KR20130037955A - Self-fluorescent polymeric nano-complex, contrast agent composition containing the same and method for preparing the same - Google Patents

Abstract

PURPOSE: A self-fluorescent polymeric nanocomposite, a contrast composition containing the same, and a method for manufacturing the same are provided to minimize particle instability and toxicity in vivo. CONSTITUTION: A self-fluorescent polymeric nanocomposite has a self-fluorescent property by crosslinking of a polymeric nanocomposite with a crosslinking agent containing an aldehyde group. The polymeric nanocomposite is prepared by ion bonding of a cationic polymer and an anionic polymer. The anionic polymer is selected from the group consisting of polyglutamic acid, polyacrylic acid, alginate, carrageenan, hyaluronic acid, poly[styrene sulfonate, carboxymethylcellulose, cellulose sulfate, dextran sulfate, heparin], heparin sulfate, poly[methylene-co-guanidine], condroitin sulfate, and a mixture thereof.

Description

Self-fluorescent Polymeric Nano-Complex, Contrast Agent Composition Containing the Same and Method for Preparing the Same}

The present invention relates to a polymer nanocomposite having self-fluorescence properties and consisting of anionic and cationic polymers, a contrast agent composition comprising the same, and a method for preparing the same. It relates to a nanocomposite prepared by reacting with a crosslinker comprising and a method for producing the same.

Molecular imaging is a technique for imaging various phenomena occurring in vivo from the molecular level, and it is a rapidly developing technology field because it provides important clues for disease treatment and metabolic processes in vivo. The biggest advantage of molecular imaging is that the phenomenon that occurs at the cellular or molecular level in the living state of the subject to be studied can be immediately identified through the image and quantitatively analyzed. These types of molecular images include optical imaging using photons in the visible and infrared regions, nuclear imaging using photons generated by radiation reactions, and magnetic resonance generated from the atomic nucleus in the magnetic field. Magnetic resonance imaging (MRI) for imaging signals, and computed tomography (CT) for imaging information obtained by X-ray projection. Among these, optical imaging techniques, which have been used for a long time in the field of molecular / cell biology, are the most versatile in that they have the highest sensitivity of images and can be obtained quickly without exposure to radiation.

Optical images can be classified into bioluminescence imaging and fluorescence imaging. Luminescent images are images of light chemically synthesized by metabolic processes occurring in vivo, and luciferase found in fireflies is used. Fluorescent imaging is a method of labeling and imaging a fluorescent material that absorbs light of a specific wavelength from the outside and emits light of a longer wavelength in a cell, tissue, or living body. Fluorescent molecular images emit the most efficient image signals in the near-infrared region where the absorption of light, scattering, and autofluorescence in living tissues is minimized. Hemoglobin, which is present in large amounts in blood vessels, absorbs light in the visible region, while water and fat in the body absorb light mainly in the infrared region. Therefore, the application of near-infrared fluorescent substance to optical image has the best tissue penetrating power and minimizes actor miscellaneous signal.

With the development of fluorescent imaging equipment, various fluorescent materials are being developed. The types are classified into organic fluorescent dyes and inorganic fluorescent particles. Organofluorescent dyes are already fluorescent materials that are widely used in the overall industrial field and molecular / cell biology research. These organic dyes can be selectively used as necessary because the organic dyes vary depending on the excitation wavelength band and the fluorescence wavelength band emitted. In addition, it is expected to be applied as a targeted fluorescent probe because it is easy to adhere to other materials or biological molecules through chemical bonding. However, most organic dyes are not only difficult to observe for a long time due to low light stability but also have hydrophobic properties, so they may show low solubility in aqueous solution and may be toxic to the living body.

