Near-infrared second-region cyanine dye molecule, preparation method and application

Abstract

The invention relates to the technical field of organic functional materials, in particular to a near-infrared second-region cyanine dye molecule as well as a preparation method and application thereof, the structural formula of the near-infrared second-region cyanine dye molecule is shown as a formula I or a formula II: # imgabs0 #, the cyanine dye molecule is modified by benzopyranium, the excitation wavelength and the fluorescence emission wavelength of the near-infrared second-region cyanine dye molecule both reach the near-infrared second region, and the near-infrared second-region cyanine dye molecule is a near-infrared second-region cyanine dye molecule. Strong absorption and emission are shown in a near-infrared second region, and the Stokes shift is greater than 50nm; the synthetic method of the fluorescent dye is simple, raw materials are easy to obtain, and the yield is high; the cyanine dye disclosed by the invention is excellent in photo-thermal stability and good in photo-thermal imaging effect, has a good phototherapy effect and has a great potential application prospect in the field of tumor treatment.

Description

Technical Field

[0001] The invention relates to the technical field of organic functional materials, and in particular to a near-infrared second-zone cyanine dye molecule, a preparation method and an application thereof. Background Art

[0002] Breast cancer is one of the malignant tumors that seriously threatens women's life and health. According to the latest statistics, the incidence and mortality of breast cancer worldwide are 29% and 15% respectively. In China, the incidence of breast cancer has shown a significant upward trend in recent years and has ranked first among female cancers. Clinical treatments for breast cancer include surgery, radiotherapy, chemotherapy and immunotherapy, but these methods have their own limitations. For example, chemotherapy can easily cause breast cancer cells to develop multidrug resistance, reduce drug efficacy, and most chemotherapy drugs lack selectivity and have toxic side effects that cannot be ignored. Therefore, the current clinical treatment of breast cancer still cannot avoid late metastasis and recurrence in about 30% of patients. It can be seen that exploring safe and effective treatment methods and strategies has extremely important theoretical significance and application value for the treatment of clinical breast cancer.

[0003] Phototherapy is an advanced, non-invasive treatment for tumors, which mainly includes two categories, photothermal therapy (PTT) and photodynamic therapy (PDT). Photothermal therapy is an emerging treatment method that uses photothermal converters (PTAs) to absorb light energy under the irradiation of a remote excitation light source and convert light into heat energy, thereby causing tumor cells to reach a certain temperature and cause their death. This treatment method has the advantages of being controllable in time and space, and having low damage to surrounding normal tissues. In recent years, it has attracted widespread attention in the field of tumor research. Among the widely reported PTAs, such as precious metal nanoparticles, carbon nanomaterials, semiconductor polymers and conjugated small molecules, heptamethine cyanine dye (Cy7) has strong near-infrared (NIR) absorption, good photothermal conversion efficiency (PCE) and excellent biocompatibility, and is considered to be an ideal organic PTA. The easy structural modification of Cy7 dye helps to develop multifunctional PTAs with superior photothermal performance, showing great potential for clinical transformation. However, its weak water solubility and poor optical stability in hydrophilic environments limit its application in the field of biology. Therefore, developing a near-infrared photosensitizer with near-infrared emission and high photothermal stability is of great significance to improving the effect of photothermal therapy. Summary of the invention

[0004] In order to overcome the shortcomings of the above technical defects, the present invention provides a near-infrared II cyanine dye molecule, a preparation method and an application, so as to achieve the purpose of absorption and emission in the NIR-Ⅱ region, high stability and excellent photothermal performance.

[0005] In order to achieve the above object, the technical solution of the present invention is:

[0006] In one aspect, a near-infrared second-zone cyanine dye molecule is provided, the structural formula of which is shown in Formula I or Formula II:

[0007]

[0008] Furthermore, the maximum emission wavelength of the cyanine dye molecule is 1050-1150 nm, and the maximum absorption wavelength is 1000-1100 nm.

