KR102075823B1 - Composition for hepatic arterial embolization comprising Nanoparticles for genes and anticancer drug delivery - Google Patents

Abstract

The present invention relates to a nanoparticle and a composition comprising the same, which can be used for chemoembolization and simultaneously perform gene therapy and drug treatment, including nanoparticles and embolic materials loaded with an anticancer agent on a siRNA hydrogel capable of gene therapy. .
Since the siRNA hydrogel core carrying the drug of the present invention, the chitosan intermediate layer of the cation surrounding it, and the hyaluronic acid outer layer of the anion are provided, the siRNA and the drug can be released over a long period of time, and thus the drug efficacy is very long.
The composition for chemical embolization comprising the nanoparticles of the present invention can effectively suppress the expression of the target gene together with chemical embolization and drug treatment, thereby increasing the anticancer therapeutic effect.
Nanoparticles of the present invention can be effectively delivered into the cell through the CD44 receptor expressed in liver cancer cells by coating with a hyaluronic acid having a (-) charge can maximize the effect of gene therapy and chemotherapy.

Description

Composition for hepatic arterial embolization comprising nanoparticles for genes and anticancer drug delivery}

The present invention relates to a composition for hepatic artery chemoembolization comprising nanoparticles for chemotherapy and gene therapy. More specifically, the nanoparticles containing an anticancer agent on an siRNA hydrogel capable of gene therapy include embolic material for chemoembolization. The present invention relates to nanoparticles and compositions comprising the same that can be used simultaneously with gene therapy and drug therapy.

Recent advances in imaging technology have made it possible to find the exact location of cancers hidden in the body and to remove them in various ways, including irradiation and endoscopic surgery. However, even if the position of the cancer is correctly developed, surgical removal may not be possible due to various reasons, such as when the cancer is spread to all organs or attached to other organs. In addition, even liver cancer and pancreatic cancer is often found that surgical cure is impossible.

Currently, chemoembolization, the most frequently used procedure for the treatment of liver tumors, is to find arteries that supply liver tumors, administer anticancer drugs, and block the vessels. Liver tissue is supplied with oxygen and nutrients through the portal vein exiting the small and large intestines, and through the hepatic arteries directly from the aorta, with normal liver tissue mainly from the portal vein and tumor tissue mainly from the hepatic artery. Therefore, administering an anticancer agent to the hepatic artery that nourishes the tumor and blocking the blood vessel can selectively necrosis the tumor without harming normal liver tissue. These therapies are the most contributing method for improving liver cancer treatment rate because of the many advantages such as the wide range of application without limitation of the progression of cancer, the limitation of treatment targets. Chemoembolization first involves catheter insertion into the femoral artery located in the inguinal area to access the hepatic artery, and then injected with angiography to obtain the necessary information such as the location, size and blood supply of the tumor. The thin tube is inserted into the catheter to find and target the artery.

The embolic and anticancer mixtures injected for hepatic arterial chemoembolization are emulsions consisting of embolic and anticancer agents, and emulsification (Lipiodol) and adriamycin, an anticancer agent, are mixed in a 1: 4 ratio. commonly used in emulsion form. At this time, adriamycin is used by dissolving in pamiray, an Xray contrast agent similar in specific gravity to Lipiodol, to maintain an emulsion form for a long time. Liver cancer treatment effect by this mixture solution is embolized by Lipiodol embolism and the anticancer agent is delivered to liver cancer, blocking the nutrient supply and maximizing the anticancer effect.

However, the technique was embolized, but the embolic material was washed after a certain time, the drug spreads systemically along the blood vessels can kill normal cells, while there was a limit to kill cancer cells.

The present invention provides nanoparticles and compositions that can effectively infiltrate drugs into cancer cells and provide drug release effects continuously for a long time.

The present invention provides nanoparticles and compositions that can be combined with chemical embolization and anticancer agents as well as gene therapy.

One aspect of the present invention

It relates to a composition for hepatic artery embolization comprising nanoparticles for chemotherapy and gene therapy, an aqueous contrast agent and an embolic material.

