Hemicyanine dye with integrated light diagnosis and treatment performance

Abstract

The invention relates to a compound shown in a formula (I) and a compound shown in a formula (II), which can be used as a hemicyanine dye with integrated optical diagnosis and treatment performance, and application of the hemicyanine dye in tumor optical diagnosis and treatment, especially fluorescence and photoacoustic imaging.

Description

Technical Field

[0001] The present invention relates to a semi-cyanine dye and its application as a photosensitive molecule with controllable photodegradation, especially in the field of tumor photodiagnosis and phototherapy, belonging to the technical field of medical materials. Background Art

[0002] Photodiagnosis and phototherapy integration is an emerging field that focuses on developing theranostic agents that can serve as both diagnostic tools and therapeutic agents; these agents typically use light, especially near-infrared (NIR) radiation, as the energy source for imaging and treatment to form high-quality imaging, enabling these agents to have great potential in diagnosing and treating various diseases. In the past decade, due to the reduced photon scattering in biological tissues resulting in less background signal interference, fluorescence imaging techniques in the second near-infrared (NIR-II, 1000 - 1700 nm) region have been used to provide high-contrast imaging and sensing in vivo. Although NIR-II fluorescence imaging has improved the penetration depth of fluorescence imaging techniques, difficulties are still encountered when imaging deeper lesions. An effective way to overcome the limitations of single-modal imaging is to combine it with other imaging modalities. Photoacoustic imaging is the most convenient method to achieve complementary advantages with near-infrared fluorescence imaging because both technologies rely on light excitation; in addition, photoacoustic imaging can penetrate deeper than near-infrared fluorescence, but it has lower sensitivity to soft tissues. Phototherapy, especially photodynamic therapy (PDT) and photothermal therapy (PTT), has received extensive attention due to its non-contact nature, convenience, and low risk. Through NIR-II fluorescence and photoacoustic imaging-guided phototherapy, precise diagnosis and treatment of tumors can be achieved simultaneously, enabling real-time evaluation of the treatment effect. PDT mainly involves type I and type II mechanisms. Compared with the high oxygen dependence of type II photodynamics, the type I process is an ideal candidate for hypoxic cancer treatment, which can enhance the PDT response under hypoxic conditions. In addition, PTT can improve the treatment effect because it not only treats tumors through temperature elevation during the treatment process but also provides additional oxygen for hypoxic tumor tissues by promoting blood supply between tumors. Therefore, an ideal phototherapy method should exhibit strong NIR-II fluorescence / photoacoustic signals for imaging and have excellent type I PDT / PTT performance.

[0003] To date, most phototheranostic agents rely on the repurposing of existing fluorophores. Among them, cyanine dyes have been widely used in early studies. Due to the easy synthesis of hemicyanine dyes (HDs), they have attracted considerable attention in this field. HDs with a D-π-A structure have several advantages, including high molar absorption coefficients, large Stokes shifts, good biocompatibility, easy molecular modification, and adjustable absorption and fluorescence wavelengths. Although some near-infrared derivatives of hemicyanine have been designed and synthesized by modifying electron donor and acceptor groups, few of them simultaneously exhibit the multiple properties required for multimodal diagnosis or treatment and are biodegradable. Therefore, the development of hemicyanine dyes with "all-in-one" phototheranostic properties remains a major challenge. Summary of the Invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide a hemicyanine dye with a D-π-A structure, which can form J-aggregates through self-assembly. The nanoparticles of this hemicyanine dye can not only exhibit excellent NIR-II fluorescence and photoacoustic imaging properties, but also exhibit photodynamic and photothermal characteristics, making it an "all-in-one" phototherapeutic agent. In addition, the nanoparticles of this hemicyanine dye can be oxidized by HClO, which is beneficial to its biodegradation during the macrophage process and reduces the metabolic burden. At the same time, its nanoparticles also have good targeting effects on tumors and can clearly locate tumors through NIR-II fluorescence imaging and photoacoustic imaging. The study on the effect of phototherapy on subcutaneous tumor models shows that the hemicyanine dye nanoparticles of the present invention can effectively ablate tumors with low toxicity and side effects.

[0005] Specifically, the technical solution of the present invention is as follows:

[0006] On the one hand, the present invention provides a compound represented by formula (I) and formula (ID) with a D-π-A structure:

[0007]

[0008] Wherein,

[0009] Ring D, D1, and D2 are each an electron donor, and each is an optionally C 1-12 alkoxy, amine-substituted C6-C 12 aryl or 5-16 membered heteroaryl, and the heteroaryl contains at least one N atom;

[0010] Ring A is an electron acceptor; it is an optionally C 1-6 alkyl, C 1-6 haloalkyl, 5-7 membered heteroaryl or C 6-12 aryl-substituted tricyanofuran-based electron acceptor.

[0011] In a specific embodiment of the present invention, ring D, D1, and D2 are each independently selected from:

[0012]

[0013] wherein,

[0014] R1 and R2 are each independently C 1-12 alkyl;

[0015] R3 is each independently C 1-12 alkyl or benzyl, and the alkyl is optionally further substituted by -OSi(Ph)2-C(CH3)3.

