Ultrasound Sensitizers

Abstract

To provide a compound that has reduced burden on patients and is applicable to SDT (Sonodynamic Therapy).SOLUTION: The present invention relates to an electron-withdrawing group carrying phenyl chlorin derivative or ethylene glycol derivative represented by formula (I) or a metal complex thereof or a pharmacologically acceptable salt thereof [R is -N(CH2COONa)2, aspartate residue or glutamic acid residue, or -NH(CH2CH2)nOH (n is an integer of 1 to 3)].SELECTED DRAWING: Figure 14

Description

[Technical field]

[0001] The present invention relates to a novel phenylchlorin derivative or ethylene glycol derivative bearing an electron-withdrawing group, or a pharmacologically acceptable salt thereof, and to a cancer treatment agent, a skin treatment agent, and an antibacterial agent containing the same as an active ingredient for use in sonodynamic therapy (SDT). The present invention also relates to novel phenylchlorin metal complex derivatives or ethylene glycol metal complex derivatives bearing an electron-withdrawing group, or pharmacologically acceptable salts thereof, which have been complexed with Cu or Mn metal to further increase their water solubility. [Background technology]

[0002] Sonodynamic therapy (SDT) is being used as a new cancer treatment. This is a treatment in which a certain type of porphyrin derivative is administered by intravenous injection or other methods, and selectively accumulates in cancer (tumor) tissue. Then, the cancer tissue is selectively destroyed by irradiating it with ultrasound, eliminating the cancer cells. This treatment makes use of two properties of porphyrin derivatives: their ability to selectively accumulate in cancer tissue and their ultrasound sensitization effect.

[0003] On the other hand, photophysicochemical diagnosis and therapy (PDT: Photodynamic Therapy) is a treatment method that uses light irradiation and takes advantage of two properties of porphyrin derivatives: their selective accumulation in cancer tissue and their photosensitizing effect.

[0004] One of the present inventors has been actively researching porphyrin derivatives that can be used in PDT, and has provided many derivatives including ATX-S10 (Patent Document 1).

[0005] On the other hand, as SDT sensitizers, 5-aminolevulinic acid hydrochloride (5-ALA) and chlorine Ce have been used for cancer diseases. 6 -Sn complex (T-Ce 6) and ethylene glycol-supported chlorin Sn complex (ACT4211) have been developed (Patent Document 2, Non-Patent Documents 1 and 2).

[0006] In recent years, it has become clear that PDT using 5-ALA is effective for the diagnosis and treatment of cancer. Attempts have also been made to treat cancer through SDT using 5-ALA. However, 5-ALA is a biosynthetic precursor of protoporphyrin IX, and due to the compound properties of protoporphyrin IX, namely low neovascular accumulation, a longest wavelength absorption edge of 630 nm, phototoxicity, and weak ultrasonic activation, it cannot be said to be a good SDT therapeutic agent.

[0007] On the other hand, T-Ce 6 and ACT4211 are chlorophyll derivatives derived from plants, and our long-term research has shown that their safety is not necessarily good because they are derived from plants, not animals. That is, the electron-withdrawing group-bearing phenylchlorin derivatives or ethylene glycol derivatives developed by the present inventors are derivatives obtained by synthesis through 4 to 5 steps from blood-derived protoporphyrin. The chlorins created by the present inventors are decomposed in vivo, metabolized by protoporphyrin, and excreted by the normal metabolic route, suggesting that they are particularly safe.

[0008] The applicants have filed a patent application for the precursor substance, a chlorin Mn metal complex (Patent Document 3), which is an ultrasonic sensitizer, as a compound for SDT. Meanwhile, the chlorin substances proposed thus far (Patent No. 5179245: Patent Document 4, Patent No. 5651426: Patent Document 5) were compounds whose phototoxicity could be avoided by forming a Mn complex.

[0009] We previously developed the aforementioned substance (Patent Document 3: JP 2018-199649 A), but the invented substance contained iodine molecules in its molecule, which caused some problems. Iodine molecules have the advantage of being used as a radiodiagnostic agent, but on the other hand, iodine molecules are thought to cause allergies, so there was a demand for compounds that do not contain iodine molecules in their molecules.

[0010] On the other hand, previous research by one of the inventors has revealed that phototoxicity can be eliminated by attaching an electron-withdrawing group to the porphyrin skeleton and forming a Mn complex. Furthermore, since chloral hydrate is sensitive to ultrasound, it was also known that the presence of two or more chlorine molecules was sufficient. Therefore, porphyrins with chlorinated phenyl groups were developed (Non-Patent Document 3), but they had very poor water solubility and could not be used as SDT sensitizers.

[0011] Therefore, we have been conducting research and development to ensure water solubility by combining the electron-withdrawing group-bearing phenylchlorin compound with a functional group having a polyvalent carboxylic acid group or a polyvalent hydroxyl group, such as aspartic acid (Patent Document 1), which has been shown to have a high affinity for cancer in previous studies. We have also investigated Mn, Cu and Fe complexes to further improve water solubility.

[0012] As a result of the research, they discovered derivatives that combine water solubility and cancer affinity by combining amino acids with two or more carboxylic acids or two or more ethylene glycol units.

[0013] The present invention has been completed based on the confirmation that the novel phenylphenylchlorin derivatives or ethylene glycol derivatives carrying electron-withdrawing groups are effective not only as cancer therapeutic agents but also as external preparations for SDT such as ointments and lotions for the treatment of infectious diseases.

