Mitochondria-targeted photosensitizer, preparation method therefor and use thereof, and kit comprising same

Abstract

The present application relates to a mitochondria-targeted photosensitizer or a pharmaceutically acceptable salt thereof, wherein the mitochondria-targeted photosensitizer has a structure of general formula (I) as follows, and wherein groups R each independently have the definitions as set forth in the description. The present application further relates to the preparation and use of the mitochondria-targeted photosensitizer, and a kit comprising the photosensitizer.

Description

Mitochondrial-targeting photosensitizers, their preparation methods and uses, and the kits containing them. Technical Field

[0001] This invention belongs to the field of medicinal chemistry, and specifically relates to mitochondrial-targeting photosensitizers, their preparation methods and uses, and a cassette containing them. Background Technology

[0002] Targeted drugs are drugs or their formulations endowed with targeting capabilities. Their purpose is to enable the drug or its carrier to target specific lesion sites and accumulate or release the active ingredient at the target site. Targeted formulations can achieve relatively high drug concentrations at the target site, thereby improving efficacy while suppressing toxic side effects and reducing damage to normal tissues and cells. Based on different targeting mechanisms, drug targeting can be divided into passive targeting, active targeting, and physical targeting. Physical targeting utilizes physical signals such as light, heat, magnetic fields, electric fields, and ultrasound to artificially regulate the distribution and release characteristics of drugs in the body, achieving targeting of lesion sites. Photosensitizers, as targeted drugs that exert their effects through physical targeting, are used in photodynamic therapy (PDT) to effectively treat various diseases. Photodynamic therapy is a novel treatment method. Its principle is a photochemical reaction, and its basic elements are oxygen, a photosensitizer, and visible light. First, the target site selectively absorbs the photosensitizer; then, after local irradiation with light of an appropriate wavelength, the photosensitizer is activated, producing a photosensitizing effect.

[0003] Currently, injectable porphyrin sodium, a novel photosensitizer, is a single active chemical component isolated from photofrin. Preclinical studies in esophageal cancer have shown that injectable porphyrin sodium has the same mechanism of action as photofrin, but achieves the same level of antitumor activity at only 10% of the dosage, with lower phototoxicity. It exhibits significant advantages in efficacy, safety, and quality control. However, previous studies have revealed that porphyrin sodium can selectively accumulate in tumor tissue, but it also distributes throughout the body, accumulating to some extent in normal tissues. Patients need to avoid light for approximately 7 days after treatment. To reduce the phototoxicity of porphyrin sodium, its targeting precision needs to be improved. This study designed a mitochondrial-targeting photosensitizer to enhance the PDT effect under hypoxic conditions, thereby obtaining a highly active, low-toxicity photosensitizer. Summary of the Invention

[0004] In view of the current state of the technology, this invention aims to provide a photosensitizer that can significantly improve tumor targeting, thereby enhancing drug efficacy and reducing toxicity. Its preparation method boasts high yield and a simple synthesis process. The mitochondrial photosensitizer of this invention can be excited under light irradiation of different wavelengths and doses, and exhibits high biosafety, making it a mitochondrial photosensitizer with potential clinical application value and capable of large-scale production.

[0005] On one hand, the present invention provides a mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof, wherein the mitochondrial-targeting photosensitizer has the structure of the following general formula (I):

[0006] Each of the groups R independently represents C. 3-9 Heterocyclic group -O- or C 3-9 Heterocyclic group -C 1-10 alkylene-O-,

[0007] Wherein C 3-9 The heterocyclic group is unsubstituted or substituted by one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, alkylcarbonyl, dialkylamino, haloalkyl or haloalkoxy;

[0008] The C 1-10 The alkylene group is straight-chain or branched, unsubstituted, or substituted with one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, alkylcarbonyl, dialkylamino, haloalkyl, or haloalkoxy, and the alkylene group may optionally be separated by one or more heteroatoms selected from S, N, and O or by one or more 3-6 membered nitrogen-containing heterocyclic groups.

[0009] The 3-6 membered nitrogen-containing heterocycles therein are unsubstituted or substituted by one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, haloalkyl or haloalkoxy.

[0010] On the other hand, the present invention provides a method for preparing a mitochondrial-targeting photosensitizer of formula (I), which includes the following steps: reacting porphyrinic acid with a G-OH compound in the presence of a solvent in the presence of a catalyst and a condensing agent to obtain the product;

[0011] The GO moiety in the G-OH compound is defined in the same way as the R group described above.

[0012] On the other hand, the present invention provides the use of mitochondrial-targeting photosensitizers or pharmaceutically acceptable salts thereof for the preparation of medicaments for treating the following diseases: cancers, including cancers of the reproductive organs (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer, and testicular cancer), leukemia, mast cell tumor, breast cancer, kidney cancer, respiratory tract cancers (such as lung cancer, particularly small cell and non-small cell lung cancer, as well as bronchial cancer and pleural pulmonary blastoma), brain cancer, digestive tract cancers (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small bowel cancer, and salivary gland cancer), urinary tract cancer, liver cancer, eye cancers (e.g., intraocular melanoma and retinoblastoma), skin cancer, head and neck cancers (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, and oral cancer, as well as squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and precancerous lesions of the above diseases.

[0013] In another aspect, the present invention provides a method for treating diseases comprising administering to a subject in need a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof; wherein the diseases are selected from cancers, including cancers of the reproductive organs (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer, and testicular cancer), leukemia, mast cell tumor, breast cancer, kidney cancer, respiratory tract cancers (such as lung cancer, particularly small cell and non-small cell lung cancer, and bronchial ... Cancers including pleural and pulmonary blastoma, brain cancer, gastrointestinal cancers (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small bowel cancer, and salivary gland cancer), urinary tract cancer, liver cancer, eye cancers (such as intraocular melanoma and retinoblastoma), skin cancer, head and neck cancers (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, oral cancer, and squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and precancerous lesions of the above conditions.

[0014] In another aspect, the present invention provides a kit containing a therapeutically effective amount of the mitochondrial-targeting photosensitizer of formula (I) of the present invention or a pharmaceutically acceptable salt thereof; and instructions for use therein in photodynamic therapy.

[0015] Surprisingly, the inventors discovered 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. Its preparation method has high yield and simple synthesis process, and it can be used as a highly efficient photosensitizer with potential clinical application value and can be produced on a large scale. Attached Figure Description

[0016] Figure 1 shows a scatter plot of fluorescence imaging and colocalization analysis of DVDMS and MTDR.

[0017] Figure 2 shows a scatter plot of fluorescence imaging and colocalization analysis of compound X-1 with MTDR.

