KR102104068B1 - Polymer having mitochondrial targeting ability and composition for drug delivery using the polymer - Google Patents

Abstract

The present invention relates to a polymer having mitochondrial targeting ability and a composition for drug delivery using the polymer, wherein the polymer can form nanoparticles by self-assembly in a water phase even increasing a hydrophobic region of the polymer by synthesizing the polymer in which triphenylphosphonium is covalently attached to both ends of poly(ε-caprolactone) having multiple disulfide bonds or multiple diselenide bonds. In addition, the polymer can serve as a carrier which can deliver hydrophobic drugs into cells.

Description

Polymer having mitochondrial targeting ability and composition for drug delivery using the polymer {Polymer having mitochondrial targeting ability and composition for drug delivery using the polymer}

The present invention relates to a polymer having mitochondrial targeting ability at both ends of a reductively decomposable poly (epsilon-caprolactone) having multiple disulfide bonds or multiple diselenide bonds, and a composition for drug delivery using the polymer.

Biocompatible polymers are used in the development of effective systems for delivering various therapeutic agents such as chemical drugs, contrast agents, peptides, proteins, and genetic materials as constituents of nanodrug delivery systems. In particular, the position-specific nanodrug delivery system capable of targeting organs, tissues, cells, and organelles reduces the wide range of toxicity and side effects due to non-selective biodistribution, and optimizes the in vivo dynamics and distribution of the drug for therapeutic effect. It is constantly growing to maximize it. The nano-drug delivery system capable of targeting a specific location uses physiological or pathological differences such as pH, enzymes, oxidation-reduction, and chemicals at the disease site, or recognizes receptors or antigens specifically expressed on cell membranes of specific cells. Ligands or antibodies that can be used are used. However, since the actual organs of action of drugs are intracellular organelles (eg, cytoplasm, nucleus, mitochondria, Golgi apparatus, lysosomes, etc.), the intracellular organelles must be targeted to maximize the higher therapeutic efficiency.

Among the organelles, mitochondria are an intracellular power plant that metabolizes energy to convert to ATP through an oxidative phosphorylation process, and play an important role in regulating apoptosis by regulating the release of cytochrome c and other pro-apoptosis elements. do. In addition, mitochondrial dysfunction is closely related to adult diseases such as diabetes, cardiomyopathy, infertility, blindness, kidney / liver disease, stroke, etc. because it plays a role in signaling, cell differentiation, calcium concentration control, and reactive oxygen species. As mitochondrial gene mutations have been reported in various diseases such as aging, degenerative neurological diseases, and cancer diseases, research on mitochondria is increasing.

Conventionally, a method using a mitochondrial hydrophobic cation is known as a material containing a function capable of targeting mitochondria. Unlike hydrophilic cations that cannot cross the mitochondrial membrane, hydrophobic cations can pass through the mitochondrial membrane.Triphenylphosphonium (TPP) is easily accumulated on the mitochondrial substrate due to the high membrane potential difference and is chemically chemically mixed with various chemicals. Combined, it is being developed as a target drug delivery system.

Meanwhile, recently, attempts have been made to increase the efficiency of cancer treatment by encapsulating a large amount of hydrophobic drugs in the target drug delivery system as described above, and thus, research is being conducted to increase the hydrophobic region of the polymer constituting the drug delivery system. . However, it has been difficult to form nanoparticles and thus cannot be prepared as a drug delivery system.

Therefore, the target of mitochondria in cells is possible, and there is a need to study polymers capable of increasing the encapsulation efficiency of hydrophobic drugs and drug delivery systems using the same.

1. Republic of Korea Registered Patent No. 10-0950548

An object of the present invention is to provide a reducible poly (epsilon-caprolactone) polymer capable of targeting mitochondria in cells and having mitochondrial targeting ability at both ends capable of increasing the encapsulation efficiency of hydrophobic drugs.

In addition, another object of the present invention is to provide a nanoparticle using the polymer and a composition for anticancer using the same.

In addition, another object of the present invention to provide a composition for nano-drug delivery comprising the nanoparticles as an active ingredient.

