Degradable near-infrared photosensitizers and methods for preparing and using same - Patents.com

Abstract

The present invention discloses a decomposable near-infrared photosensitizer, its preparation method and application, and belongs to the technical field of photosensitizers. The decomposable near-infrared photosensitizer is any one selected from a porphyrin compound, its pharma- ceutically acceptable salt, solvate, non-covalent complex, complex, and prodrug, and the structure of the porphyrin compound is as shown in Formula 1. The present invention discloses a preparation method and application of the decomposable near-infrared photosensitizer. According to the decomposable near-infrared photosensitizer of the present invention, active oxygen is generated and decomposed by light irradiation, and after the end of light irradiation, the photosensitizer is completely converted into a low-toxicity substance that has no photoactivity, effectively solving the problem of remaining photosensitizers in the prior art. [Formula 1] TIFF2024537897000026.tif56170

Description

[Technical Field]

[0001] The present invention relates to a decomposable near-infrared photosensitizer, as well as its preparation method and use, which belongs to the technical field of photosensitizers. [Background technology]

[0002] Cancer is one of the most serious diseases threatening human health, with an extremely high mortality rate and threatening the safety of human life. Compared to current major cancer treatment methods, such as surgical resection, radiation therapy, and chemotherapy, photodynamic therapy (PDT) is a non-invasive treatment with advantages in terms of treatment time, therapeutic efficacy, drug resistance, toxicity, and side effects. It has shown great potential in the fields of cancer, tumors, skin diseases, and antibacterial treatment. The mechanism of action of PDT is as follows: a photosensitizer is injected into the body, transported to the lesion via the blood circulation, and then accumulated in the lesion. The lesion is then locally irradiated with light of a specific wavelength, which excites the photosensitizer and releases reactive oxygen species (ROS), which directly damage and destroy blood vessels in the lesioned tissue and induce immune stress, thereby killing the diseased cells and achieving the therapeutic goal. Photodynamic therapy is classified into type I or type II depending on the mechanism by which reactive oxygen species are generated. Type I photodynamic reaction occurs when radicals are generated from photosensitizers by the transfer of electrons or protons, and then the highly active radicals interact with molecular oxygen to generate superoxide anions, which then react to generate reactive oxygen species such as hydroxyl radicals and hydrogen peroxide. The damage caused by this is called type I damage. Type II photodynamic reaction occurs when the photosensitizer transfers the energy of its excited state to surrounding oxygen molecules, causing them to become singlet oxygen, and the damage caused by this is called type II damage.

[0003] The effectiveness of photodynamic therapy depends primarily on the properties of the photosensitizer. Photosensitizers have strong phototoxicity but no toxicity or side effects to the body. In recent years, the number of photosensitizers has gradually increased, and there are now three generations. First-generation photosensitizers are primarily porphyrin-based mixtures. Second-generation photosensitizers refer to synthetic compounds that combine porphyrin derivative monomers with different structural types and active molecules. Third-generation photosensitizers are targeted photosensitizers obtained by combining photosensitizers with monoclonal antibodies or other small bioactive molecules such as steroids, lipids, peptides, nucleosides, and nucleotides. However, the scope of clinical application of photodynamic therapy is still limited, mainly due to the following reasons: (1) most photosensitizers still require short-wavelength excitation, but short-wavelength light has insufficient penetration depth, making it difficult to effectively irradiate deep lesions; (2) due to its high oxygen dependency, the effectiveness of photodynamic therapy on anoxic solid tumors is limited; (3) it is poorly water-soluble, prone to aggregation in the body, and lacks specificity; and (4) the structure of photosensitizers is stable, and their decomposition and metabolism in the body is slow.

[0004] Conventional photosensitizers have drawbacks such as limited treatment depth, high oxygen dependency, and poor tumor targeting, which severely limit the therapeutic effect of photodynamic therapy in clinical applications. Furthermore, after photodynamic therapy, the remaining photosensitizer in the body often takes a long time to be completely metabolized. When exposed to light, the remaining photosensitizer continues to generate reactive oxygen species, exhibiting strong phototoxicity and causing some damage to tissues and cells. Therefore, patients are required to avoid light for a long period of time after treatment. Living in constant darkness and fear for several months can have serious consequences on the patient's physical and mental health. Therefore, timely deactivation of the photosensitizer after photodynamic therapy to avoid safety issues caused by the remaining photosensitizer is a challenge in the clinical application of photodynamic therapy. Summary of the Invention [Problem to be solved by the invention]

[0005] In view of this, it is necessary to provide a new decomposable near-infrared photosensitizer, its preparation method and application to make up for the shortcomings of the prior art. [Means for solving the problem]

[0006] A first object of the present invention is to provide a decomposable near-infrared photosensitizer.

[0007] The present invention has the following configuration for solving the above technical problems.

[0008] A decomposable near-infrared photosensitizer, which is any one selected from a porphyrin compound, a pharmaceutically acceptable salt thereof, a solvate thereof, a non-covalent complex thereof, a complex thereof, and a prodrug thereof, wherein the structure of the porphyrin compound is as shown in Formula 1, [ka] During the ceremony, [ka] are each independently [ka] Any one selected from Ar1 and Ar2 are both substituted or unsubstituted phenyl groups, R1, R2, R3, R4, R5, R6, R7, and R8 each independently represent hydrogen, halogen, a nitro group, a hydroxyl group, an amino group, a sulfanyl group, a carboxyl group, a sulfonic acid group, a phosphate group, a cyano group, an amide group, an amino group substituted with a C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C 1-8any one selected from an alkoxy group, a substituted mercapto group, a C1-C8 alkyl phosphate group, a C1-C8 alkyl carboxyl group, a C1-C8 alkyl sulfonate group, a C3-C6 alkenyl alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl alkyl group, a C2-C6 alkynyl group, a C2-C6 alkenyloxy group, and a C2-C6 alkynyloxy group; M is a metal ion and is any one selected from Mg, Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, Ga, Al, Pd, and Pt.

[0009] The mechanism of the decomposable near-infrared photosensitizer of the present invention will be explained as follows.

[0010] The present invention modifies the structure of porphyrin compounds to adjust the photophysical and photochemical properties of the compounds, thereby realizing the near-infrared absorption and luminescence properties of the photosensitizer molecules, as well as the efficacy of type I and type II photodynamic therapy. The lactone unit in the molecular structure of the photosensitizer can react with active oxygen species, and when activated by light irradiation, it is converted into a non-photoactive, low-toxicity substance, and the photosensitizer is gradually decomposed.

[0011] The decomposable near-infrared photosensitizer of the present invention has the following advantageous effects.

[0012] 1. The decomposable near-infrared photosensitizer of the present invention generates active oxygen upon irradiation with light and is decomposed at the same time. After the end of irradiation, it is completely converted into a non-photoactive, low-toxic substance, thereby effectively solving the problem of residual photosensitizers in the prior art. 2. The decomposable near-infrared photosensitizer of the present invention has an absorption spectrum in the near-infrared region (700 nm to 900 nm), a large extinction coefficient, and a large penetration depth of excitation light into tissue, making it useful for reducing damage to tissue caused by light sources. 3. The decomposable near-infrared photosensitizer of the present invention has strong fluorescence, so that it can be used for fluorescent labeling to monitor the decomposition degree of the photosensitizer in real time. 4. The degradable near-infrared photosensitizer of the present invention exhibits high phototoxicity at the cellular and biological levels, and can be used as an active ingredient in the preparation of photodynamic therapeutic drugs.

