A type of near-infrared second-zone fluorescent dye and its preparation method and anti-tumor application

Abstract

A class of near-infrared (NIR) II fluorescent dyes, their preparation methods, and their anti-tumor applications belong to the field of fine chemical technology. These dyes have a primary absorption band above 800 nanometers, with fluorescence emission peaks covering both NIR I and II, and exhibit a large Stokes shift (up to 160 nanometers in DMF). These dyes can be used for NIR II optical imaging, avoiding defects such as dye self-absorption, autofluorescence, and fluorescence scattering. When excited by NIR light (808 nm), these dyes generate sufficient reactive oxygen species and heat, enabling them to be used in NIR II fluorescence-guided optical therapy. Furthermore, these dyes can target the specific phospholipid cardiolipin in the inner mitochondrial membrane of cells, interfering with the cardiolipin domain, affecting the activity of cardiolipin-binding proteins, and promoting cell apoptosis. These dyes can be used as novel anti-tumor chemotherapy drugs for cancer treatment.

Description

Technical Field

[0001] The present invention relates to a near-infrared second-zone fluorescent dye, a preparation method and anti-tumor application thereof, and belongs to the technical field of fine chemical industry. Background Art

[0002] Photodynamic therapy and photothermal therapy are emerging tumor treatment technologies that use light and reactive oxygen species and heat generated by photosensitizers to kill tumor cells. Traditional photodynamic therapy uses short-wavelength photosensitizers with low penetration depth and poor therapeutic effects. Photothermal conversion agents also have problems such as poor photostability and low photothermal conversion efficiency. In order to improve the therapeutic effect, researchers combine phototherapy with chemotherapy and use the fluorescence properties or photoacoustic signals of photosensitizers to track drugs in order to achieve the desired therapeutic effect. However, the method of compounding photosensitizers with chemotherapy drugs is relatively complicated and has poor reproducibility, which is not conducive to future clinical transformation. Therefore, it is necessary to develop a drug that can achieve phototherapy effects while also exerting chemotherapy effects, avoiding the various defects of nanomedicines.

[0003] At the same time, the fluorescent properties of the photosensitizer itself can be used to achieve real-time tracking of drugs and find the best treatment node. The absorption and emission wavelengths of traditional photosensitizers are both in the visible region. Since cells have strong absorption and scattering capabilities for visible light, the collected fluorescence signals will be weakened to a certain extent; the presence of strong autofluorescence in cells will also cause great interference to detection and imaging to a certain extent; and the penetration ability of visible light is limited, and signals will be lost during in vivo imaging, which is very unfavorable for drug tracking and visualization. The near-infrared second-zone optical imaging (1000-1700nm) developed in recent years has a deeper penetration ability of up to 1cm, and biological tissues have poor light absorption and scattering capabilities in this area, which can achieve high signal-to-noise ratio imaging. Therefore, it is necessary to develop a class of near-infrared second-zone fluorescent dyes that have both chemotherapy and phototherapy effects to overcome the various defects faced in imaging and treatment. Summary of the Invention

[0004] The purpose of the present invention is to prepare a class of fluorescent compounds that can be used for near-infrared region II fluorescence guidance. The main absorption of this type of dye is above 800 nanometers, and the fluorescence emission peak covers near-infrared region I and region II, with a large Stokes shift (up to 160 nanometers in DMF). This type of dye can be used for near-infrared region II optical imaging, avoiding defects such as self-absorption of the dye, spontaneous fluorescence and fluorescence scattering of the organism. This type of dye can generate sufficient reactive oxygen and heat under the excitation of near-infrared light (808nm), and can be used for optical therapy guided by near-infrared region II fluorescence. At the same time, this type of dye can target the cell mitochondrial inner membrane-specific phospholipid-cardiolipin through its amphiphilic structure, interfere with the cardiolipin domain, affect the activity of cardiolipin binding protein, promote cell apoptosis, and can be used as a new type of anti-tumor chemotherapy drug for cancer treatment.

[0005] The dyes of the present invention can avoid a series of defects faced by commercial dyes due to their excellent spectral properties, overcome the problems of traditional commercial dyes such as short absorption wavelength and high cell phototoxicity, and have wide applications in biological imaging, chemotherapy and phototherapy.

[0006] The present invention provides a compound or a pharmaceutically acceptable salt of Formula I:

[0007]

[0008] wherein n is independently 1, 2 or 3.

