Raman image probe molecule with photothermal conversion capability and preparation and preparation method thereof

Abstract

The invention relates to a Raman image probe molecule with photothermal conversion capability and a preparation and a preparation method thereof. The Raman image probe takes thiophene coupled benzo-bis-thiadiazole as a parent nucleus, and has specific Raman shift and good photothermal conversion efficiency. In a gathering state, after being irradiated by near-infrared light (700-900 nm), the material can generate a characteristic Raman scattering signal, can convert light energy into heat energy, and has a good photothermal conversion property. The nano preparation is prepared from distearoyl phosphatidyl ethanolamine-polyethylene glycol and a derivative thereof or albumin or a polyethylene glycol block copolymer and the Raman image probe, and has Raman scattering signal and photothermal conversion performance. The prepared nano preparation can generate strong characteristic Raman scattering signals under the irradiation of near-infrared light and can convert light energy into heat energy. The nano preparation can be applied to tumor living body Raman spectrum imaging and photothermal therapy, and realizes tumor diagnosis and treatment integrated precise therapy.

Description

Technical Field

[0001] This invention belongs to the fields of molecular imaging and pharmaceutical formulation technology, and relates to a class of Raman imaging probe molecules with photothermal conversion capabilities, their formulations, and preparation methods. In particular, it relates to small-molecule Raman imaging probes for in vivo Raman spectroscopy imaging and photothermal therapy, their formulations, and preparation methods. Background Technology

[0002] Photothermal therapy for tumors utilizes localized high temperatures to directly kill tumor cells. It boasts advantages such as high specificity and minimal side effects, showing great promise in tumor treatment. However, during photothermal therapy, it is difficult to accurately locate primary and metastatic tumors using only visual inspection. Therefore, developing therapeutic molecules and their formulations that combine imaging diagnostics and photothermal therapy functions to improve the precision of photothermal therapy and reduce damage to surrounding normal tissues is crucial for enhancing the safety and efficacy of photothermal therapy.

[0003] Raman spectroscopy imaging, with its fingerprint-like characteristics and high resolution, holds great promise for applications in tumor imaging. Raman spectroscopy imaging technology is based on the molecular Raman scattering effect. In the interaction between an electromagnetic field (light) and molecules, when the energy of the scattered photon is equal to that of the incident photon, the process is called Rayleigh scattering, a typical elastic scattering process. When the energy of the scattered photon is lower than that of the incident photon, some energy is transferred to the molecular vibrational energy level, resulting in a redshift; this process is called Stokes Raman scattering. Conversely, when the molecule transfers vibrational energy to the photon, causing the energy of the scattered photon to be higher than that of the incident photon, resulting in a blueshift, this is called anti-Stokes Raman scattering. Both of these types of scattering belong to inelastic scattering. Generally, the Stokes Raman scattering signal of a molecule is stronger than the anti-Stokes signal. Therefore, the Stokes portion of the spectrum is mainly used for Raman scattering spectroscopy.

[0004] But every 10 7 Only one photon out of every 100 photons will undergo Raman scattering. The extremely weak spontaneous Raman scattering signal is difficult to detect, necessitating the use of surface-enhanced Raman scattering (SERS) to amplify the Raman scattering signal. SERS uses metallic materials (gold, silver, copper, etc.) as a substrate and enhances the Raman scattering signal of reporter molecules coated on the substrate surface through physical or chemical enhancement mechanisms. While SERS-based probes have enabled intraoperative Raman spectroscopy-guided photothermal therapy for tumors, the poor biocompatibility of inorganic substrates used in SERS hinders the clinical application of these probes.

[0005] Because tumor photothermal therapy requires precise imaging to locate lesions, and existing SERS in vivo imaging agents need to be prepared with metal as the base material, there are issues with biosafety and difficulty in in vivo metabolism. Furthermore, existing technologies cannot obtain Raman imaging probes that simultaneously have Raman scattering signal emission without substrate enhancement and good photothermal conversion performance. Summary of the Invention

[0006] Based on the shortcomings of existing technologies and related research foundations, the purpose of this invention is to provide a Raman imaging probe molecule with good photothermal conversion capabilities.

[0007] The present invention also aims to provide a method for synthesizing Raman imaging probe molecules with good photothermal conversion capabilities.

[0008] Another object of the present invention is to provide the Raman imaging probe formulation having good photothermal conversion capability.

[0009] Another object of the present invention is to provide a method for preparing the preparation.

[0010] The objective of this invention is achieved through the following scheme: This invention designs a class of Raman imaging probe molecules with photothermal conversion properties. These are small molecule organic compounds capable of generating Raman scattering signals and possessing photothermal conversion properties, exhibiting an 894 cm⁻¹ diameter. -1 and 1264 cm -1 The characteristic peaks of the Raman shift, in their aggregated state, do not require surface enhancement with a metal substrate and generate strong Raman scattering signals upon near-infrared light irradiation. The molecular structure of the Raman imaging probe is based on thiophene-coupled benzobisthiadiazole as the parent core, and its general structural formula I is as follows: General Formula I In Formula I, R1, R2, R3, and R4 are -H, -F, -Cl, -Br, -I, -OH, -CHO, -NH2, -COOH, and -C, respectively. n H 2n+1 , -OC n H 2n+1 , -(0CH2) n CH3, substituted or unsubstituted Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution , substituted or non-substituted , substituted or non-substituted

[0011] The Raman image probe with the light-heat conversion capability has characteristic peaks of 894 cm -1 and 1264 cm -1 Raman shift, and can be used for in-vivo Raman spectrum imaging and Raman image probe with light-heat conversion capability, which realizes in-vivo Raman spectrum imaging, overcomes the problems that the existing SERS in-vivo imaging agent needs to be prepared by taking metal as a base material, and has biological safety and difficulty in in-vivo metabolism; meanwhile, the Raman image probe and the preparation thereof also have high light-heat conversion capability, and can be used for in-vivo photothermal therapy.

[0012] The application provides a preparation method of the above-mentioned Raman image probe with light-heat conversion capability, and the preparation method comprises the following steps: Synthesis route 1: 4,7-dibromobenzodithiazole and α or β substituted tributyl (thiophene-2-yl) stannane are used as raw materials, a Stille reaction is performed, and the target product is obtained, and the reaction formula of the synthesis route 1 is as follows: (1) wherein R1, R2, R3 and R4 are -H, -F, -Cl, -Br, -I, -OH, -CHO, -NH2, -COOH, -C n H 2n+1 , , -OC n H 2n+1 , , -(OCH2) n CH3, substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , and the like. In the above reaction formula (1), the reagent a is tetrakis triphenylphosphine palladium, 1,4-dioxane, and the reaction temperature is 105 DEG C.

[0013] Alternatively, synthesis route 2: the β-substituted 4,7-dithiophenyl benzodithiazole obtained by the synthesis route 1 is used as raw material, bromination and Suzuki reaction are performed, and the target product is obtained, and the reaction formula of the synthesis route 2 is as follows: (2) wherein R1, R2, R3, R4 are -H, -F, -Cl, -Br, -I, -OH, -CHO, -NH2, -COOH, -C n H 2n+1 , , -OC n H 2n+1 , , -(OCH2) n CH3, substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , substituted or non-substituted , and the like; In the above reaction formula (2), The reagent a used is N-bromosuccinimide (NBS), chloroform, N,N-dimethylformamide (DMF), and the reaction condition is ice bath; The reagent b is 2 M K2CO3, tetraphenylphosphine palladium, aryl, heteroaryl boronic acid or aryl, heteroaryl pinacol borate, 1,4-dioxane, and the reaction temperature is 105 DEG C.

[0014] The application also provides the use of the above-mentioned Raman image probe with light-heat conversion capability for the preparation of in-vivo Raman spectrum image and photothermal therapy.

[0015] The Raman image probe provided by the application is a small-molecule Raman image probe, which, in an aggregated state, does not need metal substrate enhancement, generates strong Raman scattering signals under 700-900 nm near-infrared light irradiation, can be used for in-vivo Raman spectrum image, and can convert light energy into heat energy, and can be used as a preparation of in-vivo photothermal therapy.

[0016] The small-molecule Raman image probe and the preparation thereof have high efficient light-heat conversion capability, can realize the diagnosis and treatment integration of in-vivo tumor Raman spectrum image guided photothermal therapy, and have good clinical application prospect.

[0017] In the present application, the Raman imaging probe is characterized in that the thienyl-coupled benzodithiazole is used as a mother nucleus, has an aggregation-induced Raman scattering enhancement property, and can generate a strong Raman scattering signal for in vivo Raman spectral imaging under near-infrared light irradiation without a metal substrate surface enhancement in an aggregated state. At the same time, the Raman imaging probe and its preparation also have high efficient photothermal conversion capability and can be used for in vivo photothermal therapy.

[0018] The present application provides a preparation of the Raman imaging probe with the above-mentioned photothermal conversion capability, comprising: (I) a micellar preparation of the Raman imaging probe according to any one of the above, wherein the preparation contains the Raman imaging probe and distearoyl phosphatidyl ethanolamine-polyethylene glycol and its derivatives.

[0019] The mass ratio of the Raman imaging probe to distearoyl phosphatidyl ethanolamine-polyethylene glycol and its derivatives is 1:0.2-1:50; preferably, the mass ratio is 1:0.2-1:10.

[0020] The distearoyl phosphatidyl ethanolamine-polyethylene glycol and its derivatives are one or a mixture of several of distearoyl phosphatidyl ethanolamine-polyethylene glycol, distearoyl phosphatidyl acetic amide-polyethylene glycol-amino, distearoyl phosphatidyl acetic amide-polyethylene glycol-maleimide, and distearoyl phosphatidyl acetic amide-polyethylene glycol-thiol. The molecular weight of the polyethylene glycol is between 500-5000. Preferably, the distearoyl phosphatidyl ethanolamine-polyethylene glycol and its derivatives are distearoyl phosphatidyl acetic amide-polyethylene glycol 2000.

[0021] The preparation method of the micellar preparation containing the Raman imaging probe comprises the following steps: (1) dissolving distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000 and the Raman imaging probe in chloroform, dichloromethane, tetrahydrofuran or a mixed solvent thereof; (2) adding the mixed solution in (1) dropwise into water and diluting ten times; (3) ultrasonically treating the mixed solution in (2) for a certain time; (4) stirring the mixed solution in (3) in a fume hood for a certain time; (5) filtering the mixed solution in (4) through a 0.22 µm filter membrane to prepare a micelle.

[0022] Preferably, the solvent in step (1) is chloroform.

[0023] Preferably, the ultrasonic treatment time in step (3) is 2-10 min.

[0024] Preferably, the stirring time in step (4) is 8-12 h.

[0025] (ii) The albumin nano-preparation containing any one of the above Raman image probes, wherein the preparation contains the Raman image probe and albumin, wherein, The mass ratio of the Raman image probe to albumin is 1:0.2-1:50; preferably, the mass ratio is 1:0.2-1:10. The albumin is one or a mixture of several of human serum albumin, recombinant human serum albumin and bovine serum albumin.

