KR101695440B1 - A Method for manufacturing a biodegradable polymeric microsphere type sustained-release formulation having comprising a fluoroquinolone antibiotics - Google Patents

Abstract

The present invention relates to a method for producing a biodegradable polymeric microparticle sustained release preparation comprising a marbofloxacin crystal powder, which is a fluorinated quinolone antibiotic, as a drug. The sustained-release preparation comprises a quinolone antibiotic including maglobulin and / or a pharmaceutically acceptable salt thereof, a biodegradable polymer, and a poloxamer-based polymer as an ionic surfactant.
The manufacturing method according to the present invention is characterized in that magofuroxacin crystal powder is composed of biodegradable polymer and poloxamer using solid-in-oil-in-water (solvent intra-extraction-evaporation) It is possible to increase the productivity by simple manufacturing process, to maximize the filling amount and sealing efficiency of the drug in the microparticle, to suppress the initial excessive release, and to release the drug continuously in the body for a certain period of time.

Description

Technical Field [0001] The present invention relates to a biodegradable polymer microsphere sustained-release formulation containing a fluorinated quinolone antibiotic,

The present invention relates to a sustained-release preparation containing a drug and a method for producing the same. More particularly, the sustained-release preparation is a drug delivery system in which a fluorinated quinolone antibiotic is loaded on a biodegradable polymeric microparticle in a high amount, maximizing the amount of drug loading and filling efficiency in the microparticles, suppressing initial overexpression, To a method for producing a sustained-release preparation capable of inducing sustained release.

Marbofloxacin is a 9-fluoro-2,3-dihydro-3-methyl-10- (4-methylpiperazin-1-yl) -7-oxo-7H-pyrido [ 3-ij] [1,2,4] benzoxadiazine-6-carboxylic acid], C ₁₇ H ₁₉ FN ₄ O ₄ , with the carbonyl group and the carboxyl group being in plane with the quinoline ring. Marbophloxacin is a third-generation fluorogenic quinolone antimicrobial and antibiotic developed for animals. It is a DNA-gyrase (an enzyme that supercoils a double helix of topoisomerase II DNA) and topoisomerase IV , Phagosomal enzymes) and exhibits an antibiotic effect. This broad spectrum antibiotics affects various Gram-negative and Gram-positive bacteria and has excellent efficacy for skin, respiratory and urinary tract infections. It is administered intravenously or intramuscularly at a dose of 2 mg of furoxacin per kg of body weight to livestock such as pigs or cattle, and the administration period is 3 to 5 days. The release period before shipment is 2 to 6 days depending on the species. Fluorinated quinolone antibiotics are bactericides and exhibit concentration-dependent bactericidal action. If the plasma concentration of the drug is increased to about 30 times the minimum inhibitory concentration (MIC), the bactericidal action becomes stronger. However, excessive use of antibiotics indefinitely or long-term use of livestock resistant bacteria appeared to reduce the effectiveness of livestock disease treatment. In addition, antibiotic resistant bacteria produced in livestock can be transmitted to humans through livestock food and animal husbandry environments, making it difficult to treat. More specifically, in the case of fluoroquinolone antibiotics, resistance to MRSA, Pseudomonas, coagulase-negative Staphylococcus aureus and E. coli appeared due to chromosomal mutation. Fluorinated quinolone antibiotics are excellent antibiotics with a wide range of antimicrobial properties and good pharmacokinetic properties. However, when the resistant bacteria are generated due to overuse of the drug, the clinical usefulness of the drug may be limited.

[Formula 1]

A sustained-release injection formulation refers to an injection formulation that is formulated so that the drug can be continuously and uniformly released while maintaining biological activity in the body during subcutaneous or intramuscular injection. Conventionally, such conventional methods of producing the sustained-release formulations are known as the coacervation method, the melt injection method, the spray drying method, and the solvent evaporation method. Among these methods, the solvent evaporation method is classified into the dual emulsion evaporation method (W / O / W emulsion) and the single emulsion evaporation method (O / W emulsion). However, such microemulsions using the multiple emulsion method have disadvantages such as high initial release rate and dissolution of the drug as an active ingredient in an organic solvent or water to form microspheres, which may cause disadvantages such as changes in physical properties of the drug, loss of stability, see.

