KR101741982B1 - Porous biodegradable polymer microsphere for dual-loaded drug delivery and method for producing thereof - Google Patents

Abstract

Porous biodegradable polymeric microparticles for drug delivery and a process for producing the same. The porous biodegradable polymeric microparticles and the method for producing the porous biodegradable polymeric microparticles and the method for producing the same are capable of simultaneously releasing the two drugs in a predetermined amount for a long period of time at the same time and preventing damage or degeneration of the drug by chemical bonding or organic solvent And the amount of desired drug can be efficiently carried.

Description

TECHNICAL FIELD [0001] The present invention relates to porous biodegradable polymer microparticles for drug delivery, and biodegradable polymer microspheres for dual-

The present invention relates to porous biodegradable polymeric microparticles for drug delivery and a process for producing the same.

It is important to design the dosage form so that the drug delivery microparticles can efficiently deliver the drug for treatment of the disease to the treatment site, thereby reducing side effects of the drug, increasing the patient compliance with the drug, and maximizing the efficacy and effectiveness of the drug. Particularly, the drug-transferring fine particles using the biodegradable polymer should be able to easily incorporate a lipid-soluble or water-soluble physiologically active substance into the microparticles, and be capable of supporting the drug in the body for a predetermined period of time, And should have persistence to release sufficient drug during the desired period of time after reaching the target point without releasing the drug initially in the human body.

Examples of the method for producing fine particles using the biodegradable polymer include a solvent evaporation drying method (U.S. Patent No. 4,652,441), a phase separation method (U.S. Patent No. 4,675,189), a spray drying method (U.S. Patent No. 6,709,650) Patent No. 5,019,400) and the like are known. As a method of producing the porous fine particles, a salt leaching / gas foaming method and a phase separation method using a solvent / non-solvent are known. The salt leaching / gas foaming method uses ammonium bicarbonate (NH4HCO3) or the like as an effervescent material (Kim et al., Biomaterials 27: 152-159, 2006). In the solvent / non-solvent separation method, the polymer is miscible with the solvent used after the polymer is dissolved in the solvent, (Por et al., Polym. Adv. Technol. 16: 622-627, 2005).

However, the microparticles thus far developed have a high initial release rate and have a disadvantage of having a delay time and a low drug duration after a certain amount is released. In addition, it is difficult to control the size of fine particles using W / O / W (water in oil in water) dual emulsion, and the amount of the drug to be supported is limited by the internal surface area. Surface inclusion of growth factors such as internal inclusion, surface modification through chemical treatment, change in organic solvent or pH during ionic bonding, and the like, leading to damage of drugs such as aggregation, unfolding or denaturation of growth factors.

One aspect of the invention is a porous biodegradable polymeric microparticle hollow hollow, wherein the microparticle has a first pharmaceutically active substance carried on an outer surface having pores, and a second pharmaceutically active substance in an inner void space Wherein the hydrogel is supported on the carrier.

In another aspect, there is provided a method for preparing a biodegradable polymer, comprising: dissolving a biodegradable polymer and a first pharmaceutically active substance in an organic solvent to prepare a polymer solution containing the first pharmaceutically active substance; Adding a pore forming agent to the polymer solution to prepare a W / O (water in oil) emulsion; Adding the W / O emulsion to an emulsion stabilizer solution to prepare a water in oil in water (W / O / W) emulsion; And immersing the W / O / W emulsion in a hydrogel containing a second pharmaceutically active substance. The present invention also provides a method for preparing porous hollow biodegradable polymeric microparticles.

One aspect of the invention is a porous biodegradable polymeric microparticle hollow hollow, wherein the microparticle has a first pharmaceutically active substance carried on an outer surface having pores, and a second pharmaceutically active substance in an inner void space Wherein the hydrogel is supported on the support.

