WO2019074258A1 - Composition for transcatheter arterial chemoembolization, comprising nanoparticle for anticancer therapy and gene therapy - Google Patents

Abstract

The present invention relates to a nanoparticle that comprises a gene therapy-enabling siRNA hydrogel containing an anticancer agent-impregnated nanoparticle and an embolic material therein and which can be used in chemoembolization and perform both gene therapy and drug therapy, and a composition comprising the same. Designed to comprise an siRNA hydrogel core containing a drug therein, a medium layer of cationic chitosan surrounding the core, and an outer layer of anionic hyaluronic acid, the present invention can release siRNA and the drug for a long period of time and thus shows very long sustainability of a medicinal effect. A composition for transcatheter arterial chemoembolization, comprising the nanoparticle according to the present invention can exert chemoembolization and drug therapy and effectively regulate the expression of a target gene, thereby increasing anticancer therapeutic effect. The nanoparticle of the present invention is coated with hyaluronic acid having (-) charges and thus can be effectively delivered into cells through the CD44 receptor expressed on liver cancer cells, bringing about a maximum effect on gene therapy and anticancer therapy.

Description

Composition for hepatic artery chemoembolization including nanoparticles for chemotherapy and gene therapy

The present invention relates to a composition for hepatic artery chemoembolization comprising nanoparticles for chemotherapy and gene therapy. More particularly, the present invention relates to a composition for hepatocarcinoma chemoembolization comprising gene therapy capable of inhibiting nanoparticles containing chemotherapeutic agents, And to a nanoparticle and a composition containing the nanoparticle which can simultaneously perform gene therapy and drug treatment.

Recent advances in imaging technology have led to the discovery of the exact location of the cancer in the body, which can then be removed by a variety of methods, including radiation and endoscopic surgery. However, even if the position of the cancer is accurately developed, surgical removal can not be performed due to various reasons such as the cancer spreading to the whole organs or the other organ. In addition, liver cancer and pancreatic cancer, even if found in the surgical cure is often impossible.

Currently, chemoembolization is the treatment most commonly used for the treatment of liver tumors. It is a treatment to block blood vessels by administering anticancer drugs to find arteries supplying nutrients to liver tumors. The liver tissue is supplied with oxygen and nutrients through the portal vein and the hepatic artery directly from the small intestine and the large intestine. Normal liver tissue is mainly supplied in the context, and tumor tissue is supplied mainly from the hepatic artery. Therefore, administering anticancer drugs to the hepatic artery supplying the tumor and blocking the blood vessels can selectively necrotize the tumor without harming the normal liver tissue. These treatments are the most important method to improve the treatment rate of liver cancer because there are no limitations on the degree of cancer progression, the scope of application is wide, In chemoembolization, the catheter is first inserted into the femoral artery in the inguinal region, and after approaching the hepatic artery, the angiographic agent is injected to obtain information necessary for the treatment, such as the location, size and blood supply pattern of the tumor. Is inserted into the catheter to find the target artery.

The injected embolization substance and the anticancer drug mixture for embolization of hepatic arterial chemoembolization are emulsion composed of an embolization substance and an anticancer drug and mixed with 1: 4 ratio of an embolizing substance Lipiodol and an anticancer drug adriamycin in a ratio of 1: emulsion. At this time, adriamycin is dissolved in the pamirite, an Xray contrast agent having a specific gravity similar to that of lipiodol, so that it can be maintained in a long-time emulsion form. The effect of this mixture on hepatocarcinoma treatment is due to the transfer of anticancer drug to hepatocellular carcinoma along with the embolization effect of Lipiodol, thereby preventing the supply of nutrients and maximizing the anticancer effect.

However, although the embolization technique was performed, the embolization material was washed after a certain time, and the drug spreads throughout the blood vessels to kill the normal cells, but it was not enough to kill the cancer cells.

The present invention provides nanoparticles and compositions that can effectively penetrate a drug into cancer cells and at the same time provide sustained drug release effects.

The present invention is to provide nanoparticles and compositions that can be used in combination with chemotherapeutic agents and anticancer agents as well as gene therapy.

One aspect of the present invention is

A composition for hepatic artery embolization comprising nanoparticles for cancer therapy and gene therapy, a water contrast agent and an embolus substance.

