KR20140119358A - Anticancer agent-coated drug eluting covered stent and method thereof - Google Patents

Abstract

The present invention relates to a drug release type covered stent covalently bound with an anticancer agent and to a method for manufacturing the same. With respect to a method for manufacturing a covered stent for digestive organ cancers, the method comprises the steps of: binding allyl alcohol, which is a chemical functional group for drug binding, to a surface of a covered stent, using a plasma reactor; and binding cystamine, which is a chemical functional group for drug binding, to the ally alcohol-bound covered stent, using 1,1-carbonyldiimidazole, thereby producing a cystamine-bound cover stent. The anticancer agent for cancer treatment is chemically coated on the covered stent, thereby providing a stent capable of preventing restenosis after surgery and treating small topical regions.

Description

TECHNICAL FIELD [0001] The present invention relates to a drug-releasing coated stent and an anticancer drug-coated drug-eluting covered stent,

The present invention relates to a covered stent, and more particularly, to a method of manufacturing a drug-released coated stent in which a drug is covalently bonded. The coated stent is plasma-treated with a silicon material using a plasma apparatus, To a drug-releasing coated stent covalently bound with an anticancer agent coated with an anticancer agent and a method for producing the same.

Gastric cancer, such as gastric cancer, esophageal cancer, colon cancer, biliary cancer, pancreatic cancer, is a very common cancer in Korea. When these cancers are found early, surgical resection is found to be the best treatment or inoperable cases in a few cases. Progressive digestive cancer blocks the gastrointestinal tract or bile ducts, causing symptoms such as dysphagia, vomiting, and jaundice. In order to improve patient survival and quality of life, we need to open the closure tube. Endoscopy or radiological intervention stent implantation is recognized as the primary treatment rather than past bypass surgery.

     Generally, a stent is a medical device that is inserted and attached to a vessel or conduit in a constricted body to expand the constriction site back to its original size. Stents used in gastrointestinal cancers are made of metals such as stainless steel, nickel titanium, nitinol, and cobalt chromium and are inserted into the body through a thin diameter emitting device. An enlarged stent is called an expandable stent. The stent is used for non-vascular systems such as airway, airway, and gastrointestinal tract. These stents do not have direct cancer cell killing effect, so they have a disadvantage that restenosis of the stent due to the growth of cancer cells occurs after a certain period of time after the stent is inserted. Therefore, a drug such as an anticancer drug is coated on an existing stent, and the drug is gradually released, so that the function of inhibiting the growth of cancer cells in the place where the stent is inserted is needed. The development of drug eluting stents (DES), a concept of local drug delivery, is an urgent need in stents for digestive cancer.

The drug-eluting stent is usually divided into a bare metal stent or a covered stent coated with non-degradable polymer. The drug-releasing drug is coated on the surface of the stent with anticancer drugs to control drug release . There are three main ways to carry drugs on the stent. There is a method of directly attaching a drug itself to a metal stent without a polymer, or a method of loading a drug carrying a polymer in a porous metal stent. Finally, non-degradable and biodegradable polymers can be used as a method of coating the polymer with the drug on the stent. When the drug is dissolved in the non-degradable polymer, the drug is released by diffusion. The drug release tendency is controlled by the thickness of the coated polymer, and the drug releasing stent CYPHER ( ^TM ) or TAXUS ( ^TM ) . When a biodegradable polymer is used, the biodegradation of the drug-bearing polymer occurs over time, and the drug is eluted.

Non-vascular stents are commonly used in the organs, bronchi, esophagus, and feces, and two common problems must be avoided. The first is the proliferation of the stomach, which results in a benign hyperplasia of the stomach resulting in narrowing of the lumen. Second, malignant hyperplasia occurs in the stent inserted to treat obstruction of the body passageway by cancer. The main goal of developing a gastro-intestinal drug-eluting stent is to prevent the overexpression of these tumors. Polyurethane (PU), polyethylene, Teflon, polytetrafluoroethylene (PTFE), polycarprolactone (PCL), polylactic acid-co polyglycolic acid (PLGA), polyvinyl alcohol (PVA) and the like can be used as a medium for carrying a drug, so that a large amount of drug can be coated on the stent.

