Novel near-infrared two-region thioxanthene dye, J-aggregate of novel near-infrared two-region thioxanthene dye, and preparation method and application of J-aggregate

Abstract

The invention relates to the technical field of near-infrared second-region organic dyes, in particular to a novel near-infrared second-region thioxanthene dye, a J-aggregate thereof, a preparation method of the J-aggregate and application of the J-aggregate. The preparation method of the dye comprises the following steps: (1) adding inorganic alkali or organic alkali into a two-neck bottle under the protection of nitrogen; the preparation method comprises the following steps: adding m-hydroxythiophenol or m-aminothiophenol dissolved in an organic solvent into a two-neck flask, stirring at room temperature for 0-2 hours, adding a parent CL-1, and reacting at 25-100 DEG C for 0.5-12 hours; and (2) cooling to room temperature, pouring the mixture into water, extracting with dichloromethane or ethyl acetate, washing with a saturated saline solution, drying with anhydrous sodium sulfate, concentrating under vacuum, and purifying with silica gel column chromatography. The novel near-infrared two-region thioxanthene dye synthesized by the invention is good in photophysical property and singlet oxygen generation performance, has a good photo-thermal heating effect, and has the potential of tumor diagnosis and treatment as an excellent photo-thermal reagent.

Description

Technical Field

[0001] The present invention relates to the technical field of near-infrared zone II organic dyes, in particular to a novel near-infrared zone II thioxanthene dye, its J-aggregate, and a preparation method and application thereof. Background Art

[0002] The development of fluorophores and photothermal agents in the second near-infrared window (NIR-II, 1000-1700 nm) is an emerging field in the field of bioimaging and tumor diagnosis. Compared with traditional bioimaging within 1000 nm, near-infrared second-region bioimaging has many advantages, such as high spatiotemporal resolution, deep tissue penetration, low tissue light absorption / scattering and autofluorescence. Currently, most near-infrared second-region organic dyes mainly include polymethylene, benzobithiadiazole (BBTD) and xanthene-based derivatives. Although BBTD derivatives have made encouraging progress in bioimaging and tumor diagnosis / therapy, they are still limited by complex chemical synthesis, absorption wavelength (λ abs <900nm) and emission wavelength (λ em <1100nm). Indocyanine green (ICG), as a typical polymethine dye, has been approved by the US FDA for clinical bioimaging, but its absorption / emission peaks are located at 780 / 822nm, and the emission tail region can only reach 1000nm. Therefore, ICG can only be excited by 808nm laser, which seriously limits its application in high-resolution bioimaging of deep tissues. We know that the penetration depth and resolution of bioimaging depend not only on the emission wavelength, but also on the excitation wavelength. Compared with excitation wavelengths less than 1000nm, bioimaging with an excitation wavelength of 1064nm has higher imaging contrast in deep tissues. Therefore, there is an urgent need to develop new organic fluorophores that have both absorption and fluorescence in the near-infrared second window.

[0003] Peripheral hyperthermia (PTT) is a non-invasive treatment modality that induces apoptosis and necrosis of tumor cells through local hyperthermia with precise spatiotemporal control. Although PTT is highly effective in killing tumors, it also has some inherent drawbacks, such as the physical limitation of optical penetration into tissues and the maximum permissible exposure (MPE) when the excitation wavelength is less than 1000 nm. For example, when biological tissue is irradiated with 808 nm laser, the maximum permissible exposure is 0.33 W / cm 2 This severely limits the therapeutic efficacy of PTT for deep-seated tumors. Furthermore, with the exception of a few polymers, the excitation wavelength of most examples is below 1000 nm due to inherent defects. Therefore, the simultaneous implementation of organic dye-based near-infrared second-region fluorescence bioimaging and PTT at an excitation wavelength of 1064 nm is of great significance.