[0013] Based on the above configuration, the present invention can be improved as follows.

[0014] Furthermore, Ar1 and Ar2 are each independently [ka] and In the formula, R9, R 10 , R 11 , R 12 , and R 13 are each independently any one selected from hydrogen, halogen, a nitro group, a hydroxyl group, an amino group, a sulfanyl group, a carboxyl group, a sulfonic acid group, a phosphate group, a cyano group, an amido group, an amino group substituted with a C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkoxy group, a substituted thio group, a C1-C8 alkyl phosphate group, a C1-C8 alkyl carboxyl group, a C1-C8 alkylsulfonic acid group, a C3-C6 alkenylalkyl group, a C2-C6 alkenyl group, a C2-C6 alkynylalkyl group, a C2-C6 alkynyl group, a C2-C6 alkenyloxy group, and a C2-C6 alkynyloxy group.

[0015] Another advantage of the above configuration is that the solubility and reactivity of the photosensitizer can be adjusted by different substituents to achieve biocompatibility modification and functional change to meet different application needs.

[0016] Furthermore, the metal ion is any one selected from Mg, Zn, Cd, Al and Ga.

[0017] The advantageous effects of the above additional configuration include controlling the efficacy of photosensitizers that generate fluorescence and active oxygen, improving the stability of the photosensitizers, and further reducing the dark toxicity of the photosensitizers, thereby improving biological safety.

[0018] Furthermore, the specific structure of the porphyrin compound is [ka] TIFF0007800874000006.tif53170 It is any one selected from the above.

[0019] A second object of the present invention is to provide a method for preparing the decomposable near-infrared photosensitizer.

[0020] The present invention is configured to solve the above technical problems as follows: A method for preparing the decomposable near-infrared photosensitizer, comprising the following steps 1 to 3: In step 1 of preparing the first intermediate product, 100 mg of porphodilactone and an alkali equivalent to 30 to 50 times the amount of the porphodilactone are dissolved in 5 mL to 20 mL of a first solvent in a hydrothermal reaction vessel, and the mixture is reacted at 80°C to 200°C for 24 hours to 72 hours to obtain the first intermediate product, and the reaction formula is as follows: [ka] In step 2 for preparing the second intermediate product, 100 mg of the first intermediate product obtained in step 1 is subjected to a water-soluble modification reaction in 5 mL to 50 mL of a second solvent to obtain the second intermediate product, and the reaction formula is as follows: [ka] In step 3 for preparing the target product, 100 mg of the second intermediate product obtained in step 2 and a metal salt equivalent to 10 times the amount of the second intermediate product are dissolved in 20 to 100 mL of a third solvent, and reacted at 20 to 80°C for 2 to 16 hours to obtain the target product shown in formula 1, and the reaction formula is as follows: [ka]

[0021] The mechanism of the method for preparing the decomposable near-infrared photosensitizer of the present invention is as follows.

[0022] In step 1 of the present invention, porphodilactone is used as a substrate, and one lactone group is converted into a cyclic conjugated structure under alkaline conditions to increase the degree of conjugation. The fluorine atom at the ortho position of the phenyl group in porphodilactone can undergo a nucleophilic substitution reaction, and the two hydroxyl groups after the ring-opening of the lactone group can attack the adjacent fluorine atom under alkaline conditions to form two new five-membered rings, stabilizing the rigid structure of the large ring. The chemical reaction scheme is as follows: [ka] is.

[0023] In step 2 of the present invention, R1 to R 13 Active groups such as amino, hydroxyl, cyano, and mercapto groups are introduced into the hydroxyl group, and further reactive modification is carried out to obtain water-soluble molecules with structures such as quaternary ammonium, sulfonic acid, carboxyl, and glycosyl groups.

[0024] In step 3 of the present invention, the first and second intermediate products have good coordination functions as macrocyclic tetrapyrrole ligands, and are therefore capable of chelating various metal ions. By selecting different metal ions, target products with different central metals can be obtained under heating conditions.

[0025] The advantageous effects of the method for preparing the decomposable near-infrared photosensitizer of the present invention are as follows:

[0026] 1. The present invention has a simple overall synthesis route, high yield, and is easy to purify by recrystallization. 2. The reaction conditions of the present invention are mild, the reaction is safe, there is no outflow of harmful substances, and pollution is reduced. 3. The present invention is low cost, easy to operate, has broad market prospects, and is suitable for scale-up and expanded application.

[0027] Based on the above configuration, the present invention can be improved as follows.

[0028] Furthermore, in step 1, the alkali is any one or a mixture of two or more selected from sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, triethylamine, N,N-diisopropylethylamine, sodium methoxide, and potassium tert-butoxide.

[0029] Another advantageous effect of the above configuration is that the use of strong alkali is avoided, improving the safety of the reaction operation.

[0030] Furthermore, the alkali is any one or a mixture of two or more selected from potassium carbonate, sodium carbonate, triethylamine, and N,N-diisopropylethylamine.

[0031] Furthermore, in step 1, the first solvent is any one or a mixture of two or more selected from water, methanol, ethanol, acetonitrile, acetone, dimethyl sulfoxide, N,N-dimethylformamide, and tetrahydrofuran.

[0032] Furthermore, the first solvent is a solvent in which water and acetonitrile are mixed in a volume ratio of 1:(2 to 10), or a solvent in which water and tetrahydrofuran are mixed in a volume ratio of 1:(2 to 10).

[0033] A further advantageous effect of the above configuration is that by employing the above parameters, the yield of the reaction can be improved.

[0034] Furthermore, the first solvent is a solvent in which water and acetonitrile are mixed in a volume ratio of 1:(5-10), or a solvent in which water and tetrahydrofuran are mixed in a volume ratio of 1:(5-10).

[0035] Furthermore, in step 1, the reaction temperature is set to 100° C. to 150° C., and the reaction time is set to 48 hours to 72 hours.

[0036] As a further advantageous effect of the above configuration, by adopting the above reaction parameters, the occurrence of side reactions is reduced, post-treatment is facilitated, and the reaction yield can be improved.

[0037] Furthermore, in step 2, the second solvent is any one or a mixture of two or more selected from dimethyl sulfoxide, N,N-dimethylformamide, tetrahydrofuran, ethyl acetate, trimethyl phosphate, triethyl phosphate, and hexanol.

[0038] As a further advantageous effect of the above configuration, by using the above-mentioned second solvent, post-treatment becomes easier and the reaction yield improves.

[0039] Furthermore, the second solvent is any one or a mixture of two or more selected from N,N-dimethylformamide, tetrahydrofuran, ethyl acetate, and trimethyl phosphate.

[0040] Furthermore, in step 3, the metal salt is any one selected from magnesium salts, chromium salts, manganese salts, iron salts, cobalt salts, nickel salts, copper salts, zinc salts, cadmium salts, gallium salts, aluminum salts, palladium salts, and lead salts.

[0041] The advantageous effect of the above further configuration is that the solubility of the metal salt in the third solvent is good, and the structure of the obtained target product is stable.

[0042] Furthermore, the metal salt is any one selected from the group consisting of magnesium salts, zinc salts, cadmium salts, gallium salts, and aluminum salts.

[0043] Furthermore, the metal salt is any one selected from acetate, chloride, sulfate, nitrate, and phosphate.