[0026] The preparation method of the albumin nano-preparation containing the Raman image probe, comprising the following steps: (1) Dissolving albumin in water; (2) Dissolving the Raman image probe in methanol, ethanol, propylene glycol, chloroform, dichloromethane, tetrahydrofuran or a mixture of the above solvents; (3) Adding the organic solution of step (2) to the aqueous solution of step (1) and mixing uniformly; (4) Stirring the mixture obtained in step (3) at room temperature for 1-8 h.

[0027] Preferably, the concentration of the albumin aqueous solution in step (1) is 0.1%-5% (w / v), preferably 0.5%-2% (w / v).

[0028] Preferably, the ratio of the organic solvent in step (2) to water in step (1) is 1:40-1:60.

[0029] Preferably, the stirring time in step (4) is 2-6 h.

[0030] (iii) The nano-particle preparation containing any one of the above Raman image probes, wherein the preparation contains the Raman image probe and a polyethylene glycol-based block copolymer, wherein, The mass ratio of the Raman image probe to the polyethylene glycol-based block copolymer is 1:0.2-1:50; preferably, the mass ratio is 1:0.2-1:10. The polyethylene glycol block copolymer is one or a mixture of several of polyethylene glycol-poly(lactic-co-glycolic acid) block copolymer, polyethylene glycol-poly(caprolactone) block copolymer, polyethylene glycol-poly(lactic acid) block copolymer, and polyethylene glycol-poly(carbonate) block copolymer. The molecular weight of the polyethylene glycol block copolymer ranges from 2000 to 60000, and the molecular weight of PEG ranges from 1000 to 5000. Preferably, the polyethylene glycol block copolymer is polyethylene glycol-poly(lactic-co-glycolic acid) block copolymer (PEG-PLGA), the molecular weight of PLGA is 50000, LA:GA = 50:50, and the molecular weight of PEG is 2000.

[0031] The preparation method of the PEG-PLGA nano-preparation containing the Raman imaging probe comprises the following steps: (1) dissolving the Raman imaging probe and PEG-PLGA in a certain proportion in methanol, ethanol, propylene glycol, chloroform, dichloromethane, tetrahydrofuran or a mixture thereof; (2) under the conditions of ultrasonic and ice water bath, mixing the organic solution of step (1) in pure water; (3) stirring the mixture obtained in step (2) at room temperature for 1-8 h to evaporate the organic solvent; (4) freeze-drying the nanoparticle solution obtained in step (3) to obtain the desired nanoparticles.

[0032] Preferably, the mass ratio of the Raman imaging probe to PEG-PLGA in step (1) is 1:0.2-1:10.

[0033] Preferably, the ratio of the organic solvent to pure water in step (2) is 1:10-1:20.

[0034] Preferably, the stirring time in step (3) is 2-6 h.

[0035] The above-mentioned any one kind of nano-preparation loaded with Raman imaging probe is irradiated by near-infrared light, and the in-vivo Raman spectrum image is obtained without the need of metal substrate surface enhancement; preferably, the wavelength of the excitation light source is 785 nm and 830 nm laser.

[0036] The above-mentioned any one kind of nano-preparation loaded with Raman imaging probe is irradiated by near-infrared light, and the light energy can be converted into heat energy, and the photo-thermal conversion efficiency is good; preferably, the wavelength of the excitation light source is 808 nm laser.

[0037] The prepared nano-preparation of the application has improved water solubility compared with free Raman image probe, and meets the requirements of drug dosage for Raman spectrum image and photothermal therapy. The mouse experiment results show that the nano-preparation of the Raman probe can be accumulated in the tumor site after intravenous injection. Under the irradiation of 785 nm or 830 nm laser, the in-situ tumor focus and metastatic focus can be clearly imaged during the operation. The experimental results of the tumor photothermal therapy guided by the in-situ Raman spectrum image show that the photothermal conversion effect of the nano-preparation in the body under the irradiation of 808 nm laser can effectively kill tumor cells.

[0038] The application meets the needs of tumor visualization and photothermal therapy by designing and preparing a compound molecule into a diagnosis and treatment integrated nano-preparation, which has Raman spectrum image and photothermal conversion performance. BRIEF DESCRIPTION OF DRAWINGS

[0039] Figure 1 : is the chemical structural formula of compound 3 in (a) of Example 1; (b) is the Raman scattering spectrum of compound 3 in 95% water: 5% THF mixed solution under the excitation of 785 nm excitation light, the concentration is 10 μM; (c) is the temperature change curve of compound 3 in 95% water: 5% THF mixed solution under the excitation of 808 nm excitation light, the concentration is 50 μM, and the laser power is 1 W / cm 2 ; Figure 2 : is the chemical structural formula of compound 5 in (a) of Example 2; (b) is the Raman scattering spectrum of compound 5 in 95% water: 5% THF mixed solution under the excitation of 785 nm excitation light, the concentration is 10 μM; (c) is the temperature change curve of compound 5 in 95% water: 5% THF mixed solution under the excitation of 808 nm excitation light, the concentration is 50 μM, and the laser power is 1 W / cm 2 ; Figure 3 : is the chemical structural formula of compound 7 in (a) of Example 3; (b) is the Raman scattering spectrum of compound 7 in 95% water: 5% THF mixed solution under the excitation of 785 nm excitation light, the concentration is 10 μM; (c) is the temperature change curve of compound 7 in 95% water: 5% THF mixed solution under the excitation of 808 nm excitation light, the concentration is 50 μM, and the laser power is 1 W / cm 2 ; Figure 4: (a) Chemical structure of compound 9 in Example 4; (b) Raman scattering spectrum of compound 9 in 95% water: 5% THF mixed solution under excitation of 785 nm excitation light, concentration of 10 μΜ; (c) Temperature change curve of compound 9 in 95% water: 5% THF mixed solution under excitation of 808 nm excitation light, concentration of 50 μΜ, laser power of 1 W / cm 2 ; Figure 5 : (a) Chemical structure of compound 11 in Example 5; (b) Raman scattering spectrum of compound 11 in 95% water: 5% THF mixed solution under excitation of 785 nm excitation light, concentration of 10 μΜ; (c) Temperature change curve of compound 11 in 95% water: 5% THF mixed solution under excitation of 808 nm excitation light, concentration of 50 μΜ, laser power of 1 W / cm 2 ; Figure 6 : (a) Particle size distribution of BBTPPRO micelles prepared from compound 11 in Example 6; (b) Raman scattering spectrum of BBTPPRO micelles under excitation of 785 nm excitation light, concentration of 10 μΜ; (c) Temperature change curve of BBTPPRO micelles under excitation of 808 nm excitation light, concentration of 50 μΜ, laser power of 1 W / cm 2 ; Figure 7 : (a) Visible light photos and corresponding Raman spectrum image pictures of orthotopic tumor and metastatic tumor of CT26-Luc orthotopic tumor model mice after tail vein administration of BBTPPRO micelles (7.5 mg / kg) for 24 h, position 1 is normal tissue of intestinal tract, position 2 is orthotopic tumor, position 3 is metastatic tumor; (b) Raman scattering signals of different positions of intestinal tract in Figure a; (c) Hematoxylin-eosin staining pictures of orthotopic tumor and metastatic tumor and corresponding Raman spectrum image pictures; (d) Temperature change curve of tumor under 808 nm (1 W / cm 2 ) irradiation after tail vein administration of BBTPPRO micelles (7.5 mg / kg) for 24 h of CT26-Luc orthotopic tumor model mice; (e) Infrared thermal imaging pictures of mouse tumor under excitation of 808 nm laser for different time; Figure 8: Survival curves for tumor-bearing mice in Example 6. Mice bearing the CT26-Luc orthotopic tumor model underwent different treatments; the control group received no treatment, the nanoparticle administration group received BBTPPROmicelle (7.5 mg / kg) via tail vein only, and the laser irradiation group had tumors identified by visual inspection and were irradiated with an 808 nm laser for 8 minutes at a power of 1 W / cm². 2 Twenty-four hours after mice in the nanoparticle + laser group were administered BBTPPRO micelle (7.5 mg / kg) via tail vein, photothermal therapy was performed on the tumor site using optical fiber under Raman spectroscopy imaging guidance. The in situ tumor was irradiated with an 808 nm laser for 5 minutes, and the metastatic tumor was irradiated with an 808 nm laser for 8 minutes, with a power of 1 W / cm². 2 In the nanoparticle + laser group, no tumor recurrence was observed in mice within 70 days, while the other three groups of mice were not cured and all died within 35 days (n = 5); p <0.01, compared with the nanoparticle + laser group; Figure 9 (a) Particle size distribution of albumin nanoparticles (BBTPRO NPs) prepared from compound 9 in Example 7; (b) Raman scattering spectrum of albumin nanoparticles BBTPRO NPs under 785 nm excitation at a concentration of 10 μM; (c) Temperature change curve of albumin nanoparticles BBTPRO NPs under 808 nm excitation at a concentration of 50 μM and a laser power of 1 W / cm². 2 ; Figure 10 (a) Particle size distribution of nanoparticles (BBTPPRO NPs) prepared from compound 11 in Example 8; (b) Raman scattering spectrum of nanoparticles (BBTPPRO NPs) under 785 nm excitation at a concentration of 10 μM; (c) Temperature change curve of nanoparticles (BBTPPRO NPs) under 808 nm excitation at a concentration of 50 μM and a laser power of 1 W / cm². 2 . Detailed Implementation

[0040] The present invention will be further described below with reference to embodiments, but is not limited thereto.

[0041] Example 1: Synthetic reaction formula for Raman imaging probe 4,7-bis(4-(2-propyl)thiophen-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 3): The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesis was carried out in the following steps: (1) 4,7-Dibromo-benzo[l,2-c:4,5-c']bis[l,2,5]thiadiazole (compound 1, 0.6 g, 1.70 mmol) and tetrakis(triphenylphosphine)palladium (196 mg, 0.16 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (25 mL) and tributyl((2-propyl)thiophen-2-yl)stannane (compound 2, 2.36 g, 5.68 mmol) was added under nitrogen. The flask was flushed with nitrogen (x 3); (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation and the residue was purified by column chromatography using dichloromethane / petroleum ether (1:1, v / v) as eluent to give the target product compound 3 (376 mg, yield 50%) as a blue solid. 1 H NMR (600MHz, CDCl3) δ 8.84 (s, 2H), 7.32 (d, J = 12.0 Hz, 2H), 2.76 (t, J = 7.6 Hz,4H), 1.80 (dq, J = 14.8, 7.4Hz, 4H), 1.05 (t, J = 7.3 Hz, 6H). 13 C NMR (151MHz, CDCl3) δ 151.05, 144.09, 137.43, 134.12, 126.49,113.72, 32.81, 23.93,14.13. MALDI-TOF MS Calcd for: C 20 H 17 N4S4 - ([M-H] - ): 442.0409. Found: 442.1567. Compound 3 ( Figure 1 a) Characteristic peaks of Raman shift of 894 cm -1 and 1264 cm -1 were detected upon excitation at 785 nm. Figure 1 b) Upon excitation at 808 nm with a power density of 1 W / cm 2 , the temperature of a 95% water:5% THF mixture solution was observed to increase by 9.3 °C within 10 min ( Figure 1 c).

[0042] The obtained compound 3 has both Raman spectral imaging and photothermal conversion performance.

