KR20210118581A - Amphiphilic polymers, drug delivery system and tumor treatment system using the same - Google Patents

Abstract

The present invention relates to an amphiphilic polymer compound, and a drug carrier and a tumor treatment system using the same. More specifically, the amphiphilic polymer compound of the present invention contains a thioketal linker, thereby being able to be decomposed selectively and efficiently by sensitively reacting to reactive oxygen species (ROS), and forming polymer micelles in an aqueous phase, it is possible to provide a drug carrier that contains a drug in a hydrophobic core, has excellent selective accumulation into target cancer cells, has low toxicity, and is capable of photodynamic therapy and chemical drug therapy at the same time. Another objective of the present invention is to provide the tumor treatment system using the same.

Description

Amphiphilic polymers, drug delivery system and tumor treatment system using the same {Amphiphilic polymers, drug delivery system and tumor treatment system using the same}

The present invention relates to an amphiphilic polymer compound, a polymer micelle using the same, a drug delivery system using the same, and a tumor treatment system using the same.

One of the recent promising approaches for anticancer treatment is chemo-photodynaic therapy, which combines chemotherapy and photodynamic therapy (PDT).

Chemotherapy is a method of treating cancer using drugs such as anticancer drugs, and has many problems in determining an appropriate concentration for the death of cancer cells and survival of normal cells. In order to more effectively treat cancer cells, administration of high concentrations of anticancer agents can induce apoptosis even in normal cells.

In addition, photodynamic therapy (PDT) is a highly reactive oxygen species, singlet oxygen ( ¹ O ₂ ) by irradiating light to a photosensitizer (PS) that has selectivity and photosensitivity for various tumors. It is a method to selectively attack and destroy only cancer cells or various tumor tissues by generating reactive oxygen species (ROS), including These photosensitizers have a property of specifically accumulating in cancer tissues, and when irradiated with light of a specific wavelength after a certain time has elapsed, only cancer tissues can be necrotic and normal tissues can be preserved. However, photodynamic therapy has problems in that light transmittance is limited, phototoxicity, and concentration of the photosensitizer in the tumor must be controlled.

In the chemo-photodynamic therapy method, it is required to develop a drug delivery system that can effectively apply and maximize the advantages of the grafting of chemotherapy and photodynamic therapy while reducing the disadvantages of each of these chemotherapy and photodynamic therapy. have. In addition, there is a demand for the development of a new treatment method capable of obtaining an excellent selective treatment effect on cancer cells while remarkably reducing side effects by increasing selectivity for cancer cells by using the same.

Republic of Korea Patent Publication No. 10-2018-0081494 (2018.07.16)

It is an object of the present invention to provide an amphiphilic polymer compound, a drug delivery system and a drug delivery system comprising a polymer micellar using the same.

Another object of the present invention is to provide an amphiphilic polymer compound having excellent sensitivity to reactive oxygen species (ROS).

Another object of the present invention is to include a hydrophobic drug and a photodynamic compound in the hydrophobic core of a polymer micellar formed by self-assembly of the amphiphilic polymer compound, have excellent cell penetration, and have long-term circulation in the blood and selective in tumor cells. An object of the present invention is to provide a drug delivery system having an excellent accumulation rate.

Another object of the present invention is that the polymer micelle primarily reacts with active oxygen species to release some hydrophobic drugs and photodynamic compounds, and secondarily reacts with active oxygen species generated by light irradiation to form hydrophobic drugs and photodynamics It is to provide a drug delivery system capable of simultaneously effectively expressing the characteristics of chemotherapy and photodynamic therapy by releasing all of the compounds to cause local high concentrations of hydrophobic drugs and reactive oxygen species to target tumor cells.

Another object of the present invention is to provide a tumor treatment system comprising the drug delivery system and a near-infrared irradiator.

The present invention for achieving the above object is characterized in that one end of the hydrophilic water-soluble polymer and one end of the compound having a hydrophobic hydrocarbon group are covalently bonded with a thioketal linker (TL), an amphiphilic polymer compound.

In one embodiment of the present invention, the hydrophilic water-soluble polymer is polyethylene glycol, acrylic acid-based polymer, methacrylic acid-based polymer, polyamidoamine, poly(2-hydroxypropyl methacrylamide)-based polymer, water-soluble polypeptide, water-soluble polysaccharide or polyglycerol etc. may be.

In one aspect of the present invention, the hydrocarbon group of the compound having a hydrophobic hydrocarbon group may be an aliphatic hydrocarbon group of (C12 to C22).

In one aspect of the present invention, the ratio of the weight average molecular weight of the hydrophilic water-soluble polymer to the molecular weight of the compound having a hydrophobic hydrocarbon group may satisfy the following formula (1).

[Equation 1]

(In Formula 1, WSP means the weight average molecular weight of the hydrophilic water-soluble polymer, and HC means the molecular weight of the compound having a hydrophobic hydrocarbon group)

In one embodiment of the present invention, one terminal of the hydrophilic water-soluble polymer and one terminal of the compound having a hydrophobic hydrocarbon group may be covalently bonded to a thioketal linker with an amide group, respectively.

In one aspect of the present invention, the amphiphilic polymer compound may be polyethylene glycol-TL-(C12~C22)alkylamine.

Another aspect of the present invention is characterized in that the amphiphilic polymer compound is a polymer micelle formed by self-assembly in an aqueous phase.

In another aspect of the present invention, the polymer micelle is a drug delivery system comprising a hydrophobic drug and a photodynamic compound in a hydrophobic core.

In one embodiment of the present invention, the hydrophobic drug is a hydrophobic anticancer compound, and the thioketal linker may be reactive by reactive oxygen species.

In one aspect of the present invention, the photodynamic compound is any one selected from the group consisting of porphyrin-based compounds, chlorine-based compounds, bacteriochlorine-based compounds, phthalocyanine-based compounds, naphthalocyanine-based compounds, and 5-aminolevulin ester-based compounds. It may include one or two or more.

As an aspect of the present invention, the drug delivery system primarily releases a portion of the hydrophobic drug in response to reactive oxygen species around the cell membrane, and disrupts the polymer micelle in response to the reactive oxygen species generated by light irradiation to cause a hydrophobic core It may be characterized in that all of the hydrophobic drug contained in the secondary release.

Another aspect of the present invention is characterized in that the tumor treatment system comprising a near-infrared irradiator and the drug delivery system.

As an aspect of the present invention, the light irradiated from the near-infrared irradiator may be absorbed by the photodynamic compound included in the drug delivery system to generate reactive oxygen species.