[0009] In a second aspect, a method for preparing a near-infrared second-zone cyanine dye molecule is provided, wherein a compound of formula III undergoes a condensation reaction with a compound of formula IV to obtain a cyanine dye FN-7, the structural formula of which is shown in formula I;

[0010] Or the compound of formula III reacts with the compound of formula V to produce cyanine dye FM-7, the structural formula of which is shown in formula II;

[0011] The structures of the compounds are as follows:

[0012]

[0013] Furthermore, the condensation reaction conditions are: reacting at 80-100° C. for 5-7 hours, cooling to room temperature, removing ethanol, and subjecting the obtained solid to column chromatography.

[0014] Furthermore, the condensation reaction is carried out in the presence of an organic solvent and an activator;

[0015] The organic solvent is selected from one or more of anhydrous ethanol, anhydrous methanol, N,N-dimethylformamide or acetic anhydride;

[0016] The activator is selected from one or more of potassium carbonate, sodium acetate and potassium acetate.

[0017] Furthermore, the synthetic route of the compound of formula IV is as follows:

[0018]

[0019] Furthermore, the synthesis route of the compound of formula V is as follows:

[0020]

[0021] In a third aspect, a method is provided for preparing tumor photothermal therapy and / or diagnostic reagents or drugs based on the above-mentioned cyanine dye molecules and the cyanine dye molecules prepared by the above-mentioned method.

[0022] In a fourth aspect, a method for applying the cyanine dye molecule prepared by the above method in medical imaging is provided, wherein the medical imaging includes bioluminescence imaging and / or photoacoustic imaging.

[0023] In a fifth aspect, a pharmaceutical composition is provided, comprising the above-mentioned cyanine dye molecule, the cyanine dye molecule obtained by the above-mentioned method, or a pharmaceutically acceptable salt, solvate, enantiomer, diastereomer, or tautomer thereof.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) The synthesis method of the benzopyranium-modified near-infrared second-zone cyanine dyes (FN-7 and FM-7) of the present invention has simple steps and high yield;

[0026] (2) A benzopyranium-modified near-infrared zone II cyanine dye (FN-7 and FM-7) of the present invention exhibits excellent photostability with negligible attenuation after continuous laser irradiation at 808 nm for 10 min, and high photothermal stability is observed even after four heating-cooling cycles;

[0027] (3) The present invention provides a new near-infrared second-zone cyanine dye (FN-7 and FM-7) with a high photothermal temperature, achieving very good photothermal effects on mouse tumors;

[0028] (4) The benzopyranium-modified near-infrared second-zone cyanine dyes (FN-7 and FM-7) of the present invention have good photothermal imaging properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The invention discloses a near-infrared second-zone cyanine dye FN-7 and FM-7 modified by benzopyranium 1 HNMR spectrum;

[0030] Figure 2 HRMS spectra of the benzopyrylium-modified near-infrared second-zone cyanine dyes FN-7 and FM-7 of the present invention;

[0031] Figure 3 The ultraviolet fluorescence spectra of the near-infrared second-zone cyanine dyes FN-7 and FM-7 modified by benzopyrylium of the present invention;

[0032] Figure 4 The temperature changes of different concentrations of the near-infrared second-zone cyanine dyes FN-7 and FM-7 modified by benzopyranium of the present invention;

[0033] Figure 5 The temperature changes of different powers of the near-infrared second-zone cyanine dyes FN-7 and FM-7 modified by benzopyrylium of the present invention;

[0034] Figure 6 The stability changes of the near-infrared second-zone cyanine dyes FN-7 and FM-7 modified by benzopyrylium of the present invention;

[0035] Figure 7 The photothermal conversion efficiency of the near-infrared second-zone cyanine dyes FN-7 and FM-7 modified by benzopyrylium of the present invention;

[0036] Figure 8 Live and dead cell staining of the near-infrared second-zone cyanine dye FN-7 modified by benzopyranium of the present invention;

[0037] Fig. 9 The invention discloses photothermal imaging of the second-zone near-infrared cyanine dyes FN-7 and FM-7 modified by benzopyranium in mice. DETAILED DESCRIPTION

[0038] In order to make those skilled in the art better understand the technical scheme of the present invention, the present invention is described in detail below in conjunction with specific embodiments. The experimental methods for which specific conditions are not indicated in the following examples are usually carried out under normal conditions or according to the conditions recommended by the manufacturer. The test materials used in the following examples, unless otherwise specified, are purchased from conventional biochemical reagent stores. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, all professional and scientific terms used in the text have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar to or equal to the recorded content can all be applied to the present invention. The preferred implementation methods and materials described in the text are only for demonstration purposes.

