solution during spraying is preferably maintained at 15 ° C. or less, and more preferably the temperature can be maintained in the range of 5 to 12 ° C. At this time, a normal nozzle side heat exchange means 62 is also installed around the nozzle 40, and when the polymer solution G1 has a sol-gel phase transition characteristic, the temperature of the aqueous solution is maintained at a predetermined temperature or lower, Gel formation due to temperature rise can be prevented.

In the present invention, the polymer solution is injected through the nozzle 40 by a constant pressure applied by the pressure control pump 42, the larger the injection pressure is the size of the droplets generated by the injection is reduced, by adjusting the injection pressure size The size of the generated droplets can be adjusted, and also the injection speed can be adjusted by adjusting the injection pressure. The preferred injection pressure range is determined in consideration of the size of the droplet to be produced, the injection speed, and the like, and is generally in the range of about 0.5 to 15 bar, preferably 1 to 5 bar. If the injection pressure is lower than the above range, the injection of the aqueous solution does not occur, or the size of the resulting droplets is large, it is not good to take a long time to the injection, if it is higher than the above range, the injection rate is too fast to entangle the generated droplets is preferable Not.

The sprayed and dropleted polymer solution (G1) falls into the lower portion of the reaction vessel (2) and rapidly freezes as it comes into contact with the liquefied gas and converts into spherical body (G2), that is, frozen particles.

The droplets of the polymer solution injected from the nozzle 40 are rapidly frozen as soon as they come into contact with the liquefied gas and form particles in the size of the droplets formed during the injection. Therefore, the particle size may be adjusted by adjusting the size of the droplets formed during the spraying, and the size of the droplets may be adjusted by the spray nozzle diameter and / or the spraying pressure.

It is preferable to stir the contact state between the droplet and the liquefied gas in order to prevent the droplets from being entangled when frozen particles are formed by the freezing of the droplet by contact with the droplet and the liquefied gas. At this time, if the stirring speed is too fast, the frozen particles (G2) of the droplets stick together, and if it is too slow, the droplets stick together to form agglomerates before freezing, which is not preferable. Depending on the method can maintain the proper stirring speed. Preferred stirring speed is about 5 to 300 rpm, the stirring speed may vary depending on the viscosity, concentration, process of the solution of the polymer solution to be injected. In the present invention, more preferably, at the time of injection of the polymer solution, the stirring may be performed at about 50 to 100 rpm, and after the injection is completed, the stirring may be performed at the speed of about 60 to 150 rpm.

The liquefied gas usable in the present invention is not limited, but the boiling point is preferably about -20 ° C. or lower for rapid freezing of the droplets, and more preferably selected from the group consisting of liquefied nitrogen, liquefied oxygen, and liquefied helium.

In addition, the manufacturing method of the present invention includes the step of lyophilizing after removing the liquefied gas from the frozen droplets, that is, frozen particles (G2). Specifically, the condensed spherical body G2, that is, the lyophilized particles, is lyophilized under lyophilized conditions formed by the container-side heat exchange means 60, the temperature controller 64, and the vacuum pump 20 to form a porous sphere. The microspheres are formed, and the formed microspheres are collected by rolling off the inclined surface 28 at the bottom of the reaction vessel 2 through the outlet 26.

After the spraying of all the water-soluble polymer solutions, the temperature controller 64 connected to the heat exchange means 60 at the bottom of the reaction vessel 2 is operated to raise the temperature of the reaction vessel 2 below the boiling point of the liquefied gas. To evaporate and remove the liquefied gas. The vaporized liquefied gas is removed by the exhaust port 26 above the reaction vessel 2. At this time, the temperature of the lower portion of the reaction vessel (2) should be above the boiling point of the liquefied gas, it can be maintained at about -70 to -1 ℃, preferably -40 to -5 ℃, more preferably about -20 ℃. .

After removing the liquefied gas, the temperature of the lower portion of the reaction vessel 2 for freeze-drying the frozen particles is maintained at -70 to -1 ° C, preferably -40 to -5 ° C, and more preferably about -20 ° C. The pressure in the vessel 2 is preferably lowered to 1 torr or lower by operating the vacuum pump 20.

At this temperature and pressure, the water contained in the frozen particles is dried while subliming to produce spherical microspheres having pores formed by the sublimation of water. In order to reduce the pressure in the container with the vacuum pump 20, preferably, all the entrances and exits 26 attached to the container, the outlet 26, the inlet 26 for collecting the manufactured microspheres, and the injection connected to the nozzle 40 The part 4 etc. can be sealed completely.