Various attempts have been made to increase the rate of drug encapsulation of such sustained-release formulations or to produce microparticles which are easy to manufacture and have relatively low initial over-release, but are still unsatisfactory. (1983) 62-74), microspheres were prepared by double emulsion evaporation method, and then the active ingredient, vaccine antigen, was loaded through micropores of microspheres (Tg) of the biodegradable polymer to prevent the initial release by blocking the micropores. However, such a manufacturing method has a serious disadvantage that the drug can be denatured by heat. In addition, the addition of protein trapping agent significantly increased the drug loading efficiency (~97%), but the total loading of active material (1.4 ~ 1.8%) was significantly lower. This method has a disadvantage in that the physiologically active substance can be denatured by heat, and it is also disadvantageous in that it can not be applied to the encapsulation of a high dose drug. Also, it is difficult to produce by a complicated manufacturing process.

Accordingly, the present invention provides a method for maximizing the filling amount and filling efficiency of the drug in the microparticle, suppressing the initial release, minimizing the number of administrations, and minimizing the sterilization concentration of the fluoride quinolone antibiotic, The present invention provides a method for producing a biodegradable polymer sustained-release microparticle which induces sustained and uniform release of a drug.

The present invention provides a method for producing sustained-release microparticles for solving the above problems. (A) mixing a non-aqueous solvent and a cosolvent to prepare a first mixed solution, adding a biodegradable polymer and a surfactant to the first mixed solution in step (a) simultaneously or sequentially to form a second mixed solution (C) preparing an S / O suspension by adding a drug to the second mixed solution; (c) preparing an S / O / W emulsion by mixing the S / O suspension and an aqueous medium; (d) a step (e) in which the co-solvent diffuses and evaporates from the S / O / W emulsion to the aqueous medium to produce microparticles. In addition, the manufacturing method according to the present invention may further comprise a step (f) of drying the microparticles produced in the step (e).

Here, the non-aqueous solvent may have a boiling point of 25 占 폚 to 85 占 폚.

The co-solvent may be one having a boiling point lower than the boiling point of the non-aqueous solvent. In the step a), the mixing ratio of the non-aqueous solvent and the co-solvent may be 20% by volume to 50% by volume of the non-aqueous solvent and 50% by volume to 80% by volume of the co-solvent.

The surfactant in step b) may be a poloxamer-based polymer.

The surfactant in step b) may be added in an amount of more than 0 to 0.5 parts by weight, preferably 0.2 to 0.4 parts by weight based on 1 part by weight of the biodegradable polymer.

In addition, the biodegradable polymer may be selected from the group consisting of polylactide (PLA), polyglycolide (PGA), poly (lactide-co-glycolide) (PLGA), and poly (lactide-co- glycolide) At least one selected may be used.

In addition, the biodegradable polymer in step b) may be contained at a concentration of 10 to 150 mg / ml in the first mixed solution.

In addition, the drug of step c) may be a compound represented by the following formula 1.

[Formula 1]

.

.

In addition, the aqueous medium of step d) may include a surfactant.

The present invention relates to a process for producing biodegradable polymer particles carrying a quinolone antibiotic marbophlogasin by using an extraction evaporation method in a solvent of S / O / W using a minimum process and energy, a high drug loading amount and a filling efficiency, It is possible to minimize the number of administrations of the antibiotics, minimize the antibiotic dose, and maximize the drug efficacy relative to the administration. Thus, the antibiotic-resistant microorganisms Can be minimized, the labor force can be saved, and the productivity can be increased.

FIG. 1 is a process flow chart sequentially showing a method for producing a sustained-release preparation according to the present invention.
FIG. 2 is a photograph of a state in which the drug powder is enclosed in the microparticle formation process by a microscope. FIG. 2 (a) is a 200-fold photograph of Example 2, and FIG.
3 is a SEM photograph of the morphology and surface of the microspheres prepared in Examples 1 to 3 and Comparative Example 2. Fig. a) is Comparative Example 2, b) is Embodiment 1, c) is Embodiment 2, and d) is Embodiment 3.
FIG. 4 is a graph showing the results of measurement of in-vitro drug release behavior of the produced microspheres.

DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments according to the present invention will be described in detail with reference to the accompanying drawings. The terms or words used in the present specification and claims should not be construed to be limited to ordinary or dictionary terms and the inventor shall properly define the concept of the term in order to best explain its invention And therefore should be construed in light of the meanings and concepts consistent with the technical idea of the present invention. In addition, since the embodiments described in the present specification and the configurations shown in the drawings are only the most preferred embodiments of the present invention and do not represent all the technical ideas of the present invention, It is to be understood that equivalents and modifications are possible.