[0004] Generally, dyes with large conjugated structures exhibit significant red-shifted absorption / emission wavelengths due to strong intramolecular charge transfer (ICT) effects. However, dyes with large planar π-conjugated structures are limited by aggregation-induced quenching in aqueous solutions, which hinders bright near-infrared (NIR) II bioimaging. Supramolecular self-assemblies composed of ordered molecular building blocks hold great promise for constructing high-performance functional materials. These self-assemblies, formed by intermolecular interactions such as dipole-dipole, π-π interactions, and hydrogen bonds, often exhibit properties distinct from their individual units, exceeding simply the sum of their components. J-aggregates, characterized by a head-to-tail sliding molecular arrangement, represent a unique class of ordered supramolecular assemblies. J-type exciton coupling within J-aggregates leads to significantly red-shifted absorption, significantly enhancing the penetration depth of light in living systems and facilitating near-infrared optical imaging and phototherapy. Furthermore, compared to monomers or amorphous aggregates, J-aggregates exhibit higher molar absorption coefficients (ε), enhanced emission brightness, and improved photothermal effects, offering further advantages for phototherapy. Therefore, constructing J-aggregates based on near-infrared region II organic dyes has high potential for clinical application. Summary of the Invention

[0005] The present invention provides a novel near-infrared second-zone thioxanthene dye, its J-aggregate, and its preparation method and application. The novel dye has good photothermal warming effect and has the potential to be used as an excellent photothermal agent for tumor diagnosis and treatment, thus solving the problems existing in the prior art.

[0006] One of the technical solutions adopted by the present invention is:

[0007] A new type of near-infrared second-zone thioxanthene dye is provided, and its chemical structure is as follows:

[0008] Designated as SXH, where X=O or NH.

[0009] Furthermore, the near-infrared second region thioxanthene dye is SOH or SNH2, and the chemical structural formulas are:

[0010]

[0011] The second technical solution adopted by the present invention is:

[0012] A method for preparing the aforementioned near-infrared second-region thioxanthene dye is provided, comprising the following steps:

[0013] (1) Under nitrogen protection, a certain amount of inorganic base or organic base is added to a two-necked flask; m-hydroxythiophenol or m-aminothiophenol dissolved in 50 mL of organic solvent is added to the two-necked flask, stirred at room temperature for 0 to 2 hours, and then a certain amount of parent CL-1 is added, and the reaction is carried out at 25 to 100°C for 0.5 to 12 hours;

[0014] The structural formula of the parent CL-1 is as follows:

[0015] The chemical name is:

[0016] (E)-4-(((E)-2-chloro-3-((6-(diethylamino)-2,3-dihydro-1H-xanthen-4-yl)methylene)cyclohex-1-en-1-yl)methylene)-6-(diethylamino)-1,2,3,4-tetrahydroxanthylium perchlorate;

[0017] (2) After the reaction is completed, cool to room temperature, pour the mixture into water, extract with dichloromethane or ethyl acetate (100 mL × 3), wash with saturated brine (100 mL × 3), dry over anhydrous sodium sulfate, and concentrate under vacuum, then purify by silica gel column chromatography to obtain;

[0018] Wherein, the molar amount of the inorganic base or organic base, and the molar amount of m-hydroxythiophenol or m-aminothiophenol in step (1) are 1 to 5 times the molar amount of the parent CL-1.

[0019] Furthermore, the molar amount of the parent CL-1 is 1.34 mmol, and the molar amount of the inorganic base or organic base, and the molar amount of m-hydroxythiophenol or m-aminothiophenol are all 1.34 to 6.7 mmol, respectively.

[0020] Furthermore, in step (1), the inorganic base is sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, cesium carbonate or ammonia; the organic base is diethylamine, triethylamine, pyridine, imidazole, piperidine, sodium methoxide or sodium ethoxide; and the organic solvent in step (1) is 50 mL of dichloromethane, chloroform, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, methanol, ethanol or tetrahydrofuran.

[0021] Furthermore, the chromatographic conditions of the silica gel column chromatography in step (2) are: eluent: CH2Cl2 / CH3OH, 100 / 1 to 10 / 1 for elution.