[0044] Furthermore, the metal salt is any one selected from acetate, chloride, and nitrate.

[0045] Furthermore, in step 3, the third solvent is any one or a mixture of two or more selected from water, methanol, ethanol, acetonitrile, acetone, dimethyl sulfoxide, N,N-dimethylformamide, tetrahydrofuran, chloroform, and dichloromethane.

[0046] The advantageous effects of the above further configuration include increasing the solubility of the metal salt in the solvent, reducing the amount of solvent used, and simplifying post-reaction treatment.

[0047] Furthermore, the third solvent is any one or a mixture of two or more selected from methanol, ethanol, acetonitrile, and acetone.

[0048] Furthermore, the third solvent is methanol or ethanol.

[0049] Furthermore, in step 3, the reaction temperature is set to 20°C to 60°C, and the reaction time is set to 3 hours to 5 hours.

[0050] The additional advantages of the above configuration include shortening the reaction time and improving the conversion of the reaction.

[0051] A third object of the present invention is to provide applications of the decomposable near-infrared photosensitizer.

[0052] The present invention has the following configuration for solving the above technical problems.

[0053] The application of said decomposable near-infrared photosensitizer as an active ingredient in the preparation of photodynamic therapeutic drugs.

[0054] The beneficial effect of applying the degradable near-infrared photosensitizer of the present invention is that the degradability characteristics of the degradable near-infrared photosensitizer will lead to the development of a new metabolic model for photosensitized drugs, which is expected to lead to the realization of a new photodynamic therapy model, and is of positive significance from both pharmaceutical and social perspectives.

[0055] Based on the above configuration, the present invention can be improved as follows.

[0056] Furthermore, the photodynamic therapeutic drug comprises the decomposable near-infrared photosensitizer and a pharmaceutically acceptable adjuvant, and its dosage form is any one selected from powder, injection, and emulsion.

[0057] The advantageous effects of the above-mentioned configuration include the ability to prepare photodynamic therapeutic drugs in various dosage forms and for various administration routes, and the degradable near-infrared photosensitizer in the photodynamic therapeutic drug can be concentrated at the affected site through diffusion, circulation, or targeted administration, resulting in rapid efficacy, accurate drug dosage, and reliable action.

[0058] The preferred routes of administration are injection and topical administration. Injection may be intradermal, subcutaneous, intramuscular, topical or intravenous, while topical administration includes administration to the affected area by means of compress, rubbing or painting.

[0059] Furthermore, the auxiliary agent is any one or a mixture of two or more selected from a solvent, an emulsifier, an isotonicity agent, a surfactant, and an antioxidant.

[0060] The beneficial effects of the above additional configuration include the ability to use the above auxiliary agents to prepare drugs in various dosage forms, enhance the stability of the photodynamic therapeutic agent, adjust the action of the photodynamic therapeutic agent, and improve biocompatibility. The above auxiliary agents should all be inert ingredients that are compatible with the administration route or mode and have no toxic effects on the human body.

[0061] Furthermore, the solvent is any one selected from ethanol, propylene glycol, N,N-dimethylformamide, and dimethyl sulfoxide.

[0062] The above further advantageous effect is to promote the solubility of the drug in different dosage forms, allowing the dosage of the active ingredient to be 0.01 mg to 20 g per unit dosage form of the photodynamic therapeutic agent.

[0063] Furthermore, the surfactant is any one selected from polyoxyethylene castor oil, porosium, sodium stearate, and Tween-80.

[0064] As a further advantageous effect of the above-mentioned configuration, the surfactant is employed to exert emulsifying and wetting action, to increase the solubility and stability of the active ingredient in the dosage form, and to enhance the penetration function. [Brief explanation of the drawings]

