Curcumin heterocoumarin photodiagnosis and treatment agent compounds, and preparation method and application thereof

Abstract

The application discloses a curcuminoid xanthene compound shown in formula I, a preparation method of the curcuminoid xanthene compound and application of the curcuminoid xanthene compound as a photodiagnosis and treatment agent. The compound has the characteristics of long absorption and emission wavelength, good stability, high phototoxicity, large Stokes shift, good biocompatibility and good cell membrane permeability. Compared with curcumin, the photosensitizer has a longer absorption wavelength, lower dark toxicity and can generate ROS under type-I and type-II synergistic action, and is expected to be applied to PDT in an anaerobic tumor microenvironment.

Description

Technical Field

[0001] This invention relates to a class of phototherapy agents. More specifically, it relates to a class of curcumin-coumarin phototherapy agent compounds, their preparation methods, and applications. Background Technology

[0002] Compared with traditional methods (surgery, radiotherapy, chemotherapy, antibiotics), photodynamic therapy (PDT) has the following advantages: non-invasive, high spatiotemporal precision, high selectivity for tumor destruction, repeatable treatment, and fewer toxic side effects. The therapeutic effect of PDT is achieved through a photosensitizer absorbing appropriate light energy to transition to a singlet excited state (S1), then through intersystem crossing to a triplet excited state (T1). T1 then transfers energy to O2 to generate singlet oxygen. 1 O2 (Type II reaction). Furthermore, T1 can also react with intracellular substrates (e.g., nucleic acids, proteins, and lipids) via electron transfer mechanisms to generate other ROS, such as hydroxyl radicals (OH·) and superoxide anions (O2). -· (Type I reaction). Reactive oxygen species (ROS) generated through two mechanisms damage biomolecules such as proteins, lipids, and nucleic acids, thereby harming tumor cells. Currently, phototherapy (PDT) is used for various superficial malignant lesions, vascular malformations, and bacterial infections. Reports have confirmed that photosensitizers can be used for fluorescence imaging during PDT, achieving an integrated diagnostic and therapeutic effect.

[0003] Hypoxia is a typical characteristic of the tumor microenvironment (TME), caused by the malignant proliferation of cancer cells and abnormal growth of tumor blood vessels. Common photosensitizers are usually type II photosensitizers, whose phototransformation treatment (PDT) effect is closely related to O2 levels. Furthermore, type II photosensitizers consume a large amount of O2 during PDT, further reducing O2 supply. Currently, type I photosensitizers have attracted widespread attention to improve PDT effects under hypoxic conditions. This type of photosensitizer, in the absence of oxygen, generates OH· and O2 via the type-I pathway after light irradiation. -· This kills tumor cells. Therefore, designing a class of type I and type II photosensitizers with synergistic effects is particularly important for photodynamic therapy under hypoxic conditions.

[0004] Curcumin, as a natural drug, possesses a variety of unique physiological and biological activities, such as antioxidant, anti-inflammatory, anticancer, antimicrobial, antibacterial, neuroprotective, and cardioprotective properties. However, its poor bioavailability, weak photodynamic effects, and instability in physiological media limit its further clinical application. Coumarins are heterocyclic compounds widely found in nature. Studies have shown that coumarins possess a wide range of biological activities, including antifungal, antibacterial, antiviral, anticancer, anticoagulant, and antihypertensive effects. Summary of the Invention

[0005] The first objective of this invention is to provide a curcumin-based phototherapy agent. This phototherapy agent has the characteristics of long absorption and emission wavelengths, good stability, ability to generate ROS through two pathways, good PDT effect, large Stokes shift, good biocompatibility, and good cell membrane permeability.

[0006] The second objective of this invention is to provide a method for preparing curcumin-coumarin phototherapy agents. This method is simple, based on readily available coumarin derivatives, and obtains a class of phototherapy agents with a curcumin-coumarin backbone through 2,4-pentanedione linkage.

[0007] The third objective of this invention is to provide an application of a curcumin-based phototherapy agent.

[0008] To achieve the first objective mentioned above, the present invention adopts the following technical solution:

[0009] This invention provides a compound represented by Formula I:

[0010]

[0011] Each of R1, R2, R3, and R4 may be the same or different, and is independently selected from hydrogen, halogen, CN, OH, NO2, unsubstituted, or optionally substituted by one, two, or more Rs. a The following groups are substituted: alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, NH2;

[0012] Each Y may be identical or different, and is independently selected from NR5R6 or OR7; or, Y forms unsubstituted or optionally substituted groups with one, two or more R atoms, respectively, with R1 and / or R4 and their respective attached atoms. c Substituted heterocycles;

