Nitroreductase-responsive diagnostic and therapeutic integrated probe and its preparation method and application

A technology of reductase and nitro, which is applied in the field of biomedicine, can solve the problems of lack of effective treatment methods and difficulties in tumor diagnosis, and achieve the effects of easy promotion and application, good biological safety, and strong tissue penetration

Active Publication Date: 2022-04-05
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the above problems, the present invention provides a nitroreductase-responsive integrated probe for diagnosis and treatment and its preparation method, aiming at solving the technical problems of difficulty in early tumor diagnosis and lack of effective treatment means

Method used

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  • Nitroreductase-responsive diagnostic and therapeutic integrated probe and its preparation method and application
  • Nitroreductase-responsive diagnostic and therapeutic integrated probe and its preparation method and application
  • Nitroreductase-responsive diagnostic and therapeutic integrated probe and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1. Preparation of integrated probe for diagnosis and treatment of nitroreductase response

[0046] The integrated diagnosis and treatment probe is synthesized according to the following route:

[0047]

[0048] Specific steps: put H 2 o 2 (62 μL, 0.6 mmol) and dichloromethane (2 mL) were mixed uniformly, trifluoroacetic anhydride (126 μL, 0.8 mmol) was added dropwise thereto, stirred at 0° C. for 30 min and then warmed to room temperature. Then the near-infrared fluorescent dye (41mg, 0.1mmol) was dissolved in dichloromethane (2mL) and added to the above solution to continue the reaction for 2h. The reaction solution was extracted with dichloromethane and water, and the organic phase was evaporated to dryness under reduced pressure to obtain the crude product. Chloromethane / methanol (20:1, V / V) was used as the eluent and separated by column chromatography on silica gel to obtain the integrated probe for diagnosis and treatment as a purple powder.

[0049]T...

Embodiment 2

[0050] Example 2. The optical response of the integrated diagnosis and treatment probe to nitroreductase

[0051] Weigh 4.4 mg of the integrated diagnosis and treatment probe, dissolve it in 10 mL of dimethyl sulfoxide, and prepare the probe stock solution (also known as mother solution, with a concentration of 1 mM), and make NADH stock solution with a concentration of 50 mM; mix 50 μL of the above probe The mother solution was added to a certain amount of phosphate buffered saline solution (PBS, 0.01M), then 25 μL of NADH mother solution and different amounts of nitroreductase solutions were added, and then the volume was adjusted to 5 mL with PBS. After reacting at 37°C for 10 minutes, test its ultraviolet-visible absorption spectrum and fluorescence emission spectrum. The excitation and emission slit widths were 20nm and 10nm, respectively, when the fluorescence emission spectrum was measured with excitation at 660nm.

[0052] Figure 4 Shown is the ultraviolet-visible a...

Embodiment 3

[0053] Example 3. The integrated probe for diagnosis and treatment is used for imaging of hypoxic tumor cells

[0054] (1) At 37°C and 5% CO 2 Under these conditions, human lung cancer cell A549 was cultured with DMEM medium containing 10% (v / v) fetal bovine serum (FBS), 100 U / mL penicillin, and 100 μg / mL streptomycin.

[0055] (2) Inoculate the A549 cells in the logarithmic growth phase in a laser confocal dish, discard the culture medium after culturing for 24 hours and wash with PBS (pH 7.4) three times, and then culture the cells with a medium containing 100 μM cobalt chloride to induce After hypoxia, the culture medium was removed after 12, 24, 36 and 48 hours respectively, and the cells were washed three times with PBS, and then co-incubated with the integrated diagnosis and treatment probe (10 μM) prepared by the present invention for 30 minutes, and then placed in a laser confocal microscope for imaging. The imaging excitation wavelength is 635nm, and the collection r...

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Abstract

The invention belongs to the field of biomedicine and discloses a nitroreductase-responsive integrated probe for diagnosis and treatment as well as a preparation method and application thereof. The general structural formula of the integrated diagnosis and treatment probe is as follows: the specific preparation method is as follows: first react hydrogen peroxide and trifluoroacetic anhydride in an organic solvent, then add a near-infrared fluorescent dye to continue the reaction, and after the reaction is completed, separate and purify to obtain the obtained The integrated probe for diagnosis and treatment. The integrated probe for diagnosis and treatment of the present invention can exhibit fluorescence enhancement response to nitroreductase in the tumor microenvironment, and can be used for imaging of nitroreductase and living tumors; at the same time, the integrated probe for diagnosis and treatment of the present invention can Generate reactive oxygen species for photodynamic therapy of tumors. The integrated diagnosis and treatment probe of the present invention has high biological safety, and the preparation method is simple and easy to operate, low in cost and easy to popularize, and has potential application value in the early diagnosis and treatment of malignant tumors.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a nitroreductase-responsive integrated probe for diagnosis and treatment, a preparation method and application thereof. Background technique [0002] In the past ten years, the incidence and mortality of malignant tumors have been high, and it has become one of the major public health problems that seriously threaten human health. The development of early detection methods and effective treatment methods for tumors is the key to improving the quality of life of patients and reducing deaths. rate of the only way. The integration of diagnosis and treatment is a new type of biomedical technology that combines disease diagnosis and monitoring and drug treatment. It can simultaneously have multiple functions such as tumor monitoring, anti-tumor treatment, and efficacy evaluation. Hypoxia is one of the important characteristics of the tumor microenvironment, and it is also a key...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D405/06C09K11/06A61K41/00A61K49/00A61P35/00
CPCC07D405/06C09K11/06A61K41/0057A61K49/0021A61P35/00C09K2211/1029C09K2211/1088
Inventor 李丽红张琪任国栋张敏刁海鹏刘文王浩江王斌
Owner SHANXI MEDICAL UNIV
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