In order to achieve the above object, the present invention is triphenylphosphonium (Triphenylphosphonium) is shared at both ends of a poly (epsilon-caprolactone) [poly (ε-caprolactone)] having multiple disulfide bonds or multiple diselenide bonds Provided is a reductively decomposable poly (epsilon-caprolactone) polymer having a mitochondrial targeting ability at both ends represented by Formula 1 below.

[Formula 1]

In Formula 1 A is a sulfur (S) or selenium (Se), · X is Br ^-, Cl ^-, I ^- are selected from the group consisting of, x is from 3 to 20 integer, y is a numeral from 2 to 20, .

In addition, the present invention provides nanoparticles formed by self-assembly of the polymer according to Formula 1 in a water phase.

In addition, the present invention provides an anticancer composition comprising the nanoparticles as an active ingredient.

In addition, the present invention provides a composition for nano-drug delivery comprising the nanoparticles as an active ingredient.

Reductively decomposable poly (epsilon-caprolactone) polymers having mitochondrial targeting ability at both ends according to the present invention can form nanoparticles by self-assembly in the water phase even if the hydrophobic region of the polymer is increased.

In addition, the nanoparticles prepared according to the present invention may act as an anti-cancer agent that can kill cancer cells by itself.

In addition, since a large amount of hydrophobic drugs can be enclosed inside the nanoparticles prepared according to the present invention, it can also serve as a drug delivery system that can increase drug delivery efficiency to mitochondria in cells.

1 is a view showing a synthesis process of a polymer having a reductively decomposable poly (epsilon-caprolactone); 'TMSPCL'; which has multiple disulfide bonds at which both ends have mitochondrial targeting ability.
FIG. 2 is a diagram showing ¹ H-NMR spectrum of TPP-OH Br ^- substance, MSPCL _0.5 polymer, and TMSPCL _0.5 polymer.
3 is a diagram showing the results of gel permeation chromatography (GPC) analysis of MSPCL _0.5 polymer and TMSPCL _0.5 polymer.
Figure 4 is a view of the particle size and zeta potential of the nanoparticles and DOX @ TMSPCL TMSPCL _{0.5 0.5} nanoparticles.
FIG. 5 shows apoptosis ability evaluated by MTT method after treatment of TMSPCL _0.5 nanoparticles with breast cancer cells (MCF7), which are cancer cells, and ovarian cancer cells (NCI / ADR-RES) with adreomycin resistance for 48 hours. It is the figure shown.
6 is a diagram showing the amount of doxorubicin encapsulation and the encapsulation efficiency of DOX @ TMSPCL _0.5 nanoparticles.
7 is treated with DOX and DOX @ TMSPCL _0.5 nanoparticles for 48 hours to breast cancer cells (MCF7), which are cancer cells, and ovarian cancer cells (NCI / ADR-RES) having adremycin resistance, evaluated by MTT method It is a diagram showing the cell death ability.
FIG. 8 shows the results of treating DOX and DOX @ TMSPCL _0.5 nanoparticles in breast cancer cells (MCF7), which are cancer cells, for 4 hours, and evaluating the amount of doxorubicin introduced into the cells using a flow cytometer to measure the fluorescence intensity of doxorubicin. It is a figure showing.
9 is a DOX and DOX @ TMSPCL _0.5 nanoparticles are treated with breast cancer cells (MCF7), which are cancer cells, for 4 hours, and the amount of doxorubicin introduced into the mitochondria using a flow cytometer is measured and evaluated by measuring the fluorescence intensity of doxorubicin. It is a figure showing.
FIG. 10 shows the treatment of DOX and DOX @ TMSPCL _0.5 nanoparticles with ovarian cancer cells (NCI / ADR-RES) having adrenicin resistance as cancer cells for 4 hours, followed by flow cytometry of doxorubicin introduced into the cells. It is a figure showing the result of evaluating the amount by measuring the fluorescence intensity of doxorubicin.
FIG. 11 shows the treatment of DOX and DOX @ TMSPCL _0.5 nanoparticles with ovarian cancer cells (NCI / ADR-RES) having cancer cells adrenicin resistance for 4 hours, followed by flow cytometry of the doxorubicin introduced into the mitochondria. It is a figure showing the result of evaluating the amount by measuring the fluorescence intensity of doxorubicin.
FIG. 12 shows DOX and DOX @ TMSPCL _0.5 nanoparticles treated with breast cancer cells (MCF7), which are cancer cells, for 4 hours, and the results of the distribution of doxorubicin introduced into cells, nuclei, and mitochondria using confocal microscopy are DOX, It is a diagram showing the intensity of emission fluorescence of nuclear stain reagents and mitochondrial stain reagents.
FIG. 13 shows the results of evaluating whether the DOX and DOX @ TMSPCL _0.5 nanoparticle drugs for the white arrow portion of FIG. 12 are present at the same position using a line profile for distribution in the nucleus, mitochondria, or cytoplasm. It is a drawing.
14 is a view showing the distribution of DOX and DOX @ TMSPCL _0.5 nanoparticles DOX in FIG. 12 in the nucleus, mitochondria, and other organelles using correlation coefficients.
Figure 15 is treated with DOX and DOX @ TMSPCL _0.5 nanoparticles for 4 hours to ovarian cancer cells (NCI / ADR-RES) having cancer cell adrenicin resistance, intracellular and intranuclear cells using a confocal microscope. And DOX, the nuclear staining reagent and the mitochondrial staining reagent emission intensity of the doxorubicin in the mitochondrial.
FIG. 16 shows the results of evaluating whether DOX and DOX @ TMSPCL _0.5 nanoparticle drugs for the white arrow portion of FIG. 15 are present at the same position using a line profile for distribution in the nucleus, mitochondria, or cytoplasm. It is a drawing.
17 is a diagram showing the distribution of DOX and DOX @ TMSPCL _0.5 nanoparticles DOX in FIG. 15 in the nucleus, mitochondria, and other organelles using correlation coefficients.