The present invention provides a method for preparing a biodegradable polymer microspheres sustained-release preparation using a solid-in-oil-in-water (solvent intra-extraction-evaporation) method in S / O / W solvent. This method maximizes the amount of encapsulated drug in the microspheres and the efficiency of encapsulation, and makes it possible to prepare a sustained-release preparation which induces sustained and uniform release of the drug.

FIG. 1 is a flowchart schematically showing a method for preparing a sustained-release preparation according to the present invention, and a method for producing the sustained-release preparation of the present invention will be described in detail with reference to FIG.

First, a first mixed liquid is prepared by mixing a non-aqueous solvent and a co-solvent (a).

The non-aqueous solvent is not particularly limited as long as it is capable of dissolving the biodegradable polymer. According to a specific embodiment of the present invention, a non-aqueous solvent having a low boiling point such as a boiling point of 25 ° C to 85 ° C can be used as the non-aqueous solvent. When the boiling point of the non-aqueous solvent is included in the above-mentioned range, it is advantageous in terms of evaporation and drying after the production of the microspheres. In the present invention, examples of the non-aqueous solvent include, but are not limited to, dichloromethane, chloroform, acetonitrile, dimethylsulfoxide, dimethylformamide, ethyl acetate and the like.

The co-solvent is not particularly limited as long as it can dissolve the biodegradable polymer. In the present invention, the co-solvent may be selected from among those which can be mixed with the non-aqueous solvent and the aqueous solvent, have a low solubility of the drug, and have a boiling point lower than that of the non-aqueous solvent. According to a specific embodiment of the present invention, the co-solvent may be at least one selected from the group consisting of methanol, ethanol, acetone, isopropanol, diethyl ether, chloroform, ethyl acetate, acetonitrile and mixtures thereof. .

According to a specific embodiment of the present invention, in the step a), the mixing ratio of the non-aqueous solvent and the co-solvent is 10 to 99% of the non-aqueous solvent and 1 to 90% of the co-solvent based on the volume%. When the amount of the non-aqueous solvent in the first mixed solution is small, the co-solvent is rapidly diffused and extracted with the aqueous solvent and tends to evaporate more rapidly than the non-aqueous solvent, so that uniform formation of the microparticles may become impossible . Further, the microspheres themselves can not be formed, and the drug may not be properly sealed. On the other hand, when the amount of the non-aqueous solvent is increased, the size of the microspheres tends to decrease and the time taken until the microspheres are formed on the aqueous solution by removing the non-aqueous solvent may be prolonged, And the rate of drug encapsulation can be significantly lowered. Accordingly, in the present invention, the mixing ratio of the non-aqueous solvent and the co-solvent in the first mixed liquid is preferably in the range of 20% to 50% of the non-aqueous solvent and 50% to 80% of the co-solvent based on the volume% Do.

Next, a biodegradable polymer and a surfactant are added to the first mixed solution of step (a) to prepare a second mixed solution (b). The biodegradable polymer and the surfactant may be added to the first mixed solution simultaneously or sequentially.

The biodegradable polymer is a conventional polymer used in the production of microspheres that can be used as a drug delivery system, and is biodegradable in vivo and has biocompatibility. In the present invention, the biodegradable polymer may be a biodegradable polymer, a protein such as gelatin, collagen, fibrin, and elastin, a polysaccharide-based natural polymer such as alginate or hyaluronic acid, Or a biodegradable synthetic polymer such as polylactide (PLA), polyglycolide (PGA), poly (lactide-co-glycolide) (PLGA) or poly (lactide-co-glycolide) . According to a specific embodiment of the present invention, the biodegradable polymer is selected from the group consisting of polylactide (PLA), polyglycolide (PGA), poly (lactide-co-glycolide) Glycyrrhizin) glucose, but is not limited thereto.

In one specific embodiment of the present invention, the biodegradable polymer may have a weight average molecular weight of 60,000 or less. For example, poly (lactide-co-glycolide) (50:50) having a molecular weight of about 13,000, poly (lactide-co-glycolide) (50:50) having a molecular weight of about 33,000, poly (Lactide-co-glycolide) (50:50) having a molecular weight of about 20,000, poly (lactide-co-glycolide) (75:25) Can be used. Examples of such a biodegradable polymer include Resomer RG502H, RG503H, RG504H, RG752H and R202H of Boehringer Ingelheim.