[0009] R1 is independent of each other Or N, when R1 is N, it connects with the carbon atoms on the left and right adjacent positions of the carbon atom on the substituted benzene ring to form a six-membered ring structure.

[0010] When R1 in Formula I is N, the compound structures of the six-membered ring structures formed by connecting with the carbon atoms on the left and right adjacent positions of the substituted benzene ring are as follows:

[0011] R2 is or

[0012] wherein p is an integer of 1-5, t is an integer of 1-3 (R3 replaces at least one position on the benzene ring, and R4 replaces at least one position on the five-membered heterocyclic ring), and R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C20 alkoxy, C1-C20 alkyl, C1-C20 carboxylalkyl, C1-C20 aminoalkyl, C1-C20 hydroxyalkyl, or

[0013] R5 is a carboxyl-substituted alkoxy group having 1-10 carbon atoms, a carboxyl-substituted alkylamino group having 1-10 carbon atoms, or an amino-substituted alkylamino group having 1-10 carbon atoms; R6 is hydrogen, carboxyl, or a C1-C20 alkyl group. Z is independently O, S, or NR8.

[0014] R7 and R8 are each independently hydrogen, an alkyl group having 1 to 18 carbon atoms, a carboxyalkyl group having 1 to 18 carbon atoms, an aminoalkyl group having 1 to 18 carbon atoms, or a hydroxyalkyl group having 1 to 18 carbon atoms.

[0015] m and q are each independently an integer from 0 to 18.

[0016] X -and Y - is an anion selected from PF 6- 、BF4 - 、Cl - Br - , I - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - CF3SO3 - .

[0017] described The total positive charge is equal to X - The total amount of negative charge; The total positive charge is equal to Y - The total amount of negative charge. X - and Y - They may be the same anion or different anions.

[0018] For some optional structures of the compound of formula I, R2 is or

[0019] Wherein, p is an integer of 1-2, t is an integer of 1-2 (R3 replaces 1-2 hydrogen atoms on the benzene ring, and R4 replaces 1-2 hydrogen atoms on the five-membered heterocyclic ring), R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C20 alkoxy, C1-C20 alkyl, C1-C20 carboxylalkyl, C1-C20 aminoalkyl, C1-C20 hydroxyalkyl, or

[0020] R5 is a carboxyl-substituted alkoxy group having 1-10 carbon atoms, a carboxyl-substituted alkylamino group having 1-10 carbon atoms, or an amino-substituted alkylamino group having 1-10 carbon atoms; R6 is hydrogen, carboxyl, or a C1-C20 alkyl group. Z is independently O, S, or NR8.

[0021] R7 and R8 are each independently hydrogen, an alkyl group having 1 to 18 carbon atoms, a carboxyalkyl group having 1 to 18 carbon atoms, an aminoalkyl group having 1 to 18 carbon atoms, or a hydroxyalkyl group having 1 to 18 carbon atoms.

[0022] q is an integer from 0 to 18.

[0023] For some optional structures of the compound of formula I, R2 is or

[0024] Wherein, p is an integer of 1, t is an integer of 1 (R3 replaces one hydrogen atom on the benzene ring, and R4 replaces one hydrogen atom on the five-membered heterocyclic ring), R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C10 alkoxy, C1-C10 alkyl, C1-C10 carboxylalkyl, C1-C10 aminoalkyl, C1-C10 hydroxyalkyl, or

[0025] R5 is a carboxyl-substituted alkoxy group having 1-10 carbon atoms, a carboxyl-substituted alkylamino group having 1-10 carbon atoms, or an amino-substituted alkylamino group having 1-10 carbon atoms; R6 is hydrogen, carboxyl, or a C1-C10 alkyl group. Z is independently O, S, or NR8.

[0026] R7 and R8 are each independently hydrogen, an alkyl group having 1 to 10 carbon atoms, a carboxyalkyl group having 1 to 10 carbon atoms, an aminoalkyl group having 1 to 10 carbon atoms, or a hydroxyalkyl group having 1 to 10 carbon atoms.

[0027] q is an integer from 0 to 10.