[0039] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0040] Based on the present invention, the present invention aims to provide a benzopyrylium-modified cyanine dye, which has high photothermal stability in the near-infrared second region (NIR-II region), improves the photothermal ablation ability of tumors, causes thermal damage to the nuclear genetic material of tumor cells, and induces acute necrosis, cell apoptosis and microenvironment destruction of tumors.

[0041] In some specific embodiments, a near-infrared second-zone cyanine dye molecule is provided, and its structural formula is shown in Formula I:

[0042]

[0043] In another specific embodiment, a preparation method based on the above-mentioned cyanine dye molecule is provided, comprising the following steps:

[0044] S1. Dissolve 1-[4-(diethylamino)-2-hydroxy]acetophenone and dimethylaminoacetophenone in acetic acid, heat in perchloric acid, and cool to obtain compound 1, the structural formula of which is shown in Formula IV;

[0045]

[0046] S2. The obtained compound 1 is subjected to a condensation reaction with N-[(3-(anilinomethylene)-2-chloro-1-cyclohexene-1-yl)methylene]aniline hydrochloride, cooled after the reaction, and subjected to column chromatography to obtain a cyanine dye FN-7, the structural formula of which is shown in Formula I;

[0047]

[0048] It is understood that the condensation reaction is carried out in the presence of an organic solvent and an activating agent;

[0049] The organic solvent is selected from one or more of anhydrous ethanol, anhydrous methanol, N,N-dimethylformamide or acetic anhydride;

[0050] The activator is selected from one or more of potassium carbonate, sodium acetate and potassium acetate.

[0051] In some examples of this embodiment, in S1, the specific steps are: dissolving 1-[4-(diethylamino)-2-hydroxy]acetophenone and dimethylaminoacetophenone in acetic acid, heating to reflux in perchloric acid at 120° C. for 12 h, cooling to room temperature, and adding distilled water to the reaction bottle to obtain compound 1.

[0052] In some examples of this embodiment, the molar ratio of 1-[4-(diethylamino)-2-hydroxy]acetophenone to dimethylaminoacetophenone is 1:1.2.

[0053] In some examples of this implementation mode, in S2, the specific steps are: dissolving compound 1 and N-[(3-(anilinomethylene)-2-chloro-1-cyclohexen-1-yl)methylene]aniline hydrochloride in ethanol, heating at 90° C. for 6 hours, and then cooling to room temperature, removing ethanol and drying by column chromatography to obtain cyanine dye FN-7.

[0054] In some examples of this embodiment, the molar ratio of the compound 1 to N-[(3-(anilinomethylene)-2-chloro-1-cyclohexen-1-yl)methylene]aniline hydrochloride is 2.1:1, and the eluent is a mixture of dichloromethane and methanol in a volume ratio of 80:1.

[0055] In some specific embodiments, a near-infrared second-zone cyanine dye molecule is provided, and its structural formula is shown in Formula II:

[0056]

[0057] In another specific embodiment, a preparation method based on the above-mentioned cyanine dye molecule is provided, comprising the following steps:

[0058] S1: dissolving 1-[4-(diethylamino)-2-hydroxy]acetophenone and 4-tetraphenylethylene benzoyl in acetic acid and heating in perchloric acid to obtain compound 2, the structural formula of which is shown in Formula IV;

[0059]

[0060] S2. The obtained compound 2 is subjected to a condensation reaction with N-[(3-(anilinomethylene)-2-chloro-1-cyclohexene-1-yl)methylene]aniline hydrochloride, cooled after the reaction, and subjected to column chromatography to obtain a photosensitizer FM-7, the structural formula of which is shown in Formula II.

[0061]

[0062] It is understood that the condensation reaction is carried out in the presence of an organic solvent and an activating agent;

[0063] The organic solvent is selected from one or more of anhydrous ethanol, anhydrous methanol, N,N-dimethylformamide or acetic anhydride;

[0064] The activator is selected from one or more of potassium carbonate, sodium acetate and potassium acetate.