[0014] To further improve water solubility, the novel chlorin derivatives can be converted into Mn, Fe, and Cu metal complexes. [Prior art documents] [Patent documents]

[0015] [Patent Document 1] Patent No. 3613599 [Patent Document 2] U.S. Patent No. 6462192 [Patent Document 3] JP 2018-199649 A [Patent Document 4] Patent No. 5179245 [Patent Document 5] Patent No. 5651426 [Non-patent literature]

[0016] [Non-Patent Document 1] Anticancer Res., 31(2) 501 (2011) [Non-Patent Document 2] Intergr. Cancer The., 7(2) 96 (2008) [Non-Patent Document 3] Cancer Sci., 98: 916 (2007) Summary of the Invention [Problem to be solved by the invention]

[0017] Therefore, an object of the present invention is to provide a phenylchlorin amino acid derivative or an ethylene glycol derivative carrying an electron-withdrawing group that can be used in sonodynamic therapy (SDT), and further to provide a cancer injection or topical agent for SDT, in particular a cancer therapeutic agent, a skin disease therapeutic agent, and an antibacterial agent, which contain, as an active ingredient, a Mn, Cu, or Fe complex of the phenylchlorin derivative or ethylene glycol derivative carrying an electron-withdrawing group. [Means for solving the problem]

[0018] In order to solve the above problems, the present invention provides, in one basic embodiment, a compound represented by the following formula (I):

[0019] [ka]

[0020] [In the formula, X is represented by the following formula:

[0021] [ka]

[0022] (The above substituent will be referred to as [Chemical Formula 2] below.) Or the following formula

[0023] [ka]

[0024] (The above substituent will be referred to as [Chemical formula 3] below.)

[0025] (wherein Y represents an electron-withdrawing group and is a halogen atom, NH2, OH, or OCH3), R is -N(CH2COONa)2 [which represents the residue of iminodiacetic acid], an aspartic acid residue or a glutamic acid residue, or -NH(CH2CH2O) n OH (n represents an integer from 1 to 3), M represents Mn, Fe or Cu; Z represents a ligand and is OAc or Cl. or a metal complex thereof, or a pharmacologically acceptable salt thereof.

[0026] In a more specific embodiment, the present invention relates to a compound represented by formula (I)-A type:

[0027] [ka]

[0028] And the following formula (I)-B type:

[0029] [ka]

[0030] (In the above formulas, X, R, Y, M, Z and n are the same as defined above.) or a metal complex thereof, or a pharmacologically acceptable salt thereof.

[0031] In another embodiment, the present invention relates to a cancer therapeutic agent, a skin disease therapeutic agent, or an antibacterial agent for SDT, which contains the above-mentioned electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative, or a metal complex or a pharmacologically acceptable salt thereof as an active ingredient, and in the case of a Mn complex, it can also be used as an MRI contrast agent.

[0032] More specifically, the present invention relates to a cancer therapeutic agent, skin disease therapeutic agent, or antibacterial agent for the above-mentioned SDT in the form of an injection, ointment, sublingual agent, or lotion, and can also be used as an MRI contrast agent.

[0033] That is, the basic aspect of the present invention is characterized in that it uses an electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative represented by the above formula (I), particularly an (I)-A type or (I)-B type phenylchlorin derivative or ethylene glycol derivative bearing an electron-withdrawing group, or a metal complex thereof, or a pharmacologically acceptable salt thereof, to treat cancer diseases, skin diseases, and infectious diseases by SDT. Effect of the Invention

[0034] The electron-withdrawing group-bearing phenylchlorin derivatives or ethylene glycol derivatives, or metal complexes or pharmacologically acceptable salts thereof provided by the present invention are water-soluble chlorin derivatives, and drugs containing these chlorin derivatives, such as injections, liquids, ointments, sublingual preparations, suppositories, and lotions, have good accumulation and permeability to affected areas, are non-phototoxic, and are sensitive to ultrasound, so that they do not impose a burden on patients in SDT therapy for the treatment of cancer diseases, skin diseases, and infectious diseases. Furthermore, since ultrasound can be used, it can reach deep areas, unlike light, and has the advantage that the treatment itself can be carried out easily, providing a new treatment system.

[0035] Furthermore, they are effective as antibacterial agents for SDT in the treatment of infectious diseases, and in particular, since the present electron-withdrawing group-bearing phenylchlorin derivatives or ethylene glycol derivatives or their metal complexes can ensure water solubility, they can be used not only as non-aqueous topical agents but also as aqueous topical preparations. Furthermore, while the anticancer drugs, antibacterial agents, antibiotics, and other drugs currently used in the treatment of cancer, skin diseases, and infectious diseases are prone to develop drug resistance, the SDT treatment system of the present invention, which uses ultrasound as the external energy, has a very unique advantage in that such drug resistance does not occur.