[0018] Figure 3 shows the weight changes of A549 tumor-bearing mice after drug administration.

[0019] Figure 4 shows the changes in tumor volume in A549 tumor-bearing mice after drug administration.

[0020] Figure 5 shows the tumor weight at each experimental endpoint in A549 tumor-bearing mice. Detailed Implementation

[0021] The invention will be described in more detail below.

[0022] Unless otherwise stated, “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 (hereinafter referred to as photosensitizer)” are used synonymously.

[0023] 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.

[0024] Unless otherwise defined, the name of a chemical group should generally be understood as such that its connection to the skeleton or the rest of the molecule is through the structural elements of the last mentioned related chemical group, i.e.,, for example, through the carbon atoms of the alkylene group in the case of heterocyclic alkylene groups.

[0025] As used herein, the terms “optional,” “optional,” or “optionally” mean that the event, situation, or substance described below may or may not occur or not exist, and such description includes both the occurrence and absence of the event, situation, or substance.

[0026] Unless otherwise stated, the following definitions apply to groups or substituents used throughout this specification and claims. Within the scope of this invention, the meanings of all repeated groups are independent of each other.

[0027] As used in this article, the term "cyano" refers to a functional group formed by the connection of carbon and nitrogen atoms through a triple bond.

[0028] As used herein, the term "halogen" refers to, for example, fluorine, chlorine, bromine, or iodine. If the term is used with a group, "halogen" refers to, for example, a fluorine, chlorine, bromine, or iodine atom.

[0029] As used herein, the term "alkyl" refers in each case to a saturated straight-chain or branched hydrocarbon group having a specified number of carbon atoms, for example (C1-C2). 10(C1-C6)-alkyl and (C1-C4)-alkyl, examples including but 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.

[0030] 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 identical or different halogen atoms, for example (C1-C2). 10 (C1-C6)-haloalkyl, (C1-C4)-haloalkyl, and 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-trifluoropropyl-2-yl. Fluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, and 1,1,1-trifluoropropyl-2-yl are preferred.

[0031] According to the present invention, "cycloalkyl"—either on its own or as part of a chemical group—preferably represents a monocyclic, bicyclic, or tricyclic hydrocarbon having 3 to 10 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclic [2.2.1]heptyl, bicyclic [2.2.2]octyl, or adamantyl. Cycloalkyl groups having 3, 4, 5, 6, or 7 carbon atoms are also preferred, for example, especially cyclopropyl or cyclobutyl. The cycloalkyl groups of the present invention can be substituted with one or more identical or different groups.

[0032] According to the present invention, "alkoxy" preferably represents a straight-chain or branched alkyl-O- having 1 to 6 carbon atoms, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy. Alkoxy having 1 to 4 carbon atoms is also preferred. The alkoxy groups of the present invention can be substituted with one or more identical or different groups.

[0033] As used herein, the term "haloalkoxy" refers to an alkoxy group in which one or more hydrogen atoms of the alkyl portion are replaced by one or more identical or different halogen atoms, for example (C1-C2). 10 (C1-C6)-haloalkoxy, (C1-C4)-haloalkoxy, and 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, and pentafluoroethoxy. Fluoromethoxy, difluoromethoxy, trifluoromethoxy, 1-fluoroethoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, and pentafluoroethoxy are preferred.

[0034] According to the present invention, "alkathioyl" preferably represents a straight-chain or branched alkyl-S- having 1 to 6 carbon atoms, such as methylthioyl, ethylthioyl, n-propylthioyl, isopropylthioyl, n-butylthioyl, isobutylthioyl, sec-butylthioyl, and tert-butylthioyl. Alkathioyl having 1 to 4 carbon atoms is also preferred. The alkathioyl group of the present invention can be substituted with one or more identical or different groups.

[0035] According to the present invention, "alkyl carbonyl" preferably represents a straight-chain or branched alkyl-C(=O)- having 2 to 7 carbon atoms, such as methyl carbonyl, ethyl carbonyl, n-propyl carbonyl, isopropyl carbonyl, sec-butyl carbonyl, and tert-butyl carbonyl. Alkyl carbonyl having 1 to 4 carbon atoms is also preferred. The alkyl carbonyl of the present invention can be substituted with one or more identical or different groups.

[0036] According to the present invention, a "heterocyclic group" represents a carbocyclic system having at least one ring in which at least one carbon atom is substituted by a heteroatom, preferably selected from N, O, S, P, B, Si, and Se, and is saturated, unsaturated, or heteroaromatic, and can be unsubstituted or substituted, wherein the bonding site is on a ring atom. Unless otherwise defined, a heterocycle comprises 3 to 9 ring atoms (i.e., C atoms). 3-9 Heterocyclic groups), especially those with 3 to 6 ring atoms (i.e., C164-C ... 3-6The heterocycle contains one or more, preferably 1 to 4, especially 1, 2 or 3 heteroatoms, preferably selected from N, O and S, but the two oxygen atoms should not be directly adjacent. The heterocycle typically contains no more than 4 nitrogen atoms and / or no more than 2 oxygen atoms and / or no more than 2 sulfur atoms. In the case of optionally substituted heterocycles, the invention also includes polycyclic systems, such as 8-azabicyclo[3.2.1]octyl, 1-azabicyclo[2.2.1]heptyl, 1-oxa-5-azaspiro[2.3]hexyl or 2,3-dihydro-1H-indole.

[0037] The heterocyclic groups of the present invention are, for example: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazolyl, dioxazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetraazinyl, oxazinyl, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, dioxacyclohexyl, pyrrolinyl, pyrrolylalkyl, imidazoline. The compounds include: alkyl, imidazoalkyl, thiazoalkyl, oxazolidinyl, dioxopentyl, dioxacyclopentenyl, pyrazolyl, tetrahydrofuranyl, dihydrofuranyl, oxacyclobutyl, oxacyclopropyl, aziridine, aziridinepropyl, oxacyclobutyl, oxacyclopropyl, oxacyclobutyl, oxacyclopropyl, oxacycloheptyl, oxacyclohexyl, aziridineheptyl, oxopyridine, dioxopyridine, oxomorpholinyl, oxopyridine, and oxacycloheptyl.

[0038] According to the present invention, “C” 3-9 "heterocyclic group" and "C" 3-9 Heterocyclic C 1-10 The "heterocyclic group" in "alkylene" has the meanings described above; "C" 1-10 The term "alkylene" refers to methylene, ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, nonylene, and decylene, and their isomers.

[0039] According to the present invention, the "3-6 member nitrogen-containing heterocyclic group" is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridinyl, pyridinyl, pyrazinyl, piperidinyl, triazinyl, tetraazinyl, 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 furazonyl.