[0016] In a specific embodiment of the present invention, ring A is

[0017] wherein, R4 and R5 are each independently C 1-6 alkyl, C 1-6 haloalkyl, 5-7 membered heteroaryl or C 6-12 aryl, and the heteroaryl contains at least one heteroatom selected from O, N, and S. Preferably, R4 and R5 are each independently C 1-4 alkyl, C 1-4 haloalkyl, 5-7 membered heteroaryl or C 6-7 aryl.

[0018] In a preferred embodiment of the present invention, ring A is:

[0019] More preferably, ring A is

[0020] In a preferred embodiment of the present invention, ring D, D1, and D2 are each independently selected from any one of the following structures:

[0021]

[0022] In an embodiment of the present invention, the compounds of formula (I) and formula (II) are selected from the following compounds:

[0023]

[0024] On the other hand, the present invention provides a method for preparing a compound having the structure shown in formula (I) or formula (II).

[0025] The compounds of formula (I) and formula (II) of the present invention can be prepared by the following synthetic route. Specifically, the compounds of formula (I) and formula (II) are synthesized from an aldehyde compound of an electron donor D and a formyl chloride compound containing electron donors D1 and D2 to obtain the compounds of formula (I) and formula (II).

[0026] 1. Preparation of the compound of formula (I)

[0027]

[0028] First, an aldehyde compound of an aniline-based electron donor and isophorone mixed solution react under the condition of sodium alkoxide to obtain intermediate compound 1. Second, compound 1 reacts in an acetonitrile solution containing n-butyllithium to obtain intermediate compound 2, which reacts with diisobutylaluminum hydride to obtain intermediate compound 3; intermediate compound 3 reacts with diethyl cyanomethylphosphonate under alkaline conditions to obtain intermediate compound 4. Third, the aforementioned synthesis is repeated to obtain intermediate compound 5. Finally, intermediate compound 5 reacts with a cyano-furan-based electron acceptor A under alkaline conditions to obtain the compound of formula (I).

[0029] 2. Preparation of the compound of formula (II)

[0030]

[0031] First, a formyl chloride compound of electron donor D2 reacts with electron donor D1 compound to obtain intermediate compound 1 of formula (II). Second, intermediate compound 1 reacts in the presence of CH3Li to obtain intermediate compound 2. Finally, intermediate compound 2 reacts with intermediate compound 3 under alkaline conditions to obtain the compound of formula (ID).

[0032] On the other hand, the present invention provides a hemicyanine dye, and the dye is a compound of formula (I) or formula (II) having a D-π-A structure.

[0033] On the other hand, the present invention provides the application of the compound having the structure shown in formula (I) or formula (II) in hemicyanine dyes.

[0034] On the other hand, the present invention also provides the application of the compound having the structure shown in formula (I) or formula (II) in fluorescence and photoacoustic imaging.

[0035] On the other hand, the present invention also provides the application of the compound having the structure shown in formula (I) or formula (II) in integrated photodiagnosis and treatment, especially as an integrated photodiagnosis and treatment preparation.

[0036] The compounds of formula (I) and formula (II) of the present invention can be used as semi-cyanine dyes with a D-π-A structure and can form J-aggregates through self-assembly. The maximum absorption of the nanoparticles of this dye is near 808 nm, and the absorption spectrum can be extended to the second near-infrared region (>1000 nm). Therefore, it can be excited by an 808-nm laser or a second near-infrared laser; and it exhibits excellent NIR-II fluorescence and photoacoustic imaging performance after being excited. At the same time, since the maximum emission wavelength of the fluorescence of the nanoparticles is about 1100 nm, it also exhibits photodynamic and photothermal properties, making the compounds of formula (I) and formula (ID) of the present invention an "integrated" phototherapeutic agent. In addition, the nanoparticles of this dye can be oxidized by HClO, which is beneficial to its biodegradation in the process of macrophages and reduces the metabolic burden. The nanoparticles also have good targeting effects on tumors. The fluorescence at the tumor site shows stronger signals than organs such as the liver and kidneys, and tumors can be clearly located through NIR-II fluorescence imaging and photoacoustic imaging. The research on the effect of phototherapy on a subcutaneous tumor model shows that under laser irradiation, the temperature at the tumor site injected with nanoparticles increases significantly, the nanoparticles can effectively ablate tumors, and the toxic and side effects are low.

[0037] The concept, specific structure and technical effects of the present invention will be further described below in conjunction with the drawings to fully understand the purpose, features and effects of the present invention. Description of the Drawings

[0038] Figure 1 Showing the ultraviolet absorption of compound C1064-2 of the present invention, as well as TEM and fluorescence images; a) Ultraviolet absorption diagrams of C1064-2 (10 μM) in various organic solvents (1,4-dioxane, toluene, tetrahydrofuran, dichloromethane, chloroform, ethyl acetate), (b) Absorption spectra of C1064-2 (10 μM) in THF and C1064-2 NPs in water; (c) Size distribution and TEM image of C1064-2 NPs; (d) Fluorescence spectrum of C1064-2 NPs in water and NIR-II fluorescence images of C1064-2 NPs at different concentrations (10 μM, 25 μM, 50 μM, 100 μM) in water.