Recently, studies have been conducted to combine such organic dyes with biocompatible biomolecules or biodegradable polymeric materials, but also have side effects due to biocompatibility and binding instability. Inorganic fluorescent particles refer to nano-sized particles having properties similar to semiconductors, commonly called quantum dots, and are recently attracting attention in the field of optical imaging due to their excellent light stability. Since it emits stable fluorescence even after prolonged exposure to light, it is particularly advantageous for observing the differentiation of specific cells or metabolic processes in vivo. These quantum dots emit fluorescence of various colors depending on the size of the particles and have the property of exciting light in a wide wavelength range. Therefore, multi-color imaging that can be observed simultaneously by delivering quantum dots of various sizes to different sites is possible. The disadvantage of quantum dots is that they are synthesized on organic solvents, so they must be surface-modified with a water-soluble substance to be applied in vivo. In addition, since the particle size is larger than that of organic dyes, it is difficult to transfer into cells, and bio stability has not been confirmed since the nuclei of quantum dots are made of heavy metals such as cadmium (Cd) and selenium (Se). Recently, attempts have been made to increase the cell permeability of quantum dots or synthesis using relatively stable metals in vivo instead of heavy metals forming quantum dots. However, currently used fluorescent materials have many problems in terms of biocompatibility or particle stability.

In addition, recently prepared for the production of polymer microbeads (microbeads) having self-fluorescence properties (US Patent Publication No. US20100143259, Wei Wei. Et al . Advanced functional materials; 17: 3153-3158, 2007), but the technology has been reported in this patent, the polymer microbeads are prepared using a special technique called Shirase Porous Glass membrane emulsification technique, and the particle size is limited to 0.2-2 ㎛. There is no choice but to.

Accordingly, an object of the present invention is to provide a nanoparticle composite having autofluorescence characteristics and a method for producing the same, which can effectively dye cells and tissues without using a fluorescent dye.

The present inventors have developed a nano contrast agent that exhibits a self-fluorescence property using no inorganic or organic fluorescent material widely used in the past. In addition, we have developed a method to make nanocomposites that exhibit autofluorescence at the nanoscale level and have multifunctional properties capable of loading cell labeling and other drugs, and to fluoresce effectively in cells and tissues. It confirmed that the characteristic was shown and completed this invention.

The present invention provides a polymer nanocomposite having self-fluorescence, wherein the polymer nanocomposites in which the anionic polymer and the cationic polymer are ionically bonded are crosslinked by a crosslinking agent including an aldehyde group to exhibit self-fluorescence characteristics.

The present invention also provides a contrast agent composition comprising the polymer nanocomposite having the autofluorescence and a pharmaceutically acceptable carrier.

The present invention also comprises the steps of (a) mixing a solution containing an anionic polymer and a solution containing a cationic polymer, to obtain a polymer nanocomposite; And (b) adding a crosslinking agent containing an aldehyde group to the obtained polymer nanocomposite to react, thereby obtaining a polymer nanocomposite having self-fluorescence.

The polymer nanocomposite according to the present invention can be produced easily and reproducibly through ionic assembly using an anionic polymer and a cationic polymer, and the polymer nanocomposite has stability and self-fluorescence properties by a crosslinking agent containing aldehyde. Thus, the polymer nanocomposite having self-fluorescence of the present invention can be used as a new concept of nanocontrast, and can also be used as nanoprobe and drug or gene carrier.

1 is a diagram illustrating a method for synthesizing a polymer ion composite having self-fluorescence characteristics.
Figure 2 is a figure measuring the A) particle size of polygammaglutamic acid (γPGA) / polylysine (PLL) ionic binding complex by DLS; B) It shows the shape and size homogeneity of the particles observed through SEM.
3 is a diagram showing the fluorescence emission spectrum of the poly gamma glutamic acid (γPGA) / poly lysine (PLL) ionic complex after excitation with light of various wavelengths using a fluorescence spectrometer.
4 is a fluorescence image of a polygamma glutamic acid (γPGA) / polylysine (PLL) ionic binding complex and other nano-sized polymer particles using a CCD camera.
FIG. 5 is a diagram observing fluorescence spectra before and after addition of sodium borohydride (NaBH ₄ ) to a polygammaglutamic acid (γPGA) / polylysine (PLL) ionic bond complex. FIG. B) Figure shows the process of reducing the base base by sodium borohydride.
Figure 6 is a polygamma glutamic acid (γPGA) / polyElysine (PLL) ionic complexes treated with HeLa cells (HeLa cells) cancer cells to determine whether the intracellular delivery of particles by fluorescence microscope; B) FACS analysis picture.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein and the experimental methods described below are well known and commonly used in the art.