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

The nanoparticles for chemotherapy and gene therapy may comprise a siRNA hydrogel carrying a second anticancer agent, chitosan or polyethyleneimine coated on the outer surface of the hydrogel, and a hyaluronic acid outer layer surrounding the chitosan.

In another aspect the invention

Dispersing nanoparticles for chemo and gene therapy in distilled water;

Mixing the dispersion solution with an aqueous contrast agent; And

It relates to a method for preparing a composition for hepatic artery embolization comprising the step of forming an emulsion by adding an embolic material, an oily contrast agent, to a mixed solution containing the aqueous contrast agent.

The method of preparing the composition may form an emulsion by dispersing or mixing the first anticancer agent in the embolic material and then putting it in the mixed solution.

The present invention can include a siRNA nanoparticle for gene therapy in which a second anticancer agent is supported in a conventional chemoembolization formulation, so that the expression of a target gene can be effectively suppressed along with chemoembolization and drug treatment, thereby increasing the anticancer therapeutic effect. In addition, the composition of the present invention includes not only the second anticancer agent supported on the siRNA nanoparticles but also the free first anticancer agent contained in the emulsion composition, so that the first anticancer agent is released and the second anticancer agent is accompanied with the degradation of the siRNA nanoparticles. Since the drug can be released over a long period, chemotherapy can be efficiently performed.

Nanoparticles of the present invention can be effectively delivered into the cell through the CD44 receptor expressed in liver cancer cells by coating with hyaluronic acid can maximize the effect of gene therapy and chemotherapy.

1 is a conceptual diagram of a hepatic artery embolization composition comprising nanoparticles for chemotherapy and gene therapy according to one embodiment of the present invention.
Figure 2 shows an SEM image of anti-cancer-siRNA nanoparticles coated on an siRNA hydrogel with an acrylate chitosan intermediate layer and a hyaluronic acid outer layer.
3 shows a DIC image and a fluorescence image showing that an emulsion was formed by mixing free anticancer agent (doxorubison) and anticancer-siRNA hydrogel-based nanoparticles dissolved in pamiray with Lipiodol.
Figure 4 shows the size of the finished nanoparticles by coating the outer layer with hyaluronic acid after coating the intermediate layer with acrylate chitosan or branched PEI on the siRNA hydrogel, respectively.
FIG. 5 selectively coats the intermediate layer with chitosan or acrylate chitosan and then coats the outer layer with hyaluronic acid to selectively inhibit expression of VEGF gene targeted by nanoparticles delivered to cells under in vitro conditions of the completed nanoparticles. PCR data showing.
Figure 6 shows that the nanoparticles delivered to cells under in vitro conditions of the finished nanoparticles by coating the outer layer with branched PEI and then the outermost layer with hyaluronic acid selectively inhibit the expression of the target VEGF gene. PCR data.
Figure 7 is the result of the cytotoxicity test by the anticancer agent continuously released from the produced anticancer-siRNA nanoparticles.

The present invention can all be achieved by the following description. The following description is to be understood as describing preferred embodiments of the invention, but the invention is not necessarily limited thereto.

Figure 1 shows a conceptual diagram of a composition for hepatic artery (chemical) embolization including nanoparticles for chemo 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 chemotherapy and gene therapy, an aqueous contrast agent 30, and an embolic material 20.

The embolic material 20 is an oily contrast agent, and the nanoparticles 10 are dispersed in an aqueous contrast agent 30. The composition is mixed with an oily contrast agent and an aqueous contrast agent to form an emulsion of water (aqueous) -in-oil (oily) form.

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

The embolic material may be Lipiodol, collagen, thrombin, gelatin, alginic acid, alginate, cellulose acetate, polyvinyl acetate, polyethylenevinyl alcohol, ethylene-vinyl alcohol copolymer, polyvinyl alcohol or mixtures thereof.

The composition may include 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 agent is Iopamidol (Iopamidol), Pamiray (Pamiray), Metrizamide, Ditrizoate (Diatrizoate), Ioxaglate (Ioxaglate), Iopentol, Iomeprol (Iomeprol) ), Group of Iotrolan, Iohexol, Ioversol, Ioxilan, Iopromide, Iodixanol and Iobitridol Can be selected from.

