Near-infrared xanthene dye with adjustable structural symmetry and preparation method and application thereof

Abstract

This invention belongs to the field of biomedical technology, specifically relating to a near-infrared oxanthracene dye with tunable structural symmetry, its preparation method, and its application. Its structural formula is shown in formula Ia or Ib. This dye exhibits high quantum yield (up to 79.5%), low dark toxicity and phototoxicity, or, under near-infrared laser irradiation, can effectively produce type I (… 1 O2) and type II (•OH, O2• ‑ Reactive oxygen species can enable precise visualization imaging of mitochondria and achieve highly effective treatment of tumor cells by inducing ferroptosis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a near-infrared oxanthracene dye with tunable structural symmetry, its preparation method, and its application, the application being in imaging-mediated photodynamic therapy. Background Technology

[0002] The statements herein provide only background information in relation to this invention and do not necessarily constitute prior art.

[0003] Fluorescence imaging technology, with its advantages of real-time performance, speed, non-destructive nature, and high sensitivity, has seen rapid development in fields such as food safety, environmental monitoring, and biomedicine. Small molecule fluorescent dyes, as one of the most important tools in fluorescence imaging, play a crucial role in studying the relationship between substances related to life activities and the physiological and pathological processes of diseases. However, the requirements for dye molecule properties vary across different application areas.

[0004] Photodynamic therapy (PDT), a minimally invasive treatment method, utilizes photosensitive dyes combined with light to treat cancer. Upon activation, its triplet excited state can undergo a photochemical reaction in the presence of oxygen, producing reactive oxygen species (ROS), including type II ROS (Reactive Oxygen Species). 1 O2) and type I ROS (O2) • (and •OH). These ROS effectively disrupt biomolecules and inhibit cancer growth by triggering various cell death pathways (such as apoptosis, ferroptosis, etc.). Therefore, developing dye molecular backbones that can be applied in multiple fields through simple regulatory strategies is of great significance.

[0005] Oxanthracene dyes possess excellent photophysical properties such as large molar extinction coefficients and high fluorescence quantum yields. Furthermore, they have numerous modifiable sites, making them widely used in bioimaging, fluorescence detection, and tumor therapy. To optimize the performance of oxanthracene dyes for various applications, researchers have developed three main strategies: (1) introducing electron-withdrawing or electron-donating groups into the oxanthracene skeleton; (2) increasing the rigidity of the oxanthracene structure; and (3) replacing the oxygen atom at position 10 with other main group elements (S, Si, P, etc.). However, the compounds designed and reported based on these strategies often involve cumbersome synthetic steps, large molecular weights, and complex molecular structures, and the same molecular skeleton is often only applicable to a single application.

[0006] However, according to literature review, there are currently no reports of oxanthracene dyes that can simultaneously meet the application needs of multiple fields through simple structural modulation. Exploration in this direction still has important scientific research significance and application value. Summary of the Invention

[0007] To address the shortcomings of existing technologies, the purpose of this invention is to provide a near-infrared oxanthracene dye with tunable structural symmetry, its preparation method, and its application.

[0008] To achieve the above objectives, the present invention is implemented through the following technical solution:

[0009] In a first aspect, the present invention provides a near-infrared oxanthracene dye with tunable structural symmetry, the structural formula of which is shown in formula Ia or Ib: ; Among them, R1, R2, R5 to R 12 It is independently selected from hydrogen, C1-C6 straight-chain hydrocarbon group, C1-C6 branched hydrocarbon group, C1-C6 alkoxy group, carboxylic acid ester group, benzene ring, and any one of C1-C6 alkyl, alkoxy, substituted thiophene and substituted benzene ring optionally substituted with C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkynyl, C1-C4 alkynyloxy, ethyl acetate group, cyano, trifluoromethyl and halogen substituted C1-C6 alkyl, alkoxy, substituted thiophene and substituted benzene ring; R3 and R4 are independently selected from any one of hydrogen, C1-C6 alkyl, fluorine, chlorine, bromine, iodine, cyano, trifluoromethyl, and C1-C6 alkoxy. X1 and X2 are independently selected , , , , , , , , , , , , one of the; Z - Independently selected from halide ions, ClO4 - PF6 - BPh4 - N(CN)2 - and BF4 - Any one of them.

[0010] In some embodiments, R1 and R2 are independently selected from hydrogen and C1-C6 straight-chain alkyl groups.

[0011] Preferably, R1 and R2 are independently selected from C1-C3 straight-chain alkyl groups.

[0012] Secondly, the present invention provides a method for preparing the benzene ring-modified near-infrared oxanthracene dye, the synthetic route of which is as follows: ; Compound 1 and Compound 2 were added to HClO4 in a certain proportion and heated to react at a temperature of 90-110℃ for 16-20 h. After the reaction was completed, the mixture was cooled and an aqueous solution of Z salt or Z acid was added. After purification, the product was obtained. Z - Independently selected from halide ions, ClO4 - PF6 - BPh4 - N(CN)2 - and BF4 - Any one of them.

[0013] In some embodiments, the molar ratio of compound 1 to compound 2 is 1.5-2.5:1. For example, it can be 1.5:1, 1.6:1, 1.7:1, 1.8:1, 1.9:1, 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1, or 2.5:1.

[0014] Preferably, the molar ratio of compound 1 to compound 2 is 1.8-2.3:1.

[0015] More preferably, the molar ratio of compound 1 to compound 2 is 2:1.

[0016] In some embodiments, the reaction temperature of compound 1 and compound 2 is 95-110°C, preferably 100-110°C, and more preferably 100-105°C. For example, the reaction temperature can be 95°C, 96°C, 97°C, 98°C, 99°C, 100°C, 101°C, 102°C, 103°C, 104°C, 105°C, 106°C, 107°C, 108°C, 109°C, or 110°C.

[0017] In some embodiments, the concentration of compound 2 in HClO4 is 0.1-1 mol / L. For example, it can be 0.1 mol / L, 0.2 mol / L, 0.3 mol / L, 0.4 mol / L, 0.5 mol / L, 0.6 mol / L, 0.7 mol / L, 0.8 mol / L, 0.9 mol / L, or 1 mol / L.

[0018] In some embodiments, the purification method is a silica gel column separation and purification method.

[0019] Preferably, the eluent used for silica gel column separation and purification is a mixture of dichloromethane and ethanol, with a volume ratio of dichloromethane to ethanol of 20-100:1.

[0020] In some embodiments, the molar ratio of Z salt or Z acid to compound 2 is 3-6:1. For example, it can be 3:1, 4:1, 5:1 or 6:1.

[0021] Thirdly, the present invention provides a method for preparing the heterocyclic modified near-infrared oxanthracene dye, the synthetic route of which is as follows: ; Compound 1 and compound 3 were added to HClO4 in a certain proportion and heated to react at a temperature of 90-110 °C for 16-20 h. After the reaction was completed, the mixture was cooled and an aqueous solution of Z salt or Z acid was added. After purification, the product was obtained.