The composition may further comprise a first anticancer agent dispersed or dissolved in the aqueous contrast medium or the embolic material.

The nanoparticles for chemotherapy and gene therapy may include a second anti-cancer agent-carrying siRNA hydrogel, chitosan or polyethylene imine coated on the outer surface of the hydrogel, and a hyaluronic acid outer layer surrounding the chitosan.

In another aspect,

Dispersing nanoparticles for chemotherapy and gene therapy in distilled water;

Mixing the dispersion solution with a water-based contrast agent; And

And injecting an embolic agent, which is an oil contrast agent, into a mixed solution containing the aqueous contrast agent to form an emulsion.

The composition may be prepared by dispersing or mixing the first anticancer agent in the embolization agent, and then adding the first anticancer agent to the mixed solution to form an emulsion.

Since the present invention includes siRNA nanoparticles for gene therapy in which the second chemotherapeutic agent is carried in a conventional chemoembolization agent, the expression of the target gene can be efficiently inhibited together with the chemoembolization and drug treatment, thereby enhancing the effect of the chemotherapeutic treatment. In addition, the composition of the present invention contains the first anticancer drug contained in the emulsion composition as well as the second anticancer drug loaded on the siRNA nanoparticles, and thus the first anticancer drug is initially released and the second anticancer drug is degraded with the siRNA nanoparticle decomposition Since the drug can be released over a long period of time, the chemotherapy can be efficiently performed.

By coating the nanoparticles of the present invention with hyaluronic acid, the CD44 receptor expressed in hepatocarcinoma cells can be effectively delivered into the cells, thereby maximizing the gene therapy and the anti-cancer therapeutic effect.

1 is a conceptual diagram of a hepatic artery chemoembolization composition including nanoparticles for chemotherapy and gene therapy according to an embodiment of the present invention.

FIG. 2 shows an SEM image of the anti-cancer-siRNA nanoparticles coated with the acrylate chitosan intermediate layer and the hyaluronic acid outer layer on the siRNA hydrogel.

FIG. 3 shows a DIC image and a fluorescence image showing that an emulsion is formed by mixing a free anticancer drug (toxin Rubisone) dissolved in Pamirai and anti-cancer-siRNA hydrogel-based nanoparticles with lipiodol.

FIG. 4 shows the size of the completed nanoparticles by coating an intermediate layer with an acrylate chitosan or branched PEI on an siRNA hydrogel and coating the outer layer with hyaluronic acid.

FIG. 5 is a graph showing the results of coating the outer layer with hyaluronic acid after coating the intermediate layer with chitosan or acrylate chitosan and selectively inhibiting the expression of the targeted VEGF gene in the nanoparticles transferred to the cells under the in vitro condition of the finished nanoparticles Lt; / RTI >

FIG. 6 shows that the nanoparticles transferred to the cells under the in vitro condition of the completed nanoparticles selectively coat the targeted VEGF gene expression by coating the intercalation layer with branched PEI and coating the outermost layer with hyaluronic acid PCR data.

FIG. 7 shows the results of cytotoxicity experiments with anticancer drugs continuously released from the prepared anti-cancer-siRNA nanoparticles.

The present invention can be all accomplished by the following description. The following description should be understood to describe preferred embodiments of the present invention, but the present invention is not necessarily limited thereto.

1 is a conceptual diagram of a composition for hepatic artery (chemo) embolization including nanoparticles for chemotherapy and gene therapy according to an embodiment of the present invention.

Referring to FIG. 1, the hepatic artery embolization composition of the present invention includes nanoparticles 10 for cancer therapy and gene therapy, water-based contrast agent 20, and embolic material 30.

The embolic material 30 is an oil-based contrast agent, and the nanoparticles 10 are dispersed in the aqueous contrast medium 20. The composition is prepared by mixing the oil-based contrast agent and the water-based contrast agent to form an emulsion in an oil (oily) -in-water (aqueous) form.

The composition may further comprise a first anticancer agent 40 dispersed or dissolved in the aqueous contrast medium or oily embolic material. That is, both the hydrophilic or hydrophobic anticancer agent may be used as the first anticancer agent.

The embolic material may be lipiodol, collagen, thrombin, gelatin, alginic acid, alginate, cellulose acetate, polyvinyl acetate, polyethylene vinyl alcohol, ethylene-vinyl alcohol copolymer, polyvinyl alcohol or a mixture thereof.

