P-dimethylaminostyrylquinoline derivative, synthesis method thereof and application thereof in nucleic acid fluorescent probe

Abstract

The invention belongs to the technical field of biology, and discloses a P-dimethylaminostyrylquinoline derivative, a synthesis method thereof and application thereof in a nucleic acid fluorescent probe. The derivative has a chemical structural formula shown as a formula 1, R is a group disclosed in the invention or has a large electron conjugate system plane for dimethylaminostyrylquinoline derivatives, the charge transfer effect in probe molecules can influence the fluorescence emission intensity of the molecules, and G-quadruplex DNA can be specifically recognized in an in-vitro test; the fluorescent probe provided by the invention has the advantages of high fluorescence intensity, strong binding constant, low detection limit and the like.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and particularly relates to p-dimethylaminostyrene quinoline derivatives, their synthesis method and their application in nucleic acid fluorescent probes. Background technique

[0002] As an important biological macromolecule, nucleic acid undertakes various important biological functions in living cells. Common nucleic acids include RNA, double-stranded DNA, etc. In addition, there are special nucleic acid structures that have attracted extensive research in recent years. - quadruplex. RNA can encode, transcribe and control gene expression in living cells, and regulate various life activities. G-quadruplex DNA is mainly located in important sites such as gene telomeres and promoters, and plays an important role in biological switches. Therefore, it is of great significance to develop nucleic acid fluorescent probes with excellent performance for the study of the morphology and function of nucleic ...

Examples

Embodiment 1

[0042] Embodiment 1: the synthesis of intermediate a

[0043] 4-Chloro-2-methylquinoline (1.5 mmol) and iodomethane (4.5 mmol) were added to a closed pressure reactor, and then the compound was dissolved with sulfolane (6 mL). The mixture was heated and reacted at 60°C for 12 hours. After the reaction, the mixture was cooled to room temperature, and ethyl acetate (15 mL) was added to precipitate the crude product. Recrystallized in ethanol and dried to obtain dark green pure product.

[0044] The reaction process is shown in the following formula:

[0045]

Embodiment 2

[0046] Embodiment 2: the synthesis of compound B1

[0047]Intermediate a (0.2 mmol), morpholine (0.6 mmol) and 4-dimethylaminobenzaldehyde (0.4 mmol) were added to the flask. The compound was dissolved with n-butanol (4 mL). The mixture was stirred and refluxed at 80 °C for 8 h. When the reaction was complete, the mixture was cooled to room temperature, then ethyl acetate (10 mL) was added. After standing still for 30 min, the crude product was obtained by suction filtration. The target compound was obtained by recrystallization from ethanol. After drying, 0.0686 g of dark green crystals was obtained. Thin plate chromatography showed no by-products, and the crude yield was 67%. Proton spectrum data: 1 H NMR (400MHz, DMSO-d6) δ8.73 (d, J=8.3Hz, 1H), 8.09–7.97 (m, 3H), 7.86–7.79 (m, 3H), 7.77 (s, 1H), 7.75 ( d,J=7.3Hz,1H),6.82(d,J=8.5Hz,2H),4.07(s,3H),3.91–3.86(m,4H),3.69(t,J=4.5Hz,4H), 3.05(s,6H). See attached for mass spectrometry data figure 1 . The reaction proces...

Embodiment 3

[0049] Embodiment 3: the synthesis of compound B2

[0050] Intermediate a (0.2 mmol), pyrrolidine (0.6 mmol) and 4-dimethylaminobenzaldehyde (0.4 mmol) were added to the flask. The compound was dissolved with n-butanol (4 mL). The mixture was stirred and refluxed at 80 °C for 8 h. When the reaction was complete, the mixture was cooled to room temperature, then ethyl acetate (10 mL) was added. After standing still for 30 min, the crude product was obtained by suction filtration. The target compound was obtained by recrystallization from ethanol. Drying gave 0.0701 g of orange crystals, thin plate chromatography showed no by-products, and the crude yield was 70%. The hydrogen spectra data are 1 H NMR (400MHz, DMSO-d6) δ8.55 (d, J = 7.9Hz, 1H), 7.95–7.93 (m, 1H), 7.93–7.91 (m, 1H), 7.83 (d, J = 15.8Hz, 1H),7.77–7.74(m,2H),7.71(d,J=15.9Hz,1H),7.63(ddd,J=8.2,5.2,2.9Hz,1H),7.59(s,1H),6.82–6.78 (m, 2H), 3.99 (s, 3H), 3.94–3.88 (m, 4H), 3.03 (s, 6H), 2.03 (p, J=3.4Hz, 4H). See...

