Pharmaceutical composition for photodynamic therapy of cancer comprising nitrobenzene-amino acid-metal ion-based nanocomposite as active ingredient

Abstract

The present invention relates to a pharmaceutical composition for photodynamic therapy of cancer, comprising a nitrobenzene-amino acid-metal ion-based nanocomposite as an active ingredient. The nanocomposite specifically reacts with glutathione in cancer cells to enable near-infrared imaging, and has been confirmed to have an anticancer effect by reactive oxygen species generated by light irradiation, and thus can be effectively used in related industries.

Description

Pharmaceutical composition for photodynamic therapy of cancer comprising a nitrobenzene-amino acid-metal ion-based nanocomposite as an active ingredient

[0001] The present invention relates to a pharmaceutical composition for photodynamic therapy of cancer comprising a nitrobenzene-amino acid-metal ion-based nanocomplex as an active ingredient.

[0002] Bioactive substances possess special abilities, such as enzyme-like behavior, structural similarity, and variable cell signaling regulation, expanding the scope of research by bridging basic science and clinical practice. The development of biomolecules into bioactive substances demonstrates high biocompatibility for clinical use.

[0003] When selecting elements for the design of bioactive compounds, thiol-containing compounds have been considered promising due to their ease of manipulation via click chemistry and their bioorthogonal affinity. Among these, thiol-containing compounds, such as cysteine ​​(Cys), homocysteine ​​(Hcy), and glutathione (GSH), have been extensively studied to reveal their antioxidant, antimicrobial, and anticancer properties through processes that reduce reactive oxygen species (ROS). Consequently, the overexpression of thiols in the tumor microenvironment has been confirmed, underscoring the imperative for the development of thiol-activated therapeutic agents with tumor-specific selectivity.

[0004] To date, biocompatible metal-coordination compounds have been shown to outperform non-metallic compounds in the tumor microenvironment through a glutathione-regulated redox system, and to enhance the efficacy of chemodynamic therapy (CDT) against cancer by generating reactive oxygen species. However, the oxidation of metal ions remains challenging due to the relatively low concentration of hydrogen peroxide within tumors. Furthermore, the lack of a tumor monitoring system and the insufficient production of reactive oxygen species pose several challenges. Therefore, current research is focused on developing systems that increase reactive oxygen species.

[0005] Photodynamic therapy (PDT) is a treatment method that generates reactive oxygen species (ROS) through light activation of a photosensitizer. This method can enhance the production of ROS and offers the advantage of precise targeting of specific areas. Therefore, PDT can enhance ROS production and complement chemodynamic therapy.

[0006] Accordingly, the inventors of the present invention have completed the present invention by developing an anticancer agent capable of tumor-specific near-infrared imaging, chemodynamic therapy based on the Fenton reaction, and photodynamic therapy by utilizing the specificity of sulfide substances such as glutathione of nitrobenzene-amino acid-metal nanoparticles.

[0007] This patent was supported by the Ministry of Science and ICT (National Research Foundation of Korea) (Project No. 1711192122, Subproject No. 2022R1F1A1069954).

[0008] This patent was supported by the Ministry of Education (National Research Foundation of Korea) (Project No.: 1345365662, Subproject No.: 2018R1A6A1A03025124).

[0009] This patent was supported by the Ministry of Science and ICT (Science and ICT Commercialization Promotion Agency) (RS-2024-00418924).

[0010] The purpose of the present invention is to provide a nanocomposite represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0011] [Chemical Formula 1]

[0012]

[0013] (In the above chemical formula 1,

[0014] A is oxygen (O) or selenium (Se),

[0015] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy,

[0016] M is a divalent cation metal.)

[0017] Another object of the present invention is to provide a pharmaceutical composition for photodynamic therapy of cancer comprising a nanocomposite represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0018] Another object of the present invention is to provide a method for producing a nanocomposite produced by the following reaction scheme 1, which comprises the steps of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent and adding copper cations to the reactants.

[0019] [Reaction Formula 1]

[0020]

[0021] Another object of the present invention is to provide a near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the above chemical formula 1.

[0022] Another object of the present invention is to provide a method for detecting intracellular glutathione, comprising the step of adding a nanocomplex represented by the above chemical formula 1 to a sample.

[0023] Another object of the present invention is to provide a nanocomposite represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0024] [Chemical Formula 2]

[0025]

[0026] (In the above chemical formula 2,

[0027] A is oxygen (O) or selenium (Se),

[0028] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy,

[0029] M is a divalent cation metal.)

[0030] Another object of the present invention is to provide a method for producing a nanocomposite produced by the following reaction scheme 2, which comprises the steps of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent and adding copper cations to the reactants.

[0031] [Reaction Formula 2]

[0032]

[0033] Another object of the present invention is to provide a near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the above chemical formula 2.

[0034] Another object of the present invention is to provide a method for detecting intracellular glutathione, comprising a step of adding a nanocomplex represented by the above chemical formula 2 to a sample.

[0035] Another object of the present invention is to provide a method for photodynamic treatment of cancer, comprising the step of administering to a subject a pharmaceutically effective amount of a nanocomposite represented by the above chemical formula 1.

[0036] To achieve the above purpose,

[0037] The present invention provides a nanocomposite represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0038] [Chemical Formula 1]

[0039]

[0040] (In the above chemical formula 1,

[0041] A is oxygen (O) or selenium (Se),

[0042] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy,

[0043] M is a divalent cation metal.)

[0044] In addition, the present invention provides a pharmaceutical composition for photodynamic therapy of cancer, comprising a nanocomposite represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0045] In addition, the present invention provides a method for producing a nanocomposite produced by the following reaction scheme 1, which comprises the step of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent and the step of adding copper cations to the reactants.

[0046] [Reaction Formula 1]

[0047]

[0048] In addition, the present invention provides a near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the above chemical formula 1.

[0049] In addition, the present invention provides a method for detecting intracellular glutathione, comprising the step of adding a nanocomplex represented by the above chemical formula 1 to a sample.