[0013] R5, R6, and R7 may be the same or different, and are independently selected from hydrogen, unsubstituted, or optionally separated by one, two, or more R groups. b The following groups may be substituted: alkyl, alkenyl, alkynyl, alkyl-C(=O)-, cycloalkyl, aryl, heteroaryl, alkyl-NHC(=O)-alkyl-, (alkyl)2-NC(=O)-alkyl-, etheryl; for example, selected from -(CH2). m COOH, -(CH2) m COO(CH2) n CH3, -(CH2) m COOC(CH3)3、-(CH2) m CONH(CH2) n CH3, -(CH2) m CON[(CH2) nCH3]2、-(CH2) m SO3H, -(CH2) m OH, where m is 1-10 and n is 0-6; the number of carbon atoms in the ether group is 4-20 and the number of oxygen atoms is ≤8, for example selected from -CH2CH2OCH2CH3, -CH2CH2OCH2CH2OH, -CH2CH2(OCH2CH2)2CH2CH3, -CH2CH2(OCH2CH2)2CH2CH2OH, -CH2CH2(OCH2CH2)3CH2CH3, -CH2CH2(OCH2CH2)3CH2CH2OH, -CH2CH2(OCH2CH2)4CH2CH3 or -CH2CH2(OCH2CH2)4CH2CH2OH;

[0014] Each R a R b R c The same or different, independently selected from halogens, CN, OH, NO2, NH2, COOH, SO3H, alkyl, alkoxy, alkylC(=O)O-, alkoxy-C(=O)-, aryl, heteroaryl;

[0015] Furthermore, the compound represented by Formula I is not...

[0016] According to embodiments of the present invention, R1, R2, R3, and R4 may be the same or different, and are independently selected from hydrogen, halogens, CN, OH, NO2, and C. 1-20 Alkyl, C 2-20 alkenyl, C 2-20 alkynyl group, C 1-20 Alkoxy, C 3-20 Cycloalkyl groups, NH2.

[0017] According to embodiments of the present invention, R1, R2, R3, and R4 may be the same or different, and are independently selected from hydrogen, halogens, and C. 1-10 Alkyl, C 1-10 Alkyl group.

[0018] According to an embodiment of the present invention, when R3 is H, R5 and R6 are not both alkyl groups.

[0019] According to embodiments of the present invention, R5, R6, and R7 may be the same or different, and are independently selected from hydrogen, unsubstituted, or optionally replaced by one, two, or more Rs. b The following groups are substituted: C 1-20 Alkyl, C 2-20 alkenyl, C 2-20 alkynyl group, C 1-20 Alkyl-C(=O)-, C 3-20 cycloalkyl, C6-14 Aryl, 5-14 membered heteroaryl; or, R5 and R6 form 3-14 membered heterocyclic groups with the N attached to them;

[0020] According to an embodiment of the present invention, each R b They may be the same or different, and are selected independently from halogens, CN, OH, NO2, NH2, and C. 1-20 Alkyl, C 1-20 Alkoxy, C 1-20 Alkyl C(=O)O-, C 1-20 Alkoxy-C(=O)-, C 6-14 Aryl, 5-14 heteroaryl.

[0021] According to embodiments of the present invention, R5, R6, and R7 may be the same or different, and are independently selected from hydrogen, unsubstituted, or optionally replaced by one, two, or more Rs. b The following groups are substituted: C 1-10 Alkyl, C 1-10 Alkyl-C(=O)-;

[0022] According to an embodiment of the present invention, each R b Same or different, independent of each other, selected from C 1-10 Alkyl, C 1-10 Alkoxy, C 1-10 Alkoxy-C(=O)-, C 6-8 Aryl.

[0023] According to an embodiment of the present invention, R5, R6, and R7 may be the same or different, and are independently selected from hydrogen and C. 1-6 Alkyl, C 1-6 Alkyl-C(=O)-, C 6-8 Aryl C 1-6 Alkyl, C 1-6 Alkoxy C 1-6 Alkyl, C 1-10 Alkoxy-C(=O)-C 1-10 alkyl-.

[0024] According to embodiments of the present invention, R5, R6, and R7 may be the same or different, and are independently selected from H, methyl, ethyl, acetyl, propionyl, benzyl, ethoxymethyl, hydroxyethoxymethyl, and CH3CH2OC(=O)CH2-.