Hereinafter, the present invention will be described in detail.

The present inventors lengthened the region of poly (epsilon-caprolactone; PCL), a hydrophobic polymer having biocompatibility and biodegradability, introduced multiple disulfide bonds or multiple diselenide bonds, and multiple disulfide bonds. Polymers were synthesized by chemically binding TPP having a mitochondrial targeting ability to both ends of a PCL having a bond or multiple diselenide bonds.The synthesized polymer can form nanoparticles through self-assembly in the aqueous phase, It has been discovered that it can act as an anti-cancer agent that can kill cancer cells, and it can also serve as a drug delivery system that can increase the drug delivery efficiency into cells by encapsulating a large amount of hydrophobic drugs inside nanoparticles. The invention was completed.

The present invention is a poly (epsilon-caprolactone) [poly (ε-caprolactone)] triphenylphosphonium (Triphenylphosphonium) covalently bonded to both ends having a multiple disulfide bond or multiple diselenide bond Reductively decomposable poly (epsilon-caprolactone) polymers having mitochondrial targeting ability at both ends are provided.

[Formula 1]

In Formula 1 A is a sulfur (S) or selenium (Se), · X is Br ^-, Cl ^-, I ^- are selected from the group consisting of, x is from 3 to 20 integer, y is a numeral from 2 to 20, .

As an embodiment of the present invention, the polymer represented by Chemical Formula 1 is poly (epsilon-caprolactone) having multiple disulfide bonds represented by Chemical Formula 2 [multiple disulfide poly (ε-caprolactone); MSPCL] may have a covalent bond of triphenylphosphonium (TPP) to which Br ⁻ ions are bound at both ends.

[Formula 2]

In Formula 2, m is an integer from 3 to 20, and n is an integer from 2 to 20.

In addition, the present invention provides nanoparticles formed by self-assembly of the polymer in the water phase.

In addition, the nanoparticles may have an average particle diameter of 20 to 400 nm, but are not limited thereto.

As an embodiment of the present invention, a polymer (hereinafter referred to as 'TMSPCL _0.5 ') in which TPP is covalently bonded to both ends of the MSPCL represented by Chemical Formula 2 is self-assembled in the water phase to prepare TMSPCL _0.5 nanoparticles, TMSPCL _0.5 The manufacturing method of the nanoparticles is as follows, but is not limited thereto.