[0043] Example 2: The reaction formula for synthesizing the Raman imaging probe 4,7-bis(4-(2- ethylhexyl)thiophene-2-yl)benzo[l,2-c:4,5-c']bis[l,2,5]thiadiazole (compound 5, BBT) is as follows: Reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; Synthesis was carried out as follows: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and tributyl(4-(2-ethylhexyl)thiophene-2-yl)stannane (compound 4, 1.38 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x3); (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, and the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain blue solid product BBT (0.27 g, yield 56%). 1 H NMR (400 MHz, CDCl3)δ 8.67 (s, 2H), 7.28 (s, 2H), 2.66 (d, J = 6.8 Hz, 4H), 1.84 – 1.71(m, 2H),1.45 – 1.31 (m, 16H), 1.05 – 0.87 (m, 12H). 13C NMR (151 MHz, CDCl3) δ150.55, 142.29, 136.50, 133.95, 126.50, 113.00, 39.79, 33.97, 31.90, 28.30,25.07, 22.51, 13.60, 10.30. MALDI-TOF MS Calcd for: C 30 H 37 N4S4 - ([M-H] - ):582.1924. Found: 582.2315.

[0044] BBT( Figure 2a) Upon excitation at 785 nm, the characteristic peaks of Raman shift of 894 cm -1 and 1264 cm -1 can be detected. Figure 2 b) Upon excitation at 808 nm with a power density of 1 W / cm 2 , it can be seen that the temperature of the compound 5 in a 95% water: 5% THF mixed solution increased by 11.6 °C within 10 min ( Figure 2 c).

[0045] The obtained compound 5 has both Raman spectral imaging and photothermal conversion performance.

[0046] Example 3: Synthesis reaction formula of Raman imaging probe 4,7-bis((4-(2-ethylhexyl)-5-(4-phenyl)thiophene-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 7): The reaction reagents and conditions of step a are: NBS, chloroform, DMF, ice bath; was synthesized by the following steps: (1) NBS (0.14 g, 0.78 mmol) was added dropwise to a chloroform / DMF (10 mL, 1:1, v / v) mixed solution of BBT (0.22 g, 0.38 mmol) at room temperature; (2) The mixture was stirred in an ice bath under nitrogen protection for 10 min, then quenched with ice water, and the aqueous phase was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate after washing with water and saturated brine; (3) After removing dichloromethane under reduced pressure, the residue was purified by silica gel chromatography eluted with ethyl acetate / petroleum ether (1:120, v / v) to obtain the desired product compound 6 (106 mg, yield 38%) as a blue solid. 1 H NMR (400MHz, CDCl3) δ 8.57 (s, 2H), 2.63 (d, J = 6.8 Hz, 4H), 1.84 – 1.70 (m, 2H),1.46 – 1.33 (m, 16H), 0.98-0.92 (m, 12H). 13C NMR (151 MHz, CDCl3) δ 149.89,141.62, 136.38, 133.35, 117.31, 111.74, 39.39, 33.21, 31.91, 28.18,25.16,22.54, 13.60, 10.32. MALDI-TOF MS Calcd for: C 30 H 37 Br2N4S4 + ([M+H] + ): 742.0269.Found: 742.0602.

[0047] Step b reaction reagent and condition: phenylboronic acid, tetrakis(triphenylphosphine)palladium, 2M K2CO3, 1,4-dioxane, 105 °C; Synthesized by the following steps: (1) Compound 6 (112 mg, 0.152 mmol), phenylboronic acid (48 mg, 0.38 mmol) and tetrakis(triphenylphosphine)palladium (16 mg, 0.015 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (10 mL) was added by syringe under nitrogen and the resulting solution was degassed by vacuum. 2M K2CO3solution (0.24 mL) was added under nitrogen; (3) The mixture was further degassed by vacuum and then stirred at 105 °C for 4 h; (4) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane. The organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate; (5) The product was purified by silica gel chromatography with ethyl acetate / petroleum ether (1:150, v / v) as the mobile phase to give compound 7 (84 mg, yield 75%) as a green solid. 1 H NMR (400 MHz, CDCl3) δ 8.88 (s, 2H), 7.63(d, J = 6.9 Hz, 4H), 7.49 (t, J = 7.8 Hz, 4H), 7.41 (t, J = 6.0 Hz, 2H),2.80(d, J = 5.9 Hz, 4H), 1.80 – 1.70 (m, 2H), 1.41 – 1.27 (m, 8H), 1.23 (m, 8H),0.89 – 0.79 (m, 12H). 13C NMR (151 MHz, CDCl3) δ 151.31, 144.77, 139.15,136.03, 135.79,134.82, 129.48, 128.53, 127.75, 113.26, 40.56, 32.80, 32.61,28.69, 25.85, 23.09, 14.14, 10.83, 1.03. MALDI-TOF MS Calcd for: C 42 H 47 N4S4 + ([M+H] + ): 736.2708. Found: 736.1809。

[0048] Compound 7 ( Figure 3 a) Characteristic peaks of Raman shift 894 cm -1 and 1264 cm -1 were detected under 785 nm excitation ( Figure 3 b). Under 808 nm excitation with a power density of 1 W / cm 2 , the temperature of compound 7 in 95% water: 5% THF solution was found to increase by 16.4 °C in 10 min ( Figure 3 c).

[0049] The obtained compound 7 has both Raman spectral imaging and photothermal conversion performance.

[0050] Example 4: Synthesis reaction formula of Raman imaging probe 4,7-bis(4-(2-hexyldecyl)thiophen-2-yl)benzo[l,2-c:4,5-c']bis[l,2,5]thiadiazole (compound 9): Reagents and conditions a: tetraphenylphosphine palladium, 1,4-dioxane, 105 °C; was synthesized as follows: (1) Compound 1 (0.3 g, 0.85 mmol) and tetraphenylphosphine palladium (0.15 g, 0.14 mmol) were placed in a 50 mL two-neck flask containing 1,4-dioxane (20 mL), and tributyl(4-(2-hexyldecyl)thiophen-2-yl)stannane (compound 8, 2.05 g, 2.84 mmol) was added under nitrogen protection, and nitrogen was replaced (x3); (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, the organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain the blue solid product compound 9 (0.47 g, 69%). 1 H NMR (400 MHz, CDCl3)δ 8.88 (s, 2H), 7.27 (s, 2H), 2.78 (d, J = 6.9 Hz, 4H), 1.83 – 1.72(m, 2H),1.23 (m, 48H), 0.85 (t, J = 6.4 Hz, 12H). Compound 9 ( Figure 4 a) Characteristic peaks of Raman shift 894 cm -1 and 1264 cm -1 were detected under 785 nm excitation ( Figure 4 b). Under 808 nm excitation with a power density of 1 W / cm 2 , the temperature of compound 9 in a 95% water:5% THF mixed solution was found to increase by 14.1 ℃ within 10 min ( Figure 4 c).

[0051] The obtained compound 9 has both Raman spectral imaging and photothermal conversion performance.

[0052] Example 5: Synthesis reaction formula of Raman imaging probe 4,7-bis((4-(2-hexyldecyl)-5-(4-phenyl)thiophene-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 11): In step a, the reaction reagent and conditions a are: NBS, chloroform, DMF, ice bath; was synthesized by the following steps: (1) NBS (0.14 g, 0.78 mmol) was added dropwise to a chloroform / DMF (1:1, v / v) mixed solution of compound 9 (0.22 g, 0.38 mmol) at room temperature; (2) The mixture was stirred in an ice bath for 10 min under nitrogen protection, then quenched with ice water, and the aqueous phase was extracted with dichloromethane; The organic phase was dried over anhydrous sodium sulfate. After removing dichloromethane under reduced pressure, the residue was purified by silica gel chromatography with ethyl acetate / petroleum ether (1:120, v / v) as eluent to obtain the desired product compound 10 (106 mg, yield 38%) as a blue solid.

[0053] 1 H NMR (600 MHz, CDCl3) δ 8.39 (s, 2H), 2.59 (d, J = 7.2 Hz, 4H), 1.81(d, J = 5.2 Hz, 2H), 1.40 – 1.22 (m, 48H), 0.92 – 0.84 (m, 12H). MALDI-TOF MSCalcd for: C 46 H 69 Br2N4S4 + ([M+H] + ): 964.2672. Found: 964.1844. Step b, the reaction reagent and condition b are as follows: phenylboronic acid, tetrakis(triphenylphosphine)palladium, 2M K2CO3, 1,4-dioxane, 105 °C; Synthesis was carried out as follows: (1) Compound 10 (28 mg, 0.038 mmol), phenylboronic acid (12 mg, 0.10 mmol) and tetrakis(triphenylphosphine)palladium (4 mg, 0.004 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (10 mL) was added by syringe under nitrogen and the resulting solution was degassed by vacuum; (3) 2M K2CO3 solution (0.06 mL) was added under nitrogen; (4) The mixture was further degassed by vacuum and then stirred at 105 °C for 4 h; (5) After removal of the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate; The product was purified by silica gel chromatography using ethyl acetate / petroleum ether (1:150, v / v) as the mobile phase to give compound 11 (24 mg, yield 75%) as a green solid.

[0054] 1 H NMR (400 MHz, CDCl3) δ 8.88 (s, 2H), 7.61 (d, J = 7.1 Hz, 4H), 7.46(t, J = 7.3 Hz, 4H), 7.38 (t, J = 7.1 Hz, 2H), 2.78 (d, J =6.9 Hz, 4H), 1.83 –1.72 (m, 2H), 1.23 (d, J = 19.8 Hz, 48H), 0.85 (t, J = 6.4 Hz, 12H). MALDI-TOF MSCalcd for: C 58 H 79 N4S4 + ([M+H] + ): 958.5104. Found: 958.1652. Compound 11 ( Figure 5 a) Characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 were detected upon excitation at 785 nm ( Figure 5 b). Upon excitation at 808 nm with a power density of 1 W / cm 2 , the temperature of the solution of compound 11 in 95% water: 5% THF was found to increase by 20.5 °C in 10 min ( Figure 5 c).

[0055] The resulting compound 11 exhibited both Raman spectral imaging and photothermal conversion properties.

[0056] Example 6: Preparation and evaluation of a micellar formulation containing compound 11 Preparation of a micellar formulation containing compound 11: One mg of distearoylphosphatidyl ethanolamine-polyethylene glycol 2000 and 0.5 mg of compound 11 Raman imaging probe were dissolved in THF, then added to water and diluted ten-fold to a final volume of 8 mL of mixed solution; After the mixed solution was sonicated for 2 min, it was placed in a 50 mL Erlenmeyer flask and stirred in a fume hood for 6 h. Finally, the mixed solution was filtered through a 0.22 µm filter membrane to prepare a micellar formulation.

[0057] The relevant parameters of the prepared micellar formulation are shown in Table 1: Compound 11 was prepared into a micelle with a particle size of about 100 nm ( Figure 6 a), which is a nano-formulation. The aqueous solution of the compound 11 micellar formulation (10 µM) exhibited characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 upon excitation at 785 nm ( Figure 6 b). Upon excitation at 808 nm with a power density of 1 W / cm2 It can be seen that the temperature of its aqueous solution increased by 22.1 ℃ within 10 minutes. Figure 6 c).

