KR102138288B1 - Microcapsule using self-assembling of alpha-synuclein and method for manufacturing the same - Google Patents

Abstract

The disclosed microcapsules include a shell formed by the combination of alpha-synuclein nanocomposites and a loading space formed by the shell. The alpha-synuclein nanocomposite includes alpha-synuclein coated nanoparticles, and the shell includes a beta-fold structure formed by self-binding of alpha-synuclein.

Description

MICROCAPSULE USING SELF-ASSEMBLING OF ALPHA-SYNUCLEIN AND METHOD FOR MANUFACTURING THE SAME

The present invention relates to microcapsules. More specifically, the present invention relates to a microcapsule using self-binding of alpha-synuclein and a method for manufacturing the same.

Alpha-synuclein is an amyloid protein composed of 140 amino acids, and its structure can be roughly divided into three. The first is the amphiphilic N-terminal region (amino acids 1-60) known to interact with the lipid membrane, the second is the hydrophobic middle region (61-95) with a non-Abeta peptide element (NAC) segment. Amino acids), and the third is the highly acidic C-terminal region (amino acids 96-140). This protein belongs to an intrinsically disordered protein (IDP), which has no fixed or ordered three-dimensional structure, and has characteristics of interacting with various substances through its structural plasticity.

This compartmentalization and structural plasticity facilitates electrical bonding with nanoparticles. Alpha-synuclein is also a self-binding protein, and has two self-binding mechanisms, nuclear-dependent assembly and unit assembly. Nuclear dependent assembly models are widely accepted as a mechanism for amyloid production. Once the monomers gather to form a nucleus, the monomers grow around the nucleus and grow into a fibrous structure. The unit assembly model is to rapidly assemble into a beta-screen structure through unit assembly of multimers when exposed to stimuli such as organic solvents or shear forces.

Since alpha-synuclein is a protein existing in vivo, it has high stability as a biomedical use, and can be adsorbed on a hydrophilic surface under acidic conditions, so that it is easy to bind with heterogeneous materials such as antibodies, metals, or functional materials.

Using this property, Korean Patent Registration No. 10-1576204 proposed a PoP nanostructure capable of controlling the release of a drug using alpha-synuclein.

1. Korean Registered Patent 10-1576204

One object of the present invention is to provide a microcapsule that can be used not only as a drug carrier having durability and stability, but also for photothermal treatment.

Another object of the present invention is to provide a method for manufacturing the microcapsules.

However, the problems to be solved by the present invention are not limited to the above-described problems, and may be variously extended without departing from the spirit and scope of the present invention.

The microcapsules according to exemplary embodiments of the present invention for achieving the above object of the present invention include a shell formed by the combination of alpha-synuclein nanocomposites and a loading space formed by the shell. The alpha-synuclein nanocomposite includes alpha-synuclein coated nanoparticles, and the shell includes a beta-fold structure formed by self-binding of alpha-synuclein.

According to one embodiment, the alpha-synuclein nanocomposite includes at least one selected from the group consisting of metal nanoparticles, magnetic nanoparticles, and quantum dots.

According to an embodiment, the microcapsule further includes a transfer material loaded in the loading space.

According to one embodiment, the delivery material includes at least one selected from the group consisting of a hydrophobic material, a hydrophilic material encapsulated by an inverse micelle structure, quantum dots, magnetic nanoparticles, and metal nanoparticles.

According to one embodiment, the microcapsules release the delivery material by proteolytic enzymes or metal ions.

According to one embodiment, the microcapsule is loaded in the loading space, and further includes at least one expanded polymer selected from the group consisting of polycarbonate, polypropylene, and polystyrene.

According to one embodiment, the microcapsule further includes a protective layer surrounding the shell.

According to one embodiment, the protective layer comprises gelatin.

According to one embodiment, the protective layer includes hyaluronic acid.

According to an embodiment, the microcapsules further include a hydrophobic organic solvent loaded in the loading space.

According to one embodiment, the microcapsules, surrounding the shell, further comprises an expansion layer comprising a perfluoropolyether (PFPE).

According to one embodiment, the alpha-synuclein nanocomposite comprises gold nanoparticles.