The amphiphilic polymer compound according to the present invention contains a thioketal linker, and thus reacts sensitively to reactive oxygen species, and can be selectively and efficiently decomposed. In addition, by forming a polymer micelle in the aqueous phase and including the drug and the photodynamic compound in the hydrophobic core, the selective accumulation into the target cancer cell is excellent, and a drug that can prevent side effects such as toxicity caused by accumulating in the body for a long time A carrier may be provided. The drug delivery system primarily reacts rapidly to reactive oxygen species, a part is decomposed, and secondarily, the remaining drug delivery system is immediately decomposed by near-infrared irradiation, and a large amount of drug and photodynamic compound are released to target cells. , is provided as a therapeutic agent for chemo-photodynamic therapy capable of maximizing the cancer treatment effect by lowering the apoptosis or activity of cancer cells, and may also provide a tumor treatment system using the same.

1 is a schematic diagram showing the mechanism of action of polyethylene-TL-stearamine (PTS) according to Example 1 of the present invention and the drug delivery system according to Examples 2 to 4;
2 shows a reaction scheme for preparing the amphiphilic polymer compound of the present invention.
^{3 shows 1} H-NMR spectrum and FT-IR spectrum, respectively, ^{measured using 1} H-NMR and FT-IR spectroscopy of the PTS prepared in Example 1 of the present invention.
4 shows the average hydrodynamic diameter and zeta potential of micelles prepared in Examples 1 to 8 of the present invention, and FE-TEM images of micelles prepared in Example 6, respectively.
5 shows the emission characteristics of DOX and PhA of PTS-DP prepared in Example 6 of the present invention, respectively.
Figure 6 shows the anticancer properties of PTS-DP prepared in Example 6 of the present invention under in vitro conditions, Figure 6a shows the viability of CT26 cells, Figure 6b shows the intracellular ROS, Figure 6c and Figure 6d shows the measurement of the production rate of caspase 3.
7 shows the biocompatibility of the PTS-DP prepared in Example 6 of the present invention, FIG. 7a shows the in vivo distribution of PTS-example, and FIG. 7b shows the in vitro fluorescence imaging of the tumor. FIG. 7c shows the change in concentration with time after PTS-DP injection, and FIG. 7d shows the concentration of DOX by light irradiation of PTS-DP.
Figure 8 shows the tumor treatment effect of the PTS-DP prepared in Example 6 of the present invention, Figure 8a shows the change in tumor volume over time, Figure 8b shows the average tumor weight change.
9 is a fluorescence microscopy image showing the generation of ^{singlet oxygen ( 1} O ₂ ) before and after light irradiation of PTS-DP prepared in Example 6 of the present invention.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. The drawings introduced below are provided as examples in order to sufficiently convey the spirit of the present invention to those skilled in the art. Therefore, the present invention is not limited to the drawings presented below and may be embodied in other forms, and the drawings presented below may be exaggerated to clarify the spirit of the present invention.

Also, unless defined otherwise, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description herein is for the purpose of effectively describing particular embodiments only and is not intended to limit the invention.

Also, the singular forms used in the specification and appended claims may also be intended to include the plural forms unless the context specifically dictates otherwise.

In addition, when a part "includes" a certain component, this means that other components may be further included rather than excluding other components unless otherwise stated.

The term 'photodynamic therapy' used in the present invention has the same meaning as 'photodynamic therapy'.

The term 'active oxygen species' used in the present invention refers to highly reactive oxygen species such as 'reactive oxygen species' and 'ROS', and includes 'singlet oxygen', 'singlet oxygen', and ' ¹ O ₂ '. concept that includes

The term 'photodynamic compound' used in the present invention means 'photosensitizer'.

As used herein, the term 'cancer cell' has the same meaning as 'tumor', 'tumor cell', and 'cancer'.

The term 'drug' used in the present invention is a concept including 'hydrophobic drug'.

The term 'collapse' used in the present invention is a concept including the contents of 'decomposition' of the binding portion of the compound.

The present invention for achieving the above object provides an amphiphilic polymer compound, a drug delivery system and a drug delivery system comprising a polymer micellar using the same.

The present inventors have deepened research on drug delivery systems that have a high accumulation rate for target cells, minimize toxicity to normal cells, and can effectively implement chemo-photodynaic therapy, and hydrophilic water-soluble polymers By synthesizing an amphiphilic polymer compound formed by covalently bonding a hydrophobic hydrocarbon group with a thioketal group, it was confirmed that the above effects can be realized, and the present invention has been completed.

Hereinafter, the amphiphilic polymer compound according to an aspect of the present invention will be described in detail.

In one aspect of the present invention, the amphiphilic polymer compound may be one in which one end of the hydrophilic water-soluble polymer and one end of the compound having a hydrophobic hydrocarbon group are covalently bonded with a thioketal linker (TL).

The hydrophilic water-soluble polymer means a polymer having hydrophilicity and high solubility in water, and the weight average molecular weight may be in the range of 500 to 10,000 g/mol, preferably 800 to 5,000 g/mol, and more Preferably, it may be 1,000 to 3,000 g/mol. The hydrophilic water-soluble polymer is not limited, but for example, polyethylene glycol, acrylic acid-based polymer, methacrylic acid-based polymer, polyamidoamine, poly(2-hydroxypropyl methacrylamide)-based polymer, water-soluble polypeptide, water-soluble polysaccharide, poly It may include glycerol or derivatives thereof.

The water-soluble polypeptide is not particularly limited, but may be one selected from polyglutamic acid, polyaspartic acid, polyaspartamide, and copolymers thereof, but is not limited thereto. .

The water-soluble polysaccharide is specifically hyaluronic acid, alginic acid, dextran, pectin, chondroitin sulfate, heparin, hydroxypropyl cellulose, hydroxyethylcellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose acetate succinate and its It may be selected from derivatives, but is not limited thereto.

The compound having the hydrophobic hydrocarbon group is a compound containing a hydrocarbon group having hydrophobic properties, and the hydrocarbon group may have an aliphatic hydrocarbon group of (C12-C22), specifically, having a straight-chain aliphatic hydrocarbon group of (C12-C22). can The hydrocarbon group may specifically include a hydrocarbon group such as a (C12 to C22) alkyl group, (C12 to C22) alkenyl group, or (C12 to C22) alkynyl group.

The compound having the hydrophobic hydrocarbon group is specifically hexadecylamine, heptadecylamine, stearamine, nonadecylamine, dodecylamine, bis(2-hydroxyethyl). ) Laurylamine (N,Nbis(2-hydroxyethyl)laurylamine), linoleic acid, linolenic acid, palmitic acid, oleylamine, hexadecanoic acid , stearic acid, oleic acid, eicosencic acid, and erucic acid may be selected from.