[0040] The compounds of formula (I) mentioned anywhere in this document also cover any diastereomers or enantiomers and E / Z isomers of the presence of compounds of formula (I), as well as their salts.

[0041] Depending on the nature of the substituents, compounds of formula (I) described anywhere herein may also be in the form of stereoisomers, i.e., geometric and / or optical isomers or mixtures of isomers with different compositions. This invention provides pure stereoisomers and any desired mixtures of these isomers, although only compounds of formula (I) are generally discussed herein.

[0042] If appropriate, the compound of formula (I) can exist in various polymorphic forms or as mixtures of various polymorphic forms. Pure polymorphs and mixtures of polymorphs are provided by the present invention and can be used according to the present invention.

[0043] This document does not cover compounds obtained from combinations that contradict the laws of nature and that would therefore be excluded by a person skilled in the art based on his / her expertise. For example, ring structures with three or more adjacent oxygen atoms are excluded.

[0044] According to one aspect of the present invention, a mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof is provided, wherein the mitochondrial-targeting photosensitizer has the structure of the following general formula (I):

[0045] Each of the groups R independently represents C. 3-9 Heterocyclic group -O- or C 3-9 Heterocyclic C 1-10 alkylene-O-,

[0046] Wherein C 3-9 The heterocyclic group is unsubstituted or substituted by one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, alkylcarbonyl, dialkylamino, haloalkyl or haloalkoxy;

[0047] The C 1-10 The alkylene group is straight-chain or branched, unsubstituted, or substituted with one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, alkylcarbonyl, dialkylamino, haloalkyl, or haloalkoxy, and the alkylene group may optionally be separated by one or more heteroatoms selected from S, N, and O or by one or more 3-6 membered nitrogen-containing heterocyclic groups.

[0048] The 3-6 membered nitrogen-containing heterocycles therein are unsubstituted or substituted by one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, haloalkyl or haloalkoxy.

[0049] Pharmaceutically acceptable salts of compounds of formula (I) of this invention refer to those salts that are pharmaceutically considered safe and suitable for use in pharmaceutical formulations. These salts are typically generated by reacting compounds of formula (I) with acids or bases to improve the solubility, stability, and bioavailability of the drug.

[0050] Pharmaceutically acceptable salts of the compounds of this invention are, for example, products obtained by reacting compounds of formula (I) with acids. Preferably, the acids include, but are 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 pharmaceutically acceptable basic addition salts of the compounds of formula (I) of this invention include 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.

[0051] In a preferred embodiment of the invention, each group R in formula (I) has the same definition.

[0052] In a preferred embodiment of the invention, wherein in formula (I), C 3-9 The heterocyclic group is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, piperidinyl, triazinyl, tetraazinyl, oxazinyl, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, dioxacyclohexyl, pyrrolinyl, pyrrolylalkyl, imidazolinyl Imidazolidinyl, thiazolyl, oxazolidinyl, dioxopentenyl, dioxacyclopentenyl, pyrazolyl, tetrahydrofuranyl, dihydrofuranyl, oxacyclobutyl, oxacyclopropane, aziridine, aziridinepropane, oxacyclobutyl, oxacyclopropane, oxacyclobutyl, oxacyclopropane, oxacycloheptyl, oxacyclohexyl, aziridineheptyl, oxopyridine, dioxopyridine, oxomorpholinyl, oxopyridine, and oxacycloheptyl.

[0053] In a preferred embodiment of the invention, wherein in formula (I), C 1-10 The alkyl group is straight-chain and unsubstituted.

[0054] In a preferred embodiment of the invention, wherein in formula (I), C 3-9The heterocyclic group contains at least one nitrogen atom and is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazolyl, dioxazolyl, isothiazolyl, pyridinyl, pyridinyl, pyrazinyl, triazinyl, tetraazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolylalkyl, imidazolinyl, imidazolyl, thiazolyl, oxazolylalkyl, pyrazolylalkyl, azacyclic butyl, azacyclic propyl, oxazcyclic butyl, oxazcyclic propyl, oxazcyclic heptyl, oxazcyclic hexyl, azacyclic heptyl, oxopyrrolyl, dioxopyrrolyl, oxomorpholinyl, and oxopiperazinyl.

[0055] In a preferred embodiment of the invention, wherein in formula (I), group R is C 3-6 Heterocyclic 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, oxtriazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridinyl, pyridinyl, pyrazinyl, triazinyl, tetrazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrololinyl, pyrrolylalkyl, imidazolinyl, imidazolyl, thiazolyl, oxazolylalkyl, pyrazolylalkyl, azahexacyclobutyl, azahexacyclopropyl, oxazhexacyclobutyl, oxazhexacyclopropyl, oxazhexacyclohexyl, oxopyrrolylalkyl, dioxopyrrolylalkyl, oxomorpholinyl, and oxopiperazinyl, preferably piperazinyl, morpholinyl, thiomorpholinyl, pyrrololinyl, pyrrolylalkyl, imidazolinyl, imidazolyl, thiazolyl, oxazolylalkyl, and pyrazolylalkyl; C 1-6 The alkylene group is selected from methylene, ethylene, propylene, n-butylene, n-pentylene, or n-hexylene.

[0056] In a preferred embodiment of the invention, wherein in formula (I), group R is C 3-6 Heterocyclic C 1-4 alkylene-O-, where C 3-6 The heterocyclic group is selected from the following groups: pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridazinyl, pyrazinyl, oxazinyl, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolinyl, imidazoalkyl, thiazoalkyl, oxazolalkyl, pyrazolalkyl, azirheptanyl, oxazirheptanyl, oxazirheptanyl, oxomorpholinyl, and oxopiperazinyl, preferably morpholinyl, oxazinyl, isoxazinyl, oxazirheptanyl, oxazirheptanyl, and oxazirheptanyl; C 1-4The alkylene group is selected from methylene, ethylene, propylene, or n-butylene. In a preferred embodiment of the invention, in formula (I), the group R is, for example, morpholino-N-ethyl.

[0057] In a preferred embodiment of the invention, the 3-6 member nitrogen-containing heterocyclic group is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridinyl, pyridinyl, pyrazinyl, piperidinyl, triazinyl, tetraazinyl, 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 furazonyl.

[0058] In a preferred embodiment of the invention, wherein in formula (I), the mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof is used to treat the following diseases: cancers, including cancers of the reproductive organs (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer, and testicular cancer), leukemia, mast cell tumor, breast cancer, kidney cancer, respiratory cancers (such as lung cancer, particularly small cell and non-small cell lung cancer, as well as bronchial cancer and pleural pulmonary blastoma). Cancers include: brain cancer, digestive tract cancers (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small bowel cancer, and salivary gland cancer), urinary tract cancer, liver cancer, eye cancer (such as intraocular melanoma and retinoblastoma), skin cancer, head and neck cancers (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, oral cancer, and squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and precancerous lesions of the above conditions.