A method for manufacturing a microcapsule according to exemplary embodiments of the present invention comprises coating an alpha-synuclein on a surface of nanoparticles to form an alpha-synuclein nanocomposite, an aqueous solution of the alpha-synuclein nanocomposite and Mixing a hydrophobic organic solvent to form a picking emulsion, and from the picking emulsion, obtaining a microcapsule comprising a shell formed by the combination of the alpha-synuclein nanocomposites and a loading space formed by the shell. do.

According to an embodiment, the nanoparticles include at least one selected from the group consisting of metal nanoparticles, magnetic nanoparticles, and quantum dots.

According to one embodiment, the hydrophobic organic solvent includes chloroform.

According to one embodiment, the alpha-synuclein nanocomposite is formed in a solution having a pH of 6 to 7.

According to one embodiment, the picking emulsion is formed in a solution having a pH of 6 to 7.

According to one embodiment, in the step of forming the picking emulsion, the delivery material dissolved in the hydrophobic organic solvent is mixed together.

According to one embodiment, the step of obtaining the microcapsules includes the step of evaporating the hydrophobic organic solvent.

According to one embodiment, the method further includes forming a protective layer surrounding the surface of the shell.

According to one embodiment, further comprising the step of binding the antibody to the surface of the shell.

According to the exemplary embodiments of the present invention described above, it is possible to obtain a microcapsule that can be used as a carrier of various materials.

The microcapsules have biocompatibility including alpha-synuclein, have high structural stability by self-binding of alpha-synuclein, can have increased photothermal properties by nanoparticle assembly, and provide various functionalities It is possible to control the release of the loading material.

1 and 2 are diagrams for explaining a method of manufacturing a microcapsule according to an embodiment of the present invention.
3 is a cross-sectional view of a microcapsule according to embodiments of the present invention.
4 is a cross-sectional view showing a mass transfer method using a microcapsule according to an embodiment of the present invention.
5 to 7 are cross-sectional views showing microcapsules according to embodiments of the present invention.
8 is a graph showing the surface plasmon resonance (LSPR) spectrum of gold nanoparticles (20 nm) coated with alpha-synuclein obtained in Experimental Example 1.
9 is a digital photograph before and after sonication of the solution obtained in Experimental Example 2 and a micrograph of a picking emulsion.
10 is a digital photograph before and after sonication of the solution obtained in Experimental Example 3 and micrographs of the picking emulsion.
FIG. 11 is a digital photograph before and after sonication of the solution obtained in Experimental Example 4 and a micrograph of a picking emulsion.
12 is a fluorescence image of the picking emulsion obtained in Experimental Example 5.
13 is a graph measuring the R6G concentration of the picking emulsion exposed to proteases and metal ions in Experimental Example 5.
14 is a graph showing changes in temperature over time in Experimental Example 6.

Hereinafter, a method for extracting and recovering salt according to exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

The present invention can be variously modified and have various forms, and the embodiments described in the text are not intended to limit the present invention to specific disclosed forms, and all modifications included in the spirit and scope of the present invention , It should be understood to include equivalents or substitutes.

The terms used in this application are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, the terms "comprises" or "have" are intended to indicate the presence of features, numbers, steps, actions, elements, parts or combinations thereof described in the specification, one or more other features. It should be understood that the existence or addition possibilities of fields or numbers, steps, actions, components, parts or combinations thereof are not excluded in advance.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person skilled in the art to which the present invention pertains. Terms, such as those defined in a commonly used dictionary, should be interpreted as having meanings consistent with meanings in the context of related technologies, and should not be interpreted as ideal or excessively formal meanings unless explicitly defined in the present application. Does not.

1 and 2 are diagrams for explaining a method of manufacturing a microcapsule according to an embodiment of the present invention.

Referring to FIG. 1, an alpha-synuclein nanocomposite is formed by coating the surface of a nanoparticle with alpha-synuclein. For example, the nanoparticles may be quantum dots, metal nanoparticles, or mixtures thereof. For example, the metal nanoparticles may include gold, silver, platinum, and magnetic metal.

According to one embodiment, the nanoparticles may be gold nanoparticles (AuNP). Gold nanoparticles can easily form an alpha-synuclein bond, and have high biocompatibility. In addition, it is a material suitable for photothermal treatment to be described later.

According to one embodiment, the diameter of the metal nanoparticles may be 100nm or less. When the diameter of the metal nanoparticles is excessively large, stability of the microcapsules may be reduced, or the culture time may be excessively increased.