[0039] Figure 2 Showing the fluorescence intensity (I / I0) of DCHF used for the detection of total ROS by compound C1064-2 of the present invention; (b) APF for the detection of OH, (c) DHR123 for the detection of O2·-; (d) Absorbance at 415 nm in the presence or absence of 10 μM C1064-2NP in water and the change of irradiation time with an 808-nm or 1064-nm laser (1 W / cm 2 )

[0040] Figure 3 The concentration-dependent temperature change of the compound C1064-2(a) of the present invention under irradiation at 808 nm (1 W / cm 2 ); (b) The temperature change of C1064-2 NPs in aqueous solution (100 μM / mL) using five cycles of laser irradiation; (c) The negative natural logarithm plot of the cooling time (10 minutes) and temperature driving force of 100 uM C1064-2 NPs; (d) The PA images of C1064-2 NP solutions with different concentrations. Detailed implementation manners

[0041] The present disclosure will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present disclosure and not to limit the scope of the present disclosure. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are weight percentages and weight parts respectively. The experimental materials and reagents used in the following embodiments can be obtained from commercial channels without special instructions.

[0042] Example 1 Preparation of C1064-1

[0043]

[0044] Step 1: Synthesis of Compound 1

[0045] Under nitrogen protection, Na (0.23 g, 10 mmol) was reacted with 10 mL of absolute ethanol to form sodium alkoxide. A mixture of 4-(diethylamino)-2-(3-(isopropyldiphenylsilyloxy)propoxy)benzaldehyde (3.0 g, 6.1 mmol) and isophorone (1.7 g, 12.2 mmol) dissolved in absolute ethanol was slowly added, and the mixture was stirred overnight at 65 °C. The reaction solution was cooled to room temperature, quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, and purified by column chromatography (ethyl acetate: petroleum ether = 1:5) to obtain 2.7 g of a red oily compound 1 with a yield of 72.58%.

[0046] Step 2: Synthesis of Compound 2

[0047] At -78 °C, 2.5 M n-butyllithium (11.8 mL, 29.4 mmol) was slowly added dropwise to a mixed solution of 10 mL of tetrahydrofuran and acetonitrile (1.5 mL, 29.4 mmol). The reaction was carried out at -78 °C for 30 min. Then, a solution of compound 1 (3 g, 4.9 mmol) dissolved in tetrahydrofuran was slowly added. After reacting at -78 °C for 10 min, the reaction was continued at room temperature for 10 min. A small amount of acetic acid was added, and the mixture was stirred at 70 °C overnight. 20 mL of water was added to the reaction solution to quench the reaction. After extraction, the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (ethyl acetate: petroleum ether = 1:20) to obtain 2.1 g of compound 2 with a yield of 67.74%; LCMS (ESI): m / z = 633.3 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.79 - 7.72 (dd, J = 8, 2 Hz, 1H), 7.71 - 7.64 (m, 4H), 7.51 - 7.30 (m, 7H), 7.10 (d, J = 16.4 Hz, 0.5H), 7.05 (d, J = 16 Hz, 0.5H), 6.80 (d, J = 16.3 Hz, 0.5H), 6.74 (d, J = 16 Hz, 0.5H), 6.65 (s, 0.5H), 6.37 - 6.30 (m, 1H), 6.20 (d, J = 2 Hz, 1H), 6.15 (s, 0.5H), 4.23 (t, J = 6.2 Hz, 2H), 3.94 (t, J = 5.8 Hz, 2H), 3.40 (q, J = 6.9 Hz, 4H), 2.44 (d, J = 1.6 Hz, 1H), 2.22 (s, 2H), 2.20 (d, J = 1.6 Hz, 1H), 2.18 - 2.10 (m, 2H), 1.22 (t, J = 6.9 Hz, 6H), 1.07 (s, 9H), 0.99 (s, 3H), 0.95 (s, 3H).

[0048] Step 3: Synthesis of compound 3

[0049] Compound 2 (2.8 g, 4.4 mmol) and 10 mL of dry toluene were added to a round-bottom flask. At -78 °C, 1 M diisobutylaluminum hydride (8.8 mL, 8.8 mmol) was slowly added dropwise under nitrogen protection. After addition, the mixture was stirred at -78 °C for 2 h. Then, silica gel powder containing 10% water was added, and the stirring was continued for 2 h. The reaction mixture was removed from the ice bath after adding a small amount of anhydrous sodium sulfate and reacted at room temperature for 4 h. The reaction solution was filtered through a sintered glass funnel, and the filtrate was concentrated and purified by column chromatography (ethyl acetate: petroleum ether = 1:5) to obtain 1.9 g of a red solid powder with a yield of 67.86%; LCMS (ESI): m / z = 636.3 [M+H] + . 11H NMR (400 MHz, CDCl3) δ 10.20 (d, J = 8.4 Hz, 0.3H), 10.04 (d, J = 8.8 Hz, 0.7H), 7.74 - 7.62 (m, 4H), 7.10 - 7.14 (m, 7H), 7.13 (s, 0.3H), 7.08 (d, J = 15.6 Hz, 0.3H), 7.08 (d, J = 16.4 Hz, 0.7H), 6.82 (d, J = 16.4 Hz, 0.3H), 6.79 (d, J = 16.4 Hz, 0.7H), 6.32 (m, 1H), 6.22 (s, 0.7H), 6.19 (d, J = 2 Hz, 1H), 5.90 (d, J = 8.8 Hz, 0.7H), 5.69 (d, J = 8.4 Hz, 0.3H), 4.23 (t, J = 6.2 Hz, 2H), 3.99 - 3.89 (m, 2H), 3.41 (q, J = 7.0 Hz, 4H), 2.60 (s, 2H), 2.28 (s, 2H), 2.19 - 2.05 (m, 2H), 1.21 (t, J = 7.0 Hz, 6H), 1.07 (s, 9H), 1.02 (s, 3H), 0.99 (s, 3H).

