Use of diketone compounds in photodynamic therapy or photodynamic diagnosis

Abstract

To provide diketone compounds for use in photodynamic therapy or photodynamic diagnosis of diseases or for use in skin cosmetic treatment by photodynamic therapy.SOLUTION: Provided are a compound of formula (I), a pharmaceutically acceptable salt or ester of the compound, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form of the compound, a metabolite of each thereof, or any combination or mixture thereof.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure is in the pharmaceutical field and relates to the use of diketone compounds in photodynamic therapy or photodynamic diagnosis. [Background technology]

[0002] Photodynamic therapy is a photochemotherapy that induces oxygen based on sensitization by photosensitizers to generate reactive oxygen species (ROS) represented by singlet oxygen, and kills tumor cells or pathogenic microorganisms, etc., to achieve the purpose of treating diseases. Photodynamic therapy is a novel treatment method used in several clinical treatments, including the treatment of tumors, genital warts, acne, and port-wine stains. Compared with traditional treatments for tumors, photodynamic therapy has effective advantages such as convenience of surgery, high selectivity, and rare side effects. In addition, photodynamic therapy can also be used in skin cosmetics. Photodynamic diagnosis refers to the characteristic spectrum of photosensitizers that have affinity for diseased tissues, and is used to diagnose diseases under the action of light. Photodynamic diagnosis has been used in the diagnosis of tumors, genital warts, acne, and port-wine stains. [Prior art documents] [Non-patent literature]

[0003] [Non-Patent Document 1] Remington's Pharmaceutical Sciences(1990) Summary of the Invention [Problem to be solved by the invention]

[0004] The basic elements of photodynamic therapy or photodynamic diagnosis include oxygen, photosensitizer, and light. Photosensitizer is the key to photodynamic therapy or photodynamic diagnosis. An ideal photosensitizer is non-toxic, inexpensive, easily available, has good photochemical properties, and can be rapidly metabolized. At present, the photosensitizer based on the structure of porphyrin and / or phthalocyanine is the most widely used in clinical research. However, due to the large cyclic π-conjugated system of this type of photosensitizer, the π-π mutual effect between the molecules of the photosensitizer is strong, which easily causes the porphyrin and / or phthalocyanine molecules to aggregate and reduce their photochemical efficiency, greatly limiting the clinical application of the photosensitizer. Therefore, exploring new photosensitizer molecules with good photochemical properties will help promote the clinical application of photodynamic therapy or photodynamic diagnosis, and will not only have important research significance, but also have applicability value. [Means for solving the problem]

[0005] Through thorough research, the inventors of the present disclosure have inventively discovered that certain diketone compounds exhibit effective antitumor, antimicrobial and immune-enhancing effects by inducing oxygen to generate singlet oxygen and / or peroxy free radicals and other reactive oxygen species after being excited by light.Therefore, they can be used in the photodynamic treatment or photodynamic diagnosis of diseases, and can also be used in skin beauty by photodynamic treatment, and therefore the following disclosure is provided.

[0006] According to one aspect, the present invention provides the use of a compound of formula (I), or a pharma- ceutically acceptable salt or ester, prodrug, stereoisomer, hydrate, solvate or crystalline form of said compound, or a metabolite of each thereof, or any combination or mixture thereof, in the preparation of a medicament or reagent for use in the photodynamic treatment or photodynamic diagnosis of a disease or for use in skin cosmetology by photodynamic therapy.

[0007] [ka]

[0008] In formula (I), R 1 and R 2 are each independently a linear or branched alkyl group having 1 to 6 carbon atoms, a halogenated linear or branched alkyl group having 1 to 6 carbon atoms, a hydroxyl group, or an aryl group having 6 to 14 carbon atoms, a 5- to 6-membered heteroaryl group, a 5- to 6-membered heterocyclyl group, and a 3- to 6-membered cycloalkyl group, each of which is optionally substituted with one or more (e.g., 2, 3, 4, or 5) identical or different substituents, each of which is independently a hydroxyl group, a carboxyl group, a sulfonic group, a halogen (e.g., fluorine, chlorine, bromine, or iodine) atom, an amino group, a mercapto group, a nitro group, -C(O)-(linear or branched alkyl group having 1 to 4 carbon atoms)-S(O)2O-linear or branched alkyl group, or -O-(linear or branched alkyl group having 1 to 4 carbon atoms).

[0009] In some embodiments, the straight or branched chain alkyl having 1 to 6 carbon atoms is a straight or branched chain alkyl having 1 to 4 carbon atoms.

[0010] In some embodiments, the straight or branched alkyl having 1 to 4 carbon atoms is methyl, ethyl, propyl (n-propyl), isopropyl, butyl (n-butyl), s-butyl (sec-butyl), isobutyl, or t-butyl (tert-butyl).

[0011] In some embodiments, the aryl having 6 to 14 carbon atoms is phenyl or naphthyl.

[0012] In some embodiments, the halogenated linear or branched alkyl having 1 to 6 carbon atoms is a fluorinated, chlorinated, brominated, or iodinated linear or branched alkyl having 1 to 6 carbon atoms. In some embodiments, the halogenated linear or branched alkyl having 1 to 6 carbon atoms is a halogenated linear or branched alkyl having 1 to 4 carbon atoms, such as fluoromethyl, bromomethyl, chloromethyl, fluoroethyl, bromoethyl, or chloroethyl.

[0013] As used herein, the term "halogenated" includes substitution with one or more (e.g., 2, 3, 4, or 5) halogen (e.g., fluorine, chlorine, bromine, or iodine) atoms.

[0014] In some embodiments, fluoromethyl is -CH2F, -CHF2, or -CF3. In some embodiments, bromomethyl is -CH2Br, -CHBr2, or -CBr3.

[0015] In some embodiments, chloromethyl is -CH2Cl, -CHCl2, or -CCl3.

[0016] In some embodiments, fluoroethyl is monofluoroethyl, difluoroethyl, trifluoroethyl, tetrafluoroethyl, or perfluoroethyl.

[0017] In some embodiments, bromoethyl is monobromoethyl, dibromoethyl, tribromoethyl, tetrabromoethyl, or perbromoethyl.

[0018] In some embodiments, chloroethyl is monochloroethyl, dichloroethyl, trichloroethyl, tetrachloroethyl, or perchloroethyl.

[0019] In some embodiments, the 5-6 membered heteroaryl contains 1-3 ring atoms selected from nitrogen, oxygen, and sulfur, In some embodiments, the 5-6 membered heteroaryl is furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, imidazolyl, pyridyl, pyrimidinyl, pyridazinyl, or pyrazinyl.

[0020] In some embodiments, the 5-6 membered heterocyclyl contains 1-3 ring atoms of nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocyclyl is aziridinyl, diaziridinyl, azetidinyl, dioxanyl, dioxolane, tetrahydrofuranyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, tetrahydrothienyl, piperidinyl, piperazinyl, morpholinyl, hexahydropyrimidinyl, or hexahydropyridazinyl. When the heteroaryl or heterocyclyl contains multiple heteroatoms, the multiple heteroatoms may be the same or different.

[0021] In some embodiments, R 1 is a straight or branched chain alkyl having 1 to 4 carbon atoms (eg, methyl or ethyl), or an aryl having 6 to 14 carbon atoms (eg, phenyl).

[0022] In some embodiments, R 2 is a straight or branched chain alkyl having 1 to 4 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl, butyl, or t-butyl), a halogenated straight or branched chain alkyl having 1 to 4 carbon atoms (e.g., bromomethyl or bromoethyl), or hydroxy.

[0023] In some embodiments, R 1 is a straight or branched alkyl having 1 to 4 carbon atoms (e.g., methyl or ethyl), and R 2is a straight or branched chain alkyl having 1 to 4 carbon atoms (eg, methyl, ethyl, propyl, isopropyl, butyl, or t-butyl).

[0024] In some embodiments, R 1 is a straight or branched alkyl having 1 to 4 carbon atoms (e.g., methyl or ethyl) or phenyl, and R 2 is a hydroxyl group.

[0025] In some embodiments, R 1 is a straight or branched alkyl having 1 to 4 carbon atoms (e.g., methyl or ethyl), and R 2 is a halogenated straight or branched chain alkyl having 1 to 4 carbon atoms (eg, bromomethyl).

[0026] In some embodiments, R 1 and R 2 Each of is phenyl, and the phenyl is optionally substituted with one or more substituents which can be the same or different.

[0027] In some embodiments, R 1 and R 2 each is independently a nitrogen-containing 5-6 membered heteroaryl, and the nitrogen-containing 5-6 membered heteroaryl is optionally substituted with one or more substituents, which can be the same or different.

[0028] For purposes of this specification, the phrase "optionally substituted with" means that it may or may not be substituted.

[0029] In some embodiments, R 1 and R 2 are the same functional group.

[0030] In some embodiments, R 1and R 2 is a separate functional group.

[0031] In some embodiments, the compound is selected from Tables 1 and 2 below.