The compound having the hydrophobic hydrocarbon group may have a weight average molecular weight of 100 to 500 g/mol, preferably 150 to 350 g/mol, and more preferably 180 to 300 g/mol. However, it is not limited thereto.

The linking group connecting the hydrophilic water-soluble polymer and the compound having a hydrophobic hydrocarbon group may be covalently bonded using a thioketal linker (TL), and specifically, having one terminal of the hydrophilic water-soluble polymer and a hydrophobic hydrocarbon group Each end of the compound may be covalently bonded to a thioketal linker with an amide group.

The thioketal linker has very high sensitivity to reactive oxygen species (ROS) and excellent binding stability under normal physiological conditions, so that the drug carrier described later can prevent premature drug leakage during circulation in the blood. It is more preferable to have In addition, the polymer micelles, which will be described later, have excellent reactivity with respect to active oxygen species formed by near-infrared irradiation, so that drug release can be accelerated, which is preferable.

Preferably, a specific aspect of the amphiphilic polymer compound may be polyethylene glycol-TL-(C12~C22)alkylamine.

A ratio of the weight average molecular weight of the hydrophilic water-soluble polymer to the compound having a hydrophobic hydrocarbon group may satisfy the following formula (1).

[Equation 1]

(In Formula 1, WSP means the weight average molecular weight of the hydrophilic water-soluble polymer, and HC means the molecular weight of the compound having a hydrophobic hydrocarbon group)

의 값은 0.01 내지 0.3, 구체적으로는 0.04 내지 0.25, 더욱 구체적으로는 0.08 내지 0.2인 것일 수 있다. 상기 식 1을 만족하는 친수성 수용성 고분자와 소수성 탄화수소기를 가지는 화합물로 형성된 양친성 고분자는, 종래의 양친성 고분자와 달리, 소수성 탄화수소기의 분자량이 작아, 소수성 약물과 광역학 화합물을 포함하는 수상에서 자기조립성이 우수하며, 제조되는 고분자 미쉘이 속도론적으로 제어(kinetically controlled)되기보다 열역학적으로 제어(thermodynamically controlled)됨으로써, 재현성이 우수한 장점을 가진다. 더욱이, 생성되는 고분자 미쉘의 소수성 영역의 분자량이 작아, 2차 입자로 응집되지 않으며 분산성 및 안정성이 더욱 우수하고, 입자 크기가 작으며, 현저히 향상된 콜로이드 안정성을 가질 수 있어 바람직하다.In Equation 1 above

The value of may be 0.01 to 0.3, specifically 0.04 to 0.25, and more specifically 0.08 to 0.2. The amphiphilic polymer formed of a hydrophilic water-soluble polymer satisfying Formula 1 and a compound having a hydrophobic hydrocarbon group, unlike the conventional amphiphilic polymer, has a small molecular weight of the hydrophobic hydrocarbon group, Assembleability is excellent, and the polymer micelles produced are thermodynamically controlled rather than kinetically controlled, and thus have excellent reproducibility. Moreover, the molecular weight of the hydrophobic region of the resulting polymer micelles is small, it is not agglomerated into secondary particles, the dispersibility and stability are better, the particle size is small, and it is preferable because it can have significantly improved colloidal stability.

Hereinafter, a polymer micelle, a drug delivery system, and a tumor treatment system using the same according to an embodiment of the present invention will be described in detail.

The drug delivery system according to an aspect of the present invention has the feature that photodynamic treatment and chemical drug treatment are possible at the same time.

In one aspect of the present invention, the polymer micelle is an amphiphilic compound in which one end of the hydrophilic water-soluble polymer and one end of the compound having a hydrophobic hydrocarbon group are covalently bonded with a thioketal linker (TL) in an aqueous phase, as described above. It may be formed by self-assembly.

In a preferred embodiment, when the compound having a hydrophobic hydrocarbon group bonded by the thioketal linker (TL) is a compound having an aliphatic hydrocarbon group of (C12 to C22), the amphiphilic polymer compound is a thermodynamically uniform and stable polymer micelle. can be formed easily. The polymer micelles include a hydrophilic shell part and a hydrophobic core part, and the aliphatic hydrocarbon group of (C12 to C22) forms a hydrophobic core part. The hydrophobic core part can stably capture hydrophobic drugs and photodynamic compounds, and has excellent sensitivity to the environment of the aqueous phase in which the polymer micelle is located, so it is preferable to implement functionality as a drug delivery system with excellent target selectivity and release properties .

Specifically, as the aliphatic hydrocarbon group of (C12 to C22) forms the hydrophobic core, the hydrophobic core may have a certain degree of fluidity. This fluidity has an advantageous advantage compared to the hydrophobic core part having low fluidity formed by the hydrophobic polymer in that it can rapidly release the hydrophobic drug located in the hydrophobic core in response to the active oxygen species. According to a preferred aspect of the present invention, the long-chain aliphatic hydrocarbon group forms a hydrophobic core, thereby providing sufficient hydrophobicity to the hydrophobic core to stably trap hydrophobic drugs and provide a certain degree of fluidity, thereby degrading active oxygen species (ROS- labile) can be easily degraded by reacting sensitively to reactive oxygen species around the cell membrane.

In addition, compared to the case of using a hydrophobic polymer having a large molecular weight, even if the size of the micelles formed by self-assembly in the water phase has a relatively small size, a certain degree of fluidity provided by the (C12~C22) aliphatic hydrocarbon group. Therefore, it is preferable in that a sufficient amount of hydrophobic drug can be stably captured in the hydrophobic core.

The polymer micelle according to the present invention may include active ingredients such as a hydrophobic drug and a photodynamic compound in a core portion having hydrophobicity, and may be preferably used as a drug carrier.

The hydrophobic drug is intended to interfere with macromolecular biosynthesis in target cells, and is not limited as long as it is used as an anticancer compound, but specifically paclitaxel, doxorubicin, gemcitabine, cilolimus, etoposide, vinblastine, vinca alkaloid , docetaxel, cisplatin, gleevec, adriamycin, cyclophosphamide, teniposide, 5-fluorouracil, camptocecin, tamoxifen, anasterozole, floxuridine, leuprolide, flotamide, zoledronate , vincristine, streptozotocin, carboplatin, topotecan, belotecan, irinotecan, vinorelbine, hydoxyurea, valrubicin, retinoic acid series, mesotrexate, mechlorethamine, chlorambucil , busulfan, doxyfluridine, prednisone, testosterone, mitoxantrone, aspirin, salicylate, ibuprofen, naprosen, fenoprofen, indomethacin, phenylbutazone, cyclophosphamide, mecloethamine , dexamethasone, celecoxib, valdecoxib, nimesulide, cortisone, corticosteroids and mitomycin C may be one comprising any one or two or more selected from the group consisting of.