The aqueous contrast agent 30 may disperse the nanoparticles 10 and the first anticancer agent 40.

The anti-cancer treatment and gene therapy nanoparticle 10 is a siRNA hydrogel (11) carrying a second anticancer agent (14), chitosan (12) or polyethyleneimine coated on the outer surface of the hydrogel, and the chitosan or polyethyleneimine It may include a hyaluronic acid outer layer 13 surrounding the.

The siRNA hydrogel 11 targets mRNAs of specific genes. siRNAs can inhibit the expression of certain proteins by binding to messenger RNAs (RNAs) that have the same bases as their sensor strands and break down mRNAs bound to RNA-induced silencing complexes (RISCs). Can be.

The specific gene may be any mRNA that causes cancer or a specific disease. For example, the siRNA hydrogel 11 may be an anti-VEGF siRNA hydrogel that targets vascular endothelial growth factor (VEGF) that causes cancer when excessively expressed.

The siRNA hydrogel may have a molecular weight of 1 × 10 ⁵ g / mol or more. The siRNA hydrogel may be loaded with a small interfering RNA (siRNA) unit sequence of 1 × 10 ³ or more, and may be replicated and loaded in a range of 1 × 10 ³ to 1 × 10 ¹⁰ .

The second anticancer agent 14 may be supported between the double helixes of the siRNA.

The second anticancer agent 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.

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

For example, firstly, the anti-VEGF siRNA hydrogel is dissolved in a solvent (Nuclease free water) at a desired concentration and bath sonicated for about 1 minute, and then a second anticancer drug (eg, doxorubicin, DOX) is loaded. Mix as much as you can and store in the dark for a day. Subsequently, when it is confirmed that the particles loaded with the second anticancer agent in red at the bottom sink, the supernatant is removed by centrifugation (13,200 rpm, 10 minutes) to obtain a siRNA hydrogel loaded with the drug. Supernatants can be stored separately to determine the amount of DOX loaded.

The nanoparticle of the present invention uses chitosan or a cationic polymer as the intermediate layer 12. Preferably the chitosan may be an acrylate chitosan.

The cationic polymer may be any one or more 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 and thus used as intermediate layers coated on the negatively charged siRNA hydrogels and hyaluronic acid (HA). The chitosan and bPEI can reduce the particle size of the hydrogel by condensing or compressing the negatively charged siRNA hydrogel. In addition, the chitosan and bPEI can form a cationic polymer layer having a strong (+) charge on the outer surface of the hydrogel, which is a core, and thus, the hyaluronic acid having a (-) charge is combined with a strong electrostatic attraction to surround the hyaluronic acid. The coating structure can be firmly maintained.

In addition, the chitosan is useful as a drug delivery material in the human body because it forms a structure very similar to our human tissue and excellent human affinity does not cause an immune response.

Because chitosan only dissolves in acid, it does not dissolve well in neutral conditions in the body, which can interfere with the release of anticancer drugs or siRNA hydrogels in the core. In order to solve this problem, in the present invention, the surface functional group of chitosan was modified with acrylate so that chitosan was well decomposed in neutral water.

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

The outer layer of hyaluronic acid exhibits a (-) charge, and therefore, the particles containing the core of the present invention also exhibit a (-) charge on the outside thereof.

Various types of hyaluronic acid are possible for the purposes of the present invention. In particular, a large molecular weight and a small hyaluronic acid can be used, preferably 5-10K, more preferably 5K hyaluronic acid can be used.

      Hyaluronic acid (HA) is a large complex oligosaccharide consisting of up to 50,000 pairs of disaccharide glucuronic acid-β (1-3) N-acetylglucose-amineβ (1-4) as the 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 about 50 nm in diameter.

Nanoparticles of the present invention has the advantage that can be effectively delivered to the cell through the CD44 receptor expressed in liver cancer cells by coating with a hyaluronic acid having a (-) charge to maximize the gene therapy and chemotherapy effect.

The method of the acrylated chitosan or bPEI, hyaluronic acid compared to siRNA hydrogel when the RNA concentration of the siRNA hydrogel aqueous solution is about 100nmole 1: 0.1 ~ 1: 0.5 ~ 2, preferably 1: 0.1 ~ 0.5: 1 ˜2 (hydrogel: chitosan / bPEI: hyaluronic acid) may be added.