[0058] A mouse model of orthotopic colon cancer was established using CT26-Luc cell line. After intravenous injection of BBTPPRO micelle (7.5 mg / kg) for 24 h, the skin and peritoneum of the cecum were incised to expose the intestinal segment. Raman spectroscopy imaging was performed on the in situ lesion and metastatic lesions. An in situ lesion measuring 3.20 mm × 2.45 mm and a metastatic lesion measuring approximately 1.83 mm × 2.08 mm were detected. Figure 7 a) Raman scattering spectra of the tumor region showed a distinct 894 cm⁻¹ pattern. −1 Characteristic peaks ( Figure 7 b, Region 2 (tumor foci in situ) and Region 3 (tumor metastases). This characteristic peak was not detected in normal colon tissue. Figure 7 b, Region 1). Tumor tissue samples were taken from the images, and serial sections were subjected to H&E staining and Raman spectroscopy imaging. The results showed that the Raman scattering signal distribution areas of both the in situ and metastatic tumors were co-located with the tumor tissue areas shown by H&E staining. Figure 7 c). Guided by Raman spectroscopy imaging, photothermal therapy was performed on the tumor site using optical fibers. The primary tumor was irradiated with an 808 nm laser for 5 minutes, and the metastatic tumor was irradiated with an 808 nm laser for 8 minutes, at a power of 1 W / cm². 2 The temperature at the tumor site can reach above 51 ℃ and be maintained for 100 s ( Figure 7 d, 7e). Mice treated with Raman imaging-guided photothermal therapy (nanoparticle + laser group) showed no tumor recurrence within 70 days, while all mice in the other three groups died within 35 days. Figure 8 ).

[0059] Example 7: Preparation and evaluation of albumin formulations containing compound 9 Preparation of albumin formulations containing compound 9: Dissolve 5 mg of bovine serum albumin in 8 mL of water, dissolve 0.5 mg of the Raman imaging probe in 0.8 mL of THF, and then add the solutions dropwise to the water while stirring to obtain a mixed solution. The mixture was sonicated for 2 min and then placed in a 50 mL eggplant-shaped flask. It was stirred in a fume hood for 6 h. Finally, the mixture was filtered through a 0.22 µm filter membrane to prepare albumin nanoparticles (BBTPRO NPs).

[0060] The relevant parameters of the prepared albumin nanoparticle formulation are shown in Table 2: Compound 9 was prepared into albumin nanoparticles with a particle size of about 190 nm (a). The aqueous solution of the compound 9 albumin preparation (10 μM) was excited by 785 nm, and characteristic peaks at Raman shifts of 894 cm-1 and 1264 cm-1 were detected (b). Excitation by 808 nm at a power density of 1 W / cm2caused the temperature of the aqueous solution to increase by 15.6 ℃ within 10 min (c). Figure 9 -1 -1 Figure 9 2 -1 -1 Figure 9

[0061] Example 8: Preparation and evaluation of nanoparticle preparations containing compound 11 Preparation of nanoparticle preparations containing compound 11: 5 mg of polyethylene glycol-poly (lactic-co-glycolic acid) block copolymer (PLGA, molecular weight 50000, LA:GA = 50:50, PEG, molecular weight 2000) and 0.5 mg of the Raman imaging probe were dissolved in 1 mL of chloroform reagent; After mixing, 10 mL of water was added dropwise, and the mixed solution was ultrasonicated for 2 min, then placed in a 50 mL erlenmeyer flask, stirred in a fume hood for 6 h, and finally filtered through a 0.22 μm filter membrane to prepare nanoparticles (BBTPPRO NPs).

[0062] The relevant parameters of the prepared nanoparticle preparation are shown in Table 3: Compound 11 was prepared into nanoparticles with a particle size of about 95 nm (a). The aqueous solution of the compound 11 nanoparticles (10 μM) was excited by 785 nm, and characteristic peaks at Raman shifts of 894 cm-1 and 1264 cm-1 were detected (b). Excitation by 808 nm at a power density of 1 W / cm2caused the temperature of the aqueous solution to increase by 23.1 ℃ within 10 min (c). Figure 10 -1 -1 Figure 10 2 Figure 10

[0063] Example 9: Synthesis of Raman imaging probe compound 13, reaction scheme: Reaction reagents and conditions a: tetrakis (triphenylphosphine) palladium, 1,4-dioxane, 105 ℃; Synthesized according to the following procedure:​​​​​​​​​​ (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis triphenylphosphine palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 12 (2.04 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x3); (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, and the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain solid product compound 13 (0.36 g, 40%). MALDI-TOF MS Calcd for: C 60 H 53 N6O4S4 + ([M+H] + ): 1048.2933. Found: 1049.3534.

[0064] Compound 13 was detected at 894 cm -1 and 1264 cm -1 Raman shift under 830 nm excitation. Under 808 nm excitation with a power density of 1 W / cm 2 , the temperature of the mixture solution of 95% water:5% THF was found to increase by 20.1 °C within 10 min.

[0065] Example 10: Preparation and evaluation of micellar formulation containing compound 13 Preparation of micellar formulation containing compound 13: 1 mg of distearoylphosphatidyl ethanolamine-polyethylene glycol 5000 and 0.5 mg of the Raman imaging probe were dissolved in 0.8 mL of THF, then added to water and diluted ten times to a final volume of 8 mL of mixed solution; After the mixed solution was ultrasonicated for 2 min, it was placed in a 50 mL flask and stirred in a fume hood for 6 h, and finally the mixed solution was filtered through a 0.22 µm filter membrane to prepare micelles.

[0066] The relevant parameters of the prepared nano-formulation are shown in Table 4: The compound 13 was prepared into micelles with a particle size of about 102 nm. The aqueous solution of the compound 13 micelles (10 μM) was excited at 830 nm, and characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 were detected. When excited at 808 nm with a power density of 1 W / cm 2 , it was found that the heating rate of the compound 13 in the aqueous solution was much higher than that of water.

[0067] A mouse orthotopic colon cancer model was established using the CT26-Luc cell line, and the nanoparticles (20 mg / kg) were injected intravenously 24 h later. The skin and peritoneum at the cecum site were incised to expose the intestinal segment, and Raman spectral imaging was performed on the orthotopic site and metastatic site, respectively. The orthotopic site and metastatic site of the tumor were detected. Under the guidance of Raman spectral imaging, optical fiber was used to implement photothermal therapy on the tumor site. The orthotopic tumor was irradiated with 808 nm laser for 5 minutes, and the metastatic tumor was irradiated with 808 nm laser for 8 minutes. The power was 1 W / cm 2 , and the temperature at the tumor site could reach above 51 ℃ and maintain for 100 s. Within 30 days after treatment, no tumor recurrence was found in the tumor-bearing mice, while the mice that were not guided by Raman imaging for photothermal therapy all relapsed and died within 30 days.

[0068] Example 11: Synthesis of the Raman imaging probe compound 15: Reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; was synthesized according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-neck flask containing 1,4-dioxane (20 mL), and compound 14 (2.24 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x3); (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography using ethyl acetate / petroleum ether (1:20, v / v) as the eluent to obtain solid product compound 15 (0.41 g, 41%). MALDI-TOF MS Calcd for: C 78 H 67 N4S4 + [M+H] +): 1186.4170. Found: 1187.6540.

[0069] Compound 15 can be detected at 894 cm -1 and 1264 cm -1 Raman shift characteristic peaks. When excited by 808 nm laser with a power density of 1 W / cm 2 , it can be seen that the temperature of its 95% water: 5% THF mixed solution increased by 28.3 ℃ within 10 min.

[0070] Example 12: Preparation and evaluation of albumin preparation containing compound 15 Preparation of albumin preparation containing compound 15: (1) 5 mg of human serum albumin was dissolved in 8 mL of water, and 0.5 mg of the Raman imaging probe was dissolved in 0.8 mL of THF, then added dropwise into water under stirring; (2) After the mixed solution was ultrasonicated for 2 min, it was placed in a 50 mL round-bottom flask and stirred in a fume hood for 6 h, and finally the mixed solution was filtered through a 0.22 µm filter membrane to prepare albumin nanoparticles.

[0071] The relevant parameters of the prepared nanopreparation are shown in Table 5: Compound 15 was prepared into albumin nanoparticles with a particle size of about 180 nm. The aqueous solution of the compound 15 albumin preparation (10 µM) can be detected at 894 cm -1 and 1264 cm -1 Raman shift characteristic peaks when excited by 830 nm laser. When excited by 808 nm laser with a power density of 1 W / cm 2 , it can be seen that the temperature of its aqueous solution increased by 27.4 ℃ within 10 min.

[0072] A mouse orthotopic colon cancer model was established using CT26-Luc cell strain, and the nanoparticles (20 mg / kg) were injected intravenously 24 h later. The skin and peritoneum of the cecum part were cut open to expose the intestinal segment, and Raman spectral imaging was performed on the orthotopic lesions and metastatic lesions, respectively, to detect the orthotopic lesions and metastatic lesions of the tumor. Under the guidance of Raman spectral imaging, optical fiber was used to perform photothermal therapy on the tumor site, and 808 nm laser was used to irradiate the orthotopic tumor for 5 min and the metastatic tumor for 8 min, with a power of 1 W / cm 2, the temperature at the tumor site can reach above 51 ℃ and maintain for 100 s. Within 30 days after the treatment, no tumor recurrence was found in the tumor-bearing mice, while in the mice without the Raman image-guided photothermal treatment, all the mice relapsed and died within 30 days.

[0073] Example 13: Synthesis of Raman image probe compound 17 Reagents and conditions a: tetrakis triphenylphosphine palladium, 1,4-dioxane, 105 ℃; Synthesis was carried out according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis triphenylphosphine palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-neck flask containing 1,4-dioxane (20 mL), and compound 16 (1.91 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x 3) to obtain a mixture; (2) The mixture was stirred at 105 ℃ for 12 h, and then cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, and the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain solid product compound 17 (0.24 g, 30%). MALDI-TOF MS Calcd for: C 52 H 83 N4O4S4 + ([M+H] + ): 954.5219. Found: 955.4920.

[0074] Compound 17 can be detected at 894 cm -1 and 1264 cm -1 Raman shift under 785 nm excitation. Under 808 nm excitation with a power density of 1 W / cm 2 , the temperature of the 95% water:5% THF mixed solution can be seen to increase by 15.3 ℃ within 10 min.

[0075] Example 14: Preparation and evaluation of nanoparticle preparation containing compound 17 Preparation of nanoparticle preparation containing compound 17: (1) 2 mg of polyethylene glycol-poly(lactic-co-glycolic acid) block copolymer (PLGA, molecular weight 20000, LA:GA=50:50, PEG, molecular weight 2000) and 0.5 mg of the Raman imaging probe were dissolved in 1 mL of chloroform reagent; (2) After mixing evenly, add to 10 mL of water, and after ultrasonic treatment for 2 min, place the mixed solution in a 50 mL conical flask, stir in a fume hood for 6 h, and finally filter the mixed solution through a 0.22 μm filter membrane to prepare nanoparticles.

[0076] The relevant parameters of the prepared nano-preparation are shown in Table 6: Compound 17 was prepared into nanoparticles with a particle size of about 88 nm. The aqueous solution of the compound 17 nanoparticles (10 μM) can detect characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 under 785 nm excitation. Under 808 nm excitation with a power density of 1 W / cm 2 , the temperature of the aqueous solution was found to increase by 14.1 ℃ within 10 min.