[0032] [Table 1]

[0033] [Table 2]

[0034] In some embodiments, the compound, a pharma- ceutically acceptable salt or ester, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form of the compound, a metabolite of each thereof, or any combination or mixture thereof, is used as a photosensitizer for photodynamic therapy or photodynamic diagnosis.

[0035] In some embodiments, photodynamic therapy comprises the steps of: Step I of administering to a subject a compound according to any of the above, a pharma- ceutically acceptable salt or ester, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form of said compound, a metabolite of each thereof, or any combination or mixture thereof; Step II of irradiating the target with light; The method is carried out by a method comprising:

[0036] In some embodiments, in step I, the compound, a pharma- ceutically acceptable salt or ester of the compound, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form, a metabolite of each thereof, or any combination or mixture thereof, is applied to the subject by a suitable method (e.g., by drip, oral administration, perfusion, smear, or injection).

[0037] In some embodiments, the compound, a pharma- ceutically acceptable salt or ester of the compound, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystal form, a metabolite of each of them, or any combination or mixture thereof in step I is used in the form of a solution. In some embodiments, the solution is an aqueous solution. In some embodiments, the solution is a solution using saline as a solvent. In some embodiments, the concentration of the solution is 0.5 weight percent (wt%) to 3 weight percent (e.g., 0.5 weight percent to 1 weight percent, 1 weight percent to 2 weight percent, or 2 weight percent to 3 weight percent).

[0038] In step II, single wavelength light or mixed light can be used.

[0039] In some embodiments, at least a portion of the wavelengths of the light in step II are in the range of 10 nm to 1 mm, for example, one or more of the ranges of 10 nm to 380 nm, 380 nm to 780 nm, 780 nm to 3 μm, 3 μm to 30 μm, and 30 μm to 1 mm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 400 nm to 480 nm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 450 nm to 480 nm.

[0040] In some embodiments, the irradiation in step II is performed for 1 second to 12 hours, for example, 1 second to 10 seconds, 10 seconds to 100 seconds, 100 seconds to 200 seconds, 200 seconds to 500 seconds, or 500 seconds to 900 seconds, for example, 1 minute to 10 minutes, 10 minutes to 30 minutes, or 30 minutes to 60 minutes, for example, 1 hour to 2 hours, 2 hours to 5 hours, or 5 hours to 12 hours.

[0041] In some embodiments, the irradiation in step II is from 1 to 2000 mW / cm 2 , e.g., 1 to 15 mW / cm 2 , 15mW / cm 2 ~50mW / cm 2 , 50mW / cm 2~100mW / cm 2 , 100mW / cm 2 ~200mW / cm 2 , 200mW / cm 2 ~300mW / cm 2 , 300mW / cm 2 ~500mW / cm 2 , 500mW / cm 2 ~1000mW / cm 2 , 1000mW / cm 2 ~1500mW / cm 2 , or 1500mW / cm 2 ~2000mW / cm 2 The measurement is performed at a light intensity of .

[0042] In some embodiments, step II comprises irradiating the subject's body or surface with light at the desired treatment site (eg, the site where the lesion will occur) or irradiating the subject's entire body with light.

[0043] According to another aspect, the present disclosure provides a method for diagnosing or treating a disease in a subject comprising using a compound, a pharma- ceutically acceptable salt or ester, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form according to any of the above, a metabolite of each thereof, or any combination or mixture thereof, as a photosensitizer.

[0044] In some embodiments, the method comprises step I and step II as defined in any one of the above.

[0045] According to another aspect, the present disclosure provides a method for performing skin cosmetic treatment of a subject, comprising using a compound according to any of the above, a pharma- ceutically acceptable salt or ester, a prodrug, a stereoisomer, a hydrate, a solvate or a crystalline form of the compound, a metabolite of each of them, or any combination or mixture thereof, as a photosensitizer. In some embodiments, the skin cosmetic treatment is for non-therapeutic purposes.

[0046] In some embodiments, the method comprises the steps of: Step I' of applying to the skin of a subject a compound according to any of the above, a pharma- ceutically acceptable salt or ester of said compound, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form of said compound, a metabolite of each thereof, or any combination or mixture thereof; A process II' of irradiating the skin with light; In step II', single wavelength light or mixed light can be used.

[0047] In some embodiments, at least a portion of the wavelengths of the light in step II' are in the range of 10 nm to 1 mm, for example, one or more of the ranges of 10 nm to 380 nm, 380 nm to 780 nm, 780 nm to 3 μm, 3 μm to 30 μm, and 30 μm to 1 mm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 400 nm to 480 nm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 450 nm to 480 nm.

[0048] In some embodiments, the irradiation in step II' is performed for 1 second to 12 hours, for example, 1 second to 10 seconds, 10 seconds to 100 seconds, 100 seconds to 200 seconds, 200 seconds to 500 seconds, or 500 seconds to 900 seconds, for example, 1 minute to 10 minutes, 10 minutes to 30 minutes, or 30 minutes to 60 minutes, for example, 1 hour to 2 hours, 2 hours to 5 hours, or 5 hours to 12 hours.

[0049] In some embodiments, in step II', the irradiation is at 1 to 2000 mW / cm 2 , e.g., 1 to 15 mW / cm 2 , 15mW / cm 2 ~50mW / cm 2 , 50mW / cm 2 ~100mW / cm 2 , 100mW / cm 2 ~200mW / cm 2 , 200mW / cm 2 ~300mW / cm 2 , 300mW / cm 2 ~500mW / cm 2 , 500mW / cm2 ~1000mW / cm 2 , 1000m 2 ~1500mW / cm 2 , or 1500mW / cm 2 ~2000mW / cm 2 The light intensity is 100 Hz.

[0050] Additionally, the disclosed skin cosmetic methods can be used in combination with other skin cosmetic methods, for example, a skin cosmetic method (e.g., photorejuvenation) that uses red and blue light therapy devices to simultaneously reduce skin symptoms associated with microbial infections (e.g., acne) and repair epidermal tissue.

[0051] According to another aspect, the disclosure provides for the use of a compound according to any of the above, a pharma- ceutically acceptable salt or ester of said compound, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form thereof, a metabolite of each thereof, or any combination or mixture thereof, in combination with light in the treatment or diagnosis of a disease in a subject.

[0052] The light may be a single wavelength light or a mixed light.

[0053] In some embodiments, at least a portion of the wavelengths of the light are in the range of 10 nm to 1 mm, for example, one or more of the ranges of 10 nm to 380 nm, 380 to 780 nm, 780 nm to 3 μm, 3 μm to 30 μm, and 30 μm to 1 mm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 400 nm to 480 nm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 450 nm to 480 nm.

[0054] In some embodiments, the treatment is carried out by a method comprising step I and step II as defined in any one of the above.

[0055] According to another aspect, the present disclosure provides the use of a compound according to any of the above, a pharma- ceutically acceptable salt or ester of said compound, a prodrug, a stereoisomer, a hydrate, a solvate or a crystalline form thereof, a metabolite of each thereof, or any combination or mixture thereof, in combination with light in skin cosmetic treatment. In some embodiments, the skin cosmetic treatment is for non-therapeutic purposes.

[0056] The light may be a single wavelength light or a mixed light.

[0057] In some embodiments, at least a portion of the wavelengths of the light are in the range of 10 nm to 1 mm, for example, one or more of the ranges of 10 nm to 380 nm, 380 to 780 nm, 780 nm to 3 μm, 3 μm to 30 μm, and 30 μm to 1 mm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 400 nm to 480 nm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 450 nm to 480 nm.

[0058] In some embodiments, the skin cosmetization is carried out by a method comprising step I' and step II' as defined in any one of the above.

[0059] The compounds selected in the present disclosure, their pharma- ceutically acceptable salts or esters, prodrugs, stereoisomers, hydrates, solvates, or crystalline forms, their respective metabolites, or any combination or mixture thereof, can be used to treat or prevent the development of inflammatory bowel disorders in a subject. (i) the tumor and complications caused by the tumor; (ii) Diseases associated with microbial or parasitic infections; and (iii) Immune-related diseases The compounds can be used to treat or diagnose diseases that are

[0060] In some embodiments, the tumor and the tumor-related disease are selected from breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumor, primary liver cancer, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary tumor, testicular tumor, non-Hodgkin's lymphoma, bladder cancer, leukemia, gastric cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, cervical cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, colon cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, glioma, and secondary complications of these diseases (pain, infection, pericardial effusion, pleural effusion, etc.).

[0061] In some embodiments, the microorganism is Bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Shigella dysenteriae, Bacillus pertussis, Bacillus diphtheria, Diplococcus meningitidis, Mycobacterium tuberculosis, Clostridium tetanus, Bacillus leprosy, Group A hemolytic streptococci, Brucella, Bacillus cholera, Bacillus typhi, Bacillus anthrax, Neisseria gonorrhoeae, Bacillus acnes, and Salmonella paratyphi A, B, or C, etc.); Viruses (such as influenza virus, mumps virus, rubella virus, encephalitis B virus, dengue virus, epidemic hemorrhagic fever virus, rabies virus, human papilloma virus, polio virus, measles virus, varicella zoster virus, hepatitis virus, novel enterovirus 70, mutant strains of coxsackievirus A24, and human immunodeficiency virus) Fungi (such as Candida albicans, Trichophyton rubrum, and Epidermophyton floccosum), Mycoplasma (such as Mycoplasma pneumoniae, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalis), Chlamydia (Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia psittacosis, and Chlamydia of livestock, etc.) Rickettsiae (Rickettsia typhus, Rickettsia mooseri, Rickettsia rocky mountain spotted fever, and Rickettsia tsutsugamushi), Actinomycetes (such as Actinomyces israelii), or Spirochetes (Leptospira, Treponema pallidum, etc.) is selected from.