[0050] Step 4: Synthesis of Compound 4

[0051] Dissolve compound 3 (2.6 g, 4.1 mmol) and NaH (0.1 g, 4.92 mmol) in dry 1,2 - dichloroethane solution, slowly add diethyl cyanomethylphosphonate (0.9 g, 4.92 mmol) dropwise, and react at room temperature overnight. Extract the reaction solution with 20 ml of DCM, dry it over anhydrous sodium sulfate, remove the organic solvent, and then separate by column chromatography (ethyl acetate:petroleum ether = 1:10) to obtain 1.8 g of compound 4 with a yield of 66.67%. 11H NMR (400 MHz, CDCl3) δ 7.74 (dd, J = 7.7, 1.7 Hz, 0.3H), 7.68 (dd, J = 7.9, 1.4 Hz, 4H), 7.57 (dd, J = 12, 15.6 Hz, 0.4H), 7.47 - 7.32 (m, 7.4H), 7.20 (dd, J = 12, 12.4 Hz, 0.2H), 7.05 - 6.95 (m, 1H), 6.84 - 6.73 (m, 0.9H), 6.59 (d, J = 9.2 Hz, 0.4H), 6.50 (d, J = 12.3 Hz, 0.2H), 6.32 (ddd, J = 10.5, 8.1, 3.6 Hz, 1H), 6.27 (d, J = 12.5 Hz, 0.2H), 6.24 (s, 0.2H), 6.21 (s, 0.9H), 6.13 (s, 0.3H), 6.07 (d, J = 11.9 Hz, 0.3H), 5.85 (d, J = 12.0 Hz, 0.3H), 5.20 (dd, J = 15.6, 11.9 Hz, 0.6H), 5.07 - 4.99 (m, 0.4H), 4.28 - 4.17 (m, 2H), 3.94 (t, J = 5.8 Hz, 2H), 3.40 (q, J = 6.9 Hz, 4H), 2.28 (s, 1H), 2.25 (s, 0.5H), 2.22 (s, 2H), 2.18 (s, 1H), 2.15 - 2.09 (m, 2H), 1.24 - 1.18 (m, 6H), 1.07 (s, 9H), 0.98 (s, 3.6H), 0.96 (s, 1.2H), 0.95 (s, 1.2H).

[0052] Step 5: Synthesis of Compound 5

[0053] Add compound 4 (1.2 g, 1.8 mmol) and 5 mL of dry toluene into a round-bottom flask. Slowly add 1 M diisobutylaluminum hydride (3.6 mL, 3.6 mmol) dropwise at -78 °C under nitrogen protection. React at -78 °C for 2 h. Add silica gel powder containing 10% water and continue stirring for 2 h. After adding a small amount of anhydrous sodium sulfate to the reaction mixture, remove it from the ice bath and react at room temperature for 4 h. Filter the reaction mixture through a sintered funnel, concentrate the filtrate, and purify it by column chromatography (ethyl acetate:petroleum ether = 1:5) to obtain 0.7 g of red solid powder with a yield of 58.33%. 11H NMR (400 MHz, CDCl3) δ 9.59 (d, J = 8.4 Hz, 0.6H), 9.58 (d, J = 8.4 Hz, 0.4H), 7.72 (d, J = 12.1 Hz, 0.4H), 7.68 (dd, J = 7.9, 1.4 Hz, 4H), 7.53 (dd, J = 14.7, 12.1 Hz, 0.6H), 7.45 (d, J = 8.9 Hz, 1H), 7.43 - 7.32 (m, 6H), 7.02 (dd, J = 19.0, 16.3 Hz, 1H), 6.82 (dd, J = 16.2, 10.7 Hz, 1H), 6.74 (s, 0.4H), 6.35 - 6.30 (m, 1H), 6.26 (d, J = 12.1 Hz, 0.6H), 6.21 (s, 0.6H), 6.20 (s, 1H), 6.15 (dd, J = 13.5, 6.9 Hz, 0.6H), 6.09 (dd, J = 13.7, 7.0 Hz, 0.4H), 6.05 (d, J = 12.5 Hz, 0.4H), 4.26 - 4.20 (m, 2H), 3.95 (t, J = 5.9 Hz, 2H), 3.45 - 3.35 (m, 4H), 2.38 (s, 1.2H), 2.25 (s, 2H), 2.24 (s, 0.8H), 2.14 (p, J = 6.0 Hz, 2H), 1.22 (t, J = 7.0, 2.4H), 1.21 (t, J = 7.0, 3.6H), 1.07 (s, 9H), 1.00 (s, 3.6H), 0.97 (s, 2.4H).