In one specific embodiment of the present invention, the degradable polymer may have an intrinsic viscosity of 0.16 to 0.6 dL / g. In the present invention, when the viscosity of the biodegradable polymer is less than 0.1 dL / g, the microparticles filling the drug powder particles are not properly formed or the drug encapsulation rate is significantly reduced. When the viscosity of the biodegradable polymer is more than 0.6 dL / g, The size may be excessively large, and the drug release amount may be small and the desired level of drug efficacy may not be exhibited.

According to a specific embodiment of the present invention, in step (b), the biodegradable polymer is dissolved at a concentration of 10 mg to 150 mg or a concentration of 25 mg to 125 mg, or a concentration of 50 mg to 125 mg per 1 ml of the first mixed solution. When the concentration of the biodegradable polymer is low, the viscosity of the polymer solution is low and the particle size tends to be small when dispersed into the homogenizer. However, since the ratio of the polymer in the microdroplet before the microprojectile formation is small, Therefore, the filling amount of the drug and the filling efficiency are lowered. On the other hand, when the concentration exceeds 150 mg / ml, the size of the microspheres tends to increase as the viscosity of the polymer solution increases.

In one specific embodiment of the present invention, the surfactant is a nonionic surfactant. The content of the nonionic surfactant is 0 to 0.5 parts by weight, or 0.2 to 0.4 parts by weight based on 1 part by weight of the biodegradable polymer.

In the present invention, the nonionic surfactant may be a poloxamer-based polymer. The poloxamer-based polymer contains PEO as a hydrophilic chain, thereby imparting hydrophilicity to the microparticulate matrix and controlling the release rate of the drug. Poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338, Poloxamer 407 and Poloxamer 407 may be used as the poloxamer-based polymer in the present invention. Examples of the poloxamer-based polymer include Poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338, Or a mixture thereof may be used, but the present invention is not limited thereto.

Next, a drug is added to the second mixed solution to prepare an S / O suspension (c).

In the present invention, the drug is not particularly limited and is a physiologically active substance having an ability to act on a living body to cause a biological reaction (change in physiological function, biochemical change, shape change, etc.). According to one specific embodiment of the present invention, the drug is preferably selected from fluoroquinolone antibiotics. Examples of the fluorinated quinolone antibiotics include Nalidixic acid, Ciprofloxacin, Norfloxacin, Ofloxacin, Gatifloxacin, Levofloxacin, Moxifloxacin, Sparfloxacin, Trocafloxacin, Marbofloxacin, and the like, preferably marbofloxacin. Marbophloxacin is a compound represented by the following formula (1) and exhibits an antibiotic effect by inhibiting the action of DNA-gyrase and topoisomerase IV (topoisomerase IV, phase-transforming enzyme).

[Formula 1]

According to a specific embodiment of the present invention, the drug can be added to the first mixed solution together with the biodegradable polymer and the surfactant in step b) to prepare an S / O suspension. In this case, the suspension can be treated with an ultrasonic disintegrator to uniformly disperse the drug in the suspension. This has the advantage that the size of the drug particles is minimized and homogenized and the drug release of the produced microspheres is constantly controlled.

Next, the S / O suspension and the aqueous medium are mixed to prepare an S / O / W emulsion (d).

In the present invention, the aqueous medium includes a water-miscible aqueous solution, and is a solvent that is mixed with a cosolvent without mixing with a non-aqueous solvent. The aqueous medium may be, for example, distilled water, methanol, ethanol, or the like, but is not limited thereto.

According to a specific embodiment of the present invention, the aqueous medium may further comprise a surfactant. The content of the surfactant is in the range of 0.1 g to 10 g, or 3 g to 7 g, or 0.1 (w / v) to 10 (w / v)% or 3 (w / v) w / v). Examples of the surfactant include, but are not limited to, polysorbate, polyethylene glycol, polyvinyl alcohol, poloxamer, span 80, and the like.

In one embodiment of the present invention, the S / O / W emulsion is prepared by mixing the S / O suspension and the aqueous medium at a ratio of 1: 5 to 1: 50, . The mixing ratio is preferably 1:10 to 1:20. When the volume ratio of the S / O suspension to the aqueous medium is less than 1: 5, the space between the fine droplets in the S / O / W emulsion after homogenizing the S / O / W emulsion with a homogenizer is small, Prior to the diffusion and evaporation of larger cosolvents and non-aqueous solvents, fine droplets may aggregate, making it difficult to form uniformly sized microspheres. When the ratio is more than 1:50, the amount of the surfactant to be added is increased, which is disadvantageous in terms of cost.