[0077] A mouse orthotopic colon cancer model was established using the CT26-Luc cell line, and the nanoparticles (20 mg / kg) were injected intravenously 24 h later. The skin and peritoneum of the cecum were cut open to expose the intestinal segment, and Raman spectral imaging was performed on the orthotopic lesions and metastatic lesions. The orthotopic lesions and metastatic lesions can be detected. Under the guidance of Raman spectral imaging, optical fiber was used to perform photothermal therapy on the tumor site. The orthotopic tumor was irradiated with 808 nm laser for 5 min, and the metastatic tumor was irradiated with 808 nm laser for 8 min, with a power of 1 W / cm 2 . The temperature of the tumor site can reach more than 51 ℃ and maintain for 100 s. Within 30 days after treatment, no tumor recurrence was found in the tumor-bearing mice, while the mice that did not undergo Raman imaging-guided photothermal therapy all relapsed and died within 30 days.

[0078] Example 15: Synthesis of Raman imaging probe compound 19 Reaction reagents and conditions a: tetra-triphenylphosphine palladium, 1,4-dioxane, 105 ℃; Synthesized according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 18 (1.33 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x3); (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, and the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain solid product compound 19 (0.16 g, 35%). MALDI-TOF MS Calcd for: C 24 H 15 N8S4 + ([M+H] + ): 542.0224. Found: 542.6720.

[0079] Compound 19 exhibited characteristic peaks at 894 cm -1 and 1264 cm -1 Raman shifts upon excitation at 785 nm. Upon excitation at 808 nm with a power density of 1 W / cm 2 , it was observed that the temperature of a 95% water:5% THF mixture solution increased by 24.1 °C within 10 min.

[0080] Example 16: Synthesis of Raman image probe compound 21 Reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; Synthesis was carried out as follows: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 20 (1.26 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x3) to obtain a mixture; (2) After the mixture was stirred at 105 °C for 12 h, it was cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1 :20, v / v) as eluent to give compound 21 (0.17 g, 40%) as a solid. MALDI-TOF MS Calcd for: C 22 H 15 N4O2S4 + ([M+H] + ): 494.0000. Found: 494.6200.

[0081] Compound 21 showed characteristic peaks at 894 cm -1 and 1264 cm -1 Raman shifts upon excitation at 785 nm. Upon excitation at 808 nm with a power density of 1 W / cm 2 , it was observed that the temperature of a 95% water: 5% THF mixture solution increased by 18.6 °C within 10 min.

[0082] Example 17: Synthesis of Raman image probe compound 23 Reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; was synthesized according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 22 (1.54 g, 2.85 mmol) was added under nitrogen protection, and the mixture was replaced with nitrogen (x 3); (2) The mixture was stirred at 105 °C for 12 h, and then cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1 :20, v / v) as eluent to give compound 23 (0.27 g, 45%) as a solid. MALDI-TOF MS Calcd for: C 32 H 31 N4O2S6 + ([M+H] + ): 694.0693. Found: 694.9780.

[0083] Compound 23 can be detected at 894 cm -1 and 1264 cm -1 Raman shift characteristic peaks. Upon excitation at 808 nm with a power density of 1 W / cm 2 It can be seen that the temperature of the 95% water: 5% THF mixture solution increased by 16.7 °C within 10 min.

[0084] Example 18: Synthesis of Raman image probe compound 25 Reagents and conditions a: tetrakis triphenylphosphine palladium, 1,4-dioxane, 105 °C; Synthesis was carried out as follows: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis triphenylphosphine palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 24 (1.30 g, 2.85 mmol) was added under nitrogen protection, and nitrogen was replaced (x3) to obtain a mixture; (2) The mixture was stirred at 105 °C for 12 h, and then cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, and the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain solid product compound 25 (0.19 g, yield 42%). MALDI-TOF MS Calcd for: C 22 H 15 N4O4S4 + ([M+H] + ): 525.9898. Found: 526.6180.

[0085] Compound 23 can be detected at 894 cm -1 and 1264 cm -1 Raman shift characteristic peaks. Upon excitation at 808 nm with a power density of 1 W / cm 2 It can be seen that the temperature of the 95% water: 5% THF mixture solution increased by 16.7 °C within 10 min.

[0086] Example 19: Synthesis of Raman image probe compound 27 Reaction reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; Synthesized according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 26 (1.28 g, 2.85 mmol) was added under nitrogen protection, and the mixture was replaced with nitrogen (x 3); (2) The mixture was stirred at 105 °C for 12 h, and then cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, and the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to obtain solid product compound 27 (0.15 g, 34%). MALDI-TOF MS Calcd for: C 18 H 17 N6O4S4 + ([M+H] + ): 508.0116. Found: 508.6040.

[0087] Compound 27 exhibited characteristic peaks at 894 cm -1 and 1264 cm -1 Raman shifts upon excitation at 785 nm. Upon excitation at 808 nm with a power density of 1 W / cm 2 , it was observed that the temperature of a 95% water:5% THF mixture solution increased by 17.5 °C within 10 min.

[0088] Example 20: Synthesis of Raman image probe compound 29 Reaction reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; Synthesized according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), and compound 26 (1.28 g, 2.85 mmol) was added under nitrogen protection, and the mixture was replaced with nitrogen (x 3); (2) The mixture was stirred at 105 °C for 12 h, and then cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1 :20, v / v) as eluent to give the solid product compound 29 (0.14 g, 36%). MALDI-TOF MS Calcd for: C 16 H5F2N4O2S4 + ([M+H] + ): 449.9185. Found: 450.4708.

[0089] Compound 29 showed characteristic peaks at 894 cm -1 and 1264 cm -1 Raman shifts upon excitation at 785 nm. Upon excitation at 808 nm with a power density of 1 W / cm 2 , a temperature increase of 12.8 °C within 10 min was observed for a solution of compound 29 in 95% water:5% THF.

[0090] Example 21: Synthesis of Raman image probe compound 31: Reagents and conditions a: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105 °C; was synthesized according to the following procedure: (1) Compound 1 (0.3 g, 0.85 mmol) and tetrakis(triphenylphosphine)palladium (98 mg, 0.08 mmol) were placed in a 50 mL two-necked flask containing 1,4-dioxane (20 mL), compound 30 (1.97 g, 2.85 mmol) was added under nitrogen protection, and the mixture was replaced with nitrogen (x 3); (2) The mixture was stirred at 105 °C for 12 h, and then cooled to room temperature; (3) The mixture was poured into water and extracted with dichloromethane, the organic layer was washed with saturated aqueous potassium fluoride solution and brine, then dried over anhydrous sodium sulfate, the solvent was removed by rotary evaporation, and the residue was purified by column chromatography with ethyl acetate / petroleum ether (1 :20, v / v) as eluent to give the solid product compound 31 (0.39 g, 46%). MALDI-TOF MS Calcd for: C 52 H 81 Cl2N4O2S4 + ([M+H] + ): 990.4541. Found: 992.3780.

[0091] Compound 31 showed characteristic peaks of Raman shift at 894 cm -1 and 1264 cm -1 -1 upon excitation at 808 nm with a power density of 1 W / cm 2 -1. It was observed that the temperature of the solution in 95% water: 5% THF increased by 14.9 °C within 10 min.

[0092] Example 22: Synthesis of Raman image probe compound 34 Reaction reagent and condition b: tetrakis triphenylphosphine palladium, 2 M K2C03, 1,4-dioxane, 105 °C; was synthesized according to the following procedure: (1) Compound 32 (250 mg, 0.38 mmol), compound 33 (172 mg, 1.00 mmol) and tetrakis triphenylphosphine palladium (40 mg, 0.04 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (20 mL) was added under nitrogen using a syringe and the resulting solution was degassed by vacuum, to give a mixture; (3) 2 M K2C03solution (0.6 mL) was added under nitrogen; (4) The mixture was further degassed by vacuum and then stirred at 105 °C for 4 h; (5) After removal of the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using ethyl acetate / petroleum ether (1:150, v / v) as the mobile phase to give compound 34 as a solid (207 mg, yield 70%). MALDI-TOF MS Calcd for: C 46 H 43 N4S4 + ([M+H] + ): 778.2292. Found: 779.1100.

[0093] Compound 34 showed characteristic peaks of Raman shift at 894 cm -1 and 1264 cm -1 -1 upon excitation at 808 nm with a power density of 1 W / cm 2It can be seen that the temperature of its aqueous solution increased by 17.9 °C within 10 min.

[0094] Example 23: Preparation and evaluation of the micellar formulation containing compound 34 Preparation of the micellar formulation containing compound 34: (1) 1 mg of distearoylphosphatidyl ethanolamine-polyethylene glycol 800 and 0.5 mg of the Raman imaging probe were dissolved in 0.8 mL of THF, then added to water and diluted ten times to a final volume of 8 mL; (2) After the mixed solution was ultrasonicated for 2 min, it was placed in a 50 mL round-bottom flask and stirred in a fume hood for 6 h. Finally, the mixed solution was filtered through a 0.22 μm filter membrane to prepare the micelles.

[0095] The relevant parameters of the prepared nano-formulation are shown in Table 7: Compound 34 was prepared into micelles with a particle size of about 90 nm. The aqueous solution of the compound 34 micelles (10 μM) could detect characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 under 830 nm excitation. Under 808 nm excitation with a power density of 1 W / cm 2 , it can be seen that the temperature of its aqueous solution increased by 17.7 °C within 10 min.

[0096] A mouse orthotopic colon cancer model was established using the CT26-Luc cell line. After intravenous injection of the formulation (20 mg / kg) for 24 h, the skin and peritoneum of the cecum part were cut open to expose the intestinal segment, and Raman spectral imaging was performed on the orthotopic lesions and metastatic lesions, respectively. The orthotopic lesions and metastatic lesions could be detected. Under the guidance of Raman spectral imaging, optical fiber was used to perform photothermal therapy on the tumor site. The orthotopic tumor was irradiated with 808 nm laser for 5 min, and the metastatic tumor was irradiated with 808 nm laser for 8 min, with a power of 1 W / cm 2 . The temperature of the tumor site could reach more than 51 °C and maintain for 100 s. Within 30 days after treatment, no tumor recurrence was found in the tumor-bearing mice, while the mice that did not undergo Raman imaging-guided photothermal therapy all relapsed and died within 30 days.

[0097] Example 24: Synthesis of Raman imaging probe compound 36 Reaction reagents and conditions b: tetra-triphenylphosphine palladium, 2M K2CO3, 1,4-dioxane, 105 °C; Synthesis was carried out according to the following steps: (1) Compound 32 (250 mg, 0.38 mmol), compound 35 (160 mg, 1.00 mmol) and tetrakis triphenylphosphine palladium (40 mg, 0.04 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (20 mL) was added by syringe under nitrogen and the resulting solution was degassed by vacuum; (3) 2 M K2CO3 solution (0.6 mL) was added under nitrogen; (4) The mixture was further degassed by vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using ethyl acetate / petroleum ether (1:150, v / v) as the mobile phase to give solid compound 36 (172 mg, yield 60%). MALDI-TOF MS Calcd for: C 42 H 41 N6S4 + ([M+H] + ): 756.2197. Found: 757.0640.