technical field

[0001] The invention belongs to the field of biotechnology, and particularly relates to p-dimethylaminostyrene quinoline derivatives, their synthesis method and their application in nucleic acid fluorescent probes. Background technique

[0002] As an important biological macromolecule, nucleic acid undertakes various important biological functions in living cells. Common nucleic acids include RNA, double-stranded DNA, etc. In addition, there are special nucleic acid structures that have attracted extensive research in recent years. - quadruplex. RNA can encode, transcribe and control gene expression in living cells, and regulate various life activities. G-quadruplex DNA is mainly located in important sites such as gene telomeres and promoters, and plays an important role in biological switches. Therefore, it is of great significance to develop nucleic acid fluorescent probes with excellent performance for the study of the morphology and function of nucleic ...

Examples

Embodiment 1

[0042] Embodiment 1: the synthesis of intermediate a

[0043] 4-Chloro-2-methylquinoline (1.5 mmol) and iodomethane (4.5 mmol) were added to a closed pressure reactor, and then the compound was dissolved with sulfolane (6 mL). The mixture was heated and reacted at 60°C for 12 hours. After the reaction, the mixture was cooled to room temperature, and ethyl acetate (15 mL) was added to precipitate the crude product. Recrystallized in ethanol and dried to obtain dark green pure product.

[0044] The reaction process is shown in the following formula:

[0045]

Embodiment 2

[0046] Embodiment 2: the synthesis of compound B1

[0047]Intermediate a (0.2 mmol), morpholine (0.6 mmol) and 4-dimethylaminobenzaldehyde (0.4 mmol) were added to the flask. The compound was dissolved with n-butanol (4 mL). The mixture was stirred and refluxed at 80 °C for 8 h. When the reaction was complete, the mixture was cooled to room temperature, then ethyl acetate (10 mL) was added. After standing still for 30 min, the crude product was obtained by suction filtration. The target compound was obtained by recrystallization from ethanol. After drying, 0.0686 g of dark green crystals was obtained. Thin plate chromatography showed no by-products, and the crude yield was 67%. Proton spectrum data: 1 H NMR (400MHz, DMSO-d6) δ8.73 (d, J=8.3Hz, 1H), 8.09–7.97 (m, 3H), 7.86–7.79 (m, 3H), 7.77 (s, 1H), 7.75 ( d,J=7.3Hz,1H),6.82(d,J=8.5Hz,2H),4.07(s,3H),3.91–3.86(m,4H),3.69(t,J=4.5Hz,4H), 3.05(s,6H). See attached for mass spectrometry data figure 1 . The reaction proces...

Embodiment 3

[0049] Embodiment 3: the synthesis of compound B2

[0050] Intermediate a (0.2 mmol), pyrrolidine (0.6 mmol) and 4-dimethylaminobenzaldehyde (0.4 mmol) were added to the flask. The compound was dissolved with n-butanol (4 mL). The mixture was stirred and refluxed at 80 °C for 8 h. When the reaction was complete, the mixture was cooled to room temperature, then ethyl acetate (10 mL) was added. After standing still for 30 min, the crude product was obtained by suction filtration. The target compound was obtained by recrystallization from ethanol. Drying gave 0.0701 g of orange crystals, thin plate chromatography showed no by-products, and the crude yield was 70%. The hydrogen spectra data are 1 H NMR (400MHz, DMSO-d6) δ8.55 (d, J = 7.9Hz, 1H), 7.95–7.93 (m, 1H), 7.93–7.91 (m, 1H), 7.83 (d, J = 15.8Hz, 1H),7.77–7.74(m,2H),7.71(d,J=15.9Hz,1H),7.63(ddd,J=8.2,5.2,2.9Hz,1H),7.59(s,1H),6.82–6.78 (m, 2H), 3.99 (s, 3H), 3.94–3.88 (m, 4H), 3.03 (s, 6H), 2.03 (p, J=3.4Hz, 4H). See...

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