KR20190125007A - Photosensitizer, composition having the same, and method for photodynamic treatment - Google Patents

Abstract

The present invention relates to a photosensitizer which is a compound represented by chemical formula 1, a composition comprising the photosensitizer, and a photodynamic treatment method, wherein R, R^1 to R^10, and A are as defined in claim 1. The present invention can reduce cost, time, and patient pain in treating cancer.

Description

PHOTOSENSITIZER, COMPOSITION HAVING THE SAME, AND METHOD FOR PHOTODYNAMIC TREATMENT}

The present invention relates to a photosensitive agent, a composition comprising the photosensitive agent, and a photodynamic therapy method.

Photodynamic therapy is a treatment method that can minimize the pain of cancer patients by using photochemical reaction. After the photosensitive agent is administered to the patient, the photosensitive agent is irradiated with cancer tissue and the photosensitive agent reacts with reactive oxygen species through photochemical reaction. It is a method of oxidizing and killing biomolecules of cancer cells by generating species. In this method, the patient's physical burden is small and retreatment is possible within one month, and it can be combined with existing cancer treatment.

In general, the photosensing agents 'Photofrin' and 'Radachlorin', which are used in actual photodynamic therapy, are synthesized based on porphyrin dye, and these dyes are used for photodynamic therapy because the efficiency of producing reactive oxygen species in the water system is very low. Very strong light energy (> 200-300 J / cm ² ) should be applied.

Therefore, in order to minimize side effects caused by strong light energy and laser irradiation time, iridium complexes showing very high ROS production efficiency have been studied as photosensitizers for photodynamic therapy, but iridium complexes are blue (400-450 nm). ) Has an absorption range and the absorption coefficient is very low, resulting in an absolutely low oxygen production rate compared to the production efficiency of high oxygen species.

The present invention is to solve the problems described above, to provide a photosensitizer excellent in cancer cell killing properties at a selective binding to cancer cells and low light energy.

This invention provides the composition containing the photosensitive agent by this invention.

This invention provides the photodynamic therapy method using the composition by this invention.

The problem to be solved by the present invention is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.

According to one embodiment of the present invention, it relates to a photosensitive agent, which is a compound represented by the following Chemical Formula 1.

Wherein R and R ¹ to R ¹⁰ are each hydrogen, halogen, amide, cyano, hydroxyl, nitro, acyl, straight or branched chain alkyl of 1 to 30 carbon atoms, straight or branched chain eggs of 2 to 30 carbon atoms, respectively. Kenyl, straight or branched chain alkynyl having 2 to 30 carbon atoms, alkoxy having 1 to 30 carbon atoms, alkylcarbonyl having 1 to 30 carbon atoms and aryl having 2 to 20 carbon atoms, A is halogen, BF ₄ , BPh _{4 ,} PF ₆ , PCl ₆ , OAc, OTf, NO ₃ , ClO ₄ , SO ₄ , SCN, SbF ₆ And NTf ₂ .

According to an embodiment of the present invention, R and R ¹ to R ¹⁰ are each hydrogen, straight or branched chain alkyl of 1 to 20 carbon atoms, straight or branched chain alkenyl of 2 to 20 carbon atoms, and 2 to C carbon atoms. It may be selected from 20 straight or branched chain alkynyl.

According to an embodiment of the present invention, R and R ¹ to R ¹⁰ are each hydrogen, straight or branched chain alkyl of 1 to 10 carbon atoms, straight or branched chain alkenyl and 2 carbon atoms of 2 to 10 carbon atoms, respectively. To 10 straight or branched chain alkynyl may be selected.

According to one embodiment of the invention, the photosensitizer may be to generate active oxygen by irradiation of light energy of 200 J / cm ² or less.

According to one embodiment of the invention, the photosensitizer may be to generate active oxygen by irradiation of light energy of 10 J / cm ² or less.

According to an embodiment of the present invention, the photosensitive agent may have light emission characteristics and two-photon absorption characteristics in the wavelength region of 500 nm to 600 nm.

According to one embodiment of the invention, an active ingredient comprising at least one of the photosensitizers according to the invention; And a pharmaceutically acceptable carrier.