When preparing the microspheres by freeze-drying the frozen particles (G2), as described above, the temperature can be maintained at -70 to -1 ℃, preferably, -40 to -5 ℃, preferably pressure It can be kept below 1 torr. If the temperature and pressure inside the reaction vessel 2 is above the above range, the frozen particles are aggregated or melted so that they do not maintain a spherical shape. Freeze drying may be performed using the vacuum pump 20 for about 6 hours to 12 hours, and the temperature is about -70 to -1 ° C, preferably -40 to -5 ° C, and more preferably, up to about 3 hours. It can dry, maintaining about -20 degreeC, and after that, it can dry, gradually raising to room temperature.

The size of the prepared microsphere can be adjusted by the diameter of the nozzle 40 and the pressure applied during the injection. The microspheres prepared by the above method preferably have a size of about 10 μm to 500 μm, and preferably have a porous surface. The pores on the surface are caused by the sublimation of water contained in the polymer solution during freeze drying.

In the preparation of the microsphere of the present invention, a bioactive substance may be additionally added to the biocompatible polymer solution. Drugs that can be used include protein drugs or peptide drugs, and include various protein or peptide hormones, various antibiotics, growth hormone (GH), interferon (IFN), leukocyte proliferation factor (G-CSF), erythrocyte producing protein ( EPO), interleukin (IL), follicle stimulating hormone (FSH), nerve growth factor (NGF), octreotide, insulin, calcitonin, tissue necrosis factor (TNF), angiogenesis factor (VEGF), epithelial growth factor ( EGF), platelet growth factor (PDGF), bone morphogenetic factor (BMP), thrombolytics (TPA) and oligonucleotides may be selected from the group consisting of. In this case, the protein or peptide may be included in a buffer solution, and when the biocompatible polymer is lyophilized by dissolving the biocompatible polymer in a buffer solution containing the protein or peptide, the microspheres containing the protein or peptide do not have water. Therefore, the microspheres containing the protein or peptide can be stored for a long time.

The content of the protein or peptide drug may be 0.01 to 50% by weight based on the weight of the polymer, which can be adjusted in consideration of the release time, release form, etc., to maintain the manufacturing characteristics or form of the microspheres properly For this reason, it is preferable that they are 0.1 weight%-20 weight%.

The microspheres of the present invention may be used as such or in nanosphere form, or may be prepared in a mixed sol-gel depot-type drug carrier, strip, rod, film form, or the like as a sustained-release drug carrier.

In addition, the microsphere of the present invention, PEG, hyaluronic acid, dextran, gelatin, collagen, chitosan, poloxamer 407, poloxamer 188, methyl cellulose (MC), ethyl A polymer selected from the group consisting of cellulose (EC), hydroxyethyl cellulose (HEC), methyl hydroxyethyl cellulose (MHEC), hydroxy methyl cellulose, hydroxypropyl methyl cellulose (HPMC) and hydroxypropyl cellulose It may further comprise. The polymer may be further included in an amount of 0.1 to 50% by weight, based on the weight of the polymer in the polymer solution. In this case, the microspheres of the present invention may be prepared in the form of mixed sol-gel depot-type drug carriers, microspheres, nanospheres, strips, rods, films, and the like, and may be used as mixed sustained release drug carriers. When the mixed drug carrier is prepared, the gelation temperature of the polymer of the present invention or the strength of the gel may be changed.

When preparing microspheres using polymer solutions containing protein or peptide drugs, stabilizers and / or various excipients for stabilizing protein or peptide drugs may be added. The stabilizers and excipients keep the protein or peptide drug active in the polymer solution without denaturation, and act as a buffer during the lyophilization process in the preparation of microspheres to have the inherent activity of the protein or peptide drug.

As the stabilizer, PBS (Phosphate Buffer Solution), amino acids, carbohydrates, fatty acids, surfactants, etc. may be mainly used, and excipients may include lactose, dextran, mannitol, sucrose, and the like. When adding stabilizers and / or excipients the amount of addition depends on the properties of the desired microspheres. When the excess stabilizer and / or excipient is added, the physical properties of the microspheres change, so the content of the stabilizer and / or excipient must be maintained in a certain range, and it is appropriate that the amount is about 1% to 20% by weight based on the polymer weight. Do.

The addition of metal cations to some proteins or peptides may result in higher stability and control the release rate in the lyophilized state. The amount of metal cation added depends on the type of protein or peptide, and may be mixed in a protein or peptide to metal cation molar ratio of about 1:10 to 1: 5000 (moles of protein or peptide: moles of metal cations). This is the metal cation is preferably represent the same effect may be ^{Ca 2+, Zn 2 +, Mg} 2 +.

The administration method and dosage of the drug delivery vehicle are determined according to the pharmacological activity of the contained drug, the site of action in the body, the physicochemical properties, and the like. For example, the administration method may be, for example, oral administration, subcutaneous administration, intravenous administration, intraperitoneal administration, and the like.

