KR101624842B1 - Complex of Hyaluronic Acid-Metal Oxide Nano Particle, the Method for Manufacturing of the Complex and uses thereof - Google Patents

Abstract

The present invention relates to a manufacturing method of spherical hyaluronic acid-nanoparticle complex, spherical hyaluronic acid-nanoparticle complex, a MRI contrast medium, and a drug carrier. More particularly, the manufacturing method of spherical hyaluronic acid-nanoparticle complex comprises the steps of: (a) manufacturing a mixture solution by adding metal oxide nanoparticle aqueous solution, hyaluronic acid, a cross-linker, and a surfactant to oil; and (b) adding acid and a coupling agent to the mixture solution, and mixing the resulting mixture. The spherical hyaluronic acid-nanoparticle complex manufactured according to the present invention can be used as a MRI contrast medium for diagnosis and as a drug carrier.

Description

TECHNICAL FIELD The present invention relates to a hyaluronic acid-metal oxide nanoparticle composite, a method for producing the same,

The present invention relates to a composite comprising hyaluronic acid and metal oxide nanoparticles bound thereto, a method for preparing the same, and a use thereof. More particularly, the present invention relates to a hyaluronic acid-metal oxide nanoparticle having metal oxide nanoparticles inserted into hyaluronic acid, Particle composite, a method for producing the same, a contrast agent, and a use as a drug delivery system.

Hyaluronic acid (HA) is a polysaccharide that exists in the extracellular matrix, connective tissue, joints, and umbilical cord of a higher animal. Glucronic acid and N-acetylglucosamine And has a molecular weight of about 100,000 to 10,000,000. It is known that hyaluronic acid plays a role as a lubricant in the joints, preventing bacterial invasion, penetration of toxins, controlling cell adhesion and inflammatory response. Since hyaluronic acid is a substance in vivo, it does not cause immunity, has excellent biocompatibility and can be biodegraded, and is suitable as a carrier of tissue engineering or drug delivery system.

Meanwhile, the size of the object to be treated in nanotechnology is 1 to 100 nm, which controls the size of nanoparticles to control the physical and chemical properties of the nanoparticles. In the bio field, nanoparticles can be used for measurement and drug delivery. In addition, in the case of nanoparticles, the amount of light energy absorbed and emitted differs depending on the size of the particles, and nanotechnology such as nanometer measurement technology has been developed using the same. Nanometer measurement technology has a wide range of technology and is actively used in measuring fields such as scanning tunneling microscope and atomic force microscope.

The inventors of the present invention have conducted research on preparing spherical composites containing metal oxide nanoparticles in hyaluronic acid which is a biocompatible polymer. As a result, it has been found that when the amount of the surfactant added is controlled, the structure of the hyaluronic acid- For the first time.

Through this, it is possible to successfully carry out the research to develop the contrast agent that can reduce the side effect of MRI and improve the accuracy of the image, and to develop it as a drug delivery vehicle capable of simultaneously conducting diagnosis and targeting treatment.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method for producing a hyaluronic acid-nanoparticle spherical composite containing metal oxide nanoparticles.

Another object of the present invention is to provide a hyaluronic acid-nanoparticle spherical composite containing metal oxide nanoparticles.

Another object of the present invention is to provide an MRI contrast agent containing hyaluronic acid-nanoparticle spherical complex containing metal oxide nanoparticles.

Another object of the present invention is to provide a drug delivery system in which a spherical composite of hyaluronic acid-nanoparticles containing metal oxide nanoparticles is combined with a drug.

The present invention has been made in view of the above problems, and it is an object of the present invention to provide a method of manufacturing the same.

According to an aspect of the present invention, there is provided a method for preparing a hyaluronic acid-nanoparticle spherical composite,

(a) preparing a mixed solution by adding an aqueous metal oxide nanoparticle solution, hyaluronic acid, a crosslinking agent and a surfactant to an oil

(b) adding an acid to the mixed solution, adding a coupling agent, and stirring

.