[0059] In this article, "precancerous lesions" refer to abnormal or excessive cell proliferation, which has a high probability of becoming cancerous.

[0060] The definitions of the groups listed above in general terms or within the preferred range can be combined with each other as needed, i.e., combinations between the given preferred ranges.

[0061] The following compounds of formula (I) of the present invention are particularly preferred.

[0062] The R group is morpholinoethylidene-O-.

[0063] According to another aspect of the invention, the invention also provides a method for preparing a compound of formula (I), comprising the following steps:

[0064] In the presence of a catalyst and a condensing agent, porphyrinic acid was reacted with a G-OH compound in the presence of a solvent to obtain the product;

[0065] The GO moiety in the G-OH compound is defined in the same way as the R group described above.

[0066] In the method of the present invention, the catalyst used is an organic base catalyst, which is selected from N,N-diisopropylethylamine (DIPEA), N,N-dimethylcyclohexylamine, N,N-dimethylbenzylamine, N-methylmorpholine, N-ethylmorpholine, N-methylimidazolium (NMI), 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, pyridine, N,N-dimethylaminopyridine, 2,6-dimethylpyridine or mixtures thereof, more preferably N-methylimidazolium; the organic solvent used is selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide, formamide, dimethyl sulfoxide, acetone, pyridine or mixtures thereof, more preferably N,N-dimethylformamide.

[0067] In the method of this invention, the condensing agent used is a urea cation type condensing agent, selected from O-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), O-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TATU), and O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluorophosphate. Boronate esters (TBTU), O-(1,2-dihydro-2-oxopyridyl)-1,1,3,3-tetramethylurea tetrafluoroborate (TPTU); carbodiimide-type condensing agents, such as 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1,3-dicyclohexylcarbodiimide (DCC); or carbonylimidazolium-type condensing agents, such as N,N-carbonyldiimidazole (CDI); or mixtures thereof, preferably selected from HATU, EDCI, CDI, or mixtures thereof.

[0068] In the method of the present invention, the molar ratio of the catalyst to the condensing agent is (0.7-2.0):1, preferably (0.8-1.8):1, and more preferably (0.9-1.5):1.

[0069] The method for preparing compounds of formula (I) of the present invention uses a specific combination of an organic base catalyst and a specific condensing agent, particularly a specific ratio of N-methylimidazole and HATU, which can obtain the target compound with high purity in good yield through simple post-processing. Too little or too much catalyst or condensing agent 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.

[0070] In the method of the present invention, the molar ratio of the 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).

[0071] The molar ratio of the porphyrin to the catalyst is 1:(4.0-12.0), preferably 1:(6.0-10.0), and more preferably 1:(8.0-9.0).

[0072] In the method of the present invention, the condensation reaction time is 0.5-24 hours, preferably 1-20 hours, and more preferably 1-18 hours.

[0073] Another aspect of the present invention provides the use of the aforementioned mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof in the preparation of medicaments for treating the following diseases: cancers, including cancers of the reproductive organs (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer, and testicular cancer), leukemia, mast cell tumor, breast cancer, kidney cancer, respiratory tract cancers (such as lung cancer, particularly small cell and non-small cell lung cancer, as well as bronchial cancer and pleural pulmonary blastoma), brain cancer, digestive tract cancers (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small bowel cancer, and salivary gland cancer), urinary tract cancers, liver cancer, eye cancers (e.g., intraocular melanoma and retinoblastoma), skin cancer, head and neck cancers (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, and oral cancer, as well as squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and hyperplastic conditions of the above diseases, such as precancerous lesions.

[0074] Another aspect of the present invention provides a method for treating the following diseases, the method comprising administering to a subject in need a therapeutically effective amount of a compound of formula (I) above or a pharmaceutically acceptable salt thereof; wherein the disease being treated is selected from: cancers, including cancers of the reproductive organs (such as cervical cancer, ovarian cancer, endometrial cancer, vaginal cancer, vulvar cancer, uterine sarcoma, prostate cancer, and testicular cancer), leukemia, mast cell tumor, breast cancer, kidney cancer, respiratory tract cancers (such as lung cancer, particularly small cell and non-small cell lung cancer, as well as bronchial cancer and...). Pleural blastoma, brain cancer, gastrointestinal cancers (such as esophageal cancer, anal cancer, colon cancer, colorectal cancer, gallbladder cancer, stomach cancer, pancreatic cancer, rectal cancer, small bowel cancer, and salivary gland cancer), urinary tract cancer, liver cancer, eye cancer (such as intraocular melanoma and retinoblastoma), skin cancer, head and neck cancers (such as thyroid cancer, parathyroid cancer, laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer, lip cancer, oral cancer, and squamous cell carcinoma), multiple myeloma, sarcoma, or lymphoma; and hyperplastic conditions of the above diseases, such as precancerous lesions.

[0075] As used herein, the term “therapeutic effective amount” refers to the amount of an active compound or agent that elicits a biological or pharmaceutical response sought or desired by researchers, physicians or other clinicians in a tissue, system, animal, individual or human.

[0076] As used herein, the term “treatment” means killing, inhibiting, or slowing the growth or increase in size of a cluster or group of overproliferating cells or tumor or cancerous growth, reducing the number of overproliferating cells, or preventing its spread to other anatomical sites, and reducing the size of overproliferative growth or the number of overproliferating cells. However, it should be understood that “treatment” does not necessarily mean a cure or complete elimination of overproliferative growth.

[0077] Another aspect of the present invention provides a kit containing a therapeutically effective amount of the mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof; and instructions for use therein in photodynamic therapy.

[0078] As used herein, the term "pharmacy box" means any commercial packaging containing a container for holding the compounds of the present invention or pharmaceutical preparations containing the present invention, and optionally also includes separate containers such as separate vials or separate foil packs, for example, for holding reconstitution dissolution matrices. The containers may be any conventional shape or form known in the art and are made of pharmaceutically acceptable materials.

[0079] In a preferred embodiment, the kit provided by the present invention comprises a lyophilized formulation of the compound of the present invention in a therapeutically effective amount for treating the disease as described above; a dissolving matrix (such as water for injection containing excipients such as cosolvents) for reconstituted the lyophilized formulation for application; and instructions for using the compound of the present invention as a photosensitizer in photodynamic therapy.