[0022] In some embodiments, the molar ratio of compound 1 to compound 3 is 1.5-2.5:1.

[0023] Preferably, the molar ratio of compound 1 to compound 3 is 1.8-2.3:1.

[0024] More preferably, the molar ratio of compound 1 to compound 3 is 2:1.

[0025] In some embodiments, the concentration of compound 3 in HClO4 is 0.1-1 mol / L. For example, it can be 0.1 mol / L, 0.2 mol / L, 0.3 mol / L, 0.4 mol / L, 0.5 mol / L, 0.6 mol / L, 0.7 mol / L, 0.8 mol / L, 0.9 mol / L, or 1 mol / L.

[0026] In some embodiments, the purification method is a silica gel column separation and purification method.

[0027] Preferably, the eluent used for silica gel column separation and purification is a mixture of dichloromethane and ethanol, with a volume ratio of dichloromethane to ethanol of 20-100:1.

[0028] In some embodiments, the molar ratio of Z salt or Z acid to compound 3 is 3-6:1. For example, it can be 3:1, 4:1, 5:1 or 6:1.

[0029] Fourthly, the present invention provides the application of the benzene ring / heterocyclic modified near-infrared xanthracene dyes in mitochondrial imaging, preparation of anticancer drugs, and photodynamic therapy.

[0030] The beneficial effects achieved by one or more embodiments of the present invention described above are as follows: (1) This invention can be synthesized through a simple nucleophilic reaction. By substituting different groups on the benzene ring / heterocyclic ring, near-infrared xanthracene dyes can be functionalized, achieving symmetry control of the molecular skeleton and providing novel xanthracene molecular skeletons. Furthermore, it can control the fluorescence emission wavelength, fluorescence quantum yield, and reactive oxygen species generation capacity of the near-infrared xanthracene dyes. This dye exhibits high quantum yield (up to 79.5%), low dark toxicity and phototoxicity, or can effectively generate type I (…) under near-infrared laser irradiation. 1 O2) and type II (•OH, O2• - Reactive oxygen species can enable precise visualization imaging of mitochondria and achieve highly effective treatment of tumor cells by inducing ferroptosis.

[0031] (2) The product obtained by the preparation method of the above-mentioned benzene ring / heterocyclic modified near-infrared xanthracene dye provided by the present invention specifically targets mitochondria and can be used for mitochondrial fluorescence lifetime imaging to realize the visualization of mitochondrial physiological processes.

[0032] (3) The product obtained by the preparation method of the above-mentioned benzene ring / heterocyclic modified near-infrared xanthracene dye provided by the present invention specifically targets mitochondria. After near-infrared 630 nm laser irradiation, it mainly induces ferroptosis, thereby achieving effective inhibition of cancer cells.

[0033] (4) The benzene ring / heterocyclic modification regulation strategy proposed in this invention is expected to be a universal method for developing novel high-brightness near-infrared xanthracene dyes, providing a new technical path and important breakthrough for the integrated application of super-resolution imaging, fluorescence lifetime imaging and tumor synergistic therapy. Attached Figure Description

[0034] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0035] Figure 1 This describes the synthetic routes for all xanthracene dyes in Example 1 of this invention; Figure 2 PhSX in DMSO- in Embodiment 1 of the present invention d The hydrogen NMR spectrum in 6; Figure 3 This refers to the high-resolution mass spectrometry of PhSX in Example 1 of the present invention; Figure 4 The 1H NMR spectrum of MePhSX in CDCl3 in Example 1 of this invention; Figure 5 This refers to the high-resolution mass spectrometry of MePhSX in Example 1 of the present invention; Figure 6The 1H NMR spectrum of dMAPhSX in CDCl3 in Example 1 of this invention; Figure 7 This refers to the high-resolution mass spectrometry of dMAPhSX in Example 1 of the present invention; Figure 8 In Embodiment 1 of the present invention, CAPhSX is used in DMSO- d The hydrogen NMR spectrum in 6; Figure 9 This is the high-resolution mass spectrometry of CAPhSX in Example 1 of the present invention; Figure 10 In Embodiment 1 of the present invention, 2FPhSX is used in DMSO- d The hydrogen NMR spectrum in 6; Figure 11 This refers to the high-resolution mass spectrometry of 2FPhSX in Example 1 of this invention; Figure 12 In Embodiment 1 of the present invention, 5FPhSX is used in DMSO- d The hydrogen NMR spectrum in 6; Figure 13 This refers to the high-resolution mass spectrometry of 5FPhSX in Example 1 of the present invention; Figure 14 In Embodiment 1 of the present invention o - PyAX 1H NMR spectrum of CDCl3; Figure 15 In Embodiment 1 of the present invention o - High-resolution mass spectrometry of PyAX; Figure 16 In Embodiment 1 of the present invention m - PyAX 1H NMR spectrum of CDCl3; Figure 17 In Embodiment 1 of the present invention m - High-resolution mass spectrometry of PyAX; Figure 18 In Embodiment 1 of the present invention p -PyAX 1H NMR spectrum in CD2Cl2; Figure 19 In Embodiment 1 of the present invention p - High-resolution mass spectrometry of PyAX; Figure 20 In Embodiment 1 of the present invention, IZAX is used in DMSO- d The hydrogen NMR spectrum in 6; Figure 21 This refers to the high-resolution mass spectrometry of IZAX in Example 1 of the present invention; Figure 22 The hydrogen NMR spectrum of OZAX in CDCl3 in Example 1 of this invention; Figure 23This refers to the high-resolution mass spectrometry of OZAX in Example 1 of the present invention; Figure 24 The TZAX NMR spectrum in CD3CN in Example 1 of this invention is shown. Figure 25 This refers to the high-resolution mass spectrometry of TZAX in Example 1 of the present invention; Figure 26 In Embodiment 1 of this invention, MePhSX, dMAPhSX, CAPhSX, and 2FPhSX are... o Crystal structures of PyAX, IZAX, OZAX and TZAX; Figure 27 PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX, and others are used in Embodiment 1 of this invention. o -PyAX, m -PyAX, p (A) Normalized absorption spectra of PyAX, IZAX, OZAX and TZAX in dichloromethane; (B) Normalized fluorescence spectra; (C) Fluorescence quantum yields in dichloromethane, ethanol, N,N-dimethyl sulfoxide and PBS (containing 1% N,N-dimethyl sulfoxide); Figure 28 In Embodiment 1 of the present invention, (A) under 630 nm laser irradiation, DCFH (25 μM) was used in PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX, o -PyAX, m -PyAX, p (a) Relative fluorescence intensity (I / I0-1) at different time points in the presence of PyAX, IZAX, OZAX and TZAX (5 μM); (b) Relative absorbance (A / A0) of ABDA (100 μM) at different time points in the presence of dMAPhSX and 2FPhSX (5 μM) under 630 nm laser irradiation; (c) Relative fluorescence intensity (I / I0-1) of DHE (50 μM) at different time points in the presence of dMAPhSX and 2FPhSX (5 μM) under 630 nm laser irradiation; (d) Relative fluorescence intensity (I / I0-1) of APF (20 μM) at different time points in the presence of dMAPhSX and 2FPhSX (5 μM) under 630 nm laser irradiation; 630 nm laser intensity: 30 mW / cm²; Figure 29The properties of reactive oxygen species generated in Example 1 of this invention are as follows: (A) DCFH alone (25 μM), (B) PhSX, (C) MePhSX, (D) dMAPhSX, (E) CAPhSX, (F) 2FPhSX, (G) 5FPhSX, (H) o -PyAX, (I) m -PyAX, (J) p -PyAX, (K)IZAX, (L)OZAX and (M)TZAX (5 μM) under 630 nm laser irradiation (30 mW / cm) 2 Fluorescence spectra at different time points under the influence of ) Figure 30 To generate singlet oxygen in Example 1 of the present invention ( 1 Properties of O2); (A) ABDA alone (100 μM), (B) dMAPhSX and (C) 2FPhSX under 630 nm laser irradiation (30 mW / cm²). 2 Absorption spectra at different time points under the influence of ) Figure 31 To generate commercial superoxide anions (O2•) in Example 1 of this invention - Properties of (A) standalone APF (20 μM), (B) dMAPhSX and (C) 2FPhSX under 630 nm laser irradiation (30 mW / cm²). 2 Fluorescence spectra at different time points under the influence of ) Figure 32 The properties of commercially available hydroxyl radicals (•OH) generated in Example 1 of this invention; (A) APF alone (20 μM), (B) dMAPhSX and (C) 2FPhSX under 630 nm laser irradiation (30 mW / cm²). 2 Fluorescence spectra at different time points under the influence of ) Figure 33 This is a confocal fluorescence imaging overlay image of PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX, o-PyAX, m-PyAX, p-PyAX, IZAX, OZAX, and TZAX in Example 1 of this invention with the commercial nuclear probe Hoechst 33342. Concentrations: Probes synthesized in this invention (2 μM, of which 5FPhSX concentration is 6 μM), Hoechst 33342 (1 μg / mL).