[0054] Step 6: Preparation of C1064 - 1

[0055] Dissolve compound 5 (50 mg, 0.08 mmol) and 2-(3-cyano-4,5,5-trimethylfuran-2(5H)-ylidene)malononitrile (20 mg, 0.1 mmol) in methanol, add 20 μL of pyridine dropwise, and reflux at 70 °C overnight. Concentrate the reaction solution and separate it by column chromatography (dichloromethane:petroleum ether = 2:1) to obtain 31 mg of a black solid product with a yield of 45.93%. LCMS (ESI): m / z = 843.4 [M + H] + . 11H NMR (400 MHz, CDCl3) δ 7.72 - 7.65 (m, 4H), 7.65 - 7.58 (m, 1H), 7.55 - 7.46 (m, 1H), 7.45 - 7.31 (m, 7H), 7.09 (d, J = 16.1 Hz, 1H), 6.83 (dd, J = 11.6, 7.9 Hz, 1H), 6.45 (dd, J = 13.4, 12.1 Hz, 1H), 6.32 (dd, J = 9.1, 2.1 Hz, 1H), 6.30 (s, 1H), 6.27 (d, J = 5.8 Hz, 2H), 6.19 (d, J = 1.9 Hz, 1H), 4.23 (t, J = 6.2 Hz, 2H), 3.94 (t, J = 5.7 Hz, 2H), 3.42 (q, J = 7.0 Hz, 4H), 2.39 (s, 2H), 2.28 (s, 2H), 2.19 - 2.10 (m, 2H), 1.70 (s, 6H), 1.22 (t, J = 7.0 Hz, 6H), 1.07 (s, 9H), 1.01 (s, 6H). 13 13C NMR (101 MHz, CDCl3) δ 176.16, 172.57, 158.59, 151.14, 149.57, 148.90, 147.16, 144.57, 135.46, 133.69, 129.73, 129.63, 129.19, 128.34, 127.88, 127.70, 127.51, 126.18, 115.07, 113.95, 112.69, 111.87, 111.69, 104.84, 96.61, 95.27, 94.07, 64.69, 60.35, 44.63, 39.72, 39.16, 32.28, 31.09, 28.30, 26.87, 26.49, 19.28, 12.76.

[0056] Preparation of Example 2 C1064 - 2

[0057]

[0058] Step 1: Synthesis of Compound 7

[0059] 4 - Dimethylaminobenzoyl chloride (1.83 g, 10 mmol) and AlCl3 (1.47 g, 11 mmol) were dispersed in dry CH2Cl2, and Compound 6 (3 g, 10 mmol) was slowly added. The mixture was stirred at room temperature for 5 hours. The reaction was quenched by adding water, extracted with CH2Cl2, dried over anhydrous Na2SO4, and purified by column chromatography (dichloromethane / hexane = 1:5) to obtain Compound 7 as a yellow oil (yield: 60%). LCMS (ESI): m / z = 449.3 [M + H] +. 1 1H NMR (400 MHz, CDCl3) δ 7.81 (d, J = 9.0 Hz, 2H), 7.18 (s, 1H), 6.65 (d, J = 9.0 Hz, 2H), 3.66 (t, J = 6.8 Hz, 2H), 3.23 - 3.16 (m, 2H), 3.15 - 3.10 (m, 2H), 3.07 (s, 6H), 1.83 - 1.71 (m, 4H), 1.44 (s, 6H), 1.38 - 1.25 (m, 2H), 1.26 (s, 6H), 1.15 - 1.05 (m, 2H), 0.74 (t, J = 7.4 Hz, 3H). 13 13C NMR (101 MHz, CDCl3) δ 193.93, 156.66, 151.31, 132.52, 128.06, 111.40, 74.50, 47.43, 47.24, 40.69, 40.12, 36.48, 32.82, 32.31, 31.81, 31.42, 29.89, 18.83, 13.90.

[0060] Step 2: Synthesis of Compound 8:

[0061] Under nitrogen protection, 0.9 mL of CH3Li (1.6 M) was slowly added to a solution of Compound 7 (0.6 g, 1.3 mmol) in anhydrous THF at 0 °C. After addition, the mixture was stirred at room temperature for 5 hours. The reaction mixture was poured into water, and the organic layer was extracted with dichloromethane and dried over anhydrous Na2SO4. The organic solvent was removed, and the product was purified by column chromatography (dichloromethane / hexane = 1:10) to obtain Compound 8 as a colorless oil (yield: 90%). LCMS (ESI): m / z = 447.3 [M + H] + . 1 1H NMR (400 MHz, CDCl3) δ 7.330 (d, J = 8.9 Hz, 2H), 6.88 (s, 1H), 6.69 (d, J = 8.9 Hz, 2H), 5.50 (d, J = 1.8 Hz, 1H), 5.20 (d, J = 1.7 Hz, 1H), 3.81 (t, J = 6.8 Hz, 2H), 3.14 - 3.10 (m, 2H), 3.09 - 3.04 (m, 2H), 2.98 (s, 6H), 1.84 - 1.75 (m, 4H), 1.43 (s, 6H), 1.24 (s, 6H), 0.82 (t, J = 7.4 Hz, 3H). 1313C NMR (101 MHz, CDCl3) δ 155.45, 147.71, 128.47, 128.03, 113.08, 112.87, 99.91, 71.59, 47.84, 47.22, 39.44, 39.14, 37.10, 32.76, 32.21, 32.02, 31.92, 30.43, 30.03, 29.65, 29.35, 27.08, 22.68, 19.72, 19.01, 14.14.