[0025] According to embodiments of the present invention, each Y may be the same or different, and is independently selected from NR5R6, OR7, or 3-14 membered heterocyclic groups; or, Y and R1 and / or R4 and the atoms respectively attached thereto form unsubstituted or optionally substituted groups with one, two or more R groups. cThe substituted 3-14 member heterocycle; the 3-14 member heterocycle is, for example, a 5-8 member heterocycle, such as a 6-membered N heterocycle;

[0026] According to an embodiment of the present invention, each Y may be the same or different, and is independently selected from OH, Alternatively, Y forms unsubstituted or optionally substituted atoms with R1 and / or R4 and their respective bonds. c The following structures are replaced:

[0027] Among them, R, R c Selected from H, halogens, C 1-6 Alkyl, C 1-6 Alkyl group; p is selected from integers from 0 to 8; Z is selected from NH, CH2, and O; For example,

[0028] According to an embodiment of the present invention, the compound represented by Formula I is selected from the following structures:

[0029]

[0030] The present invention also provides a method for preparing the compound shown in Formula I, comprising the following steps: reacting compound II with compound 1 (2,4-pentanedione) to obtain the compound shown in Formula I;

[0031]

[0032] Among them, R1, R2, R3, R4, and Y have the definitions described above.

[0033] According to an embodiment of the present invention, when the compound shown in Formula I has a symmetrical structure, the compound shown in Formula I can be obtained in one step; when the compound shown in Formula I has an asymmetrical structure, those skilled in the art should know that the compound shown in Formula I can be obtained stepwise by adjusting the type and amount of compound II.

[0034] According to an embodiment of the present invention, the reaction can be carried out in the presence of a base, the base being selected from organic bases, such as at least one selected from 1,2,3,4-tetrahydroisoquinoline, triethylamine, piperidine, diisopropylethylamine, and DMAP;

[0035] According to an embodiment of the present invention, the process can be carried out in a solvent selected from at least one of acetonitrile, N,N-dimethylformamide, and tetrahydrofuran.

[0036] This invention also provides intermediate compounds represented by Formula II:

[0037]

[0038] Among them, R1, R2, R3, R4, and Y have the definitions described above.

[0039] According to an embodiment of the present invention, the compound represented by Formula II is selected from the following structures:

[0040]

[0041]

[0042] The present invention also provides a pharmaceutical composition comprising the compound of formula I described herein.

[0043] According to an embodiment of the present invention, the pharmaceutical composition is a phototherapy agent.

[0044] This invention also provides the use of the compound of Formula I in the preparation of photodynamic drugs. This invention further provides the use of the compound of Formula I in the preparation of phototherapeutic agents for photodynamic diagnosis and treatment. The phototherapeutic agents are preferably used for the diagnosis and treatment of cancer at the cellular level or in vivo.

[0045] This invention also provides the application of compound I-2 in the preparation of photodynamic drugs.

[0046]

[0047] Beneficial effects

[0048] This invention provides a compound of Formula I, which is a type of compound I based on curcumin-coumarin backbone. It organically combines curcumin and coumarin dyes, and by introducing coumarin to increase the conjugated system, it causes a red shift in wavelength. Under illumination, the compound simultaneously generates O2 through both type-I and type-II pathways. -· and 1 Both O2 and the two ROS possess strong photodynamic effects, achieving not only good tumor cell killing effects but also promising applications in hypoxic tumor microenvironment phototherapy (PDT). Furthermore, their large Stokes shift avoids interference from autofluorescence, making them potential candidates for fluorescent probes. Simultaneously, these phototherapeutic agents exhibit good biocompatibility and high cell membrane permeability. Compared to curcumin, their absorption wavelength exhibits a redshift, further improving biocompatibility. They can also generate ROS through the synergistic action of type-I and type-II ROS, mitigating some of the shortcomings of curcumin. This allows for integrated diagnosis and treatment during therapy, holding profound significance for life science research.

[0049] The compounds of this invention generate reactive oxygen species (ROS) under illumination through the synergistic action of Type I and Type II, thereby killing tumor cells. These compounds not only possess a large Stokes shift but also a high fluorescence quantum yield, enabling them to achieve integrated diagnostic and therapeutic effects as phototherapy agents. Attached Figure Description

[0050] Figure 1 Absorption spectra of curcumin (DCM), compounds I-1 (MeOH), I-2 (DCM), and I-3 (DCM).

[0051] Figure 2 Fluorescence spectra of curcumin (DCM), compounds I-1 (MeOH), I-2 (DCM), and I-3 (DCM).

[0052] Figure 3 Solution method for detecting the formation of O2 from compound I-1 -· .

[0053] Figure 4 Solution method for detecting the formation of O2 from compound I-2 -· .

[0054] Figure 5 Electron spin resonance spectroscopy (ESR) was used to detect the generation of O2 from compound I-3. -· .

[0055] Figure 6 ESR detection of compound I-4 generating O2 -· .

[0056] Figure 7 ESR detection of O2 generation from compound I-6 -· .

[0057] Figure 8 ESR detection of O2 generation from compound I-8 -· .

[0058] Figure 9 ESR detection of O2 generation from compound I-9 -· .