According to one embodiment of the invention, the composition may be for photodynamic therapy.

According to one embodiment of the invention, the composition may be for cancer treatment.

According to one embodiment of the invention, the composition, melanoma, skin cancer, colon cancer, pancreatic cancer, biliary tract cancer, neuroendocrine tumor, lung cancer, breast cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, gastric cancer, bladder cancer , Colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, head and neck cancer, thyroid cancer, parathyroid cancer and ureter cancer.

According to one embodiment of the invention, administering to a subject a composition for photodynamic therapy according to the invention; Allowing a predetermined time for the composition to be administered to accumulate in a target cell of a subject; And irradiating light energy to a target cell site of the subject. It relates to a photodynamic therapy method.

According to one embodiment of the present invention, the irradiating step may be to irradiate a total of less than 200 J / cm ² light energy for 10 seconds or more. According to one embodiment of the invention, the administering step may be to administer by oral or parenteral methods.

The present invention can provide a photosensitive agent capable of active oxygen species at a significantly lower light energy, higher productivity of active oxygen species, and excellent killing effect on cancer cells compared to conventional commercially available photosensitive agents. In addition, the photosensitizer can provide effective cancer cell killing ability even at a light energy of about 1 / 1000th lower than the light energy used in conventional photodynamic therapy, thereby reducing the cost, time, and pain of patients in treating cancer. Can be reduced.

Figure 1, according to an embodiment of the present invention, illustrates the concept of the intramolecular energy transfer process and photodynamic therapy mechanism of the photosensitive agent of the present invention, (a) acedan-iridium according to the present invention The intramolecular energy transfer process of the complex, (b) photodynamic therapy mechanisms using conventional photosensitive agents, and (c) photodynamic therapy mechanisms of acedan-iridium-complexes according to the present invention.
Figure 2 shows the absorption and photoluminescence spectra according to the evaluation example of the present invention, (a) the absorption spectrum of Ir-bxACD, Ir-COOH and bxACD-COOH, (b) Ir-bxACD, Ir-COOH and The light emission spectrum of bxACD-COOH and the light emission broad spectrum of (c) Ir-COOH and Ir-bxACD materials are shown.
Figure 3, according to the evaluation example of the present invention, the absorption spectrum of the ABDA and the absorbance attenuation according to time after the dye administration and irradiating light of Ir-COOH, bxACD-COOH and Ir-bxACD (compared to 0 minutes).
4 illustrates mitochondrial target motion using Ir-bxACD cancer cell (HeLa) imaging and Mitotracker, in accordance with an evaluation example of the present invention.
Figure 5 shows the cancer cell (HeLa) killing test of Ir-bxACD according to the evaluation example of the present invention,-Ir-bxACD / + light, + Ir-bxACD / + light, + Ir-bxACD /-light conditions DIC image (top), propidium iodide (middle) and DIC image and propidium iodide overlap image (bottom) are shown in FIG.
FIG. 6 shows a cancer cell (HeLa) killing test of Ir-bxACD according to an evaluation example of the present invention, and shows a DIC image (top) and a propidium iodide image (bottom).

Hereinafter, exemplary embodiments will be described in detail with reference to the accompanying drawings. However, various changes may be made to the embodiments so that the scope of the patent application is not limited or limited by these embodiments. It is to be understood that all changes, equivalents, and substitutes for the embodiments are included in the scope of rights.

The terminology used herein is for the purpose of description and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this specification, terms such as "comprise" or "have" are intended to indicate that there is a feature, number, step, action, component, part, or combination thereof described on the specification, and one or more other features. It is to be understood that the present invention does not exclude the possibility of the presence or the addition of numbers, steps, operations, components, components, or a combination thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art and shall not be construed in ideal or excessively formal meanings unless expressly defined in this application. Do not.

In addition, in the description with reference to the accompanying drawings, the same components regardless of reference numerals will be given the same reference numerals and duplicate description thereof will be omitted. In the following description of the embodiment, when it is determined that the detailed description of the related known technology may unnecessarily obscure the gist of the embodiment, the detailed description thereof will be omitted.

