Nucleic acid fluorescence probe for nuclear staining and preparation method of nucleic acid fluorescence probe

A fluorescent probe and cell nucleus technology, applied in the field of cell nucleus fluorescent probes, can solve the problems of low anti-biological metabolism and high biological toxicity, and achieve the effect of strong anti-enzymolysis ability, low cytotoxicity and high nuclear targeting

Active Publication Date: 2018-08-03
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the solution state, these fluorescent dyes usually face problems such as easy photobleaching, high biological toxicity, and low ability to resist biological metabolism.

Method used

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  • Nucleic acid fluorescence probe for nuclear staining and preparation method of nucleic acid fluorescence probe
  • Nucleic acid fluorescence probe for nuclear staining and preparation method of nucleic acid fluorescence probe
  • Nucleic acid fluorescence probe for nuclear staining and preparation method of nucleic acid fluorescence probe

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Experimental program
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Effect test

Embodiment 1

[0029] A, the preparation of intermediate 1

[0030] At room temperature, dissolve 1.26mmol 6-amino-2-methylquinoline in acetonitrile and add it to a 250ml flask, then slowly add 1.33mmol N-bromosuccinimide and a catalytic amount of silica gel, and continue stirring for 15min Afterwards, the liquid in the bottle turned dark brown, and thin-layer chromatography determined that new spots were generated in the reaction, and the raw materials were not completely reacted, and the reaction was continued for 12 hours. After the reaction was finished, extracted with ethyl acetate, spin-dried the solvent, and the residue was separated and purified through a chromatographic column (ethyl acetate:petroleum ether volume ratio=1:6) to obtain 207 mg of a yellow solid product, namely intermediate 1, with a yield of 69%.

[0031] 1 H NMR (400MHz, CDCl3, TMS): δ = 8.23 ​​(d, J = 8.6Hz, quinoline-H, 1H), 7.81 (d, J = 9.0Hz, quinoline-H, 1H), 7.28 (d, J = 8.6Hz, quinoline-H, 1H), 7.20 (d, J=9...

Embodiment 2

[0045] A. Preparation of Intermediate 1

[0046] At room temperature, 1.26mmol of 6-amino-2-methylquinoline was dissolved in acetonitrile and added to a 250ml flask, then 1.33mmol of N-bromosuccinimide and a catalytic amount of silica gel were slowly added, and continued stirring for 15min After the liquid in the bottle turned dark brown, thin layer chromatography determined that a new point was formed in the reaction, and the raw material was not completely reacted, and the reaction was continued for 16 hours. After the reaction was completed, ethyl acetate was extracted, the solvent was spin-dried, and the residue was separated and purified by a chromatographic column (ethyl acetate: petroleum ether volume ratio = 1:6) to obtain 222 mg of a yellow solid product, namely Intermediate 1, with a yield of 222 mg. was 74%.

[0047] B, the preparation of intermediate 2

[0048] Add 90 mg of intermediate 1 to the round-bottomed flask, dissolve it with 16 ml of a mixed solution of ...

Embodiment 3

[0052] A. Preparation of Intermediate 1

[0053] At room temperature, 1.26mmol of 6-amino-2-methylquinoline was dissolved in acetonitrile and added to a 250ml flask, then 1.33mmol of N-bromosuccinimide and a catalytic amount of silica gel were slowly added, and continued stirring for 15min Afterwards, the liquid in the bottle turned dark brown, and thin-layer chromatography determined that a new point was formed in the reaction, and the raw material was not completely reacted, and the reaction was continued for 18 hours. After the reaction was completed, ethyl acetate was extracted, the solvent was spin-dried, and the residue was separated and purified by a chromatographic column (ethyl acetate: petroleum ether volume ratio = 1:6) to obtain 216 mg of the product in the form of a yellow solid, namely intermediate 1, and the yield was was 72%.

[0054] B, the preparation of intermediate 2

[0055] Add 90 mg of intermediate 1 to the round-bottomed flask, dissolve it with 16 ml ...

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Abstract

The invention discloses a preparation method of a nucleic acid fluorescence probe compound AzosD for nuclear staining. The preparation method is characterized by comprising steps as follows: 6-amino-2-methylquinoline is dissolved in acetonitrile, N-bromosuccinimide is added, a reaction is performed, and an intermediate 1 is obtained; the intermediate 1 is dissolved in a water, ethanol and toluenemixed solution, 3-(N,N-dimethylamino)phenylboronic acid, tetrakis(triphenylphosphine)palladium and sodium carbonate are added, refluxing reaction is performed under nitrogen protection, and an intermediate 2 is obtained; the intermediate 2 is dissolved by hydrochloric acid, sodium nitrite is added to the solution under the condition of ice-water bath, pH of the solution is regulated to be neutralby a sodium hydroxide aqueous solution, and 3-methyl-11-(N,N-dimethylamino) cinnoline[3,4-e]quinoline, namely AzosD, is obtained by separation and purification. The compound is good in photobleachingresistant effect, high in biocompatibility, high in enzyme-resistant capacity and high in nuclear targeting, can be applied to specific staining of nucleuses of living cells, and has broad applicationprospect in the fields of long-term monitoring of nuclear morphometry and observation for nuclear morphology change in the cellular physiological process.

Description

technical field [0001] The invention belongs to the technical field of cell nucleus fluorescent probes, in particular to a nucleic acid fluorescent probe 3-methyl-11-(N,N-dimethylamino)cinnolino[3,4-e]quinone used for cell nucleus staining Phenyl (AzosD) and its preparation method. Background technique [0002] With the development of science and technology, human beings have a deeper understanding of the nucleus. The nucleus is the largest and most important organelle in eukaryotic cells, and it is the regulatory center of cell inheritance and metabolism. The nucleus is the control center of the cell, plays an important role in the metabolism, growth and differentiation of the cell, and is the main location of genetic material. Nucleic acid is a biological macromolecule closely related to life activities and diseases. Therefore, the staining, tracking and morphological observation of the nucleus are of great significance for understanding the life process of the whole ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D471/04C09K11/06G01N21/64
CPCC07D471/04C09K11/06C09K2211/1029C09K2211/1044G01N21/6428
Inventor 宋钦华李琛
Owner UNIV OF SCI & TECH OF CHINA
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