[0080] The various components of the kit, such as the compounds of the present invention or their pharmaceutically acceptable salts, pharmaceutical preparations containing them, dissolving matrices, and other active ingredients for treating cancer or precancerous lesions, may be packaged in individual containers. Regardless of the number or type of containers, the kit may also include a device for assisting in the administration of the medication to the patient. This device may be a patch, inhaler, syringe, suction tube, spoon with measuring units, or any approved medical delivery device.

[0081] The photosensitizer of the present invention can be administered in any of the following ways: orally, via oral mucosa, via spray inhalation, via rectal administration, via nasal administration, via vaginal administration, via local administration, via enteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, or intracranial injection or infusion, or via an external reservoir, wherein local, intraperitoneal, or intravenous administration is preferred.

[0082] The photosensitizer of this invention can be administered in unit dose form. The dosage form can be liquid, semi-solid, or solid. Liquid dosage forms can be true solutions, colloids, microparticles, or suspensions. Semi-solid dosage forms can be ointments, creams, pastes, gels, etc. Other dosage forms include, for example, tablets, capsules, pellets, aerosols, pills, powders, solutions, emulsions, granules, suppositories, lyophilized powder injections, inclusion complexes, implants, patches, liniments, and sustained-release formulations.

[0083] For oral preparations, tablets and capsules can be prepared using pharmaceutically known methods, and tablets can also be coated; oral liquids can be prepared as suspensions, solutions, emulsions, syrups, or as dry products, to be replenished with water or other suitable medium before use. Flavoring or coloring agents can be added if necessary.

[0084] For parenteral administration, the liquid dosage form is typically made from the drug of this invention and a sterilized carrier. Water is the preferred carrier. Depending on the chosen carrier and drug concentration, the drug can be either soluble in the carrier or prepared as a suspension. When preparing an injectable solution, the drug is first dissolved in water, filtered, sterilized, and then packaged into a sealed bottle or ampoule.

[0085] It can also be formulated into sterile injectable preparations, including crystalline powder injections and lyophilized powder injections.

[0086] When applied topically to the skin or mucous membranes, the medicine of the present invention can be formulated into suitable ointments, lotions, gels, or pastes, wherein the active ingredient is suspended or dissolved in one or more carriers. Alternatively, it can be formulated into forms for use with devices such as microneedles.

[0087] The photosensitizer of the present invention also comprises a pharmaceutically acceptable carrier, excipients, and / or other adjuvants. When comprising a pharmaceutically acceptable carrier, excipients, and / or other adjuvants, an effective dose of the photosensitizer of the present invention or a pharmaceutically acceptable salt thereof, along with one or more pharmaceutically acceptable carriers, excipients, and / or other adjuvants, is typically combined to form a suitable administration or dosage form. This procedure includes mixing, granulating, compressing, dissolving, or lyophilizing the components by suitable methods. The carrier content in the drug can be from 1 to 98% by weight. For convenience, other adjuvants such as local anesthetics, preservatives, and buffers may be directly dissolved in the carrier.

[0088] The photosensitizer of the present invention can be prepared into injections, topical formulations, or oral formulations using methods known to those skilled in the art. The photosensitizer of the present invention is preferably an injection, administered intravenously.

[0089] The pharmaceutically acceptable carriers, excipients, and / or other adjuvants that can be used to prepare the photosensitizers of the present invention or their application forms are conventional pharmaceutically acceptable carriers, excipients, and / or adjuvants known to those skilled in the art for this purpose.

[0090] 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 being treated, the wavelength, power and duration of light used for treatment, and the type and severity of the disease being treated. This optimal dosage can be determined using conventional techniques. The optimal course of treatment, i.e., the daily dose of the compound or drug of the present invention within a specified time period, can be determined using methods known in the art.

[0091] The photosensitizer of the present invention is used at a dosage of 0.01-100.0 mg / kg of subject, for example 0.01-50.0 mg / kg of subject, preferably 0.05-10.0 mg / kg of subject, particularly 0.1-6.0 mg / kg of subject, 0.2-5.0 mg / kg of subject; and is irradiated with laser light at a wavelength of 300-800 nm, preferably 600 nm-650 nm, particularly 630 nm, usually once; the light dose is 1-300 J, preferably 20-200 J, more preferably 50-150 J; the light power of the laser as the light source is usually 50-800 mW, preferably 100-500 mW; the irradiation time is 10-2400 seconds, preferably 60-1800 seconds, more preferably 100-1500 seconds; the irradiation delay is usually 4-48 hours, preferably 10-25 hours.

[0092] In the context of this invention, the term "optical power" refers to the actual optical power of the light-irradiated area, which is measured by an optical power meter to determine the actual power of the treatment area.

[0093] In the context of this invention, the term "light dose" refers to the actual light dose at the irradiated site, obtained by multiplying the light power by the irradiation time.

[0094] In this invention, unless otherwise expressly stated, all contents and percentages in the context of this application are based on weight; unless otherwise expressly stated, the method steps of this invention are performed at room temperature and pressure. Unless otherwise expressly stated, the reagents used in the following synthesis examples are all commercially available conventional products.

[0095] In the method for preparing the compound of formula (I) of this invention, the reaction progress is monitored using normal-phase silica gel thin-layer chromatography (TLC), and the completion of the reaction is determined by observing the gradual disappearance of the starting material spots. This detection is performed on a thin-layer chromatograph. The thin-layer chromatograph that can be used in the method of this invention is a commercially available conventional thin-layer chromatograph, such as the three-in-one ultraviolet analyzer, model: WHF-204B, purchased from Shanghai Heqi Glass Instrument Co., Ltd.

[0096] In the above-described method for preparing the compounds of the present invention, the method may optionally include other post-processing steps. These post-processing steps may include, for example, conventional purification steps such as pH adjustment, crystallization, extraction, filtration, vacuum concentration, and drying. All of the above steps can be performed in a conventional manner known to those skilled in the art. If present, extraction is typically performed using a mixed solution of dichloromethane and methanol, preferably a dichloromethane / methanol mixture of 10:1 (v / v); drying is typically performed by freeze-drying, infrared drying, vacuum drying, etc., preferably freeze-drying.

[0097] In a preferred embodiment of the present invention, the method may further include a purification step via a chromatographic column. The purification may be performed using a normal-phase silica column with silica particle size of 30-100 μm, preferably 40-63 μm, and a loading of 20-120 g, preferably 40 g, eluted with dichloromethane / methanol (v / v) (elution gradient 100% / 0% to 90% / 10%, gradient elution time 15 min).

[0098] Unless otherwise expressly stated, all operations are performed at room temperature; the reagents used are commercially available or prepared by methods known to those skilled in the art.