The present invention relates to a photosensitizer, and according to one embodiment of the invention, the photosensitizer is an acedan dye ((Acedan dye, energy donor, energy donor)) in an iridium complex (energy acceptor) Is a covalently bonded acedan-iri complex, by introducing an intramolecular energy transfer system by the binding of the acedan dye, thereby improving the absorption ability of the iridium complex, the active oxygen by light energy It is possible to improve species production.

According to one embodiment of the present invention, the photosensitizer is a compound represented by Formula 1 below, and may be an acedan-iridium complex in which an acedan dye and an iridium complex are combined.

[Formula 1]

Wherein R and R ¹ to R ¹⁰ are each hydrogen, halogen, amide, cyano, hydroxyl, nitro, acyl, straight or branched chain alkyl of 1 to 30 carbon atoms, straight or branched chain eggs of 2 to 30 carbon atoms, respectively. Kenyl, straight or branched chain alkynyl having 2 to 30 carbon atoms, alkoxy having 1 to 30 carbon atoms, alkylcarbonyl having 1 to 30 carbon atoms and aryl having 2 to 20 carbon atoms, A is halogen, BF ₄ , BPh _{4 ,} PF ₆ , PCl ₆ , OAc, OTf, NO ₃ , ClO ₄ , SO ₄ , SCN, SbF ₆ and NTf ₂ .

R and R ¹ to R ¹⁰ are each selected from hydrogen, straight or branched chain alkyl having 1 to 20 carbon atoms, straight or branched chain alkenyl having 2 to 20 carbon atoms, and straight or branched chain alkynyl having 2 to 20 carbon atoms, respectively. It may be. More preferably, R and R ¹ to R ¹⁰ are each straight or branched chain alkyl having 1 to 10 carbon atoms, straight or branched chain alkenyl having 2 to 10 carbon atoms and straight or branched chain alky having 2 to 10 carbon atoms, respectively. It may be selected from neil.

Here, R and R ¹ to R ¹⁰ may be independently selected as a functional group of one of the above-mentioned definitions according to the effect to be implemented through the compound of formula (1). In addition, R and R ¹ to R ¹⁰ may be selected from a combination of one or more functional groups (or elements) of the above-mentioned definitions depending on the effect to be realized through the compound of formula (1). At this time, the effect implemented according to the selected functional group may be implemented differently.

Referring to FIG. 1, FIG. 1 exemplarily illustrates the concept of an intramolecular energy transfer process and a photodynamic therapy mechanism of a photosensitive agent according to an embodiment of the present invention. In the acedan dye has a high absorption coefficient and a strong quantum efficiency (Quantum yield) and photoluminescence in the blue region (450-480 nm), and exhibits two-photon absorption characteristics, the light absorbed by the acedan dye Energy can be transferred to the iridium complex to enhance the low absorption coefficient of the iridium complex.

The photosensitizer is applied to photodynamic therapy (PDT), which can provide effective killing of target cells at extremely low light energy compared to conventional photodynamic therapy. That is, in the photodynamic therapy mechanism of FIG. 1 (b), the generation of reactive oxygen species is increased by using the light energy lower than that of the conventional photodynamic therapy according to the improvement of the absorbing ability of the iridium complex, and the effective killing of target cells. And / or inhibit growth / proliferation.

The photosensitizer, 200 J / cm ² or less; 100 J / cm ² or less; 50 J / cm ² or less; 10 J / cm ² or less; The generation of reactive oxygen species in the irradiation of light energy of more than 0 to 1 J / cm ² , the production amount of reactive oxygen species is excellent, can effectively provide the killing and / or growth / growth inhibition properties of the target tissue and / or cells. Can be.

The photosensitive agent may include a wavelength range of 500 nm to 600 nm; It has an absorbing region in the wavelength range of 550 nm to 580 nm and provides luminescent properties and two-photon absorption properties, so that imaging for selective binding and / or accumulation of target tissues and / or cells, for example, in vivo ) Or in vitro in vivo imaging may be possible.