[0050] In addition, the present invention provides a nanocomposite represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0051] [Chemical Formula 2]

[0052]

[0053] (In the above chemical formula 2,

[0054] A is oxygen (O) or selenium (Se),

[0055] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy,

[0056] M is a divalent cation metal.)

[0057] In addition, the present invention provides a method for producing a nanocomposite produced by the following reaction scheme 2, which comprises the step of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent and the step of adding copper cations to the reactants.

[0058] [Reaction Formula 2]

[0059]

[0060] In addition, the present invention provides a near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the above chemical formula 2.

[0061] In addition, the present invention provides a method for detecting intracellular glutathione, comprising a step of adding a nanocomplex represented by the above chemical formula 2 to a sample.

[0062] In addition, the present invention provides a method for photodynamic treatment of cancer, comprising the step of administering to a subject a pharmaceutically effective amount of a nanocomposite represented by the above chemical formula 1.

[0063] Novel nanocomposite NPS-Cys-Cu of the present invention 2+ It reacts specifically with glutathione in cancer cells, enabling near-infrared imaging, and its anticancer effect by reactive oxygen species generated by light irradiation has been confirmed, so it can be usefully utilized in related industries.

[0064] Figure 1 shows an FT-IR spectrum of Fourier transform infrared spectroscopy proving the number of coordination bonds of the complex of the present invention.

[0065] Figure 2 shows the fluorescence selectivity of the complex of the present invention toward biothiol, biomolecule or metal ion.

[0066] Figure 3 shows fluorescence photographs and fluorescence emission spectra of the complex of the present invention before and after glutathione reaction in a 365 nm UV chamber.

[0067] Figure 4 shows the absorption spectrum of reactive oxygen species generated in the presence of hydrogen peroxide or when irradiated with light having a wavelength of 532 nm in the complex of the present invention.

[0068] Figure 5 shows the results of electron paramagnetic resonance analysis of reactive oxygen species generated in the presence of hydrogen peroxide or when the complex of the present invention is irradiated with light at a wavelength of 532 nm.

[0069] Figure 6 shows the fluorescence on-off phenomenon of cells according to the intracellular glutathione concentration of the complex of the present invention.

[0070] Figure 7 shows the intracellular production of reactive oxygen species by the complex of the present invention.

[0071] Figure 8 shows the therapeutic effect of complex 1 injected subcutaneously into tumor cell line transplanted mice with or without photoirradiation, as shown in the tumor size and weight of the mouse.

[0072] Figure 9 shows the angiogenic factor formation level of a tumor treated by administering the complex of the present invention.

[0073] Figure 10 illustrates the entire process of the complex of the present invention being activated within cancer tissue to treat cancer.

[0074] Hereinafter, the present invention will be described in detail with reference to the attached drawings and embodiments thereof. However, the following embodiments are provided as illustrative examples of the present invention. If a detailed description of a technology or configuration well known to those skilled in the art is judged to unnecessarily obscure the gist of the present invention, such detailed description may be omitted, and the present invention is not limited thereby. The present invention is capable of various modifications and applications within the scope of the following claims and equivalents interpreted therefrom.

[0075] In addition, the terminology used in this specification is a term used to appropriately express preferred embodiments of the present invention, and this may vary depending on the intention of the user or operator, or the customs of the field to which the present invention belongs. Therefore, the definition of these terms should be determined based on the contents throughout this specification. Throughout the specification, when a part is said to "include" a certain component, this does not mean that other components are excluded, but rather that other components may be included, unless specifically stated otherwise.

[0076] Hereinafter, terms used in the present invention are explained.

[0077] The present invention provides a nanocomposite represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0078] [Chemical Formula 1]

[0079]

[0080] (In the above chemical formula 1,

[0081] A is oxygen (O) or selenium (Se),

[0082] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy,

[0083] M is a divalent cation metal.)

[0084] The "complex", "complex" or "coordination compound" of the present invention is a substance in which several other substances form a single atomic group by forming a three-dimensional coordinate covalent bond around one atom or ion in a directional manner, and the lone pair of electrons of the ligand is formed through a coordination bond to a central metal ion having many empty orbitals.

[0085] The above "coordinate bond" refers to a case where, when two atoms form a covalent bond, the electrons involved in the bond are formally donated by only one atom. The distinction between a normal covalent bond and a coordinate bond is an artificial one, but it is often used in books, etc. In addition, the explanation of the bonding force and bonding is the same as for other polar covalent bonds. A coordinate bond is formed when a Lewis base (electron pair donor) donates an electron pair to a Lewis acid (electron pair acceptor). When the electron pair acceptor receives a negative charge, the electron pair donor receives a positive charge.

[0086] According to one embodiment of the present invention, the metal of the coordination bond may include any metal having a divalent cation, and may preferably be copper.

[0087] According to one embodiment of the present invention, the nanocomposite may be the following complex.

[0088]

[0089] The above complex may be named nitrobenzoselenadiazole-cysteine-copper (hereinafter, “complex 1”).

[0090] According to one embodiment of the present invention, the complex may specifically react with glutathione.

[0091] The "Glutathione (GSH)" of the present invention is a crystalline peptide composed of three amino acids: glutamic acid, cysteine, and glycine. It is an antioxidant found in plants, animals, fungi, and some bacteria and archaea. Glutathione can prevent damage to important cellular components caused by reactive oxygen species, such as free radicals, superoxide dismutases, lipid peroxides, and heavy metals.

[0092] According to one embodiment of the present invention, the complex may react with glutathione to form a compound represented by the following chemical formula 3.

[0093] [Chemical Formula 3]

[0094]

[0095] (In the above chemical formula 3,

[0096] A is oxygen (O) or selenium (Se),

[0097] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, or alkoxy.

[0098] The "compound" of the present invention is a pure chemical substance formed by combining atoms of two or more types of chemical elements, and can be separated into simpler substances through a chemical reaction.

[0099] According to one embodiment of the present invention, the compound may be the following compound.