According to an aspect of the present invention, there is provided a hyaluronic acid-nanoparticle spherical composite according to one embodiment of the present invention, wherein a hyaluronic acid polymer cross-linked by a crosslinking agent comprises a plurality of metal oxide nanoparticles in a spherical hyaluronic acid spherical body A spherical composite of hyaluronic acid-nanoparticles.

According to an aspect of the present invention, there is provided an MRI contrast agent comprising, as an active ingredient, a hyaluronic acid-nanoparticle spherical composite comprising a plurality of metal oxide nanoparticles in a hyaluronic acid spherical body .

According to an aspect of the present invention, there is provided a drug delivery system comprising a hyaluronic acid-nanoparticle spherical composite having a plurality of metal oxide nanoparticles in a hyaluronic acid spherical body, Lt; / RTI >

The details of other embodiments are included in the detailed description and drawings.

The effects of the present invention are as follows.

(i) preparing a mixed solution by adding (a) an aqueous solution of a metal oxide nanoparticle, hyaluronic acid, a crosslinking agent and a surfactant to an oil; And (b) adding an acid to the mixed solution, and then adding a coupling agent and stirring the mixed solution. The hyaluronic acid-nanoparticle spherical composite and the hyaluronic acid-nanoparticle spherical composite are provided.

(ii) In the present invention, by inserting metal oxide nanoparticles into hyaluronic acid, which is a biocompatible polymer, the hyaluronic acid-nanoparticle spherical complex can be used not only as an MRI contrast agent for diagnosis, but also as a drug Lt; / RTI >

The effects according to the present invention are not limited by the contents exemplified above, and more various effects are included in the specification.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing the basic unit structure of a hyaluronic acid polymer constituting a hyaluronic acid-iron oxide nanoparticle composite according to an embodiment of the present invention. FIG.
FIG. 2 is a flow chart of the manufacturing process of iron oxide nanoparticles according to one embodiment of the present invention.
FIG. 3 is a flowchart illustrating a method of manufacturing a spherical hyaluronic acid-iron oxide nanoparticle composite according to an embodiment of the present invention.
4 is a graph showing the measurement of the size of iron oxide nanoparticles according to the change of NH ₄ OH according to an embodiment of the present invention.
5 and 6 are SEM images of a spherical hyaluronic acid-iron oxide nanoparticle composite according to an embodiment of the present invention.
7 is data on the content of constituents of elements contained in a spherical hyaluronic acid-iron oxide nanoparticle composite according to an embodiment of the present invention.
8 is a fluorescence microscope image of a spherical hyaluronic acid-iron oxide nanoparticle composite according to an embodiment of the present invention, showing an image of a composite structure when 250 ㎕ of Span-80 was added.
9 is a fluorescence microscope image of a spherical hyaluronic acid-iron oxide nanoparticle composite according to an embodiment of the present invention, when 500 Sp of Span-80 is added.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention and the manner of achieving them will become apparent with reference to the embodiments described in detail below with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

Although the first, second, etc. are used to describe various components, it goes without saying that these components are not limited by these terms. These terms are used only to distinguish one component from another. Therefore, it goes without saying that the first component mentioned below may be the second component within the technical scope of the present invention.

In one aspect of the present invention, the present invention provides a method for producing a hyaluronic acid-nanoparticle spherical composite,

(a) preparing a mixed solution by adding an aqueous metal oxide nanoparticle solution, hyaluronic acid, a crosslinking agent and a surfactant to an oil

(b) adding an acid to the mixed solution, and then adding a coupling agent and stirring the mixed solution.

In another aspect of the present invention, there is provided a hyaluronic acid-nanoparticle spherical composite in which a plurality of metal oxide nanoparticles are contained in a spherical hyaluronic acid spherical hyaluronic acid polymer cross-linked by a crosslinking agent.

