Mitochondrial targeting photosensitizer, preparation method and application thereof, and medicine box containing mitochondrial targeting photosensitizer

Abstract

The present invention relates to a mitochondrial targeting photosensitizer or a pharmaceutically acceptable salt thereof wherein the mitochondrial targeting photosensitizer has a structure of the following general formula (I): # imgabs0 # wherein groups R are each independently defined in the specification. The invention further relates to preparation and application of the mitochondrial targeting photosensitizer and a medicine box containing the mitochondrial targeting photosensitizer.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and in particular relates to a mitochondrial targeted photosensitizer, a preparation method and use thereof, and a drug kit containing the same. Background Art

[0002] Targeted drugs refer to drugs or their preparations that are endowed with targeting capabilities. The purpose is to enable the drug or its carrier to target specific lesions and accumulate or release active ingredients at the target site. Targeted preparations can make the drug form a relatively high concentration at the target site, thereby improving the efficacy while inhibiting toxic side effects and reducing damage to normal tissues and cells. According to different targeting mechanisms, drug targeting can be divided into passive targeting, active targeting and physical targeting. Among them, physical targeting uses physical signals such as light, heat, magnetic field, electric field, and ultrasound to artificially regulate the distribution and release characteristics of drugs in the body to achieve targeting of lesions. Photosensitizers are targeted drugs that work by physical targeting and are used in photodynamic therapy (PDT) to effectively treat a variety of diseases. Photodynamic therapy is a new type of therapy for treating diseases. The principle is a photochemical reaction, and its basic elements are oxygen, photosensitizer and visible light. First, the target site selectively absorbs the photosensitizer, and then after local irradiation with light of appropriate wavelength, the photosensitizer is activated to produce a photosensitizing effect.

[0003] Sodium porphyrin for injection, which is currently in clinical use, is a new type of photosensitizer. It is a single effective chemical component isolated from PHOTOFRIN (porphyrin sodium). Preclinical studies on esophageal cancer have shown that the mechanism of action of sodium porphyrin for injection is the same as that of PHOTOFRIN, but to achieve the same intensity of anti-tumor activity, its dosage is only 10% of PHOTOFRIN, and the phototoxic reaction is low, which has obvious advantages in effectiveness, safety and quality control. However, it has been found in previous studies that sodium porphyrin can selectively accumulate in tumor tissues, but the drug will be distributed throughout the body, and normal tissues will also have a certain degree of accumulation. Patients need to avoid light for about 7 days after treatment. In order to reduce the phototoxicity of sodium porphyrin, it is still necessary to improve the targeting accuracy of sodium porphyrin. This study designed a mitochondrial-targeted photosensitizer to enhance the PDT effect in a hypoxic environment, thereby obtaining a photosensitizer with high activity and low toxicity. Summary of the invention

[0004] In view of the state of the prior art, the present invention aims to provide a photosensitizer that can significantly improve the targeting of tumors, thereby improving the efficacy and having lower toxicity, and the preparation method thereof has high yield and simple synthesis process. The mitochondrial photosensitizer of the present invention can be excited under light of different wavelengths and doses, and has high biological safety, and is a mitochondrial photosensitizer with potential clinical application value and can be mass-produced.

[0005] In one aspect, the present invention provides a mitochondrial targeted photosensitizer or a pharmaceutically acceptable salt thereof, wherein the mitochondrial targeted photosensitizer has the following structure of general formula (I): , Wherein the groups R each independently represent C 3-9 Heterocyclyl-O- or C 3-9 Heterocyclyl-C 1-10 Alkylene-O-, Wherein C 3-9 The heterocyclyl group is unsubstituted or substituted by one or more identical or different groups selected from the group consisting of cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, alkylcarbonyl, dialkylamino, haloalkyl or haloalkoxy; The C 1-10 The alkylene group is linear or branched, unsubstituted or substituted by one or more identical or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, alkylcarbonyl, dialkylamino, haloalkyl or haloalkoxy, and the alkylene group may be optionally interrupted by one or more heteroatoms selected from S, N and O or by one or more 3-6 membered nitrogen-containing heterocyclic groups; The 3-6 membered nitrogen-containing heterocyclic ring is unsubstituted or substituted by one or more identical or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, haloalkyl or haloalkoxy.

[0006] On the other hand, the present invention provides a method for preparing a mitochondrial targeted photosensitizer of formula (I), comprising the following steps: reacting porphyrinic acid with a G-OH compound in the presence of a catalyst and a condensing agent in the presence of a solvent to obtain a product; The GO part in the G-OH compound is the same as the definition of the R group above.

[0007] In another aspect, the present invention provides a use of a mitochondrial targeted photosensitizer or a pharmaceutically acceptable salt thereof for preparing a medicament for treating the following diseases: cancer, including reproductive organ cancer (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer and testicular cancer), leukemia, mastocytoma, breast cancer, kidney cancer, respiratory tract cancer (such as lung cancer, especially small cell and non-small cell lung cancer, as well as bronchial cancer and pleuropulmonary blastoma), brain cancer, digestive tract cancer (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, gastric cancer, pancreatic cancer, rectal cancer, small intestine cancer and salivary gland cancer), urinary tract cancer, liver cancer, eye cancer (such as intraocular melanoma and retinoblastoma), skin cancer, head and neck cancer (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer and oral cancer and squamous cell carcinoma), multiple myeloma, sarcoma or lymphoma; and precancerous lesions of the above diseases.

[0008] In another aspect, the present invention provides a method for treating the following diseases, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof; wherein the disease is selected from cancer, including reproductive organ cancer (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer and testicular cancer), leukemia, mastocytoma, breast cancer, renal cancer, respiratory tract cancer (such as lung cancer, in particular small cell and non-small cell lung cancer, and bronchial cancers of the digestive tract (such as esophagus, anus, colon, colorectum, gallbladder, stomach, pancreas, rectum, small intestine, and salivary glands), urinary tract, liver, eye (such as intraocular melanoma and retinoblastoma), skin, head and neck (such as thyroid, parathyroid, larynx, hypopharynx, nasopharynx, oropharynx, lip and oral cavity, and squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and their precursors.

[0009] In another aspect, the present invention provides a drug kit comprising a therapeutically effective amount of a mitochondrial targeting photosensitizer of formula (I) of the present invention or a pharmaceutically acceptable salt thereof; and instructions for using the same in photodynamic therapy.

[0010] Unexpectedly, the inventors found that the compound of formula (I) of the present invention is a photosensitizer that can significantly improve tumor targeting, thereby improving drug efficacy and having lower toxicity. The preparation method thereof has a high yield and a simple synthesis process, and can be used as a high-efficiency photosensitizer with potential clinical application value and can be mass-produced. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Scatter plot showing fluorescence imaging and colocalization analysis of DVDMS and MTDR.

[0012] Figure 2 The scatter plot is the fluorescence imaging and co-localization analysis of compound X-1 and MTDR.

