Pharmaceutical composition comprising novel terephthalate-cyclohexylamine-based polymer compound for photodynamic therapy for glioblastoma

Abstract

The present invention relates to a pharmaceutical composition comprising a novel terephthalate-cyclohexylamine-based polymer compound for photodynamic therapy for glioblastoma, wherein the polymer compound can specifically fluorescently label glioblastoma, and generates reactive oxygen species in glioblastoma so as to reduce the size of tumor cells such that an anticancer effect is exhibited, and thus the present invention can be effectively used in related industries.

Description

Pharmaceutical composition for photodynamic therapy of glioblastoma comprising novel polymeric compounds based on terephthalate-cyclohexylamine

[0001] The present invention relates to a pharmaceutical composition for photodynamic therapy of glioblastoma comprising a novel polymer compound based on terephthalate-cyclohexylamine.

[0002] Glioblastoma is a tumor that arises from glial cells throughout the brain. It accounts for approximately 12-15% of all brain tumors, making it a fatal brain tumor in humans. The exact cause of glioblastoma is unknown, but it is believed to be related to genetic and environmental factors. Furthermore, glioblastoma is a primary malignant brain tumor that occurs in adults. If left untreated, it can lead to death within 3-6 months after diagnosis. Even with treatment, the average survival time is only 12-14 months, making it a highly malignant tumor. Treatment methods for glioblastoma include direct surgery, as well as adjuvant treatments such as chemotherapy and radiation therapy. Only recently has photodynamic therapy gained recognition as an adjuvant treatment for the tumor.

[0003] Meanwhile, 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 has the advantage of allowing for precise treatment of specific areas. Therefore, PDT can enhance the production of ROS and can complement chemodynamic therapy. Only a handful of FDA-approved photosensitizers for PDT have been used, and photosensitizers with various wavelengths are continuously being developed. In particular, single-benzene-based fluorescent probes are being studied in various fields, and some of these fluorescent probes can be utilized as photosensitizers.

[0004] Accordingly, the inventors of the present invention have developed a novel photosensitizer based on a single-benzene polymer that can be manufactured by a simple synthetic method and can form a large amount of reactive oxygen species in a short period of time, and have confirmed that it can be used as an adjuvant treatment for photodynamic therapy in glioblastoma, thereby completing the present invention.

[0005] This patent was supported by the Ministry of Science and ICT (National Research Foundation of Korea) (Project No. 1711192122, Subproject No. 2022R1F1A1069954).

[0006] This patent was supported by the Ministry of Education (National Research Foundation of Korea) (Project No.: 1345365662, Subproject No.: 2018R1A6A1A03025124).

[0007] This patent was supported by the Ministry of Science and ICT (Science and ICT Commercialization Promotion Agency) (RS-2024-00418924).

[0008] The purpose of the present invention is to provide a polymer compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0009] [Chemical Formula 1]

[0010]

[0011] Another object of the present invention is to provide a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0012] [Chemical Formula 2]

[0013]

[0014] Another object of the present invention is to provide a method for producing a polymer compound prepared by the following reaction scheme 1, which comprises the steps of mixing dimethyl 1,4-cyclohexanedione-2,5-dicarboxylate and trans-1,4-cyclohexanediamine in an organic solvent; and adding an acid catalyst to the mixture.

[0015] [Reaction Formula 1]

[0016]

[0017] Another object of the present invention is to provide a pharmaceutical composition for photodynamic treatment of glioblastoma, comprising a polymer compound 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 photodynamic treatment of glioblastoma, comprising the step of administering to a subject a pharmaceutically effective amount of a polymer compound represented by the above chemical formula 1.

[0019] To achieve the above purpose,

[0020] The present invention provides a polymer compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0021] [Chemical Formula 1]

[0022]

[0023] In addition, the present invention provides a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0024] [Chemical Formula 2]

[0025]

[0026] In addition, the present invention provides a method for producing a polymer compound prepared by the following reaction scheme 1, including the steps of mixing dimethyl 1,4-cyclohexanedione-2,5-dicarboxylate and trans-1,4-cyclohexanediamine in an organic solvent; and adding an acid catalyst to the mixture.