[0036] In addition, this compound group also shows an enhancing effect when NaCl, menthol, NaHCO3, EDTA, etc. are added, as in the case of the substance already proposed by the present inventor (Patent Document 3). [Brief description of the drawings]

[0037] [Figure 1] 1 is a graph showing the results of a phototoxicity test using cultured B16-F10 cells in Test Example 1. [Diagram 2] 1 is a graph showing the results of a phototoxicity test using cultured HDF cells in Test Example 1. [Diagram 3]1 is a graph showing the results of a phototoxicity test using cultured B16-F10 cells in Test Example 1. [Figure 4] 1 is a graph showing the results of a cytotoxicity test using B16-F10 in Test Example 2. [Diagram 5] 1 is a graph showing the results of a cytotoxicity test using HDF in Test Example 2. [Figure 6] 1 is a graph showing the results of a cytotoxicity test using B16-F10 in Test Example 2. [Figure 7] 1 is a graph showing the results of a cytotoxicity test using HDF in Test Example 2. [Figure 8] 1 is a graph showing the results of a cytotoxicity test using B16-F10 in Test Example 2. [Figure 9] 1 is a graph showing the results of a cytotoxicity test using B16-F10 in Test Example 2.

[0038] [Figure 10] 1 is a graph showing the results of an SDT activity test using B16-F10 in Test Example 3. [Figure 11] 1 is a graph showing the results of an SDT activity test using HDF in Test Example 3. [Figure 12] 1 is a graph showing the results of an SDT activity test using B16-F10 in Test Example 3. [Figure 13] 1 is a graph showing the results of an ultrasound irradiation test after administration of a compound to cancer-bearing animals in Test Example 4. [Figure 14] 1 is a graph showing the results of an ultrasound irradiation test after administration of a compound to cancer-bearing animals in Test Example 4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0039] The electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative represented by formula (I) or its metal complex or pharmacologically acceptable salt provided by the present invention is specifically an electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative of formula (I)-A type or formula (I)-B type (hereinafter, these may be collectively referred to simply as "chlorin derivatives"). This chlorin derivative, like the chlorin derivatives TONS503 and 504-Mn complex (Patent Document 3) that the present inventors have previously provided, has high affinity for cancer and high skin permeability, and is non-phototoxic. In addition, since it is water-soluble or fat-soluble, it can be uniformly dissolved or dispersed in, for example, injections or various ointment bases, and the stability of the ointment formulation itself is also extremely good. Furthermore, it can be uniformly dissolved or dispersed in formulations as water-soluble lotions, and the stability of the formulation itself is also extremely good.

[0040] Among such chlorin derivatives represented by formula (I), particularly, types A and B represented by the following formula (I):

[0041] [ka]

[0042] or a metal complex thereof, or a pharmacologically acceptable salt thereof, and specifically, the compounds represented by the following abbreviations.

[0043] Compound A : A phenylchlorin amino acid derivative having an electron-withdrawing group, in which in the formula (I) type B, X is [Chemical formula 2], Y is Cl, and R is an iminodiacetic acid residue.

[0044] Compound B : An electron-withdrawing group-bearing phenylchlorine ethylene glycol derivative in which, in the formula (I) type B, X is [Chemical formula 2], Y is Cl, and R is -NH(CH2CH2O)2OH.

[0045] Compound C: An electron-withdrawing group-bearing phenylchlorine ethylene glycol Mn metal complex, in which in the formula (I) type B, X is [Chemical formula 2], Y is Cl, R is -NH(CH2CH2O)2OH, and M is Mn.

[0046] Compound D : A phenylchlorin amino acid derivative having an electron-withdrawing group, in which in the formula (I) type B, X is [Chemical formula 2], Y is F, and R is an iminodiacetic acid residue.

[0047] Compound E : An electron-withdrawing group-bearing phenylchlorine ethylene glycol derivative in which, in the formula (I) type B, X is [Chemical formula 2], Y is F, and R is -NH(CH2CH2O)2OH.

[0048] Compound F : A chlorine ethylene glycol derivative of formula (I) type B, in which X is [Chemical formula 3] and R is -NH(CH2CH2O)2OH.

[0049] Compound G : Chlorine ethylene glycol Mn metal complex in which, in the formula (I) type B, X is [Chemical formula 3], R is -NH(CH2CH2O)2OH, and M is Mn.

[0050] In these phenylchlorin derivatives, R for Compound A or D may be not only an iminodiacetic acid residue but also an aspartic acid residue or a glutamic acid residue, and may be any amino acid having a polycarboxylic acid.

[0051] Furthermore, R for Compounds B, C, E, F and G may be not only two linked ethylene glycol residues but also one or three linked ethylene glycol residues.

[0052] On the other hand, Y may be not only Cl but also other halogen compounds exhibiting electron-withdrawing groups, namely Br, I, and F atoms, and may also be NH2, OH, and OCH3, which similarly exhibit electron-withdrawing groups. In short, all that is needed is a functional group that carries an electron-withdrawing group and eliminates phototoxicity, and among these, the ortho or para orientation of the phenyl group is the best. However, there is no need to be so strict about this.

[0053] Regarding the metal complexation, these synthesis procedures may be changed and the metal complexation may be carried out at the initial stage or at any stage during the synthesis. In short, as long as the target compound is finally obtained, there is no problem in forming a metal complex at any stage of the synthesis procedure. The contents of said patent are therefore incorporated herein by reference.

[0054] As a method for complexing, the compound obtained by the method described in the above patent is dissolved in a suitable organic solvent, and then the compound is reacted with an acetate or chloride of manganese or copper to prepare a metal complex of the phenylchlorin derivative or ethylene glycol derivative.

[0055] The metal salt and organic solvent used in the reaction are not particularly limited, and can be selected as long as they do not directly affect the reaction. Specifically, the metal salt can be an acetate or a metal chloride, of which metal chloride is preferably used, and the organic solvent can be an acetic acid, DMF, or other solvent, of which acetic acid is preferably used.

