KR20180094518A - A sustained-release injection having microspheres and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a biodegradable polymeric microparticle sustained release preparation carrying a fine powder as a drug and a method for producing such a sustained release preparation. The sustained-release preparation includes a drug-polymer ion composite particle for encapsulating the drug in the biodegradable polymer in the form of a complex with the ionic polymer to delay the release for a long period of time. The manufacturing method according to the present invention is a method of biodegrading a drug-polymer ion complex particle for drug micropowder and emission control using a solid-in-oil-in-water (solvent intra-extraction-evaporation) It is possible to increase the productivity by simple manufacturing process by enclosing the microparticles made of the polymer, maximizing the filling amount and the filling efficiency of the drug in the microparticles, suppressing the initial excessive release, Can be released continuously.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a sustained-release injection microspheres and a manufacturing method thereof,

The present invention relates to a sustained-release preparation containing biodegradable polymer microspheres and a method for producing the same. More particularly, the sustained-release preparation is a drug delivery system in which a drug is loaded in a biodegradable polymer microparticle in a predetermined amount. The drug delivery amount and filling efficiency in the microparticles are maximized, initial excessive release is suppressed, And a method for producing the same.

Alzheimer's disease is the most common symptom of vascular dementia, specific brain disease, and systemic disease, which account for more than 50% of all cases. This disease is named after Alois Alzheimer who discovered a characteristic pathologic finding by observing brain tissue changes in a patient who died of brain cirrhosis in 1906. It accumulates beta amyloid protein between neurons and tangles in neurons, accumulating abnormal proteins and destroying brain cells. In the process of brain cell destruction, the neurotransmitter called Acetylcholine is decreased and symptoms are not revealed yet. Memory impairment, cognitive impairment, and behavioral disturbances. Alzheimer's disease treatment slows the progression of the symptoms, but no underlying cause treatment has yet been developed. Drugs such as Cognex® (Tacrine), Aricept® (Donepezil), Exelon® (Rivastigmine), and NMDA receptor antagonist Namenda® (Memantine), which are currently used to treat or ameliorate these Alzheimer's diseases, include Acetylcholinesterase have. Among them, Rivastigmine is marketed under the trade name of Exelon (R), which is approved by the US FDA in 2007 as a transdermal preparation following oral administration.

Among these drugs, donepezil hydrochloride is used as a cholinesterase inhibitor in the treatment of mild and moderate Alzheimer's disease. It selectively inhibits acetylcholinesterase (AchE), which degrades acetylcholine, so as not to decrease the cerebral neurotransmitter acetylcholine. Donepezil hydrochloride is absorbed in the gastrointestinal tract when taken orally, reaches maximum blood levels 3 to 4 hours after taking, and bioavailability is almost 100%. Donepezil hydrochloride is excreted in the kidney by 72% of the dose, and has a dissipation half-life of about 70 hours, with a plasma concentration of 30 to 75 ng / mL. Many of these side effects have been reported, including nausea and vomiting, which are usually given orally, and patients have memory and thought disorders.

On the other hand, the sustained-release injection formulation refers to an injection formulation which is formulated so that the drug can be continuously and uniformly released while maintaining the 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 multi-emulsion method have disadvantages of high initial release rate, and when the drug as an active ingredient is dissolved in an organic solvent or water, problems such as changes in physical properties of the drug, loss of stability, do.

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. In the active self-healing encapsulation of vaccine antigens in PLGA microspheres, Journal of Controlled Release 165 (2013) 62-74, microparticles were prepared by double emulsion evaporation method and then the active ingredient, vaccine antigen, was loaded through the microspheres Discloses a method of inhibiting the initial release by blocking the micropores by heating the biodegradable polymer to a glass transition temperature (Tg) or more. 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.

SUMMARY OF THE INVENTION Accordingly, it is an object of the present invention to provide a biodegradable polymer microparticle capable of maximally increasing the amount of the drug loaded in the microparticles and the efficiency of encapsulation, and capable of appropriately controlling drug release of the microparticles, and a method for producing the same.