The term "microsphere" may refer to a particle for encapsulating a drug in a formulation and is used interchangeably with microparticles, nanoparticles or nanospheres. Particulates may be in the micron or micrometer range or smaller in size, for example, from about 0.01 microns to about 1000 microns, from about 1 microns to about 500 microns, from about 300 microns to about 500 microns, or from about 400 microns to about 500 microns Lt; RTI ID = 0.0 > um. ≪ / RTI > The particulate may be of any shape, for example, an approximate spherical shape (e.g., microspheres), spherical, elliptical, cylindrical,

Further, the fine particles may be porous and hollow. That is, the outer surface may be porous with pores and have an empty space therein. The diameter of the inner empty space may be, for example, from about 200 to about 400 microns, from about 200 to about 300 microns, from about 300 to about 400 microns, and from about 350 to about 400 microns. The pore diameter may also be, for example, from about 2 to about 80 microns, from about 5 to about 40 microns, and from about 5 to about 20 microns.

In the present invention, the term "pharmacologically active agent" is a substance that enhances or inhibits the function of a living body when living organisms are living. The term " pharmacologically active agent " Which can be used to correct an abnormal condition. The pharmacologically active substance may be, for example, a growth factor, a growth hormone, a peptide or protein drug, an antiinflammatory agent, an anticancer agent, an antiviral agent, a sex hormone, an antibiotic, an antimicrobial agent or a low molecular compound. More specifically, dexamethasone, Progesterone haloperidol, thiothixene, olanzapine, clozapine, bromperidol, pimozide, risperidone, ziprasidone, Diazepma, ethyl loflazepate, alprazolam, nemonapride, fluoxetine, sertraline, venlafaxine, < RTI ID = 0.0 & But are not limited to, donepezil, tacrine, galantamine, rivastigmine, selegiline, ropinirole, pergolide, trihexepenidyl ( for example, trihexyphenidyl, bromocriptine, benztropine, colchicine, nordazepam, etizolam, bromazepam, clotiazepam, But are not limited to, mexazolum, buspirone, goserelin acetate, somatotropin, leuprolide acetate, octreotide, cetrorelix Gonadotropin, fluconazole, itraconazole, mizoribine, cyclosporin, tacrolimus, naloxone, and the like) Naltrexone, cladribine, chlorambucil, tretinoin, carmusitne, anagrelide, doxorubicin, anastrozole, , Idarubicin Cisplatin, dactinomycin, docetaxel, paclitaxel, raltitrexed, epirubicin, letrozole, mefloquine, prime queen, or at least one selected from the group consisting of primaquine, oxybutynin, toltrerodine, allylestrenol, lovostatin, simvastatin, provastatin, atrovastatin, alendronate, but are not limited to, alendronate, salcatonin, raloxifene, oxadrolone, conjugated estrogen, estradiol, estradiol valerate, estradiol benzoate, May be estradiol benzoate, ethinylestradiol, etonogestrel, levonorgestrel, tibolone, norethisterone, and the like. And may be selected from the group consisting of interleukin, interferon, tumor necrosis fiactor, insulin, glucagon, growth hormone, gonadotropin, oxytocin (hormone contraction hormone , oxytocin), thyroid stimulating hormone, parathyroid hormone, calcitonin, colony stimulation factor, erythropoietin, thrombopoietion, An insulin-line growth factor, an epidermal growth factor, a platelet-derived growth factor, a transforming growth factor, a fibroblast growth factor fibroblast growth factor, vascular endothelial growth factor or bone morphogenetic protein (BMP). The bone morphogenetic protein may be selected from the group consisting of BMP-2, BMP-3, BMP-3b, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8, BMP- 12, BMP-13, BMP-14, BMP-15, BMP-16, BMP-17, BMP-18 and combinations thereof. The first pharmaceutically active composition and the second pharmaceutically active composition may or may not be the same and the first pharmaceutically active composition and the second pharmaceutically active composition may exhibit a synergistic effect on a particular pathological condition But it is not limited thereto.