[0028] For some optional structures of the compound of formula I, R2 is or

[0029] Wherein, p is an integer of 2, t is an integer of 2 (R3 replaces 2 hydrogen atoms on the benzene ring, and R4 replaces 2 hydrogen atoms on the five-membered heterocyclic ring), R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C10 alkoxy, C1-C10 alkyl, C1-C10 carboxylalkyl, C1-C10 aminoalkyl, C1-C10 hydroxyalkyl, or

[0030] R5 is an alkoxy group substituted with a carboxyl group having 1 to 10 carbon atoms, an alkylamino group substituted with a carboxyl group having 1 to 10 carbon atoms, or an alkylamino group substituted with an amino group having 1 to 10 carbon atoms. R6 is hydrogen or a carboxyl group.

[0031] Z is independently O, S or NR8.

[0032] R7 and R8 are each independently hydrogen, an alkyl group having 1 to 10 carbon atoms, a carboxyalkyl group having 1 to 10 carbon atoms, an aminoalkyl group having 1 to 10 carbon atoms, or a hydroxyalkyl group having 1 to 10 carbon atoms.

[0033] q is an integer from 0 to 10.

[0034] For some optional structures of the compound of formula I, R3 is

[0035] or

[0036] The compound of formula I has the following resonance structures. The following resonance structures are four resonance structures of the same compound:

[0037]

[0038] Among them, R1, R2, n, X - The definition of is the same as that in formula I.

[0039] For the compound of formula I, when n is 1, 2, or 3, the compound may have the following structure:

[0040]

[0041] in,

[0042] R1, R2, n, X - The definition of is the same as that in formula I.

[0043] Another aspect of the present invention is to provide a method for preparing the compound of formula I. The specific preparation process is as follows:

[0044] First, a keto intermediate W is synthesized, and then W is condensed with the corresponding aldehyde or other condensing agent S to obtain an enone structure compound T. Finally, W and T are catalyzed by boron trifluoride ether to generate the corresponding near-infrared second-region fluorescent dye;

[0045] The synthetic route of the dye is as follows:

[0046]

[0047] Where: R1, R2, n, X - The definition of is the same as that in formula I.

[0048] Taking n=3 as an example, the preparation method of the compound of formula I is specifically described, which specifically comprises the following steps:

[0049]

[0050] (1) Synthesis of W: Dissolve the corresponding salicylaldehyde in benzene, add equal amounts of glacial acetic acid and piperidine, and react at 90°C for 1.5 h. Then add equal amounts of cyclohexenone, raise the temperature to 140°C, and react for 72 h. The product is obtained by silica gel column chromatography.

[0051] (2) Synthesis of T: W was dissolved in methanol, and a 5-fold aqueous solution of NaOH was added under ice-cooling, and the mixture was stirred at room temperature overnight. The product was separated and purified by silica gel column chromatography.

[0052] (3) Synthesis of dye: W and T were placed in a reaction flask under argon protection. Boron trifluoride etherate was added as solvent and the reaction was refluxed overnight. After the reaction was completed, the mixture was cooled to room temperature, poured into a large amount of water, extracted, dried, and separated by column chromatography.

[0053] Where: R1, R2, X - The definition of is the same as that in formula I.

[0054] In the present invention, the carboxyl-substituted alkoxy group refers to an alkyl group in which a carboxyl group is substituted for an alkoxy group, the carboxyl-substituted alkylamino group refers to an alkyl group in which a carboxyl group is substituted for an alkylamino group, and the amino-substituted alkylamino group refers to an alkyl group in which an amino group is substituted for an alkylamino group.

[0055] The carboxyalkyl group refers to an alkyl group substituted by a carboxyl group, the aminoalkyl group refers to an alkyl group substituted by an amino group, and the hydroxyalkyl group refers to an alkyl group substituted by a hydroxy group.

[0056] The beneficial effects of the present invention are as follows: 1) The compounds provided by the present invention have absorption wavelengths greater than 800 nanometers and emission wavelengths exceeding 1000 nanometers, exhibiting very large Stokes chemical shifts. These compounds can be used for near-infrared optical imaging in the second region, avoiding drawbacks such as dye self-absorption, biological autofluorescence, and light scattering. 2) These compounds can absorb near-infrared light and generate heat through excited-state relaxation. For example, in Example 9, the photothermal conversion efficiency can reach 60%, showing potential applications in photothermal therapy. 3) After absorbing near-infrared light and reaching the excited state of the dye, these compounds reach the triplet state through intersystem crossing, sensitizing oxygen. For example, in Example 9, the singlet oxygen quantum yield can reach 20%, showing potential applications in photodynamic therapy. 4) Through their large conjugated planes and inherent positive charge, these compounds form an amphiphilic structure that can target cardiolipin, a specific phospholipid in the mitochondrial inner membrane, and affect the function of cardiolipin-binding protein, thereby triggering the apoptotic pathway and demonstrating strong anti-cancer activity, showing significant application prospects in chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 is the absorption spectrum of the compound in Example 9.