In order to solve the above problems, the present invention provides a sustained-release injectable composition containing biodegradable polymeric microparticles containing a drug, wherein ionic polymer, which is a release-controlling polymer, is ion- complex forming a sustained-release preparation. In the present invention, the biodegradable polymer is slowly hydrated over time after administration of the microspheres, and the drug-polymer complex inside the drug absorbs the electrolyte in the body, and the drug is separated from the complex very slowly.

In general, the release rate of the released microparticle is delayed due to delayed hydration as the crystallinity and molecular weight of the microparticle polymer, the hydrophilicity of the polymer is smaller, and the amount of the drug and the additive is smaller. In particular, for long term sustained release over 1 month, delayed hydration of the microparticulate polymer is preferred and the drug then needs to retard its solubility in body fluids. First, to increase the crystallinity in the microspheres for delaying the hydration of the microspheres, S / O / W type microparticles were prepared to minimize incorporation of drugs and additives into the polymer chains. Secondly, A drug-polymer ion complex which is insoluble in water phase and oil phase is formed and sealed in the microparticulate polymer.

Preferably, the drug-polymer ion complex particles have a size of 0.1 to 10 mu m. The size of such a drug-polymer ion composite particle can be measured, for example, by an electron microscope, and can be expressed in terms of the diameter of the circle by converting the particles seen on the electron microscope into a circle having the same area.

Examples of the ionic polymer that is contained in the microparticle as the drug-polymer ion composite particle for controlling the release include, preferably, a pharmaceutically acceptable polymer such as an anionic polymer such as a condellin sulfate, heparin, polyglutarononic acid , Polyaspartate, polyglutamate, polynucleotide, alginate, hyaluronic acid or a mixture thereof may be used. Examples of the cationic polymer include chitosan, polyethyleneimine, polylysine, polyhistidine, poly Alkyne or mixtures thereof may be used. These release modifying polymers can delay release by absorbing body fluids after injection and separating them from the drug over a prolonged period of time.

The biodegradable polymer according to the present invention is preferably a polylactide (PLA), a polyglycolide (PGA), a poly (lactide-co-glycolide) (PLGA), and a poly (lactide-co- ) ≪ / RTI > glucose may be used.

The drug according to the present invention may be any drug having physical properties that can be contained in the biodegradable microspheres in the manufacturing process according to the present invention. Examples of the drug include haloperidol, flupenthixol, But are not limited to, Fluphenazine, Zuclopenthixol, Pipothiazine, Paliperidone, Olanzapine, Escitalopram, Donepezil, Naltrexone, Risperidone, tacrine, rivastigmine, memantine and the like or a pharmaceutically acceptable salt thereof and a protein or a peptide drug such as insulin or testosterone testosterone, estradiol, medroxyprogesterone, somatropin, lanreotide, octreotide and the like can be used, and anti-cancer The present invention can be applied to therapeutic agents such as leuprolide, tryptorelin and the like.

Preferably, the medicament according to the invention is selected from the group consisting of escitalopram, donepezil, naltrexone, paliperidone or a pharmaceutically acceptable salt thereof, more preferably donepezil, (Donepezil) or a pharmaceutically acceptable salt thereof, and even more preferably Donepezil hydrochloride.