[0062] In some embodiments, the parasite is Roundworms, hookworms, tapeworms, Trichomonas vaginalis, liver fluke, Paragonimus westermani, Toxoplasma gondii, sarcoidosis, Trichinella spiralis, ameba, Leishmania donovani, malaria parasite, schistosomes, filariasis, hydatids, scabies, demodex, lice, or fleas is selected from.

[0063] In some embodiments, the disease associated with a microbial or parasitic infection is Extraintestinal infections or diarrhea caused by Escherichia coli, viral hepatitis, bacterial dysentery, amoebic dysentery, gonorrhea, syphilis, polio, measles, whooping cough, diphtheria, epidemic cerebrospinal meningitis, scarlet fever, epidemic hemorrhagic fever, rabies, leptospirosis, brucellosis, anthrax, epidemic encephalitis type B, black fever, malaria, dengue fever, tuberculosis, schistosomiasis, filariasis, echinococcosis, leprosy, influenza, mumps, rubella, neonatal tetanus, acute hemorrhagic conjunctivitis, cholera, typhoid fever caused by Salmonella typhi, paratyphoid fever, mycoplasmal pneumonia, nongonococcal urethritis, mycoplasmal cervicitis cervicitis), trachoma, psittacosis, chlamydia pneumoniae, typhus, typhus fever, Rocky Mountain spotted fever, rickettsial pox, scrub typhus, spotted fever, trench fever, ringworm (such as tinea pedis, black tinea palm, nodose trichomycosis, tinea pedis, tinea manus, tinea corporis, tinea cruris, onychomycosis, and tinea capitis), sporotrichosis, yeast infection, candidiasis, aspergillosis, cryptococcosis, zygomycosis, Penicillium Marneffei, condyloma acuminata, shingles, AIDS, pulmonary actinomycosis, acne, and other infectious diarrheal diseases. is selected from.

[0064] In some embodiments, the immune related disease is: Secondary immunodeficiencies such as infections (such as rubella, measles, leprosy, tuberculosis, cytomegalovirus infection, HIV infection, and coccidioidomycosis), protein loss (such as nephrotic syndrome and protein-losing enteropathy), immunoglobulin synthesis defects, lymphocyte loss (such as lymphocyte loss caused by drugs and / or systemic infections), and other diseases (such as diabetes, cirrhosis of the liver, and subacute sclerosing panencephalitis) and / or secondary immunodeficiencies caused by immunosuppressive therapy, and Autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism, juvenile diabetes mellitus, primary platelet purpura, autoimmune hemolytic anemia, ulcerative colitis, skin diseases, and chronic liver disease is selected from.

[0065] Additionally, the compounds selected in the present disclosure, their pharma- ceutically acceptable salts or esters, prodrugs, stereoisomers, hydrates, solvates, or crystalline forms, their respective metabolites, or any combination or mixture thereof, can be used to treat or diagnose a precancerous lesion or skin disease in a subject, or for skin cosmetics.

[0066] In some embodiments, Precancerous lesions are Precancerous lesions of the cervix caused by papillomavirus infection, and Precancerous lesions of breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumor, primary liver cancer, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary tumor, testicular tumor, non-Hodgkin's lymphoma, bladder cancer, leukemia, stomach cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, colon cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, and glioma is selected from.

[0067] In some embodiments, the skin disease is selected from genital warts, acne associated with microbial infection, port wine stains, actinic keratosis, skin cancer, precancerous lesions of skin cancer, and benign proliferative diseases of the skin.

[0068] In some embodiments, skin cosmesis includes the removal of pigmented skin spots (liver spots, spots, etc.), skin shedding, and the alleviation of acne associated with microbial infections.

[0069] The compounds selected in the present disclosure, pharma- ceutically acceptable salts or esters of the compounds, prodrugs, stereoisomers, hydrates, solvates, or crystal forms, their respective metabolites, or any combination or mixture thereof, can be made into any pharma- ceutically acceptable preparation, such as a transdermal formulation (e.g., ointments, plasters, patches, paints, sprays, gels, etc.), drops, perfusates, injections (e.g., liquid injections, injection powders, or injection tablets), oral formulations (e.g., oral solid formulations such as tablets, capsules, pills, granules, or oral liquid formulations such as oral liquids, oral suspensions, syrups, etc.).

[0070] Additionally, the compounds, pharma- ceutically acceptable salts or esters of the compounds, prodrugs, stereoisomers, hydrates, solvates or crystalline forms, their respective metabolites, or any combination or mixture thereof selected in the disclosure, can be formulated into any formulation suitable for application to the surface of the skin, such as creams, ointments, lotions, gels, and the like.

[0071] In some embodiments, the formulation comprises one or more pharma- ceutically acceptable auxiliary agents (e.g., substrates, excipients, carriers, stabilizers, or solubilizers). Pharmaceutically acceptable carriers that can be used include, but are not limited to, sterile liquids such as water and oils, including oils of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. When the formulation is administered by injection, water is used as an exemplary excipient. Saline, aqueous glucose, or aqueous glycerol solutions can also be specifically used as liquid excipients for injection. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, calcium carbonate (chalk), silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, nonfat dry milk, glycerin, propylene glycol, water, and ethanol. The formulation can further contain minor amounts of wetting agents, emulsifying agents, or pH buffering agents, if desired. Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, and magnesium carbonate. Examples of suitable pharma- ceutical acceptable excipients are described in Non-Patent Document 1. Gels can be made from natural, semi-synthetic, or synthetic polymeric materials such as alginates, gelatin, pectin, cellulose derivatives, starch and its derivatives, povidone, polyvinyl alcohol, and polyacrylic acid (e.g., carbomer, polyacrylic acid), as a matrix.

[0072] In some embodiments, the adjuvant is water or saline.

[0073] According to another aspect, the present disclosure further relates to a method for inhibiting activity of a tumor cell, a microorganism, or a parasite, the method comprising: i) applying a compound according to any of the above, a pharma- ceutically acceptable salt or ester of said compound, a prodrug, a stereoisomer, a hydrate, a solvate, or a crystalline form of said compound, a metabolite of each thereof, or any combination or mixture thereof, to a tumor cell, a microorganism, or a parasite; Step ii: Irradiating the tumor cells, microorganisms, or parasites with light; Includes.

[0074] In step ii, single wavelength light or mixed light can be used.

[0075] In some embodiments, at least a portion of the wavelengths of the light in step ii are in the range of 10 nm to 1 mm, for example, one or more of the ranges of 10 nm to 380 nm, 380 nm to 780 nm, 780 nm to 3 μm, 3 μm to 30 μm, and 30 μm to 1 mm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 400 nm to 480 nm. In some embodiments, at least a portion of the wavelengths of the light are in the range of 450 nm to 480 nm.

[0076] In some embodiments, the irradiation in step ii is performed for 1 second to 12 hours, for example, 1 second to 10 seconds, 10 seconds to 100 seconds, 100 seconds to 200 seconds, 200 seconds to 500 seconds, or 500 seconds to 900 seconds, for example, 1 minute to 10 minutes, 10 minutes to 30 minutes, or 30 minutes to 60 minutes, for example, 1 hour to 2 hours, 2 hours to 5 hours, or 5 hours to 12 hours.

[0077] In some embodiments, the irradiation in step ii is from 1 to 2000 mW / cm 2 (For example, 1 to 15 mW / cm 2 , 15mW / cm 2 ~50mW / cm 2 , 50mW / cm 2 ~100mW / cm 2 , 100mW / cm 2 ~200mW / cm 2 , 200mW / cm 2 ~300mW / cm 2 , 300mW / cm 2 ~500mW / cm 2 , 500mW / cm 2 ~1000mW / cm 2 , 1000mW / cm 2 ~1500mW / cm 2 , or 1500mW / cm2 ~2000mW / cm 2 ) of light intensity.

[0078] The method can be performed in vivo or in vitro in a subject and can be used for therapeutic or non-therapeutic purposes. In some embodiments, the method is used for non-therapeutic purposes and is performed in vitro. For example, the method can be used to inhibit the activity of microorganisms or parasites on a surface of a human or animal (excluding wounds and infected areas) or on a surface of a subject, or the method can be used in in vitro experiments for research purposes.