According to the embodiment of the present invention, after dissolving TMSPCL _0.5 polymer in DMSO, purified water was added while stirring the polymer solution. _0.5 TMSPCL polymer DMSO was removed by means of dialysis in water to meet the water is self-assembly has formed a TMSPCL _0.5 nanoparticles, into the dispersion containing the nano-particles in a dialysis membrane TMSPCL _0.5. After filtering the dispersion remaining in the dialysis membrane with filter paper, finally, TMSPCL _0.5 nanoparticles were obtained.

In addition, the present invention provides an anticancer composition comprising the nanoparticles as an active ingredient. Preferably, the cancer is breast cancer or ovarian cancer, but is not limited thereto.

In addition, the present invention provides a composition for nano-drug delivery comprising the nanoparticles as an active ingredient. Specifically, the composition can target mitochondria.

In addition, the drug is characterized in that it is a hydrophobic drug, preferably doxorubicin, SN38, paclitaxel (paclitaxel) and curcumin (curcumin) may be selected from any one or more group, more preferably doxorubicin. However, it is not limited thereto.

According to an embodiment of the present invention, TMSPCL _0.5 nanoparticles in which the TMSPCL _0.5 polymer represented by Chemical Formula 2 is self-assembled can be applied as a drug delivery system because a hydrophobic drug may be enclosed therein. In addition, the characteristic of the reduction reaction of TMSPCL _0.5 nanoparticles can be applied as a drug delivery system using the surrounding environment.

Poly (epsilon-caprolactone) [poly (ε-caprolactone)] in which a mitochondrial target triphenylphosphonium having a disulfide bond (~ S-S ~) or a diselenide bond (~ Se-Se ~) is covalently bound; PCL], the nanoparticles are stable because the concentration of glutathione is 0.02 to 2 μM outside the cell, and the concentration of glutathione in the cytoplasm and mitochondria in the cell is 1 to 11 mM, which is relatively higher than the extracellular concentration. Poly (epsilon-caprolactone) having a disulfide bond (~ SS ~) or a diselenide bond (~ Se-Se ~) in nanoparticles while being converted to a thiol group (~ SH) or selenol (~ SeH) [ poly (ε-caprolactone); PCL] is decomposed into low molecular weight PCL. The low-molecular-weight biodegradable polymer produced in this process can reduce the mitochondrial accumulation and non-specific interactions in the cell, thereby indicating biocompatibility, and the mitochondrial target reductively-decomposable nanoparticles loaded with the drug rapidly displace the drug enclosed in the cell. It can be released from the cytoplasm to improve the effectiveness of the drug.

The composition for anticancer or drug delivery according to the present invention may contain a pharmaceutically effective amount of a drug alone or may include one or more pharmaceutically acceptable carriers, excipients, or diluents. In the above, the pharmaceutically effective amount refers to an amount sufficient for the drug to be administered to animals or humans to exhibit desired physiological or pharmacological activity. However, the pharmaceutically effective amount may be appropriately changed according to the age, weight, health status, sex, administration route, and treatment period of the administration target.

In addition, "pharmacologically acceptable" in the above refers to physiologically acceptable and does not cause an allergic reaction such as gastrointestinal disorder, dizziness or similar reaction when administered to humans. Examples of the carrier, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be further included.

The composition for anticancer or drug delivery according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous or intramuscular, and the dosage of the drug is the route of administration, age, sex, weight and patient of the patient. It can be appropriately selected according to various factors such as the severity of the. In addition, the polymer composition for drug delivery of the present invention can be administered in parallel with a known compound that can increase the desired effect of the drug.

The "drug" used in the present invention is a substance capable of promoting or inhibiting a physiological function in an animal or human body to induce a desired biological or pharmacological effect, and is a chemical or biological substance suitable for administration to an animal or human. Or it means a compound, (1) prevents unwanted biological effects such as infection prevention, has a preventive effect on organic matter, (2) reduces the condition caused by disease, for example, pain or infection resulting from the disease And (3) it can play a role in alleviating, reducing or completely eliminating diseases from organic matter.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, according to the gist of the present invention, the scope of the present invention is not limited by these examples to those skilled in the art to which the present invention pertains. It will be obvious.