In addition, the microparticle contains 0 to 60% by weight, preferably 20 to 40% by weight, based on 100% by weight of the drug-polymer ion complex. If the microparticle does not contain a polymer that forms an ion complex with a drug, the matrix is formed by dissolving the drug in the microparticle, and the matrix structure becomes non-crystallized, so that the intermolecular force (van der Waals) It rapidly penetrates into the microsphere and the drug inside the microsphere is released within 2-3 weeks, making it difficult to persevere more than one month. However, when an appropriate range of drug-polymer ion complex particles is incorporated, it is present as a non-homogeneous phase between the drug and the microparticle matrix, thereby preventing binding between the biodegradable polymer chains (van der Waals) Slow release occurs. In this case, the amount of incorporation of the drug-polymer complex can be controlled to control the release delay of the microspheres. For example, although depending on the specific type of ionic polymer having a drug and an opposite charge used, about 20% by weight of the matrix is less hydrated and the release occurs over about 2 months, and at 40% by weight, A functional characteristic that can shorten the release rate of the drug can be given within about one month. On the other hand, when the drug-polymer ion complex content exceeds 60%, the drug is excessively exposed to the matrix of the microparticles and rapidly penetrated by the water, resulting in rapid dissolution of the drug complex, The amount of the drug encapsulated and the efficiency of encapsulation may also be lowered.

The present invention also 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.

Specifically, the present invention relates to a method for producing a drug-polymer complex, which comprises the steps of (a) forming a drug-polymer ion complex which is insoluble in a solvent such as an aqueous solvent or a non-aqueous solvent, (B) preparing a first mixed solution S / O suspension in which a biodegradable polymer is dissolved and a drug-polymer ion complex is dispersed, the S / O suspension is added dropwise to an aqueous medium and mixed and stirred (C) producing an S / O / W emulsion, and a curing step (d) in which microparticles are produced by diffusing and evaporating the cosolvent from the S / O / W emulsion into the aqueous medium to provide. In addition, the method according to the present invention may further comprise a step (e) of drying the microparticles produced in the step d).

Illustratively, the preparation method of the present invention can be produced through the following steps.

 (a step) Aqueous solvent Non-aqueous  A step of forming a drug-polymer ion complex which is not dissolved in a solvent to be used

Molecular structural characteristics of the drug are only applicable to ionizable drugs in aqueous solution, and drugs with weak nonionic or ionic properties are not applicable because they can not form ionic complexes.

The drug having primary, secondary, and tertiary amines may have a cation in an aqueous solution, and the pharmaceutically acceptable polymer may be selected from the group consisting of condellin sulfate, heparin, polyglutaronic acid, polyaspartate, Polyglutamate, polynucleotide, alginate, hyaluronic acid, or a mixture thereof. On the other hand, the drug having a carboxyl group, a phosphate group, or a sulfate group may be chitosan, polyethyleneimine, polylysine, polyhistidine, Ionic complexes with polymers such as polyarginine, or mixtures thereof.

The solution is dissolved in an aqueous solution at the same weight ratio, and then dropped under stirring at room temperature while stirring to obtain a white precipitate. Centrifuge at 3000 rpm for 5 minutes, dry the precipitate in a vacuum drier, and pulverize to 0.5 - 10 μm using a ball mill. When the particle size is less than 0.5 μm, it is difficult to grind. When the particle size is more than 10 μm, it is difficult to enclose the particle. A size of 1-3 μm is appropriate to show particle inclusion and release effects.

(Step b) First, a non-aqueous solvent and a co-solvent are mixed to prepare a first mixed solution .

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, non-aqueous solvents 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, the solubility of the drug is low, and the boiling point is lower than that of the non-aqueous solvent. According to a specific embodiment of the present invention, at least one of the co-solvents methanol, ethanol, acetone, isopropanol, diethyl ether, chloroform, ethyl acetate, acetonitrile and mixtures thereof may be used without any particular limitation .

According to a specific embodiment of the present invention, in the step b), 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, preferably, the mixing ratio of the non-aqueous solvent and the co-solvent in the first mixed liquid is in the range of 20% to 50% of the non-aqueous solvent and 50% to 80% .

(Step c) Next, a biodegradable polymer and a drug-polymer ion composite particle are added to the first mixed solution in step a) to prepare a second mixed solution .