[0059] Figure 10 ESR detection of O2 generation from compound I-11 -· .

[0060] Figure 11 ESR detection of compound I-3 formation 1 O2.

[0061] Figure 12 ESR detection of compound I-5 formation 1 O2.

[0062] Figure 13 ESR detection of compound I-7 formation 1 O2.

[0063] Figure 14 ESR detection of compound I-10 formation 1 O2.

[0064] Figure 15 ESR detection of compound I-12 formation 1 O2.

[0065] Figure 16 Compound I-1 was studied for its cytotoxicity to HepG-2 cells under dark conditions, phototoxicity, and phototoxicity under hypoxic conditions.

[0066] Figure 17 Compound I-2 was studied for its cytotoxicity to HepG-2 cells under dark conditions, phototoxicity, and phototoxicity under hypoxic conditions.

[0067] Figure 18 Compound I-3 exhibits cytotoxicity and phototoxicity in 4T1 cells.

[0068] Figure 19 Compound I-4 exhibits cytotoxicity and phototoxicity to HepG-2 cells.

[0069] Figure 20 Compound I-8 exhibits cytotoxicity and phototoxicity to HepG-2 cells.

[0070] Figure 21 Compound I-9 exhibits cytotoxicity and phototoxicity in HeLa cells.

[0071] Figure 22 Compound I-10 exhibits cytotoxicity and phototoxicity in HeLa cells.

[0072] Figure 23 Compound I-11 exhibits cytotoxicity and phototoxicity in HeLa cells.

[0073] Terminology Definitions and Explanations

[0074] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should be understood as being within the scope of this application specification and / or claims.

[0075] Unless otherwise stated, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-40" is equivalent to describing each integer value in the numerical range "1-10", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and each integer value in the numerical range "11-40", namely 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40. Furthermore, when certain numerical ranges are defined as "numbers", it should be understood that they describe the two endpoints of the range, each integer within the range, and each decimal within the range. For example, "numbers from 0 to 10" should be understood as not only recording each integer of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also recording at least the sum of each of these integers with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8 and 0.9 respectively.

[0076] It should be understood that in this article, when describing one, two or more, "more" should refer to integers greater than 2, such as 3 or greater than or equal to 3, such as 3, 4, 5, 6, 7, 8, 9 or 10.

[0077] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0078] The term "alkyl" should be understood to refer to a straight-chain or branched saturated monovalent hydrocarbon group having 1-40 carbon atoms. For example, "C 1-10 "Alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. 1-8 "Alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1-6 "Alkyl" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers.

[0079] The term "alkenyl" should be understood to refer to a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2-40 carbon atoms, preferably "C". 2-10 "Alkenyl". "C" 2-10 "Alkenyl" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, more preferably "C 2-8 "Alkenyl". "C" 2-10 "Alkenyl" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7 or 8 carbon atoms, for example, having 2, 3, 4, 5 or 6 carbon atoms (i.e., C... 2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C24, C34, C4 ... 2-3 Alkenyl). It should be understood that when the alkenyl group contains more than one double bond, the double bonds may be separable or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)- Pentyl-1-enyl, (Z)-pentyl-1-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl 2-Methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl But-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0080] The term "alkynyl" should be understood to refer to a straight or branched monovalent hydrocarbon group containing one or more triple bonds and having 2 to 40 carbon atoms, preferably "C". 2-10 "Alkyne group". The term "C" 2-10"Alkyne" should be understood to preferably represent a straight or branched monovalent hydrocarbon group containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, for example, having 2, 3, 4, 5, 6, 7, or 8 carbon atoms (i.e., "C"). 2-8 "Alkyne group" has 2, 3, 4, 5 or 6 carbon atoms (i.e., "C"). 2-6 The alkynyl group ("C") has 2 or 3 carbon atoms ("C") 2-3 The alkynyl group is, for example, ethynyl, prop-1-alkynyl, prop-2-alkynyl, but-1-alkynyl, but-2-alkynyl, but-3-alkynyl, pent-1-alkynyl, pent-2-alkynyl, pent-3-alkynyl, pent-4-alkynyl, hex-1-alkynyl, hex-2-alkynyl, hex-3-alkynyl, hex-4-alkynyl, hex-5-alkynyl, 1-methylprop-2-alkynyl, 2-methylbut-3-alkynyl, 1-methylbut-3-alkynyl, 1-methylbut-2-alkynyl, 3-methylbut-1-alkynyl, 1-ethylprop-2-alkynyl, 3-methylpent-4-alkynyl, 2-methylpent-4-alkynyl, 1-methylpent-4-alkynyl -Alynyl, 2-methylpentan-3-ynyl, 1-methylpentan-3-ynyl, 4-methylpentan-2-ynyl, 1-methylpentan-2-ynyl, 4-methylpentan-1-ynyl, 3-methylpentan-1-ynyl, 2-ethylbutan-3-ynyl, 1-ethylbutan-3-ynyl, 1-ethylbutan-2-ynyl, 1-propylpropan-2-ynyl, 1-isopropylpropan-2-ynyl, 2,2-dimethylbutan-3-ynyl, 1,1-dimethylbutan-3-ynyl, 1,1-dimethylbutan-2-ynyl, or 3,3-dimethylbutan-1-ynyl. In particular, the ynyl group is ethynyl, propan-1-ynyl, or propan-2-ynyl.