The alpha-synuclein can adsorb on the surface of the gold nanoparticles to form a coating layer. In other embodiments, the alpha-synuclein may include mutants. For example, alpha-synuclein may have a structure in which some of the amino acids are substituted with cysteine, or cysteine is inserted.

The nanoparticles coated with the alpha-synuclein can be formed in a hydrophilic solution, for example, an aqueous solution, and dispersed therein.

According to one embodiment, in order to obtain a dispersion of nanoparticles coated with stable alpha-synuclein, the pH of the reaction solution is preferably 6 or more, and preferably 6 to 7.

Referring to FIG. 2, the alpha-synuclein-coated gold nanoparticle (αS-AuNP) is formed as a picking emulsion.

Pickering emulsions are emulsions that are stabilized by solid particles where the solid particles are adsorbed at the interface between the two immiscible fluid phases, and the solid particles lower the surface energy of the emulsion and prevent agglomeration.

A hydrophobic organic solvent can be provided to form the picking emulsion. For example, the picking emulsion may be formed by adding the hydrophobic organic solvent to an aqueous solution containing the alpha-synuclein coated gold nanoparticles (αS-AuNP) and mixing using ultrasonic waves.

According to one embodiment, the hydrophobic organic solvent may be used having a higher density than water. Preferably, the hydrophobic organic solvent may include chloroform. Chloroform can increase the dispersion degree of the emulsion and promote the self-bonding of alpha-synuclein. In addition, due to the high density of chloroform, an emulsion is precipitated at the bottom of the aqueous layer, so that the emulsion is not exposed to air and evaporation of chloroform is prevented to prevent fusion, adhesion, or destruction of the emulsion.

Thereby, a microcapsule stabilized from the picking emulsion (having a stable structure by self-binding of alpha-synuclein) can be obtained.

For example, the nanoparticles coated with the alpha-synuclein may be combined into a shape surrounding a sphere to form a shell. Specifically, adjacent nanoparticles are in contact with each other, so that the alpha-synuclein in the shell of the microcapsule can be connected to each other. In the alpha-synuclein, self-binding may be promoted by the chloroform or the like. Accordingly, the shell of the microcapsule may include a beta-screen structure formed through unit assembly of multimers.

In addition, the inside of the microcapsule may be hollow or various desired materials may be loaded as needed.

According to one embodiment, in order to form a spherical microcapsule through the picking emulsion, the pH of the reaction solution is preferably 6 or more, preferably 6 to 7.

According to one embodiment, the microcapsules according to the present invention can be used as a drug delivery system. A drug may be provided when forming the picking emulsion to load the drug in the microcapsule. For example, the drug may be provided dissolved in the hydrophobic organic solvent. For example, the microcapsules formed from the picking emulsion may include a hydrophobic organic solvent and a drug therein.

According to one embodiment, the hydrophobic organic solvent in the microcapsules can be removed. For example, the hydrophobic organic solvent may be volatilized by drying or the like. The hydrophobic organic solvent has high volatility and can be quickly removed, and in the volatilization process, the microcapsules shrink to some extent, thereby narrowing the pores on the surface. Therefore, drugs and the like can be stably loaded in the microcapsules.

The microcapsules according to the present invention are composed of nanoparticles coated with alpha-synuclein, and the nanoparticles are bonded to each other by self-binding of alpha-synuclein. Therefore, even after the organic solvent is removed and the desorption energy disappears, it can maintain its shape. Therefore, it can have high stability as a carrier for drugs and the like.

The microcapsules can be decomposed in a desired region to release a material loaded therein. For example, the microcapsules can be degraded by proteolytic enzymes, proper pH, metal ions, and the like.

According to one embodiment, the microcapsules can be loaded with the drug, it can be decomposed in vivo. For example, when the microcapsule contains an anti-cancer substance and is injected into a cancer patient, the microcapsule may be decomposed by increased matrix metalloproteinase (MMP) in a region where cancer lesions are present. Thus, the loaded anti-cancer material can be provided to the cancer lesion.

In addition, fragments of the degraded microcapsules can be used for photothermal treatment. For example, fragments of the microcapsules may exhibit a photothermal treatment effect by laser irradiation. Since the fragments of the microcapsules contain gold nanoparticles having photothermal properties, treatment of cancer cells or the like can be performed thereby.