[0013] Figure 3 Figure 2 shows the body weight changes of A549 tumor-bearing mice after drug administration.

[0014] Figure 4 The figure shows the changes in tumor volume of A549 tumor-bearing mice after drug administration.

[0015] Figure 5 The tumor weights of A549 tumor-bearing mice at each experimental endpoint. DETAILED DESCRIPTION

[0016] Hereinafter, the present invention will be described in more detail.

[0017] Unless otherwise indicated, "compound of formula (I) of the present invention", "compound of formula (I)", "compound of the present invention" and "mitochondrial targeted photosensitizer of the present invention (photosensitizer for short)" are used synonymously.

[0018] As used herein, the term "comprising" and its synonyms "including" and "containing" mean "including but not limited to", and are not intended to exclude, for example, other additives, components, integers or steps.

[0019] Unless defined differently, the names of chemical groups are generally to be understood so that the bond to the backbone or the rest of the molecule is via the last mentioned structural element of the relevant chemical group, ie, for example in the case of heterocyclylalkylene, via a carbon atom of the alkylene.

[0020] The terms "optional", "optional" or "optionally" as used herein mean that the subsequently described event, circumstance or material may or may not appear or exist, and that such description includes instances where the event, circumstance or material appears or exists and instances where the event, circumstance or material does not appear or exist.

[0021] Unless otherwise stated, the following definitions apply to radicals or substituents used throughout the present description and claims. Within the scope of the present invention, the meanings of all radicals occurring repeatedly are independent of one another.

[0022] As used herein, the term "cyano" refers to a functional group in which a carbon atom and a nitrogen atom are connected by a triple bond.

[0023] As used herein, the term "halogen" denotes, for example, fluorine, chlorine, bromine or iodine. If the term is applied to a radical, "halogen" denotes, for example, a fluorine, chlorine, bromine or iodine atom.

[0024] As used herein, the term "alkyl" refers to a saturated straight-chain or branched hydrocarbon radical having the specified number of carbon atoms in each case, for example (C1-C 10)-alkyl, (C1-C6)-alkyl and (C1-C4)-alkyl, examples of which are not limited to methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl, 2,2-dimethylbutyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, 1,1,2-trimethylpropyl, 1,2,2-trimethylpropyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, n-heptyl, 1-methylhexyl, 2-methylhexyl, 1,1-dimethylpentyl, 1-ethylpentyl, 2-ethylpentyl, 1-propylbutyl, n-octyl, 1-methylheptyl, 2-ethylhexyl, 1,3-dimethylhexyl and 1-ethyl-2-methylpentyl.

[0025] As used herein, the term "haloalkyl" refers to an alkyl group as defined above in which one or more hydrogen atoms are replaced by one or more halogen atoms, which may be the same or different. 10 )-haloalkyl, (C1-C6)-haloalkyl and (C1-C4)-haloalkyl, examples include but are not limited to chloromethyl, bromomethyl, dichloromethyl, trichloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chlorofluoromethyl, dichlorofluoromethyl, chlorodifluoromethyl, 1-chloroethyl, 1-bromoethyl, 1-fluoroethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 2-chloro-2-fluoroethyl, 2-chloro-2,2-difluoroethyl, 2,2-dichloro-2-fluoroethyl, 2,2,2-trichloroethyl, pentafluoroethyl and 1,1,1-trifluoropropan-2-yl. Preferred are fluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, pentafluoroethyl and 1,1,1-trifluoropropan-2-yl.

[0026] According to the invention, "cycloalkyl" - on its own or as part of a chemical group - represents a monocyclic, bicyclic or tricyclic hydrocarbon preferably having 3 to 10 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl or adamantyl. Also preferred are cycloalkyl having 3, 4, 5, 6 or 7 carbon atoms, such as, in particular, cyclopropyl or cyclobutyl. The cycloalkyl radicals according to the invention may be substituted by one or more identical or different radicals.

[0027] According to the invention, "alkoxy" represents a straight-chain or branched alkyl-O- preferably having 1 to 6 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy. Alkoxy groups having 1 to 4 carbon atoms are also preferred. The alkoxy groups of the invention may be substituted by one or more identical or different groups.

[0028] As used herein, the term "haloalkoxy" refers to an alkoxy group as defined above in which one or more hydrogen atoms of the alkyl portion are replaced by one or more halogen atoms, which may be the same or different. 10 )-haloalkoxy, (C1-C6)-haloalkoxy and (C1-C4)-haloalkoxy, examples include but are not limited to chloromethoxy, bromomethoxy, dichloromethoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, chlorofluoromethoxy, 1-chloroethoxy, 1-bromoethoxy, 1-fluoroethoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, 2-chloro-2-fluoroethoxy, pentafluoroethoxy. Preferred are fluoromethoxy, difluoromethoxy, trifluoromethoxy, 1-fluoroethoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, and pentafluoroethoxy.

[0029] According to the invention, "alkylthio" represents a straight-chain or branched alkyl-S- group preferably having 1 to 6 carbon atoms, such as methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, isobutylthio, sec-butylthio and tert-butylthio. Alkylthio groups having 1 to 4 carbon atoms are also preferred. The alkylthio groups of the invention may be substituted by one or more identical or different groups.

[0030] According to the invention, "alkylcarbonyl" represents a straight-chain or branched alkyl-C(=O)- preferably having 2 to 7 carbon atoms, for example methylcarbonyl, ethylcarbonyl, n-propylcarbonyl, isopropylcarbonyl, sec-butylcarbonyl and tert-butylcarbonyl. Alkylcarbonyl having 1 to 4 carbon atoms is also preferred. The alkylcarbonyl of the invention may be substituted by one or more identical or different groups.

[0031] According to the invention, "heterocyclyl" represents a carbocyclic ring system having at least one ring in which at least one carbon atom is replaced by a heteroatom, preferably by a heteroatom selected from N, O, S, P, B, Si, Se, and which is saturated, unsaturated or heteroaromatic and may be unsubstituted or substituted, wherein the bonding site is on a ring atom. Unless otherwise defined, a heterocyclic ring contains 3 to 9 ring atoms (i.e. C 3-9 Heterocyclyl), especially 3 to 6 ring atoms (ie C 3-6The heterocyclic ring is preferably a cycloheterocyclic ring, and one or more, preferably 1 to 4, in particular 1, 2 or 3 heteroatoms in the heterocyclic ring are preferably selected from N, O and S, but two oxygen atoms should not be directly adjacent. The heterocyclic ring generally contains not more than 4 nitrogen atoms and / or not more than 2 oxygen atoms and / or not more than 2 sulfur atoms. In the case of optionally substituted heterocyclic groups, the invention also includes polycyclic ring systems, for example 8-azabicyclo[3.2.1]octyl, 1-azabicyclo[2.2.1]heptyl, 1-oxa-5-azaspiro[2.3]hexyl or 2,3-dihydro-1 H -Indole.