Self-fluorescence  Having polymer composite

In one aspect, the present invention relates to a polymer nanocomposite having self-fluorescence properties in which anionic polymers and cationic polymers are ion-bonded polymer nanocomposites crosslinked by a crosslinking agent including an aldehyde group (FIG. 1). .

The anionic polymer is not limited as long as it has an anion in a molecule, and is specifically biocompatible and dissociated with an anionic hydroxyl group (-OH), a carboxyl group (-COOH), a sulfate group (-O ₃ SOH, -SH), or It may be a compound having a phosphoric acid group (—O ₃ POH). For example, polyglutamic acid, polyacrylic acid, alginate, carrageenan, hyaluronic acid, polystyrene sulfonate, carboxymethylcellulose Carboxymethylcellulose, Cellulose Sulfate, Dextran Sulfate, Heparin, Heparin Sulfate, Polymethylene-co-guanidine, Condroitin Sulfate ) And complexes thereof may be used alone or in combination of two or more thereof.

In the embodiment of the present invention, poly gamma glutamic acid (γPGA) is used as the anionic polymer. Polygamma glutamic acid is an ultra high molecular weight natural amino acid polymer material biosynthesized by Bacillus subtilis derived from Cheonggukjang, Korea, and has excellent biocompatibility and biodegradability. It is classified as an anionic polymer by a large amount of carboxyl groups present in the constituent amino acid glutamic acid, and is highly reactive with cationic minerals. Therefore, various applications to food additives, health functional foods, cosmetics, and pharmaceuticals are expected. In particular, researches are being actively conducted to apply the drug to the drug delivery system due to its excellent effect of encapsulating hydrophobic drugs, useful proteins and antigens. Induction can form gelled nanoparticles.

The cationic polymer is not limited as long as it has a cation in the molecule, and may specifically be a compound having an amine group (-NH 2) that is biocompatible and has a cation when dissociated. In specific examples, chitosan, polyiminemine, poly (L-lysine), polyallylamine, poly-ornithine, polyvinyl Poly (vinylamine) hydrochloride, polyamido amine (Poly (amido amine), dendrimer (dendrimer), gelatin (Gelatin) and a combination thereof may be used alone or two or more.

In the embodiment of the present invention, polylysine (poly (L-lysine)) is used as the cationic polymer, and the polymer is a biocompatible cationic material composed of lysine which is an essential amino acid. In recent years, research has been actively conducted to apply genes and peptides as transporters for intracellular delivery by using the characteristics that the intracellular delivery is easy due to the cationic nature of polylysine.

The cross-linking agent including the aldehyde group, glutaraldehyde, glyoxal, malonaldehyde, mal aldehyde, succindialdehyde, terephthalaldehyde, formaldehyde, acetaldehyde, propionaldehyde (propionaldehyde), butylaldehyde (butyraldehyde), benzaldehyde (benzaldehyde), cinnamicaldehyde (cinnamaldehyde), 4-methylbenzaldehyde (4-Methylbenzaldehyde), furfural-based material, cationic polymer containing an amine group and One or two or more selected from materials including a reactor capable of inducing seed base (C = N) bonds and complexes thereof may be used.

Crosslinking agents containing aldehyde groups have been widely used in the field of biological tissues in which the structure is fixed and the structure is observed by an optical microscope or an electron microscope. They form chemical bonds between the amine groups of lysine present in the protein to stabilize and fix the tissue structure. As a result of the reaction process, a Schiff's base is formed, in which autofluorescence is generated by an electron transition of a carbon-nitrogen double bond. In the present invention, the fluorescence mechanism is the electron transition of the Schiff's base bond and the carbon-hydrogen double bond.