[0099] The compounds of the present invention can be prepared according to the methods described above. However, it should be understood that those skilled in the art, based on their common sense and available publications, can adjust the methods according to the specific circumstances of the compounds of the present invention to be synthesized.

[0100] Detailed synthetic examples of the selected compounds of the present invention are given below. However, these examples are merely illustrative and should not be construed as limiting the scope of the invention in any way.

[0101] The peaks in the synthesis examples are recorded in the form of an NMR peak list. 1 H NMR spectral data were obtained on a Bruker 400 MHz NMR spectrometer, and the listed signals have the following meanings: s = singlet, d = doublet, dd = doublet, m = multiplet, br s = broad singlet. The deuterated solvent used in each case is also... 1 Specify in the H NMR spectral data.

[0102] In this invention, in addition to the above-mentioned NMR peak list, 1 In addition to 1H NMR spectral data, the structures of the compounds prepared in the synthesis examples were characterized by liquid chromatography-mass spectrometry (LC-MS). LC-MS data were obtained using a Waters instrument (model: SQD2).

[0103] Synthesis Examples

[0104] Synthetic reagents and equipment

[0105] Table 1. Synthetic Reagents

[0106] Table 2: Synthesis Equipment

[0107] Synthesis of compound X-1

[0108] At room temperature (20°C), porphyrin (50 mg, 0.044 mmol, 1.0 equ) was dissolved in 5 mL of DMF. Then, HATU (133 mg, 0.350 mmol, 8.0 equ) and N-methylimidazole (29 mg, 0.350 mmol, 8.0 equ) were added sequentially at 20°C. After reacting for 1 hour at room temperature, N-(2-hydroxyethyl)morpholine (34 mg, 0.263 mmol, 6.0 equ) was added, and the reaction was carried out at room temperature for 12 hours. Normal phase thin-layer chromatography was used to detect the reaction until the starting material was completely reacted. The reaction solution was poured into 100 mL of water and extracted three times (50 mL × 3) with a mixed solution of dichloromethane / methanol (10 / 1, V / V). The organic phases were combined and then washed with 200 mL of saturated sodium chloride aqueous solution. After drying with anhydrous sodium sulfate, the solution was filtered and concentrated. The solution was then purified by passing it through 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%).

[0109] LC-MS (m / z): 1597.1 [M+H] +

[0110] 1 H NMR (400MHz, DMSO-d6): δ10.36(s,2H),10.25(s,2H),9.90(s,2H),9.29(br s,2H),8.48(dd,J=18.0,11.6Hz,2H),6.44(d,J=18.0Hz,2H)),6.22-6.18(m,4H),4.4 6-4.40(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).

[0111] Target capability evaluation

[0112] Using human lung cancer cell line A549 as the research object, the subcellular localization (mitochondrial targeting) and distribution of mitochondrial porphyrin sodium (DVDMS) and the targeted photosensitizer X-1 in tumor cells were detected.

[0113] 1. Materials and Equipment

[0114] Table 3 Material List

[0115] Table 4 Equipment List

[0116] 2. Experimental System

[0117] Cell information: A549 cells (catalog number: FH0045) were purchased from Fuheng Biotechnology.

[0118] Cell culture: Cells were removed from the -80℃ freezer and revived, then cultured at 37℃ until they reached a good growth state. Cells were grown in RPMI-1640 complete medium containing 10% fetal bovine serum, 1% penicillin (10,000 U / mL) and streptomycin (10,000 μg / mL) in a CO2 incubator at 37℃, saturated humidity, and 5% CO2.

[0119] 3. Experimental Methods

[0120] 3.1 Drug Preparation

[0121] Table 5 Drug Preparation Table

[0122] 3.2 Cell drug administration and staining

[0123] 1. Prepare DVDMS and X-1 working solutions (4 μg / mL) using RPMI-1640 medium, respectively.

[0124] 2. After cell digestion and resuspending, the cells were diluted to 1.5 × 10⁻⁶ using RPMI-1640 complete medium. 5 Cells / mL. Add 1 mL of cell suspension to a glass-bottomed culture dish and incubate at 37°C for at least 20 hours to allow the cells to adhere.

[0125] 3. 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 petri dishes respectively, and incubate at 37°C in the dark for 4 hours.

[0126] 4. After 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.

[0127] 5. Prepare MTDR working solution.

[0128] 6. Remove the old culture medium, add the diluted 250 nM MTDR working solution, and incubate at 37°C for 15 minutes for staining.

[0129] 7. Remove the old culture medium and add 1 mL of serum-free and phenol red-free RPMI-1640 culture medium. Repeat the operation twice.

[0130] 8. Remove the old culture medium, add 1 mL of general-purpose tissue fixative (neutral), and fix the cells at room temperature for 10 minutes.

[0131] 9. Remove the fixative, add 1 mL of PBS, and repeat the operation twice.

[0132] 3.3 Imaging cells using fluorescence microscopy

[0133] 1. Locate the cells using bright field, adjust the objective lens to 40x, and adjust the focus to make the mitochondria clear.

[0134] 2. Excite the fluorescence channel with a 640nm light source and receive it with a 690nm light source, observe the fluorescence of the MTDR, and take an image.

[0135] 3. Excite the fluorescence channel with a 405nm light source and receive it with a 630nm fluorescence channel, observe the fluorescence of DVDMS and X-1, and take images.

[0136] 3.4 Experimental Results

[0137] When DVDMS or X-1 is excited by a 405nm light source, good fluorescence imaging can be observed in the 630nm receiving channel. The imaging results and colocalization analysis scatter plots are shown in Figure 1 and Figure 2, respectively.

[0138] As shown in Figure 1, the mitochondrial fluorescent probe MTDR clearly labeled the structure of mitochondria within the cell. DVDMS was mainly enriched around the cell nucleus, where mitochondria were also concentrated. However, the colocalization of DVDMS and MTDR was weak, with a Pearson correlation coefficient of 0.376, indicating poor mitochondrial targeting.

[0139] As shown in Figure 2, X-1 and MTDR have good co-localization, and both can clearly show the structure of mitochondria. The Pearson correlation coefficient is 0.901, which can be considered that X-1 has good mitochondrial targeting.

[0140] As shown in Figures 1 and 2, compared with DVDMS, X-1 has better mitochondrial targeting ability, and is expected to improve its ability to target and photodynamically kill tumors in vivo, enhance the therapeutic effect and reduce the toxic and side effects.