Conventional microspheres are prepared by dissolving polylactic glycolic acid (PLGA) or polylactic acid (PLA) in an organic solvent and dropping them in an aqueous solution to form spherical droplets and removing the organic solvent. Instead of using an organic solvent, the present invention dissolves and sprays a biocompatible polymer such as a multiblock polymer in an aqueous solvent to form droplets and uses a liquefied gas so that it is not harmful to the human body and contains bioactive drugs such as proteins. It can also increase the stability by preventing its degeneration and reduced activity.

In addition, the biocompatible polymer of the present invention may be dissolved in an aqueous solvent such as water and then mixed with a physiologically active drug such as a protein or a peptide to prepare a sustained-release depot formulation. The syringe containing the polymer solution must be kept cold until the preparation is injected into the syringe and administered into the body. However, when the administration time is longer or the external temperature is higher than 25 ℃, the sol state of the syringe turns into a solid state, making it difficult to administer. However, when prepared and administered in microspheres, the problem of phase change occurring before administration can be avoided.

When preparing a depot formulation using the multi-block polymer of the present invention, the depot formulation may contain water and may be degraded, thus making it difficult to store for a long time. In addition, even if the freezing to be stored in the form of microspheres do not freeze it is hard to thaw if it is frozen, but in the case of the microsphere formulation of the present invention does not contain water and small size can solve the above problems .

The biocompatible polymer of the present invention is most preferably the following multiblock polymer.

The multi-block polymer is an ionic polymer which forms a sol at low temperature and forms a gel at high temperature, and has a triple block air having polypropylene oxide (PPO) or polybutylene oxide (PBO) between two polyethylene oxide (PEO) blocks. The coalescence is a polymer multiblocked by a dicarboxyl linker.

The multiblock polymer of the present invention forms a hydrogel at a specific concentration and a specific temperature, exhibits a sol-gel phase transition phenomenon, and is biodegradable. In addition, the multi-block polymer is multiblocked by a dicarboxyl group with a linker, the molecular weight is increased, showing an improved gel persistence. In addition, since the multiblock polymer has ionic end groups, it has an effect of delaying drug release from the gel.

In an embodiment of the present invention, the multiblock polymer may be represented by the following Chemical Formula 1.

In said formula, PEO is polyethylene oxide,

Y means polypropylene oxide (PPO), polybutylene oxide (PBO), or a combination of PPO and PBO,

X is -H, or-anion group,

n is an integer between 1 and 100, preferably n is an integer between 3 and 100,

R is-(CH2) _m -or C _{m '} aryl,

m is an integer between 0 and 20, m 'is an integer between 6 and 12,

When X is not H, 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, X When is H, M is not present.

Preferably, the polymer of the present invention may be represented by the formula (2).

In said formula, PEO is polyethylene oxide,

Y means polypropylene oxide (PPO), polybutylene oxide (PBO), or a combination of PPO and PBO,

X is _{-H, -SO 3 -, -PO 3} 2 -, -C (= O) -RC (= O) -O -

n is an integer between 1 and 100, preferably n is an integer between 3 and 100,

R is (CH2) _m -or aryl of C _{m '} ,

m is an integer between 0 and 20, m 'is an integer between 6 and 12,

When X is not H, M is H, or a monovalent or divalent cationic group, preferably a group consisting of Li, Na, K, Ag, Au, Ca, Mg, Zn, Fe, Cu, Co, and Ni M is not present when X is H.

In the polymer, the polyethylene oxide block may include about 2 to 2000, preferably about 5 to 500, and more preferably about 80 to 120 ethylene oxide, and constitutes two PEO blocks in Chemical Formulas 1 and 2. The number of ethylene oxides may be the same or different

The polypropylene oxide block or polybutylene oxide block may include 2 to 2000, preferably about 20 to 500, more preferably about 30 to 250 units.

In the multiblock polymer of the present invention, at least two PEO-PPO (or PBO) -PEO units are connected, and each unit is PEO-PPO-PEO, PEO-PBO-PEO or PEO- (PPO and PBO combination) ) -PEO.

The multiblock polymer of the present invention may have a weight average molecular weight in the range of 25,000 Daltons to 1,000,000 Daltons, preferably 40,000 Daltons to 1,000,000 Daltons, more preferably a weight average molecular weight of 40,000 to 500,000 Daltons, Most preferably 80,000 to 120,000.

As used herein, a "multiblock polymer" is a polymer in which a polyethylene oxide block is linked to a polypropylene oxide or polybutylene oxide block, and then two or more basic units connected to the polyethylene oxide block are connected by a dicarboxyl linker. Means.