[0058] Figure 2 is the absorption spectrum of the compound in Example 11.

[0059] Figure 3 is the luminescence spectrum of the compound in Example 9.

[0060] Figure 4 is the luminescence spectrum of the compound in Example 11.

[0061] Figure 5 The compound of Example 9 was subjected to light irradiation and temperature increase.

[0062] Figure 6 The killing effect of the compound in Example 9 on different cells. DETAILED DESCRIPTION

[0063] The specific embodiments of the present invention are described in detail below in conjunction with the technical solutions and drawings.

[0064] The present invention is illustrated but not limited by the following examples in which all parts and percentages are by weight unless otherwise indicated.

[0065] Example 1

[0066]

[0067] 3.0 g of 4-dimethyl salicylaldehyde was dissolved in 100 mL of benzene, and 3 mL of glacial acetic acid and 3 mL of piperidine were added. The mixture was stirred at 90 ° C for 1.5 h. 1.5 mL of cyclohexenone was added, and the temperature was raised to 140 ° C and stirred for 72 h. The product was separated and purified by silica gel column chromatography. HRMS (M+H + ):244.1332.

[0068]

[0069] Dissolve 3.0 g of 4-diethylsalicylaldehyde in 100 mL of benzene, add 3 mL of glacial acetic acid and 3 mL of piperidine, and stir at 90°C for 1.5 h. Add 1.4 mL of cyclopentenone, raise the temperature to 140°C, and stir for 72 h. The product is purified by silica gel column chromatography; HRMS (M+H+): 258.3560.

[0070]

[0071] Dissolve 3.0 g of 4-diethylsalicylaldehyde in 100 mL of benzene, add 3 mL of glacial acetic acid and 3 mL of piperidine, and stir at 90°C for 1.5 h. Add 1.5 mL of cycloheptenone, raise the temperature to 140°C, and stir for 72 h. The product is purified by silica gel column chromatography; HRMS (M+H+): 286.1922.

[0072]

[0073] The synthesis method of W4-W11 refers to the synthesis of W1, and the product structure is identified by mass spectrometry.

[0074] Example 2

[0075]

[0076] S1 was synthesized with reference to the following literature: Nat Methods. 12, 2015, 244–250, Bioorg. Med. Chem. 2004, 12, 4749–4759, Angew. Chem. Int. Ed. 2017, 56, 15545–15549.

[0077] The specific operations are as follows:

[0078] Under an ice bath, add 3 mL of POCl₃ to 5 mL of DMF and stir at room temperature for 30 minutes. Dissolve 2 g of 8-hydroxyjulolidine in 3 mL of DMF and add dropwise to the solution. Heat to 60°C and react for 6 hours. Pour the reaction solution into ice water and adjust the pH to neutral with sodium bicarbonate. A large amount of solid will precipitate. S1 is further isolated and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1).

[0079]

[0080] The synthesis methods of S2-S7 were based on the synthesis of S1, and the product structures were identified by mass spectrometry.

[0081] Example 3

[0082]

[0083] Dissolve W1 (0.02 mmol), benzaldehyde (0.02 mmol), and sodium hydroxide (0.2 mmol) in 100 mL of methanol and stir at room temperature overnight. The product was purified by column chromatography, HRMS (M+H+): 332.1645.

[0084] Example 4

[0085]

[0086] Dissolve W1 (0.02 mmol), thiophene carboxaldehyde (0.02 mmol), and sodium hydroxide (0.2 mmol) in 100 mL of methanol and stir at room temperature overnight. The product was purified by column separation, HRMS (M+H+): 338.1209.

[0087] Example 5

[0088]

[0089] Dissolve W1 (0.02 mmol), 1-naphthaldehyde (0.02 mmol), and sodium hydroxide (0.2 mmol) in 100 mL of methanol and stir at room temperature overnight. The product was purified by column chromatography, HRMS (M+H+): 332.1645.

[0090] Example 6

[0091]

[0092] Dissolve W2 (0.02 mmol), 1-naphthaldehyde (0.02 mmol), and sodium hydroxide (0.2 mmol) in 100 mL of methanol and stir at room temperature overnight. The product was purified by column chromatography, HRMS (M+H+): 409.2042.

