Preparation method of parallel-configuration G-quadruplex DNA probe for specific detection

A technology of DNA probes and quadruplexes, applied in the field of biological probes, can solve problems such as single means and large interference, and achieve the effects of easy-to-obtain raw materials, strong operability, and low production costs

Active Publication Date: 2020-05-05
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the reported probes detect the conformation of G-quadruplex DNA in a single way, and the interference is also relatively large

Method used

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  • Preparation method of parallel-configuration G-quadruplex DNA probe for specific detection
  • Preparation method of parallel-configuration G-quadruplex DNA probe for specific detection
  • Preparation method of parallel-configuration G-quadruplex DNA probe for specific detection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: the synthesis of compound 3

[0039] Under nitrogen protection, dissolve 1.0 g (3.7 mmol) of p-hydroxybenzaldehyde (compound 1) modified with ether oxygen chains and 0.7 g (7.4 mmol) of 2,4-dimethylpyrrole in 100 mL of anhydrous dichloromethane , add 3-5 drops of catalytic amount of trifluoroacetic acid dropwise, stir at room temperature for 12 hours, then add 0.84g (3.7mmol) 2,3-dichloro-5,6-dicyano-p-benzoquinone, stir for 12 hours, then add 18mL (129mmol) of triethylamine, stirred for 30 minutes, added 15mL (118.0mmol) of boron trifluoride diethyl ether, stirred for 8 hours. Washed successively with ultrapure water (25mL×2 times) and saturated aqueous NaCl solution (25mL×2 times), the obtained organic phase was dried over anhydrous sodium sulfate, dichloromethane was distilled off under reduced pressure, and the obtained crude product was washed with ethyl acetate and petroleum The mixed solvent with ether volume ratio of 1:1 was used as the mobile pha...

Embodiment 2

[0040] Embodiment 2: the synthesis of compound 5

[0041]Dissolve 0.65g (5.0mmol) N-hydroxyethylpiperazine in 50mL of DMSO, add 0.62g (5.0mmol) 4-fluorobenzaldehyde and 2.0g (15.0mmol) K 2 CO 3 The reaction was heated at 100°C in an oil bath for 24 hours. Add 100mL of water after the reaction, and extract with dichloromethane (50mL×3 times), the organic phase obtained is dried over anhydrous sodium sulfate, dichloromethane is distilled off under reduced pressure, and the volume ratio of the resulting crude product to ethyl acetate and petroleum ether is A 2:1 mixed solvent was used as the mobile phase and silica gel was used as the stationary phase for column chromatography purification to obtain 0.80 g of yellow solid product 5 with a yield of 68.3%.

Embodiment 3

[0042] Embodiment 3: the synthesis of probe

[0043] 0.4g (0.82mmol) of compound 3, 0.19g (0.82mmol) of compound 5, 0.8mL of glacial acetic acid and 1mL of piperidine were dissolved in 50mL of toluene, and heated to 120°C for 8 hours. After the reaction, the toluene was evaporated under reduced pressure, and the obtained crude product was purified by column chromatography with a mixed solvent of dichloromethane and methanol in a volume ratio of 20:1 as the mobile phase and silica gel as the stationary phase to obtain 0.21 g of dark blue solid product probe , and the yield was 36.8%. 1 H NMR (400MHz, CDCl 3 )δ: 7.54-7.49(m, 3H), 7.19-7.15(m, 3H), 7.02(d, J=8.24Hz, 2H), 6.88(d, J=8.56Hz, 2H), 6.57(s, 1H ), 5.98(s, 1H), 4.19(t, J=4.28Hz, 2H), 3.91(t, J=4.64Hz, 2H), 3.78-3.76(m, 6H), 3.70-3.66(m, 4H) , 3.57-3.55(m, 2H), 3.38(m, 5H), 2.86(br, 4H), 2.76(m, 2H), 2.57(s, 3H), 1.46(s, 3H), 1.42(s, 3H ); 19 FNMR (376MHz, CDCl 3 )δ: -139.66, -139.73, -139.84, -139.92; HRMS (ESI) m / ...

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Abstract

The invention discloses a preparation method of parallel-configuration G-quadruplex DNA probe for specific detection, and belongs to the technical field of biological probes. The specific structural formula of the probe is shown in the specification. The probe is simple in preparation steps, easily available in raw materials and low in fluorescence background. Through the ultraviolet spectrophotometer and the fluorescence spectrophotometer, rapid bifunctional specific detection of the parallel configuration G-quadruplex DNA can be realized. The defects that a traditional detection method is single, and G-quadruplex DNA of different configurations is difficult to distinguish are overcome, and the method has wide application prospects in the fields of biochemistry, molecular biology and thelike.

Description

technical field [0001] The invention relates to the construction of a novel bifunctional probe, its preparation method and application, and belongs to the technical field of biological probes. Background technique [0002] G-quadruplex DNA is a special DNA structure, which is a guanine-rich DNA sequence that self-assembles into a G-tetrad by its own four guanine rings through Hoogsteen hydrogen bonds under certain conditions, two or more G-tetrad is an advanced DNA secondary structure formed by π-π stacking. According to the orientation of the four strands in the G-quadruplex, the G-quadruplex can be divided into three different structures, namely parallel, antiparallel and mixed structures. In recent years, bioinformatics studies have shown that there are about 370,000 groups of guanine-rich gene sequences that may form a G-quadruplex structure in the human body, especially at the ends of human telomeres and oncogene promoter regions (such as c -myc, ckit, bcl-2, Pu27, kR...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07F5/02C09K11/06G01N21/33G01N21/64
CPCC07F5/022C09K11/06G01N21/6428G01N21/33C09K2211/1044C09K2211/1007C09K2211/1055
Inventor 王明齐马瑞于全琦刘洪孛
Owner JIANGSU UNIV
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