[0100]

[0101] The above compound may be named nitrobenzoselenadiazole-cysteine ​​(hereinafter, “compound 1”).

[0102] In the present invention, the compound can be used as a fluorescent substance.

[0103] The "phosphor" or "fluorophore" of the present invention is a material that exhibits a luminescent phenomenon. It emits light when exposed to some type of radiant energy. This includes fluorescent or phosphorescent materials that emit light when exposed to ultraviolet or visible light, and cathode luminescent materials that emit light when struck by an electron beam in a cathode ray tube.

[0104] According to one embodiment of the present invention, the compound may emit fluorescence at 400 to 700 nm.

[0105] According to one embodiment of the present invention, the nanocomposite may react with glutathione to reduce a divalent cation metal to a monovalent cation metal.

[0106] According to one embodiment of the present invention, the monovalent cationic metal may react with a biomolecule to generate reactive oxygen species.

[0107] The "reactive oxygen species (ROS)" or "active oxygen species" of the present invention are chemically reactive molecules containing oxygen atoms. They are compounds produced within living organisms and are highly oxidizing oxygen species that can attack biological tissues and damage cells. Reactive oxygen species are produced during normal cellular activity and are involved in various biological processes, including cell differentiation, gene expression, and cytokine responses.

[0108] According to one embodiment of the present invention, the biomolecule may include an active substance including hydrogen peroxide, an amino acid, a protein, a lipid, a byproduct of cell metabolism, or oxygen.

[0109] According to one embodiment of the present invention, the reactive oxygen species may be, but is not limited to, at least one selected from the group consisting of a hydroxyl radical, singlet oxygen, hydrogen peroxide, superoxide, or nitric oxide, and more specifically, may be a hydroxyl radical or singlet oxygen.

[0110] According to one embodiment of the present invention, the compound may increase intracellular Endostatin / collagen XVIII, MMP-3, Coagulation Facter III, IL-1a, PEDF, IL-1b or CD26 protein expression.

[0111] According to one embodiment of the present invention, the compound may be photo-irradiated to reduce intracellular Endostatin / collagen XVIII, MMP-3, Coagulation Factor III, IL-1a, PEDF, IL-1b or CD26 protein expression.

[0112] In addition, the present invention provides a pharmaceutical composition for photodynamic therapy of cancer, comprising a nanocomposite represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0113] The pharmaceutical composition of the present invention may further include an adjuvant in addition to the active ingredient. Any adjuvant known in the art may be used without limitation. However, for example, Freund's complete adjuvant or incomplete adjuvant may be further included to enhance its effectiveness.

[0114] The pharmaceutical composition according to the present invention can be prepared in a form in which the active ingredient is mixed with a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients, and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers that can be used in the pharmaceutical composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0115] The pharmaceutical composition of the present invention can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions, each according to a conventional method.

[0116] When formulated, it can be prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and such solid preparations can be prepared by mixing the active ingredient with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives can be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, Tween 61, cocoa butter, laurin, and glycerogelatin.

[0117] The pharmaceutical composition according to the present invention can be administered to a subject via various routes. All modes of administration are contemplated, including oral, intravenous, intramuscular, subcutaneous, and intraperitoneal injection.

[0118] The dosage of the pharmaceutical composition according to the present invention is selected in consideration of the age, weight, sex, physical condition, etc. of the subject. It is obvious that the concentration of the active ingredient included in the pharmaceutical composition can be selected in various ways depending on the subject, and it is preferably included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg / ml. If the concentration is less than 0.01 μg / ml, pharmaceutical activity may not be observed, and if it exceeds 5,000 μg / ml, it may be toxic to the human body.

[0119] According to one embodiment of the present invention, the cancer may be at least one selected from the group consisting of brain cancer, spinal cancer, lung cancer, thyroid cancer, breast cancer, skin cancer, prostate cancer, and ovarian cancer, but is not limited thereto, and more specifically, may be brain cancer.

[0120] In the present invention, the brain cancer may include malignant glioma. The malignant glioma is a malignant tumor that arises from cells that make up the brain or spinal cord.

[0121] According to one embodiment of the present invention, the composition may inhibit angiogenesis of cancer.

[0122] The "angiogenesis" of the present invention refers to cancer angiogenesis, a process in which substances that induce angiogenesis are secreted around cancer cells or tissues to supply oxygen necessary for growth according to the rapid growth rate of cancer cells. Due to the formed new blood vessels, cancer can grow rapidly again, and this process is repeated until the cancer disappears.

[0123] In addition, the present invention provides a method for producing a nanocomposite produced by the following reaction scheme 1, which comprises the step of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent and the step of adding copper cations to the reactants.

[0124] [Reaction Formula 1]

[0125]

[0126] According to one embodiment of the present invention, the organic solvent may be any one selected from the group consisting of tetrahydrofuran (THF), dimethylformamide (DMF), dichloromethane (CH2Cl2), 1,2-dichloroethane (CH2ClCH2Cl), dimethyl sulfoxide (DMSO), t-butyl methyl ether (TBME), acetonitrile (ACN), methanol, ethanol, isopropanol, t-butanol, diethyl ether, diphenyl ether, diisopropyl ether (DIPE), dimethylacetamide (DMA), chlorobenzene, benzene, toluene, carbon tetrachloride (CCl4), acetone, trifluoroacetic acid, chloroform (CHCl3), pyridine, and aqueous solutions thereof, or a mixed solution thereof, and preferably THF.

[0127] According to one embodiment of the present invention, the reaction may be stirred at 20 to 30°C for 3 hours.

[0128] In addition, the present invention provides a near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the above chemical formula 1.

[0129] In addition, the present invention provides a method for detecting intracellular glutathione, comprising the step of adding a nanocomplex represented by the above chemical formula 1 to a sample.

[0130] In addition, the present invention provides a nanocomposite represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0131] [Chemical Formula 2]

[0132]

[0133] (In the above chemical formula 2,

[0134] A is oxygen (O) or selenium (Se),

[0135] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy,

[0136] M is a divalent cation metal.)