[0079] In some embodiments, the tumor cells are Breast cancer cells, melanoma cells, meningioma cells, soft tissue sarcoma cells, salivary gland tumor cells, primary liver cancer cells, intraspinal tumor cells, mediastinal tumor cells, brain cancer cells, bone cancer cells, penile cancer cells, osteosarcoma cells, intracranial tumor cells, tongue cancer cells, maxillary sinus cancer cells, thyroid cancer cells, malignant lymphoma cells, multiple myeloma cells, pituitary tumor cells, testicular tumor cells, non-Hodgkin's lymphoma cells, bladder cancer cells, leukemia cells, gastric cancer cells, nasopharyngeal cancer cells, laryngeal cancer cells, oral cancer cells, esophageal cancer cells, lung cancer cells, renal cancer cells, cervical cancer cells, choriocarcinoma cells, vulvar cancer cells, skin cancer cells, endometrial cancer cells, ovarian cancer cells, prostate cancer cells, pancreatic cancer cells, colon cancer cells, rectal cancer cells, colorectal cancer cells, Kaposi's sarcoma cells, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma) cells, hemangioma cells, or glioma cells is selected from.

[0080] In some embodiments, the microorganism is Bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Shigella dysenteriae, Bordetella pertussis of the Bacillus genus, Corynebacterium diphtheriae of the Bacillus genus, Neisseria meningitidis of the Diplococcus genus, Mycobacterium tuberculosis, Clostridium tetanus, Leprosy of the Bacillus genus, Group A hemolytic streptococci, Brucella, Vibrio cholerae of the Bacillus genus, Salmonella typhi of the Bacillus genus, Bacillus anthrax, Salmonella acnes, Salmonella paratyphi A, Salmonella paratyphi B, or Salmonella paratyphi C, etc.); Viruses (such as influenza virus, mumps virus, rubella virus, encephalitis type B virus, dengue fever virus, epidemic hemorrhagic fever virus, rabies virus, human papilloma virus, polio virus, measles virus, varicella zoster virus, hepatitis virus, novel enterovirus 70, mutant strains of coxsackievirus A24, and human immunodeficiency virus), Fungi (such as Candida albicans, Trichophyton rubrum, and Epidermophyton inguinale), Mycoplasma (such as Mycoplasma pneumoniae, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalis), Chlamydia (Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia psittacosis, and Chlamydia of livestock, etc.) Rickettsiae (such as Rickettsia typhus, Rickettsia typhus, Rickettsia rocky mountain spotted fever, and Rickettsia tsutsugamushi), Actinomycetes (such as Actinomyces israelii), or Spirochetes (Leptospira, Treponema pallidum, etc.) is selected from.

[0081] In some embodiments, the parasite is Roundworms, hookworms, tapeworms, Trichomonas vaginalis, liver fluke, Paragonimus westermani, Toxoplasma gondii, sarcoidosis, Trichinella spiralis, ameba, Leishmania donovani, malaria parasite, schistosomes, filariasis, hydatids, scabies, demodex, lice, or fleas is selected from.

[0082] In the present disclosure, the subject is preferably a mammal, such as a bovine, equine, porcine, canine, feline, rodent, or primate, such as a human.

[0083] [Definition of terms] In the present disclosure, the term "photodynamic therapy" refers to photochemotherapy, which is based on the activation of photosensitizers to generate reactive oxygen species (ROS) represented by singlet oxygen, and can achieve the purpose of treating diseases by killing tumor cells or pathogenic microorganisms, etc. Photodynamic therapy can also be used for skin cosmetics, such as removing wrinkles, removing sagging skin, removing pigmented skin lesions, removing skin thickening, and removing keratin.

[0084] In the present disclosure, the term "photodynamic diagnosis" refers to a photochemical diagnostic method in which a disease is diagnosed under the action of light using a characteristic spectrum of a photosensitizer that has an affinity for tumor or other diseased tissue.

[0085] In this disclosure, the term "single-wavelength light" refers to light whose spectral intensity distribution peak has a half-width of 10 nm or less.

[0086] In this disclosure, the term "mixed light" refers to light obtained by mixing multiple single wavelength lights.

[0087] In this disclosure, the term "photosensitizer" refers to a chemical that absorbs light energy of a specific wavelength and transfers the energy to other molecules, thereby producing reactive oxygen species and other substances capable of killing cells or microorganisms.

[0088] In this disclosure, the term "microorganism" includes bacteria, viruses, fungi, and some small protists, microalgae, and other small organisms that are difficult to see with the naked eye. Microorganisms that cause disease in humans or animals are referred to as "pathogenic microorganisms" and generally include fungi, actinomycetes, spirochetes, bacteria, rickettsia, chlamydia, viruses, and mycoplasma. Effect of the Invention

[0089] The present disclosure has the following advantageous effects.

[0090] The diketone photosensitizer of organic small molecule used in the present disclosure has weak intermolecular aggregation effect, low toxicity, good water solubility, and maintains good photochemical properties in the aqueous phase, and is cheap and easily available. The diketone compound used in the present disclosure has the following technical effects under photoexcitation: (1) Inhibition of tumor cell activity, (2) Inhibition of the activity of bacteria and other microorganisms; (3) Improved immunity (4) good bioavailability, and (5) Good safety Achieve one or more of the following.

[0091] The present disclosure has significant clinical value since it can be used to treat tumors or diseases caused by microbial or parasitic infections, can be used to improve immunity, and can also be used for skin cosmetics.

[0092] The embodiments of the present disclosure will be described in detail below with reference to the drawings and examples. However, those skilled in the art will understand that the following drawings and examples are only used to illustrate the present disclosure and are not used to limit the scope of the present disclosure. Various objects and advantageous aspects of the present disclosure will be apparent to those skilled in the art based on the drawings and the following detailed description of the preferred embodiments. [Brief description of the drawings]

[0093] [Figure 1]FIG. 1 shows the change in tumor tissue size in BALB / C mice during two weeks of treatment in Experimental Example 3. As shown, 14 days after administration, the difference between the high-concentration treatment group and the control group is very significant (p<0.001), and the difference between the low-concentration treatment group and the control group is also very significant (p<0.001). The tumor size is regulated and reduced, and the effect is more significant in the high-concentration treatment group. This experiment shows that diketone compounds have an inhibitory effect on tumors. [Diagram 2] FIG. 2 shows the weight change of tumor tissue in BALB / C mice after 2 weeks of treatment in Experimental Example 3. As shown, 14 days after administration of diketone compound, the difference between the high concentration treatment group and the control group is very significant (p<0.001), and the difference between the low concentration treatment group and the control group is also very significant (p<0.001). Based on the tumor weight ratio, the growth inhibition effect is evident in the treatment group, and the high concentration treatment group shows a better inhibitory effect on tumors. This experiment shows that diketone compound has an inhibitory effect on tumors. [Diagram 3] Figure 3 shows the number of B cells in tumor tissue of BALB / C mice after 2 weeks of treatment in Experimental Example 3. As shown in the figure, 14 days after administration of diketone compound, the difference between high concentration treatment group and control group is very significant (p<0.001), and the difference between low concentration treatment group and control group is also very significant (p<0.01). The number of B cells in tumor tissue is significantly increased in treatment group, and the effect of improving immunity is more favorable in high concentration treatment group. [Figure 4] FIG. 4 shows the levels of aspartate aminotransferase in the blood of BALB / C mice after two weeks of treatment in Experiment 3. [Diagram 5] FIG. 5 shows the blood urea nitrogen levels of BALB / C mice after two weeks of treatment in Experiment 3. [Figure 6] FIG. 6 shows the levels of creatine kinase in the blood of BALB / C mice after two weeks of treatment in Experiment 3. [Figure 7]FIG. 7 shows the changes in body weight of BALB / C mice during the two-week treatment in Experimental Example 4. [Figure 8] FIG. 8 shows the levels of aspartate aminotransferase in the blood of BALB / C mice after two weeks of treatment in Experiment 4. [Figure 9] FIG. 9 shows the blood urea nitrogen levels of BALB / C mice after two weeks of treatment in Experiment 4. [Figure 10] FIG. 10 shows the levels of creatine kinase in the blood of BALB / C mice after two weeks of treatment in Experiment 4. [Figure 11] FIG. 11 shows the toxicity of the experimental photosensitizer group at different concentrations against human cervical cancer Hela cells in the dark and under light in Experimental Example 5. The administered concentrations are 6×10-3mol / L, 3×10-6mol / L, 6×10-7mol / L, 6×10-9mol / L, 3×10-9mol / L, and 6×10-10mol / L. As shown, when the diketone compound is administered in the dark, the toxicity against Hela is relatively low, indicating that the diketone compound has good safety. When the diketone compound is administered under light, the toxicity against Hela is relatively high, which is significantly different from the toxicity of the diketone compound in the dark, indicating that the diketone compound has significant inhibitory activity against tumor cells under light. [Figure 12]FIG. 12 shows the toxicity of the experimental photosensitizer group to human immortalized keratinocytes (Hacat cells) in the dark and under blue light, and the toxicity of the control drug 5-α-aminolevulinic acid (ALA) group to human immortalized keratinocytes (Hacat cells) in the dark and under red light in Experimental Example 5. The administered concentrations are 6×10-3 mol / L, 6×10-4 mol / L, 6×10-5 mol / L, 6×10-6 mol / L, 6×10-7 mol / L, 6×10-8 mol / L, and 6×10-9 mol / L. As shown, ALA is more toxic to normal cells under light. The toxicity of the diketone compound to normal cells under light is significantly different from the toxicity of ALA to normal cells under light. This indicates that the diketone compound has low toxicity and high selectivity to normal cells under light. [Figure 13] FIG. 13 shows that strong fluorescence is generated on the skin of mice in the experimental photosensitizer group under light excitation at the time of administration. Moreover, 4 hours after administration, the fluorescence on the skin of the mice is significantly reduced. 8 hours after administration, the fluorescence on the skin of the mice approaches zero. The above results indicate that the selected diketone compounds of the present disclosure can be used for photodynamic therapy or diagnosis on the skin. Furthermore, the diketone compounds have good safety because they are rapidly metabolized and do not remain in or outside the body for a long time.

