Bis-benzimidazole fluorescent dye as well as preparation method and application thereof
A bibenzimidazole and fluorescent dye technology, applied in the field of biological fluorescent dyes, can solve the problems of increased post-processing difficulty, a large amount of impurities, and reduced yields, and achieve rapid labeling and detection of DNA, mild conditions, and improved sensitivity and accuracy Effect
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Embodiment 1
[0040] Preparation of compound 3
[0041] Combine 5-chloro-2-nitroaniline (2.59g, 15mmol), methylpiperazine (1.5g, 15mmol) and carbonic acid
[0042] Potassium (2.1 g, 15 mmol) was added to DMF, mixed, and refluxed until the reaction was complete. The mixture was poured into cold water, extracted with ethyl acetate, the organic layer was separated, washed with water, dried over anhydrous sodium sulfate, concentrated, and subjected to silica gel column chromatography to obtain 3 g of a pale yellow solid with a yield of 85%. 1 H NMR (500 MHz, DMSO) δ7.83 (d, 1H), 7.13 (s, 1H), 6.39 (d, 1H), 6.27 (s, 1H), 3.61 – 3.53 (t, 2H), 3.47 – 3.39 (t, 2H), 2.55 (t, 2H), 2.40 (t, 2H), 2.25 (s, 3H).
Embodiment 2
[0044] Preparation of compound 4
[0045] Compound 3 (30 g, 127 mmol) and 5% palladium on carbon (18 g) were added to anhydrous ethanol, and at room temperature, hydrogen
[0046] Hydrogenation was carried out in the solution, the catalyst was removed by filtration, and the solvent was evaporated to dryness to obtain 24.5 g of a yellow-brown precipitate with a yield of 93.5%. 1 H NMR (500 MHz, CDCl 3 ) δ = 6.26 (d, 1H), 6.02 (m, 1H), 5.78 (d, 1H), 4.22 (m, 4H), 3.69 – 3.59 (t, 2H), 3.53 – 3.43 (t, 2H), 2.66 – 2.54 (t, 2H), 2.47 –2.38 (t, 2H), 2.29 (s, 3H).
Embodiment 3
[0048] Preparation of compound 5a
[0049] 3-Nitro-4-aminobenzonitrile (4.8 g, 30 mmol) was dissolved in absolute ethanol. After cooling in the ice bath, hydrogen chloride gas was bubbled through the solution, maintaining the temperature between 0-1 ºC. The reaction mixture was stored in a refrigerator (4°C) for 4-5 days to obtain an orange-yellow precipitate, which was filtered and washed with anhydrous ether to obtain a precipitate of 7.2 g with a yield of 98%. 1 H NMR (500 MHz, DMSO) δ 7.54 (s, 1H), 7.44 (dd, 1H), 6.94 (d, 1H), 3.67 (m, 2H), 1.31 (t, 3H).
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