[0065] [Figure 1] 1 is a proton nuclear magnetic resonance spectrum (H1-NMR) of the first intermediate product prepared in Example 1 of the present invention. [Figure 2] 1 is a proton nuclear magnetic resonance spectrum of the second intermediate product prepared in Example 2 of the present invention. [Figure 3]1 is a proton nuclear magnetic resonance spectrum of the target product prepared in Example 3 of the present invention. [Figure 4] 1 is a proton nuclear magnetic resonance spectrum of the reference compound prepared in Example 4 of the present invention. [Figure 5] 1 is a proton nuclear magnetic resonance spectrum of the magnesium complex prepared in Example 5 of the present invention. [Figure 6] 1 is a proton nuclear magnetic resonance spectrum of the cadmium complex prepared in Example 5 of the present invention. [Figure 7] 1 shows the ultraviolet-visible absorption spectrum of the target product prepared in Example 3 in Experimental Example 6 of the present invention. [Figure 8] 1 shows the fluorescence emission spectrum of the target product prepared in Example 3 in Experimental Example 6 of the present invention. [Figure 9] FIG. 10 is a graph showing the change in the emission spectrum of dihydroethidium (DHE) depending on the light irradiation time in the presence of the target product prepared in Example 3 in Experimental Example 7 of the present invention. [Figure 10] 1 is a graph showing the change in the characteristic luminescence intensity of dihydroethidium (DHE) depending on the light irradiation time in Experimental Example 7 of the present invention. [Figure 11] FIG. 10 is a graph showing the change in the absorption spectrum of 1,3-diphenylbenzofuran (DPBF) depending on the light irradiation time in the presence of the target product prepared in Example 3 in Experimental Example 7 of the present invention. [Figure 12] 1 is a graph showing the decomposition rate of the target product 1,3-diphenylbenzofuran (DPBF) prepared in Example 3 in Experimental Example 7 of the present invention. [Figure 13] FIG. 10 is a graph showing the change in absorption spectra of the target product and 1,3-diphenylbenzofuran (DPBF) prepared in Example 3 with respect to the light irradiation time in Experimental Example 8 of the present invention. [Figure 14] FIG. 10 is a graph showing the change in the photodecomposition rate of the target product prepared in Example 3 depending on the power intensity in Experimental Example 8 of the present invention. [Figure 15] 1 is a high performance liquid chromatograph showing the photodecomposition process of the target product prepared and obtained in Example 3 in Experimental Example 8 of the present invention. [Figure 16] FIG. 10 is a diagram showing the detection of cellular phototoxicity of the target product prepared in Example 3 in Experimental Example 9 of the present invention. [Figure 17] FIG. 10 is a diagram showing the detection of cellular phototoxicity of the degradation products prepared and obtained in Example 3 in Experimental Example 9 of the present invention. [Figure 18] FIG. 10 is a diagram showing the fluorescent signal of a lysosomal probe in Experimental Example 10 of the present invention. [Figure 19] FIG. 10 is a diagram showing the fluorescence signal of the target product prepared in Example 3 in Experimental Example 10 of the present invention. [Figure 20] FIG. 10 shows the superimposed fluorescent signals of the lysosomal probe and the target product in Experimental Example 10 of the present invention. [Figure 21] This figure shows the biological fluorescence imaging of the target product prepared in Example 3 at a depth of 1 mm in Experimental Example 11 of the present invention. [Figure 22] This figure shows the in vivo fluorescence imaging of the target product prepared in Example 3 at a depth of 4 mm in Experimental Example 11 of the present invention. [Figure 23] This figure shows the biological fluorescence imaging of the target product prepared in Example 3 at a depth of 7 mm in Experimental Example 11 of the present invention. [Figure 24] This figure shows the biological fluorescence imaging of the target product prepared in Example 3 at a depth of 10 mm in Experimental Example 11 of the present invention. [Figure 25] FIG. 12 is a graph showing the changes in tumor volume in mice in the target product prepared in Example 3 and the light control group in Experimental Example 12 of the present invention. [Figure 26] FIG. 12 is a graph showing the weight changes of mice in the target product prepared in Example 3 and the light control group in Experimental Example 12 of the present invention. [Figure 27]FIG. 10 is a graph showing changes in tumor volume in mice for the degradation product group prepared and obtained in Example 3 in Experimental Example 12 of the present invention. [Figure 28] FIG. 10 is a graph showing changes in mouse body weight in the degradation product group prepared in Example 3 in Experimental Example 12 of the present invention. [Figure 29] 1 shows the ultraviolet-visible absorption spectrum of the target product prepared in Example 3 under continuous light irradiation in Comparative Example 1 of the present invention. [Figure 30] 1 shows the fluorescence emission spectrum of the target product prepared in Example 3 under continuous light irradiation in Comparative Example 1 of the present invention. [Figure 31] 1 shows the ultraviolet-visible absorption spectrum of the reference compound prepared in Example 4 under continuous light irradiation in Comparative Example 1 of the present invention. [Figure 32] 1 shows the fluorescence emission spectrum of the reference compound prepared in Example 4 under continuous light irradiation in Comparative Example 1 of the present invention. [Figure 33] FIG. 10 is a diagram showing cell imaging of the target product prepared in Example 3 at 0 minutes of light irradiation in Comparative Example 2 of the present invention. [Figure 34] FIG. 10 is a diagram showing cell imaging of the target product prepared in Example 3 after 5 minutes of light irradiation in Comparative Example 2 of the present invention. [Figure 35] FIG. 10 shows cell imaging of the target product prepared in Example 3 after 10 minutes of light irradiation in Comparative Example 2 of the present invention. [Figure 36] FIG. 10 is a diagram showing cell imaging at 0 minutes of light irradiation of the reference compound prepared in Example 4 in Comparative Example 2 of the present invention. [Figure 37] FIG. 10 is a diagram showing cell imaging of the reference compound prepared in Example 4 after 5 minutes of light irradiation in Comparative Example 2 of the present invention. [Figure 38] FIG. 10 shows cell imaging of the reference compound prepared in Example 4 after 10 minutes of light irradiation in Comparative Example 2 of the present invention. [Figure 39]FIG. 10 is a diagram showing biofluorescence imaging of the target product prepared in Example 3 at 0 minutes of light irradiation in Comparative Example 3 of the present invention. [Figure 40] FIG. 10 shows the in vivo fluorescence imaging of the target product prepared in Example 3 after 5 minutes of light irradiation in Comparative Example 3 of the present invention. [Figure 41] FIG. 10 shows the in vivo fluorescence imaging of the target product prepared in Example 3 after 10 minutes of light irradiation in Comparative Example 3 of the present invention. [Figure 42] FIG. 10 is a diagram showing in vivo fluorescence imaging of the reference compound prepared in Example 4 at 0 minutes of light irradiation in Comparative Example 3 of the present invention. [Figure 43] FIG. 10 is a diagram showing in vivo fluorescence imaging of the reference compound prepared in Example 4 after 5 minutes of light irradiation in Comparative Example 3 of the present invention. [Figure 44] FIG. 10 shows in vivo fluorescence imaging of the reference compound prepared in Example 4 after 10 minutes of light irradiation in Comparative Example 3 of the present invention. [Figure 45] 1 is a photograph of a mouse nail treated with the target product prepared in Example 3 in Comparative Example 4 of the present invention. [Figure 46] 10 is a photograph of the nails of a mouse in the light control group in Comparative Example 4 of the present invention. [Figure 47] 1 is a thermo-imaging photograph of a mouse treated with the target product prepared in Example 3 in Comparative Example 4 of the present invention. [Figure 48] 10 is a photograph of thermoimaging of mice in the light control group in Comparative Example 4 of the present invention. [Figure 49] 1 is a photograph of a mouse nail treated with the reference compound prepared in Example 4 in Comparative Example 4 of the present invention. [Figure 50] 1 is a thermo-imaging photograph of a mouse treated with the reference compound prepared in Example 4 in Comparative Example 4 of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0066] The principles and features of the present invention will be explained below with reference to the drawings, but the examples given here are merely for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention. [Example]

[0067] Example 1: Preparation of the first intermediate product [ka]

[0068] Tetrakis(pentafluorophenyl)porphodilactone (303 mg, 0.3 mmol) and potassium carbonate (1.24 g, 9 mmol) were added to a reaction vessel, and 7 mL of tetrahydrofuran and 1 mL of water were added to dissolve the mixture. The mixture was reacted at 80°C for 24 hours, after which the heating was stopped and the mixture was allowed to cool to room temperature. The reaction solvent was evaporated to dryness, and the mixture was dissolved in dichloromethane, filtered through diatomaceous earth (200 mesh), recrystallized with acetone / petroleum ether, filtered, and dried under vacuum (-0.1 MPa, 10 hours) to obtain 180 mg of the first intermediate product as a green solid.

[0069] The proton nuclear magnetic resonance spectrum of the first intermediate product is shown in Figure 1. The characterization data are as follows:

[0070] 1 H NMR (CDCl3,400MHz): δ9.62(d,J=4.8Hz,1H),9.39(d,J=2.8Hz,1H),8.69(d,J=4.8Hz,1H),8.53(d,J=3.6Hz,1H),-0.63(s,1H),-0.89(s,1H). 19FNMR(CDCl3,471MHz):δ-58.47(dd,J=19.7Hz,J=14.6Hz,1F),-59.39(d,J=6.6Hz,1F),-59.44(d,J=6.1Hz, 1F),-59.60(dd,J=19.7Hz,J=14.6Hz,1F),-61.20(d,J=6.6Hz,1F),-61.25(d,J=7.1Hz,1F),-72.48(t,J=21 0.2Hz, 1F), -73.61 (t, J = 20.1Hz, 1F), -75.90 (t, J = 20.2Hz, 1F), -77.55 (t, J = 20.2Hz, 1F), -82.01 (dd, J = 19.7Hz, J = 14.6Hz, 1F), -82.78 to -82.92 (m, 3F), -83.36 (t, J = 20.2Hz, 1F), -83.66 to -83.82 (m, 3F). HR-MS (ESI + ) m / z [M+H] + :Calculated value:C 41 H7F 18 N4O4961.0174, measured value: 961.0152.

[0071] Example 2: Preparation of the second intermediate product [ka]

[0072] The first intermediate product (94 mg, 0.10 mmol) and dimethylamine hydrochloride (1.6 g, 20 mmol) were added to 5 mL of N,N-dimethylformamide, and the mixture was refluxed for 36 h. After that, it was cooled to room temperature and precipitated by adding 50 mL of deionized water, filtered, and dried under vacuum (-0.1 MPa, 10 h). Under nitrogen gas protection, 80 mg of the above product and 0.5 mL of methyl trifluoromethanesulfonate were added to 5 mL of trimethyl phosphate, and the mixture was reacted at room temperature for 24 h. The precipitate was added to 50 mL of dichloromethane, centrifuged (4000 r, 10 min), and the solid was washed with 20 mL of dichloromethane and dried under vacuum (-0.1 MPa, 10 h). 100 mg of the second intermediate product, a purple solid, was obtained.