Various types of substances may be used as the drug. For example, the drug may include low molecular weight compounds, proteins, recombinant proteins, antibodies, nucleic acids, viruses, and the like.

The microcapsules can be used in a pharmaceutical composition. The pharmaceutical composition comprising the microcapsules may further include a pharmaceutically acceptable carrier, excipient, and diluent in addition to the active ingredients described above for administration.

The carrier, excipients and diluents include lactose, textrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, Polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil.

The pharmaceutical composition may be prepared in various parenteral or oral dosage forms according to known methods. As a representative formulation for parenteral administration, an isotonic aqueous solution or suspension is preferable as a formulation for injection. Injectable formulations can be prepared according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. For example, each component can be formulated for injection by dissolving it in saline or buffer.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, Witepsol, Macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The effective dosage of the pharmaceutical composition may vary depending on the patient's age and gender.

According to one embodiment, the microcapsules of the present invention can change the function or impart various functions by changing the material of the nanoparticles.

3 shows a cross-section of a microcapsule according to embodiments of the present invention.

For example, the shell of the microcapsule is selected from gold nanoparticles coated with alpha-synuclein, quantum dots (QD) coated with alpha-synuclein, and magnetic nanoparticles (MNP) coated with alpha-synuclein. It can contain one.

Quantum dots coated with the alpha-synuclein and magnetic nanoparticles coated with the alpha-synuclein can be obtained in the same manner as the gold nanoparticles coated with the alpha-synuclein.

According to one embodiment, the picking emulsion from a mixture comprising two or more selected from the alpha-synuclein gold nanoparticles, the alpha-synuclein coated quantum dots and the alpha-synuclein coated magnetic nanoparticles By forming, it is possible to obtain a microcapsule made of a combination of the nanoparticles.

The composition of the nanoparticles constituting the shell of the microcapsule may vary depending on the purpose. For example, the quantum dot may be used as a label for optical imaging, bio sensing, and the like. In addition, the magnetic nanoparticles may be used for electromagnetic heating effects or magnetic resonance imaging (MRI).

The microcapsules according to the present invention can load various substances in addition to the hydrophobic drug.

For example, a hydrophilic material can be introduced into the microcapsule by using DOPC or the like to form an inverted micelle structure that captures the hydrophilic material and dissolving it in chloroform or the like to form a picking emulsion.

Further, by forming a picking emulsion by dispersing quantum dots or magnetic particles in chloroform, quantum dots or magnetic particles can be introduced into the microcapsules.

In addition, an antibody (Antibody) can be attached to the surface of the microcapsule if necessary. Since alpha-synuclein is exposed on the surface of the microcapsule, it is possible to induce the binding of the alpha-synuclein to the antibody through a reaction. For example, an antibody for targeting purposes can be attached to the shell of a microcapsule using a photo-active crosslinking agent, Sulfo-SANPAH.

In addition to this, the microcapsules can load/deliver various substances such as pharmaceuticals for animals other than humans, pesticides including pesticides, adhesives, curing agents, and crosslinking agents.

4 is a cross-sectional view showing a mass transfer method using a microcapsule according to an embodiment of the present invention.

Referring to Figure 4, the microcapsules according to an embodiment of the present invention, a shell made of nanoparticles 10 coated with alpha-synuclein, a transfer material 20 loaded in the shell and loaded in the shell It includes an expanded polymer (30).

According to one embodiment, the microcapsules are obtained by forming a picking emulsion by providing a hydrophobic organic solvent and the expanded polymer 30 to a solution containing nanoparticles 10 coated with the alpha-synuclein Can.

According to one embodiment, the expanded polymer 30 may include a hydrophobic polymer having a high coefficient of linear thermal expansion, for example, polycarbonate, polypropylene, polystyrene, and the like.

The microcapsules can be moved to a place to release the delivery material 20 through a suitable method.

After the microcapsule is moved to the target region, light such as a laser may be irradiated to the microcapsule.

When the light is irradiated, the expanded polymer 30 inside the microcapsule is heated by the photothermal effect of the nanoparticles 10. Since the expanded polymer 30 has a high coefficient of linear thermal expansion, it expands by heating, whereby the shell may decompose, or a gap between the nanoparticles 10 may be widened. Accordingly, a substance serving as a solvent or a dispersion medium from the outside may be introduced into the shell, and thus, the transfer material 20 loaded in the shell may be discharged to the outside.