[0022] Furthermore, the target product SOH obtained by the above preparation steps had a yield of 88% and an HRMS-ESI detection value of 482.21634. The target product SNH2 obtained by the above preparation steps had a yield of 85% and an HRMS-ESI detection value of 481.23081.

[0023] Furthermore, the reaction formula for synthesizing SOH and SNH2 in the above preparation steps is as follows:

[0024]

[0025] The third technical solution adopted by the present invention is:

[0026] The aforementioned near-infrared second region thioxanthene dyed J-aggregate is provided, which is prepared by the following preparation steps:

[0027] S1 completely dissolved the near-infrared second-region thioxanthene dye and Pluronic@F127 in tetrahydrofuran;

[0028] S2: Slowly add the dye solution from step S1 to the beaker containing PBS while stirring. After the addition is complete, continue stirring at room temperature for a certain period of time.

[0029] S3: Place the beaker in an ultrasonicator, sonicate at 180W, remove excess tetrahydrofuran, and then filter the solution through a 0.22 μm polyvinylidene fluoride (PVDF) filter using a syringe to obtain.

[0030] Furthermore, the amount of Pluronic@F127 used in step S1 is 1 to 100 times the amount of the near-infrared second-region thioxanthene dye used, the volume amount of tetrahydrofuran used is 1 to 10 times the amount of the dye used, and the volume amount of PBS used in step S2 is 1 to 50 times the amount of the dye used.

[0031] Furthermore, in step S1, the amount of the near-infrared second region thioxanthene dye used is 2 mg;

[0032] The amount of Pluronic@F127 used is 2-200 mg, and the amount of tetrahydrofuran used is 2-20 mL. In step S2, the amount of PBS used is 10-100 mL, and the stirring time at room temperature is half an hour. In step S3, the ultrasonic treatment is performed for 2 hours.

[0033] Furthermore, Pluronic F127 in S1 is a thermosensitive hydrogel, which is a non-ionic triblock copolymer composed of polyethylene glycol (PEG) and polypropylene glycol (PPO), with a specific structure of PEG-PPO-PEG, and is an existing commercial product.

[0034] The fourth technical solution adopted by the present invention is:

[0035] Provided is the use of the J-aggregate of the aforementioned near-infrared second-region thioxanthene dye as an angiographic contrast agent.

[0036] Furthermore, the angiographic contrast agent is a near-infrared second-zone fluorescent angiographic contrast agent or a near-infrared second-zone photoacoustic imaging angiographic contrast agent.

[0037] Furthermore, the J aggregate prepared as above was used for angiography, and the resolution of blood vessels observed in near-infrared zone II fluorescence angiography was as high as 0.19 mm, and the resolution of blood vessels observed in near-infrared zone II photoacoustic angiography was as high as 0.06 mm.

[0038] The fifth technical solution adopted by the present invention is:

[0039] Provided is the use of the J-aggregate of the aforementioned near-infrared second-region thioxanthene dye as a photo-diagnostic agent.

[0040] Furthermore, the optical diagnostic and therapeutic agent is a photodynamic therapy agent or a photothermal therapy agent.

[0041] Furthermore, the J aggregates prepared above are used for optical diagnosis and treatment of tumors, with a tumor cure rate of up to 100%.

[0042] Beneficial effects of the present invention:

[0043] 1. The near-infrared second-zone thioxanthene dye synthesized by the present invention has excellent photophysical properties and singlet oxygen generation performance, and has good photothermal warming effect, and has the potential to be used as an excellent photothermal agent for tumor diagnosis and treatment.