The composition may contain the nanoparticles and the embolic material in a volume ratio of 1: 1 to 5, preferably 1: 1 to 4, more preferably 1: 2 to 5.

The aqueous contrast medium may be selected from the group consisting of Iopamidol, Pamiray, Metrizamide, Diatrizoate, Ioxaglate, Iopentol, Iomeprol ), Iotrolan, Iohexol, Ioversol, Ioxilan, Iopromide, Iodixanol, and Iobitridol. ≪ / RTI >

The aqueous contrast agent 20 may disperse the nanoparticles 10 and the first anticancer agent 40.

The chemotherapeutic and gene therapy nanoparticles 10 may be prepared by mixing an siRNA hydrogel 11 carrying a second anticancer drug 14, chitosan 12 or polyethyleneimine coated on the outer surface of the hydrogel and a chitosan or polyethyleneimine And a hyaluronic acid outer layer 13 surrounding the outer layer 13.

The siRNA hydrogel (11) targets mRNAs of a specific gene. siRNA binds messenger RNA (mRNA), which has the same nucleotide sequence as its sense strand, and degrades the expression of a specific protein by degrading mRNA bound to RNA induced silencing complex (RISC) .

The specific gene may be cancer or any mRNA which induces a specific disease. For example, the siRNA hydrogel 11 may be an anti-VEGF siRNA hydrogel targeting an endothelial growth factor (VEGF) that causes cancer if overexpressed.

The siRNA hydrogel may have a molecular weight of 1 x 10 < ⁵ > g / mol or more. The siRNA hydrogel may contain at least 1 x 10 ³ small interfering RNA (siRNA) unit sequences, and may be replicated more than 1 x 10 ³ to 1 x 10 ¹⁰ ranges.

The second anticancer agent 14 may be carried between the double helix of the siRNA.

The second anticancer drug 14 may be doxorubicin (DOX), mitomycin, cisplatin, adriamycin.

The first anticancer agent 40 and the second anticancer agent 14 may be the same.

The method of supporting the drug on the siRNA hydrogel is as follows.

Mixing the drug and the siRNA hydrogel in a solvent (water), storing the mixture for a predetermined time, and separating the siRNA hydrogel from the solvent when the hydrogel is precipitated.

For example, if you want to release the anti-VEGF siRNA hydrogel to a desired concentration in a solvent (Nuclease free water), and then apply bath sonication for about 1 minute and then load the second anticancer drug (eg, doxorubicin, DOX) And stored in the dark room for one day. Then, when it is confirmed that the particles loaded with the second anticancer drug on the bottom are settled down, centrifugation (13,200 rpm, 10 minutes) is performed to remove the supernatant, and the drug-loaded siRNA hydrogel can be obtained. The supernatant can be stored separately to determine the amount of DOX loaded.

The nanoparticles of the present invention use chitosan or a cationic polymer as the intermediate layer (12). Preferably, the chitosan may be acrylate chitosan.

The cationic polymer may be at least one selected from the group consisting of polyethyleneimine, polyamine and polyvinylamine, and preferably branched PEI having relatively low toxicity may be used.

In the present invention, the chitosan and bPEI are positively charged, so they are used as an intermediate layer coated with a negatively charged siRNA hydrogel and hyaluronic acid (HA). The chitosan and bPEI can condense or compress the negatively charged siRNA hydrogel to reduce the particle size of the hydrogel. In addition, since the chitosan and bPEI can form a cationic polymer layer having a strong (+) charge on the outer surface of the core hydrogel, it binds to (-) charged hyaluronic acid with strong electrostatic attraction, The coating structure can be firmly maintained.

In addition, the chitosan has a structure very similar to that of human tissues and has excellent affinity for human body, so that the chitosan is useful as a drug delivery material in the human body since an immune response does not occur.

Because chitosan is soluble only in acid, it is not well soluble in neutral conditions in the human body and may interfere with the release of anticancer drugs or siRNA hydrogels in the core. To solve this problem, in the present invention, the surface functional group of chitosan is modified with acrylate so that chitosan is decomposed well in neutral water.

The outer layer 13 is a layer of hyaluronic acid coated on the outer surface of the intermediate layer 12.

The hyaluronic acid outer layer represents a negative charge, and therefore the particles including the core of the present invention also exhibit negative charges.