[0056] The reaction temperature and reaction time are not particularly limited, and the reaction may be carried out under heating at 40 to 80° C., preferably at about 55° C., for about 4 to 10 hours with stirring.

[0057] On the other hand, when the chlorin derivative is converted into an electron-withdrawing group-bearing phenyl derivative or an ethylene glycol derivative, the derivatization may be carried out not only at the initial stage of the synthetic procedure but also at any intermediate stage in the process. In short, as long as the target compound is finally obtained, there is no problem whether the compound is derivatized with an electron-withdrawing group-bearing phenyl derivatization or ethylene glycol derivatization at any stage of the synthetic procedure.

[0058] Some specific preparation methods are shown below.

[0059] Production Example 1: Preparation of dichlorophenylchlorin amino acid derivatives in which X is [Chemical formula 2], Y is chlorine, and R is an iminodiacetic acid residue in the B type of formula (I) (Preparation of Compound A)

[0060] (a) Preparation of CP-PH Photoprotoporphyrin IX dimethyl ester (P-MeB type) (10 g) and 2,4-dichlorophenylhydrazine HCl salt (5 g) were dissolved in pyridine (100 mL) and reacted with stirring for 1.5 hours. After the reaction, the reaction solution was added to chilled 16% acetic acid water, and the precipitate was filtered. The collected matter (CP-P-Me) was washed with water and air-dried. The air-dried product was dissolved in DMF (150 mL), 1.5M-NaOH (40 mL) was added, and the mixture was stirred at room temperature for 1.5 hours to hydrolyze. After hydrolysis, the hydrolyzate was precipitated by adding cold water, and the precipitate was filtered and the collected matter was washed several times with water to obtain about 12 g of hydrolyzate (CP-P-Na). The hydrolyzate obtained was suspended in 10% aqueous citric acid solution (800 mL) and stirred for 2.5 hours. After stirring, the mixture was collected by filtration, washed with water, and then dried to obtain about 10 g of CP-PH.

[0061] NMR and IR analysis of CP-PH 1 H-NMR (400 MHz, CDCl3) δ 9.72 (s, 1H), 9.54 (s, 1H), 8.61 (s, 1H), 8.43 (s, 1H), 7.74-7.99 (m, 3H), 7.03 (s, 1H), 6.03-6.13 (m, 2H), 4.13-4.38 (m, 4H), 3.69 (s, 6H), 3.46 (s, 3H), 3.18-3.36 (m, 4H), 1.26 (s, 3H) Gamma porphyrin = 1732, 2922, 2950 cm -1 , γ NH=N = 1510, 1593 cm -1

[0062] (b) Preparation of iminodiacetic acid dimethyl ester Methanol (50 mL) was added to iminodiacetic acid (10 g) and the mixture was stirred in a suspended state on an ice bath. Then, thionyl chloride (16 mL) was slowly added dropwise. After all the contents were added, the mixture was reacted at room temperature for 12 hours. After the reaction, ether was added and the precipitate was filtered, and the collected matter was washed thoroughly with ether. The washed solid was collected and dried to obtain about 11 g of iminodiacetic acid dimethyl ester hydrochloride (IDA(OMe)2 HCl salt).

[0063] NMR analysis of IDA(OMe)2·HCl salt 1 H-NMR (500 MHz, DMSO-D6) δ 10.05 (s, 1H), 4.01 (s, 4H), 3.74 (s, 6H)

[0064] (c) Preparation of the target compound (Compound A) CP-PH (5.0 g) (781 g / mol) obtained in (a) above and EDC / HCl (10 g) (155.25 g / mol) were dissolved in N,N-dimethylacetamide (100 mL). Then, IDA(OMe)2·HCl salt (6.0 g) obtained in (b) was added and reacted for 2.5 hours. Then, an additional 4.0 g of EDC / HCl was added and reacted for 1.5 hours. After the reaction, the mixture was poured into cold water to obtain a precipitate. The precipitate was collected by filtration, washed with water and EtOH, and dried to obtain approximately 5 g of CP-P-IDA(OMe)2. The obtained CP-P-IDA(OMe)2 was suspended in EtOH (300 mL), 1.5-M-NaOH (50 mL) was added, and hydrolysis was performed at room temperature for 1.5 hours. To the obtained hydrolyzate, EtOH / AcOEt mixed solution (1:1) (500 mL) was added to obtain a precipitate. The precipitate was collected by filtration, washed with EtOH / AcOEt mixed solution and EtOH, and then dried to obtain 4 g of the target compound A.

[0065] NMR and IR analysis of Compound A (Mw: 1,071.78) 1H-NMR (400 MHz, CDCl3) δ 9.73 (s, 1H), 9.60 (s, 1H), 8.70 (s, 1H), 8.42 (s, 1H), 8.11 (br, 1H), 7.75 (d, J = 8.4 Hz, 2H), 7.37-7.43 (m, 2H), 7.18 (br, 1H), 6.16 (d, J = 18.4 Hz, 1H), 6.05 (d, J = 12.0 Hz, 1H), 4.28-4.39 (m, 2H), 4.18-4.24 (m, 2H), 3.66-3.71 (m, 8H), 3.54 (s, 3H), 3.46 (s, 3H), 3.35 (s, 3H), 3.27 (t, J = 6.8 Hz, 2H), 3.21 (t, J = 4.4 Hz, 2H), 1.44 (s, 3H) Gamma porphyrin = 2956 cm -1 , γ COO = 1732 cm -1 , γ CONRR’ = 1594 cm -1 ,γ NH=N = 1510 cm -1 This structure was also supported by UV spectroscopy.