[0094] The embodiments of the present disclosure are described in detail below with reference to examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present disclosure, and are not used to limit the scope of the present disclosure. Those without specific conditions in the examples are generally carried out under conventional conditions or conditions recommended by the manufacturer. Reagents or equipment used without specifying the manufacturer are all conventional products that can be purchased commercially. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0095] [Bioactivity experiment] The cells, reagents, and equipment for the in vitro experiments in the following experimental examples are as follows:

[0096] Drugs BJMU-201 to BJMU-214 were all purchased from companies such as J&K Scientific Co., Ltd., Beijing Ouhe Technology Co., Ltd., or Beijing InnoChem Science & Technology Co., Ltd.

[0097] "cell" SW480 colon carcinoma cells, Hela cells, and B16 melanoma cells were all provided by ATCC cell bank.

[0098] 《Culture solution》 RPMI1640 medium and fetal bovine serum (FBS), DMEM (containing 2 mL of glutamine and Earle's BSS, 1.5 g / L NaHCO3, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate solution), and FBS were used.

[0099] 《Cell culture》 Cells were incubated at 37° C., 5% CO 2 , and saturated humidity until 80% confluence, then digested with 0.25% trypsin-EDTA.

[0100] Main related reagents and equipment: The following equipment was used: DMEM high sugar medium, 1640 medium, and trypsin (Gibco, Maryland, USA), FBS (PAN, Germany), a high-speed low-temperature refrigerated centrifuge (Beijing DLAB Scientific, China), a horizontal shaker (ZD-9550, Kylin Bell Lab Instruments, Jiangsu Province), an ultra-purification workbench (Suzhou Antai Airtech, Suzhou Province), and a multi-function microplate reader FlexStation3 (Molecular Devices). EXAMPLES

[0101] [Experimental Example 1: Evaluation of the toxicity of compounds to tumor cells] In this experimental example, the toxicity of the compounds against tumor cells was tested by the MTT method, which included the following specific steps.

[0102] (1) Logarithmic growth phase cells were harvested, the concentration of the cell suspension was adjusted, and the cell suspension was added to a cell culture plate at a volume of 100 μL per well. Then, cells were added to the cell culture plate to adjust the density of the cells to be tested to 1,000-10,000 wells (the peripheral wells were filled with sterile PBS).

[0103] (2) The cells were incubated at 37°C in the presence of 5% CO2 until the monolayer of cells covered the bottom of each well (96-well flat-bottom plate), and then a gradient of drug concentrations was added. Typically, 5-7 gradients of drugs were added at a volume of 100 μL per well in a multi-well setup of 3-5 wells.

[0104] (3) The cells were randomly divided into a light treatment group and a dark treatment group, incubated at 37°C in the presence of 5% CO2 for 16 to 48 hours, and observed under an inverted microscope.

[0105] (4) The cells were illuminated with a laser of wavelengths between 450 nm and 480 nm (110 mW / cm 2 The final dose was approximately 100 J / cm for 900 seconds. 2 After light irradiation, the cells were washed with PBS.

[0106] (5) MTT solution (5 mg / ml, 0.5% MTT) was added at a volume of 20 μL per well, and the cells were further incubated for 4 hours. If the drug could react with MTT, centrifugation was first performed, then the culture solution was discarded, and the cells were carefully washed with PBS two or three times, and then the culture solution containing MTT was added.

[0107] (6) After the incubation was completed, the incubation solution in the wells was carefully removed.

[0108] (7) Dimethyl sulfoxide was added at a volume of 150 μL per well, and then the plate was shaken on a shaker at low speed for 10 minutes to completely dissolve the crystals. The absorbance of each well at an optical density (OD) of 490 nm was measured using an enzyme-linked immunosorbent assay (ELISA).

[0109] (8) Zero adjustment wells (medium, MTT, dimethyl sulfoxide) and control wells (cells, same concentration of lysis medium, culture solution, MTT, dimethyl sulfoxide) were set up simultaneously.

[0110] Table 3 shows the toxicity of 14 test compounds (BJMU-201 to BJMU-214) against three tumor cells under light. The results show that the selected diketone compounds of the present disclosure have relatively high cytotoxicity against SW480 cells, Hela cells, and B16 cells when administered under light. The above results show that the compounds of the present disclosure have significant inhibitory activity against tumor cells.

[0111] [Table 3]

[0112] Table 4 shows the toxicity of 14 test compounds (BJMU-201 to BJMU-214) against three tumor cells in the dark. The results show that the selected diketone compounds of the present disclosure have relatively low cytotoxicity against SW480 cells, Hela cells, and B16 cells when administered in the dark. The above results show that the compounds of the present disclosure have good safety.

[0113] [Table 4] EXAMPLES

[0114] [Experimental Example 2: Evaluation of the inhibitory effect of compounds on bacteria] A single E. coli colony was transferred from a solid LB (Luria Bertani) agar plate into 5 ml of liquid LB medium and incubated at 37°C for 12 hours. The bacteria were harvested by centrifugation (7000 rpm, 1 min) and washed three times with PBS buffer. The supernatant was discarded and the remaining E. coli cells were resuspended in PBS buffer. The bacterial suspension was prepared to an optical density (OD600) of 1.0. The suspension was then diluted (5 times) with PBS buffer. The diluted E. coli suspension was incubated in the presence of a 60 mM solution of a photosensitizer (diketone compound) at 37°C for 15 minutes in the dark, and then exposed to a 15 mW / cm 2 Laser light (wavelength 450nm to 480nm, approximately 3J cm -2 After irradiation, the bacterial suspension was serially diluted (10 to 10) in PBS buffer. 4 100 μL of diluted bacterial E. coli cells were spread on a solid LB agar plate and incubated at 37°C for 12 to 16 hours, and the colonies formed were counted. Meanwhile, a treated group in the dark and an untreated group in the dark were set up, and the inhibition rate was determined by dividing the number of colony forming units (cfu) killed in the treated group under light or the treated group in the dark by the number of colony forming units (cfu) killed in the untreated group in the dark.

[0115] The results are shown in Table 5. The diketone compounds have a significant inhibitory effect on the growth of E. coli under light. The results show that the compounds of the present disclosure have significant antibacterial activity. The diketone compounds have a slight inhibitory effect on the growth of E. coli in the dark. The results show that the compounds of the present disclosure have good safety.

[0116] [Table 5] EXAMPLES

[0117] [Experimental Example 3: Evaluation of the tumor suppression effect of compounds (animal experiments)] 1. Photodynamic therapy experiments of tumors were carried out using BJMU-204 as a photosensitizer and BALB / C mice as experimental animals.

[0118] "Model Style" BALB / C mice were divided into a total of 5 groups, with 11-12 mice in each group. 4T1 cells (breast cancer cells in mice) were resuscitated and subcultured to a better cell condition for tumor formation inoculation. BALB / C mice were locally depilated and disinfected, and tumor cells were injected in situ into the mouse mammary fat pad. Tumors were formed within 7-10 days.

[0119] Administration time Drug therapy was administered to mice after tumor formation, using a laser beam with a wavelength of 450 nm to 480 nm and an intensity of 200 mW / cm. 2 Photodynamic therapy was administered intratumorally at a light intensity of 100x on alternate days.

[0120] Mode of Administration As shown in Table 6 below, the photosensitizer was dissolved in saline and injected intratumorally into the mice of groups 1 to 3, and the groups receiving photodynamic therapy were irradiated with light immediately after injection.

[0121] [Table 6]

[0122] 2. Mice in each experimental group were subjected to blood sampling for serum separation and necropsy, and tumors were removed and measured to obtain tumor wet weight. Tumor tissues were fixed and pathologically examined, and tumor changes in mice were observed under light microscope using hematoxylin-eosin staining. Routine blood indexes (white blood cell count (WBC), red blood cell count (RBC), lymphocyte count (LY), platelet count (PLT), and many other routine blood indexes) and blood biochemical indexes (including routine biochemical indexes of blood) were examined and analyzed in the laboratory of Peking University Third Hospital.