According to the present embodiment, even in a region that cannot provide a substance that decomposes self-binding of alpha-synuclein, such as a protease, an appropriate pH, and metal ions, the delivery material 20 may be emitted using light. .

5 to 7 are cross-sectional views showing microcapsules according to embodiments of the present invention.

Referring to FIG. 5, the microcapsule includes a shell made of nanoparticles 12 coated with alpha-synuclein, a transfer material 22 loaded in the shell, and a protective layer 32 surrounding the shell. According to one embodiment, the microcapsule further comprises a hydrophobic organic solvent in the shell.

According to one embodiment, the protective layer 32 includes a polymer that can be removed or decomposed by heat. For example, the protective layer 32 may include gelatin. Gelatin can dissolve at temperatures above about 35°C and is not soluble in hydrophobic organic solvents. Therefore, it may be suitable for protecting microcapsules loaded with a hydrophobic organic solvent.

The protective layer 32 provides a hydrophobic organic solvent to a solution containing the nanoparticles 12 coated with the alpha-synuclein to form a picking emulsion, and before the hydrophobic organic solvent evaporates, the It can be obtained by coating gelatin or the like.

After the microcapsule is moved to the target region, light such as a laser may be irradiated to the microcapsule. When the light is irradiated, the protective layer 32 is decomposed and removed by the photothermal effect by the nanoparticles 12.

Therefore, the shell made of the nanoparticles 12 coated with the alpha-synuclein is exposed to the outside, and the hydrophobic organic solvent and the delivery material 22 can be released through micropores included in the shell. .

Referring to FIG. 6, the microcapsule includes a shell made of nanoparticles 14 coated with alpha-synuclein, a transfer material 24 loaded in the shell, and a protective layer surrounding the shell.

According to one embodiment, the first region 34a of the protective layer includes a polymer that can be removed or decomposed by heat. For example, the first region 34a of the protective layer may include gelatin.

According to one embodiment, the second region 34b of the protective layer includes a polymer different from the first region 34a. For example, the second region 34b of the protective layer may include a biodegradable material such as hyaluronic acid.

Examples of the present invention are not limited to this, and in addition to the gelatin and hyaluronic acid, may include various materials that can be decomposed/removed in response to an external environment.

Through the combination of the protective layers as described above, it is possible to variously control the conditions of the material release of the microcapsules.

Referring to FIG. 6, the microcapsule includes a shell made of nanoparticles 16 coated with alpha-synuclein and an expansion layer 36 surrounding the shell. The microcapsules may not function as a mass transfer agent. Therefore, the transfer material may not be transferred in the shell. If necessary, a hydrophobic organic solvent may be loaded in the shell.

The expansion layer 36 may include a polymer having a high coefficient of thermal expansion of volume, for example, perfluoropolyether (PFPE).

According to one embodiment, the microcapsules may be dispersed in a hydrogel comprising a structure forming a network structure, for example, fibers. When light is irradiated to the microcapsule, the heat is applied to the expansion layer 36 by the photothermal effect by the nanoparticles 16, and the volume of the expansion layer 36 increases, so that the inside of the network structure is In the gap can vary.

As described above, the microcapsules according to the present invention can be used for various purposes other than mass transfer.

Hereinafter, a microcapsule according to exemplary embodiments and its use and effects will be described in more detail with reference to specific experimental examples.

Experimental Example 1: Preparation of gold nanoparticles coated with protein alpha-synuclein

To obtain gold nanoparticles (αS-AuNP) coated with alpha-synuclein, in a 20 mM Mes(2-(N-morpholino)ethanesulfonic acid) buffer solution (pH 6.5), alpha-mg at a concentration of 1 mg/ml Colloidal gold capped with synuclein and citric acid at a concentration of 5 x 10 ¹³ (5 nm), 5.7 x 10 ¹² (10 nm), 7 x 10 ¹¹ (20 nm), 2 x 10 ¹¹ (30 nm) particles/ml The mixture of nanoparticles (AuNP) in a volume ratio of 1:4 was reacted at 4° C. for 12 hours. Alpha-synuclein not bound to gold nanoparticles was centrifuged at 13,200 rpm for a certain period of time (90 minutes at 5 nm, 60 minutes at 10 nm, 10 minutes at 20 nm, 7 minutes at 30 nm). Removed.