The target tissues and cells are cancer tissues and cells, for example, melanoma, skin cancer, colon cancer, pancreatic cancer, biliary tract cancer, neuroendocrine tumor, lung cancer, breast cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer And gastric cancer, bladder cancer, colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, head and neck cancer, thyroid cancer, parathyroid cancer, and ureter cancer tissues and cells.

The present invention relates to a composition comprising a photosensitive agent according to the present invention. According to one embodiment of the present invention, the photosensitive agent may be applied to photodynamic therapy.

The composition is one or more of the photosensitizers according to the present invention; And a carrier; wherein, in the case where the photosensitive agent includes two or more kinds, the mixing ratio of the first photosensitive agent to the remaining photosensitive agent is effectively 1: 0.1 to 1 in order to effectively achieve selective binding and killing of the target tax. (w / w); Or 1: 0.1 to 0.5 (w / w).

The photosensitive agent is included as a pharmaceutically effective ingredient in the composition, and the "pharmaceutically effective amount" is an amount sufficient to achieve a therapeutic effect when the photosensitive agent according to the present invention is applied to a disease and / or photodynamic therapy. Means. For example, 0.001 to less than 100 parts by weight relative to 100 parts by weight of the composition; 0.001 part by weight to 50 parts by weight; Or 0.1 parts by weight to 10 parts by weight.

The carrier is applicable to photodynamic therapy and is a pharmaceutically acceptable carrier. For example, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl Cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.

The composition may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

The composition may be parenterally administered, for example intravenous administration, intraperitoneal administration, intramuscular administration, subcutaneous administration or topical administration. In addition, oral administration, rectal administration, inhalation administration, nasal administration and the like may be possible.

Suitable dosages of the compositions vary depending on factors such as formulation method, mode of administration, patient's age, weight, sex, degree of disease symptom, food, time of administration, route of administration, rate of excretion, and reaction sensitivity. The skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment.

The composition may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art, or It can be prepared by incorporation into a dose container. The formulations may then be in the form of solutions, suspensions or emulsions in oils or aqueous media, and may further comprise dispersants or stabilizers.

The composition is for the treatment of cancer, for example, melanoma, skin cancer, colon cancer, pancreatic cancer, biliary tract cancer, neuroendocrine tumor, lung cancer, breast cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, gastric cancer, bladder cancer , Colon cancer, cervical cancer, brain cancer, prostate cancer, bone cancer, head and neck cancer, thyroid cancer, parathyroid cancer and ureter cancer.

The present invention relates to a photodynamic therapy method using a photosensitive agent according to the present invention. According to one embodiment of the present invention, the photodynamic therapy method is administered to a subject with a composition for photodynamic therapy. Making; Allowing a predetermined time for the composition to be administered to accumulate in a target cell of a subject; And irradiating light energy to a target cell site of the subject. It may include.

The method may further include imaging the target tissue for identifying the position of the target cell of the subject after allowing the predetermined time. This can be detected by visualizing the fluorescence or luminescence image by light irradiation to identify the target tissue (or cells).

The irradiating step may include at least 1 second; 10 seconds or more; Alternatively, a total of 200 J / cm ² or less light energy may be irradiated for 1 second to 5 hours. The administering step may be administered by oral or parenteral methods.

The “subject” is a mammal without limitation, and may include, for example, humans, mice, rats, guinea pigs, dogs, cats, horses, cattle, pigs, monkeys, chimpanzees, bears, rabbits, baboons or rhesus monkeys, and the like. Can be.

Although the present invention will be described with reference to preferred embodiments, the present invention is not limited thereto, and the present invention is not limited to the scope of the present invention as defined in the following claims, the detailed description of the invention, and the accompanying drawings. Various modifications and variations can be made in the present invention.

Scheme 1

Example

According to Scheme 1, acedan dye (4, bxACD-COOH) was synthesized, and a linkage and acedan-iridium complex (11, Ir-bxACD) were synthesized.