[0032] The heterocyclic group of the present invention is, for example, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetrazinyl, oxazinyl, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, dioxane, pyrrolinyl, pyrrolidinyl, imidazoline alkyl, oxaziridinyl, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxaziridine, oxopyrrolidyl, dioxopyrrolidyl, oxomorpholinyl, oxopiperazinyl, and oxepanyl.

[0033] According to the present invention, "C 3-9 Heterocyclic group" and "C 3-9 Heterocyclyl C 1-10 The "heterocyclic group" in "alkylene" has the meaning represented above; 1-10 The alkylene group in “alkylene” means methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene and decylene, and isomers thereof.

[0034] According to the present invention, the "3-6 membered nitrogen-containing heterocyclic group" is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl or morpholinyl; preferably pyrazolyl, imidazolyl, triazolyl (for example, triazole, 1,2,4-triazole, 1,2,5-triazole, 1,3,4-triazole), oxazolyl, oxadiazolyl or furazanyl.

[0035] Any description of a compound of formula (I) anywhere herein also encompasses any diastereomers or enantiomers and E / Z isomers of the compound of formula (I), as well as salts thereof, that exist.

[0036] Depending on the nature of the substituents, the compounds of formula (I) described at any point herein may also be in the form of stereoisomers, i.e. geometric and / or optical isomers or in the form of isomer mixtures of varying composition. The invention provides both the pure stereoisomers and any desired mixtures of these isomers, although only compounds of formula (I) are generally discussed here.

[0037] If appropriate, the compounds of the formula (I) may be present in various polymorphic forms or as mixtures of various polymorphic forms. Both the pure polymorphs and the polymorph mixtures are provided by the invention and can be used according to the invention.

[0038] Compounds obtained from combinations which are contrary to the laws of nature and which the skilled person would therefore exclude based on his / her expert knowledge are not covered herein. For example, ring structures with three or more adjacent oxygen atoms are excluded.

[0039] According to one aspect of the present invention, the present invention provides a mitochondrial targeting photosensitizer or a pharmaceutically acceptable salt thereof, wherein the mitochondrial targeting photosensitizer has the following structure of general formula (I): , Wherein the groups R each independently represent C 3-9 Heterocyclyl-O- or C 3-9 Heterocyclyl C 1-10 Alkylene-O-, Wherein C 3-9 The heterocyclyl group is unsubstituted or substituted by one or more identical or different groups selected from the group consisting of cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, alkylcarbonyl, dialkylamino, haloalkyl or haloalkoxy; The C 1-10 The alkylene group is linear or branched, unsubstituted or substituted by one or more identical or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, alkylcarbonyl, dialkylamino, haloalkyl or haloalkoxy, and the alkylene group may be optionally interrupted by one or more heteroatoms selected from S, N and O or by one or more 3-6 membered nitrogen-containing heterocyclic groups; The 3-6 membered nitrogen-containing heterocyclic ring is unsubstituted or substituted by one or more identical or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, haloalkyl or haloalkoxy.

[0040] The pharmaceutically acceptable salts of the compounds of formula (I) of the present invention refer to those salts that are considered safe and suitable for use in pharmaceutical preparations. These salts are usually prepared by reacting the compounds of formula (I) with an acid or a base in order to improve the solubility, stability and bioavailability of the drug.

[0041] The pharmaceutically acceptable salt of the compound of the present invention is, for example, the product obtained by reacting the compound of formula (I) with an acid. Preferably, the acid includes, but is not limited to, hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, methanesulfonic acid, salicylic acid, trifluoroacetic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, fumaric acid, citric acid, tartaric acid, succinic acid, malic acid or glutamic acid. The non-toxic pharmaceutical base addition salt of the compound of formula (I) of the present invention includes salts of bases such as sodium, potassium, calcium, ammonium, etc., and those skilled in the art will recognize a variety of non-toxic pharmaceutically acceptable addition salts.

[0042] In a preferred embodiment of the present invention, in formula (I), each radical R has the same meaning.

[0043] In a preferred embodiment of the present invention, wherein in formula (I), said C 3-9 The heterocyclic group is selected from the group consisting of pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetrazinyl, oxazinyl, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, dioxanyl, pyrrolinyl, pyrrolidinyl, imidazolinyl , imidazolidinyl, thiazolidinyl, oxazolidinyl, dioxolanyl, dioxolenyl, pyrazolidinyl, tetrahydrofuranyl, dihydrofuranyl, oxetanyl, oxirane, azetidinyl, aziridine, oxazetidinyl, oxaziridine, oxaziridine, oxazepanyl, oxazihepanyl, azepanyl, oxopyrrolidinyl, dioxopyrrolidinyl, oxomorpholinyl, oxopiperazinyl, and oxepanyl.

[0044] In a preferred embodiment of the present invention, wherein in formula (I), said C 1-10 Alkyl groups are straight chain and unsubstituted.

[0045] In a preferred embodiment of the present invention, wherein in formula (I), said C 3-9The heterocyclic group contains at least one nitrogen atom selected from the group consisting of pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl, azetidinyl, aziridine, oxazetidinyl, oxaziridine, oxaziridine, oxepanyl, oxazihexaninyl, azepanyl, oxopyrrolidinyl, dioxopyrrolidinyl, oxomorpholinyl, and oxopiperazinyl.

[0046] In a preferred embodiment of the present invention, in formula (I), the group R is C 3-6 Heterocyclyl C 1-6 Alkylene-O-, where C 3-6 The heterocyclic group is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl, azetidinyl, aziridine, oxazetidinyl, oxaziridine, oxaziridine, oxopyrrolidinyl, dioxopyrrolidinyl, oxomorpholinyl, and oxopiperazinyl, preferably piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl; C 1-6 The alkylene group is selected from methylene, ethylene, propylene, n-butylene, n-pentylene or n-hexylene.

[0047] In a preferred embodiment of the present invention, in formula (I), the group R is C 3-6 Heterocyclyl C 1-4 Alkylene-O-, where C 3-6 The heterocyclic group is selected from the group consisting of pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, oxazinyl, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl, azetidinyl, oxazetidinyl, oxaziridine, oxaziridine, oxomorpholinyl and oxopiperazinyl, preferably morpholinyl, oxazinyl, isoxazinyl, oxazetidinyl, oxaziridine and oxaziridine; 1-4The alkylene group is selected from methylene, ethylene, propylene or n-butylene. In a preferred embodiment of the present invention, in formula (I), the group R is, for example, morpholinyl-N-ethyl.

[0048] In a preferred embodiment of the present invention, the 3-6 membered nitrogen-containing heterocyclic group is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl or morpholinyl; preferably pyrazolyl, imidazolyl, triazolyl (e.g., triazole, 1,2,4-triazole, 1,2,5-triazole, 1,3,4-triazole), oxazolyl, oxadiazolyl or furazanyl.