[0093] T5-T12 were synthesized by referring to the above method.

[0094] Example 7

[0095]

[0096] Dissolve W4 (0.02 mmol), benzaldehyde (0.02 mmol), and sodium hydroxide (0.2 mmol) in 100 mL of methanol and stir at room temperature overnight. The product was purified by column chromatography, HRMS (M+H+): 346.1802.

[0097] Example 8

[0098] The synthesis methods of other intermediates T14-T34 are as follows: Example 7. The raw materials used are intermediate W and aldehydes containing different substituents R2. All raw materials are commercially available or synthesized according to existing technologies.

[0099] Example 9

[0100]

[0101] W1 (0.02 mmol), T1 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 555.2642.

[0102] Example 10

[0103]

[0104] W1 (0.02 mmol), T5 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 583.2955.

[0105] Example 11

[0106]

[0107] W2 (0.02 mmol), T5 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 611.3268.

[0108] Example 12

[0109]

[0110] W2 (0.02 mmol), T4 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 661.3425.

[0111] Example 13

[0112]

[0113] W2 (0.02 mmol), T6 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 711.3581.

[0114] Example 14

[0115]

[0116] W3 (0.02 mmol), T7 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 659.3268.

[0117] Example 15

[0118]

[0119] W2 (0.02 mmol), T8 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 656.3119.

[0120] Example 16

[0121]

[0122] W2 (0.02 mmol), T8 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 654.3690.

[0123] Example 17

[0124]

[0125] W2 (0.02 mmol), T10 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 689.2373.

[0126] Example 18

[0127]

[0128] W2 (0.02 mmol), T11 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 639.3581.

[0129] Example 19

[0130]

[0131] W2 (0.02 mmol), T12 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 627.3217.

[0132] Example 20

[0133]

[0134] W2 (0.02 mmol), T13 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 597.3112.

[0135] Example 21

[0136]

[0137] W4 (0.02 mmol), T13 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 583.2955.

[0138] Example 22

[0139]

[0140] W5 (0.02 mmol), T5 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 625, 3425.

[0141] Example 23

[0142]

[0143] W5 (0.02 mmol), T14 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 639.3581.

[0144] Example 24

[0145]

[0146] W6 (0.02 mmol), T15 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 551.2329.

[0147] Example 25

[0148]

[0149] W7 (0.02 mmol), T16 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 579.2642.

[0150] Example 26

[0151]

[0152] W8 (0.02 mmol), T17 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 607.2955.

[0153] Example 27

[0154]

[0155] W9 (0.02 mmol), T18 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation, HRMS (M+): 635.3268.

[0156] Example 28

[0157]

[0158] W10 (0.02 mmol), T19 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 639.2853.

[0159] Example 29

[0160]

[0161] W11 (0.02 mmol), T20 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 665.3486.

[0162] Example 30

[0163]

[0164] W2 (0.02 mmol), T21 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed evenly and refluxed overnight. The product was purified by column separation. HRMS (M + ):700.3534.

[0165] Example 31

[0166]

[0167] W2 (0.02 mmol), T22 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 727.3894.

[0168] Example 32

[0169]

[0170] W2 (0.02 mmol), T23 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 655.3166.

[0171] Example 33

[0172]

[0173] W2 (0.02 mmol), T24 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 699.3065.

[0174] Example 34

[0175]

[0176] W2 (0.02 mmol), T25 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 655.3166.

[0177] Example 35

[0178]

[0179] W2 (0.02 mmol), T26 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 647.3080.

[0180] Example 36

[0181]

[0182] W2 (0.02 mmol), T27 (0.02 mmol), and boron trifluoride ether solution (10 mL) were mixed and refluxed overnight. The product was purified by column separation, HRMS (M+): 701.2797.

[0183] Example 37

[0184]

[0185] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0186] Example 38

[0187]

[0188] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0189] Example 39

[0190]

[0191] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0192] Example 40

[0193]

[0194] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0195] Example 41

[0196]

[0197] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0198] Example 42

[0199]

[0200] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0201] Example 43

[0202]

[0203] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0204] Example 44

[0205]

[0206] The synthesis method was similar to that of Example 33, and the product structure was identified by mass spectrometry.

[0207] Example 45

[0208]

[0209] Dissolve 100 mg of Dye 3 in 50 mL of ethanol. Dissolve 20 times the sodium chloride in an appropriate amount of water and add to the dye. Stir overnight at 25°C. Filter to remove insoluble salts, spin dry the solvent, and separate by column chromatography. HRMS (M+): 611.3268.