Next, the co-solvent diffuses and evaporates from the S / O / W emulsion to the aqueous medium to produce microparticles (e).

In step e), the cosolvent is diffused and evaporated into an aqueous medium to induce primary polymer hardening, whereby the drug particles are enclosed in the microspheres. That is, the co-solvent in the micro droplets dispersed in the S / O / W emulsion is rapidly diffused and extracted into the aqueous solvent, and the biodegradable polymer rapidly solidifies to form microspheres. According to a specific embodiment of the present invention, it is preferable that the micro-droplets are vigorously agitated by a magnetic stirrer so that the micro droplets do not aggregate with each other. The formation time of the microspheres may be 10 to 20 minutes. If the microparticle formation time is less than 10 minutes, the biodegradable polymer of the microparticle may be less cured and aggregation may occur between the particles. On the other hand, if it exceeds 20 minutes, the efficiency of enclosing the drug may be lowered.

After formation of microspheres, the aqueous medium and solvent are removed from the S / O / W emulsion and microspheres are obtained.

Next, the microparticles produced in step e) are dried (f).

Secondary polymer curing is induced while evaporating and drying at room temperature and atmospheric pressure to remove some cosolvents and non-aqueous solvents remaining in the microspheres. The evaporation and drying can be carried out for 3 to 6 hours. Through the evaporation and drying process, the structure of the microspheres is strengthened and the drug is completely enclosed in the biodegradable polymer.

According to a specific embodiment of the present invention, the drying may further include a second drying step to completely remove the solvent remaining in the microspheres. The second drying step may be performed at a temperature of 30 ° C to 40 ° C, and the drying time may be 12 to 24 hours. According to a specific embodiment of the present invention, the second drying step may be performed under a vacuum condition. When the drying temperature is less than 30 ° C., the residual solvent may not be completely removed. On the other hand, when the drying temperature exceeds 40 ° C., the glass transition temperature (Tg) of the biodegradable polymer is dried There is a possibility that the microspheres may be transformed during the process, and the drug release control may become impossible due to the structural modification of the microspheres.

As described above, biodegradable polymeric microspheres, which are release agents, can be prepared by using solid-in-oil-in-water (solvent intra-extraction-evaporation) in the S / O / W solvent according to the present invention. The microspheres obtained according to the above method have a high drug loading amount and a high filling efficiency, and thus are excellent in release effect.

In the present invention, the microspheres may contain 40 to 95% by weight, preferably 55 to 75% by weight, of the biodegradable polymer based on 100% by weight of the microspheres. If the biodegradable polymer is contained in an amount of less than 40% by weight, there is a tendency to exhibit multifaceted properties on the surface and inside of the microspheres, resulting in an initial excessive release of the drug or a short release time. On the other hand, if it exceeds 95% by weight, the amount of the microspheres to be administered to the patient or the animal is excessively large, which makes the administration difficult or the administration itself impossible.

In addition, the microspheres may contain 0 to more than 15% by weight, preferably 2 to 10% by weight, of the nonionic surfactant relative to 100% by weight of the microspheres. If the non-ionic surfactant is not included in the microspheres, the matrix of the microspheres is hydrophobic, so that the external solution can not penetrate into the microspheres and the drug in the microspheres can not be dissolved. In this case, as the PLGA is hydrolyzed over time, the drug is released when the release of the drug is sustained or the matrix inside the particle is abruptly destroyed. On the other hand, when the content of the poloxamer-based polymer is more than 15%, the matrix of the microparticles has a strong hydrophilic property, so that the penetration by water rapidly occurs and the drug is rapidly dissolved, Also, the amount of filling of the drug and the filling efficiency may be lowered.

Best Mode for Carrying Out the Invention Hereinafter, preferred embodiments are described to facilitate understanding of the present invention. It should be noted, however, that the following experimental examples are provided only to aid understanding of the present invention, and the present invention is not limited by the following examples.