[0066] Preparation Example 2: In the B type of formula (I), X is [Chemical formula 2], Y is chlorine, and R is -NHCH 2 CH 2 OCH 2 CH 2 Preparation of dichlorophenylchlorin ethylene glycol derivatives (preparation of Compound B) CP-PH (3.0 g) obtained in (a) of the previous Production Example 1 was used, and EDC / HCl (6.0 g) was added to it and dissolved in N,N-dimethylacetamide. Then, 2-(2-aminoethoxy)ethanol (DEGA) (2.6 mL) was added and reacted for 1 hour. EDC / HCl (1.0 g) was further added and reacted for 3 hours. After the reaction, the reaction product was added to cold water to obtain a precipitate. The resulting reaction product was washed with water and then EtOH, and dried in a desiccator to obtain 2.8 g of the target Compound B.

[0067] NMR and IR analysis of Compound B (Mw: 927.93) 1 H-NMR (400 MHz, CDCl3) δ 9.73 (s, 1H), 9.53 (s, 1H), 8.52 (s, 1H), 8.42 (s, 1H), 7.93 (br, 1H), 7.60-7.77 (m, 2H), 7.39-7.44 (m, 2H), 6.98 (br, 1H), 6.11 (d, J = 17.6 Hz, 1H), 6.03 (d, J = 11.6 Hz, 1H), 4.22-4.34 (m, 4H), 4.12 (q, J = 6.9 Hz, 4H), 3.20-3.72 (m, 25H), 1.24 (s, 3H) Gamma OH = 3341 cm -1 , γ porphyrin = 1736, 2922, 2955 cm -1 , γ CONR = 1593 cm -1 , γ NH=N = 1510 cm -1 ,γ R-O-R’ = 1115, 1239 cm -1 This structure was also supported by UV spectroscopy.

[0068] Preparation Example 3: In the B type of formula (I), X is [Chemical formula 2], Y is chlorine, and R is -NHCH 2 CH 2 OCH 2 CH 2 Preparation of dichlorophenylchlorin ethylene glycol Mn complex where OH, M is Mn (preparation of Compound C) Compound B (1.0 g) obtained in the previous Production Example 2 was used, and after dissolving it in 30 mL of DMF, Mn(OAc)2 was added and the mixture was heated and boiled for 5 hours to perform manganization. After the reaction, the reaction product was added to cold water to obtain a precipitate. The reaction product was washed with water and then ethanol, and dried in a desiccator to obtain 0.8 g of the target compound C. The stability of Compound C was confirmed by HPLC, and it was found to be stable. Inductively Coupled Plasma (ICP) analysis also confirmed that Mn ions were 100% coordinated.

[0069] Production Example 4: Preparation of difluorophenyl chlorin amino acid derivatives in which X is [Chemical formula 2], Y is fluorine, and R is an iminodiacetic acid residue in the B type of formula (I) (preparation of Compound D) (a) Preparation of FP-PH Photoprotoporphyrin IX dimethyl ester (P-MeB type) (10 g) and 2,4-difluorophenylhydrazine HCl salt (5 g) were dissolved in pyridine (100 mL) and reacted with stirring for 1.5 hours. After the reaction, the reaction solution was added to cooled 16% acetic acid water and the precipitate was filtered. The collected matter (FP-P-Me) was washed with water and air-dried to obtain FP-PH. The following treatments were performed in the same manner as in Production Example 1 to obtain the target compound D.

[0070] Production Example 5: In the B type of formula (I), X is [Chemical formula 2], Y is fluorine, and R is -NHCH 2 CH 2 OCH 2 CH 2 Preparation of dichlorophenylchlorin ethylene glycol derivatives (preparation of Compound E) The target compound, Compound E, was obtained by treating FP-PH (3.0 g) obtained in (a) of Production Example 4 in the same manner as in Production Example 2.

[0071] Preparation Example 6: In the B type of formula (I), X is [Chemical formula 3] and R is -NHCH 2 CH 2 OCH 2 CH 2 Preparation of OH chlorine ethylene glycol derivatives (preparation of Compound F) (a) Preparation of PG-P-Me PG-P-Me was obtained by carrying out the reaction, post-treatment, and other operations in the same manner as described in the manufacturing example of the previous Japanese Patent Application No. 2010-238359 (Patent No. 5651426). (b) Preparation of PG-P-DEGA According to the method of the above Production Example 2, PG-P-Me was hydrolyzed to give PG-PH, and the rest of the procedure was carried out in the same manner as in Production Example 2 to obtain Compound F (PG-P-DEGA).

[0072] Preparation Example 7: In the B type of formula (I), X is [Chemical formula 3] and R is -NHCH 2 CH 2 OCH 2 CH 2 Preparation of chlorine ethylene glycol Mn complex (OH, M=Mn) (Preparation of Compound G) Compound F was obtained in the same manner as in Preparation Example 6, and then the compound was converted into a Mn complex to obtain the desired Compound G (PG-P-DEGA Mn complex).