The biodegradable polymer and the drug-polymer ion complex 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, a biodegradable synthetic polymer such as polylactide (PLA), polyglycolide (PGA), poly (lactide-co-glycolide) (PLGA), and poly (lactide-co- glycolide) Can be used. 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 added at a concentration of 10 mg to 150 mg, preferably 25 mg to 125 mg, more preferably 50 mg to 125 mg per 1 ml of the first mixture ≪ / RTI > 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.

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

The aqueous medium of the present invention may comprise an emulsifier. 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 emulsifying agent include, but are not limited to, polysorbate, polyethylene glycol, polyvinyl alcohol, poloxamer, span 80, and the like. Preferably, in the present invention, the surfactant is polyvinyl alcohol.

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, there is a greater possibility that the S / O / W emulsion is homogenized with a homogenizer, Prior to diffusion and evaporation of cosolvents and non-aqueous solvents, fine droplets may aggregate, making it difficult to form uniformly sized microspheres. If the ratio is more than 1:50, the amount of the emulsifier to be added increases, which is disadvantageous in terms of cost.

(Step e) Next, the S / O / W emulsion Cooperative  remind In an aqueous medium  As it spreads and evaporates Microsphere  .

In step d), the cosolvent is diffused and evaporated into an aqueous medium to induce a 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 to strongly stir the fine droplets with a magnetic stirrer so that the fine droplets do not aggregate with each other. The formation time of the microspheres may be 1 to 10 minutes. When the microparticle formation time is less than 1 minute, biodegradable macromolecules of the microparticles are less cured and aggregation may occur between the particles. On the other hand, if it exceeds 10 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.

(Step f) Next, the step Microspheres Dry .

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, the biodegradable polymer microspheres can be prepared by using the solvent-in-oil-in-water (S / O / W) solvent intra-extraction evaporation method 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, based on 100% by weight of the biodegradable polymer. 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.

The present invention relates to a process for preparing biodegradable polymer particles carrying a drug-polymer ion complex by using an extractive evaporation method in a solvent of S / O / W using a minimum process and energy, The present invention provides a polymeric microsphere sustained-release preparation capable of exhibiting sustained drug release behavior over a long period of time, and a method of manufacturing the polymeric microsphere sustained release preparation, and more particularly, by controlling the release of ionic complex particles in the microspheres, the drug release of microspheres can be efficiently controlled.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description of the invention, It should not be construed as limited.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view of forming drug-polymer ion composite particles. FIG. Formation of an ionic complex by ionic bond between cationic drug and anionic polymer or anionic polymer and cationic polymer prevents formation of insoluble complex in water phase or oil phase and is not lost during the process so as to enhance loading and enable long term release.
FIG. 2 is a graph showing the results of electron microscopy (SEM) photographs taken before (right) pulverization (left) of donepezil hydrochloride according to Example 1 and pulverized (right) microparticles after donepezil hydrochloride and sodium complexes to be. The particle size is in the range of about 0.5 - 2 um and is insoluble in water and oil solvent, so there is no loss of drug during manufacture and there is an advantage of maximizing loading.
3 is an electron micrograph of the sustained-release preparation prepared in Example 3 according to the present invention before and after drug release. a is a photograph of a microparticle before release, b is a photograph of a particle whose surface is roughly changed by the release of drug-polymer ion complex in an aqueous solution, and c is a photograph in which the shape of the microparticles is collapsed after the drug is completely released.
FIG. 4 is a photograph of the sealed form of the drug-polymer ion composite particle powder during the process of forming microspheres by an optical microscope. A small amount of the S / O / W emulsion obtained in Example 3 of the present invention was treated on a slide glass, It is the shape of the drug particle in the fine droplet observed through the microscope (OLIMPUS, CKX41). In FIG. 2, (a) is a magnification of 200 times, and (b) is a magnification of 400 times.
FIG. 5 is a SEM photograph of the morphology and surface of the microspheres prepared in Examples 3 to 4 and Comparative Examples 3 to 4. FIG. a) is the photographing result of Example 3, b) is the photographing result of Example 4, c) of Comparative Example 3, and d) of Comparative Example 4. Fig.
FIGS. 6 and 7 are graphs showing the results of in vitro drug release behavior measurement of the microspheres prepared in Examples 3 and 4 and Comparative Examples 3, 3-1, 4, and 4-1 according to the present invention.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art to which the present invention belongs.