Example

Examples 1 to 3 Manufacture of Biodegradable Polymeric Microparticles Containing Magok Flocosacin

A biodegradable polymer PLGA (product name: RESOMER® G504H) and a poloxamer-based polymer (product name: Lutrol®127) were mixed with dichloromethane (manufactured by Sigma Aldrich) and acetone (Zhejiang Guobang Pharmaceutical Co., Ltd.) was added thereto and dispersed using an ultrasonic grinder (Fisher Scientific, Sonic Dismembratro Model 500) so as to disperse the S / O suspension. The S / O suspension in which the marbled flocasin was uniformly dispersed was placed in an aqueous solution of 5 (w / v)% polyvinyl alcohol (Sigma Aldrich, 87 to 90% hydrolyzed MW: 30,000 to 70,000) (ULTRA-TURRAX T8) at 5,000-8,000 rpm to obtain an S / O / W emulsion. Thereafter, the emulsion was agitated with a magnetic stirrer (900 rpm) for 10 minutes at room temperature and atmospheric pressure to form microspheres, and then microparticles were collected using Whatman (R) and washed with distilled water for 2-3 times. The microspheres were first dried under normal temperature and normal pressure for 3 hours and then vacuum-dried at 35 DEG C for 12 hours to completely remove the solvent in the microspheres.

Comparative Example

Comparative Example 1

Marbophocosaccharide solutions were prepared by dissolving marbophloxacin powder (drug substance containing microparticles) in PBS buffer (phosphate buffered saline, pH 7.4) at a concentration of 4 mg / ml.

Comparative Example 2

Microspheres were prepared in the same manner as in Example 1, except that no poloxamer was added.

The components and contents used in the above Examples and Comparative Examples are shown in Table 1 below.

ingredient Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 PLGA 500 mg 500 mg 500 mg - 500 mg Pollock Sommer 20 mg 50 mg 100 mg - 0 mg Magoblockocaine 500 mg 500 mg 500 mg 500 mg 500 mg Methylene
Chloride 1 ml 1 ml 1 ml - 1ml Acetone 3 ml 3 ml 3 ml - 3ml Aqueous solution of polyvinyl alcohol 20 ml 20 ml 20 ml - 20ml

Experiments and results

Experimental Example 1: Measurement of the inclusion form of drug powder during the process of forming microspheres

A small amount of the S / O / W emulsion obtained in Example 2 was treated on a slide glass, and the morphology of the drug particles in the fine droplet was observed through a microscope (OLIMPUS, CKX41), and the morphology of the microspheres after the solvent evaporation was confirmed .

FIG. 2 shows the result of the measurement. FIG. 2 is a photograph of the morphology of the drug powder during the process of forming microspheres. FIG. 2 (a) is a photograph 200 times and FIG. 2 (b) is a photograph 400 times magnified.

Experimental Example 2: Measurement of morphology of microspheres

About 5 mg of the microspheres prepared in Examples 1 to 3 and Comparative Example 2 were coated with platinum for 4 minutes using a coater (Quorum Q150 TES, 10 mA), and then subjected to scanning electron microscopy (Tescan Mira 3, LMU FEG-SEM ) And morphology and surface morphology of the microspheres were observed. The measurement results are shown in FIG. 3, wherein a) is Comparative Example 2, b) is Example 1, c) is Example 2, and d) is Example 3. According to the above results, microparticles having sizes of about 80 to 100 mu m were confirmed in each of the examples and the comparative examples, and a smooth surface texture was observed without pores and cracks.

Experimental Example 3: Measurement of the amount of embrocasone encapsulated in the microparticles and the rate of encapsulation

About 30 mg of the microspheres prepared in Examples 1 to 3 and Comparative Example 2 were completely dissolved in 3 ml of chloroform (Sigma Aldrich) and diluted 400-fold to be used as a sample solution. Using a UV-vis meter, The absorbance at the wavelength range was measured to determine the content of magofloxacin contained in the microspheres. The rate of inclusion was calculated as the amount of drug to be injected relative to the dose. The measurement results are shown in Table 2 below.