(1) Compound 1

6- Bromo -N- Butylnaphthalene 2-ami (6- bromo -N- butylnaphthalen -2-amine) synthesis

6-bromo-2-naphthylamine (6-bromo-2-naphthylamine, 2.00 g, 9.00 mmol), Na ₂ S ₂ O ₅ (3.07 g, 10.8 mmol), butylamine (5 ml, 45.0 mmol) ) Was dissolved in water (10ml) and refluxed for 2 days. The crude product was then cooled to room temperature and poured into 2N NaOH solution. After placing the material at 0 ° C. for 1 hour, the resulting solid was filtered through a filter and washed with cold water to obtain Chemical 1.

(2) compound 2

Tert -Butyl-N- (6- Bromonaphthalene 2-yl) -N- Butyl Glycinate ( tert -butyl N- (6-bromonaphthalen-2-yl) -N-butylglycinate)

After dissolving Compound 1 (200 mg, 0.720 mmol), proton sponge (169 mg, 0.790 mmol), and tert-butyl bromoacetate (384 μl, 2.59 mmol) in 20 ml of MeCN, 12 hours Was refluxed for a while. After lowering the temperature to room temperature, the product was extracted into the organic layer using water and ethyl acetate, and Compound 2 was purified by column chromatography (Hex 5: EA 1).

(3) compound 3

Tert -Butyl N- (6- Benzoxazole -2-yl) naphthalen-2-yl) -N- Butyl Glycinate ( tert -butyl N- (6- (benzoxazol-2-yl) naphthalen-2-yl) -N-butylglycinate)

Compound 2 (280 mg, 0.716 mmol), benzoxazole (102 mg, 0.86 mmol), Pd (IImg, 0.036 mmol), PPh ₃ (19 mg, 0.072 mmol), CuI (27 mg, 0.143 mmol) and CsCO ₃ (280 mg, 0.86 mmol) was dissolved in 10 ml of DMF without water and oxygen, and then refluxed at 140 ° C. for 12 hours in an Ar environment. The solid of the crude product (Crude product) was filtered through a filter, and the product was extracted into the organic layer using water and ethyl acetate. Compound 3 was then purified via column chromatography (Hex 5: EA 1).

(4) compound 4

N- (6- ( Benzoxazole -2-yl) naphthalen-2-yl) -N- Butyl Glycine ( bxACD - COOH , N- (6- (benzoxazol-2-yl) naphthalen-2-yl) -N-butylglycine)

Compound 3 (100 mg, 0.232 mmol) and CF ₃ COOH (1.5 ml) were dissolved in deoxygenated DCM (10 ml) and stirred at room temperature for 24 hours. Then, the solvent was evaporated using a rotary distillation and washed with hexane (Hexane) to filter out the solid (Compound 4) through a filter.

(5) Compound 5

6- (4'- Methyl [[2,2'-bipyridin] -4-yl) hexanenitrile (6- (4'-methyl- [2,2'-bipyridin] -4-yl) hexanenitrile)

Diisopropylamine (diisopropylamine, 1.79 ml, 12.7 mmol) was dissolved in THF from which water was removed, and n-BuLi (2.5M, 4.65 ml) was added dropwise at -78 ° C. After stirring for 20 minutes, 4,4 '′-dimethyl-2,2 ′ ′-dipyridyl (4,4 ′ ′-Dimethyl-2,2 ′ ′-dipyridyl, 2 g, 10.6 mmol) was dissolved in THF. Drop by drop. After further stirring for 1 hour and 30 minutes, 5-bromovaleronitrile (5-bromovaleronitrile, 1.5 ml, 12.7 mmol) was added thereto, and the mixture was stirred at 0 ° C. for 2 hours. 10 ml of water was added to the mixture, neutralized with 1N aqueous HCl solution, and extracted with water and ethyl acetate. Compound 5 was then purified via column chromatography (Hex 10: EA 1).

(6) compound 6

6- (4'-Methyl- [2,2'-bipyridin] -4-yl) hexanoic acid (6- (4'-methyl- [2,2'- bipyridin ] -4-yl) hexanoic acid)

Compound 5 (570 mg, 2.15 mmol) was added to Conc. It was put in HCl (35 ~ 37%) and refluxed at 100 ℃. After lowering the temperature of the mixture to room temperature, the pH was adjusted to 4 to 4.5 with 6N NaOH aqueous solution, and the product was extracted into the organic layer with chloroform and water. Compound 6 was obtained by evaporating all chloroforms using a rotary distillation machine.

