Preparation method and application of near-infrared two-window fluorescence contrast agent

Abstract

The invention relates to the field of biochemical materials and organic photoelectric information materials, and discloses a near-infrared two-window fluorescent contrast agent which has remarkable advantages in the field of near-infrared two-window fluorescent imaging, can monitor dynamic change of molecular level in a living body in real time, and has a good application prospect in the field of near-infrared two-window fluorescent imaging. The characteristic is helpful for early detection and timely treatment of diseases. In the aspect of drug research, the contrast agent can be used for tracking distribution, metabolic pathways and action targets of drugs in organisms, so that the curative effect and potential toxic and side effects of the drugs are accurately evaluated. In a clinical treatment link, the system can monitor the disease progress and the treatment effect in real time, provides a key basis for scientific formulation of a personalized treatment scheme, and assists in improving the accuracy and effectiveness of medical services.

Description

Technical Field

[0001] The invention relates to the fields of biochemical materials and organic optoelectronic information materials, and in particular to a preparation method and application of a near-infrared two-window fluorescent contrast agent. Background Art

[0002] Traditional visible light and near-infrared region I (NIR-I, 700-900nm) fluorescence imaging technologies have certain limitations, such as limited tissue penetration depth, insufficient spatial resolution, and large interference from biological tissue autofluorescence. As the requirements for in vivo imaging continue to increase, it is necessary to develop technologies that can perform imaging at deeper tissue depths and higher resolutions. Near-infrared region II fluorescence imaging technology has emerged, and near-infrared second-window fluorescent contrast agents are the core elements of this technology.

[0003] Near-infrared two-zone fluorescence imaging technology has unique advantages in biomedical imaging. Near-infrared two-window fluorescence imaging reduces light scattering and absorption, allowing light to penetrate deeper tissues and achieve imaging of deep tissue organs such as the brain and liver. It also has higher temporal and spatial resolution, reduced scattering and reduced autofluorescence, allowing near-infrared two-zone fluorescence imaging to provide higher spatial resolution, clearly display fine structures such as tiny blood vessels and tumor boundaries, and achieve real-time monitoring of changes at the molecular level in the body, which helps to buy more time for early detection and treatment of diseases. Summary of the invention

[0004] In order to solve the problems in the prior art, the present invention prepares a near-infrared two-window fluorescent contrast agent. It can be used for near-infrared two-window fluorescent imaging to achieve real-time monitoring of changes in molecular levels in the body, so as to detect diseases earlier and start treatment. It can also be used to track the distribution, metabolism and target of drugs in the body, and evaluate drug efficacy and toxic side effects. During the treatment process, the development of the disease and the treatment effect can be monitored in real time, providing a basis for the formulation of personalized treatment plans.

[0005] The purpose of the present invention is achieved through the following technical solutions:

[0006] The present invention provides a near-infrared two-window fluorescent contrast agent, the structural formula of which is:

[0007]

[0008] Wherein, X is any one of F, Cl, Br, and I;

[0009] W is any one of O, S and Se;

[0010] R is hydrogen or methyl, ethyl, trifluoromethyl, an alkyl group having 3 to 8 carbon atoms, an aryl derivative, or an alkoxy chain having 3 to 8 carbon atoms and oxygen atoms.

[0011] In some embodiments, R is hydrogen or any one of methyl, ethyl, trifluoromethyl, an alkyl group having 3 to 8 carbon atoms, an aryl derivative, and an alkoxy chain having 3 to 8 carbon atoms and oxygen atoms.

[0012] In a preferred embodiment, R is methyl.

[0013] Furthermore, the near-infrared two-window fluorescent contrast agent is selected from the following structures:

[0014]

[0015] In a second aspect, the present invention provides a method for preparing the near-infrared two-window fluorescent contrast agent described in the first aspect, wherein the synthesis of formula 6 comprises the following steps:

[0016] S1, compound 1 reacts with ethyl bromide / ethyl iodide and potassium carbonate / sodium in N,N-dimethylformamide / acetonitrile / dimethyl sulfoxide solution to obtain compound 2;

[0017] S2, compound 2, 5-bromothiophene-2-carboxaldehyde and potassium / sodium hydroxide in an ethanol solution to react to obtain compound 3;

[0018] S3 and compound 3 react in a mixed solution of nitromethane and methanol / ethanol to obtain compound 4;

[0019] S4, compound 4 reacts in a mixed solution of ammonium acetate and n-butanol / ethanol / acetic anhydride to obtain compound 5;

[0020] S5, compound 5 in a mixed solution of dichloromethane, boron trifluoride etherate and diisopropylethylamine / triethylamine to obtain compound 6;

[0021] Among them, compound 1 is Compound 2 is

[0022] Compound 3 is Compound 4 is

[0023] Compound 5 is Compound 6 is

[0024] R, W, X are as defined in any one of claims 1 to 3

[0025] Further, in step S1, in N,N-dimethylformamide / acetonitrile / dimethyl sulfoxide solution, compound 1 reacts with ethyl bromide / ethyl iodide and potassium carbonate / sodium in equimolar amounts at room temperature overnight to obtain compound 2;