[0141] 4. In vivo efficacy evaluation

[0142] 4.1 Experimental design

[0143] Table 6 Experimental design

[0144] Note: Administration volume: 10 μL / g according to the body weight of mice

[0145] 4.2 Materials and equipment

[0146] Table 7 Material list

[0147] Table 8 Equipment list

[0148] 4.3 Experimental animals and feeding management

[0149] Experimental animals:

[0150] Strain: Balb / c-nude mice;

[0151] Week age: 6 - 8 weeks old; <000​​​​​​​​​​​​​​​​​​​​​​​​​​​√Humidity: 30-70%

[0162] √ Photoperiod: 12 hours of light, 12 hours of darkness

[0163] Cage: Made of polycarbonate. Bedding is corn cob, changed weekly.

[0164] Food: Laboratory animals were allowed free access to food throughout the experimental period (sterilized by irradiation, dry granular food).

[0165] Drinking water: Laboratory animals may drink sterilized water freely.

[0166] Cage labeling: Each cage animal information card should indicate the number of animals in the cage, sex, strain, date of receipt, administration regimen, experiment number, group, and experiment start date.

[0167] Animal identification: Ear clipping method.

[0168] 4.4 Experimental Methods

[0169] 1. Cell Culture

[0170] A549 cells were cultured in vitro in adherent medium with 10% fetal bovine serum at 37°C in a 5% CO2 incubator. Routine passages were performed twice a week. Trypsin-EDTA (0.25%) solution was used. When cell saturation reached 80%-90% and the desired number was achieved, cells were harvested, counted, and the cell density was adjusted to 5 × 10⁶ cells / day using PBS. 7 cells / ml for later use.

[0171] 2. Animal inoculation

[0172] Animals were placed in the animal facility for a 7-day acclimatization period before experiments could be conducted. 0.1 mL (5.0 × 10⁻⁶) of [the solution was used]. 6 A549 cells were subcutaneously inoculated into the right posterior back of each mouse, near the thigh.

[0173] 3. Routine observation of laboratory animals

[0174] The use and welfare of laboratory animals are conducted in accordance with the guidelines of the International Committee for Assessment and Accreditation of Laboratory Animals (AAALAC). Animal health and mortality are monitored daily, and routine checks include observing the effects of tumor growth and drug treatment on daily behavior such as activity levels, food and water intake (visual assessment only), physical appearance, or other abnormalities.

[0175] 4. Compound preparation

[0176] Table 9. Compound Formulation Table

[0177] 4.5. Random Grouping

[0178] After cell seeding, tumor growth was monitored regularly. Tumors were observed after 21 days of growth, reaching an average tumor volume of 158 mm. 3 At the time of the experiment, 24 animals with suitable tumor size were randomly selected and randomly divided into 4 groups of 6 animals each, according to tumor size. Drug administration began on the day of grouping, designated D0. Except for the erlotinib group, which received daily gavage, the other groups received a single intravenous administration via tail vein. Specific administration protocols are shown in Table 6. All drug solutions were prepared fresh before the experiment and refrigerated in the dark before use. Except for the tumor illumination stage, tumor-bearing animals were strictly protected from light for 3 days after intravenous administration, followed by low-light housing.

[0179] 4.6. Light

[0180] Groups G3-G4 received phototherapy once 19 hours after drug administration. A 630nm semiconductor laser was used, with an optical fiber equipped with a microlens at the end directly facing the target area to ensure the beam was perpendicular and completely covered the tumor. Parameters were set according to the following steps. Mice were anesthetized by inhalation using an isoflurane small animal anesthesia machine. After anesthesia, the mice were fixed with medical tape to fully expose the tumor site, and non-tumor areas were covered with black plastic bags.

[0181] a. Start the laser therapy device calibration and set the parameters.

[0182] b. When determining the laser treatment distance, the laser spot diameter should be able to cover the size of the tumor.

[0183] c. At the laser treatment distance given in the previous step, use an optical power meter to measure the laser power value per unit area.

[0184] d. Adjust the laser power value to 318mW.

[0185] e. Set the laser treatment time and perform PDT treatment on the experimental animals.

[0186] Table 10 Light Dosage Design Table

[0187] 4.7. Data Collection

[0188] After administration to the groups, the length and width of the tumors were measured twice a week using vernier calipers (purchased from Mitutoyo Precision Measuring Instruments (Shanghai) Co., Ltd.), and the mice were weighed simultaneously. 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: TV = length × width 2 / 2; TGI 体积 (%) = [1-(V) t -V0) 给药组 / (Vt -V0) 阴性对照组 ×100%;

[0189] Where V0 is the average tumor volume at the start of drug administration in a certain treatment group, V t The average tumor volume at the end of administration for this treatment group.

[0190] At the end of the experiment, mice in each group were euthanized, tumor tissue was harvested, weighed, and recorded. T / C was calculated. 重量 Percentage and tumor growth inhibition rate TGI 重量 (%). The specific calculation formula is as follows: T / C 重量 (%) = TW 给药组 / TW 阴性对照组 ×100%; TGI 重量 (%) = (1 - T / C) 重量 )×100%.

[0191] Among them, TW 给药组 The tumor weight in the treatment group is represented by TW. 阴性对照组 This represents the tumor weight in the negative control group.

[0192] 4.8. Sample Collection and Processing

[0193] At the end of the experiment, the mice in each group were euthanized, the tumor tissue was removed, and the tumor tissue was weighed.

[0194] 4.9. Data Processing and Statistical Analysis

[0195] All experimental results are expressed as mean ± standard error. GraphPad Prism 8 was used to plot the results, with the number of days after grouping on the x-axis and mouse body weight or tumor volume on the y-axis. Statistical analysis was performed based on the data obtained at the end of the experiment to assess differences between groups. Ordinary one-way ANOVA was used in GraphPad Prism 8 software for data analysis, and p < 0.05 was considered statistically significant.

[0196] 5. Results

[0197] weight change

[0198] During the experiment, the mice tolerated the test well, and the changes in body weight of mice in each group are shown in Figure 3 (data points represent the average body weight within the group).

[0199] Tumor volume

[0200] Table 11 shows the change in mean tumor volume over time in the A549 non-small cell lung cancer subcutaneous xenograft model.

[0201] Table 11 Tumor volume at different time points in each group Note: a. Mean ± standard error; b. Tumor growth curve and endpoint tumor weight after days of drug administration.

[0202] Tumor growth curves and tumor weights are shown in Figures 4 and 5. Data points represent the mean body weight within a group, and error bars represent the standard error (SEM).

[0203] Anti-tumor drug efficacy evaluation indicators

[0204] The growth inhibition rate of the test substance on the A549 non-small cell lung cancer subcutaneous xenograft model was calculated based on the tumor volume on day 22 after drug administration (Table 12). The tumor weight of all groups on day 22 is shown in Table 13.