(7) Compound 7

Tert - Butyl (2- (6- (4'-methyl-) [2,2'- Bipyridine ] -4-day) Hexane Amido )ethyl) Kabamei Tert-butyl (2- (6- (4'-methyl- [2,2'- bipyridin ] -4-yl) hexanamido) ethyl) carbamate)

Compound 6 (300 mg, 1.06 mmol), HATU (361 mg, 0.95 mmol) and diisopropylethylamine (Diisopropylethylamine, DIPEA, 409 mg, 3.17 mmol) were dissolved in DMF and stirred for 15 minutes. N-boc-ethylenediamine (N-boc-ethylenediamine, 253 mg, 1.58 mmol) was slowly added thereto, followed by stirring at room temperature for 5 hours. Then the product was extracted into the organic layer using ethyl acetate and water and the solvent was evaporated using rotary distillation. Compound 7 was purified by column chromatography (Chloroform 10: MeOH 1).

(8) compound 8

N- (2- Aminoethyl ) -6- (4'-methyl- [2,2'-bipyridin] -4-yl) hexanamide (N- (2-aminoethyl) -6- (4'-methyl- [2,2'-bipyridin] -4-yl) hexanamide)

Compound 7 (409 mg) was dissolved in DCM and slowly injected with CF ₃ COOH (8 ml). After stirring for 2 hours, 10 ml of toluene was added and all solvents were evaporated in a rotary distillation to obtain Compound 8.

(9) Compound 9

(N- (2- (2-((6- ( Benzo [d] oxazole -2-yl) naphthalen-2-yl) (butyl) amino) Acetamido ) Ethyl) -6- (4'-methyl- [2,2'-bipyridin] -4-yl) hexanamide) (naphthalen-2-yl) (butyl) amino) acetamido) ethyl) -6- (4 '-methyl- [2,2'-bipyridin] -4-yl) hexanamide)

Compound 4 (139 mg, 0.372 mmol), HATU (159 mg, 0.418 mmol), DIPEA (180 mg, 1.392 mmol) was dissolved in DMF and stirred for 15 minutes. Then compound 8 (204 mg, 0.464 mmol) was slowly injected and stirred at room temperature for 5 hours. The mixture was extracted into an organic layer using ethyl acetate and water and compound 9 was obtained by column chromatography.

(10) Compound 11

( Ir - bxACD , Acedan Iridium complex)

Compound 10 is described in “ J. Am. Chem . Soc ., 2016, 138 (34), 10968-10977) ”. Compound 10 and compound 9 were dissolved in DCM: MeOH = 1: 1 (v / v) solvent, and then refluxed for 12 hours. After evaporation of all solvents using a rotary distillator, the product was dissolved in ethanol and the remaining reactants were removed through a filter. The ethanol was evaporated again using a condenser, and then solidified with DCM and hexane, and a compound 11 (Ir-bxACD) was obtained using a filter.

Evaluation example

Photochemical Properties and Intramolecular Energy Transfer

Absorption and photoluminescence spectra of the Ir-bxACD material and Ir-COOH and bxACD-COOH of Examples were measured and shown in FIG. 2. The high efficiency intramolecular energy transfer (98.9%) of the Ir-bxACD material can be seen in the absorption and photoluminescence spectra of FIG. 2.

In addition, the orange spectrum of Ir-bxACD in the absorption spectrum appears as a linear sum of the spectrums of the remaining Ir-COOH and bxACD-COOH, which means that the acedan and iridium portions of Ir-bxACD absorb light independently of each other. It can be seen that the light absorption ability of Ir-bxACD is improved compared to the existing iridium complex (Ir-COOH).