[0027] [Reaction Formula 1]

[0028]

[0029] In addition, the present invention provides a pharmaceutical composition for photodynamic treatment of glioblastoma, comprising a polymer compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0030] In addition, the present invention provides a method for photodynamic treatment of glioblastoma, comprising the step of administering to a subject a pharmaceutically effective amount of a polymer compound represented by the above chemical formula 1.

[0031] The novel polymer compound of the present invention can specifically fluorescently label glioblastoma and has been confirmed to exhibit an anticancer effect by generating reactive oxygen species within glioblastoma and reducing the size of tumor cells, and thus can be usefully utilized in related industries.

[0032] Figure 1 shows the particle size, surface charge, molecular weight, FT-IR spectrum, and SEM image of a polymer compound according to the present invention.

[0033] Figure 2 shows the fluorescence stability of the polymer compound according to the present invention with respect to time, pH, and solvent.

[0034] Figure 3 shows the formation of active oxygen species in a polymer compound according to the present invention.

[0035] Figure 4 shows the intracellular toxicity and fluorescence imaging of a polymer compound according to the present invention.

[0036] Figure 5 illustrates a density functional theory regarding the generation of reactive oxygen species by a polymer compound according to the present invention.

[0037] Figure 6 shows the in vivo stability of the polymer compound according to the present invention.

[0038] Figure 7 shows the photodynamic therapy of a glioblastoma mouse model using a polymer compound according to the present invention.

[0039] 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.

[0040] 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.

[0041] Hereinafter, terms used in the present invention are explained.

[0042] The present invention provides a polymer compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0043] [Chemical Formula 1]

[0044]

[0045] The compound represented by the chemical formula 1 of the present invention can be named Polymeric amino-single-benzene (PA).

[0046] In the above chemical formula 1, n may be an integer from 1 to 100, preferably an integer from 1 to 50, and more preferably an integer from 1 to 12.

[0047] 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.

[0048] The "polymer" of the present invention is a molecule having a molecular weight of 10,000 or more, which is formed by a chemical reaction of monomer molecules and consists of a long chain with regular repeating units.

[0049] According to one embodiment of the present invention, the compound may have a particle size of 390 to 450 nm.

[0050] According to one embodiment of the present invention, the compound may emit fluorescence at 600 to 610 nm.

[0051] According to one embodiment of the present invention, the compound may have a surface charge of 1.5 to 4.8 mV.

[0052] According to one embodiment of the present invention, the compound may have a number average molecular weight and a weight average molecular weight of 3,770 and 3,881, respectively.

[0053] In addition, the present invention provides a compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof.

[0054] [Chemical Formula 2]

[0055]

[0056] The compound represented by the above chemical formula 2 of the present invention can be named Polymeric amino-single-benzene nano-aggregates (PANA).

[0057] The compound represented by the above chemical formula 2 of the present invention can be a monomer of the polymer compound represented by the above chemical formula 1 and can be used as a fluorescent substance.

[0058] The "monomer" or "unit" of the present invention is a molecule that can react with other monomer molecules to form larger polymer chains or three-dimensional networks through a process called polymerization.

[0059] 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.

[0060] In addition, the present invention provides a method for producing a polymer compound prepared by the following reaction scheme 1, including the steps of mixing dimethyl 1,4-cyclohexanedione-2,5-dicarboxylate and trans-1,4-cyclohexanediamine in an organic solvent; and adding an acid catalyst to the mixture.

[0061] [Reaction Formula 1]

[0062]

[0063] According to one embodiment of the present invention, the organic solvent may be any one selected from the group consisting of, but not limited to, ethanol, methanol, isopropanol, t-butanol, tetrahydrofuran (THF), dimethylformamide (DMF), dichloromethane (CH2Cl2), 1,2-dichloroethane (CH2ClCH2Cl), dimethyl sulfoxide (DMSO), t-butyl methyl ether (TBME), acetonitrile (ACN), 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 may be ethanol, methanol, isopropanol, or t-butanol, and more preferably may be ethanol.

[0064] According to one embodiment of the present invention, the method may further include a step of dispersing the polymer compound in an aqueous solution and performing ultrasonic treatment.

[0065] According to one embodiment of the present invention, the acid catalyst may include, but is not limited to, acetic acid, nitric acid, hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, perchloric acid, hydrofluoric acid or aluminum chloride, and preferably may be acetic acid.