8 is a graph showing the surface plasmon resonance (LSPR) spectrum of gold nanoparticles (20 nm) coated with alpha-synuclein obtained in Experimental Example 1.

Referring to FIG. 8, it can be seen that the nanocomposites were unstable and aggregated in a pH range of 5 or less and 8 or more.

Experimental Example 2: Pickering Emulsion Manufacturing-Gold nanocomposite

After adding chloroform corresponding to 1/10 of the nanocomposite solution obtained in Experimental Example 1 (20 nm gold nanoparticles) to the nanocomposite solution, shaking the sample tube to form a large emulsion, 40 kHz and 100 for 5 seconds The W ultrasonic treatment was performed to form a picking emulsion.

Additionally, instead of chloroform, the same amounts of n-butyl alcohol, ethyl acetate, tert-butyl acetate, toluene and hexane were each added to the nanocomposite solution and the picking emulsion was formed in the same manner.

9 is a digital photograph before and after sonication of the solution obtained in Experimental Example 2 and a micrograph of a picking emulsion. Referring to FIG. 9, when using tert-butyl acetate, chloroform, toluene and hexane, a picking emulsion was formed, but when using chloroform, it can be confirmed that it has a high degree of dispersion. This is because, due to the high density of chloroform, an emulsion is deposited at the bottom of the aqueous layer, so that the emulsion is not exposed to the air and evaporation of chloroform is prevented to prevent fusion, adhesion, or destruction of the emulsion. Thus, when using chloroform, it can be advantageous to further stabilize the picking emulsion and further modify the surface.

Experimental Example 3: Pickering Emulsion Produce - Quantum dots Nanocomposite

Using CdSe quantum dots (QD ₅₂₅ , QD ₆₂₅ ) instead of gold nanoparticles, quantum dot nanocomposites (αS-QD ₅₂₅ , αS-QD ₆₂₅ ) were formed in the same manner as in Experimental Example 1, and the same method as in Experimental Example 2 ( Chloroform) to form a picking emulsion.

10 is a digital photograph before and after sonication of the solution obtained in Experimental Example 3 and micrographs of the picking emulsion. Referring to FIG. 10, it can be seen that in the present invention, a stable picking emulsion can be formed even when quantum dots are used as nanoparticles.

Experimental Example 4: Pickering Emulsion Manufacturing-pH change

In order to confirm the effect of pH on the formation of the picking emulsion, an experiment was performed by changing the pH of the buffer solution (using 20 nm gold nanoparticles and chloroform).

FIG. 11 is a digital photograph before and after sonication of the solution obtained in Experimental Example 4 and a micrograph of a picking emulsion.

Referring to FIG. 11, it can be confirmed that a stable picking emulsion can be formed in a pH range of 6 or more.

Experimental Example 5: Drug loading and release of microcapsules

In order to confirm the loading and release of the load of the microcapsules, rhodamine 6G (R6G) was dissolved in chloroform at a concentration of 10 μg/ml, and a picking emulsion was formed in the same manner as in Experimental Example 2. Next, in order to label the alpha-synuclein located on the surface of the microcapsule, a mouse-derived monoclonal anti-αS antibody was reacted at 37° C. for 2 hours at a dilution ratio of 1:50 and was not bound. After removing the unantibody, the AF488 fluorescent dye-labeled goat-derived anti-mouse antibody was reacted at 37° C. for 1 hour at a dilution ratio of 1:200, and then unbound antibody was removed.

12 is a fluorescence image of the picking emulsion obtained in Experimental Example 5. Referring to FIG. 12, it can be seen that a microcapsule loaded with a drug or the like can be obtained according to the present invention.

Protease (trypsin, cardepsin-B, calpine-1, thrombin, MMP-2 and MMP-9) and metal ions (sodium, potassium, iron, calcium, magnesium and copper) in the picking emulsion containing the microcapsules ) To confirm whether the load was emitted (excited at 552 nm using a spectrophotometer and analyzed for emitted light at 520 nm).

13 is a graph measuring the R6G concentration of the picking emulsion exposed to proteases and metal ions in Experimental Example 5. Referring to FIG. 13, it can be confirmed that the microcapsules of the experimental example released a load from all proteolytic enzymes used, and exhibited a load release effect against copper ions known to specifically bind to alpha-synuclein.