[0049] In a preferred embodiment of the present invention, wherein in formula (I), the mitochondrial targeted photosensitizer or a pharmaceutically acceptable salt thereof is used to treat the following diseases: cancer, including reproductive organ cancer (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer and testicular cancer), leukemia, mastocytoma, breast cancer, kidney cancer, respiratory tract cancer (such as lung cancer, especially small cell and non-small cell lung cancer, as well as bronchial cancer and pleural pulmonary blasts), cancers of the digestive tract (such as esophagus, anus, colon, colorectum, gallbladder, stomach, pancreas, rectum, small intestine, and salivary glands), urinary tract, liver, eye (such as intraocular melanoma and retinoblastoma), skin, head and neck (such as thyroid, parathyroid, larynx, hypopharynx, nasopharynx, oropharynx, lip and oral cavity, and squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and their precursors.

[0050] The term "precancerous lesions" in this article refers to abnormal or excessive cell proliferation, which has a high potential for canceration.

[0051] The definitions of radicals listed above in general terms or in preferred ranges can be combined with one another as desired, ie including combinations between the preferred ranges given.

[0052] Very particular preference is given to the following compounds of the formula (I) according to the invention,

[0053] The R group is morpholinylethylene-O-.

[0054] According to another aspect of the present invention, the present invention also provides a method for preparing a compound of formula (I), comprising the following steps: In the presence of a catalyst and a condensing agent, porphyrinic acid is reacted with a G-OH compound in the presence of a solvent to obtain a product; The GO part in the G-OH compound is the same as the definition of the R group above.

[0055] In the method of the present invention, the catalyst used is an organic base catalyst selected from N,N-diisopropylethylamine (DIPEA), N,N-dimethylcyclohexylamine, N,N-dimethylbenzylamine, N-methylmorpholine, N-ethylmorpholine, N-methylimidazole (NMI), 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, pyridine, N,N-dimethylaminopyridine, 2,6-lutidine or a mixture thereof, more preferably N-methylimidazole; the organic solvent used is selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide, formamide, dimethyl sulfoxide, acetone, pyridine or a mixture thereof, more preferably N,N-dimethylformamide.

[0056] In the method of the present invention, the condensing agent used is a urea cation type condensing agent, which is selected from O-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TATU), O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (TATU), borate (TBTU), O-(1,2-dihydro-2-oxy-pyridyl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU); carbodiimide type condensing agent, such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1,3-dicyclohexylcarbodiimide (DCC); or carbonyl imidazole type condensing agent, such as N,N-carbonyldiimidazole (CDI); or a mixture thereof, preferably selected from HATU, EDCI, CDI or a mixture thereof.

[0057] In the method of the present invention, the molar ratio of the catalyst to the condensation agent is (0.7-2.0):1, preferably (0.8-1.8):1, and more preferably (0.9-1.5):1.

[0058] The method for preparing the compound of formula (I) of the present invention uses a combination of a specific organic base catalyst and a specific condensing agent, especially a specific ratio of N-methylimidazole and HATU, and can obtain a target compound with high purity in good yield through simple post-treatment. If the amount of the catalyst or the condensing agent is too low or too high, it will have an adverse effect on the target product, such as an increase in the proportion of by-products and a low yield of the target product.

[0059] In the method of the present invention, the molar ratio of porphyrinic acid to the R-OH compound is 1:(4.0-10.0), preferably 1:(5.0-8.0), and more preferably 1:(6.0-7.0).

[0060] The molar ratio of the porphyrinic acid to the catalyst is 1:(4.0-12.0), preferably 1:(6.0-10.0), and more preferably 1:(8.0-9.0).

[0061] In the method of the present invention, the condensation reaction time is 0.5-24 hours, preferably 1-20 hours, more preferably 1-18 hours.

[0062] Another aspect of the present invention provides the use of the mitochondrial targeted photosensitizer or a pharmaceutically acceptable salt thereof for preparing a medicament for treating the following diseases: cancer, including reproductive organ cancer (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer and testicular cancer), leukemia, mastocytoma, breast cancer, kidney cancer, respiratory tract cancer (such as lung cancer, especially small cell and non-small cell lung cancer, as well as bronchial cancer and pleuropulmonary blastoma), brain cancer, digestive tract cancer (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, gastric cancer, pancreatic cancer, rectal cancer, small intestine cancer and salivary gland cancer), urinary tract cancer, liver cancer, eye cancer (such as intraocular melanoma and retinoblastoma), skin cancer, head and neck cancer (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer and oral cancer and squamous cell carcinoma), multiple myeloma, sarcoma or lymphoma; and hyperproliferative diseases of the above diseases, such as precancerous lesions.

[0063] Another aspect of the present invention provides a method for treating the following diseases, comprising administering to a subject in need thereof a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt thereof; wherein the disease to be treated is selected from: cancer, including reproductive organ cancer (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer and testicular cancer), leukemia, mastocytoma, breast cancer, renal cancer, respiratory tract cancer (such as lung cancer, especially small cell and non-small cell lung cancer, and bronchial cancer and pleuropulmonary blastoma), brain cancer, digestive tract cancer (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small intestine cancer, and salivary gland cancer), urinary tract cancer, liver cancer, eye cancer (such as intraocular melanoma and retinoblastoma), skin cancer, head and neck cancer (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, and oral cavity cancer, and squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and hyperproliferative conditions of the above conditions, such as precursors to cancer.

[0064] As used herein, the term "therapeutically effective amount" refers to that amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response that is being sought or desired by a researcher, physician or other clinician in a tissue, system, animal, individual or human.

[0065] As used herein, the term "treating" means killing, inhibiting or slowing the growth or increase in size of a hyperproliferative cell mass or group or a tumor or cancerous growth, reducing the number of hyperproliferative cells, or preventing spread to other anatomical sites, as well as methods of reducing the size of a hyperproliferative growth or the number of hyperproliferative cells. However, it should be understood that "treating" does not necessarily mean curing or completely eliminating a hyperproliferative growth.

[0066] Another aspect of the present invention provides a drug kit, which comprises a therapeutically effective amount of the mitochondrial targeting photosensitizer or a pharmaceutically acceptable salt thereof; and instructions for using the drug in photodynamic therapy.

[0067] As used herein, the term "kit" means any commercial package, which comprises a container for holding a compound of the present invention or a pharmaceutical preparation comprising the same, and optionally further comprises a separate container such as a separate vial or a separate foil package, for example, to hold a reconstituted dissolved matrix. The container may be any conventional shape or form known in the art, which is made of a pharmaceutically acceptable material.

[0068] In a preferred embodiment, the present invention provides a medicine kit, which comprises a lyophilized preparation of a therapeutically effective amount of a compound of the present invention for treating the diseases as described above; a dissolving matrix (such as water for injection containing auxiliary materials such as cosolvents) for reconstituting the lyophilized preparation for administration; and instructions for using the compound of the present invention as a photosensitizer in photodynamic therapy.