[0210] Example 46

[0211]

[0212] Dissolve 100 mg of Dye 3 in 50 mL of dichloromethane. Add 20 times the volume of sodium hexafluorophosphate to the dye and stir overnight at 25°C. Remove insoluble salts by filtration, spin dry the solvent, and filter the mixture. HRMS (M+): 611.3268.

[0213] Example 47

[0214] Dye absorption spectrum test method: Taking the compounds in Example 9 and Example 11 as an example, the dye was prepared into a 10mM mother solution with DMSO, and its absorption spectrum in different solvents was tested respectively, and the test concentration was 3μM. Figure 1 It can be seen that the maximum absorption wavelength of the compound in Example 9 is 850nm (in dichloromethane). Figure 2 It can be seen that the maximum absorption wavelength of the compound in Example 11 is 870 nm (in dichloromethane), which has reached the near-infrared region.

[0215] Example 48

[0216] Dye fluorescence spectrum test method: Take the compounds dye1 and dye3 in Example 9 and Example 11 as examples, use DMSO to prepare a 10mM stock solution of the dye, use an 850nm light source to excite, and collect the fluorescence spectra in different solvents respectively. The test concentration is 10μM. Figure 3 It can be seen that the maximum emission wavelength of compound dye 1 in Example 9 is 960 nm (in dichloromethane). Figure 4 It can be seen that the maximum emission wavelength of compound dye3 in Example 11 is 961 nm (in dichloromethane), and covers the second near-infrared region.

[0217] Example 49

[0218] Dye light temperature increase experiment: Taking compound dye1 in Example 9 as an example, the dye was prepared into a 10mM stock solution with DMSO. The dye stock solution was dispersed in deionized water to prepare different concentrations (0, 5, 10, 20, 50μM). 1mL of this aqueous solution was placed in a cuvette. The cuvette was irradiated with an 808nm laser at a power of 1W, and the temperature change was recorded using a near-infrared thermal imager. Figure 5 It can be seen that the dye has a high light-to-heat conversion ability, and 50 μM can increase the water temperature by 40 °C.

[0219] Example 50

[0220] Dye cell killing effect: Using compound dye 1 from Example 9 as an example, the dye was prepared into a 10 mM stock solution using DMSO. The dye was then diluted to different concentrations in culture medium and used to culture different cells. After 24 hours, cell viability was measured using the MTT assay, and the half-inhibitory concentration (IC) was calculated for different cell types. 50 .from Figure 6It can be seen that dye1 shows high killing ability against all cancer cells. For example, the half-inhibitory concentration for HeLa cells is 1μM. At the same time, it can be seen that the dye has weak killing ability against normal cells, which can be used to distinguish normal cells from cancer cells.

[0221] Example 51

[0222] The spectral properties of compounds dye4, dye6, dye11, dye13, dye15, dye19, dye20, dye23, dye25, dye27, dye29, dye31, dye33, and dye35 in dichloromethane are given in the following table. The test conditions are the same as those in Example 47 and Example 48.

[0223]

[0224]

[0225] The above specific examples illustrate the embodiments of the present invention. Those skilled in the art will appreciate the other advantages and benefits of the present invention from the disclosure herein. The present invention may also be implemented or applied through various other specific embodiments, and the details in this specification may be modified or altered based on different viewpoints and applications without departing from the spirit of the present invention.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof: in, n is independently 1, 2 or 3; When n is 1, 2, or 3, the structural fragment They are R1 is independent of each other or N, when R1 is N, it connects to the carbon atoms on the left and right adjacent positions of the carbon atom on the substituted benzene ring to form a six-membered ring structure; R2 is or wherein p is an integer of 1-5, t is an integer of 1-3, and R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C20 alkoxy, C1-C20 alkyl, C1-C20 carboxylalkyl, C1-C20 aminoalkyl, C1-C20 hydroxyalkyl, or R5 is a C1-C10 carboxyl-substituted alkoxy group, a C1-C10 carboxyl-substituted alkylamino group, or a C1-C10 amino-substituted alkylamino group; R6 is hydrogen, carboxyl, or a C1-C20 alkyl group; Z is independently O, S or NR8; R7 and R8 are each independently hydrogen, C1-C18 alkyl, C1-C18 carboxyalkyl, C1-C18 aminoalkyl or C1-C18 hydroxyalkyl; m and q are each independently an integer from 0 to 18; X - and Y - is an anion selected from PF 6- 、BF4 - 、Cl - Br - , I - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - CF3SO3 - ; The total positive charge is equal to X - The total amount of negative charge; The total positive charge is equal to Y - The total amount of negative charge.