Example 1: Formation of ion complexes of donepezil hydrochloride and sodium conidiophosphate

100 mg of donepezil hydrochloride was dissolved in 10 ml of distilled water and the pH was adjusted to the pH range of 5-7, which is the neutral region. On the other hand, 100 mg of sodium conidialciferate was dissolved in 10 ml of distilled water and centrifuged at 4000 rpm to prepare a supernatant. The aqueous solution of donepezil hydrochloride was dropped little by little into the dropper while stirring the aqueous solution of the sodium conidialcissylate. When white precipitates were formed as a result of ionic bond formation, stirring was continued for 30 minutes and the reaction was cooled in a 4 ° C refrigerator for 1 hour to sufficiently complete the reaction. The precipitate was centrifuged at 4000 rpm to remove the supernatant, and 20 ml of ethanol was added for dehydration. After mixing and stirring, the supernatant was removed by centrifugation at 4000 rpm, and the precipitate was vacuum-dried at room temperature. The dried ionic complex is pulverized to a size of 10 μm or less using a ball mill.

For the determination of solubility, 100 mg of donepezil complex powder was added to 100 ml of PBS buffer (phosphate buffered saline, pH 7.4), and the mixture was stirred at 600 rpm for 2 hours, 24 hours, 48 hours, After stirring for 96 hours, centrifugation was carried out at 14000 rpm for 10 min, and 1 ml of the supernatant was taken and the concentration was measured at uv 275 nm.

Example 2: Formation of ion complexes of escitalopram oxalate and polyglutaronanic acid sodium

100 mg of escitalopram oxalate was dissolved in 10 ml of distilled water and the pH was adjusted to the pH range of 5-7, which is the neutral region. On the other hand, 100 mg of polyglutarononanoic acid sodium was dissolved in 10 ml of distilled water and centrifuged at 4000 rpm to prepare a supernatant. While stirring the aqueous solution of polyglutarononanoic acid sodium, the aqueous solution of escitalopram oxalate was added little by little to the dropper. When white precipitates were formed as ionic bonds were formed, stirring was continued for 30 minutes and the reaction was cooled in a 4 ° C refrigerator for 1 hour to complete the reaction sufficiently. The precipitate was centrifuged at 4000 rpm to remove the supernatant, and 20 ml of ethanol was added for dehydration. After mixing and stirring, the supernatant was removed by centrifugation at 4000 rpm, and the precipitate was vacuum-dried at room temperature. The dried ion complex is pulverized to a size of 10 μm or less using a ball mill.

For the measurement of solubility, 100 mg of the escitalopram complex powder was added to 100 ml of PBS buffer (phosphate buffered saline, pH 7.4), and the mixture was stirred for 2 hours, 24 hours and 48 hours at 600 rpm on a magnetic stirrer , Stirring for 96 hours, centrifugation at 14000 rpm for 10 min, and then 1 ml of the supernatant was taken and the concentration was measured at uv 275 nm.

Comparative Example  One

100 mg of donepezil hydrochloride powder was added to 100 ml of PBS buffer solution (phosphate buffered saline, pH 7.4), and the mixture was stirred for 2 hours, 24 hours, 48 hours, and 96 hours with a magnetic stirrer at 600 rpm After centrifugation at 14000 rpm for 10 min, 1 ml of supernatant was taken and the concentration was measured at uv 275 nm.

Comparative Example  2

After adding 100 mg of the precipitated palm oxalate powder to 100 ml of phosphate buffered saline (pH 7.4) in a PBS buffer (pH 7.4) for 2 hours, 24 hours, 48 hours, 96 hours After stirring, the mixture was centrifuged at 14000 rpm for 10 min, and 1 ml of the supernatant was taken and the concentration was measured at uv of 275 nm.