The nanoparticles for anticancer treatment 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 of preparing a composition for hepatic artery embolization.

The composition manufacturing method

Dispersing the nanoparticles (10) for chemo and gene therapy in distilled water;

Mixing the dispersion solution with the aqueous contrast agent 20; And

And emulsifying the embolic material 30, which is an oily contrast agent, in a mixed solution containing the aqueous contrast agent to form an emulsion.

The nanoparticles for chemotherapy and gene therapy may use the nanoparticles 10 described above. The nanoparticle 10 is a siRNA hydrogel (11) carrying a second anticancer agent (14), chitosan (12) or cationic polymer coated on the outer surface of the hydrogel and hyaluronic acid surrounding the chitosan or cationic polymer The outer layer 13 may be included.

The method for preparing the nanoparticles 10 includes a step of supporting a second anticancer agent on the siRNA hydrogel, an intermediate layer coating step, a hyaluronic acid coating step, and a separation drying step.

The step of supporting the second anticancer agent on the siRNA hydrogel is a step of mixing the second anticancer agent 14 and the siRNA hydrogel 11 in a solvent (distilled water from which the RNA degrading enzyme activity is removed), and then storing them for a predetermined time and siRNA. If the hydrogel is precipitated may comprise the step of separating from the solvent.

The intermediate layer coating step is a step of coating a mixture of chitosan or cationic polymer in the siRNA hydrogel solution carrying the second anticancer agent.

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

The separation drying step is a step of drying by separating the outer layer coated nanoparticles from the solution.

The method is a weight ratio of the acrylated chitosan, hyaluronic acid relative to the siRNA hydrogel 1: 0.1 to 1: 0.5 to 2, preferably 1: 0.1 to 0.5: 1-2 (hydrogel: chitosan / bPEI: hyaluronic acid) Lonic acid) may be added.

The method may form the emulsion by dispersing or mixing the first anticancer agent 40 in the aqueous contrast agent 30.

The embolic material 20, the aqueous contrast agent 30, the first anticancer agent 40 and the second anticancer agent 14 may be referred to the above-described contents.

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

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

In the method of preparing the composition of the present invention, the nanoparticles for gene therapy carrying the second anticancer agent are redispersed in distilled water (which is distilled water from which RNA degrading enzyme activity is removed) at a desired concentration. Thereafter, the first anticancer agent is dispersed in an aqueous contrast agent (eg, Iopamidol or Pamiray) to a desired concentration, and then mixed with a solution in which the nanoparticles for chemotherapy and gene therapy are redispersed.

At this time, an aqueous contrast agent may be used in a sufficient amount. To mix with the embolic material, the embolic material (eg, Lipiodol) is dispersed in a volume ratio of 1: 1 to 5 (aqueous contrast agent: embolic material) in an aqueous contrast medium solution containing nanoparticles and the first anticancer agent (DOX).

Hereinafter, preferred examples are provided to aid in understanding the present invention, but the following examples are provided only for easier understanding of the present invention, and the present invention is not limited thereto.

Example  One

anti- VEGF siRNA Hydrogel  synthesis

The sense of rat vascular endothelial growth factoer A (VEGF-A) 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.

Ligase-type circular DNA templates were incubated in a reaction buffer (8mM Tris-HCl, 0.4mM spermidine, 1.2mM MgCl ₂ , and 2mM dithiothreitol) for 20 hours at 37 ° C as T7 RNA polymerase. The resulting solution was pipetted several times and sonicated for 5 minutes to decompose the particles. The solution was centrifuged at 13,200 rpm for 20 minutes at 4 ° C. and the supernatant was removed. RNase-free water was then added to wash the particles. The solution was sonicated again 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). For core fabrication, see the prior paper of the present inventors. (Self-assembled RNA interference microsponges for efficient siRNA delivery (nature material, published Nov. 26, 2012))

Doxorubicin (DOX) Supported siRNA Hydrogel  ( DOX - siRNA Hydrogel ) synthesis

Anti-VEGF siRNA hydrogel was mixed with DOX and stored overnight in the dark. Once the bottom loaded DOX loaded particles were found to have settled, they were centrifuged at 13,200 rpm for 10 minutes. The supernatant was walked separately to measure the loadiing amount of DOX to obtain DOX-siRNA hydrogel particles from which the supernatant was removed.