[0073] In the present invention, these phenylchlorin derivatives or ethylene glycol derivatives or metal complexes thereof are formulated as injectable preparations for SDT such as cancer disease therapeutic agents, skin disease therapeutic agents, or external preparations such as antibacterial agents. Injectable preparations for treating cancer diseases are prepared using osmotic pressure and pH adjusters, and sublingual preparations are prepared using appropriate gelling agents, etc., and topical preparations for treating skin diseases or antibacterial agents are prescribed in the form of non-aqueous ointments, aqueous ointments, lotions, etc.

[0074] In the present invention, the amount of the phenylchlorin derivative or ethylene glycol derivative or its metal complex represented by formula (I) contained in these preparations may be an amount sufficient for the active ingredient phenylchlorin derivative or ethylene glycol derivative or its metal complex to reach and remain at the affected site, be absorbed percutaneously, accumulate at the diseased site, and kill target cells or bacteria by ultrasonic irradiation. According to the study by the present inventors, it was found that a sufficient effect can be obtained if the amount is 0.1 to 20% by weight based on the weight of the preparation.

[0075] If the amount was less than 0.1% by weight, the desired therapeutic effect could not be achieved, and even if the amount was more than 20% by weight, no further effect could be obtained. The amount of the active ingredient to be added depends on the type of active ingredient contained, and cannot be generally determined. Furthermore, since the stability of the active ingredient contained is not significantly affected by the concentration of the active ingredient or the base used, the amount can be varied in various ways within the above content range according to the application.

[0076] When the preparation of the present invention obtained as described above is used for SDT, the preparation can be administered by drip injection, sublingual administration, suppository administration or local injection into the affected area for various cancers, or by direct application to the affected area for actinic keratosis, inflammatory keratosis, epidermal cancer, infectious diseases and the like, whereby the phenylchlorin derivative, ethylene glycol derivative or metal complex thereof can be effectively accumulated, and then the affected area can be irradiated with ultrasound or the like to effectively treat the disease.

[0077] In application of topical preparations such as ointments and lotions, the phenylchlorin derivatives or ethylene glycol derivatives or their metal complexes provided by the present invention have good permeability to affected areas of the skin without the need for tape stripping, and therefore have the advantage that in SDT for treating skin diseases, the treatment itself can be carried out easily without imposing a burden on the patient. When applying topical agents such as ointments and lotions, it is more effective to keep the application site sealed in order to obtain the ODT effect (Occlusive Dressing Technique).

[0078] Furthermore, the phenylchlorin derivatives or ethylene glycol derivatives or metal complexes thereof of the present invention have been found to be effective as antibacterial agents for SDT against various infectious diseases, including infectious diseases in not only humans but also various animals, and can be used for the treatment of SDT against fungal infections such as Malassezia otitis externa, nail infections, and athlete's foot in dogs and cats caused by yeast-like fungi, various infectious diseases caused by bacteria such as MRSA, Pseudomonas aeruginosa, and Escherichia coli, dental infections such as pyorrhea caused by periodontal disease bacteria, and other infectious diseases and warts caused by tuberculosis bacteria or viruses.

[0079] Various types of ultrasound can be used for irradiating the affected area with ultrasound after application of the preparation containing the phenylchlorin derivative or ethylene glycol derivative or its metal complex provided by the present invention. Among them, it is more effective to use a medical ultrasound irradiator or the like.

[0080] Thus, the injectable formulation, ointment formulation, lotion formulation, etc. of the present invention can be injected or applied to the diseased area, allowing the active ingredient to accumulate in the affected area or be absorbed percutaneously, and then the diseased area can be irradiated with ultrasound, thereby effectively treating the cancer disease, skin disease, or infectious disease. EXAMPLES

[0081] The present invention will be described in detail below with reference to specific formulation examples, test examples, etc., but the present invention is not limited to these.

[0082] Test Example 1: Phototoxicity test of phenylchlorin derivatives (Compound A, Compound B, Compound F, and Compound G) The phenylchlorin derivatives or ethylene glycol derivatives or their metal complexes of the present invention are desired to be photoinsensitive, so the phototoxicity of the derivatives against light treatment was examined by the phototoxicity against treatment with LEDs of 660 nm and 405 nm. At the same time, the aforementioned substance TONS504 (Patent Document 3) was used as a light positive control group.

[0083] <Experimental Procedure> (1) 1 x 10 in a 96-well plate 3 Mouse melanoma cells (B10-F10) or human fibroblast cells (HDF) were seeded at 10 cells / well and cultured in a 5% carbon dioxide incubator. (2) Each compound was dissolved in DMSO to a concentration of 100 μM and diluted with medium to a final concentration of 1 μM. (3) The medium containing the compound was added during the medium change, and after 5 hours, the cells were washed twice with 1x PBS (phosphate-buffered saline), after which PBS was added and LED light was irradiated under the following conditions.

[0084] <Irradiation conditions> Wavelength: 405nm or 660nm ·Irradiance: 46.1mW / cm 2 Irradiation time: 6 minutes

[0085] (4) 48 hours after irradiation, the cells were washed twice with 1×PBS, and then 50 μL of 0.5% crystal violet solution was added. (5) After incubation at room temperature for 15 minutes, the plate was washed four times with 100 μL / well of tap water. (6) After drying at room temperature for 1 hour, 100 μL / well of 1% SDS was added. After confirming that the crystal violet had dissolved, the absorbance (abs: 570 nm) was measured using a microplate reader to calculate the survival rate.