[0044] 2. The J-aggregate prepared by using the near-infrared second zone thioxanthene dye of the present invention has good biocompatibility and can be detected by 1064 nm laser (1 W / cm 2 , 10min), it effectively inhibited cell proliferation; in vivo experiments showed that it can perform fluorescence angiography and photoacoustic angiography of blood vessels with high resolution, and has a good anti-tumor effect when performing PTT after being irradiated in tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 To test the singlet oxygen generation ability of the two near-infrared second-region thioxanthene dyes synthesized by the present invention;

[0046] Figure 2 The light-induced temperature rise curves of the two near-infrared second-region thioxanthene dyes synthesized by the present invention;

[0047] Figure 3 Absorption spectrum (left) and fluorescence spectrum (right) of the J-aggregate J-SOH NPs of the present invention;

[0048] Figure 4The absorption spectrum (left) and fluorescence spectrum (right) of the J-aggregate J-SNH2NPs of the present invention;

[0049] Figure 5 To investigate the dark toxicity of U87-MG cells incubated with different concentrations of J-SOH NPs and J-SNH2NPs;

[0050] Figure 6 To investigate the phototoxicity of U87-MG cells incubated with different concentrations of J-SOH NPs and J-SNH2NPs, a 1064 nm laser (1 W / cm 2 , 10min);

[0051] Figure 7 After the animals were injected with J-SOH NPs, fluorescence images of blood vessels near the tumor were obtained under 980 nm excitation;

[0052] Figure 8 Photoacoustic images of whole-body angiography in mice 30 minutes after tail vein injection of J-SNH2NPs under 1064 nm excitation;

[0053] Figure 9 The changes in tumor volume of mice within 14 days after different treatments. DETAILED DESCRIPTION

[0054] In order to clearly illustrate the technical features of this solution, the present invention is described in detail below through specific implementation methods in conjunction with the accompanying drawings.

[0055] In the present invention, the equipment and raw materials used can be purchased from the market or are commonly used in the art. The methods in the following examples are all conventional methods in the art unless otherwise specified.

[0056] Example 1 Synthesis of target product SOH

[0057] The novel near-infrared second-region thioxanthene dye SOH was synthesized as follows:

[0058]

[0059] Under nitrogen, 0.162 g (1.60 mmol) of the organic base triethylamine was added to a two-necked flask. 0.202 g (1.60 mmol) of m-hydroxythiophenol dissolved in 50 mL of dichloromethane was also added to the flask. After stirring at room temperature for 2 hours, 1 g (1.34 mmol) of the parent compound CL-1 was added, and the mixture was allowed to react at 30°C for 6 hours. After completion of the reaction, the mixture was cooled to room temperature and poured into water. The mixture was extracted with dichloromethane (100 mL x 3), washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by silica gel column chromatography (eluent: CH2Cl2 / CH3OH, 100 / 1 to 10 / 1) to obtain 0.69 g of the desired product, SOH, in an 88% yield. The HRMS-ESI value was 482.21634.

[0060] Example 2

[0061] The method for synthesizing the novel near-infrared second-region thioxanthene dye SOH followed the same preparation steps as in Example 1, except that the precursor CL-1 was added and the reaction was carried out at 50°C for 4 hours. The target product SOH was obtained with an 85% yield and an HRMS-ESI value of 482.21634.

[0062] Example 3

[0063] The method for synthesizing the novel near-infrared second-region thioxanthene dye SOH followed the same preparation steps as in Example 1, except that the precursor CL-1 was added and the reaction was carried out at 90°C for 1 hour. The target product SOH was obtained with an 82% yield and an HRMS-ESI value of 482.21634.

[0064] Example 4 Synthesis of target product SNH2

[0065] The novel near-infrared second-region thioxanthene dye SNH2 was synthesized as follows:

[0066]

[0067] Under nitrogen, 0.162 g (1.60 mmol) of triethylamine was added to a two-necked flask. 0.199 g (1.60 mmol) of m-aminothiophenol dissolved in 50 mL of dichloromethane was also added to the flask. After stirring at room temperature for 2 hours, 1 g (1.34 mmol) of the parent compound CL-1 was added, and the mixture was allowed to react at 30°C for 6 hours. After completion of the reaction, the mixture was cooled to room temperature and poured into water. The mixture was extracted with dichloromethane (100 mL x 3), washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, and concentrated under vacuum. The mixture was then purified by silica gel column chromatography (eluent: CH2Cl2 / CH3OH, 100 / 1 to 10 / 1) to obtain 0.66 g of the desired product, SNH2, in an 85% yield. The HRMS-ESI value was 481.23081.