[0123] 3. The tumors were removed from the mice of each experimental group, and tumor tissue pieces of approximately 1 cm x 1 cm x 0.5 cm in size were cut and placed in 10 ml EP tubes. 5 ml of pancreatin was added for digestion. After 30 minutes, the digestion was completed. The digestion fluid and tumor tissue were sieved through a single cell sieve to obtain a single cell suspension of tumor tissue. B220 antibody (for flow cytometry) was added to the suspension, and the resulting solution was stained with PI staining solution. After 30 minutes, the staining was completed and the staining solution was removed. Then, resuspension was performed with PBS solution for testing using flow cytometry.

[0124] Figure 1 shows the change in tumor tissue size in BALB / C mice during 2 weeks of treatment. As shown, after 14 days of treatment, the difference between the high-concentration treatment group and the control group is very significant (p<0.001), and the difference between the low-concentration treatment group and the control group is also very significant (p<0.001). The tumor size is regulated and reduced, and this effect is more significant in the high-concentration treatment group. This experiment shows that diketone compounds have an inhibitory effect on tumors.

[0125] Figure 2 shows the weight change of tumor tissue in BALB / C mice after 2 weeks of treatment. As shown, after 14 days of treatment, the difference between the high-concentration treatment group and the control group is very significant (p<0.001), and the difference between the low-concentration treatment group and the control group is also very significant (p<0.001). Based on the tumor weight ratio, the growth inhibition effect is evident in the treatment group, and the high-concentration treatment group shows a better inhibition effect on tumors. This experiment shows that diketone compounds have an inhibitory effect on tumors.

[0126] Figure 3 shows the number of B cells in tumor tissue of BALB / C mice after 2 weeks of treatment. As shown, after 14 days of treatment, the difference between the high-concentration treatment group and the control group is very significant (p<0.001), and the difference between the low-concentration treatment group and the control group is very significant (p<0.01). The number of B cells in tumor tissue is significantly increased in the treatment group, and the effect of improving immunity is better in the high-concentration treatment group.

[0127] Table 7 shows the results of routine blood tests in BALB / C mice after two weeks of treatment. The platelet and white blood cell counts in the blood of the mice are reduced, and the difference between the treatment group and the control group is very significant (p<0.001). The platelet and white blood cell counts are reduced in the treatment group, indicating that the immune function of the treatment group is improved. This experiment shows that photodynamic therapy containing diketone compounds improves immunity.

[0128] Figures 4-6 show blood biochemistry data of BALB / C mice after two weeks of treatment. Figures 4, 5, and 6 correspond to the levels of aspartate aminotransferase, urea nitrogen, and creatine kinase, respectively.

[0129] The normal blood data of BALB / C mice in Table 7 (excluding LY and PLT) and the blood biochemistry data in Figures 4 to 6 show that the diketone compounds do not cause liver, kidney, and blood toxicity after 14 days of continuous treatment at high or low therapeutic doses.

[0130] [Table 7] EXAMPLES

[0131] [Experimental Example 4: Evaluation of phototoxicity of compounds in healthy mice (animal experiment)] 1. Phototoxicity evaluation experiments were carried out using BJMU-204 as a photosensitizer and BALB / C mice as experimental animals.

[0132] BALB / C mice were divided into two groups, 9 mice per group. The mice were adaptively fed for 2 days. The mice in the control group were normally fed, and the mice in the experimental group were irradiated with 200 mW / cm 2 Photodynamic therapy was performed by subcutaneously injecting laser light with a wavelength of 450 to 480 nm at a light intensity of 100 nm every other day.

[0133] Administration Form As shown in Table 8 below, the photosensitizer was dissolved in saline and injected subcutaneously into the mice in the experimental group, and the group receiving photodynamic therapy was irradiated with light immediately after injection.

[0134] [Table 8]

[0135] 2. The mice in the experimental and control groups were weighed every other day. At the end of the experiment, the mice were subjected to blood sampling for serum separation and autopsy, and the blood routine indexes (including WBC, RBC, LY, PLT, and many other normal blood indexes) and blood biochemistry indexes (including normal blood biochemistry indexes) were examined and analyzed in the laboratory of Peking University Third Hospital.

[0136] Table 9 shows the routine blood data for BALB / C mice.

[0137] [Table 9]

[0138] Figure 7 shows the change in body weight of BALB / C mice during 2 weeks of treatment. As shown, 14 days after administration, the difference in body weight between the control group and the experimental group is not significant (p<0.05), and there is no significant change in the body weight of the experimental group during the treatment period. This experiment shows that diketone compounds have little phototoxicity to normal mice.

[0139] Figures 8-10 show blood biochemistry data of BALB / C mice after two weeks of treatment. Figures 8, 9, and 10 correspond to the levels of aspartate aminotransferase, urea nitrogen, and creatine kinase, respectively.

[0140] The normal blood data of BALB / C mice in Table 9 and the blood biochemistry data in Figures 8 to 10 show that the diketone compounds do not cause liver, kidney, and blood toxicity after being treated in healthy mice at high therapeutic doses for 14 consecutive days. EXAMPLES

[0141] [Experimental Example 5 - Evaluation of the cytotoxicity of compounds] In this experimental example, the toxicity of compound BJMU-204 to cells was tested by MTT method, which includes the following specific steps.

[0142] (1) Cells in the logarithmic growth phase were harvested, the concentration of the cell suspension was adjusted, and the cell suspension was added to a cell culture plate at a volume of 100 μL per well. Then, cells were added to the cell culture plate to adjust the density of the cells to be tested to 1,000 to 10,000 wells (the surrounding wells were filled with sterile PBS).

[0143] (2) The cells were incubated at 37°C in the presence of 5% CO2 until the monolayer of cells covered the bottom of each well (96-well flat-bottom plate), and then a gradient of drug concentrations was added. Typically, 5-7 gradients of drugs were added at a volume of 100 μL per well, with 3-5 wells set up for each drug concentration.

[0144] (3) The cells were randomly divided into light-treated and dark-treated groups, then incubated at 37°C for 16-48 hours in the presence of 5% CO2, and observed under an inverted microscope. The cells in the experimental photosensitizer-light-treated group were incubated for 15 minutes and then irradiated with light. The cells in the control 5-α-aminolevulinic acid (ALA)-light-treated group were incubated for 3 hours and then irradiated with light (the control drug ALA produced porphyrin for light absorption only after 3 hours of incubation). MTT solution was added to the mice in the experimental photosensitizer-non-irradiated group immediately after 15 minutes of incubation, or to the mice in the control ALA-non-irradiated group after 3 hours of incubation.

[0145] (4) For the experimental photosensitizer-light irradiation group, cells were exposed to a laser with a wavelength of 450 nm to 480 nm (intensity 110 mW / cm 2 ) for 900 seconds, resulting in a final dose of approximately 100 J / cm. After light exposure, the cells were washed with PBS.

[0146] In the control ALA-illumination group, cells were exposed to light with wavelengths of 630 to 650 nm (intensity 110 mW / cm 2 ) laser for 900 seconds, with a final dose of approximately 100 J / cm. After photoexposure, the cells were washed with PBS.

[0147] (5) MTT solution (5 mg / ml, 0.5% MTT) was added at a volume of 20 μL per well, and the cells were further incubated for 4 hours. If the drug could react with MTT, centrifugation was first performed, then the culture solution was discarded, and the cells were carefully washed with PBS two or three times, and then the culture solution containing MTT was added.

[0148] (6) After the incubation was completed, the incubation solution in the wells was carefully removed.

[0149] (7) Dimethyl sulfoxide was added at a volume of 150 μL per well, and then the plate was shaken at low speed on a shaker for 10 minutes to completely dissolve the crystals. The absorbance of each well at OD490 nm was measured by enzyme-linked immunosorbent assay.

[0150] (8) Zero adjustment wells (medium, MTT, dimethyl sulfoxide) and control wells (cells, same concentration of lysis medium, culture solution, MTT, dimethyl sulfoxide) were set up simultaneously.

[0151] Figure 11 shows the toxicity of the experimental photosensitizers against human cervical cancer HeLa cells in the dark and under light. The dose concentrations were 6×10 -3 mol / L, 3×10 -6 mol / L, 6×10 -7 mol / L, 6×10 -9 mol / L, 3×10 -9 mol / L and 6×10 -10 mol / L. The results show that the selected diketone compounds of the present disclosure have relatively low cytotoxicity to human cervical cancer Hela cells when administered in the dark. The above results show that the compounds of the present disclosure have good safety. The results show that the selected diketone compounds of the present disclosure have relatively high cytotoxicity to human cervical cancer Hela cells when administered under blue light. The above results show that the compounds of the present disclosure have significant inhibitory activity against tumor cells under light.