The term "biodegradable polymer" (used interchangeably with "biocompatible" or "bioabsorbable") can mean any material that can be absorbed in vivo. The biodegradable material does not need to be chemically degraded or chemically degraded in vivo, but may be dispersed or dissolved in vivo in such a manner as to release a therapeutically effective amount of the drug for a significantly longer period of time than the normal in vivo lifetime of the drug Or < / RTI > The biodegradable material can be any non-toxic polymer that can be degraded in vivo. Examples include polylactic acid, polylactide, polylactic-co-glycolic acid, polylactide-co-glycolide (PLGA) Selected from the group consisting of polyglycolic acid, polyglycolic acid, polyglycerol, polyglycerol, polyglycerol, polyglycerol, polyglycerol, polyglycerol, polyglycerol, . The biodegradable material may be a homopolymer or copolymer, or any combination or blend of polymers. Examples of biodegradable polymeric materials may include polymers consisting of monomers such as esters, ethers, anhydrides, amides, or orthoesters that produce physiologically acceptable degradation products upon degradation. The polymeric material may be cross-linked or non-cross-linked. When crosslinked, they can be crosslinked to a weakly, e.g., 5% or less than 1%.

In addition, the second pharmaceutically active material may be one contained in a hydrogel, i.e., one present on a hydrogel matrix. The hydrogel may be cross-linked or non-cross-linked, and cross-linked may be cross-linked to less than about 40%.

The term "hydrogel" may refer to a three-dimensional network formed by crosslinking a hydrophilic polymer with a covalent or noncovalent bond. Due to the hydrophilicity of the constituent material, it absorbs a large amount of water in an aqueous solution and an aqueous environment and swells but does not dissolve by the crosslinking structure. Therefore, a hydrogel having various shapes and properties can be produced depending on the constitution and the manufacturing method, and it may be one having an intermediate property between a liquid and a solid because it generally contains a large amount of water.

The preparation method of the hydrogel can be prepared by a known method. For example, hydrogels can be prepared by ionic cross-linking, covalent cross-linking, photo cross-linking, or cell cross-linking. Ionic crosslinking can be produced by mixing an aqueous solution of a polymer and an aqueous solution of a divalent cation. The divalent cation can be, for example, Ca < ^{2 + & gt ;} , and can include calcium chloride, calcium sulfate, calcium carbonate and the like. Crosslinking using covalent bonds can form hydrogels having higher physical properties than crosslinking using ionic bonds. For example, PEG-diamines of various molecular weights can be used as crosslinking agents. Photocrosslinking is a method that can form a gel in situ and can be exposed to ultraviolet light at a specific wavelength to produce a hydrogel. Cell crosslinking is a crosslinking method in which the cell itself acts as a crosslinking agent. For example, a hydrogel can be formed by introducing an arginine-glycine-aspartic acid (RGD) peptide into a polymer main chain to induce specific interactions.

The hydrogel may be, for example, a natural hydrogel or a synthetic hydrogel, and more particularly, a calcium alginate, an alginic acid, a hyaluronic acid, a silk, a keratin, a collagen, gelatin, elaster, cellulose, or chitosa hydrogels.

The polymer biodegradable fine particles may be prepared by a known method. For example, solvent evaporation / extraction or solvent decomposition method through an emulsion, spray drying method or phase separation method, O / W (oil-in-water) type including polymer compound, drug and dispersion solvent, O An oil-in-oil type oil or an oil-in-water (W / O / W) oil type emulsion is prepared and the oil is coagulated with fine particles, a polymer compound, A method in which air is sprayed into the air to solidify the polymer to coagulate it into fine particles, or a method in which a non-solvent is added to an organic solvent including a polymer compound or a drug to induce phase separation and then transferred to an additional solvent to solidify the polymer, The polymer fine particles can be produced.

Among the methods for producing polymeric fine particles, in order to prepare an emulsion by agglomeration with polymer fine particles, an O / W type, an O / O type or a W / O / W type emulsion containing a polymer compound, a drug and a dispersion solvent . Conventional methods known in the art can be used for the production of the emulsion, and more specifically, for the production of O / W type or O / O type emulsion, a dispersion phase containing a polymer compound and a drug is added to a dispersion solvent In order to prepare a W / O / W type emulsion, a W / O type emulsion is prepared by emulsifying an aqueous solution in which a drug is dissolved in a solvent in which a polymer compound is dissolved, W emulsion can be produced.

Another aspect provides a drug delivery composition comprising the microparticles.