[0062] Step 3: Synthesis of C1062-2:

[0063] Dissolve compound 8 (50 mg, 0.11 mmol), 2-((4-(E-2-(E)-5-(tert-butyl)-2-chloro-3-(ethoxymethylene)cyclohex-1-en-1-yl)vinyl)-3-cyano-5,5-dimethylfuran-2(5H)-ylidene)malononitrile (48 mg, 0.11 mmol) and 2 drops of pyridine in 3 mL of chloroform, and reflux for 12 hours. Remove the organic solvent and purify by column chromatography (ethyl acetate / hexane = 1:3) to obtain C1064-2 as a green powder (yield: 70%). LCMS (ESI): m / z = 838.4 [M+H] + . 1 1H NMR (400 MHz, CDCl3) δ 8.07 (d, J = 15.7 Hz, 1H), 7.37 (dd, J = 13.1, 8.7 Hz, 2H), 7.32 (d, J = 13.2 Hz, 1H), 6.89 (dd, J = 13.2, 12.4 Hz, 1H), 6.74 (s, 1H), 6.70 (s, 2H), 6.51 (d, J = 15.7 Hz, 1H), 3.64 (t, J = 9.6 Hz, 2H), 3.30 - 2.90 (m, 10H), 2.72 (d, J = 15.9 Hz, 1H), 2.26 - 2.07 (m, 3H), 1.96 (s, 1H), 1.78 (s, 10H), 1.47 (s, 3H), 1.41 (s, 3H), 1.35 - 1.24 (m, 5H), 1.24 - 1.10 (m, 4H), 1.05 (s, 9H), 0.76 (t, J = 7.4 Hz, 3H). 1313C NMR (101 MHz, CDCl3) δ 175.74, 173.69, 156.42, 151.32, 151.01, 146.37, 144.64, 143.75, 138.57, 138.19, 131.98, 130.49, 130.40, 129.95, 129.80, 129.60, 125.54, 125.36, 122.70, 122.55, 119.17, 119.10, 118.98, 113.13, 112.47, 111.64, 110.82, 96.95, 71.84, 47.56, 47.09, 40.81, 40.24, 37.12, 32.87, 32.41, 32.18, 31.94, 31.80, 30.39, 30.28, 30.26, 29.69, 28.32, 27.34, 27.02, 26.97, 19.09, 19.02, 14.12, 14.10.

[0064] Example 3 Preparation of C1064 - 3

[0065]

[0066] Step 1: Synthesis of Compound 10:

[0067] Compound 10 was synthesized according to the procedure of Compound 7. LCMS (ESI): m / z = 405.3 [M + H] + . 1 1H NMR (400 MHz, CDCl3) δ 7.84 (d, J = 9.0 Hz, 2H), 7.16 (s, 1H), 6.67 (d, J = 9.0 Hz, 2H), 3.53 (s, 3H), 3.22 - 3.17 (m, 2H), 3.16 - 3.11 (m, 2H), 3.08 (s, 6H), 1.83 - 1.72 (m, 4H), 1.43 (s, 6H), 1.25 (s, 6H). 13 13C NMR (101 MHz, CDCl3) δ 194.76, 158.05, 153.01, 132.35, 127.66, 110.57, 61.31, 47.55, 47.02, 40.21, 40.16, 36.76, 32.75, 32.20, 31.37, 30.09.

[0068] Step 2: Synthesis of Compound 11:

[0069] Compound 11 was synthesized according to the procedure of Compound 8. LCMS (ESI): m / z = 407.3 [M + H] + . 11H NMR (400 MHz, CDCl3) δ 7.32 (d, J = 8.4 Hz, 1H), 6.91 (s, 1H), 6.78 (s, 1H), 5.55 (s, 1H), 5.22 (s, 1H), 3.58 (s, 3H), 3.17 - 3.11 (m, 2H), 3.10 - 3.03 (m, 2H), 3.01 (s, 6H), 1.80 (s, 4H), 1.40 (s, 6H), 1.27 (s, 6H). 13 13C NMR (101 MHz, CDCl3) δ 156.43, 147.55, 130.26, 128.30, 128.08, 114.14, 96.84, 59.98, 47.53, 37.11, 32.87, 32.03, 31.92, 31.45, 30.17, 29.71, 29.37.