[0137] According to one embodiment of the present invention, the metal of the coordination bond may typically include any metal having a divalent cation, and may preferably be copper.

[0138] According to one embodiment of the present invention, the nanocomposite may be the following complex.

[0139]

[0140] The above complex may be named nitrobenzofurazan-cysteine-copper (hereinafter, “complex 2”).

[0141] According to one embodiment of the present invention, the nanocomposite may specifically react with glutathione.

[0142] According to one embodiment of the present invention, the nanocomposite may react with glutathione to form a compound represented by the following chemical formula 3.

[0143] [Chemical Formula 3]

[0144]

[0145] (In the above chemical formula 3,

[0146] A is oxygen (O) or selenium (Se),

[0147] X1 to X4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, or alkoxy.

[0148] According to one embodiment of the present invention, the compound may be the following compound.

[0149]

[0150] The above compound may be named nitrobenzofurazan-cysteine ​​(hereinafter, “compound 2”).

[0151] According to one embodiment of the present invention, the compound may emit fluorescence at 400 to 700 nm.

[0152] According to one embodiment of the present invention, the nanocomposite may react with glutathione to reduce a divalent cation metal to a monovalent cation metal.

[0153] According to one embodiment of the present invention, the monovalent cationic metal may react with a biomolecule to generate reactive oxygen species.

[0154] According to one embodiment of the present invention, the biomolecule may include an active substance including hydrogen peroxide, an amino acid, a protein, a lipid, a byproduct of cell metabolism, or oxygen.

[0155] According to one embodiment of the present invention, the reactive oxygen species may be, but is not limited to, at least one selected from the group consisting of hydroxyl radical, singlet oxygen, hydrogen peroxide, superoxide, or nitric oxide, and preferably may be a hydroxyl radical or singlet oxygen.

[0156] In addition, the present invention provides a method for producing a nanocomposite produced by the following reaction scheme 2, which comprises the step of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent and the step of adding copper cations to the reactants.

[0157] [Reaction Formula 2]

[0158]

[0159] According to one embodiment of the present invention, the organic solvent may be any one selected from the group consisting of tetrahydrofuran (THF), dimethylformamide (DMF), dichloromethane (CH2Cl2), 1,2-dichloroethane (CH2ClCH2Cl), dimethyl sulfoxide (DMSO), t-butyl methyl ether (TBME), acetonitrile (ACN), methanol, ethanol, isopropanol, t-butanol, diethyl ether, diphenyl ether, diisopropyl ether (DIPE), dimethylacetamide (DMA), chlorobenzene, benzene, toluene, carbon tetrachloride (CCl4), acetone, trifluoroacetic acid, chloroform (CHCl3), pyridine, and aqueous solutions thereof, or a mixed solution thereof, and preferably THF.

[0160] According to one embodiment of the present invention, the reaction may be stirred at 20 to 30°C for 3 hours.

[0161] In addition, the present invention provides a near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the above chemical formula 2.

[0162] In addition, the present invention provides a method for detecting intracellular glutathione, comprising a step of adding a nanocomplex represented by the above chemical formula 2 to a sample.

[0163] In addition, the present invention provides a method for photodynamic treatment of cancer, comprising the step of administering to a subject a pharmaceutically effective amount of a nanocomposite represented by the above chemical formula 1.

[0164] The treatment method of the present invention comprises administering the nanocomplex to a subject in a therapeutically effective amount. It is preferred that the specific therapeutically effective amount for a specific subject be applied differently depending on various factors including the type and degree of response to be achieved, the specific composition including whether other agents are used in some cases, the age, body weight, general health, sex and diet of the subject, the time of administration, the route of administration and the secretion rate of the composition, the treatment period, drugs used together or concurrently with the specific composition, and similar factors well known in the medical field. The daily dosage is 0.0001 to 100 mg / kg, preferably 0.01 to 100 mg / kg, based on the amount of the pharmaceutical composition of the present invention, and can be administered 1 to 6 times a day. However, it is obvious to those skilled in the art that the dosage or administration of each active ingredient should be such that the content of each active ingredient is not excessively high and side effects are not caused. Therefore, it is preferred that the effective amount of a composition suitable for the purpose of the present invention be determined in consideration of the aforementioned matters.

[0165] The above object is applicable to any mammal, which includes not only humans and primates, but also livestock such as cows, pigs, sheep, horses, dogs and cats.

[0166] The nanocomposite of the present invention can be administered to mammals such as rats, mice, livestock, and humans via various routes. All modes of administration are conceivable, and for example, administration can be by oral, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebroventricular injection.

[0167] Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are only intended to illustrate the present invention, and the scope of the present invention is not limited by these examples.

[0168] <Manufacturing Example 1> Preparation of nitrobenzoselenadiazole-cysteine-copper (complex 1)

[0169] To synthesize the nitrobenzoselenadiazole-cysteine-copper complex of the present invention, the following reaction scheme 1 was used.

[0170] [Reaction Formula 1]

[0171]

[0172] <1-1> Preparation of 4-fluoro-benzoselenadiazole

[0173] 50 mg (0.3964 mmol) of 3-fluorobenzene-1,2-diamine and 52.78 mg (0.4757 mmol) of selenium dioxide (SeO2) were added to 6.94 mL of ethanol, and the mixture was slowly heated under reflux at 80 °C for 1 hour. After completion of the reaction, the mixture was cooled to 25 °C, and ethyl acetate (EA) and distilled water (DI H2O) were added to separate the organic layer and the aqueous layer. The organic layer was washed three times with distilled water, dried over anhydrous sodium sulfate (Na2SO4), the solvent was filtered, and the filtrate was evaporated under reduced pressure. The above mixture was purified by flash silica gel column chromatography with a mixed solvent of EA / n-hex=1:9 by volume to obtain 45 mg (90% yield) of 4-fluoro-benzoselenadiazole as a white powder.