In one embodiment of the present invention, the anionic polymer is polyglutamic acid, polyacrylic acid, alginate, carrageenan, hyaluronic acid, polystyrene sulfonate, polystyrene sulfonate), Carboxymethylcellulose, Cellulose sulfate, Dextran sulfate, Heparin, Heparin sulfate, Polymethylenecoguanidine (Poly (methylene-co-guanidine sulfonate) )) And Condroitin sulfate, and

The cationic polymer is chitosan, polyethyleneimine, polylysine, polyallylamine, poly-ornithine, poly-ornithine, poly It may be selected from the group consisting of poly (vinylamine) hydrochloride, polyamido amine, dendrimer, and gelatin.

The crosslinking agent is glutaraldehyde, glyoxal, malonaldehyde, malonaldehyde, succindialdehyde, terephthalaldehyde, formaldehyde, acetaldehyde, propionaldehyde, propionaldehyde, and the like. Butylaldehyde, benzaldehyde, benzaldehyde, cinnamaldehyde, 4-methylbenzaldehyde, furfural material And it is apparent to those skilled in the art that it can be selected from a material comprising a cationic polymer comprising an amine group and a reactor capable of inducing a bond base (C = N) bond. Wei Wei. Et al . Advanced functional materials; 17: 3153-3158, 2007)

In the present invention, "nanocomposites" or "nanoparticles" may be particles having a diameter of 1 nm to 1000 nm, preferably 100 nm to 200 nm, and a particle distribution of ± 50 nm. In addition, the nanocomposites of the present invention may be PEGylated with polyethylene glycol. PEGylation improves blood residence time and reduces accumulation in the liver.

The polymer nanocomposite of the present invention uses the ionic self-assembly of the biocompatible polymer electrolyte, which is widely used in optical molecular imaging, and has an advantage of being excellent in solubility in water and biocompatible compared to conventional fluorescent materials. In addition, the cell labeling function can be performed by exhibiting self-fluorescence characteristics through a crosslinking agent including an aldehyde group, thereby replacing a contrast agent composed of existing organic / inorganic fluorescent materials. In addition, it can be loaded with drugs, etc., because it has a human compatibility can be used in medicine, food, cosmetics and the like.

Contrast Composition

In another aspect, the present invention relates to a contrast agent composition comprising the nanocomposite having the above autofluorescence and a pharmaceutically acceptable carrier.

The nanocomposite having self-fluorescence of the present invention is a nanocontrast that exhibits autofluorescence properties without using inorganic or organic fluorescent materials. In addition, it does not exhibit toxicity to cells and exhibits a contrasting effect of anxiety, so it may be commercialized as a safe and excellent contrasting agent.

Carriers used in the contrast agent composition according to the present invention include carriers and vehicles commonly used in the pharmaceutical field, and specifically, ion exchange resins, alumina, aluminum stearate, lecithin, serum proteins (eg, human serum albumin), buffer materials (E.g. partial glyceride mixtures of various phosphates, glycine, sorbic acid, potassium sorbate, saturated vegetable fatty acids), water, salts or electrolytes (e.g. protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride and zinc salts) , Colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose based substrates, polyethylene glycol, sodium carboxymethylcellulose, polyarylates, waxes, polyethylene glycols or wool, and the like. The contrast agent compositions of the present invention may also further comprise lubricants, wetting agents, emulsifiers, suspending agents, preservatives and the like in addition to the above components.

In one embodiment, the contrast agent composition according to the present invention may be prepared as an aqueous solution for parenteral administration, preferably a buffered solution such as Hanks' solution, Ringer's solution or physically buffered saline. Can be used. Aqueous injection suspensions may contain a substrate capable of increasing the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol or dextran.