[0152] Figure 12 shows the toxicity of the experimental photosensitizers to human immortalized keratinocytes (Hacat cells) in the dark and under blue light, and the toxicity of the control drug ALA to human immortalized keratinocytes (Hacat cells) in the dark and under red light. The dose concentration was 6 x 10 -3 mol / L, 6×10 -4 mol / L, 6×10 -5 mol / L, 6×10 -6 mol / L, 6×10 -7 mol / L, 6×10 -8mol / L and 6×10 -9 mol / L. The results show that the selected diketone compounds of the present disclosure have relatively low cytotoxicity to human immortalized keratinocytes (Hacat cells) when administered in the dark and under blue light. The control drug ALA has relatively high cytotoxicity to human immortalized keratinocytes (Hacat cells) when administered under red light. The toxicity of the selected diketone compounds of the present disclosure to normal cells under light is significantly different from the toxicity of ALA to normal cells under light. The above results show that the compounds of the present disclosure have low toxicity to normal cells and high selectivity under light. EXAMPLES

[0153] [Experimental Example 6 - Evaluation of compound fluorescence in mouse skin] This example evaluates the potential of compound BJMU-204 as a photosensitizer administered to the skin. The specific steps are as follows:

[0154] BALB / C mice were depilated, the skin was thoroughly washed, and then the mice were protected from light for more than 4 hours. Photographs of the backs of the mice were taken using a small animal live imager IVIS SPECTRUM, with excitation at wavelengths of 480 nm and 520 nm for 2 seconds. Mice that did not receive photosensitizers were photographed as controls. Mice that received 100 μL of 30% compound on the skin of their backs (10 mm diameter) were photographed, and the fluorescence intensity was recorded, and then photographed 2, 4, 8, and 12 hours after compound administration. Each image was processed using Living Image 4.3.1, and the average fluorescence in a circular field of 10 mm diameter on the skin was measured. The ambient light and background fluorescence of the mouse skin were corrected based on the fluorescence value of the backs of mice that did not receive the compound.

[0155] FIG. 13 shows that strong fluorescence is produced on the skin of mice in the experimental photosensitizer group under the excitation of light at the time of administration. The results indicate that the selected diketone compounds of the present disclosure can be used for photodynamic therapy or photodynamic diagnosis on the skin. The diketone compounds emit fluorescence quickly after being excited by light, and there is no need to wait for a long time. Furthermore, 4 hours after administration, the fluorescence on the skin of the mice is significantly reduced. 8 hours after administration, the fluorescence on the skin of the mice is approaching zero. The above results indicate that the compounds of the present disclosure are metabolized quickly and do not remain in the body for a long time, and therefore have good safety.

[0156] The above experiments show that the selected diketone compounds of the present disclosure exhibit significant killing activity against tumor cells under light, and significant inhibitory activity against bacterial growth under light, and in the dark, the diketone compounds do not exhibit these two activities and have low toxicity. The selected diketone compounds of the present disclosure further significantly inhibit the activity of tumor cells in mice and significantly improve immune activity. In summary, the selected diketone compounds of the present disclosure can be used in photodynamic therapy to achieve the purpose of treating cancer, microbial infection, and related complications, and can also be used to treat immune-related diseases.

[0157] Although specific embodiments of the present disclosure have been described in detail, those skilled in the art will understand that, in accordance with all the teachings disclosed, various modifications and substitutions can be made to those details, all of which are within the scope of the present disclosure. The full scope of the present disclosure is provided by the appended claims and any equivalents thereof.

Claims

1. In the preparation of a medicament or reagent, 【Chemistry 1】 or a pharma- ceutically acceptable salt or ester, prodrug, stereoisomer, hydrate, solvate, or crystalline form of said compound, or a metabolite of each of them, or any combination or mixture thereof, wherein said medicament or said reagent is used in the photodynamic therapy or photodynamic diagnosis of a disease or for skin cosmetology by photodynamic therapy, In the formula (I), R １ and R ２ are each independently linear or branched alkyl having 1 to 6 carbon atoms; halogenated straight or branched alkyl having 1 to 6 carbon atoms; a hydroxyl group, or Aryl, 5-6 membered heteroaryl, 5-6 membered heterocyclyl, and 3-6 membered cycloalkyl functional groups having 6-14 carbon atoms, optionally substituted with one or more (e.g., 2, 3, 4, or 5) identical or different substituents; and The substituents are each independently selected from the group consisting of a hydroxyl group, a carboxyl group, a sulfone group, a halogen (e.g., fluorine, chlorine, bromine, or iodine) atom, an amino group, a mercapto group, a nitro group, -C(O)- (straight or branched alkyl having 1 to 4 carbon atoms), -S(O) ２ -O-(linear or branched alkyl having 1 to 4 carbon atoms) or -O-(linear or branched alkyl having 1 to 4 carbon atoms); Preferably, the linear or branched alkyl having 1 to 6 carbon atoms is a linear or branched alkyl having 1 to 4 carbon atoms; Preferably, the linear or branched alkyl having 1 to 4 carbon atoms is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, or t-butyl; Preferably, the aryl having 6 to 14 carbon atoms is phenyl or naphthyl; Preferably, the halogenated linear or branched alkyl having 1 to 6 carbon atoms is a fluorinated, chlorinated, brominated or iodinated linear or branched alkyl having 1 to 6 carbon atoms; Preferably, the 5-6 membered heteroaryl contains 1 to 3 ring atoms selected from nitrogen, oxygen, and sulfur; Preferably, the 5- to 6-membered heterocyclyl contains 1 to 3 ring atoms selected from nitrogen, oxygen, and sulfur. use.

2. The R １ is selected from linear or branched alkyl having 1 to 4 carbon atoms (e.g., methyl or ethyl), or aryl having 6 to 14 carbon atoms (e.g., phenyl); The R ２ is selected from a linear or branched alkyl having 1 to 4 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl, butyl, or t-butyl), a halogenated linear or branched alkyl having 1 to 4 carbon atoms (e.g., bromomethyl or bromoethyl), or a hydroxyl group.

2. The use according to claim 1.

3. The compound is (1) R １ is a straight or branched alkyl having 1 to 4 carbon atoms, and R ２ is a straight or branched chain alkyl having 1 to 4 carbon atoms; (2) R １ is a straight or branched alkyl group having 1 to 4 carbon atoms or a phenyl group, and R ２ is a hydroxyl group, (3) R １ is a straight or branched alkyl having 1 to 4 carbon atoms, and R ２ is a halogenated straight or branched chain alkyl having 1 to 4 carbon atoms; (4) R １ and R ２ each is phenyl, and said phenyl is optionally substituted with one or more substituents, which may be the same or different; or (5) R １ and R ２ each independently represents a nitrogen-containing 5- to 6-membered heteroaryl, and the nitrogen-containing 5- to 6-membered heteroaryl is optionally substituted with one or more identical or different substituents; Characterized in 3. Use according to claim 1 or 2.

4. The compound is 【Table 1】 And, 【Table 2】 Selected from Use according to any one of claims 1 to 3.

5. The photodynamic therapy comprises: Step I of administering to a subject the compound of any one of claims 1 to 4, a pharma- ceutically acceptable salt or ester, prodrug, stereoisomer, hydrate, solvate, or crystalline form of said compound, or a metabolite of each thereof, or any combination or mixture thereof; A step II of irradiating the target with light; The method includes the steps of: Preferably, the method comprises the steps of: (1) The light in step II is single wavelength light or mixed light; (2) at least a portion of the wavelengths of the light in step II is in the range of 10 nm to 1 mm; (3) the irradiation in step II is performed for 1 second to 12 hours; and (4) The irradiation in step II is 1 to 2000 mW / cm ２ Doing so with a light density of Characterized in one or more of the following: Use according to any one of claims 1 to 4.