[0036] Figure 34 This is a fluorescence co-staining image of near-infrared dye (2 μM) and commercial mitochondrial green fluorescent probe MTG (200 nM) in HeLa tumor cells in Example 1 of this invention; scale bar: 10 μm; Figure 35 Different concentrations of PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, and 5FPhSX were used in Example 1 of this invention. o -PyAX, m -PyAX, p - Cell survival rate of PyAX, IZAX, OZAX and TZAX after 24 hours of incubation in HeLa tumor cells; Figure 36 This is a fluorescence lifetime imaging image of IZAX (2 μM) in HeLa tumor cells at different time points under 630 nm laser irradiation in Example 1 of the present invention; Figure 37 The following are the fluorescence lifetimes of IZAX (2 μM) in HeLa tumor cells under 630 nm laser irradiation: (A) average fluorescence lifetimes of mitochondria and nuclei at different time points; (B) fluorescence lifetime decay at 30 s in mitochondria and nuclei. Figure 38 In Example 1 of this invention, different concentrations of dMAPhSX and 2FPhAX were used in HeLa tumor cells. (A) No light exposure; (B) After 4 minutes of 630 nm light exposure (power 80 mW / cm²). 2 Cell survival rate after 24 hours of incubation; Figure 39 This is a confocal laser scanning microscopy image of reactive oxygen species (ROS) generated in HeLa tumor cells in Example 1 of this invention; HepG2 cells were stained with dMAPhSX and 2FPhAX (2 μM) and the commercial ROS probe DCFH-DA (10 μM), respectively, and then irradiated with a 630 nm laser (50 mW / cm²). 2 Confocal laser scanning microscopy images of reactive oxygen species generated under conditions of 4 minutes or in the dark.

[0037] Figure 40 This invention demonstrates the anticancer activity of dMAPhSX and 2FPhAX in Example 1. In the presence of different inhibitors, (A) dMAPhSX and (B) 2FPhAX (2.5 μM) were used in HeLa tumor cells with and without 630 nm laser irradiation (80 mW / cm²). 2 Cell survival rate after incubation for 24 hours (4 minutes). Detailed Implementation

[0038] It should be noted that the following detailed description is illustrative and intended to provide further explanation of the invention. Unless otherwise specified, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0039] To enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments and comparative examples.

[0040] Example 1: Near-infrared xanthracene dyes PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX o -PyAX, m -PyAX, p - Synthesis of PyAX, IZAX, OZAX and TZAX In this embodiment, substituents R1 and R2 are both methyl groups, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is an N,N-dimethyl group or a carboxyl group or a fluorine atom or a hydrogen atom, R9 is a methyl group or a fluorine atom or a hydrogen atom, and R... 10 R 11 R 12 All are hydrogen atoms, X is ethylene, X2 is a carbon-carbon double bond (C=C) or a nitrogen-methyl group (N-CH3) or an oxygen atom or a sulfur atom, Y1, Y2, and Y3 are all carbon atoms or nitrogen atoms, Z - It is perchlorate. The corresponding near-infrared xanthracene dyes (named PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX, respectively) o -PyAX, m -PyAX, p The chemical structures of PyAX, IZAX, OZAX, and TZAX are as follows: ; The synthetic routes for these near-infrared xanthracene dyes are as follows: Figure 1 As shown, they were synthesized via a condensation reaction of compounds 1 and 2. Their structures were confirmed by proton nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry. Figure 2-25 Additionally, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, o -PyAX, IZAX, OZAX, and TZAX have also had their structures confirmed by single crystal analysis (CCDC numbers: 2472031-2472038; perchlorate ions have been omitted for clarity of structure representation). Figure 26 ).