In addition, the light emission spectrum can confirm whether the acedan portion of Ir-bxACD can effectively transmit light energy absorbed independently to the iridium complex. In other words, the spectrum of Ir-bxACD is much smaller than that of acedan dye (bxACD-COOH) in the middle photoluminescence graph. Instead of stabilizing, an energy transfer occurs to the iridium complex, which can be confirmed by the tendency of the light emission spectrum of Ir-bxACD to increase compared to Ir-COOH.

< Singlet  Oxygen Generation Efficiency>

A comparison of singlet oxygen production by ABDA (Anthracenediyl-bis (methylene) dimalonic acid) assay is shown in FIG. 3. ABDA reacts rapidly with singlet oxygen, causing the anthracene central molecule to collapse easily. Therefore, the singlet oxygen generating ability can be compared by the degree of attenuation of the absorption spectrum at 400 nm indicated by the anthracene core.

In FIG. 3, when the acedan dye (bxACD-COOH) was added and irradiated with light, the absorption spectrum of the ABDA did not attenuate at all, and the absorption spectrum of the conventional iridium complex (Ir-COOH) was somewhat attenuated while the iridium-acedan In the case of the complex (Ir-bxACD), it can be seen that the absorbance spectrum is attenuated very rapidly, indicating that the Ir-bxACD generates most singlet oxygen most effectively when absorbed light. Experimental conditions of FIG. 3: [ABDA] = 66.7 μΜ, [Dye] = 6.67 μΜ in DMSO: H- ₂ O solution (1:99 = v / v) Light source: Xe white Lamp (0.4 sun).

< Cancer cell Confocal Imaging  >

Ir-bxACD was applied to actual live cancer cells to measure cancer cell coordination imaging and are shown in FIG. 4. In Figure 4 it can be seen that the acedan-irilium-complex of the present invention Ir-bxAC acts in the mitochondria of cancer cells. Experimental conditions of FIG. 4: Cell line = HeLa, [Ir-bxACD] = 2 μΜ, 60 min incubation, [Mitotracker] = 200 nM, 30 min incubation. Instrumant: FV-1000 confocal microscopy.

<In vitro cancer cell killing experiment (12 J / cm ² )>

After administering Ir-bxACD material to cancer cells, dead cancer cells were stained with propidium iodide to confirm that the cancer cells were effectively killed when irradiated with light. The results are shown in FIG. 5 (Experimental conditions: Cell line = HeLa, [Ir-bxACD] = 4 μM, 100 min incubation, then 1 sun light irradiation for 2 min (Xe white lamp 12 J / cm ² ), 5 hour incubation. [PI] = 1.5 μΜ, 30 min).

In FIG. 5, irradiated with only light without administering Ir-bxACD (Xe white lamp, 12 J / cm ² ) and in two cases irradiated with Ir-bxACD but not irradiated with light (left and right in FIG. 5). Was able to confirm that it was alive while maintaining its normal shape, but when Ir-bxACD was administered and irradiated with light (Xe white lamp, 12 J / cm ² ), almost all cells were killed to give red fluorescence of propidium iodide. It was. Therefore, it can be seen that both light and Ir-bxACD are essential factors for cancer cell death.

<In vitro cancer cell killing experiment (0.24 J / cm ² )>

Irradiation with LED blue light for 1 minute after irradiation with Ir-COOH, Ir-bxACD and propidium iodide to confirm that cancer cells can be effectively killed with very weak light energy of less than 0.3 J / cm ² / cm ² ), and imaging was performed at 30 minute intervals. The result is shown in FIG. Experimental conditions: Cell line = HeLa, [Ir-bxACD] = 4 μΜ, 120 min incubation, then Blue LED light irradiation for 1 min (0.24 J / cm ² ). [PI] = 1.5 μM, 30 min incubation

When irradiated with light in FIG. 6 for 60 minutes, Ir-bxACD showed fluorescence of red iodide propidium in almost all cells, whereas in Ir-COOH, most of the cells were alive. This means that even at ¹ / 1000th of the light energy (0.3 J / cm ² ) of the light energy (200-300 J / cm ² ) used in actual photodynamic therapy, the acedan-iridium complex is much more effective than the conventional iridium complex. It is the ability to die. Therefore, Ir-bxACD with high singlet oxygen production efficiency can induce effective cell death with less light energy than Ir-COOH, minimize the side effects of cancer patients due to strong light energy in actual photodynamic therapy, The treatment time can be reduced.