6. The disease is (i) tumors and complications due to such tumors; (ii) diseases associated with microbial or parasitic infections, and (iii) immune-related disease is selected from Preferably, the tumor and its complications are: Breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumor, primary liver cancer, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary tumor, testicular tumor, non-Hodgkin's lymphoma, bladder cancer, leukemia, gastric cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, cervical cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, colon cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, glioma, and secondary complications of these diseases (pain, infection, pericardial effusion, pleural effusion, etc.) is selected from Preferably, the microorganism is Bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Shigella dysenteriae, Bordetella pertussis of the genus Bacillus, Corynebacterium diphtheriae of the genus Bacillus, Neisseria meningitidis of the genus Diplococcus, Mycobacterium tuberculosis, Clostridium tetanus, Bacillus leprosy, Group A hemolytic streptococcus, Brucella, Vibrio cholerae of the genus Bacillus, Salmonella typhi of the genus Bacillus, Bacillus anthrax, Neisseria gonorrhoeae, Salmonella acnes, Salmonella paratyphi A, Salmonella paratyphi B, or Salmonella paratyphi C, etc.); Viruses (such as influenza virus, mumps virus, rubella virus, encephalitis type B virus, dengue fever virus, epidemic hemorrhagic fever virus, rabies virus, human papilloma virus, polio virus, measles virus, varicella zoster virus, hepatitis virus, novel enterovirus 70, mutant strains of coxsackievirus A24, and human immunodeficiency virus); Fungi (such as Candida albicans, Trichophyton rubrum, and Epidermophyton inguinale), Mycoplasma (such as Mycoplasma pneumoniae, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalis), Chlamydia (Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia psittacosis, Chlamydia of livestock, etc.), Rickettsiae (such as Rickettsia typhus, Rickettsia typhus, Rickettsia rocky mountain spotted fever, and Rickettsia tsutsugamushi) Actinomycetes (such as Actinomyces israelii), or spirochetes (such as Leptospira and Treponema pallidum) and Preferably, the parasite is Roundworms, hookworms, tapeworms, Trichomonas vaginalis, Clonorchis sinensis, Paragonimus westermani, Toxoplasma gondii, cysticercosis porcina, Trichinella spiralis, ameba, Leishmania donovani, Plasmodium falciparum, Schistosoma blood flukes, filariasis, hydatids, scabies, demodicids, lice, and fleas is selected from Preferably, the disease associated with infection with said microorganism or said parasite is Extraintestinal infections or diarrhea caused by Escherichia coli, viral hepatitis, bacterial dysentery, amoebic dysentery, gonorrhea, syphilis, polio, measles, whooping cough, diphtheria, epidemic cerebrospinal meningitis, scarlet fever, epidemic hemorrhagic fever, rabies, leptospirosis, brucellosis, anthrax, epidemic encephalitis type B, black fever, malaria, dengue fever, tuberculosis, schistosomiasis, filariasis, echinococcosis, leprosy, influenza, mumps, rubella, neonatal tetanus, acute hemorrhagic conjunctivitis, cholera, typhoid fever caused by Salmonella typhi, paratyphoid fever, mycoplasma pneumonia , nongonococcal urethritis, mycoplasmal cervicitis, trachoma, psittacosis, pneumoniae chlamydia, typhus, typhus fever, Rocky Mountain spotted fever, rickettsial pox, scrub typhus, spotted fever, trench fever, ringworm (such as tinea pedis, tinea manus, tinea corporis, tinea cruris, onychomycosis, and tinea capitis), sporotrichosis, chromophytosis, candidiasis, aspergillosis, cryptococcosis, zygomycosis, penicillium marneffeii, condyloma acuminata, herpes zoster, AIDS, pulmonary actinomycosis, acne, and other infectious diarrheal diseases. is selected from Preferably, the immune-related disease is Secondary immunodeficiencies such as infectious diseases (such as rubella, measles, leprosy, tuberculosis, cytomegalovirus infection, HIV infection, and coccidioidomycosis), protein loss (such as nephrotic syndrome and protein-losing enteropathy), immunoglobulin synthesis defects, lymphocyte loss (such as lymphocyte loss caused by drugs and / or systemic infections), and other diseases (such as diabetes, cirrhosis of the liver, and subacute sclerosing panencephalitis), and / or secondary immunodeficiencies caused by immunosuppressive therapy, and Autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, scleroderma, hyperthyroidism, juvenile diabetes mellitus, primary thrombocytopenia, autoimmune hemolytic anemia, ulcerative colitis, skin diseases, and chronic liver disease Selected from Use according to any one of claims 1 to 5.

7. the disease is a precancerous lesion or a skin disease, Preferably, the precancerous lesion is: Precancerous lesions of the cervix caused by papillomavirus infection, and Precancerous lesions of breast cancer, melanoma, meningioma, soft tissue sarcoma, salivary gland tumor, primary liver cancer, intraspinal tumor, mediastinal tumor, brain cancer, bone cancer, penile cancer, osteosarcoma, intracranial tumor, tongue cancer, maxillary sinus cancer, thyroid cancer, malignant lymphoma, multiple myeloma, pituitary adenoma, testicular tumor, non-Hodgkin's lymphoma, bladder cancer, leukemia, stomach cancer, nasopharyngeal cancer, laryngeal cancer, oral cancer, esophageal cancer, lung cancer, kidney cancer, choriocarcinoma, vulvar cancer, skin cancer, endometrial cancer, ovarian cancer, prostate cancer, pancreatic cancer, colon cancer, rectal cancer, colorectal cancer, Kaposi's sarcoma, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma), hemangioma, and glioma is selected from Preferably, the skin disease is selected from genital warts, acne associated with microbial infection, port wine stains, actinic keratosis, skin cancer, precancerous lesions of skin cancer, and benign proliferative diseases of the skin. Use according to any one of claims 1 to 5.

8. 1. A method for cosmetically treating a subject's skin, comprising: The method comprises using the compound according to any one of claims 1 to 4, a pharma- ceutically acceptable salt or ester, prodrug, stereoisomer, hydrate, solvate, or crystalline form of said compound, a metabolite of each thereof, or any combination or mixture thereof, as a photosensitizer, The skin cosmetic treatment is for non-therapeutic purposes, Preferably, the skin cosmetic treatment is the removal of pigmented spots (liver spots, age spots, etc.) on the skin, peeling, and alleviation of acne associated with microbial infection; Preferably, the method for skin cosmetic treatment is used in combination with another method for skin cosmetic treatment (such as a method for skin cosmetic treatment using red and blue light treatment devices, such as photorejuvenation). method.

9. Use of the compound according to any one of claims 1 to 4, a pharma- ceutically acceptable salt or ester, prodrug, stereoisomer, hydrate, solvate, or crystalline form of said compound, or a metabolite of each thereof, or any combination or mixture thereof, in combination with light in skin cosmetology, comprising: The skin cosmetic treatment is for non-therapeutic purposes. use.

10. 1. A non-therapeutic in vitro method for inhibiting the activity of a tumor cell, a microorganism, or a parasite, comprising: step i) applying the compound according to any one of claims 1 to 4, a pharma- ceutically acceptable salt or ester, prodrug, stereoisomer, hydrate, solvate, or crystalline form of said compound, or a metabolite of each thereof, or any combination or mixture thereof, to said tumor cells, said microorganisms, or said parasites; ii) irradiating the tumor cells, the microorganisms, or the parasites with light; The method includes:

11. (1) the light in step ii is single wavelength light or mixed light; (2) at least a portion of the wavelengths of the light in step ii) is in the range of 10 nm to 1 mm; (3) the irradiation in step ii is performed for 1 second to 12 hours; and (4) The irradiation in step ii is 1 to 2000 mW / cm ２ Doing so with a light density of Characterized in one or more of the following: The method of claim 10.

12. The tumor cells are Breast cancer cells, melanoma cells, meningioma cells, soft tissue sarcoma cells, salivary gland tumor cells, primary liver cancer cells, intraspinal tumor cells, mediastinal tumor cells, brain cancer cells, bone cancer cells, penile cancer cells, osteosarcoma cells, intracranial tumor cells, tongue cancer cells, maxillary sinus cancer cells, thyroid cancer cells, malignant lymphoma cells, multiple myeloma cells, pituitary tumor cells, testicular tumor cells, non-Hodgkin's lymphoma cells, bladder cancer cells, leukemia cells, gastric cancer cells, nasopharyngeal cancer cells, laryngeal cancer cells, oral cancer cells, esophageal cancer cells, lung cancer cells, renal cancer cells, cervical cancer cells, choriocarcinoma cells, vulvar cancer cells, skin cancer cells, endometrial cancer cells, ovarian cancer cells, prostate cancer cells, pancreatic cancer cells, colon cancer cells, rectal cancer cells, colorectal cancer cells, Kaposi's sarcoma cells, non-melanoma skin cancer (including squamous cell carcinoma and basal cell carcinoma) cells, hemangioma cells, or glioma cells and The microorganism is Bacteria (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Shigella dysenteriae, Bordetella pertussis of the genus Bacillus, Corynebacterium diphtheriae of the genus Bacillus, Neisseria meningitidis of the genus Diplococcus, Mycobacterium tuberculosis, Clostridium tetanus, Leprosy of the genus Bacillus, Group A hemolytic streptococcus, Brucella, Vibrio cholerae of the genus Bacillus, Salmonella typhi of the genus Bacillus, Bacillus anthrax, Salmonella acnes, Salmonella paratyphi A, Salmonella paratyphi B, or Salmonella paratyphi C, etc.); Viruses (such as influenza virus, mumps virus, rubella virus, encephalitis type B virus, dengue fever virus, epidemic hemorrhagic fever virus, rabies virus, human papilloma virus, polio virus, measles virus, varicella zoster virus, hepatitis virus, novel enterovirus 70, mutant strains of coxsackievirus A24, and human immunodeficiency virus); Fungi (such as Candida albicans, Trichophyton rubrum, and Epidermophyton inguinale), Mycoplasma (such as Mycoplasma pneumoniae, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalis), Chlamydia (such as Chlamydia trachomatis, Chlamydia pneumoniae, Chlamydia psittacosis, and Chlamydia of livestock), Rickettsiae (such as Rickettsia typhus, Rickettsia typhus, Rickettsia rocky mountain spotted fever, and Rickettsia tsutsugamushi) Actinomycetes (such as Actinomyces israelii), or Spirochetes (Leptospira, Treponema pallidum, etc.) and The parasite is Roundworms, hookworms, tapeworms, Trichomonas vaginalis, Clonorchis sinensis, Paragonimus westermani, Toxoplasma gondii, cysticercosis porcina, Trichinella spiralis, ameba, Leishmania donovani, Plasmodium falciparum, Schistosoma blood flukes, filariasis, hydatids, scabies, demodicids, lice, and fleas Selected from 12. The method according to claim 10 or 11.

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