[0068] Example 5J Aggregate Preparation

[0069] 2 mg of SOH prepared in Example 1 and 2 mg of Pluronic@F127 were completely dissolved in 2 mL of tetrahydrofuran. A beaker containing 10 mL of PBS was prepared and the dye solution was slowly added dropwise to the beaker while stirring. After the addition was complete, stirring was continued at room temperature for half an hour. The beaker was then placed in an ultrasonicator and sonicated at 180 W for two hours. Excess tetrahydrofuran was removed using a rotary evaporator, and the solution was then filtered through a 0.22 μm polyvinylidene fluoride (PVDF) filter using a syringe to obtain J-aggregate J-SOH NPs.

[0070] The same method was used to replace SOH with SNH2 prepared in Example 4 to obtain J aggregates J-SNH2NPs.

[0071] 1. Photophysical properties of SOH and SNH2

[0072] The following tests were performed on the SOH and SNH2 prepared in Example 1 and Example 4.

[0073] (1) Absorption and emission spectra

[0074] SOH or SNH2 was dissolved in dimethyl sulfoxide, methanol, acetonitrile, N,N-dimethylformamide, tetrahydrofuran, dichloromethane, and chloroform, respectively, to prepare a 5 mM stock solution. The concentration was determined using a UV-visible-near-infrared spectrophotometer and a near-infrared steady-state transient fluorescence phosphorescence spectrometer. The test concentration was 10 μM, with fluorescence excitation wavelengths of 808 nm and 980 nm.

[0075] (2) Determination of relative fluorescence quantum yield

[0076] Dissolve SOH or SNH2 in anhydrous dimethyl sulfoxide, methanol, anhydrous acetonitrile, N,N-dimethylformamide, anhydrous tetrahydrofuran, anhydrous dichloromethane, and chloroform, respectively. Adjust the maximum absorbance of the dye at 808nm to around 0.1OD, and set 5 concentration points in sequence. Use origin integration to obtain the fluorescence peak area of ​​each concentration under 808nm excitation, then use the area as the ordinate and the corresponding absorbance value as the abscissa to obtain the slope of the linear fit. Using IR-26 (Φ = 0.05% in 1,2-dichloroethane) as the standard, the fluorescence quantum yield Φ of SOH in different solvents is calculated by the following formula F .

[0077]

[0078] where the subscripts S and X denote the standard sample and the test sample, respectively. R is the slope of the integrated emission intensity versus absorbance. η is the refractive index of the solvent.

[0079] The above results are shown in Tables 1 and 2.

[0080] Table 1 Photophysical properties of SOH

[0081]

[0082] a Abbreviation: λ abs : Absorption maximum, λ em : maximum emission value, Φ F : fluorescence quantum yield, ε: molar extinction coefficient

[0083] Table 2 Photophysical properties of SNH2

[0084]

[0085]

[0086] a Abbreviation: λ abs : Absorption maximum, λ em : emission maximum value, Φ F : fluorescence quantum yield, ε: molar extinction coefficient

[0087] (3) Singlet oxygen generation performance measurement

[0088] The singlet oxygen generation test of SOH or SNH2 uses N'N-dimethylformamide as solvent and DPBF as singlet oxygen capture reagent for measurement. The detection mechanism is that the singlet oxygen generated by the dye quickly oxidizes DPBF, causing its maximum absorption peak (415nm) to attenuate. In order to eliminate the inner filter effect, the absorbance of SOH or SNH2 at 808nm is adjusted to 0.20OD, and then 30μL of DPBF solution (1mg / mL, DMF) is added to the above 2mL solution, and irradiated with 808nm laser at 1.0W / cm 2 The absorption spectrum data of DPBF were collected by monitoring for 300 s at 30 s intervals under the power of . Figure 1 .

[0089] (4) Photothermal performance measurement

[0090] The SOH or SNH2 stock solution was diluted to 10 μM with phosphate buffer solution. The thermocouple was used to measure the thermal conductivity of the SOH or SNH2 stock solution under 808 nm laser irradiation (1 W / cm 2 , 10min) heating curve.