<Example 1> TMSPCL polymer synthesis and analysis

As shown in FIG. 1, the TMSPCL polymer includes 1 mmol of multiple disulfide poly (ε-caprolactone) carboxyl; MSPCL and 2 mmol of 2-hydroxyethyltriphenylphosphonium bromide (2-hydroxyethyltriphenylphosphoiumbromide; TPP-Br OH ^-) of the 10 ㎖ chloroform (chloroform; was prepared by dissolving the following "CHCl _3") solution (hereinafter referred to as "solution a"). Then, 4 mmol of dicyclohexylcarbodiimide (hereinafter referred to as 'DCC') and 4 mmol of 4-dimethylaminopyridine (hereinafter referred to as 'DMAP') were dissolved in 7 mL of CHCl ₃ to prepare a solution. (Hereinafter 'B solution').

After the solution B was added dropwise to the solution A, 0.1 ml of triethylamine (hereinafter referred to as 'TEA') was added, followed by stirring at room temperature for 2 days under nitrogen charge.

After completion of the reaction, dicyclohexylurea (hereinafter referred to as 'DCU'), a by-product generated using filter paper, was removed, and the solvent CHCl ₃ was removed using a rotary evaporator. The dried product was dissolved in CHCl ₃ again, precipitated in n-hexane (n-hexane), and the precipitate was vacuum dried to obtain a high molecular weight TMSPCL polymer (Formula 1, hereinafter 'TMSPCL _0.5 ').

[Formula 1]

2, the chemical structure of TPP-OH Br ⁻ , MSPCL _0.5 polymer and TMSPCL _0.5 polymer was confirmed using ¹ H-NMR, and the number of low molecular weight PCL blocks present in TMSPCL _0.5 polymer is 6.5, disulfide It was confirmed that the number of bonds was 7.5 and the number of TPPs was 2.

3, the number average molecular weights of the MSPCL _0.5 polymer and the TMSPCL _0.5 polymer were 5690 Da and 6880 Da, respectively, through gel permeation chromatography (GPC) analysis, and the polydispersity was 1.71 and 1.73, respectively. Confirmed.

<Example 2> TMSPCL _0.5  Preparation of nanoparticles, physicochemical properties and anticancer effects

To prepare TMSPCL _0.5 nanoparticles, TMSPCL _0.5 polymer was dissolved in DMSO, and purified water was added while stirring the polymer solution. TMSPCL _0.5 polymer was self-assembled with water to form TMSPCL _0.5 nanoparticles. The dispersion containing TMSPCL _0.5 nanoparticles was placed in a dialysis membrane to remove DMSO through dialysis in an aqueous phase. After filtering the dispersion remaining in the dialysis membrane with filter paper, finally, TMSPCL _0.5 nanoparticles were obtained.

The average particle size of TMSPCL _0.5 nanoparticles was 290 nm, and the average zeta potential was 29 mV due to the positive charge of TPP (FIG. 4).

The anti-cancer effect of TMSPCL _0.5 nanoparticles was spread over 5,000 breast cancer cells (hereinafter referred to as 'MCF7 cells') and ovarian cancer cells (hereinafter referred to as 'NCI / ADR-RES cells') with adreamycin resistance in a 96-well plate for 24 hours. After incubation, TMSPCL _0.5 nanoparticles were exposed for 48 hours to evaluate the cytotoxicity of cancer cells at various nanoparticle concentrations using the MTT method.

As shown in FIG. 5, TMSPCL _0.5 nanoparticles were not observed when the concentration of 50% viability in MCF7 cells and NCI / ADR-RES cells was less than 500 μg / ml.

<Example 3> DOX @ TMSPCL containing doxorubicin anticancer drug _0.5  Preparation and physicochemical properties of nanoparticles

To encapsulate the hydrophobic drug doxorubicin (hereinafter referred to as 'DOX') into TMSPCL _0.5 nanoparticles, the DOX solution corresponding to 0.5 mg is mixed with 0.1 ml of DMSO solution in which 2 mg of TMSPCL _0.5 polymer is dissolved, and this drug-polymer While stirring the solution, 4 ml of purified water was added dropwise and further stirred for 1 hour. The DOX that was not encapsulated in DMSO and nanoparticles was removed by dialysis in the water phase, and the remaining unencapsulated DOX was removed by filtration using a filter paper. The TMSPCL _0.5 nanoparticles encapsulated with the doxorubicin hydrophobic drug synthesized by the above method were designated as DOX @ TMSPCL _0.5 nanoparticles.