Various forms of hyaluronic acid are possible for the purposes of the present invention. Particularly, a hyaluronic acid having a large molecular weight and a small hyaluronic acid may be used, preferably 5 to 10 K, more preferably 5 K hyaluronic acid.

      Hyaluronic acid (HA) is a large complex oligosaccharide consisting of up to 50,000 pairs of disaccharide glucuronate-β (1-3) N-acetylglucose-amine β (1-4) as a basic unit. It is found in vivo as a major component of the extracellular matrix. Its tertiary structure is in the form of a random coil with a diameter of about 50 nm.

The nanoparticles of the present invention are effectively delivered to the cells through the CD44 receptor expressed on hepatocarcinoma cells by coating with (-) charged hyaluronic acid, thereby maximizing the effect of gene therapy and chemotherapy.

The method comprises reacting the acrylated chitosan or bPEI and hyaluronic acid with the siRNA hydrogel in an amount of 1: 0.1-1: 0.5-2, preferably 1: 0.1-0.5: 1, when the RNA concentration of the aqueous solution of siRNA hydrogel is about 100 nmole ~ 2 (hydrogel: chitosan / bPEI: hyaluronic acid) may be added.

The nanoparticles for chemotherapy and gene therapy of the present invention may have a size of 100 to 400 nm, preferably 200 nm or less.

In another aspect, the present invention relates to a method for preparing a composition for hepatic artery embolization.

The composition manufacturing method includes

Dispersing nanoparticles (10) for chemotherapy and gene therapy into distilled water;

Mixing the dispersion solution with a water-based contrast agent (20); And

And injecting the embolization material 30, which is an oil contrast agent, into a mixed solution containing the aqueous contrast agent to form an emulsion.

As the nanoparticles for chemotherapy and gene therapy, the nanoparticles 10 described above can be used. The nanoparticles 10 may be formed of an siRNA hydrogel 11 carrying a second anticancer drug 14, chitosan 12 coated on the hydrogel outer surface or a cationic polymer and hyaluronic acid surrounding the chitosan or cationic polymer, And an outer layer 13.

The method for preparing the nanoparticles 10 includes a step of carrying a second anticancer agent on the siRNA hydrogel, an interlayer coating step, a hyaluronic acid coating step, and a separating and drying step.

The step of supporting the second anticancer agent on the siRNA hydrogel comprises: mixing the second anticancer agent (14) and the siRNA hydrogel (11) in a solvent (distilled water from which the RNAase activity is removed) And separating the hydrogel from the solvent when the hydrogel is precipitated.

In the intermediate layer coating step, chitosan or a cationic polymer is mixed with the siRNA hydrogel solution containing the second anticancer drug.

The outer layer coating step includes mixing hyaluronic acid in the solution and coating hyaluronic acid on the surface of the chitosan or cationic polymer.

In the separation and drying step, the nanoparticles coated with the outer layer are separated from the solution and dried.

The method comprises reacting the acrylated chitosan and hyaluronic acid with the siRNA hydrogel in a ratio of 1: 0.1-1: 0.5-2, preferably 1: 0.1-0.5: 1-2 (hydrogel: chitosan / bPEI: hyaluronic acid) Can be added.

In the method, the first anticancer drug 40 may be dispersed or mixed in the embolic material 30, and then the emulsion may be formed by mixing the first anticancer drug 40 and the first anticancer drug 40 in the mixed solution.

The embolization substance 30, the aqueous contrast medium 20, the first anticancer agent 40 and the second anticancer agent 14 may be referred to above.

In the method, the nanoparticles and the embolic material may be mixed at a volume ratio of 1: 1 to 5, preferably 1: 1 to 4 or 1: 2 to 5.

The method can mix the embolic material and the aqueous contrast agent in a volume ratio of 1: 1 to 5, preferably 1: 1 to 4.

In the method for producing a composition of the present invention, the gene therapy nanoparticles carrying the second anticancer drug are redispersed in distilled water (distilled water from which the RNAase activity is removed) to a desired concentration. Thereafter, the first anticancer drug is dispersed in a water-based contrast agent (for example, Iopamidol or Pamiray) so as to have a desired concentration, and then mixed with a redispersed solution of the nanoparticles for chemotherapy and gene therapy.