[0086] <Result> The results are shown in Figures 1, 2 and 3. As is clear from the results shown in the figures, the phenylchlorin derivatives or ethylene glycol derivatives and their Mn complexes of the present invention were confirmed to be stable, i.e., to have no phototoxicity, in the light irradiation experiments using B10-F10 cells and HDF cells. On the other hand, photosensitization was observed in the control groups, which were the precursor substances TONS504 and Compound F.

[0087] Test Example 2: Cytotoxicity test of phenylchlorin derivatives or ethylene glycol derivatives and their Mn metal complexes (Compound A, Compound B, Compound F, and Compound G) It is desirable that the phenylchlorin derivatives or ethylene glycol derivatives and their Mn metal complexes of the present invention are non-cytotoxic in the dark. This was confirmed by the following test.

[0088] <Experimental Procedure> (1) 2.5 × 10 in a 96-well plate 3 B10-F10 cells and HDF cells were seeded separately at 10 cells / well and cultured in a 5% carbon dioxide incubator. (2) Compound A, Compound B, Compound F, and Compound G were measured out and their concentrations were adjusted with DMSO to 10 mM, 30 mM, 10 mM, 3 mM, 1 mM, 0.3 mM, 0.1 mM, 0.03 mM, and 0.01 mM, respectively. (3) DMSO was diluted with DEME medium to 1%, and the final concentrations were adjusted to 1000 μM, 300 μM, 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, and 0.1 μM, and added when the medium was replaced. (4) 24 hours after addition, the cells were washed twice with 1x PBS, and the medium was added and incubated. (5) After 24 hours, the plates were washed twice with 1x PBS and 50 μL of 0.5% crystal violet solution was added. (6) After incubation at room temperature for 15 minutes, the plate was washed four times with 100 μL / well of tap water. (7) After drying at room temperature for 1 hour, 100 μL / well of 1% SDS was added. After confirming that the crystal violet had been dissolved, the absorbance (abs: 570 nm) was measured using a microplate reader to calculate the survival rate.

[0089] <Result> The results are shown in Figures 4, 5, 6, 7, 8 and 9. As is clear from the results shown in the figures, the phenylchlorin derivatives or ethylene glycol derivatives of the present invention and their Mn metal complexes (Compound A, Compound B and Compound G) had no cytotoxicity at 10 μM or less. Regarding Compound A, a small amount of precipitation appeared to occur at concentrations of 100 μM or higher.

[0090] Test Example 3: Ultrasonic activity test of phenylchlorin derivatives and ethylene glycol derivatives (Compound A, Compound B, Compound F, and Compound G) The phenylchlorin derivatives or ethylene glycol derivatives and their Mn metal complexes of the present invention are preferably sensitized to ultrasound. Therefore, the sensitivity to ultrasound treatment was examined as follows.

[0091] <Experimental Procedure> (1) 1×10 in a Φ6 petri dish 5 B10-F10 cells and HDF cells were seeded separately at 10 cells / well and cultured in a 5% carbon dioxide incubator. (2) Each compound was dissolved in DMSO to a concentration of 100 μM and diluted with medium to a final concentration of 1 μM. (3) The medium containing the compound was added during the medium change, and after 5 hours, ultrasonic irradiation was performed under the following conditions.

[0092] <Irradiation conditions> Probe L (effective irradiation area: 5cm 2 ) Irradiation time: 4 minutes Duty cycle: 20% Frequency: 1MHz ·Irradiation amount: 0.4W / cm 2

[0093] (4) 24 hours after irradiation, the cells were washed twice with 1x PBS, 500 μL of 1x Trypsin was added, and the cells were incubated for 3 minutes. Then, the cells were physically detached and 1 mL of medium was added. (5) 30 μL of the cell suspension was combined with 30 μL of 0.4% Trypan blue, and the number of live cells was counted using a hemocytometer to calculate the survival rate.

[0094] <Result> The results are shown in Figures 10 and 12. As is clear from the results shown in the figures, the phenylchlorine chlororin derivative or ethylene glycol derivative and its metal complex of the present invention were confirmed to have a cell destruction effect in an ultrasound irradiation experiment using B16-F10 cells and HDF cells. In the figure, US indicates the result of ultrasonic irradiation, A indicates Compound A, A+US indicates the result of ultrasonic irradiation on Compound A, B indicates the result of ultrasonic irradiation on Compound B, B+US indicates the result of ultrasonic irradiation on Compound B, F+US indicates the result of ultrasonic irradiation on Compound F, and G+US indicates the result of ultrasonic irradiation on Compound G.

[0095] The actual therapeutic effect of SDT on the preparations containing the novel phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes provided by the present invention will be described below.

[0096] Test Example 4: Ultrasound irradiation test using tumor-bearing animals <Method> Six-week-old female mice of the Balb / c strain were used. Tumor-bearing mice were inoculated with 1 × 10 EMT-6 tumor cells. 7 The mice were prepared by inoculating each mouse into the dorsal region of the back. Six days after inoculation, the phenylchlorin derivative or ethylene glycol derivative of the present invention and its metal complex (Compound B and Compound G) were administered at 1 mg / kg, 5 mg / kg, or 10 mg / kg, respectively, via the tail vein, and ultrasonic irradiation was performed for 5 minutes 4 hours after administration.