[0091] To evaluate the photothermal performance of SOH and SNH2, the photothermal 2 , 10min) under the stimulation of different concentrations of SOH and SNH2. Figure 2 As shown, SOH at a concentration of 10 μM heated to over 48°C under illumination, while SNH2 heated to 49°C after 10 minutes of illumination at a concentration of 10 μM, demonstrating that both SOH and SNH2 exhibited excellent photothermal heating effects. The photothermal conversion efficiency for SOH was 48%, while that for SNH2 was 50%, indicating that both SOH and SNH2 can serve as excellent photothermal agents for tumor diagnosis and treatment.

[0092] 2. Absorption and fluorescence spectra of J-SOH NPs and J-SNH2NPs

[0093] See also Figure 3-4 The J-SOH NPs and J-SNH2NPs prepared above showed a sharp absorption peak near 1100nm and displayed an ultra-strong fluorescence signal at 1150nm.

[0094] 3. Cytotoxicity Experiment

[0095] U87-MG cells were seeded at an appropriate density into 96-well plates. After 24 hours of proliferation, different concentrations of J-SOH NPs or J-SNH2 NPs were added to MEM. After incubation at 37°C for 24 hours, the old medium was removed and replaced with fresh medium containing CCK-8 solution (CCK-8 solution / culture medium = 1:10, 100 μL per well). The cells were incubated at 37°C for another 1 hour. The absorbance at 450 nm was then measured on a microplate reader, and cell viability was calculated using the following formula.

[0096]

[0097] OD (drug added): absorbance values ​​for wells containing cells, culture medium, CCK-8 solution, and drug solution; OD (blank): absorbance values ​​for wells containing culture medium and CCK-8 solution but no cells; OD (0 drug added): absorbance values ​​for wells containing cells, culture medium, CCK-8 solution but no drug solution. Each experiment was performed in parallel six times.

[0098] The phototoxicity and dark toxicity of J-SOH NPs and J-SNH2NPs were quantitatively analyzed using U87-MG cancer cells by CCK-8 assay. Figure 5 As shown in Figure 2, after incubation with different concentrations of J-SOH NPs and J-SNH2NPs for 24 h, the cell viability exceeded 90%, indicating that both J-SOH NPs and J-SNH2NPs have good biocompatibility. 2 When cells were irradiated for 10 min, both J-SOH NPs and J-SNH2NPs effectively inhibited cell proliferation, and the survival rate of U87-MG was less than 10% at a concentration of 10 μg / mL ( Figure 6 ).

[0099] IV. In vivo experiments

[0100] (1) Construction of tumor mouse model

[0101] All mice were housed under SPF-grade conditions at a temperature of 26°C, a humidity of 50%, and adequate water and food. U87-MG cancer cells (1×10 5 ) was injected subcutaneously into the right abdomen of each mouse to establish a U87-MG tumor xenograft mouse model. 3 The mice were randomly divided into six groups: PBS, PBS+L, J-SOH NPs, J-SOH NPs+L, J-SNH2NPs, and J-SNH2NPs+L. The mice in the laser irradiation group received 1064 nm laser (1 W / cm 2) were irradiated for 10 min and the experiment was performed. Tumor volume was measured every two days using a caliper and the calculation formula was as follows:

[0102]

[0103] V represents the tumor volume of the mouse, a represents the longest diameter of the tumor area, and b represents the width of the longest diameter area of ​​the tumor.

[0104] (2) Near-infrared second-region in vivo fluorescence imaging

[0105] Prepare tumor volume at 100mm 3 J-SOH NPs were injected into the tail veins of approximately 100,000 tumor-bearing mice. The injected tumor-bearing mice were then imaged using a near-infrared (NIR) II in vivo fluorescence imager with an excitation wavelength of 980 nm and filters of 1100LP, 1150LP, and 1200LP, respectively.