The drug delivery vehicle may comprise a pharmaceutically acceptable carrier and a pH adjusting agent. The drug delivery vehicle may be in the form of an injection, and since the particulates are very small, they can be delivered through very small needles, such as about 25 gauge to 30 gauge. The use of small needles can reduce the risk of serious side effects from needles.

Another aspect provides a method for producing porous biodegradable polymeric microparticles hollowed out.

The method comprises: dissolving a biodegradable polymer and a first pharmaceutically active substance in an organic solvent to prepare a polymer solution containing a first pharmaceutically active substance to prepare a dispersed phase; A pore forming agent is added to the dispersed phase to form an oil-in-water type, an oil-in-oil type or a water in oil-in-water (W / O / W) ) ≪ / RTI > Wherein the oil-in-water, O / O or W / O / W oil-in-water emulsion comprises a second pharmaceutically active substance, And immersing it in a hydrogel.

The method may further comprise the steps of: dissolving the biodegradable polymer and the first pharmacologically active substance in an organic solvent to prepare a polymer solution containing the first pharmacologically active substance; Adding a pore forming agent to the polymer solution to prepare a W / O (water in oil) emulsion; Adding the W / O emulsion to an emulsion stabilizer solution to prepare a water in oil in water (W / O / W) emulsion; And immersing the W / O / W emulsion in a hydrogel comprising a second pharmaceutically active material.

The biodegradable polymer, the pharmaceutically active substance, and the fine particles are as described above.

The "dispersed phase" of the present invention may mean that the polymer compound and the drug are dissolved in an organic solvent and mixed

The method for preparing porous hollow biodegradable polymeric microparticles may include the step of dissolving the biodegradable polymer and the first pharmaceutically active substance in an organic solvent to prepare a polymer solution containing the first pharmaceutically active substance . The biodegradable polymer may be added in an amount of 1 to 20 wt% based on the organic solvent, and the first pharmaceutically active material may be added in an amount of 0.1 to 10 wt% based on the biodegradable polymer. The organic solvent of step (1) may vary depending on the kind of the polymer used, but may include methylene chloride, chloroform, carbon tetrachloride, acetone, dioxane, tetrahydrofuran, or hexafluoro isopropanol.

The hollow biodegradable porous polymer microparticles may be prepared by adding a pore-forming agent to the polymer solution to prepare a W / O emulsion. The added Sucrose Solution may contain about 0.3 g / ml to about 3 g / ml, such as 0.5 to 1.5 g / ml, 0.8 to 1.5 g, of sucrose and DW (distilled water) / ml, or 1.2 g / ml (7.2 g / 6 ml). The pore former may be, for example, sucrose. The step of preparing the W / O emulsion may be prepared by adding the pore-forming agent, followed by stirring at about 100 to 2,500 rpm for 10 seconds to 5 minutes.

The method for producing the microporous porous biodegradable polymer microparticles may include adding the W / O emulsion to the emulsion stabilizer solution to prepare a W / O / W emulsion. The emulsion stabilizer may be added at a concentration of 0.3% (w / v) to 5.0% (w / v) to facilitate the formation of particles. Examples of the emulsion stabilizer include an aqueous dispersion solvent or a non-aqueous dispersion solvent, and an aqueous dispersion solvent is used for the O / W type and W / O / W type emulsion, and a non- Can be used. Examples of the aqueous dispersion solvent include hydrophilic emulsifiers such as polyvinyl alcohol, poloxamer, polyethylene glycol and Polysorbate series (for example, Polysorbate 20, Polysorbate 60, Polysorbate 65, Polysorbate 80, Polysorbate 85), or a co-solvent thereof may be used. As the non-aqueous dispersion solvent, there can be used a lipophilic emulsifier, for example, a silicone oil containing an emulsifier such as Glycerin Esters of Fatty Acids, lecithin, vegetable oil, toluene or xylene.

The step of preparing the W / O / W emulsion may further include, after adding the emulsion stabilizer, evaporating the organic solvent and separating the fine particles by centrifugation.