A sufficient amount of aqueous contrast medium can be used. For mixing with an embolus substance, an embolization substance (for example, lipiodol) is dispersed in a volume ratio (embolization substance: aqueous contrast medium) of 1: 1 to 5 to a water-based contrast agent solution containing nanoparticles and a first anticancer drug (DOX).

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following embodiments are provided for the purpose of easier understanding of the present invention, but the present invention is not limited thereto.

Example 1

anti-VEGF siRNA hydrogel synthesis

The sense of vascular endothelial growth factor (VEGF-A) in rats is 5'-AUGUGAAUGCAGACCAAAGAA TT-3 'and the antisense is 5'-UUCUUUGGUCUGCAUUCA CAU TT-3'. Long linear single stranded DNA encoding the sense and antisense was prepared.

The ligated circular DNA template was incubated with the reaction buffer (8 mM Tris-HCl, 0.4 mM spermidine, 1.2 mM MgCl ₂ , and 2 mM dithiothreitol) at 37 ° C for 20 hours as in T7 RNA polymerase. The resulting solution was pipetted several times and sonicated for 5 minutes to disintegrate the particles. The solution was centrifuged at 13,200 rpm for 20 minutes at 4 DEG C and the supernatant was removed. The particles were then washed with RNase-free water. The solution was again sonicated for 1 minute and centrifuged. The washing step was repeated three more times to remove the RCT reagent to obtain RNA-microsponge (anti-VEGF siRNA hydrogel). The concentration of RNA-microsponge was measured using Quant-iT RNA BR assay kits (Invitrogen). The self-assembled RNA interference microsponges for efficient siRNA delivery (nature material, published on Nov. 26, 2012) were used for core preparation.

Synthesis of siRNA hydrogel (DOX-siRNA hydrogel) carrying Doxorubicin (DOX)

DOX was mixed with anti-VEGF siRNA hydrogel and stored overnight in a dark room. When it was confirmed that the DOX-loaded particles were red in the bottom, they were centrifuged at 13,200 rpm for 10 minutes. The supernatant was taken apart to measure the amount of DOX loadiing and the DOX-siRNA hydrogel particles with the supernatant removed.

DOX was mixed with anti-VEGF siRNA hydrogel and stored overnight in the dark room. When it was confirmed that the DOX-loaded particles in the red color on the bottom were not scattered, they were centrifuged at 13,200 rpm for 10 minutes. The supernatant was taken apart to measure the loading amount of DOX and the DOX-siVEGF hydrogel particles with the supernatant removed.

Acrylate  Chitosan synthesis

80 mg of 48 kDa chitosan was mixed with 8 mL distilled water (DW), and then 182.4 μL of 0.4 M acetic acid was added, and the solution was completely dissolved for 1 to 2 hours while stirring at 60 ° C. 400 μL of 0.05% NaOH and 472 μL of Glycidyl methacrylate (GMA) were added and stirred at 60 ° C. overnight. After the temperature was lowered to room temperature, 5 mL of 2M NaOH and 10 μL of acetic anhydride were added and stirred for 3 to 4 hours. And centrifuged several times for 10 minutes at 4,000 rpm using a dialysis membrane to remove unreacted materials to obtain the following acrylate chitosan.

Nanoparticle manufacturing

The previously prepared acrylate chitosan was dissolved in nuclease free water at 1 mg / mL. 48 kDa chitosan and 25 kDa bPEI were also dissolved in nuclease free water at 1 mg / mL.

(1) 1 μl of 1 mg / mL chitosan / bPEI and 9 μl of nuclease free water were mixed

(2) Add 12 μl of nuclease free water to 3 μl of DOX-siRNA hydrogel sonicated, and mix the two solutions (1 and 2).

The mixed solution was incubated for 10 minutes and sonicated. The mixture was mixed with 5 μl of 1 mg / mL hyaluronic acid (HA) and incubated for 30 minutes.

SEM photographs of the finally prepared nanoparticles are shown in FIG. 2, and the sizes of the nanoparticles are shown in FIG.

FIG. 3 shows the sizes of the nanoparticles coated with chitosan and bPEI as the intermediate layer and 386.6 nm as the average size of particles coated with chitosan as the middle layer, and the average size of particles coated with bPEI was 362.5 nm. The zeta potential of both particles was about minus 10.