Another preferred embodiment of the contrast agent composition of the present invention may be in the form of a sterile injectable preparation of a sterile injectable aqueous or oleaginous suspension. Such suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents (e. G., Tween 80) and suspending agents. Sterile injectable preparations may also be sterile injectable solutions or suspensions (eg solutions in 1,3-butanediol) in nontoxic parenterally acceptable diluents or solvents. Vehicles and solvents that may be used include mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, nonvolatile oils are conventionally used as a solvent or suspending medium. For this purpose, any non-volatile oil including synthetic mono or diglycerides and less irritant may be used.

The contrast agent composition according to the present invention may be administered to a living body or a sample, and an image may be obtained by sensing a signal emitted by the polymer nanocomposite having self-fluorescence from the living body or the sample.

The term "sample" as used above refers to tissue or cells isolated from a subject to be diagnosed. In addition, the step of injecting the contrast agent composition into a living body or a sample may be administered via a route commonly used in the pharmaceutical field, parenteral administration is preferred, for example, intravenous, intraperitoneal, intramuscular, subcutaneous or topical route. It can be administered through.

Other uses

The polymer complex having self-fluorescence according to the present invention can be used for nano-probe and drug or delivery vehicle such as separation, diagnosis or treatment of biological molecules.

For example, a pharmaceutically active ingredient may be bound or encapsulated in the polymer nanocomposite of the present invention. The pharmaceutical active ingredient may be, for example, an anticancer agent, an antibiotic, a hormone, an antagonist, an interleukin, an interferon, a growth factor, a tumor necrosis factor, an endotoxin, a lymphokoxy, a urokinase, a streptokinase, a tissue plasminogen activator, a protease inhibitor, an alkyl. Phosphocholine, radioisotope labeled components, cardiovascular drugs, gastrointestinal drugs, nervous system drugs, and the like, but are not limited thereto.

Self-fluorescence  Method for producing a polymer nanocomposite having

In another aspect, the present invention provides a method for producing a polymer composite exhibiting self-fluorescence, comprising the following steps (FIG. 1).

(a) mixing a solution containing an anionic polymer and a solution containing a cationic polymer to obtain a polymer nanocomposite; And

(b) obtaining a polymer nanocomposite having self-fluorescence by adding and reacting a crosslinking agent including an aldehyde group to the obtained polymer composite.

As described above, the existing patent (US 20100143259) was manufactured by using a special technique called “Shirase Porous Glass membrane emulsification technique” in the method of preparing the particles, and the size of the particles was also limited to 0.2-2 μm.

However, according to the present invention, a polymer composite showing self-fluorescence can be prepared more easily and reproducibly using a cationic polymer and an anionic polymer. In addition, the present invention has the advantage that it is easy to obtain a relatively uniform particles of 100-200nm size induce particle formation in the aqueous solution through ionic self-assembly of the ionic polymer.

In the step (a), since the polymer nanocomposite aggregates between molecules by using electrostatic attraction between both polymers, the polymer nanocomposite may be easily prepared by mutually mixing a solution containing an anionic polymer and a cationic polymer, respectively. The reaction conditions are not particularly limited and may be appropriately adjusted depending on the type of polymer. The reaction can be formed at room temperature and it is preferable to mix the polymer solution with stirring.

According to the reaction conditions of step (b) it can form a nanoparticle of a uniform size and shape. In this case, the amount of the anionic polymer and the cationic polymer is preferably 1: 1.8 by weight, and the cationic polymer is contained in an excessive amount. At this time, the particles have a uniform size of about 100-200 nm.

The nanoparticles prepared in step b) may be PEGylated (PEGylation) with polyethylene glycol for application to the living environment. PEGylating the polymer nanocomposites with polyethylene glycol increases the hydrophilicity of the surface of the nanoparticles and prevents recognition from immune functions, including macrophage in the human body that preys and digests pathogens, waste products and foreign influx. Rapid decomposition in the body by the so-called stealth effect can be prevented. Therefore, the retention time in the blood can be improved by the PEGylation. In addition, the polymer nanocomposite PEGylated with polyethylene glycol as described above, unlike the conventional nanoparticles not PEGylated with polyethylene glycol has the advantage that the accumulation in the liver is reduced, the drug delivery composition by enclosing an anticancer agent, etc. It can be used as an advantage. PEGylation can be carried out by methods known in the art. The polyethylene glycol used preferably has a molecular weight between 100 and 5,000, and may have various structures such as linear or branched.