However, when the degradable polymer is coated on the stent, the surface of the polymer is peeled or destroyed in the course of performing most of the procedures, thereby losing original intrinsic drug release characteristics, injuring the tubular wall, promoting positive stenosis, And the like. On the other hand, the coated stent coated with the non-degradable polymer has a disadvantage in that it is difficult to carry the drug during processing and the release of the drug is not controlled uniformly. In addition, when the drug is loaded in the process of coating the non-degradable polymer on the covered stent, the chemical structure of the drug is destroyed and the anticancer activity is lost because the drug is carried out at high temperature and high pressure.

In view of these circumstances, there is a need for a coated stent coating technique that allows the drug to be constantly released from the digestive tract while maintaining the anticancer activity of the drug.

The object of the present invention is to provide a method of chemically modifying a surface of a covered stent by plasma treatment and coating an anticancer agent on a surface of a covered stent, And a method for producing the same.

It is an object of the present invention to provide a method for manufacturing a covered stent for a digestive arm, the method comprising the steps of: binding allyl alcohol, which is a chemical functional group for drug binding, to the surface of a covered stent using a plasma reactor; A cystamine-coupled covered stent, which is a chemical reactor for drug-binding, is prepared by binding cystamine to the allyl alcohol-bound covered stent using 1,1-carbonyldiimidazole. The method comprising the steps of: (a) preparing a drug-releasing coated stent;

Preferably, the chemical reactor for the drug-binding of the present invention is an allyl alcohol propoxylate, a glycidol, an allylamine, a butenylamine, an allyl proline, Allyl indole carboxylic acid, allyl oxopyrrolidine carboxylic acid, allyl bromide, allyl cysteine, allyl cystamine, allyl cysteine, ) And acryloyl cystamine, and the linker for binding the drug is at least one selected from the group consisting of cystamine, succinic anhydride, diethylamine ( diethyl amine, hexamethylene diamine, adipic acid dihydrazide, succinic dihydrazide, jade, From the group consisting of Oxalyldihydrazide, Carbohydrazide, Semicarbazide, and Glutamic acid hydrazide, and Hydrazide, And is selected arbitrarily.

More preferably, the linker of the present invention is degraded by glutathione or hydrogen ion concentration.

Preferably, the vacuum of the plasma reactor of the present invention is adjusted to 1 to 100 mHg and the surface of the covered stent is treated with argon plasma for 1 to 5 minutes at a flow rate of 1 to 500 cm < ³ > / m, And the allyl alcohol is bonded to the covered stent by treatment with 1 to 100 W of glow discharge for 1 to 60 minutes.

Another object of the present invention is to provide a method for producing a covered stent for a digestive arm, which comprises dissolving gemcitabine in dimethysulfoxide in an amount of 0.01 to 30% by weight, adding succinic anhydride in an amount of 1 to 5% And pyridine in an amount of 0.001 to 0.1% by volume to obtain a gemcitabine succinic acid derivative; The gemcitabine acid derivatives 0.01 to 30% by weight, di methyl amino propyl carbodiimide chilka to (N- (3-Dimethylaminopropyl) -N -ethylcarbodiimide) the covered stent such that the melt is 1 to 30 wt% per 1cm ² Immersing the coated stent in a solvent for 1 to 24 hours to obtain a covered stent coated with a gemcitabine. The method of claim 1, wherein the drug- Can be achieved by a stent manufacturing method.