[0081] The term "cycloalkyl" should be understood to refer to saturated monocyclic, bicyclic (e.g., fused, bridged, spirocyclic) or tricyclic alkanes having 3 to 40 carbon atoms, preferably "C". 3-10 "Cycloalkyl", more preferably "C" 3-8 cycloalkyl. The term "C" 3-10 "Cycloalkyl" should be understood to refer to a saturated monovalent monocyclic, bicyclic (e.g., bridged, spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The C... 3-10Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl.

[0082] Unless otherwise defined, the term "heterocyclic group" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6-, or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11-, or 12-membered bicyclic ring (such as a fused ring, bridged ring, or spirocyclic ring), or a 10-, 11-, 12-, 13-, 14-, or 15-membered tricyclic ring system, and contains at least one, for example, 1, 2, 3, 4, 5, or more heteroatoms selected from O, S, and N, wherein N and S may optionally be oxidized to various oxidation states to form nitrides, -S(O)-, or -S(O)2- states. Preferably, the heterocyclic group may be selected from "3- to 10-membered heterocyclic groups". The term "3- to 10-membered heterocyclic group" means a saturated or unsaturated non-aromatic ring or ring system containing at least one heteroatom selected from O, S, and N. The heterocyclic group can be connected to the rest of the molecule via any one of the carbon atoms or a nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. Specifically, the heterocyclic group can include, but is not limited to: 4-membered rings, such as azirrobutyl or oxobutyl; 5-membered rings, such as tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoalkyl, pyrazolealkyl, or pyrrololinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazineyl, or trithiaalkyl; or 7-membered rings, such as diazacycloheptyl. Optionally, the heterocyclic group can be benzofused. The heterocyclic group can be bicyclic, such as, but not limited to, a 5,5-membered ring, like a hexahydrocyclopentano[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, like a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclic group can be partially unsaturated, meaning it can contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrroleyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl, or 4H-[1,4]thiazinyl, or it can be benzofused, such as, but not limited to, dihydroisoquinolinyl. When the 3-20-membered heterocyclic group is linked to other groups to form the compounds of the present invention, the carbon atom on the 3-20-membered heterocyclic group can be linked to other groups, or the heterocyclic atom on the 3-20-membered heterocyclic ring can be linked to other groups. For example, when the 3-20 membered heterocyclic group is selected from piperazine, the nitrogen atom on the piperazine group can be attached to other groups. Or when the 3-20 membered heterocyclic group is selected from piperidinium, the nitrogen atom on the piperidinium ring and the carbon atom at its para position can be attached to other groups.

[0083] The term "aryl" should preferably be understood to refer to a monocyclic, bicyclic (e.g., fused, bridged, or spirocyclic), or tricyclic hydrocarbon ring having 6 to 20 carbon atoms, which can be a monoaromatic ring or a polyaromatic ring fused together. Preferably, "C" indicates a single aromatic ring or a partially aromatic ring. 6-14 "Aromatic". The term "C" 6-14"Aryl" should be understood to preferably represent a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms. 6-14 Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, or rings with 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups, or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. When the C 6-20 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0084] The term "heteroaryl" should be understood to include monocyclic, bicyclic (e.g., fused, bridged, spirocyclic), or tricyclic aromatic ring systems having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O, and S, for example, "5-14-membered heteroaryl". The term "5-14-membered heteroaryl" should also be understood to include monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S, and in each case, may be benzo-fused. When the 5-20-membered heteroaryl is linked with other groups to form the compounds of the present invention, the linkage can be between carbon atoms on the 5-20-membered heteroaryl ring and other groups, or between heteroatoms on the 5-20-membered heteroaryl ring and other groups. When the 5-20 membered heteroaryl groups are substituted, they can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution sites; for example, hydrogen atoms bonded to carbon atoms on the heteroaryl ring can be substituted, or hydrogen atoms bonded to heteroatoms on the heteroaryl ring can be substituted.

[0085] The term "spirocycle" refers to a ring system in which two rings share a single ring atom.