[0174] 1 H-NMR (500 MHz, DMSO-d6); δ 7.70 (d, J = 9.1, 7.3, 5.6 Hz, 1H), 7.31 (ddd, J = 10.8, 7.4 Hz, 1H).

[0175] 13 C-NMR (126 MHz, DMSO-d6); δ 162.18, 155.24, 153.16, 152.01(d, JC-H = 15.12 Hz), 129.88(d, JC-N = 7.56 Hz), 120.73(d, JC-N = 6.3 Hz), 111.92(d, JC-H = 16.38 Hz), 80.20(d, JC-F = 34.02, 32.76 Hz).8 MHz.9(d).

[0176] <1-2> Preparation of 4-fluoro-nitrobenzoselenadiazole

[0177] 50 mg (0.2486 mmol) of 4-fluoro-benzoselenadiazole was dissolved in 4 mL of concentrated sulfuric acid (conc. H2SO4, 0.003 mmol) at 0 °C, and 0.16 mL of concentrated nitric acid (conc. HNO3, 3.730 mmol) was added dropwise while stirring, and the mixture was stirred for 30 minutes. 10 mL of cold distilled water was added to terminate the reaction at 25 °C for 10 minutes. The yellow precipitate was filtered through Whatman Grade 1 filter paper, and the filtrate was concentrated under reduced pressure to obtain 44.5 mg (yield 89%) of 4-fluoro-nitrobenzoselenadiazole as a yellow powder.

[0178] 1 H-NMR (500 MHz, DMSO-d6) δ 8.61 (dd, J = 8.4, 4.7 Hz, 1H), 7.55 (dd, J = 9.7, 8.4 Hz, 1H).

[0179] 13 C-NMR (126 MHz, DMSO-d6); δ 157.75, 155.60, 152.40, 151.51, 138.11, 129.65 (d, JC-N = 10.08 Hz), 110.25 (d, JC-H = 18.9 Hz), 79.76 (t, JC-F = 34.02, 32.76 Hz).

[0180] <1-3> Preparation of nitrobenzoselenadiazole-cysteine-copper complex

[0181] 58 mg (0.25 mmol) of 4-fluoro-nitrobenzoselenadiazole was dissolved in 0.5 mL of tetrahydrofuran (THF), and 30.3 mg (0.25 mmol) of cysteine ​​and 2.5 mL (0.25 mmol) of aqueous sodium hydroxide solution were added. The mixture was stirred at 25 °C for 3 h, and then 2.5 mL (0.25 mmol) of aqueous CuCl2 solution was added dropwise. The reaction was maintained for 5 min to form a nanocomposite through self-assembly. The mixture was washed three times with distilled water to remove unreacted copper ions, and dried at -70 °C for 24 h to obtain 32 mg of the nitrobenzoselenadiazole-cysteine-copper complex.

[0182] <Manufacturing Example 2> Manufacturing of nitrobenzofurazan-cysteine-copper (complex 2)

[0183] To synthesize the nitrobenzofurazan-cysteine-copper complex of the present invention, the following reaction scheme 2 was used.

[0184] [Reaction Formula 2]

[0185]

[0186] <2-1> Preparation of nitrobenzofurazan-cysteine-copper complex

[0187] 49.8 mg (0.25 mmol) of 4-chloro-7-nitrobenzofurazan was dissolved in 0.5 mL of tetrahydrofuran, and 30.3 mg (0.25 mmol) of cysteine ​​and 2.5 mL (0.25 mmol) of aqueous sodium hydroxide solution were added. The mixture was stirred at 25 °C for 3 h, and then 2.5 mL (0.25 mmol) of aqueous CuCl2 solution was added dropwise. The reaction was maintained for 5 min to form a nanocomposite through self-assembly. The mixture was washed three times with distilled water to remove unreacted remaining copper ions, and dried at -70 °C for 24 h to obtain 35 mg of a nitrobenzofurazan-cysteine-copper complex.

[0188] <Manufacturing Example 3> Fluorescent probe sample manufacturing

[0189] As shown in the following reaction schemes 3 and 4, the complex 1 or complex 2 was reacted with glutathione (GSH) to prepare fluorescent probe samples of nitrobenzoselenadiazole-cysteine ​​(compound 1) and nitrobenzofurazan-cysteine ​​(compound 2).

[0190] [Reaction Formula 3]

[0191]

[0192] [Reaction Formula 4]

[0193]

[0194] Specifically, complex 1 or complex 2 stored at 4°C in methanol (MeOH) solvent was washed three times with distilled water, 6 mg of glutathione and 2 mL of distilled water were added, and the mixture was reacted at 25°C for 5 minutes. Then, centrifugation was performed at 25°C for 3 minutes using a centrifuge (Eppendorf Centrifuge model 54, Eppendorf) 3750 rcf, and 1 mL of each supernatant was added to 9 mL of distilled water to prepare compound 1 samples and compound 2 samples.

[0195] A schematic diagram of the photodynamic therapy process in cancer cells using the nanocomposite of the present invention is shown in Figure 1.

[0196] <Example 1> Analysis of the central metal coordination number of nanocomposites

[0197] In order to confirm the coordination bond number for copper ions of complex 1 and complex 2 of the present invention, Fourier transform infrared spectroscopy (Nicolet TM iS TM FT-IR spectra were measured using a 5 FT-IR spectrometer (Thermo Scientific).

[0198] As a result, as shown in Fig. 2, compared to the spectrum of cysteine, both complex 1 and complex 2 have a spectrum of 2550 to 2600 cm corresponding to the thiol group, indicating the formation of nanocomposites. -1 The peak was confirmed at the wavelength. In addition, in the case of complex 1, the electron density around the C=O bond was reduced compared to complex 2, so the peak corresponding to the C=O bond was at 1575.5 cm -1 1611.8 cm at -1 , and it can be confirmed that the C=O group of complex 1 coordinates with the divalent copper ion to show a four-coordinate structure.

[0199] <Example 2> Analysis of specific fluorescent detection substances

[0200] The selectivity was confirmed by comparing the fluorescence intensity of complex 1 or complex 2 for various biomolecules or metal ions.