Experimental Example 6: Evaluation of photothermal effect of microcapsules

A laser was irradiated to a gold nanocomposite (αS-AuNP) of the same particle concentration (1.14 x 10 ¹² particles/ml) as a picking emulsion containing the microcapsules of Experimental Example 2 and a comparative example (2000 mW, 532 nm green laser, spaced apart Distance 30cm).

14 is a graph showing the temperature change over time in Experimental Example 6. Referring to FIG. 14, it can be seen that, in the case of the microcapsules formed by the picking emulsion, the initial temperature increase was large compared to the gold nanocomposite particles of the same concentration. This is due to the increase in particle density by the formation of microcapsules, and thus, it can be seen that when the nanoparticles are assembled in the form of microcapsules, an improved photothermal treatment effect can be obtained.

Although described above with reference to the preferred embodiments of the present invention, those skilled in the art may variously modify and change the present invention without departing from the spirit and scope of the present invention as set forth in the claims below. You will understand that you can.

By using the microcapsules according to the exemplary embodiments of the present invention, a general purpose drug delivery system and a drug delivery system having high biocompatibility can be manufactured. In addition, the microcapsules can be utilized as a carrier of various materials. In addition, the microcapsules can be used in the manufacture of stimuli-sensitive smart materials, sensors, soft robotics, and nano-actuators.

Claims (21)

A shell composed of a combination of alpha-synuclein nanocomposites, a loading space formed by the shell, and a delivery material loaded in the loading space, wherein the alpha-synuclein nanocomposite is coated with alpha-synuclein Nanoparticles, the shell comprises a beta-fold structure formed by self-binding of alpha-synuclein,
Microcapsule, characterized in that it further comprises a protective layer surrounding the transfer material and the shell to be loaded in the loading space. The microcapsule according to claim 1, wherein the alpha-synuclein nanocomposite comprises at least one selected from the group consisting of metal nanoparticles, magnetic nanoparticles, and quantum dots. delete The microcapsule of claim 1, wherein the delivery material comprises at least one selected from the group consisting of a hydrophobic material, a hydrophilic material encapsulated by an inverse micelle structure, quantum dots, magnetic nanoparticles, and metal nanoparticles. The microcapsule according to claim 1, wherein the delivery material is released by proteolytic enzymes or metal ions. The microcapsule according to claim 1, further comprising at least one expanded polymer selected from the group consisting of polycarbonate, polypropylene and polystyrene, loaded in the loading space. The microcapsule according to claim 1, further comprising a protective layer surrounding the shell. The microcapsule according to claim 7, wherein the protective layer comprises gelatin. The microcapsule according to claim 7, wherein the protective layer comprises hyaluronic acid. The microcapsule according to claim 7, further comprising a hydrophobic organic solvent loaded in the loading space. The microcapsule of claim 1, further comprising an expansion layer surrounding the shell and including perfluoropolyether (PFPE). The microcapsule of claim 1, wherein the alpha-synuclein nanocomposite comprises gold nanoparticles. Forming an alpha-synuclein nanocomposite by coating the surface of the nanoparticle with alpha-synuclein; And
Mixing the aqueous solution of the alpha-synuclein nanocomposite and a hydrophobic organic solvent to form a picking emulsion; And
And obtaining a microcapsule comprising a shell formed by the combination of the alpha-synuclein nanocomposites and a loading space formed by the shell from the picking emulsion. The method of claim 13, wherein the nanoparticles include at least one selected from the group consisting of metal nanoparticles, magnetic nanoparticles, and quantum dots. The method of claim 13, wherein the hydrophobic organic solvent comprises chloroform. The method of claim 13, wherein the alpha-synuclein nanocomposite is formed in a solution having a pH of 6 to 7. 15. The method of claim 13, wherein the picking emulsion is formed in a solution having a pH of 6 to 7. The method of claim 13, wherein in the step of forming the picking emulsion, the delivery material dissolved in the hydrophobic organic solvent is mixed together. 15. The method of claim 13, wherein the step of obtaining the microcapsules comprises evaporating the hydrophobic organic solvent. The method of claim 13, further comprising forming a protective layer surrounding the surface of the shell. The method of claim 13, further comprising the step of binding the antibody to the surface of the shell.

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