FIG. 1 shows the structure of a basic unit of a hyaluronic acid polymer used in a method for producing a porous membrane according to an embodiment of the present invention. Hyaluronic acid is a biopolymer repetitively containing disaccharide units of glucuronic acid and N-acetylglucosamine, and the molecular weight ranges from 1000 to 10,000,000 Da. Depending on the molecular weight of hyaluronic acid, hyaluronic acid below 1,000,000 Da can be used mainly as a raw material for foods, cosmetics, eyedrops and the like. The higher polymer hyaluronic acid can be used as a raw material for injecting knee joints, and the ultrahigh molecular weight hyaluronic acid having a molecular weight of 6,000,000 Da or more is used as an ophthalmic surgical assistant, an anti-adhesion agent, etc., and it is very important to select hyaluronic acid having an appropriate molecular weight. The molecular weight of the hyaluronic acid used to prepare the porous nanomembrane of the present invention is about 100,000 to 1,000,000 Da, more preferably, 600,000 Da to 900,000 Da, and hyaluronic acid having a molecular weight is used.

In one embodiment of the present invention, the metal oxide nanoparticle aqueous solution may include iron oxide nanoparticles. In the iron oxide nanoparticles of iron oxide it is its molecular formula can include _{FeO, Fe 3 O 4, Fe} 4 O 5, Fe 2 O 3, Fe 2 O 3, Fe 2 O 3, Fe 2 O 3, Fe 2 O 3 , etc. have. Further, the metal oxide nanoparticles may be zinc oxide (ZnO) nanoparticles. The metal oxide nanoparticles may also be manganese oxide nanoparticles. The molecular formula of the manganese oxide can include _{MnO, Mn 3 O 4, Mn} 2 O 3, MnO 2, Mn 2 O 7. The metal oxide nanoparticles may also be chromium oxide nanoparticles. The molecular formula of chromium oxide may include CrO, Cr ₂ O ₃ , CrO ₂ , CrO ₃ , CrO ₅ , and the like. The metal oxide nanoparticles may also be gadolinium oxide (Gd ₂ O ₃ ) nanoparticles. The metal oxide nanoparticles may also be cobalt oxide nanoparticles. The molecular formula of cobalt oxide may include CoO, Co ₂ O ₃ , Co ₃ O ₄ , and the like. The metal oxide nanoparticles may also be nickel oxide nanoparticles. The molecular formula of nickel oxide may include NiO, Ni ₂ O ₃ , and the like.

In one embodiment of the present invention, the metal oxide nanoparticles may have an average diameter of 30 to 50 nm. The metal oxide nanoparticles can be prepared in aqueous solution conditions and various metal precursors can be used to produce the metal oxide nanoparticles.

As the metal precursor, various metal salts can be used, for example, metal chloride, metal acetylacetonate, metal bromide, carbonate, fluoride ), Metal iodides, metal nitrates, metal sulfates, metal oleates, metal formates, hydrates thereof, or mixtures thereof, and the like. But it is not so limited. More preferably, a metal chloride can be used.

In one embodiment of the present invention, in order to prepare the iron oxide nanoparticle aqueous solution (Fe ₃ O ₄ )

(i) the step of using a ₂ · H ₂ O and FeCl ₃ · H ₂ O was added to FeCl water heating and stirring;

(ii) adding NH ₄ OH to the stirred aqueous solution, followed by heating and stirring; And

(iii) cooling the aqueous solution prepared through step (ii), it is possible to prepare an aqueous iron oxide nanoparticle solution.

The size of the metal oxide nanoparticles prepared according to an embodiment of the present invention may be 30 to 50 nm in average diameter. The metal oxide nanoparticles of the size are contained or fixed in the hyaluronic acid spheres to form a complex. The iron oxide, manganese oxide, chromium oxide, zinc oxide, nickel oxide, cobalt oxide or gadolinium oxide nanoparticles contained in the hyaluronic acid complex may be fixed or contained in a plurality of nanoparticles. More preferably, the nanoparticles contained in the hyaluronic acid complex are iron oxides.