[0026] In step S2, compound 2 reacts with 5-bromothiophene-2-carboxaldehyde and potassium hydroxide / sodium hydroxide in an ethanol solution to obtain compound 3;

[0027] In step S3, in a mixed solution of nitromethane and methanol / ethanol, compound 3 is reacted under reflux (70° C.) (24 h) to obtain compound 4;

[0028] In step S4, compound 4 is refluxed in a mixed solution of ammonium acetate and n-butanol / ethanol / acetic anhydride at room temperature overnight to obtain compound 5;

[0029] In step S5, compound 5 is reacted in a mixed solution of dichloromethane, boron trifluoride etherate and diisopropylethylamine / triethylamine at room temperature (4 h) to obtain compound 6;

[0030] In a third aspect, the present invention provides the use of the near-infrared dual-window fluorescent contrast agent described in the first aspect in disease diagnosis and treatment monitoring.

[0031] Specifically, under near-infrared light excitation conditions, contrast agents can perform real-time, non-invasive, high-resolution observations of physiological and pathological processes in living animals and the human body, such as early detection and localization of tumors, monitoring of angiogenesis, and distribution and metabolism of drugs in the body.

[0032] The beneficial effects of the present invention are:

[0033] The present invention prepares a near-infrared two-window fluorescent contrast agent. The contrast agent has significant advantages in the field of near-infrared two-window fluorescent imaging, and can implement real-time monitoring of dynamic changes at the molecular level in the body. This feature contributes to the early detection and timely treatment of diseases. In terms of drug research, the contrast agent can be used to track the distribution, metabolic pathways and targets of drugs in the body, and then accurately evaluate the efficacy and potential toxic side effects of drugs. In the clinical treatment stage, it can monitor the progression of the disease and the treatment effect in real time, provide a key basis for the scientific formulation of personalized treatment plans, and help improve the accuracy and effectiveness of medical services.

[0034] The design and synthesis of the contrast agent of the present invention are supported by theoretical basis, and the preparation method is simple, the synthesis conditions are mild, and the raw materials are abundant, which has important guiding significance for the design of multifunctional near-infrared biological contrast agents in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0036] Figure 1 is the mass spectrum of compound Ⅰ in the embodiment of the present invention;

[0037] Figure 2 is the compound I in the embodiment of the present invention 1 H-NMR spectrum;

[0038] Figure 3 is the compound I in the embodiment of the present invention 19 F-NMR spectrum;

[0039] Figure 4 is the compound I in the embodiment of the present invention 13 C-NMR spectrum;

[0040] Figure 5 is the absorption spectrum of compound I in the embodiment of the present invention;

[0041] Figure 6 is the fluorescence spectrum of compound I in the embodiment of the present invention;

[0042] Figure 7 This is a near-infrared two-window fluorescence imaging image of compound I in the example of the present invention in mice. DETAILED DESCRIPTION

[0043] It should be noted that the following detailed descriptions are illustrative and are intended to provide further explanation of the present application. Unless otherwise specified, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art to which the present application belongs.

[0044] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the art can be used. The raw materials or instruments used are all conventional products that can be obtained commercially, including but not limited to the raw materials or instruments used in the examples of this application.

[0045] The yield calculation method of the present invention is "product (mol) / reaction substrate (mol)*100%". In the present invention, a mass spectrograph is used to detect the composition of the substance. A nuclear magnetic resonance spectrometer (NMR) is used to characterize the chemical structure of the target compound. An ultraviolet spectrophotometer (UV-3600UV-vis-NIRspectrophotometer) and a steady-state / transient fluorescence spectrometer (FLS-920spectrometer) are used to test the absorption and emission spectra of the compounds, respectively. A near-infrared thermal imager (E40) is used to test the temperature changes of compounds A-1 and A-2.

[0046] Embodiment 1:

[0047] The chemical structure and synthesis route of the near-infrared two-window fluorescent contrast agent compound I are as follows:

[0048]

[0049] Synthesis of compound 2':

[0050] A mixture of compound 1' (20 mmol), bromoethane (20 mmol) and potassium carbonate (20 mmol) was added to 10 mL of N,N-dimethylformamide solution. The mixture was then stirred at room temperature overnight. The final compound 2' was obtained by silica gel chromatography (PE:EA=15:1) (yield: 53%).

[0051] Synthesis of compound 3':

[0052] A mixture of compound 2' (10 mmol), 5-bromothiophene-2-carboxaldehyde (10 mmol) and potassium hydroxide (10 mmol) was stirred at 70°C for 24 h in an ethanol solution, and the crude product was obtained by vacuum distillation to remove the solvent. Compound 3' (yield: 76%) was obtained by column chromatography (PE:EA=10:1).