[0201] Specifically, glutathione (GSH), cysteine, homocysteine, and hydrogen sulfide, citrulline, arginine, glutamine, glycine, human serum albumin, hydrogen peroxide, and ethylenediaminetetraacetic acid were used as biomolecules, and zinc chloride, copper(II) chloride, cobalt(II) chloride, cadmium chloride, sodium chloride, magnesium chloride, iron(III) chloride, potassium chloride, and mercury(II) nitrate were used as metal ion salts to form nanocomposites. The fluorescence intensity after the reaction was measured.

[0202] As a result, as shown in Fig. 3, it was confirmed that both complex 1 and complex 2 showed fluorescence when glutathione, cysteine, homocysteine, or hydrogen sulfide was added, and in particular, it was confirmed that the highest fluorescence intensity was shown in the reaction with glutathione.

[0203] <Example 3> Analysis of fluorescence emission characteristics

[0204] The presence of glutathione (GSH)-specific fluorescence detection reaction of complex 1 or complex 2 was confirmed.

[0205] Specifically, samples of complex 1 or complex 2 were applied to a standard quartz cell (1 cm, Hellma Analytics, Germany), and the fluorescence emission spectra were measured using a fluorescence measurement device (Spectrofluorophotometer; Shimadzu Corp. RF-6000, Kyo-to, Japan). In addition, a chamber with an excitation wavelength of 365 nm (WUV-L10, DAIHAN Scientific) was used to visually confirm the fluorescence change.

[0206] As a result, as shown in Fig. 4, it was confirmed that the compound produced by the reaction of complex 1 and glutathione showed an increase in fluorescence intensity at a wavelength of 600 nm to 650 nm, and it was confirmed that the compound produced by the reaction of complex 2 and glutathione showed an increase in fluorescence intensity at a wavelength of 520 nm to 580 nm.

[0207] <Example 4> Confirmation of reactive oxygen species

[0208] <4-1> Confirmation of generation of reactive oxygen species by hydrogen peroxide and light irradiation

[0209] The generation of reactive oxygen species through the reaction of Cu(I) ions generated by the reaction of complex 1 or complex 2 with glutathione and hydrogen peroxide, and the generation of reactive oxygen species through light irradiation of compound 1 or compound 2 from which copper was removed after the glutathione reaction, were confirmed using 1,3-diphenylisobenzofuran, a standard reagent for measuring reactive oxygen species.

[0210] Specifically, complex 1 or complex 2 (4 mg / mL) was washed three times with distilled water, and each supernatant was diluted 100-fold by adding glutathione (12.5 mM). Then, the diluted solution and 1,3-diphenylisobenzofuran (100 μM) dimethyl sulfoxide (DMSO) solution were mixed in distilled water to prepare samples, and each sample was irradiated with light (532 nm, 45 mW cm) for 60 seconds. -2) and the absorption spectrum of 1,3-diphenylisobenzofuran was measured for up to 360 seconds.

[0211] As a result, as shown in Fig. 5, it was confirmed that the sample to which glutathione was added to complex 1 had a decrease in absorbance at a wavelength of 400 to 500 nm due to an increase in reactive oxygen species through hydrogen peroxide or light irradiation, and it was confirmed that the sample to which glutathione was added to complex 2 had a decrease in absorbance at a wavelength of 400 to 700 nm due to an increase in reactive oxygen species through hydrogen peroxide.

[0212] <4-2> Detection of reactive oxygen species

[0213] Detection of reactive oxygen species for complex 1 or complex 2 was confirmed using an electron paramagnetic resonance (EMXplus-9.5 / 12 / P / L System, Germany) analyzer.

[0214] Specifically, glutathione aqueous solution (50 mM) was added to complex 1 (4 mg / mL) or complex 2 (4 mg / mL), stirred for 5 minutes, and the supernatant of the mixture was obtained, and singlet oxygen ( 1 O2) 2,2,6,6-Tetramethylpiperidine (TEMP) as a detector and 5,5-Dimethyl-1-pyrroline N-oxide (DMPO) as a reactive oxygen species detector were added to the glutathione reaction product sample, the glutathione reaction and hydrogen peroxide addition sample, and the glutathione reaction and light irradiation sample, respectively, and then the amount of singlet oxygen production was measured for each sample.

[0215] As a result, as shown in Fig. 6, it was confirmed that the samples in which glutathione was added to complex 1 or complex 2 showed an increase in the amount of singlet oxygen and reactive oxygen species produced under hydrogen peroxide, and it was confirmed that only compound 1 generated singlet oxygen and hydroxyl radicals by light irradiation.

[0216] <Example 5> Intracellular fluorescence imaging

[0217] <5-1> Detection of intracellular glutathione fluorescence

[0218] Glutathione-specific detection and fluorescence imaging of complex 1 or complex 2 within tumor cells were confirmed.

[0219] Specifically, experimental group 1, in which the U87 MG (human brain tumor cell line) cell line was pretreated with N-ethylmaleimide (NEM) (50 μM), a glutathione inhibitor, for 30 minutes to reduce the intracellular glutathione level, experimental group 2, in which glutathione (500 μM) was additionally treated to the above experimental group 1 to enrich the intracellular glutathione, and the control group to which no glutathione inhibitor was added, were pretreated and incubated with complex 1 or complex 2 at a concentration of 100 μg / mL for 1 hour, and then intracellular fluorescence was measured using a confocal laser scanning microscope (LSM800, Carl Zeiss, Germany).

[0220] As a result, as shown in Fig. 7, when complex 1 or complex 2 was treated in the glutathione-suppressed experimental group 1, the fluorescence emission intensity decreased, and when complex 1 or complex 2 was treated in the glutathione-rich experimental group 2 and the tumor cell control group, the fluorescence emission intensity increased.

[0221] <5-2> Detection of intracellular reactive oxygen species

[0222] The ability of complex 1 or complex 2 to generate reactive oxygen species through intracellular glutathione-specific reactions was confirmed.