[0069] The various components of the kit, such as the compound of the present invention or a pharmaceutically acceptable salt thereof, a pharmaceutical preparation containing the same, a dissolving matrix, other active ingredients for treating cancer or precancerous lesions, etc., can be packaged in separate containers. Regardless of the number or type of containers, the kit can also include a device for assisting in administering the drug to the patient. The device can be a patch, an inhaler, a syringe, a pipette, a spoon with a measuring unit, or any delivery device approved for medical use.

[0070] The photosensitizer of the present invention can be administered in any of the following ways: orally, orally, by oral mucosal administration, by spray inhalation, by rectal administration, by nasal administration, by vaginal administration, by topical administration, by parenteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial injection or infusion, or by administration via an explanted reservoir, wherein topical administration, intraperitoneal or intravenous administration are preferred.

[0071] The photosensitizer of the present invention can be administered in a unit dosage form. The dosage form can be a liquid dosage form, a semisolid dosage form and a solid dosage form. The liquid dosage form can be a true solution, a colloid, a microparticle dosage form, a suspension dosage form. The semisolid dosage form can be an ointment, a cream, a paste, a gel, etc. Other dosage forms can be, for example, tablets, capsules, dripping pills, aerosols, pills, powders, solutions, emulsions, granules, suppositories, freeze-dried powder injections, inclusion compounds, implants, patches, liniments, and sustained-release agents.

[0072] For oral preparations, oral tablets and capsules can be prepared by methods known in pharmacy, and tablets can also be coated; oral liquids can be prepared as water-oil suspensions, solutions, emulsions, syrups, or dry products, which can be supplemented with water or other suitable media before use. Flavoring agents or colorants can be added if necessary.

[0073] For extragastric administration, the liquid dosage form is usually prepared from the drug of the present invention and a sterile carrier. The preferred carrier is water. Depending on the selected carrier and the drug concentration, the drug can be dissolved in the carrier or made into a suspension solution. When making a solution for injection, the drug is first dissolved in water, filtered and sterilized, and then filled into a sealed bottle or ampoule.

[0074] It can also be made into sterile preparations for injection, including crystalline powder injection, freeze-dried powder injection, etc.

[0075] When applied topically to the skin or mucous membrane, the drug of the present invention can be prepared in the form of an appropriate ointment, lotion, gel or paste, wherein the active ingredient is suspended or dissolved in one or more carriers. Alternatively, it can be prepared in the form of a microneedle or the like for use with the aid of an instrument.

[0076] Photosensitizer of the present invention also comprises pharmaceutically acceptable carrier, excipient and / or other auxiliary agent.When comprising pharmaceutically acceptable carrier, excipient and / or other auxiliary agent, usually effective dose of photosensitizer of the present invention or its pharmaceutically acceptable salt and one or more pharmaceutically acceptable carrier, excipient and / or other auxiliary agent are combined to make suitable application form or dosage form, and this procedure comprises that component is mixed, granulated, compressed, dissolved or lyophilized by suitable method.The content of carrier in medicine can be 1 to 98 weight %.For convenience, other auxiliary agents such as local anesthetic, preservative, buffer can be directly dissolved in carrier.

[0077] The photosensitizer of the present invention can be prepared into an injection, a topical preparation or an oral preparation by methods known to those skilled in the art. The photosensitizer of the present invention is preferably an injection, which is injected intravenously.

[0078] The pharmaceutically acceptable carriers, excipients and / or other auxiliary agents that can be used to prepare the photosensitizer of the present invention or its administration form are all conventional pharmaceutically acceptable carriers, excipients and / or other auxiliary agents known to those skilled in the art for this purpose.

[0079] The optimal dosage and interval of the photosensitizer of the present invention are determined by the properties of the compound and external conditions such as the form, route and site of administration, the specific mammal to be treated, the wavelength of light used for treatment, the light power and irradiation time, the type and severity of the disease to be treated, etc. This optimal dosage can be determined by conventional techniques. The optimal course of treatment, i.e., the daily dosage of the compound of the present invention or the drug of the present invention within a specified time, can be determined by methods known in the art.

[0080] The photosensitizer of the present invention is used at a dosage of 0.01-100.0 mg / kg of the subject, for example, 0.01-50.0 mg / kg of the subject, preferably 0.05-10.0 mg / kg of the subject, particularly 0.1-6.0 mg / kg of the subject, and 0.2-5.0 mg / kg of the subject; and laser illumination at a wavelength of 300-800nm, preferably 600nm-650nm, particularly 630nm is used, and the irradiation is usually performed once; the light dose is 1-300J, preferably 20-200J, and more preferably 50-150J; the optical power of the laser as the light source is usually 50-800mW, preferably 100-500mW; the illumination time is 10-2400 seconds, preferably 60-1800 seconds, and more preferably 100-1500 seconds; the illumination delay is usually 4-48 hours, preferably 10-25 hours.

[0081] In the context of the present invention, the term "optical power" refers to the actual optical power at the light irradiation site, which is measured by an optical power meter to determine the actual power at the treatment site.

[0082] In the context of the present invention, the term "light dose" refers to the actual light dose at the light irradiation site, which is obtained by multiplying the light power by the irradiation time.

[0083] In the present invention, unless otherwise explicitly stated, the contents and percentages in the context of this application are all based on weight; unless otherwise explicitly stated, the method steps of the present invention are carried out at normal temperature and pressure. Unless otherwise explicitly stated, the reagents used in the following synthetic examples are all commercially available conventional products.

[0084] In the method for preparing the compound of formula (I) of the present invention, the reaction progress is monitored by normal phase silica gel thin layer chromatography (Thin Layer Chromatography, TLC), and the reaction is judged to be complete by observing the gradual disappearance of the raw material spots. The detection is performed on a thin layer chromatograph. The thin layer chromatograph that can be used in the method of the present invention is a commercially available conventional thin layer chromatograph, such as a three-purpose ultraviolet analyzer purchased from Shanghai Heqi Glass Instrument Co., Ltd., model: WHF-204B.

[0085] In the above method for preparing the compound of the present invention, the method may optionally include other post-treatment steps. The post-treatment steps may include conventional purification steps such as adjusting the pH value, crystallization, extraction, filtration, concentration under reduced pressure, and drying. Each of the above steps can be carried out in a conventional manner known to those skilled in the art. If present, extraction is usually carried out using a mixed solution of dichloromethane and methanol, preferably a mixed solution of dichloromethane / methanol = 10:1 (v / v); drying is usually carried out by freeze drying, infrared drying, vacuum drying, etc., preferably freeze drying.