The photodynamic compound generates singlet oxygen upon irradiation with light, so that the singlet oxygen interacts with neighboring cell membranes to kill cells, and is generally a material used as a photosensitizer (PS). Although not limited, specifically, any one or two or more selected from the group consisting of a porphyrin-based compound, a chlorine-based compound, a bacteriochlorine-based compound, a phthalocyanine-based compound, a naphthalocyanine-based compound, and a 5-aminolevulin ester-based compound may be doing

The drug carrier may include 1 to 30 parts by weight of the mixture of the hydrophobic drug and the photodynamic compound with respect to 100 parts by weight of the polymer micelle, preferably 5 to 25 parts by weight, more preferably 10 to 25 parts by weight. It may be included in 20 parts by weight. In addition, the hydrophobic drug and the photodynamic compound included in the hydrophobic core portion of the polymer micelle may be included in a weight ratio of 1: 2 to 2: 1. In the above range, the hydrodynamic diameter measured using dynamic light scattering (DLS) analysis of polymer micelles has a small size of 50 to 200 nm, specifically 80 to 150 nm, and it is preferable because it can have better colloidal stability. .

The light irradiated to the photodynamic compound is not limited as long as it is light in a wavelength band capable of activating the photodynamic compound, but may preferably be near intra rea (NIR). The near-infrared light has a deep penetrating power to the tissue, and is preferable because it has low scattering and very low phototoxicity.

According to a preferred aspect of the drug delivery system according to the present invention, a long-chain aliphatic hydrocarbon group is bonded to a hydrophilic water-soluble polymer through a thioketal linker having a degradability (ROS-labile) to an active oxygen species, and as described above, a stable polymer micellar to form The thioketal linker may be easily decomposed in response to reactive oxygen species in the surrounding environment where the polymer micelles are located. As the hydrophilic water-soluble polymer and the compound containing a long-chain aliphatic hydrocarbon group are separated by the decomposition reaction, each molecule loses its amphiphilicity, and the colloidal stability of the polymer micelle may be rapidly reduced. The polymer micelles with poor colloidal stability cannot stably capture the hydrophobic drug, so that the hydrophobic drug is primarily released.

On the other hand, when near-infrared rays are irradiated to the tissue site where the drug carrier is located, the photodynamic compound can absorb the near-infrared rays and release reactive oxygen species in a large amount in the hydrophobic core. In polymer micelles with poor colloidal stability or stably existing polymer micelles due to the surrounding oxidizing environment, most of the thioketal linkers are decomposed by active oxygen species generated in the hydrophobic core, so that most of the polymer micelles are substantially collapsed and hydrophobic The hydrophobic drug located in the core may be released secondarily.

The large-scale release effect of a hydrophobic drug by near-infrared irradiation may be more remarkable because a long-chain aliphatic hydrocarbon group forms a hydrophobic core. This effect may be derived from the fact that the long-chain aliphatic hydrocarbon group has a certain degree of fluidity and has micellar properties rather than particle properties. On the other hand, when the hydrophobic core part is formed of a hydrophobic polymer, the fluidity of the hydrophobic core part is low, and it has particle properties rather than micellar properties, so even if the thioketal linker is decomposed, the hydrophobic polymer still exists as particles, so that the rapid release of the hydrophobic drug is reduced. It is not preferable because it can be inhibited. That is, the drug delivery system is formed of an amphiphilic compound having a hydrophobic hydrocarbon group having a carbon number (C12 to C22) that satisfies Formula 1 above, and thus, unlike the case of using a polymer, the size of the polymer micelles is small and does not aggregate, Monodispersibility and colloidal stability are further improved, which is preferable. Moreover, when light is irradiated to the lesion, the polymer micelles remaining without being destroyed by the additionally generated reactive oxygen species (ROS) are immediately decomposed, and the hydrophobic drug and photodynamic compound inside are released in a large amount within a short time. By being locally present in a high concentration in the target cancer cells, it is more preferable to effectively reduce or kill the activity of cancer cells, and it is applicable as an effective therapeutic agent for tumors.

The drug delivery system according to an embodiment of the present invention having the above characteristics is provided with a near-infrared irradiator and can be used as a preferred tumor treatment system.

Hereinafter, the present invention will be described in detail by way of Examples. The examples described below are only for helping the understanding of the present invention, and the present invention is not limited thereto.

Hereinafter, the present invention will be described in more detail based on Examples and Comparative Examples. However, the following Examples and Comparative Examples are merely examples for explaining the present invention in more detail, and the present invention is not limited by the following Examples and Comparative Examples.

[Test method]

1. Determination of thioketallinker (TL) degradation

The micelles prepared in the following Examples were suspended in a phosphate-buffered saline (PBS) having a pH of 7.4 and a PhA concentration of 3 μg/ml, and then irradiated with an intensity of ^{100 mW cm -2 for different time intervals.} .

Free thiol groups were determined by 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) analysis.

The fluorescence intensity of doxorubicin (DOX) emitted under light irradiation at different time intervals was measured using a SPARK 10M multimode microplate reader (Tecan, Austria) according to the recorded fluorescence spectra at an excitation wavelength of 470 nm and an emission wavelength of 500 to 700 nm. It was decided.

2. Measurement of singlet oxygen generation (SOG)

It was measured using dimethylanthracene (DMA), which irreversibly reacts with ¹ O ₂ in an organic solution and water.

20 mM DMA was mixed with PTS-DP prepared in Examples below in DMF and phosphate-buffered saline (PBS), and 670 nm light was irradiated with 100 mW cm ^-2 intensity at different time intervals. The fluorescence intensity of DMA was measured using a SPARK 10M microplate reader (Tecan, Austria) at an excitation wavelength of 360 nm and an emission wavelength ranging from 380 to 550 nm.

3. Determination of in vitro release of DOX

In order to evaluate the release of DOX from the polymer micelles upon light irradiation, PTS-DP prepared in the following example was dispersed in PBS and irradiated with light of a wavelength of 670 nm at 100 mW cm ^-2 intensity for 30 minutes. All samples were placed in a dialysis bag, sealed, and stored in 50 mL of release medium (1 x PBS) under dark conditions while stirring at 200 rpm. To mimic the intracellular ROS environment, an appropriate amount of H ₂ O ₂ was added to release the medium to adjust the final H ₂ O ₂ concentration to 100 μM. 1 ml of release medium was collected at different time intervals and replaced with an equal volume of fresh medium. The concentration of DOX in the release medium was then quantified by high performance liquid chromatography (HPLC) analysis. A dichloromethane (DCM)-based extraction method was used for the extraction of hydrophobic PhA. Briefly, an equal volume of DCM was added to 1 ml of released media collected at each time interval and the resulting solution was thoroughly mixed. After collecting the DCM phase containing PhA and drying the DCM, the concentration of PhA was determined by HPLC analysis.