The prepared DOX @ TMSPCL _0.5 nanoparticles had an average particle size of 155 nm and an average zeta potential of 27.5 mV (FIG. 4).

When DOX was encapsulated inside TMSPCL _0.5 nanoparticles with the target weight content of the drug being 20% by weight, the average weight content of the actual drug of DOX @ TMSPCL _0.5 nanoparticles was 13.0% by weight, and the average encapsulation efficiency of the drug was 64.8% It was (Fig. 6).

<Example 4> DOX @ TMSPCL containing doxorubicin anticancer drug _0.5  Anti-cancer effect of nanoparticles

In order to evaluate the anti-cancer effect of DOX @ TMSPCL _0.5 nanoparticles prepared from Example 3, 5,000 MCF7 cells and NCI / ADR-RES cells were spread on a 96-well plate for 24 hours, followed by incubation of various drug concentrations in cancer cells. Having DOX @ TMSPCL _0.5 nanoparticles was treated for 48 hours to evaluate the effect of the drug by MTT method.

As shown in FIG. 7, the IC50 of DOX for MCF7 cells was about 1.52 μg / ml, and the IC50 of DOX @ TMSPCL _0.5 nanoparticles was 0.47 μg / ml. This result means that DOX @ TMSPCL _0.5 nanoparticles have 3.23 times better anticancer effect than DOX. In addition, in NCI / ADR-RES cells with adreomycin resistance, the IC50 of DOX was about 122.8 μg / ml, and the IC50 of DOX @ TMSPCL _0.5 nanoparticles was 0.70 μg / ml. This result means that DOX @ TMSPCL _0.5 nanoparticles have 175.4 times better anticancer effect than DOX.

This, NCI / ADR-RES cells have adremycin resistance, and DOX flows into the cell by diffusion. At this time, the IC50 value of the drug is increased by being discharged out of the cell by an efflux pump, but DOX It is confirmed that @TMSPCL _0.5 nanoparticles have a much better anti-cancer effect because they are not affected by the discharge pump because they are introduced into the cell through the pathway through the endosome.

<Example 5> DOX @ TMSPCL _0.5  Drug influx and drug distribution into cells and organelles of nanoparticles

MCF7 cells and NCI / ADR-RES cells (5 ×) to evaluate DOX @ TMSPCL _0.5 nanoparticles prepared from Example 3 above for intracellular drug intake (Cellular uptake; CU) and mitochondrial uptake (MU) 10 ⁵ cells / 2 ml) on a 6-well plate and incubated for 24 hours, then treated with DOX and DOX @ TMSPCL _0.5 nanoparticles ([DOX] = 1 μg / ml) for 4 hours and their cells and organelles Each drug inflow was analyzed.

To evaluate the intracellular influx (CU) of DOX and DOX @ TMSPCL _0.5 nanoparticles, cells were washed twice with DPBS and separated with trypsin-EDTA (trypsin-EDTA). Mitochondrial influx (MU) is separated according to the protocol of the Mitochondrial Fractionation Kit, and the FACSCanto ^TM Ⅱ flow cytometer (Flow cytometer, Becton-Dickinson, Franklin Lakes, Nj, USA) and primary argon laser (primary) Argon laser) was used to measure and analyze the fluorescence of drugs introduced into the cells.

As a result, when the intracellular drug influx (CU) of DOX was set to 1 in MCF7 cells as shown in FIG. 8, the intracellular drug influx (CU) of DOX @ TMSPCL _0.5 nanoparticles was 2.25 ± 0.12. In addition, as shown in FIG. 9, when the drug influx (MU) in the mitochondria of DOX was set to 1, the drug influx (MU) in the mitochondria of DOX @ TMSPCL _0.5 nanoparticles was 3.49 ± 0.33.

In addition, as shown in FIG. 10, when the intracellular drug influx (CU) of DOX was set to 1 in NCI / ADR-RES cells, the intracellular drug influx (CU) of DOX @ TMSPCL _0.5 nanoparticles was 13.14 ± 2.18. , When the drug influx (MU) in the mitochondria of DOX was set to 1, the drug influx (MU) in the mitochondria of DOX @ TMSPCL _0.5 nanoparticles was 34.15 ± 2.88 (FIG. 11).