[0073] The proton nuclear magnetic resonance spectrum of the second intermediate product is shown in Figure 2. The characterization data were as follows:

[0074] 1 H NMR (CD3OD, 400MHz): δ9.74(d,J=5.6Hz,1H),9.45(d,J=3.6Hz,1H),9.13(d,J=5.2Hz,1H),8.83(d,J=4.8Hz,1H),4.19(d,J=4.0Hz,36H). 19 FNMR(CD3OD,471MHz):δ-135.25(t,J=15.0Hz,1F),-136.12(dd,J=26.3Hz,J=12 .2Hz,2F),-136.88(t,J=16.0Hz,1F),-137.56(d,J=11.8Hz,1F),-137.62(d,J= 13.1Hz,1F),-138.01(t,J=14.1Hz,1F),-138.31(d,J=15.5Hz,2F),-139.32(d, J=15.0Hz,2F),-140.62(t,J=14.6Hz,1F),-141.49(t,J=13.2Hz,1F).HR-MS(ESI + )m / z[M]4 + :Calculated value:C 53 H 42 F 14 N8O4 4+ 280.0771, Actual value: 280.0767.

[0075] Example 3: Preparation of target product [ka]

[0076] The second intermediate product (110 mg, 0.1 mmol) and zinc acetate (200 mg, 1 mmol) were dissolved in 20 mL of methanol. The mixture was stirred overnight at room temperature and evaporated to dryness. The mixture was dissolved in 5 mL of acetonitrile and centrifuged (4000 r, 5 min). The supernatant was collected and precipitated with 50 mL of dichloromethane. The precipitate was centrifuged (4000 r, 10 min), and dried under vacuum (-0.1 MPa, 10 h) to obtain 110 mg of the target product—a degradable near-infrared photosensitizer molecule—as a green solid.

[0077] The proton nuclear magnetic resonance spectrum of the target product is shown in Figure 3. The characterization data are as follows:

[0078] 1 H NMR (CD3OD, 400MHz): δ9.51(d,J=5.2Hz,1H),9.32(d,J=4.4Hz,1H),8.85(d,J=4.8Hz,1H),8.65(d,J=4.4Hz,1H),4.18(s,36H). 19 FNMR(CD3OD,471MHz):δ-136.39(dd,J=27.7Hz,J=12.7Hz,2F),-136.71(t,J= 14.6Hz,1F),-137.13(t,J=14.6Hz,1F),-137.95(dd,J=28.2Hz,J=13.2Hz,2F ),-138.78(d,J=15.5Hz,2F),-138.90(s,1F),-138.99(s,1F),-140.03(d,J= 16.0Hz,2F),-141.28(t,J=13.2Hz,1F),-142.04(t,J=12.7Hz,1F).HR-MS(ESI + )m / z[M]4 + :Calculated value:C 53 H 40 F 14 N8O4Zn 4+ 295.5555, Actual value: 295.5557.

[0079] Example 4: Preparation of Reference Compounds A reference compound—a non-degradable photosensitizer was prepared in the same manner as in Examples 1 to 3, except that tetrakis(pentafluorophenyl)porpholactone (298 mg, 0.3 mmol) was used as the starting reactant. [ka]

[0080] The proton nuclear magnetic resonance spectrum of the reference compound is shown in Figure 4. The characterization data were as follows:

[0081] 1 H NMR (CD3OD, 400MHz): δ9.21(d,J=4.4Hz,2H),8.64(d,J=4.4Hz,2H),8.24(s,2H),4.16(d,J=9.2Hz,36H). 19 FNMR(CD3OD,471MHz):δ?137.15(dd,J=34.8Hz,J=16.9Hz,4F),-138.82(t,J=20.7Hz, 2F),-139.49(d,J=20.2Hz,4F),-139.64(s,2F),-142.36(t,J=14.6Hz,2F).HR-MS(ESI + )m / z[M]4 + :Calculated value:C 54 H 42 F 14 N8O2Zn 4+ 291.0619, Actual value: 291.0620.

[0082] Example 5: Preparation of complexes with different central metals [ka]

[0083] The first intermediate product (110 mg, 0.1 mmol) and magnesium acetate / cadmium acetate / copper acetate / manganese acetate (10 eq.) were dissolved in 10 mL of N,N-dimethylformamide. The mixture was reacted at 150 °C for 8 h, then the heating was stopped and the mixture was allowed to cool to room temperature. The reaction solvent was evaporated to dryness, and the mixture was dissolved in dichloromethane, filtered through diatomaceous earth (200 mesh), and recrystallized from acetone / petroleum ether to obtain complexes with magnesium, cadmium, copper, and manganese as the central metals, respectively.

[0084] The proton nuclear magnetic resonance spectrum of the magnesium complex is shown in Figure 5. The characterization data were as follows:

[0085] 1 H NMR(CDCl3,400MHz):δ9.47(d,J=4.8Hz,1H),9.29(d,J=4.4Hz,1H),8.48(d,J=4.8Hz,1H),8.38(d,J=4.4Hz,1H).HR-MS(ESI + ) m / z [M+H] + :Calculated value:C 41 H5F 18 MgN4O4982.9868, Measured value: 982.9887.

[0086] The proton nuclear magnetic resonance spectrum of the cadmium complex is shown in Figure 6. The characterization data were as follows:

[0087] 1 H NMR(CDCl3,400MHz):δ9.61(d,J=4.4Hz,1H),9.48(d,J=4.0Hz,1H),8.67(d,J=4.8Hz,1H),8.60(d,J=4.4Hz,1H).HR-MS(ESI + ) m / z [M+H] + :Calculated value:C 41 H5CdF 18 N4O41072.9063, Measured value: 1072.9077.

[0088] Copper complex: HR-MS (ESI + ) m / z [M+H]+ :Calculated value:C 41 H5CuF 18 N4O41021.9314, Measured value: 1021.9299.

[0089] Manganese complex: HR-MS (ESI + ) m / z [M+H] + :Calculated value:C 41 H5F 18 MnN4O41013.9398, Measured value: 1013.9410.

[0090] Experimental Example 6: Detection of photophysical properties The target product prepared in Example 3 was scanned for UV-visible absorption and emission spectra, as shown in Figures 7 and 8. Figure 7 reveals that the absorption spectrum of the target product covers the visible and near-infrared regions, with strong absorption in the red-near-infrared region of 600 nm to 750 nm. Figure 8 reveals that the fluorescence spectrum of the target product, obtained by photoexcitation, is also located in the near-infrared region. This indicates that the target product has deep tissue penetration depth for photodynamic therapy or biofluorescence imaging.

[0091] Experimental Example 7: Detection of reactive oxygen species In an air atmosphere, dihydroethidium (DHE) was added as a superoxide anion radical probe to an aqueous solution (10 μM) of the target product prepared in Example 3, and the light irradiation conditions were a 700 nm light-emitting diode lamp, 1.25 mW / cm. 2 When the dihydroethidium was oxidized by superoxide radicals, the fluorescence of oxygenated ethidium was detected, and the fluorescence intensity increased with the extension of the light irradiation time. As a result, as shown in Figures 9 and 10, it is suggested that the target product has the ability to generate superoxide anion radicals under light irradiation.