4. Cell viability and combination index (CI) analysis

The in vitro cell viability profile of PTS-DP prepared in the Examples below was evaluated using a mouse colon cancer cell line (CT-26). Briefly, 1×10 ⁴ CT-26 cells were seeded in 96-well plates at a density of ^{1 ×10 5} cells mL _{−1 and cultured at 37° C. and 5% carbon dioxide (CO 2} ) atmosphere for 16 hours. The culture medium was then aspirated, samples were added at various concentrations, and the plates were further incubated at 37° C. for 24 hours. Cell viability profiles were then quantified using the CCK-8 assay according to the manufacturing protocol.

After chemo-photodynamic treatment treatment, the level of expression of Caspase 3, an apoptosis marker, was evaluated using Western blot analysis. Briefly, cells were suspended overnight in lysis buffer at -80 °C for 24 h. The supernatant was collected and the protein concentration was measured using a bicinchoninic acid assay kit (Thermo Scientific, Waltham, MA, USA). Protein extracts were separated using 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was blocked using 5% skim milk and treated with primary antibody (Anti Caspase-3 Antibody, 1:1000, Cell Signaling Technology, MA, USA, Danvers, MA and anti-β antibody (1:5000, Santa Cruz Biotechnology, primary antibody) After the membrane was further incubated with a horseradish peroxidase-conjugated antibody, blots were visualized using SmartChemi (Korea Lab Tech, Korea).

IC50 levels were calculated graphically and mean IC50 levels were derived. Results were analyzed using the Calcusyn software program. The results of sequential treatment by DOX and PhA were analyzed according to the method of Chou and Talylay using the Calcusyn software program (Biosoft, Cambridge, UK). The resulting combination index (CI) is a quantitative measure of the degree of interaction between different drugs. (additive when CI is equal to 1; antagonism when CI is greater than 1; synergistic when C1 is 1 to 0.7; synergistic when C1 is 0.7 to 0.3; CI value less than 0.3, strong It indicates synergy.)

5. Cell Uptake and Live Cell Imaging Measurements

To investigate the cellular uptake of PTS-DP prepared in Examples below, 1 × 10 ⁴ CT-26 cells were cultured in a Lab-Tek chamber chamber and cultured at 37° C. and 5% CO ₂ conditions for 16 hours. Then the medium was removed and DOX (5 μg ml ^-1 ), PhA (8 μg ml ^-1 ) or PTS-DP (10 μg ml ^-1 ) in DMEM was added and further 6 at 37° C. and 5% CO ₂ incubated for hours. To evaluate the release of DOX from PTS-DP during light irradiation, cells were light-irradiated using a 670 nm laser ^{at 100 mW cm -2 for 2 min.} After incubation, the medium was aspirated, washed 3 times with 1x DPBS, fixed with 4% paraformaldehyde (PFA), and 4',6-diamidino-2-phenylindole (4'amidino-2- phenylindole, DAPI). 75 nM of Lysotracker DND-26 Green in 200 μl of live cell imaging solution was added and incubated for 15 min. After 15 minutes of incubation, cells were washed 3 times with 1×DPBS and counterstained with ^{Hoechst 33342 (1 μg ml −1 ) for 15 minutes.} After incubation, cells were washed again with 1x DPBS and then supplemented with live cell imaging solution. To visualize the fluorescence obtained from the sample, confocal laser scanning microscopy (CLSM) was used. After the cells were treated with PTS-DP, cellular uptake and subsequent intracellular release of PTS-DP nanomicelles were evaluated, and then 1% sodium dodecyl sulfate (SDS) in DMSO was used to extract DOX and PhA. cells were lysed with The fluorescence intensities of DOX and PhA were measured (DOX: emission = 550 nm, excitation = 470 nm/PhA: excitation = 415 nm and emission = 673 nm).

6. Measurement of Intracellular ROS Accumulation Characteristics

To measure intracellular ROS, 2''dichlorodihydrofluorescein diacetate (DCF-DA) was used as a fluorescent probe. Briefly, ^{after seeding 1 ×10 5} CT-26 cells ml −1 and incubating with samples and laser irradiation for different time intervals, the cells were treated with 25 μM DCF-DA in D-Hanks solution and further for 20 min. incubated. After incubation, the fluorescence spectra of the cells were recorded at an excitation wavelength of 434 nm and an emission wavelength of 535 nm.

7. In vivo distribution measurement

All animal experiments were performed in accordance with the guidelines of Chonnam National University Medical School and Chonnam National University Hospital (CNU IACUC-H-2018-59) in Korea. 5-week-old male inbred Balb/c nude mice (Orient Bio Inc., Seongnam-si, Korea) were laterally subcutaneously injected with ^{1.0 × 10 6 CT-26 cells.} When the tumor volume reached approximately 100 mm 3 , mice were randomized and used for biodistribution studies. For this experiment, PhA alone and PTS-DP (10 mg kg ⁻¹ PhA) were injected intravenously (iv). The biodistribution and tumor accumulation profiles of PTS-DP were estimated from optical images at different time intervals using a fluorescence-labeled organism bio-imaging instrument (FOBI; Neo-Science, Gyeonggi-do, Korea). The tumor accumulation potential of PTS-DP was evaluated by measuring the near infrared fluorescence (NIRF) intensity. Mice were sacrificed 24 hours after injection and all major organs and tumors were collected. NIRF images of dissected normal organs (such as liver, lung, spleen, heart and kidney) and tumors were evaluated.

8. Singlet oxygen generation (SOG) in vivo

SOG in vivo was monitored using DCF-DA. PTS-DP ( ^{dose corresponding to 5 mg kg −1 of} DOX and 2.5 mg kg ⁻¹ of PhA) was injected intravenously into CT-26 tumor bearing mice. After 24 hours, DCF-DA (50 mg kg ⁻¹ ) was injected intratumorally and irradiated with a 670 nm laser (100 mW cm ⁻² ) for 15 minutes. Tumor tissues were collected and frozen sections (6 μm) were performed and counterstained with DAPI.