[0086] In a preferred embodiment of the present invention, the method may further include a purification step by a chromatographic column. The purification may be performed by a normal phase silica gel column, the silica gel particle size is 30-100 μm, preferably 40-63 μm, the loading amount is 20-120 g, preferably 40 g, and elution is performed using dichloromethane / methanol (v / v) (elution gradient 100% / 0% to 90% / 10%, gradient elution time 15 min).

[0087] Unless explicitly stated otherwise, all operations were performed at room temperature; reagents used were either commercially available or prepared by methods known to those skilled in the art.

[0088] The compounds of the present invention can be prepared according to the above methods. However, it should be understood that a person skilled in the art, based on his own general knowledge and available publications, can adjust the methods according to the specific circumstances of the respective compounds of the present invention that he wishes to synthesize.

[0089] Detailed synthesis examples of selected compounds of the present invention are given below. However, these examples are merely illustrative and should not be interpreted in any way as limiting the scope of the present invention.

[0090] The NMR peaks reported in the synthetic examples are 1 H NMR spectral data were obtained on a Bruker 400 MHz NMR spectrometer and the signals listed have the meanings given below: s = singlet, d = doublet, dd = doublet of doublets, m = multiplet, br s = broad singlet. The deuterated solvents used in each case are also listed in 1 H NMR spectral data are specified.

[0091] In the present invention, in addition to the above-mentioned NMR peak list 1 In addition to the H NMR spectral data, the structures of the compounds prepared in the synthetic examples were also characterized by liquid chromatography-mass spectrometry (LC-MS). The LC-MS data were obtained using a Waters instrument (model: SQD2). Synthesis Example

[0092] Table 1 Synthetic reagents

[0093] Table 2: Synthesis equipment

[0094] Synthesis of compound X-1

[0095]

[0096] At room temperature (20°C), porphyrinic acid (50 mg, 0.044 mmol, 1.0 equ) was dissolved in 5 mL DMF, and then HATU (133 mg, 0.350 mmol, 8.0 equ) and N-methylimidazole (29 mg, 0.350 mmol, 8.0 equ) were added in sequence at 20°C. After reacting at room temperature for 1 hour, N-(2-hydroxyethyl)morpholine (34 mg, 0.263 mmol, 6.0 equ) was added and reacted at room temperature for 12 hours. The reaction was detected by normal phase thin layer chromatography until the reaction of the raw material was complete. The reaction solution was poured into 100 mL of water and extracted three times with a mixed solution of dichloromethane / methanol (10 / 1, V / V) (50 mL × 3). The organic phases were combined and then washed with 200 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by a normal phase silica gel column with a particle size of 40-63 μm and a loading of 40 g (dichloromethane / methanol (V / V) 100% / 0% to 90% / 10%, 15 min) to obtain the target compound X-1 (61 mg, yield 87%).

[0097] LC-MS (m / z): 1597.1 [M+H] + 1 H NMR (400 MHz, DMSO- d 6 ): δ 10.36 (s, 2H), 10.25 (s, 2H), 9.90 (s, 2H),9.29 (br s, 2H), 8.48 (dd, J = 18.0, 11.6 Hz, 2H), 6.44(d, J= 18.0Hz, 2H)),6.22-6.18(m, 4H), 4.46–4.40 (m, 4H), 3.93–3.87 (m, 4H), 3.83 (s, 6H), 3.73(s, 6H), 3.24-3.19 (m, 4H), 3.10-3.08 (m, 12H), 2.94-2.90 (m, 10H), 2.82-2.72(m, 4H), 2.34-2.30(m, 7H), 2.26-2.21(m, 4H), 2.09-2.06(m, 4H), 2.03-1.95 (m,14H), 1.82-1.80 (m, 8H), 1.74 (s, 3H), -3.97 (br s, 4H).

[0098] Targeting capability evaluation Human lung cancer cell line A549 was used as the research object to detect the subcellular localization (mitochondrial targeting) and distribution of mitochondrial porphyrin sodium (DVDMS) and targeted photosensitizer X-1 in tumor cells.

[0099] 1. Materials and Equipment Table 3 Materials

[0100] Table 4 Equipment List

[0101] 2. Experimental system Cell information: A549 cells (Cat. No.: FH0045) were purchased from Fuheng Biotechnology.

[0102] Cell culture: Take out the cells from the -80℃ freezer and revive them, and culture them at 37℃ until they grow well. The cells are grown in RPMI-1640 complete medium containing 10% fetal bovine serum, 1% penicillin (10,000 U / mL)-streptomycin (10000 μg / mL), in a CO2 constant temperature incubator at 37℃, saturated humidity, and 5% CO2.

[0103] 3. Experimental Methods Table 5 Drug preparation table

[0104] 3.2 Cell administration and staining DVDMS and X-1 stock solutions (4 mg / mL) were prepared into DVDMS and X-1 working solutions (4 μg / mL) respectively using RPMI-1640 medium for later use.

[0105] After cell digestion and resuspension, the cells were diluted to 1.5 × 10 5 Add 1 mL of cell suspension to a glass-bottomed culture dish and culture at 37°C for more than 20 hours to allow the cells to adhere.

[0106] Remove the old culture medium with a pipette, add 1 mL of 4 μg / mL DVDMS working solution and X-1 working solution to two culture dishes respectively, and incubate at 37°C in the dark for 4 hours.

[0107] After the incubation, remove the old culture medium and add 1 mL of serum-free and phenol red-free RPMI-1640 culture medium. Repeat the operation twice.

[0108] Prepare MTDR working solution.

[0109] Aspirate the old medium, add the diluted 250 nM MTDR working solution, and incubate the staining at 37ºC for 15 minutes.

[0110] Aspirate the old culture medium and add 1 mL of serum-free and phenol red-free RPMI-1640 culture medium. Repeat the operation twice.

[0111] Aspirate the old culture medium, add 1 mL of universal tissue fixative (neutral), and fix the cells at room temperature for 10 minutes.

[0112] Aspirate the fixative solution, add 1 mL of PBS, and repeat the operation twice.

[0113] 3.3 Imaging cells using fluorescence microscopy Use bright field to find the cells, adjust the objective lens to 40x, and adjust the focus to make the mitochondria image clear.

[0114] Use a 640 nm light source for excitation and a 690 nm fluorescence channel for receiving, observe the fluorescence of MTDR, and take images.

[0115] Use 405 nm light source for excitation and 630 nm for receiving fluorescence channel to observe the fluorescence of DVDMS and X-1 and take images.

[0116] 3.4 Experimental Results After exciting DVDMS or X-1 with a 405 nm light source, good fluorescence imaging can be observed in the 630 nm receiving channel. The imaging results and co-localization analysis scatter plots of the two are shown in Figure 2. Figure 1 and Figure 2 shown.

[0117] like Figure 1The mitochondrial fluorescent probe MTDR clearly marked the structure of mitochondria in the cell. DVDMS was mainly enriched around the nucleus, where the distribution of mitochondria was also relatively concentrated. However, the colocalization of DVDMS and MTDR was weak, with a Pearson correlation coefficient of 0.376, indicating that its mitochondrial targeting was poor.