Drug content (%)
(Drug loading / microparticle weight) x 100 (%) Drug Encapsulation Efficiency (%)
(Drug filling amount / drug input amount) × 100% Example 1 30.57 ± 0.30 62.37 + - 0.61 Example 2 27.48 ± 0.25 57.73 + - 0.52 Example 3 24.26 + - 0.44 53.38 + - 0.97 Comparative Example 2 40.73 + - 0.64 81.46 ± 1.29

Experimental Example 4: Measurement of in-vitro drug release behavior of microspheres

In order to measure the release behavior of the drug, each microparticle prepared in Examples 1 to 3 and Comparative Example 2 was weighed so that the amount of the drug in the particle became 4 mg, and then 1 ml of PBS (phosphate buffered saline, pH 7.4), kept at 37 ° C in an isothermal incubator, and the PBS was replaced at intervals of 48 hours. The PBS extracted for each time period was diluted 40-fold and placed in a disposable cuvette. The absorbance at a wavelength of 335 nm was measured using a UV-vis meter. The concentration of released drug was calculated by converting the absorbance value, and the amount of released drug in each time period was calculated as a cumulative percentage of the total drug (4 mg) of each particle sample. The marbophloxacin solution prepared in Comparative Example 1 was used as a control for this experiment. The measurement results are shown in Fig.

According to the above results, the drug encapsulation efficiency of Comparative Example 2 was somewhat higher than that of Examples 1 to 3, but the drug release amount was less than 40%, and after that, the drug delivery efficiency was drastically decreased and the drug delivery efficiency was lowered. On the contrary, Examples 1 to 3 were confirmed that the release of the drug contained in the microspheres was effectively controlled as compared with Comparative Examples 1 and 2 as microspheres containing a poloxamer surfactant. Depending on the amount of poloxamer added, the rate of drug release can be controlled, and microparticles that continuously release the drug over a period of 2 to 3 days can be prepared.

Claims (11)

a) preparing a first mixed solution by mixing a non-aqueous solvent and a co-solvent;
b) simultaneously or sequentially adding a biodegradable polymer and a Poloxamer polymer to the first mixed solution of step a) to prepare a second mixture;
c) adding a fluoroquinolone antibiotic to the second mixed solution to prepare an S / O suspension;
d) mixing the S / O suspension with an aqueous medium to produce an S / O / W emulsion;
e) producing microparticles from said S / O / W emulsion by diffusion and evaporation of said cosolvent into said aqueous medium; And
f) drying the microspheres produced in step e);
Wherein the fluoroquinolone antibiotic is a fluoroquinolone antibiotic.
The method according to claim 1,
Wherein the non-aqueous solvent has a boiling point of 25 占 폚 to 85 占 폚 and the co-solvent is lower than the boiling point of the non-aqueous solvent.
The method according to claim 1,
Wherein the mixing ratio of the non-aqueous solvent and the co-solvent in the step a) is 20% by volume to 50% by volume of the non-aqueous solvent and 50% by volume to 80% by volume of the co-solvent, ≪ / RTI >
The method according to claim 1,
The poloxamer-based polymer in step b) may be selected from the group consisting of Poloxamer 124, Poloxamer 188, Poloxamer 237, Poloxamer 338, Poloxamer 407, Wherein the antibiotic is at least one selected from the group consisting of a mixture of two or more of the quinolone antibiotics.
The method according to claim 1,
Wherein the poloxamer polymer in step b) is mixed at a ratio of more than 0 to 0.5 part by weight with respect to 1 part by weight of the biodegradable polymer.
The method according to claim 1,
Wherein the biodegradable polymer is selected from the group consisting of polylactide (PLA), polyglycolide (PGA), poly (lactide-co-glycolide) (PLGA), and poly (lactide-co- glycolide) Wherein the method comprises the steps of: (a) preparing a sustained-release formulation containing a fluorinated quinolone antibiotic.
The method according to claim 1,
Wherein the biodegradable polymer in step (b) is contained in a concentration of 10 to 150 mg / ml with respect to the first mixed solution.
The method according to claim 1,
Wherein the drug of step (c) is a compound represented by the following formula (1): < EMI ID =
[Formula 1]

.
The method according to claim 1,
Wherein the aqueous medium in step d) comprises a surfactant, and the surfactant is a mixture of one or more selected from the group consisting of polysorbate, polyethylene glycol, polyvinyl alcohol, poloxamer, and span 80. [ A method for producing a sustained release preparation containing a fluorinated quinolone antibiotic.
The method according to claim 1,
Wherein drying in step f) further comprises steps g) and h) of preparing a sustained release formulation containing a fluorinated quinolone antibiotic.
g) a primary drying step of drying the microparticles produced in step e) under normal temperature and pressure; And
h) a secondary drying step of vacuum drying the microparticles dried in the step g) at a temperature of 30 ° C to 40 ° C.
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