[0066] According to one embodiment of the present invention, the mixture may be heated and stirred at 80° C. for 16 hours.

[0067] According to one embodiment of the present invention, the ultrasonic treatment may be performed at 200 W for 10 minutes.

[0068] According to one embodiment of the present invention, the ultrasonic treatment may be to nanoparticleize the polymer compound.

[0069] In addition, the present invention provides a pharmaceutical composition for photodynamic treatment of glioblastoma, comprising a polymer compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0070] 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.

[0071] 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.

[0072] 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.

[0073] 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.

[0074] 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.

[0075] 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.

[0076] The "photodynamic therapy (PDT)" of the present invention is a cancer treatment method that selectively destroys cancer cells by irradiating them with a laser after a certain period of time has passed after administering the photosensitizing substance, utilizing the property of photosensitizing substances accumulating in cancer cells for a long time.

[0077] According to one embodiment of the present invention, the compound may generate reactive oxygen species by light irradiation.

[0078] The "reactive oxygen species (ROS)" or "active oxygen species" of the present invention are chemically reactive molecules containing oxygen atoms. These compounds are produced within living organisms and are highly oxidizing oxygen species that attack biological tissues, damage cells, and contribute to obesity. Reactive oxygen species are produced during normal cellular activity and are involved in various biological processes, including cell differentiation, gene expression, and cytokine responses.

[0079] 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 singlet oxygen, superoxide, hydrogen peroxide, amino acids, proteins, lipids, by-products of cell metabolism, and oxygen-containing reactive substances, and preferably may be singlet oxygen or superoxide.

[0080] According to one embodiment of the present invention, the compound may specifically fluorescently label glioblastoma.

[0081] According to one embodiment of the present invention, the compound may penetrate the blood-brain barrier by 0.1 to 20 μm.

[0082] According to one embodiment of the present invention, the compound may reduce tumor size.

[0083] In addition, the present invention provides a method for photodynamic treatment of glioblastoma, comprising the step of administering to a subject a pharmaceutically effective amount of a polymer compound represented by the above chemical formula 1.

[0084] The treatment method of the present invention comprises administering to a subject a therapeutically effective amount of the polymer compound. 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 the 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 simultaneously 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 does not cause side effects. 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.

[0085] 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.

[0086] The polymer compound 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.

[0087] 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.

[0088] <Example 1> Preparation of a terephthalate-cyclohexylamine-based polymer compound

[0089] In order to synthesize a polymer compound represented by the following chemical formula 1, which includes a compound represented by the following chemical formula 2 of the present invention as a monomer, the following reaction scheme 1 was used.

[0090] [Chemical Formula 1]

[0091]

[0092] [Chemical Formula 2]

[0093]

[0094] [Reaction Formula 1]

[0095]

[0096] Specifically, 10 g of dimethyl 1,4-cyclohexanedione-2,5-dicarboxylate (DCD) and 7.5 g of trans-1,4-cyclohexanediamine were added to 50 mL of ethanol, stirred at 250 rpm and 80 °C for 30 minutes, and then 5 mL of acetic acid was additionally added and stirred at 80 °C for 16 hours. After confirming the completion of the reaction, 6 mL of water was added to terminate the reaction, and the mixture was washed with ethyl acetate (EtOAc), filtered, and concentrated to obtain 17 g of the product. The product was dispersed in distilled water and sonicated at 200 W for 10 minutes to produce 17 g of a nano-sized polymer.

[0097] 1 H-NMR (800 MHz, DMSO-D6): δ 9.86, 7.75, 7.27, 7.23, 6.70, 6.69, 6.61, 6.60, 6.51, 5.60, 5.39, 4.34, 4.33, 3.84, 3.82, 3.45, 3.44, 3.43, 3.42, 3.31, 3.29, 2.50, 2.49, 1.90, 1.06, 1.05, 1.04.

[0098] <Example 2> Analysis of structural properties of polymer compounds

[0099] <2-1> Particle size analysis of polymer compounds

[0100] In order to confirm the particle size of the polymer compound of the present invention, the particle size was measured over time under distilled water solvent conditions using a dynamic light scattering device (Malvern Instruments Zetasizer Nano ZS90), and TEM images were taken using a transmission electron microscope device (Tecnai G2 F30ST).