[0086] The term "fused ring" refers to a ring system in which two rings share two cyclic atoms.

[0087] The term "bridged ring" refers to a ring system in which two rings share three or more cyclic atoms.

[0088] Unless otherwise stated, heterocyclic, heteroaryl, or heteroaryl groups include all possible isomers, such as their positional isomers. Thus, for some illustrative, non-limiting examples, forms may include those in which one, two, or more of the following positions (if present) are substituted or bonded to other groups, including pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thiophene or thiophene groups include thiophene-2-yl, thiophene-2-yl, thiophene-3-yl, and thiophene-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0089] The term "alkoxy" refers to -O-(alkyl), where alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, and butoxy. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, or heterocycloalkyloxy.

[0090] "Halogenated alkyl" refers to an alkyl group that has been substituted with one or more halogens, wherein the alkyl group is as defined above. Detailed Implementation

[0091] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0092] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0093] Example 1:

[0094]

[0095] 7-Hydroxycoumarin-3-carboxaldehyde II-1 was prepared from 2,4-dihydroxybenzaldehyde according to the literature (Chem. Commun., 2017, 53, 9470), with an overall yield of 25%.

[0096] 0.76 g (4 mmol) of compound II-1, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 18 mL of N,N-dimethylformamide to form a mixed solution. 0.08 mL (0.6 mmol) of triethylamine was added with stirring, and the mixture was reacted under reflux for 5 hours. After cooling, 100 mL of water was added, and the mixture was extracted three times with 200 mL of ethyl acetate. The ethyl acetate phase was dried and purified by column chromatography to obtain compound I-1 in 5% yield. ESIMS: m / z, 444.1. ab. max / nm=440nm,λ em max / nm=550nm, stokes=110nm, singlet oxygen yield Ф=0.12.

[0097] Example 2:

[0098]

[0099] 7-(diethylamino)coumarin-3-carboxaldehyde II-2 was prepared from 4-N,N-diethylaminosalicylic aldehyde according to the literature (Tetrahedron 68 (2012) 5458-5463), with an overall yield of 60%.

[0100] 0.49 g (2 mmol) of compound II-2, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 20 mL of acetonitrile to form a mixed solution. 0.1 mL (0.75 mmol) of 1,2,3,4-tetrahydroisoquinoline was added with stirring, and the mixture was reacted under reflux for 2 hours. After cooling, 100 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-2 was purified by column chromatography in 53.4% ​​yield. ESI MS: m / z, 554.2. ab. max / nm=525nm,λ em max / nm=600nm,Ф f =0.69, stokes = 75nm, singlet oxygen yield Ф = 0.27.

[0101] Example 3:

[0102]

[0103] II-3 was prepared from 2,4-dihydroxybenzaldehyde according to the literature (Anal. Chem. 2020, 92, 10068-10075), with an overall yield of 10%.

[0104] 1.08 g (4 mmol) of compound II-3, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 30 mL of N,N-dimethylformamide to form a mixed solution. 0.12 mL (0.9 mmol) of triethylamine was added with stirring, and the mixture was reacted under reflux for 12 hours. After cooling, 100 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-3 was purified by column chromatography in 8% yield. ESI MS: m / z, 602.2. ab. max / nm=545nm,λ em max / nm=620nm, stokes=75nm, singlet oxygen yield Ф=0.23.

[0105] Example 4:

[0106]

[0107] II-4 was prepared from 3-aminophenol according to the literature (Org. Lett. 2008, 10, 21, 5015–5018), with an overall yield of 15%.

[0108] 0.6 g (2 mmol) of compound II-4, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 25 mL of tetrahydrofuran to form a mixed solution. 0.03 mL (0.6 mmol) of piperidine was added with stirring, and the mixture was reacted under reflux for 8 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-4 was purified by column chromatography in 23% yield. ESI MS: m / z, 674.3. ab. max / nm=540nm,λ em max / nm=615nm, stokes=75nm, singlet oxygen yield Ф=0.31.

[0109] Example 5:

[0110]

[0111] II-5 was prepared from 3-aminophenol according to the literature (Org. Lett. 2008, 10, 21, 5015–5018), with an overall yield of 17%.

[0112] 0.9 g (3 mmol) of compound II-5, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 40 mL of acetonitrile to form a mixed solution. 0.06 mL (0.45 mmol) of triethylamine was added with stirring, and the mixture was reacted under reflux for 4 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-5 was purified by column chromatography in 33% yield. ESI MS: m / z, 610.2. ab. max / nm=505nm,λ em max / nm=570nm, stokes=65nm, singlet oxygen yield Ф=0.29.

[0113] Example 6:

[0114]

[0115] II-6 was prepared from 7-amino-4-methylcoumarin according to the literature (Zhurnal Obshchei Khimii, 1992, 62(7), 1658, 1664;), with an overall yield of 40%.