The present invention provides a conventional optical dynamic by introducing an "intramolecular energy transfer" by covalently connecting an acedan dye (energy donor) to an iridium complex (energy acceptor). It provides a photosensitive agent for photodynamic therapy, a composition comprising the same, and a photodynamic therapy method using the same, which can effectively kill cancer cells even at very low light energy (for example, <0.3 J / cm ² ) compared to the treatment. can do.

Although the embodiments have been described with reference to the accompanying drawings, those skilled in the art may apply various technical modifications and variations based on the above. For example, the described techniques may be performed in a different order than the described method, and / or components of the described systems, structures, devices, circuits, etc. may be combined or combined in a different form than the described method, or other components Or even if replaced or substituted by equivalents, an appropriate result can be achieved.

Therefore, other implementations, other embodiments, and equivalents to the claims are within the scope of the following claims.

Claims (13)

A photosensitive agent, which is a compound represented by the following formula (1).

Wherein R and R ¹ to R ¹⁰ are each hydrogen, halogen, amide, cyano, hydroxyl, nitro, acyl, straight or branched chain alkyl of 1 to 30 carbon atoms, straight or branched chain of 2 to 30 carbon atoms, respectively. Alkenyl, straight or branched chain alkynyl having 2 to 30 carbon atoms, alkoxy having 1 to 30 carbon atoms, alkylcarbonyl having 1 to 30 carbon atoms and aryl having 2 to 20 carbon atoms,
A is halogen, BF ₄ , BPh _{4 ,} PF ₆ , PCl ₆ , OAc, OTf, NO ₃ , ClO ₄ , SO ₄ , SCN, SbF ₆ And NTf ₂ ).
The method of claim 1,
R and R ¹ to R ¹⁰ are each selected from hydrogen, straight or branched chain alkyl having 1 to 20 carbon atoms, straight or branched chain alkenyl having 2 to 20 carbon atoms, and straight or branched chain alkynyl having 2 to 20 carbon atoms, respectively. Photosensitive agent that becomes.
The method of claim 1,
R and R ¹ to R ¹⁰ are each selected from hydrogen, straight or branched chain alkyl of 1 to 10 carbon atoms, straight or branched chain alkenyl of 2 to 10 carbon atoms, and straight or branched chain alkynyl of 2 to 10 carbon atoms, respectively. Photosensitive agent that becomes.
The method of claim 1,
The photosensitizer is a photosensitive agent that generates free radicals upon irradiation of light energy of 200 J / cm ² or less.
The method of claim 1,
The photosensitizer is a photosensitive agent that generates free radicals under irradiation of light energy of 10 J / cm ² or less.
The method of claim 1,
The photosensitive agent is one having a light emission characteristic and a two-photon absorption characteristics in the wavelength region of 500 nm to 600 nm.
An active ingredient comprising at least one of the photosensitizers of claim 1; And
Pharmaceutically acceptable carriers;
Including,
Composition.
The method of claim 7, wherein
The composition is a composition for photodynamic therapy.
The method of claim 7, wherein
The composition is that for cancer treatment.
The method of claim 7, wherein
The composition, melanoma, skin cancer, colon cancer, pancreatic cancer, biliary tract cancer, neuroendocrine tumor, lung cancer, breast cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, gastric cancer, bladder cancer, colon cancer, cervical cancer, brain cancer, prostate Cancer, bone cancer, head and neck cancer, thyroid cancer, parathyroid cancer and ureter cancer treatment, the composition.
Administering to the subject a composition for photodynamic therapy of claim 7;
Allowing a predetermined time for the composition to be administered to accumulate in a target cell of a subject; And
Irradiating light energy to a target cell site of the subject;
Including,
Photodynamic therapy method.
The method of claim 11,
The irradiating step is to irradiate a total of 200 J / cm ² or less light energy for at least 10 seconds, photodynamic therapy method.
The method of claim 11,
The administering step, orally or parenterally administered, photodynamic therapy method.

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