[0101] As a result, as shown in Figures 1a and 1e, the particle size of the polymer compound was found to be 396 to 444 nm, which was similar to the average size of 400 nm of actual nanoparticles measured in TEM images.

[0102] <2-2> Surface charge analysis of polymer compounds

[0103] The surface charge of polymer compounds over time was measured in distilled water solvent conditions using dynamic light scattering equipment.

[0104] As a result, as shown in Fig. 1b, it was confirmed that the initial surface charge of the polymer compound was 4.76 mV, 3.10 mV after 30 minutes, and 1.58 mV after 60 minutes.

[0105] <2-3> Structural analysis of polymer compounds

[0106] In order to confirm whether the polymer compound of the present invention was synthesized, the molecular weight was measured under tetrahydrofuran solvent conditions using gel permeation chromatography (GPC), and Thermo Scientific Nicolet TM iS TM FT-IR spectra of polymer compounds were measured using 5 (Thermo Fisher Scientific, USA).

[0107] As a result, as shown in Figures 1c and 1d, the number average molecular weight and weight average molecular weight (Mw) of the polymer compound were 3,770 and 3,881, and in the FT-IR spectrum of the polymer compound, the peak corresponding to the CH of the methyl group of DCD was 2800-3000 cm -1 , the peak corresponding to the NH group of cyclohexanediamine is 3200-3400 cm -1 By confirming that it appeared in , it was confirmed that a polymer compound was synthesized.

[0108] <2-4> Component analysis of polymer compounds

[0109] To confirm the elemental content of the polymer compound, the elemental components were qualitatively analyzed using EDS (Energy-dispersive X-ray spectroscopy) equipment (Tecnai G2 F30ST).

[0110] As a result, as shown in Fig. 1e, it was confirmed that the carbon, nitrogen, and oxygen of the polymer compound were 96.04%, 1.36%, and 1.41%, respectively.

[0111] <Example 3> Analysis of optical properties of polymer compounds

[0112] In order to confirm the absorption and fluorescence emission characteristics of the polymer compound of the present invention, the absorption and fluorescence spectra were measured at an excitation wavelength of 514 nm using a UV / Vis spectrophotometer (Agilent Technologies Cary 8454, USA) and a fluorescence spectrophotometer (SHIMADZU CORP. RF-6000, Japan) at a concentration of 10 μM in distilled water, ethanol, acetonitrile, DMSO, tetrahydrofuran or DMF, respectively, in a standard quartz cell (standard quartz cell, internal volume = 0.1 cm) with a thickness of 1 cm.

[0113] As a result, as shown in Fig. 2, the absorption spectrum showed a maximum absorption wavelength at 514 nm, and the fluorescence emission spectrum confirmed that the fluorescence intensity increased at 604 nm.

[0114] In addition, the polymer compound sample maintained a constant fluorescence intensity for 60 minutes, the fluorescence intensity of the polymer compound was constant at pH 3 to 9, and the polymer compound sample showed fluorescence using various solvents, confirming fluorescence stability.

[0115] <Example 4> Analysis of reactive oxygen species production in polymer compounds

[0116] To identify reactive oxygen species generated by photoirradiation of polymer compounds, 10 μM of the polymer compound and 10 μM of 9,10-diphenylanthracene (DPA), a probe of reactive oxygen species, were added to distilled water, and the absorbance was measured by irradiating with light sources of excitation wavelengths of 405 and 530 nm. In addition, to identify the type of reactive oxygen species, 10 μM of 2,2,6,6-tetramethylpiperidine (TEMP) was additionally added, and electron paramagnetic resonance (EPR) was measured using the electron paramagnetic resonance (EPR) experimental method.

[0117] As a result, as shown in Fig. 3, singlet oxygen ( 1 O2) and superoxide radicals (O2 ·- ) was generated, and when irradiated with a 405 nm light source, it was confirmed that reactive oxygen species were formed for 40 seconds, and when irradiated with a 530 nm light source, it was confirmed that reactive oxygen species were formed for 25 minutes.

[0118] <Example 5> Analysis of intracellular polymer compound activity

[0119] <4-1> Cytotoxicity analysis

[0120] Cytotoxicity according to the presence or absence of light irradiation was determined by checking the number of cells through light absorption using Cell Counting Kit-8. In order to confirm the cytotoxicity of the polymer compound of the present invention, cytotoxicity was evaluated by monitoring reactive oxygen species generated in HEK293 and b.End3 normal cells or U87 MG (human brain tumor cell line) tumor cell line.