[0116] 0.55 g (2 mmol) of compound II-6, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 30 mL of N,N-dimethylformamide to form a mixed solution. 0.12 mL (0.9 mmol) of 1,2,3,4-tetrahydroisoquinoline was added with stirring, and the mixture was reacted under reflux for 12 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-6 was purified by column chromatography in 42% yield. ESI MS: m / z, 610.2. ab. max / nm=515nm,λ em max / nm=590nm, stokes=75nm, singlet oxygen yield Ф=0.27.

[0117] Example 7:

[0118]

[0119] II-7 was prepared from 7-diethylamino-4-methylcoumarin according to the literature (Anal. Chem. 2020, 92, 10068-10075), with an overall yield of 56%.

[0120] 0.52 g (2 mmol) of compound II-7, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 20 mL of acetonitrile to form a mixed solution. 0.08 mL (0.6 mmol) of 1,2,3,4-tetrahydroisoquinoline was added with stirring, and the mixture was reacted under reflux for 6 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-7 was purified by column chromatography in 47% yield. ESI MS: m / z, 582.3. ab. max / nm=530nm,λ em max / nm=585nm, stokes=55nm, singlet oxygen yield Ф=0.25.

[0121] Example 8:

[0122]

[0123] II-8 was prepared from 3-aminophenol according to the literature (Org. Lett. 2008, 10, 21, 5015–5018), with an overall yield of 10%.

[0124] 1.34 g (4 mmol) of compound II-8, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 35 mL of tetrahydrofuran to form a mixed solution. 0.1 mL (1 mmol) of piperidine was added with stirring, and the mixture was reacted under reflux for 9 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-8 was purified by column chromatography in 39% yield. ESI MS: m / z, 738.3. ab. max / nm=540nm,λ em max / nm=600nm, stokes=60nm, singlet oxygen yield Ф=0.29.

[0125] Example 9:

[0126]

[0127] II-9 was prepared from 3-N,N-diethylaminophenol according to the literature (Biosensors and Bioelectronics 90 (2017) 117–124), with an overall yield of 60%.

[0128] 0.56 g (2 mmol) of compound II-9, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 40 mL of acetonitrile to form a mixed solution. 0.1 mL (0.75 mmol) of triethylamine was added with stirring, and the mixture was reacted under reflux for 8 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-9 was purified by column chromatography in 21% yield. ESI MS: m / z, 622.2. ab. max / nm=528nm,λ em max / nm=600nm, stokes=72nm, singlet oxygen yield Ф=0.20.

[0129] Example 10:

[0130]

[0131] II-10 was prepared from 3-aminophenol according to the literature (Org. Lett. 2008, 10, 21, 5015–5018), with an overall yield of 5%.

[0132] 0.72 g (2 mmol) of compound II-10, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 30 mL of N,N-dimethylformamide to form a mixed solution. 0.1 mL (0.75 mmol) of 1,2,3,4-tetrahydroisoquinoline was added with stirring, and the mixture was reacted under reflux for 5 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-10 was purified by column chromatography in 55% yield. ESI MS: m / z, 786.2. ab. max / nm=510nm,λ em max / nm=580nm, stokes=70nm, singlet oxygen yield Ф=0.27.

[0133] Example 11:

[0134]

[0135] II-11 was prepared from 3-bromophenol according to the literature (Bioorganic & Medicinal Chemistry Letters 27(2017)4893–4897), with an overall yield of 16%.

[0136] 1.2 g (4 mmol) of compound II-4, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 20 mL of N,N-dimethylformamide to form a mixed solution. 0.1 mL (1 mmol) of piperidine was added with stirring, and the mixture was reacted under reflux for 10 hours. After cooling, 200 mL of water was added, and the mixture was extracted three times with 200 mL of dichloromethane. After drying the dichloromethane phase, compound I-11 was purified by column chromatography in 23% yield. ESI MS: m / z, 650.2. ab. max / nm=535nm,λ em max / nm=610nm, stokes=75nm, singlet oxygen yield Ф=0.22.

[0137] Example 12:

[0138]

[0139] II-12 was prepared from psoralen according to the literature (Tetrahedron 68 (2012) 5458-5463), with a total yield of 6%.