Preparation of composition for embolization

The DOX-siRNA hydrogel loaded with the fluorescent dye YOPRO-1 was mixed with bPEI and hyaluronic acid-coated nanoparticles and free DOX dissolved in Pamirai. At this time, the amount of DOX loaded on the nanoparticles and the amount of free DOX were mixed to a total of 6.25 mg / mL. An emulsion containing nanoparticles and free DOX can be obtained by mixing the solution with lipoid in a ratio of 1: 2 using 3 way pumping.

Referring to FIG. 4, it can be seen that an emulsion containing DOX and siRNA nanoparticles is present on the aqueous solution.

FIG. 4 is a micrograph showing the formation of an emulsion. FIG. 4 is a DOX fluorescence microscope photograph showing the presence of DOX contained in the emulsion. The right image shows the presence or absence of siRNA hydrogel contained in the emulsion It is a fluorescence microscope photograph of a siRNA hydrogel. DOX did not fluoresce by itself because it was fluorescent, and siRNA hydrogel chelates YOPRO-1 to confirm fluorescence. That is, referring to the photograph of FIG. 4, it can be confirmed whether or not an emulsion is formed in the solution and that DOX and siRNA hydrogel are contained in the emulsion.

Gene silencing experiment method and RT-PCR data

In case of FIG. 5, 400,000 HepG2 cells (Korean Cell Line Bank) were seeded in a 6 well cell culture plate (cell culture plate) and incubated for one day. After 3 hours, opti-MEM was removed and washed with DPBS. Opti-MEM mixed with nanoparticles was carefully placed and replaced with normal meida after 12 hours. After incubation for 24 hours, the cells were extracted and RNA was extracted and cDNA was obtained. The gel image was confirmed by PCR and electrophoresis. The cells were treated with control (1), siVEGF hydrogel + 1 mg / ml chitosan + HA, siVEGF hydrogel + 1 mg / mL acylated chitosan + HA (3)

5, the expression of the VEGF gene in cells treated with chitosan (2) or acrylated chitosan (3) coated nanoparticles was significantly reduced compared to control (1). In addition, VEGF gene expression is more inhibited when coated with acrylated chitosan than with chitosan.

    In Fig. 6, B16F10 cells were used and the same procedure as in the above experiment was carried out except that the intermediate layer of the treated nanoparticles was coated with bPEI (using 1 mg / mL bPEI). At this time, the expression of the VEGF gene in the cell treated with bPEI (2, 3) -coated nanoparticles was markedly reduced compared to control (1).

Cell viability test

The cells were treated with 1 × 10 ⁴ seeding of B16F10 cancer cells in a 96-well plate, followed by treatment with free DOX (anti-cancer agent) and nanoparticles of Example 1 (DOX-siVEGF + acrylate chitosan + HA) The experiment was carried out.

All three groups were treated with 200 nM DOX concentration and stored for 6 hours. Cell viability is shown in FIG. 7 after 24 hours, 48 hours, and 72 hours after treatment with new cell media after removal of the anticancer drug.

Referring to FIG. 7, after 48 hours, Example 1 showed a killing effect of 23% compared to the control, and after 72 hours, the killing effect was 52% as compared with the control.

It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the appended claims.

 The composition of the present invention can be administered simultaneously with chemoembolization, drug therapy and gene therapy, and thus can be used for the treatment of liver cancer.

Claims (21)

1. Compositions for hepatic artery embolization comprising nanoparticles for cancer therapy and gene therapy, aqueous contrast agents, and embolic agents.
2. The composition for hepatic artery embolization according to claim 1, wherein the embolic material is a contrast agent and the nanoparticles are dispersed in a water-based contrast agent to form an emulsion.
3. The composition for hepatic artery embolization according to claim 1, wherein the composition further comprises a first anti-cancer agent dispersed or dissolved in the aqueous contrast agent or the embolic material.
4. The method of claim 1, wherein the embolic material is selected from the group consisting of lipiodol, collagen, thrombin, gelatin, alginic acid, alginate, cellulose acetate, polyvinyl acetate, polyethylene vinyl alcohol, ethylene-vinyl alcohol copolymer, polyvinyl alcohol, Wherein the composition for hepatic artery embolization comprises:
5. The composition for hepatic artery embolization according to claim 1, wherein the composition comprises the nanoparticles and the embolic material in a volume ratio of 1: 1 to 5: 1.
6. 7. The composition of claim 1, wherein the aqueous contrast agent is selected from the group consisting of Iopamidol, Pamiray, Metrizamide, Diatrizoate, Ioxaglate, Iopentol, , Iomeprol, Iotrolan, Iohexol, Ioversol, Ioxilan, Iopromide, Iodixanol and Iovitol, (Iobitridol). ≪ / RTI > < RTI ID = 0.0 > 11. < / RTI >
7. The method of claim 1, wherein the nanoparticles for chemotherapy and gene therapy comprise

