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Small-molecule fluorescent probe used for ratio recognition of human carbonic anhydrase, and synthetic method and application thereof

A technology of carbonic anhydrase and fluorescent probes, which can be used in fluorescence/phosphorescence, chemical instruments and methods, luminescent materials, etc., and can solve urgent needs and other problems

Active Publication Date: 2017-06-20
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, ratiometric probes applied to protein recognition are rarely reported, and the demand is more urgent

Method used

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  • Small-molecule fluorescent probe used for ratio recognition of human carbonic anhydrase, and synthetic method and application thereof
  • Small-molecule fluorescent probe used for ratio recognition of human carbonic anhydrase, and synthetic method and application thereof
  • Small-molecule fluorescent probe used for ratio recognition of human carbonic anhydrase, and synthetic method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Synthesis of small molecule fluorescent probes for ratiometric recognition of human carbonic anhydrase.

[0033] (1) Synthesis of intermediate 4-(2-chloroacetyl)amino-1,8-naphthalene anhydride:

[0034] 4-Amino-1,8-naphthalene anhydride (1.0 g, 4.7 mmol) was placed in 60 mL of tetrahydrofuran, and 0.5 mL of chloroacetyl chloride was added under ice-cooling. Stir at room temperature overnight (14h), remove the solvent under reduced pressure, separate on a silica gel column with dichloromethane as eluent, and remove the solvent under reduced pressure to obtain 1.0 g of off-white solid with a yield of 76%. 1 HNMR (400MHz, DMSO) δ10.86(s, 1H), 8.80(d, J=8.8Hz, 1H), 8.59-8.63(m, 2H), 8.37(d, J=8.0Hz, 1H), 8.02( t,J=7.8Hz,1H), 4.62(s,2H).

[0035]

[0036] (2) Synthesis of intermediate 4-(2-(N,N-diethylamino)acetyl)amino-1,8-naphthalene anhydride:

[0037] 4-(2-Chloroacetyl)amino-1,8-naphthalene anhydride (500 mg, 1.7 mmol) was placed in 50 mL of acetonitrile...

Embodiment 2

[0043] Example 2: The response of the fluorescent probe prepared in Example 1 to different solvents.

[0044] Figure 5 The concentration of the fluorescent probe prepared in Example 1 is 10 μM, and the excitation light is 370nm. In 20mM pH=7.4PBS buffer solution, the probe exhibits a blue fluorescence with a wavelength of ~470nm. In methanol, dimethyl sulfoxide, N, In N-dimethylformamide, acetonitrile, and ethanol, the main fluorescence peak red-shifted to 525-560nm; indicating that the fluorescent probe can exhibit long-wave properties in a hydrophobic environment.

Embodiment 3

[0045] Example 3: The fluorescent probe prepared in Example 1 responds to 1 equivalent of carbonic anhydrase within a sustained period of time.

[0046] Image 6 The concentration of the fluorescent probe prepared in Example 1 in 20mM pH=7.4PBS buffer solution was 1 μm, and the excitation light was 405nm. After adding 1 equivalent of carbonic anhydride 1 of the fluorescent probe, the fluorescence intensity at a wavelength of ~470nm was significantly reduced, while the wavelength The fluorescence intensity at ~530nm increased sharply, and there was no significant change within 48 hours; it indicated that the fluorescent probe entered the hydrophobic cavity of carbonic anhydrase 1, and the fluorescence showed long-wave properties. .

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Abstract

The invention relates to a small-molecule fluorescent probe used for ratio recognition of human carbonic anhydrase, and a synthetic method and application thereof. The probe is prepared by reacting 4-amino-1,8-naphthalic anhydride with chloroacetyl chloride, diethylamine, 4-aminomethylbenzenesulfonamide and like. The synthetic method is simple to operate and uses cheap raw materials. The probe emits blue fluorescent light with a wavelength of ~470 nm in an aqueous solution and emits green to yellow fluorescent light with a wavelength of 525 to 560 nm in polar solvents like methanol, dimethyl sulfoxide, N,N-dimethyl formamide and ethanol. In a 20-mM phosphoric acid buffer, when human carbonic anhydrase 1 with an equivalent of 0 to 0.7 is gradually added, a fluorescence peak at a wavelength of ~470 nm gradually attenuates and a fluorescence peak at a wavelength of ~530 nm gradually intensifies. Thus, the probe provided by the invention can realize ratio recognition of human carbonic anhydrase and eliminates influence of external factors like equipment and samples.

Description

technical field [0001] The invention relates to the synthesis and application of a small molecule fluorescent probe for ratiometric identification of human carbonic anhydrase. Background technique [0002] With the rapid development of life science research, fluorescence analysis technology is more and more widely used in molecular biology, cellular immunology, oncology, medicine and other fields. Organic small molecule fluorescent probes have gradually become an important means of fluorescence analysis technology due to their advantages of small size, easy synthesis, and diversity. In the measurement of the fluorescence method, after the fluorescent probe is combined with the reactant, the excitation light or the probe whose emission spectrum wavelength shifts can be measured by using the ratio of the fluorescence intensity measured at two different wavelengths. This type of fluorescent probe is called It is a ratio probe. Compared with pure fluorescence-enhanced probes, ...

Claims

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Application Information

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IPC IPC(8): C09K11/06C07D221/14G01N21/64
Inventor 徐兆超乔庆龙
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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