2. The compound according to claim 1, characterized in that p is an integer of 1-2, t is an integer of 1-2, R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C20 alkoxy, C1-C20 alkyl, C1-C20 carboxylalkyl, C1-C20 aminoalkyl, C1-C20 hydroxyalkyl, or R5 is a C1-C10 carboxyl-substituted alkoxy group, a C1-C10 carboxyl-substituted alkylamino group, or a C1-C10 amino-substituted alkylamino group; R6 is hydrogen, carboxyl, or a C1-C20 alkyl group; Z is independently O, S or NR8; R7 and R8 are each independently hydrogen, C1-C18 alkyl, C1-C18 carboxyalkyl, C1-C18 aminoalkyl or C1-C18 hydroxyalkyl; q is an integer from 0 to 18.

3. The compound according to claim 1, characterized in that p is an integer of 1, t is an integer of 1, R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C10 alkoxy, C1-C10 alkyl, C1-C10 carboxylalkyl, C1-C10 aminoalkyl, C1-C10 hydroxyalkyl, or R5 is a C1-C10 carboxyl-substituted alkoxy group, a C1-C10 carboxyl-substituted alkylamino group, or a C1-C10 amino-substituted alkylamino group; R6 is hydrogen, carboxyl, or a C1-C20 alkyl group; Z is independently O, S or NR8; R7 and R8 are each independently hydrogen, C1-C18 alkyl, C1-C18 carboxyalkyl, C1-C18 aminoalkyl or C1-C18 hydroxyalkyl; q is an integer from 0 to 10.

4. The compound according to claim 1, characterized in that p is an integer of 2, t is an integer of 2, R3 and R4 are each independently hydrogen, hydroxyl, carboxyl, amino, azido, nitro, halogen, phosphate, C1-C10 alkoxy, C1-C10 alkyl, C1-C10 carboxylalkyl, C1-C10 aminoalkyl, C1-C10 hydroxyalkyl, or R5 is a C1-C10 carboxyl-substituted alkoxy group, a C1-C10 carboxyl-substituted alkylamino group, or a C1-C10 amino-substituted alkylamino group; R6 is hydrogen, carboxyl, or a C1-C20 alkyl group; Z is independently O, S or NR8; R7 and R8 are each independently hydrogen, C1-C18 alkyl, C1-C18 carboxyalkyl, C1-C18 aminoalkyl or C1-C18 hydroxyalkyl; q is an integer from 0 to 10.

5. A compound of formula I or a pharmaceutically acceptable salt thereof: in, n is independently 1, 2 or 3; When n is 1, 2, or 3, the structural fragment They are R1 is independent of each other or N, when R1 is N, it connects to the carbon atoms on the left and right adjacent positions of the carbon atom on the substituted benzene ring to form a six-membered ring structure; m is an integer from 0 to 18; X - and Y - is an anion selected from PF 6- 、BF4 - 、Cl - Br - , I - 、NO3 - 、SO4 2- 、ClO4 - 、CH3COO - 、CH3SO3 - CF3SO3 - ; described The total positive charge is equal to X - The total amount of negative charge; The total positive charge is equal to Y - The total amount of negative charge; R2 is or 6. The method for preparing the compound according to claim 1, wherein The following steps are involved: First, a ketone intermediate W is synthesized, and then W is subjected to a condensation reaction with a corresponding aldehyde or condensing agent to obtain a ketene structure compound T. Finally, W and T are reacted under the catalysis of boron trifluoride ether to produce a compound of formula I. The reaction formula is as follows: Where: R1, R2, n, X - The definition is the same as that in formula I of claim 1; When n is 1, 2, or 3, the structural fragment They are structure They are 7. The use of the compound according to claim 1, characterized in that: The compound is used as a near-infrared fluorescent dye for preparing optical imaging drugs in the near-infrared first region and / or infrared second region.

8. The use of the compound according to claim 1, characterized in that: The compound is used for preparing medicine for inducing cancer cell apoptosis.

9. The use of the compound according to claim 1, characterized in that: The compound is used for preparing chemotherapy drugs.

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