Anti-VEGF siRNA hydrogel was mixed with DOX and stored overnight in the dark. When the particles loaded with DOX in red on the bottom were found to have disappeared, they were centrifuged at 13,200 rpm for 10 minutes. The supernatant was collected separately to measure the loading amount of DOX and DOX-siVEGF hydrogel particles obtained by removing the supernatant were obtained.

Acrylate  Chitosan synthesis

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

Nanoparticle Manufacturing

The 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

(2) 3 μl of sonicated DOX-siRNA hydrogel was mixed with 12 μl of nuclease free water, and then both solutions (1, 2) were mixed.

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

The SEM photograph of the finally prepared nanoparticles is shown in FIG. 2 and the size of the nanoparticles is shown in FIG. 3.

Figure 3 is the size of the nanoparticles coated with chitosan and bPEI, respectively, the intermediate layer, the average size of the particles coated with chitosan intermediate layer is 386.6nm and the average size of the particles coated with bPEI is 362.5nm. The zeta potential of both particles appeared to be minus 10.

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

Referring to Figure 4 it can be seen that the emulsion containing the DOX and siRNA nanoparticles on the aqueous solution.

The left picture of Figure 4 is a micromicrograph showing the formation of the emulsion, the middle picture is a DOX fluorescence micrograph showing the presence of DOX contained in the emulsion and the right picture shows the siRNA hydrogel contained in the emulsion Fluorescence micrographs of siRNA hydrogels. Since DOX fluoresces itself, it did not fluoresce, and siRNA hydrogels fluoresced by chelating a fluorescent substance called YOPRO-1. That is, referring to the photograph of FIG. 4, it can be seen that the emulsion was formed in the solution and that the DOX and the siRNA hydrogel were included in the emulsion.

Gene silencing test method and RT- PCR  data

In FIG. 5, 400,000 HepG2 cells (Korea Cell Line Bank) were seeded in 6 well cell culture plates (cell culture plates) and incubated for one day. After 3 hours, the opti-MEM was removed and washed with DPBS. Then, the opti-MEM mixed with nanoparticles was carefully added, and after 12 hours, the normal meida was changed. After incubation for 24 hours, cells were extracted, RNA was extracted, cDNA was prepared, and gel images were confirmed by PCR and electrophoresis. The cell was treated with control (1), siVEGF hydrogel + 1 mg / ml chitosan + HA (2), siVEGF hydrogel + 1 mg / mL acylated chitosan + HA (3) mixed with medium.

Referring to FIG. 5, it can be seen that the expression of VEGF gene in cells treated with nanoparticles coated with chitosan (2) or acrylated chitosan (3) was significantly reduced compared to control (1). In addition, VEGF gene expression is more suppressed when coated with acrylated chitosan than when coated with normal chitosan.

       In the case of FIG. 6, B16F10 cells were used and proceeded in the same manner as in the above experiment, except that the intermediate layer of the treated nanoparticles was coated with bPEI (1 mg / mL bPEI was used). At this time, it can be seen that the expression of the VEGF gene in the cells treated with the nanoparticles coated with bPEI (2,3) was significantly reduced compared to control (1).

Cell viability test

After 24 hours of 1 × 10 ⁴ seeding of B16F10 cancer cells in a 96 well plate, the control group was divided into groups treated with free DOX (anticancer 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 concentrations and then 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 group, and after 72 hours, the killing effect of 52% was shown.

All simple modifications and variations of the present invention fall within the scope of the present invention, and the specific scope of the present invention will be apparent from the appended claims.