The method for preparing porous hollow biodegradable polymeric microparticles may comprise immersing the W / O / W emulsion in a solution containing a second pharmaceutically active material. The step of supporting the second pharmacologically active substance may include carrying the second pharmacologically active substance in a hydrogel. The hydrogels are as described above. Also, the concentration of the hydrogel may be from about 10 mg / ml to about 500 mg / ml, from about 10 mg / ml to about 400 mg / ml, from about 10 mg / ml to about 350 mg / ml, About 300 mg / ml, or about 20 mg / ml to about 250 mg / ml. By immersing the W / O / W emulsion in the second pharmaceutically active substance / hydrogel mixture, the porous biodegradable polymer microparticles can be produced.

In addition, the step may further include a step of performing ultrasonic treatment or decompression after immersion, and centrifuging at about 1,000 rpm to 30,000 rpm for about 1 minute to 1 hour to separate the fine particles. In addition, after the fine particles are separated by centrifugation, a drying process such as freeze-drying may be carried out to produce fine particles.

According to the porous biodegradable polymer microparticles according to one embodiment, the initial release is not large by controlling the concentration of the hydrogel without the chemical treatment, and the two drugs can be simultaneously released for a long period of time in a constant amount.

According to one embodiment of the method for producing porous biodegradable polymeric microparticles, it is possible to prevent damage or denaturation of a drug due to chemical bonding or organic solvent, and to efficiently support a desired amount of drug.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a scanning electron micrograph of a porous biodegradable microparticle according to an embodiment of the present invention, according to the amount of sucrose; (d, i) 7.2 g, (e, j) 14 g, above: surface morphology, below: section.
2 is a scanning electron micrograph of a porous biodegradable microparticle according to an embodiment of the present invention in accordance with the amount of dexamethasone; 5 mg, (c, g, k) 10 mg, (d, h, 1) 20 mg.
FIG. 3 is a scanning electron micrograph of a porous biodegradable microparticle according to an embodiment of the present invention, according to the amount of collagen; FIG. (a, e) 0.025%; (b, f) 0.5%; (c, g) 10%; (d, h) 20%
FIG. 4 is a graph showing the size distribution and porosity according to the amount of sucrose in the porous biodegradable microparticles according to one embodiment.
FIG. 5 is a graph showing the size distribution and porosity of the porous biodegradable microparticles according to one embodiment of the present invention in accordance with polyvinyl alcohol. FIG.
FIG. 6 is a graph showing a water absorption rate according to the amount of sucrose in the porous biodegradable microparticles according to one embodiment.
FIG. 7 is a graph showing FT-IR test results according to dexamethasone concentration of porous biodegradable fine particles according to one embodiment.
8 is a graph showing the dexamethasone and BMP2 release profiles according to the collagen concentration of the porous biodegradable microparticles according to one embodiment.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: Preparation of porous biodegradable microparticles for dual drug delivery

1. dexamethasone Supported  Preparation of Porous Biodegradable Polymer Fine Particles

A water-in-oil-in-water (W / O / W) solvent evaporation technique was used to prepare porous biodegradable polymeric microspheres.

Specifically, different amounts of sucrose (0, 1.8, 3.6, 10, 10, 10, 20, 30, 40, 50, 7.2, 14g) was added. Then, a water in oil (W / O) emulsion was prepared using a Powergen 700 homogenizer and homogenized at 1,500 rpm for about 3 minutes. To the above water-in-oil type emulsion was added 300 ml of a solution of 0.6% (w / v), 1.25% (w / v) and 2.5% (w / v) polyvinyl alcohol (PVA) as an emulsion stabilizer. Thereafter, the emulsion was again emulsified with stirring at 200 rpm for 4 hours to prepare a W / O / W (water in oil in water) emulsion. The methylene chloride of the prepared water-in-oil type emulsion was evaporated and separated by centrifugation. Thereafter, the cells were washed with distilled water at 200 rpm for 6 hours, and then lyophilized to prepare porous biodegradable polymer microparticles having dexamethasone and finally hollow.

2. Dexamethasone and BMP2 end Supported  Preparation of Porous Biodegradable Polymer Fine Particles

BMP2 was additionally supported on the microparticles prepared in Example 1 (1) to prepare dual drug delivery vehicles.