[0092] The phosphorescence of singlet oxygen is easily quenched by water molecules, and the singlet oxygen generated from the target product was measured spectrophotometrically by chemical capture. 1,3-Diphenylbenzofuran (DPBF) is the most common singlet oxygen scavenger, and it can rapidly react with singlet oxygen to produce a colorless substance. The change in absorbance at its characteristic absorption peak (416 nm) in the UV-visible absorption spectrum reflects the singlet oxygen content. The light irradiation conditions were a 700 nm light-emitting diode lamp, 1.25 mW / cm. 2 When the 416 nm characteristic absorption of 1,3-diphenylbenzofuran (DPBF) was measured, it gradually weakened, suggesting that the target product had the ability to generate singlet oxygen under light irradiation, as shown in Figures 11 and 12.

[0093] Experimental Example 8: Monitoring autolysis In the test of Experimental Example 7, the light irradiation conditions were a 700 nm light emitting diode lamp, 1.25 mW / cm 2 From 0 to 2 min, the characteristic absorption of 1,3-diphenylbenzofuran (DPBF) at 416 nm gradually weakened. By increasing the light irradiation intensity and extending the irradiation time, the light irradiation conditions were changed to a 700 nm light-emitting diode lamp, 10 mW / cm. 2 When the irradiation time was 2 to 4 min, the characteristic absorption of the target product at 440 nm and 710 nm decreased significantly, suggesting that the target product was decomposed by light irradiation, as shown in Figure 13. Furthermore, as shown in Figure 14, the decomposition rate became significantly faster with increasing light irradiation intensity.

[0094] The photodegradation process was further monitored by HPLC, and the light irradiation conditions were a 700 nm light-emitting diode lamp, 2.5 mW / cm 2 When the irradiation time was 0 to 10 min, the characteristic peak of the target product gradually decreased with the increase in irradiation time, and a new characteristic peak (decomposition product) generated at 5 min gradually increased. As a result of collecting the decomposition product, as shown in Figure 15, it was suggested that the target product was autolyzed under light irradiation and converted into decomposition products.

[0095] Experimental Example 9: Cellular phototoxicity The cells used in this experiment were HeLa human cervical cancer cells. The cells were cultured in DMEM complete medium (Genyo, 500 mL) supplemented with 10% inactivated fetal bovine serum (Sekika, 500 mL) and 1% penicillin-streptomycin (Corning, 100 mL) at 37°C in a 5% carbon dioxide atmosphere.

[0096] Subcultured HeLa cells were digested with trypsin and then dispersed in medium at an appropriate concentration. The dispersed HeLa cells were seeded into a flat-bottom 96-well plate, with 200 μL of medium per well, and approximately 10 cells per well. One group of medium without cells was used as a blank control. After culturing the cells in the dark for 24 h, the medium was removed, and either 100 μL of fresh medium and 100 μL of a previously prepared mixture of the target product, or 100 μL of fresh medium and 100 μL of a mixture of the degradation products prepared in Example 3, was added. The sample was diluted to a concentration gradient of 0–4 μM. After culturing for 24 h in the dark, the medium was removed, and each well was washed three times with PBS buffer (pH 7.4). 100 μL of PBS buffer was added to each well, and the light irradiation conditions were a 700 nm light-emitting diode lamp at 2.5 mW / cm. 2The incubation was continued for 10 min. The PBS buffer was removed from each well, and 200 μL of fresh medium was replaced. The incubation was continued for 24 h. After incubation, the medium was removed, and each well was washed three times with PBS buffer. Then, 100 μL of 10% CCK-8 reagent (Cell Counting Kit-8) prepared with medium was added to each well and incubated for 2 h. During this process, 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt (WST-8) in the CCK-8 reagent was reduced by viable cells with the aid of an electron-binding reagent to form a water-soluble yellow formazan product, which changed the absorbance of the solution at 450 nm. This change was directly proportional to the number of viable cells. The absorbance change at 450 nm in each well was measured using an enzyme-linked immunosorbent assay, and the cell viability at each incubation concentration was calculated using the following formula:

[0097] CV = (As-Ab) / (Ac-Ab) × 100%.

[0098] Here, CV refers to cell viability, and As, Ac, and Ab refer to the absorbance of cells in the incubating compound group, the absorbance of cells in the blank group, and the absorbance of the blank control group, respectively.

[0099] The median lethal concentration (IC50) of the target product in HeLa cells was calculated from the cell viability at each incubation concentration and was found to be 1.0 μM. Figure 16 shows the cellular phototoxicity of the target product, which exhibits strong phototoxicity under light irradiation. Figure 17 shows the cellular phototoxicity of the degradation products, which exhibit no phototoxicity.

[0100] Experimental Example 10: Cellular Fluorescence Imaging Subcultured HeLa cells were digested with trypsin and dispersed in medium at the appropriate concentration. Dispersed HeLa cells were seeded into confocal dishes and cultured for 24 h. After 24 h, 100 μL of a pre-prepared target product medium solution was added and diluted to a 10 μM sample concentration. After 12 h of incubation, a lysosomal green fluorescent probe was added. After 15 min of co-incubation, the medium was removed and the cells were washed three times with PBS buffer (pH 7.4). Fluorescence imaging was performed using an ISS-integrated laser scanning confocal fluorescence lifetime imaging system. The target product fluorescence signal was excited at 700 nm using a supercontinuous laser and collected through a 715 nm high-pass filter. The lysosomal probe fluorescence signal was excited at 488 nm using a laser instrument and collected through a 525 / 50 nm band-pass filter.

[0101] As shown in FIG. 18, in the diagram showing the fluorescent signal of the lysosomal probe, the lysosomal probe showed a strong fluorescent signal in the green channel.

[0102] As shown in FIG. 19, in the diagram showing the fluorescent signal of the target product, the target product exhibited a strong fluorescent signal in the near infrared region.

[0103] As shown in Figure 20, in the diagram showing the superimposed fluorescence signals of the lysosomal probe and the target product, the near-infrared fluorescence of the target product and the green fluorescence of the lysosomal probe were well superimposed, suggesting that the target product has potential as a near-infrared live-cell fluorescence imaging probe and is located in lysosomes.

[0104] Experimental Example 11: Depth of In vivo Fluorescence Imaging All animal experiments in vivo were conducted in strict compliance with China's Animal Experimentation Practice Regulations. Five-week-old, male BALB / C nude mice weighing 16-25 g were used. Mice were injected subcutaneously in the abdomen with 100 μL of the target product in PBS (0.2 mg / mL) at a dose of 0.2 mg / kg. Thirty minutes after administration, the mice were placed in the imaging device and anesthetized in a 2 L / min oxygen / 2% isoflurane gas mixture. The excitation wavelength for the target product was 710 nm, and the image acquisition wavelength was 750 nm. A layer of mouse skin (1 mm per layer) was placed over the injection site, and changes in the intensity of the fluorescent signal were monitored until the signal disappeared.

[0105] As shown in FIG. 21, in the image showing the in vivo fluorescence imaging of the target product at a depth of 1 mm, the target product showed a strong fluorescent signal subcutaneously.

[0106] As shown in FIG. 22, in the diagram showing the biological fluorescence imaging of the target product at a depth of 4 mm, the detected fluorescence signal of the target product is somewhat weak.

[0107] As shown in FIG. 23, in the diagram representing the biological fluorescence imaging of the target product at a depth of 7 mm, the detected fluorescence signal of the target product becomes weaker as the imaging depth increases.