Preferably, the solvent of the present invention is selected from the group consisting of water (H2O), ethanol, dimethyl acetamide (DMAc), N, N-dimethylformamide (DMF) (NMP), dimethyl sulfoxide (DMSO), tetra-hydrofuran (THF), ethylene carbonate (EC), diethyl carbonate (DEC), dimethyl carbonate dimethyl carbonate (DMC), ethyl methyl carbonate (EMC), propylene carbonate (PC), acetic acid, formic acid, chloroform, dichloromethane, Acetone, < RTI ID = 0.0 > and / or < / RTI >

Preferably, the drug coated on the covered stent of the present invention is selected from the group consisting of Paclitaxel, Docetaxel, Cisplatin, Carboplatin, Oxaliplatin, Fluorouracil (5- fluorouracil, methotrexate, doxorubicin, capecitabine, cyclophosphamide, epirubicin, mitomycin, doxycyclin, irinotecan, Etoposide, Nimustine, temozolomide, Camptothecin, Rapamycin, Etanercept, Epoeitin alfa, erythropoietin (e. G. Erythropoietin, Vincristine, Vinblastine, Cytarabine, Daunorubicin, Bleomycin; Procabazine, Tamoxifen, Leuprolide, Carmustine, Lomustine, Streptozotocin, Mitoxantrone, and Hydroxyurea. ). ≪ / RTI >

Another object of the present invention is also achieved by a drug-releasing covered stent in which an anticancer agent produced by any one of the above-mentioned methods is covalently bonded.

As described above, the present invention can have an effect of providing a stent capable of preventing post-operative restenosis and treating a local site by coating an anticancer agent for chemotherapy with a chemical method on a covered stent.

FIG. 1A is a schematic diagram showing a process of attaching cystamine to argon plasma on a covered stent according to the present invention, and FIG. 1B is a schematic diagram of covalently bonding gemcitabine to cystamine.
2 is a scanning electron micrograph of a covered stent coated with a gemcitabine prepared according to an embodiment of the present invention.
FIG. 3 is a graph showing the release rate of gemcitabine from a covered stent coated with a gemcitabine prepared according to an embodiment of the present invention. FIG. 3 (a) is an explanatory view of a device releasing the gemcitabine from the surface of the stent, FIG. 3B is a graph showing the effect of glutathione concentration on gemcitabine release, and FIG. 3C is a graph showing the effect of cell extract on gemcitabine release.
FIG. 4 is a graph showing anticancer performance against cancer cells of a covered stent coated with a gemcitabine prepared according to an embodiment of the present invention.
FIG. 5 is a graph showing anticancer activity against HuCC-T1 bile duct cancer induced rat-coated stent coated with gemcitabine prepared according to an embodiment of the present invention.

The present invention relates to a coated stent in which allyl alcohol can be bonded to a silicon surface of a covered stent using a plasma apparatus and cystamine is covalently bonded thereto to replace an amine group on the surface of the coated stent, Can be manufactured. Gemcitabine modified with succinic anhydride may be synthesized by dissolving gemcitabine dissolved in any solvent and covalently bonded to the surface of a covered stent substituted with an amine group as shown in FIG. 1B to prepare a coated stent coated with an anticancer agent.

In addition, the present invention can coat an anticancer agent on the surface of a covered stent substituted with an amine group by a physical method. For example, an anticancer agent modified with succinic anhydride can be prepared by coating the surface of a covered stent substituted with an amine group by a method of ionic bonding or the like on a covered stent.

In the present invention, usable functional chemicals on the silicon surface of the covered stent include not only allyl alcohol but also allyl amine, allyl bromide, allyl chloride, allyl alcohol, acetate, allyl glycidyl ether, allyl isothiocyanate, allyl benzyl ether, 2-Allyl-6-methylphenol ), Allyl ether, allyl methyl ether, allyl methacrylate, allyl iodide, allyl ethyl ether, allyl ether, phenyl ether, allyl chloroformate, allyl propionate, allyl 2-furoate, allyl methyl carbonate, allyl cyanide, Allyl isocyanate, Allyl disulfide, Allyl p-toluenesulfonate, Allyl butyrate, Allyl propyl ether, allyl butyl ether Allyl-proline, derivatives thereof, and the like can be used alone or in a blend. However, the present invention is not limited to these materials, and it is possible to use a substance that can bind to the surface of the stent by an allyl group no limits.