[0106] After administration of J-SOH NPs, the fluorescence imaging was performed at the same position using 980 nm laser excitation. Figure 7 As shown, the fluorescence signal at 30 min is much stronger than that at 0 min. The FWHM of the blood vessel at cross-section line "1" is 0.26 mm, and the FWHM of the blood vessel at cross-section line "2" is 0.22 and 0.19 mm, respectively. The FWHM of the blood vessel at cross-section line "3" is 0.19 mm, demonstrating that J-SOH NPs are capable of high-resolution fluorescence imaging of blood vessels.

[0107] Similarly, the administration of J-SNH2NPs for the above imaging can also perform fluorescence imaging of blood vessels with high resolution.

[0108] (3) In vivo photoacoustic imaging in the near-infrared region II

[0109] Several tumor-bearing mice with appropriate tumor volumes were prepared, and J-SOH NPs or J-SNH2NPs were injected into the tail vein. The injected tumor-bearing mice were imaged using a near-infrared second-zone in vivo photoacoustic imager under 1064 nm laser excitation.

[0110] After administration of J-SNH2NPs, the photoacoustic signal intensity of whole-body angiography of mice was detected. Figure 8, the photoacoustic signal at 30 min is much stronger than that at 0 min. In order to analyze the photoacoustic imaging ability of J-SNH2NPs for angiography, four areas were selected in the vascular photoacoustic imaging diagram, cross-sectional lines 1, 2, and 3 in the left tumor vascular imaging and cross-sectional line 4 in the right body vascular imaging. The full width at half maximum (FWHM) of the blood vessels at cross-sectional line 1 are 0.32 and 0.19 mm, respectively. The FWHM of the blood vessels at cross-sectional line 2 are 0.28 and 0.31 mm, respectively; the FWHM of the blood vessels at cross-sectional line 3 are 0.28 and 0.26 mm, respectively; the FWHM of the blood vessels at cross-sectional line 4 are 0.27, 0.28, 0.06, and 0.20 mm, respectively. Therefore, J-SNH2NPs can perform photoacoustic angiography of blood vessels with high resolution.

[0111] Similarly, after administering J-SOH NPs, the photoacoustic signal intensity of whole-body angiography in mice was detected, and the photoacoustic signal at 30 minutes was much stronger than that at 0 minutes; and J-SOH NPs can also perform photoacoustic angiography of blood vessels with high resolution.

[0112] (4) In vivo phototherapy

[0113] Tumor-bearing mice were randomly divided into six groups (PBS, PBS+L, J-SOH NPs, J-SOH NPs+L, J-SNH2NPs, J-SNH2NPs+L). Mice in the laser irradiation group received 1064 nm laser (1 W / cm 2 The tumor volume of the mice was measured every two days, and the mice were euthanized after 14 days of treatment.

[0114] To evaluate the antitumor effect of J-SOH NPs in vivo, BALB / c mice bearing subcutaneous U87-MG xenografts were divided into six groups: (1) PBS group, mice were intratumorally injected with PBS; (2) J-SOH NPs group, mice were only given J-SOH NPs; (3) J-SNH2NPs group, mice were only given J-SNH2NPs; (4) PBS+L group, mice were intratumorally injected with PBS for 0.5 h and then irradiated (1064 nm, 1 W / cm 2 , 10 min); (5) J-SOH NPs+L group, mice were administered with J-SOH NPs and then irradiated with laser (1064 nm, 1 W / cm 2 , 10 min); (6) J-SNH2NPs+L group, mice were administered with J-SNH2NPs and then irradiated with laser (1064 nm, 1 W / cm 2 , 10min).

[0115] During the treatment period, the tumor volume was measured every 2 days to evaluate the photothermal therapy effect of each group. Figure 9As shown in the figure, the tumor volumes of the PBS group, PBS+L group, J-SOH NPs group, and J-SNH2NPs group gradually increased over time. After 14 days, the tumor volumes of the four groups increased by 8-10 times, indicating that light, J-SOH NPs, and J-SNH2NPs could not inhibit tumor growth. However, the tumor volumes of the J-SOH NPs or J-SNH2NPs light-exposed groups were significantly inhibited and cured, with the tumor volume increasing from 100 mm 3 Reduced to 0mm left and right 3 The results showed that J-SOH NPs and J-SNH2NPs had good anti-tumor effects after laser irradiation and PTT in tumor cells.