[0140] 0.63 g (3 mmol) of compound II-12, 0.11 mL (1 mmol) of 2,4-pentanedione, 0.69 g (10 mmol) of boron trioxide, and 2.3 g (10 mmol) of tributyl borate were mixed in 30 mL of tetrahydrofuran to form a mixed solution. 0.08 mL (0.6 mmol) of 1,2,3,4-tetrahydroisoquinoline was added with stirring, and the mixture was reacted under reflux for 5 hours. After cooling, 100 mL of water was added, and the mixture was extracted three times with 200 mL of ethyl acetate. The ethyl acetate phase was dried and purified by column chromatography to obtain compound I-12 in 14% yield. ESI MS: m / z, 492.1. ab. max / nm=460nm,λ em max / nm=535nm, stokes=75nm, singlet oxygen yield Ф=0.17.

[0141] Test Example 1

[0142] Cytotoxicity was assessed using the standard MTT assay.

[0143] Dark cytotoxicity assay: First, HeLa cells were seeded into 96-well plates and incubated with different concentrations of I-2 solution at 37°C and 5% CO2 for 1 hour. Then, the culture medium was replaced with fresh medium, and incubation continued for 24 hours. 20 μL of MTT solution (5 mg / mL) was added to each well. -1 Continue incubation for 4 hours, remove the solution from each well, add 80 μL of DMSO solution, and measure the absorbance at 570 nm using a microplate reader. Using the absorbance value of the control group as a reference, and assuming its survival rate is 100%, calculate the cell viability for different concentrations. Each group of cells was measured in parallel 6 times.

[0144] Cellular PDT assay: First, HeLa cells were seeded into 96-well plates and incubated with different concentrations of I-2 solution at 37°C and 5% CO2 for 1 hour. Then, the culture medium was replaced with fresh medium, and the cells were incubated with a 532nm laser (0.1W cm⁻¹). -2 Irradiate for 10 minutes, then continue incubation, replacing with fresh culture medium and continuing incubation for 24 hours. Add 20 μL of MTT solution (5 mg / mL) to each well. -1 Continue incubation for 4 hours, remove the solution from each well, add 80 μL of DMSO solution, and measure the absorbance at 570 nm using a microplate reader. Using the absorbance value of the control group as a reference, and assuming its survival rate is 100%, calculate the cell viability for different concentrations. Each group of cells was measured in parallel 6 times.

[0145] Figures 3-15 This indicates that the compounds of the present invention, as photosensitizers, can effectively generate singlet oxygen (…). 1 O2) and superoxide anion (O2) -· This allows for the application of phototherapy. Figure 16-23 The results show that the compounds of the present invention have low dark toxicity and extremely high phototoxicity, and possess advantages such as non-invasiveness, high spatiotemporal precision, high tumor destruction selectivity, repeatable treatment, and few toxic side effects.

[0146] The embodiments of the technical solution of the present invention have been described above by way of example. It should be understood that the protection scope of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the protection scope of the claims of this application.

Claims

1. A compound represented by Formula I: Ⅰ Each of R1, R2, and R4 is hydrogen; R3 is selected from hydrogen, halogens, and C. 1-6 alkyl; Each Y may be the same or different, and each can be selected independently from -NR5R6 or -OR7; R5 and R6 may be the same or different, and are independently selected from hydrogen and C. 1-6 Alkyl, C 1-6 Alkyl-C(=O)-, C 6-8 Aryl C 1-6 Alkyl, C 1-6 Alkoxy C 1-6 Alkyl, C 1-6 Alkoxy-C(=O)-C 1-6 alkyl; R7 is selected from hydrogen, C 1-6 alkyl; And the compound represented by Formula I is not .

2. The compound according to claim 1, characterized in that, When R3 is H, R5 and R6 are not simultaneously C. 1-6 alkyl.

3. The compound according to claim 1, characterized in that, R5 and R6 may be the same or different, and are independently selected from H, methyl, ethyl, acetyl, propionyl, benzyl, ethoxymethyl, CH3CH2OC(=O)CH2-; R7 is selected from H, methyl, and ethyl.

4. The compound according to claim 1, characterized in that, Each Y may be the same or different, and each is independently selected from -OH, , , , , , , .

5. A compound, characterized in that, Selected from the following structure: 。 6. A method for preparing the compound according to any one of claims 1-5, comprising the following steps: Compound II reacts with 2,4-pentanedione to give the compound shown in Formula I; Wherein, R1, R2, R3, R4, and Y have the definitions described in any one of claims 1-5; And / or, the reaction is carried out in the presence of a base selected from at least one of 1,2,3,4-tetrahydroisoquinoline, triethylamine, piperidine, diisopropylethylamine, and DMAP; And / or, the reaction is carried out in a solvent selected from at least one of acetonitrile, N,N-dimethylformamide, and tetrahydrofuran.

7. The use of the compound according to any one of claims 1-5 in the preparation of a phototherapy agent, said phototherapy agent being used for the diagnosis and treatment of tumors at the cellular level or in vivo.

8. A photodynamic pharmaceutical composition comprising the compound according to any one of claims 1-5.

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