[0053] Synthesis of compound 4':

[0054] A mixture of compound 3' (10 mmol), methanol (5 mL) and nitromethane (10 mL) was stirred at 70°C for 24 h, and the crude product was obtained by vacuum distillation to remove the solvent. Compound 4' was obtained by column chromatography (PE:EA=10:1) (yield: 88%).

[0055] Synthesis of compound 5':

[0056] Compound 4' was added to 10 mL of a mixed solution of ammonium acetate and n-butanol, and the mixture was stirred at room temperature overnight. The final compound 5' was obtained by silica gel chromatography (PE:EA=15:1) (yield: 18%).

[0057] Synthesis of Compound I:

[0058] Compound 5' was added to a mixed solution of dichloromethane, boron trifluoride etherate and diisopropylamine (10 mL), and the mixture was stirred at room temperature for 4 h. The final compound I was obtained by silica gel chromatography (PE:EA=15:1) (yield: 35%).

[0059] Compound I: 1H NMR (400MHz, CDCl3): δ (ppm) = 8.06 (d, J = 8.8Hz, 4H), 7.52 (d, J = 4.0Hz, 2H), 7.10 (d, J = 4. 0Hz, 2H), 6.92 (s, 2H), 6.71 (d, J = 8.8Hz, 4H), 3.46-3.41 (m, 8H), 1.22 (t, J = 7.2Hz, 12H).

[0060] 19 F NMR (377MHz, CDCl3): δ (ppm) = -131.83 (q, 2F).

[0061] 13 C NMR (100MHz, CDCl3): δ (ppm) = 156.04, 149.64, 136.70, 133.66, 131.92, 130.51, 127.94, 118.18, 116.03, 115.27, 111.36, 44.59, 12.79.

[0062] The performance of the compound obtained in Example 1 was verified, and the results were as follows Figure 1-6 shown.

[0063] By mass spectrometry ( Figure 1 ) and nuclear magnetic resonance hydrogen, carbon, and fluorine spectra ( Figures 2 to 4 ) proved the correctness of compound I.

[0064] By absorption spectroscopy ( Figure 5 ), proving that compound I has near-infrared second window absorption.

[0065] By fluorescence spectroscopy ( Figure 6 ), proving that compound I has near-infrared second window emission.

[0066] Through mouse infrared two-window fluorescence imaging ( Figure 7 ), demonstrating that compound I has the properties of a near-infrared two-window fluorescence imaging contrast agent in mice.

[0067] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A near-infrared two-window fluorescent contrast agent, characterized in that: Its structural formula is: Wherein, X is any one of F, Cl, Br, and I; W is any one of O, S and Se; R is hydrogen or methyl, ethyl, trifluoromethyl, an alkyl group having 3 to 8 carbon atoms, an aryl derivative, or an alkoxy chain having 3 to 8 carbon atoms and oxygen atoms.

2. The near-infrared double-window fluorescent contrast agent according to claim 1, characterized in that: R is hydrogen or methyl, ethyl, trifluoromethyl, an alkyl group having 3 to 8 carbon atoms, an aryl derivative, or an alkoxy chain having 3 to 8 carbon atoms and oxygen atoms.

3. The near-infrared double-window fluorescent contrast agent according to claim 1, characterized in that: R is methyl.

4. The near-infrared double-window fluorescent contrast agent according to claim 1, characterized in that: Select from the following structures:

5. The method for preparing the near-infrared double-window fluorescent contrast agent according to any one of claims 1 to 4, characterized in that: The following steps are involved: S1, compound 1 reacts with ethyl bromide / ethyl iodide and potassium carbonate / sodium in N,N-dimethylformamide / acetonitrile / dimethyl sulfoxide solution to obtain compound 2; S2, compound 2, 5-bromothiophene-2-carboxaldehyde and potassium hydroxide / sodium hydroxide in an ethanol solution to react to obtain compound 3; S3 and compound 3 react in a mixed solution of nitromethane and methanol / ethanol to obtain compound 4; S4, compound 4 reacts in a mixed solution of ammonium acetate and n-butanol / ethanol / acetic anhydride to obtain compound 5; S5, compound 5 in a mixed solution of dichloromethane, boron trifluoride etherate and diisopropylethylamine / triethylamine to obtain compound 6; Among them, compound 1 is Compound 2 is Compound 3 is Compound 4 is Compound 5 is Compound 6 is 6. Use of the near-infrared dual-window fluorescent contrast agent according to any one of claims 1 to 4 in disease diagnosis and treatment monitoring.

7. Use of the near-infrared two-window fluorescent contrast agent according to claim 6 in disease diagnosis and treatment monitoring, characterized in that; Under near-infrared light excitation conditions, contrast agents can provide real-time, non-invasive, high-resolution observation of physiological and pathological processes in living animals and humans, such as early detection and localization of tumors, monitoring of angiogenesis, and distribution and metabolism of drugs in the body.

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