The size of the hyaluronic acid-nanoparticle spherical composite prepared according to one embodiment of the present invention may be 50-1000 mu m in average diameter. More preferably, the spherical composite has an average diameter of 100 to 300 mu m.

A cross-linking agent is added to crosslink the hyaluronic acid chain to prepare a complex of hyaluronic acid-metal oxide nanoparticles of spheres. The purpose of the cross-linking process is for the process of binding the polymer strands to the other polymer strands by repeating sequences of glucuronic acid and N-acetylglucosamine monomers constituting hyaluronic acid. An amine group may be introduced to bond to a carboxylic acid functional group of glucuronic acid to form an amide bond and adipic acid dihydrazide (ADH) may be added as a crosslinking agent, Hexamethylene diamine, cystamine, succinic dihydrazide, hexamethylene diamine, isophthalic-dihydrazide, or malonic acid dihydrazide. Malonic dihydrazide and the like can be used. Preferably, the cross-linking agent is adipic acid dihydrazide (ADH).

In the hyaluronic acid-nanoparticle spherical composite according to one embodiment of the present invention, a surfactant is added for the spherical shape to be successful. The nonionic surfactant may be a nonionic surfactant such as polysorbate, polyoxyethylene glycol alkyl ethers, or the like. The nonionic surfactant may be a nonionic surfactant, , Octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether, polyoxypropylene glycol alkyl ether, glucoside alkyl ether, Decyl glucoside, lauryl glucoside, octyl glucoside, polyoxyethylene glycol octylphenol ether, Triton X-100, polyoxyethylene glycol octylphenol ether, Polyoxyethylene glycol alkylphenol ether, Nonoxynol-9, Glycerol alkyl esters, and the like (Glycerol alkyl ester), glyceryl laurate (Glyceryl laurate).

More preferably, the polysorbate according to one embodiment of the present invention is selected from the group consisting of sorbitan monolaurate (Span-20), sorbitan monopalmitate (Span-40), sorbitan monostearate (60) Span-60), Sorbitan tristearate (Span-65) and polysorbate 80 (sorbitan monooleate, Span-80). Even more preferably, the polysorbate according to one embodiment of the present invention is polysorbate 80 (sorbitan monooleate, Span-80).

In addition, in the hyaluronic acid-nanoparticle spherical composite of the present invention, the addition amount of the surfactant should be adjusted so that the spherical shape is successful.

According to one embodiment of the present invention, the addition ratio of the surfactant to be added to achieve spherical shape in the hyaluronic acid-nanoparticle spherical composite is 7 to 11 equivalents relative to the equivalent of the hyaluronic acid, the crosslinking agent and the coupling agent to be added, Preferably 8 to 10 equivalents.

A coupling agent is required for the amide bond between carboxylic acid and amine. Unless a coupling agent is added, a salt is formed by an acid-base reaction between a carboxylic acid and an amine to form an amide bond, and if heated to a high temperature, the dehydration reaction proceeds to form an amide bond , It may affect the physical properties of hyaluronic acid, and it is more preferable to add a coupling agent so as to easily form an amide bond for reaction under milder conditions. The coupling agent is preferably a carbodiimide reagent such as 1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), and diisopropylcarbodiimide (DIC) and dicyclohexylcarbodiimide (DCC) may be used instead of the coupling agent.

According to another aspect of the present invention, there is provided a nuclear magnetic resonance imaging (MRI) apparatus including an hyaluronic acid-nanoparticle spherical composite comprising a plurality of metal oxide nanoparticles in a hyaluronic acid spherical body as an active ingredient, ) Contrast medium. From this, the imaging function of the MRI is included. When nanoparticle particles are included as an MRI contrast agent, it may be more preferably used as an MRI contrast agent for iron oxide. The manufacturing process is fairly simple, and the size of nanoparticle particles can be controlled, making it easy to diagnose.