The molecular weight ratio of PEO and PPO or PBO of the multiblock polymer of the present invention may vary from the limit of maintaining the water solubility of the polymer, but may be about 0.2: 1 to about 40: 1, preferably 1: 1 to 7.5. : 1, more preferably 1: 1 to 5: 1, and the PEO block may be included in 10 to 85% by weight, preferably 40% to 85% by weight of the PEO-PPO or PBO-PEO unit.

In the multi-block polymer of the present invention, PEO-PPO (or PBO) -PEO units are linked by a dicarboxyl linker, and the linker is hydrolyzable and hydrolyzable in the body.

As used herein, the term "dicarboxyl linker" means an alkyl or aryl compound having two carboxyl groups in one molecule, such as oxalic acid, malonic acid, succinic acid, adipic acid, and the like, and a substance which is not toxic to the human body is preferable. In particular, alkyl dicarboxylic acids, such as malic acid, oxalic acid, succinic acid, glutaric acid, adipic acid, sebacicolic acid, suberic acid, and dodecanoic acid; Unsaturated dicarboxylic acids such as fumaric acid and maleic acid; And it may be selected from the group consisting of aryl dicarboxylic acid, such as phthalic acid, terephthalic acid.

The dicarboxylic acid linker may be linked by an ester bond with a hydroxy group present at both terminal groups of PEO-PPO (or PBO) -PEO, and the ester bond may be hydrolyzed or enzymatically separated from the carboxylic acid and PEO- in an aqueous solution and in vivo. Decomposes into PPO (or PBO) -PEO units.

The sock end of the multiblock polymer of the present invention is a hydroxyl group or an ionic. Anionic groups of the multi-block polymer terminal of the present invention is _{^{-SO 3 -, -PO 3 2 -}} , -C (= O) -RC (= O) -O - salts, etc. are preferred, and groups corresponding anion is Li, Monovalent, such as Na, k, Ag, Au, or divalent metal cations, such as Ca, Mg, Zn, Fe, Cu, Co, Ni, are possible.

In particular, when the polymer containing an anion group in the sock end complexes with the divalent cation metal, it is possible to maintain the more stable form of the gel state to continue the release of the drug from the gel. In addition, when a polymer containing an anion group is mixed with a cationic drug in an aqueous solution, an ionic salt may be formed to reduce the initial release rate of the drug from the multiblock polymer gel, thereby improving the sustainability of drug release. . In addition, when a divalent metal cation salt such as calcium chloride, zinc chloride or magnesium chloride is added to a solution obtained by mixing the multiblock polymer of the present invention having an anion group at the terminal with a drug having an anion, the divalent metal cation is multiblocked with the anionic group of the drug. An ionic complex is formed between the anionic groups at the polymer end, which can delay the initial release rate of the drug from the gel. Therefore, the polymer of the present invention can be effectively applied as a carrier of nonionic and ionic drugs.

In one embodiment of the invention, the multi-block polymer PEO-PPO (or PBO) -PEO may be a commercialized poloxamer.

Poloxamer is a compound in which a hydrophilic block of polyethylene oxide (PEO) and a hydrophobic block of polypropylene oxide (PPO) are linked by ether bonds in a triple block of PEO-PPO-PEO type and have a weight average molecular weight of 1,000 Daltons to 20,000 Daltons. It is a block copolymer whose both terminal groups are hydroxyl groups. According to the present invention may be used for example poloxamer ^{188 (Pluronic ® F-68)} , poloxamer ^{407 (Pluronic ® F-127)} . Poloxamer may be used without purification or purified to prepare the polymer of the present invention, it is easy to produce a high molecular weight polymer when purified. Purification of the poloxamer can be purified by dissolving in methylene chloride and precipitating in hexane or by layer separation in an n-propanol / H ₂ O solvent as known from US Pat. No. 5,800,711.

Method for producing a multiblock polymer of the present invention is as follows.

Specifically in the present invention

1) reacting the PEO-PPO (or PBO) -PEO with 0.5 to 1.0 equivalents of dicarboxylic acid dichloride of the PEO-PPO (or PBO) -PEO terminal hydroxyl group equivalent slowly for 6 hours or more and reacting for 12 hours or more; 2) adding 0.1 equivalent of PEO-PPO (or PBO) -PEO to the reaction solution to react for at least 2 hours to produce a polymer;

3) precipitating the resulting polymer in an ether solvent and dissolving in methanol;

4) It is composed of the step of precipitating the polymer by slowly adding ether to the methanol solution of the above-dissolved polymer so that the volume mixing ratio of methanol / ether is 1/1 to 1/20, the sock end is a hydroxyl group Phosphorus multiblock polymer is prepared.