[0205] Table 12 Growth inhibition rate of the test substances on the A549 non-small cell lung cancer subcutaneous xenograft model Note: a. Mean ± standard error; bp values ​​were analyzed using one-way ANOVA.

[0206] Table 13 Tumor weight and statistical analysis at the end of the experiment (day 22). Note: a. Mean ± standard error; bp values ​​were analyzed using one-way ANOVA.

[0207] 6. Conclusion

[0208] The above experiments evaluated the in vivo antitumor effects of sodium porphyrin and the targeted photosensitizer X-1 in a subcutaneous xenograft model of A549 non-small cell lung cancer. Animal body weight and tumor volume measured at different time points in each experimental group are shown in Figures 3 and 4, and the final tumor weight is shown in Figure 5.

[0209] The above experiments were conducted at an average tumor volume of 158 mm. 3 The subjects were divided into groups, and the drug was administered on the same day as their group assignment. Nineteen hours after drug administration, the subjects received a single exposure to light at a wavelength of 630 nm. The experiment concluded on day 22 after the group assignments. The results show that on day 22, the tumor volume in the negative control group reached 674 mmHg. 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 tumor growth in the A549 model. The positive control drug, sodium porphyrin, at a dose of 2 mg / kg (TV = 289 mmHg), showed no significant inhibitory effect. 3 The mitochondrial-targeting photosensitizer X-1 (TV = 169 mmHg, TGI = 74.7%, P < 0.001) significantly inhibited tumor growth in the A549 model. Compared with erlotinib and porphyrin sodium, the mitochondrial-targeting photosensitizer X-1 (TV = 169 mmHg) of this invention significantly inhibited tumor growth in the A549 model. 3The TGI (97.8%, P<0.001) showed better efficacy and small intragroup differences, demonstrating enhanced targeting ability and significantly improved efficacy.

Claims

1. A mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof, wherein the mitochondrial-targeting photosensitizer has the structure of the following general formula (I): Each of the groups R independently represents C. 3-9 Heterocyclic C 1-10 alkylene-O-, Wherein C 3-9 The heterocyclic group is unsubstituted or substituted by one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, alkylcarbonyl, haloalkyl or haloalkoxy; The C 1-10 The alkylene group is straight-chain or branched, unsubstituted, or substituted with one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, alkylcarbonyl, haloalkyl, or haloalkoxy, and the alkylene group may optionally be separated 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 heterocycles therein are unsubstituted or substituted by one or more of the same or different groups selected from the following: cyano, halogen, alkyl, alkoxy, alkylthio, cycloalkyl, haloalkyl or haloalkoxy.

2. The mitochondrial-targeting photosensitizer of claim 1 or a pharmaceutically acceptable salt thereof, wherein C 3-9 The heterocyclic group contains at least one nitrogen atom and is selected from the following groups: pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazolyl, dioxazolyl, isothiazolyl, pyridinyl, pyridinyl, pyrazinyl, triazinyl, tetraazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrolinyl, pyrrolylalkyl, imidazolinyl, imidazolyl, thiazolyl, oxazolylalkyl, pyrazolylalkyl, azacyclic butyl, azacyclic propyl, oxazcyclic butyl, oxazcyclic propyl, oxazcyclic heptyl, oxazcyclic hexyl, azacyclic heptyl, oxopyrrolyl, dioxopyrrolyl, oxomorpholinyl, and oxopiperazinyl.

3. The mitochondrial-targeting photosensitizer of claim 1 or a pharmaceutically acceptable salt thereof, wherein C 1-10 Alkylenes are straight-chain and unsubstituted.

4. The mitochondrial-targeting photosensitizer according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the group R is C 3-6 Heterocyclic 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, oxtriazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetraazinyl, oxazine, isoxazinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, pyrrololinyl, pyrrolylalkyl, imidazolinyl, imidazolyl, thiazolylalkyl, oxazolylalkyl, pyrazolylalkyl, azirrocyclobutyl, azirrocyclopropyl, oxazirrocyclobutyl, oxazirrocyclopropyl, oxazirrocyclohexyl, oxopyrrolylalkyl, dioxopyrrolylalkyl, oxomorpholinyl, and oxopyrazinyl; C 1-6 The alkyl group is selected from methylene, ethylene, propylene, n-butylene, n-pentylene, or n-hexylene.

5. The mitochondrial-targeting photosensitizer according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the group R is C 3-6 Heterocyclic C 1-4 alkylene-O-, where C 3-6 The heterocyclic group is selected from the following groups: pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, furazonyl, dioxazolyl, isothiazolyl, pyridazinyl, pyrazinyl, oxazine, isoxazinyl, piperazinyl, morpholinyl, thiomorpholinyl, imidazolinyl, imidazoalkyl, thiazoalkyl, oxazolalkyl, pyrazolalkyl, azirheptanyl, oxazirheptanyl, oxazirheptanyl, oxomorpholinyl, and oxopiperazinyl; C 1-4 The alkylene group is selected from methylene, ethylene, n-propylene, or n-butylene.

6. The mitochondrial-targeting photosensitizer of claim 1 or a pharmaceutically acceptable salt thereof, wherein the 3-6 member nitrogen-containing heterocyclic group is selected from the following groups: pyrrole, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, isoxazolyl, oxazolyl, oxadiazolyl, oxtriazolyl, furazolyl, dioxazolyl, isothiazolyl, pyridinyl, pyridinyl, piperidinyl, triazinyl, tetraazinyl, oxazine, isoxazinyl, or morpholinyl.

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

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-methylimidazolium, 1,8-diazabicyclo[5.4.0]undec-7-ene, triethylamine, pyridine, N,N'-dimethylaminopyridine, 2,6-dimethylpyridine, or mixtures thereof; The condensing agent is a urea cation type condensing agent, selected from O-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), O-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethylurea tetrafluoroborate (TATU), and O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroborate (T... BTU), O-(1,2-dihydro-2-oxo-pyridyl)-1,1,3,3-tetramethylurea tetrafluoroborate (TPTU); carbodiimide type condensing agents: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1,3-dicyclohexylcarbodiimide (DCC); or carbonylimidazolium type condensing agents: N,N-carbonyldiimidazole (CDI); or mixtures thereof, wherein 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 mixtures thereof.

9. The use of any one of claims 1 to 6, a mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for treating: cancer, including cancer of the reproductive organs, leukemia, mast cell tumor, breast cancer, kidney cancer, respiratory 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 said diseases.

10. A kit comprising the mitochondrial-targeting photosensitizer or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 6, in a therapeutically effective amount; and instructions for use therein in photodynamic therapy.

Images