The present invention also relates to a method for preparing a coated stent to which allyl alcohol or a derivative thereof is bound and a method for producing a cysteamine, glutamine, aspartic acid, proline, lysine, , Arginine, histidine, phenylalanine, glucosamine, galactosamine, tyrosine, tryptophan, and the like may be used alone or in combination. However, The material is not limited to the above material, and there is no particular limitation as long as it is a substance that can be bound to the surface of the stent.

Examples of the solvent used in the present invention include water (H2O), ethanol, dimethyl acetamide (DMAc), N-dimethylformamide (DMF), N-methyl-2-pyrrolidinone (NMP), dimethyl sulfoxide (DMSO) (ethylene carbonate), DEC (diethyl carbonate), DMC (dimethyl carbonate), ethyl methyl carbonate (ECM), propylene carbonate, acetic acid, formic acid, chloroform, dichloromethane , DMP, and acetone.

Hereinafter, the present invention will be described with reference to examples.

A. Step of binding allyl alcohol and cystamine to the surface of the stent

The step of modifying the surface of the covered stent with an allyl alcohol, which is a chemical functional group, is performed by adjusting the vacuum of the plasma reactor to 50 mHg or less and applying the covered stent surface at a flow rate of 1 to 300 cm ³ / Lt; / RTI > for 1 minute.

The pressure in the reactor is maintained at 90 mHg, treated with allyl alcohol at a flow rate of 0.1 to 3 scm for 1 minute and 1 to 100 W glow discharge, and the surface of the covered stent is washed with distilled water. The covered aldehyde-coated stent was dried for 2 days under vacuum, and then 1,1-carbonyldiimidazole was dissolved in dimethysulfoxide at 0.1 to 10 wt%. Allyl alcohol After the combined coated stent was immersed and reacted for 3 hours, the cystamine, a linker (linker) for binding the drug after washing with diethyl ether, was dissolved in dimethylsulfoxide 0.1 to 10% by weight, and then the coated aldehyde-coated stent is immersed in this solution and reacted for 6 hours. The coated stent with cystamine is washed with diethyl ether and dried under vacuum to prepare a covered stent with exposed amine groups on its surface.

B. Synthesis of Gemcitabine Sulfate Derivatives

In order to modify gemcitabine with succinic anhydride, it is appropriate to dissolve gemcitabine in dimethysulphoxide in an amount of 0.01 to 30% by weight. If the amount exceeds 30% by weight, problems such as dissolution of gemcitabine may occur. Succinic anhydride is preferably added in an amount of 0.01 to 30% by weight to dimethysulphoxide in an amount of 30% by weight. If the amount exceeds 30% by weight, problems such as dissolution of gemcitabine may occur. Mixing an anticancer agent and succinic anhydride, adding pyridine in an amount of 0.001 to 0.1% by volume, reacting the mixture for 24 hours, and purifying the reacted mixture by silica gel chromatography.

C. Method of coating gemcitabine succinate derivatives on the surface of a covered stent

A method for coating an anticancer agent on the surface of a covered stent includes using 0.1 to 30% by weight of a gemcitabine succinic acid derivative, dimethyaminopropyl-N- (3-dimethylaminopropyl) -N-ethylcarbodiimide ) Is dissolved in an amount of 1 to 30% by weight, the coated stent is adjusted to have a weight of 0.1 to 1000 mg per 1 cm ² , and then the covered stent is immersed in a solvent for 24 hours. After completion of the reaction, the cells are washed with distilled water, lyophilized and refrigerated.

(Example 1)

Adjusting the vacuum in the plasma reactor at a 50mHg and the covered stent surface is treated with argon plasma for 5 minutes at a flow rate of 200 cm ³ / m. Allyl alcohol was injected at a flow rate of 1 sccm and treated with a 20 W glow discharge for 10 minutes.

At this time, the pressure in the reactor was maintained at 90 mHg. After the surface of the covered stent was washed with distilled water and dried for 2 days under vacuum, 1,1-carbonyldiimidazole was dissolved in dimethylsulfoxide at 2% by weight and allyl alcohol was added to the solution. The coated stent was immersed in the stent and reacted for 3 hours, followed by washing with diethyl ether.