[0041] Synthesis of compound PhSX: Compound 1 (756 mg, 4 mmol) and benzaldehyde (212 mg, 2 mmol) were added sequentially to a round-bottom flask, followed by the addition of HClO4 (10 mL) and stirring until dissolved. The mixture was heated to 100 °C and reacted for 16 hours, then cooled to room temperature. The mixture was poured into ice water, filtered under reduced pressure to obtain a solid, and washed with cold water. The crude solid was dried in a vacuum drying oven and then purified by column chromatography using dichloromethane and ethanol (100:1 to 50:1 and 20:1) as eluents to obtain a dark green solid PhSX (621 mg, 56.7%). 1 H NMR (400 MHz, DMSO- d 6) δ (ppm): 8.06 (d, J = 9.0 Hz, 2H), 7.67–7.54 (m, 3H), 7.40 (dd, J 1 = 8.0 Hz J 2 = 1.2Hz, 2H), 6.87 (dd, J 1 = 9.0 Hz J 2 = 2.4 Hz, 2H), 6.72 (d, J = 2.0 Hz, 2H), 3.14 (s,12H), 2.88 (t, J = 7.5 Hz, 4H), 2.60 (t, J = 7.5 Hz, 4H). HRMS-ESI m / z: calcd for[C 31 H 31 N2O + ] 447.2431, found 447.2438 ([M] + ). Synthesis of compound MePhSX: Using a similar synthetic scheme as compound PhSX, compound 1 was reacted with o-methylbenzaldehyde to give a dark green solid MePhSX (588 mg, 52.4%). 1 H NMR (400 MHz, CDCl3), δ (ppm): 8.03 (d, J = 9.0 Hz, 2H), 7.46–7.34 (m, 3H), 7.09 (d, J = 7.6 Hz, 1H), 6.83 (d, J= 9.0Hz, 2H), 6.54 (s, 2H), 3.18 (s, 12H), 2.98–2.82 (m, 4H), 2.64–2.53 (m, 2H), 2.52–2.41 (m, 2H), 2.16 (s, 3H). HRMS-ESI m / z: calcd for [C 32 H 33 N2O + ] 461.2587, found 461.2577 ([M] + ). Synthesis of compound dMAPhSX: Using a similar synthetic scheme as compound PhSX, compound 1 was reacted with p-dimethylaminobenzaldehyde to give a dark green solid dMAPhSX (616 mg, 52.2%). 1 H NMR (400 MHz, CDCl3), δ (ppm): 7.95 (d, J = 9.0 Hz, 2H), 7.13 (d, J = 8.1 Hz, 2H), 6.84 (d, J = 7.2 Hz, 2H), 6.78 (d, J = 9.0 Hz, 2H), 6.53 (s, 2H), 3.16 (s, 12H), 3.08 (s, 6H), 2.93–2.85 (m, 4H), 2.85–2.77 (m, 4H). HRMS-ESI m / z: calcd for [C 33 H 36 N3O + ] 490.2853, found 490.2845 ([M] + ). Synthesis of compound CAPhSX: Using a similar synthetic scheme as compound PhSX, compound 1 was reacted with p-aldehyde benzoic acid to give a dark green solid CAPhSX (595 mg, 50.3%). 1 H NMR (400 MHz, DMSO- d 6) δ (ppm): 8.15 (d, J = 8.3 Hz, 2H), 8.07 (d, J = 9.0 Hz, 2H), 7.54 (d, J= 8.3 Hz, 2H), 6.87 (dd, J 1 = 9.1 Hz J 2 = 2.5 Hz, 2H), 6.72 (d, J = 2.3 Hz, 2H), 3.14 (s, 12H), 2.87 (t, J =7.5 Hz, 4H), 2.57 (t, J = 7.5 Hz, 4H). HRMS-ESI m / z: calcd for [C 32 H 31 N2O3 + ]491.2329, found 491.2311 ([M] + ). Synthesis of compound 2FPhSX: Using a similar synthetic scheme as compound PhSX, compound 1 was reacted with 2,6-difluorobenzaldehyde to give a dark green solid 2FPhSX (587 mg, 50.3%). 1 H NMR (400 MHz, DMSO- d 6) δ (ppm): 8.06 (d, J = 9.0 Hz, 2H), 7.81–7.71 (m, 1H), 7.43 (t, J = 8.3 Hz, 2H), 6.88 (d, J =9.1 Hz, 2H), 6.74 (s, 2H), 3.15 (s, 12H), 2.91 (t, J = 7.4 Hz, 4H), 2.61 (t, J =7.3 Hz, 4H). HRMS-ESI m / z: calcd for [C 31 H 29 F2N2O + ] 483.2242, found 483.2236([M] + ). Synthesis of compound 5FPhSX: Using a similar synthetic scheme as compound PhSX, compound 1 was reacted with 2,3,4,5,6-tetrafluorobenzaldehyde to give a dark green solid 5FPhSX (536 mg, 42.1%). 1 H NMR (400 MHz, DMSO- d 6) δ(ppm): 8.06 (d, J = 9.1 Hz, 2H), 6.89 (dd, J 1 = 9.1 Hz J 2 = 2.5 Hz, 2H), 6.76 (d, J =2.3 Hz, 2H), 3.16 (s, 12H), 2.91 (t, J = 7.4 Hz, 4H), 2.69 (t, J = 7.4 Hz, 4H). HRMS-ESI m / z: calcd for [C 31 H 26 F5N2O + ] 537.1960, found 537.1958 ([M] + ). compound o Synthesis of -PyAX: Using a similar synthetic scheme to that of compound PhSX, compound 1 reacted with 2-aldehyde pyridine to give a dark green solid. o -PyAX (216 mg, 19.7%). 1 H NMR (400 MHz, CDCl3), δ (ppm): 8.77(dt, J 1 = 4.7 Hz J 2= ​​1.2 Hz, 1H), 8.10–8.03 (m, 2H), 7.88 (d, J = 9.0 Hz, 1H), 7.66 (d, J = 9.1 Hz, 1H), 7.54–7.47 (m, 1H), 6.79–6.70 (m, 2H), 6.59 (dd, J 1 = 14.3 Hz J 2 = 2.5 Hz, 2H), 3.34 (t, J = 7.3 Hz, 2H), 3.22 (t, J = 6.8 Hz, 2H), 3.19(s, 6H), 3.17 (s, 6H), 3.01 (t, J = 7.2 Hz, 2H), 2.83 (t, J = 6.7 Hz, 2H). HRMS-ESI m / z: calcd for [C 30 H 30N3O + ] 448.2383, found 448.2385 ([M] + ). compound m Synthesis of -PyAX: Using a similar synthetic scheme to that of compound PhSX, compound 1 reacted with 3-aldehyde pyridine to give a dark green solid. m -PyAX (240 mg, 21.9%). 1 H NMR (400 MHz, CDCl3), δ (ppm): 8.93 (s, 1H), 8.82 (d, J = 2.9 Hz, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.79 (d, J = 9.0 Hz, 1H), 7.69 (d, J = 9.1 Hz, 1H), 7.63 (dd, J 1 = 7.7 Hz J 2 = 4.6 Hz, 1H), 6.76 (dd, J 1 = 9.1 Hz J 2 = 2.5 Hz, 1H), 6.68 (dd, J 1 = 9.0 Hz J 2 = 2.4 Hz, 1H), 6.60 (dd, J 1 = 8.2Hz J 2 = 2.2 Hz, 2H), 3.37 (t, J = 7.2 Hz, 2H), 3.18 (s, 12H), 3.04 (t, J = 7.2 Hz,2H), 2.98–2.90 (m, 2H), 2.90–2.83 (m, 2H). HRMS-ESI m / z: calcd for [C 30 H 30 N3O + ]448.2383, found 448.2389 ([M] + ). compound p Synthesis of -PyAX: Using a similar synthetic scheme to that of compound PhSX, compound 1 reacted with 4-aldehyde pyridine to give a dark green solid. p -PyAX (236 mg, 21.5%). 1H NMR (400 MHz, CD2Cl2), δ (ppm): 8.89 (d, J = 4.2 Hz, 2H), 7.84 (d, J = 9.0 Hz, 1H), 7.78–7.61 (m, 3H), 6.86–6.72 (m,2H), 6.64 (d, J = 17.8 Hz, 2H), 3.36 (t, J = 7.1 Hz, 2H), 3.19 (s, 12H), 3.01 (t, J = 7.1 Hz, 2H), 2.98–2.90 (m, 2H), 2.90–2.78 (m, 2H). HRMS-ESI m / z: calcd for[C 30 H 30 N3O + ] 448.2383, found 448.2390 ([M] + ). Synthesis of compound IZAX: Using a similar synthetic scheme as that of compound PhSX, compound 1 was reacted with 1-methyl-1H-imidazol-2-carboxaldehyde to give dark green solid IZAX (245 mg, 22.2%). 1 H NMR (400 MHz, DMSO- d 6) δ (ppm): 7.92 (d, J = 9.0 Hz, 1H), 7.74 (d, J = 9.7 Hz, 1H), 7.62 (d, J = 0.8 Hz, 1H), 7.29 (d, J = 1.0 Hz, 1H), 6.84–6.78 (m, 3H), 6.76 (d, J = 2.3 Hz, 1H), 3.99(s, 3H), 3.29 (t, J = 7.2 Hz, 2H), 3.15 (s, 6H), 3.14 (s, 6H), 3.05 (t, J = 6.7Hz, 2H), 2.94 (t, J = 7.2 Hz, 2H), 2.80 (t, J = 6.6 Hz, 2H). HRMS-ESI m / z: calcdfor [C 29 H 31 N4O + ] 451.2492, found 451.2492 ([M] + ). Synthesis of compound OZAX: Using a similar synthetic scheme as that of compound PhSX, compound 1 was reacted with oxazol-2-carboxaldehyde to give a dark green solid OZAX (198 mg, 18.4%). 1 H NMR (400 MHz, CDCl3), δ (ppm): 8.05 (s, 1H), 7.97 (d, J = 9.0 Hz, 1H), 7.71 (d, J = 9.2 Hz, 1H), 7.49 (s, 1H), 6.80 (dd, J 1 = 9.2 Hz J 2 = 2.7 Hz, 1H), 6.73 (dd, J 1 = 9.1 Hz J 2 = 2.5 Hz, 1H), 6.61 (dd, J 1 = 12.7 Hz J 2= ​​2.5 Hz, 2H), 3.41–3.33 (m, 4H), 3.21 (s, 6H), 3.20 (s, 6H), 3.02(t, J = 7.3 Hz, 2H), 2.90 (t, J = 6.8 Hz, 2H). HRMS-ESI m / z: calcd for [C 28 H 28 N3O2 + ]438.2176, found 438.2166 ([M] + ). Synthesis of compound TZAX: Using a similar synthetic scheme as that of compound PhSX, compound 1 was reacted with thiazole-2-carboxaldehyde to give a dark green solid TZAX (214 mg, 19.3%). 1 H NMR (400 MHz, CD3CN), δ (ppm): 8.14 (d, J = 2.2 Hz, 1H), 7.94 (d, J = 2.1 Hz, 1H), 7.90 (d, J= 9.0 Hz, 1H), 7.67 (d, J = 9.7Hz, 1H), 6.84 (d, J = 9.0 Hz, 1H), 6.78–6.72 (m, 2H), 6.68 (s, 1H), 3.42 (t, J =6.7 Hz, 2H), 3.28 (t, J = 7.3 Hz, 2H), 3.15 (s, 12H), 2.94 (t, J = 7.2 Hz, 2H), 2.86 (t, J = 6.7 Hz, 2H). HRMS-ESI m / z: calcd for [C 28 H 28 N3OS + ] 454.1948, found454.1944 ([M] + ). Near-infrared anthracene dyes PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX o -PyAX, m - PyAX, p Photophysical properties of PyAX, IZAX, OZAX and TZAX PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, and 5FPhSX from Example 1 were used. o -PyAX, m -PyAX, p PyAX, IZAX, OZAX, and TZAX were added sequentially to dichloromethane solvent, and ultraviolet absorption spectra were measured. Figure 27 As shown in AB, from PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX, o -PyAX, m -PyAX, p The absorption spectra of PyAX, IZAX, OZAX, and TZAX show that their maximum absorption wavelengths in dichloromethane are between 595 and 682 nm, and the absorption peaks extend into the near-infrared region (650-700 nm), which is very advantageous for excitation or phototherapy using near-infrared lasers.