9. Chemo-photodynamic therapy for tumor-bearing mice

CT-26 tumor bearing mice with tumor volumes between 80 and 100 mm 3 were used for chemo-photodynamic studies. Tumor bearing mice were divided into 8 groups with 4 mice in each group. (8 groups treated with PBS, DOX, PhA, PTS-DP, PBS + laser, DOX + laser, PhA + laser and PTS-DP + laser) Control and sample groups were injected intravenously. DOX at doses corresponding to 5 mg kg ⁻¹ and 2.5 mg kg ⁻¹ of PhA were injected twice with an interval of 7 days, and after 24 h, the tumor area was exposed to 670 nm light of ^{100 mW cm −2 for 15 min.} Tumor volume and whole body weight of animals were measured every 2 days.

[Example 1] Preparation of polymer micelles using an amphiphilic polymer compound

As in FIG. 2, polyethylene glycol-TL-stearamine (PTS) is prepared through the process of steps 1 to 3.

Step 1: Preparation of thioketal linker (TL)

49 mmol of 3-mercaptopropionic acid was dissolved in 98 mmol of anhydrous acetone, and the resulting product was cooled to crystallization while constantly stirring at 23 °C for 6 hours. The crystallized product was filtered, rinsed with n-hexane, and then rinsed again with cold distilled water, and then freeze-dried.

Step 2: Synthesis of thioketallinker-conjugated polyethylene glycol (PEG-TL)

254 mg of the thioketal linker (TL) prepared in step 1 and 200 mg of methoxy-polyethylene glycol amine (PEG-AM) having a molecular weight of 2 kDa were mixed with 10 ml of N,N-dimethylformamide (N,N-Dimethylformamide) , DMF) and mixed at room temperature to prepare a mixed solution. Then, 278 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, EDC] and 345 mg of N-hydroxyl After the addition of succinimide (N-hydroxysuccinimide, NHS), the mixture was stirred for 16 hours. After stirring, the mixed solution was dialyzed against distilled water under a condition of a molecular cut-off (MWCO) of 1,000 to remove reaction by-products and then freeze-dried. The freeze-dried powder was redissolved in 1 ml DMF, and precipitated in cold diethyl ether 5 times repeatedly. Thereafter, by vacuum drying, polyethylene glycol (PEG-TL) conjugated with a thioketal linker was prepared.

Step 3: Polyethylene glycol-TL-stearamine (PTS) production

100 mg of PEG-TL prepared in step 2, 80 ml of triethylamine (TEA) and 80 mg of stearamine (C18) were mixed with 10 ml of N,N-dimethylformamide (N,N- dimethylformamide, DMF) and mixed at 80 °C. To the thoroughly mixed solution, 240 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, EDC] and 145 mg of hydroxysuccin were added. Mid (N-hydroxysuccinimide, NHS) was added and stirred for one day. After the reaction, the solution was purified by dialysis against distilled water for 2 days at a molecular cut-off (MWCO) of 2,000, and then freeze-dried to prepare polyethylene glycol-TL-stearamine (PTS).

For polymer micelles prepared from PTS, aqueous solutions containing polymer micelles of various concentrations were prepared to measure critical micelle concentration (CMC). As the fluorescent probe, 6.0×10 ^-7 M of pyrene was used, and the excitation wavelength was set to 336 nm in a fluorescence spectrophotometer, and the emission wavelength was observed in the range of 360-450 nm. As a result of the measurement, the CMC of the PTS polymer micelles was 0.2 mg/mL.

In order to confirm the formation of polymer micelles, a solvent in which PTS was self-assembled in a D2O solvent was prepared, and then 1H-NMR of a D2O solution of PTS was measured. PTS was completely shielded in D2O as the stearic group formed a hydrophobic core in the D2O solvent, and peaks at δ=0.83 and 1.23, which are peaks caused by the alkylene group, almost disappeared. The above results suggest that PEG formed a hydrophilic shell and a stearic group formed a hydrophobic core to form a polymeric micellar.

[Example 2] Preparation of a drug delivery system comprising a photodynamic compound in the hydrophobic core of the polymer micelle

1 mg of pheophorbide A (PhA) was mixed with 10 mg of PTS prepared in Example 1 in chloroform (oil phase). The oil phase was then added dropwise to 10 ml of distilled water (aqueous phase) under sonication. The solution was stirred in the dark for 24 hours, then dialyzed against distilled water under the condition of molecular cut-off (MWCO) of 10-12 kDa, and then freeze-dried to PTS drug containing pheophorbide A (PhA) inside. A delivery vehicle (PTS-P1) was prepared.

[Example 3] Preparation of a drug delivery system comprising a photodynamic compound in the hydrophobic core of the polymer micelle

It was prepared in the same manner as in Example 2, except that 2 mg of pheoforbid A (PhA) was used to form a PTS drug delivery system (PTS-P2).

[Example 4] Preparation of a drug delivery system containing a hydrophobic drug in the hydrophobic core of the polymer micelle

First, triethylamine (TEA) was added to a mixed solution of N,N-dimethylformamide (N,N-Dimethylformamide, DMF) and doxorubicin-hydrochloride (DOX-HCl) and hydrophobic doxorubicin (DOX). ) was formed. Then, 1 mg of doxorubicin was mixed with 10 mg of the PTS prepared in Example 1 in chloroform (oil phase). The oil phase was then added dropwise to 10 ml of distilled water (aqueous phase) under sonication. The solution was stirred in the dark for 24 hours, then dialyzed against distilled water under the condition of molecular cut-off (MWCO) of 10-12 kDa and freeze-dried to PTS drug delivery system (PTS) containing doxorubicin (DOX) inside. -D1) was formed.

[Example 5] Preparation of a drug delivery system comprising a hydrophobic drug in the hydrophobic core of the polymer micelle

A PTS drug delivery system (PTS-D2) was formed in the same manner as in Example 4, except that 2 mg of doxorubicin (DOX) was used.

[Example 6] Preparation of a drug delivery system comprising a hydrophobic drug and a photodynamic compound in the hydrophobic core of the polymer micelle (hydrophobic drug: photodynamic compound = 1:1)

First, triethylamine (TEA) was added to a mixed solution of N,N-dimethylformamide (N,N-Dimethylformamide, DMF) and doxorubicin hydrochloride (DOX-HCl), followed by hydrophobic doxorubicin (Doxorubicin). , DOX) was formed. Then, 1 mg of doxorubicin and 1 mg of pheophorbide A (PhA) were mixed with 10 mg of the PTS prepared in Example 1 in chloroform (oil phase). The oil phase was then added dropwise to 10 ml of distilled water (aqueous phase) under sonication. The solution was stirred in the dark for 24 hours, and then dialyzed against distilled water under the condition of molecular cut-off (MWCO) of 10-12 kDa and freeze-dried to insert doxorubicin (DOX) and pheophorbide A (PhA) inside. A PTS drug delivery system [PTS-DP (1:1)] contained in was formed.