In addition, MCF7 cells and NCI / ADR-RES cells were placed in a confocal dish and incubated for 24 hours to confirm the distribution of the organelles of the delivered DOX according to the use of the DOX @ TMSPCL _0.5 nanoparticle drug delivery system. , DOX and DOX @ TMSPCL _0.5 nanoparticles and further incubated for 4 hours. Before the end of the experiment, Hoechst 33342 (5 μg / ml) or SYTO ⁴⁰ (500 nM) was treated for 10 minutes to stain the nucleus, and MitoTracker Deep Red FM (200 nM) was treated for 15 minutes to stain the mitochondria. Washed twice with DPBS, and the distribution of organelles and fluorescence of doxorubicin are excitation lasers (405 nm diode, 543 nm HeNe, 633 nm HeNe) and bandpass emission filters (LSM710; Carl) Zeiss, Oberkochen, Germany).

FIG. 12 is a fluorescence image of DOX and DOX @ TMSPCL _0.5 nanoparticles introduced into the nucleus and mitochondria of MCF7 cells. FIG. 13 is data showing the white arrows in FIG. 12 as fluorescence intensities of DOX and DOX @ TMSPCL _0.5 nanoparticles introduced into the nucleus and mitochondria of MCF7 cells. 14 shows the distribution of nuclei, mitochondria, and other organelles in FIG. 12 using correlation coefficients. Through these results, it was confirmed that DOX as well as DOX delivered by DOX @ TMSPCL _0.5 nanoparticles are mainly distributed in the mitochondria rather than the nucleus, and DOX @ TMSPCL _0.5 nanoparticles have better drug inflow efficiency into the mitochondria than DOX.

FIG. 15 is a fluorescence image of DOX @ TMSPCL _0.5 nanoparticles introduced into the nucleus and mitochondria of NCI / ADR-RES cells. FIG. 16 is data showing the white arrow of FIG. 15 as fluorescence intensity of DOX @ TMSPCL _0.5 nanoparticles introduced into the nucleus and mitochondria of NCI / ADR-RES cells. 17 shows the distribution of nuclei, mitochondria, and other organelles of FIG. 15 using correlation coefficients. Through these results, DOX was all leaked by the effluent pump, so the distribution of DOX drug was hardly seen. DOX delivered by DOX @ TMSPCL _0.5 nanoparticles is mainly distributed in the mitochondria rather than the nucleus. It was confirmed that this is superior.

Accordingly, it was confirmed that the DOX @ TMSPCL _0.5 nanoparticles prepared according to the present invention are capable of targeting mitochondria in both sensitive and water-resistant cells, and have high anticancer effects due to excellent drug inflow efficiency.

Claims (9)

An amount represented by the following formula 1, in which triphenylphosphonium is covalently attached to both ends of poly (epsilon-caprolactone) [poly (ε-caprolactone)] having multiple disulfide bonds or multiple diselenide bonds Reductively decomposable poly (epsilon-caprolactone) polymer with mitochondrial targeting ability at the terminal:
[Formula 1]

In Formula 1 A is a sulfur (S) or selenium (Se), · X is Br ^-, Cl ^-, I ^- are selected from the group consisting of, x is from 3 to 20 integer, y is a numeral from 2 to 20, . Nanoparticles formed by self-assembly of the polymer according to claim 1 in the water phase. According to claim 2,
The nanoparticles,
Nanoparticles, characterized in that the average particle diameter is 20 to 400nm. An anti-cancer composition comprising the nanoparticle according to claim 2 as an active ingredient. The method of claim 4,
The cancer is anti-cancer composition, characterized in that the breast or ovarian cancer. A composition for nano-drug delivery, comprising the nanoparticle according to claim 2 as an active ingredient. The method of claim 6,
The drug is a nano drug delivery composition, characterized in that the hydrophobic drug. The method of claim 7,
The hydrophobic drug,
Doxorubicin, SN38, paclitaxel, and nano-drug delivery composition characterized in that any one or more selected from the group consisting of curcumin. The method according to any one of claims 6 to 8,
The composition is a composition for nano-drug delivery, characterized in that it targets mitochondria.

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