Cystamine was dissolved in dimethylsulfoxide at 10% by weight, the covered stent activated with carbonyldiimidazole was immersed in the solution and reacted for 6 hours. As shown in FIG. 1A, cystamine To obtain a combined covered stent.

One of the anticancer drugs, gemcitabine, was dissolved in dimethysulfoxide so as to be 5 wt%, succinic anhydride was added in an amount of 5 wt%, pyridine was added in an amount of 0.001-0.1 vol% The reaction mixture was purified by silica gel chromatography to obtain gemcitabine succinic acid derivative.

5% by weight of gemcitabine succinic acid derivative and 5% by weight of N- (3-Dimethylaminopropyl) -N-ethylcarbodiimide) were dissolved in the solution. The concentration was adjusted to be 100 mg per 1 cm ² in the covered stent The coated stent was immersed in a solvent and reacted for 24 hours to obtain a covered stent coated with Gemcitabine as shown in FIG. 1B. The coated stent was observed to have a coating weight of 0.12 mg per 1 cm ² by nuclear magnetic resonance spectroscopy Respectively.

(Example 2)

FIG. 2 is a photograph of a covered stent coated with Gemcitabine prepared by the method of FIG. 1 and an electron micrograph thereof.

As shown in the picture, when the Gemcitabine was coated on the silicon surface of the covered stent, it was confirmed that it had a rough surface.

(Example 3)

The coated stent coated with gemcitabine is immersed in 10 ml of phosphate-buffered saline (pH 7.4, 0.01 M) to determine the amount of gemcitabine released from the covered stent.

As shown in FIG. 3B, when glutathione was dissolved in a phosphate buffer at a concentration of 10 mM, the release of gemcitabine was faster than when glutathione was 0 mM. In addition, when the HuCC-T1 biliary cancer cell culture fluid was mixed with phosphate buffer solution, the release rate of gemcitabine was also increased.

(Example 4)

The cytotoxicity test was carried out using the sample prepared in Example 1 above.

Biliary tract cancer cells, mainly human origin cholangiocarcinoma handle HuCC-T1 cell line (HuCC-T1) was used, 1x10 ⁴ cells, a back various concentrations of gemcitabine from 0.001 to 100 ug / ml culture in 96 well plate in HuCC-T1 cells Respectively. Cell viability was assessed by MTT assay after 24 hours incubation. Gemcitabine solution was prepared by dissolving gemcitabine in phosphate buffer and added. Gemcitabine stent was prepared by immersing the covered stent coated with gemcitabine in RPMI1640 cell culture medium to ensure sufficient release of gemcitabine. And then the covered stent was treated by immersing the covered stent without Gemcitabine in the RPMI1640 cell culture medium for the same period of time.

The covered stent itself had no effect on the growth of the cancer cells and the gemcitabine stent showed the same cancer cell growth inhibition as the gemcitabine solution.

(Example 5)

The anti-cancer activity test for solid tumor induced in rats was carried out using the sample prepared in Example 1 above. 1 x 10 ⁷ HuCC-T1 cells were subcutaneously injected into the nude mice and allowed to stand for about 2 weeks to induce solid tumors.

The covered stent coated with Gemcitabine was cut to a diameter of 1 cm, and the skin of the solid cancer part of the rat was cut and inserted under the solid tumor. After about 40 days, the cancer tissue was cut and the volume was expressed by the equation ((length * width ² ) / 2).

As shown in FIG. 5, the covered stent coated with the gemcitabine showed an excellent effect in inhibiting the growth of the cancer, compared with the covered stent without the gemcitabine coated. The ideal treatment of the digestive cancer including biliary cancer I was able to know his reign.

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed exemplary embodiments.