[0121] As a result, as shown in Fig. 4a, the HEK293 and b.End3 normal cell groups did not show toxicity to the polymer compound, while the U87 MG tumor cell group showed toxicity to the polymer compound, and in particular, it was confirmed that the tumor cell viability was significantly reduced to 20% in the light-irradiated group.

[0122] <4-2> Fluorescence imaging and cell permeability analysis

[0123] In order to confirm the permeability of the polymer compound of the present invention into U87 MG tumor cells, 3000 b.End 3 cells were cultured in the upper chamber of a transwell for 7 days to artificially form a blood barrier in vitro, and a blood-brain barrier model manufactured at the cellular level was manufactured, and then treated with a polymer compound at a concentration of 1 M.

[0124] As a result, as shown in Figures 4b to 4e, the blood-brain barrier permeability of the polymer compound was 80% for 12 hours, and it was confirmed that the fluorescence intensity within U87 MG tumor cells increased for 24 hours and that it penetrated into the cytoplasm within the cells.

[0125] <Example 6> Density functional theory

[0126] In order to confirm the efficiency of generating reactive oxygen species according to the number of monomer polymerizations of the polymer compound of the present invention, quantum chemical calculations were performed using density functional theory (DFT) for monomers, dimers, and trimers of the compound, and in order to confirm the reaction rate of intersystem crossing (ISC) by light irradiation, the ISC reaction rate constant was calculated using the following mathematical equation 1.

[0127] [Mathematical Formula 1]

[0128] [Correction pursuant to Rule 91, December 24, 2024]

[0129] As a result, as shown in Fig. 5, the HOMO and LUMO energy levels of the single molecule, dimer, and trimer of the compound were -4.74 to -4.79 eV and -1.78 to -1.80 eV, respectively, and the energy band gap was all 2.96 eV.

[0130] In addition, the values ​​of the intersystem crossing constants of the compound's monomer, dimer, and trimer are 6.8×10 4 s -1 , 1.3×10 2 s -1 and 1.6×10 2 s -1 It was calculated as , and it was confirmed that the reaction speed of the trimer increased by 105 times compared to that of the single molecule.

[0131] Therefore, it can be confirmed that as the polymerization of the compound of the present invention increases, the efficiency of generating active oxygen species increases.

[0132] <Example 7> Safety analysis of polymer compounds in mice

[0133] In order to confirm the in vivo safety of the polymer compound of the present invention, blood was collected from a mouse (DBL Co., Ltd.), and red blood cells were separated by centrifugation. Then, 0.1 to 1 mg of the polymer compound was treated in 1 mL of a solvent containing 8% red blood cells, and an in vitro hemolytic reaction was confirmed through absorbance analysis. In addition, 0.3 mg / mL of the polymer compound was intravenously administered to mice for 7 days, and changes in body weight and various immune factors were analyzed to conduct an in vivo short-term toxicity evaluation.

[0134] As a result, as shown in Fig. 6, no in vitro hemolytic reaction was observed at 0.1 to 1 mg / mL of the polymer compound, and no changes in the numbers of immune factors B cells, T cells (CD3+), CD4+, and CD8+, changes in body weight, lesions in organs, changes in the numbers of AST, ALT, and BUN in the liver and kidney, and visual changes were observed for 7 days after treatment with 0.3 mg / mL of the polymer compound in the body.

[0135] <Example 8> Analysis of the efficacy of photodynamic therapy in glioblastoma mice

[0136] In order to confirm the photodynamic therapeutic efficacy of the polymer compound of the present invention on glioblastoma, 5.0 Х 10 LUC-U87MG glioblastoma cells were cultured in 5 μL of cell culture media from which fetal bovine serum (FBS) was removed. 5 5 μL of the solution containing the dog was administered to the thalamus of the mouse brain, and a glioblastoma mouse model was formed 14 days later. After intravenous administration of 0.3 mg / mL of the polymer compound at two-day intervals for 6 days and irradiation with a light source, 150 mg / kg (n=10) of luciferin (D-Luciferin) was administered intraperitoneally to the mice, and the brain of the glioblastoma mouse model was imaged using a fluorescence tissue imaging system (FTIS; Fluorescence tissue imaging system; VISQUE InVivo Elite. Vieworks) and the fluorescence emission signal (intensity) was measured.