Specifically, collagen type I solution (0.025 g / ml, 0.05 g / ml, 0.1 g / ml, 0.2 g / ml) was stirred at 37 ° C at 2000 rpm for 1 hour, Was added thereto to prepare a collagen hydrogel containing BMP-2. To encapsulate the collagen hydrogel containing the BMP-2 in the microparticles prepared in Example 1, 500 mg of the microparticles were immersed in 5 ml of the collagen / BMP2 mixture and sonicated for 5 minutes. The dexamethasone and BMP2-loaded microparticles were then separated by centrifugation (16,000 rpm, 20 minutes) and washed 3 times with deionized water to remove excess collagen / BMP2 mixture. Thereafter, the cells were lyophilized for 2 days to finally produce dexamethasone and BMP2-loaded porous biodegradable polymer microparticles.

Example  2: Characterization of porous biodegradable microparticles for dual drug delivery

1. Morphology analysis

The morphology of the fine particles prepared in Examples 1 and 2 according to the amounts of dexamethasone and sucrose was analyzed.

Specifically, each sample was frozen in liquid nitrogen and sectioned into surgical blades. Thereafter, gold / palladium was thin-coated, mounted on an aluminum stub using carbon tape, observed with a scanning electron microscope (SEM; Jeol 6400-LINK AN 10000 at 10 KV) Respectively.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a scanning electron micrograph of a porous biodegradable microparticle according to an embodiment of the present invention, according to the amount of sucrose; (d, i) 7.2 g, (e, j) 14 g, above: surface morphology, below: section. As shown in Fig. 1, porous biodegradable microspheres hollow in the microspheres added with a certain amount of sucrose are formed.

2 is a scanning electron micrograph of a porous biodegradable microparticle according to an embodiment of the present invention in accordance with the amount of dexamethasone; 5 mg, (c, g, k) 10 mg, (d, h, 1) 20 mg. As shown in FIG. 2, even when 5 mg of dexamethasone is introduced, it can be confirmed that there is no significant difference in morphology with the microparticles into which dexamethasone is not introduced.

FIG. 3 is a scanning electron micrograph of a porous biodegradable microparticle according to an embodiment of the present invention, according to the amount of collagen; FIG. (a, e) 0.025%; (b, f) 0.5%; (c, g) 10%; (d, h) 20%. As shown in Fig. 3, it can be confirmed that porous biodegradable fine particles carrying collagen hydrogel containing dexamethasone and BMP2 are formed.

2. Size dispersion and porosity analysis

The size dispersion and porosity of the microspheres prepared in Examples 1 and 2 according to the amounts of sucrose and polyvinyl alcohol were analyzed.

Specifically, the microparticles were suspended in water and images were obtained with an optical microscope (Leica DM / Leitz DM RBE Microscope system, Leica Microsystems, Wetzlar, Germany). The size distribution and porosity were analyzed for the image using image software (image J). The results of the analysis are shown in FIGS. 4 and 5.

FIG. 4 is a graph showing the size distribution and porosity according to the amount of sucrose in the porous biodegradable microparticles according to one embodiment. As shown in FIG. 4, it can be confirmed that the porosity can be controlled by the fine particle manufacturing method according to one embodiment. When the concentration of sucrose is 7.2 g / 6 ml, the diameter is 400 to 500 μm, the porosity is 80% Or more.

FIG. 5 is a graph showing the size distribution and porosity of the porous biodegradable microparticles according to one embodiment of the present invention in accordance with polyvinyl alcohol. FIG. As shown in FIG. 5, it can be confirmed that the particle size of the fine particles, that is, the porosity can be controlled by controlling the concentration of the PVA solution by the fine particle production method according to one embodiment.

3. Analysis of moisture uptake and supporting efficiency

The water uptake and loading efficiency according to the amount of sucrose in the fine particles prepared in Examples 1 and 2 were analyzed.