In addition, in the present invention

1) reacting the PEO-PPO (or PBO) -PEO with 0.5 to 1.0 equivalents of dicarboxylic acid dichloride of the PEO-PPO (or PBO) -PEO terminal hydroxyl group equivalent slowly for 6 hours or more and reacting for 12 hours or more;

2) adding at least 1 equivalent of dicarboxylic acid dichloride of PEO-PPO (or PBO) -PEO terminal hydroxy equivalent to react for 2 hours or more;

3) precipitating the resulting polymer in an ether solvent and dissolving in methanol;

4) A multi-block polymer including a carboxyl group is prepared at the sock end including the step of precipitating the polymer by slowly adding ether with a volume mixing ratio of methanol / ether of 1/1 to 1/20.

In addition, the polymer containing a carboxyl group in the sock end is dissolved in a solvent mixed with water, for example, acetone, acetonitrile and dioxane, and then neutralized with sodium carbonate or sodium hydrogen carbonate, so that both terminal groups of the multiblock poloxamer are dicarboxylic acid sodium salts. Polymers can be prepared.

In addition, the polymer of which both terminal groups are sodium carboxylate salts is treated with calcium chloride, zinc chloride, magnesium chloride, iron chloride, copper chloride, silver nitrate, potassium chloride, or lithium chloride aqueous solution, and then dialyzed to prepare a polymer having a carboxylate metal salt.

Multiblock polymers whose polymer ends are sulfate or phosphate groups

1) dissolving a multiblock polymer having a hydroxy group in a solvent in a solvent and reacting with sulfate trioxide pyridine complex (C ₅ H ₅ NSO ₃ ) or phosphorus oxychloride (POCl ₃ );

2) It may be prepared by neutralization with sodium carbonate or sodium hydrogen carbonate.

When the sock end of the multiblock polymer is another anion, it may be prepared by a known method using the polymer of the present invention.

In the above reaction, dicarboxylic acid dihalide may be directly reacted as a dicarboxylic group linker. However, when dicarboxylic acid itself is used as a starting material, dicarboxylic acid may be activated by oxalyl chloride. It can be converted to dichloride and used.

The reaction can be carried out directly without a solvent or using a solvent, preferably may be selected from the group consisting of dichloromethane, chloroform, teprahydrofuran, acetonitrile, acetone, toluene and dioxane.

In the present invention, by controlling the reaction temperature and reaction time it is possible to control the rate and degree of polymerization of the polymerization reaction to determine the average molecular weight of the polymer of the present invention. The reaction temperature may vary depending on the boiling point of the reaction solvent used, but 60 to 120 ° C. is preferable, and the reaction time is preferably 12 to 72 hours.

In order to increase the reaction rate of the reaction, tin octoate, zinc chloride, or the like may be used as a catalyst, and the reaction may be performed by using amines such as pyridine, dimethylaminopyridine, imidazole, triethylamine, and the like twice the equivalent of dicarboxylic acid. You can increase the speed. However, in order to increase the purity of the polymer of the present invention, it is preferable not to use a catalyst or amines.

The polymerized polymer can be purified by methods known in the art to which this invention pertains, preferably by purifying by using a solvent in which the reactant is soluble while the product polymer is insoluble.

Meanwhile, the polymer of Structural Formula 1 is characterized in that a sol state is formed at a temperature of 2 to 8 ° C. when dissolved in distilled water at a concentration of 2 to 40% and maintains a gel state at a temperature in a range of 30 to 50 ° C.

Compared to poloxamer, the polymer of the present invention is capable of gel formation at a concentration of about 10% lower than that of poloxamer, thereby reducing the toxicity in the body, and the gelation temperature is higher than that of poloxamer. In addition, the polymer of the present invention can maintain the gel state in vivo and in aqueous solution by increasing the molecular weight by multiblocking the PEO-PPO (or PBO) -PEO unit. Therefore, when the polymer of the present invention is used as a drug carrier, the drug can be continuously released for more than 24 hours in one injection, thereby overcoming the disadvantages of the conventional poloxamer having a short maintenance period.

The multiblock polymer of the present invention is hydrolyzable due to the ester linkage of the dicarboxyl linker, and since PEO-PPO (or PBO) -PEO and dicarboxylic acid having a small molecular weight that is degraded are water soluble, they can be easily discharged in the body. Since the decomposition rate of the polymer is proportional to the number of dicarboxyl linkers included, hydrolysis rate and the size of the hydrolyzed product can be controlled by adjusting the size and number of each block.

The microspheres prepared using the multiblock polymer of the present invention can be used as drug carriers by including drugs, and any applicable drug can be used, for example, nonionic and ionic drugs. Applicable Preference is given to peptides or proteins which are preferably ionic and in particular have a large number of carboxyl and amino ionic groups in the molecule.