[0098] Compound 36 showed characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 when excited at 830 nm. When excited at 808 nm with a power density of 1 W / cm 2 , it was observed that the temperature of a 95% water:5% THF mixture increased by 19.3 °C within 10 min.

[0099] Example 25: Preparation and evaluation of nanoparticles containing compound 36 Preparation of nanoparticles containing compound 36: (1) 5 mg of polyethylene glycol-poly(lactic-co-glycolic acid) block copolymer (molecular weight of PLGA 40000, LA:GA = 40:60, molecular weight of PEG 5000) and 0.5 mg of the Raman imaging probe were dissolved in 1 mL of chloroform reagent; (2) After mixing well, it was added dropwise into 10 mL of water, and the mixed solution was ultrasonicated for 2 min and then placed in a 50 mL tomato-shaped flask, stirred in a fume hood for 6 h, and finally the mixed solution was filtered through a 0.22 µm filter membrane to prepare nanoparticles.

[0100] The relevant parameters of the prepared nano-preparation are shown in Table 8: The compound 36 was prepared into nanoparticles with a particle size of about 98 nm. The aqueous solution of the nanoparticles (10 μM) of the compound 36 had characteristic peaks of Raman shift at 894 cm -1 and 1264 cm -1 , which could be detected under 830 nm excitation. The temperature of the aqueous solution thereof was increased by 19.0 ℃ within 10 min under 808 nm excitation with a power density of 1 W / cm 2 .

[0101] The CT26-Luc cell strain was used to establish a mouse orthotopic colon cancer model, and the nanoparticles (20 mg / kg) were injected intravenously 24 h later. The skin and peritoneum of the cecum part were cut open to expose the intestinal segment, and the Raman spectrum image of the orthotopic lesion and the metastatic lesion was performed, respectively, so that the orthotopic lesion and the metastatic lesion could be detected. Under the guidance of the Raman spectrum image, the optical fiber was used to perform the photothermal treatment on the tumor site, and the orthotopic tumor was irradiated with 808 nm laser for 5 min, and the metastatic tumor was irradiated with 808 nm laser for 8 min, with a power of 1 W / cm 2 , and the temperature of the tumor site could reach above 51 ℃ and maintain for 100 s. Within 30 days after the treatment, no tumor recurrence was found in the tumor-bearing mice, while the mice that were not treated with the Raman image guidance photothermal treatment all relapsed and died within 30 days.

[0102] Example 26: Synthesis of the Raman image probe compound 38: The reaction reagent and condition b are: tetra-triphenylphosphine palladium, 2M K2CO3, 1,4-dioxane, 105 ℃; Synthesis was performed according to the following steps: (1) Compound 32 (250 mg, 0.38 mmol), compound 37 (198 mg, 1.00 mmol) and tetra-triphenylphosphine palladium (40 mg, 0.04 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (20 mL) was added under nitrogen by a syringe, and the resulting solution was degassed by vacuum to obtain a mixture; (3) 2M K2CO3 solution (0.6 mL) was added under nitrogen; (4) The mixture was further degassed by vacuum, and then stirred at 105 ℃ for 4 h; (5) After the solvent was removed under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using ethyl acetate / petroleum ether (1 : 150, v / v) as the mobile phase to give solid compound 38 (252 mg, yield 80%). MALDI-TOF MS Calcd for: C 46 H 39 N8S4 + ([M+H] + ): 830.2102. Found: 831.1060.

[0103] Compound 38 exhibited characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 when excited at 830 nm. When excited at 808 nm with a power density of 1 W / cm 2 , the temperature of its aqueous solution in 95% water: 5% THF increased by 18.0 °C within 10 min.

[0104] Example 27: Preparation and evaluation of albumin formulation containing compound 38 Preparation of albumin formulation containing compound 38: (1) 5 mg of recombinant human serum albumin was dissolved in 8 mL of water, and 0.5 mg of the Raman imaging probe was dissolved in 0.8 mL of THF, which was then added dropwise into water under stirring; (2) After the mixed solution was ultrasonicated for 2 min, it was placed in a 50 mL conical flask and stirred in a fume hood for 6 h. Finally, the mixed solution was filtered through a 0.22 pm filter membrane to prepare albumin nanoparticles.

[0105] The relevant parameters of the prepared nanopreparation are shown in Table 9: Compound 38 was prepared into albumin nanoparticles with a particle size of about 150 nm. The aqueous solution of the compound 38 albumin formulation (10 pM) exhibited characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 when excited at 830 nm. When excited at 808 nm with a power density of 1 W / cm 2 , the temperature of its aqueous solution increased by 17.8 °C within 10 min.

[0106] CT26-Luc cell line was used to establish orthotopic colon cancer model in mice. After 24 h of intravenous injection of the nanoparticles (20 mg / kg), the skin and peritoneum of the cecum were cut open to expose the intestinal segment, and Raman spectral imaging was performed on the orthotopic and metastatic lesions. The orthotopic and metastatic lesions were detected. Under the guidance of Raman spectral imaging, optical fiber was used to perform photothermal therapy on the tumor site. The orthotopic tumor was irradiated with 808 nm laser for 5 min, and the metastatic tumor was irradiated with 808 nm laser for 8 min, with a power of 1 W / cm 2 . The temperature of the tumor site could reach above 51 ℃ and maintain for 100 s. Within 30 days after treatment, no tumor recurrence was found in the tumor-bearing mice, while the mice that did not undergo Raman imaging-guided photothermal therapy all relapsed and died within 30 days.

[0107] Example 28: Synthesis of Raman imaging probe compound 40: Reaction reagent and condition b: tetrakis triphenylphosphine palladium, 2 M K2CO3, 1,4-dioxane, 105 ℃; was synthesized according to the following procedure: (1) Compound 32 (250 mg, 0.38 mmol), compound 39 (239 mg, 1.00 mmol), and tetrakis triphenylphosphine palladium (40 mg, 0.04 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (20 mL) was added by syringe under nitrogen, and the resulting solution was degassed by vacuum; (3) 2 M K2CO3 solution (0.6 mL) was added under nitrogen; (4) The mixture was further degassed by vacuum, and then stirred at 105 ℃ for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography with ethyl acetate / petroleum ether (1:150, v / v) as the mobile phase to obtain solid compound 40 (260 mg, yield 75%). MALDI-TOF MS Calcd for: C 54 H 53 N6S4 + ([M+H] + ): 912.3136. Found: 913.2920.

[0108] Compound 40 can be detected by 830 nm excitation with Raman shift 894 cm-1 and 1264 cm -1 -1. Upon excitation at 808 nm with a power density of 1 W / cm 2 -1, the temperature of its aqueous solution was observed to increase by 24.0 °C within 10 min.

[0109] Example 29: Preparation and evaluation of a micellar formulation containing compound 40 Preparation of a micellar formulation containing compound 40: (1) 1 mg of distearoylphosphatidyl ethanolamine-polyethylene glycol 2000 and 0.5 mg of the Raman imaging probe were dissolved in 800 μL of THF, and then added to water and diluted ten-fold to a final volume of 8 mL; (2) After the mixed solution was ultrasonicated for 2 min, it was placed in a 50 mL erlenmeyer flask, stirred in a fume hood for 6 h, and finally the mixed solution was filtered through a 0.22 μm filter membrane to prepare the micelles.

[0110] The relevant parameters of the prepared nano-formulation are shown in Table 10: Compound 40 was prepared into micelles with a particle size of about 110 nm. The aqueous solution of the compound 40 micelles (10 μM) could detect characteristic peaks at Raman shifts of 894 cm -1 and 1264 cm -1 -1 upon excitation at 808 nm with a power density of 1 W / cm 2 -1. The temperature of its aqueous solution was observed to increase by 24.0 °C within 10 min.

[0111] A mouse orthotopic colon cancer model was established using the CT26-Luc cell line. After intravenous injection of the formulation (20 mg / kg) for 24 h, the skin and peritoneum of the cecum were cut open to expose the intestinal segment, and Raman spectral imaging was performed on the orthotopic lesions and metastatic lesions. The orthotopic lesions and metastatic lesions could be detected. Under the guidance of Raman spectral imaging, optical fiber was used to perform photothermal therapy on the tumor site. The orthotopic tumor was irradiated with 808 nm laser for 5 min, and the metastatic tumor was irradiated with 808 nm laser for 8 min, with a power of 1 W / cm 2 -1. The temperature of the tumor site could reach above 51 °C and maintain for 100 s. Within 30 days after treatment, no tumor recurrence was found in the tumor-bearing mice, while the mice that were not treated with Raman imaging-guided photothermal therapy all relapsed and died within 30 days.

[0112] Example 30: Synthesis of Raman imaging probe compound 42 Reaction reagents and conditions b: tetrakis triphenylphosphine palladium, 2 M K2CO3, 1,4-dioxane, 105 °C; Prepared according to the following procedure: (1) Compound 32 (250 mg, 0.38 mmol), compound 41 (434 mg, 1.00 mmol) and tetrakis triphenylphosphine palladium (40 mg, 0.04 mmol) were placed in a 50 mL two-necked flask; (2) Degassed 1,4-dioxane (20 mL) was added via syringe under nitrogen and the resulting solution was degassed by vacuum; (3) 2 M K2CO3 solution (0.6 mL) was added under nitrogen; (4) The mixture was further degassed by vacuum and then stirred at 105 °C for 4 h; (5) After removal of the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried over anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using ethyl acetate / petroleum ether (1:150, v / v) as the mobile phase to give compound 42 as a solid (347 mg, 70% yield). MALDI-TOF MS Calcd for: C 84 H 111 N4S4 + ([M+H] + ): 1304.0720. Found: 1304.2027.

[0113] Compound 42 showed characteristic Raman shifts at 894 cm -1 and 1264 cm -1 when excited at 830 nm. When excited at 808 nm with a power density of 1 W / cm 2 , it was observed that the temperature of a 95% water:5% THF mixture solution increased by 25.3 °C within 10 min.

Claims

1. A Raman imaging probe with photothermal conversion capability, having an 894 cm⁻¹ diameter. -1 and 1264 cm -1 The Raman shift characteristic peaks, the molecular structure of the Raman imaging probe is based on thiophene-coupled benzobisthiadiazole as the parent core, and its general structural formula I is as follows: General Formula I In I, R1, R2, R3, and R4 are -H, -F, -Cl, -Br, -I, -OH, -CHO, -NH2, -COOH, and -C, respectively. n H 2n+1 , -OC n H 2n+1 , -(0CH2) n CH3, substituted or unsubstituted Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Any one of them.