[Example 7] Preparation of a drug delivery system comprising a hydrophobic drug and a photodynamic compound in the hydrophobic core of the polymer micelle (hydrophobic drug: photodynamic compound = 2: 1)

PTS drug delivery system [PTS-DP (2:1)] was formed in the same manner as in Example 6, except that 2 mg of doxorubicin (DOX) and 1 mg of pheophorbide A (PhA) were used. did.

[Example 8] Preparation of a drug delivery system comprising a hydrophobic drug and a photodynamic compound in the hydrophobic core of the polymer micelle (hydrophobic drug: photodynamic compound = 1: 2)

PTS drug delivery system [PTS-DP (1:2)] was formed in the same manner as in Example 6 except that 1 mg of doxorubicin (DOX) and 2 mg of pheophorbide A (PhA) were used. did.

Characteristics were evaluated for the above examples as follows.

[Experimental Example 1] Confirmation of production of an amphiphilic high molecular compound

1H-NMR spectrum and FT-IR spectrum measured using 1H-NMR and FT-IR spectroscopy to confirm the composition of the PTS prepared in Example 1 are shown in FIGS. 3A and 3B, respectively.

[Experimental Example 2] Composition ratio, hydrodynamic diameter and zeta potential of polymer micelles

The drug content, content and drug encapsulation efficiency of micelles prepared in Examples 1 to 8 are shown in Table 1 below.

content
(Unit: mg) Content rate of active ingredient
(unit: %) Encapsulation Efficiency of Active Ingredients
(unit: %) Michelle DOX PhA DOX PhA DOX PhA Example 1 PTS 0 0 - - - - Example 2 PTS-P1 0 One 0 4.75±0.02 0 52.26±0.18 Example 3 PTS-P2 0 2 0 17.04±0.11 0 83.23±1.86 Example 4 PTS-D1 One 0 3.78±0.02 0 41.58±2.23 0 Example 5 PTS-D2 2 0 7.09±0.23 0 42.52±1.40 0 Example 6 PTS-DP(1:1) One One 17.24±0.53 13.43±2.04 85±1.06 67.1±4.08 Example 7 PTS-DP (2:1) 2 One 16.94±0.41 11.36±0.09 86.38±4.46 68.14±0.59 Example 8 PTS-DP(1:2) One 2 17.48±0.31 17.30±0.04 88.91±3.91 80.34±0.50

The average hydrodynamic diameter and zeta potential of the micelles prepared in Examples 1 to 8 are shown in FIG. 4A, and the FE-TEM image of the micelles prepared in Example 6 is shown in FIG. 4B.

As can be seen in Table 1 and FIG. 4, when DOX and PhA are included as active ingredients inside the PTS micelles, the hydrodynamic diameter is higher than when no active ingredients are included or DOX or PhA is included as a single species. It can be seen that this has a small particle size of 80 to 200 nm. In addition, the PTS-DP (1:1) of Example 6 containing the active ingredients DOX and PhA in a weight ratio of 1:1 showed excellent active ingredient encapsulation efficiency, and the hydrodynamic diameter measured using DLS analysis was about 85 It turned out that it has a small particle diameter of nanometer. This appears to be an effect due to the presence of π-π stacking between doxorubicin (DOX) and pheophorbide (PhA).

[Experimental Example 3] Measurement of release of active ingredient of drug delivery system

In order to measure the emission characteristics of DOX and PhA of the PTS-DP prepared in Example 6, light of a wavelength of 670 nm was irradiated with 100 mW cm ^-2 intensity for 30 minutes, and 100 μM H ₂ O similar to the intracellular ROS environment. _{Maintaining 2} concentration conditions, 1 ml of release medium was collected at different time intervals.

Then, the concentration of DOX in the release medium was quantified by high performance liquid chromatography (HPLC) analysis, and hydrophobic PhA was extracted using dichloromethane (DCM), and the concentration of PhA was measured through HPLC analysis.

The quantification of DOX and PhA emitted while changing the ambient conditions of the PTS-DP using high performance liquid chromatography (HPLC) is shown in FIG. 5 .

As shown in FIGS. 5A and 5B , it can be seen that the emission of DOX and PhA is higher when irradiated with light compared to the non-irradiated condition. That is, it can be seen that the additional light irradiation increases the concentration of ROS to increase the initial release properties of DOX and PhA. Moreover, it can be seen that it has a more rapid release of DOX and PhA when irradiated with light under _{H 2} O _{2 conditions.} That is, the drug delivery system of the present invention has excellent DOX and PhA release properties in cancer cell conditions, and thus can be effectively used as a chemo-photodynamic therapeutic agent.

[Experimental Example 4] Measurement of photodynamic therapy and anticancer effect of drug delivery system in vitro

Cell viability (% of cell viability), ROS production rate, and apoptosis during light irradiation of the PTS-DP prepared in Example 6 and the control were measured.

Figure 6a shows the determination of cell viability using the CCK-8 assay after culturing the PTS-DP and controls prepared in Example 6 with CT26 cells, respectively, in order to measure the apoptosis effect of CT26 cells, Figure 6b shows the quantification of intracellular ROS using the fluorescent probe DCFDA to confirm the effective generation of ROS by the release of PhA inside the drug delivery system. 6c and 6d show the measurement of the production rate of caspase 3, an apoptosis factor.

It can be seen that when light irradiation is applied to the PTS-DP as in FIG. 6b, the generated ROS is more improved compared to the control groups, and in FIG. have. Moreover, it can be seen that the generation of Caspase 3, a factor involved in apoptosis, was significantly higher than that of the control group in FIGS. 6c and 6d . This is to increase the amount of ROS production by the rapid decomposition of PTS-DP upon light irradiation, thereby inducing the release of a greater amount of DOX to increase apoptosis, confirming that it can be used as an effective anti-cancer treatment method.

[Experimental Example 5] Measurement of biodistribution and biocompatibility

The cell accumulation rate and biocompatibility of the PTS-DP prepared in Example 6 and the control were measured.

Figure 7a shows the in vivo distribution of the PTS-DP prepared in Example 6 by injection into a vein of a CT26 tumor-bearing mouse, and Figure 7b shows the in vitro fluorescence imaging of the tumor. FIG. 7c shows the change in concentration with time after PTS-DP injection, and FIG. 7d shows the concentration of DOX by light irradiation of PTS-DP.