Examples of the peptide or protein include growth hormone (GH), interferon (IFN), leukocyte proliferation factor (G-CSF), erythrocyte producing protein (EPO), interleukin (IL), follicle stimulating hormone (FSH), nerve growth factor ( NGF), octreotide, insulin, calcitonin, tissue necrosis factor (TNF), angiogenesis factor (VEGF), epithelial growth factor (EGF), platelet growth factor (PDGF), bone formation factor (BMP), thrombolytics ( TPA), oligonucleotide, etc. are mentioned. The protein may be in natural form or modified in a polymer such as PEG. Therefore, the protein may delay the drug release from the gel when forming the metal ion complex such as a polymer and prior ^{Zn 2 +, Ca 2 +,} Cu 2 +, Mg 2 ⁺ to a mixture of the present invention.

The microsphere of the present invention may be one containing 40 to 99.9% by weight of the multiblock polymer, 0.01 to 60% by weight of the drug. The multiblock polymer solution may be used as a drug carrier as long as it exhibits phase transition and may be used at a concentration of 0.5 to 50%.

In order to prepare a peptide or protein drug delivery formulation, it is necessary to make the multiblock polymer of the present invention in an aqueous state. The polymer of the present invention does not melt well at room temperature such as 25 ° C, but shows high solubility at low temperature such as 4 ° C. Therefore, it is preferable to dissolve the polymer at low temperature. Although the amount of the multiblock polymer that can be dissolved is limited depending on the molecular weight, in the case of a multiblock polymer having a molecular weight of about 100,000, up to 30%, preferably 4 to 20% can be dissolved in water. Therefore, when the peptide, protein or water-soluble drug is mixed with the multiblock polymer solution at low temperature and avoided orally orally, the aqueous solution turns into a gel state at body temperature, thereby slowly releasing the peptide, protein drug or water-soluble drug.

Conventional microspheres are prepared by dissolving polylactic glycolic acid (PLGA) or polylactic acid (PLA) in an organic solvent and dropping them in an aqueous solution to form spherical droplets and removing the organic solvent. Instead of using organic solvents, the present invention dissolves and sprays biocompatible polymers in aqueous solvents to form droplets and uses liquefied gases, which are not harmful to the human body. It can also increase the stability.

1 is a schematic diagram showing an apparatus for manufacturing microspheres by a direct injection method.
2 is a schematic view showing an apparatus for manufacturing microspheres by an indirect injection method.
3 is a SEM photograph of the prepared microspheres.
4 is a 1 H-NMR spectrum of poloxamer disuccinate.
* Description of the major signs in the drawings
2: reaction vessel
20: vacuum pump 22: negative pressure tube
24: exhaust port 26: exhaust port
28: slope 30: valve
32: stirring blade 34: handle
4: injection part
40: nozzle 42: pressure regulating pump
6: temperature controller
60: vessel side heat exchange means 62: nozzle side heat exchange means
64: thermostat
G1: polymer solution
G2: globular (freezing particle)

Hereinafter, the present invention will be described in more detail with reference to examples in order to more clearly describe the present invention and the effects of the present invention, but these are only for explaining the present invention, and the scope of the present invention is defined by the following examples. It is not limited in any way.

EXAMPLE  One: Succinyl Dichloride  Multiblock Polymer Synthesis as a Link

10 g of Fluornic F127 (BASF; Poloxamer 407) was placed in a 100 ml 1-neck round bottom flask with a magnetic rod, and the water contained in the polymer was heated and decompressed (less than 1 torr) in an oil bath heated to 120 ° C. Was removed for 2 hours. The pressure was released and the reaction temperature was set to 100 ° C. while flowing nitrogen, then 100 ml of acetonitrile were added to the flask. A dean stark and a cooler were installed in the reaction flask. Distilled acetonitrile was removed 20 ml through dean stark to completely remove the water in the reaction, and then reacted for 24 hours by adding 96ul (1 equivalent of polymer) of succinyl dichloride to the reservoir of the dean stark apparatus. After 24 hours, 96ul of succinyl chloride was added to the reservoir of the dean stark apparatus in order to replace the terminal group of the synthesized poloxamer-407 oligomer. The synthesized poloxamer-407 oligomer was precipitated in 1 L of diethyl ether and filtered to obtain the product (8.2 g).

The product (8 g) thus obtained was again dissolved in 16 ml of methanol, precipitated and filtered through diethyl ether, and purified twice. Vacuum drying was carried out to obtain a narrow molecular weight distribution of fluorosomer oligomer (5.7 g).

The molecular weight of the floxamer oligomer was determined by GPC, the weight average molecular weight was 90,700, and the synthesis was confirmed by 1 H-NMR. The obtained NMR spectrum is shown in FIG. 4.

EXAMPLE  2: Oxalyl  Chloride as link Phloxamer Oligomer  synthesis

The same method as in Preparation Example 1 was performed except that oxalyl chloride was used as the dicarboxylic acid link. The molecular weight of the obtained fluorosomer oligomer was 91,300.