[0065] In some examples of this implementation mode, in S1, the specific steps are: dissolving 1-[4-(diethylamino)-2-hydroxy]acetophenone and 4-tetraphenylethylene benzoyl in acetic acid, heating to reflux in perchloric acid at 120°C for 12 hours, cooling to room temperature, and adding distilled water to the reaction bottle to obtain compound 2.

[0066] In some examples of this embodiment, the molar ratio of 1-[4-(diethylamino)-2-hydroxy]acetophenone to 4-tetraphenylethylene benzoyl is 1:1.2.

[0067] In some examples of this implementation mode, in S2, the specific steps are: dissolving the obtained compound 2 and N-[(3-(anilinomethylene)-2-chloro-1-cyclohexen-1-yl)methylene]aniline hydrochloride in ethanol, heating at 90°C for 6 hours and then cooling to room temperature, removing excess ethanol and drying by column chromatography to obtain cyanine dye FM-7.

[0068] In some examples of this embodiment, in S2, the molar ratio of the compound 2 to N-[(3-(anilinomethylene)-2-chloro-1-cyclohexen-1-yl)methylene]aniline hydrochloride is 2.1:1, and the eluent is a mixture of dichloromethane and methanol in a volume ratio of 50:1.

[0069] The technical solution of the present invention is further described in detail below in conjunction with specific embodiments and drawings. It should be understood that the following embodiments are only used to explain the present invention and are not used to limit the present invention.

[0070] Example 1 Synthesis of cyanine dye FN-7

[0071] (1) Dissolve 1-(4-(diethylamino)-2-hydroxyacetophenone (0.2 g, 1 mmol) and 4-dimethylbenzoyl (0.163 g, 1.2 mmol) in 20 mL of acetic acid and 25 mL of HClO4. Heat under reflux at 125°C for 15 h, cool to room temperature, add to ice water to obtain a bright green solid, and dry. Dissolve in dichloromethane and spin dry to obtain a black solid. Proceed directly to the next step. 1 H NMR (500MHz, CDCl3) δ8.14(t,J=6.2Hz,2H),7.81(d,J=9.4Hz,1H),7.52(s,1H),7.07(dd,J=9.4,2.5Hz,1H),6.9 3(d,J=2.5Hz,1H),6.80(d,J=9.3Hz,2H),3.62(q,J=7.2Hz,4H),3.18(s,6H),2.75(s,3H),1.35(t,J=7.2Hz,6H).

[0072] (2) Compound 1 (0.47 g, 0.8 mmol), N-[(3-(anilinomethylene)-2-chloro-1-cyclohexen-1-yl)methylene]aniline hydrochloride (0.185 g, 0.4 mmol) and sodium acetate (0.076 g) were dissolved in anhydrous ethanol and reacted at 90° C. for 6 h. After cooling to room temperature, the ethanol was removed and the obtained solid was subjected to column chromatography (eluent: dichloromethane:methanol=50:1) to obtain a black solid, which was characterized by mass spectrometry and nuclear magnetic resonance ( Figure 1-2 ). 1HNMR(500MHz,DMSO)δ8.10(d,J=13.6Hz,2H),7.97(dd,J=24.6,9.2Hz,6H),7.35(s,2H),6.89–6.84(m,4H),6.80(d,J=9.1Hz,4 H),6.74(d,J=2.1Hz,2H),3.52(d,J=7.0Hz,8H),3.09(d,J=19.1Hz,12H),2.75(s,4H),1.87(s,2H),1.20(t,J=7.0Hz,12H).[M + ]:calcd for[C 52 H 58 CIN4O2] + :805.4243; found805.4168.

[0073] The synthetic route is as follows:

[0074]

[0075] Example 2 Synthesis of Cyanine Dye FM-7

[0076] (1) Dissolve 1-(4-(Diethylamino)-2-hydroxyacetophenone (0.2 g, 1 mmol) and 4-tetraphenylethylene benzoyl (0.4 g, 1.2 mmol) in 20 mL of acetic acid and 25 mL of HClO4. Heat under reflux at 125°C for 15 h, cool to room temperature, and add water to obtain a pink solid. Dry the solid and proceed directly to the next step.