[0116] The above describes in detail the novel near-infrared second-region thioxanthene dyes, their J-aggregates, and their preparation methods and applications provided by the present invention. The aforementioned specific embodiments are not intended to limit the scope of the present invention. Those skilled in the art will appreciate that any modifications or variations to the embodiments of the present invention fall within the scope of the present invention.

[0117] Any matters not described in detail in the present invention are well-known technologies to those skilled in the art.

Claims

1. A novel near-infrared second-region thioxanthene dye, characterized in that: Its chemical structure is as follows: wherein X=O or NH.

2. A method for preparing the near-infrared second region thioxanthene dye according to claim 1, characterized in that: The steps are as follows: (1) Under nitrogen protection, add an inorganic base or an organic base into a two-necked flask; add m-hydroxythiophenol or m-aminothiophenol dissolved in an organic solvent into the two-necked flask, stir at room temperature for 0 to 2 hours, then add the parent CL-1 and react at 25 to 100°C for 0.5 to 12 hours; (2) After the reaction is completed, the mixture is cooled to room temperature, and then poured into water, extracted with dichloromethane or ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under vacuum, and then purified by silica gel column chromatography to obtain; Wherein, the molar amount of the inorganic base or organic base, and the molar amount of m-hydroxythiophenol or m-aminothiophenol in step (1) are 1 to 5 times the molar amount of the parent CL-1.

3. The method for preparing near-infrared second region thioxanthene dye according to claim 2, characterized in that: In step (1), the inorganic base is sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, cesium carbonate or ammonia water; the organic base is diethylamine, triethylamine, pyridine, imidazole, piperidine, sodium methoxide or sodium ethoxide; and the organic solvent in step (1) is 50 mL of dichloromethane, chloroform, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, methanol, ethanol or tetrahydrofuran.

4. The method for preparing near-infrared second region thioxanthene dye according to claim 2, wherein The chromatographic conditions of the silica gel column chromatography in step (2) are: eluent: CH2Cl2 / CH3OH, 100 / 1 to 10 / 1 for elution.

5. A J-aggregate dyed with near-infrared second region thioxanthene as claimed in claim 1, characterized in that: The following preparation steps are adopted: S1 completely dissolved the near-infrared second-region thioxanthene dye and Pluronic@F127 in tetrahydrofuran; S2: Slowly add the dye solution from step S1 to the beaker containing PBS while stirring. After the addition is complete, continue stirring at room temperature for a certain period of time. S3 was placed in an ultrasonicator and ultrasonicated at 180W to remove excess tetrahydrofuran, and then the solution was filtered through a 0.22 μm polyvinylidene fluoride filter to obtain the product.

6. The J-aggregate according to claim 5, characterized in that In step S1, the amount of Pluronic@F127 used is 1 to 100 times the amount of the near-infrared second region thioxanthene dye used, the volume of tetrahydrofuran used is 1 to 10 times the amount of the near-infrared second region thioxanthene dye used, the volume of PBS used in step S2 is 1 to 50 times the amount of the near-infrared second region thioxanthene dye used, and the stirring time at room temperature is half an hour; in step S3, the ultrasonic treatment is performed for 2 hours.

7. Use of the J-aggregate according to claim 5 or 6 as an angiographic contrast agent.

8. The use according to claim 7, characterized in that The angiographic contrast agent is a near-infrared second-zone fluorescent angiographic contrast agent or a near-infrared second-zone photoacoustic imaging angiographic contrast agent.

9. Use of the J-aggregate according to claim 5 or 6 as a phototherapeutic agent.

10. The use according to claim 9, characterized in that The optical diagnostic and therapeutic agent is a photodynamic therapy agent or a photothermal therapy agent.

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