EXAMPLE  3: Adifoil Dichloride  Used as link Phloxamer Oligomer  synthesis

The same method as in Preparation Example 1 was performed except that adipoyl dichloride was used as the dicarboxylic acid link. The molecular weight of the obtained poloxamer oligomer was 96,300.

EXAMPLE  4: Suberoyl Dichloride  Used as link Phloxamer Oligomer  synthesis

The same method as in Preparation Example 1 was carried out except that suveroyl dichloride was used as the dicarboxylic acid link. The molecular weight of the obtained poloxamer oligomer was 97,800.

EXAMPLE  5: Seba coil Dichloride  Used as link Phloxamer Oligomer  synthesis

The same method as in Preparation Example 1 was performed except that sebacoyl dichloride was used as the dicarboxylic acid link. The molecular weight of the obtained poloxamer oligomer was 124,000.

EXAMPLE  6: Dodecane Oil Dichloride  Used as link Phloxamer Oligomer synthesis

The same method as in Preparation Example 1 was conducted except that dodecanedioyl dichloride was used as the dicarboxylic acid link. The molecular weight of the obtained fluorosomer oligomer was 104,000.

* EXAMPLE 7: Terephthaloyl Flock sameo using the dichloride as a link Oligomer synthesis

The same method as in Preparation Example 1 was performed except that terephthaloyl dichloride was used as the dicarboxylic acid link. The molecular weight of the obtained poloxamer oligomer was 87,000.

EXAMPLE  8: using fumaric acid as link Phloxamer Oligomer  synthesis

10 g of fumaric acid and 22 g of oxalyl chloride (double equivalent of fumaric acid) were quantified in 50 ml of acetonitrile and reacted at 50 ° C. for 6 hours. After the reaction, excess oxalyl chloride was removed in vacuo to give reactive fumaroyl dichloride. Thus synthesized fumaroyl chloride was used as the dicarboxylic acid link was carried out in the same manner as in Preparation Example 1. The molecular weight of the obtained pulloxamer oligomer was 85,400.

EXAMPLE  9: Maleic acid  Used as link Phloxamer Oligomer  synthesis

The same procedure as in Preparation Example 8 was conducted except that maleic acid was used. The molecular weight of the obtained poloxamer oligomer was 82,700.

EXAMPLE  10: Malic acid  Used as link Phloxamer Oligomer  synthesis

The same procedure as in Preparation Example 8 was carried out except for using malic acid. The molecular weight of the obtained fluorosomer oligomer was 84,000.

EXAMPLE  11: using multiblock polymer solution Microsphere  Produce

700 mg of the multiblock polymer (molecular weight 101,500) of Preparation Example 1 was dissolved in 9.3 g of distilled water and left at 4 ° C. for 2 hours to form a 7% multiblock polymer solution. The solution was sprayed through a 0.7 mm nozzle while maintaining the solution at 10 ° C. to contact liquid nitrogen, and then the vessel was kept at −20 ° C. and dried for 6 hours using a vacuum pump to obtain a microsphere.

After the final drying, the results observed using an electron microscope are shown in FIG. 3. As can be seen in FIG. 3, microspheres having a size of 10 μm to 200 μm were obtained.

EXAMPLE 12: Preparation of Microspheres Using a Multiblock Polymer Solution Mixed with Interferon

1.28 mg of interferon-alpha (Sigma) was dissolved in 10 ml distilled water, and 1.46 ml of zinc acetate (1 mg / ml) was mixed to form a molar ratio of Zn and interferon of 100: 1 to form an interferon-Zn complex. 840 mg of the multiblock polymer of Preparation Example 1 was added to this solution and stored at 4 ° C. for about 3 hours to prepare a 7% interferon-multiblock polymer solution. When the microspheres were prepared in the same manner as in Example 1, an interferon-multiblock polymer microsphere having a size of 10 μm to 200 μm was prepared.

EXAMPLE 13: A microsphere mixed with hyaluronic acid in a multiblock polymer solution Produce

700 mg of the multiblock polymer of Preparation Example 1 was placed in 9.3 ml of distilled water, and stored at 4 ° C. for 3 hours to prepare a multiblock polymer solution, and 70 mg of hyaluronic acid (Rip Co., Ltd.) was dissolved therein. An aqueous solution of mixed nolonic acid was prepared. This solution was prepared in a microsphere in the same manner as in Example 1.