Table 1 below shows the solubility of the drug in the PBS buffer solution in Examples 1, 2, and Comparative Examples 1 and 2 over time.

Time (Hr) 2 24 48 96hr Example 1 Concentration (/ / ml) 0 140 564 890 Example 2 Concentration (/ / ml) 0 125 446 970 Comparative Example 1 Concentration (占 퐂 / ml) 980 1023 976 - Comparative Example 2 Concentration (占 퐂 / ml) 965 990 985 -

Table 2 shows the solubility of the drug after 48 hours in aqueous and oily solutions for Examples 1 and 2 and the main ingredient drug.

Distilled water 5% PVA solution Dichloromethane Acetone Example 1 X X X X Example 2 X X X X Comparative Example 1 O O X X Donepezil base X X O O Comparative Example 2 O O X X Eshitalo Palm Base X X O O

(Dissolved O, insoluble X)

It is insoluble in both aqueous phase and oily solvent. When the drug is dissolved in 5% PVA solution and dissolved or dissolved in an organic solvent such as dichloromethane and acetone, the dissolution of the drug in the process of removing the organic solvent causes the disappearance of the drug . Therefore, the drug-polymer ion complex according to the present invention has an advantage of maximizing the loading because it plays a role to prevent such loss.

Fig. 2 is a graph showing the results obtained by preparing ion complexes of donepezil hydrochloride and sodium conidioprolite before pulverization (left) of Donepezil hydrochloride according to Comparative Example 1 and Example 1 in the present invention, and then pulverizing (right) It is an electron microscope photograph. The size of the particles is in the range of about 0.5-2 um.

Example 3: Production of S / O / W type microparticles using biodegradable polymer PLGA, donepezil hydrochloride and sodium complexes cyanine phosphate ion complex (DP-CS complex)

1 g of a biodegradable polymer PLGA (1 g) and a DP-CS complex (1.5 g) were added to 20 ml of a mixture of dichloromethane / acetone (1/3), stirred vigorously to dissolve, S / O suspensions were prepared by dispersing using an ultrasonic mill (Fisher Scientific, Sonic Dismembratro Model 500). The homogeneously dispersed S / O suspension was added to a 200 ml aqueous solution containing 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 over 2 minutes. Thereafter, the emulsion was agitated with a magnetic stirrer (900 rpm) at room temperature and atmospheric pressure for 10 minutes to form microspheres, and then microparticles were collected using Whatman (R) paper and washed with distilled water 2-3 times. The washed microspheres were first dried at room temperature under normal pressure for 3 hours and then vacuum-dried at 35 ° C for 12 hours to completely remove the microspheroidal solvent.

Example 4: Production of S / O / W type microparticles using biodegradable polymer PLGA, escitalopram oxalate and polyglutarononate sodium ion complex (ETP-PGU complex)

1 g of PLGA 1.5 g of EVONIK (product name: RESOMER®RG504H) and 1 g of the ETP-PGU complex were added to 20 ml of a mixture of dichloromethane / acetone (1/3) and dissolved with vigorous stirring. Fisher Scientific, Sonic Dismembratro Model 500) to prepare S / O suspensions.

The following procedure was the same as in Example 3.

Comparative Example 3: Production of S / O / W type microparticles loaded with PLGA and donepezil hydrochloride

1.5 g of PLGA (manufactured by EVONIK, product name: RESOMER® RG504H) and 1 g of donepezil hydrochloride were added to 20 ml of a mixed solution of dichloromethane / acetone (1/3) and dissolved with vigorous stirring. Scientific, Sonic Dismembratro Model 500) to prepare an S / O suspension.

The following procedure was the same as in Example 3.