[0070] Step 3: Synthesis of C1064 - 3:

[0071] Synthesize C1064 - 3 according to the synthesis method of C1064 - 2. LCMS (ESI): m / z = 796.4 [M + H] + . 1 1H NMR (400 MHz, CDCl3) δ 8.06 (d, J = 15.7 Hz, 1H), 7.41 - 7.35 (m, 3H), 6.95 (dd, J = 12.1, 8.3 Hz, 1H), 6.78 (d, J = 8.4 Hz, 1H), 6.69 (d, J = 8.8 Hz, 2H), 6.52 (d, J = 15.8 Hz, 1H), 3.47 (s, 3H), 3.22 - 3.10 (m, 4H), 3.05 (s, 6H), 2.72 (d, J = 14.1 Hz, 1H), 2.25 - 210 (m, 2H), 1.88 - 1.73 (m, 10H), 1.43 (s, 3H), 1.39 (s, 3H), 1.26 - 1.22 (m, 7H), 1.06 (s, 9H). 1313C NMR (101 MHz, CDCl3) δ 175.70, 173.67, 157.65, 151.60, 150.99, 146.18, 144.59, 143.63, 137.37, 132.28, 130.76, 130.64, 129.84, 129.49, 129.47, 129.26, 125.56, 122.52, 119.38, 118.71, 113.35, 112.42, 111.65, 110.78, 96.98, 60.39, 60.02, 47.55, 47.10, 42.59, 42.51, 40.54, 40.20, 37.14, 32.80, 32.44, 32.20, 32.01, 31.78, 30.40, 28.26, 27.80, 27.37, 27.00, 14.20.

[0072] Example 4: Preparation and Performance Testing of Nanoparticles

[0073] (1) Preparation of C1064-1 and C1064-2 Nanoparticles

[0074] Dissolve C1064-1 (0.84 mg) or C1064-2 (0.84 mg) and DSPE-PEG2000 (1 mg) separately in 1 mL of THF. Under ultrasonic treatment (12 W, 25 °C), pour the solution into 10 mL of deionized water. Further ultrasonic treat the resulting dispersion (12 W, 25 °C) for 5 minutes. Then stir the mixture at room temperature for two days to obtain C1064-1 NPs and C1064-2 NPs nanoparticles respectively. Among them, the ultraviolet absorption, TEM images and fluorescence images of C1064-2 NPs nanoparticles are as Figure 1 shown.

[0075] (2) Test for the Ability of Nanoparticles to Generate ROS:

[0076] Use dichlorodihydrofluorescein (DCHF) as a fluorescent probe to detect ROS in the solution. Dissolve 10 μM C1064-2 NPs nanoparticles in 1 mL of water containing 10 μM DCHF. Irradiate the mixture with an 808 nm laser (1 W / cm 2 2), and record the fluorescence change of the sample at 530 nm with a fluorescence spectrophotometer (excitation wavelength: 480 nm), as Figure 2 shown in a.

[0077] (3) Test for the Ability of Nanoparticles to Generate 1O2:

[0078] Use 1,3-diphenylisobenzofuran (DBPF) as a probe to monitor the 1Generation of O₂. A 10 μL methanol (1 mM) solution of DPBF was mixed with an aqueous solution of C1064-2 NPs nanoparticles (10 μM), and irradiated with an 808 nm or 1064 nm laser (1 W / cm 2 ). The absorbance at 415 nm was recorded every 10 seconds, as shown in Figure 2 b.

[0079] (4) Test for the ability of nanoparticles to generate O₂·⁻:

[0080] Dihydro rhodamine 123 (DHR123) was used as a fluorescent probe to detect O₂·⁻ in the solution. 10 μM C1064-2 NPs nanoparticles were dissolved in 1 mL of water containing 60 μM DHR 123. The mixture was irradiated with an 808 nm laser (1 W / cm 2 ), and the fluorescence change of the sample at 530 nm was recorded using a fluorescence spectrophotometer (excitation wavelength: 480 nm), as shown in Figure 2 c.

[0081] (5) Oxidation and degradation performance of C1064-2 nanoparticles:

[0082] The biodegradability of C1064-2 nanoparticles was further evaluated by measuring the change in the absorption spectrum of a C1064-2 nanoparticle solution (10 μM) in the presence of 100 μM HClO. As shown in Figure 2 d, as the reaction time extended, the absorption value gradually decreased, indicating that C1064-2 nanoparticles could be oxidized and degraded by HClO. Therefore, destroying this rigid framework would not only accelerate the clearance from the body but also inactivate the phototherapeutic performance of the corresponding molecule, thereby reducing its phototoxicity.

[0083] (6) Photothermal performance test of C1064-2 nanoparticles

[0084] Solutions of C1064-2 NPs with different concentrations (0, 10, 30, 50, 70, 100, 150 μM) were irradiated with an 808 nm laser (1 W / cm 2 ) or a 1064 nm laser (1 W / cm 2 ). The temperature change within 10 min of irradiation was monitored by an infrared camera, as shown in Figure 3 a. After irradiating a solution with a concentration of 100 μM with an 808 nm laser (1 W / cm 2 ) or a 1064 nm laser (1 W / cm 2 ) for 10 min, the irradiation was stopped for 10 min and then irradiated again, repeating 5 cycles, and the temperature change was monitored, as shown in Figure 3 b. The photothermal conversion efficiency was calculated from the cooling process in Figure 3b, as shown in Figure 3 c.