EXAMPLE 14: Preparation of Mycross Peer Using Multiblock Polymer Solution Mixed with Human Growth Hormone

2.07mg of human growth hormone was dissolved in 1.66ml distilled water, and then 300mg of the multiblock polymer of Preparation Example 1 was added to this solution and stored at 4 ° C. for about 3 hours to prepare an interferon-multiblock polymer solution. Microspheres were prepared in the same manner as in Example 1 to obtain interferon-multiblock polymer microspheres having a size of 10 μm to 200 μm.

Claims (12)

Spraying a high molecular weight biocompatible water-soluble aqueous polymer solution having a weight average molecular weight of 25,000 to 1,000,000 daltons through a nozzle at a pressure in the range of 0.5 to 15 bar to form a droplet;
The formed droplets are contacted with the liquefied gas and frozen to form frozen particles;
Removing the liquefied gas and freeze drying the frozen particles at a temperature in the range of −70 ° C. to −1 ° C. and a pressure of 1 Torr or less,
The biocompatible water-soluble polymer is hyaluronic acid (Hyaluronic acid), dextran, gelatin, collagen, chitosan, methyl cellulose (MC), ethyl cellulose (EC), hydroxyethyl cellulose (HEC), methyl hydroxyethyl cellulose (MHEC), hydroxymethyl cellulose, hydroxypropyl methyl cellulose (HPMC), and at least one selected from the group consisting of hydroxypropyl cellulose,
Method for producing a polymeric microsphere. The method of claim 1, wherein the injection is performed by a direct injection method or an indirect injection method. The method of claim 1, wherein the liquefied gas is selected from the group consisting of liquefied nitrogen, liquefied oxygen, and liquefied helium. The method of claim 1, further comprising the step of agitating in contacting the droplets with a liquefied gas to prevent tangling of the droplets. The method of claim 1, wherein the biocompatible polymer aqueous solution contains the biocompatible polymer at a concentration of 3 to 25 wt% in an aqueous solvent selected from the group consisting of distilled water, acetate buffer solution and phosphate buffer solution. According to claim 1, wherein the biocompatible polymer aqueous solution is growth hormone (GH), interferon (IFN), leukocyte proliferation factor (G-CSF), erythropoietin (EPO), interleukin (IL), follicle stimulating hormone (FSH ), Nerve growth factor (NGF), octreotide, insulin, calcitonin, tissue necrosis factor (TNF), angiogenesis factor (VEGF), epithelial growth factor (EGF), platelet growth factor (PDGF), bone formation factor ( BMP), thrombolytics (TPA) and oligonucleotides further comprises at least one bioactive material selected from the group consisting of. According to claim 1, wherein the biocompatible polymer aqueous solution is stabilizer selected from the group consisting of PBS, amino acids, carbohydrates, fatty acids and surfactants; And at least one selected from the group consisting of excipients selected from the group consisting of lactose, dextran, mannitol and sucrose in an amount of 1% to 20% by weight based on the weight of the polymer. The method of claim 1, wherein the spraying is performed while maintaining the temperature of the polymer solution at 5 to 12 ℃. Hyaluronic acid, dextran, gelatin, collagen, chitosan, methylcellulose (MC), ethylcellulose (EC), hydroxyethylcellulose (HEC), methylhydroxyethylcellulose (MHEC), hydroxy At least one polymer selected from the group consisting of methyl cellulose, hydroxypropyl methyl cellulose (HPMC), and hydroxypropyl cellulose, for preparing microspheres from a high molecular weight biocompatible aqueous solution having a weight average molecular weight of 25,000 to 1,000,000 daltons As a manufacturing apparatus,
A reaction vessel in communication with the vacuum pump through the negative pressure pipe, the inside of which is formed as a vacuum, and the gas generated therein can be exhausted through the exhaust port, and an inclined surface inclined around the outlet port is formed on the inner bottom surface;
The nozzle includes a nozzle extending inwardly through the reaction vessel, the nozzle is in communication with the pressure control pump to spray the polymer solution supplied at a constant pressure into the reaction vessel to form a droplet of uniform shape Injection unit to be;
A container side heat exchange means covering a portion of the outer surface of the reaction vessel 2, and nozzle side heat exchange means surrounding a portion exposed from the nozzle to the outside of the reaction vessel, and these heat exchange means are provided by a temperature controller. A temperature control unit which circulates the pressure-exchanging heat exchange medium to maintain the inner atmosphere of the reaction vessel and the vicinity of the nozzle at a predetermined temperature or less so that the polymer solution is not gelled; Microsphere manufacturing apparatus equipped with. The microsphere manufacturing apparatus according to claim 9, wherein the injecting part is configured to be disposed above or to the side of the reaction vessel. 10. The method of claim 9,
The reaction vessel is a microsphere manufacturing apparatus, characterized in that the configuration with a stirring blade inside. A microsphere prepared by the method according to any one of claims 1 to 8, having a particle size of 10 µm to 500 µm and a porous surface.

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