As shown in FIGS. 7A and 7B , it can be seen that PTS-DP has an excellent accumulation rate for tumors. It can be seen that PTS-DP has excellent in vivo circulation stability, improved permeability, and a higher accumulation rate in tumors than in normal cells.

In addition, it can be seen in FIG. 7c that PTS-DP has a longer circulation time in blood than DOX, a control, and in FIG. 7d, when light irradiation is concurrently with PTS-DP, the decomposition of PTS-DP proceeds rapidly and DOX is released in large amount, it can be seen that the DOX accumulation rate in the tumor increases. That is, the PTS-DP prepared according to one aspect of the present invention has a long circulation time in the blood, can implement a high accumulation rate of selective target cells, and has rapid drug release by light irradiation, so that it is an effective drug delivery system. And it was confirmed that it has a function as a therapeutic agent.

[Experimental Example 6] Suitability of chemo-photodynamic therapy

In order to confirm the suitability of the chemo-photodynamic treatment method of PTS-DP prepared in Example 6, rats with CT26 tumor were randomly grouped into 8 groups of 4 each, and PBS, DOX, PhA and PTS-DP were Each was injected intravenously. Of these, only four groups ^{were irradiated with light under conditions of 670 nm and 100 mW cm -2} for 15 minutes, and the volume change of the tumor was monitored for 18 days, as shown in FIG. 8a. In addition, on day 18 after injection, tumors were collected from all groups of mice, and the average tumor weight was measured and shown in FIG. 8B .

^{9 shows the generation of singlet oxygen ( 1} O ₂ ) before and after light irradiation on PTS-DP in CT-26 tumor cells by fluorescence microscopy images.

As shown in FIG. 8a , it can be seen that when PTS-DP and light irradiation were combined, the tumor volume was reduced by about 3 times or more compared to other groups. This shows the effect of effectively inhibiting the growth of tumor cells due to the rapid and local release of ROS and DOX by the degradation of ROS and PTS-DP by light irradiation of the tumor. In addition, in FIG. 8b , in the absence of light irradiation, the tumor weights of the groups injected with PTS-DP and other groups did not differ significantly, but when light irradiation was concurrently performed, the tumor weight was reduced by about 3 times compared to before light irradiation. could confirm that

It can be seen from FIG. 9 that the generation of ROS of PhA is increased by light irradiation, which is an effect due to accelerated decay of PTS-DP. Moreover, it was found that the toxicity was low through the small change in body weight of the rats of the PTS-DP injection group.

From the above experimental results, the micelles formed of the amphiphilic polymer compound of the present invention and the drug delivery system carrying the active ingredients in the micelles can maintain a stable structure even in the blood, which is a living condition, and the encapsulation efficiency of the active substances is significantly higher than that of the control group. As well as excellent, it was confirmed that the local accumulation rate in the target cell was excellent. Furthermore, the drug delivery system of the present invention is a treatment combining photodynamic therapy using light irradiation and chemotherapy using anticancer agents such as doxorubicin (DOX), and minimizes toxic side effects. It was confirmed that the cell growth inhibition was induced.

From this, it was confirmed that micelle containing a drug and a photodynamic compound inside using the amphiphilic polymer compound of the present invention has an excellent effect as a drug delivery system with excellent targeting of target cancer cells, and a tumor treatment system using the same is an effective anticancer It was also confirmed to have a therapeutic effect.

As described above, the present invention has been described by specific matters and limited examples, but these are only provided to help a more general understanding of the present invention, and the present invention is not limited to the above examples, and the field to which the present invention belongs Various modifications and variations are possible from these descriptions by those of ordinary skill in the art.

Therefore, the spirit of the present invention should not be limited to the described embodiments, and not only the claims to be described later, but also all those with equivalent or equivalent modifications to the claims will be said to belong to the scope of the spirit of the present invention. .

Claims (13)

An amphiphilic polymer compound wherein one end of the hydrophilic water-soluble polymer and one end of the compound having a hydrophobic hydrocarbon group are covalently bonded with a thioketal linker (TL). According to claim 1,
The hydrophilic water-soluble polymer is polyethylene glycol, acrylic acid-based polymer, methacrylic acid-based polymer, polyamidoamine, poly(2-hydroxypropyl methacrylamide)-based polymer, water-soluble polypeptide, water-soluble polysaccharide or polyglycerol, an amphiphilic polymer compound . According to claim 1,
The hydrocarbon group of the compound having a hydrophobic hydrocarbon group is an aliphatic hydrocarbon group of (C12 to C22), an amphiphilic high molecular compound. According to claim 1,
The ratio of the weight average molecular weight of the hydrophilic water-soluble polymer to the molecular weight of the compound having a hydrophobic hydrocarbon group satisfies Formula 1 below.
[Equation 1]

(In Formula 1, WSP means the weight average molecular weight of the hydrophilic water-soluble polymer, and HC means the molecular weight of the compound having a hydrophobic hydrocarbon group) According to claim 1,
One end of the hydrophilic water-soluble polymer and one end of the compound having a hydrophobic hydrocarbon group are each covalently bonded to a thioketal linker with an amide group, an amphiphilic polymer compound. According to claim 1,
The amphiphilic polymer compound is polyethylene glycol-TL-(C12~C22)alkylamine. A polymer micelle formed by self-assembly of the amphiphilic polymer compound of any one of claims 1 to 6 in an aqueous phase. 8. The method of claim 7,
The polymer micelle is a drug delivery system comprising a hydrophobic drug and a photodynamic compound in a hydrophobic core. 9. The method of claim 8,
The hydrophobic drug is a hydrophobic anticancer compound, and the thioketal linker has a reactivity by reactive oxygen species, a drug delivery system. 9. The method of claim 8,
The photodynamic compound is any one or two or more selected from the group consisting of a porphyrin-based compound, a chlorine-based compound, a bacteriochlorine-based compound, a phthalocyanine-based compound, a naphthalocyanine-based compound, and a 5-aminolevulin-based compound Phosphorus, drug delivery system. 9. The method of claim 8,
The drug delivery system reacts to reactive oxygen species around the cell membrane and primarily releases a portion of the hydrophobic drug, and reacts to the reactive oxygen species generated by light irradiation to collapse the polymer micelle to release the hydrophobic drug contained in the hydrophobic core 2 A drug delivery system, characterized in that it releases all of the secondary. A tumor treatment system comprising a near-infrared irradiator and the drug delivery system according to claim 8. 13. The method of claim 12,
The light irradiated from the near-infrared irradiator is absorbed by the photodynamic compound included in the drug delivery system to generate reactive oxygen species, a tumor treatment system.

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