[0223] Specifically, human kidney cell HEK293 cell line and tumor cell U87 MG cell line were treated with a solution of 2',7'-dichlorofluorescein diacetate (DCFDA), a fluorescent probe, and treated with complex 1 or complex 2 at a concentration of 100 μg / mL, respectively, and incubated for 90 minutes, followed by 1 minute of light irradiation (532 nm, 45 mW cm -2 ) and monitored the reactive oxygen species generated.

[0224] As a result, as shown in Fig. 8, the normal cell group did not exhibit fluorescence, whereas the U87 MG tumor cell group treated with complex 1 exhibited green fluorescence, and it was confirmed that the fluorescence intensity increased significantly, particularly in the group treated with light irradiation.

[0225] Therefore, it can be confirmed that reactive oxygen species are significantly generated in the tumor cell group treated with complex 1 due to the high concentration of glutathione in cancer cells compared to normal cells.

[0226] <Example 6> In vivo anticancer efficacy

[0227] <6-1> Tumor size analysis

[0228] The cancer-specific therapeutic efficacy of complex 1 was confirmed in a xenograft mouse model.

[0229] Specifically, to induce tumors in mouse models, U87 MG cell lines were inserted into the right thigh of BALB / c nude mice, and complex 1 (10 mg / kg) alone or complex 1 (10 mg / kg) was administered subcutaneously to the tumor site and treated with light irradiation (532 nm, 45 mW cm -2, 1 min) and a 4-day rest period were included, and the administration cycle was repeated 5 times for 20 days, and weight changes and other symptoms were monitored.

[0230] As a result, as shown in Fig. 9, the tumor size in the group administered complex 1 alone or the group treated with complex 1 and light irradiation was 70.59 mm, respectively. 3 62.51 mm in 3 , 63.96 mm 3 42.03 mm in 3 It was confirmed that the weight change was maintained constant.

[0231] <6-2> Analysis of tumor antiangiogenesis-related proteins

[0232] After sacrificing the U87 MG transplanted mouse model on day 20, the tumors were surgically removed and factors contributing to tumor size reduction were analyzed.

[0233] Specifically, for the tumors of the PBS (control group), complex 1 administration group, complex 1 administration and light irradiation treatment group, the quantification of tumor angiogenesis-related proteins was performed using the Pierce BCA Protein Assay Kit (Thermo Scientific, USA), and for the tumor tissues, the Proteome Profiler Mouse Angiogenesis Array Kit (R&D Systems, Inc., USA) was used, and it was performed based on the sandwich-antigen-antibody methodology. In the final step, the antigen-antibody mixture was applied to the antibody-coated panel, and then analyzed using an enhanced chemiluminescence (ECL) solution.

[0234] Complex 1 single administration groupComplex 1 administration and light irradiation treatment groupIncreased protein expression-Endostatin / collagen XVIII, MMP-3, Coagulation Factor III, IL-1a, PEDF, IL-1b, CD26-Decreased protein expression-Endostatin / collagen XVIII, MMP-3, Coagulation Factor III, IL-1a, PEDF, IL-1b, CD26

[0235] As a result, as shown in Fig. 10 and Table 1 above, protein expression heat-mapping analysis showed that, compared to the PBS control group, the cell group administered complex 1 alone showed increased expression of Endostatin / collagen XVIII, MMP-3, Coagulation Factor III, IL-1a, PEDF, IL-1b, and CD26 proteins, indicating potential promotion of angiogenesis, whereas the group administered complex 1 and treated with light showed a decrease in the protein levels of the angiogenic factors. Therefore, it can be confirmed that intracellular complex 1 exhibits an antiangiogenic effect by decreasing the expression of angiogenesis-related proteins due to the generation of reactive oxygen species by light irradiation.

[0236] Therefore, the nanocomposite NPS-Cys-Cu of the present invention 2+ It specifically reacts with glutathione in cancer cells, enabling near-infrared imaging, and Cu reduced by reaction with glutathione + The effect of increasing the expression of angiogenesis-related proteins by generating reactive oxygen species through reaction with hydrogen peroxide was confirmed. In addition, the compound NPS-Cys formed by the complex and glutathione reaction was irradiated with light to generate reactive oxygen species, thereby reducing the expression of angiogenesis-related proteins, confirming its anticancer effect.

[0237] As described above, specific embodiments of the present invention have been described in detail. However, those skilled in the art who understand the spirit of the present invention will be able to easily suggest other inventions that are backwards or other embodiments included within the scope of the spirit of the present invention by adding, changing, or deleting other components within the scope of the same spirit. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention is indicated by the scope of the claims described below rather than the detailed description described above, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A nanocomposite represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] (In the above chemical formula 1, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy, M is a divalent cation metal.) 2. In paragraph 1, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the metal comprises copper.

3. In paragraph 1, The nanocomposite is a nanocomposite or a pharmaceutically acceptable salt thereof, which is a complex as follows.

4. In paragraph 1, The nanocomplex is a nanocomplex that specifically reacts with glutathione, or a pharmaceutically acceptable salt thereof.

5. In paragraph 1, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the nanocomposite reacts with glutathione to form a compound represented by the following chemical formula 3. [Chemical Formula 3] (In the above chemical formula 3, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, or alkoxy.) 6. In paragraph 5, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the compound is the following compound.

7. In paragraph 5, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the compound emits fluorescence at 400 to 700 nm.

8. In paragraph 1, The nanocomposite or a pharmaceutically acceptable salt thereof, wherein the nanocomposite reacts with glutathione to reduce a divalent cation metal to a monovalent cation metal.

9. In paragraph 8, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the above monovalent cationic metal reacts with a biomolecule to generate reactive oxygen species.

10. In paragraph 9, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the biomolecule comprises hydrogen peroxide, an amino acid, a protein, a lipid, a by-product of cell metabolism, or an active substance comprising oxygen.

11. In paragraph 9, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the reactive oxygen species comprises a hydroxyl radical, singlet oxygen, hydrogen peroxide, superoxide, or nitric oxide.