Comparative Example 3-1: Production of W / O / W type microparticles loaded with biodegradable polymer PLGA and donepezil base

1.5 mg of PLGA was added to 20 ml of a mixed solution of dichloromethane / acetone (1/3) and 1 g of donepezil base, EVOMIK (product name: RESOMER®RG504H), and the mixture was vigorously stirred to dissolve. Scientific, Sonic Dismembratro Model 500) to prepare an S / O suspension.

The following procedure was the same as in Example 3.

Comparative Example 4: Production of S / O / W type microparticles loaded with biodegradable polymer PLGA and escitalopram oxalate

1.5 g of PLGA (manufactured by EVONIK, product name: RESOMER®RG504H) and 1 g of escitalopram oxalate were added to 20 ml of a mixed solution of dichloromethane / acetone (1/3) and dissolved with vigorous stirring. Then, To prepare an S / O suspension.

The following procedure was the same as in Example 3.

Comparative Example 4-1: Production of W / O / W type microparticles loaded with biodegradable polymer PLGA and escitalopram base

1.5 g of PLGA (manufactured by EVONIK, product name: RESOMER®RG504H) and 1 g of escitalopram base were added to 20 ml of a mixed solution of dichloromethane / acetone (1/3), and the mixture was vigorously stirred to dissolve. An ultrasonic grinder To prepare an S / O suspension.

The following procedure was the same as in Example 3.

Experimental Example  One: Microparticle  Measurement of drug powder encapsulation during formation

A small amount of the S / O / W emulsion obtained in Example 3 was treated on a slide glass, and the morphology of the drug particles in the microdroplets was observed through a microscope (OLIMPUS, CKX41). Further, the morphology of the microspheres after the evaporation of the solvent was confirmed.

3 is an electron micrograph of the sustained-release preparation prepared in Example 3 according to the present invention before and after drug release. a is a photograph of the microparticle before release, b is a photograph of particles whose surface has been roughly changed by the drug-polymer ion complex in PBS solution, and c is a photograph in which the shape of the microspheres is collapsed after the drug is completely released.

As shown in FIG. 3, it was confirmed that the DP-CS complex dispersed in the particle state without being dissolved in the microspheres according to the present invention was deformed into the microspheres by swelling in the aqueous solution.

Fig. 4 shows the morphological measurement results in the S / O / W emulsion. FIG. 4 is a photograph of the morphology of the drug powder during the process of forming microspheres. FIG. 4 (a) is a photograph 200 times magnification, and FIG.

As shown in FIG. 4, it can be confirmed that the drug exists in a particulate state in the microspheres in a typical form of S / O / W.

Experimental Example  2: Microsphere  Shape measurement

About 5 mg of the microspheres prepared in Examples 3 to 4 and Comparative Examples 3 and 4 were coated with platinum for 4 minutes using a coater (Quorum Q150 TES, 10 mA), followed by scanning electron microscopy (Tescan Mira 3, LMU FEG-SEM) to observe morphology and surface morphology of the microspheres.

The measurement results are shown in Fig. 5, wherein a) is Example 3, b) is Example 4, c) is Comparative Example 3, and d) is Comparative Example 4. According to the above results, microparticles having a size of about 20 to 60 μm were confirmed in each of the examples and the comparative examples, and smooth surface modification was observed without pores and cracks.

As shown in FIG. 5, although the states of the particles are generally similar, the drug-polymer ion complexes of Example 3 and Example 4 exhibited no smooth surface compared to Comparative Example 3 and Comparative Example 4.

Experimental Example  3: Microsphere  Donepezil hydrochloride Amount of filling  And inclusion rate measurement

30 mg of the microspheres prepared in Examples 3 to 4 and Comparative Example 3, Comparative Example 3-1, Comparative Example 4 and Comparative Example 4-1 were completely dissolved in 3 ml of chloroform (Sigma Aldrich) Diluted 400 times and used as a test solution. The absorbance at a wavelength of 335 nm was measured using a UV-vis meter to determine the content of donepezil and escitalopram 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 3 below.