   An siRNA hydrogel carrying a second anticancer drug;

     Chitosan or polyethyleneimine coated on the outer surface of the hydrogel; And

     And a hyaluronic acid outer layer surrounding the chitosan or polyethyleneimine.
8. The composition for hepatic artery embolization according to claim 7, wherein the chitosan is acrylate chitosan.
9. [8] The composition for hepatic artery embolization according to claim 7, wherein the siRNA hydrogel targets mRNA of a specific gene and inhibits its expression.
10. [8] The composition according to claim 7, wherein the second anticancer agent is carried between double strands of the siRNA.
11. 8. The composition for hepatic artery embolization according to claim 7, wherein the second anticancer agent is doxorubicin (DOX), mitomycin, cisplatin, adriamycin, gemcitabine.
12. 8. The method of claim 7, wherein the siRNA hydrogel and the hyaluronic acid layer are (-) charges and the chitosan to polyethylene imine layer is (+) charged, A composition for embolization.
13. [8] The composition for hepatic artery embolization according to claim 7, wherein the nanoparticles for chemotherapy and gene therapy have a size of 100 to 400 nm.
14. Dispersing nanoparticles for chemotherapy and gene therapy in distilled water;

    Mixing the dispersion solution with a water-based contrast agent; And

    Forming an emulsion by adding an embolic agent, which is an oil contrast agent, to a mixed solution containing the aqueous contrast agent.
15. 15. The method according to claim 14, wherein the first anticancer agent is dispersed or mixed in the embolic material, and the emulsion is formed in the mixed solution.
16. 15. The method according to claim 14, wherein the nanoparticles and the embolic material are mixed at a volume ratio of 1: 1 to 5: 5.
17. 15. The method according to claim 14, wherein the embolization substance and the aqueous contrast agent are mixed at a volume ratio of 1: 1 to 4: 1.
18. 15. The method according to claim 14, wherein the nanoparticles for chemotherapy and gene therapy

    An siRNA hydrogel carrying a second anticancer drug;

      Chitosan or polyethyleneimine coated on the outer surface of the hydrogel; And

      And a hyaluronic acid outer layer surrounding the chitosan to polyethyleneimine.
19. 15. The method according to claim 14, wherein the nanoparticles for chemotherapy and gene therapy are prepared

    Supporting the second anticancer agent comprising the second anticancer agent and the siRNA hydrogel in a solvent for storage for a predetermined period of time, and separating the second anticancer agent and the siRNA hydrogel from the solvent when the siRNA hydrogel is precipitated, on the siRNA hydrogel;

    Mixing and coating acrylate chitosan or polyethyleneimine with the siRNA hydrogel solution containing the second anticancer agent; And

    Mixing the solution with hyaluronic acid to coat the surface of the chitosan or polyethylene imine with hyaluronic acid; And

    And separating and gauging the particles. The method for preparing a composition for hepatic artery embolization according to claim 1,
20. 20. The method according to claim 19, wherein the method comprises adding the acrylated chitosan and hyaluronic acid to the siRNA hydrogel in a ratio of 1: 0.1-1: 0.5-2 (hydrogel: chitosan: hyaluronic acid) ≪ / RTI >
21. Carrying an anticancer agent comprising the anticancer agent and the siRNA hydrogel in a solvent, storing the mixture for a predetermined time, and separating the siRNA hydrogel from the solvent when the siRNA hydrogel is precipitated, on the siRNA hydrogel;

    Mixing and coating the acrylated chitosan with the siRNA hydrogel solution carrying the anticancer agent; And

    Mixing the solution with hyaluronic acid to coat the surface of the chitosan with hyaluronic acid; And

    And separating and purifying the particles. The method for producing nanoparticles for chemotherapy and gene therapy.

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