[0118] like Figure 2 , X-1 and MTDR have good co-localization, both of them can clearly show the structure of mitochondria, and the Pearson correlation coefficient is 0.901. It can be considered that X-1 has good mitochondrial targeting.

[0119] Depend on Figure 1 and 2 As shown, compared with DVDMS, X-1 has better mitochondrial targeting, which is expected to improve its ability to carry out targeted photodynamic killing of tumors in vivo, improve efficacy and reduce toxic side effects.

[0120] 4.1 Experimental design Table 6 Experimental design

[0121] Note: Dosing volume: 10 µL / g based on mouse body weight 4.2 Materials and Equipment Table 7 Materials

[0122] Table 8 Equipment List

[0123] 4.3 Experimental animals and feeding management Experimental Animals: Strain: Balb / c-nude mice; Age: 6-8 weeks old; Weight: 18-20 g; Gender: female; Quantity: 24; Supplier: Shanghai Model Organisms Technology Co., Ltd. Animal production license number: SCXK (Shanghai) 2019-0002 Animal use license number: SYXK (Shanghai) 2018-0002 Feeding and management: Animals were kept in IVC (independent ventilation system) cages (5 per cage) in the SPF animal room of Qishang Biological. The animal information card of each cage indicated the number of animals in the cage, gender, strain, receipt date, dosing regimen, experiment number, group and start date of the experiment. All cages, bedding and drinking water were sterilized before use. Cages, feed and drinking water were changed once a week. The breeding environment and lighting conditions were as follows: Temperature: 20~26℃ Humidity: 30~70% Photoperiod: 12 hours light, 12 hours no light Cage: Made of polycarbonate. Bedding is corn cobs, changed once a week.

[0124] Food: The experimental animals had free access to food (irradiated sterilized, dry pelleted food) throughout the experimental period.

[0125] Drinking water: Experimental animals were free to drink sterile water.

[0126] Cage identification: The animal information card for each cage should indicate the number of animals in the cage, gender, strain, receipt date, dosing regimen, experiment number, group and start date of the experiment.

[0127] Animal identification: ear cropping.

[0128] 4.4 Experimental methods 1. Cell Culture A549 cells were cultured in vitro in F12K medium with 10% fetal bovine serum and cultured in a 5% CO2 cell culture incubator at 37ºC. Routine treatment and subculture were performed twice a week. When the cell saturation reached 80%-90% and the number reached the required level, cells were harvested and counted, and the cell density was adjusted to 5×10 using PBS. 7 cells / ml for future use.

[0129] 2. Animal Inoculation Animals were acclimated for 7 days after arrival in the animal room. Experiments were conducted after the acclimation period. 0.1 mL (5.0×10 6 A549 cells / mouse were subcutaneously inoculated on the right back of each mouse near the thigh.

[0130] 3. Daily observation of experimental animals The use and welfare of experimental animals were carried out in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). The health status and mortality of animals were monitored daily, and routine examinations included observing the effects of tumor growth and drug treatment on the daily behavior of animals, such as behavioral activities, food and water intake (only visual observation), physical signs or other abnormal conditions.

[0131] 4. Compound Preparation Table 9 Compound preparation table

[0132] 4.5. Randomization After cell inoculation, tumor growth was observed regularly. When the tumor grew to an average tumor volume of 158 mm on day 21, 3 At 14:00, 24 animals with appropriate tumor size were randomly divided into 4 groups according to the tumor size, with 6 animals in each group. The drug was administered on the day of grouping, which was recorded as D0. Except for the erlotinib group, which was administered by gavage every day, the other groups were administered once by tail vein. The specific dosing schedule is shown in Table 6. The drug solution of each group was prepared and used before the experiment and refrigerated in the dark before use. Except for the tumor illumination link, the tumor-bearing animals were strictly protected from light for 3 days after tail vein administration, and then fed in weak light.

[0133] 4.6. Lighting The G3-G4 group received light therapy once 19 h after drug administration. Using a 630 nm semiconductor laser, the optical fiber with a microlens at the end was pointed directly at the target site, so that the beam was projected vertically and completely covered the tumor. The parameters were set according to the following steps. The mice were anesthetized by inhalation of isoflurane using an anesthesia machine for small animals. After anesthesia, they were fixed with medical tape to fully expose the tumor site, and the non-tumor site was covered with a black plastic bag.

[0134] a. Turn on the laser therapy device for calibration and set parameters.

[0135] b. When measuring the laser treatment distance, the laser spot diameter should be able to cover the size of the tumor.

[0136] c. At the laser treatment distance given in the previous step, use an optical power meter to measure the laser power per unit area.

[0137] d. Adjust the laser power value to 318 mW.

[0138] e. Set the laser treatment time and perform PDT treatment on experimental animals.

[0139] Table 10 Light dose design table

[0140] 4.7. Data Collection After group administration, the length and width of the tumor were measured twice a week with a vernier caliper (purchased from Sanfeng Precision Measuring Instruments (Shanghai) Co., Ltd.), and the mice were weighed at the same time. Tumor volume (TV) and tumor growth inhibition rate (TGI) were calculated. 体积 (%), relative tumor inhibition rate (T / C%). The specific calculation formula is as follows:

[0141] V0 is the average tumor volume of a treatment group at the beginning of drug administration, V t It is the average tumor volume of the treatment group at the end of drug administration.

[0142] At the end of the experiment, mice in each group were euthanized, and tumor tissues were removed and weighed. 重量 Percentage and tumor growth inhibition rate TGI 重量 (%). The specific calculation formula is as follows:

[0143] Among them, TW 给药组 represents the tumor weight of the drug-treated group, TW 阴性对照组 represents the tumor weight of the negative control group.

[0144] 4.8. Sample collection and processing At the end of the experiment, mice in each group were euthanized, and the tumor tissues were removed and weighed.

[0145] Data processing and statistical analysis All experimental results are expressed as "mean ± standard error". The number of days after grouping was used as the horizontal axis, and the weight or tumor volume of the mice was used as the vertical axis. GraphPad Prism 8 was used to draw the graph. Statistical analysis was performed based on the data obtained at the end of the experiment to evaluate the differences between the groups. Ordinary one-way ANOVA in GraphPad Prism 8 software was used for data analysis, and p<0.05 was considered to be significantly different.

[0146] 5. Results Weight changes During the experiment, the mice tolerated the experiment well, and the weight changes of mice in each group were shown in Figure 3 (Data points represent mean body weight within group).

[0147] Tumor volume The changes in the average tumor volume of the A549 non-small cell lung cancer subcutaneous transplant tumor model over time are shown in Table 11.