Drug content (%)
(Drug loading / microparticle weight) x 100 (%) Drug Encapsulation Efficiency (%)
(Drug filling amount / drug input amount) × 100% Example 3 45.7 ± 0.6 82.37 ± 0.2 Example 4 44.4 ± 0.2 78.4 ± 0.4 Comparative Example 3 14.6 ± 0.4 34.5 ± 0.6 Comparative Example 3-1 34 ± 0.2 66 ± 0.5 Comparative Example 4 10.73 ± 0.6 27.3 ± 0.5 Comparative Example 4-1 30 ± 0.6 57 ± 0.7

Experimental Example 4: in vitro  Measurement of drug release behavior

In order to measure the release behavior of the drug, each of the microparticles prepared in Examples 3 to 4 and Comparative Example 3, Comparative Example 3-1, Comparative Example 4 and Comparative Example 4-1 was injected with 40 mg of the drug in the particle The cells were weighed and weighed in 100 ml of PBS (phosphate buffered saline, pH 7.4) and stored at 37 ° C in an isotherm. 1 ml of PBS was diluted 40 times with time, and placed in a disposable cuvette. -vis spectrometer was used to measure the absorbance at a wavelength of 335 nm. The absorbance value was converted to the concentration of the released drug. The cumulative percentage of the released drug in each time period was calculated by comparing the total drug (4 mg) of each particle sample. Donepezil hydrochloride and escitalopram oxalate solutions were used as controls in this experiment. The measurement results are shown in FIGS. 6 and 7. FIG.

According to the results of FIG. 6 and FIG. 7, it can be seen that the initial release is released at an appropriate level in Examples 3 and 4, and the release is continued for about 45 days due to the influence of the ion complex. Comparative Example 3 and Comparative Example 4 were S / O / W type donepezil hydrochloride and escitalopram oxalate, respectively. As a result, the drug was not uniformly distributed between the drug and the matrix polymer, It lasts for about 4 weeks. However, the initial release is too low, the drug is dissolved in the PVA solution during the manufacturing process, resulting in low loading and is not suitable for long-term release over 1 month. In addition, in Comparative Example 3-1 and Comparative Example 4-1 of the commonly used W / O / W type, the binding between the polymer chains was rapidly weakened in a homogeneous phase in which the drug was uniformly dissolved in the PLGA polymer, It has a fast characteristic, and its initial release is fast and its emission duration is very short, about two weeks.

Claims (6)

A sustained-release injection containing biodegradable polymer microparticles comprising a drug, wherein the drug is dispersed in the form of a drug-polymer ion complex particle. The sustained-release formulation according to claim 1, wherein the drug-polymer ion complex particle has a size of 0.1 to 30 μm.  The drug-polymer complex according to claim 1, wherein the drug-polymer ion complex is formed of an anionic drug and a cationic polymer which is cationic in an aqueous solution and is chitosan, polyethyleneimine, polylysine, polyhistidine, polyarginine or a mixture thereof Features a sustained-release injection. The pharmaceutical composition according to claim 1, wherein the drug-polymer ion complex is selected from the group consisting of a cationic drug and a cationic drug that exhibits an anion in an aqueous solution, such as a carrier protein, heparin, polyglutaronic acid, polyaspartate, polyglutamate, polynucleotide, alginate ), Hyaluronic acid, or a mixture thereof. The biodegradable polymer of 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) ≪ / RTI > or a combination thereof. The pharmaceutical composition according to claim 1, wherein the drug is selected from the group consisting of Haloperidol, Flupenthixol, Fluphenazine, Zuclopenthixol, Pipothiazine, Paliperidone, (Olanzapine), Escitalopram, Donepezil, Naltrexone, risperidone, Tacrine, Rivastigmine, Memantine insulin, ), Testosterone, estradiol, medroxyprogesterone, somatropin, lanreotide, octreotide, leuprolide, tryptophan, tryptophan, A sustained-release injectable formulation which is triptorelin or a pharmaceutically acceptable salt thereof.

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