[0085] (7) Photothermal performance test of C1064-2 nanoparticles

[0086] The photoacoustic signals of C1064-2 nanoparticles with different concentrations (10 μM, 25 μM, 50 μM, 100 μM) under the excitation of 808 nm laser or 1064 nm laser were collected by the photoacoustic imaging system TOMO LOIS-3D, as Figure 3 shown in d.

[0087] Example 5

[0088] BALB / c nude mice at 6-7 weeks old were provided by the Experimental Animal Center of North Sichuan Medical College. Under sterile conditions, 1×10 6 4T1 cells were subcutaneously injected into the right anterior axilla of nude mice, and then they were individually housed under specific pathogen-free conditions, with free access to food and water until the formed tumors grew to a diameter of about 1 cm measured by calipers; the growth process took about one week.

[0089] (1) Test of C1064-2 nanoparticles for in vivo near-infrared second-window fluorescence imaging

[0090] Using a small animal NIR-II fluorescence imaging system, the NIR-II fluorescence signals of the whole body of 4T1 tumor-bearing mice were collected at different time points after tail vein injection with 808 nm or 1064 nm excitation. The experimental results showed that almost no NIR-II fluorescence signal was detected in the nude mice before injecting the C1064-2 NPs solution. Once 100 μL of C1064-2 NP (100 μM) solution was injected through the tail vein, obvious NIR-II fluorescence appeared throughout the nude mice. One hour after injection, the NIR-II fluorescence signal at the tumor site was clearly observed. However, as time extended (after 20 h), the fluorescence of the whole nude mouse body irradiated with 808 nm laser became stronger, making it difficult to distinguish the tumor from other sites. By shortening the exposure time from 100 ms to 30 ms, the overall fluorescence intensity could be reduced. The results showed that the fluorescence intensity at the tumor site was significantly higher than that at other sites. This result indicated that C1064-2 NPs could target the tumor site for NIR-II fluorescence imaging and were mainly metabolized through the hepatobiliary system.

[0091] (2) Test of C1064-2 nanoparticles for in vivo photoacoustic imaging:

[0092] Similar to the second near-infrared imaging, in vivo photoacoustic imaging analysis was also performed by collecting photoacoustic signals at different time points before injection and after intravenous injection of 100 μL of C1064-2NP (100 μM) in 4T1 tumor-bearing mice. The experimental results showed that after injection of C1064-2 NPs, the photoacoustic signals gradually increased, and the tumor site could be clearly observed after 30 hours. Moreover, the cross-section of the tumor site also showed obvious asymmetric photoacoustic signals, indicating the location of the tumor. This result indicates that C1064-2 NP can actively target the tumor site for photoacoustic imaging.

[0093] (3) Characterization of the anti-tumor performance of C1064-2 nanoparticles for phototherapy

[0094] The 4T1 tumor-bearing mice were randomly divided into 3 groups, named "C1064-2 NPs", "C1064-2 NPs + 808 nm laser", and "C1064-2 NPs + 1064 nm laser", respectively. 100 μL of C1064-2 NPs (200 μM) was intratumorally injected into the tumor sites of the mice, and after injection, the mice were irradiated with 808 nm and 1064 nm lasers (1 W / cm 2 ) for 20 min, respectively. During the laser irradiation, infrared thermal images of the mice were obtained using an infrared camera, and the tumor volume and body weight were measured every day for 14 consecutive days. Ex vivo tumor photos and tumor H&E-stained sections of the mice 14 days after treatment showed that the tumors on the tumor-bearing mice were significantly reduced and gradually disappeared, indicating that C1064-2 nanoparticles have obvious photoinhibitory effects on tumors under laser irradiation.

Claims

1. A compound of the structure represented by formula (I) or formula (II): in, Ring D, D1, D2 are each optionally selected from: Ring A is in, R1 and R2 are C 1-12 alkyl; R3 are respectively C 1-12 alkyl or benzyl, and the alkyl is optionally further substituted with -OSi(Ph)2-C(CH3)3; R4 and R5 are C 1-6 Alkyl, C 1-6 haloalkyl, 5-7 membered heteroaryl or C 6-12 wherein the heteroaryl group contains at least one heteroatom selected from O, N, and S.

2. The compound according to claim 1, wherein R4 and R5 are C 1-4 Alkyl, C 1-4 haloalkyl, 5-7 membered heteroaryl or C 6-7 Aryl.

3. The compound according to claim 2, wherein Ring A is 4. The compound according to claim 3, wherein Ring A is selected from 5. The compound according to any one of claims 1 to 4, wherein The rings D, D1, and D2 are each independently selected from the following compounds:

6. The compound according to any one of claims 1 to 5, wherein The compounds of formula (I) and formula (ID) are selected from:

7. Use of the compound of formula (I) or (II) as claimed in any one of claims 1 to 6 in hemicyanine dyes.

8. Use of the compound of formula (I) or (II) as claimed in any one of claims 1 to 6 in fluorescence and photoacoustic imaging.

9. Use of the compound of formula (I) or (II) as claimed in claim 8 in integrated photodiagnosis and therapy.

10. Use of the compound of formula (I) or (II) as claimed in claim 9 in an integrated phototherapy preparation.

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