[0077] (2) Compound 2 (0.5 g, 0.8 mmol), N-[(3-(anilinomethylene)-2-chloro-1-cyclohexen-1-yl)methylene]aniline hydrochloride (0.132 g, 0.4 mmol) and sodium acetate (0.076 g) were dissolved in anhydrous ethanol and reacted at 90° C. for 6 h. After cooling to room temperature, the ethanol was removed and the obtained solid was subjected to column chromatography (eluent: dichloromethane:methanol=50:1) to obtain a black solid, which was characterized by mass spectrometry and nuclear magnetic resonance ( Figure 1-2 ). 1H NMR (500MHz, CDCl3) δ8.13(d,J=13.7Hz,2H),7.95(d,J=9.4Hz,2H),7.74(d,J=8.1Hz,4H),7.23(d,J=8.1Hz,4H),7.21–7.11(m,24H) ,7.08(d,J=2.9Hz,8H),6.93(t,J=12.0Hz,4H),6.52(s,2H),3.53–3.48(m,8H),2.82(s,4H),2.01(s,2H),1.29(t,J=7.0Hz,12H).[M + ]:calcd for[C 88 H 76 CIN2O2] + :1229.0375; found1228.5585.

[0078] Synthesis route:

[0079]

[0080] Example 3 Investigation of the UV Fluorescence Spectra of Benzopyranium-Modified Near-Infrared Zone II Cyanine Dyes

[0081] The UV and fluorescence properties of the molecule synthesized in Example 1 were tested in dichloromethane. The spectrum is shown in FIG. Figure 3 As shown. The maximum UV absorption peaks of FN-7 and FM-7 are 1044nm and 1050nm, respectively, and the maximum fluorescence emission peaks are 1094nm and 1102nm, respectively. The results show that both FN-7 and FM-7 exhibit strong absorption and emission in the near-infrared region II. The Stokes shifts of FN-7 and FM-7 reach 50nm and 52nm, respectively.

[0082] Example 4 Investigation of the Photothermal Properties of Benzopyranium-Modified Near-Infrared Zone II Cyanine Dyes

[0083] At a fixed laser power (808 nm, 1.5 W / cm 2 ), as the concentration of cyanine dye increases, the temperature of FN-7 and FM-7 solutions also increases. Figure 4 As shown in the figure, when the concentration of cyanine dye increases from 20μM to 80μM, the temperature of FN-7 increases from 50.5 to 77.8℃ within 600s, with a temperature increase of 27.3℃, and the temperature of FM-7 increases from 47.1 to 74.3℃, with a temperature increase of 27.2℃. The temperature difference shows that the temperature of FN-7 is the highest, which is 77.8℃. The results show that cyanine dyes have excellent photothermal properties and FN-7 has the best photothermal properties.

[0084] like Figure 5 As shown, as the power density increases from 0.8W / cm 2 Increase to 1.5W / cm 2 , the temperature of cyanine dye FN-7 increased from 50.7℃ to 77.8℃, and the temperature of FM-7 increased from 46.1℃ to 74.3℃.

[0085] like Figure 6 As shown, after four photothermal cycles of 808nm laser irradiation, the temperature of cyanine dye FN-7 stabilized at 72.4°C and the temperature of FM-7 stabilized at 71°C. The results show that the two cyanine dyes exhibit excellent photothermal stability after 808nm continuous laser irradiation for 10 minutes.

[0086] At room temperature, using 808nm (1.5W / cm 2 ) irradiated PBS solutions (80 μM) of cyanine dyes FN-7 and FM-7. Figure 7 As shown, the photothermal conversion efficiency of FN-7 is 68%, while that of FM-7 is 66.2%. The results show that FN-7 and FM-7 have good photothermal performance and have great potential application prospects.