The main chain hyaluronic acid used according to one embodiment of the present invention preferably has a molecular weight of 600,000 to 900,000 Da. When the molecular weight is too small, migration to the blood vessel tissue may occur when the contrast agent is administered. If the molecular weight is too large, the problem may occur that the body may be kept in the body for a long time for a long time.

The average diameter of the complex in the MRI contrast agent containing the hyaluronic acid-nanoparticle spherical complex used as an active ingredient according to one embodiment of the present invention is 50 to 1000 mu m, more preferably 100 to 300 mu m.

In another aspect of the present invention, there is provided a drug delivery system in which a drug of an effective ingredient is bound to a hyaluronic acid-nanoparticle spherical composite. The material to be bound to the hyaluronic acid-nanoparticle spherical composite of the present invention is not limited and includes various materials showing therapeutic efficacy. According to a preferred embodiment of the present invention, the substance to be delivered is a protein, a nucleic acid molecule, a saccharide, a lipid, or a nanoparticle. The protein delivered by the substance delivery system of the present invention may be insulin, insulin-like growth factor 1 (IGF-1), growth hormone, erythropoietin, granulocyte-colony stimulating factors (G-CSFs), granulocyte interferon beta, interferon gamma, interleukin-1 alpha and beta, interleukin-3, interleukin-4, interleukin-6, interleukin-2, EGFs (epidermal growth factors), calcitonin ), Vascular Endothelial Cell Growth Factor (VEGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), adrenocorticotropic hormone (ACTH), transforming growth factor beta (TGF-beta), bone morphogenetic protein But are not limited to, TNF (tumor necrosis factor), atobisban, buserelin, cetrorelix, deslorelin, desmopressin, dynorphin A (1-13), elcatonin, eleidosin, eptifibatai eptifibatide, growth hormone releasing hormone-II (GHRH-II), gonadorelin, goserelin, histrelin, leuprorelin, lypressin, But are not limited to, octreotide, oxytocin, pitressin, secretin, sincalide, terlipressin, thymopentin, thymosine, The compounds of the present invention may be used in combination with other therapeutic agents such as triptorelin, bivalirudin, carbetocin, cyclosporin, exedine, lanreotide, luteinizing hormone-releasing hormone (LHRH), naparelin ), Parathyroid hormone, pramlintide, T-20 (enfuvirtide), thymalfasin and chiconotide. The nucleic acid molecules that can be carried by the hyaluronic acid-nanoparticle spherical complex of the present invention include, for example, DNA, DNA aptamers, RNA aptamers, ribozymes, miRNA, antisense oligonucleotides, siRNA, shRNA, But is not limited thereto.

The material transported by the drug delivery vehicle having the drug of the active ingredient bound to the hyaluronic acid-nanoparticle spherical composite of the present invention is preferably a pharmaceutical composition, and examples thereof include an anti-inflammatory agent, an analgesic agent, an antiarthritic agent, An antihypertensive agent, an antimicrobial agent, an antibiotic agent, an appetite suppressant, an analgesic agent, an anticholinergic agent, an antihistamine agent, an anti-depressant, an antipsychotic agent, an antidepressant, an antidepressant, , Anti-migraine agents, hormones, coronary blood vessels, cerebrovascular or peripheral vasodilators, contraceptives, antithrombotic agents, diuretics, antihypertensive agents, therapeutic agents for cardiovascular diseases, cosmetic ingredients (for example, wrinkle removers, skin aging inhibitors and skin lightening agents) But is not limited thereto. In addition, according to one embodiment of the present invention, the pharmaceutically acceptable carrier to be contained in the pharmaceutical composition is one commonly used in the present invention and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, But are not limited to, calcium, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil , But are not limited thereto. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

In the drug delivery system in which the drug is bound to the hyaluronic acid-nanoparticle spherical composite used according to an embodiment of the present invention, the average diameter of the complex is 50 to 1000 mu m, more preferably 100 to 300 mu m.