Specifically, after the weight of the dried fine particles was measured, it was immersed in 70% ethanol. After three washes with demineralized water, the cells were incubated in a phosphate buffer solution for 12 hours. After wiping excess moisture on the surface of the microparticles, the weight of the wet microparticles was measured. The water absorption rate was calculated by the following equation, and the results are shown in FIG.

Wherein Ww is the weight of the wet microparticles and Wd is the weight of the dry microparticles.

FIG. 6 is a graph showing a water absorption rate according to the amount of sucrose in the porous biodegradable microparticles according to one embodiment. As shown in FIG. 6, the porous biodegradable microparticles according to one embodiment showed the highest water uptake rate in a sample to which 7.2 g of sucrose was added. As a result, it can be seen that the porous biodegradable fine particles according to one embodiment can contain a large amount of water therein, so that the internal void is large.

4. Dexamethasone and BMP2  Emission profile analysis

FT-IR (Fourier Transform Infra-Red Spectroscopy) was performed to confirm whether dexamethasone was present in the fine particles prepared in Example 1 (1). Specifically, the sample was pulverized and mixed with KBr, followed by pelleting. FTIR spectra were recorded with 48 scans (Shimadzu e IR prestige 21) at a resolution of 2 cm < 1 > and the results are shown in Fig.

In order to analyze the dexamethasone and BMP2 release profiles of the microparticles prepared in Example 1, 100 mg of porous hollow microparticles were used, and blank PLGA microparticles were used as a control. Specifically, the microparticles were placed in 1 ml PBS and placed on an orbital shaker. PBS was collected at different time intervals, frozen, supplemented with 1 ml of PBS, and the concentration of BMP2 was analyzed using ELISA KIT. Similar to the above, the microparticles were added to 2 ml of PBS (pH 7.4) to determine the release concentration of dexamethasone. The initial concentration of supported dexamethasone was quantified using a UV spectrophotometer at 235 nm and the amount of dexamethasone released was determined using an ultraviolet spectrometer at 242 nm. The loading efficiency (%) was determined as the ratio of the supported amount of dexamethasone to the initial amount, and the result is shown in FIG.

FIG. 7 is a graph showing FT-IR test results according to dexamethasone concentration of porous biodegradable fine particles according to one embodiment. As shown in FIG. 7, it was confirmed that the porous biodegradable fine particles according to one embodiment can contain a significant amount of dexamethasone.

FIG. 8 is a graph showing the release profile of dexamethasone (left side in FIG. 8) and BMP2 (right side in FIG. 8) according to the collagen concentration of the porous biodegradable fine particles according to one embodiment. As shown in FIG. 8, it was confirmed that the porous biodegradable microparticles according to one embodiment did not release the initial release of dexamethasone and BMP2 as the concentration of the collagen hydrogel increased, and were released for a long period of time in a constant amount.

Claims (15)

delete delete delete delete delete delete delete delete delete Dissolving the biodegradable polymer and the first pharmacologically active substance in an organic solvent to prepare a polymer solution containing the first pharmacologically active substance;
Adding a pore forming agent to the polymer solution to prepare a W / O (water in oil) emulsion;
Adding the W / O emulsion to an emulsion stabilizer solution to prepare a water in oil in water (W / O / W) emulsion; And
Immersing the W / O / W emulsion in a hydrogel comprising a second pharmaceutically active material,
Wherein the biodegradable polymer is polylactide-co-glycolide (PLGA), the organic solvent is methylene chloride, the pore-forming agent is sucrose, the hydrogel is a collagen hydrogel,
Wherein the pore-forming agent is added at a concentration of 0.8 to 1.5 g / ml to water. [Claim 11] The method according to claim 10, wherein the biodegradable polymer is added in an amount of 1 to 20% by weight based on the organic solvent. [Claim 11] The method according to claim 10, wherein the first pharmaceutically active substance is added in an amount of 0.1 to 10% by weight based on the biodegradable polymer. delete 11. The method of claim 10, wherein the emulsion stabilizer is added at a concentration of 0.3% (w / v) to 5.0% (w / v). 11. The method of claim 10, wherein the concentration of the hydrogel comprising the second pharmaceutically active material is from 10 mg / ml to 500 mg / ml.

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