[0087] Example 5 Investigation of the Cytotoxicity of Benzopyranium-Modified Near-Infrared Zone II Cyanine Dye FN-7

[0088] To more intuitively display the phototherapy performance of FN-7, we used calcein AM (green) and propidium iodide (red) dyes for live and dead cell staining. Green fluorescence indicates live cells, and red indicates dead cells. An 808 nm laser (2 W / cm 2 ) irradiated PBS, PBS+laser, FN-7, and FN-7+laser solutions for 15 min. In the dark, a small amount of cell death was observed in the FN-7 group, indicating that the cytotoxicity of FN-7 was low and it had good biocompatibility. After laser irradiation, HeLa cells treated with FN-7 showed obvious red fluorescence, indicating that most of the HeLa cells died, such as Figure 8 The results show that at a power intensity (2W / cm 2 ) laser irradiation, FN-7 can effectively kill cancer cells and has a good phototherapy effect, which has great potential application prospects in the field of tumor treatment.

[0089] Example 6 In vivo photothermal imaging of benzopyranium-modified near-infrared zone II cyanine dyes FN-7 and FM-7

[0090] Two photosensitizers (100 μM, 200 μL, PBS solvent) were injected subcutaneously at the thigh root of BABL / C nude mice or mice, and the light was irradiated at 808 nm, 1 W / cm 2The laser was irradiated for 10 min and dynamic monitoring was performed. Fig. 9 As shown in the figure, the local temperature of mice injected with FN-7 solution increased significantly under laser irradiation, with the highest temperature reaching 52°C. The local temperature of mice injected with FM-7 solution increased to 48°C under laser irradiation. The results show that the near-infrared second-zone cyanine dyes FN-7 and FM-7 modified by benzopyranium have good photothermal imaging effects. This has potential application prospects in the field of photothermal therapy of tumors.

[0091] Finally, it should be noted that the above description is only a preferred embodiment of the present invention. Under the guidance of the present invention, those skilled in the art can make a variety of similar expressions without violating the purpose and claims of the present invention, and such changes all fall within the protection scope of the present invention.

Claims

1. A near-infrared second-zone cyanine dye molecule, characterized in that: Its structural formula is shown in Formula I or Formula II:

2. The near-infrared second-zone cyanine dye molecule according to claim 1, characterized in that: The maximum emission wavelength of the cyanine dye molecule is 1050-1150 nm, and the maximum absorption wavelength is 1000-1100 nm.

3. The method for preparing the near-infrared second-zone cyanine dye molecule according to claim 1 or 2, characterized in that: The compound of formula III undergoes a condensation reaction with the compound of formula IV to obtain a cyanine dye FN-7, the structural formula of which is shown in formula I; Or the compound of formula III reacts with the compound of formula V to produce cyanine dye FM-7, the structural formula of which is shown in formula II; The structures of the compounds are as follows:

4. The method according to claim 3, characterized in that The condensation reaction conditions are: reacting at 80-100° C. for 5-7 hours, cooling to room temperature, removing ethanol, and performing column chromatography on the obtained solid to obtain the product.

5. The method according to claim 4, characterized in that The condensation reaction is carried out in the presence of an organic solvent and an activator; The organic solvent is selected from one or more of anhydrous ethanol, anhydrous methanol, N,N-dimethylformamide or acetic anhydride; The activator is selected from one or more of potassium carbonate, sodium acetate and potassium acetate.

6. The method according to claim 3, characterized in that: The synthetic route of the compound of formula IV is as follows:

7. The method according to claim 3, characterized in that The synthetic route of the compound of formula V is as follows:

8. Use of the cyanine dye molecule according to claim 1 or 2, or the cyanine dye molecule prepared by the method according to any one of claims 3 to 7 in the preparation of tumor photothermal therapy and / or diagnostic reagents or drugs.

9. Use of the cyanine dye molecule according to claim 1 or 2 or the cyanine dye molecule prepared by the method according to any one of claims 3 to 7 in medical imaging, wherein the medical imaging includes bioluminescence imaging and / or photoacoustic imaging.

10. A pharmaceutical composition, characterized in that Contains the cyanine dye molecule according to claim 1 or 2, the cyanine dye molecule prepared by the method according to any one of claims 3 to 7, or its pharmaceutically acceptable salt, solvate, enantiomer, diastereomer, tautomer.

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