[0042] Testing PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, and 5FPhSX using the integrating sphere method o -PyAX, m -PyAX, p -Absolute quantum yields of PyAX, IZAX, OZAX, and TZAX in solvents of different polarities. For example... Figure 27As shown in C, different substituents have a significant impact on the fluorescence quantum yield of the dye. Among them, MePhAX exhibits a fluorescence quantum yield as high as 79.5%, which has the potential to be applied to STED super-resolution microscopy, which has high requirements for fluorescence quantum yield.

[0043] Near-infrared anthracene dyes PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX o -PyAX, m - PyAX, p Study on the Reactive Oxygen Generation Properties of PyAX, IZAX, OZAX and TZAX under Near-Infrared Laser Excitation The reactive oxygen species (ROS) generation efficiency of near-infrared xanthracene dyes was tested using a commercial ROS indicator (2',7'-dichlorodihydrofluorescein, DCFH) under 630 nm near-infrared laser irradiation. Figure 28 A and Figure 29 As shown, with the increase of 630nm near-infrared laser irradiation time, the concentrations of PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, and 5FPhSX... o -PyAX, m -PyAX, p The fluorescence intensity of DCFH in PBS solutions of PyAX, IZAX, OZAX, and TZAX dyes showed varying degrees of enhancement from 0 to 240 seconds. This indicates that they possess different abilities to generate reactive oxygen species (ROS). Specifically, dMAPhSX showed a 366-fold increase in ROS generation compared to baseline DCFH, and 2FPhSX showed a 213-fold increase, demonstrating exceptionally superior ROS generation performance.

[0044] Using commercial singlet oxygen ( 1 O2 scavenger 9,10-anthratridimyl-bis(methylene)dimalonic acid (ABDA) was used to scavenge singlet oxygen (dMAPhSX) and 2FPhSX in PBS under 630 nm near-infrared laser irradiation. 1 O2 generation efficiency test experiment. For example... Figure 28 B and Figure 30 As shown, the attenuation of the characteristic absorption peak at 380 nm can be observed. 1 O2-mediated oxidative cleavage revealed different photosensitization efficiencies. The degradation rate curves of ABDA showed that both dMAPhSX and 2FPhSX exhibited strong [efficiencies / activities]. 1 O2 generation capability.

[0045] Using commercial superoxide anions (O2• - The scavenger 9, dihydroethidium (DHE), under 630 nm near-infrared laser irradiation, carried out the scavenging of superoxide anions (O2•) in PBS by dMAPhSX and 2FPhSX. - Generation efficiency test experiment. (For example...) Figure 28 C and Figure 31As shown, with increasing 630 nm near-infrared laser irradiation time, the fluorescence intensity of DHE in PBS solutions containing dMAPhSX and 2FPhSX increased to varying degrees from 0 to 240 seconds at 600 nm. This indicates that they generate O2• to different degrees. - The ability.