Claims (21)

As a composition comprising a nanoparticle (10), an aqueous contrast agent (30), an embolic material (20) and the first anticancer agent (40) for chemotherapy and gene therapy,
The embolic material is Lipiodol,
The aqueous contrast agent is Iopamidol (Iopamidol), Pamiray (Pamiray), Metrizamide, Ditrizoate (Diatrizoate), Ioxaglate (Ioxaglate), Iopentol, Iomeprol (Iomeprol) ), Group of Iotrolan, Iohexol, Ioversol, Ioxilan, Iopromide, Iodixanol and Iobitridol At least one selected from
The composition forms an emulsion by mixing the aqueous contrast agent and the embolic material in a volume ratio of 1: 1 to 4,
The anticancer and gene therapy nanoparticles and the first anticancer agent are dispersed or dissolved in an aqueous contrast agent,
The anti-cancer and gene therapy nanoparticles
SiRNA hydrogel carrying a second anticancer agent;
Chitosan or polyethyleneimine coated on the outer surface of the hydrogel; And
A hyaluronic acid outer layer surrounding the chitosan or polyethyleneimine,
The nanoparticles for chemotherapy and gene therapy have a size of 100 ~ 400nm, the siRNA hydrogel is loaded with a small interference RNA (siRNA) unit sequence is replicated in the range of 1 × 10 ³ ~ 1 × 10 ¹⁰ , 2 anticancer agent is a composition for hepatic artery embolization, characterized in that carried between the double helix of the siRNA. delete delete delete The composition of claim 1, wherein the composition comprises the nanoparticle and the embolic material in a volume ratio of 1: 2-5. delete delete The composition for hepatic artery embolization according to claim 1, wherein the chitosan is acrylate chitosan. delete delete According to claim 1, wherein the second anticancer agent is doxorubicin (DOX), mitomycin, cisplatin, adriamycin, gemcitabine composition for hepatic artery embolization, characterized in that. delete delete Dispersing nanoparticles for chemo and gene therapy in distilled water;
Mixing the dispersion solution and the first anticancer agent with an aqueous contrast agent; And
A method of preparing a composition for hepatic artery embolization, comprising the step of forming an emulsion by adding an embolic material to a mixed solution containing the aqueous contrast agent,
The method mixes the aqueous contrast agent and the embolic material in a volume ratio of 1: 1 to 4,
The embolic material is Lipiodol,
The aqueous contrast agent is Iopamidol (Iopamidol), Pamiray (Pamiray), Metrizamide, Ditrizoate (Diatrizoate), Ioxaglate (Ioxaglate), Iopentol, Iomeprol (Iomeprol) ), Group of Iotrolan, Iohexol, Ioversol, Ioxilan, Iopromide, Iodixanol and Iobitridol At least one selected from
The anti-cancer and gene therapy nanoparticles
SiRNA hydrogel carrying a second anticancer agent;
Chitosan or polyethyleneimine coated on the outer surface of the hydrogel; And
A hyaluronic acid outer layer surrounding the chitosan or polyethyleneimine,
The anti-cancer and gene therapy nanoparticles are 100 ~ 400nm in size,
The siRNA hydrogel has a small interfering RNA (siRNA) unit sequence in the range of 1 × 10 ³ to 1 × 10 ¹⁰ , wherein the second anticancer agent is supported between the double helices of the siRNA. Manufacturing method. delete 15. The method of claim 14, wherein the method comprises mixing the nanoparticles with the embolic material in a volume ratio of 1: 2-5. delete delete The method of claim 14, wherein the anti-cancer and gene therapy nanoparticles are prepared.
Mixing the second anticancer agent with the siRNA hydrogel in a solvent, storing for a predetermined time, and separating the second anticancer agent from the solvent when the siRNA hydrogel is precipitated;
Mixing and coating acrylated chitosan or polyethyleneimine on the siRNA hydrogel solution carrying a second anticancer agent; And
Mixing hyaluronic acid with the solution to coat hyaluronic acid on the chitosan or polyethyleneimine surface; And
A method for preparing a composition for hepatic artery embolization comprising separating and drying the particles, wherein the method comprises acrylated chitosan and hyaluronic acid in a weight ratio of 1: 0.1 to 1: 0.5 to 2 (hydrogel: Chitosan: hyaluronic acid) method for producing a composition for hepatic artery embolization characterized in that it is added. delete delete

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