[0108] As shown in FIG. 24, in the diagram showing the biological fluorescence imaging of the target product at a depth of 10 mm, the fluorescence signal of the target product suggests that the fluorescence signal can be effectively detected even at a depth of 10 mm.

[0109] Experimental Example 12: Example of drug phototherapy at the animal level All animal experiments in vivo were conducted in strict compliance with China's animal experiment implementation regulations. Five-week-old, male BALB / C nude mice weighing 16–25 g were used. Each mouse was subcutaneously injected with 100 μL (5 × 10) of HeLa human cervical cancer cells in the right hind leg. 6 ) were inoculated to establish tumor models, and experiments were performed two weeks later.

[0110] The mice were randomly assigned to groups by number, and the experiments were conducted with six mice per group divided into a light control group (light irradiation), a dark control group (target product, degradation product), and an experimental group (target product + light irradiation, degradation product + light irradiation). The dark control group and the experimental group were administered by in situ injection, respectively. The dose was 0.2 mg / kg, and 20 μL (0.2 mg / mL) of the target product or degradation product in PBS solution was administered by in situ injection. 30 min after administration, the light control group and the experimental group underwent irradiation treatment, and the light irradiation conditions were a 700 nm light-emitting diode lamp, 100 mW / cm. 2 Each mouse was exposed to light for 10 minutes. After treatment, the mice were kept in cages without any light shielding, and tumor volume was measured with a caliper every two days, and the mice were weighed. Treatment was continued for two weeks.

[0111] As shown in Figure 25, which shows the changes in tumor volume in mice in the target product and light control groups, the target product can effectively suppress the growth of subcutaneous tumors under single light irradiation conditions, but the tumor growth in mice in the light control group that was not administered the target product and the dark control group that was administered the target product was rapid.

[0112] As shown in FIG. 26, in the graph showing the weight changes of mice in the target product and light control groups, the weights of mice in both the experimental group and the control group did not obviously decrease.

[0113] As shown in Figure 27, in the graph showing the tumor volume changes in mice in the decomposition product group, the decomposition products did not inhibit tumor growth under light irradiation and dark control conditions, suggesting that the decomposition products are not photoactive and do not damage cells or tissues.

[0114] As shown in Figure 28, in the graph showing the weight changes of mice in the decomposition product group, the weight of mice in the irradiation group of the decomposition product group and the dark control group did not obviously decrease, suggesting that the decomposition product has no obvious side effects on the living body.

[0115] Comparative Example 1: Example of the photodecomposition process of a photosensitizer in solution The target product obtained in Example 3 was placed under a light-emitting diode lamp and irradiated under the light irradiation conditions of 700 nm and 10 mW / cm 2 The UV-visible absorption and emission spectra of the target product were recorded during the light irradiation process. As shown in Figure 29, the characteristic absorption of the target product at 440 nm and 710 nm decreased significantly, and as shown in Figure 30, the characteristic emission of the target product at 740 nm also gradually weakened. These results suggest that the target product was decomposed by light irradiation.

[0116] As a control, the reference compound prepared in Example 4 was placed under a light-emitting diode lamp and irradiated under the conditions of 640 nm, 10 mW / cm 2 During the irradiation process, the UV-visible absorption and emission spectra of the reference compound were recorded.

[0117] As shown in FIG. 31, in the ultraviolet-visible absorption spectrum of the reference compound under continuous light irradiation, no change was observed in the absorption spectrum of the reference compound under continuous light irradiation conditions.

[0118] As shown in Figure 32, in the fluorescence emission spectrum of the reference compound under continuous light irradiation, no obvious change was observed in the fluorescence emission intensity of the reference compound, suggesting that the reference compound does not decompose under light irradiation conditions.

[0119] Comparative Example 2: Example of the photodecomposition process of a photosensitizer in cells HeLa cells were seeded in a confocal dish and cultured for 24 hours. After incubation, 100 μL of a pre-prepared medium solution of the target product or reference compound was added and diluted to a concentration of 10 μM. After 12 hours of incubation, the samples were washed three times with PBS buffer (pH 7.4). Fluorescence imaging was performed using an ISS-integrated laser scanning confocal fluorescence lifetime imaging system. The target product fluorescence signal was collected using a 715 nm high-pass filter, and the reference compound fluorescence signal was collected using a 670 / 50 nm band-pass filter.

[0120] As shown in Figure 33, in the cell imaging at 0 min of light irradiation of the target product, the target product has a strong fluorescent signal in the near-infrared region. The dish was placed under a light-emitting diode lamp and irradiated with light under the conditions of 700 nm and 10 mW / cm. 2 Imaging was performed once every 5 min.

[0121] As shown in FIG. 34, in the image showing cell imaging of the target product after 5 minutes of light irradiation, the fluorescent signal of the target product was clearly weakened 5 minutes after light irradiation.

[0122] As shown in Figure 35, in the cell imaging of the target product after 10 minutes of light irradiation, the fluorescence signal of the target product was almost quenched 10 minutes after light irradiation, suggesting that the target product in the cells was autolyzed under light irradiation.

[0123] As a control, in the image showing cell imaging at 0 min of light irradiation with the reference compound, as shown in Figure 36, the reference compound showed a strong fluorescent signal. The dish was placed under a light-emitting diode lamp and irradiated with light at 640 nm and 10 mW / cm. 2 Imaging was performed once every 5 min.

[0124] As shown in FIG. 37, in the image showing cell imaging after 5 minutes of light irradiation with the reference compound, the fluorescent signal of the reference compound does not weaken 5 minutes after light irradiation.

[0125] As shown in FIG. 38, in the diagram showing cell imaging after 10 minutes of light irradiation with the reference compound, no clear change was observed in the intensity of the fluorescent signal of the reference compound 10 minutes after light irradiation.

[0126] Comparative Example 3: Example of the photodegradation process of photosensitizers at the animal level All animal experiments in vivo were conducted in strict compliance with China's animal experiment implementation regulations. Five-week-old, male BALB / C nude mice weighing 16–25 g were used. Each mouse was subcutaneously injected with 100 μL (5 × 10) of HeLa human cervical cancer cells in the right hind leg. 6 ) were inoculated to establish tumor models, and experiments were performed two weeks later.

[0127] 20 μL (0.2 mg / mL) of the target product or reference compound in PBS was administered into the mouse tumor via in situ injection at a dose of 0.2 mg / kg. 30 min after administration, the mouse was placed in the imaging device and anesthetized in a 2 L / min oxygen / 2% isoflurane gas mixture. The excitation wavelength for the target product was 710 nm, and the imaging wavelength was 750 nm. The excitation wavelength for the reference compound was 650 nm, and the imaging wavelength was 690 nm.

[0128] As shown in Figure 39, in the diagram showing the in vivo fluorescence imaging of the target product at 0 min of light irradiation, the target product showed a strong fluorescent signal in the mouse tumor. The tumor area of ​​the mouse was irradiated under a light-emitting diode lamp, and the light irradiation conditions were 700 nm, 100 mW / cm. 2 Imaging was performed once every 5 min.

[0129] As shown in Figure 40, in the image showing the in vivo fluorescence imaging of the target product 5 minutes after light irradiation, the fluorescent signal in the mouse tumor was obviously weaker 5 minutes after light irradiation, suggesting that the target product was autolyzed during the light irradiation treatment.