[0046] The •OH generation efficiency of dMAPhSX and 2FPhSX was tested using the commercial hydroxyl radical (•OH) probe 3'-p-aminophenylfluorescein (APF) under 630 nm near-infrared laser irradiation. Figure 28 D and Figure 32 As shown, under 630 nm near-infrared laser irradiation, the fluorescence intensity of APF solutions containing dMAPhSX and 2FPhSX increased rapidly, exhibiting a strong •OH generation ability. In summary, in xanthracene... meso Introducing different substituents at different positions on the benzene ring can modulate the reactive oxygen species (ROS) generation capacity, simultaneously activating both type I and type II photodynamic activities. Among them, dMAPhSX and 2FPhSX exhibit superior ROS generation capacity compared to other compounds, indicating that regioisomer engineering plays a crucial role in the fine-tuning of triplet excited state energy levels.

[0047] Near-infrared anthracene dyes PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, 5FPhSX o -PyAX, m - PyAX, p Confocal Imaging Study of PyAX, IZAX, OZAX and TZAX in HeLa Tumor Cells HeLa cells were seeded into confocal microplates and incubated at 37 °C with 5% CO2 for 24 hours. Then, the old culture medium was aspirated from the confocal microplates, and cells were incubated with 2 μM near-infrared xanthracene dyes PhSX, MePhSX, dMAPhSX, CAPhSX, 2FPhSX, and 5FPhSX. o -PyAX, m -PyAX, p HeLa cells were incubated with PyAX, IZAX, OZAX, and TZAX in a medium containing 1 μg / mL of the commercial nuclear staining agent Hoechst 33342 for 30 min before imaging. Figure 33 As shown, experimental results indicate that after PhSX, MePhSX, dMAPhSX, 2FPhSX, 5FPhSX, o -PyAX, m -PyAX, p PyAX, IZAX, OZAX, and TZAX showed bright fluorescence in the perinuclear cytoplasm stained with commercial Hoechst 33342, indicating their high cell penetration and excellent counterstaining ability. In contrast, CAPhSX showed no fluorescence in the perinuclear cytoplasm stained with commercial Hoechst 33342, indicating its inability to enter the cell.

[0048] Due to the negative membrane potential of mitochondria, cationic dyes typically tend to target mitochondria. Using HeLa tumor cells as an example, we validated the effectiveness of near-infrared xanthracene dyes PhSX, MePhSX, dMAPhSX, 2FPhSX, and 5FPhSX through co-staining experiments with the commercially available mitochondrial green fluorescent probe MTG. o -PyAX, m -PyAX, p -Targeting of PyAX, IZAX, OZAX, and TZAX in mitochondria. HeLa cells were seeded into confocal microplates and incubated at 37 °C and 5% CO2 for 24 h. Then, the old culture medium was aspirated from the confocal microplates, and the cells were incubated with 200 nM MTG at 37 °C and 5% CO2 for 1 h. Subsequently, the cells were incubated with 2 μM PhSX, MePhSX, dMAPhSX, 2FPhSX, and 5FPhSX (6 μM). o -PyAX, m -PyAX, p Incubate with PyAX, IZAX, OZAX, and TZAX together for 15 minutes. Aspirate the old culture medium from the confocal dish and wash three times with PBS. Imaging is performed using confocal laser scanning microscopy. Figure 34 As shown, the red fluorescence of near-infrared xanthracene dyes clearly outlines the round and elongated mitochondrial contours and shows significant overlap with the green fluorescence of MTG, further confirmed by their high Pearson correlation coefficients (0.78–0.87). These results indicate that these near-infrared xanthracene dyes possess excellent mitochondrial targeting capabilities in cancer cells.

[0049] Near-infrared anthracene dyes PhSX, MePhSX, dMAPhSX, 2FPhSX, 5FPhSX o -PyAX, m -PyAX, p - Dark toxicity studies of PyAX, IZAX, OZAX and TZAX in HeLa tumor cells In HeLa cells, different concentrations of near-infrared xanthracene dyes PhSX, MePhSX, dMAPhSX, 2FPhSX, and 5FPhSX were used. o -PyAX, m -PyAX, p - Dark cytotoxicity assays using PyAX, IZAX, OZAX, and TZAX. HeLa cells were seeded into 96-well plates and incubated at 37 °C with 5% CO2 for 24 hours. Then, the old culture medium was aspirated from the 96-well plates, and different concentrations of PhSX, MePhSX, dMAPhSX, 2FPhSX, and 5FPhSX were added. o -PyAX, m -PyAX, pCells were cultured in PyAX, IZAX, OZAX, and TZAX media. After 24 hours, the old medium was aspirated from the 96-well plate, and medium containing 20 μL of MTT was added. The cells were incubated at 37 °C and 5% CO2 for 4 hours. Then, the old medium was aspirated, and 100 μL of DMSO was added. Cell viability was assessed using a microplate reader. Figure 35 As shown, PhSX, MePhSX, and dMAPhSX exhibited high cell viability with negligible toxicity. However, 2FPhSX and 5FPhSX... o -PyAX, m -PyAX, p -PyAX, IZAX, OZAX, and TZAX all exhibited varying degrees of concentration-dependent cytotoxicity. This suggests that MePhSX has extremely high potential for bioimaging applications; while dMAPhSX, 2FPhSX, and p -PyAX has the potential for application in photodynamic therapy and / or chemotherapy for cancer treatment.

[0050] Lifetime Imaging Study of Near-Infrared Xanthracene Dye IZAX in HeLa Tumor Cells Based on the significant organelle-level migration of the near-infrared xanthracene dye IZAX under light illumination, we conducted a lifetime imaging study of IZAX in HeLa tumor cells under light illumination. HeLa cells were seeded into confocal microplates and incubated at 37 °C and 5% CO2 for 24 hours. Then, the old culture medium was aspirated from the confocal microplates, and the HeLa cells were incubated with 2 μM IZAX for 30 min before lifetime imaging. Figures 36-37 As shown, with continuous irradiation by a 630 nm near-infrared laser, the fluorescence lifetime of mitochondria gradually increased from 0.75 ns to 1.33 ns; while the cell nucleus gradually exhibited fluorescence, and its fluorescence lifetime also gradually increased from 1.34 ns to 1.87 ns. This is because IZAX induces changes in the mitochondrial membrane potential under illumination, thereby causing IZAX to migrate towards the cell nucleus. This indicates that the near-infrared xanthracene dye IZAX has high application potential in the study of mitochondrial membrane potential.