[0097] <Ultrasonic irradiation conditions> -Ito Ultrasonics Co., Ltd. equipment: UST770 Frequency: 1MHz ·Irradiation amount: 2W / cm 2 Duty cycle: 50%

[0098] <Result> The results are shown in Figures 13 and 14. As is clear from the results shown in the figures, it was confirmed that the phenylchlorin derivative or ethylene glycol derivative and its metal complex of the present invention have a tumor suppressing effect, and in particular, in the case of ultrasonic irradiation using Compound B and Compound G, a remarkable effect was observed when ultrasonic irradiation was performed for 5 minutes 4 hours after administration of 1 mg, 5 mg, and 10 mg of the drug. [Industrial Applicability]

[0099] As described above, the present invention provides phenylchlorin derivatives or ethylene glycol derivatives carrying electron-withdrawing groups and their metal complexes for use in ultrasonic chemotherapy (SDT). The phenylchlorin derivatives or ethylene glycol derivatives carrying electron-withdrawing groups and their metal complexes provided by the present invention have extremely good delivery to the affected area and skin permeability (transdermal absorbability) when made into injections or preparations for use in cancer, sublingual preparations, or ointments for use on the skin. Therefore, the phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes carrying an electron-withdrawing group are selectively accumulated at the site of cancer disease, and by applying the same to the site of a skin disease such as actinic keratosis, inflammatory keratosis, epidermal cancer, or an infectious disease, e.g., papilloma, the phenylchlorin derivatives or ethylene glycol derivatives and their metal complexes carrying an electron-withdrawing group are effectively accumulated at the site of the disease, and cancer or skin disease can be effectively treated by ultrasonic irradiation.

[0100] Furthermore, as an antibacterial agent for SDT, it exhibits antibacterial activity against various bacteria. While antibiotics currently used to treat infectious diseases are prone to develop drug resistance, the SDT diagnostic and therapeutic system of the present invention has the advantage that such drug resistance does not occur, making it extremely specific and of great medical value. Furthermore, past test results have shown that when the present electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative is complexed with Mn, its water solubility increases and it is also effective as an MRI contrast agent, and has the advantage that after administration of the present electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative and its Mn metal complex, the affected area can be identified by MRI and then SDT irradiation treatment can be performed.

Claims

1. The following formula (I): 【Chemistry 1】 [In the formula, X is of the following formula: 【Chemistry 2】 Or the following formula: 【Chemistry 3】 (In the above formula, Y represents an electron-withdrawing group, a halogen atom, NH ２ , OH, or OCH ３ is a group represented by: R is -N(CH ２ COONa) ２ [which represents the residue of iminodiacetic acid], an aspartic acid residue or a glutamic acid residue, or -NH(CH ２ CH ２ O) ｎ H (n represents an integer of 1 to 3), M represents Mn, Fe or Cu; Z represents a ligand and is OAc or Cl. or a pharmacologically acceptable salt thereof.

2. Formula (I) is the following formula (I)-A type: 【Chemistry 4】 (In the formula, X, R, Y, M, Z and n are the same as defined above.) 2. The metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 1, which is represented by the following formula: or a pharmacologically acceptable salt thereof.

3. Formula (I) is of the following formula (I)-B type: 【Chemistry 5】 (In the formula, X, R, Y, M, Z and n are the same as defined above.) 2. The metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 1, which is represented by the following formula: or a pharmacologically acceptable salt thereof.

4. The following formula (II): 【Chemistry 6】 【Chemistry 7】 [In the formula, X is of the following formula: 【Chemistry 8】 Or the following formula: 【Chemistry 9】 (In the above formula, Y represents an electron-withdrawing group, a halogen atom, NH ２ , OH, or OCH ３ is a group represented by: R is -N(CH ２ COONa) ２ [which represents the residue of iminodiacetic acid], an aspartic acid residue or a glutamic acid residue, or -NH(CH ２ CH ２ O) ｎ H (n represents an integer of 1 to 3). or a pharmacologically acceptable salt thereof.

5. A metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to any one of claims 1 to 3, or a pharmacologically acceptable salt thereof; The electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 4, or a pharmacologically acceptable salt thereof. A cancer disease treatment agent for sonodynamic therapy (SDT), comprising as an active ingredient

6. A metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to any one of claims 1 to 3, or a pharmacologically acceptable salt thereof; The electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 4, or a pharmacologically acceptable salt thereof. A skin disease treatment agent for sonodynamic therapy (SDT), comprising as an active ingredient

7. A metal complex of the electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to any one of claims 1 to 3, or a pharmacologically acceptable salt thereof; The electron-withdrawing group-bearing phenylchlorin derivative or ethylene glycol derivative according to claim 4, or a pharmacologically acceptable salt thereof. An antibacterial agent for sonodynamic therapy (SDT), comprising as an active ingredient

8. The cancer disease therapeutic agent for sonodynamic therapy (SDT) according to claim 5, which is in the form of an injection.

9. The cancer disease therapeutic agent, skin disease therapeutic agent or antibacterial agent for sonodynamic therapy (SDT) according to claim 5, 6 or 7, which is in the form of an ointment.

10. The cancer disease therapeutic agent, skin disease therapeutic agent, or antibacterial agent for sonodynamic therapy (SDT) according to claim 5, 6, or 7, which is in the form of a lotion.

11. The cancer disease therapeutic agent, skin disease therapeutic agent, or antibacterial agent for sonodynamic therapy (SDT) according to claim 5, 6, or 7, which is in the form of a sublingual tablet.

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