Example

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 describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example 1: Reagent preparation

Hyaluronic acid-nanoparticles The reagents and solvents required to prepare spherical complexes were prepared. In the following Tables 1 and 2, reagents and solvents for preparing the hyaluronic acid-iron oxide nanoparticle complexes are summarized, and the entire experimental procedure is shown in FIG.

Reagent type

Molecular structure
Molecular Weight
amount
equivalent weight Hyaluronic acid (HA)

730 kDa
50 mg
One Adipic acid dihydrazide (ADH)

174.2 g / mol
25 mg
One EDC

191.7 g / mol
25 mg
One Span-80

1310 g / mol
500 μl
9

Solvent type

Molecular Weight
amount
Mole ratio
FeCl ₃ .6H ₂ O

270.30 g / mol

0.606 g (2.2 mmol)
2
FeCl ₂ .4H ₂ O

198.81 g / mol
0.226 g (1.1 mmol)
One
NH ₄ OH

35.05 g / mol
0.7 ml (5.3 mmol)
5

Example 2: Preparation of aqueous solution of iron oxide nanoparticles

1) After adding 30 ml of DW to a 250 ml three-necked flask, 0.226 g (2.2 mmol) of FeCl ₃ .6H ₂ O and 0.226 g (1.1 mmol) of FeCl ₂ .4H ₂ O were added to the flask. (750 rpm) < RTI ID = 0.0 >

2) Add 0.7 ml of ammonium hydroxide (NH ₄ OH) to the stirred solution and stir at 60 ° C for 30 minutes. (1500 rpm) Determine whether the size of the iron oxide nanoparticles varies with the amount of ammonium hydroxide added. 0.6 ml was added to the stirred solution.

3) After the stirring, the prepared aqueous iron oxide solution was cooled at room temperature and stored in a vial.

Example 3: Preparation of hyaluronic acid-iron oxide nanoparticle solution 1

1) Add 5 mL of the iron oxide aqueous solution prepared in Example 2, 250 μL of Span-80, 25 mg of ADH, and 50 mg of hyaluronic acid to 20 mL of paraffin oil in a three-necked flask. Lt; / RTI > (1500 rpm)

2) After adjusting to pH 4.8, 0.1 N HCl was added to the slightly stirred solution.

3) EDC (25 mg) was added to the acid-adjusted stirring solution and stirred for 24 hours.

4) The stirred solution was washed three times with isopropyl alcohol (IPA) and then three times with DW.

Example 4: Preparation of hyaluronic acid-iron oxide nanoparticle solution 2

1) In the same manner as in Example 3, 500 μl of the amount of Span-80, a surfactant, was added.

2) to 4) The same procedure as in Example 3 was carried out.

Experimental Result 1: Measurement of Size of Iron Oxide Nanoparticles

The size of the iron oxide nanoparticles prepared in Example 2 was measured, and the data thereof is shown in FIG. As a result, the size of the iron oxide nanoparticles obtained by adding 0.6 ml of ammonium hydroxide was measured as 39.89 nm. The size of the iron oxide nanoparticles obtained by adding 0.7 ml of ammonium hydroxide was measured to be 40.22 nm. There was a slight difference according to the amount of ammonium hydroxide, but the size was confirmed to be about 40 nm. The condition that the shape of the peak appeared more uniformly was confirmed to be 0.7 ml, and the addition amount of ammonium hydroxide was determined to be 0.7 ml.