[0130] As shown in Figure 41, in vivo fluorescence imaging of the target product at 10 minutes after irradiation, the fluorescent signal in the mouse tumor was completely quenched 10 minutes after irradiation, and the target product was completely converted to a degradation product that was not photoactive upon completion of treatment.

[0131] As a control, in the diagram showing the in vivo fluorescence imaging of the reference compound at 0 min of light irradiation, the reference compound showed a strong fluorescent signal in the mouse tumor, as shown in Figure 42. The tumor area of ​​the mouse was irradiated under a light-emitting diode lamp, and the light irradiation conditions were 640 nm, 100 mW / cm. 2 Imaging was performed once every 5 min.

[0132] As shown in FIG. 43, in the diagram showing the in vivo fluorescence imaging of the reference compound after 5 minutes of light irradiation, the fluorescence signal in the mouse tumor did not weaken 5 minutes after light irradiation.

[0133] As shown in Figure 44, in the diagram showing the in vivo fluorescence imaging of the reference compound after 10 minutes of light irradiation, no obvious change was observed in the intensity of the fluorescent signal in the mouse tumor 10 minutes after light irradiation, suggesting that the reference compound does not have photodegradation ability.

[0134] Comparative Example 4: Example of photosensitization side reactions of photosensitizers at the animal level All animal experiments in vivo were conducted in strict compliance with China's Animal Experiment Implementation Regulations. Five-week-old, male, BALB / C nude mice weighing 16-25 g were used. Mice were randomly assigned to groups based on their numbers, and experiments were conducted with three mice per group: target product group, reference compound group, and light control group. Mice were injected subcutaneously into the abdomen with 20 μL (0.2 mg / mL) of the target product, reference compound solution, or PBS solution at a dose of 0.2 mg / kg. After injection, the injection site was irradiated with light. The target product was irradiated with a 700 nm light-emitting diode lamp, and the reference compound was irradiated with light at 100 mW / cm. 2 The mice were exposed to simulated natural light at 10 mW / cm². The light control group was left untreated. The mice were observed and their body temperatures were recorded using a thermo-imaging device. The mice in the three groups were then exposed to simulated natural light at 10 mW / cm². 2 Then, it was irradiated with light for 1 hour.

[0135] The mice in the target product group and the light control group showed no abnormal reactions when exposed to natural light. As shown in Figures 45 and 46, the limbs of the mice were strong and the condition of the nails was normal. As shown in Figures 47 and 48, there was no obvious change in body temperature.

[0136] When exposed to natural light, the mice in the reference compound group showed obvious photosensitization side effects, including a decrease in motor ability, muscle weakness, and obvious curling of nails, as shown in Figure 49. As shown in Figure 50, the mice's body temperature also showed a clear drop.

[0137] Solvates of porphyrin compounds in water / ethanol / acetonitrile, noncovalent complexes with proteins, and prodrugs were examined in experiments similar to those in Examples 6 to 12, and it was found that these substances also had effects similar to those of the compound in Example 3. For compounds with substituents different from those in Example 3, the influence of the substituents on the peripheral benzene rings on the electronic structure of the macrocycle itself was negligible, and the photosensitizer molecules reacted with the reactive oxygen species generated by photoactivation during the decomposition process depending on the lactone units in the structure. As a result of examination in experiments similar to those in Examples 6 to 12, it was found that these substances also had effects similar to those of the compound in Example 3.

[0138] In conclusion, the decomposable near-infrared photosensitizer of the present invention has an absorption spectrum in the near-infrared region and a large extinction coefficient, and compared with reference compounds, it generates reactive oxygen species upon irradiation with light and is decomposed, and after irradiation is terminated, it is completely converted into a non-photoactive, low-toxic substance. Therefore, the decomposable near-infrared photosensitizer of the present invention exhibits high phototoxicity at both the cellular and biological levels, and its strong fluorescence can be used for fluorescent labeling to monitor its own decomposition in real time.

[0139] The above content is merely a preferred embodiment of the present invention, and does not limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should all be included in the protection scope of the present invention.

Claims

1. A degradable near-infrared photosensitizer comprising, as an active ingredient, any one selected from the group consisting of a porphyrin compound represented by formula 1, and its pharmaceutically acceptable salts, solvates, and non-covalent complexes with proteins; 【Chemistry 1】 During the ceremony, 【Chemistry 2】 teeth, 【Transformation 3】 and Ar １ and Ar ２ teeth, 【Chemistry 4】 and R １ , R ３ , R ４ , R ５ , R ７ , R ８ , R 9 , R 10 , R 12 , and R 13 are F; R 2 , R 6 , and R 11 are —N + (CH 3 ) 3 ; M is Zn.

2. A method for preparing the decomposable near-infrared photosensitizer of claim 1, comprising the following steps 1 to 3: In step 1 of preparing the first intermediate product, 100 mg of porphodilactone and an alkali equivalent to 30 to 50 times the amount of the porphodilactone are dissolved in 5 mL to 20 mL of a first solvent in a hydrothermal reaction vessel, and the mixture is reacted at 80°C to 200°C for 24 hours to 72 hours to obtain the first intermediate product, the reaction formula of which is as follows: 【Transformation 6】 In the second step of preparing the second intermediate product, 100 mg of the first intermediate product obtained in the first step is subjected to a water-soluble modification reaction in 5 mL to 50 mL of a second solvent to obtain a second intermediate product, the reaction formula of which is as follows: 【Transformation 7】 In step 3 for preparing the target product, 100 mg of the second intermediate product obtained in step 2 and a zinc salt equivalent to 10 times the amount of the second intermediate product are dissolved in 20 to 100 mL of a third solvent, and reacted at 20 to 80°C for 2 to 16 hours to obtain the target product shown in formula 1, and the reaction equation is as follows: 【Transformation 8】 A method for preparing a decomposable near-infrared photosensitizer, comprising:

3. In step 1, the alkali is any one or a mixture of two or more selected from sodium hydroxide, potassium hydroxide, potassium carbonate, sodium carbonate, triethylamine, N,N-diisopropylethylamine, sodium methoxide, and potassium tert-butoxide; the first solvent is any one or a mixture of two or more selected from water, methanol, ethanol, acetonitrile, acetone, dimethyl sulfoxide, N,N-dimethylformamide, and tetrahydrofuran; The method for preparing a decomposable near-infrared photosensitizer according to claim 2, characterized in that the reaction temperature is 100°C to 150°C and the reaction time is 48h to 72h.

4. 3. The method for preparing a decomposable near-infrared photosensitizer according to claim 2, wherein the second solvent is any one or a mixture of two or more selected from the group consisting of dimethyl sulfoxide, N,N-dimethylformamide, tetrahydrofuran, ethyl acetate, trimethyl phosphate, triethyl phosphate, and hexanol.

5. the third solvent is any one or a mixture of two or more selected from water, methanol, ethanol, acetonitrile, acetone, dimethyl sulfoxide, N,N-dimethylformamide, tetrahydrofuran, chloroform, and dichloromethane; The method for preparing the decomposable near-infrared photosensitizer according to claim 2, characterized in that the reaction temperature is 20°C to 60°C, and the reaction time is 3h to 5h.

6. 10. Use of the decomposable near-infrared photosensitizer of claim 1 as an active ingredient in the preparation of a photodynamic therapeutic drug.

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