[0051] Phototoxicity study of near-infrared xanthane dyes dMAPhSX and 2FPhSX in HeLa tumor cells Based on studies on the reactive oxygen species generation properties of near-infrared xanthracene dyes under near-infrared laser excitation and their dark toxicity in HeLa tumor cells, dMAPhSX and 2FPhSX have been found to have potential applications in photodynamic therapy and / or chemotherapy for cancer treatment. Therefore, the phototoxicity of different concentrations of dMAPhSX and 2FPhSX in HeLa tumor cells was further investigated. HeLa cells were seeded into 96-well plates and cultured at 37 °C and 5% CO2 for 24 hours. Then, the old culture medium in the 96-well plates was aspirated, and culture medium containing different concentrations of dMAPhSX and 2FPhSX was added, respectively. The cells were then irradiated with a 630 nm near-infrared laser for 4 minutes (power 80 mW / cm²). 2 After 24 hours, the old culture medium was aspirated from the 96-well plate, and medium containing 20 μL of MTT was added. The cells were incubated at 37 °C and 5% CO2 for 4 hours. Then, the old culture medium was aspirated, and 100 μL of DMSO was added. Cell viability was measured using a microplate reader. Figure 38 As shown, both dMAPhSX and 2FPhSX exhibited significant concentration-dependent cytotoxicity.

[0052] Study on the reactive oxygen species generation properties of near-infrared xanthanthracene dyes dMAPhSX and 2FPhSX in HeLa tumor cells In HeLa cells, in vitro ROS generation experiments using dMAPhSX and 2FPhSX were performed. HeLa cells were stained with 20 μM of the indicator 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) for 1 hour, followed by staining with 1 μg / mL of the commercial nuclear staining agent Hoechst 33342 and 2 μM of dMAPhSX and 2FPhSX, respectively. Cells were incubated at 37 °C and 5% CO2 for 1 hour. The cells were then stained with a 630 nm near-infrared laser (50 mW / cm²). 2 Irradiate for 4 minutes and then protect from light for 4 minutes. Aspirate the old culture medium from the confocal dish, wash three times with PBS, and image using CLSM. Figure 39 As shown, HeLa cells incubated with DCFH-DA exhibited almost no green fluorescence under dark conditions or 630 nm laser irradiation. When HeLa cells were co-incubated with dMAPhSX and 2FPhSX under dark conditions, the green fluorescence of DCFH-DA was very weak. However, after 4 minutes of 630 nm laser irradiation, HeLa cells co-incubated with dMAPhSX and 2FPhSX showed significantly enhanced green fluorescence, indicating that photo-triggered ROS generation was more significant than that induced by chemotherapy alone. This further suggests that dMAPhSX and 2FPhSX have potential applications in photodynamic therapy and / or chemotherapy for cancer treatment.

[0053] Study on the anticancer mechanism of near-infrared xanthane dyes dMAPhSX and 2FPhSX on HeLa tumor cells Cell death pathway validation experiments were performed in HeLa cells. Cells were incubated with various inhibitors, including cycloheximide (CHX, a paraapoptosis inhibitor), ferrostatin-1 (Fer-1, a ferroptosis inhibitor), z-VAD-FMK (ZVAD, an apoptosis inhibitor), necrostatin-14 (Nec-14, a necrosis inhibitor), disulfiram (a pyroptosis inhibitor), and autophinib (an autophagy inhibitor). Then, 2.5 μM dMAPhSX and 2FPhSX were added for phototoxicity testing (630 nm laser, 80 mW / cm²). 2 Experimental test (4 minutes of light exposure). Figure 40 As shown in the inhibitor experiments, the addition of the paraapoptosis inhibitor Fer-1 significantly improved cell survival in phototoxicity studies. It should be noted that the addition of the pyroptosis inhibitor disulfiram also slightly improved the survival of 2FPhSX cells. This indicates that under light irradiation, dMAPhSX and 2FPhSX primarily induce ferroptosis, thus achieving a synergistic drug-photodynamic therapy against cancer cells.

[0054] Example 2 The difference from Example 1 is that: substituents R1 and R2 are both methyl, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is N,N-dimethyl or carboxyl or fluorine or hydrogen atom, R9 is methyl or fluorine or hydrogen atom, R... 10 R 11 R 12 All are hydrogen atoms, X1 is O, X2 is a carbon-carbon double bond (C=C) or a nitrogen-methyl group (N-CH3) or an oxygen atom or a sulfur atom, Y1, Y2, and Y3 are all carbon atoms or nitrogen atoms, Z - For PF6 - The synthesis method is the same as in Example 1.

[0055]

[0056] The compound in Example 2 exhibits red-shifted fluorescence with a small change in fluorescence peak wavelength, similar to the dye in Example 1. The main difference lies in the band gap, which is as long as 750 nm. The fluorescence quantum yield of its symmetrical structure dye can reach up to 88.3%.

[0057] Example 3 The difference from Example 1 is that: substituents R1 and R2 are both methyl, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is N,N-dimethyl or carboxyl or fluorine or hydrogen atom, R9 is methyl or fluorine or hydrogen atom, R... 10 R 11 R12 All are hydrogen atoms, X1 is X2 is a carbon-carbon double bond (C=C), a nitrogen-methyl group (N-CH3), an oxygen atom, or a sulfur atom; Y1, Y2, and Y3 are all carbon or nitrogen atoms; Z - For PF6 - The synthesis method is the same as in Example 1.

[0058] .

[0059] The compounds in Example 3 exhibit stronger fluorescence with minimal variation in fluorescence peak wavelength, similar to the dyes in Example 1. The main difference lies in the band gap, which is not significantly different between them, reaching a maximum of 680 nm. The fluorescence quantum yield of the symmetric structure dyes can reach 94.6%, and the singlet oxygen yield under illumination of dyes I-15, I-17, and I-24 can reach 32.8%.

[0060] Example 4 The difference from Example 1 is that: substituents R1 and R2 are both methyl, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is N,N-dimethyl or carboxyl or fluorine or hydrogen atom, R9 is methyl or fluorine or hydrogen atom, R... 10 R 11 R 12 All are hydrogen atoms, X1 is S, X2 is a carbon-carbon double bond (C=C) or a nitrogen-methyl group (N-CH3) or an oxygen atom or a sulfur atom, Y1, Y2, and Y3 are all carbon atoms or nitrogen atoms, Z - For PF6 - The synthesis method is the same as in Example 1.

[0061] .

[0062] The compounds in Example 4 exhibit red-shifted fluorescence with minimal changes in fluorescence peak wavelength, similar to the dyes in Example 1. The main difference lies in the band gap, which is not significantly different between them, reaching a maximum of 760 nm. The fluorescence quantum yield of the symmetric structure dyes can reach 72.5%, and the singlet oxygen yield under illumination of dyes I-27, I-29, and I-36 can reach 55.6%.

[0063] Example 5 The difference from Example 1 is that: substituents R1 and R2 are both methyl, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is N,N-dimethyl or carboxyl or fluorine or hydrogen atom, R9 is methyl or fluorine or hydrogen atom, R... 10 R 11 R 12 All are hydrogen atoms, X1 is X2 is a carbon-carbon double bond (C=C), a nitrogen-methyl group (N-CH3), an oxygen atom, or a sulfur atom; Y1, Y2, and Y3 are all carbon or nitrogen atoms; Z - For PF6 - The synthesis method is the same as in Example 1.