Experimental Result 2: Hyaluronic acid-iron oxide nanoparticle spherical composite SEM image

SEM (Scanning Electron Microscope) images of the hyaluronic acid-iron oxide nanoparticle spherical composite prepared in this Example were confirmed and shown in FIGS. 5 to 6. In addition, the above measurement was performed simultaneously using an energy dispersive spectroscope (EDS), and the amount of the elements constituting the hyaluronic acid-iron oxide spherical composite was measured and shown in FIG. As a result, it was confirmed that the weight percentage of Fe was 11.8%, and it was confirmed that Fe ion was contained in the hyaluronic acid spherical composite.

Experimental Result 3: Hyaluronic acid-iron oxide spherical complex fluorescence microscope image

The fluorescence microscope images of the spherical hyaluronic acid-iron oxide complexes prepared in Example 3 and Example 4 were examined and the results are shown in FIGS. 8 and 9. In the fluorescence image of Example 3 prepared by adding 250 [mu] l of Span-80, a surfactant, the shape of the complex was not rounded and uniformity was not uniform. On the other hand, in the fluorescence image of Example 4 prepared by adding 500 ㎕ of Span-80, the shape of the complex was confirmed to be a uniform sphere shape.

As a result, it was confirmed that the shape can be controlled by controlling the amount of the surfactant. It is important to control the amount of surfactant appropriately because a uniform sphere can be used as an MRI contrast agent and a drug delivery system.

The present invention has been described based on the embodiments. It is to be understood by those skilled in the art that the foregoing embodiments are merely illustrative and that various modifications are possible and that these modifications are also within the scope of the present invention.

Claims (20)

(a) preparing a mixed solution by adding an aqueous metal oxide nanoparticle solution, hyaluronic acid, a crosslinking agent and a surfactant to an oil
(b) adding an acid to the mixed solution, and then adding and stirring a coupling agent.
The method according to claim 1,
Wherein the aqueous solution of the metal oxide nanoparticles is an aqueous solution containing iron oxide, manganese oxide, chromium oxide, zinc oxide, nickel oxide, cobalt oxide or gadolinium oxide nanoparticles.
The method according to claim 1,
Wherein the metal oxide nanoparticles have an average diameter of 30 to 50 nm. ≪ RTI ID = 0.0 > 21. < / RTI >
The method according to claim 1,
Wherein the hyaluronic acid-nanoparticle spherical composite is characterized in that a plurality of single metal oxide nanoparticles are inserted and fixed in a spherical hyaluronic acid.
3. The method of claim 2,
The aqueous solution containing the iron oxide nanoparticles
(i) the step of using a ₂ · H ₂ O and FeCl ₃ · H ₂ O was added to FeCl water heating and stirring; And
(ii) adding NH ₄ OH to the stirred aqueous solution, followed by heating and stirring;
Wherein the hyaluronic acid-nanoparticle spherical composite is prepared by a method comprising the steps of:
The method according to claim 1,
The crosslinking agent may be selected from the group consisting of adipic acid dihydrazide (ADH), hexamethylene diamine, cystamine, succinic dihydrazide, hexamethylene diamine, Isophthalic-dihydrazide or malonic dihydrazide, which is characterized in that it is an isophthalic-dihydrazide or malonic dihydrazide.
The method of claim 1, wherein the surfactant is Span-80.
2. The method of claim 1, wherein the coupling agent is EDC.
The method of producing a hyaluronic acid-nanoparticle spherical composite according to claim 1, wherein the hyaluronic acid-nanoparticle spherical composite has an average diameter of 100 to 1000 탆.
A hyaluronic acid-nanoparticle spherical composite wherein the hyaluronic acid polymer produced by the manufacturing method of claim 1 contains a plurality of metal oxide nanoparticles in a spherical hyaluronic acid spherical body.
delete delete delete delete delete An MRI contrast agent comprising as an active ingredient an hyaluronic acid-nanoparticle spherical complex comprising the hyaluronic acid-nanoparticle spherical complex of claim 10.
delete delete A drug delivery system comprising the hyaluronic acid-nanoparticle spherical composite of claim 10 and a pharmaceutically active drug.
delete

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