12. In paragraph 1, The nanocomplex or a pharmaceutically acceptable salt thereof, wherein the nanocomplex increases the expression of Endostatin / collagen XVIII, MMP-3, Coagulation Facter III, IL-1a, PEDF, IL-1b or CD26 proteins.

13. In paragraph 5, The compound is a nanocomposite or a pharmaceutically acceptable salt thereof, which reduces the expression of Endostatin / colla gen XVIII, MMP-3, Coagulation Facter III, IL-1a, PEDF, IL-1b or CD26 proteins upon photoirradiation.

14. A pharmaceutical composition for photodynamic therapy of cancer, comprising a nanocomposite represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical Formula 1] (In the above chemical formula 1, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently one of hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, and alkoxy, M is a divalent cation metal.) 15. In paragraph 14, A composition wherein the cancer is at least one selected from the group consisting of brain cancer, spinal cancer, lung cancer, thyroid cancer, breast cancer, skin cancer, prostate cancer, and ovarian cancer.

16. In paragraph 14, The composition above is a composition that inhibits angiogenesis of cancer.

17. A method for producing a nanocomposite manufactured by the following reaction scheme 1, [Reaction Formula 1] A step of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent; and A method for producing a nanocomposite, comprising the step of adding copper cations to the above reactants.

18. In paragraph 17, The above organic solvents are tetrahydrofuran (THF), dimethylformamide (DMF), and dichloromethane (CH 2 Cl 2 ), 1,2-dichloroethane (CH 2 ClCH 2 Cl), dimethyl sulfoxide (DMSO), t-butyl methyl ether (TBME), acetonitrile (ACN), methanol, ethanol, isopropanol, t-butanol, diethyl ether, diphenyl ether, diisopropyl ether (DIPE), dimethylacetamide (DMA), chlorobenzene, benzene, toluene, carbon tetrachloride (CCl 4 ), acetone, trifluoroacetic acid, chloroform (CHCl 3 ), pyridine and their aqueous solutions, or a mixed solution thereof.

19. In Article 17, A method wherein the above reaction is stirred at 20 to 30°C for 3 hours.

20. A near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the following chemical formula 1. [Chemical Formula 1] (In the above chemical formula 1, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently one of hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, and alkoxy, M is a divalent cation metal.) 21. A method for detecting intracellular glutathione, comprising the step of adding a nanocomposite represented by the following chemical formula 1 to a sample. [Chemical Formula 1] (In the above chemical formula 1, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently one of hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, and alkoxy, M is a divalent cation metal.) 22. A nanocomposite represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof: [Chemical formula 2] (In the above chemical formula 2, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy, M is a divalent cation metal.) 23. In paragraph 22, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the metal is copper.

24. In paragraph 22, The nanocomposite is a nanocomposite or a pharmaceutically acceptable salt thereof, which is a complex as follows.

25. In paragraph 22, The nanocomplex is a nanocomplex that specifically reacts with glutathione, or a pharmaceutically acceptable salt thereof.

26. In paragraph 22, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the nanocomposite reacts with glutathione to form a compound represented by the following chemical formula 3. [Chemical Formula 3] (In the above chemical formula 3, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl, or alkoxy.) 27. In paragraph 26, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the compound is the following compound.

28. In paragraph 26, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the compound emits fluorescence at 400 to 700 nm.

29. In paragraph 22, The nanocomposite or a pharmaceutically acceptable salt thereof, wherein the nanocomposite reacts with glutathione to reduce a divalent cation metal to a monovalent cation metal.

30. In paragraph 29, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the above monovalent cationic metal reacts with a biomolecule to generate reactive oxygen species.

31. In paragraph 30, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the biomolecule comprises an active substance including hydrogen peroxide, an amino acid, a protein, a lipid, a by-product of cell metabolism, or oxygen.

32. In paragraph 30, A nanocomposite or a pharmaceutically acceptable salt thereof, wherein the reactive oxygen species comprises a hydroxyl radical, singlet oxygen, hydrogen peroxide, superoxide, or nitric oxide.

33. A method for producing a nanocomposite manufactured by the following reaction formula 2, [Reaction Formula 2] A step of reacting 4-fluoro-nitrobenzoselenadiazole and cysteine ​​in an organic solvent; and A method for producing a nanocomposite, comprising the step of adding copper cations to the above reactants.

34. In paragraph 33, The above organic solvents are tetrahydrofuran (THF), dimethylformamide (DMF), and dichloromethane (CH 2 Cl 2 ), 1,2-dichloroethane (CH 2 ClCH 2 Cl), dimethyl sulfoxide (DMSO), t-butyl methyl ether (TBME), acetonitrile (ACN), methanol, ethanol, isopropanol, t-butanol, diethyl ether, diphenyl ether, diisopropyl ether (DIPE), dimethylacetamide (DMA), chlorobenzene, benzene, toluene, carbon tetrachloride (CCl 4 ), acetone, trifluoroacetic acid, chloroform (CHCl 3 ), pyridine and their aqueous solutions, or a mixed solution thereof.

35. In paragraph 33, A method wherein the above reaction is stirred at 20 to 30°C for 3 hours.

36. A near-infrared fluorescence sensor for detecting glutathione comprising a nanocomposite represented by the following chemical formula 2. [Chemical formula 2] (In the above chemical formula 2, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy, M is a divalent cation metal.) 37. A method for detecting intracellular glutathione, comprising the step of adding a nanocomposite represented by the following chemical formula 2 to a sample. [Chemical formula 2] (In the above chemical formula 2, A is oxygen (O) or selenium (Se), X 1 Inland X 4 are each independently hydrogen, alkyl, hydroxy, thiol, amine, carbonyl, carboxy, alkenyl, alkynyl or alkoxy, M is a divalent cation metal.) 38. A method for photodynamic treatment of cancer, comprising the step of administering to a subject a pharmaceutically effective amount of a nanocomposite represented by the following chemical formula 1. [Chemical Formula 1]

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