2. A method for preparing a Raman imaging probe according to claim 1, characterized in that, include: Synthesis Route 1: The target product was obtained by Stille reaction using 4,7-dibromobenzobisthiadiazole and α- or β-substituted tributyl(thiophene-2-yl)tinane as starting materials, and the reaction formula (1) is as follows: （1） Among them, R1, R2, R3, and R4 are -H, -F, -Cl, -Br, -I, -OH, -CHO, -NH2, -COOH, and -C, respectively. n H 2n+1 , -OC n H 2n+1 , -(0CH2) n CH3, substituted or unsubstituted Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution any of them, In reaction formula (1), reagent a is tetrakis(triphenylphosphine)palladium and 1,4-dioxane, and the reaction temperature is 105 °C; or, Synthesis Route 2: Using the β-substituted 4,7-dithienylbenzobisthiadiazole obtained from synthetic route 1 as the starting material, the target product was obtained by bromination and Suzuki reaction, as shown in reaction formula (2) below: （2） Among them, R1, R2, R3, and R4 are -H, -F, -Cl, -Br, -I, -OH, -CHO, -NH2, -COOH, and -C, respectively. n H 2n+1 , -OC n H 2n+1 , -(0CH2) n CH3, substituted or unsubstituted Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution Substitution or non-substitution any of them, In the reaction formula (2), The reagents used were: N-bromosuccinimide (NBS), chloroform, and N,N-dimethylformamide (DMF), and the reaction conditions were an ice bath. Reagent b is 2 M K2CO3, tetra-triphenylphosphine palladium, aryl, heteroarylboronic acid or aryl, heteroarylpinacol boronic acid ester, 1,4-dioxane, and the reaction temperature is 105 °C.

3. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for the Raman imaging probe 4,7-bis(4-(2-propyl)thiophen-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 3) is as follows: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. The steps are as follows: (1) 0.6 g, 1.70 mmol of 4,7-dibromo-benzo[1,2-c:4,5-c']bis([1,2,5]thiadiazole (compound 1) and 196 mg, 0.16 mmol of tetratriphenylphosphine palladium were placed in a 25 mL 50 mL double-necked flask containing 1,4-dioxane. Under nitrogen protection, 2.36 g, 5.68 mmol of tributyl((2-propyl)thiophen-2-yl)stanane (compound 2) was added, and the mixture was purged with nitrogen three times. (2) After stirring the mixture at 105°C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with dichloromethane / petroleum ether (1:1, v / v) as eluent to give 376 mg of compound 3 in 50% yield as a blue solid.

4. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for the Raman imaging probe 4,7-bis(4-(2-ethylhexyl)thiophen-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 5, BBT) is as follows: The reaction reagents and conditions a are: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105℃; The steps are as follows: (1) Place 0.3 g, 0.85 mmol of 4,7-dibromo-benzo[1,2-c:4,5-c']bis([1,2,5]thiadiazole (compound 1) and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane. Under nitrogen protection, add 1.38 g, 2.85 mmol of tributyl(4-(2-ethylhexyl)thiophen-2-yl)stanane (compound 4). Replace with nitrogen three times. (2) After stirring the mixture at 105°C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to give 0.27 g of compound 5 in 56% yield, which was a blue solid product BBT.

5. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for the Raman imaging probe 4,7-bis((4-(2-ethylhexyl)-5-(4-phenyl)thiophen-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 7) is as follows: In step a, Reagents and conditions: (a) N-bromosuccinimide (NBS), chloroform, N,N-dimethylformamide (DMF), ice bath; Follow these steps: (1) At room temperature, 0.22 g, 0.38 mmol of NBS (0.14 g, 0.78 mmol) was added dropwise to a chloroform / DMF (10 mL, 1:1, v / v) mixed solution of BBT; (2) Stir the mixture in an ice bath for 10 min under nitrogen protection, then quench it with ice water, and extract the aqueous phase with dichloromethane; (3) Wash with water and saturated brine, and dry the organic phase with anhydrous sodium sulfate; (4) After removing dichloromethane under reduced pressure, the residue was purified by silica gel chromatography and eluted with ethyl acetate / petroleum ether at a volume ratio (v / v) of 1:120 to obtain the desired product compound 6 in a blue solid with a yield of 38% (106 mg). In step b, Reagents and conditions: (b) phenylboronic acid, tetrakis(triphenylphosphine)palladium, 2M K₂CO₃, 1,4-dioxane, 105 °C; proceed as follows: (1) Place 112 mg, 0.15 mmol of compound 6, 48 mg, 0.38 mmol of phenylboronic acid and 16 mg, 0.02 mmol of tetraphenylphosphine palladium in a 50 mL double-necked flask; (2) Add 20 mL of degassed 1,4-dioxane to nitrogen using a syringe, and degassed the resulting solution under vacuum to obtain a mixture; (3) Add 0.6 mL of 2 M K2CO3 solution under a nitrogen atmosphere; (4) The mixture was further degassed under vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:150 (v / v) as the mobile phase to obtain 84 mg of green solid compound 7 in 75% yield.

6. The method for preparing the Raman imaging probe according to claim 2, characterized in that, Synthesis of Raman imaging probe 4,7-bis(4-(2-hexyldecyl)thiophen-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 9): The reaction reagents and conditions a were tetrakis(triphenylphosphine)palladium, 1,4-dioxane, and 105 °C. Follow these steps: (1) Place 0.3 g and 0.85 mmol of 4,7-dibromo-benzo[1,2-c:4,5-c']bis([1,2,5]thiadiazole (compound 1) and 0.15 g and 0.14 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane. Under nitrogen protection, add 2.05 g and 2.84 mmol of tributyl(4-(2-hexyldecyl)thiophen-2-yl)stanane (compound 8), and purge with nitrogen three times. (2) After stirring the mixture at 105°C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with ethyl acetate / petroleum ether as eluent at a volume ratio (v / v) of 1:20 to give 0.47 g of compound 9 with a yield of 69%, which was a blue solid product.

7. The method for preparing the Raman imaging probe according to claim 2, characterized in that, Synthesis of Raman imaging probe 4,7-bis((4-(2-hexyldecyl)-5-(4-phenyl)thiophene-2-yl)benzo[1,2-c:4,5-c']bis[1,2,5]thiadiazole (compound 11): In step a, Reagents and conditions: (a) NBS, chloroform, DMF, ice bath; Follow these steps: (1) At room temperature, 0.22 g, 0.38 mmol of NBS (0.14 g, 0.78 mmol) was added dropwise to a chloroform / DMF mixture of compound 9 in a volume ratio (v / v) of 1:

1. (2) The mixture was stirred in an ice bath for 10 min under nitrogen protection, then quenched with ice water, and the aqueous phase was extracted with dichloromethane. The mixture was washed with water and saturated brine, and the organic phase was dried over anhydrous sodium sulfate. (3) After removing dichloromethane under reduced pressure, the residue was purified by silica gel chromatography and eluted with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:120 (v / v) to obtain the desired 106 mg of blue solid compound 10 in 38% yield. In step b, Reagents and conditions: (b) phenylboronic acid, tetrakis(triphenylphosphine)palladium, 2M K₂CO₃, 1,4-dioxane, 105 °C; proceed as follows: (1) Place 28 mg of compound 10 (0.038 mmol), 12 mg of phenylboronic acid (0.10 mmol), and 4 mg of tetraphenylphosphine palladium (0.004 mmol) in a 50 mL double-necked flask; (2) Add 10 mL of degassed 1,4-dioxane to nitrogen using a syringe, and degassed the resulting solution under vacuum; (3) Add 0.06 mL of 2 M K2CO3 solution under nitrogen atmosphere to obtain a mixture; (4) After further vacuum degassing the mixture, stir at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane to obtain an organic phase; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:150 (v / v) as the mobile phase to obtain 24 mg of green solid compound 11 in 75% yield.

8. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 13 is as follows: The reaction reagents and conditions a are: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105℃; Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, and add 2.04 g, 2.85 mmol of compound 12 under nitrogen protection. Replace with nitrogen three times. (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.36 g of solid product compound 13 in 40% yield.

9. The method for preparing the Raman imaging probe according to claim 2, characterized in that, Synthesis reaction formula of Raman imaging probe compound 15: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, and add 2.24 g, 2.85 mmol of compound 14 under nitrogen protection. Replace with nitrogen three times. (2) After stirring the mixture at 105°C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.41 g of solid product compound 15 in a yield of 41%.

10. The method for preparing the Raman imaging probe according to claim 2, characterized in that, Synthesis reaction formula of Raman imaging probe compound 17: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, add 1.91 g, 2.85 mmol of compound 16 under nitrogen protection, and replace with nitrogen three times to obtain a mixture; (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.24 g of solid product compound 17 in 30% yield.

11. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 19 is as follows: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, and add 1.33 g, 2.85 mmol of compound 18 under nitrogen protection. Replace with nitrogen three times. (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.16 g of solid product compound 19 in a yield of 35%.

12. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 21 is as follows: The reaction reagents and conditions a are: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105℃; Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, add 1.26 g, 2.85 mmol of compound 20 under nitrogen protection, and replace with nitrogen three times to obtain a mixture; (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.17 g of solid product compound 21 in 40% yield.

13. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 23 is as follows: The reaction reagents and conditions a are: tetrakis(triphenylphosphine)palladium, 1,4-dioxane, 105℃; Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, add 1.54 g, 2.85 mmol of compound 22 under nitrogen protection, and replace with nitrogen three times to obtain a mixture; (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.27 g of solid product compound 23 in 45% yield.

14. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 25 is as follows: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium were placed in a 50 mL double-necked flask containing 1,4-dioxane in 20 mL. 1.30 g, 2.85 mmol of compound 24 was added under nitrogen protection. The mixture was purged with nitrogen three times to obtain a mixture. (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.19 g of solid product compound 25 in a yield of 42%.

15. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 27 is as follows: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium were placed in a 50 mL double-necked flask containing 1,4-dioxane in 20 mL. 1.28 g, 2.85 mmol of compound 26 was added under nitrogen protection. The mixture was purged with nitrogen three times to obtain a mixture. (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with ethyl acetate / petroleum ether (1:20, v / v) as eluent to give 0.15 g of solid product compound 27 in 34% yield.

16. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 29 is as follows: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) Place 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium in a 20 mL 50 mL double-necked flask containing 1,4-dioxane, add 1.20 g, 2.85 mmol of compound 28 under nitrogen protection, and replace with nitrogen three times to obtain a mixture; (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give solid product compound 29 (0.14 g, 36%).

17. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 31 is as follows: The reaction reagents and conditions a were: tetratetraphenylphosphine palladium, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) 0.3 g, 0.85 mmol of compound 1 and 98 mg, 0.08 mmol of tetratriphenylphosphine palladium were placed in a 20 mL 50 mL double-necked flask containing 1,4-dioxane. 1.97 g, 2.85 mmol of compound 30 was added under nitrogen protection. The mixture was purged with nitrogen three times to obtain a mixture. (2) After stirring the mixture at 105 °C for 12 h, cool it to room temperature; (3) The mixture was poured into water and extracted with dichloromethane. The organic layer was washed with saturated potassium fluoride aqueous solution and brine, then dried with anhydrous sodium sulfate. The solvent was removed by rotary evaporation. The residue was purified by column chromatography with a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:20 (v / v) as the eluent to give 0.39 g of solid product compound 31 in a yield of 46%.

18. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 34 is as follows: The reaction reagents and conditions b are: tetrakis(triphenylphosphine)palladium, 2M K2CO3, 1,4-dioxane, 105℃; Synthesize using the following steps: (1) Place 250 mg, 0.38 mmol of compound 32, 172 mg, 1.00 mmol of compound 33 and 40 mg, 0.04 mmol of tetraphenylphosphine palladium in a 50 mL double-necked flask; (2) Add 20 mL of degassed 1,4-dioxane to nitrogen using a syringe, and degassed the resulting solution under vacuum to obtain a mixture; (3) Add 0.6 mL of 2 M K2CO3 solution under nitrogen atmosphere; (4) The mixture was further degassed under vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:150 (v / v) as the mobile phase to obtain 207 mg of solid compound 34 in 70% yield.

19. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 36 is as follows: The reaction reagents and conditions b were: tetratetraphenylphosphine palladium, 2M K2CO3, 1,4-dioxane, and 105 °C. Synthesize according to the following steps. (1) Place 250 mg, 0.38 mmol of compound 32; 160 mg, 1.00 mmol of compound 35 and 40 mg, 0.04 mmol of tetraphenylphosphine palladium in a 50 mL double-necked flask; (2) Add 20 mL of degassed 1,4-dioxane using a syringe under nitrogen atmosphere, and degassed the resulting solution under vacuum. (3) Add 0.6 mL of 2 M K2CO3 solution under nitrogen atmosphere; (4) The mixture was further degassed under vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:150 (v / v) as the mobile phase to obtain 172 mg of solid compound 36 in 60% yield.

20. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 38 is as follows: The reaction reagents and conditions b were: tetratetraphenylphosphine palladium, 2M K2CO3, 1,4-dioxane, and 105 °C. Synthesize using the following steps: (1) Place 250 mg, 0.38 mmol of compound 32; 198 mg, 1.00 mmol of compound 37 and 40 mg, 0.04 mmol of tetraphenylphosphine palladium in a 50 mL double-necked flask; (2) Add 20 mL of degassed 1,4-dioxane using a syringe under nitrogen atmosphere, and degassed the resulting solution under vacuum to obtain a mixture; (3) Add 0.6 mL of 2 M K2CO3 solution under nitrogen atmosphere; (4) The mixture was further degassed under vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio (v / v) of 1:150 as the mobile phase to give 252 mg of solid compound 38 in 80% yield.

21. The method for preparing the Raman imaging probe according to claim 2, characterized in that, Synthesis of Raman imaging probe compound 40: Reagents and conditions b: tetrakis(triphenylphosphine)palladium, 2M K₂CO₃, 1,4-dioxane, 105 °C; Synthesize using the following steps: (1) Place 250 mg, 0.38 mmol of compound 32; 239 mg, 1.00 mmol of compound 39; and 40 mg, 0.04 mmol of tetratriphenylphosphine palladium in a 50 mL double-necked flask; (2) Add 20 mL of degassed 1,4-dioxane using a syringe under nitrogen atmosphere, and degassed the resulting solution under vacuum. (3) Add 0.6 mL of 2 M K2CO3 solution under nitrogen atmosphere; (4) The mixture was further degassed under vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:150 (v / v) as the mobile phase to obtain 260 mg of solid compound 40 in 75% yield.

22. The method for preparing the Raman imaging probe according to claim 2, characterized in that, The synthetic reaction formula for Raman imaging probe compound 42 is as follows: The reaction reagents and conditions b were: tetrakis(triphenylphosphine)palladium, 2M K2CO3, 1,4-dioxane, and 105 °C. Prepare according to the following steps: (1) Place 250 mg, 0.38 mmol of compound 32; 434 mg, 1.00 mmol of compound 41; and 40 mg, 0.04 mmol of tetratriphenylphosphine palladium in a 50 mL double-necked flask; (2) Add 20 mL of degassed 1,4-dioxane using a syringe under nitrogen atmosphere, and degassed the resulting solution under vacuum. (3) Add 0.6 mL of 2 M K2CO3 solution under nitrogen atmosphere; (4) The mixture was further degassed under vacuum and then stirred at 105 °C for 4 h; (5) After removing the solvent under reduced pressure, the resulting residue was extracted with dichloromethane; (6) The organic phase was washed with water and saturated brine and dried with anhydrous sodium sulfate; (7) The product was purified by silica gel chromatography using a mixed solvent of ethyl acetate / petroleum ether at a volume ratio of 1:150 (v / v) as the mobile phase to obtain 347 mg of solid compound 42 in 70% yield.

23. The use of a Raman imaging probe with photothermal conversion capability according to claim 1, characterized in that, This formulation is used to prepare Raman imaging probes with photothermal conversion capabilities. In the aggregated state, it does not require a metal substrate for enhancement. When irradiated with 700-900 nm near-infrared light, it generates a strong Raman scattering signal and can convert light energy into heat energy. It can be used in therapeutic formulations for in vivo Raman spectroscopy imaging and photothermal therapy.

24. A micelle formulation encapsulating the Raman imaging probe of claim 1, characterized in that, The Raman imaging probe is encapsulated in a micelle material to form a micelle formulation, the composition of which includes the Raman imaging probe and distearylphosphatidylethanolamine-polyethylene glycol and its derivatives. The mass ratio of the Raman imaging probe to distearylphosphatidylethanolamine-polyethylene glycol and its derivatives is 1:0.2-1:50, and the micelle particle size is 50-1000 nm. The distearylphosphatidylethanolamine-polyethylene glycol and its derivatives are selected from one or a mixture of several of the following: distearylphosphatidylethanolamine-polyethylene glycol, distearylphosphatidylacetamide-polyethylene glycol-amino, distearylphosphatidylacetamide-polyethylene glycol-maleimide, and distearylphosphatidylacetamide-polyethylene glycol-mercapto.

25. The micelle formulation according to claim 24, characterized in that, The mass ratio of the Raman imaging probe to distearate phosphatidylethanolamine-polyethylene glycol and its derivatives is 1:0.2-1:

10.

26. The micelle formulation according to claim 24 or 25, characterized in that, The molecular weight of polyethylene glycol is between 500 and 5000, and the distearylphosphatidylethanolamine-polyethylene glycol and its derivatives are distearylphosphatidylacetamide-polyethylene glycol 2000.

27. A method for preparing a micelle formulation according to any one of claims 24 to 26, comprising the following steps: (1) Distearate phosphatidylethanolamine-polyethylene glycol 2000 and the Raman imaging probe are dissolved in chloroform, dichloromethane, tetrahydrofuran or a mixture thereof in a mass ratio to obtain a mixed solution; (2) Add the mixed solution described in step (1) dropwise to water and dilute it tenfold to obtain a diluted solution; (3) The diluted solution obtained in (2) is sonicated for 2-10 min, stirred for 8-12 h, and then filtered through a 0.22 µm filter membrane to prepare micelles.

28. An albumin nanoformulation encapsulating the Raman imaging probe of claim 1, characterized in that, The Raman imaging probe is encapsulated in albumin to prepare an albumin formulation, the composition of which includes the Raman imaging probe and albumin, wherein the mass ratio of the Raman imaging probe to albumin is 1:0.2-1:50, and the particle size of the albumin nanoparticles is 50-1000 nm; wherein the albumin is selected from one or a mixture of several of human serum albumin, recombinant human serum albumin, or bovine serum albumin; the Raman imaging probe aggregates in the formulation and, upon irradiation with near-infrared light, emits light at 894 cm⁻¹. -1 and 1264cm -1 Raman shift generates strong Raman scattering signals for in vivo Raman spectral imaging, and can also convert light energy into heat energy as a photothermal therapy drug for tumors.

29. The nano-formulation according to claim 28, characterized in that, The mass ratio of the Raman imaging probe to albumin is 1:0.2-1:

10.

30. A method for preparing a nano-formulation according to claim 28 or 29, characterized in that, The steps include the following: (1) Dissolve albumin in water to obtain an aqueous albumin solution with a concentration of 0.1%-5% (w / v); (2) Dissolve the Raman imaging probe in methanol, ethanol, propylene glycol, chloroform, dichloromethane, tetrahydrofuran or a mixture thereof, wherein the ratio of the organic solvent to the water in step (1) is 1:40-1:60; (3) Add the organic solution from step (2) to the albumin aqueous solution from step (1) according to the mass ratio of the Raman imaging probe to albumin and mix them evenly to obtain a mixture; (4) Stir the mixture obtained in step (3) at room temperature for 1-8 h.

31. The method for preparing nano-formulations according to claim 30, characterized in that, In step (1), the concentration of albumin aqueous solution is 0.5%-2% (w / v).

32. A nanoparticle formulation encapsulating the Raman imaging probe of claim 1, characterized in that, A nanoformation was prepared by encapsulating the Raman imaging probe in a polyethylene glycol block copolymer material. The nanoformation comprises the Raman imaging probe and the polyethylene glycol block copolymer, with a mass ratio of Raman imaging probe to polyethylene glycol block copolymer of 1:0.2-1:

50. The nanoparticle size is 50-1000 nm. The polyethylene glycol block copolymer is selected from one or more of polyethylene glycol-poly(lactic-co-glycolic acid) block copolymer, polyethylene glycol-polycaprolactone block copolymer, polyethylene glycol-polylactic acid block copolymer, and polyethylene glycol-polycarbonate block copolymer. The molecular weight of the polyethylene glycol block copolymer ranges from 2000 to 60000, and the molecular weight of polyethylene glycol (PEG) ranges from 1000 to 5000. The Raman imaging probe aggregates in the formulation and, upon irradiation with near-infrared light, emits light at 894 cm⁻¹. -1 and 1264 cm -1 Raman shift generates strong Raman scattering signals for in vivo Raman spectral imaging, and can also convert light energy into heat energy as a photothermal therapy drug for tumors.

33. The nanoparticle formulation of the Raman imaging probe according to claim 32, characterized in that, The mass ratio of the Raman imaging probe to the polyethylene glycol block copolymer is 1:0.2-1:

10.

34. The nanoparticle formulation of the Raman imaging probe according to claim 32 or 33, characterized in that, The polyethylene glycol block copolymer is polyethylene glycol-polylactic acid-glycolic acid block copolymer (PEG-PLGA), with PLGA having a molecular weight of 50,000, lactic acid (LA) to glycolic acid (GA) ratio of LA:GA = 50:50, and PEG having a molecular weight of 2,000.

35. The method for preparing the nanoparticle formulation of the Raman imaging probe according to claim 34, characterized in that, The PEG-PLGA nanoformulation containing the Raman imaging probe comprises the following preparation steps: (1) Dissolve the Raman imaging probe and PEG-PLGA in methanol, ethanol, propylene glycol, chloroform, dichloromethane, tetrahydrofuran or a mixture thereof in a mass ratio to obtain an organic solution; (2) Under ultrasonic and ice-water bath conditions, add the organic solution from step (1) to pure water and mix evenly; (3) Stir the mixture obtained in step (2) at room temperature for 1-8 h to evaporate the organic solvent and obtain a nanoparticle solution; (4) Freeze-dry the nanoparticle solution obtained in step (3) to obtain the desired nanoparticle formulation.

36. The method for preparing the nanoparticle formulation of the Raman imaging probe according to claim 35, characterized in that, The mass ratio of the Raman imaging probe to PEG-PLGA in step (1) is 1:0.2-1:

10.

37. The method for preparing the nanoparticle formulation of the Raman imaging probe according to claim 35, characterized in that, The ratio of organic solvent to pure water in step (2) is 1:10-1:

20.

38. A method of using the formulation according to any one of claims 24 to 26, 28 or 29, and 32 or 33, characterized in that, In vivo Raman spectroscopy imaging can be performed without the need for surface enhancement of a metal substrate when excited by lasers with wavelengths of 785 nm and 830 nm; when excited by lasers with wavelength of 808 nm, light energy can be converted into heat energy for tumor hyperthermia.

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