[0064] ; The compounds in Example 5 exhibit red-shifted fluorescence with minimal changes in fluorescence peak wavelength, similar to the dyes in Example 1. The main difference lies in the band gap, which is not significantly different between them, reaching a maximum of 720 nm. The fluorescence quantum yield of the symmetric structure dyes can reach 65.2%, and the singlet oxygen yield under illumination of dyes I-39, I-41, and I-48 can reach 38.8%.

[0065] Example 6 The difference from Example 1 is that: substituents R1 and R2 are both methyl, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is N,N-dimethyl or carboxyl or fluorine or hydrogen atom, R9 is methyl or fluorine or hydrogen atom, R... 10 R 11 R 12 All are hydrogen atoms, X1 is X2 is a carbon-carbon double bond (C=C), a nitrogen-methyl group (N-CH3), an oxygen atom, or a sulfur atom; Y1, Y2, and Y3 are all carbon or nitrogen atoms; Z - For PF6 - The synthesis method is the same as in Example 1.

[0066] ; The compounds in Example 6 exhibit red-shifted fluorescence with minimal changes in fluorescence peak wavelength, similar to the dyes in Example 1. The main difference lies in the band gap, which is not significantly different between them, reaching a maximum of 738 nm. The fluorescence quantum yield of the symmetric structure dyes can reach 60.5%, and the singlet oxygen yield under illumination of dyes I-51, I-53, and I-60 can reach 40.2%.

[0067] Example 7 The difference from Example 1 is that: substituents R1 and R2 are both butyl, R3 and R4 are both hydrogen atoms, R5, R6, and R8 are fluorine atoms or hydrogen atoms, R7 is N,N-dimethyl or carboxyl or fluorine or hydrogen atom, R9 is methyl or fluorine or hydrogen atom, R... 10 R 11 R 12 All are hydrogen atoms, X1 is X2 is a carbon-carbon double bond (C=C), a nitrogen-methyl group (N-CH3), an oxygen atom, or a sulfur atom; Y1, Y2, and Y3 are all carbon or nitrogen atoms; Z - For PF6- The synthesis method is the same as in Example 1.

[0068] ; The compound in Example 7 exhibits red-shifted fluorescence with a small change in fluorescence peak wavelength, similar to the dye in Example 1. The main difference lies in the band gap, which is as long as 760 nm. The fluorescence quantum yield of its symmetrical structure dye can reach 50.5%.

[0069] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A near-infrared oxanthracene dye with tunable structural symmetry, characterized in that: Its structural formula is shown in formula Ia or Ib: ； Among them, R1, R2, R5 to R 12 It is independently selected from hydrogen, C1-C6 straight-chain hydrocarbon group, C1-C6 branched hydrocarbon group, C1-C6 alkoxy group, carboxylic acid ester group, benzene ring, and any one of C1-C6 alkyl, alkoxy, substituted thiophene and substituted benzene ring optionally substituted with C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkynyl, C1-C4 alkynyloxy, ethyl acetate group, cyano, trifluoromethyl and halogen substituted C1-C6 alkyl, alkoxy, substituted thiophene and substituted benzene ring; R3 and R4 are independently selected from any one of hydrogen, C1-C6 alkyl, fluorine, chlorine, bromine, iodine, cyano, trifluoromethyl, and C1-C6 alkoxy. X1 and X2 are independently selected , , , , , , , , , , , , one of the; Z - Independently selected from halide ions, ClO4 - PF6 - BPh4 - N(CN)2 - and BF4 - Any one of them.

2. The near-infrared oxanthracene dye with tunable structural symmetry according to claim 1, characterized in that: R1 and R2 are independently selected from hydrogen and C1-C6 straight-chain alkyl groups; Preferably, R1 and R2 are independently selected from C1-C3 straight-chain alkyl groups.

3. The method for preparing the benzene ring-modified near-infrared oxanthracene dye according to claim 1 or 2, characterized in that: Its synthesis process is as follows: ； Compound 1 and Compound 2 were added to HClO4 in a certain proportion and heated to react at a temperature of 90-110℃ for 16-20 h. After the reaction was completed, the mixture was cooled and an aqueous solution of Z salt or Z acid was added. After purification, the product was obtained. Z - Independently selected from halide ions, ClO4 - PF6 - BPh4 - N(CN)2 - and BF4 - Any one of them.

4. The method for preparing benzene ring-modified near-infrared xanthracene dyes according to claim 3, characterized in that: The molar ratio of compound 1 to compound 2 is 1.5-2.5:1; Preferably, the molar ratio of compound 1 to compound 2 is 1.8-2.3:1; Preferably, the molar ratio of compound 1 to compound 2 is 2:

1.

5. The method for preparing benzene ring-modified near-infrared xanthracene dyes according to claim 3, characterized in that: The reaction temperature of compound 1 and compound 2 is 95-110℃, preferably 100-110℃, and more preferably 100-105℃; Alternatively, the concentration of compound 2 in HClO4 is 0.1-1 mol / L; Alternatively, the purification method may be a silica gel column separation and purification method; Preferably, the eluent used for silica gel column separation and purification is a mixture of dichloromethane and ethanol, with a volume ratio of dichloromethane to ethanol of 20-100:

1.

6. The method for preparing benzene ring-modified near-infrared oxanthracene dyes according to claim 3, characterized in that: The molar ratio of Z salt or Z acid to compound 2 is 3-6:

1.

7. The method for preparing the heterocyclic-modified near-infrared oxanthracene dye according to claim 1 or 2, characterized in that: Its synthesis process is as follows: ； Compound 1 and compound 3 were added to HClO4 in a certain proportion and heated to react at a temperature of 90-110 °C for 16-20 h. After the reaction was completed, the mixture was cooled and an aqueous solution of Z salt or Z acid was added. After purification, the product was obtained.

8. The method for preparing heterocyclic-modified near-infrared oxanthracene dyes according to claim 7, characterized in that: The molar ratio of compound 1 to compound 3 is 1.5-2.5:1; Preferably, the molar ratio of compound 1 to compound 3 is 1.8-2.3:1; Preferably, the molar ratio of compound 1 to compound 3 is 2:

1.

9. The method for preparing heterocyclic-modified near-infrared oxanthracene dyes according to claim 7, characterized in that: The concentration of compound 3 in HClO4 is 0.1-1 mol / L; Preferably, the purification method is a silica gel column separation and purification method; Preferably, the eluent used for silica gel column separation and purification is a mixture of dichloromethane and ethanol, with a volume ratio of dichloromethane to ethanol of 20-100:

1. Alternatively, the molar ratio of Z salt or Z acid to compound 3 is 3-6:

1.

10. The application of the benzene ring / heterocyclic modified near-infrared xanthracene dyes of claims 1-2, or the benzene ring / heterocyclic modified near-infrared xanthracene dyes prepared by any of the preparation methods of claims 3-9, in mitochondrial imaging, preparation of anticancer drugs, and photodynamic therapy.

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