Claims (8)

A method of manufacturing a covered stent for a digestive arm,
Coupling allyl alcohol, which is a chemical functional group for drug binding, to the surface of the covered stent using a plasma reactor;
A cystamine-coupled covered stent, which is a chemical reactor for drug-binding, is prepared by binding cystamine to the allyl alcohol-bound covered stent using 1,1-carbonyldiimidazole. Wherein the method comprises the step of covalently bonding an anticancer agent comprising the steps of:
The method according to claim 1,
Wherein the chemical reactor for drug binding comprises:
But are not limited to, allyl alcohol propoxylate, glycidol, allylamine, butenylamine, allyl proline, allyl indole carboxylic acid, From a group consisting of allyl oxopyrrolidine carboxylic acid, allyl bromide, allyl cysteine, allyl cystamine and acryloyl cystamine. Any one or more selected,
The linker for binding the drug,
Cystamine, succinic anhydride, diethyl amine, hexamethylene diamine, adipic acid dihydrazide, succinic dihydrazide, The present invention relates to a process for the preparation of succinic dihydrazide, Oxalyldihydrazide, Carbohydrazide, Semicarbazide, Glutamic acid hydrazide, and Hydrazide Hydrazide), wherein the anticancer agent is covalently bonded to the drug-releasing coated stent.
3. The method of claim 2,
Wherein the linker is degraded by glutathione or a hydrogen ion concentration, wherein the linker is covalently bonded to the drug-releasing coated stent.
The method according to claim 1,
Wherein the plasma reactor control the vacuum of 1 to 100 mHg and processing the covered stent surface in 1-5 minutes argon plasma for a flow rate of 1 ~ 500 cm ³ / m was charged with the allyl alcohol with 0.1 ~ 10scm flow rate and 1 To 100 W of glow discharge for 1 to 60 minutes to bind the allyl alcohol to the covered stent. ≪ RTI ID = 0.0 > 18. < / RTI >
A method of manufacturing a covered stent for a digestive arm,
Dissolving gemcitabine in dimethysulfoxide in an amount of 0.01 to 30% by weight, adding 1 to 5% by weight of succinic anhydride and adding pyridine in an amount of 0.001 to 0.1% by volume to obtain gemcitabine succinic acid derivative Wow;
The gemcitabine acid derivatives 0.01 to 30% by weight, di methyl amino propyl carbodiimide chilka to (N- (3-Dimethylaminopropyl) -N -ethylcarbodiimide) the covered stent such that the melt is 1 to 30 wt% per 1cm ² Immersing the coated stent in a solvent for 1 to 24 hours to obtain a covered stent coated with a gemcitabine. The method of claim 1, wherein the drug- A method for manufacturing a stent.
6. The method of claim 5,
The solvent may be,
It can be dissolved in water (H2O), ethanol, dimethyl acetamide (DMAc), N-dimethylformamide (DMF), N-methyl-2-pyrrolidinone (NMP), dimethyl dimethyl sulfoxide (DMSO), tetra-hydrofuran (THF), ethylene carbonate (EC), diethyl carbonate (DEC), dimethyl carbonate at least one selected from the group consisting of methyl carbonate (EMC), propylene carbonate (PC), acetic acid, formic acid, chloroform, dichloromethane, Wherein the anticancer agent is covalently bonded to the drug-releasing coated stent.
6. The method of claim 5,
The drug coated on the covered stent may be,
Paclitaxel, docetaxel, cisplatin, carboplatin, oxaliplatin, 5-fluorouracil, methotrexate, doxorubicin, capecitabine, Such as capecitabine, cyclophosphamide, epirubicin, mitomycin, doxycyclin, irinotecan, etoposide, nimustine, temozolomide, but are not limited to, temozolomide, camptothecin, rapamycin, etanercept, epoeitin alfa, erythropoietin, vincristine, vinblastine, Cytarabine, Daunorubicin, Bleomycin; Procabazine, Tamoxifen, Leuprolide, Carmustine, Lomustine, Streptozotocin, Mitoxantrone, and Hydroxyurea. ), Wherein the anticancer agent is covalently bonded to the drug-releasing coated stent.
A drug-releasing coated stent covalently bound to an anticancer agent produced by one of the methods selected from any one of claims 1 to 7.

Images