Nanocomposites prepared in the present invention can be separated and purified by means known in the art. An example of the method may be a method of separating the nanocomposites generally produced as precipitates by centrifugation or filtration.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Materials and reagents

Polygamma glutamic acid (γPGA) with a molecular weight of 500 kDa (Bio Leaders, Korea), polylysine (Sigma-aldrich, USA) with a molecular weight of 15-30 kDa, 50% glutaraldehyde (Sigma-aldrich, USA), PEG -NHS (Sunbio, Korea) was prepared.

<1-1> Polygammaglutamic Acid (γ PGA ) / Polylysine ( PLL ) Ionic Binding Complex Formation

Polygamma glutamic acid is dissolved in water at a concentration of 10 mg / ml. Polylysine was dissolved in 10 ml of water at a concentration of 18 mg / ml, and then rapidly added 1 ml of the polyglutamic acid solution prepared in Example 1-2 using a pipette while stirring at 600 rpm, followed by stirring for 2 hours. do. After stirring, 10 μl of glutaraldehyde was added and the mixture was further stirred for 1 hour.

<1-2> Polygamma Glutamic Acid (γ PGA ) / Polylysine ( PLL ) PEGlyation of ionic binding complexes crosslinking

10 μl of glutaraldehyde was added to the polygammaglutamic acid (γPGA) / polylysine (PLL) ionic binding complex solution prepared in Example 1-2, followed by further stirring for 12 hours, followed by a 0.8 μm filter. Filter it out. 250 mg of PEG-NHS was added to 10 ml of the crosslinked polygammaglutamic acid (γPGA) / polylysine (PLL) ionic binding complex solution prepared in Example 1-3 and stirred at 600 rpm for 12 hours. In order to remove the unreacted polymer material and NHS, centrifugation was performed at 5000 rpm for about 15 minutes, the supernatant was discarded, the precipitate was redispersed in tertiary distilled water, and then purified once more by centrifugation. The size of the polygammaglutamic acid (γPGA) / polyElysine (PLL) ionic binding complex was observed by DLS and SEM and the results are shown in FIG.

It was confirmed that the size of the particles measured by DLS in Figure 2 shows a distribution of 100 ~ 200nm (Fig. 2a). As a result of observing the surface shape and size of the particles through the SEM, it was possible to observe the spherical particles of about 100 ~ 200nm similar to the size measured in the DLS (Fig. 2b). When injected into the body, it has a target-oriented characteristic to abnormal tissue in a size suitable for the enhanced permeation and retention (EPR) effect.

Polygamma Glutamic Acid (γ PGA ) / Polylysine ( PLL ) Determination of Activity of Ionic Binding Complexes

<2-1> Polygamma Glutamic Acid (γ PGA ) / Polylysine ( PLL ) Of ionic binding complex Autofluorescence  characteristic

The fluorescence characteristics of the polygammaglutamic acid (γPGA) / polylysine (PLL) ionic complex conjugate solution prepared in Example 1-2 were observed using a fluorescence spectrometer and a CCD camera (FIGS. 3 and 4). . In addition, after raising the pH of the polygamma glutamic acid (γPGA) / polylysine (PLL) ionic binding complex solution prepared in Example 1-2 to 9, NaBH ₄ powder was added by 10% by mass ratio and stirred for 12 hours. Fluorescence was measured (FIG. 5).

In FIG. 3, when the autofluorescent polymer nanoparticle composite was analyzed with a fluorescence spectrometer, the fluorescence emission spectrum having a maximum value at 530 nm was observed when excited with light of various wavelengths.

In FIG. 4, when comparing the polygamma glutamic acid (γPGA) / polylysine (PLL) ionic complex with PLGA or polystyrene nanoparticles, the fluorescence image was examined with a special camera and absorbed light in various wavelengths, thereby fluorescence in a wide wavelength range. The image could be observed.