[0148] Table 11 Tumor volume of each group at different time points

[0149] Note: a. Mean ± standard error; b. Days after the start of drug administration Tumor growth curve and endpoint tumor weight Tumor growth curves and tumor weights Figure 4 and Figure 5 Data points represent the mean body weight within the group, and error bars represent the standard error (SEM).

[0150] Anti-tumor efficacy evaluation index The growth inhibition rate of the test substance on the A549 non-small cell lung cancer subcutaneous transplant tumor model was calculated based on the tumor volume on day 22 after administration (Table 12). The tumor weights of all groups on day 22 are shown in Table 13.

[0151] Table 12 Growth inhibition rate of the test substance on A549 non-small cell lung cancer subcutaneous transplanted tumor model

[0152] Note: a. Mean ± standard error; b. p The values ​​were analyzed by one-way ANOVA Table 13 Tumor weight and statistical analysis at the end of the experiment (day 22)

[0153] Note: a. Mean ± standard error; b. p Values ​​were analyzed using one-way ANOVA.

[0154] 6. Conclusion In the above experiment, the in vivo antitumor effects of sodium porphyrin and targeted photosensitizer X-1 in A549 non-small cell lung cancer subcutaneous transplant tumor model were evaluated. The animal body weight and tumor volume of each experimental group were measured at different time points. Figure 3 and Figure 4 End point tumor weights were as shown Figure 5 shown.

[0155] The above experiments were performed with an average tumor volume of 158 mm 3 The mice were divided into groups at 14:00 and the drugs were administered on the day of grouping. 19 hours after drug administration, the light was irradiated once at a wavelength of 630 nm. The experiment was completed on D22 after grouping. From the above results, it can be seen that at D22, the tumor volume of the negative control group reached 674 mm 3 Compared with the control group, erlotinib at a dose of 25 mg / kg (TV=518 mm 3 , TGI=30.1%, P >0.05) had no significant inhibitory effect on the tumor growth of A549 model. Positive drug sodium porphyrin at a dose of 2 mg / kg (TV=289 mm 3 , TGI=74.7%, P<0.001) had a significant inhibitory effect on the tumor growth of the A549 model. Compared with erlotinib and sodium cytochrome, the mitochondrial targeted photosensitizer X-1 of the present invention (TV=169 mm 3 , TGI=97.8%, P <0.001) showed better efficacy, and the intra-group difference was small, proving that the targeting ability was enhanced and the efficacy was significantly improved.

Claims

1. A mitochondrial targeted photosensitizer or a pharmaceutically acceptable salt thereof, wherein the mitochondrial targeted photosensitizer has the following general formula (I): ， Wherein the groups R each independently represent C 3-9 Heterocyclyl C 1-10 Alkylene-O-, Wherein C 3-9 The heterocyclyl group is unsubstituted or substituted by one or more identical or different groups selected from the group consisting of cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, alkylcarbonyl, haloalkyl or haloalkoxy; The C 1-10 The alkylene group is linear or branched, unsubstituted or substituted by one or more identical or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, alkylcarbonyl, haloalkyl or haloalkoxy, and the alkylene group may be optionally interrupted by one or more heteroatoms selected from S, N and O or by one or more 3-6 membered nitrogen-containing heterocyclic groups; The 3-6 membered nitrogen-containing heterocyclic ring is unsubstituted or substituted by one or more identical or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, haloalkyl or haloalkoxy.

2. The mitochondrial targeting photosensitizer according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the C 3-9 The heterocyclic group contains at least one nitrogen atom selected from the group consisting of pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl, azetidinyl, aziridine, oxazetidinyl, oxaziridine, oxaziridine, oxepanyl, oxazihexaninyl, azepanyl, oxopyrrolidinyl, dioxopyrrolidinyl, oxomorpholinyl, and oxopiperazinyl.

3. The mitochondrial targeted photosensitizer according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the C 1-10 Alkylene is straight chain and unsubstituted.

4. The mitochondrial targeted photosensitizer according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the group R is C 3-6 Heterocyclyl C 1-6 Alkylene-O-, where C 3-6 The heterocyclic group is selected from the group consisting of pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolidinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl, azetidinyl, aziridine, oxazetidinyl, oxaziridine, oxaziridine, oxopyrrolidinyl, dioxopyrrolidinyl, oxomorpholinyl, and oxopiperazinyl; C 1-6 The alkyl group is selected from methylene, ethylene, propylene, n-butylene, n-pentylene or n-hexylene.

5. The mitochondrial targeted photosensitizer according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the group R is C 3-6 Heterocyclyl C 1-4 Alkylene-O-, where C 3-6 The heterocyclic group is selected from the group consisting of pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, oxazine, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolinyl, imidazolidinyl, thiazolidinyl, oxazolidinyl, pyrazolidinyl, azetidinyl, oxazetidinyl, oxaziridinyl, oxaziranyl, oxomorpholinyl, and oxopiperazinyl; C 1-4 The alkylene group is selected from methylene, ethylene, n-propylene or n-butylene.

6. The mitochondrial targeted photosensitizer according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the 3-6 membered nitrogen-containing heterocyclic group is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, furazanyl, dioxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl or morpholinyl.

7. A method for preparing a mitochondrial targeted photosensitizer, comprising the following steps: In the presence of a catalyst and a condensing agent, porphyrinic acid is reacted with a G-OH compound in the presence of a solvent to obtain a product; The GO part in the G-OH compound is defined the same as the R group in the mitochondrial targeting photosensitizer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5.

8. The method according to claim 7, wherein the catalyst is an organic base catalyst selected from N,N-diisopropylethylamine (DIPEA), N,N-dimethylcyclohexylamine, N,N-dimethylbenzylamine, N-methylmorpholine, N-ethylmorpholine, N-methylimidazole, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, pyridine, N,N'-dimethylaminopyridine, 2,6-lutidine or a mixture thereof; The condensing agent is a urea cation type condensing agent, which is selected from O-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TATU), O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate (T BTU), O-(1,2-dihydro-2-oxy-pyridyl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU); carbodiimide type condensing agent: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1,3-dicyclohexylcarbodiimide (DCC); or carbonyl imidazole type condensing agent: N,N-carbonyldiimidazole (CDI); or a mixture thereof, and the molar ratio of the catalyst to the condensing agent is (0.7-3):1; The solvent is selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide, formamide, dimethyl sulfoxide, acetone, pyridine or a mixture thereof.

9. Use of the mitochondrial targeted photosensitizer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6 for preparing a medicament for treating the following diseases: cancer, including reproductive organ cancer, leukemia, mastocytoma, breast cancer, kidney cancer, respiratory tract cancer, brain cancer, digestive tract cancer, urinary tract cancer, liver cancer, eye cancer, skin cancer, head and neck cancer, multiple myeloma, sarcoma or lymphoma; and precancerous lesions of the diseases.

10. A kit comprising the mitochondrial targeting photosensitizer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, which is present in a therapeutically effective amount; and instructions for using the same for photodynamic therapy.

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