[0137] As a result, as shown in Fig. 7, the tumor size of the experimental group treated with the polymer compound and light irradiation was significantly reduced compared to the control group over 28 days, and body weight was confirmed to remain constant. Furthermore, the survival rate of the experimental group treated with the polymer compound and light irradiation increased, and imaging confirmed that the polymer compound accumulated in the brain, lungs, liver, and kidneys of the mouse model.

[0138] Therefore, it was confirmed that the terephthalate-cyclohexylamine-based polymer compound of the present invention can specifically fluorescently label glioblastoma and exhibit an anticancer effect by generating reactive oxygen species within glioblastoma and reducing the size of tumor cells.

[0139] 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 polymer compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] 2. In paragraph 1, The compound is a polymer compound or a pharmaceutically acceptable salt thereof, having a particle size of 390 to 450 nm.

3. In paragraph 1, The compound is a polymer compound or a pharmaceutically acceptable salt thereof, which emits fluorescence at 600 to 610 nm.

4. In paragraph 1, The compound is a polymer compound or a pharmaceutically acceptable salt thereof, wherein the surface charge is 1.5 to 4.8 mV.

5. In paragraph 1, The compound is a polymer compound or a pharmaceutically acceptable salt thereof, wherein the number average molecular weight and the weight average molecular weight are 3,770 and 3,881, respectively.

6. A compound represented by the following chemical formula 2 or a pharmaceutically acceptable salt thereof: [Chemical formula 2] 7. A method for producing a polymer compound represented by the following chemical formula 1, which is produced by the following reaction formula 1, [Chemical Formula 1] [Reaction Formula 1] A step of mixing dimethyl 1,4-cyclohexanedione-2,5-dicarboxylate and trans-1,4-cyclohexanediamine in an organic solvent; and A method for producing a polymer compound, comprising the step of adding an acid catalyst to the above mixture.

8. In paragraph 7, A method further comprising the step of dispersing the polymer compound in an aqueous solution and performing ultrasonic treatment.

9. In paragraph 7, The above organic solvents are ethanol, methanol, isopropanol, t-butanol, tetrahydrofuran (THF), dimethylformamide (DMF), dichloromethane (CH 2 Cl 2 ), 1,2-dichloroethane (CH 2 ClCH 2 Cl), dimethyl sulfoxide (DMSO), t-butyl methyl ether (TBME), acetonitrile (ACN), diethyl ether, diphenyl ether, diisopropyl ether (DIPE), dimethyl acetamide (DMA), chlorobenzene, benzene, toluene, carbon tetrachloride (CCl 4 ), acetone, trifluoroacetic acid, chloroform (CHCl 3 ), pyridine and their aqueous solutions, or a mixed solution thereof.

10. In paragraph 7, A method wherein the acid catalyst comprises acetic acid, nitric acid, hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, perchloric acid, hydrofluoric acid or aluminum chloride.

11. In paragraph 7, A method wherein the above mixture is heated and stirred at 80°C for 16 hours.

12. In paragraph 8, A method wherein the above ultrasonic treatment is performed at 200 W for 10 minutes.

13. In paragraph 8, A method wherein the above ultrasonic treatment converts a polymer compound into nanoparticles.

14. A pharmaceutical composition for photodynamic therapy of glioblastoma, comprising a polymer compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient: [Chemical Formula 1] 15. In paragraph 14, A composition wherein the compound generates active oxygen species upon exposure to light.

16. In paragraph 15, A composition wherein the above active oxygen species is at least one selected from the group consisting of singlet oxygen, superoxide, hydrogen peroxide, amino acids, proteins, lipids, by-products of cell metabolism, and oxygen-containing active substances.

17. In paragraph 14, A composition wherein the compound specifically fluorescently labels glioblastoma.

18. In paragraph 14, A composition wherein the compound penetrates the blood-brain barrier by 0.1 to 20 μm.

19. In paragraph 14, A composition wherein the compound reduces tumor size.

20. A method for photodynamic treatment of glioblastoma, comprising the step of administering to a subject a pharmaceutically effective amount of a polymer compound represented by the following chemical formula 1. [Chemical Formula 1]

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