After the reaction with NaBH ₄ in Figure 5 it can be seen that most of the Schiff's base is reduced to decrease the fluorescence intensity rapidly. NaBH ₄ is known as a substance that suppresses autofluorescence by reducing the base base. (P. Tagliaferro. Et al , Journal of Neuroscience Methods ; 77: 191-197, Wei Wei. meat al . Advanced functional materials ; 17: 3153-3158, 2007)

<2-2> Polygamma Glutamic Acid (γ PGA ) / Polylysine ( PLL ) Nanoparticle Complex Cell Labeling  characteristic

An experiment was carried out to deliver the nanoparticle complex prepared in Example 1-2 into HeLa cells, which are cancer cells. Particles were treated at a concentration of 36 μg / ml, and the degree of cell labeling was observed under a fluorescence microscope. In addition, the particles were treated at various concentrations (2.25, 9, 36 ㎍ / ㎖) and the labeling efficiency was analyzed by FACS analysis.

In FIG. 6, the cells were treated with polygammaglutamic acid (γPGA) / polylysine (PLL) ionic binding complexes to observe fluorescence images. As a result, the cells were effectively labeled with fluorescence at various wavelengths (FIG. 6A). As a result of analyzing the labeled cells by FACS, it was confirmed that the fluorescence intensity tended to increase as the concentration of the treated polygamma glutamic acid (γPGA) / polylysine (PLL) ionic complex was increased.

As described above, specific parts of the present disclosure have been described in detail, and for those skilled in the art, these specific descriptions are merely preferred embodiments, and the scope of the present disclosure is not limited thereto. Will be obvious. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (7)

A polymer nanocomposite having self-fluorescence, wherein the polymer nanocomposites in which anionic polymers and cationic polymers are ion-bonded are crosslinked by a crosslinking agent containing an aldehyde group to exhibit self-fluorescence characteristics. The method of claim 1,
The anionic polymer is polyglutamic acid, polyacrylic acid, alginate, carrageenan, hyaluronic acid, polystyrene sulfonate, carboxymethylcellulose (Carboxymethylcellulose), cellulose sulfate, dextran sulfate, heparin, heparin sulfate, polymethylene-co-guanidine, condroitin sulfate sulfate) and one or two or more kinds selected from the group consisting of these complexes, the polymer nanocomposite having self-fluorescence. The method of claim 1,
The cationic polymer is chitosan, polyethyleneimine, polylysine, polyallylamine, poly-ornithine, poly-ornithine, poly It is characterized in that one or more selected from the group consisting of poly (vinylamine) hydrochloride, poly (amido amine), dendrimer (dendrimer), gelatin (Gelatin) and a complex thereof A polymer nanocomposite having self fluorescence. The method of claim 1,
The cross-linking agent including the aldehyde group, glutaraldehyde, glyoxal, malonaldehyde, mal aldehyde, succindialdehyde, terephthalaldehyde, formaldehyde, acetaldehyde, propionaldehyde (propionaldehyde), butylaldehyde (butyraldehyde), benzaldehyde (benzaldehyde), cinnamaldehyde (cinnamaldehyde), 4-methylbenzaldehyde (4-Methylbenzaldehyde), furfural-based materials, cationic polymer containing an amine group and A material comprising a reactor capable of inducing Schiff base (C = N) bonds, and polymer nanocomposites having one or two or more selected from the group consisting of these. The method of claim 1,
Particle size is 100-200nm, PEGylated (PEGylation), characterized in that the self-fluorescence polymer nanocomposite. A contrast agent composition comprising a polymer nanocomposite having self-fluorescence according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier. A method for producing a polymer composite exhibiting autofluorescence according to claim 1 comprising the following steps:
(a) mixing a solution containing an anionic polymer and a solution containing a cationic polymer to obtain a polymer nanocomposite; And
(b) obtaining a polymer nanocomposite having self-fluorescence by adding a crosslinking agent including an aldehyde group to the obtained polymer nanocomposite.

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